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Preparation and sensitization of polystyrene latex beads by some antigens and antibodies. Factors affecting sensitivity and specificity of latex agglutination tests.

Polystyrene latex beads were polymerized at two different pH (7.3 and 9.5) and their ability to create latex agglutination systems were studied. Sensitization of these latexes by DNA, antibodies to C-reactive protein, myoglobin, and human IgG. was conducted. Sensitivity and specificity of these systems were compared. Practical recommendations are given for preparation of such systems.

Animals↗

Fission yeast sts1+ gene encodes a protein similar to the chicken lamin B receptor and is implicated in pleiotropic drug-sensitivity, divalent cation-sensitivity, and osmoregulation.

The Schizosaccharomyces pombe sts1+ gene, identified by supersensitive mutations to a protein kinase inhibitor, staurosporine, was isolated by complementation by the use of a fission yeast genomic library. Nucleotide sequencing shows that the sts1+ gene encodes a 453 amino acid putative membrane-associated protein that is significantly similar (26% identity) to the chicken lamin B receptor. It is also highly related (53% identity) to a budding yeast ORF, YGL022. These three proteins contain a similar hydrophobicity pattern consisting of eight or nine putative transmembrane domains. By gene disruption we demonstrate that the sts1+ gene is not essential for viability. These disruptants exhibit pleiotropic defects, such as cold-sensitivity for growth and at the permissive temperature, a supersensitivity to divalent cations and several unrelated drugs including staurosporine, caffeine, chloramphenicol, sorbitol, and SDS. Disruption of the sts1+ gene does not lead to a sensitivity to thiabendazole or hydroxyurea.

Amino Acid Sequence↗

Contrast sensitivity in 24- and 36-month-olds as assessed with the contrast sensitivity card procedure.

We used a new card procedure to evaluate binocular contrast sensitivity (CS) in 24- and 36-month-old children. The test consists of 40 large (50 by 28 cm) matteboard cards, each of which contains a sine wave grafting with 1 of 5 spatial frequencies (0.4, 0.8, 1.6, 3.2, and 4.8 cpd at 80 cm) and with 1 of 8 contrast levels [from 33% (CS = 3) to 0.4% (CS = 260)]. Estimates of subjects' CS to each spatial frequency were obtained with a modified forced-choice preferential looking (FPL) technique similar to that developed for the Teller Acuity Cards (TAC). All 36-month-olds and 74% of the 24-month-olds completed the entire procedure in an average of 12 min per child. For both groups, the shape of the mean constant sensitivity functions (CSF's) resembled the characteristic inverted U shown by normal adults, except that the toddlers' functions were shifted to lower contrasts and spatial frequencies. Combined with previous results, it appears that the CS cards hold promise as a time-efficient means of assessing CS in infants and young children, and may provide the prototype for a clinical tool to help detect early visual and neurological dysfunction.

Child, Preschool↗

Brief reactive psychosis induced by sensitivity training: similarities between sensitivity training and brainwashing situations.

Sensitivity training (ST), which originated in the USA during the late 1940s, has been used as part of training seminars in Japanese corporations since the late 1950s. The possibility of negative psychiatric effects of ST, and especially its role in inducing psychiatric symptoms, is yet to be clarified. A case of a 41-year-old male company worker whose brief psychosis was induced by a sensitivity training seminar held by the company he worked for is presented. In reviewing the psychopathology of the case with records from the ST seminar, we found similarities between the patient's ST seminar and brainwashing situations. Specifically, the patient experienced severe conflict (of thought process) between his Christian beliefs and being labeled a coward at the seminar. We conclude that monitoring of the ST programs is crucial in order to ensure the psychological safety of ST participants in Japan.

Adult↗

Information derived from sensitization test methods: test sensitivity, false positives and false negatives.

Predictive toxicology tests for the prospective identification of skin-sensitizing chemicals are well known and have been used for many years. However, of these, only the local lymph node assay (LLNA) has actually undergone formal independent assessment to determine the accuracy of the predictions, particularly with respect to the likelihood of false positives and false negatives. Often, efforts to increase the sensitivity of a test (reducing false negatives) tend to increase the number of false positives. In this short review, these issues are discussed in particular relation to the 3 predictive tests available in regulatory toxicology, the guinea-pig maximization test, the occluded patch test of Buehler and the LLNA. A key perspective is that no predictive test is without limitations; having a good appreciation of these limitations is necessary for making the best use of the information derived from these methods.

Animals↗

Sensitivity and specificity of an improved rapid latex agglutination test for identification of methicillin-sensitive and -resistant Staphylococcus aureus isolates.

The performance of a second-generation rapid agglutination kit, Slidex Staph Plus (SSP; bioMérieux), was compared to those of the Slidex Staph (SS; bioMérieux), Staphaurex (SRX; Murex Diagnostics), and BBL Staphyloslide (BBL; Becton Dickinson) kits by using 508 clinical isolates composed of 150 methicillin-sensitive Staphylococcus aureus (MSSA) organisms, 154 methicillin-resistant S. aureus (MRSA) organisms, and 204 non-S. aureus Staphylococcus spp. Of the 508 isolates tested, 75% were fresh clinical isolates, with the remainder taken from five different freezer collections. All four agglutination tests had comparable sensitivities for MSSA and MRSA. However, the SS kit was significantly less specific (93.1%) than the three other tests (P > 0.05, McNemar test). These results demonstrate that the new rapid latex agglutination kit, SSP, was more specific for the identification of S. aureus than the previous version and performed comparably to the SRX and BBL kits.

DNA, Bacterial↗

Assessing anxiety sensitivity in inner-city African American children: psychometric properties of the childhood anxiety sensitivity index.

This study examined the psychometric properties of the Childhood Anxiety Sensitivity Index (CASI; Silverman et al., 1991) in a sample of urban African American elementary school children. One hundred forty-four 4th- and 5th-grade children completed the CASI as part of a larger project. In contrast to prior research with community samples, CASI total scores were higher and means did not differ by sex. Internal consistency (alpha =.82), 2-week test-retest reliability (r =.80), and convergent and divergent validity were good and consistent with prior research. The factor structure of the CASI differed from the structure found with White children. Results of exploratory factor analyses suggest two factors: physical concerns and mental incapacitation concerns. Implications for assessing anxiety and anxiety sensitivity among African American children are discussed.

Black or African American↗

Monitoring thyroglobulin in a sensitive immunoassay has comparable sensitivity to recombinant human tsh-stimulated thyroglobulin in follow-up of thyroid cancer patients.

CONTEXT: Most thyroglobulin (Tg) assays have a sensitivity of 0.5-1 ng/ml. A minority of patients with undetectable T4-suppressed Tg levels have a recombinant human TSH (rhTSH)-stimulated Tg above 2 ng/ml and identifiable residual disease. OBJECTIVE: The objective was to determine whether a Tg assay with improved sensitivity could eliminate the need for rhTSH stimulation when baseline Tg is below 0.1 ng/ml. DESIGN: A retrospective study of two academic endocrine practices was conducted. POPULATION: A total of 194 patients undergoing rhTSH stimulation participated in the study. RESULTS: Of the 80 patients with Tg below 0.1 ng/ml, two (2.5%) had rhTSH-stimulated Tg above 2 ng/ml. One other patient with stimulation to 0.3 ng/ml and negative 123I scan had an ultrasound-detected malignant lymph node resected. None had 131I/123I imaging after rhTSH stimulation suggestive of local recurrence or distant metastasis. If T4-suppressed Tg was 0.1-0.5 or 0.6-2.0 ng/ml, rhTSH Tg was above 2 ng/ml in 24.2 and 82.4%, respectively. CONCLUSIONS: Patients with differentiated thyroid carcinoma and a T4-suppressed serum Tg below 0.1 ng/ml rarely have a rhTSH-stimulated Tg above 2 ng/ml, and none of these patients had 131I or 123I imaging after rhTSH stimulation suggestive of local recurrence or distant metastasis. We recommend monitoring such patients with a T4-suppressed Tg level and periodic neck ultrasonography. An increase in T4-suppressed serum Tg to a detectable level or the appearance of abnormal lymph nodes by physical or ultrasound exam should prompt further investigation.

Adolescent↗

[Report on the 89th Scientific Assembly and Annual Meeting of the Radiological Society of North America--balance between the time sensitivity profile and the slice sensitivity profile in multi-slice helical scanning].

PURPOSE: With multi-slice helical scanning, increasing helical pitch causes changes in the effective slice sensitivity profile and thus in a thickening of the effective slice thickness. However, the increase in helical pitch alters the time sensitivity profile, thereby decreasing the effective scan time. In this paper we report the results of our research concerning the setting of optimum conditions to resolve this issue. METHOD AND MATERIALS: The experiment was conducted with a moving phantom. The movement showed a variety of changes. RESULTS: When the subject was not in motion, increases in helical pitch resulted in deterioration in the image quality of the CT image. However, when the subject was in motion increases in helical pitch improved image quality. CONCLUSION: The general belief until now has been that helical pitch should be decreased in order to improve CT image quality. However, in the case of a moving subject, reduced helical pitch leads to deterioration in the time resolution and thus to a deterioration in the image quality. Through this study we have acquired an understanding of which helical pitch value is optimal in terms of the subject's rate of movement.

Sensitivity and Specificity↗

Effects of sensitizers on cell respiration: II. The effects of hypoxic cell sensitizers on oxygen utilization in cellular and chemical models.

The metabolic activity of nitroheterocyclic sensitizers could limit their usefulness in vivo. Biochemical mechanisms of drug metabolism, toxicity and effects on cell respiration have been studied in microsomes, and the kinetics of the simulated redox reactions determined by pulse radiolysis. Stimulated oxidation of coenzyme, glucose, ascorbate or glutathione substrate radicals by nitroheterocyclic sensitizers, with the concomitant appearance of the respective nitro radical anions, is observed. Under hypoxia, the nitro radical anions decay slowly by second order processes, forming reduced metabolites. In air, the nitro radical anions react with oxygen forming superoxide radical anions, peroxide and regenerating the drug. Nitro radical-anions also react with cytochrome-c indicating a possible interference with mitochondrial energy metabolism.

Animals↗

[Development of a method for the quantitative assessment of microorganism sensitivity to antibiotics using discs. The study of different nutrient media for determining microorganism sensitivity to antibiotics].

Five nutrient media used for determination of microbial sensitivity to antibiotics, i.e. beaf-peptone agar, Hottinger pancreatic beaf infusion agar, sprat hydrolysate nutrient agar of the Dagestan Research Institute of Nutrient Media, Muller-Chintone agar from Bulgaria and "Oxoid" agar for determination of microbial sensitivity were studied comparatively. The media were compared with respect to the growth density with the use of different test-cultures and the clearance of the inhibition growth zones around the discs containing different antibiotics. The best results were obtained with the use of sprat hydrolysate nutrient agar. Further studies on the medium standardization are necessary.

Anti-Bacterial Agents↗

[Standardization of the methods of determining microorganism sensitivity to antibiotics. The importance of the method of interpreting the results in determining microorganism sensitivity to antibiotics].

The significance of the methods of the data interpretation for determination of the microbial sensitivity to antibiotics is discussed. A necessity for using semi-quantitative or quantitative systems of calculation instead of qualitative interpretation of the inhibition growth zone diameters is presented. It is shown that the use of the standard nutrient medium for determination of the antibiotic sensitivity in the laboratory practice may be a base for applying the methods of interpretation of the diffusion test data.

Agar↗

Sesquiterpene lactone dermatitis. Cross-sensitivity in costus-sensitized patients.

Thirteen costus-sensitive patients were patch tested with 38 sesquiterpene lactones of five different classes over a two-year period. Cross-reacting agents fell into two chemical categories: (1) those that resembled the primary sensitizer, and (2) those belonging to different skeletal classes. An exocyclic methylene group conjugated to a gamma-lactone was present in both chemicals that cross-reacted and those that did not. The difference between these two groups is that cross-reacting chemicals are not highly substituted, tending to be lipophilic, while those giving negative responses all are highly substituted at the C-8/C-6 position. This functional group may hinder binding of exocyclic methylene with skin protein or the actual antigenic site with an immune receptor cell.

Cross Reactions↗

Naloxone-sensitive, haloperidol-sensitive, [3H](+)SKF-10047-binding protein partially purified from rat liver and rat brain membranes: an opioid/sigma receptor?

A naloxone-sensitive, haloperidol-sensitive, [3H](+)SKF-10047-binding protein was partially purified from rat liver and rat brain membranes in an affinity chromatography originally designed to purify sigma receptors. Detergent-solubilized extracts from membranes were adsorbed to Sephadex G-25 resin containing an affinity ligand for sigma receptors: N-(2- 3,4-dichlorophenyl]ethyl)-N-(6-aminohexyl)-(2-[1-pyrrolidinyl]) ethylamine (DAPE). After eluting the resin with haloperidol, a protein that bound [3H](+)SKF-10047 was detected in the eluates. However, the protein was not the sigma receptor. [3H](+)SKF-10047 binding to the protein was inhibited by the following compounds in the order of decreasing potency: (+)pentazocine > (-) pentazocine > (+/-)cyclazocine > (-)morphine > (-)naloxone > haloperidol > (+)SKF-10047 > DADLE > (-)SKF-10047. Further, the prototypic sigma receptor ligands, such as 1,3-di-o-tolylguanidine (DTG), (+)3-PPP, and progesterone, bound poorly to the protein. Tryptic digestion and heat treatment of the affinity-purified protein abolished radioligand binding. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE) of the partially-purified protein from the liver revealed a major diffuse band with a molecular mass of 31 kDa, a polypeptide of 65 kDa, and another polypeptide of > 97 kDa. This study demonstrates the existence of a novel protein in the rat liver and rat brain which binds opioids, benzomorphans, and haloperidol with namomolar affinity. The protein resembles the opioid/sigma receptor originally proposed by Martin et al. [(1976): J. Pharmacol. Exp. Ther., 197:517-532.]. A high degree of purification of this protein has been achieved in the present study.

Animals↗

Correlations between structure, topographic arrangement, and spectral sensitivity of sound-sensitive interneurons in crickets.

The morphology of nine prothoracic, sound-activated, interganglionic interneurons in Teleogryllus oceanicus is described. Only two of the neurons can, on anatomical grounds, receive input directly from auditory receptors. The morphology of many of the cells suggests that they may provide output to motor areas. The nine cells can be divided into two groups on the basis of their spectral sensitivity: high-frequency neurons and low-frequency neurons. Correlations were found between morphology and spectral sensitivity. High-frequency neurons have a ventromedial soma, dorsally positioned neuropile processes, and an axon in the lateral half of the promesothoracic connective. In contrast, low-frequency neurons have a dorsal and/or laterally positioned soma, neuropile processes in the ventral portion of the prothoracic ganglion, and an axon projecting in the medial half of the connective. These findings reveal the existence of a crude tonotopic organization of central neurons. In addition, they provide hints as to the type of output and the targets of these neurons.

Animals↗

Syngeneic sensitization of mouse lymphocytes on monolayers of thyroid epithelial cells. III. Induction of thyroiditis by thyroid-sensitized T lymphoblasts.

During in vitro sensitization of CBA spleen lymphocytes on syngeneic monolayers of either thyroid epithelial cells (TEC) or fibroblasts, stimulation occurs, as assessed by thymidine incorporation and lymphoblast generation. When lymphoblasts generated on syngeneic TEC are injected either into thyroid lobes of intact CBA recipients or i.v., thyroiditis appears at least 21 days after injection. Thyroiditis is assessed by both optical and electron microscopic blind studies and by the presence of antibodies in the sera directed against mouse thyroglobulin. In contrast, control animals which received identical numbers of lymphoblasts generated on syngeneic fibroblasts behaved as normal. Thyroiditis was also induced using pure thyroid-sensitized T lymphoblasts. This is the first report showing the role of T lymphocytes in the induction of experimental autoimmune thyroiditis.

Animals↗

Drug-resistant human bladder-cancer cells are more sensitive to adenovirus-mediated wild-type p53 gene therapy compared to drug-sensitive cells.

We investigated the therapeutic potential and molecular mechanism of adenovirus-mediated wt p53 gene therapy for drug-resistant human bladder cancers. KK47, a human bladder-cancer cell line, along with the drug-resistant sublines KK47/DDP10, KK47/DDP20 (cisplatin-resistant) and KK47/ADM (doxorubicin-resistant) were used for the experiments. All 4 KK47 cell lines had genetically normal p53 genes. Using an in vitro cytotoxicity assay, the drug-resistant cell lines were more sensitive to Ad-CMV-p53 cell killing than the KK47 parental cell line. Ad-CMV-p53 induced higher levels of p53 protein and mRNA in the drug-resistant cell lines than in the parental cell line and, consequently, higher levels of p21 and Bax mRNA, which resulted in higher percentages of G(1) cell-cycle arrest and apoptosis. The higher efficiencies of adenoviral gene transfer in the drug-resistant cell lines were confirmed by X-gal staining after infection with Ad-CMV-beta-gal. In conclusion, adenovirus-mediated wt p53 gene therapy was more effective in the drug-resistant bladder-cancer cell lines than in the drug-sensitive bladder-cancer cell line.

Adenoviridae↗

Modulation of cis-diamminedichloroplatinum(II) accumulation and sensitivity by forskolin and 3-isobutyl-1-methylxanthine in sensitive and resistant human ovarian carcinoma cells.

We have determined the effect of forskolin, an adenyl cyclase agonist, and 3-isobutyl-1-methylxanthine (IBMX), a phosphodiesterase inhibitor, on the accumulation and cytotoxicity of cisplatin (DDP) in 2008 human ovarian carcinoma cells. In DDP-sensitive 2008 cells, forskolin and IBMX caused 2.1-fold and 2.3-fold increases, respectively, in the short-term accumulation of DDP relative to untreated cells. The inactive analogue, 1,9-dideoxyforskolin, decreased DDP accumulation. Forskolin and IBMX also increased accumulation in A2780 cells. Neither forskolin nor IBMX had any effect on DDP accumulation in DDP-resistant 2008 cells. The effects were detectable as early as 1 min and persisted at 60 min. The concentrations for half-maximal stimulation of DDP accumulation were approximately 0.2 microM for forskolin and 0.2 mM for IBMX. Forskolin caused marked increases in cAMP levels in both sensitive and resistant 2008 cells within 1 min, although there were differences in the subsequent time-courses of the response. Both 2008 cell types had identical cAMP-dependent protein kinase (PKA) activity. These results suggest that there is a target downstream of PKA that is an important participant in DDP accumulation, and that this target is defective or missing in DDP-resistant cells. Following a 1-hr exposure to drugs, forskolin and IBMX at concentrations that were by themselves completely non-toxic increased the slopes of the clonogenic survival vs. DDP concentration curves in 2008 cells 1.9-fold and 3.3-fold, respectively. In DDP-resistant 2008 cells, however, forskolin and IBMX increased the slopes only 1.2 and 2.6-fold, respectively. These effects of forskolin and IBMX on DDP cytotoxicity did not directly correlate with the effects on the 1-hr DDP accumulation which suggested that, in addition to modulating DDP accumulation, these agents increase the cytotoxicity of the intracellular platinum. The results indicate that modulation of cAMP levels can have important effects on DDP accumulation and cytotoxicity in 2008 cells and that these effects are significantly diminished in DDP-resistant cells.

1-Methyl-3-isobutylxanthine↗