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Antigen receptors on murine T lymphocytes in contact sensitivity. II. Presentation and characterization of syngeneic anti-idiotype serum against DNFB-sensitized T cells.

This manuscript describes the preparation and characterization of a syngeneic antiserum raised in BALB/c mice against nylon wool-purified lymph node (LN) T cells from 2,4-dinitrofluorobenzene- (DNFB) sensitized BALB/c mice. This antiserum (anti-T DH-DNP) plus complement blocks the ability of LN or purified T cells from DNFB-sensitized donors to transfer immunity to naive recipients. The inhibitory activity of the serum is due to antibodies; these antibodies have specificity for idiotype (Id) determinants as shown by their ability to bind to purified mouse anti-DNP antibodies but not to rabbit anti-DNP or normal mouse Ig. Inhibition of transfer of immunity by the anti-Id serum is not restricted by either H-2 gene products or Igh-1 allotypes. The antiserum is antigen specific when tested on LN cells from oxazolone-sensitized mice, but has a low level of cross-reactivity against T cells from 2,4,6-trinitrochlorobenzene- (TNCB) sensitized mice. In addition, it was shown that naturally occurring anti-Id antibodies, epsilon-DNP-L-lysine, or DNP-protein (but not NIP-protein) and the anti-Id in syngeneic anti-T DH-DNP serum compete for similar binding sites on DNFB-immune T cells. We interpret these findings to indicate that immunization of mice with T cells from DNFB-sensitized syngeneic mice induces the production of anti-Id antibodies. These anti-Id antibodies are specific for determinants expressed on the T cell receptor or receptor subsite that is specific for the hapten DNP.

Absorption↗

Sensitivity of passive cutaneous anaphylaxis in rats. I. Inverse relationship between PCA sensitivity and amount of IgE present on mast cells.

Passive cutaneous anaphylaxis (PCA) sensitivity and IgE antibody production were examined in rats from four different outbred colonies: Wistar A, Wistar B, Sprague-Dawley, and Donryu. PCA sensitivity was the highest in Wistar B and the lowest in Wistar A rats using anti-Ascaris, antibovine serum albumin, and anti-Clonorchis sinensis IgE antibodies. The PCA sensitivity related inversely to the reverse PCA reaction with rabbit antirat IgE antibody, suggesting that preexisting IgE molecules on the mast cells in the skin interfered with the PCA reaction. However, the skin sensitivity to compound 48/80 was comparable in rats from the four colonies indicating the same sensitivity to nonimmunological stimulation. Ability of rats to produce IgE antibody was tested by immunizing them with dinitrophenol-coupled Ascaris extract and Bordetella pertussis adjuvant. Essentially no difference in antidinitrophenol-coupled Ascaris extract IgE antibody production was observed among animals from the different colonies.

Animals↗

Blood pressure sensitivity to salt, calcium metabolism and insulin sensitivity in essential hypertension.

1. To reveal the mechanism responsible for the increase in blood pressure (BP) response to high salt intake in salt-sensitive essential hypertensives, pressor responses to angiotensin II and norepinephrine on high and low salt diet, influence of high salt intake on calcium metabolism and insulin sensitivity were studied in essential hypertensive and in normotensive subjects with or without family history of hypertension. 2. Pressor responses to angiotensin II and norepinephrine were significantly increased in salt-sensitive essential hypertensives irrespective of high or low salt diet. 3. In normotensive subjects with positive family history, high salt diet elevated BP in association with an increase in urinary calcium excretion. This observation, however, was not shared by the normotensive subjects with negative family history of hypertension. In the normotensive subjects with positive family history of hypertension, the cytosolic calcium concentration in platelets was significantly higher than in those with negative family history. A significant correlation between the cytosolic calcium concentration and elevation of BP was also observed. 4. In normotensive subjects with family history of hypertension, insulin sensitivity was decreased and correlated negatively with the cytosolic calcium concentration. 5. These results suggest that in salt-sensitive essential hypertensives increased pressor responses to angiotensin and norepinephrine and elevation of BP as well as insulin resistance may result from inherited abnormalities of intracellular calcium metabolism.

Aged↗

Behavioral sensitivity and ethanol potentiation of the N-methyl-D-aspartate receptor antagonist MK-801 in a rat line selected for high ethanol sensitivity.

The role of the N-methyl-D-aspartate (NMDA) receptors in differential ethanol sensitivity of the alcohol-insensitive [alcohol-tolerant (AT)] and alcohol-sensitive [alcohol-nontolerant (ANT)] rat lines selected for low and high sensitivity to ethanol-induced (2 g/kg) motor impairment was studied in behavioral and neurochemical experiments. A noncompetitive antagonist of the NMDA receptor, dizocilpine maleate (MK-801; 0.2 mg/kg), impaired motor function in ANT rats, but not in AT rats, in a tilting plane test. The impairment was further potentiated by a dose (0.75 g/kg) of ethanol, which alone was inactive. This effect was apparently not associated with the locomotor stimulation produced by MK-801 (0.1 and 0.2 mg/kg), because stimulation did not differ between the rat lines. Locomotor stimulation was potentiated by the low ethanol dose in both rat lines. Ethanol treatment decreased the cerebellar and hippocampal cGMP concentrations both with and without MK-801 pretreatment in both rat lines. In situ hybridization using oligonucleotide probes specific for NMDA receptor subunit mRNAs NR1 and NR2A, B, C, and D revealed no clear differences in brain regional expression between ANT and AT rates. These results indicate that the alcohol-sensitive ANT rats are very sensitive to a low dose of ethanol in the presence of NMDA receptor antagonism, consistent with the hypothesis that this receptor system is involved in acute ethanol intoxication.

Alcoholism↗

High sensitive fluorophotometric determination of nucleic acids with Pyronine G sensitized by N,N-dimethylformamide.

In Tris-HCl buffer (pH 8.0), the fluorescence of Pyronine G emitted at 552 nm was quenched by nucleic acids when excited at 525 nm. Adding N, N-dimethylformamide (DMF) as a sensitive media can enhance the sensitivity greatly. Based on the fluorescence reactions sensitive fluorimetric methods for nucleic acids at nanogram levels were proposed. Under the optimum conditions, the calibration curves were linear in the range of 0.0032 - 2.5 microg mL(-1) for ct DNA and 0.0024 - 2.5 microg mL(-1) for hs DNA. The limits of determination were 3.2 ng mL(-1) and 2.4 ng mL(-1) respectively. This method has good selectivity and high sensitivity. It has been applied to the determination of DNA in the synthetic samples and real samples with satisfactory results.

Animals↗

Characterization of skin prick testing responses for detecting sensitization to detergent enzymes at extreme dilutions: inability of the RAST to detect lightly sensitized individuals.

We observed that a group of detergent enzyme workers with known exposure to the subtilisin enzyme, Alcalase (Novo Industries, Bagsvaerde, Denmark), exhibited percutaneous sensitivity to Savinase (Novo Industries), a microbial protease, to which there was no previous occupational exposure. This was attributed to either cross-reactivity between these enzymes or to foreign enzyme contaminants contained in the Savinase antigen. The aims of this study were to determine the range of concentrations eliciting percutaneous responses to Alcalase and to another enzyme, Rapidase (an alpha-amylase) (Gist Brocades, Belgie, Netherlands); to compare the sensitivity of RAST and skin prick testing; and to characterize the relationship between wheal size and antigen concentration. Prick testing was conducted over six log10 antigen dilutions of Alcalase and Rapidase in 30 workers with previous exposure and skin reactivity to enzymes (group 1) and compared to nonexposed control groups, which included 60 atopic subjects (group 2) and 30 nonatopic subjects (group 3). The RAST was performed with Alcalase and Rapidase antigens. The percutaneous threshold concentrations in group 1 subjects varied widely from 10(3) to 10(-3) micrograms of protein per milliliter. Of 19 group 1 workers with skin test reactivity to Alcalase, 84% had positive RAST results; 83% of 24 workers who were reactive to Rapidase had positive RAST results. It was concluded that skin prick testing is preferred over in vitro methods for longitudinal monitoring of human sensitization to workplace allergens. In addition, the data predicted that based on a known Alcalase level of 0.07% in Savinase, 26% of Alcalase-sensitized subjects could react to Savinase. An excellent correlation (r > 0.97) was found between log concentration of antigen and wheal size parameters, with the log diameter and log area performing equally as well (r > 0.98). Analysis of variance revealed that more than 60% of intragroup variation represented human variability in wheal parameters at each concentration tested, whereas at least 95% of intergroup variation was due to regression. The excellent correlations of both wheal diameter and area with antigen concentrations were attributed to the very small changes observed between test concentrations.

Allergens↗

Chemosensory function and psychological profile in patients with multiple chemical sensitivity: comparison with odor-sensitive and asymptomatic controls.

OBJECTIVE: We addressed the question if patients with multiple chemical sensitivity (MCS) differ from participants with self-reported odor sensitivity without MCS and asymptomatic controls in terms of chemosensory, cognitive, and clinical psychological endpoints. METHODS: In a clinical study 23 MCS patients, 21 participants with self-reported odor sensitivity, and 23 controls were investigated using electrophysiological and psychophysical olfactometric tests [chemosensory-event-related potentials (CSERP), olfactory thresholds, odor identification, trigeminal sensitivity]. The participants filled in a mood list, a list of complaints (BL), a Symptom Check List, a State-Trait Anxiety Inventory (STAI), and an MCS questionnaire. RESULTS: The olfactometric investigations revealed no significant differences between the groups. The MCS group reached significantly higher scores on negative mood states following odorant exposure, on health complaints, global indices, and the somatization subscale of the Symptom Check List, trait and state anxiety and symptoms, and triggering matters of the MCS questionnaire. CONCLUSIONS: Our findings reveal that neither olfactory functions, nor chemosensory or cognitive olfactory information processing are impaired in MCS patients. They rather support findings of altered psychological profile and moderate psychopathology.

Adult↗

Assessment of contrast sensitivity. Part II: The relationship between objective lens opacity and laser interferometric contrast sensitivity in the cataract patient.

The laser interferometer can effectively bypass the optics of the eye and measure retinal function in patients with immature cataracts. However, it is not known how much laser interferometric measurements are impaired by cataract density. In this study we compared objective lens opacity using the IntraOptics opacity lensmeter with contrast sensitivity (CS) measured by a Randwal He-Ne laser interferometer. Comparison of lens opacity with CS in the cataract population revealed an inverse linear relationship between objective lens opacity and retinal contrast sensitivity. Separation by cataract type showed correlation coefficients as high as -0.91 for nuclear sclerotic predominant cataracts and as low as no significant correlation for posterior subcapsular predominant cataracts. Comparisons of before and after implantation surgery contrast sensitivities (as measured by the laser interferometer) with preoperative lens opacities (as measured by the IntraOptics opacity lensmeter) quantified the extent to which laser interferometric measurements underestimated potential retinal function. We found that for all cataracts, other than posterior subcapsular predominant cataracts, potential contrast sensitivity (in decibels) was underestimated by about 0.2 to 0.3 times the opacity measured by this technique.

Adult↗

The facets of anxiety sensitivity represented in the childhood anxiety sensitivity index: confirmatory analyses of factor models from past studies.

Results of past factor analytic studies of the Childhood Anxiety Sensitivity Index and Anxiety Sensitivity Index were used to formulate hypotheses about factor models of anxiety sensitivity. Using a nonclinical sample of 767 children and adolescents and confirmatory factor analysis, hypothesized models with 2, 3, and 4 lower order factors (facets) were tested. Goodness-of-fit criteria indicated that a model with 4 facets fits these data well. Support was found for factorial invariance of the 4 facets across age and gender, using nonclinical and clinical samples. Results support a hierarchical factor model in that there was a strong general factor, explaining 71% of the variance. Findings are discussed in the context of anxiety sensitivity theory and research with children and adolescents.

Adolescent↗

Sensitization as a mechanism for multiple chemical sensitivity: relationship to evolutionary theory.

Multiple chemical sensitivity (MCS) is a disorder in humans attributed to prior chemical exposure. Sensitization is an amplification of neuronal responsiveness that elicits increased behavioral responding to stimuli, and occurs in a recently developed rat model of MCS. Rats were exposed to repeated formaldehyde (Form) and their response in three behavioral tests, including locomotor activity after a cocaine challenge, conditioned fear, and behavioral avoidance of Form, was assessed. In all three tests, rats demonstrated sensitized behaviors, implicating amplified responding within specific limbic brain regions. Evolutionary theory in the context of MCS specifies how the behavioral strategies of those with MCS are consistent with the notion that their self-perceived sense of survival and reproductive fitness may be threatened by chemical exposures. This behavior may be mediated by the same limbic brain regions that become sensitized after repeated chemical exposure in animals.

Animals↗

Dentine hypersensitivity. A study of the patency of dentinal tubules in sensitive and non-sensitive cervical dentine.

Based on the hydrodynamic theory for stimulus transmission across dentine, it would be logical to conclude that teeth exhibiting the clinical symptoms referred to as dentine hypersensitivity should have dentinal tubules open at the root surface and patent to the pulp. With the exception of studies on cut dentine, there is little direct evidence to support this conclusion. In this study, caries-free teeth with exposed cervical root areas scheduled for extraction which were classified as non-sensitive or hypersensitive after suitable stimulation were examined by scanning electron microscopy. Hypersensitive teeth showed highly significantly increased numbers of tubules per unit area (approximately 8 X) compared with non-sensitive teeth. Tubule diameters were significantly wider (approximately 2 X) in hypersensitive compared to non-sensitive teeth. The number of teeth showing the penetration of methylene blue through the zone of exposed cervical dentine was larger and the depth of penetration greater in hypersensitive teeth compared to non-sensitive teeth. The results provide further evidence that stimulus transmission across dentine in hypersensitive teeth is mediated by a hydrodynamic mechanism. An understanding of factors which open dentinal tubules would seem important if attempts to prevent or treat dentine hypersensitivity are to be successful.

Dentin↗

Modified quantitative insulin sensitivity check index is better correlated to hyperinsulinemic glucose clamp than other fasting-based index of insulin sensitivity in different insulin-resistant states.

Fasting-based index estimates of insulin sensitivity were compared with euglycemic hyperinsulinemic clamp (IS clamp) measurements in 148 subjects: normal controls (n = 46), and obese (n = 12), polycystic ovary syndrome (n = 16), first-degree relatives of type 2 diabetic (n = 17), impaired glucose tolerance (n = 28), and type 2 diabetic (n = 29) patients. The fasting-based indexes tested included log homeostasis model assessment (HOMA), the quantitative insulin sensitivity check index (QUICKI), the revised QUICKI, and a new revised QUICKI using fasting plasma glycerol. In the population studied, at 40 mU/m(2).min (n = 30) revised QUICKI (r = 0.86; P < 0.0001) and QUICKI-glycerol (r = 0.87; P < 0.0001) gave higher correlations with the IS clamp than QUICKI and log HOMA (r = 0.78 and r = -0.78; P < 0.001). For subjects tested at 75 mU/m(2).min (n = 118), comparable correlations were found for all indexes (r > 0.80; P < 0.0001). When studied in subgroups, revised QUICKI and QUICKI-glycerol give significantly higher correlations with the IS clamp than other indexes for lean control subjects studied at 40mU/m(2).min and impaired glucose tolerance subjects. We confirmed, in a large patient population with a wide range of insulin sensitivities, that no single test is superior in all groups of patients. However, QUICKI and revised QUICKI are good indexes that offer correlations similar to or higher than values obtained with log HOMA. Such indexes are simple tools to estimate insulin sensitivity appropriate for epidemiological studies.

Adolescent↗

High-sensitivity determination of birefringence in turbid media with enhanced polarization-sensitive optical coherence tomography.

Polarization-sensitive optical coherence tomography provides high-resolution cross-sectional characterization of birefringence in turbid media. Weakly birefringent biological tissues such as the retinal nerve fiber layer (RNFL) require advanced speckle noise reduction for high-sensitivity measurement of form birefringence. We present a novel method for high-sensitivity birefringence quantification by using enhanced polarization-sensitive optical coherence tomography (EPS-OCT) and introduce the polarimetric signal-to-noise ratio, a mathematical tool for analyzing speckle noise in polarimetry. Multiple incident polarization states and non-linear fitting of normalized Stokes vectors allow determination of retardation with +/-1 degrees uncertainty with invariance to unknown unitary polarization transformations. Results from a weakly birefringent turbid film and in vivo primate RNFL are presented. In addition, we discuss the potential of EPS-OCT for noninvasive quantification of intracellular filamentous nanostructures, such as neurotubules in the RNFL that are lost during the progression of glaucoma.

Algorithms↗

A colorimetric in vitro drug sensitivity assay for Plasmodium falciparum based on a highly sensitive double-site lactate dehydrogenase antigen-capture enzyme-linked immunosorbent assay.

We report a double-site enzyme-linked lactate dehydrogenase immunodetection assay (DELI), a highly sensitive antigen-capture enzyme-linked immunosorbent assay, which proved to be more sensitive for the detection of Plasmodium falciparum than thick blood smears, as sensitive as the polymerase chain reaction, and probably more reliable. This technique can help to detect infra-microscopic parasitemias (one parasite in 10(6)-10(8) red blood cells) from biological samples, and being quantitative, provide a fast substitute to thick smears for epidemiologic purposes. The technique can also be used to measure the in vitro drug sensitivity of P. falciparum with greater ease, much greater speed, and simpler equipment than that required for the isotopic microtest. Results obtained with four antimalarial drugs upon 16 strains closely paralleled those obtained by the isotopic assay (R = 0.95). In contrast with the latter, much lower parasite densities could be tested in the DELI assay (as low as 0.005%), thereby extending the number of isolates that can be investigated. The ease of implementation and low cost of the DELI-microtest may contribute to a revived interest in using in vitro methods to survey resistance to antimalarial drugs, so as to better predict future in vivo drug failures and provide public health recommendations.

Animals↗

[Sensitivity loss in short wavelength sensitive cones in myopic eyes].

To investigate early changes of visual function in high myopic eyes, the relative sensitivity of the short-wavelength sensitive cones (s-cones) was measured by flash-evoked cortical potentials and electroretinograms and their relation with refractive error and axial length of the eyes was studied. A Ganzfeld stimulus was employed. The subjects were all under 37 years of age and were divided into 3 groups according to the magnitude of the refractive error as follows; 1) emmetropia/low myopia group (10 eyes), between +0.50 D and -3.00 D; 2) medium myopia group (8 eyes), between -3.25 D and -6.00 D; and 3) high myopia group (10 eyes), over -6.25 D. Their corrected visual acuities were better than 1.0 and their color vision was normal according to Panel D-15 and SPPII. They had no other ophthalmological abnormalities except tigroid fundus. The s-cone sensitivity was significantly correlated with the refractive error and axial length (p < 0.001) and was reduced prior to ophthalmoscopic retinal changes in the medium and high myopia groups. This indicates that the site of s-cone sensitivity loss in myopia is in the retinal.

Adolescent↗

No deterioration in insulin sensitivity, but impairment of both pancreatic beta-cell function and glucose sensitivity, in Japanese women with former gestational diabetes mellitus.

To identify the primary pathogenic factors involved in the development of Type 2 diabetes mellitus (DM), we studied Japanese women with former gestational diabetes mellitus (GDM) who are at risk for the later development of Type 2 DM. We used the minimal model analysis derived from frequently sampled intravenous glucose tolerance test (FSIGT). The subjects consisted of eight non-obese women with a history of GDM and eight non-obese normal women as control subjects. The 75 g oral glucose tolerance test (75 g OGTT) performed within 6 months of delivery confirmed that all the subjects with former GDM had a normal glucose tolerance. Insulin sensitivity (SI) derived from the minimal model analysis was not different between the two groups. Glucose effectiveness at zero insulin (GEZI), reflecting tissue glucose sensitivity, was significantly lower in former GDM patients than in control subjects (1.18+/-0.34 vs 2.26+/-0.29 x 10(-2) min(-1), p < 0.05). The early phase insulin secretion found in FSIGT was markedly reduced to 56% of that observed in control subjects (1250+/-87.4 vs 2223+/-304.3 pmol l(-1) min, p < 0.01). Our results indicate that in former GDM patients, who are japanese and non-obese, impairment of the acute insulin response to glucose and a decrease in tissue glucose sensitivity rather than insulin sensitivity are the primary pathogenic factors involved.

Blood Glucose↗

Sensitizing properties of new N-(2-arylalkyl) propionamides in drug-sensitive and multidrug-resistant tumor cells.

The propionamides 5 and 6 have been synthesized and tested for stimulation of antitumor drug activity. 5 and 6b increase vincristine cytotoxicity in drug-sensitive murine tumor cells; 5 also increases the toxicity in multidrug resistant cells. Dissimilar trends in sensitive and resistant cells have been observed for the stimulating activity of several propionamides of this family and structurally related verapamil with their molar refractivity, suggesting different size requirements for the sensitizers in sensitive and resistant cells.

Amides↗

Wakefulness-inducing area in the brainstem excites warm-sensitive and inhibits cold-sensitive neurons in the medial preoptic area in anesthetized rats.

Sleep-wakefulness and body temperature are known to influence each other. The body temperature rises during wakefulness and falls during sleep. The midbrain reticular formation is one of the areas in the brainstem that induces wakefulness, while the preoptico-anterior hypothalamic area is the main thermoregulatory center in the brain. In order to understand the neural mechanism for simultaneous regulation of these functions we hypothesized that the wakefulness area in the brainstem is likely to have an opposite influence on warm- and cold-sensitive neurons in the preoptico-anterior hypothalamic area. Hence, first, the wakefulness-inducing area was identified in the brainstem by stimulating the site with high-frequency rectangular wave electrical pulses (100 Hz, 100 microA, 200 microsec for 5-8 sec) in freely behaving chronically prepared experimental rats. Then, single neuronal activity from the medial preoptico-anterior hypothalamic area was recorded and their thermosensitivity was established. Thereafter, the influence of such a confirmed wakefulness-inducing area in the brainstem on the responsiveness of the single neuronal activity of predetermined warm- and cold-sensitive neurons as well as on temperature-insensitive neurons was studied by overlapping stimulus (1 Hz, 500 microA, 200 microsec) bound responses. It was observed that the warm-sensitive neurons were excited and the cold-sensitive neurons were inhibited by stimulation of the wakefulness-inducing area in the brainstem. Most of the temperature-insensitive neurons remained unaffected. The results confirm our hypothesis and help in understanding the mechanism of simultaneous modulation of body temperature in association with changes in wakefulness at the single neuronal level.

Anesthetics, Intravenous↗