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PubMed1977

Reaginic antibody production to Ascaris suum allergen, ASC-1. I. The function of glutaraldehyde-polymerized antigen in the induction of reaginic (IgE) antibodies in the rat.

Rats immunized with a purified Ascaris suum allergen (Asc-1) or with its dinitrophenylated derivate (DNP-Asc-1) produced high levels of reaginic (IgE) antibodies. A second injection of antigen given 30 days later did not result in an anamnestic IgE antibody response. Immunization of adult-thymectomized, lethally-irradiated and bone-marrow reconstituted (ATxB) rats with soluble Asc-1 or DNP-Asc-1 failed to stimulate reaginic antibody production. The administration of glutaraldehyde-polymerized antigen induced in some but not all ATxB rats, low but detectable levels of IgE antibodies. These levels increased following a second injection of nonpolymerized antigen in A1 (OH)3 gels. Priming of animals with polymerized carrier and Bordetella pertussis did not stimulate a primary anticarrier IgE response but led to an enhanced antihapten IgE response following the administration of soluble DNP-Asc-1 in A1(OH)3. The results are consistent with the notion that a sharply reduced but clearly functional T-derived helper cell population could be triggered by the polymerized but not by the soluble form of the immunogen.

Allergens↗

PubMed2006

ASC-US and high-risk HPV testing: performance in daily clinical practice.

Data are beginning to accrue on high-risk HPV DNA testing in patients with ASC-US on cervical cytology. We report on our experience at the University of Wisconsin Hospital and Clinics. From February 2002 through December 31, 2005 (3 yr, 11 mo), the cytopathology laboratory processed 49,599 Pap Tests, of which 1,792 (3.6%) were diagnosed as ASC-US. Six hundred and seventy two (37.5%) of these cases were processed for high-risk HPV genotypes using the Digene Hybrid Capture II method. Of these cases, 266 (39.6%) were positive for high-risk HPV genotypes, 11 (1.6%) were equivocal, and 395 (58.8%) were negative. Biopsy follow-up was available for 127 (47.7%) of the 266 cases, of which 66 (52%) were negative, 46 (36.2%) showed CIN I, 9 (7.1%) were CIN II, and 6 (4.7%) were CIN III. Of the remaining 139 (52.3%) cases, 86 (62%) had follow-up Pap Tests, of which 57 (66.3%) were negative, 15 (17.4%) were ASC-US, 12 (15%) were low-grade squamous intraepithelial lesions, and 2 (2.3%) were high-grade squamous intraepithelial lesions; 53 (38.1%) were lost to follow-up. In combination, 90 (42.25%) of the 213 cases with follow-up showed atypia or above after a diagnosis of ASC-US; of which 58 (64%) were low-grade lesions and 17 (19%) were high-grade lesions. Our laboratory's reported high-risk HPV positivity is comparable to recent reports in the literature on its use in daily clinical practice. In addition, cervical abnormalities were found in a significant proportion of the cases.

Adolescent↗

PubMed1993

The Asc locus for resistance to Alternaria stem canker in tomato does not encode the enzyme aspartate carbamoyltransferase.

The fungal disease resistance locus Alternaria stem canker (Asc) in tomato has been suggested to encode the enzyme aspartate carbamoyltransferase (ACTase). To test this hypothesis a segment of the tomato ACTase gene was amplified by the polymerase chain reaction (PCR) using degenerate primers. The PCR product obtained was subsequently used to isolate an ACTase cDNA clone. Restriction fragment length polymorphism (RFLP) linkage analysis showed that the ACTase gene and the Asc locus do not cosegregate. RFLP mapping positioned the ACTase gene on chromosome 11, while the Asc locus is located on chromosome 3. These results exclude the possibility that the ACTase protein is encoded by the Asc locus.

Alternaria↗

PubMed2005

Role of charged and hydrophobic residues in the oligomerization of the PYRIN domain of ASC.

Apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is an adaptor protein composed of two homophilic protein-protein interaction domains, a PYRIN domain (PYD) and a caspase recruitment domain. PYD-dependent oligomerization of ASC is thought to play a crucial role in formation of a molecular platform, the inflammasome, which activates caspase-1. When expressed in cells, the PYD of ASC was shown to form cytoplasmic filaments through self-association. Over 70 single point mutants were analyzed for filament formation in cells expressing the mutant proteins. The set of mutations comprised every single amino acid residue with a charged side chain (Arg, Lys, Asp, and Glu) and a large hydrophobic side chain (Ile, Leu, Met, Phe, Pro, and Val). Filament formation of the ASC PYD was prevented by mutation of Lys21, Leu25, Lys26, Pro40, Arg41, Asp48, and Asp51 of helices 2, 3, and 4. These data identify a coherent interaction surface, establishing a molecular model of PYD-PYD complexes with an important role for charge-charge interactions.

Amino Acid Sequence↗

PubMedDate not supplied

Influence of HMB (beta-hydroxy-beta-methylbutyrate) on antibody secreting cells (ASC) after in vitro and in vivo immunization with the anti-Yersinia ruckeri vaccine of rainbow trout (Oncorhynchus mykiss).

In practice, protection of fish against disease by immunization is of limited effectiveness. Therefore, research is concentrated on how to improve the potency and efficacy of vaccines and how to optimally activate the cell-mediated immunity and the specific antibody response. In the present study, the influence of HMB (beta-Hydroxy-beta-methylbutyrate) on the antibody secreting cells (ASC) after both in vitro and in vivo immunization of rainbow trout (Oncorhynchus mykiss) with the anti-yersiniosis vaccine was studied. For in vitro immunization, the spleens from 160 fish were sampled and placed each in 35 mm sterile wells with medium containing HMB at concentrations of 0, 0.1, 1, 5, 10, 25, 50 or 100 microg/mL of medium. The spleens from 80 fish were injected with the vaccine and incubated at 14 degrees C for 10 days. For the in vivo study, fish were fed pellets containing HMB at doses of 0, 10, 25 and 50 mg/kg bw per day. After 2 weeks of HMB supplementation, the fish were immunized by intraperitoneal injection of the vaccine. At 7, 14, 18, 21, 28 and 35 days after immunization, pronephros were taken from 10 fish in each group for testing. When analyzed by the ELISPOT assay, HMB increased the number of splenic ASC after in vitro immunization at concentrations between 10 and 100 microg/mL (P < 0.05). Dietary HMB also increased the number of total and specific ASC when the fish were vaccinated in vivo. In conclusion, the results of the present study showed that HMB increases the levels of specific ASC after both in vitro and in vivo immunization of rainbow trout with the anti-Yersinia ruckeri vaccine.

Animals↗

PubMed1998

The standardized psychometric assessment of altered states of consciousness (ASCs) in humans.

The APZ questionnaire was developed in order to explore hypotheses on ASCs. First -- in a series of 11 experiments using different induction methods on N = 393 healthy subjects -- the hypothesis was tested that ASCs have major dimensions in common irrespective of the mode of their induction. In the International Study on Altered States of Consciousness (ISASC) the external validity of the experimental results was assessed. The ISASC was carried out on a total of N = 1133 subjects in six countries. The main results of the experimental studies were corroborated in the field studies. The results can be summarized as follows: the common denominator of ASCs is described by three oblique dimensions, designated as "Oceanic Boundlessness (OSE)", "Dread of Ego Dissolution (AIA)" and "Visionary Restructuralization (VUS)". The reliability and validity of the scales are satisfactory. Tested versions of the APZ scales are available in English (UK, USA), German, Italian and Portuguese. Psychometrically as yet untested versions exist in Dutch, Finnish, French, Greek, Spanish and Russian. The APZ questionnaire has become the international standard for the assessment of ASCs, thus helping to integrate research. A psychometrically improved version exists in German (OAV questionnaire). The BETA questionnaire, which measures the dimensions "Vigilance Reduction (VIR)" and "Auditive Alteration (AVE)" is also available in German. These dimensions are most likely etiology-dependent.

Clinical Trials as Topic↗

PubMed2002

PYPAF1, a PYRIN-containing Apaf1-like protein that assembles with ASC and regulates activation of NF-kappa B.

The PYRIN domain is a recently identified protein-protein interaction domain that is found at the N terminus of several proteins thought to function in apoptotic and inflammatory signaling pathways. We report here that PYPAF1 (PYRIN-containing Apaf1-like protein 1) is a novel PYRIN-containing signaling protein that belongs to the nucleotide-binding site/leucine-rich repeat (NBS/LRR) family of signaling proteins. The expression of PYPAF1 is highly restricted to immune cells, and its gene maps to chromosome 1q44, a locus that is associated with the rare inflammatory diseases Muckle-Wells syndrome and familial cold urticaria. To identify downstream signaling partners of PYPAF1, we performed a mammalian two-hybrid screen and identified ASC as a PYRIN-containing protein that interacts selectively with the PYRIN domain of PYPAF1. When expressed in cells, ASC recruits PYPAF1 to distinct cytoplasmic loci and induces the activation of NF-kappaB. Furthermore, coexpression of PYPAF1 with ASC results in a potent synergistic activation of NF-kappaB. These findings suggest that PYPAF1 and ASC function as upstream activators of NF-kappaB signaling.

Amino Acid Sequence↗

PubMed2003

Active Sequences Collection (ASC) database: a new tool to assign functions to protein sequences.

Active Sequences Collection (ASC) is a collection of amino acid sequences, with an unique feature: only short sequences are collected, with a demonstrated biological activity. The current version of ASC consists of three sections: DORRS, a collection of active RGD-containing peptides; TRANSIT, a collection of protein regions active as substrates of transglutaminase enzyme (TGase), and BAC, a collection of short peptides with demonstrated biological activity. Literature references for each entry are reported, as well as cross references to other databases, when available. The current version of ASC includes more than 800 different entries. The main scope of this collection is to offer a new tool to investigate the structural features of protein active sites, additionally to similarity searches against large protein databases or searching for known functional patterns. ASC database is available at the web address http://crisceb.unina2.it/ASC/ which also offers a dedicated query interface to compare user-defined protein sequences with the database, as well as an updating interface to allow contribution of new referenced active sequences.

Amino Acid Sequence↗

PubMed2006

Atypical squamous cells of undetermined significance, human papillomavirus, and cervical intraepithelial neoplasia 2 or 3 in adolescents: ASC-US, age, and high-grade cervical neoplasia.

OBJECTIVE: To determine rates of and factors associated with cervical intraepithelial neoplasia (CIN) 2 or 3 among women with atypical squamous cells of undetermined significance (ASC-US) and oncogenic human papillomavirus (HPV) infection. MATERIALS AND METHODS: This is a retrospective review of 357 women with ASC-US and known high-risk HPV infection. To be eligible for inclusion, patients had to have subsequently undergone colposcopic-directed cervical biopsy and/or endocervical sampling. Univariate and multivariable regression analyses were used to estimate crude and adjusted odds ratios of CIN 2 or 3 by risk factors associated with HPV and CIN. RESULTS: Thirty-three of 215 (15%) women with ASC-US and oncogenic HPV who underwent histological sampling demonstrated CIN 2 or 3. In the adjusted model, only nulliparity (adjusted odds ratio = 2.2, 95% CI = 1.0-4.8) retained a marginal association with the outcome. No significant age-related differences in the prevalence of high-grade disease emerged. The high-grade lesions seen in adolescents consisted of single, small lesions. CONCLUSIONS: In the light of these findings, and in conjunction with the high rate of oncogenic HPV infection among adolescents with ASC-US, both reflex HPV testing and immediate colposcopy do not appear to be of benefit in the initial management of teenagers with this cytological diagnosis.

Adolescent↗

PubMed1975

Amyloid-related serum component (protein ASC) IN LEPROSY PATIENTS.

The presence of amyloid-related serum component, protein ASC, in serum samples from 63 leprosy patients was investigated. Protein ASC was detected in 38% of the patients. A correlation to the disease spectrum of leprosy was apparent: polar lepromatous cases, 64% positive; borderline lepromatous, 50%; borderline tuberculoid, 36%; subpolar tuberculoid, 17%; and polar tuberculoid, negative. Antibody activity against the a antigen of Mycobacterium leprae was also determined, showing a similar correlation to the disease spectrum. Serum samples from 23 apparently healthy Ethiopians serving as controls showed a protein ASC incidence of 22%. This figure is significantly higher than the frequency found by others among healthy Norwegian blood donors. Immunoglobulin M levels among patients were elevated in the borderline lepromatous and poplar lepromatous groups. The three tuberculoid groups did not differ in this respect from the control group but were all elevated as compared to a normal Caucasian serum pool. Although raised immunoglobulin M levels seemed to parallel increased frequencies of protein ASC in the patient groups as well as in controls, this correlation might be only secondary to a primary derangement in T-cell function.

Adult↗

PubMed2002

Multiple developmental defects derived from impaired recruitment of ASC-2 to nuclear receptors in mice: implication for posterior lenticonus with cataract.

ASC-2, a recently isolated transcriptional coactivator molecule, stimulates transactivation by multiple transcription factors, including nuclear receptors. We generated a potent dominant negative fragment of ASC-2, encompassing the N-terminal LXXLL motif that binds a broad range of nuclear receptors. This fragment, termed DN1, specifically inhibited endogenous ASC-2 from binding these receptors in vivo, whereas DN1/m, in which the LXXLL motif was mutated to LXXAA to abolish the receptor interactions, was inert. Interestingly, DN1 transgenic mice but not DN1/m transgenic mice exhibited severe microphthalmia and posterior lenticonus with cataract as well as a variety of pathophysiological phenotypes in many other organs. Our results provide a novel insight into the molecular and histopathological mechanism of posterior lenticonus with cataract and attest to the importance of ASC-2 as a pivotal transcriptional coactivator of nuclear receptors in vivo.

Amino Acid Motifs↗

PubMed1986

System ASC and sodium-independent neutral amino acid transport in muscle of uremic rats.

Neutral amino acids are transported by systems A, ASC, and L. In the previous companion study we demonstrated that 2-(methylamino) isobutyrate (MeAIB) is a specific substrate for system A in muscle and that stimulation of system A by physiological concentrations of insulin is preserved in acute uremia (ARF). Insulin-stimulated uptake of the nonspecific probes cycloleucine and alpha-aminoisobutyrate (AIB) is reportedly blunted by uremia; the cause of this and whether transport by systems ASC and L is defective are unknown. In this study we examined these questions using incubated epitrochlearis muscles from normal fed, ARF, and sham-operated control (SO) rats. System ASC was studied by measuring AIB and cycloleucine uptake in the presence of inhibitors of systems A and L, MeAIB and 2-amino-2-norbornane carboxylic acid (BCH), respectively. System L was defined as sodium-independent uptake suppressible by BCH. Excess MeAIB completely inhibited insulin-stimulated AIB and cycloleucine uptake, indicating that system A is the only insulin-responsive neutral amino acid carrier in muscle. In ARF and SO mucles both AIB and cycloleucine uptake were indistinguishable in the absence or presence of insulin. Moreover, ARF caused no detectable abnormality in transport by systems ASC and L.

Acute Kidney Injury↗

PubMed1991

Characterization of threonine transport into a kidney epithelial cell line (BSC-1). Evidence for the presence of Na(+)-independent system asc [corrected].

The transport routes for threonine in a primate kidney epithelial cell line (BSC-1) grown as monolayer in continuous cell culture were studied. We discovered at least four different transport systems for threonine uptake. The Na(+)-dependent route shows biphasic kinetics with a low and high affinity parameter. The apparent kinetic constants for Km1 and Km2 were 0.3 and 36 mM with apparent Vmax values of 6.3 and 90 nmol/mg protein/min, respectively. The high affinity, low Km component resembles system ASC activity, with respect to substrate selectivity. The Na(+)-independent route also exhibits biphasic kinetics. A high affinity component (apparent Km of 1.0 mM, and apparent Vmax of 7.2 nmol/mg protein/min) is sensitive to inhibition by leucine and the aminoendolevo-rotatory isomer of 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid, suggesting participation by system L. The low affinity component (apparent Km of 10.2 mM, and apparent Vmax of 71 nmol/mg protein/min) was specifically inhibited by threonine, serine, and alanine and could be assigned to system asc. The discrimination between system L and asc is based upon differences in pH sensitivity, trans stimulation, and Ki values. In addition, the effects of harmaline, a suspected sodium transport site inhibitor, have been studied. Harmaline noncompetitively inhibited Na(+)-dependent threonine uptake but had no effect on Na(+)-independent transport of threonine. This report is the first to present evidence for the presence of system asc in renal epithelial cells. The physiological and biochemical significance of our findings are discussed.

Animals↗

PubMed2025

Evolutionary and Functional Analysis of Caspase-8 and ASC Interactions to Drive Lytic Cell Death, PANoptosis.

Caspases are evolutionarily conserved proteins essential for driving cell death in development and host defense. Caspase-8, a key member of the caspase family, is implicated in nonlytic apoptosis, as well as lytic forms of cell death. Recently, caspase-8 has been identified as an integral component of PANoptosomes, multiprotein complexes formed in response to innate immune sensor activation. Several innate immune sensors can nucleate caspase-8-containing PANoptosome complexes to drive inflammatory lytic cell death, PANoptosis. However, how the evolutionarily conserved and diverse functions of caspase-8 drive PANoptosis remains unclear. To address this, we performed evolutionary, sequence, structural, and functional analyses to decode caspase-8's complex-forming abilities and its interaction with the PANoptosome adaptor ASC. Our study distinguished distinct subgroups within the death domain superfamily based on their evolutionary and functional relationships, identified homotypic traits among subfamily members, and captured key events in caspase evolution. We also identified critical residues defining the heterotypic interaction between caspase-8's death effector domain and ASC's pyrin domain, validated through cross-species analyses, dynamic simulations, and in vitro experiments. Overall, our study elucidated recent evolutionary adaptations of caspase-8 that allowed it to interact with ASC, improving our understanding of critical molecular associations in PANoptosome complex formation and the underlying PANoptotic responses in host defense and inflammation. These findings have implications for understanding mammalian immune responses and developing new therapeutic strategies for inflammatory diseases.

Caspase 8↗

PubMed2003

Regulation of L-alanine transport systems A and ASC by cyclic AMP and calcium in a reptilian duodenal model.

The regulation of neutral amino acid transport by cyclic AMP (cAMP) and calcium across the isolated duodenum of the lizard Gallotia galloti has been studied under short-circuit conditions. Active L-alanine transport was stimulated by forskolin, theophylline and dibutyryl cyclic AMP (db-cAMP). All these agents increased transmural potential difference (PD) and short-circuit current (I(sc)) in a manner consistent with the activation of a chloride secretory pathway. Both forskolin and theophylline increased intracellular cAMP levels in the lizard duodenal mucosa. Addition of calcium ionophore A23187 rapidly reduced mucosa-to-serosa L-alanine fluxes and diminished net L-alanine transport. Despite the reduction of alanine fluxes by A23187, transepithelial PD and I(sc) values were increased by the ionophore. Analyses of the responses of isolated transport pathways indicated that the Na(+)-independent L-alanine transport system was unaffected by db-cAMP or calcium ionophore. By contrast, Na(+)-dependent transport activities were profoundly modified by these agents. Thus, while system A [alpha-methylamino-isobutiric acid (MeAIB)-transporting pathway] was stimulated by increased calcium, system ASC activity was nearly abolished. Calcium ionophore also potentiated the electrogenic response of system A. Forskolin strongly stimulated system ASC activity but left system A activity unchanged. Activation of system ASC by forskolin was clearly electroneutral, as pre-incubation of the tissues with the chloride channel blocker diphenylamine-2-carboxilic acid (DPC) completely prevented forskolin-induced transepithelial electrical responses. It is concluded that intracellular messengers cAMP and calcium oppositely modulate active Na(+)-dependent (L)-alanine transport in the lizard intestine. The different sensitivity exhibited by individual transport pathways may well account for the changes observed in overall alanine transport.

Alanine↗

PubMed2006

The Acute Stress Checklist for Children (ASC-Kids): development of a child self-report measure.

This project aimed to develop and validate a brief, practical self-report measure of acute stress disorder (ASD) reactions (including ASD diagnostic criteria) in children and adolescents. A draft measure was reviewed for content validity by an expert panel, piloted with youth, and revised for clarity and coverage of key constructs. The new 29-item Acute Stress Checklist for Children (ASC-Kids) was administered to 176 youth age 8 to 17 who had a recent injury or intensive care unit admission. The ASC-Kids demonstrated strong test-retest reliability and internal consistency, as well as concurrent and predictive validity with other traumatic stress measures. Results of exploratory factor analyses were consistent with current conceptualizations of acute traumatic stress reactions. The ASC-Kids is a promising measure of child acute stress reactions.

Adolescent↗

PubMed1999

A Na(+)-dependent system A and ASC-independent amino acid transport system stimulated by glucagon in rat hepatocytes.

The effects of glucagon on amino acid transport in rat hepatocytes are not fully understood. We examined the effect of this hormone on alanine, serine and cysteine preferring system (system ASC)-mediated amino acid transport in rat hepatocyte monolayers using 2-aminoisobutyric acid (AIB) and l -cysteine. Glucagon induced a time and protein synthesis-dependent stimulation of Na(+)-dependent alanine preferring system (system A)-independent AIB transport. The glucagon-induced increase in transport activity was not modified by substrate starvation and not related to changes in the intracellular pool of amino acids. Glucagon did not modify system ASC activity measured by l -cysteine. Therefore the transport activity of AIB independent of system A stimulated by glucagon cannot be attributed to system ASC. This suggests a Na(+)-dependent transport system in rat hepatocytes not identified until now.

Amino Acids↗

PubMed2001

Murine ortholog of ASC, a CARD-containing protein, self-associates and exhibits restricted distribution in developing mouse embryos.

ASC (apoptosis-associated speck-like protein containing a CARD) was first identified as a cytosolic soluble protein that forms insoluble aggregates and enhances etoposide-induced apoptosis. We have cloned a murine ortholog of ASC (mASC) comprising 193 amino acids with a well-conserved pyrin N-terminal homology domain and caspase recruitment domain (CARD). mASC fused with green fluorescent protein appeared as a speck in transfected COS-7 cells and showed self-association. We analyzed mASC gene expression in developing embryos by in situ hybridization and found it to have a restricted distribution in mouse embryos. At E9.5, mASC was strongly expressed in the telencephalon, thalamic areas of the diencephalon, heart, and liver. Northern blotting analysis revealed that the mASC gene was expressed ubiquitously in multiple organs in adult mice. These findings indicate that mASC shows conservation of not only the primary structure of human ASC but also the ability to aggregate and has some similarity in its distribution to other CARD-containing molecules, including the apoptosis regulator Apaf-1.

Amino Acid Sequence↗