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Enzyme
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Query: UMLS:C0042961 (
volvulus
)
4,305
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
A
PEG
-ELISA was used to demonstrate parasite specific immune complexes in a significant proportion (25/26) of Onchocerca volvulus infection sera from Sierra Leone. The parasite antigen was detected using a peroxidase-conjugated rabbit serum raised to the bovine parasite O. gibsoni. Controls including European control serum, endemic control serum and Rh+ sera gave consistently low readings. Characterization of the parasite component in the immune complexes by Western blotting demonstrated a heat stable antigen of M(r) 46,000. This antigen was not present in the circulating immune complexes (CIC) prepared from patients with Wuchereria bancrofti infection, but a cross-reactive molecule of the same size was weakly recognized in the CIC of Loa loa and Mansonella perstans infected patients. No association between the level of parasite specific CIC and clinical disease was observed in the O.
volvulus
patients.
...
PMID:Identification and characterization of a parasite antigen in the circulating immune complexes of Onchocerca volvulus infected patients. 128 95
In the present study, the enzyme acetylcholinesterase (AChE) from filarial parasites was identified in sera from humans infected with Onchocerca
volvulus
as well as in Mastomys natalensis infected with Brugia pahangi. The enzyme was present in immune complexes precipitated with cold 4%
polyethylene glycol
. The infected sera showed 3-4 times more AChE activity than did normal sera, and enzyme activity could be demonstrated in 5% polyacrylamide gels by specific staining. The enzyme from infected serum showed 3 times more activity when acetylthiocholine was used as the substrate as compared with butyrylthiocholine, whereas the enzyme activity present in normal serum was low and did not show this substrate specificity. Immunoprecipitation assays confirmed the presence of anti-AChE antibodies in the infected serum. The enzyme was further analysed by enzyme-linked immunosorbent assay and immunoblotting with rabbit antibodies to B. malayi AChE. Immunoblotting of the B. pahangi-infected serum revealed two closely located bands at about 200 kDa and one 95-kDa band, whereas in O.
volvulus
-infected serum, only one specific band was observed at about 200 kDa. The identification of parasite AChE may be particularly useful for diagnosis of the disease or for the study of the involvement of this enzyme in the host-parasite relationship.
...
PMID:Identification of circulating parasite acetylcholinesterase in human and rodent filariasis. 148 Jun 4
This study identified and characterized parasite antigens in sera from humans infected with Onchocerca
volvulus
. Immune complexes were precipitated from human sera with
polyethylene glycol
and analyzed by immunoblot with rabbit antibodies to O.
volvulus
. A 23-kDa parasite antigen was detected in sera from 17 of 23 Nigerian onchocerciasis patients and 5 of 10 endemic controls. Other parasite antigens with apparent molecular masses of 62, 66, and 70 kDa were less consistently observed. These antigens were not present in Nigerian or US nonendemic control sera, in sera from patients with various other parasitic infections, or in sera from US patients with autoimmune diseases. Biochemical studies indicated that these antigens are nonglycosylated acidic proteins that do not contain phosphorylcholine. These antigens may be useful as targets for improved diagnostic tests for onchocerciasis based on parasite antigen detection.
...
PMID:Circulating immune complex-associated parasite antigens in human onchocerciasis. 223 Feb 40
The viability and drug responses of cryopreserved adult Onchocerca have been examined in vitro. Male worms were cryopreserved in liquid nitrogen (-196 degrees C) using
ethanediol
as a cryoprotectant in a 2-step incubation procedure. After thawing, 85-90% of O. gutturosa males were normally motile. These motile worms were evaluated for viability using 4 measurements (long-term motility/survival in culture; [U-14C]adenine uptake and leakage; glucose utilization; MTT-formazan colorimetry) and were no different from unfrozen controls. Subsequent experiments demonstrated that the motility responses of cryopreserved worms exposed to the antifilarial drugs ivermectin, CGP 6140 and levamisole were virtually identical to unfrozen controls. Some success was also obtained with this technique in cryopreserving O.
volvulus
males, with 2 thawed specimens surviving in culture for 93 and 106 d respectively. Following collagenase isolation, female worms were cryopreserved in medium +10% serum without protectant at -79 degrees C. A batch of 8 female O. gutturosa were all motile when thawed 14 d later, with a mean survival time (based on 5 specimens) of 71 d (range 60-90). However, a batch of worms transferred from -79 degrees C to -196 degrees C were badly damaged when thawed. Female O.
volvulus
were cryopreserved at -79 degrees C in Guatemala and sent by air freight on solid CO2 to the UK. Most specimens were active when thawed. Survival of motile specimens ranged from 7 to 272 d in culture. It is concluded that these techniques are of practical value for the storage and transportation of adult Onchocerca.
...
PMID:Onchocerca gutturosa and O. volvulus: studies on the viability and drug responses of cryopreserved adult worms in vitro. 261 29
A range of culture conditions were examined to optimize parasite maintenance. Using male worms in a cell-free system, good results were obtained with medium NCTC 135 + 10% inactivated calf serum (IFCS) in an atmosphere of 95% N2/5% CO2 (median survival time 45 days). Survival was increased to 6-7 months using medium MEM + 10% IFCS + LLCMK2 (monkey kidney) feeder cells in a gas phase of 5% CO2 in air. Worms exposed to collagenase solution (5 mg/ml) were subsequently less motile and survived shorter periods compared to unexposed controls. The drug responses of worms (in vitro) were examined using 13 antiparasitic compounds. Ivermectin and CGP 6140 were among the most active, with the majority of drugs significantly affecting motility levels at a concentration of 5 x 10(-5) M or less. This system may provide useful information on the intrinsic activity of new compounds. A technique was developed for the successful cryopreservation of males in liquid nitrogen using
ethanediol
as a cryoprotectant in a 2-step incubation procedure, thereby enabling the long-term storage and transportation of worms. In conclusion, the common bovine parasite O. gutturosa provides a practical alternative for research in the absence of O.
volvulus
.
...
PMID:The development of a laboratory model for onchocerciasis using Onchocerca gutturosa: in vitro culture, collagenase effects, drug studies and cryopreservation. 285 98
This report describes the presence of circulating Onchocerca
volvulus
antigens (COA) in sera of patients with onchocerciasis. By using a double diffusion immunoelectrophoresis method, COA could be detected in 24 of 77 sera analyzed (31%). In contrast, when more sensitive assays such as the radioimmunoprecipitation-
PEG
assay or sandwich radioimmunoassay were used to detect COA, about 75% of the sera from O.
volvulus
-infected patients were found positive; moreover, a highly significant correlation between the two assays was observed. The parasite specificity of the COA was demonstrated directly by identity reaction with a component of O.
volvulus
somatic antigens. COA was never found when hyperimmune antisera against other parasite antigenic extracts were used instead of anti-O.
volvulus
hyperimmune serum. However, when anti-O,
volvulus
hyperimmune serum was used against sera obtained from patients infected with various other helminths we found a cross-reactivity between COA and the circulating antigens of other human filarids (Wuchereria bancrofti, Loa loa, Brugia malayi), but not with other nematode or trematode parasites (Ascaris lumbricoides, Schistosoma mansoni, Fasciola hepatica). Further immunoelectrophoretic studies demonstrated one precipitin are localized in the cathodic region which seemed specific for COA, which raises the possibility of preparing a monospecific hyperimmune serum to circumvent cross-reactivities.
...
PMID:Detection of circulating antigens in onchocerciasis. 679 90
A monoclonal antibody of the IgM class recognizing Onchocerca
volvulus
circulating antigen (COA) was obtained. This monoclonal antibody was used in a radioimmunoprecipitation-
PEG
assay (RIPEGA) to detect circulating antigen in onchocerciasis patients' sera. COA could be detected in 63 (80%) of the 79 African patient sera tested, and in 126 (76%) of the 164 Indian (Venezuela) sera studied. There was no direct correlation between the presence of COA detected in the patient serum and the level of microfilarodermia. The RIPEGA using this monoclonal antibody detected COA in 91% of children under 10 years old, whereas the microfilarodermia in this group was positive in only 52% of the cases. The specificity of this test is improved compared to the results obtained with polyclonal antibodies. Immunofluorescence studies suggest that the COA might be located in the microfilaria cuticle.
...
PMID:Onchocerca volvulus: detection of circulating antigen by monoclonal antibodies in human onchocerciasis. 685 99
Improved methods for diagnosis of onchocerciasis are needed. We have recently identified immune complex-associated parasite antigens in sera from onchocerciasis patients. The goal of this study was to produce monoclonal antibodies to these antigens that might be used in antigen detection assays. Two monoclonal antibodies (OV-1 and OV-5) that bind to parasite antigens in immunoblots of
PEG
-precipitated immune complexes from human onchocerciasis sera and to corresponding antigens in adult worm extracts and excretory-secretory products were produced. The target epitopes of the monoclonals are heat stable, resistant to trypsin, and destroyed by Pronase. The two monoclonals produce similar but not identical patterns of binding to immunoblots of Onchocerca
volvulus
adult worm antigen with major bands at 43-47, 58-63, and 70 kDa. OV-1 and OV-5 appear to bind to two distinct but closely related epitopes, neither of which is phosphorylcholine. Immunoelectron microscopy showed that the epitopes recognized by these monoclonals are widely distributed in adult female worms, but concentrated in the uterus and intestine. Antigen assays based on these antibodies detected parasite antigen in 9 of 14 sera from onchocerciasis patients, but significant background signal was detected in some nonendemic human sera. Thus, although this study has provided new information on parasite antigens in sera from onchocerciasis patients, additional work will be needed to achieve the goal of producing a sensitive and specific antigen diagnostic test for onchocerciasis.
...
PMID:Onchocerca volvulus: monoclonal antibodies to immune complex-associated parasite antigens. 769 Jul 12
Infective larvae (L3) of Onchocerca
volvulus
were procured in Liberia, West Africa, in the natural black fly vector, Simulium yahense. A cryobiological technique was developed to preserve L3 of O.
volvulus
that were fully viable after thawing. Larvae were treated before cooling with 4 cryoprotective compounds. Three compounds, dimethyl sulfoxide (DMSO), glycerol, and
ethylene glycol
, were prepared with distilled water. The fourth compound was DMSO prepared in different concentrations with 0.25 M sucrose. The treatment with DMSO + 0.25 M sucrose cryoprotectant resulted in the highest survival of infective larvae. Five cooling rates between 0.5 C/min and 20.0 C/min were applied. The highest survival of L3 was with the cooling rate of 1.0 C/min. Two-step cooling of L3 was applied. In the first step, L3's were frozen to 5 levels from -10.0 C to -20.0 C, -30.0 C, -40.0 C, -60 C, and -80.0 C, and in the second step, larvae were transferred into liquid nitrogen at -196 C for rapid cooling and storage. The survival was the highest when larvae were cooled to approximately -40 C prior to transfer into liquid nitrogen. Slow, gradual, and rapid thawing procedures were applied. The survival was the highest in rapid warming.
...
PMID:Cryopreservation of infective larvae of Onchocerca volvulus (Filarioidea: Onchocercidae). 841 May 41
Filarial parasites release macromolecules into their environment both in vitro and in vivo. These excretory-secretory products (E-S) have been studied with respect to function, vaccination potential, pathogenicity, and ability to serve as antigen targets for diagnostic tests. We have recently described monoclonal antibody OV-1 which binds to an intermediate filament in E-S and circulating antigens of Onchocerca
volvulus
. OV-1 also binds to cross-reactive antigens of Brugia malayi. Therefore, OV-1 was used to immunoscreen a B. malayi adult worm cDNA library in an attempt to clone a homologue (BMIF). BMIF is a 1664-bp full-length transcript which codes for 505 amino acids. BMIF has 95% sequence homology at the amino-acid level to OV1CF, an O.
volvulus
intermediate filament that was also selected with OV-1, and 75% homology to Ascaris intermediate filament A. Southern blot analysis suggests that BMIF is confined to a single location in the genomic DNA of B. malayi. Antibodies raised to BMIF identified native antigens in immunoblots of B. malayi adult worms, infective larvae and adult E-S. In addition, the antibody also bound to a 60-kDa antigen in immunoblots of poly(
ethylene glycol
)-precipitated immune complexes in sera from B. malayi infected patients. Localization studies showed that the antigen encoded by BMIF is present in the hypodermis, developing embryos and muscle of adult B. malayi. These studies show that BMIF is an E-S product of B. malayi adult worms which is detectable in sera from patients with brugian filariasis.
...
PMID:Molecular characterization of a Brugia malayi intermediate filament protein which is an excretory-secretory product of adult worms. 857 31
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