scholarly journals Evaluation of a Western Blot Test in an Outbreak of Acute Pulmonary Histoplasmosis

1999 ◽  
Vol 6 (1) ◽  
pp. 20-23 ◽  
Author(s):  
Claudia V. Pizzini ◽  
Rosely M. Zancopé-Oliveira ◽  
Errol Reiss ◽  
Rana Hajjeh ◽  
Leo Kaufman ◽  
...  
Keyword(s):  
Western Blot ◽  
Acute Phase ◽  
Periodate Oxidation ◽  
Negative Control ◽  
Acute Stage ◽  
Serum Samples ◽  
Specific Test ◽  
Carbohydrate Epitopes ◽  
Pulmonary Histoplasmosis ◽  
Control Serum

ABSTRACT A western blot (WB) test was evaluated for detection of antibodies against native glycosylated and chemically deglycosylated M and H antigens of Histoplasma capsulatum in serum obtained from patients during the acute phase of pulmonary histoplasmosis that occurred during an outbreak. Of 275 serum samples tested by immunodiffusion and complement fixation (CF) samples from 40 patients affected during this outbreak and from 37 negative controls were tested by WB test. A group of patients whose sera were negative for CF antibodies and precipitins early in the acute stage of histoplasmosis but who all seroconverted during convalescence 6 weeks later were tested with the WB test. Antibodies against untreated H and M antigens were detected at a 1:100 dilution by WB test in 45% of the 20 acute-phase serum samples and in all 20 of the convalescent-phase specimens. The WB test’s sensitivity for acute-phase specimens increased to 90% (18 of 20 specimens) when H and M antigens were treated by periodate oxidation to inactivate susceptible carbohydrate epitopes. When native glycosylated antigens were used in the WB test, positive reactions were observed in negative control serum specimens (3 of 37 specimens; 8%) and in serum specimens obtained from asymptomatic persons screened as part of the outbreak investigation (13 of 20 specimens; 65%). These positive reactions were also attributed to glycosidic epitopes since the specificity of the WB test increased from 78 to 100% when periodate-treated H and M antigens were used. WB test with deglycosylated H and M antigens of histoplasmin provides a rapid, sensitive, and specific test to diagnose acute pulmonary histoplasmosis before precipitins can be detected.

scholarly journals Dark Immunofluorescence: Correlation with Serum Immunoglobulin Abnormalities

2006 ◽  
Vol 13 (11) ◽  
pp. 1294-1295
Author(s):  
J. List ◽  
M. S. Buckland ◽  
B. Thobhani ◽  
C. J. Sheed ◽  
J. C. Mann ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Retrospective Review ◽  
Negative Control ◽  
Serum Immunoglobulin ◽  
Serum Samples ◽  
Serum Immunoglobulins ◽  
Control Serum ◽  
Negative Control Serum

ABSTRACT Occasional serum samples (<0.5%) tested by indirect immunofluorescence showed less fluorescence than did negative-control serum. A retrospective review of these patients' serum immunoglobulins revealed a high percentage of abnormalities (71%, versus 22% of controls). We suggest that this observation should be reported when seen and that the clinician should be alerted to an association with immunoglobulin abnormalities.

scholarly journals Natural Autoimmunity to the Thyroid Hormone Monocarboxylate Transporters MCT8 and MCT10

Biomedicines ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 496
Author(s):  
Theresa Porst ◽  
Jörg Johannes ◽  
Hans Gluschke ◽  
Richard Köhler ◽  
Sebastian Mehl ◽  
...  
Keyword(s):  
Cell Culture ◽  
Thyroid Hormone ◽  
Thyroid Disease ◽  
Biological Effects ◽  
Negative Control ◽  
Monocarboxylate Transporters ◽  
Healthy Controls ◽  
Serum Samples ◽  
Control Serum ◽  
Overweight Adolescents

The monocarboxylate transporters 8 (MCT8) and 10 (MCT10) are important for thyroid hormone (TH) uptake and signaling. Reduced TH activity is associated with impaired development, weight gain and discomfort. We hypothesized that autoantibodies (aAb) to MCT8 or MCT10 are prevalent in thyroid disease and obesity. Analytical tests for MCT8-aAb and MCT10-aAb were developed and characterized with commercial antiserum. Serum samples from healthy controls, thyroid patients and young overweight subjects were analyzed, and prevalence of the aAb was compared. MCT8-aAb were additionally tested for biological effects on thyroid hormone uptake in cell culture. Positive MCT8-aAb and MCT10-aAb were detected in all three clinical cohorts analyzed. MCT8-aAb were most prevalent in thyroid patients (11.9%) as compared to healthy controls (3.8%) and overweight adolescents (4.2%). MCT8-aAb positive serum reduced T4 uptake in cell culture in comparison to MCT8-aAb negative control serum. Prevalence of MCT10-aAb was highest in the group of thyroid patients as compared to healthy subjects or overweight adolescents (9.0% versus 4.5% and 6.3%, respectively). We conclude that MCT8 and MCT10 represent autoantigens in humans, and that MCT8-aAb may interfere with regular TH uptake and signaling. The increased prevalence of MCT8-aAb and MCT10-aAb in thyroid disease suggests that their presence may be of pathophysiological relevance. This hypothesis deserves an analysis in large prospective studies.

scholarly journals Serological signatures of SARS-CoV-2 infection: Implications for antibody-based diagnostics

2020 ◽  
Author(s):  
Jason Rosado ◽  
Stéphane Pelleau ◽  
Charlotte Cockram ◽  
Sarah Hélène Merkling ◽  
Narimane Nekkab ◽  
...  
Keyword(s):  
Population Level ◽  
Classification Performance ◽  
Negative Control ◽  
Antibody Responses ◽  
Serum Samples ◽  
Igg Antibody ◽  
Low Transmission ◽  
Control Serum ◽  
Challenges And Opportunities ◽  
Serological Diagnostics

AbstractBackgroundInfection with SARS-CoV-2 induces an antibody response targeting multiple antigens that changes over time. This complexity presents challenges and opportunities for serological diagnostics.MethodsA multiplex serological assay was developed to measure IgG and IgM antibody responses to seven SARS-CoV-2 spike or nucleoprotein antigens, two antigens for the nucleoproteins of the 229E and NL63 seasonal coronaviruses, and three non-coronavirus antigens. Antibodies were measured in serum samples from patients in French hospitals with RT-qPCR confirmed SARS-CoV-2 infection (n = 259), and negative control serum samples collected before the start of the SARS-CoV-2 epidemic (n = 335). A random forests algorithm was trained with the multiplex data to classify individuals with previous SARS-CoV-2 infection. A mathematical model of antibody kinetics informed by prior information from other coronaviruses was used to estimate time-varying antibody responses and assess the potential sensitivity and classification performance of serological diagnostics during the first year following symptom onset. A statistical estimator is presented that can provide estimates of seroprevalence in very low transmission settings.ResultsIgG antibody responses to trimeric Spike protein identified individuals with previous RT-qPCR confirmed SARS-CoV-2 infection with 91.6% sensitivity (95% confidence interval (CI); 87.5%, 94.5%) and 99.1% specificity (95% CI; 97.4%, 99.7%). Using a serological signature of IgG and IgM to multiple antigens, it was possible to identify infected individuals with 98.8% sensitivity (95% CI; 96.5%, 99.6%) and 99.3% specificity (95% CI; 97.6%, 99.8%). Informed by prior data from other coronaviruses, we estimate that one year following infection a monoplex assay with optimal anti-Stri IgG cutoff has 88.7% sensitivity (95% CI: 63.4%, 97.4%), and that a multiplex assay can increase sensitivity to 96.4% (95% CI: 80.9%, 100.0%). When applied to population-level serological surveys, statistical analysis of multiplex data allows estimation of seroprevalence levels less than 1%, below the false positivity rate of many other assays.ConclusionSerological signatures based on antibody responses to multiple antigens can provide accurate and robust serological classification of individuals with previous SARS-CoV-2 infection. This provides potential solutions to two pressing challenges for SARS-CoV-2 serological surveillance: classifying individuals who were infected greater than six months ago, and measuring seroprevalence in serological surveys in very low transmission settings.

Improved specificity of serum albumin determination and estimation of "acute phase reactants" by use of the bromcresol green reaction.

1976 ◽  
Vol 22 (5) ◽  
pp. 616-622 ◽  
Author(s):  
J E Gustafsson
Keyword(s):  
Serum Albumin ◽  
Cellulose Acetate ◽  
Acute Phase ◽  
Acute Phase Reactant ◽  
Serum Samples ◽  
Acute Phase Reactants ◽  
Slow Reaction ◽  
Phase Reactant ◽  
Control Serum ◽  
Bromcresol Green

Abstract The reaction of serum samples with bromcresol green proceeds in two steps. Albumin is responsible for the faster (less than 1 min) reaction; the slower (30-min) reaction is a measure of "acute phase reactant(s)" in serum. Serum is simply mixed with bromcresol green reagent and the absorbance is measured twice, immediately and at 60 min. Albumin concentrations, determined from the absorbance at 0 min, correlate well with those determined by Laurell "rocket" immunoelectrophoresis; r = 0.95 with no certain deviation from unity for the slope and with a negligible difference at zero concentration. The slow reaction was expressed as deltaA% = 100 (deltaAs/deltaAv) where deltaAs and delta Av are the changes in absorbance between 60 and 0 min for the sample and a commercial control serum, respectively. The value for deltaA% correlates well with the percentage of alpha2-fraction as determined by electrophoresis on cellulose acetate, as well as with orosomucoid and ceruloplasmin, all of which are acute phase reactant(s). Whether these proteins or other acute phase reactant(s) actually cause the slow reaction has not yet been established.

scholarly journals Natural Autoimmunity to the Thyroid Hormone Monocarboxylate Transporters MCT8 and MCT10

2021 ◽  
Author(s):  
Theresa Porst ◽  
Jörg Johannes ◽  
Hans Gluschke ◽  
Richard Köhler ◽  
Sebastian Mehl ◽  
...  
Keyword(s):  
Cell Culture ◽  
Thyroid Hormone ◽  
Thyroid Disease ◽  
Biological Effects ◽  
Negative Control ◽  
Monocarboxylate Transporters ◽  
Healthy Controls ◽  
Serum Samples ◽  
Control Serum ◽  
Overweight Adolescents

The monocarboxylate transporters 8 (MCT8) and 10 (MCT10) are important for thyroid hormone (TH) uptake and signaling. Reduced TH activity is associated with impaired development, weight gain and discomfort. We hypothesized that autoantibodies (aAb) to MCT8 or MCT10 are prevalent in thyroid disease and obesity. Analytical tests for MCT8-aAb and MCT10-aAb were developed and characterized with commercial antiserum. Serum samples from healthy controls, thyroid patients and young overweight subjects were analyzed, and prevalence of the aAb was compared. MCT8-aAb were additionally tested for biological effects on thyroid hormone uptake in cell culture. Positive MCT8-aAb and MCT10-aAb were detected in all three clinical cohorts analyzed. MCT8-aAb were most prevalent in thyroid patients (11.9%) as compared to healthy controls (3.8%) and overweight adolescents (4.2%). MCT8-aAb positive serum reduced T4 uptake in cell culture in comparison to MCT8-aAb negative control serum. Prevalence of MCT10-aAb was highest in the group of thyroid patients as compared to healthy subjects or overweight adolescents (9.0% versus 4.5% and 6.3%, respectively). We conclude that MCT8 and MCT10 represent autoantigens in humans, and that MCT8-aAb may interfere with regular TH uptake and signaling. The increased prevalence of MCT8-aAb and MCT10-aAb in thyroid disease suggests that their presence may be of pathophysiological relevance. This hypothesis deserves an analysis in large prospective studies.

scholarly journals 233 NaturSafe® supplementation mitigates some aspects of the acute phase immune response to a lipopolysaccharide (LPS) challenge in weaned beef calves

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 117-117
Author(s):  
Nicole C Burdick Sanchez ◽  
Jeffery A Carroll ◽  
Paul R Broadway ◽  
Tom S Edrington ◽  
Ilkyu Yoon ◽  
...  
Keyword(s):  
Acute Phase ◽  
Treatment Time ◽  
Sickness Behavior ◽  
Serum Cortisol ◽  
Post Challenge ◽  
Serum Samples ◽  
Lps Challenge ◽  
Time Interaction ◽  
Tnf Α ◽  
Monitoring Devices

Abstract This study was conducted to determine if feeding calves NaturSafe would reduce the acute phase response (APR) to lipopolysaccharide (LPS) challenge. Crossbred steers (n=32; 274±2 kg) were randomly allotted to two treatment diets: 1) Control, fed a standard receiving ration, and 2) NaturSafe, fed the Control ration supplemented with NaturSafe at 12 g/hd/d (NaturSafe®, Diamond V). On d22, steers were fitted with indwelling jugular catheters and rectal temperature monitoring devices and placed in individual stalls. On d23, steers were challenged i.v. with 0.25 µg/kg BW LPS. Serum samples were collected and sickness behavior scores (SBS) recorded at 0.5-h intervals from -2 to 8h and at 24h relative to LPS challenge. Rectal temperatures were greater (P=0.01) in NaturSafe compared to Control steers for the following time intervals following LPS challenge: 6 to 11h, 13h, 15 to 20h, and 22 to 24h. Additionally, SBS were reduced (P&lt; 0.01) in NaturSafe compared to Control steers. White blood cell concentrations were greater (P=0.05) in NaturSafe compared to Control steers prior to the LPS challenge, yet the response to LPS did not differ between treatments (P &gt;0.05). A treatment × time interaction for serum cortisol concentrations (P&lt; 0.01) showed an increase at 0.5 and 2h post-challenge but a reduction at 3h in NaturSafe compared to Control steers. Additionally, fibrinogen was greater (P&lt; 0.01) in NaturSafe compared to Control steers. There was a treatment × time interaction (P&lt; 0.01) for TNF-α where concentrations were reduced from 1 to 2h post-challenge in NaturSafe compared to Control steers. Serum IL-6 tended (P=0.09) to show a reduction in serum concentrations in NaturSafe compared to Control steers. There was a tendency (P=0.07) for a treatment × time interaction for IFN-γ. Overall these data suggest a priming effect of NaturSafe on the innate immune system of steers, resulting in an attenuated APR to the LPS challenge.

scholarly journals Non-Invasive Dengue Diagnostics—The Use of Saliva and Urine for Different Stages of the Illness

Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1345
Author(s):  
Mahathir Humaidi ◽  
Wei Ping Tien ◽  
Grace Yap ◽  
Choon Rong Chua ◽  
Lee Ching Ng
Keyword(s):  
Clinical Symptoms ◽  
Detection Algorithm ◽  
Acute Stage ◽  
Serum Samples ◽  
Non Invasive ◽  
Fever Onset ◽  
Dengue Transmission ◽  
Dengue Diagnostics ◽  
Potential Tool

Dengue diagnosis is largely dependent on clinical symptoms and routinely confirmed with laboratory detection of dengue virus in patient serum samples collected via phlebotomy. This presents a challenge to patients not amenable to venipuncture. Non-invasive methods of dengue diagnosis have the potential to enhance the current dengue detection algorithm. In this study, samples from dengue infected patients were collected between January 2012 until September 2012 and September 2013 until December 2013 in two different setups. Panel A samples (blood, urine, and saliva) were collected daily when the 39 patients were hospitalised and during their follow-up visits while Panel B samples (saliva) were collected from 23 patients during the acute stage of dengue. Using DENV PCR on Panel A, from day 2 to day 4 post fever onset, serum showed the best overall positivity followed by saliva and urine (100%/82.1%/67.9%). From day 5 until day 10 post fever onset, serum and urine had similar positivity (67.4%/61.2%), followed by saliva (51.3%). Beyond day 10 post fever onset, DENV was undetectable in sera, but urine and saliva showed 56.8% and 28.6% positivity, respectively. DENV in urine was detectable up until 32 days post fever. Panel B results showed overall sensitivity of 32.4%/36% (RNA/NS1) for DENV detection in saliva. Our results suggest that the urine-based detection method is useful especially for late dengue detection, where DENV is undetected in sera but still detectable in urine. This provides a potential tool for the physician to pick up new cases in an area where there is ongoing dengue transmission and subsequently prompt for intensified vector control activities.

scholarly journals Physiological parameters deterioration in acute stage after stroke: Rehabilitation versus conventional care

2019 ◽  
Vol 10 (3) ◽  
pp. 79
Author(s):  
Reham AbdElhamed AbdElmawla Elsaid ◽  
Amina Mohamed AbdElfatah Sliman
Keyword(s):  
Acute Stroke ◽  
Acute Phase ◽  
Stroke Rehabilitation ◽  
Recovery Period ◽  
Acute Stage ◽  
Conventional Care ◽  
Physiological Parameters ◽  
Stroke Recovery ◽  
University Hospital ◽  
Control Group

Objective: Stroke is considered the main health problem and the second leading cause of death worldwide. Stroke resulting in varied and unpredictable complications if not managed correctly in the acute stage with intensive rehabilitation therapy which may affect stroke prognosis, and resulting functional decline. Therefore, the aim of the study was to explore the consequences of rehabilitation versus conventional care on physiological parameters during the acute stroke recovery period.Methods: The quasi-experimental research design was used in the neurology department at Mansoura University Hospital. A convenient sample of sixty-four adult patients of both sex with stroke, who corresponded to inclusion criteria was assigned into two equal groups, study group (rehabilitation group) and control group (conventional care).Results: The results indicates, acute phase rehabilitation limit physiological parameters deterioration during acute stroke recovery period comparing to conventional care only.Conclusions: Acute phase stroke rehabilitation has a significant positive impact on physiological parameters.

scholarly journals Rabbit and rat C-reactive proteins bind apolipoprotein B-containing lipoproteins.

1984 ◽  
Vol 159 (2) ◽  
pp. 604-616 ◽  
Author(s):  
I F Rowe ◽  
A K Soutar ◽  
I M Trayner ◽  
M L Baltz ◽  
F C de Beer ◽  
...  
Keyword(s):  
Acute Phase ◽  
Apolipoprotein B ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Plasma Lipoproteins ◽  
Free Form ◽  
Low Density ◽  
Serum Samples ◽  
Acute Phase Serum

Immobilized rabbit and rat C-reactive protein (CRP) were found to selectively bind apolipoprotein B (apoB)-containing lipoproteins (low density lipoprotein, LDL and very low density lipoprotein, VLDL) from whole serum in a manner similar to that previously reported with human CRP. In acute phase human serum the CRP is in a free form, not complexed with lipoprotein or any other macromolecular ligand, and in acute phase serum from most rabbits fed on a normal diet the rabbit CRP was also free. However, in acute phase serum or heparinized plasma from hypercholesterolemic rabbits part or all of the CRP was found by gel filtration and immunoelectrophoretic techniques to be complexed with beta-VLDL, an abnormal apoB-containing plasma lipoprotein present in these animals. The presence of extent in different serum samples of CRP complexed with lipoprotein correlated closely with the serum apoB concentration. The formation of complexes between native, unaggregated rabbit CRP in solution and apoB-containing lipoproteins was readily demonstrable experimentally both with the isolated proteins and in whole serum. In all cases these interactions were calcium-dependent and inhibitable by free phosphoryl choline. The present findings extend earlier work in man and the rabbit and indicate that among the C-reactive proteins from different species, which are structurally highly conserved, the capacity for selective binding to apoB-containing plasma lipoproteins is also a constant feature. These interactions may therefore be related to the in vivo function of CRP in all species and this function may in turn be relevant to pathological conditions, such as atherosclerosis, in which lipoproteins are important.

α-Tocopherol, lipids and lipoproteins in knee-joint synovial fluid and serum from patients with inflammatory joint disease

1992 ◽  
Vol 83 (6) ◽  
pp. 657-664 ◽  
Author(s):  
K. Fairburn ◽  
M. Grootveld ◽  
R. J. Ward ◽  
C. Abiuka ◽  
M. Kus ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Serum Levels ◽  
Biologically Active ◽  
Joint Disease ◽  
Inflammatory Joint Disease ◽  
Serum Samples ◽  
Control Serum ◽  
Inflammatory Synovial Fluid

1. We have determined the antioxidant status of synovial fluid and serum of patients with inflammatory joint disease in terms of the biologically active lipid-soluble antioxidant, α-tocopherol. Synovial fluid concentrations of α-tocopherol were significantly lower relative to those of paired serum samples (P<0.001). Serum levels of α-tocopherol in these patients did not differ significantly from those in control serum. 2. Lower concentrations of cholesterol, triacylglycerol and low-density lipoprotein were also observed in patients' synovial fluid compared with matched serum samples. However, multiple regression analysis of the data indicated that there remained a significant depletion of α-tocopherol, which was largely independent of these co-variables, in inflammatory synovial fluid. These findings are consistent with the consumption of α-tocopherol within the inflamed joint via its role in terminating the process of lipid peroxidation. 3. Nuclear magnetic resonance spectroscopic analysis of matched inflammatory synovial fluid and serum confirmed lower concentrations of triacylglycerol in synovial fluid together with evidence of a shortened mean triacylglycerol chain length. The latter metabolic difference suggests an increased utilization of triacylglycerols for energy within the inflamed joint.

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