Journal of Clinical Medicine

Research

18 pages, 2072 KiB

Open AccessArticle

Environmental Enrichment Components Required to Reduce Methamphetamine-Induced Behavioral Sensitization in Mice: Examination of Behaviors and Neural Substrates

by Cai-N Cheng, Shaw-Jye Wu and Andrew Chih Wei Huang

J. Clin. Med. 2022, 11(11), 3051; https://doi.org/10.3390/jcm11113051 - 28 May 2022

Cited by 2 | Viewed by 2166

Abstract

Environmental enrichment (EE) involves the presentation of various sensory, physical, social, and cognitive stimuli in order to alter neural activity in specific brain areas, which can ameliorate methamphetamine (MAMPH)-induced behavioral sensitization and comorbid anxiety symptoms. No previous studies have comprehensively examined which EE [...] Read more.

Environmental enrichment (EE) involves the presentation of various sensory, physical, social, and cognitive stimuli in order to alter neural activity in specific brain areas, which can ameliorate methamphetamine (MAMPH)-induced behavioral sensitization and comorbid anxiety symptoms. No previous studies have comprehensively examined which EE components are critical for effectively reducing MAMPH-induced behavioral sensitization and anxiety. This study examined different housing conditions, including standard housing (SH, No EE), standard EE (STEE), physical EE (PEE), cognitive EE (CEE), and social EE (SEE). In the beginning, mice were randomly assigned to the different combinations of housing conditions and injections, consisting of No EE/Saline, No EE/MAMPH, STEE/MAMPH, PEE/MAMPH, CEE/MAMPH, and SEE/MAMPH groups. Then, the mice received intraperitoneal injections of 1 mg/kg MAMPH or normal saline daily for 7 days, followed by a final injection of 0.5 mg/kg MAMPH or normal saline. After behavioral tests, all mice were examined for c-Fos immunohistochemical staining. The results showed that MAMPH induced behavioral sensitization as measured by distance traveled. MAMPH appeared to induce lowered anxiety responses and severe hyperactivity. All EE conditions did not affect MAMPH-induced lowered anxiety behaviors. STEE was likely more effective for reducing MAMPH-induced behavioral sensitization than PEE, CEE, and SEE. The c-Fos expression analysis showed that the medial prefrontal cortex (i.e., cingulate cortex 1 (Cg1), prelimbic cortex (PrL), and infralimbic cortex (IL)), nucleus accumbens (NAc), basolateral amygdala (BLA), ventral tegmental area (VTA), caudate-putamen (CPu), and hippocampus (i.e., CA1, CA3, and dentate gyrus (DG)) contributed to MAMPH-induced behavioral sensitization. The Cg1, IL, NAc, BLA, VTA, CPu, CA3, and DG also mediated STEE reductions in MAMPH-induced behavioral sensitization. This study indicates that all components of EE are crucial for ameliorating MAMPH-induced behavioral sensitization, as no individual EE component was able to effectively reduce MAMPH-induced behavioral sensitization. The present findings provide insight into the development of non-pharmacological interventions for reducing MAMPH-induced behavioral sensitization. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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13 pages, 1424 KiB

Open AccessArticle

Assessing Prevalence and Transmission Rates of Malaria through Simultaneous Profiling of Antibody Responses against Plasmodium and Anopheles Antigens

by Sidhartha Chaudhury, Jessica S. Bolton, Leigh Anne Eller, Merlin Robb, Julie Ake, Viseth Ngauy, Jason A. Regules, Edwin Kamau and Elke S. Bergmann-Leitner

J. Clin. Med. 2022, 11(7), 1839; https://doi.org/10.3390/jcm11071839 - 26 Mar 2022

Cited by 2 | Viewed by 2307

Abstract

Reliably assessing exposure to mosquitoes carrying malaria parasites continues to be a challenge due to the lack of reliable, highly sensitive diagnostics with high-throughput potential. Here, we describe an approach that meets these requirements by simultaneously measuring immune responses to both disease vector [...] Read more.

Reliably assessing exposure to mosquitoes carrying malaria parasites continues to be a challenge due to the lack of reliable, highly sensitive diagnostics with high-throughput potential. Here, we describe an approach that meets these requirements by simultaneously measuring immune responses to both disease vector and pathogen, using an electro-chemiluminescence-based multiplex assay platform. While using the same logistical steps as a classic ELISA, this platform allows for the multiplexing of up to ten antigens in a single well. This simple, reproducible, quantitative readout reports the magnitude, incidence, and prevalence of malaria infections in residents of malaria-endemic areas. By reporting exposure to both insect vectors and pathogen, the approach also provides insights into the efficacy of drugs and/or other countermeasures deployed against insect vectors aimed at reducing or eliminating arthropod-borne diseases. The high throughput of the assay enables the quick and efficient screening of sera from individuals for exposure to Plasmodium even if they are taking drug prophylaxis. We applied this assay to samples collected from controlled malaria infection studies, as well as those collected in field studies in malaria-endemic regions in Uganda and Kenya. The assay was sensitive to vector exposure, malaria infection, and endemicity, demonstrating its potential for use in malaria serosurveillance. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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13 pages, 908 KiB

Open AccessArticle

Oxidative and Antioxidative Status Expressed as OSI Index and GSH/GSSG Ratio in Children with Bone Tumors after Anticancer Therapy Completion

by Joanna Gajewska, Magdalena Chełchowska, Magdalena Rychłowska-Pruszyńska, Teresa Klepacka and Jadwiga Ambroszkiewicz

J. Clin. Med. 2022, 11(6), 1663; https://doi.org/10.3390/jcm11061663 - 17 Mar 2022

Cited by 5 | Viewed by 2166

Abstract

Aims. There are no data on the redox status of children with bone tumors in complete disease remission. Therefore, the presented study examined the reduced/oxidized glutathione (GSH/GSSG) ratio, total oxidant capacity (TOC) and total antioxidant capacity (TAC) values as well as the oxidative [...] Read more.

Aims. There are no data on the redox status of children with bone tumors in complete disease remission. Therefore, the presented study examined the reduced/oxidized glutathione (GSH/GSSG) ratio, total oxidant capacity (TOC) and total antioxidant capacity (TAC) values as well as the oxidative stress index (OSI) for assessing alterations in the oxidant/antioxidant balance in 35 children with osteosarcoma or Ewing’s sarcoma after anticancer therapy completion (median 14 months) compared with a control group. Methods. GSH, GSSG, TOC, TAC concentrations and bone alkaline phosphatase (BALP) activity were evaluated by immunoenzymatic (ELISA) and enzymatic methods. Results. We found no differences in serum BALP activity between all survivors with bone tumors and the control group. Patients with osteosarcoma after anticancer therapy completion had significantly higher values of TAC, GSH and the GSH/GSSG ratio as well as GSSG than healthy subjects. In patients with Ewing’s sarcoma, we found significantly higher values of TOC concentration compared with healthy children. In addition, survivors with Ewing’s sarcoma had higher TOC concentrations and OSI index values (p < 0.01), but a lower GSH/GSSG ratio (p < 0.05) than survivors with osteosarcoma. A positive correlation between TOC and the post-therapy period was observed in survivors. Conclusions. We found that in survivors with bone tumors, a disturbed balance between prooxidants and antioxidants persists after the completion of anticancer treatment. Moreover, an increased TOC value together with the post-therapy period may suggest increasing oxidative processes in survivors with bone tumors after treatment. Further observations will allow assessment of the relationship between the oxidant/antioxidant status and the predisposition of survivors to bone neoplastic disease recurrence. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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17 pages, 1390 KiB

Open AccessArticle

Decreased Serum Levels of SIRT1 and SIRT3 Correlate with Severity of Skin and Lung Fibrosis and Peripheral Microvasculopathy in Systemic Sclerosis

by Mirko Manetti, Irene Rosa, Bianca Saveria Fioretto, Marco Matucci-Cerinic and Eloisa Romano

J. Clin. Med. 2022, 11(5), 1362; https://doi.org/10.3390/jcm11051362 - 1 Mar 2022

Cited by 15 | Viewed by 3634

Abstract

Systemic sclerosis (SSc, scleroderma) is a severe autoimmune connective tissue disease characterized by widespread peripheral microvasculopathy, and progressive cutaneous and visceral fibrosis, leading to significant organ dysfunction. Sirtuins (SIRTs) are a family of NAD-dependent protein deacetylases with pleiotropic effects on a variety of [...] Read more.

Systemic sclerosis (SSc, scleroderma) is a severe autoimmune connective tissue disease characterized by widespread peripheral microvasculopathy, and progressive cutaneous and visceral fibrosis, leading to significant organ dysfunction. Sirtuins (SIRTs) are a family of NAD-dependent protein deacetylases with pleiotropic effects on a variety of biological processes, including metabolism, cell survival, and aging. In the last decades, increasing studies have explored the contribution of SIRTs to the pathogenesis of SSc, highlighting a significant antifibrotic effect of both SIRT1 and SIRT3. On these bases, the aim of this study was to measure circulating SIRT1 and SIRT3 levels by enzyme-linked immune-sorbent assay in a well-characterized cohort of SSc patients (n = 80) and healthy controls (n = 71), focusing on their possible association with disease clinical features, and their potential as biomarkers reflecting SSc activity and severity. Significantly decreased serum levels of both SIRT1 and SIRT3 were found in SSc patients compared to controls. In SSc, the reduction in circulating SIRT1 and SIRT3 associated with a greater extent of cutaneous fibrosis, presence of interstitial lung disease, and worse pulmonary function. Serum SIRT1 and SIRT3 decrease also correlated with the severity of nailfold microvascular damage, with SIRT3 levels being additionally related to the occurrence of digital ulcers. The levels of these two proteins showed a direct correlation with one another in the circulation of SSc patients. Of the two SIRTs, serum SIRT3 was found to better reflect disease activity and severity in a logistic regression analysis model. Our findings suggest that serum SIRT1 and SIRT3 may represent novel potential biomarkers of increased risk for a more severe, life-threatening SSc disease course. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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10 pages, 1056 KiB

Open AccessArticle

Is Oxidative Stress an Emerging Player in the Thrombosis of Patients with Anti-Phosphatidylethanolamine Autoantibodies?

by Xavier Heim, Daniel Bertin, Noémie Resseguier, Abdelouahab Beziane, Audrey Metral, Alexandre Brodovitch, Régis Guieu, Jean-Guillaume Steinberg, Marcel Blot-Chabaud, Pierre-Emmanuel Morange, Jean-Louis Mege and Nathalie Bardin

J. Clin. Med. 2022, 11(5), 1297; https://doi.org/10.3390/jcm11051297 - 27 Feb 2022

Cited by 2 | Viewed by 1716

Abstract

The detection of anti-phosphatidylethanolamine autoantibodies (aPEs) has been proposed to improve the diagnosis and management of patients presenting clinical manifestations of antiphospholipid syndrome (APS), such as thrombosis, and who are persistently negative for conventional markers. After selecting the most specific ELISA for their [...] Read more.

The detection of anti-phosphatidylethanolamine autoantibodies (aPEs) has been proposed to improve the diagnosis and management of patients presenting clinical manifestations of antiphospholipid syndrome (APS), such as thrombosis, and who are persistently negative for conventional markers. After selecting the most specific ELISA for their detection, we evidenced the interest of aPEs in the exploration of thrombosis when APS conventional markers were negative through a 1-year retrospective study including 1131 consecutive patients routinely tested for aPEs. To validate this result, we assessed aPEs in a newly selected population of 77 patients with unexplained deep vein thrombosis (DVT). With a total prevalence of 19.5%, we confirmed the interest of aPE detection in patients with unexplained DVT who were devoid of other aPLs markers. Since endosomal compartment, a source of ROS production, has been recently identified as the cellular target of aPEs in vitro, we then investigated an association between aPE positivity and reactive oxygen species (ROS) production by measuring the production of thiobarbituric acid-reactive substances. We showed, for the first time, a significant association between aPE positivity and systemic ROS production in patients which led us to hypothesize a new mechanism of action of aPEs in thrombosis through a signaling related to oxidative stress. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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12 pages, 3800 KiB

Open AccessArticle

A Microplate-Based Approach to Map Interactions between TDP-43 and α-Synuclein

by Angelo M. Jamerlan and Seong Soo A. An

J. Clin. Med. 2022, 11(3), 573; https://doi.org/10.3390/jcm11030573 - 24 Jan 2022

Cited by 2 | Viewed by 3616

Abstract

Trans-active response DNA-binding protein (TDP-43) is a multifunctional regulatory protein, whose abnormal deposition in neurons was linked to debilitating neurodegenerative diseases, such as amyotrophic lateral sclerosis, frontotemporal lobar degeneration, Limbic-predominant age-related TDP-43 encephalopathy, and Alzheimer’s disease with a secondary pathology. Several reports showed [...] Read more.

Trans-active response DNA-binding protein (TDP-43) is a multifunctional regulatory protein, whose abnormal deposition in neurons was linked to debilitating neurodegenerative diseases, such as amyotrophic lateral sclerosis, frontotemporal lobar degeneration, Limbic-predominant age-related TDP-43 encephalopathy, and Alzheimer’s disease with a secondary pathology. Several reports showed that TDP-43 proteinopathy as a comorbidity can form aggregates with other pathological proteins. The co-deposition of alpha synuclein and TDP-43 inclusions was previously reported in glial cells and by observing TDP-43 proteinopathy in Lewy body disease. In this study, it was hypothesized that alpha synuclein and TDP-43 may co-aggregate, resulting in comorbid synucleinopathy and TDP-43 proteinopathy. A solid-phase microplate-based immunoassay was used to map out the epitopes of anti-TDP-43 antibodies and locate the interaction of TDP-43 with α-synuclein. A region of the low complexity domain of TDP-43 (aa 311–314) was shown to interact with full-length α-synuclein. Conversely, full-length TDP-43 was shown to bind to the non-amyloid beta component of α-synuclein. Using in silico sequence-based prediction, the affinity and dissociation constant of full-length TDP-43 and α-synuclein were calculated to be −10.83 kcal/mol and 1.13 × 10⁻⁸, respectively. Taken together, this microplate-based method is convenient, economical, and rapid in locating antibody epitopes as well as interaction sites of two proteins. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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18 pages, 1970 KiB

Open AccessArticle

Freshwater Clam Extract Mitigates Neuroinflammation and Amplifies Neurotrophic Activity of Glia: Insights from In Vitro Model of Neurodegenerative Pathomechanism

by Muh-Shi Lin, Shu-Mei Chen, Kuo-Feng Hua, Wei-Jung Chen, Cho-Chen Hsieh and Chai-Ching Lin

J. Clin. Med. 2022, 11(3), 553; https://doi.org/10.3390/jcm11030553 - 22 Jan 2022

Viewed by 2596

Abstract

Background. An extensive body of research suggests that brain inflammation and oxidative stress are the underlying causes of Parkinson’s disease (PD), for which no potent therapeutic approach exists to mitigate the degradation of dopamine neurons. Freshwater clams, an ancient health food of Chinese [...] Read more.

Background. An extensive body of research suggests that brain inflammation and oxidative stress are the underlying causes of Parkinson’s disease (PD), for which no potent therapeutic approach exists to mitigate the degradation of dopamine neurons. Freshwater clams, an ancient health food of Chinese origin, have been documented to exhibit anti-inflammatory and antioxidant effects. We previously reported that freshwater clam extract (FCE) can attenuate astrocytic activation and subsequent proinflammatory cytokine production from substantia nigra in an MPTP-induced PD mouse model. This article provides insight into the potential mechanisms through which FCE regulates neuroinflammation in a glia model of injury. Materials and methods. In total, 1 μg/mL lipopolysaccharide (LPS) and 200 μM rotenone were conducted in primary glial cell cultures to mimic the respective neuroinflammation and oxidative stress during injury-induced glial cell reactivation, which is relevant to the pathological process of PD. Results. FCE markedly reduced LPS-induced neuroinflammation by suppressing NO and TNF-α production and the expression of pro-inflammatory cytokines. In addition, FCE was effective at reducing rotenone-induced toxicity by diminishing ROS production, promoting antioxidant enzymes (SOD, catalase, and GPx) and minimizing the decline in glial-cell-secreted neurotrophic factors (GDNF, BDNF). These impacts ultimately led to a decrease in glial apoptosis. Conclusions. Evidence reveals that FCE is capable of stabilizing reactive glia, as demonstrated by reduced neuroinflammation, oxidative stress, the increased release of neurotrophic factors and the inhibition of apoptosis, which provides therapeutic insight into neurodegenerative diseases, including PD. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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11 pages, 634 KiB

Open AccessArticle

ELISA-Based Analysis Reveals an Anti-SARS-CoV-2 Protein Immune Response Profile Associated with Disease Severity

by Charline Herrscher, Sébastien Eymieux, Christophe Gaborit, Hélène Blasco, Julien Marlet, Karl Stefic, Philippe Roingeard, Leslie Grammatico-Guillon and Christophe Hourioux

J. Clin. Med. 2022, 11(2), 405; https://doi.org/10.3390/jcm11020405 - 14 Jan 2022

Cited by 3 | Viewed by 1900

Abstract

Since the start of the COVID-19 pandemic, many studies have investigated the humoral response to SARS-CoV-2 during infection. Studies with native viral proteins constitute a first-line approach to assessing the overall immune response, but small peptides are an accurate and valuable tool for [...] Read more.

Since the start of the COVID-19 pandemic, many studies have investigated the humoral response to SARS-CoV-2 during infection. Studies with native viral proteins constitute a first-line approach to assessing the overall immune response, but small peptides are an accurate and valuable tool for the fine characterization of B-cell epitopes, despite the restriction of this approach to the determination of linear epitopes. In this study, we used ELISA and peptides covering a selection of structural and non-structural SARS-CoV-2 proteins to identify key epitopes eliciting a strong immune response that could serve as a biological signature of disease characteristics, such as severity, in particular. We used 213 plasma samples from a cohort of patients well-characterized clinically and biologically and followed for COVID-19 infection. We found that patients developing severe disease had higher titers of antibodies mapping to multiple specific epitopes than patients with mild to moderate disease. These data are potentially important as they could be used for immunological profiling to improve our knowledge of the quantitative and qualitative characteristics of the humoral response in relation to patient outcome. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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7 pages, 222 KiB

Open AccessArticle

Fecal Calprotectin Is Increased in Stroke

by Shin Young Park, Sang Pyung Lee and Woo Jin Kim

J. Clin. Med. 2022, 11(1), 159; https://doi.org/10.3390/jcm11010159 - 29 Dec 2021

Cited by 4 | Viewed by 1825

Abstract

Background: While there have been major advances in unveiling the mechanisms comprising the ischemic cascade of CNS, stroke continues to be a significant burden. There is a need to extend the focus toward peripheral changes, and the brain–gut axis has recently gained much [...] Read more.

Background: While there have been major advances in unveiling the mechanisms comprising the ischemic cascade of CNS, stroke continues to be a significant burden. There is a need to extend the focus toward peripheral changes, and the brain–gut axis has recently gained much attention. Our study aimed to evaluate gut inflammation and its association with blood variables in stroke using fecal calprotectin (FC). Methods: Fecal samples were obtained from 27 stroke patients and 27 control subjects. FC was quantitatively measured using a commercial ELISA. Laboratory data on the fecal sample collection were also collected, including CBC, ESR, glucose, creatinine, total protein, albumin, transaminases, and CRP. Results: There was a significant increase in FC levels in stroke patients compared to the controls. Furthermore, FC in stroke patients was negatively correlated with the Glasgow Coma Scale. Moreover, FC in stroke patients was positively correlated with CRP and negatively correlated with lymphocyte count and albumin. Conclusions: Our findings show that increased FC is associated with consciousness and systemic response in stroke and warrants further studies to elucidate the usefulness of FC in the management of stroke. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

12 pages, 601 KiB

Open AccessArticle

Association between Enzyme-Linked Immunosorbent Assay-Measured Kidney Injury Markers and Urinary Cadmium Levels in Chronic Kidney Disease

by Kai-Fan Tsai, Pai-Chin Hsu, Chien-Te Lee, Chia-Te Kung, Yi-Chin Chang, Lung-Ming Fu, Yu-Che Ou, Kuo-Chung Lan, Tzung-Hai Yen and Wen-Chin Lee

J. Clin. Med. 2022, 11(1), 156; https://doi.org/10.3390/jcm11010156 - 29 Dec 2021

Cited by 16 | Viewed by 2019

Abstract

Cadmium exposure is associated with chronic kidney disease (CKD), but the optimal biomarker for early cadmium-associated nephrotoxicity in low-level exposure has not yet been established. We conducted a cross-sectional investigation involving 167 CKD patients stratified according to tertiles of urinary cadmium levels (UCd), [...] Read more.

Cadmium exposure is associated with chronic kidney disease (CKD), but the optimal biomarker for early cadmium-associated nephrotoxicity in low-level exposure has not yet been established. We conducted a cross-sectional investigation involving 167 CKD patients stratified according to tertiles of urinary cadmium levels (UCd), in which enzyme-linked immunosorbent assay (ELISA)-measured novel renal biomarkers were utilized to assess the extent of renal injury associated with cadmium burden. In the analyses, urinary kidney injury molecule-1 (KIM-1) levels and age were the independent factors positively correlated with UCd after adjusting for covariates in non-dialysis-dependent CKD patients (high vs. low UCd, odds ratio (95% confidence interval), 1.0016 (1.0001–1.0032), p = 0.043, and 1.0534 (1.0091–1.0997), p = 0.018). Other conventional and novel renal biomarkers, such as serum creatinine, estimated glomerular filtration rate, CKD staging, urinary protein/creatinine ratio, urinary 8-hydroxy-2-deoxyguanosine (8-OHdG), and urinary epidermal growth factor (EGF) were not independently correlated with UCd in the analyses. In conclusion, our study found that the ELISA-measured urinary KIM-1 level could serve as an early renal injury marker in low-level cadmium exposure for non-dialysis-dependent CKD patients. In addition, age was an independent factor positively associated with UCd in this population. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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8 pages, 596 KiB

Open AccessCommunication

Vascular Endothelial Growth Factor as a Putative Biomarker of Depression in Asthmatics with Reversible Airway Narrowing

by Krzysztof Gomułka, Jerzy Liebhart and Wojciech Mędrala

J. Clin. Med. 2021, 10(22), 5301; https://doi.org/10.3390/jcm10225301 - 15 Nov 2021

Cited by 2 | Viewed by 1625

Abstract

The vascular endothelial growth factor (VEGF) plays a pivotal role in process of angiogenesis in adults. If angiogenesis is not properly controlled, its deregulation may implicate it in various psychosomatic diseases states. The aim of our study was to reveal possible correlation between [...] Read more.

The vascular endothelial growth factor (VEGF) plays a pivotal role in process of angiogenesis in adults. If angiogenesis is not properly controlled, its deregulation may implicate it in various psychosomatic diseases states. The aim of our study was to reveal possible correlation between severity of depression in asthmatics with different degrees of airway narrowing and serum vascular endothelial growth factor levels. The study population included a total of 122 adult subjects: 82 patients with asthma (among them 42 patients with irreversible bronchoconstriction and 40 patients with reversible bronchoconstriction) and 40 healthy participants as a control group. The standardized Beck Depression Inventory (BDI) was used to estimate the depression symptoms. Enzyme-linked immunosorbent assay (ELISA) was used to assess the VEGF serum concentration in all participants. There was a significant difference in depression symptoms in asthmatics with reversible (p = 0.0432) and irreversible airway obstruction (p = 0.00005) in comparison to control group and between these two subgroups of asthmatics (p = 0.0233). Obtained results revealed significant correlation between level of depression and mean VEGF serum concentration in asthmatics with reversible airway obstruction (p = 0.0202). There was no difference between enhanced depression symptoms and VEGF serum concentration in patients with irreversible airway obstruction nor in the total group of asthmatics (in both p > 0.05). The relationship between asthma severity and depression symptoms seems to be certain. VEGF might be considered as a putative biomarker of depression in asthmatics, mainly those with reversible airway narrowing. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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13 pages, 2295 KiB

Open AccessArticle

Non-Surgical Periodontal Therapy Could Improve the Periodontal Inflammatory Status in Patients with Periodontitis and Chronic Hepatitis C

by Dorin Nicolae Gheorghe, Dora Maria Popescu, Alex Salan, Mihail Virgil Boldeanu, Claudiu Marinel Ionele, Allma Pitru, Adina Turcu-Stiolica, Adrian Camen, Cristina Florescu, Ion Rogoveanu and Petra Surlin

J. Clin. Med. 2021, 10(22), 5275; https://doi.org/10.3390/jcm10225275 - 13 Nov 2021

Cited by 3 | Viewed by 2246

Abstract

Non-surgical periodontal therapy (NSPT) is the first essential step for the management of any periodontitis patient. This study aims to evaluate the impact of NSPT on pro-inflammatory mediators’ regulation and on clinical parameters in periodontitis patients who suffer from chronic hepatitis C. At [...] Read more.

Non-surgical periodontal therapy (NSPT) is the first essential step for the management of any periodontitis patient. This study aims to evaluate the impact of NSPT on pro-inflammatory mediators’ regulation and on clinical parameters in periodontitis patients who suffer from chronic hepatitis C. At baseline, selected patients were clinically evaluated for their periodontal status. A subsequent quantitative assessment of C-reactive protein and pentraxin-3 in samples of gingival fluid was performed by Enzyme-Linked Immunosorbent Assay (ELISA). Afterwards, NSPT was performed. Three months after NSPT, the clinical and ELISA assessments were repeated. The results show an improvement of the clinical parameters in periodontitis patients at the three-month recall. In chronic hepatitis C patients with periodontitis, the gingival fluid levels of pro-inflammatory markers reduced significantly. The targeted markers also expressed significant correlations with the clinical parameters used for the assessment of periodontitis’ severity. The results suggest that, while chronic hepatitis C patients exhibited a more negative periodontal status at baseline as compared to non-hepatitis ones, NSPT is effective in decreasing the local periodontal inflammatory reaction and in proving the periodontal status of this type of patients. Given the limitation of the study, periodontal screening and NSPT should be included in the integrated therapeutical approach of chronic hepatitis C patients, for its impact on the local inflammatory response. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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10 pages, 3469 KiB

Open AccessArticle

Humoral Response to Microbial Biomarkers in Rheumatoid Arthritis Patients

by Seyedesomaye Jasemi, Gian Luca Erre, Maria Luisa Cadoni, Marco Bo and Leonardo A. Sechi

J. Clin. Med. 2021, 10(21), 5153; https://doi.org/10.3390/jcm10215153 - 2 Nov 2021

Cited by 15 | Viewed by 2109

Abstract

Background/Objective: Chronic humoral immune response against multiple microbial antigens may play a crucial role in the etiopathogenesis of rheumatoid arthritis (RA). We aimed to assess the prevalence and magnitude of antibody response against various bacterial and viral immunogen peptides in the sera of [...] Read more.

Background/Objective: Chronic humoral immune response against multiple microbial antigens may play a crucial role in the etiopathogenesis of rheumatoid arthritis (RA). We aimed to assess the prevalence and magnitude of antibody response against various bacterial and viral immunogen peptides in the sera of RA patients compared with the general population. Methods: Polyclonal IgG antibodies (Abs) specific for peptides derived from Porphyromonas gingivalis (RgpA, Kpg), Aggregatibacter actinomycetemcomitans (LtxA1, LtxA2), Mycobacterium avium subsp. paratuberculosis (MAP4027), Epstein–Barr virus (EBNA1, EBVBOLF), and human endogenous retrovirus (HERV-W env-su) were detected by ELISA in serum samples from 148 consecutive RA patients and 148 sex and age-matched healthy controls (HCs). In addition, the presence of a relationship between the positivity and the titer of antibodies and RA descriptors was explored by bivariate correlation analysis. Results: RA patients exhibit a higher prevalence of humoral immune response against all tested peptides compared to HCs with a statically significant difference for MAP4027 (30.4% vs. 10.1%), BOLF (25.7% vs. 8.1%), RgpA (24.3% vs. 9.4%), HERV W-env (20.3% vs. 9.4%), and EBNA1 (18.9% vs. 9.4%) peptides. Fifty-three (35.8%) out of 148 RA serum and 93 (62.8%) out of 148 HCs were negative for all pathogen-derived peptides. There was a significant correlation between OD values obtained by ELISA test against all peptides (p < 0.0001). We also found an increased titer and prevalence of Abs against LtxA1 and LtxA2 in seropositive vs. seronegative RF (p = 0.019, p = 0.018). Conclusion: This study demonstrates a significantly increased humoral response against multiple pathogens in patients with RA and implies that they could be an important factor in the pathogenesis of the disease. Therefore, the role of each individual pathogen in RA needs to be further investigated. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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20 pages, 3139 KiB

Open AccessArticle

Association of Novel Advanced Glycation End-Product (AGE10) with Complications of Diabetes as Measured by Enzyme-Linked Immunosorbent Assay

by Agnieszka Bronowicka-Szydełko, Małgorzata Krzystek-Korpacka, Małgorzata Gacka, Jadwiga Pietkiewicz, Urszula Jakobsche-Policht and Andrzej Gamian

J. Clin. Med. 2021, 10(19), 4499; https://doi.org/10.3390/jcm10194499 - 29 Sep 2021

Cited by 11 | Viewed by 2571

Abstract

Advanced glycation end-products (AGEs) contribute to vascular complications and organ damage in diabetes. The unique AGE epitope (AGE10) has recently been identified in human serum using synthetic melibiose-derived AGE (MAGE). We aimed at developing ELISA for AGE10 quantification, determining whether AGE10 is present [...] Read more.

Advanced glycation end-products (AGEs) contribute to vascular complications and organ damage in diabetes. The unique AGE epitope (AGE10) has recently been identified in human serum using synthetic melibiose-derived AGE (MAGE). We aimed at developing ELISA for AGE10 quantification, determining whether AGE10 is present in diabetic patients (n = 82), and evaluating its association with diabetic complications. In a competitive ELISA developed, the reaction of synthetic MAGE with anti-MAGE was inhibited by physiological AGE10 present in serum. In this assay, new murine IgE anti-MAGE monoclonal antibodies, which do not recognize conventional AGEs, a synthetic MAGE used to coat the plate, and LMW-MAGE (low molecular mass MAGE) necessary to plot a standard curve were used. AGE10 was significantly higher in patients with microangiopathy, in whom it depended on treatment, being lower in patients treated with aspirin. AGE10 levels were positively correlated with estimated glomerular filtration rate (eGFR) and negatively with creatinine. As a marker of stage ≥3 chronic kidney disease or microangiopathy, AGE10 displayed moderate overall accuracy (respectively, 69% and 71%) and good sensitivity (82.6% and 83.3%) but poor specificity (58.1% and 57.8%). In conclusion, newly developed immunoassay allows for AGE10 quantification. AGE10 elevation is associated with microangiopathy while its decrease accompanies stage ≥3 chronic kidney disease. Full article

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13 pages, 509 KiB

Open AccessArticle

Does a Vegetarian Diet Affect the Levels of Myokine and Adipokine in Prepubertal Children?

by Jadwiga Ambroszkiewicz, Joanna Gajewska, Joanna Mazur, Witold Klemarczyk, Grażyna Rowicka, Mariusz Ołtarzewski, Małgorzata Strucińska and Magdalena Chełchowska

J. Clin. Med. 2021, 10(17), 3995; https://doi.org/10.3390/jcm10173995 - 3 Sep 2021

Cited by 7 | Viewed by 2481

Abstract

Myokines are cytokines secreted by muscle and exert autocrine, paracrine, or endocrine effects. Myokines mediate communication between muscle and other organs, including adipose tissue. The aim of the study was to assess serum myokines and their relationships with adipokines and anthropometric and nutritional [...] Read more.

Myokines are cytokines secreted by muscle and exert autocrine, paracrine, or endocrine effects. Myokines mediate communication between muscle and other organs, including adipose tissue. The aim of the study was to assess serum myokines and their relationships with adipokines and anthropometric and nutritional parameters in children following vegetarian and omnivorous diets. One hundred and five prepubertal children were examined. Among them there were 55 children on a vegetarian diet and 50 children on an omnivorous diet. Concentrations of myokines (myostatin, irisin) and adipokines (leptin, adiponectin, omentin, visfatin) in serum were determined by enzyme-linked immunosorbent assay (ELISA). We observed comparable median values of serum myokines and adipokines (except of leptin concentration) in both of the studied groups of children. We also found several correlations between myokine and adipokine levels and certain nutritional parameters. Serum myostatin was positively correlated with omentin levels in vegetarians and omnivores (p = 0.002). Serum irisin was positively associated with omentin (p = 0.045) levels in omnivores and inversely with visfatin concentration (p = 0.037) in vegetarians. Myostatin concentration was negatively correlated with the percentage of energy from protein (p = 0.014), calcium (p = 0.046), and vitamin A (p = 0.028) intakes in vegetarians and with dietary vitamin C (p = 0.041) and vitamin E (p = 0.021) intakes in omnivores. In multivariate regression analyses, positive correlations of serum myostatin with omentin levels were revealed in both study groups (β = 0.437, p < 0.001 for vegetarians; and β = 0.359, p = 0.001 for omnivores). Consuming a lacto-ovo-vegetarian diet did not influence serum levels of myokines (myostatin, irisin) and adipokines such as adiponectin, visfatin, and omentin in prepubertal children. However, leptin levels were significantly lower in vegetarians compared with omnivores. The observed significant positive correlations between myostatin and omentin concentrations might suggest tissue cross-talk between skeletal muscle and fat tissue. Further studies, carried out in a larger group of children following different dietary patterns, could be important to fully understand the relations between muscle, adipose tissues, and nutrition. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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37 pages, 8454 KiB

Open AccessArticle

The Paradoxical Effect Hypothesis of Abused Drugs in a Rat Model of Chronic Morphine Administration

by Yinghao Yu, Alan Bohan He, Michelle Liou, Chenyin Ou, Anna Kozłowska, Pingwen Chen and Andrew Chihwei Huang

J. Clin. Med. 2021, 10(15), 3197; https://doi.org/10.3390/jcm10153197 - 21 Jul 2021

Cited by 9 | Viewed by 3577

Abstract

A growing body of studies has recently shown that abused drugs could simultaneously induce the paradoxical effect in reward and aversion to influence drug addiction. However, whether morphine induces reward and aversion, and which neural substrates are involved in morphine’s reward and aversion [...] Read more.

A growing body of studies has recently shown that abused drugs could simultaneously induce the paradoxical effect in reward and aversion to influence drug addiction. However, whether morphine induces reward and aversion, and which neural substrates are involved in morphine’s reward and aversion remains unclear. The present study first examined which doses of morphine can simultaneously produce reward in conditioned place preference (CPP) and aversion in conditioned taste aversion (CTA) in rats. Furthermore, the aversive dose of morphine was determined. Moreover, using the aversive dose of 10 mg/kg morphine tested plasma corticosterone (CORT) levels and examined which neural substrates were involved in the aversive morphine-induced CTA on conditioning, extinction, and reinstatement. Further, we analyzed c-Fos and p-ERK expression to demonstrate the paradoxical effect—reward and aversion and nonhomeostasis or disturbance by morphine-induced CTA. The results showed that a dose of more than 20 mg/kg morphine simultaneously induced reward in CPP and aversion in CTA. A dose of 10 mg/kg morphine only induced the aversive CTA, and it produced higher plasma CORT levels in conditioning and reacquisition but not extinction. High plasma CORT secretions by 10 mg/kg morphine-induced CTA most likely resulted from stress-related aversion but were not a rewarding property of morphine. For assessments of c-Fos and p-ERK expression, the cingulate cortex 1 (Cg1), prelimbic cortex (PrL), infralimbic cortex (IL), basolateral amygdala (BLA), nucleus accumbens (NAc), and dentate gyrus (DG) were involved in the morphine-induced CTA, and resulted from the aversive effect of morphine on conditioning and reinstatement. The c-Fos data showed fewer neural substrates (e.g., PrL, IL, and LH) on extinction to be hyperactive. In the context of previous drug addiction data, the evidence suggests that morphine injections may induce hyperactivity in many neural substrates, which mediate reward and/or aversion due to disturbance and nonhomeostasis in the brain. The results support the paradoxical effect hypothesis of abused drugs. Insight from the findings could be used in the clinical treatment of drug addiction. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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17 pages, 8801 KiB

Open AccessArticle

Levels of Biological Markers of Nitric Oxide in Serum of Patients with Mandible Fractures

by Lukasz Wozniak, Wioletta Ratajczak-Wrona, Jan Borys, Bozena Antonowicz, Karolina Nowak, Piotr Bortnik and Ewa Jablonska

J. Clin. Med. 2021, 10(13), 2832; https://doi.org/10.3390/jcm10132832 - 26 Jun 2021

Cited by 4 | Viewed by 1930

Abstract

Background: Nitric oxide is a small gaseous molecule with significant bioactivity. It has been observed that NO may have a dual role dependent on its production and concentrations in the bone microenvironment. The objective of the study was to assess the concentration of [...] Read more.

Background: Nitric oxide is a small gaseous molecule with significant bioactivity. It has been observed that NO may have a dual role dependent on its production and concentrations in the bone microenvironment. The objective of the study was to assess the concentration of total nitric oxide malonyldialdehyde, nitrotyrosine, and asymmetric dimethylarginine in the serum of patients with mandibular fractures and to understand the relationship between these compounds, in order to expand the knowledge base of the role of nitric oxide and its activity indicators in the process of bone fracture healing. Material and Methods: The study included 20 patients with mandibular fractures who were undergoing inpatient and outpatient treatments and a control group of 15 healthy people. Results were analyzed with respect to the measurement time. Total nitric oxide concentration in the blood serum was determined according to the Griess reaction, while the concentration of malonyldialdehyde, nitrotyrosine, and asymmetric dimethylarginine was estimated using the immunoenzymatic method (i.e., enzyme-linked immunosorbent assay). Results: Before the procedure, as well as on the first day and 2 and 6 weeks after the procedure, higher concentrations of total nitric oxide and lower concentrations of malonyldialdehyde were observed in the blood serum of patients with mandibular fractures compared to the control group. No statistically significant differences were found in nitrotyrosine concentrations in the blood serum of patients throughout the measurement period. However, a significantly higher asymmetric dimethylarginine concentration was observed in the patient serum before the procedure and on the first day of operation as compared with the control group. Analysis of the results observed in patient serum with respect to the number of fractures within the mandible demonstrated the same trend of concentrations for the tested compounds for the entire study group. Conclusions: In summary, our results revealed that the intensity of local processes resulting from mandibular fractures is associated with the concentration of nitric oxide, confirming its significant role, as well as that of its indicators, in the process of bone fracture healing in this patient population. Full article

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12 pages, 2689 KiB

Open AccessArticle

Evaluation of a Broad Panel of SARS-CoV-2 Serological Tests for Diagnostic Use

by Maren Werner, Philip Pervan, Vivian Glück, Florian Zeman, Michael Koller, Ralph Burkhardt, Thomas Glück, Jürgen J. Wenzel, Barbara Schmidt, André Gessner and David Peterhoff

J. Clin. Med. 2021, 10(8), 1580; https://doi.org/10.3390/jcm10081580 - 8 Apr 2021

Cited by 10 | Viewed by 2683

Abstract

Serological testing is crucial in detection of previous infection and in monitoring convalescent and vaccine-induced immunity. During the Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) pandemic, numerous assay platforms have been developed and marketed for clinical use. Several studies recently compared clinical [...] Read more.

Serological testing is crucial in detection of previous infection and in monitoring convalescent and vaccine-induced immunity. During the Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) pandemic, numerous assay platforms have been developed and marketed for clinical use. Several studies recently compared clinical performance of a limited number of serological tests, but broad comparative evaluation is currently missing. Within this study, a panel of 161 sera from SARS-CoV-2 infected, seasonal CoV-infected and SARS-CoV-2 naïve subjects was enrolled to evaluate 16 ELISA/ECLIA-based and 16 LFA-based tests. Specificities of all ELISA/ECLIA-based assays were acceptable and generally in agreement with the providers’ specifications, but sensitivities were lower as specified. Results of the LFAs were less accurate as compared to the ELISAs, albeit with some exceptions. We found a sporadic unequal immune response for different antigens and thus recommend the use of a nucleocapsid protein (N)- and spike protein (S)-based test combination when maximal sensitivity is necessary. Finally, the quality of the immune response in terms of neutralization should be tested using S-based IgG tests. Full article

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Review

Jump to: Research, Other

13 pages, 1290 KiB

Open AccessReview

Enzyme-Linked Immunosorbent Assay: An Adaptable Methodology to Study SARS-CoV-2 Humoral and Cellular Immune Responses

by Amanda Izeli Portilho, Gabrielle Gimenes Lima and Elizabeth De Gaspari

J. Clin. Med. 2022, 11(6), 1503; https://doi.org/10.3390/jcm11061503 - 9 Mar 2022

Cited by 6 | Viewed by 4790

Abstract

The Enzyme-Linked Immunosorbent Assay is a versatile technique, which can be used for several applications. It has enormously contributed to the study of infectious diseases. This review highlights how this methodology supported the science conducted in COVID-19 pandemics, allowing scientists to better understand [...] Read more.

The Enzyme-Linked Immunosorbent Assay is a versatile technique, which can be used for several applications. It has enormously contributed to the study of infectious diseases. This review highlights how this methodology supported the science conducted in COVID-19 pandemics, allowing scientists to better understand the immune response against SARS-CoV-2. ELISA can be modified to assess the functionality of antibodies, as avidity and neutralization, respectively by the standardization of avidity-ELISA and surrogate-neutralization methods. Cellular immunity can also be studied using this assay. Products secreted by cells, like proteins and cytokines, can be studied by ELISA or its derivative Enzyme-linked immunospot (ELISpot) assay. ELISA and ELISA-based methods aided the area of immunology against infectious diseases and is still relevant, for example, as a promising approach to study the differences between natural and vaccine-induced immune responses against SARS-CoV-2. Full article

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13 pages, 1247 KiB

Open AccessReview

Modified ELISA for Ultrasensitive Diagnosis

by Naoko Tsurusawa, Jyunhao Chang, Mayuri Namba, Daiki Makioka, Sou Yamura, Kanako Iha, Yuta Kyosei, Satoshi Watabe, Teruki Yoshimura and Etsuro Ito

J. Clin. Med. 2021, 10(21), 5197; https://doi.org/10.3390/jcm10215197 - 7 Nov 2021

Cited by 25 | Viewed by 7060

Abstract

An enzyme-linked immunosorbent assay (ELISA) can be used for quantitative measurement of proteins, and improving the detection sensitivity to the ultrasensitive level would facilitate the diagnosis of various diseases. In the present review article, we first define the term ‘ultrasensitive’. We follow this [...] Read more.

An enzyme-linked immunosorbent assay (ELISA) can be used for quantitative measurement of proteins, and improving the detection sensitivity to the ultrasensitive level would facilitate the diagnosis of various diseases. In the present review article, we first define the term ‘ultrasensitive’. We follow this with a survey and discussion of the current literature regarding modified ELISA methods with ultrasensitive detection and their application for diagnosis. Finally, we introduce our own newly devised system for ultrasensitive ELISA combined with thionicotinamide adenine dinucleotide cycling and its application for the diagnosis of infectious diseases and lifestyle-related diseases. The aim of the present article is to expand the application of ultrasensitive ELISAs in the medical and biological fields. Full article

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Other

Jump to: Research, Review

8 pages, 5986 KiB

Open AccessBrief Report

Dynamic Features of Herd Immunity: Similarities in Age-Specific Anti-Measles Seroprevalence Data between Two Countries of Different Epidemiological History

by Katalin Böröcz, Senka Samardžić, Ines Drenjančević, Ákos Markovics, Tímea Berki and Péter Németh

J. Clin. Med. 2022, 11(4), 1145; https://doi.org/10.3390/jcm11041145 - 21 Feb 2022

Cited by 3 | Viewed by 2150

Abstract

(1) Background: Measles immunization gap(s) raise the concern of potential outbreaks. Both Croatia and Hungary are situated in the vicinity of measles-endemic countries. Potentially compromised immunization activities due to the COVID-19 surge is a ground for concern. Our aim was to compare age-stratified [...] Read more.

(1) Background: Measles immunization gap(s) raise the concern of potential outbreaks. Both Croatia and Hungary are situated in the vicinity of measles-endemic countries. Potentially compromised immunization activities due to the COVID-19 surge is a ground for concern. Our aim was to compare age-stratified seroprevalence results in the cross-border region. (2) Methods: Anti-MMR specific antibody levels (IgG) of 950 anonymous Croatian samples were compared with previous Hungarian results (n > 3500 samples), and former Croatian seroprevalence data (n = 1205). Seropositivity ratios were determined using our self-developed anti-MMR indirect ELISA (Euroimmun IgG ELISA kits were used as control). (3) Results: Measured seropositivity ratios of the Croatian samples were largely overlapping with our earlier published Hungarian data (the lowest seropositivity ratios were measured among individuals of 34–43 years of age with 78% of seropositivity) and are in accordance with earlier published data of Croatian researchers. (4) Conclusion: Although the epidemiological histories of the two countries are different, analogies in age-specific measles susceptibility have been discovered. We suggest that besides the potential coincidence in vaccination ineffectiveness, the inherent biological dynamics of vaccination-based humoral protection might have also contributed to the experienced similarities. Our findings may also serve as a lesson regarding the current anti-COVID-19 vaccination strategy. Full article

(This article belongs to the Special Issue Role of Enzyme-Linked Immunosorbent Assay (ELISA))

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