Startseite Evaluation of the performance of immunoblot and immunodot techniques used to identify autoantibodies in patients with autoimmune diseases
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Evaluation of the performance of immunoblot and immunodot techniques used to identify autoantibodies in patients with autoimmune diseases

  • Youssef EL Hassouni , Mohammed Bourhia EMAIL logo , Ahmed Bari , Riaz Ullah , Hafiz Majid Mahmood , Syed Saeed Ali , Samir Ibenmoussa und Admou Brahim
Veröffentlicht/Copyright: 2. März 2021
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Open Chemistry
Aus der Zeitschrift Open Chemistry Band 19 Heft 1

Abstract

Autoimmune diseases are pathological conditions in which the immune system mistakenly attacks its own tissues. This study evaluates the performance of two techniques, which are identifiers of autoantibody specifics: immunoblot and immunodot. This study was conducted in 300 patients of whom 62 were tested positive for antinuclear antibodies. The patients were initially screened for antinuclear antibodies using indirect immunofluorescence. Then, the identification of specific autoantibodies such as anti-extractable nuclear antigens (ENAs) was carried out using the immunoblot and immunodot techniques. The results showed that immunoblot and immunodot did not present a significant difference in their sensitivity against anti-SSA/52, SSB, CENP-B, PCNA, U1-snRNP, Jo-1, Pm-scl, and Mi-2 (p > 0.05). However, the two techniques showed a significant difference in their sensitivity toward autoantibodies anti-DNAn, anti-histone, anti-SmD1, and anti-ds-DNA (p < 0.05). The immunoblot data were in complete accordance with the immunodot data (100%) regarding the detection of autoantibodies such as anti SSA/52, SSB, CENP-B, PCNA, U1-snRP, Jo-1, Pm-scl, and Mi-2, 80% regarding SmD1, and 75% concerning ds-DNA. We should certainly pay closer attention to the efficiency of the techniques used in the diagnosis of autoimmune diseases.

Keywords: systemic autoimmune diseases; antinuclear antibodies; immunoblot; immunodot; indirect immunofluorescence

1 Introduction

Autoimmunity is a pathological state in which the body reacts, through a humoral (autoantibody) and cellular (self-reactive T cells) autoimmunity, against its own healthy cells and tissues [1]. Autoimmune diseases (ADs) affect around 5% of the world’s population and are the third leading cause of death in developed countries [2]. Autoimmune diseases are heterogeneous and are usually classified into two groups: specific autoimmune diseases and nonspecific autoimmune diseases. The first type habitually targets antigens located in a specific tissue or a cell, while the second type habitually targets antigens located anywhere in the body. Thus, specific and nonspecific autoimmune diseases are responsible for producing antinuclear antibodies (ANAs).

The antinuclear antibodies (ANAs) are autoantibodies that attack components of cells (anti-DNA, anti-nucleosome, anti-histone, and anti-RNA) triggering autoimmune disorders such as systemic lupus erythematosus [1]. Systemic lupus erythematosus is a nonspecific autoimmune disease that provokes damages in many organs and tissues of the body, such as the skin, heart, kidney, and nervous system. The most common symptoms of systemic lupus erythematosus are severe fatigue, joint pain, headache, hair loss, joint swelling, anemia, blood clotting, and appearance of a rash on the cheeks and nose, which is also known as a “butterfly rash” [3].

Autoantibodies, antibodies designated as SS-A or Ro and SS-B or La, were screened in patients with Sjögren’s syndrome (SS) and SLE and have been demonstrated to attack intracellular components that may be implicated with adjustment of RNA polymerase III function [4]. Autoantibodies in human sera are commonly identified by immunofluorescence microscopy (IFM). Tissue culture cells or tissue sections have been used as the antigen to identify autoantibodies under the immunofluorescence microscopy system. Even other assays such as enzyme-linked immunosorbent assay (ELISA) and Western blotting (WB) techniques have been used as alternatives to IFM in analyzing substrates; however, these assays will apparently not be a substitute for IFM [5].

The aim of this study is to develop immunological diagnosis methods for the laboratory of Immunology of CHU Morocco to be able to study the antinuclear antibodies released in the blood due to systemic autoimmune diseases. This study is particularly aimed to evaluate the performance of immunoblot and immunodot techniques used to identify specific autoantibodies, specifically the anti-extractable nuclear antigen (ENA).

2 Material and methods

This is a prospective study done for descriptive and analytical purposes. It was conducted at the Immunological laboratory of CHU Morocco, with 300 patients of whom 62 of them were tested positive for ANA. The identification of the ANA was carried out using immunoblot and immunodot assays.

The Human Ethics Review Committee of our University reviewed and approved this study. Furthermore, this study was performed according to the principles outlined in the Declaration of Helsinki, and institutional review boards and ethics committees at participating sites approved this protocol.

2.1 Sampling and preanalytical phase of testing

A total of 5 mL of blood was collected in tubes containing heparin from each patient. The preanalytical phase was conducted as follows: patient information was enrolled in the CHU informatics system, samples were labeled and centrifuged at 4,000 rpm for 10 min, and from each aliquot, 200 μL of plasma was recovered after centrifugation, and the aliquots were stored in a freezer at −20°C until usage.

2.2 Detection of autoantibodies using immunoblot technique (Blot-17)

At the first stage, ANA was detected by the indirect immunofluorescence assay in HEp-2 cells. This assay helps us to determine the appearance of ANA and orients it toward the antigenic targets. The next step was the identification of specific ANA with immunoblot.

2.2.1 Different components in the kit

ImmunoBlot Kit (AESKUBLOTS® ANA-17 Pro) was used to determine the type of detected ANA in HEp-2000 cells. The kit was purchased from GIGALAB Diagnostic and contained 24 numbered strips. Each strip consisted of a membrane on which different lines of highly purified autoantigens were placed.

2.2.2 Automated analyzer

The AESKUBLOTS® ANA-17 Protest was carried out using an Automated analyzer (HELMED-Automated Processor for ELISA, BLOT and IFA) (Figure 1).

Figure 1 Immunoblot image (Blot-17, HELMED).
Figure 1

Immunoblot image (Blot-17, HELMED).

2.2.3 Reading the results

The results were read using AESKU SCAN. This software was connected to a scanner that produces a sheet on which the strips are fixed. After reading the strips, the results were obtained in tables and peaks with positive and negative tests.

2.3 Detection of autoantibodies using immunodot technique (ANA-25 Screen IgG)

After identifying the autoantibody specificities using the immunoblot technique, the samples were then analyzed again using immunodot technique to compare the performance of the two techniques in identifying autoantibody specificities.

2.3.1 Automated analyzer

BlueDiver is an innovative instrument that allows the automation of different stages of the immunodot technique (Figure 2). The BlueDiver is characterized by reliability, speed, and low cost. Currently, it is used for the diagnosis of autoimmune diseases. The large combination of antigens offers a wide array that allows for the diagnosis of several autoimmune diseases such as conjunctivitis, gastritis, celiac disease, paraneoplastic syndromes, autoimmune hepatitis, primary biliary cholangitis, and vasculitis.

Figure 2 Immunodot image (BlueDiver).
Figure 2

Immunodot image (BlueDiver).

Dr-DOT is a multilingual software that allows for easy interpretation and semiquantitative or quantitative readings of BlueDot kit strips. This software requires only a computer and a BlueScan scanner. Dr-DOT software also contains a database that records the results (numerical values and scanned images).

2.4 Statistical analysis

Data were statistically analyzed using the t-test. A p value less than 0.05 was considered as statistically significant.

3 Results and discussion

3.1 Distribution of patients based on sex

The current study included patients with autoimmune disease including 75% females and 25% males.

This analytical and descriptive research study was conducted in 300 patients of whom 62 were positive for ANA according to the indirect immunofluorescence screening. The results showed a female predominance for positively testing for ANA with 75.81% (47 cases). These results were in agreement with the earlier data [6], where it was reported that the systemic autoimmune diseases exceedingly affected women of the reproductive age. The women’s predominance of antinuclear antibodies is explained by the influence of hormonal factors on the immune system. Indeed, estrogen stimulates the humoral immune response, while progesterone and androgens exert a suppressive effect on the humoral and cellular immune response. A high level of 17-beta estradiol was recorded in patients with rheumatoid arthritis and systemic lupus erythematosus [7].

3.2 Distribution of patients according to age

In this study, the age of the patients ranged from 1 to 71 years, with an average age of 41.92 years. Patients who aged between 31 and 46 years were the most affected with a frequency of 0.24 (24%) (Figure 3).

Figure 3 Distribution of patients according to age.
Figure 3

Distribution of patients according to age.

By analyzing the age categories of the studied patients, it was found that the age group between 31 and 46 years was the group most affected by ANA (24%). The study results were in agreement with those reported in early research data showing that systemic diseases frequently affects individuals with an average age of 42 years [8]. Systemic scleroderma (ScS) frequently affects individuals aged between 40 and 50 years [6].

3.3 Distribution of patients according to the type of disease

In the study population, four types of autoimmune diseases were detected. Lupus and Gougerot-Sjögren syndrome were the most frequent autoimmune diseases with frequencies of 40 and 23%, respectively (Figure 4).

Figure 4 Distribution of patients according to the type of disease.
Figure 4

Distribution of patients according to the type of disease.

In this study, lupus was found to be the most common disease with 40%, followed by Sjögren’s syndrome with 23%. The same results were reported in the previous literature, where it was shown that systemic lupus largely affects women during genital activity. An early study reported that patients older than 65 years were subject to ANA diagnosis. In this study, nobody was detected with the currently investigated diseases as reported in the previously published data [9]. According to the most recent data of pediatric rheumatology, juvenile idiopathic arthritis was 5–10 times less common than lupus in people aged 18 years or younger. In the case of the frequency of dermatomyositis in children, the annual incidence has been rarely established. It was estimated to be 0.37/1,00,000 in Finland and 6/1,00,000 in New York before the age of 15 years. According to the reported data, the incidence is around 10 to 20/1,00,000 and the prevalence ranged from 10 to 20/10,000 before the age of 18 years [3].

3.4 Immunological profile

All patients were tested positive for ANA according to indirect immunofluorescence screenings. Forty-eight percent (n = 29) of patients showed a speckled immunofluorescence (speckled aspect) and 27% (n = 16) showed a homogeneous immunofluorescence (homogeneous aspect) (Figure 5).

Figure 5 Distribution of antinuclear antibodies aspects.
Figure 5

Distribution of antinuclear antibodies aspects.

Regarding the immunological profile, an important diversity of immunofluorescence aspects was recorded with a predominance of speckled appearance with 48%, followed by the homogeneous aspect with 27%. Immunofluorescence aspects results point to the antibody specificity before the identification process [10].

Antinuclear antibodies are directed against nuclear constituents, anti-DNA, antinucleosome, antihistone, anti-SSA, and anti-SSB. Antinuclear antibodies were directed against polypeptides fixed in small RNA called YRNA, whether cytoplasmic (70%) or nuclear (30%). These ANAs are markers of lupus, systemic Sjögren syndrome, and undifferentiated connective tissue disease [11]. The reported findings in this study showed that anti-SSA was the most frequent in patients with lupus followed by anti-ds-DNA and anti-histone.

Gougerot-Sjögren syndrome was present in 23% of the studied population. Patients with anti-SSA were the most common (28%), followed by patients with anti-SSB (7%) and anti-histone. The current reports were in accordance with the previously published data showing that patients with anti-SSA antibodies represented 28% of the total affected cases. A total of 13.7% of patients were reported with anti-SSB antibodies. Conversely, anti-SSB and SSA antibodies were present in 94.5 and 54.5% cases, respectively [12].

A total of 76.9% of patients with Sjögren syndrome were positive for anti-SSA antibodies [13]. Anti-SSA and anti-SSB antibodies were identified in 7 and 9% of patients with Sjögren syndrome, respectively [14]. The results of this study reported that the anti-Scl-70 due to scleroderma positive tests were highly abundant with an index of 4.7. These findings were in agreement with the previously published results [15], in which it was reported that the frequency of anti-Scl-70 Ab was significantly abundant in patients younger than 50 years. Moreover, the results obtained in this study agree with those reported in the previous study, which showed that rheumatoid arthritis and autoimmune thyroiditis, which are severe autoimmune diseases, with serious antinuclear antibodies affect more than 5% of the population in developed countries [16,17].

3.5 Distribution of antinuclear antibodies according to systemic diseases

Anti-SSA antibodies were abundantly present in lupus patients, including nine cases with specific anti-SSA/60, six cases with anti-SSA/Ro 52 and anti-SSB, followed by five cases with anti-DNA and four cases with anti SmD1. In case of patients with Gougerot-Sjögren syndrome, two cases were registered with anti-SSA/60 and SSA/52 followed by two cases with anti-histones and one case with anti-SSB. In case of patients with systemic sclerosis, the only target in the present screening was one case with anti-Scl-70 (Figure 6).

Figure 6 Immunological results according to the presence of ANA in systemic diseases.
Figure 6

Immunological results according to the presence of ANA in systemic diseases.

3.6 Study of concordance between immunodot and the immunoblot results

No significant difference between immunodot and immunoblot sensitivity regarding eight antigenic targets including nine cases of SSA/Ro 52, five cases of SSB, two cases of CEN-PB, and a single case of Jo-1 and Mi-2 each (Table 1).

Table 1

Study of concordance and discordance between immunodot and immunoblot

Immunodot Immunoblot Concordance dot/blot (%) Discordance dot/blot (%)
Number of cases Percentage Number of cases Percentage
ds-DNA 8 13.00 6 9.60 75.00 25.00
Nucleosome 13 21 7 11 54 46
Histone 5 8 9 15 45 55
SmD1 5 8 4 6 80 20
PCNA 0 0 0 0 100 0
Rib-Po 2 3 1 2 50 50
SSA/60 13 21 11 18 85 15
SSA/Ro 52 9 15 9 15 100 0
SSB 5 8 5 8 100 0
CENP-B 2 3 2 3 100 0
Scl-70 2 3 1 2 50 50
U1-snRP 0 0 0 0 100 0
AMA-M2 3 5 1 2 33 67
Jo-1 1 2 1 2 100 0
Pm-scl 1 2 0 0 0 100
Mi-2 1 2 1 2 100 0
Ku 0 0 0 100 0

A significant difference between immunodot and immunoblot sensitivity toward the identification of autoantibodies was recorded for nine antigenic targets (p < 0.05): anti-ds-DNA with 25% (eight positive cases were detected by immunodot and six positive cases were detected by immunoblot), antinucleosomes with 46% (13 cases positive were detected by immunodot and seven were detected by immunoblot), anti-histones with 55% (five positive cases were detected by immunodot and nine positive cases were detected by immunoblot), anti-SmD1 with 20% (five positive cases were detected by immunodot and four cases were detected by immunoblot), anti-Rib-Po with 50% (two positive cases were detected by immunodot and one was detected by immunoblot), antis-Scl-70 with 50% (two positive cases were detected by immunodot and one was detected by immunoblot), AMA-M2 with 67% (three positive cases were detected by immunodot and one was detected by immunoblot), and anti-Pm-scl, with one case, was detected by immunodot.

This study aimed to compare Dot-25 and Blot-17. A significant difference was recorded between the performances of the two techniques. The results showed that the Blot-17 technique was more sensitive regarding certain investigated antinuclear antibodies including anti-histones. Conversely, Dot-25 technique was more sensitive versus other antinuclear antibodies (anti-SSA, anti-ds-DNA, and so on).

To the best of our knowledge, no previous study compared efficiency and performance of Dot-25 and Blot-17 techniques versus the same autoantibodies identification. Hence, this study seems to be the first study that highlighted the difference in the sensitivity of these techniques toward the same antinuclear antibodies.

The sensitivity and specificity of different tests used for the detection of autoantibodies can vary from one study to another according to several reasons: First, there is no international standardization of the methods used in the detection; thus, the normal values may vary according to the population study. Therefore, the diagnosis should not be established solely with a single method. The obtained results should always be interpreted taking into account the clinical examination conditions, the patient’s history, as well as the results obtained through other methods. No single technique can rule out the possibility of false-positive or false-negative results. Analysis with an indirect immunofluorescence test should be done to confirm the obtained findings.

Both Dot-25 and Blot-17 were equally sensitive against antinuclear antibodies (PCNA, SSA/52, SSB, CENP-B, Jo-1, Mi-2, and so on). There were no significant differences shown between their performances regarding the detection of the antinuclear antibodies. However, a significant difference was reported between the two techniques regarding other antinuclear antibodies including anti-nucleosome, anti-ds-DNA, anti-SSA/52, anti-SSB anti-histone, anti-SmD1, anti-SSA, and anti-ds-DNA (p > 0.05)).

4 Conclusion

This study was prospectively carried out to assess the sensitivity of immunoblot and immunodot versus the same antinuclear antibodies present in the serum of patients affected by systemic autoimmune diseases. Although both techniques, immunoblot and immunodot, are currently being used to identify the same variety of antinuclear antibodies, they do not have similar performance (sensitivity) versus the same antinuclear antibodies. Hence, it is very important to pay more attention to the performance of techniques used for the identification of antinuclear antibody.

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Acknowledgment

The authors extend their appreciation to the Deanship of Scientific Research at King Saud University for financial support through research group no. RG-1440-009.

  1. Competing interests: Riaz Ullah, who is the co-author of this article, is a current Editorial Board member of Open Chemistry. This fact did not affect the peer-review process. The authors declare no other conflict of interest.

  2. Funding: This research was finically supported by King Saud University through research group no. RG-1440-009.

  3. Author’s contribution: Y. E. H. and M. B.: writing original draft; A. B. and R. U.: formal analysis; H. M. M., S. S. A., S. I.: revision and editing; A. B.: methodology and supervision.

  4. Data availability statement: Data used to support the findings are incorporated in the manuscript.

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Received: 2020-04-30
Revised: 2020-12-22
Accepted: 2021-01-18
Published Online: 2021-03-02

© 2021 Youssef EL Hassouni et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  82. A comparative morphological study of titanium dioxide surface layer dental implants
  83. Changes in the antioxidative properties of honeys during their fermentation
  84. Erratum
  85. Erratum to “Corrosion study of copper in aqueous sulfuric acid solution in the presence of (2E,5E)-2,5-dibenzylidenecyclopentanone and (2E,5E)-bis[(4-dimethylamino)benzylidene]cyclopentanone: Experimental and theoretical study”
  86. Erratum to “Modified TDAE petroleum plasticiser”
  87. Corrigendum
  88. Corrigendum to “A nitric oxide-releasing prodrug promotes apoptosis in human renal carcinoma cells: Involvement of reactive oxygen species”
  89. Special Issue on 3rd IC3PE 2020
  90. Visible light-responsive photocatalyst of SnO2/rGO prepared using Pometia pinnata leaf extract
  91. Antihyperglycemic activity of Centella asiatica (L.) Urb. leaf ethanol extract SNEDDS in zebrafish (Danio rerio)
  92. Selection of oil extraction process from Chlorella species of microalgae by using multi-criteria decision analysis technique for biodiesel production
  93. Special Issue on the 14th Joint Conference of Chemistry (14JCC)
  94. Synthesis and in vitro cytotoxicity evaluation of isatin-pyrrole derivatives against HepG2 cell line
  95. CO2 gas separation using mixed matrix membranes based on polyethersulfone/MIL-100(Al)
  96. Effect of synthesis and activation methods on the character of CoMo/ultrastable Y-zeolite catalysts
  97. Special Issue on Electrochemical Amplified Sensors
  98. Enhancement of graphene oxide through β-cyclodextrin composite to sensitive analysis of an antidepressant: Sulpiride
  99. Investigation of the spectroelectrochemical behavior of quercetin isolated from Zanthoxylum bungeanum
  100. An electrochemical sensor for high sensitive determination of lysozyme based on the aptamer competition approach
  101. An improved non-enzymatic electrochemical sensor amplified with CuO nanostructures for sensitive determination of uric acid
  102. Special Issue on Applied Biochemistry and Biotechnology 2020
  103. Fast discrimination of avocado oil for different extracted methods using headspace-gas chromatography-ion mobility spectroscopy with PCA based on volatile organic compounds
  104. Effect of alkali bases on the synthesis of ZnO quantum dots
  105. Quality evaluation of Cabernet Sauvignon wines in different vintages by 1H nuclear magnetic resonance-based metabolomics
  106. Special Issue on the Joint Science Congress of Materials and Polymers (ISCMP 2019)
  107. Diatomaceous Earth: Characterization, thermal modification, and application
  108. Electrochemical determination of atenolol and propranolol using a carbon paste sensor modified with natural ilmenite
  109. Special Issue on the Conference of Energy, Fuels, Environment 2020
  110. Assessment of the mercury contamination of landfilled and recovered foundry waste – a case study
  111. Primary energy consumption in selected EU Countries compared to global trends
  112. Modified TDAE petroleum plasticiser
  113. Use of glycerol waste in lactic acid bacteria metabolism for the production of lactic acid: State of the art in Poland
  114. Topical Issue on Applications of Mathematics in Chemistry
  115. Theoretical study of energy, inertia and nullity of phenylene and anthracene
  116. Banhatti, revan and hyper-indices of silicon carbide Si2C3-III[n,m]
  117. Topical Issue on Agriculture
  118. Occurrence of mycotoxins in selected agricultural and commercial products available in eastern Poland
  119. Special Issue on Ethnobotanical, Phytochemical and Biological Investigation of Medicinal Plants
  120. Acute and repeated dose 60-day oral toxicity assessment of chemically characterized Berberis hispanica Boiss. and Reut in Wistar rats
  121. Phytochemical profile, in vitro antioxidant, and anti-protein denaturation activities of Curcuma longa L. rhizome and leaves
  122. Antiplasmodial potential of Eucalyptus obliqua leaf methanolic extract against Plasmodium vivax: An in vitro study
  123. Prunus padus L. bark as a functional promoting component in functional herbal infusions – cyclooxygenase-2 inhibitory, antioxidant, and antimicrobial effects
  124. Molecular and docking studies of tetramethoxy hydroxyflavone compound from Artemisia absinthium against carcinogens found in cigarette smoke
  125. Special Issue on the Joint Science Congress of Materials and Polymers (ISCMP 2020)
  126. Preparation of cypress (Cupressus sempervirens L.) essential oil loaded poly(lactic acid) nanofibers
  127. Influence of mica mineral on flame retardancy and mechanical properties of intumescent flame retardant polypropylene composites
  128. Production and characterization of thermoplastic elastomer foams based on the styrene–ethylene–butylene–styrene (SEBS) rubber and thermoplastic material
  129. Special Issue on Applied Chemistry in Agriculture and Food Science
  130. Impact of essential oils on the development of pathogens of the Fusarium genus and germination parameters of selected crops
  131. Yield, volume, quality, and reduction of biotic stress influenced by titanium application in oilseed rape, winter wheat, and maize cultivations
  132. Influence of potato variety on polyphenol profile composition and glycoalcaloid contents of potato juice
  133. Carryover effect of direct-fed microbial supplementation and early weaning on the growth performance and carcass characteristics of growing Najdi lambs
  134. Special Issue on Applied Biochemistry and Biotechnology (ABB 2021)
  135. The electrochemical redox mechanism and antioxidant activity of polyphenolic compounds based on inlaid multi-walled carbon nanotubes-modified graphite electrode
  136. Study of an adsorption method for trace mercury based on Bacillus subtilis
  137. Special Issue on The 1st Malaysia International Conference on Nanotechnology & Catalysis (MICNC2021)
  138. Mitigating membrane biofouling in biofuel cell system – A review
  139. Mechanical properties of polymeric biomaterials: Modified ePTFE using gamma irradiation
https://doi.org/10.1515/chem-2020-0101
Schlagwörter für diesen Artikel
systemic autoimmune diseases; antinuclear antibodies; immunoblot; immunodot; indirect immunofluorescence
Creative Commons
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Artikel in diesem Heft

  1. Regular Articles
  2. Qualitative and semi-quantitative assessment of anthocyanins in Tibetan hulless barley from different geographical locations by UPLC-QTOF-MS and their antioxidant capacities
  3. Effect of sodium chloride on the expression of genes involved in the salt tolerance of Bacillus sp. strain “SX4” isolated from salinized greenhouse soil
  4. GC-MS analysis of mango stem bark extracts (Mangifera indica L.), Haden variety. Possible contribution of volatile compounds to its health effects
  5. Influence of nanoscale-modified apatite-type calcium phosphates on the biofilm formation by pathogenic microorganisms
  6. Removal of paracetamol from aqueous solution by containment composites
  7. Investigating a human pesticide intoxication incident: The importance of robust analytical approaches
  8. Induction of apoptosis and cell cycle arrest by chloroform fraction of Juniperus phoenicea and chemical constituents analysis
  9. Recovery of γ-Fe2O3 from copper ore tailings by magnetization roasting and magnetic separation
  10. Effects of different extraction methods on antioxidant properties of blueberry anthocyanins
  11. Modeling the removal of methylene blue dye using a graphene oxide/TiO2/SiO2 nanocomposite under sunlight irradiation by intelligent system
  12. Antimicrobial and antioxidant activities of Cinnamomum cassia essential oil and its application in food preservation
  13. Full spectrum and genetic algorithm-selected spectrum-based chemometric methods for simultaneous determination of azilsartan medoxomil, chlorthalidone, and azilsartan: Development, validation, and application on commercial dosage form
  14. Evaluation of the performance of immunoblot and immunodot techniques used to identify autoantibodies in patients with autoimmune diseases
  15. Computational studies by molecular docking of some antiviral drugs with COVID-19 receptors are an approach to medication for COVID-19
  16. Synthesis of amides and esters containing furan rings under microwave-assisted conditions
  17. Simultaneous removal efficiency of H2S and CO2 by high-gravity rotating packed bed: Experiments and simulation
  18. Design, synthesis, and biological activities of novel thiophene, pyrimidine, pyrazole, pyridine, coumarin and isoxazole: Dydrogesterone derivatives as antitumor agents
  19. Content and composition analysis of polysaccharides from Blaps rynchopetera and its macrophage phagocytic activity
  20. A new series of 2,4-thiazolidinediones endowed with potent aldose reductase inhibitory activity
  21. Assessing encapsulation of curcumin in cocoliposome: In vitro study
  22. Rare norisodinosterol derivatives from Xenia umbellata: Isolation and anti-proliferative activity
  23. Comparative study of antioxidant and anticancer activities and HPTLC quantification of rutin in white radish (Raphanus sativus L.) leaves and root extracts grown in Saudi Arabia
  24. Comparison of adsorption properties of commercial silica and rice husk ash (RHA) silica: A study by NIR spectroscopy
  25. Sodium borohydride (NaBH4) as a high-capacity material for next-generation sodium-ion capacitors
  26. Aroma components of tobacco powder from different producing areas based on gas chromatography ion mobility spectrometry
  27. The effects of salinity on changes in characteristics of soils collected in a saline region of the Mekong Delta, Vietnam
  28. Synthesis, properties, and activity of MoVTeNbO catalysts modified by zirconia-pillared clays in oxidative dehydrogenation of ethane
  29. Synthesis and crystal structure of N,N′-bis(4-chlorophenyl)thiourea N,N-dimethylformamide
  30. Quantitative analysis of volatile compounds of four Chinese traditional liquors by SPME-GC-MS and determination of total phenolic contents and antioxidant activities
  31. A novel separation method of the valuable components for activated clay production wastewater
  32. On ve-degree- and ev-degree-based topological properties of crystallographic structure of cuprite Cu2O
  33. Antihyperglycemic effect and phytochemical investigation of Rubia cordifolia (Indian Madder) leaves extract
  34. Microsphere molecularly imprinted solid-phase extraction for diazepam analysis using itaconic acid as a monomer in propanol
  35. A nitric oxide-releasing prodrug promotes apoptosis in human renal carcinoma cells: Involvement of reactive oxygen species
  36. Machine vision-based driving and feedback scheme for digital microfluidics system
  37. Study on the application of a steam-foam drive profile modification technology for heavy oil reservoir development
  38. Ni–Ru-containing mixed oxide-based composites as precursors for ethanol steam reforming catalysts: Effect of the synthesis methods on the structural and catalytic properties
  39. Preparation of composite soybean straw-based materials by LDHs modifying as a solid sorbent for removal of Pb(ii) from water samples
  40. Synthesis and spectral characterizations of vanadyl(ii) and chromium(iii) mixed ligand complexes containing metformin drug and glycine amino acid
  41. In vitro evaluation of lactic acid bacteria with probiotic activity isolated from local pickled leaf mustard from Wuwei in Anhui as substitutes for chemical synthetic additives
  42. Utilization and simulation of innovative new binuclear Co(ii), Ni(ii), Cu(ii), and Zn(ii) diimine Schiff base complexes in sterilization and coronavirus resistance (Covid-19)
  43. Phosphorylation of Pit-1 by cyclin-dependent kinase 5 at serine 126 is associated with cell proliferation and poor prognosis in prolactinomas
  44. Molecularly imprinted membrane for transport of urea, creatinine, and vitamin B12 as a hemodialysis candidate membrane
  45. Optimization of Murrayafoline A ethanol extraction process from the roots of Glycosmis stenocarpa, and evaluation of its Tumorigenesis inhibition activity on Hep-G2 cells
  46. Highly sensitive determination of α-lipoic acid in pharmaceuticals on a boron-doped diamond electrode
  47. Synthesis, chemo-informatics, and anticancer evaluation of fluorophenyl-isoxazole derivatives
  48. In vitro and in vivo investigation of polypharmacology of propolis extract as anticancer, antibacterial, anti-inflammatory, and chemical properties
  49. Topological indices of bipolar fuzzy incidence graph
  50. Preparation of Fe3O4@SiO2–ZnO catalyst and its catalytic synthesis of rosin glycol ester
  51. Construction of a new luminescent Cd(ii) compound for the detection of Fe3+ and treatment of Hepatitis B
  52. Investigation of bovine serum albumin aggregation upon exposure to silver(i) and copper(ii) metal ions using Zetasizer
  53. Discoloration of methylene blue at neutral pH by heterogeneous photo-Fenton-like reactions using crystalline and amorphous iron oxides
  54. Optimized extraction of polyphenols from leaves of Rosemary (Rosmarinus officinalis L.) grown in Lam Dong province, Vietnam, and evaluation of their antioxidant capacity
  55. Synthesis of novel thiourea-/urea-benzimidazole derivatives as anticancer agents
  56. Potency and selectivity indices of Myristica fragrans Houtt. mace chloroform extract against non-clinical and clinical human pathogens
  57. Simple modifications of nicotinic, isonicotinic, and 2,6-dichloroisonicotinic acids toward new weapons against plant diseases
  58. Synthesis, optical and structural characterisation of ZnS nanoparticles derived from Zn(ii) dithiocarbamate complexes
  59. Presence of short and cyclic peptides in Acacia and Ziziphus honeys may potentiate their medicinal values
  60. The role of vitamin D deficiency and elevated inflammatory biomarkers as risk factors for the progression of diabetic nephropathy in patients with type 2 diabetes mellitus
  61. Quantitative structure–activity relationship study on prolonged anticonvulsant activity of terpene derivatives in pentylenetetrazole test
  62. GADD45B induced the enhancing of cell viability and proliferation in radiotherapy and increased the radioresistance of HONE1 cells
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  64. Dynamics of λ-cyhalothrin disappearance and expression of selected P450 genes in bees depending on the ambient temperature
  65. Identification of synthetic cannabinoid methyl 2-{[1-(cyclohexylmethyl)-1H-indol-3-yl] formamido}-3-methylbutanoate using modern mass spectrometry and nuclear magnetic resonance techniques
  66. Study on the speciation of arsenic in the genuine medicinal material honeysuckle
  67. Two Cu(ii)-based coordination polymers: Crystal structures and treatment activity on periodontitis
  68. Conversion of furfuryl alcohol to ethyl levulinate in the presence of mesoporous aluminosilicate catalyst
  69. Review Articles
  70. Hsien Wu and his major contributions to the chemical era of immunology
  71. Overview of the major classes of new psychoactive substances, psychoactive effects, analytical determination and conformational analysis of selected illegal drugs
  72. An overview of persistent organic pollutants along the coastal environment of Kuwait
  73. Mechanism underlying sevoflurane-induced protection in cerebral ischemia–reperfusion injury
  74. COVID-19 and SARS-CoV-2: Everything we know so far – A comprehensive review
  75. Challenge of diabetes mellitus and researchers’ contributions to its control
  76. Advances in the design and application of transition metal oxide-based supercapacitors
  77. Color and composition of beauty products formulated with lemongrass essential oil: Cosmetics formulation with lemongrass essential oil
  78. The structural chemistry of zinc(ii) and nickel(ii) dithiocarbamate complexes
  79. Bioprospecting for antituberculosis natural products – A review
  80. Recent progress in direct urea fuel cell
  81. Rapid Communications
  82. A comparative morphological study of titanium dioxide surface layer dental implants
  83. Changes in the antioxidative properties of honeys during their fermentation
  84. Erratum
  85. Erratum to “Corrosion study of copper in aqueous sulfuric acid solution in the presence of (2E,5E)-2,5-dibenzylidenecyclopentanone and (2E,5E)-bis[(4-dimethylamino)benzylidene]cyclopentanone: Experimental and theoretical study”
  86. Erratum to “Modified TDAE petroleum plasticiser”
  87. Corrigendum
  88. Corrigendum to “A nitric oxide-releasing prodrug promotes apoptosis in human renal carcinoma cells: Involvement of reactive oxygen species”
  89. Special Issue on 3rd IC3PE 2020
  90. Visible light-responsive photocatalyst of SnO2/rGO prepared using Pometia pinnata leaf extract
  91. Antihyperglycemic activity of Centella asiatica (L.) Urb. leaf ethanol extract SNEDDS in zebrafish (Danio rerio)
  92. Selection of oil extraction process from Chlorella species of microalgae by using multi-criteria decision analysis technique for biodiesel production
  93. Special Issue on the 14th Joint Conference of Chemistry (14JCC)
  94. Synthesis and in vitro cytotoxicity evaluation of isatin-pyrrole derivatives against HepG2 cell line
  95. CO2 gas separation using mixed matrix membranes based on polyethersulfone/MIL-100(Al)
  96. Effect of synthesis and activation methods on the character of CoMo/ultrastable Y-zeolite catalysts
  97. Special Issue on Electrochemical Amplified Sensors
  98. Enhancement of graphene oxide through β-cyclodextrin composite to sensitive analysis of an antidepressant: Sulpiride
  99. Investigation of the spectroelectrochemical behavior of quercetin isolated from Zanthoxylum bungeanum
  100. An electrochemical sensor for high sensitive determination of lysozyme based on the aptamer competition approach
  101. An improved non-enzymatic electrochemical sensor amplified with CuO nanostructures for sensitive determination of uric acid
  102. Special Issue on Applied Biochemistry and Biotechnology 2020
  103. Fast discrimination of avocado oil for different extracted methods using headspace-gas chromatography-ion mobility spectroscopy with PCA based on volatile organic compounds
  104. Effect of alkali bases on the synthesis of ZnO quantum dots
  105. Quality evaluation of Cabernet Sauvignon wines in different vintages by 1H nuclear magnetic resonance-based metabolomics
  106. Special Issue on the Joint Science Congress of Materials and Polymers (ISCMP 2019)
  107. Diatomaceous Earth: Characterization, thermal modification, and application
  108. Electrochemical determination of atenolol and propranolol using a carbon paste sensor modified with natural ilmenite
  109. Special Issue on the Conference of Energy, Fuels, Environment 2020
  110. Assessment of the mercury contamination of landfilled and recovered foundry waste – a case study
  111. Primary energy consumption in selected EU Countries compared to global trends
  112. Modified TDAE petroleum plasticiser
  113. Use of glycerol waste in lactic acid bacteria metabolism for the production of lactic acid: State of the art in Poland
  114. Topical Issue on Applications of Mathematics in Chemistry
  115. Theoretical study of energy, inertia and nullity of phenylene and anthracene
  116. Banhatti, revan and hyper-indices of silicon carbide Si2C3-III[n,m]
  117. Topical Issue on Agriculture
  118. Occurrence of mycotoxins in selected agricultural and commercial products available in eastern Poland
  119. Special Issue on Ethnobotanical, Phytochemical and Biological Investigation of Medicinal Plants
  120. Acute and repeated dose 60-day oral toxicity assessment of chemically characterized Berberis hispanica Boiss. and Reut in Wistar rats
  121. Phytochemical profile, in vitro antioxidant, and anti-protein denaturation activities of Curcuma longa L. rhizome and leaves
  122. Antiplasmodial potential of Eucalyptus obliqua leaf methanolic extract against Plasmodium vivax: An in vitro study
  123. Prunus padus L. bark as a functional promoting component in functional herbal infusions – cyclooxygenase-2 inhibitory, antioxidant, and antimicrobial effects
  124. Molecular and docking studies of tetramethoxy hydroxyflavone compound from Artemisia absinthium against carcinogens found in cigarette smoke
  125. Special Issue on the Joint Science Congress of Materials and Polymers (ISCMP 2020)
  126. Preparation of cypress (Cupressus sempervirens L.) essential oil loaded poly(lactic acid) nanofibers
  127. Influence of mica mineral on flame retardancy and mechanical properties of intumescent flame retardant polypropylene composites
  128. Production and characterization of thermoplastic elastomer foams based on the styrene–ethylene–butylene–styrene (SEBS) rubber and thermoplastic material
  129. Special Issue on Applied Chemistry in Agriculture and Food Science
  130. Impact of essential oils on the development of pathogens of the Fusarium genus and germination parameters of selected crops
  131. Yield, volume, quality, and reduction of biotic stress influenced by titanium application in oilseed rape, winter wheat, and maize cultivations
  132. Influence of potato variety on polyphenol profile composition and glycoalcaloid contents of potato juice
  133. Carryover effect of direct-fed microbial supplementation and early weaning on the growth performance and carcass characteristics of growing Najdi lambs
  134. Special Issue on Applied Biochemistry and Biotechnology (ABB 2021)
  135. The electrochemical redox mechanism and antioxidant activity of polyphenolic compounds based on inlaid multi-walled carbon nanotubes-modified graphite electrode
  136. Study of an adsorption method for trace mercury based on Bacillus subtilis
  137. Special Issue on The 1st Malaysia International Conference on Nanotechnology & Catalysis (MICNC2021)
  138. Mitigating membrane biofouling in biofuel cell system – A review
  139. Mechanical properties of polymeric biomaterials: Modified ePTFE using gamma irradiation
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