Comparison of Serum Levels of SIL-2R, IL-6, IL-10, TNF-α, CRP, ESR and Fibrinogen in Patients with Active and Inactive Behçet’s Disease

Research Article

Comparison of Serum Levels of SIL-2R, IL-6, IL-10, TNF-α, CRP, ESR and Fibrinogen in Patients with Active and Inactive Behçet’s Disease

  • Ali Osman AVCI *

*Corresponding Author: Ali Osman AVCI, Gülhane Askerî Tıp Akademisi, Turkey.

Citation: Ali Osman AVCI, (2023), Comparison of serum levels of SIL-2R, IL-6, IL-10, TNF-α, CRP, ESR and Fibrinogen in Patients with Active and Inactive Behçet’s Disease, International Journal of Clinical Case Reports and Reviews, 13(5); DOI: 10.31579/2690-4861/312

Copyright: © 2023, Ali Osman AVCI. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Received: 09 May 2023 | Accepted: 22 May 2023 | Published: 31 May 2023

Keywords: Behçet’s Disease; SIL-2R; IL-6; IL-10; TNF-α

Abstract

Behçet's disease (BD) is a chronic inflammatory illness that affects the entire body and is characterized by recurring episodes of oral aphthae, ocular and cutaneous lesions, and scrotal or vaginal ulcerations. The involvement of other organs and systems increases mortality in addition to the significant morbidity.

This study involved a total of 40 participants, 20 of whom were healthy controls and 20 of whom were patients (of the 20 Behçet's patients, 12 (60%) were in the active phase and 8 (40%) were in the inactive phase). There was no therapy being given to any of the 20 patients that would have affected their cytokine levels.

Only young men made up the patient and control groups because both early onset and male sex are signs of poor prognosis.

The ELISA method was used to measure the levels of serum cytokines. The statistical analysis of the derived numerical values employed the Mann-Whitney U Test.

We found a significant correlation between serum cytokine levels and classical acute phase markers in active Behçet’s patients. ESR (P < 0,001), CRP (P < 0,001), fibrinogen (P < 0,001), IL-10 (P < 0,001), IL-6 (P < 0,001), SIL-2R (P < 0,001) and TNFα (P < 0,001).

There was no statistically significant difference in serum levels of classical acute phase markers ESR (P = 0,746), CRP (P = 0,476) and fibrinogen (P = 0,940) when inactive Behçet’s patients and healthy controls were compared. However, serum levels of IL-10 (P < 0,001), IL-6 (P = 0,001), SIL-2R (P < 0,001) and TNFα (P = 0,001) were statistically different between inactive Behcet's patients and the control group.

Our research shows that even in the inactive phase, serum cytokine levels of Behçet’s patients are much higher than the healthy control group. However, the levels of ESR, CRP and fibrinogen, which are classical acute phase markers, were found at normal levels in Behçet’s patients in the inactive phase. These findings show that measurement of serum interleukin levels will enable us to take preventive measures for morbidity and mortality follow-up of Behçet's patients.

Introduction

Behçet's disease (BD) is a systemic, long-term inflammatory condition marked by recurring episodes of oral aphthae, cutaneous, ophthalmic, and scrotal or vaginal lesions. Vascular, articular, gastrointestinal, cardiac, pulmonary, renal, and neurologic dysfunction are all linked to Behçet's Disease. Despite the fact that the discoveries date back to Hippocrates, Turkish Professor Dr. Hulusi Behçet was the first to describe it as a distinct illness in 1937 [1, 2].

The "Silk Road," the Middle East, the Mediterranean coast, and Central Asia all have higher rates of BD. The disease's greatest documented prevalence, which ranges from 110 to 420 cases per 100,000 individuals, was originally noted in Turkey. BD is present practically everywhere in the world, despite the fact that regional prevalence varies greatly. The disease, which affects both sexes equally, is the most It often occurs between the ages of 20–40. Male sex and disease early onset as causes of poor prognosis accepted [2, 3].

The exact cause of BD is not known yet. The theory that BD is an aberrant immune response that is brought on by bacterial, viral, and other environmental antigens or autoantigens like heat shock proteins (HSPs) is the one that is now most widely supported [4–7].

Only clinical signs and symptoms can be used to make the diagnosis of BD because there isn't a universally recognized diagnostic laboratory test for the condition. The most widely used and widely acknowledged diagnostic criteria for Behçet's disease are the International Working Group for the criteria for BD (the presence of either typical eye lesions, typical cutaneous lesions, or a positive skin pathergy test in addition to oral ulcers and recurrent genital ulcers). Increased serum levels of biochemical factors including CRP and cytokines have been suggested as indicators of disease activity [8–12].

In order to better understand the etiopathogenesis of BD, numerous cytokines have been studied. Regarding the blood cytokine levels of BP in the active and inactive stages, these investigations found remarkably disparate findings.

Patients with BD have higher levels of proinflammatory cytokines, particularly when the disease is active. The enhanced inflammatory response is thought to be caused by this elevation, particularly those implicated in Th1-mediated inflammation, along with genetic susceptibility. The SlL-2 receptor is thought to contribute to local inflammation. (13,14). TNFα enhances all types of humoral and cellular immune response and acts synergistically with IL-6 [15, 16].

Since IL-10 inhibits the production of various cytokines, IL-10 was designated as “Cytokine Synthesis Inhibitory Factor” (CSIF) when it was first identified. However, IL-10 reduces inflammation by reducing nitric oxide synthase induction and toxic oxygen radicals and monocyte prostaglandin H synthase-2 (COX-2) in macrophages [17–20].

It has been reported that circulating levels of SIL-2R, TNFα, and IL-6 are increased in Behçet’s patients [21]. It has been suggested that the increase in serum SIL-2R, TNFα and IL-6 levels is more pronounced in active-stage Behçet's patients.

It was also found that the increase in serum SIL-2R and IL-6 levels was more pronounced in BP with complications; It has been reported that IL-6 levels increase in the cerebrospinal fluid of patients with neuro-Behçet's disease. There are also studies reporting that serum SIL-2R, IL-6, IL-4, IL-10 and TNFα levels in Behçet’s patients do not increase in both active and inactive periods. Serum cytokine levels and conventional acute phase reactants have also been compared in several research [22–27].

Material and Methods

This study was conducted to compare serum SIL-2R, IL-6, IL-10, TNF-α levels with serum CRP, ESR and fibrinogen levels in active and inactive young male Behçet's patients.

A total of 40 persons participated in this study, 20 of whom were Behçet’s Patients and 20 of whom were healthy controls. Only young men made up the patient and control groups because both early onset and male sex are signs of poor prognosis.

The patient and control groups were selected from among the young men who came for enlistment examinations. A control group was formed from those who did not have any health problems or complaints and had no pathological findings according to the results of physical examination and blood tests. Those who had a history of Behçet's disease and still had active findings and did not use drugs constituted the active patient group, and those without active findings constituted the inactive patient group. Those currently using any medication were not included in any group.

The patients with Behçet ranged in age from 20 to 30, with a mean age of 22.55 years. Their diagnoses were made in accordance with the clinical standards established by the "International Study Group". Twelve (12%) of the 20 Behçet’s patients were included in the trial during the inactive phase, while eighty (40%) were in the active phase. Patients with at least two of the oral ulcers, genital ulcers, eye lesions and active arthritis were considered active patients.

Healthy men in the same age range made up the control group. The control group's age range was 20 to 29, and the average age was 22.45 years.

Blood samples were taken from the patient and control groups in the morning, fasting, into sterile tubes for ESR, CRP, fibrinogen, cytokine levels (IL-6, IL-10, SIL-2R and TNFα). Blood samples taken for cytokine levels were centrifuged sterilely at 3000 rpm for 10 minutes and their serums were separated. The serums were placed in sterile tubes with patient names on them and stored in deep-freeze at -40°C.

Determination of Soluble Interleukin-2 Receptor: Serum SIL-2R level was measured using commercially available ELISA kit (CYTELISA; Cytimmunesciences, Paris, France).

  1. The samples to be studied and the kit were brought to room temperature.
  2. 100 µl of standard, patient sample and control materials were placed in each well.
  3. 25 µl of Anti-Human SIL-2R monoclonal antibody was added to them.
  4. In order to prevent evaporation, the plates were covered with Acetate Plate Sealer and incubated for 3 hours at room temperature.
  5. The solutions in the wells were aspirated and emptied.
  6. The wells were filled with 250 ml of washing solution, waited for 15–30 seconds and aspirated. This process was repeated 3–4 times. Plates were dried by inversion.
  7. 50 µl of Anti-Rabbit conjugated Alkaline phosphatase was added to all wells.
  8. The steps 5 and 6 were repeated.
  9. 200 µl of the prepared coloring solution was added.
  10. Plates were closed and incubated for 15 minutes at room temperature.
  11. Plates were read at a wavelength of 492 nanometers.

Obtained results were evaluated by comparison with standard calibrators. The optical densities of the calibrators were analyzed with the graph obtained by performing linear-log regression analysis. The minimum SIL-2R level that the test can measure is 5 pg/ml. was in the form.

Tumor Necrosis Factor-α determination: Serum TNFα level was measured using commercially available ELISA kit (CYTELISA; Cytimmune sciences, Paris, France).

  1. The samples to be studied and the kit were brought to room temperature.
  2. 100 µl of standard, patient sample and control materials were placed in each well.
  3. 25 µl of Anti-Human TNFα monoclonal antibody was added to them.
  4. In order to prevent evaporation, the plates were covered with Acetate Plate Sealer and incubated for 3 hours at room temperature.
  5. The solutions in the wells were aspirated and emptied.
  6. The wells were filled with 250 ml of washing solution, waited for 15–30 seconds and aspirated. This process was repeated 3–4 times. Plates were dried by inversion.
  7. 50 µl of Anti-Rabbit Conjugated Alkaline Phosphatase was Added to all wells.
  8. The steps 5 and 6 were repeated.
  9. 200 µl of the prepared coloring solution was added.
  10. Plates were closed and incubated for 15 minutes at room temperature.
  11. Plates were read at a wavelength of 492 nanometers.

Obtained results were evaluated by comparison with standard calibrators. The optical densities of the calibrators were analyzed with the graph obtained by performing linear-log regression analysis. The minimum TNFα level that the test can measure is 5 pg/ml. was in the form.

Interleukin-6 determination: Serum IL-6 level was measured using commercially available ELISA kit (CYTELISA; The Cytoscreen, Paris, France).

  1. The samples to be studied and the kit were brought to room temperature.
  2. 50 µl of standard, patient sample and control materials were placed in each well.
  3. 50 µl of Anti-IL-6 antibody was added to them.
  4. Plates were covered and incubated for 1.5 hours at room temperature.
  5. The solutions in the wells were aspirated and emptied.
  6. The wells were filled with washing solution, waited for 15–30 seconds and aspirated. This process was repeated 3–4 times. Plates were dried by inversion.
  7. 100 µl of Streptavidin was added to all wells.
  8. Plates were closed and incubated for 30 minutes at room temperature.
  9. The steps 5 and 6 were repeated.
  10. 100 µl of coloring solution was added.
  11. Plates were sealed and incubated at room temperature for 30 minutes in the dark.
  12. Plates were read at a wavelength of 450 nanometers.

Obtained results were evaluated by comparison with standard calibrators. The optical densities of the calibrators were analyzed with the graph obtained by performing linear-log regression analysis. The minimum IL-6 level that the test can measure is 10 pg/ml. was in the form.

Interleukin-10 determination: Serum IL-10 level was measured using commercially available ELISA kit (CYTELISA; The Cytoscreen, Paris, France).

  1. The samples to be studied and the kit were brought to room temperature.
  2. 100 µl of standard, patient sample and control materials were placed in each well.
  3. 50 µl of Anti-IL-10 antibody was added to them.
  4. Plates were covered and incubated at 37°C for 2 hours.
  5. The solutions in the wells are aspirated and emptied.
  6. The wells were filled with washing solution, waited for 15–30 seconds and aspirated. This process was repeated 3–4 times. Plates were dried by inversion.
  7. 100 µl of Streptavidin was added to all wells.
  8. Plates were closed and incubated at 37°C for 45 minutes.
  9. The steps 5 and 6 were repeated.
  10. 100 µl of coloring solution was added.
  11. Plates were sealed and incubated at room temperature for 30 minutes in the dark.
  12. Plates were read at a wavelength of 450 nanometers.

Obtained results were evaluated by comparison with standard calibrators. The optical densities of the calibrators were analyzed with the graph obtained by performing linear-log regression analysis. The minimum IL-10 level that the test can measure is 5 pg/ml. was in the form.

In the statistical calculations of the numerical values obtained, the "Significance Test of the Difference Between Two Means" or the "Mann-Whitney U Test" was used in accordance with the number of cases. Results are given as arithmetic mean ± standard error. Values of P ≤ 0,05 were considered statistically significant.

Table 1 summarizes the results of the active (12 patients) BP, while Table 2 summarizes the results of the inactive (8 patients) BP. Table 3 summarizes the results of the control group.

 Lower valueUpper valueAverageStandard errorStandard deviation
Age (Year)203022,551,202,71
ESR (mm/h)2010566,256,5022,52
CRP (mg/dl)11,0047,0025,253,3611,63
Fibrinogen (mg/dl)386585474,0816,9358,64
IL-10 (pg/ml)93,40500,00257,7845,54157,76
IL-6 (pg/ml)7,8093,4032,967,1824,90
SIL-2R (pg/ml)7,80125,0034,629,2732,14
TNF-α (pg/ml)7,8062,5047,365,5319,17

Table 1: Findings of Active Behçet's Patients (n = 12)

 Lower valueUpper valueAverage

Standard error

error

Standard deviation
Age (Year)203022,601,182,68
ESR (mm/h)32012,384,518,40
CRP (mg/dl)5,907,006,030,140,38
Fibrinogen (mg/dl)265622391,5038,56109,05
IL-10 (pg/ml)15,20125,0029,8013,6238,54
IL-6 (pg/ml)7,8022,4011,452,386,75
SIL-2R (pg/ml)7,8015,6010,721,424,03
TNF-α (pg/ml)7,8042,5014,084,2412,01

Table 2: Findings of Inactive Behçet's Patients (n = 8)

 Lower valueUpper valueAverageStandard errorStandard deviation
Age (Year)202922,451,302,61
ESR (mm/h)2207,653,254,48
CRP (mg/dl)5,905,905,900,280,00
Fibrinogen (mg/dl)191382288,2021,4547,05
IL-10 (pg/ml)7,8015,608,581,382,40
IL-6 (pg/ml)7,8015,6010,141,453,66
SIL-2R (pg/ml)7,8015,609,361,403,20
TNF-α (pg/ml)15,6042,5019,638,429,85

Table 3: Findings of the control group (n = 20)

Comparison the serum levels of conventional acute phase markers of active and inactive BP with control group is showed in Figure 1.

Figure 1: Comparison of serum levels of classical acute phase markers

Comparison the serum levels of SIL2-R, IL-6, IL-10 and TNFα of active and inactive BP with control group is showed in Figure 2.

Figure 2: Comparison of serum levels of cytokines

Serum levels of ESR (P < 0,001), CRP (P < 0,001), fibrinogen (P < 0,001), IL-10 (P < 0,001), IL-6 (P < 0,001), SIL-2R (P < 0,001), and TNFα (P < 0,001) were statistically different between active and inactive Behçet’s Patients (Table 4, Fig. 1).

 ESRCRPFibrinogenIL-10IL-6SIL-2RTNF-α
Mann-Whitney U0,5000,00018,50010,50022,50014,5008,000
Wilcoxon W36,50036,0054,50046,50058,50050,50044,000
Z value-3,672-3,714-2,279-2,971-2,052-2,777-3,304
Asymp. Sig. [2- tailed]0,0000,0000,0230,0030,0170,0050,001
P value0,0000,0000,0200,0020,0470,0070,001

Table 4: Comparison of Findings of Active-Inactive Behçet's Patients

Serum levels of ESR (P < 0,001), CRP (P < 0,001), fibrinogen (P < 0,001), IL-10 (P < 0,021), IL-6 (P < 0,001), SIL-2R (P < 0,001), and TNFα (P < 0,001) were statistically different between active Behçet’s Patients and the control group (Table 5, Figure. 1).

 ESRCRPFibrinogenIL-10IL-6SIL-2RTNF-α
Mann-Whitney U0,5000,00013,5002,00024,00011,5006,000
Wilcoxon W210,500210,00225,500212,00234,00221,500216,000
Z value-4,668-4,729-4,147-5,076-4,139-4,880-4,886
Asymp. Sig. [2- tailed]0,0000,0000,0000,0000,0000,0000,000
P value0,0000,0000,0000,0000,0000,0000,000

Table 5: Comparison of Active Behçet's Patients-Control Group.

Serum levels of ESR (P = 0,746), CRP (P = 0,746), and fibrinogen (P = 0,940)) were not statistically different between inactive Behçet’s Patients and the control group (Table 6, Fig. 1).

 ESRCRPFibrinogenIL-10IL-6SIL-2RTNF-α
Mann-Whitney U73,50073,00078,0010,00020,00014,00020,000
Wilcoxon W109,500283,00114,00220,000223,000224,000223,000
Z value-0,334-0,373-0,102-4,170-3,517-4,287-4,291
Asymp. Sig. [2- tailed]0,7380,7090,9190,0000,0000,0000,000
P value0,7460,7460,9400,0000,0010,0000,001

Table 6: Comparison of Inactive Behçet's Patients-Control Group

Serum levels of IL-10 (P < 0,001), IL-6 (P = 0,001), SIL-2R (P < 0,001), and TNFα (P = 0,001) were statistically different between inactive Behçet’s patients and the control group (Fig. 2).

Discussion

The exact cause of BD is not yet known. The theory that BD is an aberrant immune response in people who are genetically predisposed to the disease that is brought on by environmental antigens or autoantigens like heat shock proteins (HSPs) is currently the one with the greatest evidence [28, 29].

Patients with BD have higher levels of proinflammatory cytokines, particularly when the disease is active. The enhanced inflammatory response is thought to be caused by this elevation, particularly those implicated in Th1-mediated inflammation, along with genetic susceptibility [28–30].

The literature is replete with studies examining IL-10 serum levels. Sadeghi et al. discovered no significant difference in serum levels of IL-10 between Behçet's patients and a control group, according to Aridogan et al. and Guenane et al [31, 32].

However, in our study, we found a substantial difference in serum IL-10 levels between patients with active BD and the control group, as well as between patients with inactive BD and those who did not have it.

Acute phase reactant CRP is known to be increased in a range of inflammatory diseases. The CRP level may potentially be a sign of tissue damage caused by the immune system. In our investigation, a substantial difference was found between the CRP levels of active BP and the control group.

Numerous studies examining CRP serum levels have been published, and several of these studies (e.g. Karada et al., Müftüolu et al., Balta et al., Bekpınar et al., Adam et al.) reveal that serum CRP levels are elevated in individuals with active BD [33–35].

In several disorders, including Behçet's patients in the active phase, investigations have shown that ESR and CRP rise as acute phase reactants [36, 37]. Most active BD symptoms are linked to both ESR and CRP. They can be applied to Behçet's patients to forecast disease activity and active vascular symptoms [37].

In addition to serving as a marker of inflammation, fibrinogen, an acute phase reactant, is essential for the production of clots in both the fibrin network and platelet aggregation. In order to understand the vascular involvement and to demonstrate the disease activity, the increase in fibrinogen in Behçet's illness is crucial [38].

These cytokines were chosen because IL-6, SIL-2R and TNFα are proinflammatory; IL-10 is known as an anti-inflammatory cytokine and there is more research and knowledge about these cytokines. In addition, the increase in serum levels of these cytokines in many inflammatory diseases suggests that they will be effective both in the pathogenesis of the disease and in the evaluation of disease activity, as well as in the treatment.

This study can be criticized in two aspects: One of them is the low number of participants, and the other is that the results were obtained in this way because there may be organ involvement in Behçet's patients in the inactive period (no real inactive period). However, it is not easy to find young male patients who have never received treatment. Yes, it would be better if more young men participated in this study. Perhaps in the future, such a study will be conducted with a large number of participants.

Secondly, it is important that although the classical acute phase markers are normal, cytokines are high in these patients, even though Behçet's patients in the inactive period are not really in the inactive period due to organ involvement. Because detecting a condition that cannot be detected with conventional acute phase markers with cytokine levels is very important for morbidity and mortality follow-up.

Another question that may come to mind: Why was IL-10, an anti-inflammatory cytokine, found to be high, as well as high levels of proinflammatory cytokines in Behçet's patients in the inactive period? In response to this: Perhaps the inactive period of these patients is due to this anti-inflammatory cytokine. IL-10, which is an anti-inflammatory cytokine, increases more because proinflammatory cytokines increase more in the active period, and IL-10 increases less in the inactive period, since the increase in proinflammatory cytokines is less.

Declarations

Acknowledgements

I would like to thank my thesis advisor, Prof. Dr. Yavuz Baykal and helping with statistics Prof. Dr. Mustafa Turan and Ankara Gülhane Military Medical Academy Internal Medicine Department nurses and Immunology and Biochemistry Laboratory staff.

Funding

No funding was received

Availability of data and materials

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Authors’ contributions

This article is derived from the specialty thesis in medicine. Therefore, the only author is Ali Osman Avcı, MD. Those who contributed during the thesis work are included in the acknowledgment section.

Ethics approval and consent to participate

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of University Gulhane Military Medical Academy (2020/27).

Patient consent for publication

The author affirms that human research participants provided informed consent for publication.

Competing interests

The authors declare that they have no competing interests.

Additional Declarations

No competing interests reported.

References

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"We recently published an article entitled “Influence of beta-Cyclodextrins upon the Degradation of Carbofuran Derivatives under Alkaline Conditions" in the Journal of “Pesticides and Biofertilizers” to show that the cyclodextrins protect the carbamates increasing their half-life time in the presence of basic conditions This will be very helpful to understand carbofuran behaviour in the analytical, agro-environmental and food areas. We greatly appreciated the interaction with the editor and the editorial team; we were particularly well accompanied during the course of the revision process, since all various steps towards publication were short and without delay".

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Jesus Simal-Gandara

I would like to express my gratitude towards you process of article review and submission. I found this to be very fair and expedient. Your follow up has been excellent. I have many publications in national and international journal and your process has been one of the best so far. Keep up the great work.

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Douglas Miyazaki

We are grateful for this opportunity to provide a glowing recommendation to the Journal of Psychiatry and Psychotherapy. We found that the editorial team were very supportive, helpful, kept us abreast of timelines and over all very professional in nature. The peer review process was rigorous, efficient and constructive that really enhanced our article submission. The experience with this journal remains one of our best ever and we look forward to providing future submissions in the near future.

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Dr Griffith

I am very pleased to serve as EBM of the journal, I hope many years of my experience in stem cells can help the journal from one way or another. As we know, stem cells hold great potential for regenerative medicine, which are mostly used to promote the repair response of diseased, dysfunctional or injured tissue using stem cells or their derivatives. I think Stem Cell Research and Therapeutics International is a great platform to publish and share the understanding towards the biology and translational or clinical application of stem cells.

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Dr Tong Ming Liu

I would like to give my testimony in the support I have got by the peer review process and to support the editorial office where they were of asset to support young author like me to be encouraged to publish their work in your respected journal and globalize and share knowledge across the globe. I really give my great gratitude to your journal and the peer review including the editorial office.

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Husain Taha Radhi

I am delighted to publish our manuscript entitled "A Perspective on Cocaine Induced Stroke - Its Mechanisms and Management" in the Journal of Neuroscience and Neurological Surgery. The peer review process, support from the editorial office, and quality of the journal are excellent. The manuscripts published are of high quality and of excellent scientific value. I recommend this journal very much to colleagues.

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S Munshi

Dr.Tania Muñoz, My experience as researcher and author of a review article in The Journal Clinical Cardiology and Interventions has been very enriching and stimulating. The editorial team is excellent, performs its work with absolute responsibility and delivery. They are proactive, dynamic and receptive to all proposals. Supporting at all times the vast universe of authors who choose them as an option for publication. The team of review specialists, members of the editorial board, are brilliant professionals, with remarkable performance in medical research and scientific methodology. Together they form a frontline team that consolidates the JCCI as a magnificent option for the publication and review of high-level medical articles and broad collective interest. I am honored to be able to share my review article and open to receive all your comments.

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Tania Munoz

“The peer review process of JPMHC is quick and effective. Authors are benefited by good and professional reviewers with huge experience in the field of psychology and mental health. The support from the editorial office is very professional. People to contact to are friendly and happy to help and assist any query authors might have. Quality of the Journal is scientific and publishes ground-breaking research on mental health that is useful for other professionals in the field”.

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George Varvatsoulias

Dear editorial department: On behalf of our team, I hereby certify the reliability and superiority of the International Journal of Clinical Case Reports and Reviews in the peer review process, editorial support, and journal quality. Firstly, the peer review process of the International Journal of Clinical Case Reports and Reviews is rigorous, fair, transparent, fast, and of high quality. The editorial department invites experts from relevant fields as anonymous reviewers to review all submitted manuscripts. These experts have rich academic backgrounds and experience, and can accurately evaluate the academic quality, originality, and suitability of manuscripts. The editorial department is committed to ensuring the rigor of the peer review process, while also making every effort to ensure a fast review cycle to meet the needs of authors and the academic community. Secondly, the editorial team of the International Journal of Clinical Case Reports and Reviews is composed of a group of senior scholars and professionals with rich experience and professional knowledge in related fields. The editorial department is committed to assisting authors in improving their manuscripts, ensuring their academic accuracy, clarity, and completeness. Editors actively collaborate with authors, providing useful suggestions and feedback to promote the improvement and development of the manuscript. We believe that the support of the editorial department is one of the key factors in ensuring the quality of the journal. Finally, the International Journal of Clinical Case Reports and Reviews is renowned for its high- quality articles and strict academic standards. The editorial department is committed to publishing innovative and academically valuable research results to promote the development and progress of related fields. The International Journal of Clinical Case Reports and Reviews is reasonably priced and ensures excellent service and quality ratio, allowing authors to obtain high-level academic publishing opportunities in an affordable manner. I hereby solemnly declare that the International Journal of Clinical Case Reports and Reviews has a high level of credibility and superiority in terms of peer review process, editorial support, reasonable fees, and journal quality. Sincerely, Rui Tao.

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Rui Tao

Clinical Cardiology and Cardiovascular Interventions I testity the covering of the peer review process, support from the editorial office, and quality of the journal.

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Khurram Arshad

Clinical Cardiology and Cardiovascular Interventions, we deeply appreciate the interest shown in our work and its publication. It has been a true pleasure to collaborate with you. The peer review process, as well as the support provided by the editorial office, have been exceptional, and the quality of the journal is very high, which was a determining factor in our decision to publish with you.

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Gomez Barriga Maria Dolores

The peer reviewers process is quick and effective, the supports from editorial office is excellent, the quality of journal is high. I would like to collabroate with Internatioanl journal of Clinical Case Reports and Reviews journal clinically in the future time.

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Lin Shaw Chin

Clinical Cardiology and Cardiovascular Interventions, I would like to express my sincerest gratitude for the trust placed in our team for the publication in your journal. It has been a true pleasure to collaborate with you on this project. I am pleased to inform you that both the peer review process and the attention from the editorial coordination have been excellent. Your team has worked with dedication and professionalism to ensure that your publication meets the highest standards of quality. We are confident that this collaboration will result in mutual success, and we are eager to see the fruits of this shared effort.

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Maria Dolores Gomez Barriga

Dear Dr. Jessica Magne, Editorial Coordinator 0f Clinical Cardiology and Cardiovascular Interventions, I hope this message finds you well. I want to express my utmost gratitude for your excellent work and for the dedication and speed in the publication process of my article titled "Navigating Innovation: Qualitative Insights on Using Technology for Health Education in Acute Coronary Syndrome Patients." I am very satisfied with the peer review process, the support from the editorial office, and the quality of the journal. I hope we can maintain our scientific relationship in the long term.

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Dr Maria Dolores Gomez Barriga

Dear Monica Gissare, - Editorial Coordinator of Nutrition and Food Processing. ¨My testimony with you is truly professional, with a positive response regarding the follow-up of the article and its review, you took into account my qualities and the importance of the topic¨.

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Dr Maria Regina Penchyna Nieto

Dear Dr. Jessica Magne, Editorial Coordinator 0f Clinical Cardiology and Cardiovascular Interventions, The review process for the article “The Handling of Anti-aggregants and Anticoagulants in the Oncologic Heart Patient Submitted to Surgery” was extremely rigorous and detailed. From the initial submission to the final acceptance, the editorial team at the “Journal of Clinical Cardiology and Cardiovascular Interventions” demonstrated a high level of professionalism and dedication. The reviewers provided constructive and detailed feedback, which was essential for improving the quality of our work. Communication was always clear and efficient, ensuring that all our questions were promptly addressed. The quality of the “Journal of Clinical Cardiology and Cardiovascular Interventions” is undeniable. It is a peer-reviewed, open-access publication dedicated exclusively to disseminating high-quality research in the field of clinical cardiology and cardiovascular interventions. The journal's impact factor is currently under evaluation, and it is indexed in reputable databases, which further reinforces its credibility and relevance in the scientific field. I highly recommend this journal to researchers looking for a reputable platform to publish their studies.

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Dr Marcelo Flavio Gomes Jardim Filho

Dear Editorial Coordinator of the Journal of Nutrition and Food Processing! "I would like to thank the Journal of Nutrition and Food Processing for including and publishing my article. The peer review process was very quick, movement and precise. The Editorial Board has done an extremely conscientious job with much help, valuable comments and advices. I find the journal very valuable from a professional point of view, thank you very much for allowing me to be part of it and I would like to participate in the future!”

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Zsuzsanna Bene

Dealing with The Journal of Neurology and Neurological Surgery was very smooth and comprehensive. The office staff took time to address my needs and the response from editors and the office was prompt and fair. I certainly hope to publish with this journal again.Their professionalism is apparent and more than satisfactory. Susan Weiner

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Dr Susan Weiner

My Testimonial Covering as fellowing: Lin-Show Chin. The peer reviewers process is quick and effective, the supports from editorial office is excellent, the quality of journal is high. I would like to collabroate with Internatioanl journal of Clinical Case Reports and Reviews.

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Lin-Show Chin

My experience publishing in Psychology and Mental Health Care was exceptional. The peer review process was rigorous and constructive, with reviewers providing valuable insights that helped enhance the quality of our work. The editorial team was highly supportive and responsive, making the submission process smooth and efficient. The journal's commitment to high standards and academic rigor makes it a respected platform for quality research. I am grateful for the opportunity to publish in such a reputable journal.

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Sonila Qirko

My experience publishing in International Journal of Clinical Case Reports and Reviews was exceptional. I Come forth to Provide a Testimonial Covering the Peer Review Process and the editorial office for the Professional and Impartial Evaluation of the Manuscript.

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Luiz Sellmann