AUCTORES
Globalize your Research
Case Report
*Corresponding Author: Zohreh Saltanatpour, PhD, Pediatric Cell and Gene Therapy Research Center, Gene, Cell & Tissue Research Institute, Tehran University of Medical Sciences, Tehran, Iran.
Citation: M Saremi, L Saremi, S Babaniamansour, R Nazari, Z Saltanatpour. (2021) Molecular Analysis of KPC and GES Beta-Lactamase Genes in Clinical Isolates of Klebsiella pneumoniae by PCR Method. International Journal of Clinical Case Reports and Reviews. 8(2); DOI:10.31579/2690-4861/149
Copyright: © 2021 Zohreh Saltanatpour, 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: 17 June 2021 | Accepted: 17 August 2021 | Published: 23 August 2021
Keywords: klebsiella pneumonia; β-lactamases; KPC; GES
Background: Carbapenem-resistant Klebsiella pneumonia (K. pneumoniae) has been recently identified as the major class of pathogens and the treatment became the biggest challenge in this bacterium. We assessed the antibiotic resistance patterns of K. pneumoniae, the frequency of resistant strains to imipenem, meropenem, and ertapenem, and the frequency of K. pneumoniae carbapenemases (KPC), and Guiana-Extended-Spectrum (GES) metallo-β-lactamase genes.
Methods: The phenotypes of 200 strains of K. pneumonia, collected from 650 clinical samples, were isolating and identified in Qom, Iran. The antibiotic resistance pattern of the strains was analyzed against different antibiotics. The imipenem, meropenem, and ertapenem-resistant strains, were isolated and the frequency of KPC and GES genes were evaluated.
Results: K. pneumoniae strains had different resistance patterns against various antibiotics. The isolated strains with the highest and lowest antibiotic resistance were related to ampicillin, and meropenem, respectively. Investigation of the KPC and GES β lactamase showed that none of 48 imipenem, meropenem, and ertapenem-resistance isolates had the KPC gene. In addition, the GES gene was detected in 3(2.7%) of 110 ceftazidime-resistant specimens.
Conclusions: Carbapenem is useful in the treatment of K. pneumoniae infections. Due to the importance of the mechanisms of resistance is by β-lactamase genes, investigating the prevalence of β-lactamase genes can help to increase the necessity of choosing the appropriate treatment for K. pneumoniae.
Carbapenemase-producing Klebsiella pneumonia causes a wide range of infections, including types of pneumonia, urinary tract infections, liver abscesses. K. pneumoniae carbapenemases (KPC) are β-lactamases produced by Gram-negative bacteria. They efficiently hydrolyze all cephalosporins, penicillins, monobactams, β-lactamase inhibitors, and carbapenems. Since the first reports, KPC enzymes have spread across the world, although the expansion varied by geographical locations [1, 2].
The high prevalence of broad-spectrum β-lactamases in K. pneumoniae isolated from clinical specimens of patients led to the spread of antibiotic resistance while antibiotics in the β-lactam group account for the high proportion of antibiotics used in medicine. The emerging antibiotic resistance caused an important challenge in the management of K. pneumonia-related diseases [3-5].
The fluoroquinolones, trimethoprim-sulfamethoxazole, and cephalosporins are mostly used for the treatment of K. pneumoniae infections, and resistance to these antibiotics caused the treatment problems and subsequently increased mortality. Resistance bacteria to β-lactam antibiotics obtains through the mutation in chromosomal genes or the transfer of plasmids. β-lactamase is an enzyme non-covalently links to the antibiotics containing the β-lactam ring and finally inactivates them [6-8].
Carbapenems such as meropenem, imipenem, and ertapenem are resistant to many β-lactamases, as well as class 1 chromosome β-lactamases (which destroy third-generation cephalosporins). β-lactamases can decompose and hydrolyze β-lactam antibiotics and cannot be inhibited by β-lactamase inhibitors [9, 10].
In K. pneumoniae, various groups of β-lactamases have been identified, such as KPC and GES (Guiana-Extended-Spectrum). These enzymes are usually transferred by integrons. The ability to transfer and hydrolyze the carbapenems made the antibiotics ineffective [11, 12]
KPC hydrolyses the β-lactam drugs, including monobactam, carbapenem, and third-generation falosporin. The producer gene is located on the plasmid and can transfer to many bacteria [13]. GES was first discovered in 2000 in a clinical specimen of K. pneumoniae isolated from Guiana, France. Nine enzymes of GES type have been identified in different species of pathogens such as Pseudomonas aeruginosa. GES-1 has the same hydrolytic activity as an extended-spectrum β-lactamase and is inhibited by β-lactamase inhibitors. GES-2, GES-4, GES-5, and GES-6 enzymes have also hydrolytic activity against carbapenems. GES-1, GES-5, GES-8, and GES-9 enzymes were reported in Pseudomonas aeruginosa [14-16].
Following the growing population of gram-negative organisms producing β-lactamases, there are numerous reports of widespread outbreaks, especially in the intensive care unit. The Center for Disease Control and Prevention has proposed the Polymerase chain reaction (PCR) and Real-Time PCR tests and sequencing to the microbiologists to identify these resistance producer genes, and in 2011, many scholars have used these methods. Herein, this study aimed to assess the antibiotic resistance patterns of K. pneumoniae, the frequency of resistant strains to meropenem, imipenem, and ertapenem, as well as the frequency of KPC and GES metallo-β-lactamase genes.
Study design
This study was conducted among the patients hospitalized in various wards and different hospitals of Qom city, Iran, during October 2016 and September 2017. The executive protocol of the study was approved by the ethics committee of Qom Islamic Azad University of Medical Sciences approved the executive protocol of the study. The study was conducted under the Declaration of Helsinki (seventh revision 2013). The written consent was taken from all participants before entering the study and under the direction of the Iranian Foundation of Martyrs and Veterans Affairs review board. This study did not impose any additional costs on the patients or the health system.
Sampling
The participants were asked to fill out the demographic questionnaire including gender, age, duration and ward of hospitalization, type of disease, and type of clinical specimen. Samples were taken from blood, urine, burn wounds, surgical wounds, respiratory secretions, cerebrospinal fluid, clones, and general secretions (ears, nose, and eyes). Samples were immediately transferred to the research laboratory of Qom Islamic Azad University. The samples were cultured in a MacConkey agar with a selective and differential medium. In this environment, bile salts and violet crystals inhibit the growth of gram-positive bacteria that are used for this purpose. Then the colonies, morphologically similar to K. pneumoniae, were selected and used for further investigations.
Isolation and identification of K. pneumoniae
A few bacterial colonies were removed from the solid medium, were placed along with a drop of the physiological serum on a clean slide, and were fixed by spreading and drying. Then the crystal violet was poured onto the slide and was washed with water for one minute. In the next step, the extension was adjacent to the logo for one minute and after washing the slide with water, the de-staining step was carried out with alcohol. After washing with water, slides were stained with fuchsin for 30 seconds and then thoroughly washed and dried. Finally, the slides were examined to detect the presence of gram-negative bacilli by optical microscopy. The colonies in the gram-negative bacterial staining were streaking on the agar media several times, to be completely pure. After gram staining on single-cell colonies and ensuring their purity, differential tests were performed to detect K. pneumoniae as following:
Triple Sugar Iron (TSI) test
The TSI test is used to determine whether gram-negative bacteria can to utilize lactose, glucose, sucrose and also used to generate hydrogen sulfide. After the deep and superficial cultivation of the bacteria in the TSI environment, the culture medium is incubated at 37 ° C for 24 hours.
Catalase test
The catalase enzymes mediate the breakdown of hydrogen peroxide into water and oxygen. A small amount of bacterial colony is transferred to the surface of a clean and dry glass slide using a loop. A drop of 3% H2O2 is added and placed on the glass slide. The positive test defines as the appearance of oxygen gas bubbles on the glass slide.
Citrate test
The citrate test is applied to determine the ability of K. pneumoniae to use ammonium salt as a source of nitrogen and sodium citrate as a source of carbon. K. pneumoniae that grows in the medium changes the medium alkaline. A pure colony was removed from the media and diagonally cultured on a tube containing the citrate. The appearance of a dark blue color after 24 to 48 hours at 37 ° C indicates the consumption of citrate by the organism and the positive test. Moreover, the Methyl Red and Voges-Proskauer and Sulfiter Indole Motility tests were performed based on the study conducted by Yumi et al. [17].
Oxidase test
The oxidase test is used to identify the bacteria that produce cytochrome c oxidase. The cytochrome c oxidase oxidizes the reagent (tetramethyl-p-phenylenediamine) to (indophenols) purple color end product. Therefore, one drop of reagent oxidase is placed on a filter paper, and some bacterial colonies are placed on it. After a few seconds, the absence of color change represents the negative test.
Antibiotic sensitivity test
According to the Clinical and Laboratory Standards Institute guidelines, the antibiotic resistance of the strains against selected antibiotics is evaluated by a Kirby-Bauer disc method. (Table 1).
The phenotypic identification of β-lactamase strains
The Double Disk Synergy Test was used to isolate β-lactamase producing strains. 5 ml of the ethylenediaminetetraacetic acid (EDTA) 0.5-mole solution is added to the imipenem disks. It is incubated at 4°C for 12 weeks after the drying. Imipenem-resistant strains were cultured on the Muller Hinton medium. Then, an imipenem disk (10μg) containing 930 μg EDTA was placed at an appropriate distance on the plate surface and incubated at 37 °C for 18 to 16 hours. An increase in the diameter of inhibition zone diameter, ≤ 7 mm around the imipenem disk containing EDTA, is compared to the imipenem disk alone. The results indicated the production of Metallo-β-lactamase.
Molecular analysis of β-lactamase genes in K. pneumoniae strains isolated by PCR method
PCR method was used for molecular detection of β-lactamase genes in K. pneumoniae isolated from clinical specimens. In this study, two genes of KPC and GES were selected for molecular analysis. The sequences of these primers were studied in numerous papers and their thermodynamic properties were evaluated using Gene Runner software and synthesized (Table 2).
Bacterial deoxyribonucleic acid (DNA) is extracted according to the boiling method. Amplification conditions for KPC reactions were as follows: one cycle at 94 °C for 3 minutes, 36 cycles at 94 °C for 60 seconds, 45 °C for 60 seconds and 72 °C for 60 seconds, followed by a final extension at 72 °C for 10 minutes. Amplification conditions for GES reactions were as follows: one cycle at 94 °C for 5 minutes, 36 cycles at 94 °C for 30 seconds, 45 °C for 40 seconds and 72 °C for 50 seconds, followed by a final extension at 72 °C for 5 minutes [18].
The results of isolated K. pneumoniae
After culturing the samples in the MacConkey Agar medium, those colonies with a more similar appearance to K. pneumoniae which were defined as gram-negative bacteria in the straining are considered as K. pneumoniae in the first stage. In the next step, after performing catalase and oxidase tests, the samples with positive catalase and negative oxidase tests were selected. The specimens that had resulted in differential environments, according to table 3, were identified as. K. pneumoniae
From 200 K. pneumoniae strains that were collected and confirmed by biochemical method, urine and cerebral fluid were the most and the least frequent samples, respectively. (Table 4).
Antibiotic resistance pattern
After isolating and detecting 200 samples of K. pneumoniae from clinical specimens, the resistance pattern of these strains was investigated using the disc diffusion method according to the Clinical and Laboratory Standards Institute's criteria against different antibiotics. The antimicrobial resistance pattern of K. pneumoniae isolates showed that the highest antibiotic resistance was against ticarcillin-clavulanic acid and the least resistance related to tobramycin. (Table 5).
Of the 200 strains examined, 48 strains (24%) were resistant to imipenem, ertapenem, and meropenem. These strains were selected and subsequently used for phenotypic testing of β-lactamase production and molecular analysis of β-lactamase genes.
Phenotypic test for β-lactamase
This test is carried out to evaluate the phenotypic characteristics of β-lactamase producing strains. Among 48 strains resistant to imipenem, meropenem, and ertapenem, 15 strains (31.25%) showed the phenotype of β-lactamase. (Figure 1).
The minimum inhibitory concentration (MIC) of imipenem performed on K. pneumoniae strains resistance to imipenem, were as following: 4 strains (1.92%) MIC 4µg/L, 5 strains (2.4%) MIC 8µg/L, 9 strains (2.06%) MIC 16 µg/L, 2 strains (0.95%) MIC 32µg/L, 3 strains (1.44%) MIC 64µg/L, 2 strains (0.95%) MIC 128 µg/L, 3 strains (1.44%) MIC 256 µg/L, 5 strains (2.44%) MIC 512µg/L. (Figure 2).
Amplification of the GES gene
After extraction of DNA from imipenem-resistant strains, PCR was performed for the GES gene. The results of the PCR reaction using GES primers on imipenem-resistant strains showed that of 110 strains of ceftazidime-resistant K. pneumoniae, three strains had the GES gene and subsequently 863 bp band in the PCR reaction. (Figure 3).
Amplification of the KPC gene
The results of the PCR reaction using KPC primers on imipenem-resistant strains showed that of 48 strains of imipenem-resistant K. pneumonia, none had the KPC.
K. pneumoniae has been widely distributed in hospitals and known as an opportunistic bacterium for developing various types of nosocomial infections in hospitalized patients [19, 35, 36]. Antimicrobial resistance complicated the treatment of infections related to K. pneumoniae. The carbapenems are mostly the last line treatment for K. pneumoniae infections and the emerging resistance to carbapenems caused a lot of concerns for the physicians [20, 21]. Carbapenems- resistance in K. pneumoniae is linked to various mechanisms, and the production of certain β-lactamases possessing even with a weak carbapenemase activity might lead to carbapenems resistance. True carbapenemases belong to ambler molecular class A such as GES and KPC, class B such as Verona integrin encoded metallo-β-lactamase (VIM), New Dehli metallo-β-lactamase and Imipenemase (IMP) or class D such as OXA-48-like and considered as the main agent for resistance to carbapenems without additional permeability defects in K. pneumoniae [22-24].
We studied the antibiotic resistance patterns and the frequency of β-lactamase genes (KPC and GES) in strains isolated from hospitals of Qom, Iran. The results showed that among 110 strains resistant to ceftazidime, 3 strains were positive for the GES gene, and the imipenem-resistance isolates (48 samples) none of them had the KPC gene. The most effective aminoglycoside was tobramycin. The most effective quinolone was ciprofloxacin. The most effective β-lactam was imipenem. In other studies, with the relative increase in resistance, ciprofloxacin and its derivatives have always been the most effective quinolone, amikacin the most effective aminoglycoside, and imipenem the most effective β-lactam in the treatment of Pseudomonas infections.
In studies by Amin et al. in Pakistan [25] , Sultan Dallal et al. in Tehran [26], Ishii in Japan [27] and AL-Shora et al. in Jordan [28] imipenem is introduced as an effective antibiotic for the treatment of K. pneumoniae which was consistent with our results.
The presence of broad-spectrum β-lactamases on the transposons and their probability of movement on the genome may simultaneously express the genes' resistance. This may also result in the genes' resistance to other antibiotics of broad-spectrum cephalosporins such as the GES gene and carbapenems. The GES gene was reported by Poirel et al. in the blood culture of a patient with cerebral malaria and infectious pneumonia in a general hospital in South Africa, for the first time [29] and then in 2004 by Pastrana et al, in Argentina. Some studies were assessed the molecular characteristics and methods for GES identification. The presence of GES reduces the sensitivity to imipenem. Thus, imipenem-resistant strains are also tested for this gene [30, 31].
In Japan, all strains resistant to carbapenem have been evaluated every year and only two types of metallo-β-lactamases including VIM and IMP were reported, yet. Studies showed that the prevalence of VIM in Iran was lower than that of other countries such as South Korea, Japan, and France, and probably other resistance mechanisms such as efflux pumps, defects, and lack of external membrane proteins, including outer membrane protein D and carbapenem class D are involved in producing resistance to carbapenems [32-35].
Long-term antibiotic uses enhance the generation of the organisms' resistance to the intended or other antibiotics. Paying attention to antibiotic susceptibility testing (AST), whether in terms of testing and reporting the results or in terms of considering the reported results at the time of prescribing and treating the disease, is of great value. Several preventive methods can use to control the prevalence of resistant organisms such as avoiding improper prescriptions, choosing the right antibiotic in the treatment phase, avoiding the arbitrary use of antibiotics.
Leila Saremi, Sepideh Babaniamansour, and Marzieh Saremi were in charge of samples collection, experimental design and PCR. Razieh Nazari and Marzieh Saremi contributed equally performed and designed experiments, analyzed data, and generated figures. Razieh Nazari and Zohreh Saltanatpour were in charge of idea and concept of the paper. Leila Saremi, Zohreh Saltanatpour, and Sepideh Babaniamansour assisted in supervision. All authors read and approved the final manuscript.
The authors would like to thank the Qom Islamic Azad University of Medical Sciences for their contribution to this research.
Funding: No Financial support has been received.
Ethic approval statement: The implementation of the project was approved by the ethics committee of Qom Islamic Azad University of Medical Sciences.
Conflict of interests: None.
Availability of data and material: Data sharing is available upon the author's requests.
Patient consent statement: Informed consent was obtained from all the participants prior to the study.
Consent for publication: Not applicable.
Code availability: Not applicable.
Clearly Auctoresonline and particularly Psychology and Mental Health Care Journal is dedicated to improving health care services for individuals and populations. The editorial boards' ability to efficiently recognize and share the global importance of health literacy with a variety of stakeholders. Auctoresonline publishing platform can be used to facilitate of optimal client-based services and should be added to health care professionals' repertoire of evidence-based health care resources.
Journal of Clinical Cardiology and Cardiovascular Intervention The submission and review process was adequate. However I think that the publication total value should have been enlightened in early fases. Thank you for all.
Journal of Women Health Care and Issues By the present mail, I want to say thank to you and tour colleagues for facilitating my published article. Specially thank you for the peer review process, support from the editorial office. I appreciate positively the quality of your journal.
Journal of Clinical Research and Reports I would be very delighted to submit my testimonial regarding the reviewer board and the editorial office. The reviewer board were accurate and helpful regarding any modifications for my manuscript. And the editorial office were very helpful and supportive in contacting and monitoring with any update and offering help. It was my pleasure to contribute with your promising Journal and I am looking forward for more collaboration.
We would like to thank the Journal of Thoracic Disease and Cardiothoracic Surgery because of the services they provided us for our articles. The peer-review process was done in a very excellent time manner, and the opinions of the reviewers helped us to improve our manuscript further. The editorial office had an outstanding correspondence with us and guided us in many ways. During a hard time of the pandemic that is affecting every one of us tremendously, the editorial office helped us make everything easier for publishing scientific work. Hope for a more scientific relationship with your Journal.
The peer-review process which consisted high quality queries on the paper. I did answer six reviewers’ questions and comments before the paper was accepted. The support from the editorial office is excellent.
Journal of Neuroscience and Neurological Surgery. I had the experience of publishing a research article recently. The whole process was simple from submission to publication. The reviewers made specific and valuable recommendations and corrections that improved the quality of my publication. I strongly recommend this Journal.
Dr. Katarzyna Byczkowska My testimonial covering: "The peer review process is quick and effective. The support from the editorial office is very professional and friendly. Quality of the Clinical Cardiology and Cardiovascular Interventions is scientific and publishes ground-breaking research on cardiology that is useful for other professionals in the field.
Thank you most sincerely, with regard to the support you have given in relation to the reviewing process and the processing of my article entitled "Large Cell Neuroendocrine Carcinoma of The Prostate Gland: A Review and Update" for publication in your esteemed Journal, Journal of Cancer Research and Cellular Therapeutics". The editorial team has been very supportive.
Testimony of Journal of Clinical Otorhinolaryngology: work with your Reviews has been a educational and constructive experience. The editorial office were very helpful and supportive. It was a pleasure to contribute to your Journal.
Dr. Bernard Terkimbi Utoo, I am happy to publish my scientific work in Journal of Women Health Care and Issues (JWHCI). The manuscript submission was seamless and peer review process was top notch. I was amazed that 4 reviewers worked on the manuscript which made it a highly technical, standard and excellent quality paper. I appreciate the format and consideration for the APC as well as the speed of publication. It is my pleasure to continue with this scientific relationship with the esteem JWHCI.
This is an acknowledgment for peer reviewers, editorial board of Journal of Clinical Research and Reports. They show a lot of consideration for us as publishers for our research article “Evaluation of the different factors associated with side effects of COVID-19 vaccination on medical students, Mutah university, Al-Karak, Jordan”, in a very professional and easy way. This journal is one of outstanding medical journal.
Dear Hao Jiang, to Journal of Nutrition and Food Processing We greatly appreciate the efficient, professional and rapid processing of our paper by your team. If there is anything else we should do, please do not hesitate to let us know. On behalf of my co-authors, we would like to express our great appreciation to editor and reviewers.
As an author who has recently published in the journal "Brain and Neurological Disorders". I am delighted to provide a testimonial on the peer review process, editorial office support, and the overall quality of the journal. The peer review process at Brain and Neurological Disorders is rigorous and meticulous, ensuring that only high-quality, evidence-based research is published. The reviewers are experts in their fields, and their comments and suggestions were constructive and helped improve the quality of my manuscript. The review process was timely and efficient, with clear communication from the editorial office at each stage. The support from the editorial office was exceptional throughout the entire process. The editorial staff was responsive, professional, and always willing to help. They provided valuable guidance on formatting, structure, and ethical considerations, making the submission process seamless. Moreover, they kept me informed about the status of my manuscript and provided timely updates, which made the process less stressful. The journal Brain and Neurological Disorders is of the highest quality, with a strong focus on publishing cutting-edge research in the field of neurology. The articles published in this journal are well-researched, rigorously peer-reviewed, and written by experts in the field. The journal maintains high standards, ensuring that readers are provided with the most up-to-date and reliable information on brain and neurological disorders. In conclusion, I had a wonderful experience publishing in Brain and Neurological Disorders. The peer review process was thorough, the editorial office provided exceptional support, and the journal's quality is second to none. I would highly recommend this journal to any researcher working in the field of neurology and brain disorders.
Dear Agrippa Hilda, Journal of Neuroscience and Neurological Surgery, Editorial Coordinator, I trust this message finds you well. I want to extend my appreciation for considering my article for publication in your esteemed journal. I am pleased to provide a testimonial regarding the peer review process and the support received from your editorial office. The peer review process for my paper was carried out in a highly professional and thorough manner. The feedback and comments provided by the authors were constructive and very useful in improving the quality of the manuscript. This rigorous assessment process undoubtedly contributes to the high standards maintained by your journal.
International Journal of Clinical Case Reports and Reviews. I strongly recommend to consider submitting your work to this high-quality journal. The support and availability of the Editorial staff is outstanding and the review process was both efficient and rigorous.
Thank you very much for publishing my Research Article titled “Comparing Treatment Outcome Of Allergic Rhinitis Patients After Using Fluticasone Nasal Spray And Nasal Douching" in the Journal of Clinical Otorhinolaryngology. As Medical Professionals we are immensely benefited from study of various informative Articles and Papers published in this high quality Journal. I look forward to enriching my knowledge by regular study of the Journal and contribute my future work in the field of ENT through the Journal for use by the medical fraternity. The support from the Editorial office was excellent and very prompt. I also welcome the comments received from the readers of my Research Article.
Dear Erica Kelsey, Editorial Coordinator of Cancer Research and Cellular Therapeutics Our team is very satisfied with the processing of our paper by your journal. That was fast, efficient, rigorous, but without unnecessary complications. We appreciated the very short time between the submission of the paper and its publication on line on your site.
I am very glad to say that the peer review process is very successful and fast and support from the Editorial Office. Therefore, I would like to continue our scientific relationship for a long time. And I especially thank you for your kindly attention towards my article. Have a good day!
"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".
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.
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.
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.
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.
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.
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.
“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”.
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.
Clinical Cardiology and Cardiovascular Interventions I testity the covering of the peer review process, support from the editorial office, and quality of the journal.
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.
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.
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.
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.
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¨.
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.
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!”
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
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.
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.
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.
