Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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Dr Mohan Z Mani

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On Sep 2018




Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."



Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



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Professor and Head
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Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
‘Knowledge is treasure of a wise man.’ The free access of this journal provides an immense scope of learning for the both the old and the young in field of medicine and dentistry as well. The multidisciplinary nature of the journal makes it a better platform to absorb all that is being researched and developed. The publication process is systematic and professional. Online submission, publication and peer reviewing makes it a user-friendly journal.
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Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




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"My sincere attachment with JCDR as an author as well as reviewer is a learning experience . Their systematic approach in publication of article in various categories is really praiseworthy.
Their prompt and timely response to review's query and the manner in which they have set the reviewing process helps in extracting the best possible scientific writings for publication.
It's a honour and pride to be a part of the JCDR team. My very best wishes to JCDR and hope it will sparkle up above the sky as a high indexed journal in near future."



Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




Dr. C.S. Ramesh Babu
" Journal of Clinical and Diagnostic Research (JCDR) is a multi-specialty medical and dental journal publishing high quality research articles in almost all branches of medicine. The quality of printing of figures and tables is excellent and comparable to any International journal. An added advantage is nominal publication charges and monthly issue of the journal and more chances of an article being accepted for publication. Moreover being a multi-specialty journal an article concerning a particular specialty has a wider reach of readers of other related specialties also. As an author and reviewer for several years I find this Journal most suitable and highly recommend this Journal."
Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2023 | Month : January | Volume : 17 | Issue : 1 | Page : BC06 - BC10` Full Version

Assessment of Haptoglobin 2-2 Genotype in Type 2 Diabetes and Cardiovascular Patients in North Indian Population: A Case-control Study


Published: January 1, 2023 | DOI: https://doi.org/10.7860/JCDR/2023/57707.17340
Puneet, Atul Kumar Upadhyay, Manpreet Kaur, Rattan Deep Singh

1. Research Scholar, Department of Bioengineering and Biosciences, Lovely Professional University, Phagwara, Punjab, India. 2. Assistant Professor, Department of Biotechnology, Thapar Institute of Engineering and Technology, Patiala, Punjab, India. 3. Associate Professor, Department of Human Genetics, Guru Nanak Dev University, Amritsar, Punjab, India. 4. Associate Professor, Department of Bioengineering and Biosciences, Lovely Professional University, Phagwara, Punjab, India.

Correspondence Address :
Rattan Deep Singh,
Lovely Professional University, Phagwara, Phagwara, Punjab, India.
E-mail: rattan.19383@lpu.co.in

Abstract

Introduction: Haptoglobin (Hp), an acute phase protein, is a plasma inflammatory marker. It is important in both infectious and non infectious illnesses. When intravascular haemolysis occurs, the dead red blood cells are discharged into the circulation and link to Hp to form the haemoglobin (Hb)-haptoglobin (Hb-Hp) complex that is recognised by the macrophage scavenger receptor CD163. The Hp1 and Hp2 are its two alleles. The Hp2 is formed by a partial duplication of the Hp1 gene. The Hp1 has five exons, but Hp2 has seven exons owing to the duplication of Hp1 third and fourth exon. There are three genotypes of Hp: Hp1-1, Hp2-1, and Hp2-2. The encoded products of genotypes are attached to haemoglobin with varying degrees of affinity: Hp1-1 has the most affinity, Hp2-1 has medium affinity, and Hp2-2 has the lowest affinity. Hp allelic and genotypic variation differs amongst populations and different geographic regions of the world.

Aim: To study the haptoglobin 2-2 genotype in type 2 diabetes and cardiovascular patients in north Indian population.

Materials and Methods: It was a case-control study and a total of 200 participants were recruited randomly including 50 participants of type 2 Diabetes Mellitus (DM), 50 participants of Cardiovascular Disorders (CVD), 50 participants with Cardiovascular Disorders With type 2 Diabetes Mellitus (CVDWDM) and 50 normal healthy controls from Punjab Institute of Medical Sciences (PIMS) Hospital and clinics and Aashirwad laboratory of Jalandhar, Punjab, India in February 2021 to January 2022. Anthropometric variables were determined by using standard methods and biochemical parameters were measured by commercial available kits. Allele specific Polymerase Chain Reaction (PCR) or Amplification Refractory Mutation System (ARMS) PCR was used to detect Hp genotypes. Data were represented as mean±Standard Deviation and the statistical difference i.e. p<0.05 between all parameters was determined by using the Mann-Whitney U test.

Results: Anthropometric parameters i.e. Body Mass Index (BMI) was found to be statistically significant in CVDWDM as compared to CVD, N and T2DM i.e. p=0.023, p=0.01 and p=0.014. Waist Circumference (WC) was found to be statistically significant in CVDWDM as compared to CVD, N and T2DM i.e. p=0.02, p=0.013 and p=0.012 and Hip Circumference (HC) was found to be statistically significant in CVDWDM as compared to CVD, N and T2DM i.e. p=0.02, p=0.017 and p=0.014. However, Apolipoprotein A1 mean value was found to be higher and statistically significant i.e. p=0.03 in Type 2 Diabetes Mellitus (T2DM) as compared to healthy participants. Triglycerides mean value was increased in T2DM as compared to CVD, CVD as compared to N and CVDWDM as compared to healthy controls (p-value=0.012,0.04,0.015, respectively). The findings had shown that a 349 bp band was detected in patients with T2DM, CVD and CVDWDM, implying that the Hp2-2 genotypes were observed but not in healthy controls.

Conclusion: The study finds that people with DM, CVD, or CVDWDM all have a 349bp band. It means that the genotypes of the Hp2-2 allele were found in patients of diabetes, CVD and diabetics with CVD but not in the healthy individuals.

Keywords

Allele, Apolipoprotein, Glycosylated haemoglobin, Polymerase chain reaction

Diabetes is a non communicable disease that is ravaging both industrialised and developing nations. In 2011, 366 million individuals were impacted globally, and this number is expected to climb to almost 552 million by 2030 (1). Diabetes is a chronic, and life threatening metabolic illness brought on by a high blood glucose level in the body. Type 2 Diabetes Mellitus progresses slowly and is difficult to detect in its early stages (2). One of the most serious outcomes of T2DM is CVD. Similarly, throughout the last two to three decades, a CVD epidemic has emerged throughout the world. In 1990, it was estimated that 5.3 million fatalities were caused by CVD in industrialised nations, while the comparable numbers for underdeveloped nations were between 8 and 9 million (i.e. a relative excess of 70%) (3). Moreover half of those with T2DM experience a coronary artery disease, a stroke, or a heart attack (4). Hp is a glycoprotein produced in the liver in response to cytokines including IL-1, IL-6, and TNF (Tumour Necrosis Factor). Hp is a protein that looks like an immunoglobulin in which disulfide bridges connect two alpha (light) and two beta (heavy) chains (S-S). The Hp gene is found on chromosome 16q22.1, which is the long arm of the chromosome (5). The Hp1 and Hp2 are the two autosomal co dominant alleles. Hp1 has five exons, and Hp2 allele was created by the duplication or multimerisation of exons 3 and 5 of Hp1, in total there are seven exons in this gene (6),(7),(8).

In diverse ethnicities and ethnic groups across the world, there are allelic and genotypic differences in Hp. Studies have reported that in India, (Maharashtra, Rajasthan, Delhi and Uttar Pradesh) the allelic frequency of Hp was measured by using starch gel electrophoresis, polyacrylamide gel electrophoresis, and the Enzyme Linked Immunosorbent Assay (ELISA) technique from healthy people in order to conduct a caste-based study on control subjects (9),(10),(11),(12),(13),(14),(15). In India, there is no work on the molecular level related to Hp gene. For the above study, this is the first extensive genotypic analysis in Northern India. Some studies have shown that the Hp 2-2 genotype raises the possibility of coronary diseases (16),(17). The study hypothesised that CVD is substantially more likely to occur in diabetics who are homozygous for the haptoglobin 2 genotype (Hp 2-2). The impact of Hp 2-2 protein in both cardiovascular and diabetes diseases was being investigated for the first time in this work.

Material and Methods

The current case-control study included 200 people randomly (convenience sampling) and was carried out from February 2021-January 2022. Out of 200, 50 were having T2DM, 50 were CVD, 50 were CVDWDM, and 50 were healthy controls. The research work for the current study was assessed and approved by the Institutional Ethics Committee of Lovely Professional University, Phagwara, (LPU/IEC/2018/01/01). After providing their informed consent, all individuals were included from the clinic.

Inclusion criteria: The patients diagnosed with diabetes mellitus on the basis of American Diabetes Association criteria (ADA, 2015) on the basis of glycated haemoglobin (A1C) ≥6.5% and aged 26-75 years, were enrolled (males and females) from Northern India. Healthy individuals without history of any chronic disease were included in this study (18),(19).

Exclusion criteria: Patients with a history of diabetic retinopathy, gestational diabetes was excluded from the study. Pregnant ladies, nursing mothers and with age <26, and >75 years and children were excluded from the study.

Study Procedure

Anthropometric measures were obtained when the individuals were dressed in light clothes and were not wearing shoes. A calibrated stadiometer and a portable weighing machine were used to determine height and weight. A non stretchable plastic tape was used to measure the circumference of the hips. The minimal horizontal girth between the costal margins and the iliac crests at the conclusion of normal expiration was used to calculate waist circumference. Waist-Hip Ratio (WHR) was determined by waist circumference divided by hip circumference. The BMI was calculated by the weight in kilogrammes (kg) by the squared height in metres (m2) (20). Each participant's fasting venous blood (5 mL) was collected into Ethylenediamine Tetraacetic Acid (EDTA)-coated vials, and then centrifuged at 4500 rpm for 5 minutes. The resulting supernatant or serum were collected into two 1.5 mL micro-centrifuge tubes or vials and stored at -20°C for further biological investigation (21).

Biochemical Analysis

A typical glucometer gadget was used to measure Fasting Blood Glucose (FBG) levels (22). Total cholesterol, triglycerides, Apolipoprotein A1 (APOA1) and glycosylated haemoglobin (HbA1c) were measured by using conventional Quantia kits i.e. quantitative turbidimetric immunoassay technique manufactured by Coral Clinical Systems a division of Tulip Diagnostics (P) Ltd. High Density Lipoprotein (HDL) was calculated by Friedewald formula: HDL= Cholesterol/5, Very Low Density Lipoprotein (VLDL) was calculated by Friedewald formula: VLDL= Triglyceride/5 and Low Density Lipoprotein (LDL) was calculated by Friedewald formula: LDL=TC-HDL-TG/5 (22),(23).

Haptoglobin Genotyping

The inorganic technique was used to isolate genomic DNA from whole human blood samples (24). The genotyping of haptoglobin was performed using the PCR method, as published by Koch et al, with minor modifications (25). Each amplification was carried out by using 50 ng of Deoxy Ribonucleic Acid (DNA), i.e. 1μl in a volume of 20 μl involve 100 μM of primers C and D, 8 μl of PCR nuclease water and 10 μL of PCR master mix. After an initial denaturation at 94°C for 3 minutes, the PCR was run for a total of 30 cycles, with each cycle consisting of denaturation at 94°C for 30 seconds, annealing at 69°C for 30 seconds, extension at 72°C for 30 seconds, and final extension at 72°C for 7 minutes. Total 1.5% agarose gel electrophoresis was used to separate the PCR products. The amplification was found in a 349bp product that was specific to the Hp2-2 allele compared with 100 kb DNA ladder (26).

Statistical Analysis

The data was statistically evaluated with the Statistical Package for Social Science (SPSS) computer application (version 22.0). The Data were shown as a mean and Standard Deviation (SD). For abnormally distributed data, the Mann Whitney - U test was used to make comparisons between groups. Statistical significance was determined to exist when the p-value was less than 0.05.

Results

In this case-control study, 200 volunteers participated. The mean±SD and p-value were measured by Mann-Whitney U test because data were abnormally distributed. The basic demographic data of all participants as shown in (Table/Fig 1), the mean age of T2DM was (52.9±10.733), CVD was (50.92±11.761) and CVDWDM was (55.72±11.471) and in healthy controls was (45.46±13.45). Smoking, Systolic Blood Pressure (SBP), alcoholism, tobacco takers and non-takers, dietary habits and duration of T2DM, CVD and CVDWDM have been represented in percentage. (Table/Fig 2) shows the anthropometric and biochemical parameters. Among anthropometric parameters, the mean value of BMI in CVDWDM was found to be higher and statistically significant i.e. p<0.05 as compared to CVD and T2DM patients. The mean value of WC and HC in T2DM was found to be higher and statistically non-significant i.e. p>0.05 as compared to CVD. The mean value of HbA1c and FBG in T2DM was found to be lower and statistically non-significant i.e. p>0.05 as compared to CVDWDM. The lipid profile parameters i.e. TC and HDL were statistically non-significant i.e. p>0.05 in all patients as compared to healthy controls. The TG and VLDL were statistically non-significant i.e. p>0.05 in CVDWDM as compared to CVD and healthy controls. The mean value of APOA1 was found to be higher and statistically significant i.e. p<0.05 in T2DM patients as compared to healthy controls. Out of 200 participants, the high aspect of the extracted DNA utilized in PCR reaction obtained one hundred fifty 349bp Hp2-2 allele products, of which 50 were diabetics, 50 were having CVD, and 50 were cardiovascular with diabetic patients as shown in (Table/Fig 3).

Discussion

Despite major breakthroughs in diabetes prevention and medical care, CVD continues to disproportionately afflict diabetics across the world. As a result, the development of vascular problems in diabetes is clearly influenced by genetic susceptibility, and a functional variation in the Hp gene has been identified as a potential indicator of the chance of developing these issues. Early discovery of the condition and prompt treatment can lessen the disease's morbidity and death (27),(28). In our findings the anthropometric variables i.e. BMI was found to be statistically significant higher in CVDWDM as compared to CVD and T2DM patients. Similar findings were found in the research conducted by Riaz S et al. and Sheriff DS et al. Insulin resistance, the main cause of type 2 diabetes, may be contributing to the rise in BMI. Insulin sensitivity of cell membranes is significantly decreased. As a result, the crucial mechanism by which insulin helps glucose flow through the cell wall to be turned into energy is significantly hampered. As a result, extra glucose continues to circulate in the circulation, resulting in high blood sugar levels that are transferred to the liver. Once there, the sugar is transformed into fat and circulated throughout the body via the bloodstream. Obesity and weight increase are outcomes of this mechanism (23),(29).

The results of WC and HC were not found to be similar with Riaz S et al, the BMI and WHR measurements are dependent on dietary patterns and dwindling levels of physical activity. It has been hypothesised that these environmental influences, particularly in those people with metabolic genotype, reveal a hereditary vulnerability to obesity (23). The results of TC, HDL, TG and VLDL were not found to be similar with Sheriff DS et al., (29). The LDL was statistically non significant i.e. p>0.05 in T2DM as compared to healthy controls and similar finding was found in the research conducted by Sheriff DS et al. According to reports, VLDL and LDL absorption in the liver is reduced in T2DM patients, which causes their amounts of these lipoproteins in the plasma to rise, especially in the postprandial period. This condition is most frequently seen in type T2DM patients who have severe insulin insufficiency or inadequate glycemic control. Additionally, it has been stated that the lower availability of LDL receptors contributed to the restricted LDL clearance (29),(30). Previous research has connected DM and diabetic microvascular problems, such as nephropathy and retinopathy, to the Hp polymorphism, piquing researchers' interest in elucidating the function of Hp phenotypes in DM and related cardiovascular consequences (7).

The goal of this study was to gather information on Hp 2-2 and type 2 diabetes, CVD and CVDWDM in North Indian population in order to see if the Hp 2-2 are linked to the development of CVD in diabetes. This investigation revealed the existence of Hp2-2 genotype in T2DM, CVD and CVDWDM participants but not in the case of healthy participants. The role of Hp in oxidation and inflammation has sparked speculation about its possible link to vascular disease, particularly in the presence of elevated oxidative stress (16). It has been reported that diabetic people with the Hp2-2 phenotype have a five-fold higher chance of having CVD than people with the Hp1-1 phenotype. It was discovered that the Hp 2-1 phenotype was connected to an intermediate risk of CVD. Individuals without diabetes mellitus, however, did not show any connection (27). The Hp 2-2 genotype was linked to a very significant increase in the risk of severe adverse cardiac events in a study, on 935 DM patients (31). Firstly, the Hp2-2 protein molecule stays in the transmission for a longer period of time and is shown to have undergone greater oxidative damage owing to the bigger molecule of Hp1-1 (32). Secondly, in the Hp1-1 protein molecule haem variation from methemoglobin (metHb) to LDL molecule was removed but partially occurs in Hp2-2. Due to that limited haem transfer from the Hb-Hp2-2 complex to LDL causes oxidation of LDL lipids as well as protein. The lower capability of Hp2-2 protein to preserve haem molecule in the haem pocket of HbA1c (16). Hence, Hb-Hp2-2 molecules in diabetes are removed more slowly than the Hb-Hp1-1 complex. When the participants have both Hp2-2 genotype and HbA1c≥6.5%, they may be at greater risk of CVD from this increase in plasma redox-active Hb-Hp complex. This complex is further linked to HDL molecule, thus results in damage to the activity of HDL in stimulating Reverse Cholesterol Transport (RCT), decrease antioxidant activity, and increase lipoprotein oxidation from heme transport (33). The APOA1 was found statistically significant i.e. p<0.05 in T2DM patients as compared to healthy controls. ApoA-I undergoes post-translational modifications, including fatty acid acylation and oxidation, which have been linked to the environment and metabolism. Non enzymatic protein glycation and Advanced Glycation End Product production have received a lot of interest in diabetic patients. Both are linked to hyperglycemia in type 1 and type 2 diabetes, where AGE adducts bind to ApoA-I and prevent it from activating Lecithin Cholesterol Acyltransferase (LCAT), the enzyme that converts nascent HDLs to mature HDLs. The development of Coronary Artery Disease (CAD) appears to be accelerated by AGEs. Modification experiments on ApoA-I have also revealed that the most basic versions are the protein's less mature isoforms. In most forms of amyloidosis, atherosclerosis, and neurodegenerative disorders, proteolysis is assumed to play a key role. In fact, HDL-associated ApoA-I is really destroyed by macrophage metalloproteinase in coronary patients at both the N and C termini. As a result, the transition from lipid-free ApoA-I to spherical HDL particles may be hampered by the lack of the first 38 amino acids present in ApoA-I (1-38). Possibly decreasing reverse cholesterol transit and making it easier for it to attach to LDL particles. Large HDL complexes need the N-terminal amino acid residues, and their removal results in less stable HDL particles (34),(35). The Hp 2-2 genotype is assessed in patients with diabetes and related consequences. Therefore, the Hp 2-2 genotype may be a valuable indicator of a person's likelihood of developing DM and its consequences (36). The low cost and ease of use of the genotyping technique are another advantage of this work. Another application is to detect Single Nucleotide Polymorphisms (SNPs), small insertions/deletions, and copy number variations is another benefit of genotyping approaches over whole genome sequencing and genome wide association studies (which is the case for haptoglobin).

Limitation(s)

In this study, the variations between Hp1-1 and Hp1-2 individuals were not examined. The sample size was less therefore there was lack of correlation in demographic data of participants. This was a further drawback of this cohort, and future research should determine the precise sample size required for detecting possible variations among all three haptoglobin genotypes.

Conclusion

The study concluded that a 349bp band was seen in diabetes patients, cardiovascular patients, and in diabetics with CVD. It indicated that the Hp2-2 allele genotypes were discovered in these patients but not in healthy people in the population. The Hp 2-2 genotype is a CVD risk factor in type 2 diabetes patients in the North Indian population, as per this study's findings. The data showed that Hp genotype may be used to stratify patients for optimal CVD therapy in people with type 2 diabetes.

Acknowledgement

Authors would like to thank Dr. Kulbir sharma, who provided the patients samples as well as information regarding their disease history from PIMS hospital. Biochemical analysis was carried out in Aashirwad laboratory and the manuscript was drafted and reviewed by all authors' equal contribution.

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DOI and Others

DOI: 10.7860/JCDR/2023/57707.17340

Date of Submission: May 12, 2022
Date of Peer Review: Jul 16, 2022
Date of Acceptance: Sep 29, 2022
Date of Publishing: Jan 01, 2023

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? Yes
• For any images presented appropriate consent has been obtained from the subjects. NA

PLAGIARISM CHECKING METHODS:
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