Evaluating High Performance Liquid Chromatography and Red Cell Indices for Hemoglobinopathy Screening: A Retrospective Study from North India | Chapter 12 | Medical Research and Its Applications Vol. 11

 

Globally, as per The World Health Organization, seven percent of the global population are carriers of hemoglobin disorders, with thalassemia and hemoglobinopathies being the most common genetic disorders. Thalassemia and other hemoglobinopathies are also one of the major public health problems in India. As initial screening methods, Red Blood Cell (RBC) indices such as total RBC count, Mean Cell Volume (MCV), Mean Cell Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), and Red Cell Distribution Width (RDW) are valuable. High-Performance Liquid Chromatography (HPLC) has emerged as a powerful and excellent tool for the direct identification of different hemoglobin variants.

 

Aim: This study aims to utilize HPLC for identifying various hemoglobinopathies and to analyze the distribution of red blood cell (RBC) indices in different Hemoglobinopathies to assess their significance as screening tests.

 

Materials and Methods: A retrospective study was conducted on 73,333 cases from January 2017 to October 2022. EDTA samples were analyzed on an automated hematology analyzer to determine RBC indices before performing HPLC. The Bio-Rad Variant II CE-HPLC system with the Variant II-Thalassemia short program (Bio-Rad Laboratories) was used to determine different fractions of hemoglobin. Descriptive analysis was used to obtain the frequency and percentage of Hemoglobinopathies in this given Indian population. A two-tailed p-value of <0.05 was considered to be statistically significant.

 

Results: 6,242 (8.52%) cases of abnormal hemoglobin variants were recorded. Beta Thalassemia trait (5.59%) was the most prevalent hemoglobin fraction, followed by HbD Punjab heterozygous, HbE heterozygous, and Sickle cell heterozygous. RBC indices showed statistically significant differences between subjects with normal hemoglobin and those with hemoglobinopathies (p<0.0001). Significant differences in RBC indices were also found between the Beta Thalassemia trait and normal hemoglobin subjects (p<0.0001). The mean RDW value was markedly increased in both Thalassemia syndrome (36.06±6.79) and Hb-E Beta Thalassemia (32.49±6.57). The mean total RBC count was highest in Beta Thalassemia trait (5.07±0.90) and lowest in Beta Thalassemia syndrome (2.71±1.10).

 

Conclusion: RBC indices can be effectively used to screen for Thalassemia and other hemoglobinopathies as a supportive test for High-Performance Liquid Chromatography (HPLC).

 

Author(s) Details

Chakshu Bansal
Reference Lab Delhi, Metropolis Healthcare Ltd., India.

 

Geeta Chopra
Metropolis Healthcare Ltd., India.

 

Kush Kumar Singh
Reference Lab Delhi, Metropolis Healthcare Ltd., India.

 

Ranveer Singh
Reference Lab Delhi, Metropolis Healthcare Ltd., India.

 

Raj Jatale
Metropolis Healthcare Ltd., India.

 

Shibani Ramchandran
Metropolis Healthcare Ltd., India.

 

Please see the book here:- https://doi.org/10.9734/bpi/mria/v11/1611

A Comparative Study on The Efficiency of Genomic DNA Extraction Protocols for P53 Gene Polymorphism in Prostate Cancer | Chapter 8 | Microbiology and Biotechnology Research: An Overview Vol. 2

The physical, chemical, and physiological makeup of the prostate varies greatly between species. The prostate's job is to emit a milky or white fluid that is slightly alkaline and accounts for around 30% of the amount of semen in humans, together with sperm cells and seminal vesicle fluid. Prostate cancer (PC), a type of cancer that develops in the prostate gland in the male reproductive system, is one of the most prevalent cancers influencing older men in the developed world and a major inducer of mortality for elderly men. In recent decades, genetic techniques have emerged as a powerful resource across various life-related applications. DNA-based technologies, such as PCR, are increasingly utilized in research focused on demographic genetic diversity, QTL detection, marker-assisted selection, and food traceability. These methodologies necessitate extraction processes that ensure effective nucleic acid retrieval and the removal of PCR inhibitors. The initial and most critical step in molecular biology is the extraction of DNA from cells. For molecular scientists, the quality and integrity of the isolated DNA, along with the extraction method's user-friendliness and cost-effectiveness, are essential considerations. This study aimed to develop a straightforward, rapid, and cost-effective technique for extracting DNA from human peripheral blood samples, specifically comparing two normal male subjects aged 24 years (n=2) and two male patients with prostate cancer aged 65 years (n=2). The objective was to standardize a DNA extraction protocol utilizing five different extraction methods. The first method involved a modified organic approach that substituted sodium perchlorate for traditional organic solvents like phenol and chloroform, highlighting the advantages of sodium perchlorate due to its affordability and minimal storage and shipping requirements. The second method employed an enzymatic approach using proteinase K, while the third method utilized a detergent. The fourth method incorporated phenol chloroform, and the final method was based on the salting out technique. The findings indicated that the organic extraction method produces a satisfactory yield of DNA in a relatively brief period, while the enzymatic extraction method results in superior DNA purity, making it more appropriate for PCR applications. The purpose of the present study was to find a suitable procedure for DNA extraction with low cost, time, and hazards as well as high yield, purity, and excitability for PCR amplification. Among the primary strategies for obtaining DNA is from whole blood specimens, and there are a variety of protocols known to extract nucleic acids from such specimens. The five proposed methods for DNA extraction significantly lower costs by utilizing only basic and easily obtainable laboratory supplies and equipment, eliminating the need for expensive components such as K and RNase proteins. This conclusion was drawn from a comparison of five different protocols utilizing spectrophotometry, Nanodrop technology, and electrophoresis. The PCR amplification of the P53 gene using the isolated DNA was successfully achieved through these five methods. This suggests that, apart from the detergent-based method, there were no significant inhibitors present for Taq polymerase in the final solution.

 

Author (s) Details

Zaizafoon Nabeel Nasif
Department of Chemistry, College of Science, University of Mustansiriyah, Baghdad, Iraq.

Please see the book here:- https://doi.org/10.9734/bpi/mbrao/v2/5158

Malathion-Induced Changes in Gill and Intestinal Esterase Banding Patterns of Labeo rohita | Chapter 5 | Contemporary Research and Perspectives in Biological Science Vol. 5

Contamination of surface waters has been well documented worldwide and constitutes a major issue at local, regional, national, and global levels.  In fish, different insecticides can be absorbed through gills, skin or alimentary ducts. Fishes are particularly sensitive to environmental contamination of water. Hence, pollutants such as insecticides may significantly damage certain physiological and biochemical processes when they enter into the organs of fishes. The present study was undertaken to assess the toxicological effect of Malathion (an Organophosphate) on electrophoretetic banding patterns of esterase enzymes in gill and intestine tissues of freshwater fish Labeo rohita (Hamilton) at different time intervals i.e. 24H, 48H, 72H and 96H and was compared with control. The esterase isozymes were quantitatively analyzed by using 7.5٪ native polyacrylamide gel electrophoresis (PAGE) stained with α-naphthyl acetate as substrate. Three different esterase enzyme bands were detected and named Est-1; Est-2 and Est-3 with different relative mobilities such as 0.6± 0.05; 0.4± 0.05; 0.3± 0.05 in gill and intestine tissue. All three esterase bands were found in the gill and intestine tissues in control. Isoenzyme pattern exhibits differences in the various fish populations and is also used to develop a genetic sexing system. The banding pattern of esterases of different tissues has a good potential used in the identification of species. After the fish is exposed to Malathion (an Organophosphate), in gill tissue Esterase-1 showed resistance, Esterase-2 and Esterase-3 were severely affected, whereas Esterase-3 were severely affected than Esterase-2 and Esterase-1 exhibited resistance in Intestine tissue. The present study reported that the expression of tissue-specific esterase isozymes showed a differential banding pattern that could be used in toxicological studies and also could be used for the development of molecular markers for the identification of different species of fish. This present study helps in finding a proper solution for aquatic ecosystem pollution that occurs due to pesticides. Research has to continue to find alternatives for OP compounds in agriculture.

 

Author (s) Details

Venkateswara Rao Mandalapu
S.R & B.G.N.R Arts and Science College (A), Khammam, Telangana State, India.

 

Venkaiah Yanamala
Department of Zoology, Kakatiya University, Warangal-506009 T.S., India.

 

Please see the book here:- https://doi.org/10.9734/bpi/crpbs/v5/2828

Purification and Characterization of Organic Solvent Tolerant Lipase from Newly Isolated Pseudomonas aeruginosa FW_SH-1 | Chapter 3 | Novel Aspects on Chemistry and Biochemistry Vol. 3

 In the work, a novel solvent easygoing extracellular lipase from newly unique Pseudomonas aeruginosa FW_SH-1 was purified by acetone sleet and ion exchange chromatography. The purified lipase was characterized and distinguished with lipases got from other bacterial strains.  A recently isolated strain that can produce basic solvent open-minded lipase was the aim of the present study, the strain was named as FW_SH-1, that was identified as a Pseudomonas sp. The extracellular lipase from Pseudomonas aeruginosa FW_SH-1 was purified through acetone storm followed by Q-sepharose anion exchange chromatography. A ending specific exercise of 572 U/mg was achieved accompanying 26.12 fold purification and an overall improvement of 10.21%. The molecular pressure of the purified lipase was raise to be 66 kDa using SDS polyacrylamide coagulate electrophoresis. The optimum pH and hotness for the activity of the freed lipase was found expected 8.0 and 45 oC. In the presence of hydrophilic solvent like methanol, flammable liquid, and propanol, the purified lipase remained resistant. The presence of Ca2+, Mg2+, Na+, K+ and Tween 20 aroused the activity of the freed lipase.  The metal ions to a degree Ca2+, Mg2+, Na+, K+ and Tween (20 and 80) as surfactants were found to reinforce the relative activity. The basic solvents in the way that methanol, ethanol and propanol had a very little effect in deactivating the lipase.

Author(s) Details:

Chaudhry Haider Ali,
Department of Chemical Polymer and Composite Materials Engineering, University of Engineering & Technology, New Campus, KSK Lahore-54890, Pakistan and State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Abdul Sattar Qureshi,
Institute of Biotechnology and Genetic Engineering, University of Sindh, Jamshoro, Sindh, Pakistan.

Serge Maurice Mbadinga,
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Jin-Feng Liu,
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Shi-Zhong Yang,
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Bo-Zhong Mu,
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Please see the link here: https://stm.bookpi.org/NACB-V3/article/view/11022

Determination of Peanut Varieties through Chemical Tests and Electrophoresis of Soluble Seed Proteins| Chapter 8 | Research Highlights in Agricultural Sciences Vol. 4

 Variety description and identification has become perpetually significant for innocence maintenance during source production in addition to for the varietal protection under plant sort protection. Several facets of seed trade depend the ability to characterise and recognize plant varieties. Thirty-five peanut (Arachis hypogaea L.) genotypes that are standard and in the seed result chain were characterised using biochemical tools to a degree total soluble source proteins and seed and seedling traits for their reaction to NaOH, KOH, GA3, and 2,4-D tests. It was likely to categorise all of the cultivars into different groups such as sandy color and dark brown, in addition to low, moderate, and high answer of coleoptile to respective chemical compound. Each of these genotypes has a distinctive banding pattern for soluble beginning proteins on SDS-PAGE. There were 11 to 28 bands with differing molecular weights. The genotype GG 20 produced ultimate bands (28) while the genotype GG 12 produced the minute (11). The qualitative and quantitative distinctnesses between the differing genotypes were revealed by the children protein profiles of the genotypes, professed the value of electrophoretic variation in miscellaneous identification.

Author(s) Details:

P. S. Rao,
Seed Research and Technology Centre, A.N.G.R. Agricultural University, Hyderabad-500 030, India.

M. Bharathi,
Seed Research and Technology Centre, A.N.G.R. Agricultural University, Hyderabad-500 030, India.

K. Bayyapu Reddy,
Seed Research and Technology Centre, A.N.G.R. Agricultural University, Hyderabad-500 030, India.

Please see the link here: https://stm.bookpi.org/RHAS-V4/article/view/8583

Evaluating the Role of Placental Leucine Aminopeptidase (P-LAP) in Miscarriage | Chapter 5 | Current Innovations in Medicine and Medical Science Vol. 2

 The aim of this study is to determine the role of placental leucine aminopeptidase (P-LAP)  in miscarriage and to exclude direct or indirect effects of abortion by searching for the gene for this enzyme in miscarriage. (Qiagen) Total RNA is extracted from  fresh placental tissue samples using the QIAamp® RNA Tissue Mini Kit methodology. Samples can be viewed using a 0.7% Agrose gel. After RNA extraction, reverse transcribe the total RNA sample using RT-PCR (QIAGEN One Step RT-PCR Kit). Amplify the RT reaction sample (1 ml) using RT-PCR. PCR products (10 µl) per lane are separated by  electrophoresis on a 2.0% agarose gel.  During abortion, P-LAP may play a role in delaying trophoblast invasion. Collected data show that a woman undergoing a normal pregnancy (28±0.45) has her P-LAP mRNA expression in the placenta and reduced miscarriage. There is also a difference between one miscarriage and repeated miscarriages. The mean values ​​for single miscarriages (1st and 2nd) were (16.1% ± 0.84) and (13% ± 0.24), respectively, and for repeated miscarriages (1st and 2nd) were (12% ± 0.19). is. and (9% ± 0.52). P-LAP expression is significantly lower in miscarriages (P = 0.05)  and significantly lower in recurrent miscarriages than single miscarriages. The fact that there is less P-LAP mRNA in aborted placentas may suggest that p-LAP ​​function is an important component of pregnancy management.

Author(s) Details:

Taghreed H. AL Sadoon,
Department of Biochemistry, College of Medicine, University of Tikrit, Iraq.

Ferah Gh. Al-Salihi,
Department of Chemistry, College of Sciences, University of Tikrit, Iraq.

Adnan I. Al-Badran,
Department of Biology, Cell and Biotechnology Research Unit, College of Sciences, University of Basrah, Iraq.

Please see the link here: https://stm.bookpi.org/CIMMS-V2/article/view/8222

Clinical Significance of Haptoglobin, an Acute Phase Protein | Chapter 3 | Research Aspects in Biological Science Vol. 5

In the blood plasma, haptoglobin is an acute phase protein that has a high affinity for binding haemoglobin released from erythrocytes, reducing iron loss and renal injury while also inhibiting its oxidative activity. The acute phase response is modulated in a variety of ways by haptoglobin, which also possesses antibacterial properties. Since haptoglobin also has clinical relevance, it can be utilised to identify a variety of medical disorders. Humans can have one of three different haptoglobin phenotypes: Hp 1-1, Hp 2-1, or Hp 2-2. The Haptoglobin groups can be used in forensic research in cases of disputed paternity. We are all aware of how crucial it is to get trustworthy data for forensic science and study. Any inference made from a wealth of available evidence is trustworthy and unchallengeable. The gene frequency of these three haptoglobin phenotypes has been the subject of population studies all throughout the world. The study is an effort in this regard; it compares newly available data to changes in the percentage of distinct Haptoglobin types.

 

Author (s) Details

K. J. Joshi

Forensic Science Laboratory, Kalina, Mumbai, India.

B. T. Shah

Forensic Science Laboratory, Kalina, Mumbai, India.

V. R. Rathod

Forensic Science Laboratory, Kalina, Mumbai, India.

 

View Book :-  https://stm.bookpi.org/RABS-V5/article/view/7612

 

Keywords -  Electrophoresis, genotype, gene frequency, haptoglobin, page, phenotype, population study

Electrophoresis in Pharamaceutical Analysis-A Review on Recent Application | Chapter 3 | Current Approaches in Science and Technology Research Vol. 8

 Electrophoresis is a successful analytical method for separation, but it is also extremely sensitive to minor differences in molecular charge and analyte size. Because there is such a wide range of observation methods accessible, it is one of the most valuable strategies. This paper focuses on a quick overview of electrophoresis types, including capillary electrophoresis and CE-based separation modes, as well as their benefits and drawbacks, as well as their applications. This publication also provides several capillary electrophoresis-based miscellaneous electrophoretic methods [CE- ICP-MS, CE-NMR].

Author (s) Details

M. V. Kumudhavalli
Vinayaka Mission’s College of Pharmacy, Vinayaka Mission’s Research Foundation, (Deemed University) Salem, Tamilnadu, India.

S. Gayathri
Vinayaka Mission’s College of Pharmacy, Vinayaka Mission’s Research Foundation, (Deemed University) Salem, Tamilnadu, India.

V. Nandhinipriya
Vinayaka Mission’s College of Pharmacy, Vinayaka Mission’s Research Foundation, (Deemed University) Salem, Tamilnadu, India.

B. S. Venkateswarulu
Vinayaka Mission’s College of Pharmacy, Vinayaka Mission’s Research Foundation, (Deemed University) Salem, Tamilnadu, India.

M. Kumar
Vinayaka Mission’s College of Pharmacy, Vinayaka Mission’s Research Foundation, (Deemed University) Salem, Tamilnadu, India.

View Book :- https://stm.bookpi.org/CASTR-V8/article/view/1944