In vitro and ex vivo Studies of Nonionic Surfactant Vesicles Using 23 Factorial Design: Metoprolol Tartrate Used as Model Drug | Chapter 04 | Modern Advances in Pharmaceutical Research Vol. 2

The purpose of this study was to formulate and investigate metoprolol tartrate (MT) loaded non-ionic surfactant vesicles using 23 factorial designs. Preparation of niosomal drug delivery of MT increased its bioavailability which led to being better therapeutic effects, reduced the frequency of dosing and decreased side effects of hypertensive patients. Ether injection method (EIM) and thin film hydration method (TFHM) were used for the preparation of all formulations as per full factorial design to study the effect of two independent variables X1 (amount of span-60), and X2 (amount of cholesterol) on three dependent variable Y1 (percent drug entrapment efficiency), Y2 (percent drug content) and Y3 (percent cumulative drug release) respectively. The relation between the dependent and independent variables was drawn out from the mathematical equation and response surface methodology (RSM). Statistical analysis was performed using ANOVA. Microscopic observation confirmed that all particles were uniform in size and shape. The particle size of niosomes measured by SEM was between 3 μm to 4.5 μm that given the evidence of large uni-lamellar vesicles formed by EIM and TFHM. The percent drug entrapment efficiency was found to be highest for formulations MTEIM-8 and MTTFHM-8 with values 97.11% and 95.56% respectively. In vitro dissolution studies were carried out in phosphate buffer (pH 6.8) for 8 hours at 100 rpm and maintained at 37 ± 0.5°C according to USP-II paddle method and absorbance was taken at 226 nm. The probable drug release mechanism may be fickian (class I) diffusion as the correlation coefficient (𝑅2) best fitted with zero order and release exponent (n) was less than 0.43.  The FTIR studies have been done to confirm no interaction along with drug and polymer. In vitro and ex vivo comparative studies showed that non-ionic surfactant vesicle had controlled the release of drug for a longer period. Finally, it can be concluded that non-ionic surfactant vesicle could be an effective for delivery of MT with increased bioavailability.

Author(s) Details

Dr. Irin Dewan
Pharmaceutical Technology Research Laboratory, Department of Pharmacy, University of Asia Pacific, 74/A, Green Road, Farmgate, Dhaka – 1215, Bangladesh.

Prof. Dr. S. M. Ashraful Islam
Pharmaceutical Technology Research Laboratory, Department of Pharmacy, University of Asia Pacific, 74/A, Green Road, Farmgate, Dhaka – 1215, Bangladesh.

Prof. Dr. Swarnali Islam Khandaker
Pharmaceutical Technology Research Laboratory, Department of Pharmacy, University of Asia Pacific, 74/A, Green Road, Farmgate, Dhaka – 1215, Bangladesh.

Waheeda Nasreen
Pharmaceutical Technology Research Laboratory, Department of Pharmacy, University of Asia Pacific, 74/A, Green Road, Farmgate, Dhaka – 1215, Bangladesh.

Ohinul Hoque
Pharmaceutical Technology Research Laboratory, Department of Pharmacy, University of Asia Pacific, 74/A, Green Road, Farmgate, Dhaka – 1215, Bangladesh.

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The Future of Prodrugs Design | Chapter 03 | Modern Advances in Pharmaceutical Research Vol. 2

When knowledge fails to provide answers to important questions such as how to improve the bioavailability of vital medications, “Imagination is more important than knowledge,” as Albert Einstein once said. Ingenuity in the design of effective prodrugs has been lacking in quantity and quality. The reasons behind the low quality of ingenuity could be related to the fact that medicinal chemists have expertise in organic and organometalic chemistry not in biochemistry and biology. On the other hand, pharmaceutical chemists, biologists and biochemists do not have the expertise to make sophisticated chemical devices. Therefore, in order for a prodrug strategy to work, a team consisting of all this expertise is necessary.

Author(s) Details

Professor Rafik Karaman
Department of Bioorganic Chemistry, Faculty of Pharmacy, Al-Quds University, P.O. Box 20002, Jerusalem, Palestine and
Department of Sciences, University of Basilicata, Viadell’Ateneo Lucano 10, 85100, Potenza, Italy.

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Perceptions of the Traditional Medical Practitioners of North-Western Nigeria on Malaria Treatment and the Potential Antiplasmodial Properties of Plumeria rubra Stem-Bark | Chapter 02 | Modern Advances in Pharmaceutical Research Vol. 2

Aims: The apparent lack of scientific proof of efficacies claimed by the traditional medical practitioners (TMPs) (locally known as Magori/’Yan-ganye, in Hausa language) of North-Western Nigeria with respect to malaria and the many drawbacks of the current antimalarial drugs stimulated this study. The study was carried out to evaluate the perception of the TMPs on the causes, diagnosis and treatment of malaria and evaluate the potential antiplasmodial properties (in-vivo in Albino mice) of Plumeria rubra Linn. (Apocynaceae) commonly used in traditional treatment of malaria in North-Western Nigeria. The study was aimed at providing scientific basis for use of traditional health knowledge and use of medicinal plant resources in the treatment of malaria.

Study Design: Using an ethno-medical survey, information was obtained from the TMPs relating to identification of plants, their medicinal uses and the mode of preparations of remedies on traditional treatment of malaria.

Place and Duration of Study: The ethno-medicinal survey was carried out at the premises of TMPs from December, 2005 to May, 2008. The laboratory work was carried out at the Department of Pharmacognosy and Drug Development, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria from July, 2008 to February, 2010.

Methodology: An ethno-medical survey was conducted in twenty Local Government Areas across four States (Kaduna, Kano, Katsina and Jigawa) in North-Western Nigeria. The communities covered in the survey were selected on the basis of their reputation for being homes to a number of TMPs. The plant used was selected on the basis of ethno-medical information obtained from the TMPs using an exclusion criterion based on claim for activity score. The preferable solvent used by the local people was found to be mostly water and/or alcohol, the plant material was therefore extracted by maceration technique using 70% v/v aqueous-ethanol. The metabolites profiles of the extracts were determined using thin layer chromatographic (TLC) technique on commercially prepared silica gel pre-coated flexible plates.

Results: The TMPs were able to define, diagnose and presumably treat malaria using the indigenous medicines. Median lethal dose (LD50) was established to be greater than 5 gkg-1 for the aqueous extract and 3.8 gkg-1 for the chloroform extract orally in mice respectively. Antiplasmodial evaluation of the two extracts revealed that the two extracts exhibited dose-dependent in-vivo suppressive, curative and prophylactive properties on the development of parasitaemia in Albino mice using a chloroquine sensitive strain of Plasmodium berghei (NK-65). TLC profile fingerprints of the aqueous extract revealed three distinct spots with Rf values of 0.23, 0.39 and 0.75 whereas that the chloroform extract revealed three distinct spots with Rf values of 0.33, 0.42 and 0.55 when it was developed in ethyl acetate: ethanol: water: ammonia (65:25:9:1).

Conclusion: These results represented the first conducted evaluation of the perception of TMPs of North-Western Nigeria on the causes, diagnosis and treatment of malaria, antiplasmodial and thin layer chromatographic profile fingerprinting studies on Plumeria rubra bark found in North-Western Nigeria. The findings are therefore expected to provide the necessary scientific basis for rational use of traditional health knowledge and use of medicinal plant resources of North-Western Nigeria in the treatment of malaria.

Author(s) Details

Umar Adam Katsayal
Department of Pharmacognosy and Drug Development, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria.

Mujitaba Suleiman Abubakar
Department of Pharmacognosy and Drug Development, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria.

Abubakar Ahmed
Department of Pharmacognosy and Drug Development, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria.

Ezzeddeen Mukhtar Abdurahman
Department of Pharmacognosy and Drug Development, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria.

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Drug Delivery Approaches | Chapter 01 | Modern Advances in Pharmaceutical Research Vol. 2

This review discusses the different biological barriers that affect the delivery of therapeutic agents through membranes, such as intestinal mucosa, Brain Blood Barrier (BBB), and mediators of transport such as efflux transporters and the approaches for overcoming such barriers. The approaches illustrated in this review include: utilizing natural occurring transporters to deliver drugs specifically to their targets, nucleoside analogues delivery, CYP-activated prodrugs that target drugs to the liver, modification of passive diffusion by efflux pumps, intestinal transporters such as PEPT1 and GLUT1, Carrier Mediated Transport (CMT) systems for transporting nutrients, vitamins or hormones into the central nervous system, tissue selective drug delivery, administration of an exogenous enzyme to reach the tumor site which is followed by systemic administration of non-toxic prodrugs (ADEPT, GDEPT and VDEPT), enzymes involve in the bioconversion of ester-based prodrugs for activation (hydrolysis) of prodrugs to their active forms, brain targeted Chemical Delivery Systems (CDS), amino acid prodrugs to improve oral bioavailability, sustained drug delivery and intravenous drug delivery. Furthermore, Receptor-Mediated Transcytosis (RMT) for efficacious delivery of Nano particles via the intestinal mucosa and BBB, and the prodrug chemical approach based on intra molecularity to deliver anti-cancer drugs is discussed.

Author(s) Details

Professor Rafik Karaman
Department of Bioorganic Chemistry, Faculty of Pharmacy, Al-Quds University, P.O. Box 20002, Jerusalem, Palestine and Department of Sciences, University of Basilicata, Viadell’Ateneo Lucano 10, 85100, Potenza, Italy.

Wajd Amly
Department of Bioorganic Chemistry, Faculty of Pharmacy, Al-Quds University, P.O. Box 20002, Jerusalem, Palestine.

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Computational Analysis of Evolutionary Relationship of a Family of Cold Shock Proteins in Ten Mammalian Species | Chapter 11 | Advances and Trends in Biotechnology and Genetics Vol. 2

Aims: This study was carried out to evaluate the evolutionary relationship of a family of cold shock proteins (CSP) in ten mammalian species using bioinformatics tools and softwares such as Genbank, FASTA, BLAST and MEGA 5.

Sample: Twenty protein sequences of both RBM3 and CIRP proteins of some selected mammalian species were downloaded from NCBI database.

Study Design: Computational analysis to evaluate the evolutionary relationship of the CSP was carried out by estimating the phylogenic relationship of CSP in the different mammalian species studied.

Place and Duration of Study: This study was carried out at the Department of Genetics and Biotechnology, Calabar.

Methodology: The molecular evolution and genetic analysis, version 5 (MEGA 5) software was used to determine the evolutionary relationship of both CIRP and RBM3 in the ten mammalian species studied by constructing phylogenic tree using the amino acid sequences of protein retrieved from NCBI.

Results: The highest identity (100%) was observed between Ovis aries and Bos Taurus; Rattus norvegicus and Mus-musculus while the least percentage identity was observed between Pan troglodytes and Bos taurus (84%). The phylogenic relationship using UPGMA based on Jones-Taylor-Thornton (JTT) matrix model revealed high relationship.

Conclusion: It was observed that evolutionary relationship of CIRP and RBM3 revealed high relatedness among the mammalian species studied.

Author(s) Details

E. A. Okon
Department of Biological Science, Cross River University of Technology, Calabar, Nigeria.

E. V. Ikpeme
Department of Genetics and Biotechnology, University of Calabar, Calabar, Nigeria.

O. U. Udensi
Department of Genetics and Biotechnology, University of Calabar, Calabar, Nigeria.

E. E. Ekerette
Department of Genetics and Biotechnology, University of Calabar, Calabar, Nigeria.

H. E. Etta
Department of Biological Science, Cross River University of Technology, Calabar, Nigeria.

E. P. Willie
Department of Genetics and Biotechnology, University of Calabar, Calabar, Nigeria.

M. Ozoje
Department of Genetics and Biotechnology, University of Calabar, Calabar, Nigeria.

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Diabetes Mellitus: Can Stem Cells be the Answer? | Chapter 10 | Advances and Trends in Biotechnology and Genetics Vol. 2

This review aims to enlighten the readers regarding the past, present and future of stem cells in the treatment of Diabetes. Diabetes is one of the leading causes of morbidity and mortality, affecting more than 415 million people worldwide.  It is estimated that one in ten adults will have diabetes by 2030. Diabetes is mainly due to reduction in β-cell mass which are responsible for insulin production. Exogenous administration of insulin is having good impact on restoring glucose homeostasis, but it does not entirely control the minute-to-minute fluctuations in systemic blood glucose. Recently cellular-based therapies have been established for exogenous insulin administration by modern pump technology. One of the most interesting therapies involves substitution of insulin producing islet cells by transplantation. But lack of donor material and lifelong immunosuppression made the technique unfeasible. These restrictions have led to exploration of other sources of β-cells, one of the prospects being the stem cells. Several types of stem cells have been used to make pancreatic β-cells, including human embryonic stem cells / induced pluripotent stem cells, pancreatic stem / progenitor cells, and non-pancreatic stem cells. There is also evidence of adult β-cells regeneration through β-cell replication and cellular reprogramming. Functional restoration of existing β-cells, transplantation of stem cells or stem cell-derived β-like cells might provide new opportunities for treatment. In conclusion it can be said that the research is still wide open to arrive at the efficient reprogramming of various types of stem cells to destine them towards functional β-cells.

Author(s) Details

M. Senthilnathan
Department of Veterinary Pharmacology and Toxicology, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India.

A. Ramadevi
Department of Animal Nutrition, Kerala Veterinary and Animal Sciences University (KVASU), Mannuthy, Kerala, India.

K. Srinivas
Department of Veterinary Public Health and Epidemiology, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India.

A. Thangamani
Department of Veterinary Gynecology and Obstetrics, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India.

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RNA-seq Evaluating Several Custom Microarrays Background Correction and Gene Expression Data Normalization Systems | Chapter 09 | Advances and Trends in Biotechnology and Genetics Vol. 2

Microarray gene expression technologies represents a widely used tool in transcriptomics and genomics studies worldwide. This technology is stable with the purpose of gene expression differential analysis because of their well-established biostatistics and bioinformatics analysis schemes. However, microarray reliability with regard that analysis typology, depend on probe specificity as well as applied data normalisation and/or background correction procedures. Then, we assessed the performance of 20 different microarrays background correction / gene expression data normalisation combination procedures from “linear models for microarray and RNA-Seq data analysis” package (limma), by comparing significantly differentially expressed genes detected by several custom microarray design strategies, depending on microarray probe size as well as probe set number per transcript model by assuming RNA-Seq approach as benchmark. Basing exclusively on a multivariate statistical clustering surveys, in R programing environment, we showed the pre-eminence of data normalisation (DN) as opposed to noise background correction/subtraction (BS) in microarray expression analysis. Although the combination between (i) gene expression data normalization and (ii) background subtraction procedures (BS+DN), improves the agreement between heterogenic microarray platforms as well as RNA-Seq platform in calling significantly modulated genes, quantile normalisation system combined with all processed background correction procedures has been discriminated as exhibiting highest sensitivity with RNA-Seq (p < 0.05). In conclusion we showed the pre-eminence of microarray data pre-processing step in gene expression differential analysis by according a priority to data normalisation procedure especially to quantile normalisation system contributing in stabilizing gene expression differential analysis results with regard heterogenic custom microarray design strategies (heterogenic microarray platforms).

Author(s) Details

Noel Dougba Dago, Msc., Ph.D
Unité de Formation et de Recherche (UFR) des Sciences Biologiques, Département de Biochimie-Génétique, Université Peleforo Gon Coulibaly BP1328 Korhogo, Côte d’Ivoire.
Laboratory of Functional Genomic, Department of Biotechnology, University of Verona, Strada Le Grazie 15 CàVignal 1, 37134, Verona, Italy.

Dr. Martial Didier Yao Saraka
Unité de Formation et de Recherche (UFR) des Sciences Biologiques, Département de Biochimie-Génétique, Université Peleforo Gon Coulibaly BP1328 Korhogo, Côte d’Ivoire.

Dr. Nafan Diarrassouba
Unité de Formation et de Recherche (UFR) des Sciences Biologiques, Département de Biochimie-Génétique, Université Peleforo Gon Coulibaly BP1328 Korhogo, Côte d’Ivoire.

Antonio Mori
Department of Neurological, Biomedical and Movement Sciences, University of Verona, Strada Le Grazie 8, 37134, Verona, Italy.

Dr. Hermann-Désiré Lallié
Unité de Formation et de Recherche (UFR) des Sciences Biologiques, Département de Biochimie-Génétique, Université Peleforo Gon Coulibaly BP1328 Korhogo, Côte d’Ivoire.

Prof. Professor Lamine Baba-Moussa
Laboratoire de Biologie et de Typage Moléculaire en Microbiologie, Faculté des Sciences et Techniques, Université d’Abomey-Calavi, Cotonou, Benin.

Dr. Massimo Delledonne
Laboratory of Functional Genomic, Department of Biotechnology, University of Verona, Strada Le Grazie 15 CàVignal 1, 37134, Verona, Italy.

Giovanni Malerba
Department of Neurological, Biomedical and Movement Sciences, University of Verona, Strada Le Grazie 8, 37134, Verona, Italy.

Edouard Kouamé N’Goran
Unité de Formation et de Recherche (UFR) des Sciences Biologiques, Département de Biochimie-Génétique, Université Peleforo Gon Coulibaly BP1328 Korhogo, Côte d’Ivoire.

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Solid State Fermentation Based Olive Pomace Using Streptomyces Strains: A Preliminary Study | Chapter 08 | Advances and Trends in Biotechnology and Genetics Vol. 2

This study represents a preliminary investigation aimed to assess the possibility to recycle and valorize olive pomace by solid state fermentation (SSF) using Streptomyces strains. The olive pomace was collected from an olive pressing factory (super press system) during the olive fruit pressing season. The study was carried out at IMBE, University Aix Marseille-France, between April 2013 and June 2013 and at LMA, University of Bejaia-Algeria in September 2013. Three Streptomyces strains designated S1M3I, S1M3II and S1M3III were cultured on solid state fermentation based olive pomace at 30°C for 10 days, and subsequently, the lignocellulolytic enzyme activities (xylanase, CMCase and laccase), the viability of the microorganisms and the pH of the resulting substrates, were determined. The fermented substrate pH values remained significantly stable (p ˂ 0.05) throughout the fermentation period for the three strains; they were fluctuated between 6.54 and 6.99. The viability of all Streptomyces strains studied, decreased significantly (p ˂ 0.05) during the first four days of incubation, to reach up 0 cfu/mL of viability and 0 U/g enzymatic activities (xylanase, CMCase and laccase activities) were recorded for the three strains. Streptomyces strains, under the experimental conditions (30°C, pH 7 and 75% of moisture), were unable to grow and produce lignocellulolytic enzymes in solid state fermentation based olive pomace due to the mycelial morphology and Streptomyces developmental cycle, no neglect, the environmental factors. These preliminary results suggest that SSF – Streptomyces system is not suitable for conversion of solid waste from olive processing industry and to produce lignocellulolytic enzymes.

Author(s) Details

Lamia Medouni-Haroune
Laboratoire de Microbiologie Appliquée, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, 06000 Bejaia, Algérie.

Farid Zaidi
Département des Sciences Alimentaires, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, 06000 Bejaia, Algérie.

Sevastianos Roussos
Equipe Eco Technologies et Bioremédiation, Faculté St Jérome, Campus Etoile, Aix Marseille Université & Université Avignon, IMBE UMR CNRS-7263/IRD-237, Case 421, 13397 Marseille Cedex 20, France.

Véronique Desseaux
Institut des Sciences Moléculaires de Marseille, Faculté des Sciences et Techniques, St Jérome, Biosciences UMR CNRS 6263. Université Paul Cézanne, 13397 Marseille Cedex 20, France.

Sonia Medouni-Adrar
Laboratoire de Biomathématiques, Biophysique, Biochimie, et Scientométrie (L3BS), Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, 06000 Bejaia, Algérie.

Mouloud Kecha
Laboratoire de Microbiologie Appliquée, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, 06000 Bejaia, Algérie.

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Kerosene: A Study of Tissue Histology and Serum Vitamin and Heavy Metal Levels of Female Wistar Rats Chronically Exposed | Chapter 07 | Advances and Trends in Biotechnology and Genetics Vol. 2

Background: Kerosene a commonly available product used for a variety of purposes in many parts of Asia and Africa is sometimes sold in beverage bottles and jerry cans in both commercial and residential places due to inadequate number of filling stations. Therefore excessive exposure through both dermal and oral routes is common.

Objective: This study was embarked upon to ascertain the impact of trace amount of kerosene on tissue histology as well as serum vitamin and heavy metal levels in female Wistar rats.

Methods: Kerosene (0.4 mL/kg body weight) was administered to rats either through the oral or dermal route daily for a period of 30 days. The effects of kerosene administration on tissue histology; serum vitamin levels and serum concentrations of heavy metals were evaluated using hematoxylin- eosin staining technique; high performance liquid chromatography (HPLC); and atomic absorption spectrometry (AAS) respectively. Student’s t test and analysis of variance were used for statistical analysis of data. P <0.05 was considered significant.

Results: Histopathologic presentation such as pulmonary congestion, severely stunted villi, congestion of coronary vessels, and diffuse spongiosis of the cerebral cortex were observed in kerosene administered groups while control group featured no visible lesion. Results of heavy metals revealed that although As, Al and Cd were not significantly different in kerosene exposed groups compared with control, Si was significantly lower (oral), and significantly higer (dermal) compared with control. In addition, using analysis of variance (ANOVA) all estimated vitamins (except pantothenic acid) were significantly different in kerosene exposed groups compared with control.  

Conclusion: The results of this study indicate that exposure to this product either through the oral or dermal route may be detrimental to health as it induced adverse alteration in vitamin and heavy metal levels as well as distorted tissue histo-architecture.

Author(s) Details

Ayobola A. Iyanda
Department of Chemical Pathology, College of Health Sciences, Ladoke Akintola University of Technology, Osogbo, Nigeria.

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Assessment of Methaemoglobin and Carboxyhaemoglobin Levels among Pregnant Women Infected with Hepatitis B Virus | Chapter 06 | Advances and Trends in Biotechnology and Genetics Vol. 2

Aims: The goal of this research is to determine plasma levels of MetHb and COHb in pregnant women with hepatitis B, which might enhance oxidative stress and hypoxemic condition of this state if it is not ameliorated on time.

Study Design: Prospective case-control study.

Place and Duration of Study: Antenatal clinic at Primary Health Centres, Sagamu, Ogun State, Nigeria between February, 2015 and August, 2015.

Methodology: Blood levels of MetHb, COHb and bilirubin were determined in ninety four (94) participants (aged 18-40 years), divided into three groups: 33 pregnant women infected with hepatitis B virus, 30 apparently healthy pregnant women and 31 age matched non pregnant women apparently healthy controls. Blood levels of MetHb, COHb and bilirubin were determined using standard spectrophotometric method.

Results: There was progressive increase and decrease in mean blood levels of (TBil and MetHb) and mean blood levels of COHb respectively from controls through pregnant subjects with HBV. PCV and DBil had no specific pattern of differences across the groups.

Conclusion: This study showed a slight increase in blood levels of MetHb in pregnant women with hepatitis B and apparently healthy pregnant women compared to non-pregnant controls, which might enhance oxidative stress and hypoxemic condition of this state. It would also be helpful to incorporate MetHb screening as routine tests for better management of pregnant women especially with HBV.

Author(s) Details

Adedeji David Atere
Department of Medical Laboratory Science, Achievers University, Owo, Ondo State, Nigeria.

Franklin Kayode Ayenogun
Department of Medical Laboratory Science, Achievers University, Owo, Ondo State, Nigeria.

Bolaji David Akinbo
Department of Medical Laboratory Science, Achievers University, Owo, Ondo State, Nigeria.

Adaobi Mary-Joy Okafor
Department of Biological Sciences, Convenant University, Otta, Ogun State, Nigeria.

Kelvin Ifeanyichukwu Egbuchulem
Department of Surgery, University College Hospital, Ibadan, Oyo State, Nigeria.

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