Mutations in the TP53 gene had been attributed to the development of liver cancer. Hepatocellular carcinoma (HCC) and liver cirrhosis are liver diseases having high mortality rates in several populations. There is no information on the TP53 gene polymorphism among liver diseases patients in Calabar, Nigeria. This study investigated the genetic polymorphism of TP53 among HCC and liver cirrhosis in Calabar. This research was carried out in the University of Calabar Teaching Hospital, Calabar. Blood was collected from 35 clinically diagnosed hepatocellular carcinoma and 10 cirrhosis patients and 10 healthy controls. DNA was extracted from all blood samples. Polymerase Chain Reaction (PCR) was performed. The amplicon was digested using Hae III restriction enzyme and the genotypic and allelic frequencies were determined. In liver diseases patients, male was 68.9% (31), female (31.1%; 14), sex ratio (2.2: 0.5), mean age was 41.51 ± 2.13 years and odd ratio of 1.25. Marital status of patients were 33(73.3%), 10(22.2%), and 2(4.4%) for married, single, and widowed respectively. Ethnicity of patients were Yakurr, Efik, Boki, Ogoja, Annang, Ibibio and Igbo having 22.2%(10), 20%(9), 13.3%(6), 13.3%(6), 8.8% (4), 2.2%(1) and 2.2%(1) respectively. Approximately, 64.7% (30) of the chronic liver diseases were from the Central and Northern part of Cross River State. The risk factors were HCV infection, HBsAg+, alcoholism, smoking, consumption of groundnuts that may have been contaminated with aflatoxin and family history of the disease. PCR product yielded 254bp and digested PCR product showed homozygous TT mutation (27), heterozygous GT mutation (17) and homozygous GG wild type (1) in cases. The overall TP53 gene mutation frequency was 46.32% (44). The frequency of G allele, T allele, GG, GT and TT genotypes were 0.21, 0.79, 0.04, 0.33 and 0.62 respectively among cases, while GG (wild type) was only detected among controls in the study population. The genotypic and allelic frequencies conform to Hardy-Weinberg equilibrium meaning that the forces of evolution were not acting on the locus. There were significant differences in the genotypic proportions of the TP53 gene polymorphism among patients and controls. This study on the TP53 gene polymorphism will serve as baseline information on the molecular etiology of hepatocellular carcinoma and liver cirrhosis in Cross River State.

The present study investigated the rate of change (mutation) in TP53 and the associated functional partner genes and revealed that they play very significant role in hepatocellular carcinoma and liver cancer disease conditions in humans. Genetic correlation analysis has shown that there is very high association, strong relationship with significant impact between TP53 and the functional partner genes. The strength of association between TP53 gene and other functional partners gene was found to be high (> 0.5) in normal liver but low than in cancerous liver. The proteomic parameters of TP53 and other functional partner genes such as the molecular weights, number of amino acids, theoretical pl, total number of atoms, total number of positive and negative amino acids residues, extinction coefficients, estimated half-life, instability index, aliphatic index and hydropathicity were revealed in the study and viewed using the circos visualizer which showed bigger strands for genes with high molecular weights. The expasy.org prosites analysis of the TP53 and associated functional partner genes revealed the functional domain protein site reaction hotspots with the corresponding amino acids. The main prosites reaction hotspots were the protein kinase II phosphorylation site (PKC) which was similar for all the associated genes. Other domain reaction hotspots for TP53 and associated genes includes N-glycosylation sites, N-myristylation sites, N-Amidation sites, Tyrosine II kinase phosphorylation sites I and II, the casein II protein kinase phosphorylation sites (CK2) and the cAMP and cGMP phosphorylation sites. Two principal components were used to explain the variations in dimensionality of the TP53 and associated genes. ATM gene showed the highest loading value in PC1 while EP300 gene exerts the highest impact in terms of dimensionality in PC2. The principal component axes delineated the genes into two major cluster sets. Major cluster one had four genes which major two had 6 genes. The intensity of interactions among the genes to induce hepatocellular carcinoma and liver cancer was demonstrated using interactive heatmap with red colour depicting intense interactions, black colour depicting moderate interactions and green colour depicting slow interactions.

A survey was carried out to investigate the knowledge, perception and interest of secondary school students about biotechnology. A total of 334 questionnaires were distributed to students in Senior Secondary 3 classes from eight secondary schools within Calabar metropolis. Data was collected and analyzed using SPSS version 7.5. Results revealed 105 (34.21%) of students had limited knowledge of medical biotechnology, genetic engineering and genetically modified products. 91 (30.03%) of the students agreed that biotechnology is the use of living organisms to produce goods and services while 102 (33.41%) accepted that biotechnology is a new technology based on biology 62 (20.39%) were willing to embrace the applications of biotechnology. 152 (50.03%) do not agree that biotech will improve services for mankind; 90 (29.51) had no idea about the applications of biotechnology; 31 (10.24%) students disagreed that cloning results in perfectly identical individuals while the highest positive responses; 92 (30.31%) was recorded for questions regarding in vitro fertilization. The students also showed a very low interest 76 (25%) in pursuing biotechnology as a degree in the University. Generally students' knowledge, perception, and interest in Biotechnology were low among Secondary School students in Calabar. There is a need to immediately increase enlightenment and emphasis on the applications of biotechnology among Secondary school students to enable them appreciate the benefits of biotechnology.

Retinoblastoma is the most common intraocular malignancy in infancy and childhood. The incidence varies in different populations. Mutations of the retinoblastoma susceptibility gene have been implicated in malignant retinoblastoma. This research seeks to identify any mutations that could be present in exon 19 and have implications in the pathogenesis of retinoblastoma among children in Calabar. Children (9) with retinoblastoma attending the University of Calabar teaching Hospital (UCTH), Dept of Opthalmology. 30 unrelated and unmatched controls with no history of retinoblastoma were recruited into the study. 23mls of blood was collected from each child, genomic DNA was extracted from blood, PCRs and sequencing were performed on exon 19. The nucleotide sequences of the RB1 gene were decoded from the chromatogram using Bioedit software and aligned manually. Multiple sequence alignment was performed using CLUSTAL W. The RB1 gene mutation frequencies were 2(22.2%) and 3(33.3%) for missense mutations and deletions respectively. Sequencing revealed two missense mutations namely: g.98A>G (p.Y33C) and g.154A>G (p.I52V) in a male patient and a female patient. Deletions include: g.1delC, g.1-12delCAGGAAAACCA, g.45-46delAA, g.1421delTTATTAAA and g.1-55delCAGGAA.....TTC were all observed in male (bilateral) cases of retinoblastoma 3(11.1%). These mutations were absent in 6(66.6%) children and the control subjects. Two missense and five deletions were observed in four males and a female patient. The RB1 gene mutation frequency was low among the retinoblastoma children, implying that these mutations were not directly responsible for retinoblastoma, the main causal mutation may be present in other exons. Exon 19 needs to be investigated in a larger population.

Data was collected on ABSTRACT Retinoblastoma is an eye disease causing blindness and death in children. The screening of the RB1 gene in retinoblastoma children, relatives and controls lead to the documentation of more than 900 mutations in different populations, which are important for genetic counselling, characterization of phenotypic-genotypic relationships and clinical management of the disease. There is no information on the RB1 gene polymorphism among retinoblastoma children in Calabar, Nigeria. This study investigated the genetic polymorphism of RB1 susceptibility gene and its involvement in the molecular pathogenesis of retinoblastoma in Calabar. This research was carried out in the University of Calabar Teaching Hospital (UCTH) Calabar, Nigeria. Blood was collected from 9 clinically diagnosed retinoblastoma children, 30 children and 23 adults as controls. DNA was extracted from all the blood samples, PCR was performed yielding a 485bp amplicon that was digested using the M1u CI restriction enzyme. The digested fragments were visualized in 1.5% agarose and the genotype and allele frequency was determined. The genotype frequencies among the retinoblastoma children were 0(0%), 3(33.33%) and 6(66.67%) and in the controls were 0(0%), 13(24.53%) and 40(75.47%) for GG, GA and AA respectively. The G allele was low (0.17and 0.12 in patients and controls). There were no significant differences in the genotypic proportions of the RB1 gene polymorphism among patients and controls. The RB1 gene polymorphism may not be directly involved in the development of retinoblastoma however this requires further investigation.

Background: Primary congenital glaucoma (PCG) is an inherited ocular congenital anomaly of the trabecular meshwork and anterior chamber angle, which results in optic nerve damage due to increase intraocular pressure if not properly managed. We explore CYPIBI gene mutations in three Original Research Article Duke et al.; JAMMR, 27(3): 1-10, 2018; Article no.JAMMR.35069 2 cases of primary congenital glaucoma, their family members in Calabar, helping to discuss future options for the disease management. Methods: Clinical and molecular screenings were conducted on three cases of PCG, six parent and six siblings of the PCG patients recruited for this study. 51 unrelated, age-matched controls comprising 30 children control and 21 adult control were selected from individuals attending the eye clinic for general eye examinations that has no glaucoma or history of glaucoma. 2-3 ml of blood was collected from each participant, DNA extracted from blood, and PCR amplifications were carried out on exon 3. 25 μl of amplicon was utilized for bidirectional sequencing. The nucleotide sequences of the CYPIBI gene were edited from chromatograms using Bioedit software. Multiple sequence alignment and pairwise comparison of CYPIBI gene was carried out in the MEGA 6 software. Statistical analyses were performed using SPSS version 20 software. Significant level was set at P<0.05. Results: Two missense mutations of CYPIBI gene namely: g.291G>C (p.Q97H) and 344C>T (P.T115M) were observed in this pilot study after the in-silico analysis. The g.291G>C mutation that resulted in an amino acid substitution of glutamine by histidine at position 97(p.Q97H) was observed in all the 3 cases of PCG, their parent (six) and siblings. The g.344C>T was detected in only one PCG patient. Two deletions of CYPIB1 gene namely: g.378-380delATG and g.535delG were also observed in this study. The g.378-380delATG was detected in all the PCG cases, their parents while g.535delG was seen in only one PCG patient that was 28 months old boy. These CYPIBI gene mutations were absents in all the controls. Conclusion: This pilot study identified some CYPIBI gene mutations, which were peculiar to PCG cases, their parents and other family members. It suggests hereditary form of the disease although further studies are needed. This forms a baseline research for further molecular studies among PCG patients.

This research involved modification of a standard protocol for DNA extraction in plants for its suitability as an extraction method for DNA in human whole blood in which some of the plasma was removed. Human blood samples were obtained from  a 100 apparently healthy individuals residing in Calabar. The modified DNA procedure yielded good quality genomic DNA which was used in carrying out allele specific polymerase chain reaction which also yielded good quality amplicons. This method is simple and suitable for the extraction of DNA from human red cell.

The association between Obesity and Blood Pressure is well documented and a series of Obesity indices are used as risk factors or indicators of Blood Pressure or the presence of Hypertension. The objective is to observe for any relationship between Blood Pressure levels and its association with various indices of obesity by gender among traders of Watt market, Calabar. A total of 300 subjects (150 men and 150 women), aged 20-60 years were selected for the study using a convenient and random sampling technique in watt market Calabar. Weight, Height, Waist Circumference (WC), Hip Circumference (HC) and Blood Pressure were measured for each participant. Four anthropometric variables, Body Mass Index (BMI), Waist to Hip ratio (WHR), Waist to Height ratio (WHtR) were calculated and used to evaluate their association with Blood Pressure (BP). Pearson Correlation and Paired t-test analysis were used; Correlation was calculated between Systolic and Diastolic Blood Pressure and various indices of obesity. All four obesity indices used for this study were positively correlated with Blood Pressure and Hypertension. Waist to Height ratio in women and BMI in men had the strongest correlations or associations with BP. The association of Waist to height ratio in women and Body Mass Index (BMI) in men remained significant after adjustment for each of the other variables. However, BMI had the strongest correlation with continuous Blood Pressure in both genders. The association of WHR, and WC with BP and the prevalence of hypertension were a little weaker than those of BMI and WHtR in both men and women respectively. Blood pressure levels and Obesity indices increased steadily with age both in males and females. Variation in SBP was significantly correlated with all the obesity indices used in the study (BMI, WC, WHR, WHtR) in both males and females except WHtR in females. Also variation in DBP was significantly correlated with all indices of obesity (BMI, WC, WHR, WHtR) in males, but in females it had strong correlation with BMI, WHR, and WHtR. Percentage prevalence of obesity increased with age more in females than in males.

Background: Tuberculosis (TB), though a curable infectious disease, remains one of the leading causes of death in adults. It is also a major public health concern in Nigeria. This study was aimed to determine the prevalence rate of tuberculosis among patients attending the out-patient Department of Dr. Lawrence Henshaw Memorial Hospital Calabar. Materials and Methods: A retrospective study was carried out consisting of all documented cases of tuberculosis from January 2005 to April 2015. Results: Out of 20185 patients tested, a total of 5,004 cases of tuberculosis was recorded within the period of this study with a prevalence rate of 24.8%. The prevalence for each year during this study were as follows: 2005 (37.5%), 2006 (30.9%), 2007 (26.2%), 2008 (23.1%), 2009 (23.0%), 2010 (20.5%), 2011 (16.6%), 2012 (20.5%), 2013 (22.9%), 2014 (21.8%) and 2015 (44.6%). HIV co-infection was more prevalence in males than females, while majority of the disease site was pulmonary tuberculosis (PTB). The highest mortality was recorded in 2012 (18.2%). Within the ethnic groups assessed, the prevalence was higher among the Efiks. Conclusion: The prevalence of TB/ HIV co-infection and subsequent mortality within the years under study is traumatizing and thus requires prompt measures in combating the situation.

Introduction: Congenital cataract is particularly important as it is the leading cause of blindness and severe visual impairment in children in Africa, accounting for at least 35% of blindness and severe visual impairment. This study seeks to identify mutations in the gamma crystallin gene that are associated with congenital cataract in some children attending the Eye Clinic in Calabar. Methods: Children (11) with congenital cataract attending the University of Calabar teaching Hospital (UCTH) Pediatric Ophthalmology and Strabismus Unit, Department of Opthalmology. 11 unrelated and unmatched controls that had no history of cataract were recruited into the study. 2-3 ml of blood was collected from each child, DNA was extracted from the blood, and PCR amplifications were carried out. About 25 μl of the PCR amplicon was used for sequencing. Multiple sequence alignment and pairwise comparison of the crystallin gene was carried out in the Mega 7 software. Results: The PCR amplication revealed a 742 bp amplicon which was sent for sequencing. In silico analysis revealed a substitution of guanine by cytosine at position 248 (g.248 G>C) that resulted in an amino acid substitution of arginine by proline at position 83 (p.R83P) in 7 (63.63%) out of the 11 patients. This substitution was absent in 4 patients (36.4%) and in the control subjects. Conclusion: This study identified a G>C substitution at position 248 in the crystallin gene in 7 out of 11 congenital patients and this forms a baseline research for further molecular studies among cataract patients in Calabar.

Background: Breast cancer type 1 (BRCA1) gene also known as breast cancer type 1 susceptibility protein homologue plays important role in DNA double-strand break repair, homologous recombination, chromatin remodeling, cell cycle regulation and transcription regulation.Objective: Considering the importance of this protein, the present study was undertaken to analyze BRCA1 gene of different mammalian species by assessing the identity and similarity, phylogenetic relationship, physicochemical properties, predict the motifs, secondary and the tertiary structures.Materials and Methods: Seventeen nucleotide and protein sequences of BRCA1 gene of different mammalian species were retrieved from National Centre for Biotechnology Information (NCBI).Multiple sequence alignment was done using CLUSTAL W software, while identity and similarity was determined by constructing a pair wise comparison.Results: Results obtained at the end of the experiment showed that there was no percentage identity or similarity that was less than 70%.The phylogenetic relationship of BRCA1 gene of the mammalian species clustered into aquatic, herbivores, carnivores and omnivores, respectively.The highest time of divergence (95MYA) of the BRCA1 gene was observed between the BRCA1 gene of killer whale and human, while the least time of divergence (4.6MYA) was observed between the BRCA1 gene of cattle and American buffalo.Physicochemical properties of BRCA1 proteins in the five mammalian species (cattle, sheep, pig, American buffalo and human) were found to be unstable, hydrophilic and intracellular in nature.The following motifs were present at various positions in the BRCA1 gene of the five mammalian species; zinc finger RING, BRCT domain, N-myristoylation site, N-glycosylation, cAMP and cGMP dependent protein kinase phosphorylation site, tyrosine kinase phosphorylation, casein kinase 2 phosphorylation site, protein kinase C phosphorylation site and cell attachment sequence but leucine zipper pattern and microbodies c-terminal motifs were found in only human BRCA1 gene.The BRCA1 secondary structure contained the alpha helix, extended strand and random coil.Conclusion: Based on the results obtained, it can be deduced that BRCA1 gene has identical homologue, functional similarity and highly conserved in these mammalian species.

The utilization of molecular markers-assisted genetic diversity study is critical and imperative for a more robust selection and breeding programmes in aquaculture.Twenty C. gariepinus species were randomly selected, 5 each from four Rivers/locations-Okurikan (OKU), Lemna (LMN), UNICAL Fish Farm (UFF) and MCC cultured fish site, in Cross River State, Nigeria for genetic diversity study using RAPD technique.Genomic DNA extraction and purification from caudal fin tissue sample was performed using ZR Genomic DNA tissue extraction Kit, primed using OPA09, OPA11 and OPA13 RAPD primers and amplified using PCR.The amplified fragments were ran on gel electrophoresis and visualized under ethidium bromide dye.Thirty four bands were generated by the three RAPD primers from the four C. gariepinus populations of which 28 were polymorphic, accounting for 82.35%.Polymorphism revealed by each fish population ranged from 92.86-100% while, gene diversity estimates ranged from 0.246±0.16-0.385±0.09.Intra-population similarity index ranged from 42.49-78.4%while, the inter-population similarity index across all population combinations ranged from 52.32-71.69%.These result also revealed that as genetic distances increases, genetic identities narrows between the wild and the cultured species.UPGMA dendrogram generated two major clusters with C. gariepinus species from OKU, UFF and MCC in cluster-1 and C. gariepinus species from LMN in cluster-2.Implicitly though, our results suggest that there were several genetic diversity in the populations studied, especially between the cultured and the wild, which could encourage selection of C. gariepinus from OKU and LMN populations for the purpose of breeding and improvement.

Morphological and behavioral genetic traits of simple inheritance indicate ethnic variation and have been widely employed in population variation studies.Five selected genetic traits of simple inheritance, ABO and Rh (D) blood phenotype was investigated to establish the inheritance pattern, prevalence and to assess association between the traits, ABO rhesus phenotypes and gender of 45 unrelated families living in Calabar.Blood group O was the most prevalent (55.2%) followed by B (21.6%),A (18.8%) while the least was AB (4.4%).The majorities (91.6%) were Rh (D) positive and 8.4% were Rh (D) negative.The frequency of the 5 genetic traits were earlobe (69.2% free earlobe, 30.8% attached earlobe), hand-clasping (51.6% right hand-claspers, 48.4% left hand-claspers), dimples (21.2% had facial dimples, 78.8% had no dimples), mid-digital hair (presence in 92%, absence in 8%) and tongue rolling (48.4% rollers, 51.6% non-roller).There was association between earlobe and blood group, sex and hand clasping, sex and facial dimples and between tongue rolling and sex.Chi-square analysis of the inheritance pattern of these genetic traits provides strong evidence for a familial and probably a genetic component in the control of these traits.The pattern also showed that the traits are inherited in dominance versus recessive manner.

Hypertension results from an interaction of many risk genes and environmental factors. The M235T allele of the angiotensinogen gene is thought to increase plasma levels of the angiotensinogen which is associated with hypertension. The angiotensinogen is an important substrate for renin in the RAAS that is finally converted into angiotensin II that plays a key role in the control of blood pressure. This study was designed to measure plasma angiotensinogen levels in an adult population in Calabar and Uyo (South-South), Nigeria in relation to the M235T allele and hypertension. Out of a large population of 1224 participants who had been genotyped for the M235T polymorphism, plasma was collected from a sample of 300 consisting of 150 patients and 150 controls. Protein A sandwich enzyme linked immunosorbent assay was carried out with the plasma samples to measure the angiotensinogen levels. Age, BMI, M235T allele, blood pressure and O.D values were compared between controls and patients using the independent t test. The absorbance values of plasma angiotensinogen were significantly higher in the patients (0.71) with M235T allele than in the controls (0.53). Further research still needs to be carried out to determine the actual concentration of the protein in the participants.

Background: Hypertension is a public health challenge due to its high prevalence, and is a major risk factor for cardiovascular diseases. This study was designed to determine the frequency of the I/D polymorphism of the angiotensin-converting enzyme gene and its association with hypertension in a sample population of Calabar and Uyo, South-South Nigeria. Materials and Methods: A population-based case control design consisting of total of 1224 participants, 612 each of patients and controls, were randomly recruited from hypertension clinics and the general population. The I/D polymorphism was investigated using polymerase chain reaction. Multiple regression and odds ratio (OR) was applied to test whether the ID genotypes were predictors of hypertension. Results: The I/D genotype frequencies were 73(12%), 262(43%) and 277(45%); 74(12%), 303(50%) and 235(38%) for the II, ID, DD genotype in patient and control groups, respectively. A higher frequency of the ID genotype was observed in controls of which 208(61%) were females. By multiple regression analysis, age was a predictor for SBP in patients, r = 0.596, and DBP in controls, r = 0.555. Gender, Body mass index, I/D genotypes were not significant predictors for hypertension but the I/D polymorpism was associated with an increased risk for hypertension with an OR of 1.15 95%CI (0.924-1.456).Conclusion: The I/D polymorphism of the angiotensin-converting enzyme gene was a risk factor for hypertension in the sample population of Calabar and Uyo. This research will form baseline information for subsequent molecular studies in this population.

A common molecular variant of the angiotensinogen gene had been reported to predispose some ethnic groups to hypertension. This case–control study was designed to determine the frequency and association of the angiotensinogen M235T allele with hypertension in residents of Calabar and Uyo cities, south–south Nigeria.

OBJECTIVES:The angiotensin II protein is a vasoconstrictor that exerts most of its influence through the angiotensin II type 1 receptor (AT1R). Inconsistent association between the A1166C polymorphism of the AT1R gene and hypertension has been reported among various populations but not among the peoples of Calabar and Uyo. This study was designed to determine the frequency of the A1166C polymorphism of the AT1R gene and its association with hypertension in a sample population of Calabar and Uyo.MATERIALS AND METHODS:A population-based case control design consisting of total of 1224 participants, 612 each of patients and controls were randomly recruited from hypertension clinics and the general population. Genotyping of the A1166C allele of the AT1R gene to identify variants was performed using polymerase chain reaction and restriction enzyme digestion. Multiple regressions were applied to test whether the A1166 genotypes were predictors of hypertension.RESULTS:99% of the study population had the wild type AA genotype, and 1% was AC heterozygous carriers of the A1166C polymorphism.CONCLUSION:The A1166C polymorphism was not a predictor of hypertension in the sample population of Calabar and Uyo.

HIV/AIDS, a disease first mentioned in 1981 has remained the most stigmatized illness especially in sub Saharan Africa. To assess the knowledge and effect of the large pool of information on HIV/AIDS in the lives of teenagers. A total of 1748 male and female students aged 15 to19 years in 12 secondary schools in Port Harcourt and Calabar randomly selected were used for this study between May and December 2009. A self administered questionnaire comprising 18 multiple choice questions was developed for the study. Among the 1748 participants general knowledge on HIV/AIDS was high (79.9%). Less than 15% acknowledge its presence in semen and vaginal secretions though a greater than 74% believe HIV/AIDS is transmitted through sex. Condom use is low (15.7% females and 33.3% males). More girls (56.4%) prefer to abstain from sex until marriage than boys (34.9%). Only 38% agreed to voluntary HIV testing. Only 37.8% females and 26.2% males would support and care for friends and relations who are HIV-positive, would not spread the infection and would seek medical advise for themselves if infected. Information on HIV/AIDS should be encouraged. The youths should be targeted. This will reduce the prevalence. 20-22 Key words: HIV/AIDS, gender differences, students, Niger Delta, Nigeria.

This study was aimed at qualitative evaluation of the ethanol seed, leaf and pulp extracts of C. papaya for bioactive compounds and also to investigate their effect on the haematology in male albino rats. A 3 x 4 factorial experimental layout using randomized complete design was adopted. Results show that the phytochemicals found in seed, leaf and pulp were almost the same but however, in varying proportions. Present result also revealed that there were significant effects (p < 0.05) of the extracts on the heamatology of the treated rats, which was blamed on the varying and different variants ofbioactive compounds found in the extracts they were administered with. Suggestively, C. papaya extracts could be used to enhance the production of selected blood parameters, taking issue of dosage into consideration.