Vol.5 No.1 – 12 : Gelatin-coated magnetic nanoparticles-based DNA isolation method: A comparison with commercial DNA isolation kits from whole blood

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By: Kelly S Levano1,2, Luis Jaramillo-Valverde1,2, Elizabeth Delgado1,2, Heinner Guio1,2*

1INBIOMEDIC Research and Technological Center, Lima, Peru;

 2ALBIOTEC, Lima, Peru;

Abstract

DNA isolation is the first step for most molecular tests, thus indicating the importance of using a reliable and suitable method for this procedure. The use of magnetic nanoparticles for blood DNA isolation offers several advantages including: subjecting little mechanical stress to your sample, obtaining higher quality and quantity of DNA and non-laborious procedures. In this study, we isolate genomic DNA using gelatin-coated magnetic nanoparticles from blood samples and test its use compare with three commercial blood DNA isolation kits.  DNA purity and yield were assessed by measuring absorbance at A260/A280 and by agarose gel electrophoresis. The suitability of the isolated DNA for downstream applications was analyzed by end-point PCR and Sanger sequencing. In this study, gelatin-coated magnetite particles for genomic DNA isolation provide an efficient, simple, and inexpensive method that does not require the use of commercial blood DNA Isolation Kit.

Gelatin-coated magnetic nanoparticles-based DNA isolation method A comparison with commercial DNA isolation kits from whole blood

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Vol.5 No.1 – 11 : Assessment of cell-free DNA for early detection of ovarian cancer in women

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By: Waleed M Fathy,1 Sabah Farouk,2 Gamalat Elgedawy,3 Ayman Abd-Elhakim Abd-Elghany, 4 Amira Ahmed2

1Clinical Pathology Department, Faculty of Medicine, Menufia University, Egypt

2Molecular Biology Department, Genetic Engineering and Biotechnology Research Institute, Sadat City University Egypt

3Biochemistry Department, National Liver Institute, Menufia University, Egypt

4Obstetrics and Gynecology Department, General Organization of Teaching Hospital, Egypt

Abstract

Objectives: To assess circulating cell-free DNA’s diagnostic potential in Egyptian women with ovarian cancer. Background: Ovarian cancer (OC), one of the most common cancers worldwide, is the most lethal form of gynecological cancer, but the early detection of ovarian cancer would significantly decrease its mortality rate. Circulating plasma cell-free DNA (cfDNA) is nucleic acids in peripheral blood that originate from cell death caused by injury, apoptosis, and necrosis. Circulating cfDNA is normally found in small amounts in the blood of healthy individuals, although increased cfDNA levels have been reported in patients with various clinical conditions, including infection, inflammation, malignancy, connective tissue diseases, ischemic stroke, myocardial infarction, pregnancy-associated disorders, and hemodialysis. Subjects and Methods: This study was conducted on 50 patients with OC, 25 patients with benign ovary disease (BOD), and 25 apparently healthy women used as a control group. All participants were tested for AFP, HCG, CA125, LDH, and circulating cfDNA, which were measured using real-time quantitative Polymerase chain reaction (RT-qPCR). Results: Circulation cfDNA rises more dramatically between OC cases than in both BOD cases and healthy controls, and the results indicate that circulating cfDNA was significantly higher in the OC group (p < 0.001) than both the BOD and control groups. The receiver operator of characteristics (ROC) curve analysis of circulating cfDNA revealed that, at a cut-off value of > 4.13 (fold expression), the sensitivity and specificity for differentiation of OC cases from non-cancer subjects were 97.3% and 92%, respectively. A significant positive correlation was found between circulating cfDNA and CA 125.Conclusion: Circulating CfDNA might be a biomarker for the early diagnosis of ovarian cancer.


Assessment of cell-free DNA for early detection of ovarian cancer in women. (1)


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Vol.5 No.1 – 10 : Clonal Diversity and High Prevalence of Oxa-23 among Carbapenem Resistant Acinetobacter baumannii Isolates in Egypt

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By : Nevine Fam1, Doaa Gamal1, *Dalia Salem1, Heba Dahroug1, Reham M. Wasfy1, Myriam M. Morcos2.

1Department of Microbiology, Theodor Bilharz Research Institute (TBRI), Giza, Egypt.

2Naval Medical Research Laboratories (NAMRU-3). Cairo, Egypt.

Abstract

Background and Aim: Carbapenem-resistant Acinetobacter baumannii (CRAB) is becoming a global threat especially to hospitalized patients. We aimed to address the magnitude of CRAB causing healthcare-associated infections in patients admitted to a tertiary healthcare hospital in Egypt and to study their genetic and epidemiologic diversity. Materials and Methods: Twenty-six CRAB isolates representing 48% of all Acinetobacter baumannii (A. baumannii) isolated in the study period were identified by microbiological culture methods and verified by presence of bla-oxa51. Antimicrobial susceptibility was tested by disc diffusion and MIC was determined by VITEK2 compact system. Phenotypic expression of metallo-β-lactamases (MBLs) was determined by MBL IP/IPI E-test. Carbapenemase encoding genes were identified by PCR and clonal relatedness was studied by pulsed field gel electrophoresis (PFGE) using Apa1 and PulseNet protocol. Results: All A. baumannii isolates were multi-drug resistant (MDR). Colistin and minocycline showed the highest sensitivities of 100% and 61.1%respectively. MBLs were phenotypically detected in 20/26 (76.9%) of the isolates while blaOXA-23-like was the main carbapenem resistance gene recorded in 61.5% followed by blaNDM-1-like (26.9%) and blaGES-like (7.7%).  PFGE typing showed high diversity as most of the isolates were < 80% similar. Conclusion: Carbapenem resistance among A. baumannii isolates is increasing dramatically in our geographic region. BlaOXA-23-likeis the most common gene in CRAB isolates in our hospital setting. In addition, blaNDM-1-like and blaGES-like harboring A. baumannii isolates are exhibiting a considerable spread in hospital environment in Egypt. The clonal diversity of our CRAB isolates suggests that it could be due to horizontal dissemination of mobile genetic elements rather than propagation of a certain clone.

Clonal-Diversity-and-High-Prevalence-of-Oxa-23-among-Carbapenem-Resistant-Acinetobacter-baumannii-Isolates-in-Egypt

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Vol.5 No.1 – 9 : Oxidative stress and DNA damage in Nile Tilapia (Oreochromis niloticus) as biomarkers of aquatic pollution

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By: EL-Hassan Mokhamer, Eman H. Radwan and Moataz Elsaka

Zoologlogy Department, Faculty of science, Damanhour university,Egypt

 (Email: elhassan.gaber@sci.dmu.edu.eg)

Abstract

The main purpose of this study is to evaluate the impact of heavy metals (Pb, Zn, Cu, Fe, and Cd) aquatic pollution of EL-Mahmoudeyia canal on the antioxidant enzymatic activities, GSH content and lipid peroxidation levels (MDA) in Oreochromis. niloticus muscles tissues collected from two areas EL-Mahmoudeyia canal as Polluted area and  Rosetta branch of river Nile as reference area in summer 2018 and winter 2019 as well as DNA damage was assessed in fish gills(erythrocytes) samples by applying comet assay. EL-Mahmoudeyia canal exposed to excessive of industrial effluents which impact the living organisms especially fish. The herein results showed that higher concentrations of heavy metals (Pb, Zn, Cu, Fe, and Cd) were detected in water and fish samples collected from the polluted area in comparison with the reference area, especially in winter.  The accumulation patterns of heavy metals in muscles of O. niloticus, were in the following order: Fe > Zn >Pb> Cu and Cd.The antioxidant enzymatic activities of (SOD, CAT, GPx and GST) and the lipid peroxidation biomarker MDA levels in muscles of O. niloticus collected from the polluted area were found to be significantly increased compared to that of the reference area.Meanwhile, there was a significant decrease in the GSH content levelin muscles of O. niloticus collected from the polluted area compared to that of the reference area. A significant elevation in DNA damage frequencies was observed in fish collected from the polluted areas compared with those from the reference area.These noticeable alterations in the selected antioxidant enzymatic activities in muscles of the O. niloticus go in parallel with the remarkable elevation in the levels of the detected heavy metals in water from EL Mahmoudeyia canal, as a result of pollution in these areas. This study explored the utility of the DNA damage, the altered antioxidant enzymatic activities, GSH content and MDA level as biomarkers of aquatic pollution.

 

Oxidative-stress-and-DNA-damage-in-Nile-Tilapia-Oreochromis-niloticus-as-biomark

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Vol.5 No.1 – 8 : Optimization of Microwave technique conditions for Shrimp chitin deacetylation by response surface methodology

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By : Samar Mohamed Mahdy

Department of Food Science, Faculty of Agriculture, Ain Shams University, Cairo, Egypt

E-mail: samar_mahdy@yahoo.com

Abstract

The objective of this study was to optimize chitosan production conditions from shrimp waste using the response surface methodology corresponding to the degree of deacetylation. The influence of alkaline concentration, microwave power, time and chitin concentration on the N-deacetylation of shrimp wastes chitin by microwave irradiation was investigated. Experimental conditions varied from 30 to 50% NaOH, 1125 to 2450 MHz, 6 to 18 min and 4.76 to 8.30% chitin concentration. The degree of deacetylation increased significantly with increasing time, NaOH conc. , microwave power and decreasing chitin conc. response surface analysis indicated the maximal degree of deacetylation to occur at 50% NaOH, 2400 MHz, 4.76% chitin conc. and 13 min to obtain chitosan with DDA 90.2%. 

The utility of biomarkers in the discrimination of acute coronary syndrome patients

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Vol.5 No.1 – 7 : The utility of biomarkers in the discrimination of acute coronary syndrome patients

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By : Mohamed M. Omrana*, Faten M. Zahranb, Mohamed Kadryc,

Arafa A. M. Belal c, Hesham S. Abd Alsamied 

a Chemistry Department, Faculty of Science,  Helwan University, Cairo, Egypt;

b Chemistry Department, Faculty of Science, Zagazig University, Zagazig, Egypt;

c Chemistry Department, Faculty of Science, Port Said University, Port Said, Egypt;

d Clinical Pathology Department, Faculty of Medicine, New Damietta, Al-Azhar University, Egypt;

Abstract

Background: The assessment of acute coronary syndrome (ACS) provides useful data not only for identification but also for therapeutic decisions. Objective: To develop a score based on cardiac biomarker for the early diagnosis of ACS patients. Methods: Serum concentrations of myeloperoxidase (MPO) and monocyte chemoattractant protein-1 (MCP-1) beside cardiac troponin I (cTnI) and creatine kinase MB (CK-MB) were measured in a total 200 chest pain patients represented in two groups (estimation and validation). The diagnostics value of biomarkers for discriminating ACS patients was evaluated by area under receiver-operating characteristic curve (AUC). Multivariate discriminant analysis (MDA) was used to create a predictive score. Results: A novel score depends on combination of cTnI, CK-MB, MPO and MCP-1 produces AUCs of 0.93 for discriminating ACS patients and 0.90 for acute myocardial infarction (AMI). This score correctly classified 95% of ACS patients and 93% of AMI patients with good efficiencies were 88.3% and 81%; respectively. This score had similar results in the validation study. Conclusion: Four biomarkers in combination yield a novel score help in the early and safe prediction of ACS and AMI.

Optimization of Microwave technique conditions for shrimp chitin deacetylation by response surface methodology

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Vol.5 No.1 – 6 : A Correlational Approach on the Link between Emotional Intelligence and Stress Level of Nursing Students

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By : Jordan H. Llego, RN, Ph.D., FRIHC, FRIN, FRIRes, Jennie L. Corpus, RN, FRIN

College of Nursing and Midwifery, University of Luzon 1

Perez, Boulevard, Dagupan City 2428 Philippines

(Email: llegojordan@gmail.com)

Abstract

The aim of this research study is to assess the level of emotional intelligence, the stress level and the link between the two variables. This study will use descriptive-correlational design through the use of a questionnaire; total enumeration was employed with 32 respondents. The data collected were analyzed using SPSS version 22, and mean, and Pearson-r was utilized. The mean score reveals a score of 112.29, which means that the respondents have average emotional intelligence. The mean reveals a score of 20.59, which means that the respondents are experiencing a moderate level of stress. The Pearson-r yielded a score of (r= -0.32, n=34, p=0.70) which means that there is no significant relationship between the two variables. Conclusions include: the respondents can assess others’ emotion, moreover their emotion with acceptable level of emotional control, and thus they can interact satisfactorily with other people, and positively use their emotion with a positive outlook in life; the respondents are adapting to the stress they are experiencing; and the emotional intelligence of the respondents does not have a link with their stress level.

A Correlational Approach on the Link between Emotional Intelligence and Stress Level of Nursing Students jbaar.org

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Vol.5 No.1 – 5 : Neutrophil gelatinase-associated lipocalin and oxidative stress markers based -scores to improve the diagnostic accuracy of chronic kidney diseases

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By : Toson el-SA1 , Waly S1, Omran MM2,*

1Chemistry Department, Faculty of Science, Damietta University, Damietta, Egypt
2Chemistry Department, Faculty of Science, Helwan University, Cairo, Egypt

Abstract

Background: Biomarkers can detect chronic kidney disease (CKD) in an early stage. Patients and methods: 75 individuals suffering from chronic kidney disorders were included in this study. Serum Neutrophil gelatinase-associated lipocalin (NGAL) was assayed using ELISA. Nitric oxide (NO) in addition to total antioxidant capacity (TAC) was colormetrically measured. Results: NGAL was the most efficient marker among others. Its area under receiver operating characteristic (ROC) were 0.91 and 0.80 for differentiating patients with CKD from control individuals and discriminating patients in late stages from those in early stages, respectively. The stepwise multivariate discriminant analysis (MDA) selected two novel indexes. The first is to differentiate patients in late stages from those in early stages of CKD; namely NGAL- Total antioxidant capacity (NT) score. The second is to differentiate all of these patients from those of the control subjects; namely NGAL- nitric oxide (NN) score. The AUCs of NT and NN score were 0. 84 and 0.93; respectively. Conclusion: NT and NN scores may add more in evaluation and prediction of the progression of CKD.

Neutrophil gelatinase-associated lipocalin and oxidative stress markers based -scores to improve the diagnostic accuracy of chronic kidney diseases-converted

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Vol.5 No.1 – 4 : Evaluation of the in vitro antiplasmodial activity of Millettia zechiana and its action on the evolution of anemia in albino rats.

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By : Karamoko Chérif Moustapha1,2*, Touré André Offianan2, Bla Kouakou Brice1, Tuo Karim2, N’guessan Yao Honoré1, Bidie Alain dit Philippe1

1University of Félix Houphouët Boigny, 22 PO Box 582 Abidjan 22, Côte d’Ivoire

2 Institut Pasteur of Côte d’Ivoire, 01 PO Box 490 Abidjan 01

Abstract

Background: Malaria is a parasitic infection that leads to anemia and death. Unfortunately, the upsurge of chemo-resistance prompted researchers to focus on new antimalarial drugs. Objectives: This work aimed to evaluate the antiplasmodial and antianemic activity of Millettia zechiana. Methods: the In vitro activity was assessed on clinical isolates and the standard strain of Plasmodium falciparum K1, using the SYBR green I test. Moreover, the antianemic activity was evaluated in phenylhydrazine-induced anemic albino rats. Results/discussion: The ethyl acetate and hydroethanolic extract exhibited an antiplasmodial activity with IC50s of 6.14 and 12.14 μg/mL respectively on the Plasmodium falciparum K1 strain. As for the in vivo antianemic activity, the ethyl acetate extract was the most active with better hematological reconstitution percentages. The presence of chemical compounds such as alkaloids, terpenoids, and quinonic substances in both extracts, could be responsible for their activities. Conclusion: Millettia zechiana could be a potential source for novel antimalarial drugs and might be used as an improved traditional medicine on account of its availability.

Evaluation of the in vitro antiplasmodial activity of Millettia zechiana and its action on the evolution of anemia in albino rats.-converted

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Vol.5 No.1 – 3 : Biochemical and pathological evaluation of the effectiveness of nano-targeted sono-photodynamic therapy in breast cancer

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By: Ahmed A Jasim1,2, Jubran M. Abdulrahman1,3 ,  Samir A. Abd El-kaream4,

Gihan Hosny1*

1Institute of Graduate Studies & Research, Alexandria University, Egypt.

2Al-NahrainUniversity, Biotechnology Research center, Baghdad, Iraq.

3Center of Research and Educational Studies, Ministry of Education, Baghdad, Iraq.

4Medical Research Institute, Alexandria University, Egypt.

Abstract

Background: Sono-photodynamic therapy (SPDT) is a novel cancer treatment therapy that uses the light of a particular wavelength and sound of a particular frequency to activate a light- and sonosensitive material which attaches selectively tumor cells, causing their breakdown. The present work aims to investigate biochemically and pathologically the effectiveness of using SPDT in combination with either nano-5-aminolevulinic acid (nano-ALA) or nano-Chlorophyll (nano-Chl), as sono-photosensitizers, in curing Ehrlich ascites carcinoma tumor implanted to group of mice. Methods: A total of 130 male Swiss albino mice with age 60–65 day, weighing 20 ± 2.0 g, were used in this experiment. Two sources of energy were used; namely infrared laser (IR) and Ultrasound for 3 min. When the tumor had grown to about 10 mm in diameter at day 10 after inoculation, the treatment strategy was started. Six experimental groups were investigated. Biochemical analyses were carried out to detect the serum levels for activities of oxidative stress enzymes, liver and kidney functions. Pathological analyses were carried out to evaluate the effect of different treatment modalities on tumor treatment.  Results: It was observed that SPDT utilizing nano-ALA or nano-Chl ameliorated the activity levels of the studied enzymes in mice bearing tumor lesions, indicating the safe and non-toxic treatment modality. Pathological evaluation showed that targeted SPDT caused almost complete destruction of cancer cells. Conclusion: The results suggest that nano-ALA or nano-Chl could be used as novel and safe nano-materials with great potential as effective drug delivery system in cancer-targeted SPDT.

Biochemical and pathological evaluation of the effectiveness of nano-converted

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