ORIGINAL_ARTICLE
MONOCYTE-DERIVED DENDRITIC CELLS ISOLATION AND PHENOTYPIC CHARACTERIZATION
Dendritic cell (DC) is the most powerful inducers and regulators of immune responses, responsible for interaction within the immune system. The ability of DC to induce or regulate/suppress immune responses led to attention in their immunotherapeutic use in various disease types. The aim of the presented study was to generate in vitro CD14+ enriched peripheral blood monocytes (PBMCs) following the culture with human monocyte-derived dendritic cell serum-free differentiation media over ten days with phenotypical analysis and morphological identification for functional studies. In vitro, peripheral blood of human donors samples were collected for the purification of blood CD14+ monocytes that represent the most common origin for DC precursors based on ficoll density gradient separation and human monocyte-derived dendritic cell enriched differentiation medium. After PBMCs isolation, the cell viability, cell yield were determined, the monocyte-derived dendritic cell surface marker expression was detected by flow cytometric analysis after staining with specific fluorescence-conjugated monoclonal antibodies. in vitro culture and differentiation of human monocytes into DCs and their subsequent maturation into mature DC constitute a critical first step for different downstream analysis ranging from an understanding of both human DC biology and function to their use in clinical diagnostics and therapeutic design for different disease.
https://ajps.journals.ekb.eg/article_118369_f3bc381b4480a26041274e13ac55b2be.pdf
2020-09-01
1
11
10.21608/ajps.2020.118369
Monocyte
Dendritic cell
APCs, antigen-presenting cells
Ficoll density gradient separation
mohamed
farag
mohamed.farag@azhar.edu.eg
1
Department of Botany and Microbiology, Faculty of Science, Al-Azhar University, ‎Cairo, Egypt
AUTHOR
ORIGINAL_ARTICLE
EVALUATION OF NATURAL KILLER CELLS USING DIFFERENT TECHNIQUES
Natural killer (NK) cells mediate spontaneous cytotoxicity toward abnormal and tumor cells and rapidly secrete numerous cytokines and chemokine`s which promote subsequent adaptive immune responses. The presented study aimed to compare between different methods for NK evaluation and determination. Ten samples of healthy individuals were collected from Al-Zahraa University hospital and evaluated for phenotypic identification for NK cells. Isolation procedure of NK was performed in Clinical Pathology Department, Hematology Unit, Flow-cytometry lab, Al-Azhar University. Protocols started with isolation of peripheral blood mononuclear cells (PBMCs) from fresh human whole blood depending on Ficoll-Paque density gradient centrifugation. Other protocol depended on flow cytometry sorting technique. All experiments were carried out in accordance with the approved guidelines. Sorting procedure using FACS Calibur machine yielded 64.97 % purity before cells were cultured, while mixed Ficoll separation of whole blood showed purity 22.74%. Through our study, it is clear to us that the percentage of purity of NK cells using the flow-cytometry sorting technique is much higher than the percentage of purity of NK cells using the separation method dependent on Ficoll-Paque density technique, Therefore, through our study, we see that it is better to rely on the flow-cytometry sorting technique using the flow cytometry device and CD56 +CD16 and CD3 antibodies in order to obtain the best purity of the NK cells (CD56+ CD16 positive /CD3 negative population) for use in medical purposes and laboratory research.
https://ajps.journals.ekb.eg/article_118370_fbfea72af255ea1c30d9847d357fdc48.pdf
2020-09-01
12
22
10.21608/ajps.2020.118370
FACS Calibur
Natural Killers
PBMCs
Ficoll
Mohamed
Selim
mohamedselim@azhar.edu.eg
1
AUTHOR
ORIGINAL_ARTICLE
BIODETERIORATION OF CYPERMETHRIN BY LOCAL ISOLATE ISOLATED FROM EGYPTIAN PETROLEUM CONTAMINATED SOIL
Cypermethrin is a widespread pyrethroid pesticide and is an environmental pollutant because of its toxicity and persistence. Biodegradation of such chemicals by microorganisms may provide a cost-effective method for their detoxification. Sixty-two cypermethrin degrading bacteria were isolated from Egyptian contaminated soil and water samples. The most potent bacterial strain was identified as Ochrobactrum intermedium SP9 based on 16S rRNA gene sequence analysis, as well as several morphological, physiological and biochemical characteristics examinations on classical level. The maximum biodegradation of cypermethrin by bacterial strain SP9 was achieved at 35°C and pH 7.0, and the degradation rate reached 69.1% within 8 days under the optimal conditions. Hence, the Ochrobactrum intermedium SP9could potentially be used in the future for bioremediation of cypermethrin contaminated soil.
https://ajps.journals.ekb.eg/article_118372_2316005a7e5430c02afd0d676f7e79e8.pdf
2020-09-01
23
38
10.21608/ajps.2020.118372
cypermethrin
Ochrobactrum intermedium
biodegradation
pesticide
contaminated soil
hazem
Attya
saad_hassan@azhar.edu.eg
1
Department of Botany and Microbiology, Faculty of Science, AL-Azhar University, Cairo, Egypt
AUTHOR
ORIGINAL_ARTICLE
REVIEW ARTICLE; ANTICANCER ACTIVITIES OF SOME FUSED HETEROCYCLIC MOIETIES CONTAINING NITROGEN AND/OR SULFUR HETEROATOMS
Cancer cells often have a high demand for antiapoptotic proteins in order to resist programmed cell death. Heterocyclic compounds present themselves as a fundamental division of organic chemistry. Most clinically effective antitumor agents are inhibitors of DNA, RNA or protein synthesis, enzyme inhibition and/or interfere with the metabolism of other cell components, frequently lacking or displaying poor selective antitumor activity. The majority of heterocycle compounds and typically common heterocycle fragments present in most pharmaceuticals currently marketed, alongside with their intrinsic versatility and unique physicochemical properties have poised them as true cornerstones of medicinal chemistry. The S-heterocyclic core (Thiophene) has been reported to possess significant importance in various fields from medicinal chemistry. Many diverse biologically active products were prepared, several of which exhibited antimicrobial, analgesic, anti-inflammatory, antioxidant, antitumor, local anesthetic, anticoagulant and antithrombotic activities. Many N-heterocyclic compounds that are broadly distributed in Nature, possess physiological and pharmacological properties and are constituents of many biologically important molecules, including many vitamins, nucleic acids, pharmaceuticals, antibiotics, dyes and agrochemicals, amongst many others. The base pairs of DNA and RNA (Guanine, cytosine, adenine, and thymine) are also made up of N-heterocyclic compounds, namely purines, pyrimidines, etc. These nitrogen-containing heterocyclic molecules with distinct characteristics and applications have gained prominence in the rapidly expanding fields of organic and medicinal chemistry and the pharmaceutical industry. Finally, many of these compounds have higher activity against cancer cells using the standard MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)assay.
https://ajps.journals.ekb.eg/article_118375_61d0fdf9ac43ad7bbcd14d9a1e5ac335.pdf
2020-09-01
39
54
10.21608/ajps.2020.118375
Heterocyclic compounds
breast cancer
Hepatocellular carcinoma
Nitrogen and/or Sulfur-based heterocycles
Asmaa
Omar
eladlkhaled74@azhar.edu.eg
1
Department of Chemistry, Faculty of Science, Menoufia University, Shebin El-Koam, Egypt
AUTHOR
ORIGINAL_ARTICLE
THE ROLE OF SERUM OSTEOPONTIN LEVELS IN EGYPTIAN WOMEN WITH POSTMENOPAUSAL OSTEOPOROSIS
Background: Postmenopausal osteoporosis (PMO) is a debilitating disease induced by estrogen deficiency characterized by bone loss and bone micro-architecture degradation contributing to fragility fractures. Osteopontin (OPN) has been found to be involved in bone turnover by activating the resorption process. Objectives: This study aim to evaluate the relationship between serum OPN levels and bone mineral density (BMD) for identification of female individuals at high risk of osteoporotic fractures. Methods: A total of eighty eight postmenopausal women (PMW) were recruited in this study. Densitometry results addressed cases of the study into 3 groups: thirty with osteoporosis, thirty with osteopenia and twenty eight PMW without complications as a control group. Serum OPN was measured by using Enzyme-Linked immunosorbent assay (ELISA) kits and BMD was obtained at lumbar spine and femoral neck by dual-energy X-ray absorptiometry (DEXA). Results: Osteopontin levels were significantly higher in osteoporotic group compared to osteopenic and control groups (P ˂ 0.001). Negative correlation was found between BMD and OPN in osteoporotic group. By using the Receiver Operating Characteristic (ROC) curve, the cutoff level of ≥ 10.294 ng/mL for OPN was chosen for suggesting osteoporosis which yield a sensitivity of 66.67 % and specificity of 96.43 %. Also, there were a significant increase in serum ALP levels in osteoporotic and osteopenic groups compared to control group while serum Ca failed to show a significant difference between the studied groups. In addition, serum TAC showed a significant decrease in osteoporotic group compared to control group. Conclusion: These findings suggest that the circulating OPN might play role in the pathophysiology of PMO. Its blood concentration can be used as a sensitive monitoring indicator for the early detection of osteoporosis independently of BMD.
https://ajps.journals.ekb.eg/article_118376_9d06e0af85ab3d9948dffec37c04c8b7.pdf
2020-09-01
55
71
10.21608/ajps.2020.118376
Bone turnover
Estrogen
Osteopontin
Osteoporosis
Amira
Atta
amira-ahmed@eru.edu.eg
1
Department of Biochemistry, Faculty of Pharmacy, Egyptian Russian University
AUTHOR
ORIGINAL_ARTICLE
A NEW TRIAL FOR BIOFORMATION OF ANTIMICROBIAL AGENT CONTROLLING MULTI DRUG RESISTANT MICROORGANISM
In the present investigation, a trial was done to find a new antimicrobial agent producing microbe from soil microbiota of local habitats to control the problem of multiple drug resistance. The term of Antimicrobial resistance (AMR) is used to describe microorganisms that can resist the effects of drugs and chemicals designed to kill them. Seventy six actinomycetes isolates were isolated from fifteen soil samples different localities in Egypt were primary screening for antimicrobial activity by agar plug diffusion method against test microorganisms. Sixteen isolates were selected for secondary screening in small scale submerged fermentation system and assayed against pathogenic tested microorganisms using agar well diffusion method. Among of these isolates tested, the isolate (S1SHA1) showed the highest antimicrobial activity against pathogenic test organisms. This isolate was identified as Streptomyces griseoplanus by morphological, physiological, biochemical characters and 16s rRNA gene sequence. Physical and nutritional factors affecting activity of antimicrobial agent were studied. The results showed that, optimum activity of antimicrobial agent achieved with pH 7, incubation temperature 28 C, for 7 days, at 150 rpm agitation, carbon and nitrogen source starch 1.5% and potassium nitrate 0.4%, as well as phosphorus 2 g/l and NaCl at concentration of 1 %. Antimicrobial agent from batch culture was subjected to extraction and purification processes using ethyle acetate and preparative TLC, respectively. Determination of minimum inhibitory concentrations (MIC) and Mode of action of antimicrobial agent produced by S. griseoplanus (S1SHA1) on the test microbial strains using Transmission Electron Microscopy (TEM). Cytotoxic studies showed that no cytotoxic effects were observed for the compound when tested even at high concentrations
https://ajps.journals.ekb.eg/article_118377_917e9bece8568dac11ba59fcd3ee087e.pdf
2020-09-01
72
95
10.21608/ajps.2020.118377
Pathogenic microorganism
antimicrobial agent
multidrug resistant
Cytotoxicity
Mahmoud
Elaasser
mmelaaser@azhar.edu.eg
1
Assistant professor of Microbiology - The Regional Center for Mycology and biotechnology, Al-Azhar University, Cairo
AUTHOR
ORIGINAL_ARTICLE
PRELIMINARY CHARACTERIZATION AND IDENTIFICATION OF GRAM POSITIVE HEMOLYSIS BACTERIA
One of the most important pathogens that threaten human health all over the world is Staphylococcus spp. characterization and identification of such pathogen considers a useful tool to control of some serious problems resulting by these bacteria. Therefore one hundred and twenty samples including blood, urine, abscess, semen, pus, sputum, ears, vaginal and spit swabs were collected from patients of Tanta University Hospital and outpatient clinics. A total of 126 Gram positive bacterial isolates were obtained from these clinical specimens. Of these isolates 107 bacterial strains were identified as Staphylococcus spp and 15 strains identified as Streptococcus spp in addition to 4 strains were identified as Enterococcus spp, preliminary identification was conducted on the basis of colony characteristics such as Gram staining, pigment production, hemolysis, catalase activity, coagulase test and fermentation of manitol sugar. Out of these strains, 30.95% have the potency to make alpha hemolysis while 30.95% have the ability to make beta hemolysis and 38.09% posse the capacity to make gamma hemolysis on blood agar medium. Beta hemolysis Staphylococcus spp were selected for study of some virulence factors on basis of coagulase production in which 17.24% of Staphylococcus spp were coagulase positive and 82.76% were coagulase negative Staphylococcus spp. Studying the susceptibility pattern of these strains to some commercial antibiotics was carried out. Further future studies are recommended to investigate the effect of some natural compounds on gene regulation that responsible for hemolysis process.
https://ajps.journals.ekb.eg/article_118378_e541964c113e5b4c015c882cbd1021e7.pdf
2020-09-01
96
109
10.21608/ajps.2020.118378
Hemolysis
Gram positive bacteria
Staphylococcus
Coagulase production
Ahmed
Almwafy
ahmed.tarek.201@azhar.edu.eg
1
Department of Botany and Microbiology, Faculty of Science (Boys), Al-azhar University, Cairo, Egypt.
AUTHOR
ORIGINAL_ARTICLE
PHYTOCHEMICAL AND BIOLOGICAL STUDIES ON SOME MEDICINAL PLANTS
Biological studies on thirteen ethanolic plant extracts (Rosmarinus officinalis ,Ocimum basilicum , Moringa oleifera, Zingiber officinale, Curcuma longa, Nigella sativa ,Cinnamomum verum ,Salvia officinalis, Lepidium sativum, Foeniculum vulgare, Anethum graveolens, Ficus benghalensis and Cinnamomum camphora )revealed that four of them (Rosmarinus officinalis , Zingiber officinale, Cinnamomum verum and Cinnamomum camphora) were the best active against two bacterial species ; E. coli , S .aureus and one fungus species C. albicans. Also, their synergistic effects against E. coli, S. aureus and C. albicans were studied .So, the phytochemical studies were completed on these four plants . This study aimed to evaluate the chemical composition of the best active plant extracts. The chemical major content of Rosmarinus officinalis was eucalyptol (7.48%) , Zingiber officinale was gingerol (12.73%) , Cinnamomum verum was (E)- cinnamaldehyde (25.55 % ) and Cinnamomum camphora was eugenol ( 27 .35% ) . The minimum inhibitory concentration (MIC) values varied from 0.625 to 2.5 mg/ml, for the S. aureus (gram positive bacteria) affected by Rosmarinus officinali, Zingiber officinale, Cinnamomum verum and Cinnamomum camphora. Respectively. C. albicans was the most effective microorganism by Cinnamomum verum and the least effective microorganism by Rosmarinus officinalis and Cinnamomum camphora. ethanolic extracts, as MIC ranged from 0.15 to 1.25 mg/ml.
https://ajps.journals.ekb.eg/article_118379_5217b87f66819710cff47f3b6e2773d7.pdf
2020-09-01
110
134
10.21608/ajps.2020.118379
Escherichia coli
Staphylococcus aureus
Candida albicans
Rosmarinus officinalis
Zingiber officinale
Cinnamomum verum
Cinnamomum camphora
antimicrobial activity
Minimum inhibitory concentration (MIC)
Amina
El-Kelany
aminamohammed.2059@azhar.edu.eg
1
Department of Botany and Microbiology, Faculty of Science, Al-Azhar, University, Girl Branch, Cairo, Egypt.
AUTHOR
ORIGINAL_ARTICLE
PROTECTIVE EFFECT OF QUERCETIN AGAINST STATINS INDUCED-HEPATOTOXICITY IN CELL LINE
Objective: Statins are the most prescribed lipid lowering agents and consequently they prevent obstructive cardiovascular events in the world. Severe adverse effects of statins, involving myopathy and hepatotoxicity, sometimes limit their usage as lipid lowering agents. We now investigate the toxicity of statins and prove the protective effect of quercetin against statins toxicity in cell line. Methods: Human hepatocellular carcinoma cells HepG2 were used in this study were cultured at 37℃ in 5% CO2, in Roswell Park Memorial Institute medium (RPMI 1640) and were supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 mg/mL streptomycin. HepG2 cells were cultured with some of statins as Atorvastatin, Simvastatin and Rosuvastatin with 10µM concentration with and without pretreatment with 10, 20, 40µM quercetin 4 hours before the addition of statins, then cell line was incubated for 24 and 48 hours. Toxicity of statins was determined by cell viability assay (MTT assay), ALT&AST level assay, lipid peroxidation assay by Malondialdehyde (MDA) and Immunofluorescence staining assay using DAPI. Results: ALT&AST levels were significantly increased after the addition of statins in HepG2 cells when compared with the control group (DMSO 0.1%), and ALT&AST levels were significantly decreased with pretreatment of the cells with quercetin 4 hours before the addition of statins in a dose-dependent manner when compared with statin group. Cell viability percentage (MTT concentration) was decreased significantly after the addition of statins to HepG2 cells when compared with the control group, and MTT concentration was significantly increased with pretreatment of the cells with quercetin 4 hours before the addition of statins in a dose-dependent manner when compared with statin group. MDA concentrations increased significantly after the addition of statins to HepG2 cells when compared with the control group, and decreased when the cells were pretreated with quercetin 4 hours before the addition of statins in a dose-dependent manner when compared with statin group. Statins cause cellular oxidative damage by liberating reactive oxygen species, and the cellular damage was prevented when the cells pretreated with quercetin 4 hours before addition of statins. Conclusions: We found that, quercetin shall protect HepG2 cells from statins-induced hepatotoxicity with 10, 20, 40µM concentrations without significant difference between 20 and 40µM concentrations, and may be developed as a therapeutic agent for possible statins toxicity.
https://ajps.journals.ekb.eg/article_118380_04257a326386411ba140fec4962c7449.pdf
2020-09-01
135
151
10.21608/ajps.2020.118380
statins
Drug induced liver injury (DILI)
Quercetin
MDA
MTT
Immunofluorescence assay
DAPI staining
Mohamad
Kamal
mohamadkamal.202@azhar.edu.eg
1
Biochemistry, Faculty of Pharmacy(Boys), Alazhar
AUTHOR
ORIGINAL_ARTICLE
ISOLATION AND CHARACTERIZATION OF CELLULOSE NANO FIBER PRODUCING BACTERIAL STRAIN FROM FERMENTED FRUITS
A bacterial cellulose (BC) producing strain isolated from fermented fruit. Twenty BC producing bacteria were isolated from each the isolation sources (fermented fruits). The most potent strain was identified to be Komagataeibacter xylinus SB3.1 based on several morphological characteristics, biochemical tests and 16srRNA. The Komagataeibacter xylinus SB3.1. was produce BC within pH 4–9 and exhibit maximum BC production (2.4 g/L) at pH 6 in under static conditions for 7 days. The structure of BC produced from the tested strains was assayed by scanning electron microscope it was revealed the diameter of thin ribbons ranged from 34.34 nm to 39.16 nm and exhibits higher porosity (81.5%).In comparison with the specimen from model BC producer, Gluconacetobacter xylinus 10245. Based on these analyses, the isolated Komagataeibacter xylinus SB3.1 can efficiently produce BC, which can be applied for industrial manufacturing with potential features.
https://ajps.journals.ekb.eg/article_118381_378b122a551d35d9f63ef38076a9ee38.pdf
2020-09-01
152
163
10.21608/ajps.2020.118381
bacterial cellulose
Komagataeibacter xylinus , Nanocellulose , Acetobacter and fermented fruits
Ahmed
Alemam
ahmed.alemam@azhar.edu.eg
1
Department of Botany and microbiology, Faculty of science, Al-Azhar University, Cairo, Egypt
AUTHOR
ORIGINAL_ARTICLE
BIOSYNTHESIS, CHARACTERIZATION AND ANTIMICROBIAL ACTIVITY OF IRON OXIDE NANOPARTICLES SYNTHESIZED BY FUNGI
In the present study, extracellular synthesis of iron oxide nanoparticles (FeNPs) was achieved byAspergillus flavus isolate D05.The biosynthesized iron oxide nanoparticles have been widely favored because of biodegradablity, low toxicity and highly reactive surfaces. IONPs were synthesized through the reduction of aqueous Fe3+ ions. The obtained iron oxide nanoparticles were characterized by UV-vis spectroscopy, Fourier transforms infrared spectroscopy(FTIR), and Transmission electron microscope (TEM). TEM image of iron nanoparticles synthesized byAspergillus flavus sp. showed 28-33 nm sized particles.
https://ajps.journals.ekb.eg/article_118382_f1d9dd6e4dda37ad55941fffffc3e585.pdf
2020-09-01
164
179
10.21608/ajps.2020.118382
Nanoparticles
Green synthesis
Aspergillus flavus
iron oxide NPs
antimicrobial activity
Nagwa
Sidkey
nagwasidkey@azhar.edu.eg
1
Department of Botany and Microbiology, Faculty of Science, Al-Azhar University, Nasr City, Cairo, Egypt
AUTHOR