Biotechnology Research Center, Al-Nahrain University
Molecular Immunology and Animal Cell Culture
Suhair Dakhil Neamah, Hayder Z. Ali, and Mohammad M.F. Al-Halbosiy University of Baghdad College of Science
Visceral leishmaniasis (VL), the second-most-serious parasitic illness after malaria, is currently endemic in more than 88 countries. Need for new anti-leishmanial compounds is currently being taken into consideration by researchers due to resistance and lack of effective vaccinations. This research was conducted to find out more about the effect of artemisinin (ART). ART was examined in vitro promastigotes stages and ex vivo amastigotes stages of the Iraqi strain of Leishmania donovani in U937 cell line after 24, 48 and 27 hours using MTT assay. In addition, the level of macrophage nitric oxide (NO) was measured using Griess assay in U937 cell line. The results of promastigotes viability percentage were less than 50 %, where the IC50 was 43.8, 37.9 and 36.6 µM respectively. Similar cytotoxic effect of ART was observed against amastigote forms, where the cell viability was 36.11, 40.2 and 37.4 µM respectively, after the three times of follow up.. Furthermore, by using Griess assay the level of nitrite in infected macrophages was measured. The results after 48hr were 6.58 in high concentration and 1.22 in lower concentration. However, considering the absence of a significant difference, the observed concentrations are still higher than the control when compared to Griess assay.. These findings demonstrated that ART has a cytotoxic effect on both forms of L. donovani. It also increased iNOS production in infected macrophages which can define artemisinin therapeutic efficacy against L. donovani infection.
Mustafa Attiyah Hadid, Mohammad M. F. Al-Halbosiy, and Abdulwahid B. Al-Shaibani Korean Society for Microbiology and Biotechnology
Hiba A.S. Alhadidi, Safaa A. S. Al-Qaysi, and Mohammad M. F. Al-Halbosiy Microbiology and Biotechnology Letters Korean Society for Microbiology and Biotechnology
Raghda Makia, Khulood Al-Sammarrae, Mohammad Al-Halbosiy, Mohammed Al-Mashhadani, , , , and CMV Verlag
Background Normally happening substances like flavonoids are regarded as active candidates for the treatment and prevention of cancer The purpose of this study was to see how Iraqi E. arvense total flavonoid affected cell lines biologically and human lung fibroblast normal cell line (WISH). Methods Plant powder was extracted by reflex apparatus, then thin-layer chromatography (TLC) was used to determine total flavonoids. Cytotoxicity assay (MTT) was used to determine the cytotoxic activity of the prepared plant against human breast cancer (MCF-7), cells human cervix cancer (HELA), human colon cancer (Caco-2) and human lung fibroblast normal cell line (WISH). Results The flavonoids Rutin, Quercetin, Kaempferol, and luteolin were detected using the Thin Layer Chromatography (TLC) technique. In contrast to the negative control, the extract inhibited cell growth to a highest of 82.158% for MCF-7 and 61.360% for Caco-2 at the concentration (100 µg/ml), and (54.880%) for Hela cell line at the concentration (100 µg/ml). In addition, the concentration (6.25 µg/ml) of total flavonoid extract produced a decrease in the growth of the normal WISH cell line to reach (1.094%). Conclusion Equisetum arvense contain high amounts of flavonoids, the qualification of some flavonoids compounds was detected using TLC. The total flavonoids showed significant cytotoxic activity against various types of cancer cell lines and normal cell line in vitro, the antitumor activity was highly efficient in a dose and cell type dependent manner.
Suhair Dakhil Neamah, H. Z. Ali and M. M. Al-halbosiy Annals of parasitology
Leishmaniosis is a parasitic infection spreads to humans by sand flies. Over 20 different species of Leishmania are responsible for the disease, which infect over 14 million people around the world. Chemotherapy is one of the most effective treatments for leishmaniosis, however it is restricted by the high cost and/or toxicity. In this study, the possible effect of artemisinin (ART) was detected on intracellular amastigotes of Iraqi strain of Leishmania donovani in ex vivo condition in U937 macrophage cell line. Gene expression of inducible nitric oxide synthase (iNOS) in U937 macrophage was investigated, before and after treatment with artemisinin. Kinetic result by real-time PCR demonstrated that the iNOS expression folding reached the maximum at concentration of 500 μM after 24 hours, at 750 μM after 48 hours and at 1000 μM after 72 hours, which was 56, 11, and 6, respectively. The copy number of iNOS gene expression was also significantly higher in treated infected U937 cells compared to both non-treated-infected cells and intact macrophages, under different concentration of ART along the three times of follow-up. Moreover, stained macrophages with fluorescent DAPI proved that the percentage of intracellular infective amastigotes was decreased to the minimum in treated U937 cells, in comparison to non-treated cells. The minimal amastigote-invasion percentage was recorded at 1000 μM, which was 26%, 27%, 21% compared to 61%, 87%, 75% in untreated cells after 24, 48, 72 hours respectively. These findings demonstrated ART positive efficacy against iNOS expression and this compound can be further studied as novel therapeutic rather than toxic available chemotherapies.
Safaa A. S. Al-Qaysi, Halah Al-Haideri, Sana M. Al-Shimmary, Jasim M. Abdulhameed, Othman I. Alajrawy, Mohammad M. Al-Halbosiy, Tarek A. A. Moussa, and Mohamed G. Farahat Journal of Microbiology and Biotechnology
Levan is an industrially important functional biopolymer with considerable applications in food and pharmaceutical fields owing to its safety and biocompatibility. Herein, levan type exopolysaccharide (EPS) produced by Pantoea agglomerans ZMR7 was purified by cold ethanol precipitation and characterized using TLC, FTIR, 1H, and 13C NMR spectroscopy. The maximum production of levan (28.4 g/L) was achieved when sucrose and ammonium chloride were used as carbon and nitrogen sources, respectively, at 35 °C and an initial pH value of 8.0. Some biomedical applications of levan like antitumor, antiparasitic, and antioxidant activities were investigated in vitro. The results revealed the ability of levan at different concentrations to decrease the viability of rhabdomyosarcoma (RD) and breast cancer (MDA) cells compared with untreated cancer cells. Besides, levan appeared to have high antiparasitic activity against the promastigote of Leishmania tropica. Furthermore, levan had strong DPPH radical scavenging (antioxidant) activity. These findings suggest that levan produced by P. agglomerans ZMR7 can serve as a natural biopolymer candidate in pharmaceutical and medical fields.
Shaymaa Rajab Baqer, Abdulkareem Alsammarraie, Mahasin Alias, Mohammad Al-Halbosiy, and Amaal Sadiq Baghdad Science Journal College of Science for Women
Titanium dioxide nanotubes were synthesized by anodizing Ti sheets in the ethylene glycol solution and were covered in Pt nanoparticles onto the surface of TiO2NTs using electrodeposition method from using five derivatives of Mannich base Pt complexes which have been used as precursor of platinum. The mean size, shape, elemental composition of the titanium dioxide nanotubes and platinum deposited on the template were evaluated by different techniques such as field emission scanning electron microscope (FE-SEM), transmission electron microscopy (TEM), X-ray diffraction pattern (XRD), and energy dispersive X-ray (EDX) technique. From all these analyses, the TiO2NTs prepared and Ptnanoparticles deposited on it were identified. The diagnoses proved that all the Pt nanoparticles have a size less than 50 nm. The MCF-7 cancer cell lines and WRL68 normal cell lines were treated with concentration 800, 400,200,100, 50, 25, 12.5µg\\ml of TiO2NTs and Pt\\TiO2NTs(1) and (2) for 48hours using MTT assay.IC50 and inhibition rate were calculated. The result shows that the Pt\\TiO2NTs have more inhibition effect on cancer cell lines than TiO2NTs array.
M. F. Al-Halbosiy, Hayder Z. Ali, G. M. Hassan and F. Ghaffarifar
Visceral leishmaniosis is one of the most fatal old-world neglected disease with estimated 90 thousand worldwide cases emerge each year. In Iraq, the cutaneous and visceral form are endemic but available chemotherapies are either toxic with diverse side effects, expensive available drugs or parasite resistant is arising. Artemisinin (ART) is a semi-synthetic compound which proved its effectiveness against protozoan parasites, such as malaria and Leishmania. In this study, the efficacy of different concentrations of pure artemisinin was screened in vitro against promastigotes and axenic amastigotes by MTT assay after 24, 48 and 27 hours follow up. In addition, the infectivity ability and number was investigated of intra-cellular Leishman bodies in treated murine peritoneal macrophages after 24 and 48 hours ART treatment. The results verified ART efficacy against the promastigotes and axenic amastigotes viability with IC50 measured after 24, 48- and 72-hours treatment. Infectivity percentage of murine macrophages and parasite burden were significantly reduced in treated cells. These findings indicate the leishmanicidal activity of ART against the Iraqi isolate of L. donovani and further in vivo study is recommended for assigning ART as a natural anti visceral leishmaniosis compound.
May Ridha Jaafar, Yaseen Ismael Mamoori, and Mohammad Mahmoud Al-Halbosiy Africa Health Research Organization
Cutaneous leishmaniasis is a tropical disease caused byLeishmania parasiteand vaccination is the best way to control it.In this study, the truncatedHaemoglobin receptor (HbR-N) gene of Leishmaniatropicawas cloned and expressed in Escherichia coli.The sequence of this gene was codon-optimized, synthetically synthesized and amplified using specific primers. pTYB21 plasmid as well as the amplified gene were digested bySapI and PstI and ligated using T4 DNA ligase. The recombinant plasmid was used to transform DH5α Escherichia coli. Transformants were confirmed using colony PCR, digestion of the plasmids using PvuII enzyme as well as DNA sequencing. The recombinant plasmid was used to retransform the T7 express E. coli and the transformants were confirmed using colony PCR. The cell carrying the plasmid was used to express HbR-N protein. The gene expression was analysed by SDS-page polyacrylamide gels. PCR has successfully amplified the HbR-N gene with a molecular size of 398 bp.Transformants carrying the pTYB21_HbR-N plasmid were obtained and designated as MYM1. DNA sequencing showed 100% similarity to HbR gene. The later plasmid was used to retransform T7 express E. coli, several transformants were obtained and they designated as MYM2. The MYM2 bacterial cell was used to express the gene. Finally, SDS-page polyacrylamide gel showed protein bands of the fused protein (intein-HbR-N) with a molecular size of about 70 kDa. In this study, the HbR-N gene was cloned and expressed in E. coli and could be used as a potential vaccine against several Leishmania spp.
Mohammad Mahmoud Farhan Al-Halbosiy, Zainab Yaseen Mohammed Hasan, Farooq Ibrahem Mohammad, and Baraa Abdulhadi Abdulhameed Iraqi Journal of Science University of Baghdad College of Science
The present study focuses on the biological strategy for treating cancer and parasitic infections, such as leishmaniasis antiparasitic activity, forthe crude alcoholic extract ofFicus leaves and their extracted total flavonoids with a comparison between their effects.The flavonoids were extracted from the leaves of the mature Ficuscaricausing the reflux extraction method. Total flavonoids were detected qualitatively by TLC techniquewhich demonstratedthat the plant was rich indifferent flavonoids, especially Rutin, Quercetin, Kaempferol, luteolin and others. Quantitatively, the plant total flavonoids content was 337.3 mg / 100 g fig leaves calculated as rutin.The biologicaleffects of the crude and purified total flavonoid on cell lines (L-20B and MCF7) and two parasites (Leishmaniatropica ,Leishmaniadonvani) were investigated. Maximum growthinhibition rates forthe total flavonoids onthe cell linesL20B andMCF7 reached 43 % at the concentration of 0.169 mg/ml and 28% at the concentration of 2.7 mg/ml, respectively, in comparison with the negative control. The ethanoliccrude extract had a low effect on L20-B cell line, while the inhibition rate forMCF-7 cell linereached 34% at a concentration of 0.084 mg/ml. For Leishmaniatropica,the total flavonoid and crude plant extractcaused maximum inhibition rates of 48% and 56%, respectively,at a concentration of 2.7 mg/ml for both. Cytotoxicity valueon Leshmaniadonovani was 20% for the crude extract at 1.35 mg/ml concentration, whereas it was11% for the total flavonoids at a concentration of 0.169 mg/ml. In conclusion, the differences in anticancer and anti-parasitic activitiesareattributed to differentcompounds present in each extract.
Ban H. Adil, Mohammad M. Farhan Al-Halbosiy, and Hamid H. Murbat AIP Publishing
This study illustrates in vitro effect of cold atmospheric plasma (CAP) on the treatment of Leishmania. In addition, the study evaluated the effect of drug treatment (pentostam) and the combination treatment of pentostan and CAP at different doses and incubation time 24 h in order to assess the most effective treatment. The duration of the cold plasma doses was 1 min, 2 min and 3 min, while the pentostam doses were 2.5 µg/ml and 5 µg/ml. The combinations therapies included combining the three cold plasma doses with the pentostam doses to test 6 combinations of treated in vitro. The maximum growth inhibition was given by combination treated 46% and 44% for donovani and tropica leishamnia respectively; these results give an indication that the cold atmospheric plasma is a promising treatment of this parasite that threats people all over the world.
Saja H. Ali, Ghassan M. Sulaiman, Mohammad M. F. Al-Halbosiy, Majid S. Jabir, and Anaheed H. Hameed Informa UK Limited
Abstract Hesperidin, as a flavonone, is recognized as promising anti-inflammatory, antioxidant, and anticancer agent. Its poor bioavailability is crucial bottleneck for therapeutic efficacy. To enhance the stability and bioactive potentials, hesperidin -PLGA-Poloxamer 407 was successfully prepared to minimize or overcome problems associated with hesperidin absorption. The characteristics of nanohesperidin were testing by in vitro dissolution study, XRD, FTIR, PSA and SEM. Antioxidant effects of nanohesperidin were studied. The structure–activity relationship analysis with antioxidant pharmacophore has been performed by using density functional theory method and quantum chemical calculations. The structural properties were investigated using Becke three-parameter hybrid exchange and the Lee–Yang–Parr correction functional methods. Nanohesperidin was found to decrease the H2O2 activity–induced DNA instability. Blood compatibility on human erythrocytes was confirmed by haemolytic and in vitro toxicity assessments. The in vitro anticancer activity of nanohesperidin towards MCF-7 cells using various parameters was carried out. The nanohesperidin was found to exert cell growth arrest, activated DNA fragmentation and induced apoptotic cell death through caspase-3 and p53-dependent pathways. These findings showed that nanohesperidin play an important role in its anticancer effects, suggesting might be used for clinical trials and can represent driving formulation for novel chemotherapeutic agents.
Mohammed A. Al-Dolaymi, Khalid F. Al-Rawi, Firas T. Al-Shamary, and Mohammad M.F. Al-Halbosiy Universitas Gadjah Mada
Bosom malignancy is the most regularly analyzed disease and the imperative reason for growth-related passing among ladies, accounting for 23% of all new tumor cases and 14% of tumor passing's. L-Ascorbic acid, commonly known as vitamin C is well-known in chemistry since long back. It has tremendous medical applications in several diseases. Therefore, in this paper five concentrations of complex cis-[Pt(Asc)(NH3)2] where Asc=L-ascorbic acid derivative on MCF-7 cell line to detect the changes in five cellular parameters (nuclear intensity, mitochondrial membrane potential, valid cell count, cytochrome C, and membrane permeability) after exposure with 24 h are investigated. The results showed that 400 μg/mL has the highest significant effect on the five parameters (nuclear intensity, mitochondrial membrane potential, valid cell count, cytochrome C, and membrane permeability) when compared with Doxorubicin 20 μM (substance used as anti-cancer) which represent the positive control. Also, the 200 μg/mL showed results close to those of the untreated cells which represent the negative control (-ve) with a very few significant differences.
Hadeel A. Nasser, May K. Ismael and Mohammed M. F. Al. Halbosiy University of Baghdad College of Science
Over the last few years the role of microorganisms in the pathogenesis of atherosclerosis has been widely discussed. Advance in basic science have established a fundamental role for inflammation immediating all stages of cardiovascular diseases. Chlamydia pneumoniae activates immune cells to produce cytokines such us TNF-α that are important contributor to atherosclerosis. All blood samples were assayed for molecular detection of Chlamydia pneumonia e by using conventional polymerase chain reaction (PCR) relying on16SrRNAand the level of serum TNF-α measured by enzyme linked immunosorbent assay (ELISA).Seventy patients who suffering from CVD (angina, myocardial Infarction and atherosclerosis) aged between 33-86 years have been investigated and compared to twenty of apparently healthy individuals were studies as control group. Twenty six sample (37.14) %detected positive results for Chlamydia pneumonia e by PCR techniques in patient group, while all control group were negative, furthermore current study revealed a highly significant elevation (p<0.01) in the mean level of TNF-α in sera of patients with CVD compared to control group. Also there were considerable differences in the level of TNF-α between Chlamydia pneumonia e positive and negative within the patient group. The present study concludes there is a significant proportion among patients who infected with C. Pneumoniae and these bacteria play an essential role in the pathogenesis of CVD through stimulation of the inflammatory response.
●Ad'hiah,A.H.;Al-Halbosiy,M.M.F. & Al-Jumaily,R.M.Kh (2007). " Modulating the Hematological and cytogenetic Effects of Mitomycin C by Aqueous Extract of Nut Grass(Cyperus rotundus L.).J.Biotechnology Research Center 1(1):43-51.
●Al-Halbosiy,M.M.F.;Al-JumaiJly,R.M.. (2007) ." The Mutagenic Effects of Mitomycin C by Aqueous Modulating Extract of Alhagi graecorum" Ibn Al-Hatham ,J.Pur& 20(2):1-7.
● Al-Halbosiy,M.M.F.; Sulaiman,G.M. & Al-JumaiJly,R.M.Kh. (2008)."The modulator effect of Iraqi propolis Extract on Mitomycin C Induced micronucleus formation in albino male mice". faculty, , 50(1):77-82.
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●Al-Jumaily,R.M.Kh &; Ad'hiah,A.H. & Al-Halbosiy,M.M.F. (2010)."Evaluation of some haematological and cytogenetic effects of Ammi majus seed aqueous extract in albino male mice". Journal of Biotechnology Research Center, (special edition) Vol.4 No.1
●Hatem Tallima, Mohammad Farhan Al-Halbosiy, Rashika El Ridi(2011). Enzymatic activity and immunolocalization of Schistosoma mansoni and Schistosoma haematobium neutral
sphingomyelinase. Molecular & Biochemical Parasitology (178): 23–28.
●Rashika El Ridi, Hatem Tallima, Mohamed Salah, Marwa Aboueldahab, Omar M. Fahmy,Mohammad Farhan Al-Halbosiy, Soheir S. Mahmoud (2012). Efficacy and mechanism of action of arachidonic acid in the treatment of hamsters infected with Schistosoma mansoni or Schistosoma haematobium. International Journal of Antimicrobial Agents (39): 232– 239.
● Mohammed M. F. Al-Halbosiy, Rakad M. Kh. Al- Jumaily, Fadhel M. Lafta & Hussam M. Hassan (2013). Inhibitory Effect of Camel Urine on Neoplastic and Transformed Cell Lines. International Journal for Sciences and Technology Vol. 8, No.2, 31-35.
●Rakad M. Kh. Al- Jumaily, Laith M.J. Al-Shamma, Mohammad M.F. Al-Halbosiy (2013). The ability of rapeseed (brassica napus I.) Seeds oil in inhibiting cell line growth in vitro. Iraqi Journal of Biotechnology, , No.1, 106-113.
● Rakad M. Kh. Al- Jumaily, Nawal MJ Al- Shamma, Mohammad M. Al-Halbosiy ,Laith M.J. Al-Shamma (2014). Anticancer Activity of Sunflower (Helianthus annuns L.) Seeds oil against cell line. Iraqi Journal of Science Vol. 54, supplement No.4, 1003-1009.
●Zainab Y. Mohammed, Saad M. Nada, Mohammad M. Al-Halbosiy, Shaimaa Y.Abdulfattah , Baraa Abdul-Hameed (2014). Cytotoxic effects of Ammi visnaga volatile oil on some cancer cell lines. Journal of Biotechnology Research Center (special edition). Vol.8 No.1, 5-7.
●Mohammad M. F. Al-Halbosiy, Farah Th. Abdulla, Shimaa Y. Abdulfattah, Sabah M. Hussein , Baraa A. Abdul Hameed , Wathiq Q. Shaker and Fathel A.Hasan (2014).Cytogenetic and hormonal study for sample of men infertile in Iraq. International Journal for Sciences and Technology, Vol. 9, 62-65.
● Abeer M. Badr, Mohammed M. F. Al-Halbosiy, Rashika El Ridi. (2015). Differential immune responses toexcretory─secretoryantigens of lung-stage larvae of Schistosoma mansoniin mice and rats. The Journal of Basic & Applied Zoology. 69, 26–33.
● Mohammed Adel Nouri, Mohammed M. F. Al-Halbosiy, Batol Imran Dheeb, Abdulkareem Jasim Hashim. (2015). Cytotoxicity and Genotoxicity of Gliotoxin on human lymphocytes in vitro. Journal of King Saud University – Science.
● Mohammad M. F. Al-Halbosiy, Rakad M. Kh. Al-Jumaily, Fadhel M . Lafta. (2015). The Exposure Effect of Water Pipe Smoke (WPS) on the Total Count leukocyte, Mitotic Index, Micronucleus Formation and Chromosome Aberration in Albino Male Mice. Journal of Biotechnology Research Center, Vol. 9, 89-93.
● Rawaa N. Abdallah1, Raghad K. Allihaibi, Mohammad M.F. Al-Halbosiy and Ali A. Taha (2016). Preparation of Nanoliposomes by Size Exclusion Chromatography to Entrap Soluble Antigens from Leishmania donovani. Iraq Journal of Science (Vol. 57, , : 814-823).
● Niran A.H. Al-Ogaili, Mohammed M.F. Al-Halbosiy, Mahmoud Zayanal Mohammed, Zainab Al-Hawraa Mohammed, Afyaa Diyaa Rushk. (2017). IN VITRO INVESTIGATION OF ANTIBACTERIAL AND ANTILEISHMANIAL ACTIVITIES OF GUM-RESIN OF BOSWELLIA SERRATA USING (0.5%) DMSO SOLUTION AS AN ALTERNATIVE SOLVENT FOR EXTRACTION. World Journal of Pharmacy and Pharmaceutical Sciences, 6(9): 190-199.
● Hujran Abed Al-raheem Abed, Huda F Hasan and Mohammad MF Al-Halbosiy. (2017). Comparative effect between Mesenchymal Stem Cells (MSCs) and Clomiphine citrate on induced premature ovarian failure (POF) in mice. Journal of Entomology and Zoology Studies 2017; 5(5): 955-963. ● Huda F. Hasan, Esraa M. Krair, Mohammad M.F.Al-Halbosiy. (2017). Evaluating anti-anemic effect of Mesenchymal Stem Cells and Oxymetholone on aplastic anemia induced in mice.
● Rajwa Hasen Essa, Mohammad Mahmood F. Al-Halbosiy, Sundus Hameed Ahmed. (2017). Evaluation, Antioxidant, Antimitotic and Anticancer Activity of Iron Nanoparticles Prepared by Using Water Extract of Vitis vinifera L. Leaves. Journal of Advanced Laboratory Research in Biology, 8 (3) :67-73.
●Afnan R. A. Al-Tekreeti, Mohammad M. F. Al-Halbosiy, Batol I. Dheeb, Abdulkareem J. Hashim, Anaam F. H. Al-Zuhairi, Farooq
I. Mohammad. (2018). Molecular identification of clinical Candida isolates by simple and randomly amplified polymorphic DNA-PCR. Arabian Journal for Science and Engineering 43: 163-170.
● Selma M.H. AL-Jawad, Ali A. Taha, Mohammad M.F. Al-Halbosiy, Lamyaa F.A. AL-Barram. (2018). Synthesis and characterization of small-sized gold nanoparticles coated by bovine serum albumin (BSA) for cancer photothermal therapy. Photodiagnosis and Photodynamic Therapy 21: 201–210.
● Huda F. Hasan, Esraa M. Krair, Mohammad M.F.Al-Halbosiy. Evaluating anti-anemic effect of Mesenchymal Stem Cells and Oxymetholone on aplastic anemia induced in mice. (2017).