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Vol.8 No.4 – 4:Evaluation of ameliorating role of avocado Persea americana fruit extract against monosodium glutamate-induced toxicity in pregnant female albino rats and their offspring

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By: Eman H Radwan1,2*, Abdelfattah Elbeltagy1, R Ibrahim1, Gh Tabl3 and Noha Nazeh1

1Zoology Department, Faculty of Science, Damanhour University, Egypt

2Member of National Biotechnology Network (ASRT), Egypt

3Zoology Department, Faculty of Science, Tanta University, Egypt

Abstract

Background: Although monosodium glutamate (MSG) is commonly used as a food additive, the application of higher doses or prolonged uses significantly leads to accumulations in living cells and finally produces cellular toxicity. Persea Americana (avocado) has recently gained substantial popularity and is often marketed as a “superfood” because of its unique nutritional composition, antioxidant content, and biochemical profile. Aim: To evaluate the potential ameliorative role of avocado fruit extract against MSG-induced nephrotoxicity in pregnant rats and their offspring. Thirty-two (24 females and 8 males) albino rats were used in this study. After an acclimatization period of two weeks; the animals were mated, and the pregnant rats were randomly divided into four groups; control (G1), avocado (G2): they were supplemented with 50 mg/kg b.w. of avocado fruit extract, MSG (G3): they were given 3g / kg b.w. of MSG, every other day, and MSG &Avocado (G4): they were given an oral dose of MSG alternatively with avocado fruit extract. At the end of weaning, the female rats and their offspring were sacrificed and the blood was collected and the kidneys were excised to evaluate the renal biochemical and histopathological, and immunohistochemical investigations. Results: In MSG-treated mothers’ rats, the renal cortical sections displayed severe histopathological lesions including little renal corpuscles, atrophied glomeruli, and relatively wide Bowman‘s space. However, the offspring displayed mild renal histopathological lesions compared with their mothers. The immunohistochemical results revealed strong PCNA and Bax expression in the renal tissues of MSG-exposed mother rats and their offspring if compared with the control.  Furthermore, the mean percentage value of positively expressed cells for caspase-3 appeared significantly higher in the renal cells of MSG-induced mother’s rats and their offspring if compared with the control. Additionally, the levels of serum antioxidants (SOD&CAT) and potassium ions appeared significantly lowered while the level of MDA, urea, and creatinine appeared significantly higher if compared with the control.  Co-supplementation of avocado fruit extract to MSG-induced mothers rats and their pups successfully alleviated the histopathological, immune-histo-chemical, apoptotic as well as biochemical changes caused by MSG.  Conclusion: Avocado fruit extract has a powerful ameliorative role against MSG-induced renal toxicity in mother rats and their offspring.

Evaluation-of-ameliorating-role-of-avocado-Persea-americana-fruit-extract-against-monosodium-glutamate-induced-toxicity-in-pregnant-female-albeno-rats-and-their-offspring

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Vol.8 No.3 – 7:Detection of DNA damage by SCD and Rate of Apoptosis DNA by Gel Electrophoresis among infertile males

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By: Nehad Nabil Eskarous1, Sobhy El-Sayed Hassab El-Nabi1, Mohamed Ahmed Abd El Salam2, Khaled Geba1, and Sameh Fayek GamalEl Din2

1- Genetic Engineering and Molecular Biology Department of Zoology, Faculty of Science Menoufia University, Egypt

2- Department of Andrology, Sexology & STDs, Faculty of Medicine, Cairo University, Egypt

Abstract:

Background: DNA damage as Fragmentation has adverse effects on fertilization and embryo development, so it is one of the main causes of a male factor for infertility. Several techniques have been mentioned to elevation this damage. In our study, we determine DNA damage in human spermatozoa by sperm chromatin dispersion (SCD) method and Apoptosis of DNA in human spermatozoa by Optical density in gel electrophoresis in male infertility. Objects and Methods: Semen samples were collected from 100 men and were analyzed by standard light microscopic according to the World Organization (5th edition) for diagnostic fertility. Furthermore, Sperm DNA damage was determined by using Halosperm Kit, then assessment apoptosis by optical density in Gel Electrophoresis. Results: The mean value of DNA by SCD method in infertile males increased with a value of 47.95±10.96 %   when compared with the control value of 21.2 ±2.64 % with (p< 0.00001). On the other hand, the mean value of DNA by measurement of Optical density in Gel Electrophoresis in infertile males decreased with a value of 120.27±18.73 when compare with the control value of 144.4±45 with (p =0.833). Conclusion: The assessment of sperm DNA damage by SCD method and other methods for detection of DNA apoptosis by gel electrophoresis addition to routine semen analysis play important role in the diagnosis and management of male infertility. 

Detection-of-DNA-damage-by-SCD-and-Rate-of-Apoptosis-DNA-by-Gel-Electrophoresis-among-infertile-males

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Vol.8 No.2 – 1:Induction of renal cell line apoptosis by hydrogen peroxide

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By: Khalil A. Alhalfawy1, Bahgat A. Elfiky2, Ahmed M. Zahran3 and Mahmoud M. Zayed2

  1.  Molecular Biology Department, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Egypt
  2. Animal Biotechnology Department, Genetic Engineering and Biotechnology Research Institute – University of Sadat City, Egypt
  3. Internal medicine and nephrology Department, Faculty of medicine – Menoufia University, Egypt

Abstract

Background: Hydrogen peroxide is currently the most widely used as an apoptosis inducer due to its broad cytotoxic efficacy against nearly all cell types. cytotoxicity is achieved over a wide range of doses

Objective: To evaluate the cytotoxic effect of hydrogen peroxide on renal cell lines by detecting RIPK1.

 Methods: In this study, we used a Vero cell line treated with H2O2 at concentrations 0.1 mM, 0.2 mM, 0.4 mM, 0.8 mM, and 1.6 mM and examined after 30 min,1hour,2hours,3hours, 4hours and 5 hours. by using MTT assay to detect cytotoxicity to cell line (by detecting cell viability). Spectrophotometrically measure the absorbance at a wavelength of 570 nm. Measure the background absorbance of multi-well plates at 690 nm and subtract from the 450 nm measurement. Sub-lethal dose to renal cell line is one treated with 1.6 mM for 5 hours. groups group 1: renal cell line as control which not treated by H2O2. group 2: Sub-lethal which renal cell line treated by concentration 1.6 mM of H2O2 for 5 hours. Results: H2O2 is cytotoxic to renal cell line by concentration from 0.1 mM to 1.6 mM. RIPK1 gene expressed in renal cell line treated by H2O2. The sublethal dose reached 1.6 mM for 5 hours.  There is a significant difference between the 2 groups by detecting the expression of the RIPK1 gene.

Induction

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Vol.5 No.2 – 9 : Berberine attenuates cancer cell growth via modulating the cell cycle dynamics but not apoptosis in human colorectal HCT-116 3D spheroid model

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By: Ahmed A. Soffar

Division of Molecular Biology, Department of Zoology, Faculty of Science,

Alexandria University, Alexandria, Egypt

Abstract

Colorectal carcinoma is a cosmopolitan type of cancer with poor prognosis, motivating seeking novel strategies to prevent disease development and progression. The poor prognosis is attributed to the severe toxic side effects of the current therapeutic regimes. Hence, novel less toxic treatment strategies are urgently warranted. Berberine is a natural compound with several biological and pharmacological properties, including anti-fungal, anti-diabetic, cardioprotective effects. Some reports showed that berberine inhibits cell growth by inducing cell cycle arrest and promotion of apoptosis in cancer cells. Importantly, the anticancer potential of berberine in colorectal cancer has not been previously investigated. Hence, this work aims to investigate whether berberine possess anticancer properties against colorectal HCT116 cancer cells. The potential effect of berberine on cell cycle regulation and apoptosis will also be deeply investigated. This work was conducted using the more physiological 3D spheroid culture model that mimics better the impact of the tumor microenvironment as well as the cell-cell interaction in the cellular response to therapy. When compared to the previous studies, this work will explain the mode of action of berberine in more physiological conditions that better mimics the in vivo situation. In order to achieve the goal of this work, spheroid growth assay as well as proliferation assay were performed. Spheroid cell suspensions were further investigated using flow cytometry to assess the cell cycle distribution of cells upon berberine application. BrdU immunostaining was performed to elucidate the S-phase fraction of cells. The proliferation potential and the level of apoptosis were also investigated by Ki67 and Annexin V labelling, respectively. The results showed that berberine attenuated tumor spheroid growth and limits the proliferative capacity of HCT116 cells. This could be attributed to the berberine-mediated G1-phase cell cycle delay. The S-phase fraction of cells was significantly decreased upon berberine application. Unexpectedly, berberine did not induce a significant difference in the % of apoptotic cell fraction of cells as compared to the controls. Collectively, these results suggest that berberine possesses an anti-tumor efficacy in 3D culture preparations via modulating the cell cycle progression. Specifically, berberine induces G1-phase cell cycle delay and decreases the S-phase fraction of cells. Thus, it limits the proliferative capacity of cells. Also, berberine did not induce programmed cell death in the HCT116 spheroids.


Berberine attenuates cancer cell growth via modulating the cell cycle dynamics but not apoptosis in human colorectal HCT-116 3D spheroid model-converted

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Vol.2 No.6 -2 : Anticancer activity of Morus nigra on human breast cancer cell line (MCF-7) the role of fresh and dry fruit extracts.

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By : Ahmed A.1, Ali M. 1, El-Kholie E. 1, El-Garawani I.2* and Sherif N. 1

Abstract

Black mulberry (Morus nigra L.) has a long history of medicinal use in folk medicine. In this study, in vitro anticancer properties of fresh and dry black mulberry (BM) ethanolic extracts on breast adenocarcinoma cell line (MCF-7) were investigated. Cytotoxic effect of both extracts was investigated by neutral red uptake method and the IC50 was assessed. The morphological changes, apoptosis and/or necrosis, were detected by Giemsa staining and acridine orange/ethidium bromide dual fluorescent staining methods, in addition to DNA fragmentation assay by gel electrophoresis and comet assay. Mitotic index was evaluated in MCF-7 groups to investigate the antiproliferative potential of fresh and dry BM extracts. Furthermore, micronucleus and other nuclear abnormalities were also investigated. BM extracts administration gave significant (p < 0.05) morphological evidence of apoptosis and increased cell death after 48 hours more than that of 24 hours-treated MCF-7 cells among fresh and dry extract groups. DNA analysis after 24 h of BM extracts treated groups has no fragmentation while apoptosis is generated after 48h showing more fragmentation in fresh BM groups. The mitotic index records were significantly decreased in dose and time dependent manner showing the better antiproliferative effect with fresh extract treatments. DNA single strand breaks were also increased among the treatment groups at dose and time dependent manner with the best results with fresh extract. It can be concluded that black mulberry fruit is a considerable source of health dietary supplement and its integration for pharmaceutical industries has to be in concern.

2. Anticancer activity of Morus nigra on human breast cancer cell line (MCF-7) the role of fresh and dry fruit extracts.
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Vol.2 No.3 -3 : Adverse effects of monosodium glutamate on the reproductive organs of adult Female albino rats and the possible ameliorated role of carob (Ceratonia Siliqua).

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By : Abd El-Fattah B. M. El-Beltagy1, Heba Attef Elghaweet2

Abstract

Monosodium glutamate (MSG) is a sodium salt of glutamic acid that enhances the flavor of certain foods. It plays an important role in the pathogenesis of anovulatory infertility. This study was carried out to evaluate the histological, histochemical as well as immunohistochemical effects of MSG on the ovaries, uterus and vagina of adult female Wistar rats and the possible ameliorative role of carob (Ceratonia Siliqua). In the present work, twenty- four Wistar rats were divided into four groups (n=6): Control group received 0.5ml saline solution, carob-group, feeding on diet containing 5% carob powder, MSG- treated group received 400 mg/kg b.wt and ameliorative group received MSG 400mg/kg b.wt simultaneously with feeding on diet containing 5% carob powder. In MSG treated rats, the serum estrogen level was significantly decreased. The histopathological results showed ovarian atretic follicles, fragmented oocyte, vascular congestion, and vacuolated stroma. The uterus exhibited a comparative reduction in the endometrial thickness and loss of endometrial glands associated with cellular hyperplasia and reduced vacuolated cells. In MSG treated group, the vaginal epithelium appeared relatively thin, folded and hypertrophied with sign of cornification. Histochemically, a weak reaction for PAS stain was remarked in investigated sections of MSG- treated rats. On the other side, an intense positive reaction for silver stain was recorded in the ovarian stroma and peripheral layers of atretic follicles as well as spreading in endometrium of uterus and lumina sheath of vagina. Immunohistochemically, MSG-treated group exhibited intense positive reaction for CD31and CD68 and activated caspase3 compared to carob-ameliorated MSG and the control. The authors concluded that, feeding of MSG-treated group on diet containing 5% carob powder led to marked amelioration of histopathological lesions induced in ovaries, uterus and vagina by MSG attributed to the biological benefits of its phytochemical components.

3. Adverse effects of monosodium glutamate on the reproductive organs of adult Female albino rats and the possible ameliorated role of carob (Ceratonia Siliqua).
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