Hematological and serum biochemical studies in fresh water fish exposed to acute and chronic copper and mercury toxicity

H.A., Sawsan,H.M., Amira,M.B., Mostafa,AM.M., Nashaat The Korean Society of Fish Pathology 2017 한국어병학회지 Vol.30 No.1

A total number of 668 apparently healthy fish were obtained from farm to study the effect of two heavy metals in a form of (Copper sulfate and Mercuric chloride) on some hematological and biochemical parameters of blood. The $LC_{50}$ /96 hr. of Cu and Hg were estimated and fish exposed to $\text\tiny{^1/_2}$ $LC_{50}$ for 7 days and for $1/_{10}$ $LC_{50}$ for 8 weeks from each product separately. Results showed decrease in RBCs count, PCV% and Hb in acute and chronic mercury while a significant increase was shown in acute and chronic copper toxicity, total leucocytic count showed decrease in acute mercury toxicity and increase in the chronic case, while in copper toxicity non-significant decrease in acute and significant decrease in chronic toxicity was noticed. Elevated serum urea and creatinine in both acute and chronic mercury and copper toxicity was detected. No changes in total bilirubin in the acute mercury and chronic copper toxicity while significant increase in chronic mercury and acute copper. Elevation of serum AST and ALT in some days of acute toxicity of mercury and copper while in chronic mercury toxicity a significant elevation of both serums AST and ALT were detected .while in chronic copper toxicity serum AST was fluctuated and ALT showed no significant changes. CK study revealed significant decrease in acute mercury with fluctuation in the chronic toxicity while in copper toxicity it showed fluctuation in acute and significant decrease in chronic toxicity. Glucose value decreased in acute and chronic mercury toxicity while in copper toxicity it showed significant increase in the acute and increase followed by significant decrease in the chronic copper toxicity.

Hematological and serum biochemical studies in fresh water fish exposed to acute and chronic copper and mercury toxicity

( Sawsan H. A. ),( Amira H. M. ),( Mostafa M. B. ),( Nashaat Am. M. ) 한국어병학회 2017 한국어병학회지 Vol.30 No.1

A total number of 668 apparently healthy fish were obtained from farm to study the effect of two heavy metals in a form of (Copper sulfate and Mercuric chloride) on some hematological and biochemical parameters of blood. The LC ₅₀ /96 hr. of Cu and Hg were estimated and fish exposed to ½ LC₅₀ for 7 days and for 1/₁₀ LC₅₀ for 8 weeks from each product separately. Results showed decrease in RBCs count, PCV% and Hb in acute and chronic mercury while a significant increase was shown in acute and chronic copper toxicity, total leucocytic count showed decrease in acute mercury toxicity and increase in the chronic case, while in copper toxicity non-significant decrease in acute and significant decrease in chronic toxicity was noticed. Elevated serum urea and creatinine in both acute and chronic mercury and copper toxicity was detected. No changes in total bilirubin in the acute mercury and chronic copper toxicity while significant increase in chronic mercury and acute copper. Elevation of serum AST and ALT in some days of acute toxicity of mercury and copper while in chronic mercury toxicity a significant elevation of both serums AST and ALT were detected .while in chronic copper toxicity serum AST was fluctuated and ALT showed no significant changes. CK study revealed significant decrease in acute mercury with fluctuation in the chronic toxicity while in copper toxicity it showed fluctuation in acute and significant decrease in chronic toxicity. Glucose value decreased in acute and chronic mercury toxicity while in copper toxicity it showed significant increase in the acute and increase followed by significant decrease in the chronic copper toxicity.

Histopathology and residues in fresh water fish exposed to acute and chronic copper and mercury toxicity

Sawsan, H.A.,Amira, H.M.,Mostafa, M.B.,Nashaat, AM.M. The Korean Society of Fish Pathology 2017 한국어병학회지 Vol.30 No.2

A total number of 668 apparently healthy fish were obtained from farm to study the effect of two heavy metals (Copper and Mercury) on histopathology of liver, kidney, spleen, gills and muscles also residues in muscles. The $LC_{50}$/96 hr. of Cu and Hg were estimated and fish exposed to 1/2 $LC_{50}$ for 7 days and for 1/10 $LC_{50}$ for 8 weeks from each product separately. Histopathological findings in acute and chronic mercuric chloride toxicity revealed degeneration and necrosis in the glomeruli, interstitium tissue and epithelium lining renal tubules. The tubular epithelium became necrotic at several places. Eosinophilic hyaline droplets is exist in the cytoplasm of the necrosed cells. Degenerative changes and hyperactivity in melanomachrophage center was seen in the spleen together with some necrotic areas. Necrosis and aggregation of melanomachrophage were seen in the hepatic cells, Hepatic cells showed vacuolar degeneration in the hepatic cells. Gills showed loss in the lamellae of the filaments associated with edema, inflammatory cells infiltration and haemorrhages in the arch. The sarcoplasm of the bundles of the skeletal muscle showed granular degeneration and focal inflammatory cells infiltration between the hyalinized bundles. Mercury residues obtained from these studies in the acute toxicity were 0.22 ppm/gm in the 2nd day, 0.411 ppm/gm in the $5^{th}$ day ended with 0.96 ppm/gm in the $7^{th}$ day. In chronic toxicity it was 1.1320, 1.7140, 2.3620 and 3.5640 ppm/gm respectively from the $2^{nd}$ to the $8^{th}$ week of exposure. In acute and chronic copper toxicity, there was degenerative changes in renal tubules. Melanophores aggregation in the wall of the blood vessels of the spleen and depletion of some of the melanophores in the melanomachrophage were seen together with necrosis in some areas. Congested Mvs (Micro vessels) and vacuolation of hepatocytes were observed. Some areas of hemorrhage and melanophores vacuolar degeneration in the liver were seen. There was mitosis in some areas with displesia of hepatopancreatic cells and eosinophilic granular cells aggregation. Zymogen granules disappeared and there were dyplastic hepatocytes. Congestion in the blood vessels of the gill filaments, associated with massive number of granular eosinophilic cells infiltration were seen in the base of the filaments. There were sever vacuolization and hyalinization in the skeletal muscle bundles. Detection of residues of copper sulfate revealed increase of the amount of copper measured in ppm/gm comparing to the normal control starting from 0.60 ppm/g in the $2^{nd}$ day, 0.67 ppm/g in the $5^{th}$ day and 0.67 ppm/g in the $7^{th}$ day. Result obtained in chronic copper sulfate toxicity revealed gradual increase of the amount of copper which ranged from 0.18 ppm/g at the $2^{nd}$ week to 0.21 ppm/g in the $8^{th}$ week of exposure.

Liver Cancer Stem Cell Induction from Induced Pluripotent Stem Cells

( Said M Afify ),( Ghmkin Hassan ),( Hend M Nawara ),( Hager M Mansour ),( Amira Osman ),( Sadia Monzur ),( Hagar Ali Abu Quora ),( Maram H Zahra ),( Akimasa Seno ),( Yoshiaki Iwasaki ),( Masaharu Sen 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: Liver cancer stem cells represent a small fraction of cells in liver cancer tissues so that studying these cells is very hard. Generation of liver cancer stem cells considered as one of the most important issue in cancer biology research. For this reason, we tried to generate liver cancer stem cells from induced pluripotent stem (iPSCs). Methods: First of all, CM was collected from confluent culture of Huh7 cells. Then, mouse iPSCs cells without MEF feeder cells were cultured in the presence of 50% CM for 4 weeks. The medium was changed every day with fresh medium containing 50% of CM. Mouse iPSCs cultured is the complete medium with LIF were used as a control. The survived cells (5x105 cells) were suspended in HBSS and injected into the liver of BALB/ c nude mice. After 25 days malignant tumor was formed in the liver while benign teratoma was formed by the injection of iPSCs. Tumors were then excised and partly fixed in 10% neutral formalin buffer solution for HE staining and immunohistochemical analysis. The rest of tumors were subjected to rt-qPCR anaylsis and primary culture. Results: Immunohistochemical analysis with liver cancer associated markers and cancer stem cell marker showed that malignant liver tumor was developed. These results indicate that the primary cells from the malignant tumor are rich in CSCs. Conclusions: This model will be very important and useful to assess the significant molecular mechanisms necessary to maintain liver cancer stem cells, which will help in defat liver cancer.

Synergistic Activity of Paclitaxel and Sorafenib Against Liver Cancer Stem Cells

( Hend M. Nawara ),( Said M. Afify ),( Ghmkin Hassan ),( Maram H. Zahra ),( Marwa N. Atallah ),( Hager Mansour ),( Hagar A. Abu Quora ),( Amira Osman ),( Hiroki Kakuta ),( Hiroki Hamada ),( Akimasa Se 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: Cancer stem cells (CSCs), also known as tumor-initiating cells (TICs), are suggested to be responsible for drug resistance and cancer recurrence. Current treatments with conventional chemotherapy are not highly efficient against the cancer stem cells (CSCs). The combination of anticancer drugs, of which functions are different from the other, enhances efficiency compared to the mono-therapy because it targets cancer cells in a synergistic or an additive manner. In this study, the effect of paclitaxel and sorafenib on cancer stem cells (CSCs) developed from mouse iPSCs in very low concentration was evaluated. Methods: To investigate the effect of combination therapy, CSCs were exposed to paclitaxel and/or sorafenib at different concentrations of 1, 2 and 4 nM, respectively. Cell viability was assessed with 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT). The same concentrations of the agents were assessed for the effect on the self-renewal potential of CSCs subpopulation by sphere formation ability. Results: As a result, a combination of sorafenib and paclitaxel significantly reduced the resistance while the CSCs exhibited drug resistance against paclitaxel alone. Also, combination of these agents reduces the self-renewal potential of CSCs when compared to single treatment. Simultaneously, combination significantly suppressed not only the colony formation but also the tube formation of the Cancer stem cells. Conclusions: These results suggest the combination therapy of paclitaxel and sorafenib in low doses be an attractive approach to target cancer stem cells in the future.

The Therapeutic Potential of Amniotic Fluid-Derived Stem Cells on Busulfan-Induced Azoospermia in Adult Rats

Ibrahim Heba F.,Safwat Safinaz H.,Zeitoun Teshreen M.,El Mulla Khaled F.,Medwar Amira Y. 한국조직공학과 재생의학회 2021 조직공학과 재생의학 Vol.18 No.2

BACKGROUND: Busulfan is an alkylating chemotherapeutic agent that is routinely prescribed for leukemic patients to induce myelo-ablation. However, it also results in azoospermia and infertility in cancer survivors. This research was constructed to explore the possible therapeutic role of amniotic fluid-derived stem cells (AFSCs) in improving busulfan-induced azoospermia in adult rats. METHODS: Forty two adult male albino rats were randomized into: (1) control group, (2) azoospermia group, (3) spontaneous recovery group, and (4) AFSCs-treated group, in which AFSCs were transplanted through their injection into the testicular efferent ducts. The assessment included a histo-pathological examination of the seminiferous tubules by the light and transmission electron microscopes. Additionally, the confocal laser scanning microscope was used for confirmation of homing of the implanted cells. Moreover, we conducted an immuno-fluorescence study for detection of the proliferating cell nuclear antigen (PCNA) in the spermatogenic cells, epididymal sperm count, and a histo-morphometric study. RESULTS: AFSCs successfully homed over the basement membrane of the injured seminiferous tubules. They greatly attenuated busulfan-induced degenerative and oxidative changes. They also caused a re-expression of PCNA in the germ cells, leading to resumption of spermatogenesis and re-appearance of spermatozoa. CONCLUSION: AFSCs could be a promising treatment modality for male infertility induced by chemotherapy, as they possess prominent regenerative, anti-apoptotic, and anti-inflammatory potentials. BACKGROUND: Busulfan is an alkylating chemotherapeutic agent that is routinely prescribed for leukemic patients to induce myelo-ablation. However, it also results in azoospermia and infertility in cancer survivors. This research was constructed to explore the possible therapeutic role of amniotic fluid-derived stem cells (AFSCs) in improving busulfan-induced azoospermia in adult rats. METHODS: Forty two adult male albino rats were randomized into: (1) control group, (2) azoospermia group, (3) spontaneous recovery group, and (4) AFSCs-treated group, in which AFSCs were transplanted through their injection into the testicular efferent ducts. The assessment included a histo-pathological examination of the seminiferous tubules by the light and transmission electron microscopes. Additionally, the confocal laser scanning microscope was used for confirmation of homing of the implanted cells. Moreover, we conducted an immuno-fluorescence study for detection of the proliferating cell nuclear antigen (PCNA) in the spermatogenic cells, epididymal sperm count, and a histo-morphometric study. RESULTS: AFSCs successfully homed over the basement membrane of the injured seminiferous tubules. They greatly attenuated busulfan-induced degenerative and oxidative changes. They also caused a re-expression of PCNA in the germ cells, leading to resumption of spermatogenesis and re-appearance of spermatozoa. CONCLUSION: AFSCs could be a promising treatment modality for male infertility induced by chemotherapy, as they possess prominent regenerative, anti-apoptotic, and anti-inflammatory potentials.

E1/E2 of Hepatitis C Virus Genotype-4 and Apoptosis

Zekri, Abdel-Rahman N,Sobhy, Esraa,Hussein, Nehal,Ahmed, Ola S,Hussein, Amira,Shoman, Sahar,Soliman, Amira H,El-Din, Hanaa M Alam Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.7

Several studies have addressed the possible role of hepatitis C virus genotype-4 (HCV GT4) in apoptosis. However, this still not fully understood. In the current study a re-constructed clone of E1/E2 polyprotein region of the HCV GT4 was transfected into the Huh7 cell line and a human apoptotic PCR array of 84 genes was used to investigate its possible significance for apoptosis. Out of the 84 genes, only 35 showed significant differential expression, 12 genes being up-regulated and 23 down-regulated. The highest-up regulated genes were APAF1 (apoptotic peptidase-activating factor 1), BID (BH3 interacting domain death agonist) and BCL 10 (B-cell CLL/lymphoma protein 10) with fold regulation of 33.2, 30.1 and 18.9, respectively. The most down-regulated were FAS (TNF receptor super family), TNFRSF10B (tumor necrosis factor receptor super-family member 10b) and FADD (FAS-associated death domain) with fold regulation of -30.2, -27.7 and -14.9, respectively. These results suggest that the E1/E2 proteins may be involved in HCV-induced pathogenesis by modulating apoptosis through the induction of the intrinsic apoptosis pathway and disruption of the BCL2 gene family.