Jentashapir Journal of Cellular and Molecular Biology
Volume:11 Issue: 3, Sep 2020

While radiotherapy is the important modality in the treatment of breast cancer cells, radioresistance of some tumor cell lines such as MDA-MB-231 is still a limitation that must be considered.

The present study was done to examine the effect of the conditioned medium of the human umbilical cord Wharton's jelly stem cells (hWJSCs + CM) on the radiosensitivity of MDA-MB-231 cells in combination with megavoltage-radiations.

Groups are Control, CM, GY, and GY + CM. In irradiation groups, breast cancer cells were exposured with 4, 6, and 8 Gy radiation. Each group includes different doses of the conditioned medium of hWJSCs (25%, 50%, and 75%).

The MTT assay showed that the proliferative activity of Gy + CM groups at all doses of condition medium decreased significantly compared with the control, rather than Gy groups. Trypan blue viability test showed that the survival rate of MDA-MB-231 cells significantly reduced in the CM and 8 Gy + CM groups compared with the control group, rather than Gy groups. MDA-MB-231 cells lost their normal spindle shape and became thinner and longer after 48h of treatment and the number of cells sharply reduced in Gy + CM groups compared with the control group, rather than Gy groups. These changes were accompanied by inducing significant up-regulation of Interleukin-6 (IL-6) in the 4 Gy + CM and 8 Gy + CM groups compared with the control group, rather than Gy groups and as a consequence, a decrease in the amount of transmembrane tumor necrosis factor-α (tmTNF-α) as a pro-inflammatory cytokine in the Gy + CM groups compared with the control group, rather than Gy groups. Also, we indicated that the radiosensitivity of breast cancer cells was probably enhanced by an increase in different doses of the conditioned medium of stem cells.

Treatment of the MDA-MB-231 cells with hWJSCs + CM plus radiotherapy inhibited the growth and proliferation of cancer cells and this method is a novel strategy for breast cancer therapy by overcoming radioresistance.

Aminoglycoside antibiotics such as gentamicin are used to cure bacterial infections in humans and other animals, but they can cause nephritic damage, as well. Nephrotoxicity is one of the side effects of gentamicin.

The objective of this study was to investigate the effects of toxicity induced by gentamicin on the kidney of killifish Aphaniops hormuzensis. Also, we aimed to study the expression pattern of Wt1 and MMP9 genes by real-time PCR in response to this toxicity.

First, 10 µg/g (sub-lethal dose) gentamicin was given to adult fish. The kidney tissues were dissected and preserved in 10% formalin for a 24-hour; then, they underwent standard histological procedures. The sections were prepared at 3 μm and stained with Haematoxylin & Eosin (H&E). The slide microphotography process was done by an Olympus CH2 microscope. The RNA was isolated, and cDNA was synthesized with a standard protocol, and the expression patterns of Wt1 and MMP9 genes were examined by real-time PCR.

Nephrotoxicity occurred 10 hours after the injection of gentamicin, and the injury was detected in the epithelium of kidney tubules. The kidney tubule regenerated itself within 10 days post-injection (dpi). On 7 dpi, the nephrogenic body formation occurred and was differentiated into renal nephrons. The Wt1 gene was upregulated (two-fold) on 5 dpi after kidney damage and then had a down-regulation on 7 dpi when the kidney began to regenerate. The MMP9 gene showed increased expression in comparison with the control sample in the study days, and this expression increased on 7 dpi by 6.6 folds.

The results of this study, for the first time, highlighted that nephritic damage appears in the kidney of A. hormuzensis after toxicity induced by gentamicin and that changes in the expression of the examined genes are consistent with their roles in the process of renal regeneration in this species.

Liver fibrosis is a reversible response to wound-healing that occurs in most forms of chronic liver damage, beginning with the activation of hepatic stellate cells (HSCs). The increased expression of genes, such as beta-converting growth factor (TGF-β) and actin-alpha smooth muscle (α-SMA) indicates the activation of HSCs. During liver damage, HSCs are activated and converted to myofibroblasts. As a result, the expression of TGF-β and αSMA genes in HSCs increases and leads to liver fibrosis. High fructose intake is known to have harmful effects on human health. Due to the persistent increase in high fructose intake via many beverages and foods in industrialized countries, much concern has been raised about the effect of fructose on liver damage, but its role in activating human HSCs has not been studied.

We aimed to investigate the effect of high fructose concentration on human HSCs activation by measuring the level of mRNA expression of TGF-β and α-SMA genes involved in liver fibrosis.

Human HSCs were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) plus 10% Fetal Bovine Serum (FBS) at 37°C in 5% CO2. Cells were incubated in media containing 25 and 30 mM fructose for 48 h. The control group was incubated in DMEM without fructose. The cells were serum-starved for 24 h before treatment. Then, the total RNA was extracted, reversely transcribed into cDNA, and underwent Quantitative Real-time PCR (qRT-PCR).

The results indicated that the mRNA expression of TGF-β and αSMA genes significantly increased by treating with 25 and 30 mM fructose in HSCs when compared to the control group (P < 0.05).

The increase in the mRNA of TGF-β and αSMA genes is used as a standard marker for HSC activation, leading to liver fibrosis. The results demonstrated that high fructose concentration could activate HSCs and increase the levels of TGF-β and αSMA in these cells. Thus, controlling fructose consumption and identifying the mechanism of fructose action is important to treat and reduce liver injury.

Cancer is one of the most complicated diseases with various treatments, which each has its special side effects. So, pharmaceutical companies are intended to develop new drugs with minimum side effects.

The current study aimed to investigate the cytotoxicity effects and the redox potential of alcoholic extract of Oleaster leaf on liver carcinoma cell line (HepG2).

Oleaster leaves were collected from Qazvin (Iran), and the alcoholic extract of the plant leaves was prepared. HepG2 cells were cultured in DMEM medium and treated with 50, 100, 200, 400, and 600 μg/mL of the extract. The cytotoxicity effect of the extract was evaluated using the MTT and the Neutral Red assays. Redox potential in HepG2 cells was assessed using NO, catalase, and GSH tests. The expression of Bax and bcl-2 genes in HepG2 cells was evaluated for apoptosis analysis.

The results showed that the extract could significantly (P < 0.001) reduce the viability of HepG2 cells. Also, the extract significantly increased the amount of released NO, catalase activity, and GSH concentration. RT-PCR results showed that Oleaster leaf extract significantly change the expression of bax and bcl-2.

The results showed that the leaves of the Oleaster plant contain compounds with cytotoxicity properties, so it can be considered as a potent candidate for liver cancer treatment.

Congenital central hypothyroidism (CCH) is a rare autosomal recessive disease caused by mutations in the thyroid-stimulating hormone β subunit (TSHβ) gene. Since patients with CCH do not experience increased serum levels of TSH, the diagnosis is usually delayed, which leads to negative consequences in the neonatal TSH screening. Genetic diagnostic studies enable us to identify affected relatives at high risk for rapid diagnosis and treatment of the disorder.

This study aimed to investigate genetic variations in the TSHβ gene for the first time in Iranian patients with CCH.

Seven children affected by congenital TSH-deficient hypothyroidism were investigated for mutations in TSHβ. Variable TSH levels in these patients ranged from low values for diagnosis to significant values, so central hypothyroidism was assumed due to mutations in the TSHβ gene.

We identified two novel heterozygous (F11Y and G106R) and one homozygous (T14A) missense mutations in the coding sequence of exons 2 and 3. One of the new heterozygous mutations (F11Y) and a homozygous (T14A) missense mutation were found in exon 2 of the TSHß-subunit gene. The novel mutation G106R in exon 3 was found in three pediatric patients with congenital hypothyroidism. c.40A > G (T14A, rs10776792) appears to be the most common genetic variation associated with TSH deficiency. The others were c.32T > A in exon 2 and c.316G > C in exon 3, which resulted in a change from phenylalanine to tyrosine (p.F11Y) and glycine to arginine (G106R), respectively.

The identification of these mutations for the first time in Iranian patients suggests that CCH is more common than previously recognized, and the TSHβ gene may be the mutational hot spot.

Dairy products are an important part of the human diet due to their health benefits. Some dairy and natural products contain probiotic organisms that make these products have anticancer properties. The most important food fermenting microorganisms are lactic acid-producing bacteria, among which the lactobacillus genus is a very prominent microorganism in terms of their ability to reduce the risk of cancer and have anti-proliferative properties.

In this study, the biochemical characteristics, genetic characteristics, and anti-proliferative and inhibitory effects of lactobacilli isolated from “Shoor” traditional dairy products were evaluated.

Appropriate dilutions of the collected Shoor samples from the region of Azerbaijan were made in normal saline and pour plated on MRS agar and incubated at 37ºC. The isolates were identified biochemically and molecularly. MRS broth medium was used to extract the supernatant of isolated strain. These compounds were then used to test their cytotoxicity on HCT116 cancer cells.

Three isolates were isolated from Shoor samples. According to cellular assays, the supernatant of ST1 isolate was determined as the most significant compound with anti-proliferative properties on the cancer cell line (P < 0.05). The biochemical properties of the isolates were also determined. The molecular results showed that the isolate was 99% compatible with Lactobacillus paracasei.

The “Shoor” traditional dairy product has probiotic potential, and the cytotoxic effects of bacterial metabolites are dependent on concentration and time, and with increasing of these parameters, reduction of cell survival in cancer cells is observed. It is very important to study the application of the microorganisms of this probiotic product as the starter.

Staphylococcus aureus (S. aureus) is a major cause of nosocomial infections in humans and animals. Because of the widespread resistance to antibiotics, microbiologists are trying to find other therapeutic interventions such as phage therapy for bacterial infections.

The present study aimed to isolate staphylophages with lytic effects on methicillin-resistant S. aureus (MRSA) clinical isolates as a potential alternative agent to antibiotic therapy.

This experimental, descriptive study is performed in the Microbiology Laboratory of Shahrekord University (Iran) from September 2018 to March 2019. Two cocktails of staphylophages were isolated from Isfahan (Iran) urban sewage samples. The double-layer agar method was used to detect lytic phages. Morphology characteristic by transmission electron microscopy (TEM) images was used to identify staphylophages. One hundred and thirty three S. aureus were isolated from clinical samples of two teaching hospitals in Isfahan and Shiraz, Iran. Methicillin resistance and the presence of the mecA gene were determined by the disk diffusion method and polymerase chain reaction (PCR) assay, respectively. The phage susceptibility of mecA positive isolates was determined by plaque assay.

Two staphylophage cocktails were prepared, which had lytic effects on forty-four MRSA isolates. Cocktails 1 and 2 lysed 19 (14.2%) and 25 (18.7%) isolates, respectively. Of 133 S. aureus isolates, 88.7% carried the mecA gene.

Different bacteriophages in two phage cocktails had relatively good lytic effects on S. aureus clinical isolates. Therefore, phage cocktails may be an appropriate alternative to antibiotics against S. aureus.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel pathogen that has triggered a pneumonia outbreak, and despite the measures, the pandemic still continues to occur.

The molecular docking analysis was used to test whether the human immunodeficiency virus 1 (HIV-1) protease inhibitory peptides. These marine polypeptides were isolated from the hydrolysate of Pacific oyster.

Molecular docking process was performed using Molegro Virtual Docker software. The protein data bank file of the crystal structure of COVID-19 main protease in complex with an inhibitor N3 (ID 6LU7) was obtained from the PubChem data source. After preparing protein and removing water and internal ligand, the major cavity was selected for the next step, the docking procedure. Afterward, the MolDock score, Rerank score, Total interaction energy (between energy), and HBond item were calculated. The Remdesivir was used as a positive control in the docking project.

The results of the docking step were evaluated based on several bioinformatics docking scores, including MolDock score, Rerank score, Total interaction energy (between energy), and HBond. The hydrogen bond of remdesivir was -6.03673, and Leu-Leu-Glu-Tyr-Ser-Ileu polypeptide was -6.44185. The Rerank score of remdesivir was -98.9254 and for Leu-Leu-Glu-Tyr-Ser-Ileu polypeptide was -107.821. Of the two screened Pacific oyster polypeptides, the score of Leu-Leu-Glu-Tyr-Ser-Ileu ligand was higher than remdesivir.

This study demonstrated that Pacific oyster compounds may have the potency to be evolved as an anti-COVID-19 main protease drug to fight against the novel coronavirus; however, preclinical and clinical trials are needed for further experimental and/or clinical scientific validation.

Liver fibrosis is a reversible response to wound-healing that occurs in most forms of chronic liver damage, beginning with the activation of hepatic stellate cells (HSCs). The increased expression of genes, such as beta-converting growth factor (TGF-β) and actin-alpha smooth muscle (α-SMA) indicates the activation of HSCs. During liver damage, HSCs are activated and converted to myofibroblasts. As a result, the expression of TGF-β and αSMA genes in HSCs increases and leads to liver fibrosis. High fructose intake is known to have harmful effects on human health. Due to the persistent increase in high fructose intake via many beverages and foods in industrialized countries, much concern has been raised about the effect of fructose on liver damage, but its role in activating human HSCs has not been studied.

We aimed to investigate the effect of high fructose concentration on human HSCs activation by measuring the level of mRNA expression of TGF-β and α-SMA genes involved in liver fibrosis.

Human HSCs were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) plus 10% Fetal Bovine Serum (FBS) at 37°C in 5% CO2. Cells were incubated in media containing 25 and 30 mM fructose for 48 h. The control group was incubated in DMEM without fructose. The cells were serum-starved for 24 h before treatment. Then, the total RNA was extracted, reversely transcribed into cDNA, and underwent Quantitative Real-time PCR (qRT-PCR).

The results indicated that the mRNA expression of TGF-β and αSMA genes significantly increased by treating with 25 and 30 mM fructose in HSCs when compared to the control group (P < 0.05).

The increase in the mRNA of TGF-β and αSMA genes is used as a standard marker for HSC activation, leading to liver fibrosis. The results demonstrated that high fructose concentration could activate HSCs and increase the levels of TGF-β and αSMA in these cells. Thus, controlling fructose consumption and identifying the mechanism of fructose action is important to treat and reduce liver injury.