Association of proprotein convertase subtilisin/kexin type 9 protein and oxidative stress indicators in women with preeclampsia: A case-control study.

Background: Proprotein convertase subtilisin/kexin type 9 (PCSK9) protein is one of the factors associated with oxidative stress and dyslipidemia disorders. Objective: This study aimed to evaluate the lipid profile, PCSK9 levels, and oxidative stress in preeclampsia. Materials and Methods: This case-control study was conducted at Sina hospital in Hamadan University of Medical Sciences, Hamadan, Iran from August 2020-May 2021. The average maternal age of included participants was 30 yr with 30 preeclampsia and 30 healthy pregnant women. After clinical examination, the fasting blood samples were collected, and the serum PCSK9 protein concentration, superoxide dismutase, glutathione peroxidase activities, and glutathione levels were determined by enzyme-linked immunosorbent assay. Total antioxidant capacity, total oxidant status, and malondialdehyde levels were determined manually. Results: The average maternal age of participants were 29.97 ± 4.75 and 31.23 ± 5.85 yr, respectively. The concentrations of total cholesterol, low-density lipoprotein cholesterol (LDL-C), PCSK9, total antioxidant capacity, and malondialdehyde levels were higher in the preeclampsia group compared with control (p < 0.02). Total oxidant status, glutathione levels, superoxide dismutase, glutathione peroxidase activities were lower in the cases group compared with the control group (p < 0.01). The PCSK9 variable had a significant negative association with antioxidant parameters; however, a significant positive association was observed between PCSK9 level and parameters of LDL-C. Conclusion: PCSK9 is associated with increased serum levels of LDL-C and oxidative factors in pregnant women that increase the risk of endothelial damage and hypertension in preeclampsia.

Copper Nanoparticles Induce Apoptosis and Oxidative Stress in SW480 Human Colon Cancer Cell Line.

Cu nanoparticles (CuNPs) have various applications in biomedicine, owing to their unique properties. As the effect of CuNPs on the induction of oxidative stress and apoptosis in the human colorectal cancer cell line SW480 has not yet been studied, we investigated the toxicity and mechanism of action of these NPs in SW480 cells. MTT assay was performed to assess the effect of the particles on the viability of SW480 cells. The levels of oxidative stress were assessed after 24 h of treatment with CuNPs by evaluating the Reactive Oxygen Specious (ROS) production. The antioxidant enzyme activity was assessed using a colorimetric method. To investigate the effect of NPs on cellular apoptosis, Hoechst33258 staining was performed, and the expression of Bax, Bcl-2, and p53 was evaluated by qRT-PCR. The MTT assay results showed that CuNPs inhibited the viability of SW480 cells. Moreover, the increase in ROS production at all three concentrations (31, 68, and 100 µg/ml) was significant. It has been observed that CuNPs lead to increased expression of Bax and p53, and decreased expression of Bcl-2. Hoechst staining was performed to confirm apoptosis. In conclusion, the induction of apoptosis demonstrated the anticancer potential of the CuNPs.

Evaluation of methylenetetrahydrofolate reductase and S-adenosyl-methionine level in male infertility: A case-control study.

Background: Methylenetetrahydrofolate reductase enzyme (MTHFR) plays a key role in regulating folate balance, converting homocysteine to methionine, and producing s-adenosylmethionine (SAM) that plays a role in the methylation process. Objective: This study aimed to determine MTHFR activity and SAM level in men with normozoospermia and oligozoospermia. Materials and Methods: 30 oligozoospermic and 30 normozoospermic men as controls were enrolled in this case-control study. Semen analysis was conducted according to the world health organization criteria. All semen samples were collected after 3-5 days of sexual abstinence. The sperms were evaluated by sperm test video software. All subjects SAM level was measured by enzyme-linked immunosorbent assay kit, and MTHFR were measured manually. Results: 2 groups had a significant difference in sperm morphology (p = 0.02), concentration (p = 0.02) and motility (p = 0.03). The MTHFR activity in normozoospermic and oligozoospermic groups had significantly differences (p = 0.01). The level of SAM in the semen of oligozoospermic men was statistically lower than normozoospermic men (p = 0.03). Also, there was a positive association between MTHFR enzyme activity and SAM level in the normozoospermia group (p = 0.02, ß = 0.67) and oligozoospermia group (p = 0.03, ß = 0.54). Conclusion: MTHFR activity and SAM concentration were statistically lower in oligozoospermia men. It seems they can affect sperm concentration, morphology, and motility.

Beneficial effects of genistein in suppression of proliferation, inhibition of metastasis, and induction of apoptosis in PC3 prostate cancer cells.

OBJECTIVES: Beneficial effects of genistein have been studied in various cancer types but the underlying molecular mechanisms of its actions have not been well established. This study investigated the effects of genistein on caspase-3 and p38 mitogen-activated protein kinase (p38MAPK) as main cellular signalling targets in PC3 prostate cancer cells. METHODS: Caspase-3 and p38MAPK gene expression and intracellular protein levels were determined. Matrix metalloproteinase-2 (MMP2) gelatinase activity and caspase-3 enzyme activity were measured and PC3 cell migration and proliferation potencies were assessed. RESULTS: Genistein induced apoptosis by enhancing the gene expression, intracellular protein level, and enzyme activity of caspase-3. Genistein also inhibited cell proliferation by reducing p38MAPK gene expression and protein level and strongly suppressed metastatic potency of PC3 cells by reducing MMP2 activity. CONCLUSION: Genistein exhibits its beneficial anticancer properties on PC3 cells by reducing metastatic potency and regulating caspase-3 and p38MAPK pathways at different transcriptional and protein levels.

Effects of progesterone nanoparticles on the sperm capacitation and acrosome reaction in asthenozoospermia men.

Low motility is one of the causes of male infertility. In this study, the effects of progesterone solid lipid nanoparticles (SLNs) on sperm capacitation, acrosome reaction, oxidative stress and expression of SPACA1 and MAPK way genes were investigated. Progesterone SLNs were synthesized using the solvent emulsification evaporation method. Twenty asthenozoospermia samples were selected, and sperm and acrosome membrane integrity, acrosome reaction, sperm motility, viability, total antioxidant capacity (TAC), total oxidative status tests and PKA, PTK, P38MAPK and SPACA1 gene expressions were assessed. The synthesized nanoparticles were prepared with the size (187.6 nm), PDI (0.184), EE (85.82%), LP (3.43%) and ZP (-23.5mV). Progesterone SLNs increased sperm and acrosome membrane integrity and TAC (p < .05). Also, the expression of P38MAPK, PKA, PTK, and SPACA1 genes in this group showed a significant increase (p < .001). Progesterone SLNs increased acrosome reaction, sperm capacitation and TAC. Also, it increased the expression of PTK PKA, SPACA1 and P38MAPK genes.

Association of glutamate cystein ligase (GCL) activity Peroxiredoxin 4 (prxR4) and apelin levels in women with preeclampsia.

INTRODUCTION: Preeclampsia is a common disease of pregnancy that is characterized by symptoms such as high blood pressure and proteinuria. Peroxiredoxin 4 (Prx4), is a protein with antioxidant properties which is produced in placenta and protects it from antioxidant stress and recurrent miscarriage. For regeneration of Peroxiredoxin 4 need to glutathione (GSH) and Glutamate-cysteine ligase (GCL) enzyme controls the pathway of glutathione regeneration. Apelin is a paired internal ligand with a G protein coupled receptor and is associated with angiotensin receptor (AT1) as a blood pressure regulator. This study was designed to evaluate GCL enzyme activity and Peroxiredoxin 4, glutathione and apelin levels in serum of women with preeclampsia. MATERIAL AND METHODS: Thirty pregnant women with preeclampsia and 30 healthy pregnant women were enrolled in this study. All participants were diagnosed by clinical examination and confirmation by Obstetrician-Gynecologist. The GCL enzyme activity and concentration of Prx4 and apelin in serum samples were measured using ready-to-use non-competitive ELISA methods while glutathione level was determined using Ellman's reagent. RESULTS: The GCL enzyme activity and Prx4 level were significantly lower in preeclampsia compared with control group (p < 0.05). In addition, marked reductions were observed in the concentrations of glutathione and apelin in preeclampsia compared to the healthy pregnant women (p < 0.05). CONCLUSION: This study identified the role of the GCL and Prx4 system in preeclampsia disorder and may be one of the ways to prevent and reduce the risks of preeclampsia in high-risk women using diet control and stress reduction.

Correlation of thioredoxin reductase (TrxR) and nitric oxide synthase (NOS) activities with serum trace elements in preeclampsia.

BACKGROUND: Preeclampsia is a dangerous disease of pregnancy with symptoms such as high blood pressure and proteinuria. The nitric oxide synthase (NOS) enzyme produces nitric oxide (NO) as a mediator of vasodilation and need to calcium and zinc ions for activity. The thioredoxin reductase (TrxR) as an enzyme containing selenium has been neutralized the damaging effects of oxidants. In this study, our aim was to evaluate the activity of eNOS and TrxR enzymes and the amounts of calcium, zinc, and selenium elements in serum of women with preeclampsia. MATERIAL AND METHODS: Thirty preeclampsia and 30 healthy pregnant women were enrolled in the study after clinical examination and confirmation by Obstetrician-Gynecologist. Venous blood samples were collected and the activity of NOS, TrxR enzymes, and the concentration of zinc, calcium, and selenium elements were measured in serum. RESULTS: The results of NOS and TrxR activities showed significant decreases in preeclampsia compared with control group (p < .05). In addition, concentrations of zinc, calcium, and selenium were markedly lower in preeclampsia compared to the healthy control group (p < .05). CONCLUSION: This study identified the role of the NOS and TrxR activities in preeclampsia disorder and may be one of the ways to prevent and reduce the risks of preeclampsia in high-risk women using diet control and stress reduction.

Evaluation of methylenetetrahydrofolate reductase (MTHFR) activity and the levels of homocysteine and malondialdehyde (MDA) in the serum of women with preeclampsia.

BACKGROUND: Methylenetetrahydrofolate reductase (MTHFR) has a key role in converting homocysteine to methionine so its deficiency might cause homocysteine accumulation and arterial destruction. The aim of this study was to evaluate the activity of methylenetetrahydrofolate reductase (MTHFR), homocysteine, and MDA levels in preeclampsia patients. MATERIAL AND METHODS: In this case-control study, 30 preeclampsia patients and 30 healthy pregnant women as control group were selected and matched for maternal age, gestational age, and BMI. Preeclampsia was confirmed by examining hypertension and urinary proteinuria. Serum homocysteine and MDA level as well as MTHFR activity were measured manually. RESULTS: Homocysteine and MDA levels were significantly different between women with preeclampsia and normal pregnancies (p-value <0.001). Also, the results indicate that MTHFR activity in the two groups was statistically different (p-value <0.001). CONCLUSION: It can be concluded that the regulation of homocysteine and its metabolic enzymes plays a significant role in controlling blood pressure and consequently in the pathophysiology of Preeclampsia and it can be a goal for prevention and treatments.

Association between seminal plasma neopterin and oxidative stress in male infertility: A case-control study.

BACKGROUND: Neopterin is a significant and sensitive marker in estimating the activity of cellular immune system. Oxidative stress plays a role in the etiology of male infertility. Increased reactive oxygen species is accompanied with increase in neopterin level. Hence neopterin may be involved in male infertility. OBJECTIVE: The objective of this case-control study was to determine neopterin level in idiopathic infertile and normospermic men; furthermore, to identify its relationship with oxidative stress markers including total oxidant, malondialdehyde, sperm DNA fragmentation, and total antioxidant capacity of seminal plasma. MATERIALS AND METHODS: Forty seven infertile and forty three normospermic males were selected according to WHO criteria. Their semen and blood samples were taken; subsequently, the levels of neopterin, total oxidant, total antioxidant, malondialdehyde, and sperm DNA fragmentation were measured. RESULTS: The levels of neopterin, total oxidant, and malondialdehyde in seminal plasma of infertile males were significantly higher than those of normospermic group (p=0.038, 0.018, and 0.028, respectively). Furthermore, sperm DNA fragmentation in infertile men was higher than that of control group (p<0.001). Moreover, total antioxidant capacity of seminal plasma in infertile males was significantly lower than that of normospermic subjects (p=0.002). No significant difference was observed in serum neopterin, total oxidant, and malondialdehyde between the infertile and normospermic groups. CONCLUSION: The significant inverse correlation between seminal plasma neopterin and total antioxidant in the infertile males supports a possible role of neopterin in male infertility. Neopterin can be suggested as a marker in monitoring and diagnosis of idiopathic male infertility.

Genistein Induces Apoptosis and Inhibits Proliferation of HT29 Colon Cancer Cells.

Soybean isoflavone genistein has multiple anticancer properties and its pro-apoptotic and anti-proliferative effects have been studied in different cancer cells. However, the mechanisms of action of genistein and its molecular targets on human colon cells have not been fully elucidated. Therefore, caspase-3 and p38 mitogen-activated protein kinase (p38 MAPK) as the main therapeutic targets were investigated in this study at both gene expression and protein levels in HT29 colon cancer cells. The caspase-3 and p38 MAPK gene expression levels were examined by real time PCR whereas flow cytometry technique was performed to determine their intracellular protein levels. The caspase-3 enzyme activity was obtained by colorimetric method while the gelatinase activity of matrix metalloproteinase-2 (MMP2) was determined by zymography. In addition, MTT test, wound healing assay and clonogenic assay were carried out to determine the effect of genistein on HT29 cell viability, migration, and proliferation, respectively. Genistein induced apoptotic death in HT29 cells through activation of caspase-3 pathway at the transcriptional, protein, and enzymatic levels. Moreover, genistein inhibited the proliferation of HT29 cells by reducing of both p38 MAPK gene expression and its active phosphorylated protein level. Also, we showed that genistein strongly suppressed the metastatic potency of HT29 colon cancer cells via the reduction of MMP2 activity. Based on the results of this study, we conclude that genistein may exhibit its anticancer properties on HT29 colon cancer cells by modulating caspase-3 and p38 MAPK pathway at different transcriptional and protein levels.

Contaminated osteochondral plugs: effect of different sterilizing solutions: an experimental study in the rabbit.

BACKGROUND: To determine the efficacy of different antiseptic solutions (Control group (I), Antibiotic solution (II), Chlorhexidine 0.4% (III), and povidone - iodine 10% (IV)) in sterilizing contaminated osteochondral plugs. METHODS: Under sterile conditions, the femoral head and condyles of 20 rabbits were removed and cut into equal osteochondral pieces. A total of 200 osteochondral specimens were obtained. All 200 specimens were dropped on the operating room floor for fifteen seconds and assigned to one of four experimental groups. Group I samples were cultured after washing with normal saline solution (Control group). In other three groups, prior to culturing process, samples were placed in an antibiotic solution after washing with normal saline (Neomycin & Polymyxin) (group II), Chlorhexidine 0.4% (group III), and povidone - iodine 10% (group IV), respectively. RESULTS: In group I, 25 of 50 specimens had positive cultures. Of 50 specimens of group II, III and IV, no positive cultures were found after 10 days. CONCLUSION: All three agents including antibiotic solution, povidone-iodine 10% and chlorhexidine 0.4% seem effective in sterilizing the contaminated osteochondral samples. According to the literature, povidone-iodine has no negative effect on the cartilage metabolism and seems to be a proper choice of decontaminating solution for osteochondral plugs.To the best of the authors' knowledge, such a study on the contaminated osteochondral specimen has not been previously reported in the literature.

Lumbar disk degeneration disease and aggrecan gene polymorphism in northern Iran.

Aggrecan is the major component of intervertebral disk matrix proteoglycan with multiple functional domains. To understand the role of aggrecan polymorphism in a part of exon 12 encoding the CS1 domain in lumbar disk degeneration disease, we have analyzed genomic DNA from 71 patients with the disease and 108 healthy individuals in northern Iran. The AGC1 alleles were determined by PCR followed by gel electrophoresis. Twelve AGC1 alleles ranging from 18 to 29 repeats were detected in patients and controls. The most frequent AGC1 allele was 27, followed by 28 in patients and controls. The shorter AGC1 alleles (< or =24 repeats) were more frequent in patients than in controls (37 vs. 16%, P < 0.001). The odds ratio for lumbar disk degeneration was 3.28 (95% confidence interval 1.62-6.65) in carriers of the shorter AGC1 alleles. Our data suggest that carrying shorter AGC1 alleles with less than 24 repeats could predispose a subject to lumbar disk degeneration disease in northern Iran.