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Abstract Antimicrobial peptides, such as nisin, are proposed as promising agents for cancer treatment. While glycation has been recognized as an effective method for enhancing various physicochemical properties of nisin, its anticancer effects remain unexplored. Therefore, we aimed to assess the anticancer potential of glycated nisin against MDA-MB-231 cells. The MDA-MB cells were treated with increasing concentrations of nisin and glycated nisin for 24, 48, and 72 h. The IC50 values for nisin were higher than those for glycated nisin. Glycated nisin at concentrations of 20 and 40 µg/mL decreased cell viability more than nisin at the same concentrations. The rate of apoptosis in the group treated with 20 µg/mL of nisin was lower compared to other treatment groups, and no significant difference in apoptosis rates was observed at different time points (p > 0.05). However, in the glycated nisin groups with concentrations of 10, 20, and 40 µg/mL, the level of apoptosis was very high after 24 h (73–81% of cells undergoing apoptosis). Overall, our study suggests that glycated nisin exhibits stronger cytotoxic effects on MDA-MB-231 cells, primarily involving the induction of apoptosis. This indicates its potential utilization as an alternative approach to address the issue of drug resistance in cancer cells.

The present study aimed to evaluate the acute phase proteins (serum amyloid A, alpha-1 acid glycoprotein, and C-reactive protein) profile in canine and feline fetal fluids during the second half of gestation. Seventeen pregnant domestic short-hair queens and 19 pregnant mixed-breed bitches were enrolled. Animals were divided into three groups according to their gestational age (30–39, 40–49, and 50–60 days). After ovariohysterectomy and sample collection, the concentration of the mentioned APPs was measured by the ELISA method. There was an increasing trend in the concentration of APPs in canine fetal fluids alongside increasing pregnancy age, but these changes were not statistically significant. Canine amniotic SAA concentration was significantly higher than allantoic. The highest concentration of SAA and CRP in the feline fetal fluids was observed at days 40–49, and the maximum concentration of AGP was observed at days 50–60. For the first time, the current study investigated the APPs profile in canine and feline fetal fluids during pregnancy. The maximum concentration of APPs was observed at the end of canine pregnancy and at days 40–49 of feline pregnancy.

Abstract Background The aim of current study was to construct, express, purify and immunogenicity evaluate of a novel recombinant fusion protein including Pyruvate dehydrogenase beta subunit (PDHB) and high antigenic region of lipoprotein P80 of Mycoplasma agalactiae. Using bioinformatics tools, antigenicity and physiochemical properties of fused protein were assessed. Material and methods The recombinant fusion protein of GST-PDHB-P80 were expressed in pGEX4T-1 and purified then verified by Western blot assay. The purified protein was successfully used for immunization of mice. 30 female BALB/c mice were divided into three groups (10 mice per each group) injected with GST-PDHB-P80, inactivated bacteria vaccine and PBS as negative control, separately. Results Western blot analysis confirmed the interaction between the immunized mice serum and the blotted recombinant protein GST-PDHB-P80, demonstrating the immunogenicity of this protein. Moreover, the sera of vaccinated mice with inactivated bacteria vaccine, containing whole cell proteins, detected the recombinant protein GST-PDHB-P80 confirming the antigenicity of PDHB-P80. Negative control displayed no reactivity with GST-PDHB-P80. Conclusion We proposed a novel designed chimeric protein of Mycoplasma agalactiae as a potential marker for serodiagnostic assays but still further field research is required.

Abstract Background The current study was aimed at evaluating the role of the N-SH2 domain of SHP-2 as a partner protein in the expression of a toxic peptide, laterosporulin (LTS). We also investigated its effects on the formation of the disulfide bond and functional folding of the peptide in vitro. The N-SH2-LTS protein was expressed as a His-tagged fusion protein, capable of undergoing enzymatic cleavage. Results Based on the data presented herein, the total yield of the folded fusion protein from inclusion bodies was found to be about 105 mg/l, demonstrating a high-level of heterologous expression. After enzymatic cleavage, 1.5 mg of the folded recombinant laterosporulin was obtained from each 10 mg of the fusion protein. The purity of the recombinant laterosporulin was analyzed by RP-HPLC, to yield peptides with suitable purity (85%). Conclusions Our findings indicated the advantages of using the N-SH2 domain of SHP-2 as a rapid and easy approach not only in producing easy target proteins but also in its function as a chaperone. N-SH2 domain of SHP-2 can influence on the purification of laterosporulin at reasonable yield and in a cost-effective fashion. The N-SH2 domain of SHP-2 as a protein chaperone may be potentially favorable to produce other proteins with disulfide bonds.

Background/Aims Ulcerative colitis (UC), along with Crohn’s disease, is one of the main types of inflammatory bowel disease (IBD). On the other hand, deregulated autophagy is involved in many chronic diseases, including IBD. In this study, we aimed to investigate the role of Atg5 and microRNA-181a (miR-181a) in the pathophysiology of UC. Methods Colon biopsy, stool, and blood samples of 6 men and 9 women were confirmed for UC. Also, 13 men and 17 women were selected as healthy control (HC). Enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to measure the Atg-5 content of the colon biopsies. Besides, the serum and stool levels of Atg5 were measured using ELISA. Moreover, the total RNA of blood cells was extracted and evaluated for the expression of miR-181a. Results We found 1.2 ng/mL versus 0.46 ng/mL, 0.34 ng/mL versus 0.24 ng/mL, and 0.082 ng/mL versus 0.062 ng/mL of Atg5 in stool, intestinal tissue, and serum of UC and HCs, respectively. There was no significant difference in the expression of miR-181a in the blood samples of UC and HCs. Immunohistochemistry showed high positivity without any significant difference between the 2 groups in the quantitative analysis. Conclusions The significant difference observed between the stool Atg5 content of the HCs and UC patients may provide new insight into using this protein as a diagnostic biomarker, however, considering the small size of our studied population further studies are needed.

Background: Cancer is one of the major health problems worldwide and natural resources are being explored to develop anticancer drugs with fewer side effects. Iranian propolis contains components including flavonoids and polyphenols and has various medicinal properties. The aim of this study was to investigate the effect of Ethanolic Extract of Sirch Propolis (EESP) on three breast cancer cell lines. Methods: The MDA-MB-231, SKBR-3 and MCF-7 cells were treated for 24 and 48 h at the presence of 1% and 10% fetal bovine serum (FBS) concentration. MTT, BrdU and flow cytometry assays were used for measuring cytotoxicity, cell proliferation and apoptosis. Results: The highest cytotoxicity was seen on MDA-MB-231 cell at the presence of 1% and 10% FBS respectively following 48 h treatment. BrdU assay showed that treatment with 200 μg/ mL of EESP at the presence of 1% FBS for 48 h, reduced proliferation of MDA-MB-231 cell to 75% and that of MCF-7 and SKBR-3 cells to 70% and 60% respectively. Cell cycle analysis by flow cytometry showed that EESP at 200 μg/mL for 48h, induced G0/G1 phase arrest in MCF-7 and SKBR-3 cells and G2/M, S phase arrest in MDA-MB-231 cell. The cytotoxic effects of EESP were primarily found to be due to the induction of early stage apoptosis on SKBR-3 cell and early and late stage apoptosis on MCF-7 and MDA-MB-231 cells. Conclusion:The results demonstrated that EESP is a natural anticancer mixture capable of reducing breast cancer cells proliferation and inducing cell cycle arrest and apoptosis in them.

Abstract Type 2 diabetes mellitus (T2DM) is a heritable metabolic perturbation, rapidly growing across the world. Primary recognition of susceptible individuals with a family history of type 2 diabetes (FHD) in the prediabetes stage could delay the onset of T2DM or reduce complications induced by diabetes. This study aims to evaluate the expression levels of miR‐21, miR‐126 as noninvasive predictive biomarkers in individuals with genetic predisposition and investigate the correlation of miRNAs and cardiometabolic risk factors. Our study demonstrated that miR‐21 expression has a notable elevate in both groups of T2DM and pre‐T2DM. miR‐21 expression was distinguished in the pre‐T2DM and T2DM from the nondiabetic individuals by ROC curve analysis with AUC of 0.77 (95% CI 0.65–0.90; p = 0.0004) and AUC of 0.78 (95% CI 0.64–0.92; p = 0.0042), respectively. The relative gene expression of miR‐126 was nearly equal among groups. miR‐21 expression was positively associated with glycosylated hemoglobin (HbA1c), fasting blood sugar (FBS), and triglyceride (TG) and might have diagnostic value for T2DM and pre‐T2DM. This study has revealed that the expression level of miR‐21 can be considered as a non‐invasive and rapid tool for distinguishing pre‐T2DM and T2DM counterparts from healthy individuals.

Abstract Purpose Ulcerative colitis (UC) is a lifelong disease with unclear etiology and increasing prevalence worldwide. Autophagy has been reported to play roles in the pathogenesis and progression of UC. Here, we aimed to analyze the expression of autophagy related protein 10 (ATG10) and its regulator, micro‐RNA (miR) 519a, in UC patients. Methods The level of ATG10 in the serum, stool, and colon biopsies from 15 UC patients and 30 non‐UC healthy individuals (HC) group was measured by ELISA. Also, the blood level of miR‐519a was investigated by quantitative real‐time PCR. Results We found 13.63 ng/ml versus 0.99 ng/ml, 11.01 ng/ml versus 1.11 ng/ml and 6.41 ng/ml versus 3.21 ng/ml of ATG10 in the stool, colon tissue, and serum of UC and HC, respectively. There was no significant difference in the expression of miR‐519a in the blood samples of UC and HC. Conclusions ATG10 might be a potential non‐invasive diagnostic biomarker for UC.

Abstract Background Quercetin (QT) as a bioactive flavonoid has a potential therapeutic activity for numerous neuronal injuries and neurodegenerative diseases. However, the low absorption rate of QT, especially through the blood-brain barrier, restricts its bioactivity in the body. The current research took the advantage of superparamagnetic iron oxide nanoparticles (SPIONs) to enhance the bioavailability of quercetin. Methods Quercetin conjugated with SPIONs was prepared by means of nanoprecipitation method and was characterized by X-ray diffractometer, scanning electron microscope, and Fourier transformed infrared spectrometer analyses. Wistar male rats were orally fed by gavage with QT and QT-SPION at 50 and 100 mg/kg daily doses for 7 days. Using high-performance liquid chromatography (HPLC) method, biodistribution of QT was evaluated in plasma and brain tissue. Results The outcomes of this research revealed a higher concentration in the plasma and brain of the rats fed with QT-SPION in comparison to free QT. Conclusion The results of this study confirm that SPION as a targeted drug delivery system enhances the bioavailability of quercetin in the brain about ten folds higher than free quercetin and could be used for the treatment of neurodegenerative disorders.

In this study, we attempt to find out whether chronic low dose 3-Iodothyronamine (an endogenous metabolite of thyroid hormone) administration could modulate liver de novo cholesterol synthesis, the same as thyroid hormones. Eighteen male mice were divided randomly into treatment (n=10) and control (n=8) groups. The experimental procedure was applied for 7 days during which test group received T1AM whereas the control group received dimethyl sulfoxide and normal saline. The liver was analyzed for HMG-CoA reductase concentration and hepatic lipase activity whiles cholesterol, LDL and HDL concentrations were measured in the blood serum. There was non-significant decrease in HMG-CoA reductase concentration (224±21.2 versus 187±32.5) in test group compared to control. Interestingly LDL and cholesterol concentrations exhibited significant decrease in test group versus the control. There was non-significant decrease in hepatic lipase activity (771±316 versus 645±317) in test group versus the control. It appears that T1AM reduced serum LDL and cholesterol just like T3, in contrast, it decreased liver cholesterol biosynthesis contrary to THs.

Abstract Background: Silver nanoparticles are capable of inducing toxicity in living organisms. Silver nanoparticles can induce some effect in the liver. Thus silver nanoparticles, due to their wide spread effects, can also affect on hepatic, hematological, and oxidative stress factors. Ginger because of its powerful antioxidantal compounds can influence the toxicity effects of silver nanoparticles in different parts of the body. The aim of this study was to investigate the protective effect of hydroalchoholic extract of ginger on cytotoxic silver nanoparticles on enzymes, hematological parameters, blood oxidative stress markers, and hepatic apoptosis in Balb-c mice. Materials and Methods: In this study, 48 rats of Balb-c race Syrians were selected and devided into 4 groups, each consisting of 12. They were treated for a period of 35 days; the first group (control) received distilled water, the second group received nano silver, the third group received ginger extract, and the fourth group received both nanosilver and ginger extract at the same time. Bleeding was done to measure hematological factors, liver enzymes, and oxidative stress; then liver tissue was removed for evaluation of apoptosis. Data were compared using SPSS software and one-way ANOVA. Results: Enzymes AST , ALT , ALP, GGT and LDH as liver factors showed significant differences in the groups of the study. Hematological factors including of WBC , RBC , Hb , HCT , MCV , MCH , Plt , Lymphocyte and Monocyte showed significant differences in all the groups. Of oxidative stress factors , only GPX showed significant difference between groups, while no significant difference was observed in other oxidative stress parameters in the blood. Changes in apoptosis showed significant differences in all groups of the study. Conclusion Based on the findings the study, silver nanoparticles with their side effects in different parts of the body can induce changes in various factors and enzymes. Ginger can compensate ,and modify to some extent these side effects. Such effectiveness of ginger can probably be due to its special ingredients.

Some hormonal disturbances have been demonstrated in starvation, but in ruminants such as goats, the role of galanin in adaptation to starvation or endocrine functions is not well studied. The present study was conducted to assess the effect of long term starvation on galanin, leptin, thyroid hormones, insulin, prolactin, growth hormone, ghrelin and factors involved in energy metabolism including HDL, Cholesterol, β-hydroxybutyrate, glucose, NEFA, TG and VLDL concentrations in adult goats. Eight non-lactating non-pregnant goats aged 4-5 years and BCS 3 were randomly divided to control and test groups. The animals were trained to eat their daily forage ration during a 10 day period. The experimental procedure was applied for 20 days, during which control group received 120% of maintenance energy, while the test group was supplied with 80% of maintenance energy for the first 10 days and with 40% of maintenance energy for another 10 days. Blood samples were collected at day 10 of training and 2, 4, 10, 12, 14 and 20 days after beginning of starvation. Blood parameters were measured according to standard procedures. No significant difference was observed in the concentrations of cholesterol, fT3, T4, T3, growth hormone, NEFA, insulin and ghrelin between control and test groups (P=0.05). There was significant difference in galanin, leptin, fT4, HDL, glucose, TG, VLDL and prolactin concentrations between control and test groups (P=0.05). Control of energy balance and the role of galanin in adaptation to long starvation or endocrine functions in goat are different from other species.

Objective(s):Psoriasis is an autoimmune disease that appears on the skin. Although psoriasis is clinically and histologically well characterized, its pathogenesis is unknown in detail. The aims of this study were to evaluate the proteome of psoriatic patients' sera and to compare them with those of normal healthy human to find valuable biomarkers. Materials and Methods: In a case-control study, twenty cases of white patients with psoriasis vulgaris, 10 males and 10 females and sixteen healthy controls, 8 males and 8 females were enrolled in the study. The serum protein expression patterns obtained after depletion of albumin were compared by using two dimensional gel electrophoresis (2-DE) coupled to MALDI/TOF-TOF to identify disease associated proteins. Results: Differential expression of nine protein spots representing four unique proteins including alpha-1 antitrypsin, retinol binding protein, keratin 10 and an unknown protein (with pI 6.47 and molecular weight of 19941 Da), between psoriatic and healthy human serum were found. Furthermore, expression of four new alpha-1 antitrypsin isoforms with different molecular weight and isoelectric point were observed in psoriatic serums in this research for the first time. Conclusion: A unique proteomic profiling with abnormal expression of alpha-1 antitrypsin and presence of keratin 10 in sera of psoriasis patients were observed that may constitute new and useful findings of psoriasis and offer a clue to a better understanding of the inflammatory pathway.

Objective: Scutellaria luteo-caerulea Bornm. & Snit. is one of the species of genus Scutellaria, within the family of the Lamiaceae, that is used for immune system stimulation and antibacterial effects in traditional medicine in Iran. The aims of this study were to analyze essential oils and mineral element contents of leaves of S. luteo-caerulea in flowering stage of development. Materials and Methods: The essential oils were obtained by hydrodistillation of the leaves of S. luteo-caerulea and were analyzed by gas chromatography mass spectrometry (GC/MS). Moreover, microwave digestion with atomic absorption spectrophotometry were used for the mineral elements assay. Results: Ninety-seven constituents were detected. Between them, the major components were trans-caryophyllene (25.4%), D-germacrene (7.9%), and linalool (7.4%). Determination of mineral elements showed that the highest minerals were Ca2+ (65.14±1.95 µg/ml) and K+ (64.67±3.10 µg/ml). Conclusion: Presence of different essential oils and rich sources of Ca2+ and K+ candidate this plant as an auxiliary medication in different diseases, but more complementary researches are needed about its potency and side effects.

Background: Superparamagnetic iron oxide nanoparticles (SPION) have been largely considered for numerous applications in biomedicine such as magnetic resonance imaging, hyperthermia, cell tracking, anticancer treatment, and targeted delivery of drugs or genes. However, they may have side effects such body weight loss. Quercetin (QT), a strong antioxidant and free radical scavenger and a natural flavonoid, has a wide range of biological and therapeutic effects. In this study, the effect of QT on prevention of weight loss due to the using of SPION has been investigated. Materials and Methods: SPION and QT-SPION were administered orally at 50 and 100 mg/kg for 7 days. Then, the body weight was measured at the beginning and the end of the study. Results: Rats fed with 50 and 100 mg/kg SPION showed a significant weight loss, whereas those that fed with 50 mg/kg QT-SPION did not. A weight loss was observed in rats treated with 100 mg/kg of QT-SPION. Conclusions: The results of this study showed that quercetin could prevent weight loss due to the SPION.