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The use of anticancer peptides (ACPs) as an alternative/complementary strategy to conventional chemotherapy treatments has been shown to decrease drug resistance and/or severe side effects. However, the efficacy of the positively-charged ACP is inhibited by elevated levels of negatively-charged cell-surface components which trap the peptides and prevent their contact with the cell membrane. Consequently, this decreases ACP-mediated membrane pore formation and cell lysis. Negatively-charged heparan sulphate (HS) and chondroitin sulphate (CS) have been shown to inhibit the cytotoxic effect of ACPs. In this study, we propose a strategy to promote the broad utilization of ACPs. In this context, we developed a drug repositioning pipeline to analyse transcriptomics data generated for four different cancer cell lines (A549, HEPG2, HT29, and MCF7) treated with hundreds of drugs in the LINCS L1000 project. Based on previous studies identifying genes modulating levels of the glycosaminoglycans (GAGs) HS and CS at the cell surface, our analysis aimed at identifying drugs inhibiting genes correlated with high HS and CS levels. As a result, we identified six chemicals as likely repositionable drugs with the potential to enhance the performance of ACPs. The codes in R and Python programming languages are publicly available in https://github.com/ElyasMo/ACPs_HS_HSPGs_CS. As a conclusion, these six drugs are highlighted as excellent targets for synergistic studies with ACPs aimed at lowering the costs associated with ACP-treatment.

The use of anticancer peptides (ACPs) as an alternative/complementary strategy to conventional chemotherapy treatments has been shown to decrease drug resistance and/or severe side effects. However, the efficacy of the positively-charged ACP is inhibited by elevated levels of negatively-charged cell-surface components which trap the peptides and prevent their contact with the cell membrane. Consequently, this decreases ACP-mediated membrane pore formation and cell lysis. Negatively-charged heparan sulphate (HS) and chondroitin sulphate (CS) have been shown to inhibit the cytotoxic effect of ACPs. In this study, we propose a strategy to promote the broad utilization of ACPs. In this context, we developed a drug repositioning pipeline to analyse transcriptomics data generated for four different cancer cell lines (A549, HEPG2, HT29, and MCF7) treated with hundreds of drugs in the LINCS L1000 project. Based on previous studies identifying genes modulating levels of the glycosaminoglycans (GAGs) HS and CS at the cell surface, our analysis aimed at identifying drugs inhibiting genes correlated with high HS and CS levels. As a result, we identified six chemicals as likely repositionable drugs with the potential to enhance the performance of ACPs. The codes in R and Python programming languages are publicly available in https://github.com/ElyasMo/ACPs_HS_HSPGs_CS. As a conclusion, these six drugs are highlighted as excellent targets for synergistic studies with ACPs aimed at lowering the costs associated with ACP-treatment., QC 20220608

The production of polysaccharides derived from lactic acid bacteria (LAB) can be a valuable alternative to current polysaccharides. In this study, eight samples of Kishk (traditional dairy product) were collected in sterile conditions and directly cultured on MRS agar medium. Following purification and examination of microscopic, macroscopic characteristics and doing biochemical tests, 143 isolates were selected, and the production of exopolysaccharide (EPS) was investigated by the ruthenium red and disc methods. The EPS production of 79 isolates was confirmed. Total carbohydrate of EPS was determined by the phenol sulfuric acid method. Finally, Enterococcus durans K48, Enterococcus faecium R114, and Enterococcus faecium T52 strains were selected as the best EPS producers. The optimization of EPS production was then performed using Central Composite Design (CCD) and Response Surface Methodology (RSM) approaches. Optimization plots showed the highest EPS yield for E. durans K48, E. faecium R114, and E. faecium T52 occurred at 38.4, 37.4 and 36.7 °C and pH of 5.9, 5.6 and 5.8, respectively, cultured in a Sucrose-MRS medium. Under optimal conditions, the maximum predicted and actual production of EPSs for the examined isolates were 3.18, 3.21, 2.99 and 3.02, 3.15, 3.15, g L, respectively. As determined by Gel Permeation Chromatography, the EPSs molecular weights were in the range of 2.93 × 10 to 3.52 × 10 Da. Analysis of the component monosaccharides by HPLC showed that all three tested EPSs were heteropolysaccharides. After appropriate evaluation, the EPSs produced by native species of LAB isolated from Iranian Kishk could be of interest for industrial applications or as functional food ingredients.

Exopolysaccharides are high molecular weight polymers composed of sugar subunits. Produced exopolysaccharides by lactic acid bacteria (LAB) play a significant role in improvement of organoleptic properties of fermented dairy products such as yogurt. Diversely, the probiotic function of these bacteria and the prebiotic properties of their produced biopolymers promote consumer¿s health. For this purpose, a traditional dairy product known as ¿Kishk¿ was selected. 143 strains of lactic acid bacteria were isolated from Iranian Kishk in Khorasan Province and cultured in formulated MRS mediums with different sugars such as glucose, fructose, sucrose and, lactose (40 g/L) and incubated in anaerobic conditions at 30 and 37°C for 48 hours. The microscopic features of the isolates were assessed and the production of exopolysaccharide in the culture medium was evaluated by disk and ruthenium red methods. The phenol-sulfuric and weight method were used to quantify exopolysaccharide production. Results showed pH of Kishk samples ranged from 3.60 to 4.08 and the average of total mesophilic count and LAB count of samples were 6.50 and 5.89 log CFU/g, respectively. Analysis of data exhibited 79 out of 143 lactic acid bacteria isolates were exopolysaccharide producer and 70% of them were cocci. The average of maximum and minimum production by weight method were 2.61 g/L and 0.08 g/L, respectively. The average of highest and the lowest amount of exopolysaccharide by phenol sulfuric method were measured 1.87 g/L and 0.06 g/L, respectively. This study indicates the potential of exopolysaccharide production by Iranian native species from dairy products.

In this study, the antimicrobial effect of exopolysaccharide (EPS) extracted from Enterococcus strains [E. durans K48 (MT437,248), E. faecium R114 (MT437,249) and E. faecium T52 (MT437,250)] isolated from Kishk was applied against some foodborne pathogenic bacteria using well diffusion and microdilution methods. The antioxidant activity of EPS was also evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay and ferric reducing antioxidant power (FRAP) method. The cytotoxicity effect of EPS on human Gingival Fibroblast (HGF) cell line was also assessed. The results obtained by antimicrobial test showed that the most resistant bacteria to the examined EPS was Listeria monocytogenes, and the most susceptible were Staphylococcus aureus and E. faecalis. The results showed that the DPPH inhibitory percentage of EPS (25 mg/mL) from E. durans K48, E. faecium R114, and E. faecium T52 was 53%, 58% and 64%, respectively. EPS from E. faecium T52 displayed the highest reducing power, but statistically, there was no significant difference between the reducing power of EPS T52 and EPS R114 (P ≥ 0.05). The lowest toxicity percentage of EPS k48, EPS T52, and EPS R114 on normal human cell line at a concentration of 0.2 mg/mL was 10%, 15%, and 13%, respectively, which was statistically significant (P < 0.05). The obtained results in the present study indicate that EPS from the examined LAB strains with no in vitro cytotoxicity can be a potential source of natural antioxidant and antibacterial agent to be used in food and pharmaceutical industries.

High levels of growth hormone accelerate mitosis rate but decrease the apoptosis process in its target organs. These events might cause the initiation of different cancer types. Thus, the main aims of this study were assessing the effects of short term growth hormone administration and resistance training on the histopathology and detection of the BRAF-V600E mutation in the thyroid tissue of male Rattus norvegicus brown rats. Thirty-two rats were randomly divided into four groups. After 8 weeks of the experiment (i.m), thyroid tissue and blood samples of saline (CS), resistance training+saline (RS), growth hormone (2 mg/kg) (GI) and resistance training+growth hormone (2 mg/kg) (RG) were taken to evaluate histopathology, the BRAF T1799A mutation of thyroid tissue, and circulating levels of IGF-1 and IGFBP-3. The protocol of training consisted of rats climbing a ladder while carrying weights (3 sets/5 reps). Microscopic evaluation of thyroid tissue did not show any histopathological changes, and there were no mutations in the studied region of the BRAF sequence. Serum IGF-1 concentration was significantly lower in the RS group than in other groups (P<0.05). However, serum IGFBP-3 concentration did not change significantly in the RS group. Moreover, serum IGF-1 and IGFBP-3 concentrations were significantly higher in the GI and RG groups than in the others (P<0.05). In conclusion, the decrement of serum IGF-1 concentration and IGF-1/IGFBP-3 ratio after resistance training might decrease the risk of cancer. Furthermore, short term growth hormone administration, with and without resistance training, might increase the risk of cancer through the high levels of serum IGF-1 concentration and IGF-1/ IGFBP-3 ratio in male rats., Visoke razine hormona rasta povećavaju brzinu mitoze, ali smanjuju proces apoptoze u ciljnim organima, što može uzrokovati nastanak različitih tipova raka. Stoga je glavni cilj ovoga istraživanja bio utvrditi učinak kratkotrajne primjene hormona rasta i treninga snage na histopatološke promjene tkiva štitnjače i nalaz mutacije gena BRAF-V600E u mužjaka smeđeg štakora Rattus norvegicus. Ukupno 32 štakora nasumično su podijeljena u četiri skupine koje su ovisno o primjenjenom tretmanu označene kao CS (fiziološka otopina), RS (trening snage + fiziološka otopina), GI (hormone rasta, 2 mg/kg) i RG (trening snage+hormone rasta, 2 mg/kg). Nakon osam tjedana pokusa (i.m.) uzeti su uzorci tkiva štitnjače i uzorci krvi kako bi se procijenila histopatološka svojstva, mutacija BRAF T1799A tkiva štitnjače te razine cirkulacijskog IGF-1 i IGFBP-3. Trening se sastojao od toga da se štakori penju ljestvama noseći težinu (3 seta vježbi, 5 ponavljanja). Mikroskopska procjena tkiva štitnjače nije pokazala histopatološke promjene i nije bilo mutacija u promatranoj regiji sekvencija gena BRAF. Koncentracija serumskog IGF-1 bila je znakovito manja u skupini RS nego u ostalim skupinama (P < 0,05), no u toj skupini nije bilo znakovite promjene u koncentraciji serumskog IGFBP-3. Štoviše, koncentracije serumskog IGF-1 i IGFBP-3 bile su znakovito veće u skupinama GI i RG nego u ostalim skupinama (P < 0,05). Stoga smo zaključili da bi smanjenje koncentracije serumskog IGF-1 i IGF-1/IGFBP-3 nakon treninga snage moglo smanjiti rizik od raka. Osim toga kratkotrajna primjena hormona rasta, s treningom snage kao i bez njega, mogla bi povećati rizik od raka povećavajući koncentracije serumskog IGF-1 i IGF-1/IGFBP-3 u mužjaka štakora.

The insulin-like growth factor (IGF) axis is fundamental for mammalian growth and development. However, no comprehensive reference data on gene expression across tissues and pre- and postnatal developmental stages are available for any given species. Here we provide systematic promoter- and splice variant specific information on expression of IGF system components in embryonic (Day 48), fetal (Day 153), term (Day 277, placenta) and juvenile (Day 365–396) tissues of domestic cow, a major agricultural species and biomedical model. Analysis of spatiotemporal changes in expression of IGF1, IGF2, IGF1R, IGF2R, IGFBP1-8 and IR genes, as well as lncRNAs H19 and AIRN, by qPCR, indicated an overall increase in expression from embryo to fetal stage, and decrease in expression from fetal to juvenile stage. The stronger decrease in expression of lncRNAs (average ―16-fold) and ligands (average ―12.1-fold) compared to receptors (average ―5.7-fold) and binding proteins (average ―4.3-fold) is consistent with known functions of IGF peptides and supports important roles of lncRNAs in prenatal development. Pronounced overall reduction in postnatal expression of IGF system components in lung (―12.9-fold) and kidney (―13.2-fold) are signatures of major changes in organ function while more similar hepatic expression levels (―2.2-fold) are evidence of the endocrine rather than autocrine/paracrine role of IGFs in postnatal growth regulation. Despite its rapid growth, placenta displayed a more stable expression pattern than other organs during prenatal development. Quantitative analyses of contributions of promoters P0-P4 to global IGF2 transcript in fetal tissues revealed that P4 accounted for the bulk of transcript in all tissues but skeletal muscle. Demonstration of IGF2 expression in fetal muscle and postnatal liver from a promoter orthologous to mouse and human promoter P0 provides further evidence for an evolutionary and developmental shift from placenta-specific P0-expression in rode

The insulin-like growth factor (IGF) axis is fundamental for mammalian growth and development. However, no comprehensive reference data on gene expression across tissues and pre- and postnatal developmental stages are available for any given species. Here we provide systematic promoter- and splice variant specific information on expression of IGF system components in embryonic (Day 48), fetal (Day 153), term (Day 277, placenta) and juvenile (Day 365–396) tissues of domestic cow, a major agricultural species and biomedical model. Analysis of spatiotemporal changes in expression of IGF1, IGF2, IGF1R, IGF2R, IGFBP1-8 and IR genes, as well as lncRNAs H19 and AIRN, by qPCR, indicated an overall increase in expression from embryo to fetal stage, and decrease in expression from fetal to juvenile stage. The stronger decrease in expression of lncRNAs (average ―16-fold) and ligands (average ―12.1-fold) compared to receptors (average ―5.7-fold) and binding proteins (average ―4.3-fold) is consistent with known functions of IGF peptides and supports important roles of lncRNAs in prenatal development. Pronounced overall reduction in postnatal expression of IGF system components in lung (―12.9-fold) and kidney (―13.2-fold) are signatures of major changes in organ function while more similar hepatic expression levels (―2.2-fold) are evidence of the endocrine rather than autocrine/paracrine role of IGFs in postnatal growth regulation. Despite its rapid growth, placenta displayed a more stable expression pattern than other organs during prenatal development. Quantitative analyses of contributions of promoters P0-P4 to global IGF2 transcript in fetal tissues revealed that P4 accounted for the bulk of transcript in all tissues but skeletal muscle. Demonstration of IGF2 expression in fetal muscle and postnatal liver from a promoter orthologous to mouse and human promoter P0 provides further evidence for an evolutionary and developmental shift from placenta-specific P0-expression in rode