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1.
Article in English | MEDLINE | ID: mdl-38984569

ABSTRACT

BACKGROUND: Iran has a relatively high prevalence of H. pylori, which correlates with high-risk areas for gastric cancer worldwide. METHODS: Our study aimed to investigate the underlying genetic mechanisms associated with resistance to metronidazole (frxA, rdxA), clarithromycin (23S rRNA), tetracycline (16S rRNA), and fluoroquinolone (gyrA) in H. pylori-positive dyspeptic patients using PCR and sequencing. We further examined the potential correlation between resistance profiles and various virulence genotypes. RESULTS: The rates of genetic mutations associated with resistance to metronidazole, fluoroquinolone, clarithromycin, and tetracycline were found to be 68%, 32.1%, 28.4%, and 11.1%, respectively. Well-documented multiple antibiotic resistance mutations were detected, such as rdxA and frxA (with missense and frameshift alterations), gyrA (Asp91, Asn87), 23S rRNA (A2142G, A2143G), and 16S rRNA (triple-base-pair substitutions AGA926-928→TTC). The cagA+ and vacA s1/m1 types were the predominant genotypes in our study. With the exception of metronidazole and tetracycline, no significant correlation was observed between the cagA+ and cagL+ genotypes and resistance-associated mutations. CONCLUSION: The prevalence of antibiotic resistance-associated mutations in H. pylori was remarkably high in this region, particularly to metronidazole, ciprofloxacin, and clarithromycin. By conducting a simultaneous screening of virulence and resistance genotypes, clinicians can make informed decisions regarding the appropriate therapeutic regimen to prevent the escalation of antibiotic resistance against H. pylori infection in this specific geographical location.

2.
Gastroenterol Hepatol Bed Bench ; 17(2): 151-160, 2024.
Article in English | MEDLINE | ID: mdl-38994502

ABSTRACT

Aim: The present study examined the protective potential of human adipose tissue-derived mesenchymal stem cells (hASCs) modified to overexpress alpha-1 antitrypsin (AAT), in a mouse model of the liver fibrosis. Background: For the treatment of end-stage liver diseases, cell therapy has emerged as a promising noninvasive alternative to liver transplantation. Mesenchymal stem cells (MSCs) are being evaluated due to their dual capabilities of promoting liver regeneration and modulating the pathogenic inflammation of the immune system. Methods: Liver fibrosis was induced in mice via the intraperitoneal injection of carbon tetrachloride (CCl4). MSCs were extracted from the human adipose tissue. After stemness confirmation, the cells were transduced with the lentiviruses containing the AAT gene, and then injected into the mice's tail vein. Fourteen days' post-transplantation, mice were sacrificed, and blood and tissue samples were collected for analysis. Important liver enzymes, including alanine transaminase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), albumin, and total bilirubin (TB), were measured. Histological studies were carried out using the hematoxylin and eosin (H&E), as well as Masson's trichrome (MT) staining. Results: Compared to hASCs, treatment with AAT-hASCs resulted in greater reductions in ALT, AST, ALP, and TB, as well as normalized albumin levels. AAT-hASCs promoted enhanced liver regeneration histologically, likely attributable to anti-inflammatory and anti-proteolytic properties of AAT. Conclusion: These findings indicate AAT-engineered hASCs as a promising cell-gene therapy candidate for further study in liver cirrhosis models.

3.
Endocrinol Diabetes Metab ; 7(1): e466, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38140923

ABSTRACT

INTRODUCTION: ANGPTLs (Angiopoietin-like proteins) 3 and 4 play an important role in the development of type 2 diabetes. These glycoproteins affect the modulation of glucose and lipid metabolism. They inhibit lipoprotein lipase (LPL) activity and provoke lipolysis. This study was aimed to investigate the protein levels of ANGPTL3 and 4 in the serum of type 2 diabetic patients with metabolic syndrome in comparison to the type 2 diabetic patients without metabolic syndrome and the control group. METHODS: Three groups of individuals were included in this study; Group I: 47 patients with type 2 diabetes and metabolic syndrome; Group II: 25 patients with type 2 diabetes without metabolic syndrome; Group III: 40 non-diabetic healthy people without metabolic syndrome as a control group. After collection of 5 mL fasting blood samples, serum concentrations of fasting blood sugar (FBS), cholesterol (Chol), triglyceride (TG), HDL-C (High-density lipoprotein-Cholesterol) and LDL-C (Low-density lipoprotein-Cholesterol) were measured by the enzymatic method; blood pressure (BP), height and weight with stadiometers; and ANGPTL3 and 4 by the enzyme-linked immunosorbent assay (ELISA). RESULTS: The serum levels of ANGPTL3 was significantly different among our three groups (p = .000). In patients with type 2 diabetes and metabolic syndrome (Group I), ANGPTL3 and 4 levels were lower than the control group. The serum levels of the parameters evaluated in this study (except HDL-C) was lower in the group II in comparison with the group I, and this difference was significant for TG, Chol, BP and BMI between these two groups. Also, our results revealed that there was a negative correlation between FBS, TG, Chol, LDL-C and BMI with ANGPTL3 and 4. While, there was a significant positive correlation between ANGPTL4 and ANGPTL3. CONCLUSION: Altogether, our findings suggest that the decreased levels of ANGPTL3 and 4 may be a causative factor for type 2 diabetes.


Subject(s)
Diabetes Mellitus, Type 2 , Metabolic Syndrome , Humans , Angiopoietin-Like Protein 3 , Diabetes Mellitus, Type 2/complications , Cholesterol, LDL , Control Groups , Triglycerides
4.
AMB Express ; 13(1): 112, 2023 Oct 13.
Article in English | MEDLINE | ID: mdl-37833506

ABSTRACT

Facilitated purification of proteins, at a low cost and a short time, is one of the key steps in the industrial production of recombinant proteins. In the current study, polydopamine nanoparticles (PDA-NPs) are considered in the synthesis of magnetic beads for purifying recombinant proteins due to advantages such as biocompatibility/ biodegradability, easy synthesis, as well as the ability to directly chelate metal ions. They were synthesized in Tris buffer (pH: 8:5), then chelated with Fe3+(20 mg) and Ni2+ ions at concentrations of 2, 3, 5, and 7 mg/ml. Prepared nanoparticles were characterized through scanning electron microscopy (SEM), ultraviolet-visible spectroscopy (UV-vis), dynamic light scattering (DLS), Inductively Coupled Plasma (ICP), and vibrating sample magnetometer (VSM). The size distribution of the particles was reported in the narrow range of 120-140 nm and 200 to 220 nm by the SEM image and DLS analysis, respectively. The chelation of ions on the surface of the nanoparticle was confirmed by the ICP technique with a magnetization of 35.42 emu/g. The highest adsorption rate of Ni2+ ions to polydopamine was obtained at a ratio of 1.4. The SDS-PAGE and western blot analysis confirmed the purification of eGFP and Hsp40 by PDA/Fe3+/Ni2+ at 26 and 40 kDa compared to the commercial nickel column. Moreover, the concentration of purified eGFP by PDA/Fe3+/Ni2+ was reported 138.83 µg/ml by the fluorescent signals, which is almost equal to or more than the protein purified by commercial Ni-NTA column (108.28 µg/ ml). The stability of PDA/Fe3+/Ni2+ has also been evaluated by ICP-OES for 10 days, and the result suggested that PDA magnetic beads were stable. Therefore, it can be concluded that PDA/Fe3+/Ni2+ have the ability to purify recombinant proteins in one less step and shorter time.

5.
Appl Microbiol Biotechnol ; 107(21): 6487-6496, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37672071

ABSTRACT

Today, enzymatic treatment is a progressive field in combating biofilm producing pathogens. In this regard, serratiopeptidase, a medicinally important metalloprotease, has been recently highlighted as an enzyme with proved anti-biofilm activity. In the present study, in order to increase the long-lasting effects of the enzyme, serratiopeptidase and the novel engineered forms with enhanced anti-biofilm activity were immobilized on the surface of cellulose nanofibers (CNFs) as a natural polymer with eminent properties. For this, recombinant serratiopeptidases including the native and previously designed enzymes were produced, purified and conjugated to the CNF by chemical and physical methods. Immobilization was confirmed using different scanning and microscopic methods. The enzyme activity was assessed using casein hydrolysis test. Enzyme release analysis was performed using dialysis tube method. Anti-biofilm activity of free and immobilized enzymes has been examined on Staphylococcus aureus and Pseudomonas aeruginosa strains. Finally, cytotoxicity of enzyme-conjugated CNFs was performed by MTT assay. The casein hydrolysis results confirmed fixation of all recombinant enzymes on CNFs by chemical method; however, inadequate fixation of these enzymes was found using cold atmospheric plasma (CAP). The AFM, FTIR, and SEM analysis confirmed appropriate conjugation of enzymes on the surface of CNFs. Immobilization of enzymes on CNFs improved the anti-biofilm activity of serratiopeptidase enzymes. Interestingly, the novel engineered serratiopeptidase (T344 [8-339ss]) exhibited the highest anti-biofilm activity in both conjugated and non-conjugated forms. In conclusion, incorporation of serratiopeptidases into CNFs improves their anti-biofilm activities without baring any cytotoxicity. KEY POINTS: • Enzymes were successfully immobilized on cellulose nanofibers using chemical method. • Immobilization of enzymes on CNFs improved their anti-biofilm activity. • T344 [8-339ss] exhibited the highest anti-biofilm activity in both conjugated and non-conjugated forms.


Subject(s)
Cellulose , Nanofibers , Cellulose/chemistry , Nanofibers/chemistry , Caseins , Biofilms
6.
Br J Haematol ; 201(4): 766-773, 2023 05.
Article in English | MEDLINE | ID: mdl-36477772

ABSTRACT

Many medications have been reported to be associated with thrombotic thrombocytopenic purpura (TTP) through pharmacovigilance data and published case reports. Whilst there are existing data available regarding drug-induced thrombotic microangiopathy, there is no available synthesis of evidence to assess drug-induced TTP (DI-TTP). Despite this lack of evidence, patients with TTP are often advised against using many medications due to the theoretical risk of DI-TTP. This systematic review evaluated the evidence for an association of medications reported as potential triggers for TTP. Of 5098 records available 261 articles were assessed further for eligibility. Fifty-seven reports, totalling 90 patients, were included in the final analysis. There were no cases where the level of association was rated as definite or probable, demonstrating a lack of evidence of any drug causing DI-TTP. This paucity of evidence was also demonstrated in the pharmacovigilance data, where 613 drugs were reported as potential causes of TTP without assessment of the strength of association. This systematic review demonstrates the need for standardised reporting of potential drugs causing TTP. Many reports omit basic information and, therefore, hinder the chance of finding a causative link if one exists.


Subject(s)
Purpura, Thrombotic Thrombocytopenic , Thrombotic Microangiopathies , Humans , Purpura, Thrombotic Thrombocytopenic/chemically induced , Pharmacovigilance , North America
7.
Iran Biomed J ; 26(6): 454-62, 2022 11 01.
Article in English | MEDLINE | ID: mdl-36437793

ABSTRACT

Background: Background: Hyaluronic acid (HA), a natural polymer with wide applications in biomedicine and cosmetics, is mainly produced by Streptococcal fermentation at industrial scale. In the present study, chemical random mutagenesis was used for development of Streptococcus equisimilis group G mutant strains with high HA productivity. Methods: Methods: The optimum of the pH of culture condition and cultivation time for HA production by wild strain group G were assessed. At first, two rounds of mutation at different concentrations of NTG was used for mutagenesis. Then, the nonhemolytic and hyaluronidase-negative mutants were screened on the blood and HA agar. HA productivity and molecular weight were determined by carbazole assay, agarose gel electrophoresis and specific staining. Moreover, stability of the high producer mutants was evaluated within 10 generations. Results: Results: The results showed that the wild-type strain produced 1241 ± 2.1 µg/ml of HA at pH 5.5 and 4 hours of cultivation, while the screened mutants showed a 16.1-45.5% increase in HA production. Two mutant strains, named Gm2-120-21-3 (2470 ± 8.1 µg/ml) and Gm2-120-21-4 (2856 ± 4.2 µg/ml), indicated the highest titer and a consistent production. The molecular weight (Mw) of HA for the mutants was less than 160 kDa, considering as a low Mw HA. Conclusion: Conclusion: The mutant strains producing a low polydisperse, as well as low Mw of HA with high titer might be regarded as potential industrial strains for HA production after further safety investigations.


Subject(s)
Hyaluronic Acid , Streptococcus , Hyaluronic Acid/chemistry , Molecular Weight , Agar
8.
AMB Express ; 12(1): 88, 2022 Jul 12.
Article in English | MEDLINE | ID: mdl-35821141

ABSTRACT

Hyaluronic Acid (HA) is a natural biopolymer that has important physiological and industrial applications due to its viscoelastic and hydrophilic characteristics. The responsible enzyme for HA production is Hyaluronan synthase (HAS). Although in vitro structure-function of intact HAS enzyme has been partly identified, there is no data on in vivo function of truncated HAS forms. In the current study, novel recombinant Bacillus subtilis strains harboring full length (RBSFA) and truncated forms of SeHAS (RBSTr4 and RBSTr3) were developed and HA production was studied in terms of titer, production rate and molecular weight (Mw). The maximum HA titer for RBSFA, RBSTr4 and RBSTr3 was 602 ± 16.6, 503 ± 19.4 and 728 ± 22.9 mg/L, respectively. Also, the HA production rate was 20.02, 15.90 and 24.42 mg/L.h-1, respectively. The findings revealed that RBSTr3 produced 121% and 137% more HA rather than RBSFA and RBSTr4, respectively. More interestingly, the HA Mw was about 60 kDa for all strains which is much smaller than those obtained in prior studies.

9.
Chem Biol Drug Des ; 100(4): 553-563, 2022 10.
Article in English | MEDLINE | ID: mdl-35729860

ABSTRACT

The clinical application of serratiopeptidase as an anti-biofilm and anti-inflammatory agent is restricted due to the enzyme sensitivity to the environmental conditions. In our previous study, six enzyme variants were designed by introducing different mutations and truncations that exhibited higher thermal stability. In the present study, the interaction pattern and affinity of variants to substrates and inhibitors were studied using molecular docking and in vitro studies. CABS-dock and Swiss-dock servers were used for substrate (Bradykinin and Substance-P) and inhibitor (Lisinopril and EDTA) docking, respectively. The interactions were analyzed using LigPlot, UCSF Chimera, and visual molecular dynamics packages. Free energy calculations were performed using PRODIGY. Finally, the native enzyme and the best variant in terms of interaction pattern and binding score were selected for in-vitro affinity analysis toward Bradykinin and EDTA using HPLC and casein hydrolysis test, respectively. Molecular docking revealed that T344 [8-339ss] variant showed a different pattern for both substrates and inhibitors in the way that none of the native active site residues were involved in the receptor binding. As revealed by in vitro studies, T344 [8-339ss] displayed the highest number of hydrogen bond formation in docking with Bradykinin and remarkable decrement in the binding affinity for EDTA. This was the first report on the design of novel serratiopeptidase with higher activity to Bradykinin and improved resistance to EDTA as an inhibitor.


Subject(s)
Bradykinin , Caseins , Anti-Inflammatory Agents , Edetic Acid , Hydrogen Bonding , Hydrophobic and Hydrophilic Interactions , Lisinopril , Molecular Docking Simulation , Molecular Dynamics Simulation , Peptide Hydrolases
10.
Biol Trace Elem Res ; 200(4): 1562-1567, 2022 Apr.
Article in English | MEDLINE | ID: mdl-34195940

ABSTRACT

BACKGROUND: Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is the cause of the COVID-19 pandemic and is the cause of increased mortality, especially among elderly patients and those who have severe complications, such as chronic pulmonary obstruction, hypertension, diabetes, and cancer. Nutrition, especially micronutrients, plays an important role in reducing mortality and complications from COVID-19 because micronutrients strengthen our immune system and nutritional status is an important factor that affects the outcome of patients with COVID-19. Among micronutrients, selenium has an important effect on both intrinsic and acquired immunity. Host selenium deficiency affects the viral genome and increases the virulence of viruses. We have investigated the serum selenium levels in COVID-19 patients and healthy control individuals. METHODS: A total of 50 patients with COVID-19 infection were included in this study. During hospitalization, 13 patients died (non-survivor group) and 37 patients recovered (survivor group). We assessed the serum selenium levels in 50 COVID-19 patients and 50 healthy individuals by Agilent SpectrAA-240 Z atomic absorption spectrometer. RESULTS: The serum selenium level was significantly lower in COVID-19 patients (77. 8 ± 13.9 µg/L) as compared to healthy control individuals (91.7 ± 16.7 µg/L), but there was no significant difference between the survivor and non-survivor groups. Also, there was no significant relationship between serum selenium levels and laboratory findings of COVID-19 patients. CONCLUSIONS: These results suggest that decreased serum selenium levels may be a risk factor for the COVID-19 infection, but there was no significant relationship between selenium and severity and mortality of COVID-19 disease.


Subject(s)
COVID-19 , Selenium , Aged , Humans , Micronutrients , Pandemics , SARS-CoV-2
11.
Middle East J Dig Dis ; 14(4): 396-403, 2022 Oct.
Article in English | MEDLINE | ID: mdl-37547507

ABSTRACT

Background: Esophageal cancer is one of the main causes of cancer mortality in the world. Golestan province, in the northern part of Iran, has the highest esophageal cancer rate in the world. The north and south districts of Golestan province can be classified as low and high-risk areas for esophageal cancer. One of the potential risk factors for esophageal cancer in this population is a nutrient-deficient diet. Dietary antioxidant compounds such as selenium, vitamin E, vitamin A, and ß-carotene are reactive oxygen species (ROC) scavengers that play a key role in cellular responses to oxidative stress and preventing DNA damage. This study aims to compare the serum levels of selenium, vitamin E, and vitamin A in healthy individuals in high and low-risk areas of esophageal cancer. Methods: This study is a population of 242 healthy individuals. Serum selenium levels were assessed by atomic absorption spectroscopy. Vitamin E and A were assessed by reversed-phase high-performance liquid chromatography. Results: Vitamin E levels of healthy individuals in high-risk areas were significantly lower than in low-risk areas, while there was no significant difference between the selenium and vitamin A levels of healthy individuals in high-risk areas and low-risk areas. Also, there was no significant difference between selenium, vitamin E, and vitamin A levels in urban and rural areas and men and women in Golestan province. Conclusion: High levels of selenium with lower levels of vitamin E, along with other risk factors, may be associated with esophageal squamous cell carcinoma in high-risk areas of Golestan province.

12.
Biomater Sci ; 8(13): 3611-3627, 2020 Jul 07.
Article in English | MEDLINE | ID: mdl-32515439

ABSTRACT

The limited regenerative capacity of the CNS poses formidable challenges to the repair of spinal cord injury (SCI). Two key barriers to repair are (i) the physical gap left by the injury, and (ii) the inhibitory milieu surrounding the injury, the glial scar. Biomaterial implantation into the injury site can fill the cavity, provide a substrate for cell migration, and potentially attenuate the glial scar. We investigated the biological viability of a biocompatible and biodegradable poly-ε-lysine based biomaterial, Proliferate®, in low and high cross-linked forms and when coated with IKVAV peptide, for SCI implantation. We demonstrate altered astrocyte morphology and nestin expression on Proliferate® compared to conventional glass cell coverslips suggesting a less reactive phenotype. Moreover Proliferate® supported myelination in vitro, with myelination observed sooner on IKVAV-coated constructs compared with uncoated Proliferate®, and delayed overall compared with maintenance on glass coverslips. For in vivo implantation, parallel-aligned channels were fabricated into Proliferate® to provide cell guidance cues. Extensive vascularisation and cellular infiltration were observed in constructs implanted in vivo, along with an astrocyte border and microglial response. Axonal ingrowth was observed at the construct border and inside implants in intact channels. We conclude that Proliferate® is a promising biomaterial for implantation following SCI.


Subject(s)
Biocompatible Materials/chemistry , Central Nervous System Diseases/therapy , Polylysine/chemistry , Prostheses and Implants , Spinal Cord Injuries/therapy , Animals , Biocompatible Materials/chemical synthesis , Cells, Cultured , Polylysine/chemical synthesis , Rats , Rats, Sprague-Dawley
13.
Asian Pac J Cancer Prev ; 20(10): 2897-2901, 2019 Oct 01.
Article in English | MEDLINE | ID: mdl-31653132

ABSTRACT

INTRODUCTION: Esophageal cancer is one of the most lethal gastrointestinal cancers that has a complex and diverse etiology, with several genetic and nutritional factors involved in its etiology. The purpose of this study was to investigate the type of haptoglobin genotype and its relationship with some nutritional and biochemical risk factors affecting the prevalence of esophageal cancer in patients with early stage esophageal cancer. MATERIALS AND METHODS: In this study, 44 patients (20 males and 24 females) with early stage esophageal cancer and 44 healthy subjects, classified as control group, (19 males and 25 females) were selected. Haptoglobin (HP) genotype was determined employing PCR technique. Nutritional data were analyzed using standard food frequency questionnaire (FFQ) method. Serum levels of malondialdehyde (MDA), nitrate and nitrite were measured employing the colorimetric method. Serum levels of p53 protein were measured using the enzyme-linked immunosorbent assay (ELISA) technique. RESULTS: The results of our study showed for the first time that HP1-1 genotype was the most prevalent genotype in esophageal cancer patients in Golestan province, Iran. HP2-2 genotype was the most frequent in the control group. Serum levels of MDA were significantly higher in the patients' group compared to the control group (P˂0.001). Weight and body mass index (BMI) were significantly lower in the patients' group than the control group (P<0.01). Food frequency analysis revealed that the consumption of fruits and vegetables in the patients' group was lower than that of the control group (P<0.05). CONCLUSION: The results of our study showed for the first time that HP1-1 genotype is the dominant genotype in patients with esophageal cancer in Golestan province. As well, modification of nutritional pattern and consumption of high level of antioxidant compounds can be effective in reducing the prevalence of esophageal cancer in this region.


Subject(s)
Esophageal Neoplasms/epidemiology , Esophageal Neoplasms/genetics , Haptoglobins/genetics , Nutritional Status , Polymorphism, Genetic , Aged , Case-Control Studies , Esophageal Neoplasms/pathology , Female , Follow-Up Studies , Genotype , Humans , Iran/epidemiology , Male , Middle Aged , Neoplasm Staging , Prevalence , Risk Factors
14.
Exp Neurol ; 283(Pt B): 541-9, 2016 09.
Article in English | MEDLINE | ID: mdl-26988764

ABSTRACT

Astrocytes are the major glial cell of the central nervous system (CNS), providing both metabolic and physical support to other neural cells. After injury, astrocytes become reactive and express a continuum of phenotypes which may be supportive or inhibitory to CNS repair. This review will focus on the ability of astrocytes to influence myelination in the context of specific secreted factors, cytokines and other neural cell targets within the CNS. In particular, we focus on how astrocytes provide energy and cholesterol to neurons, influence synaptogenesis, affect oligodendrocyte biology and instigate cross-talk between the many cellular components of the CNS.


Subject(s)
Astrocytes/physiology , Myelin Sheath/physiology , Animals , Astrocytes/chemistry , Cell Differentiation , Central Nervous System , Cytokines/metabolism , Humans , Neurons/physiology
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