Archives of Razi Institute最新文献

The current experimental study is designed to examine the in vitro and in vivo effects of green synthesized silver nanoparticles (AgNPs) against Giardia lamblia, a major cause of parasitic diarrhea. The precipitation method was employed for the green synthesis of AgNPs by Astragalus ecbatanus aqueous extract. In the, in vitro, Giardia lamblia cysts and trophozoites were exposed to AgNPs at 10, 20, and 40 mg/mL for 10-360 min. The effects of AgNPs on trophozoite plasma membrane and their cytotoxic effects on normal and colon cancer cells were evaluated using Sytox green and MTT assay for cell viability. The in vivo assay included BALB/c mice, infected by Giardia, treated with AgNPs at 10, 15, and 20 mg/kg/day for one week. On the 8th day post-infection, stool examination was conducted to assess the presence of Giardia cysts and the reduction rate. The size distribution of AgNPs ranged between 5 and 80 nm, with the maximum particle size observed at 40-60 nm. AgNPs significantly (P<0.001) increased the mortality of Giardia lamblia trophozoites in a dose-dependent manner. Specifically, AgNPs at concentrations of 200 and 300 μg/mL destroyed Giardia lamblia cysts after 4 and 2 h, respectively. Trophozoites of Giardia lamblia showed more sensitivity to AgNPs compared to cysts. At concentrations of 100, 200, and 300 μg/mL, AgNPs eliminated all trophozoites after 4, 2, and 1 h of treatment, respectively. AgNPs dose-dependently reduced (P<0.001) the parasite load and viability of Giardia lamblia cysts. Exposure of Giardia lamblia trophozoites to AgNPs dose-dependently increased the plasma membrane permeability as indicated by rise in the exposed fluorescence. The CC₅₀ value AgNPs for colon cancer and normal cell lines was 402.3 μg/mL and 819.6 μg/mL, respectively. The selectivity value greater than 2 (2.04), suggests that these AgNPs are safe for normal cells in comparison with cancer cells. This experimental study showed that AgNPs green synthesized by Astragalus ecbatanus exhibited significant in vitro and in vivo anti-Giardia activity, positioning them as potential candidates for Giardia infection treatment. Nevertheless, further research on the precise mechanisms of action and comprehensive exploration of all toxicity aspects associated with this type of AgNPs need to be considered.

One of the major roles of nanotechnology in the pharmaceutical field is to provide a facility to improve drug delivery systems and design smart nanocarriers with the potential to deliver specific biomolecules to the target site for treatment. This study evaluated Sonchus maritimus-loaded niosomes (SmE-N) in hepatic encephalopathy induced by a high-fructose diet (HFD) in rats. High-performance liquid chromatography (HPLC) analysis of Sonchus maritimus extracts (SmE), the synthesis of niosomes, and their characterization were performed. For the in vivo study, 24 male rats were haphazardly divided into 4 groups (n=6) control, HFD (35%), HFD+SmE-N (50 mg/kg/day), and HFD+metformin (50 mg/kg/day). Clinical behaviors and biological markers were assessed for all groups. The in vitro results of the chromatographic analysis revealed that Sonchus maritimus contains important phenolic acids, including gallic acid, vanillic acid, chlorogenic acid, and caffeic acid, as well as diverse flavonoids, including quercetin, rutin, and naringin bioactive compounds. The niosome formulation, characterized by the encapsulation efficiency of SmE, reached up to 61.40%. The in vivo results of the HFD showed a significant change in behavior parameters, liver glycogen, transaminase enzymes, brain protein, and acetylcholine esterase levels. In addition, there was a significant increase in malondialdehyde levels and a decrease in glutathione, superoxide dismutase, and glutathione peroxidase activities in the HFD group compared to the control group. Furthermore, the histopathological observation recorded a profound modification in the liver and brain tissues of the HFD group. In contrast, the treatment with SmE-N and metformin assured a partial amelioration in the noticed parameters compared to the HFD group, but SmE-N led to a better improvement than metformin compared to the control group. In conclusion, the use of SmE-N bioconjugated by linoleic acid seems powerful in treating the complications of fructose-induced metabolic disorders due to its hepato-neuroprotective abilities.

The BCG vaccines on the market have employed a Mycobacterium bovis (M. bovis) sub-strains derived from the initial strain. To date, there has been no recommendation regarding the sub-strains with the highest effectiveness when administered to humans. Because it remains the standard for Tuberculosis treatment, the quality of the BCG vaccine must be verified. The viability test is one of the parameters for BCG vaccine quality control. The culture method has become the gold standard for viability testing with various testing media. The present study aimed to evaluate the performance of Lowenstein Jensen (LJ) and Ogawa media for the viability test of Pasteur 1173P2 and Russian (Moscow) - 384 sub-strains of M. bovis in the BCG vaccine. The number of culturable particles of each sub-strain in the BCG vaccine was estimated and statistically evaluated using the t-test. The colonies of the Pasteur 1173P2 have characteristics; tended to clump on both mediums with tiny, rough, and pale yellow/cream colors. Although the colony character of the Russian (Moscow) - 384 generally has similar feature, it did not cluster and had a smooth texture. In terms of growth rate, LJ and Ogawa media performed similarly for Pasteur 1173P2 and Russian (Moscow) - 384 sub-strains. Maximum growth is reached by the fifth week. The culturable particles of Pasteur P1173P2 sub-strains did not differ between mediums. Whereas the growth of the Russian (Moscow) - 384 sub-strains was statistically better on Ogawa media. The results of this study reveal that the performance of the media used for determining the number of culturable particles is based on the sub-strains of M. bovis present in the BCG vaccine.

Campylobacter spp. genera is one of the most common causes of microbial enteritis worldwide. This study aimed to find out how common Campylobacter organisms were in raw meat from large livestock in Iran, as well as to determine their antibiotic susceptibility profiles. Several 550 fresh, ready-to-eat meat samples were collected from slaughterhouses, butcher shops, and restaurants in the study region. The samples were collected from cattle (n=138), goats (n=102), camels (n=56), and sheep (n=254). Campylobacter spp. were isolated and identified using normal bacteriological methods and polymerase chain reaction (PCR). Genotyping was performed using PCR to identify virulence genes. The disc diffusion technique was used to determine antibiotic susceptibility. The two Campylobacter spp. were found in 84 (15.27%) of the 550 meat samples tested. Cattle and camel samples accounted for the highest (52.38%) and lowest (3.57%) frequencies of Campylobacter spp., respectively. There were significant differences in the prevalence of Campylobacter spp. in cattle (2=43.04 or OR=7.68, CI=3.40-17.30, P<0.01). Campylobacter jejuni and Campylobacter coli accounted for 82.14% (n=69) of Campylobacter spp. isolated from raw meat. While C. jejuni was found in 39.28% of the samples (n=33), C. coli was observed in 42.85% (n=36). Other Campylobacter spp. formed 17.85 % (n=15) of the samples. The most common genotypes observed in C. jejuni bacteria collected from different types of large animal samples were ciaB (100%) and flaA (100%). On the other hand, virbll (7.69%) was the C. jejuni strain found with the lowest incidence in different large animal samples. The most frequent genotypes found in C. coli bacteria were ciaB (100%) and flaA (100%). C. coli isolates dnaJ (0%), wlaN (0%), virbll (0%), and ceuE (0%) were detected with the lowest frequency in several samples from large livestock. Campylobacter spp. isolated from different sample types and sources were 100% sensitive to aphA-3-1 and GM10. The isolates were reported to be resistant to E15 (76.93%), cmeB (69.24%), aadE1 (69.24%), CIP5 (69.24%), and AM10 (69.24%). According to this study, Campylobacter was found in food from factory farming. Consequently, the disease can be transmitted by eating raw or undercooked meat. Therefore, proper handling and preparation of meat meals, as well as hygiene measures from the slaughterhouse to the retailer, are critical in preventing Campylobacter infections.

Hepatitis B virus (HBV) X protein (HBx) plays a key role in hepatocellular carcinoma (HCC). HBx may alter the expression of multiple microRNAs (miRs), which are important in hepatocarcinogenesis. This study aimed to investigate the importance of HBx protein in the expression of miR-21, miR-22, miR-122, miR-132, and miR-222. A recombinant vector expressing HBx was developed. The Huh-7 cell line was transfected with the HBx-pcDNA3.1+ recombinant plasmid. A Real-Time Polymerase Chain Reaction was used to evaluate the expression of miR-21, miR-22, miR-122, miR-132, and miR-222 in the cell line. It was found that the expression of miR-21 and miR-222 was upregulated at all points of time after HBx transfection. The expression of miR-21 was 4.24-fold 72 h after transfection. The miR-22 had a 7.69-fold downregulation after 24 h, and the miR-122 had a significant downregulation after 48 h (10-fold). The miR-132 expression reached its lowest rate 12 h after HBx transfection (8.33-fold), and the miR-222 expression was upregulated in transfected cells but was not significantly different (1.18- to 2.45-fold). The significant downregulation of miR-22, miR-122, and miR-132 implicates their inhibitory roles in the progression of HBV-associated HCC. The expression of these microRNAs could be used as a prognostic marker for the progression of HBV-associated liver disease.

The appearance of an array of data on the study of the intestinal microbiota in Metazoa has significantly expanded our understanding of the role of commensals in the control of a wide range of physiological functions in higher organisms in norm and pathology. In the intestine, where the microbial load significantly exceeds the number of microorganisms of other ecosystems, the components of the intestinal microbiota are a constant source of stimuli that induce activation of the host immune system. The introduction into practice of biomedical research of innovative high-resolution methods, including   multi-omics technologies, has brought data that change our understanding of intestinal commensals, including probiotics with GRAS status, widely used in medicine, agriculture and biotechnology. The ability of these bacteria to induce negative processes in the host body that are beneficial for bacterial proliferation and expansion revealed a clear lack of our knowledge about the logic of their life and the mechanisms of interaction with eukaryotic cells. This determines the urgent need for comprehensive research of probiotics and the development of standardization of their safety assessment. Apriori's confidence in the exceptional benefit of the bacteria widely used in medicine, agriculture and biotechnology has determined the seriously omission in our control system today - the lack of standardization of studies for the safety assessment of bacteria with GRAS status . The moment has come when it became clear that this gap should be promptly filled and that only exact understanding the molecular base of interacting the microbes with eukaryotic cells can provide the foundation for effective practical developments in controlling the evolution of bacterial virulence and probiotic safety strategy, as well as the competent use of genetic technologies for monitoring the environment and managing infectious processes, thus avoiding the dramatic consequences of large-scale interventions in the micro and macro worlds.

Selenium is a class 2B element according to the International Council for Harmonization Q3D guidelines. Selenium sulfide is an anti-infective agent with antifungal and antibacterial properties used to treat dandruff and seborrheic dermatitis. The literature survey revealed that most of the analytical techniques to estimate selenium were time-consuming and/or required high skill levels. The process involved identifying the isotopes, selecting the measurement approach, and optimizing a typical microwave-aided digesting procedure. Ammonium hydrogen difluoride, water, and concentrated nitric acid were added to the samples. The confirmed microwave digestion program was a two-step program where in the initial step, the samples were ramped at 200°C for 20 min and held for 5 min. Later, samples were cooled and neutralized by boric acid, then ramped for 20 min to a temperature of 180°C and held for 10 min. Selenium was estimated at 196.090 nm by inductively coupled plasma optical emission spectroscopy (ICP-OES). System suitability was run before initiating analysis to ensure that system performance was consistent. Analytical validation parameters, such as the specificity of the method, were demonstrated at 196.090 nm, linearity was proven from 10 ppm to 150 ppm of selenium concentration, the detection limit was 1.28 ppm, and the limit of quantification was 3.89 ppm. Robustness was confirmed for small changes to ICP-OES operating conditions. The precision of the method demonstrated by analyzing the percentage relative standard deviation for six injections was found to be less than 2.0%. Accuracy was confirmed from 10 ppm to 150 ppm, and all the samples were observed to be within the range of 95%-105%. A common microwave-assisted digestion technique was developed and validated as well. The precision, specificity, linearity, accuracy, and robustness of the method for estimating selenium in selenium sulfide drug substances and various pharmaceutical dosage forms were demonstrated. This newly developed microwave-assisted digestion technique has optimum sensitivity and is highly reproducible and time-saving than the existing methods This method can be applied to numerous matrices for a finished dosage of selenium sulfide formulations.

Cryptosporidiosis is a parasitic disease caused by the protozoan Cryptosporidium in vertebrates. In livestock, especially ruminants, infants develop diarrheal syndromes. The infection is common worldwide , including Iran, where it is reported in several species. Morphological diagnosis of Cryptosporidium species is associated with many limitations and has no taxonomic value on its own, so the use of molecular methods can overcome these limitations to some extent. The present aims at microscopic, molecular and antigen detection and isolation of Cryptosporidium parvum parasites. Firstly, 300 samples were collected from different parts of Iran. Subsequently oocysts from feces were purified by the method of Casemore et al. using the flotation technique and stained by the modified Ziehl-Neelsen method (Henriksen method) and identified by diagnostic keys. ELISA test was also performed on the samples with results ranging from 1 to 4 positive. The results of our study show that, of the 300 cases tested for Cryptosporidium, 48 cases (16%) and 54 (18%) were positive in ELISA and PCR, respectively. Microscopic evaluation also mainly confirmed the ELISA results. These cases were collected in summer, autumn, and winter, with, more than 50% of the positive cases found among the samples collected in autumn. In addition, 54 positive cases were found by PCR test, which is 6 cases more than ELISA results. Finally, the results of PCR detection and ELISA were subjected to chi-square analysis, where no significant difference was found between the collected data (p=0.0587).

Shellfishes are a significant economic and nutritious seafood amongst people in different countries. Seafood products, particularly shellfish, are potential reservoirs of enteric viruses. This research investigated the incidence of rotavirus (RoV), norovirus (NoV) GI and GII, hepatitis A virus (HAV), and hepatitis E virus (HEV) in shellfish samples from the Persian Gulf, Iran. One hundred and fifty shellfish samples were collected. RNA extraction and cDNA synthesis were performed using commercial kits. The real-time polymerase chain reaction assessed the presence of enteric viruses in extracted cDNA samples. Thirty-two out of 150 (21.33%) shellfish samples were contaminated with enteric viruses. Prevalence rates of NoV GI, NoV GII, HAV, and RoV amongst shellfish samples were 8.00%, 11.33%, 1.33%, and 0.66%, respectively. There were no contaminated shellfish samples with HEV. Simultaneous prevalence of HAV and NoV GI, and HAV and NoV GII viruses were 0.66% and 0.66%, respectively. Examined viruses had a higher prevalence in shellfish samples collected in the winter season (P<0.05). Prevalence of HAV, RoV, NoV GI, and NoV GII amongst shellfish samples gathered in the winter season was 2.85%, 9.09%, 11.90%, and 20%, respectively. To the best of our knowledge, this was the first report of the incidence of enteric viruses, particularly HAV, NoV GI, NoV GII, and RoV, in shellfish samples from the Persian Gulf, Iran. Shellfish samples may serve as a potential source of enteric viruses for the human population. Therefore, routine viral assessments should be conducted. The consumption of fully cooked shellfish can significantly reduce the risk of HAV, RoV, NoV GI, and NoV GII infections. Furthermore, given the export value and importance of shellfish samples, their microbial quality and safety should be routinely monitored.

The biosynthesis of agglutinogenic and adsorbing groups A and B glycotopes of the erythrocyte's membrane is mediated by the activity of specific glycosyltransferases. This study aimed to assess the nature of the biosynthesis of A and B antigenic glycotopes, depending on the pH of the medium during the cultivation of erythrocytes, and the antigenic (transferase) characteristics of the donor serum of the other group. Monoclonal antibodies (Mabs) were obtained from IGBRL under Program IV of the International Workshop on Monoclonal Antibodies and Red Blood Cell Antigens. Biosynthesis was performed using erythrocytes, fresh serum, medium 199, and an antibiotic solution. The agglutinogenic characteristics of 11 out of 33 samples changed by the end of the cultivation period due to the acquisition of additional agglutinogen corresponding to the donor serum. None of the samples lost their inherent agglutinogen due to its absence in the donor serum. Four of six samples of O(I) erythrocytes acquired the ability to be agglutinated by anti-A reagents, especially by the polyclonal anti-A, and the manifestation of agglutination depended on the reaction time. Two of the three samples with initial A(II) agglutinogenic specificity added to the donor serum with Bc'+ characteristic of the erythrocytes acquired this characteristic. However, none of the five A(II)Ac'+ samples cultured in the serum of Ac'-O(I)Ac'-Bc'+ and O(I)Ac'-Bc'- donors lost their inherent earlier Ac'+ characteristic. The investigation of the inhibitory ability of alkaline and acidic glycoconjugates isolated from membranes revealed that alkaline Alp-00 and Alp-1 glycotopes isolated from glycolipids had the highest inhibitory activity, and the degree of inhibition of polyclonal anti-A antibodies was even higher than that of monovalent BRIC-131. This study showed the possibility of the biosynthesis of specific non-agglutinogenic A and B glycotopes under the influence of a different group's serum as a source of the corresponding transferase.