Archives of Razi Institute
Volume:79 Issue: 2, Mar-Apr 2024

Pro- or anti-inflammatory properties of cytokines in COVID-19: which offer better protection against diseaseI read the article by Al-Mquter et al. that compared circulating levels of 3 cytokines(IL-6, IL-25 and IL-35) amongst healthy controls(HC) and patients with COVID-19. They reported a significant increase in the serum concentration of these cytokines in patients versus HC(1). Their results confirmed again that assessment of cytokine profiles could be used as COVID-19 markers and may also provide practical implications for disease treatment. Nonetheless, understanding the role of cytokines in COVID-19 is complicated because of the complex nature of cytokine networks and the pathogenesis of COVID-19(2). IL-6 is one of the first cytokines produced during COVID-19 and increasing evidence suggests that its alteration may be associated with disease outcome. IL-6 is a pleotropic cytokine and has both pro- and anti- inflammatory effects. Therefore, it appears to operate in protective or pathological immunity to COVID-19. IL-6 can promote immune processes associated with resistance to different pathogens including COVID-19. It contributes to host defense via different pathways, including the stimulation of acute phase responses and hematopoiesis. Nonetheless, dysregulated and persistent synthesis of IL-6 can be resulted to the onset and development of pathological conditions. These different effector functions may be associated with different concentration of IL-6 that may result from different signaling pathwaysIL-6 signaling includes at least three distinct pathways: cis-signalling; trans-signalling and trans-presentation. It seems that IL-6 trans-signalling and trans-presentation are probably responsible for severe progressive COVID-19. These signaling pathways can trigger diffuse inflammation at various levels. Conversely, IL-6 cis-signalling mediates negative feedback mechanisms on proinflammatory cytokines, including suppression of their production, stimulation of their decoy receptors, and inhibition the maturation of Th17 cells. Circulatory IL-6 in COVID-19 patients reaches the peak level at advanced stages around two weeks of disease. Elevated IL-6 (e.g., > 100-120 pg/mL) levels in critical COVID-19 may reflect augmented IL-6 cis-signalling in an attempt to exert homeostatic functions, although there is no general consensus on this issue(3).

This study systematically investigated the role of Gummosin in improving memory in the scopolamine impaired memory model. Memory and learning are the most evolved and complex functions of the nervous system. Learning is the acquisition of new information that occurs as a change in behavior, and memory is the ability to store and retrieve learned information. In other words, memory is a combination of different processes of acquiring, consolidating, storing and recalling information. Memory consolidation and storage processes are the result of a series of time-dependent neurobiological events that occur after the initial formation of memory. In addition, this fluctuation in processes related to memory storage can fully occur shortly after the original learning experience. Memory is a direct result of learning due to storing and retrieving learned experiences and information. The results of our study showed that scopolamine causes impairment of memory, learning and synaptic plasticity, which was associated with changing the expression of various genes and reducing the number of hippocampal neurons. The disorders that occurred in scopolamine group rats confirm the model used in this study to induce memory and learning deficits, which, in line with previous studies, confirms the model used to induce Alzheimer's. The results of the behavioral tests of this study showed that, in accordance with previous works, scopolamine caused a significant increase in anxiety behaviors, which was associated with a decrease in the time spent in the central area compared to the control group, while donepezil injection led to a decrease in anxiety behaviors. It was possible to increase the time spent in the central area compared to the scopolamine group.

Gastric cancer, which is considered a main global health concern, is the sixth most frequent cancer and the second cancer-related death worldwide. This survey was designed to systematically review the anti-gastric cancer effect of all the organic and inorganic nanoparticles in in vitro, in vivo and clinical trials. The investigation followed the PRISMA guidelines and the findings were recorded in the CAMARADES-NC3Rs Preclinical Systematic Review and Meta-Analysis Facility (SyRF) database. We searched the publications in various English databases such as Scopus, Web of Science, EMBASE, PubMed and Google Scholar with no specified publication time frame to obtain papers regarding the anti-gastric cancer properties of nanoparticles. The following terms including: “nanoparticles”, “gastric cancer”, “anti-gastric cancer”, “metal nanoparticles”, “organic nanoparticles”, “inorganic nanoparticles “, in vitro”, clinical” and “in vivo” were applied in this study. Out of 11189 papers, 31 papers including 19 in vitro (45.5%), 3 in vivo (13.6%), 3 clinical trials (13.6%), and 6 in vitro / in vivo (27.3%) up to 2023 met the inclusion criteria for discussion in this systematic review. The most widely used nanoparticles were organic nanoparticles such as polylactic acid, poly lactic-co-glycolic acid (PLGA) (16, 80.0%), followed by inorganic nanoparticles such as silver nanoparticles (13, 41.9.0%). This review study showed the high anti-gastric cancer potential of a wide range of organic and non-organic NPs through their activity through some mechanisms such as induction of apoptosis, gene therapy, drug delivery, etc. However, more studies, especially in a clinical setting, are needed to confirm their anti-gastric effects and accurate mechanisms.

H. pylori is known to increase the risk of developing gastritis, peptic ulcer disease, gastric adenocarcinoma, and gastric lymphoma in adults, worldwide. This study aimed to determine the seroprevalence of H. pylori infection and risk factors associated with such infection among adult in Duhok province, Kurdistan Region, Iraq. This cross-sectional study was conducted among adult population in Duhok Province, Iraq. During the study period from 2018 to 2020, a total of 259 subjects aged more than 18 years who visited the general hospitals were included. H. pylori seropositivity rate was determined using un enzyme-linked immunosorbent assay (ELISA). A standardized questionnaire was administered to all study participants through face-to-face interview. The data obtained from H pylori IgG antibody were analyzed using the chi-square test. The prevalence of anti H. pylori IgG antibody positivity was present in 104 out259 (40.02%) of the adult. Among the studied variables, all the following risk factors were significantly associated with the presence of anti H. pylori IgG antibodies; male (p< 0.043), level of education (p<0.025), history of gastrointestinal diseases (p< 0.001), smoking status (p< 0.001) and a larger sibling number (p< 0.001). This study supports the hypothesis that H. pylori infection in adult is highly related to poor hygiene, smoking status, low level of education and crowded conditions. Therefore, it is crucial to implement effective strategies aimed at enhancing fundamental sanitary conditions, as well as improving educational level and socioeconomic status, in order to reduce the prevalence of H. pylori infection among adults in our region

In recent years, plague has re-emerged in several countries around the world and remains endemic in some regions. In a natural environment, and in contact with rodents and their fleas, stray carnivores are most at risk of catching the disease and maintaining the spread of the bacillus.The objective of this study is to demonstrate the presence or absence of Yersinia pestis in stray dogs and cats in the Algiers region by molecular methods and thus contribute their role in epidemiology of this disease. Molecular research of Yersinia pestis has also been conducted on fleas from these carnivores. Preliminary identification of ectoparasites to genus and species level was performed.Real-time polymerase chain reaction targeting Yersinia pestis pla gene was used to survey the plague agent in fleas and carnivores captured as stray animals in Algiers (Algeria). Positive qPCR results were tested by PCR sequencing using glpD gene.Among 327 fleas captured from 107 dogs and 365 fleas from 140 cats, prevalence of Ctenocephalides felis was higher in cats (86,96%), whereas that of Ctenocephalides canis and Xenopsylla cheopis was higher in dogs (90,57% and 92,63% respectively). While internal and external PCR positive controls were positive, none of the 107 dogs spleen and 140 cat spleens and none of the 256 analyzed fleas were positive for Y. pestis. These results suggest that stray cats and dogs are unlikely sources for plague in Algeria contrary to that has been reported in other plague-endemic countries. This observation illustrates that plague epidemiological chain varies from one region to another one.

Tegafur, gimeracil, and oteracil potassium are widely used pharmaceuticals to treat lung cancers of the gastrointestinal tract like the oral cavity, esophagus, colon and rectum, pancreas, and also non-small cell lung cancers. Literature review revealed that no study has yet offered a completely stability-demonstrating, validated liquid chromatography-mass spectrometric approach for the concurrent estimation of tegafur, gimeracil, and oteracil potassium along with all known degradation products. The simultaneous detection of tegafur, gimeracil, and oteracil potassium, and their forced degradation products characterization, necessitated the invention of a simple, faster, and less expensive method. The goal of the study was to follow ICH method validation standards to develop and validate a fast, easy, and rugged liquid chromatography-massspectrometry (LC-MS) technique for the concurrent estimation of Tegafur, gimeracil, and oteracil potassium in drug substance and finished dosage form according to ICH method validation guidelines. Tegafur, gimeracil, and oteracil potassium were examined on Waters HPLC Alliance system coupled to SCIEX QTRAP 5500 mass spectrometer endowed with an interface capable ofcarryout electrospray ionization. The tegafur, gimeracil, and oteracil peaks eluted at retention times of 2.338 min., 3.756 min., and 5.338 min. respectively. The limit of detection (LOD) values oftegafur, gimeracil, and oteracil were detected to be 0.6, 0.174, and 0.474 μg/mL, respectively however the results for quantification limit (LOQ) were calculated to be 2.0, 0.58, 1.58 µg/mLconcentration, respectively. Tegafur, gimeracil, and oteracil had linear ranges of 50-300 µg/ml, 14.5-87 µg/ml, and 39.5-237 µg/ml, respectively, with regression coefficients of 0.99956, 0.99986, and 0.999479. Accuracy values for tegafur, gimeracil, and oteracil in the ranges of 50%, 100% and 150% for each were respectively, were determined to be 99.9%, 99.9%, and 99.4%. The % RSD for six replicates was less than 2% for precision. According to ICH Q2 guidelines, this approach was effectively evaluated with LC-MS to validate the chemical structures of freshly created tegafur, gimeracil, and oteracil degradation products. An accurate and sensitive LC-MS technique was developed and validated for the concurrent quantification of tegafur, gimeracil, and oteracil potassium in drug material and medicinal dosage form.

Trichomonas vaginalis (T. vaginalis) and candida (C.) species are the most common causes of vaginal infections in reproductive-age women. T. vaginalis is a sexual protozoa parasite that causes trichomoniasis. Candida species are fungal and cause an infection in the female genital tract named candidiasis caused by several Candida species. Both microorganisms if not treated correctly may lead to many complications such as abortion, premature delivery, disorders of menstrual cycle, and infertility. The current study aimed to study the frequency of T. vaginalis and Candida species including C. albicans, C. krusei, and C. glabrata infections among females with vaginal infection in Duhok City, Kurdistan region, Iraq. About 250 vaginal swabs were collected from women that attended to the Vin Private Laboratory and 150 samples were collected from women that attended to the Arveen Private Laboratory have vaginal infections. Out of these 400 vaginal swabs samples, twenty-four samples were recorded positive for T. vaginalis by direct smear. One hundred ten samples were recorded positive for candidiasis by culturing on the Candida Chromagar. Three species of Candida were isolated including C. albicans, C. krusei, and C. glabrata, their prevalence was (60.9%., 28.25, 7.3%, and 3.6%) respectively. Vaginal infection was commonly found in the age group between 25-35 years (49.6%), followed by the group aged between 35-45 years (36.4%). Also, (3.2%) of samples were found to have a mixed infection with trichomoniasis and candidiasis. Because these two causative agents have many complications in women, therefore it is highly recommended a good control measurement to prevent or decrease vaginal infection such as health education, personal hygiene, and treatment of infected women.

The compound 1-methyltryptophan (1-MT) has been shown to act protectively in renal ischemia-reperfusion injury. Toll-like receptor 4 (TLR-4) signaling is also a regular process of epithelial to mesenchymal transition (EMT) that can after Ischemia-reperfusion injury (IRI) result in as an increase of renal fibrosis. EMT is associated with specific transcription factors – Snai1, Snai2, Zeb1 and Twist. 1-MT could regulate EMT and act as antifibrotic agent. This study aimed to investigate the effect of 1-MT on EMT transcription factors in tubular epithelial cells that underwent 30 min. Renal tubular epithelial cells (TECs) were isolated from Lewis rats using a standard protocol with Fe2O3 magnetic separation and selective media as previously mentioned. ischemia and 48 hours of reperfusion. Cells were cultivated and divided into 4 groups: C-TECs– control cells, IRI-TECs – IRI-induced TECs, D-IRI-TECs – IRI-induced TECs treated with 1-methyl-D-tryptophan, L-IRI-TECs – IRI-induced TECs treated with 1-methyl-L-tryptophan. IRI was induced in all groups for 30 min by mineral oil (except for C-TECs) followed by 48 hours of reperfusion. RNA and proteins were isolated from harvested cells. Using semi-quantitative PCR (RT-sqPCR) we assessed the relative mRNA expression of EMT transcription factors Snai1, Snai2, Zeb1, and Twist. Hereby we have shown, that the treatment of ischemia-induced TECs with both 1-MT isomers lowers the expression of EMT transcription factors Snai1 and Zeb1 that were increased by ischemia and reperfusion of TECs. This could act favorably in renal IRI decreasing EMT and renal fibrosis, therefore showing the potential of 1-MT as a part of therapy in renal transplantation aimed at renal ischemia-reperfusion injury.

Amoebiasis is an intestinal disease caused by a unicellular parasite called Entamoeba histolytica. Interleukin-35 (IL-35) is the most recent specific member in the IL-12 family, that plays a principal role in the inhibitory function of regulatory T cells (Tregs) to restrain inflammatory reactions. IL-25 from the IL-17 family, which is broadly released by Th2 cells and epithelial cells is a warning signal produced during cell or tissue injury for triggering immune cells. The present study was designed to determine the cytokine profile (IL-25 and IL-35) in patients with E. histolytica infection in Southern Iraq. This hospital-based study was carried out from August 2022 to May 2023. The study participants were patients suffering from E. histolytica infection referred to the Infectious Diseases Wards of the general hospitals in Thi-qar Province, Southern Iraq. At first, the E. histolytic amebiasis in patients was approved by the nested multiplex PCR. All collected sera were tested using the Human Interleukin 35 (Biotech, China, Cat.RD-IL35-Hu) and IL25 (Biotech, China, Cat.RD-IL25-Hu) ELISA Kits based on the producer’s instructions. A total of 80 patients including 50 patients with E. histolytica infection and 30 people in control group with no E. histolytica infection were included in the present study. The results showed a significant difference (p<0.001) in the serum level of IL-25 among patients with E. histolytica infection (4275.19 pg/mL) in comparison with people in the control group with no E. histolytica infection (2186 pg/mL). The statistical analysis showed that there is no significant difference in the serum level of IL-35 patients with E. histolytica infection in comparison with people in the control group with no E. histolytica infection. The findings of the present investigation revealed the high levels of IL-25 in patients with E. histolytica infection; indicating the important role of IL-25 in activating the immune system during intestinal inflammation. Therefore, this cytokine can be used as a diagnosis marker for E. histolytica infection.

Toxoplasmosis, which is created by the Toxoplasma gondii parasite is a parasitic, infectious disease. 10-hydroxy-2-decenoic acid (10-H2DA, queen bee acid (QBA), is one of the most prevalent fatty acid (>40%) presents in royal jelly. Studies reported various beneficial effects of 10-H2DA antitumor, anti-inflammatory, antiangiogenic, improving the immune system, and antimicrobial effects. This experimental survey aimed to evaluate the in vitro efficacy of QBA against tachyzoites and intracellular parasites of the T. gondii RH strain. Anti-Toxoplasma effects of QBA against tachyzoites were examined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay for 30, 60, 120, and 180 min. In addition, the effect of QBA on infection rate and intracellular parasites was studied. Real-time PCR was also applied to assess the expression level of the Caspase-3 gene. The best efficiency of QBA was reported at 100 and 50 µg/mL whereas all tachyzoites were diminished followed by 120- and 180-min treatment, respectively. We also found that the best repressing efficacy of QBA on the infection rate and the load of parasites into the Vero cells was indicated at 100 µg/mL (p<0.001): but, the combination of QBA (12.5 µg/mL) along with atovaqoune 30 µg/mL displayed the highest effect on the infection rate and the load of parasites into the Vero cells in the infected Vero cells. The expression level of the Caspase-3 gene was dose-dependently increased after exposure of tachyzoites to QBA; mainly at ½ IC50, and IC50 compared to the normal saline. The achieved findings exhibited the high in vitro potency of QBA especially in combination with atovaqoune against T. gondii RH strain tachyzoites. Although apoptosis induction can be suggested as one of the principle mechanisms; more studies are required to elucidate its accurate mechanisms as well as its efficacy and safety in animal models and clinical settings.

Anaplasmosis is a tick-borne disease with worldwide distribution, affecting ruminants, equines, carnivores, and humans. This study aimed to investigate Anaplasma phagocytophilum (A. phagocytophilum) in horses from Ardabil province and Anaplasma ovis (A. ovis) in small ruminants from East Azerbaijan province using the Nested PCR method. Blood samples were collected from the jugular vein of 100 healthy horses in the Ardabil province and 156 healthy sheep and goats (116 sheep and 40 goats) in the East Azerbaijan Province during the spring and summer seasons of 2016 in northwest Iran. The gathered blood samples were stored at -20°C until the molecular experiments were conducted. Nested PCR was employed to detect A. phagocytophilum in horses and A. ovis in small ruminants using extracted DNA and amplifying 16S rRNA and msp4 genes. The Chi-square test of independence was used to determine the relationship between Anaplasma spp., infection, and independent variables, including age, gender, animal species, and sampling location. None of the 100 samples collected from horses in the Ardabil province were positive for A. phagocytophilum. In the East Azerbaijan province, 11 out of the 156 (7.05%) blood samples collected from sheep and goats tested positive for A. ovis. In addition, A. ovis infection was not significantly related to the independent variables. Phylogenetic analysis showed that the sequence obtained in the present study (MH790273) has 100% homology with the sequence obtained from sheep infected with Anaplasma in Ahvaz (JQ621903.1). The study’s findings can aid in preventing and controlling anaplasmosis in farm animals in northwestern Iran.

Leishmania is the main cause of a serious public health problem called leishmaniasis in Iran. Pentavalent antimonial chemicals are usually used for leishmaniasis treatment. But this drugs have a number of negative side effects, including drug resistance, non-specificity, poor responsiveness, toxic effects, inconvenient injections, tissue damage and high cost. The aim of the present study was preparation and evaluation of the efficacy of green synthesized silver nanoparticles (Ag-NPs) against Leishmania infantum and Leishmania tropica in vitro. The MTT assay was used to assess the toxicity of Ag-NPs derived from ginger extract on macrophage cells. The apoptotic potential of promastigotes caused by Ag-NPs was evaluated using the flow cytometry method. According to our findings, proliferation of L. infantum and L. tropica, promastigotes are dramatically decreased by increasing doses of nanoparticles. The most effective doses of nanoparticle were 80 and 40 ppm after 48, and 72 hours of incubation respectively , while doses of 0.312 and 0.156 ppm after 24 and 48 hours of incubation had the least effect on the growth and activity of L. infantum and L. tropica promastigotes. For the promastigotes of L. infantum and L. tropica, the flow cytometry test revealed that Ag-NPs induced Programmed Cell Death (PCD) in promastigotes of L. infantum and L. tropica demonstrated 67.1% and 41.9% of apoptosis, respectively. The IC50 (inhibitory concentration) for NPs against L. infantum and L. tropica were 4.54 and 4.22 ppm, respectively based on MTT assay. The higher concentrations of NPs such as concentration 80 ppm, led to more lethality of promastigote. In conclusion, overall, Ag-NPs exhibited good in-vitro anti-leishmanial activity against L. infantum and L. tropica promastigotes.

In the era of free antibiotics used in animal production, the application of feed additives should be prioritized to improve poultry health and production. This study was conducted to evaluate the influences of garlic fermented by Lactobacillus spp. on the growth rate, intestinal microorganisms, and immune response of turkey broilers. A completely randomised design was used, involving 90 turkey broilers aged 1–56 days, with five treatments and three replicates per treatment. The birds were given feed and water ad libitum for entire the experiment period. The treatments included the supplement of aqueous extract from fermented garlic (FG) to drinking water. The results showed that broilers supplemented with 0.8% FG exhibited the largest final body weight (1,158 g/bird), body weight gain (19.64 g/bird/day), and significantly improved feed conversion ratio (1.962) while decreased the feed intake of turkey broilers from to 1–56 days. The immune organ indices including the spleen, thymus, and bursa of Fabricius indices were increased in the treatment of 0.6% FG (P<0.05) while antibody titres (at 28 and 42 days of age) were improved in the 0.6% and 0.8% FG treatment (P<0.05). Clostridium perfringens and Salmonella spp. were not detected in the intestines of these birds, while the amount of Escherichia coli was reduced (P<0.05) and Lactobacillus spp. increased (P>0.05) without a significant effect. It can be concluded that supplementation with 0.8% FG improved growth performance, and 0.6% FG may enhance the immunity of turkeys. The use of 0.6 and 0.8% FG could be widely applied for poultry production.

Diphtheria is a re-emerging disease with changing epidemiology. Recognising diphtheria's clinical profile, patterns of morbidity and mortality, and level of immunisation is very essential. This study was planned to study the clinic-epidemiological profile of patients with Diphtheria at Infectious disease Hospital, New Delhi. The present study was a cross-sectional study conducted at Infectious Disease Hospital, Delhi included 94 patients who were diagnosed with Diphtheria were included in the study. The study tool was a questionnaire that had questions eliciting details such as socio-demography, clinical history, examination findings, vaccination history and clinical history. The case fatality rate was calculated. Means and proportions were calculated. A p value of <0.05 was considered significant. The mean (SD) of the age of the participants was 9(4.4). Laryngeal involvement was found in 10(11%). Complete diphtheria vaccination doses were taken by 6(6.4%). The case fatality rate was 13%. Complications such as Neuropathy were found in 21(22%), cardiac problems in 12(13%) and respiratory problems in13(14%). Longer duration of illness, delayed presentation and complications were associated with poor clinical outcomes. (P-value <0.05).The majority of diphtheria cases belonged to the age group of 6-10 years and lower socio-economic group. Most of them had partial vaccination against diphtheria. Around one-third of patients had complications related to diphtheria and the case fatality rate was high. Poor clinical outcomes were linked to longer disease duration, delayed presentation, and complications. Key preventive measures for controlling the deadly diphtheria disease include specialised immunisation programmes for poor immunisation coverage areas and early detection and treatment of suspected cases.

Human papillomavirus (HPV) has been established as a causative agent in the development of oral squamous cell carcinoma (OSCC). Specifically, HPV types 16 and 18 are known to be prevalent in oral cancers. This cross-sectional study aimed to determine the prevalence of HPV types 16 and 18 in OSCC cases in Qazvin province, Iran. Thirty-eight paraffin-embedded samples of OSCC was selected, and DNA extraction was performed using the Roche High Pure FFPE DNA isolation kit. The quality of the extracted DNA was assessed through PCR amplification of the human β-Globin gene. HPV detection was carried out using SYBR green-based real-time PCR with GP5+ and GP6+ primers targeting the L1 region of HPV. HPV genotyping was conducted on positive samples using specific primers. Statistical analysis was done between HPV infection in OSCC and age, sex, and anatomical location (p < 0.05 considered significant). This study analyzed 38 biopsy specimens obtained from male and female OSCC patients, with an average age of 64. Among these samples, 13 tested positive for HPV, resulting in a prevalence rate of 34.2%. The age group with the highest HPV infection rate was 61–70 years (10.5%). Notably, HPV type 16 was detected in 21.0% of the samples, HPV type 18 in 10.5%, and other viral subtypes in 2.6%. No statistically significant correlation was found between HPV prevalence and gender or age. The findings indicate that 34.2% of OSCC samples in the Qazvin province harbor HPV, with types 16 and 18 being the most common in tumors affecting the tongue. Additionally, no association was observed between HPV infection and age or gender. To address HPV as a risk factor for OSCC, public health initiatives such as vaccination, awareness campaigns, and accessible healthcare services should be implemented. They are, furthermore, incorporating HPV DNA testing into practice.

MicroRNAs (miRNAs) are a class of small non-coding RNAs of 21–25 nucleotides in length and display an essential role in regulating cancer initiation, development, and progression. Breast cancer (BC) is the most commonly detected malignancy in women, and it is one of the main motives of death worldwide. In this study, the impacts of microRNA-451a-5p and miR-34a-5p (tumor suppressors), individually and combined, transfections were conducted on the apoptosis, proliferation, and migration of breast cancer cells in vitro. For carrying out this research, malignant breast cancer cells (MDA-MB-231) were transfected with miR-451a-5p and miR-34a-5p mimics. Then, the cytotoxicity, apoptosis, proliferation, migration and, the protein and gene expression of caspase-3, caspase-8, MMP9, ROCK, vimentin, c-Myc of the cancer cells were assessed by MTT, flow cytometry, q-RT-PCR (expression levels of caspase-3, caspase-8, MMP9, ROCK, vimentin, c-Myc genes), wound healing, and western blot assays. The resluts indicated that the miR-34a-5p and miR-451a-5p could additionally induce apoptosis and cell cycle arrest in the sub-G1-phase, repress the proliferation and migration in the breast cancer cells, and could also decrease β- catenin, ERK/P-ERK protein expressions. The present data documented that restoring tumor suppressor miR-451/miR-34 in vitro strongly induced the programmed cell death and obviously inhibited the cell proliferation and migration in human breast cancer cells. Taken to gather, miR-451a and miR-34a have a considerable role in breast cancer cell proliferation and migration ability via Wnt/β-catenin and ERK/P-ERK signaling pathways. Therefore, the simultaneous restoration of the presented tumor suppressor miRNAs may be proposed as a valuable and potential therapeutic strategy in breast cancer treatment. However, further study should be meaningful.

Galliformes have a global distribution and are bred by humans as domestic animals or game birds. The world's poultry population is increasing due to high consumer demand for poultry products. Due to the increasing growth of Galliformes breeding, this study aimed to investigate gastrointestinal parasites in the feces of some species of Galliformes.Galliformes have a global distribution and are bred by humans as domestic animals or game birds. The world's poultry population is increasing due to high consumer demand for poultry products. Due to the increasing growth of Galliformes breeding, this study aimed to investigate gastrointestinal parasites in the feces of some species of Galliformes. In this study, 100 distinct fecal samples were collected from seven different Galliformes species, including rural chickens, peacocks, partridges, pheasants, turkeys, quails, and guinea fowls. Sampling of each bird was done separately. These birds were kept in cages. To identify gastrointestinal parasites, the samples were evaluated using the Clayton-Lane method, modified Ziehl-Neelsen, and Trichrome staining methods. Out of 100 birds sampled, 53% of birds were positive for gastrointestinal parasites. 30% of birds were infected with Eimeria spp., 18% of birds were infected with nematodes, and 5% of birds were infected with Giardia spp.. The highest rate of parasitic infestation in Galliformes was related to protozoan infections and the highest rate of protozoan infection was related to Eimeria (30%). Complex infestation was not detected. Considering that most parasitic infections have been detected in apparently healthy birds, it is recommended to observe hygiene and to regularly disinfect cages and reduce the density of birds in the nests. Due to the zoonotic potential of Giardia and due to the contamination of some birds with this parasite, this issue should be considered by public health officials, bird sellers' markets, owners, and breeders of these birds.

Canine Parvovirus 2 is one of the main etiologies of viral gastroenteritis in dogs worldwide. This disease is characterized mainly by the presence of diarrhea, abdominal pain, vomiting, anorexia and dehydration. This virus is responsible for a high mortality and morbidity rate in dogs younger than 3 months and those incomplete or not vaccinated. Monitoring of viral variants in our region has shown that in the last 7 years, variant 2c has been circulating exclusively, which is unusual if we consider that in the rest of the world at least 2 variants co-circulate among dog populations. In Mexico, there are no studies that report genomic sequences of CPV-2 which are relevant for population comparisons at the genetic level; thus, the purpose of the present study was to sequence genomes associated with CPV-2c. To meet this objective, stool samples were collected from dogs with suspected CPV-2 infection; five positive cases diagnosed by lateral flow testing and PCR were selected for viral genome sequencing. Comparative analyses showed that the genome sequences obtained were >99% homologous to those reported for CPV-2 in GenBank. On the other hand, 52 nucleotide mutations were identified in the vp1/vp2 gene of which 3 impacted the amino acid transition (T226S, F267Y, A440T). Phylogenetic analysis of the vp1/vp2 gene showed that the five sequences cluster in a clade called "III", which is related to sequences from USA and Uruguay. To our knowledge, this is the first report of genomic sequences associated with CPV-2 in Mexico, which is of great relevance for the epidemiological-molecular understanding and evolution of the virus.

Obesity has been an important health concern over a decade, causing serious health issues worldwide. Treatments available for obesity include FDA approved drugs like Lorcaserin, Orlistat, Bupropion, combinations of Phentermine and Topiramate, Sibutramine but these have adverse effects on health. To address the issue the current study was performed on evaluating the anti-obesity potential of Phyllanthus fraternus leaves. These leaves are a rich source of different phytochemicals like alkaloids, saponins, terpenoids, tannins etc., and the plant has showed to exhibit medicinal properties hence it can be used in the treatment of obesity disorder. The crude extract of plants was prepared in three different solvents i.e., methanol, hydro alcohol and isopropyl alcohol. Determination of lipid inhibition was done using lipase inhibition assay and Amylase assay was carried out to determine if the plant extract possessed anti-diabetic properties. An Oil red staining was carried out to determine lipid accumulation in which the cells were incubated with plant extract for 48 hrs. In order to determine if the plant extract was toxic to 3T3 cells MTT assay was carried out to access cell viability. Through lipase inhibition assay we depicted potent anti-obesity property, isopropyl alcohol extract exhibited 67.45% inhibition at the concentration of 500µg/ml. Methanol extract showed highest percent of α amylase inhibition i.e., 90.03% at a concentration of 1000µg/ml. MTT assay concluded that the plant extracts were not cytotoxic to the cells at a concentration range between 20µg/ml to 100µg/ml the percentage of viable cells was 98% to 63%. The plant extract successfully could be used to treat obesity. The results obtained from the current study revealed that the plant exhibits potent anti-obesity properties.

Due to reduced levels of antibody provided by vaccines or the emergence of variants COVID-19 infections have re-emerged in many countries. A considerable supplementary intervention is recommended including an additional dose of vaccination. The present study aimed to investigate the predictors of the willingness to receivethe third dose of COVID-19 vaccine based on the health belief model (HBM) constructs among people referring to health centers in South Khorasan province. In this cross-sectional-analytical study, 283 people over 18 years in South Khorasan province were randomly selected by multi-stage cluster sampling method. The data were collected using a researcher-made questionnaire including demographic information and health status, knowledge about COVID-19 and the third dose of the vaccine, as well as a questionnaire on based HBM constructs through self-report. Finally, the data were analyzed using Chi-square, and multivariate logistic regression. The mean age of the study participants was 36.57±11.56 with range of 18-66 years. The results of multivariate logistic regression analysis showed that gender (P=0.012), marital status(P=0.038), occupation (P=0.013), and constructs of perceived severity (P=0.005), and cues to action (P=0.018), had a significant direct effect and the construct of perceived barriers (P=0.010) had a significant inverse effect on the willingness to acceptance the third dose of the vaccine. Also, the predictor variables explained about 67.7% of the tendency to receive the third dose of the vaccine. The current study suggests that enhancing the perceived severity of Covid-19, along with healthcareproviders recommendation to receive the vaccine and reducing perceived barriers, can effectivelyencourage individuals to receive the third dose of the Covid-19 vaccine. These findings can be utilized to develop interventions aimed at promoting the uptake of the booster dose of the Covid-19 vaccine.

Since about 100 years ago, the best and main treatment for millions of global snakebite victims has been the use of polyclonal antivenoms. However, common antivenoms need continuous improvement to reduce rare, their side effects and get faster performance. In this study, by papain digestion of anti-cobra venom plasma and subsequently multi-step purification and optimization, including, ammonium sulfate precipitation and subsequently DEAE-cellulose column chromatography, Fab antivenom was produced. Then, by SDS-PAGE method, and double immunodifusion (Ouchterlony) tests the existence of the corresponding Fab fragment antibody was seen and confirmed. Also, in potency test in NIH laboratory mice, it was found that each milliliter of the new Fab antivenom was able to neutralize 624 micrograms, (80LD50) of cobra venom, which is about 15% more efficient than the primary plasma in the same concentration and also, 1.57 times more than the cobra antivenom which found in commercial hexavalent antivenom of Razi Institute respectively. According to the results, it seems that this new Fab antivenom can be used as a new candidate for the treatment of cobra snake bite victims.  

Obesity is the result of positive energy balance in which various hormones and neurotransmitters are involved. Using Dietary supplements is a common and popular method to lose weight. Medicinal plants with specific effects on metabolizing enzymes, blocking adipogenesis and improving energy metabolism can be a suitable alternative to these supplements. In this study, the role of the hydro-alcoholic extract of Dactylorhiza maculate (Salep) plant on obesity and its related hormones and antioxidants was investigated. Forty male Wistar rats were divided into five groups: Control, Sham, and Salep extract groups (three doses): 80, 160, and 320 mg/kg. The extract was fed by gavage for 29 days. After the 29th day, blood and tissue samples were taken. Rats’ ELISA kits were used to measure adiponectin, obestatin, resistin, orexin-A, insulin, epinephrine, Agouti Related Neuropeptide (AgRP), omentin, chemerin, amylin, neuropeptide-Y (NPY), and ghrelin. In addition, we measured leptin, cholecystokinin, antioxidants, and lipid profile factors. Evaluation of weight changes showed that Salep extract helped the animals to lose weight significantly in 160 and 320 mg/kg Salep groups. Leptin, adiponectin, AgRP, obestatin, CCK, chemerin, adiponectin, and total antioxidant showed a significant increase compared to the control group. In contrast, ghrelin, omentin, resistin, NPY, amylin, orexin-A, epinephrine, and MDA decreased in the Salep groups. Lipid profile was also affected by the extract. These findings suggest that the Salep extract prevents appetite, reduces ghrelin, and affects digestive factors; the Salep extract can change the secretory factors of adipose tissue and lipid profile, and ultimately help to lose weight.

In Ayurveda, Vitex negundo (VN) is used as a drug to manage pain, inflammation, problems related to polycystic ovary disease and the menstrual cycle. The bioactive compounds isolated from this plant exhibit anti-inflammatory, anti-oxidant, anti-cancer and microbicidal properties. The shrub Vitex negundo is known for its role in the modulation of cellular events like apoptosis and cell cycle. There is still a scarcity of data in the literature on the cytotoxic activity of Vitex negundo extracts on ovarian cancer. Hence, in this study, the phytochemical composition, anti-oxidant and anti-cancer activities of leaf extracts were evaluated. The chloroform and methanol fractions exhibited higher phenolic content (161.04 ± 0.02 mg/g GAE and 152.56 ± 0.05 mg/g GAE respectively) as compared to other fractions. The aqueous and petroleum ether fractions exhibited higher flavonoid content (515.27 ± 0.28 mg/g QE and 211.82 ± 0.05 mg/g QE respectively). The acetone and methanol extracts showed significant anti-oxidant capacities. Both leaf extracts of Vitex negundo inhibited PA1 cancer cell growth in a dose-dependent manner with IC50 values of 88.01 ± 3.14 and 112.30 ± 1.93 μg/ml respectively, as compared to the standard drug Doxorubicin with IC50 value 2.91 μg/ml (p < 0.05, One - Way ANOVA). The Gas Chromatography - Mass Spectroscopy (GC-MS) analysis allowed us to identify twenty- five bioactive compounds in acetone extract and twenty- two compounds in methanol extract. Therefore, further studies should focus on the isolation of novel compounds, which are more effective and less toxic, and constitute interesting substitutes for the development of anti-cancer drugs.

Newcastle disease is a severe viral threat to the global poultry industry due to its high prevalence and rapid transmission. Evaluating vaccination timing and effectiveness is crucial, often accomplished through the hemagglutination inhibition (HI) assay. This test relies on the virus's agglutination ability in certain animals, utilizing various inactivated antigens. Our study aimed to assess multiple Newcastle viral antigens ( LaSota, clone, thermo-resistant strain, B1, and V4 ) inactivated by binary ethylene amine (BEA) and formalin, seeking the best antigen and inactivator for the HI assay. We prepared the different ND antigens include; LaSota, Clone, thermo resistant, B1, V4 and the mixture of the antigens then inactivated them using BEA and formalin. The hemagglutination (HA) assay determined mean titers, comparing BEA and formalin inactivation. These antigens were also subjected to the HI test using 112 serum samples from different commercial poultry flocks to assess their performance. BEA-inactivated antigens exhibited significantly higher mean titers in the HA assay than formalin-inactivated antigens. In the evaluation of different antigens in the HI test, the mean titer of antigen B1 followed by clone and LaSota displayed a higher mean titer than others. In conclusion, this study recommends using Hitchner pathotype antigens, specifically the B1 vaccine, for Newcastle disease HI testing. BEA is the preferred inactivator, preserving antigen structure particularly the structure of hemagglutinin antigen while minimizing risks. These findings can enhance serological testing accuracy, contributing to more effective disease control and prevention in the poultry industry.