Archives of Razi Institute
Volume:78 Issue: 5, Sep-Oct 2023

Most chemicals expressed in mammalian cells have complex delivery and transport mechanisms to get to the right intracellular sites. One of these mechanisms transports most transmembrane proteins, as well as almost all secreted proteins, from the endoplasmic reticulum, where they are formed, to their final location. Nearly all eukaryotic cells have a membrane trafficking mechanism that is both a prominent and critical component. This system, which consists of dynamically coupled compartments, supports the export and uptake of extracellular material, remodeling and signaling at the cellular interface, intracellular alignment, and maintenance of internal compartmentalization (organelles). In animal cells, this system enables both regular cellular activities and specialized tasks, such as neuronal transmission and hormone control. Human diseases, including neurodegenerative diseases, such as Alzheimer's disease, heart disease, and cancer, are associated with the dysfunction or dysregulation of the membrane trafficking system. Treatment and cure of human diseases depends on understanding the cellular and molecular principles underlying membrane trafficking pathways. A single gene mutation or mutations that result in impaired membrane trafficking cause a range of clinical disorders that are the result of changes in cellular homeostasis. Other eukaryotic organisms with significant economic and agricultural value, such as plants and fungi, also depend on the membrane trafficking system for their survival. In this review, we focused on the major human diseases associated with the process of membrane trafficking, providing a broad overview of membrane trafficking.

The COVID-19 disease emerged in Wuhan, China, in December 2019 and quickly became a global health threat. Around 6,947,192 people have been killed around the world so far, including 146,292 in Iran. In addition to the definitive diagnosis of the disease by RT-PCR, immunological and serological tests can check the anti-SARS-CoV-2 N protein antibody titer in people at different stages of infection with acceptable sensitivity and specificity. The serological examination is an effective and efficient method for determining the prevalence of the disease, especially when asymptomatic cases are present or the diagnosis of symptomatic cases is incomplete. The study examined the seroprevalence of COVID-19 at the Razi Vaccine and Serum Research Institute (RVSRI) and the Agricultural Research, Education, and Extension Organization (AREEO). A total of 493 blood samples were collected from volunteers in June 2020 in AREEO, and 380 samples were collected in June and July 2020 in RVSRI. The total number of volunteers from both organizations was 873. Standard ELISA kits were used to measure IgG and IgM antibodies against SARS-CoV-2. A statistical analysis of the obtained data was conducted using SPSS (version 22.0). Among the total 873 volunteers examined in RVSRI and AREEO, 10.5% had elevated serum titers either for IgM or IgG, 3.55% of whom were women and 6.95% were men. Generally, 8.8% of people tested positive for IgM, which showed a recent infection with COVID-19 in people at that time and partially indicated the start of a new wave of COVID-19. In RVSRI, 3.42% of people with positive IgM titers (positive or negative IgG titers) were women and 5.53% were men. In AREEO, 3.02% were women and 5.72% were men. The seroprevalence rate of COVID-19 in RVSRI was 11.6%, 4.2% of which were women and 7.35% were men, with no significant difference between women and men. The COVID-19 seroprevalence in AREEO was 9.7%, 3.22% of which were women and 6.5% were men, with no significant difference between women and men.

Newcastle disease (ND) is a highly contagious viral infection affecting poultry production in many countries. Strict biosecurity and the administration of live attenuated vaccines against the ND virus (NDV) are the main implements of controlling programs. This study evaluated the efficacy and potency of the Razi Clone12IR Newcastle vaccine in specific pathogen-free (SPF) chickens. Chickens were vaccinated with either the Razi Clone12IR vaccine (group A1, n=20) or an imported Clone vaccine (B1, n=20) in the first week of life and boosted in the second week via eye drop, while negative control chickens received PBS (C1, n=20). Half of the birds in each group were challenged with the virulent NDV strain in the third post-vaccination week (A2, B2, and C2 groups). Specific antibody responses were determined in the collected sera by the hemagglutination inhibition (HI) assay for up to eight weeks. Cell-mediated immunity (CMI) was determined by the lymphocyte proliferation assay three and six weeks after the second vaccination. Sections of the tissues and organs, including the trachea, lungs, cecal tonsils, spleen, the bursa of Fabricius, liver, and small intestine, were subjected to histopathology. The immunized groups A1 and B1 showed significantly higher HI antibody titers before the challenge than the control group. In addition, lymphocyte proliferation responses significantly increased in the peripheral blood of the vaccinated groups. After the challenge, the A2 and B2 groups conferred good protection and drastically reduced virus shedding. No main lesions were noted in the tissues or organs of the vaccinated group in histopathology. In a few cases, mild microscopic lesions were observed, including the infiltration of inflammatory cells, which was related to the effect of the vaccine virus. These results indicate that the Razi Clone12IR vaccine is safe and can be an efficient tool for NDV infections by inducing protective humoral and CMI responses.

In the last decades, numerous studies have focused on the search for new agents to suppress the growth of cancer cells. In this study, we investigated the effect of two novel synthetic coumarin derivatives, namely 2-amino-4-(4-(2-hydroxyethoxy)-3-methoxyphenyl)-5-oxo-4H,5H-pyrano[3,2-c]coumarin-3-carbonitrile and 2-amino-4-(4-hydroxyphenyl)-5-oxo-4H,5H-pyrano[3,2-c]coumarin-3-carbonitrile, on the induction of apoptosis in breast cancer in a mouse model. Breast cancer was induced in BALB/c mice, which were randomly divided into six groups and then underwent the experiment. The groups and treatments included A1: coumarin A with a low dose (10 µm), A2: coumarin A with a high dose (1 mM), B1: coumarin B with a low dose (10 µm), B2: coumarin B with a high dose (1 mM), D: doxorubicin, and C: cancer control/ treatment with normal saline. The samples underwent treatments for 5 weeks. Animals were euthanized, and tissue samples, including the lung, liver, and tumor mass, were collected for histopathological examination. In addition, quantitative real-time polymerase chain reaction (qRT-PCR) was performed to determine some apoptotic markers, such as BCL-2, caspase-9, COX-2, and c-Myc. The qRT-PCR presented that both coumarin compounds could significantly alter the expression levels of BCL-2, caspase-9, COX-2, and c-Myc. Consistent with these results, histopathological observations showed a significant reduction in pathological lesions and severity of malignancy of the tumor mass, as well as a decrease in microscopic metastases in the lung and liver. This suggests that the present new coumarin compounds may induce apoptosis in breast cancer cells by altering some apoptosis-related genes that may play a chemotherapeutic role in breast cancer therapy in the future.

Consumption of contaminated water and foods by Salmonella Typhi cause the most common enteric disease known as Typhoid fever in both humans and animals. Despite the existence of various vaccines but infectious diseases remain a major cause of mortality worldwide. Nowadays, in-silico tools design a reliable and stable vaccine to combat such infections. The study aimed to design and evaluate a multi-epitope vaccine based on the outer-membrane proteins of Salmonella Typhi. B-cells and T-cells epitopes were predicted. Predicted epitopes were connected by AAY, KK, and GPGPG linkers. Heparin-Binding Hemagglutinin Adhesin (HBHA) has been attached to the N-terminal of the final vaccine as a potent immune adjuvant. Epitope’s antigenicity, allergenicity, immunogenicity, and physicochemical characteristics were defined using in-silico tools. Molecular docking of vaccine-TLR4 was done. ∆G of vaccine-TLR4 is -3.91×104 Kcal mol-1 with 1.93 RMSD. The results indicated protein was stable and non-allergen. In conclusion, the multi-epitope vaccine base on outer membrane proteins of the Salmonella Typhi bacterium might be considered to combat typhoid fever.

Kidneys are critical in the clearance and maintenance of active metabolites. One of the medical properties of Salep is treating bladder and kidney inflammation. Due to the widespread use of Salep in traditional medicine and the food industry, and since the effects of Salep on kidney function have not been studied, the present study aimed to investigate the impact of Salep on kidney function. In this experimental study, 48 male rats were divided randomly into six groups as control, sham, and four experimental groups receiving different doses of Salep intraperitoneally (80, 160, 320, and 640 mg/kg). On day 29, after weighing the animals, blood samples were taken from the heart, and serum blood urea nitrogen (BUN), uric acid, and creatinine were analyzed and compared in different groups. All the animal’s kidneys were exposed after dissection, and tissue sections were prepared for histopathological evaluation. From day 28 to 29, rats were kept in metabolic cages to collect urine samples and measure water intake and urine volume. The serum concentration of BUN and uric acid in the groups receiving Salep at all doses decreased non-significantly compared to the control group. Furthermore, a significant reduction was seen in creatinine serum levels in groups receiving 320 and 640 mg/kg of Salep extract (P<0.05). No evidence of damage to renal tissue was observed in this study. In conclusion, Salep could decrease serum BUN, uric acid, and creatinine levels due to its antioxidant properties and had no devastating effect on kidneys.

The yellow digger scorpion, Scorpio maurus, is a medically important scorpion for which little is known about its genetic diversity. Polymerase chain reaction products of 16srRNA gene fragments were generated from scorpion specimens named SmKh1 and SmKh2. These sequences showed high similarity with the only partial sequence of S. maurus isolate SCA1 large subunit ribosomal RNA gene available in the Genbank database. The drawing of the phylogeny tree showed two clusters, A and B. The two specimens (SmKh1 and SmKh2), which are placed in sub-cluster A2, were provided from Behbahan, Iran, and they have the closest relationship with the only sequence of S. maurus (MW281771), which is also collected from Behbahan. It is noteworthy that the two sequences obtained from S. maurus scorpions recorded from Miandoab (MK170444) and Mahabad (KU705354), which are in sub-cluster A1, are more similar to the scorpions isolated from the Mediterranean basin than those collected from Behbahan. This issue is probably due to the fact that patterns of genetic diversity are a reflection of variation in gene flow, which is also influenced by factors such as territorial barriers and geographical distances. We conclude that the scorpions of this study accompanied by similar scorpions in the Mediterranean basin, belong to the same species despite the insignificant differences.

Scabies is considered an external parasite notorious for its high prevalence causing severe and contagious skin lesions in humans and animals worldwide. This study has introduced a medicine to treat dogs infested with scabies (variants of Demodex, Sarcoptes, Psoroptes, Otodectes, etc.). The present study offers a no-side-effect herbal formulation to treat dogs infested with scabies. Unlike oral and injectable medicines, which take the form of an ointment and are topically applied on-site, this medicinal formulation can be easily used without concerns over its side effects or consumption dosages. This medicinal formulation requires no skin rinsing due to its herbal and high skin absorption properties, as recovery may take less than a month with a maximum of two times of application. To carry out the experiment, 25 sick dogs with various breeds and ages suspected of scabies were gathered. Following accurate morphological examinations of all the samples, a deep skin chip of the lesion site was provided, which was examined by a microscope. Then, 13 dogs (Mix, Terrier, Pug, Husky, Spitz) were infested with Demodex scabies and 12 dogs (Pittbull, Mix, Shih Tzu, Terrier, Boxer, Setter) with Sarcoptic scabies. The prepared product was topically administered at a constant 2% dosage to the bodies of all the samples. To prepare the ointment, 1 g of Borax (Na₂B₄O₇·10H₂O) was first dissolved in 35 g deionized water and heated to 70°C. Then, 45 g of liquid paraffin (CnH2n+2) was mixed with 1 g of Carvacrol (C10H14O) and 1 g of geranium (C10H18O) and stirred well to become a phase. Later, 17 g of the melted beeswax (C15H31COOC30H61) was added to the liquid paraffin compound. In the end, the aqueous phase was added to the oil phase, and the mixture process immediately began in one direction with a glass stirrer and continued until the product cooled down. Essential oils (EO) was obtained by steam distillation of fresh Thyme and Rose-Acented Geranium in a stainless steel distillation apparatus (alembic) for 3 h. The main components of the essential oils used in the formulation were performed using a Hewlett-Packard GC system interfaced with a mass spectrometer equipped with an HP5-MS capillary column (30 m, 0.32 mm, 0.25 µm film thicknesses). For GC–MS detection, electron ionization with ionization energy of 70 eV was used. To examine the presence of scabies, weekly skin sampling was performed, and the treatment continued until 30 days, when no skin chip of the scabies was noted. The findings revealed that the formulation developed no side effects and removed the daily use, as it could be administered once or twice a week. Also, complete recovery of scabies in all the breeds was found to be less than a month at most. This medicinal formulationcan be easily used without concerns over its side effects or consumption dosages. This study introduced a herbal formulation with effective herbal ingredients without any side effects to treat the sarcoptes and demodex parasites; unlike other chemical compounds, this medicinal formulation has no side effects, while some other formulations could develop side effects.

Foot and mouth diseases are among the important threats in the animal husbandry industry which lead to huge economic losses. In this regard, the current project aimed to inhibit the VP1 protein of foot and mouth disease viruses using specific peptides. For this purpose, a wide range of potential antiviral peptides were collected from the database. Physicochemical properties, hydrophobicity/hydrophilicity, and solubility properties of potential antiviral peptides were investigated using reliable servers. Afterward, the tertiary structures of the selected peptides along with the VP1 protein were modeled by the I-TASSER server. Moreover, interactions between VP1 protein and selected antiviral peptides were investigated using the ClusPro 2.0 server. Finally, the outputs of molecular docking were assessed by LigPlot+ and visualized by PyMol software. The results revealed that Dermaseptin-3, Ginkbilobin, Circulin-F, Maximin1, Cycloviolin-A, Cycloviolin-D, Circulin-C, Cycloviolin-C, and Antihypertensive protein BDS-1 peptides with a hydrophobicity value of > 30 were soluble with positive instability index and positive net charge. Moreover, the results of the molecular docking process demonstrated that Dermaseptin-3 and Ginkbilobin peptides could strongly inhibit the VP1 protein using 10 hydrogen bonds. Therefore, these two peptides, which had the most hydrogen bonds, were introduced as the best anti-foot and mouth disease virus peptides to apply.

Epstein-Barr virus (EBV), one of the most significant causes of lymphoid and epithelial cancers, has been linked to oral carcinogenesis; however, this etiological association remains controversial. To investigate this association, the present study aimed to determine the prevalence of EBV in cancerous and non-cancerous oral tissues from Ahvaz, Iran. In total, 164 blocks of formalin-fixed paraffin-embedded tissues from oral squamous cell carcinoma (OSCC), including 76 tongue squamous cell carcinomas and 88 non-cancerous tongue tissues, were collected from Ahvaz Imam Khomeini Hospital, Ahvaz, Iran, from December 2014 to March 2019, for this case-control study. The tissues were cut into 15-μm-thick sections, and DNA was extracted using a solution of Phenol, Chloroform, and Isoamyl Alcohol. The EBV detection and typing were performed using nested polymerase chain reaction. The EBV was detected in 9 (5.48%) out of the 164 samples studied, including 4 (5.26%) of the 76 SCC cases and 5 (5.68%) of the 88 samples in the control group (P>0.05). The EBV was positive in 2.40% of the 83 male and 8.6% of the 81 female samples (P>0.05). In terms of the histological grades of the case group, 3 (3/57) and 1 (1/13) of the EBV-positive samples were well and moderately differentiated, respectively (P>0.05). For EBV typing, the 9 EBV-positive samples were tested, and it was found that 2 and 7 of the cases were EBV type I and II, respectively. Results of the current study demonstrated the low frequency of EBV in Iranian patients with OSCC, with EBV type II predominating. Further studies are required to clarify the association between EBV and OSCC.

The Iranian Echis Carinatus (IEC) venom is an exclusive natural source of bio-substances for a wide range of purposes in the blood coagulation cascade. The present study for the first time was aimed to assess novel pro-coagulant, anti-coagulant and anti-platelet proteins, named EC1.5 (a), EC5.1 (b) and EC4 (a) from Iranian Echis Carinatus (IEC) venom.These peptides were purified by multi-step chromatography methods. Hematological properties were measured using activated clotting tests, platelet aggregation studies, and hemorrhage assessment. Subsequently, these proteins were identified through both their intact molecular mass and peptide mass fingerprint (PMF) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Multiple sequence alignments were performed by ClustalW, Bioedit software. Molegro Data Modeller (MDM) 3.0 software was used to predict the putative tertiary structure of proteins.EC1.5 (a), a single-band protein with a molecular mass of 66 and 55 kDa, was observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a reduced and non-reduced state, respectively. Based on the Mascot results, we considered that EC1.5 (a) is a metalloproteinase of group ΙΙ which exhibited potent pro-coagulant activity. It is predicted that the EC1.5 (a) with hemorrhagic activity, potentially is a metalloproteinase/disintegrin region that constitutes the disintegrin-like domains. Our findings demonstrate that the disintegrin domain of EC1.5 (a) lacks platelet aggregation inhibitory activity. On the contrary, this factor shows the property of a platelet aggregation inducer. Also, the EC5.1 (b) was observed as a single-band protein with a molecular mass of 7.5 kDa. EC5.1 (b) showed both anti-coagulant and anti-platelet properties. Additionally, the structure of the EC5.1 (b) fraction is expected to be similar to that of phospholipase A2, while EC4 (a) structure is potentially very similar to that of Echistatin with 5 kDa molecular mass. We introduce the predicted structure of P-II snake venom metalloproteinase/ disintegrin domains, phospholipase A2 and Echistatin-like fractions. Further research is therefore needed to determine the complete structure of these novel fractions and elucidate their mechanism of action and future therapeutic applications of cardiovascular and homeostasis disorders.

Viruses are obligate parasites and are completely dependent on host cells for survival and replication. RBK cell line developed and introduced by Razi Vaccine and Serum Research Institute, has been successfully established as a continuous cell line over successive passages.Viruses are obligate parasites that completely rely on host cells for survival and replication. Razi Bovine Kidney (RBK) cell line was introduced and developed by the Razi Vaccine and Serum Research Institute. It has been successfully established as a continuous cell line over successive passages. As demonstrated in this experimental study, the RBK cells have shown suitable sensitivity to certain viruses, including Bovine Herpesvirus-1 (BoHV-1) virus. In the present research, the RBK cell line characteristics were analyzed using molecular and karyotype methods and growth specifications. Cloning of the RBK cell line was performed using the limited dilution method, and each cell clone was quantitatively and qualitatively characterized. Four cell clones were compared based on their sensitivity to the BoHV-1 virus. Then, the RBK-D5 clone was selected as the most appropriate cell line for further studies. The RBK-D5 was subjected to tests for identity, chromosomal analysis, and doubling time. In the end, the origin of this cell line was confirmed by the PCR method. It was observed that the cell line exhibited karyotype diversity due to aneuploidy, which can be responsible for the procreation of chromosomal instability. This diversity represents chromosomal changes in the continuous cell line that carries the characteristic of an immortalized cell line. The RBK-D5 was found to be more sensitive to the BOHV-1 virus. Surprisingly, its titer was evaluated at 108.5 CCID50/ml. The obtained results suggested that the RBK cell line is suitable for the BoHV-1 virus and can be useful for virus detection, propagation, and quality control or viral titration.

Hemiscorpius species are distributed in Africa and Asia. Seven species of this genus have been identified in Iran of which six species have been reported from Hormozgan province. Members of this genus are the most dangerous scorpions in Iran. Scorpions were collected by moving stones during the day and searching at night using portable UV lights from 2011 to 2022 from different areas in Hormozgan province. Three species were identified from Hormozgan province including Hemiscorpius acanthocercus, H.enischnocela and H. shahii which are endemic to Iran. These species have limited distribution and were reported only from the south of Iran. The number of trichobothria are 3, 10-12 and 15-17 in H. acanthocercus, H. enischnochela and H. shahii, respectively. The measured values showed that H. Shahii is larger than H. enischnochela and H. acanthocercus. Specimens of H. acanthocercus are brown to dark brown samples with dark metasomal segment V. There have been reports of death from biting this species. Members of H. enischnochela are light brown to yellow samples. Members of H. Shahii are large brown samples. All three species have sexual dimorphism. Although these three species can be distinguished based on their morphological characters, the molecular investigation is needed to confirm the validity of all species of this genus. Identifying species and determining their distribution range is very useful in facilitating education and treatment management.

Giardia duodenalis, is one of the main causes of gastrointestinal disorders worldwide, which infects the small intestine of humans and animals. Based on the genetic characteristics of the parasite, eight genotypes (A to H) have been identified in clinical samples. The main purpose of the present study was to find the genetic diversity of Giardia in people referred to health centers in Semnan city using PCR. Totally, 300 stool samples were collected from people who were referred to health centers in Semnan city. The stool samples were first examined using the microscopic method (direct method and Lugol staining) and the samples were checked with trichrome staining. After DNA extraction, the GDH gene of positive samples was amplified by semi-nested PCR method. The genotype of positive samples was determined by the sequencing method. Out of 300 samples, only 20 (6.66%) were positive in the microscopic examination of the stool. In the PCR test, only 13 (4.33%) of the samples were positive. According to the multiple alignment results, it was found that the isolates belong to AII, BIII, and BIV genotypes. Most of which are related to people without clinical symptoms of diarrhea. Identification of AII, BIV, and BIII genotypes indicates the anthroponotic and anthropozoonotic transmission cycle of Giardia infection in Semnan city.

Foot-and-mouth disease is an extremely infectious and occasionally fatal viral disease with a rapid onset and a short course that affects cloven-hoofed animals and results in considerable financial losses. Today, Foot-and-mouth disease is controlled by traditional inactivated vaccines. Due to the short duration of immunity, a study was conducted for proteins of the virus as well as obtaining immunodominant proteins to design more efficient vaccines against Foot-and-mouth disease virus. This research aims to study the profile of Foot-and-mouth disease virus protein by electrophoresis and identification of the immunodominant proteins. The purified Foot-and-mouth disease virus was purchased then the protein concentration of that solution was measured by Lowry method. SDS-PAGE was done to achieve the protein profiles of the virus and immunization of 5 guinea pigs was done, then blood samples were taken for obtaining serum. Finally, serology tests; double immunodiffusion, ELISA, and western blotting were used to evaluate antigen response to antibodies (antigenic immunization). The protein concentration was 3.5 mg/ml. In SDS-PAGE with 10% gel, the protein profile of the virus was observed. After immunization, by conducting double immunodiffusion tests, the sediment lines between the serum antibody and the antigen of the virus were formed. Also, The ELISA test showed that the antibodies were formed against the antigens. In the western blot test, two immunodominant proteins of the FMD virus were obtained. According to the results, the immunodominant proteins of the FMD virus were determined. These proteins can be used in immunological diagnostic methods and also novel vaccines.

In this study, the Israeli acute paralysis virus (IAPV), a single-stranded RNA virus, was investigated in honey bee colonies, which had a history of mortality, population decline, and parasitic diseases. Samples (adult honey bees) were collected from 328 apiaries from three provinces (Tehran, Alborz, and Mazandaran) of Iran to detect IAPV. After sample preparation, RNA was extracted and cDNA was synthesized to perform the reverse transcription polymerase chain reaction (RT-PCR) method using a PCR primer pair, and a 185 bp fragment was amplified. The results showed that out of 328 samples, 103 (31.4%) samples were positive, which were from Mazandaran (14.33%), Tehran (8.84%), and Alborz (8.23%) provinces. Subsequently, some of the positive samples were sequenced and a phylogenetic tree was drawn. The phylogenetic tree showed that the virus isolates were divided into two distinct groups, including one group that had a high similarity to the European acute bee paralysis virus (ABPV) and one group that had a high similarity to the Kashmir bee virus. In addition, the sequences of the samples in three regions were separated in a node from the strains of ABPV from Eastern Europe. Since the length of the branch between the Iranian sequences and the different strains of ABPV from Eastern Europe was short, it can be assumed that the sequences from Iran have a common ancestor with the mentioned strains of ABPV from Eastern Europe.

Many researchers have been curious about the chemical sterilization method, which may be a choice of castration. The 20% calcium chloride ethanolic solution can prevent animals from some tumors and control the side effects of surgical castration. This experiment divided 12 male mixed-breed dogs into sham and chemical groups (n=6). Normal saline and 20% calcium chloride (20 ml/testis) were injected in the sham and chemical group's testis, respectively. Ultrasonography and related scoring were operated at 0-, 7-, 14-, and 2-days post-injection to evaluate echogenicity and measure the left testes' dimensions. Blood samples were taken on days 0, 7, 14, and 21 of the experiment evaluating the superoxide dismutase (SOD), glutathione peroxidase (GPx), and testosterone levels. The semen in the left epididymis of the chemical group was aspirated on day 21 post-injection for counting the sperm numbers. The testes of all dogs were surgically removed at 21 days post-injection, and the left one was put in formaldehyde for tissue processing. The intertubular edema, necrosis of the seminiferous tubules, neutrophil infiltration, and calcification was scored. The average dimensions of the chemical groups' left testes significantly decreased 7, 14, and 21 days after injection. The echogenicity of the testes decreased in the chemical group. A significant echogenicity difference was observed between the first day and the 7th and 14th day in ultrasonography. Calcium chloride injection failed to reduce the mean testosterone levels on all experimental days compared to day zero. Otherwise, the sperm number in the left testes of the chemical group decreased on day 21 post-injection. The degree of intertubular edema with neutrophil infiltration and severe tubular necrosis in the chemical group was significantly higher than in the sham group on the experimental days, including 7, 14, and 21. The mild calcification in the chemical group is likely the reason for higher echogenicity on day 21. The scrotum was swelled and ulcerated in the chemical group. Ultrasound is effective in demonstrating the castration ability of calcium chloride in the chemical method. Due to the inflammatory clinical effects, the chemical method is recommended in dogs only when surgical methods are unavailable.

The present research aimed to evaluate the addition of zinc (Zn) on antioxidant activity, blood profile, mineral availability, and abdominal fat of Sikumbang Janti duck. A total of 96 female Sikumbang Janti ducks aged 8 weeks were used in this research. This study used a completely random design with four treatments and four replications (6 duck/replications). The treatments were as follows: control diet (Z0), the addition of 30 mg Zn/kg (Z1), 60 mg Zn/kg (Z2), and 90 mg Zn/kg(Z3). Variables observed were antioxidant activity, blood profile, mineral content in the tibia, and abdominal fat. The results showed that Zn addition on feed significantly increased antioxidant activity (DPPH), Zn concentration in thigh, leukocytes, mineral availability (Ca, P, and Zn) (P<0.01), and decreased weight of abdominal fat in Sikumbang Janti duck (P<0.01). Blood profiles (except leukocytes) were not affected by the addition of Zn in the diet (P>0.05). It is concluded that the Z2 (60 mg Zn/kg) addition improves antioxidant activity, blood leukocytes, zinc content in thigh meat, mineral availability, and decreases abdominal fat weight of Sikumbang Janti duck.

Chronic heat stress affects numerous physiological and behavioral mechanisms. Epigenetic changes following prolonged cyclic heat stress, creating new opportunities for molecular biology research. One of these changes involves monoamines, such as serotonin, epinephrine, norepinephrine, dopamine, and their transmission. Broiler chickens are highly susceptible to heat stress, and their hearts become insufficient during the growth phase, leading to hypertrophy of the left heart. RNA-seq data were obtained from NCBI with accession number SRP082125. The expression level of genes was determined with DESeq2 packages. Gene Ontology qualification, including biological processes, cellular components, and molecular role (MF), was performed from the Gene Ontology Resource. Cyclic heat stress in broilers significantly altered monoamine receptor expression.  Twenty-nine genes of the monoamine pathway changed their expression in the left heart. Significant downregulation of expression was statistically associated with the ADRB1, HTR2A, and PNMT genes and upregulation of the MAOA gene (P<0.01). STRING database was used to construct the protein-protein interaction network; based on network analysis, the HTR2C, HTR2A, and HTR5A genes were identified as the major nodal genes in the network followed by MAOA, DRD2, DRD5, HTR1B, DRD1, DRD3, and HTR2B genes occupying the second important place in the network module. In conclusion, heat stress treatment prevented cardiac hypertrophy and altered the expression of monoamine genes. This would imply that monoamine transmission plays an important role in the development of cardiac hypertrophy, and that cyclic-chronic heat treatment modulates the cardiac monoaminergic system. These molecular biomarkers could be useful for screening, diagnosis, and treatment of cardiac hypertrophy.

Since depression is a common mental illness affecting an estimated 5% of people worldwide, investigators are encouraged to develop effective antidepressants. According to the monoamine-deficiency hypothesis, the underlying pathophysiology of depression is a deficiency of some neurotransmitters (serotonin, norepinephrine, or dopamine) in the central nervous system. The neurotransmitter serotonin has drawn the most attention concerning depression. As per research, 5-methoxy-N, N-dimethyltryptamine (5-MeO-DMT) elevates inter-synaptic serotonin levels when administered as a single inhalation of vapor from dried toad secretion and leads to higher life satisfaction, convergent thinking, higher ratings of mindfulness, lower ratings of depression, and anxiety. Furthermore, although 5-MeO-DMT lowers stress biomarkers such as cortisol, it is a psychedelic with hallucinogenic effects. In the present study, analogues of 5-MeO-DMT are designed with the hope that they might have better therapeutic activity and lower psychedelic side effects. The current study aimed to look at 5-MeO-DMT analogues as possible antidepressants. We used 70,000 5-MeO-DMT analogues that were sketched using Marvin to conduct a High Throughput Virtual Screening method in hopes of finding potential 5-MeO-DMT analogues against the 5-Hydroxytryptamine 1A receptor (5-HT1AR; 7E2Y.pdb) as an agonist. The prediction of the analogue-protein interaction and the evaluation of the binding affinity is accomplished by employing molecular docking. The Glide XP docking data indicated that a total of 21 compounds had Glide gscores ranging from -11.41 to -6.53 kcal/mol. When compared to the standard 5-MeO-DMT with the binding affinity of -7.75 kcal/mol, 14 compounds showed better binding affinity. Furthermore, Molecular Mechanics -Generalised Born and Surface Area solvation (MM-GBSA) indicated a binding free energy range of -63.55 to -35.37 kcal/mol, and 18 compounds showed better binding free energy than standard 5-MeO-DMT (-41.42 kcal/mol). Through ligand binding interactions with Asp116, Phe361, Phe362, Ser190, Ser199, Val117, Trp358, Ala365, Pro369, Ile189, Tyr195, Ala203, Ile167, Tyr390, Cys120, Trp358, Val364, Ala365, and Leu368, these complexes were stabilized, according to the molecular dynamic simulation of 20453/7E2Y in 100ns.

Salmonella is a zoonotic bacterium that is considered to be one of the most common causes of foodborne infections worldwide. Bearing in mind the genes involved in its virulence, identifying these genes can enable experts to better understand bacterial pathogenicity, which could subsequently help develop more efficient means to control and prevent infections. This study aimed to analyze stn, sipB, and sopB genes in various Salmonella serovars. To carry out this study, 103 Salmonella serovars were extracted from livestock, poultry, and humans from existing samples at the Department of Microbiology of the Razi Serum and Vaccine Research Institute in Karaj, Iran. These samples were cultured in selection and differential media, and their serovars were identified using specific antibodies based on Kaufman-White Tables. Utilizing PCR and specific primers, stn, sopB, and sipB genes were detected among these serovars. In this investigation, the most common human serovars were Salmonella paratyphi A, Salmonella paratyphi B, and Salmonella enteritidis; the most common serovars among livestock consisted of Salmonella dublin and Salmonella typhimurium and the most common Salmonella serovars among poultry consisted of Salmonella infantis and Salmonella enteritidis. The results of PCR on stn, sipB, and sopB genes demonstrated segments with 617bp, 875 bp, and 220 bp on agar gel, respectively. Based on the obtained findings, stn, sipB, and sopB genes were detected in 96.11%, 99.02%, and 98.05% of Salmonella serovars, respectively. Considering the fact that the aforementioned genes play significant roles in bacterial virulence, they can be used to develop diagnostic ELISA kits and recombinant vaccines.

Nanomaterials are characterized by mechanical, thermal, chemical, biological, and other properties that are different from the basic materials that make them up due to their large surface area to size ratio and quantum effect. There are multiple ways to produce nanomaterials mechanically, chemically, and physically, but they are not safe for the environment. Researchers have sought to find safe methods for the production of nanomaterials, such as green manufacturing, that is, manufacturing nanomaterials from plants. Moreover, there are other sources, such as bacteria or fungi that are used in the production of nanomaterials. This study aimed to try to find an alternative to chemically manufactured drugs, such as those used in the treatment of human cancers, through nanotechnology and from plant sources (green-biosynthesis), which is characterized by abundance and low economic cost. Silver nanoparticles were green-synthesized using an aqueous extract of the licorice plant, their properties were diagnosed, and their differences with the crude aqueous extract were determined. The sizes of nanoparticles were within the range of 60.27-89.80 nm, while the sizes of the crude aqueous extract particles were within the range of 53.96-113.1 nm. Atomic force microscopy was used to find out the shapes, topography, roughness, and protrusions of the surfaces of biosynthesized AgNPs and aqueous extract particles, where the roughness rate of the nanoparticles was 75.54 nm, while it appeared. In vitro test of AgNPs showed a higher anti-lung cancer activity against the A549 cell line than that of the extract at an inhibitory concentration for half of the cells used in the experiment (IC50) of 58.78 µg/ml while the IC50 of the extract was 67.44 µg/ml. The results showed that the toxicity of AgNPs on the normal hepatocyte line (WRL68) was less than that of the aqueous extract, with IC50 concentrations of 244.2 and 147.0 µg/ml, respectively. It is worth mentioning that the lower IC50 led to higher toxicity.

Antibiotic resistance is rising dramatically worldwide, and thus the production of new antibiotics is indispensable. Recent scientific initiatives have focused on the bioprospecting of microorganisms' secondary metabolites, with a particular focus on the look for natural products with antimicrobial properties derived from endophytes. All plant species, regardless of their type, are thought to anchor endophytic bacteria (EB). There are many potential uses for the natural therapeutic compounds made by EB in medicine, agriculture, and the pharmaceutical industry. To investigate antibacterial properties in this study, Actinomycetota (formerly, Actinobacteria) were isolated from Anthemis pseudocotula Boiss., identified, and underwent bioprospecting by morphological and molecular methods. Samples were collected from Ilam, Iran, and then divided into roots, leaves, stems, and flowers. After disinfection, they were cut into 2 mm pieces, cultured on casein agar culture medium, and incubated at 28°C for up to four weeks. Actinomycetota was identified using the polymerase chain reaction method targeting the 16S rRNA gene. To evaluate the antibacterial properties of the isolated Actinomycetota, the agar diffusion method was used. In parallel, the frequencies of biosynthetic gene clusters, including polyketide synthase (PKS-I and PKS-II) and nonribosomal peptide synthetase (NRPS) genes, were determined in the isolated Actinomycetota. Ninety bacteria were isolated from different parts of Anthemis flowers. Thirty-eight (42.2%) of these bacteria belonged to the phylum Actinomycetota, and out of these 38, 15 isolates (39.5%) had antibacterial properties. Of these, 11 isolates (73.3%) exhibited antibacterial effects against Staphylococcus aureus, 2 (13.3%) against Pseudomonas aeruginosa, 3 (20%) against Escherichia coli, and two isolates (13.3%) against Salmonella enterica sub-species of enterica serovar Typhimurium. The results of the molecular analysis of PKS-I, PKS-II, and NRPS genes showed that out of 38 isolated Actinomycetota strains, 23 isolates (60.5%) carried PKS-I gene, 6 (15.8%) harbored PKS-II gene, and 20 isolates (52.6%) had NRPS gene. This study indicates that Anthemis pseudocotula Boiss. has a number of active Actinomycetota that produce secondary metabolites with antibacterial properties.

The present study aimed to investigate the effect of supplementing broiler chickens’ diet with graded levels of moringa powder on growth performance. A total of 192 one-day-old broiler chicks were individually weighed and randomly distributed into four dietary treatments. Each treatment comprised four replicates with 12 chicks in each. Moringa powder was added to their diet by 0.0%, 0.25%, 0.5%, and 0.75%. The diet and water were offered ad libitum during the feeding trial, which lasted 42 days. One chicken was selected from each replicate at the end of the experiment to measure the carcass characteristics and meat quality, as well as the serum biochemical parameters of broilers. Regarding the overall growth performance, body weight gain and feed conversion ratio substantially improved (P<0.05) in broilers whose diet was supplemented with moringa powder, compared to the control group. Furthermore, the carcass yield considerably increased (P<0.05) in broilers whose diet was supplemented with 0.5% and 0.75% moringa powder, in comparison with the control group. In addition, birds fed with a diet supplemented with moringa powder showed a significant increase in their hemoglobin level (P<0.05). Moreover, the findings showed that a diet supplemented with moringa powder led to a significant decrease in the total cholesterol level, low-density lipoprotein, and the A/G ratio (P<0.05) but increased total protein and globulin levels (P<0.05), compared to the control group. In conclusion, the supplementation of 0.75% moringa powder in the diet as a growth promoter reduces the cost of production by improving growth performance and enhancing the health status of birds.

This study aimed to evaluate the effectiveness of direct and indirect modified ISO 10272-1:2017 methods for detecting Campylobacter spp. in 10 sites of a poultry slaughterhouse and investigate the relationship between poultry intestinal carriage and carcasses, as well as surfaces contamination during different slaughter steps (scalding, defeathering, evisceration, and rinsing). Antibiotic resistance profiles of the isolates were also determined against 12 antibiotics. A total of 165 intestinal (feces and ceca) and non-intestinal (neck skins and surfaces) samples were collected from 10 different sampling sites before, during, and after the slaughtering of six flocks of broiler chickens. After the isolation and phenotypic identification of the isolates, an antibiotic susceptibility study was performed using the agar diffusion method. Thermotolerant bacteria of the genus Campylobacter (TC) were isolated with a prevalence of 47.04% (127/270), and 39.05% (82/210) of the TC isolates were detected in non-intestinal samples. Moreover, 76.19% (80/105) of these microorganisms were detected by a direct isolation method for a sensitivity of 97.56%, while only 1.90% (2/105) of the samples contained TC by an indirect isolation method for a sensitivity of 2.44%. The samples of intestinal origin were positive for TC with a rate of 75.00% (45/60). C. jejuni (76.38%; 97/127) was the most isolated bacterial species. Furthermore, 98.43% (125/127) of the TC isolates were multidrug-resistant and 69.29% (88/127) showed simultaneous resistance to ciprofloxacin and erythromycin. Direct isolation seems to be the best method for the detection of C. spp. A serious public health problem of multidrug-resistant C. spp. isolates with critical resistance profiles can be transmitted to broiler carcasses before, during, and after the evisceration step.