Development of a novel Colorimetric Assay for the rapid diagnosis of Coronavirus disease 2019 from nasopharyngeal samples.

Emergence of Coronavirus disease 2019 (COVID-19) pandemic has posed a huge threat to public health. Rapid and reliable test to diagnose infected subjects is crucial for disease spread control. We developed a colorimetric test for COVID-19 detection using a Colorimetric Assay based on thiol-linked RNA modified gold nanoparticles (AuNPs) and oligonucleotide probes. This method was conducted on RNA from 200 pharyngeal swab samples initially tested by Real-Time polymerase chain reaction (RT-PCR) as gold standard. A specific oligonucleotide probe designed based on ORF1ab of COVID-19 was functionalized with AuNPs-probe conjugate. The exposure of AuNP-probe to isolated RNA samples was tested using hybridization. In this comparative study, the colorimetric functionalized AuNPs assay exhibited a detection limit of 25 copies/µL. It was higher in comparison to the RT-PCR method, which could only detect 15 copies/µL. The results demonstrated 100% specificity and 96% sensitivity for the developed method. Herein, we developed an incredibly rapid, simple and cost-effective Colorimetric Assay lasting approximately 30 min which could process considerably higher number of COVID-19 samples compared to the RT-PCR. This AuNP-probe conjugate colorimetric method could be considered the optimum alternatives for conventional diagnostic tools especially in over-populated and/or low-income countries.

Evaluation of the Expression of Infection-Related Long Noncoding RNAs among COVID-19 Patients: A Case-Control Study.

Background and Aims: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a worldwide pandemic, activates signaling cascades and leads to innate immune responses and secretion of multiple chemokines and cytokines. Long noncoding RNAs (lncRNAs) have a crucial role in inflammatory pathways. Through our search on the PubMed database, we discovered that existing research has primarily focused on examining the regulatory impacts of five lncRNAs in the context of viral infections. However, their role in regulating other conditions, including SARS-CoV-2, has not been explored. Therefore, this study aimed to investigate the expression pattern of lncRNAs in the peripheral blood mononuclear cells (PBMC) and their potential roles in SARS-CoV-2 infection. Potentially significant competing endogenous RNA (ceRNA) networks of these five lncRNAs were found using online in-silico techniques. Methods: Ethylenediaminetetraacetic acid (EDTA) blood samples of the control group consisted of 45 healthy people, and a total of 53 COVID-19-infected patients in case group, with a written informed consent, was collected. PBMCs were extracted, and then, the RNA extraction and complementary DNA (cDNA) synthesis was performed. The expression of five lncRNAs (lnc ISR, lnc ATV, lnc PAAN, lnc SG20, and lnc HEAL) was assessed by real-time PCR. In order to evaluate the biomarker roles of genes, receiver operating characteristic (ROC) curve was drawn. Results: Twenty-four (53.3%) and 29 (54.7%) of healthy and COVID-19-infected participants were male, respectively. The most prevalent symptoms were as follows: cough, general weakness, contusion, headache, and sore throat. The results showed that three lncRNAs, including lnc ISR, lnc ATV, and lnc HEAL, were expressed dramatically higher in the case group compared to healthy controls. According to ROC curve analysis, lnc ATV has a higher AUC and is a better biomarker to differentiate COVID-19 patients from the healthy controls. Then, using bioinformatics methods, the ceRNA network of these lncRNAs enabled the identification of mRNAs and miRNAs with crucial functions in COVID-19. Conclusion: The considerable higher expression of ISR, ATV, and HEAL lncRNAs and the significant area under curve (AUC) in ROC curve demonstrate that these RNAs probably have a potential role in controlling the host innate immune responses and regulate the viral replication of SARS-CoV-2. However, these assumptions need further in vitro and in vivo investigations to be confirmed.

A study of rural populations' knowledge, attitude, and practice about brucellosis: a descriptive, cross-sectional, multicenter study.

OBJECTIVE: Brucellosis is a highly contagious disease which is transmitted from animals to humans. One of the populations at high risk of infection is those living in rural areas. The present study was conducted to investigate rural populations' knowledge, attitude, and practice about brucellosis in Iran. The study used a descriptive, cross-sectional design to assess 300 individuals who were living in rural areas. The subjects were selected using convenience sampling from six villages located in the south of Iran. The data gathered were analyzed using Analysis of variance (ANOVA), and Pearson correlation coefficient in SPSS version 23. RESULTS: From the 300 individuals who were enrolled in this study, 189 were male and 111 were female. The mean age of the participants was 48.27 ± 4.28 years. The mean scores of the study population's knowledge, attitude, and practice about brucellosis were found to be low. A significant direct correlation was found between the subjects' knowledge, attitude, and practice regarding brucellosis.

Promising Larvicidal Effects of Nanoliposomes Containing Carvone and Mentha spicata and Tanacetum balsamita Essential Oils Against Anopheles stephensi.

PURPOSE: The use of synthetic pesticides to control the spread of mosquito-borne diseases has caused environmental pollution and insecticide resistance in mosquitoes. Developments of new green insecticides have thus received more attention to overcome these problems. METHODS: Nanoliposomes containing carvone and essential oils were first prepared. The nanoliposome physicochemical characteristics (particle size, morphology, and successful loading) were then evaluated by Dynamic Light Scattering (DLS), Transmission Electron Microscopy (TEM), and the Attenuated Total Reflection-Fourier Transform InfraRed (ATR-FTIR) analyses. Larvicidal effects of carvone, Mentha spicata, and Tanacetum balsamita essential oils were investigated against the main malaria vector, Anopheles stephensi, in non-formulated and nanoformulated states. RESULTS: The larvicidal effects of nanoformulated states were significantly more potent (7.2 folds, 3.5 folds, and 8 folds) than non-formulated states. Nanoliposomes containing M. spicata and T. balsamita essential oils with particle sizes of 175 ± 8 and 184 ± 5 nm showed the best efficacies (LC50 values = 9.74 and 9.36 µg/mL). CONCLUSION: The prepared samples could be used as new green potent larvicides against An stephensi mosquito in further field trials. It is also recommended to investigate their efficacies against other mosquitoes.

Encapsulation of probiotic Lactobacillus acidophilus ATCC 4356 in alginate-galbanum (Ferula Gummosa Boiss) gum microspheres and evaluation of the survival in simulated gastrointestinal conditions in probiotic Tahini halva.

One of the famous traditional confectionery products is Tahini halva. The aim of this study was the production of probiotic halva using free Lactobacillus acidophilus (FLA) and microencapsulated Lactobacillus acidophilus (MLA) with sodium alginate and galbanum gum as the second layer. The survival rate of MLA and FLA during heat stress, storage time, and simulation gastrointestinal condition in Tahini halva was assessed. The survival rates of MLA and FLA under heat stress were 50.13% and 34.6% respectively. During storage in Tahini halva, the cell viability loss was 3.25 Log CFU g-1 and 6.94 Log CFU g-1 for MLA and FLA, separately. Around 3.58 and 4.77 Log CFU g-1 bacteria were reduced after 6 h of exposure in simulated gastrointestinal conditions, for MLA and FLA respectively. These results suggest that the use of alginate and galbanum gum is a promising approach to protecting L. acidophilus against harsh environmental conditions.

Muscular hydatid cyst in Iran: A case report.

INTRODUCTION AND IMPORTANCE: Hydatid disease, caused by the larval stage of Echinococcus granulosus, is a common parasitic infection of humans and herbivores. Although livers and lungs are the most commonly affected organ, hydatid cysts may develop in any body part. Primary muscular hydatid cyst is extremely rare. CASE PRESENTATION: We reported the case of a 40-year-old-woman with the presentation of a soft, mobile, and non-tender lump in the dorsal part of her left upper arm (triceps brachii), which emerged one year ago. Her past medical history was unremarkable. The arm sonography revealed a single uniloculated cystic mass (6.5 cm × 5.5 cm) with a thick wall containing cystic lesions. It suggested the diagnosis of echinococcosis. The patient underwent surgery, and the hydatid cyst was excised. Histopathological examination confirmed hydatidosis. CLINICAL DISCUSSION: Hydatid cysts occur rarely (about 4 %) in muscles even in endemic regions. The study is the first case of hydatidosis found in triceps brachii in Fars province, Iran. In endemic regions, considering the hydatid cyst possibility is very important because it presents with many diversities. As it clinically presents a painless slow-growing mass, may be misdiagnosed with benign soft tissue tumors. CONCLUSION: Although muscular hydatidosis is extremely rare, it should be considered a differential diagnosis of any growing subcutaneous or muscular masses or tumors. Imaging modalities and blood tests are highly relevant for diagnosis. Surgical excision, a choice of treatment, should be done with cautions and is combined with anthelmintic therapy to reduce the risk of recurrence.

A Nanoliposomal Gel Containing Cinnamomum zeylanicum Essential Oil with Effective Repellent against the Main Malaria Vector Anopheles stephensi.

Malaria is the most important vector-borne disease; however, mosquito repellents are still a practical approach for controlling malaria, especially in endemic regions. Due to the side effects of synthetic repellents such as N, N-diethyl-meta-toluamide (DEET), the development of natural repellents has received much attention. In this study, nanoliposomes containing 0.5 and 2.5% w/v Cinnamomum zeylanicum essential oil were firstly prepared with particle sizes of 119 ± 6 and 195 ± 9 nm. Their morphologies and loading of the essential oil in the particles were then investigated using transmission electron microscopy (TEM) and attenuated total reflection-Fourier transform infrared (ATR-FTIR) analyses. The nanoliposomes were finally jellified to increase their viscosity and facilitate topical usage. The complete protection time of the nanoliposomal gel containing 2.5% C. zeylanicum essential oil was significantly longer than that of 2.5% DEET against Anopheles stephensi: 303 ± 10 > 242 ± 12 min, p < 0.001. Moreover, the prepared nanoformulation was stable for at least six months at 4 and 26°C. Therefore, the prepared prototype could be considered a natural repellent against the main malaria mosquito vector in field conditions. In addition, it is suggested to be investigated against other important factors mosquitoes.

Individual genetic variability mainly of Proinflammatory cytokines, cytokine receptors, and toll-like receptors dictates pathophysiology of COVID-19 disease.

Innate and acquired immunity responses are crucial for viral infection elimination. However, genetic variations in coding genes may exacerbate the inflammation or initiate devastating cytokine storms which poses severe respiratory conditions in coronavirus disease-19 (COVID-19). Host genetic variations in particular those related to the immune responses determine the patients' susceptibility and COVID-19 severity and pathophysiology. Gene polymorphisms such as single nucleotide polymorphisms (SNPs) of interferons, TNF, IL1, IL4, IL6, IL7, IL10, and IL17 predispose patients to the severe form of COVID-19 or severe acute respiratory syndrome coronavirus-2 (SARS-COV-2). These variations mainly alter the gene expression and cause a severe response by B cells, T cells, monocytes, neutrophils, and natural killer cells participating in a cytokine storm. Moreover, cytokines and chemokines SNPs are associated with the severity of COVID-19 and clinical outcomes depending on the corresponding effect. Additionally, genetic variations in genes encoding toll-like receptors (TLRs) mainly TLR3, TLR7, and TLR9 have been related to the COVID-19 severe respiratory symptoms. The specific relation of these mutations with the novel variants of concern (VOCs) infection remains to be elucidated. Genetic variations mainly within genes encoding proinflammatory cytokines, cytokine receptors, and TLRs predispose patients to COVID-19 disease severity. Understanding host immune gene variations associated with the SARS-COV-2 infection opens insights to control the pathophysiology of emerging viral infections.

Isolation and characterization of E. coli O157: H7 novel bacteriophage for controlling this food-borne pathogen.

E. coli O157: H7 is known as a high-risk food-born pathogen, and its removal is vital for maintaining food safety. The increasing trend of food-borne diseases caused by this bacterium and other pathogens indicates the low efficiency of the methods to remove pathogens from foodstuffs. One of the new and effective methods is to use of a bio-control agent called bacteriophage, which has shown good function in eliminating and reducing pathogens. In this study, a novel bacteriophage was isolated and identified from the slaughterhouse wastewater to control E. coli O157: H7. This bacteriophage belonged to the Myoviridae family. Two bacterial genera including E. coli and Salmonella, were allocated to determine the bacteriophage host range; the result showed that the anti- Salmonella effect of phage was low. The phage was stable at high temperature (80 °C) and caused an acceptable reduction in the E. coli O157: H7 (4.18 log CFU / mL for 10 h). The isolated bacteriophage was corroborated to be completely safe based on the whole genome sequencing and lack of any virulence factor from the host bacteria. Considering the characteristics of this phage and its function in vitro, this bacteriophage may be used as an effective bio-control agent in foods with the possible E. coli O157: H7 -induced contamination.