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Low-density lipoprotein receptor-related protein 6 (LRP6) is an important therapeutic target for diseases such as osteoporosis, Alzheimer, cancer, and neurodegenerative disease. Computational methods such as ligand-based and structure-based virtual screening have been introduced as an extremely efficient and accurate tool for finding new drug targets and candidates. In this study, we aimed to screen the National Cancer Institute (NCI) Diversity Set II and parts of the ZINC database by virtual screening to identify potential and safe compounds that can inhibit the LRP6 protein. By utilizing various screening methods such as rigid and flexible molecular docking and Lipinski's rule of five, we identified 10 potential compounds. Then, their validity was further tested by molecular dynamics simulation and MMPBSA binding free energy calculations. Eventually, it was concluded that ZINC03954520, ZINC01729523, ZINC03898665, ZINC13152226, ZINC26730911 and ZINC01069082 compounds can be potential drug compounds for inhibiting LRP6 protein. These compounds in complex with β-propeller domains of LRP6 showed that they are capable of altering the backbone of these domains and interfere with their structural dynamics which may lead to the inhibition of the signal transmission. Communicated by Ramaswamy H. Sarma.

Introduction. Herpes simplex virus type 1 (HSV-1) is a virus that causes serious human disease and establishes a long-term latent infection. The latent form of this virus has shown to be resistant to antiviral drugs. Clustered Regularly Interspace Short Palindromic Repeats (CRISPR), is an important tool in genome engineering and composed of guide RNA (gRNA) and Cas9 nuclease that makes an RNA-protein complex to digest exclusive target sequences implementation of gRNA. Moreover, CRISPR-Cas9 system effectively suppresses HSV-1 infection by knockout of some viral genes. Materials and methods. To survey the efficacy of Cas9 system on HSV-1 genome destruction, we designed several guide RNAs (gRNAs) that all packaged in one vector. Additionally, we performed a one-step restriction using BamHI and Esp3I enzymes. Results. CRISPR/Cas9 system targeted against the gD gene of HSV-1 was transfected into HEK-AD cells that showed a significant reduction of HSV-1 infection by plaque assay and real-time PCR. Conclusion. The pCas-Guide-EF1a-GFP CRISPR vector can create a fast and efficient method for gRNA cloning by restriction enzymes (Esp3I (BsmBI) and BamHI). Therefore, the CRISPR/Cas9 system may be utilized for the screening of genes critical for the HSV-1 infection and developing new strategies for targeted therapy of viral infections caused by HSV-1.

Toxoplasma gondii is a common protozoan parasite, which infects warm-blooded mammals, including mice and humans, throughout the world. The negative effects of T. gondii infection on the human reproductive system have been documented, especially in females. However, only few studies have examined the effects of T. gondii infection on the male reproductive system. Previous research shows that T. gondii can induce DNA methylation in some gene promoters, which are key regulators of spermatogenesis. Therefore, this study aimed to evaluate the effects of curcumin on the activity of DNA methyltransferases (DNMTs), as well as selected genes, involved in spermatogenesis in spermatogenic cells. In the spermatogenic cells exposed to T. gondii, there was a significant increase in DNMT1 and DNMT3A gene expression and a significant reduction in HSPA1A, MTHR, and DAZL gene expression, compared to the controls. The present results showed that curcumin could regulate changes in T. gondii-mediated gene expression. The effect of T. gondii on DNMT activity was also investigated in this study. A 40 % increase in DNMT activity was observed due to T. gondii infection. However, DNMT activity was restored by treatment with 20 μM curcumin for eight hours. The results revealed that T. gondii increases the NF-κB activity, compared to the control group. The increase in NF-κB activity, induced by T. gondii, was inhibited by curcumin. In conclusion, T. gondii, by increasing DNMT expression and activity, leads to an increase in NF-κB activity in cells. On the other hand, curcumin reduced DNA methylation, induced by T. gondii, owing to its NF-κB-inhibiting properties. Therefore, curcumin, as a hypomethylating agent, can be potentially used to alleviate the negative effects of T. gondii on the male reproductive system., EXCLI Journal;Vol. 19. 2020, pp.

Background: miRNAs are small non-coding RNAs with potential roles in the complications of pregnancy. We hypothesised links between polymorphisms in miRNA-196a2 and miRNA-499 in maternal blood and t...

Objective(s): Oncolytic Herpes simplex virus type 1 (HSV-1) has emerged as a promising strategy for cancer therapy. However, development of novel oncolytic mutants has remained a major challenge owing to low efficiency of conventional genome editing methods. Recently, CRISPR-Cas9 has revolutionized genome editing.Materials and Methods: In this study, we aimed to evaluate the capability of CRISPR-Cas9 to manipulate the UL39 gene to create oncolytic HSV-1. Herein, three sgRNAs were designed against the UL39 gene and transfected into HEK-293 cell line followed by infection with HSV-1 KOS.Results: After three rounds of plaque purification, several HSV-1 mutants were identified by PCR analysis and sequencing. One of these mutations in which 55 nucleotides were deleted resulted in a frameshift mutation that in turn produced a truncated protein with only 167 amino acids from 1137 amino acids. Functional analysis in Vero and primary fibroblast cells revealed that viral replication was significantly lower and plaque size was smaller in the HSV-1 mutant compared with HSV-1 KOS. Moreover, the relative amount of viral genome present in the supernatants of infected cells (Vero and primary fibroblast cells) with HSV-1 mutant was significantly decreased compared with those of HSV-1 KOS.Conclusion: Our data revealed that targeting UL39 with CRISPR-Cas9 could develop oncolytic HSV-1.

Introduction: One of the goals of the health system is to ensure that people are healthy. Reaching this goal is only possible through the provision of the proper services. The satisfaction of patients with the quality of health care services is used as a questionare to investigate the effectiveness of hospital services concerning consumption management and patient care quality. It assists managers and the decision-making network to identify the strengths and weaknesses of the system. Objective: This study aimed to investigate the satisfaction of hospitalized patients in educational and medical centers of Rasht City, Iran with the “Health Care System Reform Plan” in 2019. Materials and Methods: The present study is an analytical cross-sectional study. A total of 395 patients hospitalized in educational and medical centers of Rasht City were selected based on the stratified random sampling method and according to the inclusion criteria. The study data were collected by a questionnaire examining patientschr('39') satisfaction with the “Health Care System Reform Plan” developed by the Treatment Deputy of Guilan University of Medical Sciences. The questionnaire has 37 questions in 5 areas of patient’s satisfaction with hoteling, hospital bills reduction program, resident physician program, notification program, the program for the promotion of normal childbirth. Data analysis performed using descriptive statistics indicators and Mann-Whitney U, Kruskal Wallis, and logistic regression model. Results: The results showed the mean±SD of sampleschr('39') age was 43.51±18 years. The majority (23.8%) of the samples were less than 30 years old. Moreover, there was a significant difference between satisfaction score of the Plan (P = 0.009) in terms of gender, hospital bills (P = 0.001), resident physicians (P = 0.009), and notification program (P = 0.032). Also, men had a higher overall score than women. The findings of this study showed that the mean ± SD satisfaction was equal to 82.8 9.1 with a minimum score of 39.71. In the logistics model, only two variables of the hospital (P = 0.017) and job (P = 0.001) were considered as the most important factors related to the overall satisfaction with the “Health Care System Reform Plan.” Conclusion: Based on the results of the study, patientschr('39') satisfaction with the implementation of the “Health Care System Reform Plan” was at the acceptable level and various factors have contributed to this level of satisfaction.

The volatile organic compounds (VOCs) have toxic effect on human health and environmental matrixes. So, determination and removal VOCs from the environmental samples such as water, wastewater and air are very important as toxicology effect on humans. Many chemistry techniques such as; analytical methods for sorbents (extraction, adsorption), sole gel method, pervaporation, regenerative catalytic oxidation (RCO), recuperative catalytic oxidation (CO), adsorptive concentration-catalytic oxidation, photocatalytic oxidation (PCO), ozonation-catalytic oxidation and non-thermal plasma-catalytic oxidation, were used for removal and decreasing of VOCs from different matrix. This review study introduces the adsorbents and applied chemistry methods which were recently used in different works for removal of VOCs in air or water samples by scientists.

Changes in the concentrations of trace metals such as zinc (Zn) and selenium (Se) can pathologically lead to neurodegenerative conditions such as the Alzheimer's disease (AD). Previous studies have shown that mitochondrial dysfunction plays an important role in the pathogenesis of AD. Several male Wistar rats were randomly divided into five groups: sham group, AD group that received 3 mg/kg of streptozotocin (STZ) intracerebroventricularly, AD + Zn group that received 10 mg/kg of Zn intraperitoneally (i.p.) for 1 week, AD + Se group that received 0.1 mg/kg of Se i.p. for 1 week, and AD + Zn + Se group that received 10 mg/kg of Zn i.p. plus 0.1 mg/kg of Se i.p. for 1 week. At end of the study, behavioral tests and mitochondrial oxidative stress and GPR39 gene expression evaluations were carried out. Co-administration of Zn and Se significantly decreased the potential collapse of mitochondrial membrane, reactive oxygen species levels, and lipid peroxidation levels while significantly increased cognitive performance, superoxide dismutase (SOD), glutathione peroxidase, and catalase activity in the brain mitochondria compared with the STZ group. In addition, no significant changes were observed in GPR39 expression in the co-treated group. Findings of the current study showed that ZnR/GPR39 receptor, mitochondrial dysfunction, and oxidative stress play important roles in the pathogenesis of AD. Co-treatment of Zn and Se improved the cognitive performance, mitochondrial dysfunction, and oxidative stress caused by STZ-induced AD. Therefore, therapeutic approaches to improve mitochondrial function could be effective in preventing the initiation and progression of AD.

3C protease is one of the most important proteases in enteroviruses, which is responsible for several cleavages of the viral polyprotein, leading to virus maturation. It has several harmful effects on host cells, such as the induction of caspase-mediated apoptosis and the cleavage of various proteins like P53, TATA-binding protein (TBP), histone H3, octamer-binding protein 1 (Oct-I) and DNA polymerase III. In this study, we aimed to construct a structure to express enterovirus 71 3C protease, consisting of a 3C protease sequence and two kinds of linkers including rigid and cleavable. As a result of cloning the structure into a pEGFP-N1 vector, two forms of constructions were expressed and verified by western blotting analysis including a 3C-Linker-Cleavage site (3C-L-CL) and a 3C-Linker (3C-L) without a cleavage site. In 3C-L-CL construction, 3C protease is released from fused GFP by cleave cleavage site. In 3C-L construction, 3C protease was expressed in fusion with GFP. In fact, one of the constructions produced two forms of 3C proteases, a free 3C and a 3C fused with GFP. This study shows that free 3C is preferred over the second form (3C with linker) because it is not connected to other peptides, and its protease activity is not affected. The designed structures could be used as useful tools for 3C protease expression and investigation of its interactions with cellular signaling pathways such as interferon pathway.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was first identified in several patients who traveled to Wuhan or went to a seafood wholesale market in Wuhan. The phylogenetic tree showed that SARS-CoV-2 was 96.2% identical to bat β-coronaviruses from lineage B. Also, several studies reported that SARS-CoV-2 uses the SARS-CoV receptor, angiotensin-converting enzyme 2, for entry to target cells. Lung alveolar and small intestine are potential targets for SARS-CoV-2 due to the high expression of the angiotensin-converting enzyme 2 receptor. In this review, we focused on the zoonotic β-coronaviruses and given there is no specific drug or vaccine for coronavirus disease 2019, we reviewed the literature on the therapy options for SARS and Middle East respiratory syndrome coronavirus infection, in order to discover their possible use in the treatment of SARS-CoV-2 infections.

Nabi Jomehzadeh,1– 3 Hazhir Javaherizadeh,4 Mansour Amin,2,3 Mohammad Rashno,1 Ali Teimoori5 1Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 2Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Department of Microbiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 4Alimentary Tract Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Khouzestan, Iran; 5Department of Virology, Faculty of Medicine, Hamedan University of Medical Sciences, Hamedan, IranCorrespondence: Mansour AminInfectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranTel +98 9163061096Fax +98 61-3333 2036Email mnsamin@yahoo.comBackground: Functional constipation is a common and annoying gastrointestinal disorder among children worldwide in which the intestinal microbiota composition plays a fundamental role. This study aimed to compare the quantity of main intestinal Lactobacillus species in constipated children and healthy controls.Materials and Methods: Fecal samples were collected from 40 children fulfilling Rome IV criteria for functional constipation and 40 healthy volunteers. Quantitative real-time polymerase chain reaction (qPCR) method with species-specific primers was used to investigate seven main Lactobacillus species in fecal samples.Results: Lactobacillus strains of the patient group were different from the healthy controls, main differences being noticeable decrease in the population quantity of Lactobacillus reuteri (mean 102.61 CFU/gram feces) and lower abundance of Lactobacillus fermentum (p< 0.0001).Conclusion: Lactobacillus species might be quantitatively altered in childhood functional constipation. Our findings on the different species of Lactobacillus population showed significantly decreased quantity in the patient group compared with the healthy subjects.Keywords: constipation, Lactobacillus, microbiota, quantitative real-time PCR

Background Functional constipation is a common and annoying gastrointestinal disorder among children worldwide in which the intestinal microbiota composition plays a fundamental role. This study aimed to compare the quantity of main intestinal Lactobacillus species in constipated children and healthy controls. Materials and Methods Fecal samples were collected from 40 children fulfilling Rome IV criteria for functional constipation and 40 healthy volunteers. Quantitative real-time polymerase chain reaction (qPCR) method with species-specific primers was used to investigate seven main Lactobacillus species in fecal samples. Results Lactobacillus strains of the patient group were different from the healthy controls, main differences being noticeable decrease in the population quantity of Lactobacillus reuteri (mean 102.61 CFU/gram feces) and lower abundance of Lactobacillus fermentum (p

Rotavirus is the most important etiological agent of infectious diarrhea in children under 5 years of age with more than 125,000 deaths occurring annually worldwide. The present study aims to determine the effect of curcumin, a natural polyphenol compound, on rotavirus in a cell culture model. The anti-viral activity of curcumin was evaluated by reverse-transcriptase quantitative PCR (RT-qPCR), TCID50, and western blot techniques to assess CC50 in curcumin-treated MA104 cells as well as EC50 and SI within the infected MA104 cell line. Our findings supported that curcumin exerted an inhibitory influence against rotavirus in a dose-dependent manner and decreased the viral titer and VP6 expression by ~99% at a concentration of 30 μM (p Keywords: curcumin; rotavirus; RT-qPCR; in vitro; anti-rotavirus agent.

In this paper, we first review the weighted-versiion of the handshaking lemma based on the idea of a weighted vertex-edge incidence matrix of a given graph $G$. Then, we obtain a generalized version of the handshaking lemma based on the concept of the clique value. We also define a generalized version of Randic index. More importantly, we prove an upper bound for the generalized Randic index of a graph $G$. We finally concluse the paper with some disscussions about possible future works., 7 pages

Background and Objectives: The human papillomavirus (HPV) is associated with more than 70% of the cervical neoplasm. The current study aims to evaluate the distribution of HPV genotypes in suspected women cytological specimens from Teh- ran, Iran. Materials and Methods: In the current cross-sectional study, HPV genotype prevalence was investigated in 433 subject women. DNA extraction was performed by High Pure Viral Nucleic Acid kit. A semi-automatically hybriSpot 24™ (HS24) setting was used for HPV typing and data interpreted by hybriSoft™ software according to instructions. Results: Pathologic data showed 181 (41.8%) had non-malignant lesions, 212 (49%) had inflammation and 40 (9.2%) reported LSIL in primary Pap-smear result. HPV was found in 143 (33%) specimens and the most comment high-risk and low-risk HPV types were HPV-16 and -6, respectively. Also, 62 (43%) were co-infected with multiple genotypes includes, 34 (24%) cases had co-infection with two HPV types, 17 (12%) cases had co-infection with three HPV types, 6 (4%) cases had co-infection with four HPV types and 5 (3%) cases had co-infection with five HPV types. There was statistically different domination on high-risk genotype in most of the co-infected samples (p

The ordinary generating function of the number of complete subgraphs of $G$ is called a clique polynomial of $G$ and is denoted by $C(G,x)$. A real root of $C(G,x)$ is called a clique root of the graph $G$. Hajiabolhasan and Mehrabadi showed that the clique polynomial has always a real root in the interval $[-1,0)$. Moreover, they showed that the class of triangle-free graphs has only clique roots. Here, we generalize their result by showing that the class of $K_4$-free chordal graphs has also only clique roots. Moreover, we show that this class has always a clique root $-1$. We finally conclude the paper with several important questions and conjectures., 7 pages

The authors have requested that this preprint be removed from Research Square.

In this short note, we first associate a new simple undirected graph with a given word over an ordered alphabet of $n$-letters. We will call it the Lyndon graph of that word. Then, we introduce the concept of the Lyndon-word representable graph as a graph isomorphic to a Lyndon graph of some word. Then, we introduce the generalized Stirling cycle number $S(N;n,k)$ as the number words of length $N$ with $k$ distinct Lydon words in their Lyndon factorization over an ordered alphabet of $n$-letters . Finally, we conclude the paper with several interesting open questions and conjectures for interested audiences., 5 pages

The ordinary generating function of the number of complete subgraphs (cliques) of $G$, denoted by $C(G,x)$, is called the The clique polynomial of the graph $G$. In this paper, we first introduce some \emph{clique} incidence matrices associated by a simple graph $G$ as a generalization of the classical vertex-edge incidence matrix of $G$. Then, using these clique incidence matrices, we obtain two clique-counting identities that can be used for deriving two combinatorial formulas for the first and the second derivatives of clique polynomials. Finally, we conclude the paper with several open questions and conjectures about possible extensions of our main results for higher derivatives of clique polynomials.

In this paper, we first give a simple combinatorial proof of Tepper's identity. Then, as a by product of this interesting identity we present another proof of the well-known Wilson's identity in number theory. Finally, we obtain a generalization of Tepper's identity for any polynomial with real coefficients., 9 pages, no figures

Background: Epigenetic alterations such as DNA methylation are known as the main cause of different types of cancers through inactivation of tumor suppressor genes, especially thyroid cancer. Identification of novel and effective markers are important in diagnosis and prevention of thyroid cancer. In the present study, the expression and methylation of Solute carrier family 5 member 8 (SLC5A8) in Papillary Thyroid Carcinoma (PTC) in comparison to multinodular goiter (MNG) have been studied. Methods: Overall, 41 patients with PTC and 36 patients affected by MNG were recruited from four hospitals in Tehran and Qazvin, Iran in 2018. Thyroid tissues were obtained during thyroidectomy. RNA and DNA were extracted from thyroid tissues. Quantitative RT-PCR assay was performed for determining the mRNA level of SLC5A8 while Methylation-Sensitive High-Resolution Methylation was applied for assessing the methylation status. Results: Methylation status of three regions composed of 52 CpG islands in the promoter of SLC5A8 gene was studied by HRM assay. SLC5A8 level in PTC tissues was significantly downregulated in average 0.4 fold in comparison with MNG tissues (P=0.05). The aberrant methylation of SLC5A8 (b) region was remarkably different in PTC and MNG cases. The promoter methylation of SLC5A8 (c) was significantly related to BRAF mutations and vascular invasion in PTC patients. Conclusion: The aberrant promoter hyper methylation of SLC5A8 was related to aggressive PTC. Therefore, there is some evidence to support the hypothesis that SLC5A8 could be a paly important role in the development of PTC.

Globally, it is estimated that 43 million people are living with human immunodeficiency virus type 1 (HIV-1), and there are more than 600,000 acquired immunodeficiency syndrome (AIDS)-related deaths in 2020. The only way to increase the life expectancy of these patients right now is to use combination antiretroviral therapy (cART) for the lifetime. Due to the integration of the HIV-1 DNA in lymphocytes, the replication of the virus can only be reduced by using antiretroviral drugs. If the drug is stopped, the virus will replicate and reduce the number of lymphocytes. In recent years, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease Cas9-mediated genome editing system has been considered, preventing HIV-1 replication by causing DNA double-stranded breaks (DSBs) or disrupting the integrated virus replication by targeting the provirus. In this study, we utilized the CRISPR/Cas9 without the nuclear localization signal sequence (w/o NLS) system to inhibit the VSV-G-pseudotyped HIV-1 replication by targeting the HIV-1 DNA as a prophylactic method. To this end, we designed a multiplex gRNA (guide RNA) cassette to target the

INTRODUCTION: Resistance to azole drugs has been observed in candidiasis due to their long‐term use and poor response to treatment. Resistance to azole drugs in Candida albicans isolates is controlled by several genes including ERG11, CDR1, CDR2, and MDR1. In this study, the expression of the mentioned genes was evaluated in C. albicans isolates susceptible and resistant to fluconazole. METHODS: After identifying the Candida isolates using morphological and molecular methods, the minimum inhibitory concentration (MIC) and drug susceptibility were determined using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) method. RNA was then extracted and cDNA was synthesized from 24 C. albicans isolates from patients with cancer. Then, the mean expressions of these genes were compared in two groups using real‐time polymerase chain reaction (RT‐PCR). RESULTS: A total of 74 Candida isolates were obtained from the oral cavity of 61 cancer patients with oral candidiasis. After 24 h, 21.6% of the isolates were fluconazole‐resistant, 10.8% were identified as dose‐dependent, and the rest of the isolates (67.6%) were fluconazole‐sensitive. The mean expressions of the CDR1 and MDR1 genes were significantly higher in the resistant isolates than in the sensitive ones. However, the ERG11 and CDR2 genes were not significantly increased in the resistant isolates. CONCLUSION: The increased mean expressions of the CDR1 and MDR1 genes had a greater effect on fluconazole resistance among the drug‐resistant strains of C. albicans in chemotherapy patients. It seemed that the accumulation of chemotherapeutic drugs in this organism stimulated some regulatory factors and increased the expression of these two genes and ultimately helped to further increase their expression and resistance to fluconazole.