Green Synthesis of Silver Nanoparticles Using Cirsium Congestum Extract Modified by Chitosan/Alginate: Bactericidal Activity against Pathogenic Bacteria and Cytotoxicity Analysis in Normal Cell Line.

AIM: The study aimed to determine in vitro pharmacological effects of modified Ag nanoparticles (AgNPs). BACKGROUND: AgNPs are considered antimicrobial agents. However, the cytotoxicity of chemically synthesized AgNPs (cAgNPs) has raised challenges that limit their use. OBJECTIVE: The purpose of the study was to examine the antimicrobial and cytotoxicity effects of AgNPs synthesized using Cirsium congestum extract modified by chitosan/alginate AgNPS (Ch/ALG-gAgNPs). METHODS: Nanoparticles were characterized using TEM, DLS, XRD, and FTIR. Resistant strains of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were used for the antimicrobial analysis of Ch/ALG-gAgNPs using disc diffusion and microdilution methods. The effects of NPs on cell viability and apoptosis in L929 normal cells were determined using MTT assay and annexin/PI staining, respectively. RESULTS: Physicochemical characterizations confirmed Ch/ALG-gAgNPs to be spherical and uniformly dispersed, and their size ranged from 50 to 500 nm. Ch/ALG-gAgNPs inhibited the growth of microbial strains in a dose-dependent manner. The antibacterial effect of Ch/ALG-gAgNPs was significantly higher than cAgNPs. The Ch/ALG-gAgNPs showed little cytotoxicity against normal cells at concentrations less than 50 µg/ml. Cytotoxicity effects of Ch/ALG-gAgNP were less than cAgNPs. Flow cytometry and real-time PCR results showed a decrease in apoptosis percentage and BAX marker in the presence of Ch/ALG-gAgNPs relative to when the cell was treated with cAgNPs. CONCLUSION: Current findings introduce novel gAgNPs modified with chitosan/alginate for use in medicine.

Molecular examination for Coxiella burnetii and Brucella spp. infections in Iranian women experiencing spontaneous miscarriage.

BACKGROUND: Spontaneous miscarriage, a leading health concern globally, often occurs due to various factors, including infections. Among these, Coxiella burnetii and Brucella spp. may have adverse effects on pregnancy outcomes. While previous research has established a link between infections and spontaneous miscarriage, our study aimed specifically to investigate the presence of these two pathogens in abortion samples from women who experienced spontaneous miscarriages in Iran. Our study can add to the existing knowledge by focusing on Iran, a region with a high prevalence of C. burnetii and Brucella spp. As a result, it could provide a better understanding and unique insights into the relationship of these pathogens with spontaneous miscarriages in endemic regions. METHODS: From March 2021 to March 2022, a total of 728 abortion samples (including placenta and cotyledon) were collected from 409 women who had experienced spontaneous miscarriages in the provinces of Tehran, Fars, and West Azerbaijan in Iran. The specimens included 467 Formalin-Fixed Paraffin-Embedded (FFPE) and 261 fresh frozen samples. After DNA extraction from abortion samples, the quantitative real-time PCR (qPCR) assay targeted a specific fragment of the IS1111 and IS711 elements for molecular identification of C. burnetii and Brucella spp., respectively. Furthermore, the qPCR assay employing specific primers for different species was used to determine the species of Brucella. RESULTS: Among the studied women, 1 out of 409 (0.24%) samples tested positive for Brucella spp., specifically Brucella melitensis. There were no positive specimens for C. burnetii. CONCLUSIONS: Our study contributes to understanding the potential involvement of Brucella species in spontaneous infectious abortion within endemic regions. The identification of B. melitensis in this study highlights the need for further research in this area. However, while our results suggest a relatively low or zero identification of these pathogens in our sample population, this does not rule out the possibility of undetected infections. Therefore, it is critical to acknowledge the limitations of the molecular techniques used (qPCR), which may have potential limitations such as sensitivity and specificity. Moreover, because 64.15% of our samples were FFPE, the sensitivity of the qPCR test may be reduced. These raise concerns about the accuracy of the reported prevalence rates and the potential for false positives or negatives.

Efficacy of nutritional selenium nanoparticles on growth performance, immune response, antioxidant capacity, expression of growth and immune-related genes, and post-stress recovery in juvenile Sobaity seabream (Sparidentex hasta).

This study evaluated the impacts of nano-Se on the growth, immunity, antioxidant capacity, physiological parameters, gene expression, and stress resistance of fingerling Sobaity seabream (Sparidentex hasta). The fish with an average weight of 21.5 ± 0.1 g were divided into four treatment groups in triplicates that received one of the test diets supplemented with varying levels of nano-Se: 0 (control), 0.5 (Se-0.5), 1 (Se-1), and 2 (Se-2) mg/Kg for 60 days. The results showed that final weight, weight gain rate, specific growth rate, feed intake, and feed conversion ratio improved with significant linear and quadratic trends (P < 0.05) in response to nano-Se-supplemented diets, and the best values were measured in the Se-2 group. Superoxide dismutase activity level remained unaffected among the four groups (P > 0.05). Catalase activity increased in nano-Se-supplemented groups, with the highest level measured in fish fed the Se-0.5 diet. Glutathione peroxidase activity levels were not significantly different between the control and nano-Se groups, but the lowest malondialdehyde concentration was detected in the Se-2 group. Nano-Se had no marked effect on total plasma Ig levels; however, the highest lysozyme activity and alternative complement activity (ACH50) were observed in the Se-0.5 and Se-2 groups, respectively. No significant differences (P > 0.05) were observed in plasma total protein, albumin, globulin, triglyceride, and thyroid hormone (T3 and T4) contents among the groups. However, the lowest cholesterol and low-density lipoprotein values and the highest high-density lipoprotein concentration were measured in the Se-2 group. The Se-0.5 and Se-1 groups exhibited significantly lower levels of aspartate aminotransferase activity, and the lowest alkaline phosphatase activity level was detected in the Se-1 group. The expression level of insulin-like growth factor I gene in all nano-Se-fed groups was significantly higher than the control. Also, the expression of interleukin-1ß and lysozyme genes was significantly upregulated in nano-Se-supplemented groups, with the highest values in the Se-2 group. Following acute crowding stress, plasma cortisol and lactate levels at all post-stress time intervals were not significantly different among the experimental groups. Fish fed the Se-0.5 and Se-2 diets tended to have lower plasma glucose concentrations than other groups. In conclusion, dietary nano-Se at 2 mg/kg is recommended to promote growth performance and enhance antioxidant and immune parameters in Sobaity juveniles.

The surveillance of plague among rodents and dogs in Western Iran.

BACKGROUND: The causative agent of plague, Yersinia pestis, is maintained in nature via a flea-rodent cycle. Western Iran is an old focus for plague, and recent data indicate that rodents and dogs in this region have serological evidence of Y. pestis infection. The purpose of this study was to conduct a large-scale investigation of Y. pestis infection in shepherd dogs, rodents, and their fleas in old foci for plague in Western Iran. MATERIALS AND METHODS: This study was conducted in Hamadan province from 2014 to 2020. Rodents and fleas were collected from various locations throughout this region. Y. pestis was investigated in rodent spleen samples and fleas using culture, serology, and real-time PCR methods. Additionally, sera samples were collected from carnivores and hares in this region, and the IgG antibody against the Y. pestis F1 antigen was assessed using an ELISA. RESULTS: In this study, 927 rodents were captured, with Meriones spp. (91.8%) and Microtus qazvinensis (2.6%) being the most prevalent. A total of 6051 fleas were collected from rodents and carnivores, most of which were isolated from Meriones persicus. None of the rodents or fleas examined tested positive for Y. pestis using real-time PCR and culture methods. Meanwhile, IgG antibodies were detected in 0.32% of rodents. All serologically positive rodents belonged to M. persicus. Furthermore, none of the sera from the 138 carnivores (129 sheepdogs, five Vulpes vulpes, four Canis aureus), and nine hares tested positive in the ELISA test. CONCLUSION: This primary survey of rodent reservoirs shows serological evidence of Y. pestis infection. Western Iran is an endemic plague focus, and as such, it requires ongoing surveillance.

The significance of essential oils and their antifungal properties in the food industry: A systematic review.

Essential oils (EOs) are natural products called volatile oils or aromatic and ethereal oils derived from various parts of plants. They possess antioxidant and antimicrobial properties, which offer natural protection against a variety of pathogens and spoilage microorganisms. Studies conducted in the last decade have demonstrated the unique applications of these compounds in the fields of the food industry, agriculture, and skin health. This systematic article provides a summary of recent data pertaining to the effectiveness of EOs and their constituents in combating fungal pathogens through diverse mechanisms. Antifungal investigations involving EOs were conducted on multiple academic platforms, including Google Scholar, Science Direct, Elsevier, Springer, Scopus, and PubMed, spanning from April 2000 to October 2023. Various combinations of keywords, such as "essential oil," "volatile oils," "antifungal," and "Aspergillus species," were used in the search. Numerous essential oils have demonstrated both in vitro and in vivo antifungal activity against different species of Aspergillus, including A. niger, A. flavus, A. parasiticus, A. fumigatus, and A. ochraceus. They have also exhibited efficacy against other fungal species, such as Penicillium species, Cladosporium, and Alternaria. The findings of this study offer novel insights into inhibitory pathways and suggest the potential of essential oils as promising agents with antifungal and anti-mycotoxigenic properties. These properties could make them viable alternatives to conventional preservatives, thereby enhancing the shelf life of various food products.

Molecular investigation of Coxiella burnetii and Francisella tularensis infection in ticks in northern, western, and northwestern Iran.

Tularemia and Q fever are endemic diseases in Iran; however, little information is available on the prevalence of the causative agents, Coxiella burnetii and Francisella tularensis, in Iranian ticks. This study investigated C. burnetii and F. tularensis among hard ticks in this country. We collected ticks from livestock and other mammals in Guilan, Mazandaran, Golestan (northern Iran), Kurdistan (western Iran), and West Azerbaijan (northwestern Iran) provinces. Genomic DNA from collected ticks was extracted and screened for C. burnetii and F. tularensis using Real-time PCR. A total of 4,197 ticks (belonging to 12 different species) were collected, and Ixodes ricinus (46.4%), Rhipicephalus turanicus (25%), and Rhipicephalus sanguineus sensu lato (19.1%) were the most collected species. Of 708 pooled tick samples, 11.3% and 7.20% were positive for C. burnetii and F. tularensis, respectively. The genus of Rhipicephalus had the highest (18.3%) C. burnetii infection among the collected tick pools (P<0.001). Furthermore, the most positive pools for F. tularensis belonged to Haemaphysalis spp. (44.4%). Kurdistan had the most significant percentage of C. burnetii-infected ticks (92.5%), and there was a meaningful relationship between the provinces and the infection (P< 0.001). The ticks from Golestan exhibited the highest F. tularensis infection rate (10. 9%), and the infection showed no significant relationship with the provinces (P = 0.19). Ticks collected from grasslands had a higher Coxiella burnetii infection rate than those collected from animals (39.4% vs. 7.9%; p<0.01). However, ticks collected from animal surfaces had a slightly higher rate of Francisella tularensis infection than those collected from grasslands (7.6% vs. 3.9%; p = 0.24). Here, we demonstrated the presence of both pathogens in the north (Guilan, Mazandaran, and Golestan provinces), the west (Kurdistan province), and the northwest (West Azerbaijan province) of Iran. The public health system should pay particular attention to tick bites in veterinary medicine and humans.

Inhibitory activity of Limosilactobacillus reuteri isolated from camel milk against Helicobacter pylori effects in human gastric epithelial cells.

This study aimed to evaluate anti-Helicobacter pylori effects of Limosilactobacillus reuteri 2892 (L. reuteri 2892) isolated from camel milk in GC cell lines (AGS and MKN). From 15 camel milk samples, 132 microbial strains were isolated. Based on microbial and biochemical analysis, 11 potential probiotic candidates were selected. The potential probiotic candidates were assayed for anti-H. pylori activity, and the strain with the highest anti-H. pylori activity was identified genotypically. Based on 16S rDNA sequencing, the selected strain with the best activity against H. pylori (inhibition zone = 15.5 ± 0.8) belonged to the Lactobacillus reuteri strain 2892. Cell treatment with H. pylori HC-113 inhibits gene expression of Claudin-4, ZO-1, MUC5AC, and MUC2 in gastric cells, which are attenuated by L. reuteri 2892. The simulative effects of H. pylori HC-113 on the cell migration and invasion of gastric cells were lost when cells were cotreated with L. reuteri 2892. Cell treatment with H. pylori HC-113 promoted cell death, whereas cotreatment with L. reuteri 2892 markedly decreased necrotic and late apoptotic cells. The present study demonstrates that L. reuteri 2892 has potent anti-H. pylori effects and thus can be considered as an alternative protective agent against inflammatory effects of H. pylori in gastric cells.

The Anti-Helicobacter pylori effects of Limosilactobacillus reuteri strain 2892 isolated from Camel milk in C57BL/6 mice.

Helicobacter pylori infection (H. pylori) is associated with chronic gastritis, peptic ulcers, and gastric cancer. The present study provides information on the protective effects of Limosilactobacillus reuteri strain 2892 (L. reuteri 2892) isolated from camel's milk against H. pylori-induced gastritis in the stomach tissue of animal models. Animal assays revealed that L. reuteri 2892 pretreatment significantly downregulated the virulence factor cagA gene expression. It upregulated the expression level of tight junction molecules [zona occludens (ZO-1), claudin-4] and suppressed metalloproteinase (MMP)-2 and MMP-9 expressions. L. reuteri 2892 exhibited immunomodulatory effects on cytokine profile, as it reduced the serum concentrations of pro-inflammatory cytokines interleukin (IL)-6, IL-1ß, and INF-γ and increased the anti-inflammatory cytokine, IL-10. In addition, L. reuteri 2892 showed anti-oxidative stress activity by regulating the levels of oxidative stress-associated markers [superoxide dismutase (SOD) and malondialdehyde (MDA)]. Our findings suggest that L. reuteri 2892 attenuates H. pylori-induced gastritis.

Green Synthesis of Silver Nanoparticles Using Salvadora persica and Caccinia macranthera Extracts: Cytotoxicity Analysis and Antimicrobial Activity Against Antibiotic-Resistant Bacteria.

Silver nanoparticles (AgNPs) have gained great interest because of their specific and distinct properties. Chemically synthesized AgNPs (cAgNPs) are often unsuitable for medical applications due to requiring toxic and hazardous solvents. Thus, green synthesis of AgNPs (gAgNPs) using safe and nontoxic substances has attracted particular focus. The current study investigated the potential of Salvadora persica and Caccinia macranthera extracts in the synthesis of CmNPs and SpNPs, respectively. Aqueous extracts of Salvadora persica and Caccinia macranthera were prepared and taken as reducing and stabilizing agents through gAgNPs synthesis. The antimicrobial effects of gAgNPs against susceptible and antibiotic-resistant bacterial strains and their toxicity effects on L929 fibroblast normal cells were evaluated. TEM images and particle size distribution analysis showed that the CmNPs and SpNPs have average sizes of 14.8 nm and 39.4 nm, respectively. The XRD confirms the crystalline nature and purity of both CmNPs and SpNPs. FTIR results demonstrate the involvement of the biologically active substances of both plant extracts in the green synthesis of AgNPs. According to MIC and MBC results, higher antimicrobial effects were seen for CmNPs with a smaller size than SpNPs. In addition, CmNPs and SpNPs were much less cytotoxic when examined against a normal cell relative to cAgNPs. Based on high efficacy in controlling antibiotic-resistant pathogens without detrimental adverse effects, CmNPs may have the capacity to be used in medicine as imaging, drug carrier, and antibacterial and anticancer agents.

Serological evidence of Yersinia pestis infection in rodents and carnivores in Northwestern Iran.

BACKGROUND: Plague may recur after several decades in its endemic regions; therefore, the continuous monitoring of wildlife is essential, even when no human cases are reported in the old foci. The present study was conducted to monitor rodents and their ectoparasites as well as carnivores to learn about the epidemiology of plague infection in an old focus of Iran. METHODOLOGY: The present study was conducted from 2019 to 2020 in Takestan county of Qazvin Province in northwestern Iran. Rodents were caught using live traps, and their fleas were separated. Blood and spleen specimens were taken from the captured rodents. Serum samples were also collected from sheepdogs and wild carnivores. The collected samples were tested by culture, serology (ELISA), and molecular methods to detect Yersinia pestis infection. FINDINGS: A total of 399 small mammals were caught, of which 68.6% were Meriones persicus. A total of 2438 fleas were collected from the rodents, 95.3% of which were Xenopsylla buxtoni. Overall, 23 out of 377 tested rodents (5.7%, CI 95%, 3.9-9.0) had IgG antibodies against the F1 antigen of Y. pestis, and all the positive samples belonged to M. persicus. Nine (4.8%) out of 186 collected sera from the sheepdogs' serum and one serum from the Canis aureus had specific IgG antibodies against the F1 antigen of Y. pestis. There were no positive cases of Y. pestis in the rodents and fleas based on the culture and real-time PCR. CONCLUSION: Serological evidence of Y. pestis circulation was observed in rodents and carnivores (sheepdogs and C. aureus). The presence of potential plague vectors and serological evidence of Y. pestis infection in the surveyed animals could probably raise the risk of infection and clinical cases of plague in the studied region. Training health personnel is therefore essential to encourage their detection of possible human cases of the disease.

Myrtenol Ameliorates Recognition Memories' Impairment and Anxiety-Like Behaviors Induced by Asthma by Mitigating Hippocampal Inflammation and Oxidative Stress in Rats.

INTRODUCTION: Asthma is related to neurochemical alterations which affect brain functions and lead to anxiety and cognitive dysfunctions. Myrtenol has sparked considerable interest due to its pharmacological effects, especially for the remediation of chronic disorders. Thus, the present research was designed to evaluate the impacts of myrtenol on anxiety-like behaviors, cognitive declines, inflammation, and oxidative stress in the hippocampus of asthmatic rats. METHODS: Rats were allocated to five groups: control, asthma, asthma/vehicle, asthma/myrtenol, and asthma/budesonide. Asthma was elicited in the rats by ovalbumin, and the animals were then exposed to myrtenol inhalation. Anxiety-like behavior and memory were assessed by elevated plus maze (EPM) and novel object and location recognition tests. Interleukins (interleukin-6, -17, and -10), tumor necrosis factor α (TNF-α), and oxidative stress biomarkers such as malondialdehyde (MDA), superoxide dismutase (SOD), Glutathione peroxidase (GPX), and total antioxidant capacity (TAC) in the hippocampus were assessed by the ELISA method. RESULTS: The levels of IL-6, IL-17, TNF-α, and MDA decreased, but GPX, SOD, and TAC levels increased in the hippocampus of asthmatic animals due to myrtenol inhalation. CONCLUSION: Myrtenol diminished asthma-induced anxiety-like behaviors and cognitive deficits in asthmatic rats; these effects might have been typically mediated by a reduction in inflammation and oxidative stress.

Morphine upregulates Toll-like receptor 4 expression and promotes melanomas in mice.

BACKGROUND: Morphine and other opioids are used to manage cancer-related pain; however, the role of these drugs in cancer progression remains controversial. Emerging evidence indicates that morphine can activate Toll-like receptor 4 (TLR4) and its signaling pathways, by the way the activation and expression of TLR4 can promote melanoma. In this study, we investigated the effects of morphine on the expression of TLR4 and promotion of melanoma in mice. METHODS: Mice melanoma cells (B16F10) were cultured with morphine (0.1, 1 and 10 µM) for 24 h. In the other experiment, cells were treated with morphine with or without TLR4 agonist (LPS) or antagonist (TAK-242). In in-vivo model, B16F10 cells were subcutaneously injected to C57BL/6 mice, and morphine was administrated in three different treatment protocols after developing palpable tumors (acute treatment, chronic daily injections, escalating doses of morphine). In another set of experiments, B16F10 cells were pretreated with LPS (5 µg/ml) 24 h before injection into mice. Control group received normal saline. We measured cell proliferation, the expression level of Tlr4, Nuclear factor kappa-light-chain-enhancer of activated B cells 1 (Nf-κb1) genes, TLR4 protein expression, and tumor volume. RESULTS: Chronic, acute, and escalating doses of morphine increased tumor. Morphine increased the expression of Tlr4 and Nf-κb1 regardless of the treatment protocol used. CONCLUSION: Morphine increases the progression of melanoma cancer and may be related to the increased expression of TLR4. Our results suggest that morphine should be used with caution in patients with melanoma.HighlightsMorphine increases the expression of TLR4 in melanoma.Morphine increases melanoma progression.These effects are mostly observed with chronic and escalating morphine administration.

Morphine promotes migration and lung metastasis of mouse melanoma cells.

BACKGROUND: Morphine is an analgesic agent used for cancer pain management. There have been recent concerns that the immunosuppressant properties of morphine can also promote cancer metastasis. Morphine is an agonist for toll like receptor 4 (TLR4) that has a dual role in cancer development. The promotor or inhibitor role of morphine in cancer progression remains controversial. We investigated the effects of morphine on migration and metastasis of melanoma cells through TLR4 activation. METHODS: Mouse melanoma cells (B16F10) were treated with only morphine (0, 0.1, 1, and 10 µM) or in combination with a TLR4 inhibitor (morphine10 µM +CLI-095 1µM) for either 12 or 24 hours. Migration of cells was analyzed by transwell migration assays. Twenty C57BL/6 male mice were inoculated with B16F10 cells via the left ventricle of the heart and then randomly divided into two groups (n = 10 each) that received either morphine (10 mg.kg-1, sub-q) or PBS injection for 21 days (control group). Animals were euthanized and their lungs removed for evaluation of metastatic nodules. RESULTS: Morphine (0.1, 1, and 10 µM) increased cell migration after 12 hours (p < 0.001) and after 24 hours of treatment with morphine (10 µM) (p < 0.001). Treatment with CLI-095 suppressed migration compared to cells treated with morphine alone (p < 0.001). Metastatic nodules in the morphine-treated group (64 nodules) were significantly higher than in the control group (40 nodules) (p < 0.05). CONCLUSION: Morphine increases the migration and metastasis of mouse melanoma cells by activating TLR4.

The Combined Effects of Propionic Acid and a Mixture of Bacillus spp. Probiotic in a Plant Protein-Rich Diet on Growth, Digestive Enzyme Activities, Antioxidant Capacity, and Immune-Related Genes mRNA Transcript Abundance in Lates calcarifer Fry.

A 7-week feeding trial was conducted to evaluate the combined effects of propionic acid (PA, 5 or 10 g/kg) and a multi-strain Bacillus spp. (Bacillus subtilis IS02 (accession no. JN856456) and B. licheniformis IBRC-M 11,319) (1.7 × 107 CFU/g) probiotic in a plant protein source (PP)-rich diet (∼70% of dietary protein derived from PP sources) on performance of Asian sea bass (Lates calcarifer) fry (initial body weight 2.97 ± 0.11 g). In this regard, six isoproteic (∼48%) diets were formulated as follows: a control (without supplementation of the additives); probiotic (only contained Bacillus spp. mixture); 5 g PA/kg diet; 10 g PA/kg diet; probiotic + 5 g PA/kg diet, and probiotic + 10 g PA/kg diet. Specific growth rate in fish fed with 10 g PA/kg (2.84 ± 0.1%) and diets contained blends of probiotic and PA (2.76 ± 0.19% in probiotic + 5 g PA, and 2.79 ± 0.04% in probiotic + 10 g PA) was better than in the control (2.45 ± 0.1%) (P < 0.05). Feed conversion ratio in fish fed with 10 g PA/kg (0.92 ± 0.12) and diets contained blends of probiotic and PA (0.94 ± 0.06 in probiotic + 5 g PA and 0.91 ± 0.02 in probiotic + 10 g PA) was better than in the control (1.24 ± 0.08) (P < 0.05). Digestive enzymes including α-amylase, total alkaline proteases, and bile salt dependent lipase activities improved in fish fed diets contained additives. The activity of glutathione-S-transferase and glutathione reductase enhanced in the liver of fish fed diets contained additives. The relative abundance of lysozyme, interleukin 1ß, and insulin-like growth factor-1 genes mRNA transcript showed multifold increase in the liver of fish fed with the 10 g PA/kg and diets contained blends of probiotic and PA (P < 0.05). By considering the above mentioned results, supplementing a PP-rich diet with 10 g PA/kg diet or combination of PA and a mixture of Bacillus spp. probiotic recommended for L. calcarifer.

The effects of morphine on vascular cell adhesion molecule 1(VCAM-1) concentration in lung cancer cells.

OBJECTIVE: Vascular cell adhesion molecule 1 (VCAM-1) plays an important role in tumour cell adhesion to endothelial cells. Some tumour cells also show aberrant expression of VCAM-1. Toll-like receptor 4 (TLR4) agonists can increase VCAM-1 expression. Morphine, an opioid receptor agonist, is also a TLR4 agonist. In this study, we aimed to evaluate whether morphine increase VCAM-1 expression in a TLR4 dependent manner. METHODS: A549 Lung cancer cells were treated with different doses of morphine and TLR4 antagonist for 24 and 48 h. TLR4 gene expression was evaluated by real-time PCR and VCAM-1 protein was measured by the enzyme-linked immunosorbent assay (ELISA). RESULTS: Morphine enhanced mRNA expression of TLR4 and protein level of VCAM-1. TLR4 antagonist returned VCAM-1expression to the normal level. CONCLUSION: Morphine effects VCAM-1expressions via TLR4 in lung cancer cell line.

Morphine promotes migration and lung metastasis of mouse melanoma cells

Abstract Background Morphine is an analgesic agent used for cancer pain management. There have been recent concerns that the immunosuppressant properties of morphine can also promote cancer metastasis. Morphine is an agonist for toll like receptor 4 (TLR4) that has a dual role in cancer development. The promotor or inhibitor role of morphine in cancer progression remains controversial. We investigated the effects of morphine on migration and metastasis of melanoma cells through TLR4 activation. Methods Mouse melanoma cells (B16F10) were treated with only morphine (0, 0.1, 1, and 10 μM) or in combination with a TLR4 inhibitor (morphine10 μM +CLI-095 1μM) for either 12 or 24 hours. Migration of cells was analyzed by transwell migration assays. Twenty C57BL/6 male mice were inoculated with B16F10 cells via the left ventricle of the heart and then randomly divided into two groups (n = 10 each) that received either morphine (10 mg.kg−1, sub-q) or PBS injection for 21 days (control group). Animals were euthanized and their lungs removed for evaluation of metastatic nodules. Results Morphine (0.1, 1, and 10 μM) increased cell migration after 12 hours (p < 0.001) and after 24 hours of treatment with morphine (10 μM) (p < 0.001). Treatment with CLI-095 suppressed migration compared to cells treated with morphine alone (p < 0.001). Metastatic nodules in the morphine-treated group (64 nodules) were significantly higher than in the control group (40 nodules) (p < 0.05). Conclusion Morphine increases the migration and metastasis of mouse melanoma cells by activating TLR4.

Molecular surveillance for Rickettsia spp. and Bartonella spp. in ticks from Northern Iran.

Tick-borne zoonotic diseases pose a threat to public health; hence, identifying the pathogenic agents associated with them is critical. The prevalence of Bartonella and Rickettsia in Iran is unknown. This study aimed to detect Rickettsia spp. and Bartonella species in ticks in northeast Iran and conduct phylogenetic analysis on these bacteria. Ticks from the sample bank in the Research Center for Emerging and Re-emerging Diseases were included in this study. The ticks were collected in 2017 and 2018 from domestic animals (sheep, goats, cows, camels, horses, dogs, and donkeys) and rodents in Golestan, Mazandaran, and Guilan provinces. Molecular methods were used to examine the DNA extracted from these samples to detect Rickettsia spp. and Bartonella species. The study examined a total of 3999 ticks. Ixodes ricinus (46.4%), Rhipicephalus turanicus (26.3%), and Rhipicephalus sanguineus (17.1%) were the most prevalent species. Among 638 DNA pools, real-time-PCR detected Rickettsia spp. in 161 (25.2%), mostly belonging to Rh. sanguineus (48.9%) and Rh. turanicus (41.9%). Golestan Province had the highest number of positive pools (29.7%). No positive samples for Bartonella were detected in a 638 pooled samples. Eight distinct Rickettsia species were detected in 65 sequenced samples, the majority of which were R. massiliae (n = 32, 49.2%) and R. sibirica (n = 20, 30.8%). Other species included R. rhipicephali (n = 3), R. aeschlimannii (n = 5), R. helvetica (n = 5), R. asiatica (n = 4), R. monacensis (n = 6), and R. raoultii (n = 1). The research findings may provide helpful information about tick-borne Rickettsiae in Iran and help to clarify the role of these arthropods in maintaining these agents. Rickettsia species were found to be circulating in three Northern provinces; thus, it is recommended that this disease be considered in the differential diagnosis of febrile diseases caused by tick bites and febrile diseases with skin rashes such as Crimean-Congo hemorrhagic fever (CCHF).

The Inhibitory Effects of Vanillin on the Growth of Melanoma by Reducing Nuclear Factor-κB Activation.

Background: Melanoma is skin cancer, and the treatments are not efficient enough. Therefore, finding new drugs seems to be an essential need. Vanillin, which is extracted from vanilla seed, has anti-cancer effects by reducing nuclear factor-κB (NF). We explored the anti-tumor effects of vanillin in the melanoma model and its possible mechanism. Materials and Methods: In the MTT assay, mice melanoma cells (B16F10) were treated with vanillin (1, 2, 3, 4, 5 µg/mL) for 24 and 48 h. In an animal model, B16F10 was subcutaneously injected into C57BL/6 mice. After the development of tumors, the mice were treated with 50 and 100 mg/kg/day of vanillin for 10 days. The tumor size and expression level of NF-κB protein were measured. Results: In the MTT assay, vanillin in all concentrations significantly decreased B16F10 cell viability after 24 h incubation. The size of melanoma tumors was reduced in both doses 50 and 100 mg/kg/day in mice. NF-κB protein expression was decreased in the 100 mg/kg/day group in comparison with the control group. Conclusion: We found that vanillin by reducing NF-κB expression may have anti-tumor effects and reduced melanoma tumor size and cell viability.

Genetic species identification and population structure of grouper Epinephelus coioides (Hamilton, 1822) collected from fish markets along the Persian Gulf and the Oman Sea.

Many ecologically important and valuable fisheries marine species have been misidentified in terms of both the statistical data and market demand. Correct identification at the species level and the population genetic structure of the orange-spotted grouper (Epinephelus coioides), a precious fish in the Persian Gulf and the Oman Sea, was tested using mitochondrial cytochrome oxidase subunit I (DNA barcoding) and D-loop sequencing. The results revealed that the Epinephelus species found in the region, including E. coioides, E. bleekeri, E. polylepis, and E. chlorostigma were all mistakenly grouped together and identified as only E. coioides. Moreover, the analysis of molecular variance (AMOVA) of E. coioides samples using the D-loop showed a significantly unique genetic structure (ΦST = 0.068, p < 0.001) within the E. coioides population throughout the Persian Gulf and the Oman Sea, with the pairwise genetic difference between sampling locations in UAE and the Iranian coast. Moreover, D-loop sequences analysis showed two distinct haplotype groups scattered among the sampling locations, which did not correlate with the geographic distance between the sampling locations. These findings indicate that the issue of misidentification should be highlighted in the management and conservation of E. coioides. As this type of misidentification is likely to happen to other threatened marine species as well, the efficacy of using genetic markers for the correct identification, both at the species and the population level, is vital.

Metabolic Syndrome and Body Composition Among People Aged 50 Years and Over: Results from The Neyshabur Longitudinal Study on Ageing (NeLSA).

There are few studies regarding body composition and metabolic syndrome (MetS) association in older adults. To evaluate the association between MetS and body composition indices in a large-scale population of subjects with an age of 50 and up. This study was based on the data from Neyshabur Longitudinal Study on Ageing (NeLSA) in a total of 7462 people of Neyshabur city in IRAN. The best cut-off scores and AUC value of body composition variables for having association with likelihood of MetS were determined by using a receiver operating curve analysis. Each unit increase in the Waist/Hip ratio, the odds of having MetS increase 3-6 times (OR: 4.937, 95%CI: 3.930, 6.203 in men; OR: 3.322, 95%CI: 2.259, 4.884 in women). In addition, in the case of BMI (OR: 1.256, 95% Cl: 1.226, 1.286 in men; OR: 1.104, 95% Cl: 1.086, 1.121 in women) and BFM (OR: 1.119, 95% Cl: 1.105, 1.133 in men; OR: 1.050, 95% Cl: 1.041, 1.060 in women), the chance of having MetS increases with increasing these variables. Totally, BMI and BFM showed the best AUC values. The optimal cut-off values for BMI in men was 26.45 and in women was 27.35 and for BFM in men was 23.35 and in women was 26.85. These results suggest that adiposity measures such as BMI and BFM are associated with likelihood of having MetS in subjects with an age of 50 and up, and that avoiding high adiposity is important to prevent MetS incidence.