Phase II randomized, double blind, placebo controlled, clinical trial of safety and immunogenicity of an inactivated SARS-CoV-2 vaccine FAKHRAVAC in adults aged 18-70 years.

BACKGROUND: The FAKHRAVAC®, an inactivated SARS-CoV-2 vaccine, was assessed for safety and immunogenicity in a phase II trial. METHODS: We did a phase II, single-centered, randomized, double-blind, placebo-controlled clinical trial of the FAKHRAVAC inactivated SARS-CoV-2 vaccine on adults aged 18 to 70. The two parallel groups received two intramuscular injections of either a 10-µg vaccine or a placebo at 2-week intervals. The participants' immunogenicity responses and the occurrence of solicited and unsolicited adverse events were compared over the study period of up to 6 months. Immunogenicity outcomes include serum neutralizing antibody activity and specific IgG antibody levels. RESULTS: Five hundred eligible participants were randomly (1:1) assigned to vaccine or placebo groups. The median age of the participants was 36 years, and 75% were male. The most frequent local adverse reaction was tenderness (21.29% after the first dose and 8.52% after the second dose), and the most frequent systemic adverse reaction was headache (11.24% after the first dose and 8.94% after the second dose). Neutralizing antibody titers two and four weeks after the second injection in the vaccine group showed about 3 and 6 times increase compared to the placebo group (GMR = 2.69, 95% CI 2.32-3.12, N:309) and (GMR = 5.51, 95% CI 3.94-8.35, N:285). A four-fold increase in the neutralizing antibody titer was seen in 69.6% and 73.4% of the participants in the vaccine group two and four weeks after the second dose, respectively. Specific ELIZA antibody response against a combination of S1 and RBD antigens 4 weeks after the second injection increased more than three times in the vaccine compared to the placebo group (GMR = 3.34, 95% CI 2.5-4.47, N:142). CONCLUSIONS: FAKHRAVAC® is safe and induces a significant humoral immune response to the SARS-CoV-2 virus at 10-µg antigen dose in adults aged 18-70. A phase III trial is needed to assess the clinical efficacy. TRIAL REGISTRATION: Trial Registry Number: Ref., IRCT20210206050259N2 ( http://irct.ir ; registered on 08/06/2021).

Physiological responses of plants and mites to salicylic acid improve the efficacy of spirodiclofen for controlling Tetranychus urticae (Acari: Tetranychidae) on greenhouse tomatoes.

Salicylic acid (SA) is a signaling molecule that can induce plant resistance to certain herbivores. Although the role of jasmonic acid in mediating mite-tomato plant interactions has been well studied, the role of salicylic acid has not. This study examined how the application of exogenous SA, via its effects on tomato plant physiology, alters the activity of mite digestive enzymes, mite energy reserves, and mite susceptibility to spirodiclofen. Enzymatic activity-including superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase-along with contents of total phenolic, hydrogen peroxide, and total chlorophyll significantly increased in plants 24 h after treatment with 2 mM of SA. In contrast, catalase activity significantly decreased in treated plants, and malondialdehyde content was unaffected. Mites fed on tomato plants treated with SA had significantly lower glutathione S-transferase, esterase, α-amylase, and aminopeptidase activities than those fed on control plants. Energy reserve analyses demonstrated a significant decrease in contents of lipid, protein, and glycogen in mites fed on SA-treated plants, whereas carbohydrate content significantly increased. The LC50 of spirodiclofen was decreased 1.8-fold for Tetranychus urticae fed on SA-treated tomato plants compared to controls. Treatment of adult mites with 2 mM SA on leaf discs did not cause any direct mortality after 24 h. Finally, a greenhouse bioassay confirmed that spider mite mortality following exposure to spirodiclofen was significantly higher on SA plants than on control plants. Mortality of mites exposed to half of the recommended rate of spirodiclofen was similar to those exposed to the recommended rate when they were held on treated plants. These results have valuable implications for T. urticae management programs in tomato production.

Antixenosis and antibiosis response of common bean (Phaseolus vulgaris) to two-spotted spider mite (Tetranychus urticae).

The two-spotted spider mite, Tetranychus uticae Koch (Acari: Tetranychidae), is globally one of the most devastating pests that feed on numerous crops, including common bean (Phaseolus vulgaris L.). This study was aimed to evaluate the effects of genotype and morphological attributes of common bean on T. uticae. Forty common bean accessions were used to investigate antixenosis and antibiosis through assessing mite feeding preference and reproduction under laboratory conditions. Three resistant (i.e., 56, 63, 238) and two susceptible (i.e., 182, 236) accessions, along with cultivars Naz (resistant) and Akhtar (susceptible), were used in a life-table study. Both antixenosis and antibiosis mechanism were observed in all of the accessions, albeit a negative correlation occurred. Significant differences were observed for all traits of T. urticae: developmental time of immature stages, reproduction, adult longevity and life-table parameters. Based on the intrinsic rate of increase, the accessions 56, 63, 182, 238, and cv. Naz impose high antibiotic effects on T. urticae. Although significant variation existed among accessions for morphological factors, only glandular trichomes correlated with mite fecundity and feeding preference.

Immunotoxin: A new tool for cancer therapy.

Cancer is one of the main reasons of death in the most countries and in Iran. Immunotherapy quickly became one of the best methods of cancer treatment, along with chemotherapy and radiation. "Immunotoxin Therapy" is a promising way of cancer therapy that is mentioned in this field. Immunotoxins are made from a toxin attaching to an antibody target proteins present on cancer cells. The first-generation immunotoxins were made of a full-length toxin attached to whole monoclonal antibodies. But, these immunotoxins could bind to normal cells. DAB389IL2 was the first immunotoxin approved by the Food and Drug Administration. Current trends and researches are ongoing on finding proteins that in combination with immunotoxins have minimal immunogenicity and the most potency for target cell killing.

Functional Response of Eretmocerus delhiensis on Trialeurodes vaporariorum by Parasitism and Host Feeding.

The parasitoid wasp, Eretmocerus delhiensis (Hymenoptera, Aphelinidae) is a thelytokous and syn-ovigenic parasitoid. To evaluate E. delhiensis as a biocontrol agent in greenhouse, the killing efficiency of this parasitoid by parasitism and host-feeding, were studied. Killing efficiency can be compared by estimation of functional response parameters. Laboratory experiments were performed in controllable conditions to evaluate the functional response of E. delhiensis at eight densities (2, 4, 8, 16, 32, 64, 100, and 120 third nymphal stage) of Trialeurodes vaporariorum (Hemiptera, Aleyrodidae) on two hosts including; tomato and prickly lettuce. The maximum likelihood estimates from regression logistic analysis revealed type II functional response for two host plants and the type of functional response was not affected by host plant. Roger's model was used to fit the data. The attack rate (a) for E. delhiensis was 0.0286 and 0.0144 per hour on tomato and 0.0434 and 0.0170 per hour on prickly lettuce for parasitism and host feeding, respectively. Furthermore, estimated handling times (Th) were 0.4911 and 1.4453 h on tomato and 0.5713 and 1.5001 h on prickly lettuce for parasitism and host feeding, respectively. Based on 95% confidence interval, functional response parameters were significantly different between the host plants solely in parasitism. Results of this study opens new insight in the host parasitoid interactions, subsequently needs further investigation before utilizing it for management and reduction of greenhouse whitefly.

Safety and immunogenicity of an inactivated SARS-CoV-2 vaccine (FAKHRAVAC®) in healthy adults aged 18-55 years: Randomized, double-blind, placebo-controlled, phase I clinical trial.

Background: The FAKHRAVAC®, an inactivated SARS-CoV-2 vaccine, was assessed for safety and immunogenicity. Methods and findings: In this double-blind, placebo-controlled, phase I trial, we randomly assigned 135 healthy adults between 18 and 55 to receive vaccine strengths of 5 or 10 µg/dose or placebo (adjuvant only) in 0-14 or 0-21 schedules. This trial was conducted in a single center in a community setting. The safety outcomes in this study were reactogenicity, local and systemic adverse reactions, abnormal laboratory findings, and Medically Attended Adverse Events (MAAE). Immunogenicity outcomes include serum neutralizing antibody activity and specific IgG antibody levels.The most frequent local adverse reaction was tenderness (28.9%), and the most frequent systemic adverse reaction was headache (9.6%). All adverse reactions were mild, occurred at a similar incidence in all six groups, and were resolved within a few days. In the 10-µg/dose vaccine group, the geometric mean ratio for neutralizing antibody titers at two weeks after the second injection compared to the placebo group was 9.03 (95% CI: 3.89-20.95) in the 0-14 schedule and 11.77 (95% CI: 2.77-49.94) in the 0-21 schedule. The corresponding figures for the 5-µg/dose group were 2.74 (1.2-6.28) and 5.2 (1.63-16.55). The highest seroconversion rate (four-fold increase) was related to the 10-µg/dose group (71% and 67% in the 0-14 and 0-21 schedules, respectively). Conclusions: FAKHRAVAC® is safe and induces a strong humoral immune response to the SARS-CoV-2 virus at 10-µg/dose vaccine strength in adults aged 18-55. This vaccine strength was used for further assessment in the phase II trial.Trial registrationThis study is registered with https://www.irct.ir; IRCT20210206050259N1.

Physiological Differences Between Seasonal Dimorphs of Agonoscena pistaciae (Hemiptera: Aphalaridae) Elicit Distinct Host Plant Responses, Informing Novel Pest Management Insights.

We examined differences in the physiology and life history between dimorphs of the common pistachio psyllid, Agonoscena pistaciae (Burckhardt and Lauterer) (Hemiptera: Aphalaridae), and how they differ in elicitating host plant production of key metabolites and volatile compounds involved in the recruitment of herbivores and natural enemies. Summer morphs had higher activities of glutathione S-transferase, carboxylesterase, acetylcholinesterase, and cytochrome P450 monooxygenase, superoxide dismutase, catalase, peroxidase, phenoloxidase, and a higher total protein content compared to winter morphs, whereas the latter had higher amounts of lipid, carbohydrate, and glycogen. Winter morphs were heavier, with a higher chitin content and longer preoviposition period, but greater fecundity and longevity than summer morphs. A lower LC50 to thiamethoxam for winter morphs resulted in higher mortality following exposure to the recommended rate of this insecticide in a greenhouse trial. Feeding by winter morphs elicited more strongly the release of volatile compounds known to be attractive to other herbivores, whereas feeding by summer morphs elicited more strongly the release of volatiles implicated in the attraction of natural enemies. Feeding by psyllids increased the concentrations of nitrogenous compounds, carbohydrates, vitamins, and amino acids in plants, the winter morph eliciting larger changes and more improved host plant quality. We conclude that winter morphs are more vulnerable targets for chemical control in early spring, whereas management of summer morphs could rely more on conservation biological control.

Ant mimicry by an aphid parasitoid, Lysiphlebus fabarum.

In Iran, Lysiphlebus fabarum (Marshall) (Hymenoptera: Braconidae: Aphidiinae) is a uniparental parasitoid of the black bean aphid, Aphis fabae Scopoli (Hemiptera: Aphididae), that possesses various highly evolved adaptations for foraging within ant-tended aphid colonies. Direct observations and video recordings were used to analyze the behavior of individual females foraging for A. fabae on bean leaf disks in open arenas in the laboratory. Females exploited aphids as hosts and as a source of food, allocating within-patch time as follows: resting - 10.4%, grooming - 8.2%, searching - 11.5%, antennation (host recognition) - 7.5%, antennation (honeydew solicitation mimicking ants) - 31.9%, abdominal bending (attack preparation) 19.7%, probing with the ovipositor (attack) - 10.8%. The mean handling time for each aphid encountered was 2.0 ± 0.5 min. Females encountered an average of 47.4 ± 6.4 aphids per hour, but laid only 1.2 eggs per hour. The ovipositor insertion time for parasitism ranged from 2 sec to longer than a minute, but most insertions did not result in an egg being laid. A. fabae defensive behaviors included kicking, raising and swiveling the body, and attempts to smear the attacker with cornicle secretions, sometimes with lethal results. Food deprivation for 4-6 h prior to testing increased the frequency of ant mimcry by L. fabarum. Females also used ant-like antennation to reduce A. fabae defensive behavior, e.g. the frequency of kicking. L. fabarum attacks primed A. fabae to be more responsive to subsequent honeydew solicitation, such that experienced females improved their feeding success by alternating between the roles of parasitoid and ant mimic. These results reveal the possibility for mutualisms to evolve between L. fabarum and the ant species that tend A. fabae, since L. fabarum receive ant protection for their progeny and may benefit the ants by improving A. fabae responsiveness to honeydew solicitation.

Micronutrient Fertilization of Greenhouse Cucumbers Mitigates Pirimicarb Resistance in Aphis gossypii (Hemiptera: Aphididae).

The nutritional status of host plants can have direct impacts on herbivore physiology and insect-plant interactions. We investigated the effect of micronutrients, including manganese, iron, zinc, and copper, on cucumber plant physiology, and on the biology and physiology of a strain of Aphis gossypii Glover selected over 12 generations to be resistant to pirimicarb. The micronutrient treatment increased the activity of superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase in cucumber plants, and also increased levels of total phenolics, hydrogen peroxide, salicylic acid, and total chlorophyl, whereas malondialdehyde levels were unaffected. Pirimicarb-resistant cotton aphids that fed on micronutritient-amended cucumber plants expressed significantly decreased levels of acetylcholinesterase and detoxifying enzymes, specifically glutathione S-transferase, and carboxylesterase. Analysis of energy reserves in resistant A. gossypii fed on micronutritient-amended plants revealed decreases in the lipid and protein contents of aphids, whereas glycogen and carbohydrate contents showed no response. Resistant cotton aphids fed on micronutritient-amended plants showed significantly reduced fecundity, longevity, and reproductive periods, and a 1.7-fold reduction in pirimicarb LC50 compared with those fed on control plants. We conclude that micronutrient amendment negatively impacts the biological performance of insecticide-resistant cotton aphids, and diminishes their resistance to pirimicarb. Both direct effects on plant health, such as enhanced inducible defenses, and indirect effects on aphid fitness, such as reduced biological performance and detoxification abilities, were implicated. Therefore, optimization of micronutrient amendments could be a useful complement to other tactics for managing insecticide-resistant A. gossypii on cucumbers, and warrants exploration in other contexts.

Comparative transcriptome provides molecular insight into defense-associated mechanisms against spider mite in resistant and susceptible common bean cultivars.

Common bean (Phaseolus vulgaris L.) is a major source of proteins and one of the most important edible foods for more than three hundred million people in the world. The common bean plants are frequently attacked by spider mite (Tetranychus urticae Koch), leading to a significant decrease in plant growth and economic performance. The use of resistant cultivars and the identification of the genes involved in plant-mite resistance are practical solutions to this problem. Hence, a comprehensive study of the molecular interactions between resistant and susceptible common bean cultivars and spider mite can shed light into the understanding of mechanisms and biological pathways of resistance. In this study, one resistant (Naz) and one susceptible (Akhtar) cultivars were selected for a transcriptome comparison at different time points (0, 1 and 5 days) after spider mite feeding. The comparison of cultivars in different time points revealed several key genes, which showed a change increase in transcript abundance via spider mite infestation. These included genes involved in flavonoid biosynthesis process; a conserved MYB-bHLH-WD40 (MBW) regulatory complex; transcription factors (TFs) TT2, TT8, TCP, Cys2/His2-type and C2H2-type zinc finger proteins; the ethylene response factors (ERFs) ERF1 and ERF9; genes related to metabolism of auxin and jasmonic acid (JA); pathogenesis-related (PR) proteins and heat shock proteins.

Circadian Rhythmicity of Diazinon Susceptibility, Detoxifying Enzymes, and Energy Reserves in Aphis gossypii (Hemiptera: Aphididae).

The daily susceptibility rhythm of the cotton aphid, Aphis gossypii Glover, to diazinon and the corresponding changes in the activity of three xenobiotic detoxifying enzymes-cytochrome P450 monooxygenases (P450), glutathione S-transferases (GSTs), and esterases-were investigated. Bioassays were conducted to estimate the median lethal doses (LD50) of diazinon at six different zeitgeber times (ZT0, 4, 8, 12, 16, and 20) under constant light (LL) and lighting conditions of 16 h of light and 8 h of darkness (LD). The results showed that the highest susceptibility occurred at the onset of night (ZT16) and 4 h before this time point (ZT12) under the LD condition. The endogenous rhythmicity of susceptibility was ensured, as the highest susceptibility occurred at the same time under the LL condition. The circadian changes in susceptibility to diazinon were almost coincident with changes in esterase and GSTs activity, but not in P450 activity. We also found rhythmic changes in energy components of whole-body aphids, with similar patterns of circadian changes of proteins, lipids, and soluble carbohydrates, but not glycogen, under LL and LD conditions. These photoperiod conditions (LD and LL) showed different fluctuation in trends of energy resources and of course, different quantities. Our study represents the first report of circadian control of insecticide susceptibility in aphids and provides insights into more efficient control of these pests by unveiling the times of day during which aphids are more susceptible to insecticides with attention to endogenous physiological phenomena.

Dengue virus type-3 envelope protein domain III; expression and immunogenicity.

OBJECTIVES: Production of a recombinant and immunogenic antigen using dengue virus type-3 envelope protein is a key point in dengue vaccine development and diagnostic researches. The goals of this study were providing a recombinant protein from dengue virus type-3 envelope protein and evaluation of its immunogenicity in mice. MATERIALS AND METHODS: Multiple amino acid sequences of different isolates of dengue virus type-3, corresponding to the envelope protein domain III, were achieved from GenBank. Clustal V alignment tool was used to provide a consensus amino acid sequence. Nucleotide sequence of the coding gene was optimized using "Optimizer". The origami (DE3) strain of Escherichia coli was used as the host in order to express the protein. A commercial affinity chromatography method was used to purify the recombinant protein. Immunogenicity of the recombinant protein was evaluated in mice using ELISA, MTT and cytokine assays. RESULTS: A consensus amino acid sequence corresponding to the most important region of dengue virus type-3 envelope protein (domain III) was provided. A high concentration (≥ 20 mg/L culture medium) of soluble recombinant antigen (EDIII3) was achieved. Immunized mice developed specific antibody responses against EDIII3 protein. The splenocytes from EDIII3-immunized mice showed a high proliferation rate in comparison with the negative control. In addition, the concentrations of two measured cytokines (IFN-γ and IL-4) were increased markedly in immunized mice. CONCLUSION: The results showed that the expressed recombinant EDIII3 protein is an immunogenic antigen and can be applied to induce specific immune responses against dengue virus type-3.

Functional response of larval and adult stages of Hippodamia variegata (Coleoptera: Coccinellidae) to different densities of Aphis fabae (Hemiptera: Aphididae).

We compare the efficiencies of different stages of Hippodamia variegata (Coleoptera: Coccinellidae) preying on Aphis fabae (Scolpoli) (Hemiptera: Aphididae) by estimating the functional responses of all stages. The experiments were carried out on leaf disks in petri dishes with 15-20 replicates. Our results revealed that all larval instars and adult males and females of H. variegata exhibited type II functional responses on different densities of prey. The rate of searching efficiency and handling time were estimated as 0.063 h(-1) and 6.933 h for first instar, 0.059 h(-1) and 3.343 h for second instar, 0.103 h(-1) and 1.909 h for third instar, 0.114 h(-1) and 0.455 h for fourth instar, 0.159 h(-1) and 1.194 h for male, 0.093 h(-1) and 0.409 h for female, respectively. Thus, handing time decreased from first instar to female. Handling times of males were significantly greater than those of females. The most effective stages of H. variegata were females, fourth instars, and males. The efficiency of females was nearly three times greater than that of males. The voracity of larval stages and male and female adults of H. variegata were estimated as 2.93, 5.85, 12.13, 45.13, 18.33, and 44.60 (aphids/d), respectively.

Effect of host plants on developmental time and life table parameters of Amphitetranychus viennensis (Acari: Tetranychidae).

The effect of host plant species including black cherry (Prunus serotina cv. Irani), cherry (Prunus avium cv. siahe Mashhad) and apple (Malus domestica cv. shafi Abadi) was studied on biological parameters of Amphitetranychus viennensis (Zacher) in the laboratory at 25 +/- 1 degrees C, 70 +/- 10% RH and 16L: 8D photoperiod. Duration of each life stage, longevity, reproduction rate, the intrinsic rate of natural increase (rm), net reproductive rate (R0), mean generation time (T), doubling time (DT), and finite rate of increase (lambda) of the hawthorn spider mite on the three host plants were calculated. Differences in fertility life table parameters of the spider mite among host plants were analyzed using pseudo-values, which were produced by jackknife re-sampling. The results indicated that black cherry might be the most suitable plant for hawthorn spider mite due to the shorter developmental period (10.6 days), longer adult longevity (25.5 days), higher reproduction (65.6 eggs), and intrinsic rate of natural increase (0.194 females/female/day). Cherry was the least suitable host plant. To determine the effect of host shifts, the mite was transferred from black cherry onto cherry and apple. In the first generation after shifting to apple, the developmental period, reproduction and life table parameters were negatively influenced. However, population growth parameters in the first generation on cherry were actually better than after three generations on this new host. This underscores the relevance of the mites' recent breeding history for life table studies.

Predation by Allothrombium pulvinum on the spider mites Tetranychus urticae and Amphitetranychus viennensis: predation rate, prey preference and functional response.

The deutonymphs of Allothrombium pulvinum Ewing (Acari: Trombidiidae) are among the most important natural enemies of spider mites in North, North East and West Iran. In this study, maximum predation rate and preference experiments were conducted with A. pulvinum deutonymphs on apple leaf discs, to determine their preference for either of two spider mite species: Amphitetranychus viennensis (Zacher) and Tetranychus urticae Koch (Acari: Tetranychidae). Maximum predation rate tests showed that the predatory mite consumed more eggs and females of T. urticae than of A. viennensis. Furthermore, the Manly's preference index for eggs and females of T. urticae confirmed that T. urticae were the preferred prey. The functional response of A. pulvinum deutonymphs on females of T. urticae was examined over a 24-h period. Predation of A. pulvinum deutonymphs presented with females of T. urticae followed a type III functional response. Estimated handling time for the predatory mites was 4.51 h and attack coefficient b, which describes the changes in attack rate with prey densities in a type III functional response, was 0.021.