Generation and characterisation of P. falciparum parasites with a G358S mutation in the PfATP4 Na+ pump and clinically relevant levels of resistance to some PfATP4 inhibitors

Small-molecule inhibitors of PfATP4, a Plasmodium falciparum protein that is believed to pump Na+ out of the parasite while importing H+, are on track to become much-needed new antimalarial drugs. The spiroindolone cipargamin is poised to become the first PfATP4 inhibitor to reach the field, having performed strongly in Phase 1 and 2 clinical trials. Previous attempts to generate cipargamin-resistant parasites in the laboratory have yielded parasites with reduced susceptibility to the drug; however, the highest 50% inhibitory concentration reported to date is 24 nM. Here, we show that P. falciparum parasites can acquire a clinically-significant level of resistance to cipargamin that enables them to withstand micromolar concentrations of the drug. Independent experiments to generate high-level cipargamin resistance using different protocols and strains led to the same change each time - a G358S mutation in PfATP4. Parasites with this mutation showed high-level resistance not only to cipargamin, but also to the dihydroisoquinolone (+)-SJ733. However, for certain other (less clinically advanced) PfATP4-associated compounds the G358S mutation in PfATP4 conferred only moderate resistance or no resistance. The G358S mutation in PfATP4 did not affect parasite susceptibility to antimalarials that do not target PfATP4. The G358S mutation in PfATP4, and the equivalent mutation in the Toxoplasma gondii ATP4 homologue (G419S), decreased the sensitivity of the Na+-ATPase activity of ATP4 to inhibition by cipargamin and (+)-SJ733, and decreased the sensitivity of parasites expressing these ATP4 mutations to disruption of parasite Na+ regulation by cipargamin- and (+)-SJ733. The G358S mutation in PfATP4 reduced the affinity of the protein for Na+ and was associated with an increase in the parasite's resting cytosolic Na+ concentration; however, no significant defect in parasite growth rate was observed. Our findings suggest that codon 358 in pfatp4 should be monitored closely in the field as a molecular marker for cipargamin resistance, and that PfATP4 inhibitors in clinical development should be tested for their activity against PfATP4G358S parasites.

Changes in Body Composition and Strength after 12 Weeks of High-Intensity Functional Training with Two Different Loads in Physically Active Men and Women: A Randomized Controlled Study

This study examined the effects of two different resistance loads during high-intensity Functional Training (HIFT) on body composition and maximal strength. Thirty-one healthy young individuals were randomly assigned into three groups: moderate load (ML: 70% 1-RM), low load-(LL: 30% 1-RM), and control (CON). Each experimental group performed HIFT three times per week for 12 weeks with a similar total volume load. Body fat decreased equally in both experimental groups after 6 weeks of training (p < 0.001), but at the end of training it further decreased only in LL compared to ML (−3.19 ± 1.59 vs. −1.64 ± 1.44 kg, p < 0.001), with no change in CON (0.29 ± 1.08 kg, p = 0.998). Lean body mass (LBM) increased after 6 weeks of training (p = 0.019) in ML only, while after 12 weeks a similar increase was observed in LL and ML (1.11 ± 0.65 vs. ML: 1.25 ± 1.59 kg, p = 0.034 and 0.013, respectively), with no change in CON (0.34 ± 0.67 kg, p = 0.991). Maximal strength increased similarly in four out of five exercises for both experimental groups by between 9.5% and 16.9% (p < 0.01) at the end of training, with no change in CON (−0.6 to 4.9%, p > 0.465). In conclusion, twelve weeks of HIFT training with either low or moderate resistance and equal volume load resulted in an equal increase in LBM and maximal strength, but different fat loss.

Resistance status and the rate of resistance development in Musca domestica Linnaeus (Diptera: Muscidae) to permethrin and imidacloprid in Indonesia

A previous report has indicated that in many regions of Indonesia, populations of Musca domestica Linnaeus have shown very high resistance to permethrin and low resistance to imidacloprid. In this study, the resistance status to permethrin and imidacloprid was updated using a topical application and feeding bioassay. Six housefly strains originated from six highly populated cities in Indonesia, namely Serang (SRG), Jakarta (JKT), Bandung (BDG), Semarang (SMG), Yogyakarta (JOG), and Surabaya (SBY). A seventh strain (Danish Pest Infestation Laboratory (DPIL)) served as the control. Each strain was tested for resistance to the two insecticides. In addition, the rate of development of resistance to the two insecticides was measured over ten generations. The results indicated that all field strains showed very high resistance to permethrin. The highest resistance level recorded was in the SRG strain (RR50 = 2880), and the lowest was in the JKT strain (RR50 = 520). Repeated application of permethrin over ten generations increased the resistance level by about 2.7–32.73-fold as compared to the level of their respective parental populations. On the other hand, most strains showed low to moderate resistance to imidacloprid, in which the SRG strain had the highest resistance level (RR50 = 15.5) and the SBY strain had the lowest (RR50 = 2.0). Repeated application of imidacloprid over ten generations increased the resistance level by about 3.25–17.41-fold. The findings, which is the second report of housefly resistance in Indonesia since 2016, provide a crucial foundation for developing appropriate housefly integrated pest management strategies in highly populated areas in Indonesia.

AAC Whitehead hard white spring wheat

AAC Whitehead, an awned hard white spring wheat (Triticum aestivum L.) cultivar, combines high grain yield and good agronomic characteristics with excellent disease resistance. Based on 34 station-years of registration trial data from 2017 to 2019, AAC Whitehead had grain yield significantly higher than all of the check cultivars. AAC Whitehead had maturity similar to the checks, low lodging scores, and significantly shorter plant height than Snowstar and Whitehawk. AAC Whitehead had significantly lower test weight and higher kernel mass than than all the check cultivars. AAC Whitehead expressed resistance to the prevalent races of leaf, stripe and stem rust, and common bunt; and moderate resistance to Fusarium head blight (FHB). It also expressed tolerance to the orange wheat blossom midge. AAC Whitehead expresses quality attributes within the range of the check cultivars and is eligible for grades of Canada Western Hard White Spring wheat.

First Evaluation of Field Evolved Resistance to Commonly Used Insecticides in House Fly Populations from Saudi Arabian Dairy Farms

The house fly, Musca domestica L. (Diptera: Muscidae), is one of the major vectors of several pathogens that affect humans and animals. We evaluated the toxicity of eight insecticides commonly used for house fly control using five field populations collected from dairies in Riyadh, Saudi Arabia. Among the five tested pyrethroids, non to moderate resistance was found in adults of both sexes compared to a susceptible strain. Resistance ratios ranged from 0.5- to 7-fold for alpha-cypermethrin, 2- to 21-fold for deltamethrin, 4- to 19-fold for bifenthrin, 1- to 9-fold for cyfluthrin, and 1- to 8-fold for cypermethrin. Among the three tested organophosphates, low to moderate resistance was found among adult flies compared to the susceptible strain, and the resistance ratios ranged from 4- to 27-fold for fenitrothion, 2- to 14-fold for chlorpyrifos, and 3- to 12-fold for malathion. The median lethal times for the tested insecticides were 3–33 h for alpha-cypermethrin, 3–24 h for deltamethrin, 5–59 h for bifenthrin, 1–7 h for cypermethrin, 0.3–7 h for cyfluthrin, 6–36 h for fenitrothion, 2–21 h for chlorpyrifos, and 3–34 h for malathion. This study presents baseline data pertaining to registered public health insecticides, and the results will assist future studies monitoring insecticide resistance, and the planning of effective integrated vector management programs.

SARS-CoV-2 Delta Variant Displays Moderate Resistance to Neutralizing Antibodies and Spike Protein Properties of Higher Soluble ACE2 Sensitivity, Enhanced Cleavage and Fusogenic Activity

The SARS-CoV-2 B.1.617 lineage variants, Kappa (B.1.617.1) and Delta (B.1.617.2, AY) emerged during the second wave of infections in India, but the Delta variants have become dominant worldwide and continue to evolve. Here, we compared B.1.617 variants for neutralization resistance by convalescent sera, mRNA vaccine-elicited sera, and therapeutic neutralizing antibodies using a pseudovirus neutralization assay. B.1.617.1, B.1.617.2, and AY.1 pseudoviruses showed a modest 1.5- to 4.4-fold reduction in neutralization by convalescent sera and vaccine-elicited sera. In comparison, similar modest reductions were also observed for C.37, P.1, R.1, and B.1.526 pseudoviruses, but 7- and 16-fold reductions for vaccine-elicited and convalescent sera, respectively, were seen for B.1.351 pseudoviruses. Among twenty-three therapeutic antibodies tested, four antibodies showed either complete or partial loss of neutralization against B.1.617.2 pseudoviruses and six antibodies showed either complete or partial loss of neutralization against B.1.617.1 and AY.1 pseudoviruses. Our results indicate that the current mRNA-based vaccines will likely remain effective in protecting against B.1.617 variants. Finally, the P681R substitution confers efficient cleavage of B.1.617 variants’ spike proteins and the spike of Delta variants exhibited greater sensitivity to soluble ACE2 neutralization, as well as fusogenic activity, which may contribute to enhanced spread of Delta variants.

Isolation and molecular identification of microorganisms isolated from soils contaminated with heavy metals in Mosul city

This research is concerned with organisms isolated from soils contaminated with heavy metals in industrial and residential areas in Mosul, the center of Nineveh Governorate, and the diagnosis of these organisms using molecular biology techniques. Samples were collected from four locations in the city between the industrial area and residential neighborhoods. Soil samples were analyzed, and dilutions were prepared, then the dilutions were grown on potato extract and dextrose (PDA) medium for the development of fungi and Nutrient agar for bacterial development. The dilutions were planted by the casting method by three replications, then the process of purifying the fungal and bacterial colonies was carried out using the traditional methods. To diagnose these pure colonies using PCR technique, colonies of fungi were grown on the medium of PDA, and bacteria were grown on the medium of nutritious broth. As a result, nine fungal species were diagnosed; two of them are new undiagnosed genera that have been registered in the gene bank, four of them contain genetic mutations, and three of them are known and previously diagnosed fungi. As for bacteria, two new strains were isolated and registered in the gene bank among the four diagnosed types. And some of these genera exhibited severe resistance to antibiotics, while others showed moderate resistance, in contrast to the control, which was very sensitive to antibiotics.

SARS-COV-2 Delta variant displays moderate resistance to neutralizing antibodies and spike protein properties of higher soluble ACE2 sensitivity, enhanced cleavage and fusogenic activity

The SARS-CoV-2 B.1.617 lineage variants, Kappa (B.1.617.1) and Delta (B.1.617.2, AY) emerged during the second wave of infections in India, but the Delta variants have become dominant worldwide and continue to evolve. The spike proteins of B.1.617.1, B.1.617.2, and AY.1 variants have several substitutions in the receptor binding domain (RBD), including L452R+E484Q, L452R+T478K, and K417N+L452R+T478K, respectively, that could potentially reduce effectiveness of therapeutic antibodies and current vaccines. Here we compared B.1.617 variants, and their single and double RBD substitutions for resistance to neutralization by convalescent sera, mRNA vaccine-elicited sera, and therapeutic neutralizing antibodies using a pseudovirus neutralization assay. Pseudoviruses with the B.1.617.1, B.1.617.2, and AY.1 spike showed a modest 1.5 to 4.4-fold reduction in neutralization titer by convalescent sera and vaccine-elicited sera. In comparison, similar modest reductions were also observed for pseudoviruses with C.37, P.1, R.1, and B.1.526 spikes, but seven- and sixteen-fold reduction for vaccine-elicited and convalescent sera, respectively, was seen for pseudoviruses with the B.1.351 spike. Four of twenty-three therapeutic neutralizing antibodies showed either complete or partial loss of neutralization against B.1.617.2 pseudoviruses due to the L452R substitution, whereas six of twenty-three therapeutic neutralizing antibodies showed either complete or partial loss of neutralization against B.1.617.1 pseudoviruses due to either the E484Q or L452R substitution. Against AY.1 pseudoviruses, the L452R and K417N substitutions accounted for the loss of neutralization by four antibodies and one antibody, respectively, whereas one antibody lost potency that could not be fully accounted for by a single RBD substitution. The modest resistance of B.1.617 variants to vaccine-elicited sera suggest that current mRNA-based vaccines will likely remain effective in protecting against B.1.617 variants, but the therapeutic antibodies need to be carefully selected based on their resistance profiles. Finally, the spike proteins of B.1.617 variants are more efficiently cleaved due to the P681R substitution, and the spike of Delta variants exhibited greater sensitivity to soluble ACE2 neutralization, as well as fusogenic activity, which may contribute to enhanced spread of Delta variants.

Nitrogenous Fertilizer Reduces Resistance but Enhances Tolerance to the Brown Planthopper in Fast-Growing, Moderately Resistant Rice

The brown planthopper, Nilaparvata lugens (Stål), is a key challenge to rice production in Asia. Outbreaks of planthoppers are associated with excessive fertilizer applications; consequently, we examined planthopper interactions with susceptible, tolerant and resistant varieties of rice under varying levels of soil nitrogen in a greenhouse experiment. We compared planthopper fitness (survival × reproduction) and plant tolerance (functional plant loss index) for 16 varieties at 0, 80 and 150 Kg added nitrogen ha−1. The planthoppers grew larger, developed more quickly and laid more eggs on susceptible varieties, compared with the resistant and tolerant varieties. Moreover, soil nitrogen generally increased planthopper fitness on resistant varieties, but relative resistance was maintained. Functional plant loss was highest among the susceptible varieties, but weight and growth rate reductions per mg of planthopper were often highest in the tolerant varieties. Tolerance was associated with large, fast-growing plants, with at least moderate resistance to the planthopper. Susceptibility was associated with a small size and/or an absence of resistance genes. Our results suggested that early-tillering rice plants can be both resistant and tolerant to the brown planthopper, but cannot be both susceptible and tolerant of planthoppers at high densities. This indicates that at least moderate resistance is required for tolerance against this herbivore. Furthermore, although dwarf varieties had a low tolerance of planthoppers, they could express resistance through functioning resistance genes.

Aromatic Rice with Multiple Disease Resistance Developed through Anther Culture

The experiments were conducted during kharif (June to September) 2018 and 2019 to screen 7 aromatic rice double haploid (DH) lines developed against Bacterial Leaf Bligh (BLB) and blast disease. Three genes Xa21, xa13, xa5 for BLB resistance and two genes Pi1, Pi2 for blast resistance was introgressed into a local aromatic rice variety Dubraj from Chhattisgarh State of India. Dubraj is extremely popular aromatic short medium grain variety in Chhattisgarh and Madhya Pradesh region and hence used in this study. RPBIO-226 and PR122 rice variety is used as donor for BLB and blast resistance respectively. 7 DH lines (L-1 to L-7) were developed from the crosses (Dubraj X RPBIO-226 X PR122) in the year 2017. Phenotypic evaluation for BLB diseases resistance has shown that L-1, L-6 and L-7 (score 1) has a similar resistance as the control check RPBIO-226(score-1) and 3 lines with moderate resistance (L-3 score 2) (L-4 score 3) (L-5 score 3). For blast disease, three lines (L-4, L-6 and L-7) were identified as resistance with the disease score of 1, whereas three were grouped under the section moderately resistant (L-1, L-3 and L- 5) with a disease score of 3. Sensory test (1.7% KOH) was conducted for the estimation of presence / absence of aroma. Moderately strong aroma was present in four lines (L-1, L-3, L-6 and L-7). On the basis of yield and presence of aroma in grain, L-3 has been selected with moderately resistant to BLB and blast for further advancement and subsequently evaluated under State Initial Evaluation Trial (SIET) during kharif 2020. Within 3 years of development of DH line, L-3 is under varietal developmental trail, which itself indicate the efficacy of anther culture in fixing homozygosity and speedy development of desired variety within short period of time.