Impact of Exposure to Leaves From Metal-Polluted Sites on the Developmental Parameters of Larvae of the Dark Sword-Grass, Agrotis ipsilon (Lepidoptera: Noctuidae).

Insects are impacted by pollutants in their environments and food sources. Herein, we set out a semi-field study to assess the impact of environmental heavy metal contamination on developmental parameters, energy reserves, and acidic and alkaline phosphatases in the larval Agrotis ipsilon (Lepidoptera: Noctuidae). Castor leaves from contaminated and uncontaminated (reference site) areas were fed to A. ipsilon larvae in all treatments. The heavy metal concentrations in the plant from different areas (contaminated and reference sites) and in the larvae were analyzed. Toxic effects were observed in the larvae feeding on the leaves from the metal contaminated areas. Larval and pupal weights, growth indices, and larval fitness were all significantly lower than in the reference group. Likewise, in the third and fourth instars, there was a significant decrease in both the survival and moth emergence rates. In contrast, the pupation duration was significantly longer. Total protein, lipid, and glycogen content showed significant reductions in treated larvae. Larval homogenate samples contaminated with heavy metals showed a significant increase in acid- and alkaline- phosphatase levels. The results obtained could provide a basis for a long-term evaluation of the risk associated with heavy metals and their impact on plant populations and important agricultural pests.

Phosphodiesterase-5 inhibitors use and the risk of alzheimer's disease: a systematic review and meta-analysis.

BACKGROUND: The management of Alzheimer's disease (AD) poses considerable challenges, necessitating the pursuit of innovative therapeutic approaches. Recent research has spotlighted the promising role of phosphodiesterase type 5 inhibitors (PDE5Is) in reducing the prevalence of AD, utilizing their vasodilatory properties to suggest a potential neuroprotective effect. This meta-analysis and systematic review aims to assess the relationship between the use of PDE5Is and the risk of AD. METHODS: A detailed examination was carried out across several electronic databases till March 2024, including PubMed, Web of Science, Scopus, CENTRAL, and Embase. The focus was on identifying studies that compare the occurrence of AD among PDE5I users vs non-users. Through a random-effects model, pooled hazard ratios (HRs) were calculated, in alignment with guidelines from the Cochrane Handbook for Systematic Reviews and Meta-Analysis and the PRISMA standards. RESULTS: This analysis included six studies, cumulating a participant count of 8,337,313, involving individuals treated with sildenafil, tadalafil, and vardenafil, against a control group undergoing other or no treatments. The cumulative HR for AD risk among PDE5I users versus the control group was 0.53 (95% CI: 0.32-0.86, p = 0.008), signaling a markedly reduced likelihood of AD development in the PDE5I group. Particularly, sildenafil usage showed a significant risk reduction (HR: 0.46, 95% CI: 0.31-0.70, p < 0.001), while findings for tadalafil and vardenafil were not significant. Test of subgroup differences found no difference between male and female participants in the risk of AD. CONCLUSIONS: Our findings suggest that the use of PDE5Is is associated with a reduced risk of AD, highlighting its potential as a protective agent against neurodegenerative diseases. Given the very low quality of evidence and the heterogeneity among the included studies, further high-quality research is warranted to confirm these findings and elucidate the underlying mechanisms. Register number PROSPERO 2024: CRD42024522197.

Development of nanofibrous scaffolds containing polylactic acid modified with turmeric and hydroxyapatite/vivianite nanoparticles for wound dressing applications.

Damaging the outer layer of the body (the skin) has been a common issue for decades. Fabrication of nanofibrous membranes via the electrospinning technique for the sake of making the wound healing process more facile has caught a lot of interest. For this purpose, a polymeric scaffold of polylactic acid (PLA) was doped with nanoparticles with different concentrations of turmeric/hydroxyapatite/vivianite/graphene oxide. The obtained membrane was tested by XRD, SEM, FTIR, and XPS. The surface topography of the scaffold has experienced changes upon adding different concentrations of the nanoparticles. The contact angle was measured by water droplets. It accentuated change in CA starting from 43.9o for pure condition of PLA to 67.7o for PLA/turmeric/vivianite. The thermogravimetric analysis (TGA) test stated that the PLA scaffold features are thermally stable in relatively high-temperature conditions initiating from room temperature to about 300 °C, meeting the maximum loss in mass of about 5 %. The cell viability was carried out in prepared vitro for the sample which contains PLA/turmeric/vivianite/GO, it was elucidated that the IC50 was around 3060 µg/ml.

Stage-, sex- and tissue-related changes in H2O2, glutathione concentration, and glutathione-dependent enzymes activity in Aiolopus thalassinus (Orthoptera: Acrididae) from heavy metal polluted areas.

This study is part of a large project carried out at the Cairo University, Egypt, and focused on assessing physiological and biochemical changes in Aiolopus thalassinus under the influence of environmental pollution with heavy metals (Pb, Cd, Cu, and Zn). The study aimed to investigate parameters related to maintaining redox balance, with particular emphasis on stage-, sex- and tissue-dependent differences in H2O2 and glutathione (GSH) levels and activity of selected enzymes involved in GSH metabolism. A noticeable increase in the concentration of H2O2 was found, especially in the gut of 5th instar nymphs and females from the highly polluted site. An increase in GSH concentration was significant, especially in the gut of adult A. thalassinus from the high polluted site. However, recycling of reduced form of glutathione in the gut by glutathione reductase (GR) was relevant only for females from the high polluted site. Nymphs and females generally showed higher glutathione S-transferase (GST) activity, especially in the gut. These stage- and sex-related differences can result from different growth dynamic and various reproductive functions of nymphs and both sexes. The digestive track is in direct contact with xenobiotics consumed with food. Nymphs are characterized by vigorous growth, they feed intensively, and their development processes are associated with substantial oxygen consumption. Also, maintaining the antioxidant system at a high level can be more important for females than males due to egg production over a long period. It appears that de novo GSH synthesis is a favorable and cost-effective adaptation mechanism for A. thalassinus living in the high polluted site.

Antioxidant enzyme activity in responses to environmentally induced oxidative stress in the 5th instar nymphs of Aiolopus thalassinus (Orthoptera: Acrididae).

The response of antioxidant enzymes to oxidative environmental stress was determined in 5th instar nymphs of Aiolopus thalassinus (Orthoptera: Acrididae) collected from sites with different level of pollution with heavy metals, PO43-, and SO42-. The high polluted site induced higher DNA damage to individuals compared to the control site. The highest values of tail length (TL), tail moment (TM), and percent of DNA in tail (TDNA) were found in the gut of 5th instar nymphs from a high polluted site. Also, protein carbonyls and lipid peroxide levels were significantly higher in insects collected from polluted sites compared to those from the control site. A strong positive correlation between both protein carbonyl and lipid peroxide concentration and the pollution level of the sites was found in all tissues of the insects. The activity of superoxide dismutase (SOD) in the brain of insects collected from the high polluted site was significantly higher than that in the thoracic muscles and gut. We observed strong inhibition of catalase (CAT) activity. This effect was apparently caused by pollutants present at the high polluted site. The level of pollution significantly influenced polyphenol oxidase (PPO) activity in A. thalassinus nymphs in all examined tissues. The highest values were observed in the brain. The relationship between pollution and ascorbate peroxidase (APOX) activity in the examined tissues had no clear tendency. However, the lowest APOX activity was observed in individuals from the low polluted site. Level of pollution of sampling sites, oxidative stress biomarkers, and enzymatic response in A. thalanthsis 5th instar were negatively or positively correlated. Oxidative damage parameters, especially the percent of severed cells, lipid peroxides, and the activity of APOX, can be perceived as good markers of environmental multistress.

The multilevel antibiotic-induced perturbations to biological systems: Early-life exposure induces long-lasting damages to muscle structure and mitochondrial metabolism in flies.

Antibiotics have been increasingly used over the past decades for human medicine, food-animal agriculture, aquaculture, and plant production. A significant part of the active molecules of antibiotics can be released into the environment, in turn affecting ecosystem functioning and biogeochemical processes. At lower organizational scales, these substances affect bacterial symbionts of insects, with negative consequences on growth and development of juveniles, and population dynamics. Yet, the multiple alterations of cellular physiology and metabolic processes have remained insufficiently explored in insects. We evaluated the effects of five antibiotics with different mode of action, i.e. ampicillin, cefradine, chloramphenicol, cycloheximide, and tetracycline, on the survival and ultrastructural organization of the flight muscles of newly emerged blow flies Chrysomya albiceps. Then, we examined the effects of different concentrations of antibiotics on mitochondrial protein content, efficiency of oxidative phosphorylation, and activity of transaminases (Glutamate oxaloacetate transaminase and glutamate pyruvate transaminase) and described the cellular metabolic perturbations of flies treated with antibiotics. All antibiotics affected the survival of the insects and decreased the total mitochondrial protein content in a dose-dependent manner. Ultrastructural organization of flight muscles in treated flies differs dramatically compared to the control groups and severe pathological damages/structures disorganization of mitochondria appeared. The activities of mitochondrial transaminases significantly increased with increased antibiotic concentrations. The oxidation rate of pyruvate + proline from isolated mitochondria of the flight muscles of 1-day-old flies was significantly reduced at high doses of antibiotics. In parallel, the level of several metabolites, including TCA cycle intermediates, was reduced in antibiotics-treated flies. Overall, antibiotics provoked a system-wide alteration of the structure and physiology of flight muscles of the blow fly Ch. albiceps, and may have fitness consequences at the organism level. Environmental antibiotic pollution is likely to have unwanted cascading ecological effects of insect population dynamics and community structure.

Biomonitoring of genotoxicity of industrial fertilizer pollutants in Aiolopus thalassinus (Orthoptera: Acrididae) using alkaline comet assay.

Phosphate fertilizer industry is considered as one of the main sources of environmental pollutants. Besides solid waste products, e.g. phosphates, sulphates, and heavy metals, also atmospheric pollutants, such as hydrofluoric acid fumes (HF), sulphur dioxide (SO2), nitrogen oxides (NO2), and particulate matter with diameter up to 10 µm (PM10) can be dangerous. Genotoxic effect of these pollutants was monitored by assessing the DNA damage using alkaline comet assay on cells from brain, thoracic muscles and gut of Aiolopus thalassinus collected at three sites (A-C) located at 1, 3, and 6 km away from Abu-Zaabal Company for Fertilizers and Chemical Industries. Control site was established 32 km from the source of pollution, at the Cairo University Campus. The level of the DNA damage was significantly higher in insects from polluted sites comparing to that from the control site. A strong negative correlation between percentage of cells with visible DNA damage (% of severed cells) and the distance of the sites from Abu-Zaabal Company was found. The best parameter for monitoring of fertilizer pollutants is % of severed cells. Possible impact of Abu-Zaabal Company (extremely high concentration of phosphates and sulphates in all the polluted sites) on DNA integrity in A. thalassinus tissues was discussed. The potential use of the comet assay as a biomonitoring method of the environmental pollution caused by fertilizer industry was proposed. Specific pollution resulting from the activity of the fertilizer industry can cause comparable adverse effects in the organisms inhabiting areas up to 6 km from the source of contamination.

Evaluation of oxidative stress biomarkers in Aiolopus thalassinus (Orthoptera: Acrididae) collected from areas polluted by the fertilizer industry.

The waste products of the fertilizer industry such as heavy metals, but especially phosphates and sulphates, are a serious problem that influences the structure and functioning of ecosystems. The levels of Cd, Pb, Zn, Cu, sulphates and phosphates were measured in soil samples from four sites: a control and sites that were 1, 3 and 6 km (sites A-C) away from the Abu-Zaabal Fertilizer Company. Oxidative stress markers (protein carbonyls, lipid peroxides), antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), polyphenoloxidase (PPO) and ascorbate peroxidase (APOX)) were evaluated in the tissues of Aiolopus thalassinus, collected from the corresponding sites. The highest concentrations of Cu and Zn were found in the soil from site A. The level of protein carbonyls in the brain, thoracic muscles and gut of the males and females from sites A, B and C were 11.82, 4.38, 5.97 (males) and 19.04, 16.65, 7.79 (females) times higher, respectively, compared to the individuals from the control site. Lipid peroxides levels in both sexes were significantly correlated with the distance from the source of the contamination. In the brain, thoracic muscles and gut of the males and females collected from site A, the level of lipid peroxides were 15.41, 23.49, 11.50 (males) and 25.36, 11.34, 15.37 (females) times higher compared to the values of the control animals. The activities of SOD, PPO, CAT and APOX were significantly affected by the environmental pollutants. The enzymatic and non-enzymatic oxidative markers in the Aiolopus thalassinus, a common insect species that inhabits various ecosystems, can also be used as a relevant biomarker of the pollution that is caused by the fertilizer industry.

Molecular characterization of a c-type lysozyme from the desert locust, Schistocerca gregaria (Orthoptera: Acrididae).

Lysozymes are bacteriolytic peptides that are implicated in the insect nonspecific innate immune responses. In this study, a full-length cDNA encoding a c-type lysozyme from Schistocerca gregaria (SgLys) has been cloned and characterized from the fat body of immune-challenged 5(th) instar. The deduced mature lysozyme is 119 amino acid residues in length, has a calculated molecular mass of 13.4 kDa and an isoelectric point (Ip) of 9.2. SgLys showed high identities with other insect lysozymes, ranging from 41.5% to 93.3% by BLASTp search in NCBI. Eukaryotic in vitro expression of the SgLys ORF (rSgLys) with an apparent molecular mass of ∼16 kDa under SDS-PAGE is close to the calculated molecular weight of the full-length protein. rSgLys displayed growth inhibitory activity against Gram-negative and Gram-positive bacteria. 3D structure modeling of SgLys, based on comparison with that of silkworm lysozyme, and sequence comparison with the helix-loop-helix (α-hairpin) structure of hen egg white lysozyme (HEWL) were employed to interpret the antibacterial potencies. Phylogenetic alignments indicate that SgLys aligns well with insect c-type lysozymes that expressed principally in fat body and hemocytes and whose role has been defined as immune-related. Western blot analysis showed that SgLys expression was highest at 6-12 h post-bacterial challenge and subsequently decreased with time. Transcriptional profiles of SgLys were determined by semi-quantitative RT-PCR analysis. SgLys transcript was upregulated at the highest level in fat body, hemocytes, salivary gland, thoracic muscles, and epidermal tissue. It was expressed in all developmental stages from egg to adult. These data indicate that SgLys is a predominant acute-phase protein that is expressed and upregulated upon immune challenge.

Profiling of proteins and proteases in the products of the salivary gland, digestive tract and excretions from larvae of the camel nasal botfly, Cephalopina titillator (Clark).

Proteins and proteolytic activities in the contents of the salivary gland (SGc), digestive tract (DTc) and excretory-secretory products (ESP) from larvae of the camel nasal botfly Cephalopina titillator were separated electrophoretically, and characterized. The protein profiles of the different samples were qualitatively quite similar in the larval stages L2 and L3. Zymogram analysis of proteases in the samples indicated that the digestive tract contained a greater variety of proteases than the salivary gland or the excretory-secretory products. They are mainly serine proteases. Proteases of ESP and DTc (especially of 3rd instar) contain trypsin- and chymotrypsin-like serine proteases, while the serine proteases of SGc are not of the trypsin- or chemotrypsin-type.

Isolation, characterization, kinetics, and enzymatic and nonenzymatic microbicidal activities of a novel c-type lysozyme from plasma of Schistocerca gregaria (Orthoptera: Acrididae).

A protein, designated as Sgl, showing a muramidase lytic activity to the cell wall of the Gram-positive bacterium Micrococcus lysodeikticus was isolated for the first time from plasma of Escherichia coli-immunized fifth instar Schistocerca gregaria. The isolated Sgl was detected as a single protein band, on both native- and SDS-PAGE, has a molecular weight of ∼15.7 kDa and an isoelectric point (pI) of ca 9.3 and its antiserum has specifically recognized its isolated form. Fifty-nine percentage of Sgl lytic activity was recovered in the isolated fractions and yielded ca 126-fold increase in specific activity than that of the crude. The partial N-terminal amino acid sequence of the Sgl has 55 and 40% maximum identity with Bombyx mori and Gallus gallus c-type lysozymes, respectively. The antibacterial activity against the Gram-positive and the Gram-negative bacteria were comparatively stronger than that of the hen egg white lysozyme (HEWL). The detected Sgl poration to the inner membrane that reach a maximum ability after 3 h was suggested to operate as a nonenzymatic mechanism for Gram-negative bacterial cell lysis, as tested in a permease-deficient E. coli, ML-35 strain. Sgl showed a maximal muramidase activity at pH 6.2, 30-50°C, and 0.05 M Ca(2+) or Mg(2+); and has a Km of 0.5 µg/ml and a Vmax of 0.518 with M. lysodeikticus as a substrate. The Sgl displayed a chitinase activity against chitin with a Km of 0.93 mg/ml and a Vmax of 1.63.