The Impact of Graphite Oxide Nanocomposites on the Antibacterial Activity of Serum.

Nanoparticles can interact with the complement system and modulate the inflammatory response. The effect of these interactions on the complement activity strongly depends on physicochemical properties of nanoparticles. The interactions of silver nanoparticles with serum proteins (particularly with the complement system components) have the potential to significantly affect the antibacterial activity of serum, with serious implications for human health. The aim of the study was to assess the influence of graphite oxide (GO) nanocomposites (GO, GO-PcZr(Lys)2-Ag, GO-Ag, GO-PcZr(Lys)2) on the antibacterial activity of normal human serum (NHS), serum activity against bacteria isolated from alveoli treated with nanocomposites, and nanocomposite sensitivity of bacteria exposed to serum in vitro (using normal human serum). Additionally, the in vivo cytotoxic effect of the GO compounds was determined with application of a Galleria mellonella larvae model. GO-PcZr(Lys)2, without IR irradiation enhance the antimicrobial efficacy of the human serum. IR irradiation enhances bactericidal activity of serum in the case of the GO-PcZr(Lys)2-Ag sample. Bacteria exposed to nanocomposites become more sensitive to the action of serum. Bacteria exposed to serum become more sensitive to the GO-Ag sample. None of the tested GO nanocomposites displayed a cytotoxicity towards larvae.

Impact of artificial light intensity on nocturnal insect diversity in urban and rural areas of the Asir province, Saudi Arabia.

Continuous urban developments have resulted in increased demand for street furniture, one of which is street light columns. Artificial light at night (ALAN) pose significant impacts on insect diversity in urban and rural areas. The ALAN is a significant driver of decline in insect diversity. This study evaluated the impact of light intensity and sky quality at night on insect diversity in rural and urban areas of the Asir province, Saudi Arabia. Insect traps were installed in both areas during night. Light intensity of nearby road lamps was measured using light meter, while sky quality was measured using sky quality meter. Rural areas exhibited low light intensity (10.33 flux/f.candle) and good sky quality (18.80 magnitude/arcsec2). Urban areas exhibited intense light (89.33 flux/f.candle) and poor sky quality (15.49 magnitude/arcsec2). Higher insect diversity was recorded for rural areas where insects belonging to seven orders (i.e., Diptera, Lepidoptera, Hemiptera, Hymenoptera, Coleoptera, Neuroptera, and Dermaptera) were collected. However, insects of four orders (i.e., Diptera, Lepidoptera, Hemiptera, and Neuroptera) were found in urban areas indicating low diversity. Lepidopteran insects were frequently recorded from rural areas indicating they are attracted to artificial light. It is concluded that excessive ALAN and poor sky quality at night disrupt insect biodiversity. Therefore, ALAN and sky quality must be considered responsible for decline in insect biodiversity along with other known factors.

Effects of 200 Gy 60Co-γ Radiation on the Regulation of Antioxidant Enzymes, Hsp70 Genes, and Serum Molecules of Plutella xylostella (Linnaeus).

The diamondback moth, Plutella xylostella (Linnaeus), is one of the notorious pests causing substantial loses to many cruciferous vegetables across the nations. The effects of 60Co-γ radiation on physiology of P. xylostella were investigated and the results displayed that 200 Gy irradiation significantly alters the antioxidant enzyme regulation in six-day-old male pupae of P. xylostella. First, in our research, we detected Oxidase system and stress response mechanism of irradiated pupae, the results displayed that 200 Gy irradiation significantly alters the antioxidant enzyme regulation in six-day-old male pupae of P. xylostella. The levels of superoxide dismutase (SOD) and catalase (CAT) were increased significantly in contrast the level of peroxidase (POD) and glutathione S-transferase (GST) were decreased in 12⁻24 h post-treatment. The heat shock proteins (Hsps) gene expression level was significant increasing, maximum > 2-folds upregulation of genes were observed in peak. However, they also had a trend of gradual recovery with development. Second, we detected the testis lactate dehydrogenase (LDH) and acid phosphatase (ACP) activity found that in male adults testis they increased significantly than control during its development. Thus the present research investigation highlights that the 60Co-γ radiation treatments alters the physiological development of diamondback moth. The results showed that 200 Gy dosage resulted in stress damage to the body and reproductive system of the diamondback moth.

Intrinsic attenuation of post-irradiation calcium and ER stress imparts significant radioprotection to lepidopteran insect cells.

Sf9 lepidopteran insect cells are 100-200 times more radioresistant than mammalian cells. This distinctive feature thus makes them suitable for studies exploring radioprotective molecular mechanisms. It has been established from previous studies of our group that downstream mitochondrial apoptotic signaling pathways in Sf9 cells are quite similar to mammalian cells, implicating the upstream signaling pathways in their extensive radioresistance. In the present study, intracellular and mitochondrial calcium levels remained unaltered in Sf9 cells in response to radiation, in sharp contrast to human (HEK293T) cells. The isolated mitochondria from Sf9 cells exhibited nearly 1.5 times greater calcium retention capacity than mammalian cells, highlighting their inherent stress resilience. Importantly, UPR/ER stress marker proteins (p-eIF2α, GRP4 and SERCA) remained unaltered by radiation and suggested highly attenuated ER and calcium stress. Lack of SERCA induction further corroborates the lack of radiation-induced calcium mobilization in these cells. The expression of CaMKII, an important effector molecule of calcium signaling, did not alter in response to radiation. Inhibiting CaMKII by KN-93 or suppressing CaM by siRNA failed to alter Sf9 cells response to radiation and suggests CaM-CaMKII independent radiation signaling. Therefore, this study suggests that attenuated calcium signaling/ER stress is an important determinant of lepidopteran cell radioresistance.

Evidence for a radiation-responsive 'p53 gateway' contributing significantly to the radioresistance of lepidopteran insect cells.

Recently, we have demonstrated that microRNA-31 (miR-31) overexpression is inherent to radiation-induced cell death in the highly radioresistant Sf9 insect cells, and regulates pro-apoptotic Bax translocation to mitochondria. In the present study, we report that at sub-lethal radiation doses for Sf9 cells, miR-31 is significantly downregulated and is tightly regulated by an unusual mechanism involving p53. While ectopic overexpression of a well-conserved Sfp53 caused typical apoptosis, radiation-induced p53 accumulation observed selectively at sub-lethal doses failed to induce cell death. Further investigation of this paradoxical response revealed an intriguing phenomenon that sub-lethal radiation doses result in accumulation of a 'hyper-phosphorylated' Sfp53, which in turn binds to miR-31 genomic location and suppresses its expression to prevent cell death. Interestingly, priming cells with sub-lethal doses even prevented the apoptosis induced by lethal radiation or ectopic Sfp53 overexpression. On the other hand, silencing p53 increased radiation-induced cell death by inhibiting miR-31 downregulation. This study thus shows the existence of a unique radiation-responsive 'p53 gateway' preventing miR-31-mediated apoptosis in Sf9 cells. Since Sfp53 has a good functional homology with human p53, this study may have significant implications for effectively modulating the mammalian cell radioresistance.

Alterations of Immune Parameters on Trichoplusia ni (Lepidoptera: Noctuidae) Larvae Exposed to Extremely Low-Frequency Electromagnetic Fields.

Worldwide mobile telephone and microwave use have resulted in an increasing presence of extremely low-frequency electromagnetic field radiations (ELF-EMFs) in ecosystems. ELF-EMFs have been associated with altered physiological processes that can adversely affect exposed organisms. In this study, Trichoplusia ni Hübner larvae were exposed for 24, 48, or 72 h to ELF-EMFs (60 Hz and 2.0 mT) to assess effects on immune response parameters and fertility. Trichoplusia ni life cycle and fertility were not affected by 24-h exposure. However, the number of apoptotic-like cells and cellular immune response significantly increased (P < 0.01) after 72-h exposure (2- and 1.1-fold, respectively), whereas hemolymph total protein and hemocyte cells were reduced (P < 0.01; 16 and 50%, respectively) after 48-h exposure. Hemocyte cell type analysis resulted in significantly (P < 0.01) higher granulocytes number in the unexposed (2-fold increase) and oenocytoids in the 72-h-exposed larvae (28.6-fold increase). Quantitative retrotranscription (RT-qPCR) showed that after 72-h ELF-EMF exposure, the antimicrobial peptides cecropin, lysozyme, gallerimycin, and pgrp were downregulated by 24,866.0, 2.69-, 119.1-, and 1.45-fold, respectively, whereas attacin and defensin were upregulated by 1.59- and 1.85-fold, respectively. The effect of ELF-EMFs on the T. ni larvae immune response and their potential impact on its physiology and susceptibility to pathogens are discussed. This information may provide new insight of ELF-EMFs on other pest species, as well as for the preservation of ecologically important species.

Effect of gamma radiation on Phenoloxidase pathway, antioxidant defense mechanism in Helicoverpa armigera (Lepidoptera: Noctuidae) and its implication in inherited sterility towards pest suppression.

PURPOSE: To investigate age-correlated radiosensitivity in highly radioresistant lepidopteran pest, Helicoverpa armigera, upon exposure to ionizing radiation and to examine the irradiation impact on stress-molecular responses in F1 (first-filial) progeny of irradiated (100 Gy) male moths in relation to its reproductive behavior. MATERIALS AND METHODS: Efficacy of sub-lethal gamma radiation was evaluated on two markedly apart ontogenic stages, neonates and adult moths. Differential growth, reproductive behavior and stress-indicating molecular responses were examined upto F1 progeny of sub-sterilized moths. Free-radical scavenging enzymes, superoxide dismutase (SOD), catalase (CAT) and Phenoloxidase cascade enzymes, pro-phenoloxidase (PPO), its activating enzyme (PPAE) were studied in irradiated and irradiated plus microbial challenge regimen (dual-stress) by Real-time RT-PCR (reverse-transcription-polymerase-chain-reaction). RESULTS: An inverse correlation of radiosensitivity with developmental age of insect was observed. F1 sterility was higher than parent sterility. F1 progeny exhibited protraction in development and decreased survival upon irradiation. Sex ratio in F1 progeny was skewed towards males. PPO, PPAE, SOD and CAT transcripts were downregulated upon neonate irradiation resulting in enhanced vulnerability of larvae to incidental microbial challenge. These transcripts were upregulated in F1 progeny of sub-sterilized male moths (100 Gy) upon dual-stress. CONCLUSIONS: Irradiation impact on stress-indicating molecular responses in F1 progeny is correlated with its reproductive performance. These observations will permit defining regimen having pragmatic viability of 'F1 sterility technique' for pest suppression. Gamma dose of 100 Gy would ensure balance between induced sterility of males and their field competitiveness. These parameters would facilitate integration of biocontrol strategy with parabiological 'Sterile Insect Release Technique'.

Bioenergetics growth model for the effect of gamma irradiation and plant extract (Barnoof) on the progeny of Black cutworm, Agrotis ipsilon (Hufnagel).

This study investigated the effects of two substerilizing doses of gamma radiation, 100 and 150 Gray (Gy), and/or the plant extract Conyza dioscorides (Barnoof) in two solvents on certain biological aspects and the energy budget of the Black cutworm, Agrotis ipsilon (Hufnagel). Data revealed that the treatment combination of gamma radiation with the plant extract significantly increased reproduction compared with the control at all treatment levels (doses and concentrations). Most of the treatments increased the values of developmental/day and larval duration in the F(1) progeny compared with the progeny from the control treatment, although the percentage survival was decreased in all treatments. These values were more obvious in the combination treatments compared with either gamma radiation or plant extract treatments alone. The coefficient of metabolizable energy (C.M.E.) was not affected by any treatments. No consistent effect on the efficiency of storage of ingested energy [E.S.I.(E.)] and the efficiency of storage of metabolizable energy [E.S.M.(E.)] when the F(1) progeny were treated with plant extract alone or with the plant extract combined with the 100 Gy dose of radiation was noted, while they were both significantly increased at all treatment levels when the 150 Gy dose of radiation was combined with the plant extract. The results obtained are discussed in terms of their implications for the best substerilizing dose of radiation on parental male pupae of A. ipsilon.

A calcium-insensitive attenuated nitrosative stress response contributes significantly in the radioresistance of Sf9 insect cells.

Lepidopteran insects/insect cells display 50-100 times higher radioresistance than humans, and are evolutionarily closest to mammals amongst all radioresistant organisms known. Compared to mammalian cells, Lepidopteran cells (TN-368, Sf9) display more efficient antioxidant system and DNA repair and suffer considerably less radiation-induced DNA/cytogenetic damage and apoptosis. Recent studies indicate that a considerably lower radiation-induced oxidative stress may significantly reduce macromolecular damage in Lepidopteran cells. Since nitrosative stress contributes in radiation-induced cellular damage, we investigated its nature in the γ-irradiated Sf9 cells (derived from Spodoptera frugiperda; order Lepidoptera; family Noctuidae) and compared with BMG-1 human cell line having significant NOS expression. Radiation induced considerably less ROS/RNS in Sf9 cells, which remained unchanged on treatment with NOS inhibitor l-NMMA. Surprisingly, growth of Sf9 cultures or irradiation could not induce NO or its metabolites, indicating negligible basal/radiation-induced NOS activity that remained unchanged even after supplementation with arginine. Cytosolic calcium release following high-dose (1000-2000Gy at 61.1cGys(-1)) γ-irradiation or H(2)O(2) (250µM) treatment also failed to generate NO in Sf9 cells having high constitutive levels of calmodulin, whereas BMG-1 cells displayed considerable calcium-dependent NO generation even following 10Gy dose. These results strongly imply the lack of calcium-mediated NOS activity in Sf9 cells. Addition of exogenous NO from GSH-NO caused considerable increase in radiation-induced apoptosis, indicating significant contribution of constitutively attenuated nitrosative stress response into the radioresistance of Lepidopteran cells. Our study demonstrates for the first time that a calcium-insensitive, attenuated nitrosative stress response may contribute significantly in the unusual radioresistance displayed by Lepidopteran insect cells.

[Influence of ionizing radiation on the formation of adult antennae in large fruit-tree tortrix Archips podana Scop. (Lepidoptera, Tortricidae)].

The influence of sterilizing doses of X-radiation on the formation of adult antennae during the pupa period of development of Archips podana was studied. We found a change in the number of coeloconic and auriculate sensilla in those imago whose pupae were exposed to radiation. A slow-down in the process of antennae formation at the pupal stage was registered after the irradiation of instar five larvae and newly formed pupae.

[Effect of ionizing radiation on oocyte development and reproductive activity in Archips Podana scop. (Lepidoptera, Tortricidae)].

The influence of ionizing radiation on oocyte development and male reproductive success in Archips podana has been studied. The increase of the percentage of the vitellogenous oocyte and the decrease of the percentage of the chorion oocyte against the control have been shown. Additionally, increases in the percentage of sterile females and the number of sterile eggs have been pointed out when control females mated with irradiated males.

Effect of irradiation on Mexican leafroller (Lepidoptera: Tortricidae) development and reproduction.

The effects of irradiation on egg, larval, and pupal development, and adult reproduction in Mexican leafroller, Amorbia emigratella Busck (Lepidoptera: Tortricidae), were examined. Eggs, neonates, early instars, late instars, early pupae, and late pupae were irradiated at target doses of 60, 90, 120, or 150 Gy, or they were left untreated as controls in replicated factorial experiments. Survival to the adult stage was recorded. Tolerance to radiation increased with increasing age and developmental stage. A radiation dose of 90 Gy applied to neonates and early instars prevented adult emergence. A dose of 150 Gy was not sufficient to prevent adult emergence in late instars or pupae. The effect of irradiation on sterility was examined in late pupae and adult moths. For progeny produced by insects treated as late pupae, a total of three out of 3,130 eggs hatched at 90 Gy, 0 out of 2,900 eggs hatched at 120 Gy, and 0 out of 1,700 eggs hatched at 150 Gy. From regression analysis, the dose predicted to prevent egg hatch from the progeny of irradiated late pupae was 120 Gy, with a 95% confidence interval of 101-149 Gy. The late pupa is the most radiotolerant stage likely to occur with exported commodities; therefore, a minimum absorbed radiation dose of 149 Gy (nominally 150 Gy) has potential as a quarantine treatment. Reciprocal crosses between irradiated and unirradiated moths demonstrated that males were more radiotolerant than females. Irradiation of female moths at a target dose of 90 Gy before pairing and mating with irradiated or unirradiated males resulted in no viable eggs, whereas irradiated males paired with unirradiated females produced viable eggs at 90 and 150 Gy.

Eyes of male and female Orgyia antiqua (Lepidoptera; Lymantriidae) react differently to an exposure with UV-A.

The structural organization of the eyes belonging to 12 winged male and 12 wingless female Orgyia antiqua moths, exposed for 1 h to UV-radiation (lambda(max)=351 nm) of 1.4 kW/m2, was compared with that of 12 male and 12 female non-irradiated control specimens. Following the UV-exposure, the screening pigments were found in a position indicative of extreme light-adaptation. Extensive formations of vesicles along the perimeter of the cones as well as disintegrating ER in the cone cytoplasm were noticeable, especially in the eye of the female. On the retinal side of the clearzone, the microvilli of the rhabdoms had become affected by the UV in characteristic ways: in the male eye, retinal cell damage in the form of microvillar swellings and disintegrations were largely confined to just two cells per ommatidium, placed opposite to each other. The female eye, once again, exhibited greater vulnerability and more widespread microvillar disruptions that affected all of the ommatidial retinula cells. The greater resistance of the eye of the male to an exposure with UV makes sense, if we consider the consequences of the retinal damage, which would clearly be a more severe handicap for an actively flying individual than for an almost sedentary one like the wingless female.

Irradiation as a methyl bromide alternative for postharvest control of Omphisa anastomosalis (Lepidoptera: Pyralidae) and Euscepes postfasciatus and Cylas formicarius elegantulus (Coleoptera: Curculionidae) in sweet potatoes.

Irradiation studies were conducted with three sweet potato, Ipomoea batatas (L.) Lam., pests to determine an effective dose for quarantine control. Dose-response tests indicated that the most radiotolerant stage occurring in roots was the pupa of sweetpotato vine borer, Omphisa anastomosalis (Guenee), and the adult of West Indian sweetpotato weevil, Euscepes postfasciatus (Fairmaire), and sweetpotato weevil, Cylas formicarius elegantulus (Summers). In large-scale confirmatory tests, irradiation of 60,000 C. formicarius elegantulus adults, 62,323 E. postfasciatus adults, and 30,282 O. anastomosalis pupae at a dose of 150 Gy resulted in no production of F1 adults, demonstrating that this dose is sufficient to provide quarantine security.

The pathogenicity of Bacillus thuringiensis ssp. kurstaki to gamma-irradiated Cadra cautella (Walker) (Lepidoptera: Phycitidae).

The present investigation deals with the effects of Bacillus thuringiensis ssp. kurstaki on various biological parameters of gamma-irradiated Cadra cautella. The pathogen, irradiation, and their combinations significantly affected the insects by increasing their mortality and developmental periods, reducing the pupal and adult survival (%), and the adult longevity and reproductive potential were also significantly reduced. It was observed that irradiation-pathogen combinations produced additive to synergistic effects on C. cautella.

Recovery from exposure to DNA-damaging chemicals in radiation-resistant insect cells.

Radioresistant TN-368 lepidopteran insect cells were examined with respect to their sensitivity to the chemical agents methyl methanesulfonate (MMS), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), propane sultone (PS), mitomycin C (MMC), and 4-nitroquinoline 1-oxide (4NQO). Based on survival ability, the TN-368 cells were more resistant than most mammalian cells to each of these agents. Concentrations of these agents which reduce survival to about 10% were used to assess recovery ability assayed by colony formation in liquid-holding and split-dose experiments. Liquid-holding experiments were performed by exposing cells in the plateau phase of growth for 1 h to 8 mM MMS, 50 microM MNNG, 9 mM PS, 110 microM MMC, or 175 microM 4NQO, removing the drug and incubating cells in spent medium for 6 h, and plating for colony formation. Split-dose experiments were performed by exposing exponentially growing cells to the above drug concentrations for 1 h, incubating in fresh medium for 6 h, exposing the cells to the agent for an additional hour, and plating. The TN-368 cells were able to significantly recover from MMS, MNNG and PS in both types of experiment. Recovery from 4NQO was observed in liquid-holding experiments and not assessed in split-dose experiments. In all cases where recovery was observed, survival enhancement was approximately 2-fold. Recovery from MMC (a cross-linking agent) exposure was not observed in either type of experiment. In addition, recovery from 8-methoxypsoralen plus UVA light (PUVA), another cross-linking treatment, was not observed. These studies indicate that DNA-DNA and/or DNA-protein crosslinking may be important molecular lesions causing death in the lepidopteran cells and that these cells may have some difficulty in repairing such damage.

Enhanced recovery from ionizing radiation damage in a lepidopteran insect cell line.

TN-368 lepidopteran insect cells display a pronounced resistance to the lethal effects of ionizing radiation and exhibit superior DNA repair capabilities. When a TN-368 cell population entering stationary growth phase is irradiated with 137Cs gamma rays and then incubated for several hours before cell dilution and plating for colony formation, the surviving fraction is increased several-fold over cells diluted and plated immediately after irradiation. Similarly, the survival of cells plated immediately following the second of two equivalent doses separated by several hours is greater than the survival of cells plated immediately following a single dose equal to the sum of the split doses. Both processes exhibit similar biphasic repair kinetics and reach maximal levels by 6 h. The phenomena appear initially to be analogous to confluent-holding and split-dose recovery as described for mammalian cells. However, the survival levels obtained for doses of 61-306 Gy after allowing for these recovery processes to occur are quite high and greatly exceed survival levels for all but relatively low doses less than 50 Gy. For example, while the survival of cells irradiated with 150 Gy is near 0.15, the survival of cells receiving 306 Gy in two equivalent split doses is approximately 0.77. Even if damage induced by the first of the split doses was completely repaired, it might be expected that the survival would be near the level of the second dose alone, or near 0.15. Instead the survival is approximately five times greater, suggesting that the first split dose stimulated a repair system not present in unirradiated cells. The situation for confluent-holding recovery is similar to that for split-dose recovery.

Enhanced survival by photoreactivation and liquid holding following UV damage of TN-368 insect cells.

These studies demonstrate that the TN-368 lepidopteran insect cell line, which is extremely resistant to the lethal effects of ionizing radiation, is also quite resistant to 254-nm ultraviolet light. While resistance to ionizing radiation in TN-368 cells has been associated with superior DNA repair processes, previous findings have indicated no correlation between survival ability and amount of unscheduled DNA synthesis in response to ultraviolet light. The present studies were undertaken to define the TN-368 ultraviolet light survival response, the ability of the cells to repair UV-induced damage by photoreactivation, the capacity of the cells to undergo UV repair during liquid holding in the dark, and the relationship between photoreactivation and liquid-holding recovery. Survival was assayed by colony formation. 254-nm irradiations were performed using germicidal lamps and photoreactivation was accomplished using black lights. Photoreactivable sectors of UV damage at 50 and 10% survival are 0.65 and 0.68, respectively. Survival responses, both with and without photoreactivation, have a small initial shoulder followed by an exponential region, and finally the curves continue to decrease but with decreasing slope. F0, Fq, and extrapolation number for the exponential portion of the curves are 77.5 J/m2, 16.8 J/m2, and 1.7 for non-photoreactivated cells and 234 J/m2, 56.1 J/m2, and 1.7 for those exposed to photoreactivating light. In the primarily exponential survival region, the fluences required to produce equivalent levels of survival in photoreactivated cells range from approximately 10.8 to 23.3 times as great as cells receiving UV light alone. The maximum survival enhancement of cells maintained under liquid-holding conditions over cells plated immediately following 100-400 J/m2 irradiations appears to be about 2-fold and occurs at 3-6 h of holding. Photoreactivation alone has a greater enhancement of survival than when photoreactivation follows liquid holding, but when liquid holding follows photoreactivation, the enhancement surpasses that of photoreactivation alone.

Structural reactions to polarized light of microvilli in photoreceptor cells of the moth Spodoptera.

Intact armyworm moths (Spodoptera exempta, Fam. Noctuidae) were illuminated by polarized monochromatic light to induce structural changes in the rhabdomeres of the compound eyes. The degree of distortion of their microvilli depends on the light energy absorbed per time unit. Under polarized light, the number of quanta absorbed varies with the position of the plane of polarization relative to the axis of the microvilli (intrinsic dichroism). Therefore, in Spodoptera, different degrees of deformations could be demonstrated in differently oriented rhabdomeres of both types of ommatidia. Moreover, in rhabdoms of the lobed type with fan-like arranged microvilli, different reactions were regularly seen in differently oriented microvilli of one rhabdomere. This indicates that microvilli may react to light individually.