Evaluation of Candidate Reference Genes for Quantitative Gene Expression Analysis in Spodoptera exigu a after Long-time Exposure to Cadmium

Studies on the transcriptional control of gene expression play an important role in many areas of biology. Reference genes, which are often referred to as housekeeping genes, such as GAPDH, G3PDH, EF2, RpL7A, RpL10, TUBα and Actin, have traditionally been assumed to be stably expressed in all conditions, and they are frequently used to normalize mRNA levels between different samples in qPCR analysis. However, it is known that the expression of these genes is influenced by numerous factors, such as experimental conditions. The difference in gene expression underlies a range of biological processes, including development, reproduction and behavior. The aim of this study was to show the problems associated with using reference genes in the qPCR technique, in a study on inbred strains of Spodoptera exigua selected toward cadmium resistance. We present and discuss our results and observations, and give some recommendations concerning the use and limitations of housekeeping genes as internal standards, especially in research on insects. Our results suggest that holometabolism and poikilothermia, as well as time since metamorphosis and the level of exposure to the selective factor (cadmium in this case), have a significant effect on the expression of reference genes

Adhesion abilities and biosorption of Cd and Mg by microorganisms - first step for eco-friendly beneficiation of phosphate ore

Chemical reagents used in traditional mineral processing can be toxic and hazardous for the environment. Therefore, the use of biotechnological methods is becoming increasingly important. Great hopes are being placed in the use of microorganisms for bio-beneficiation of raw materials. However, assessment of adhesion abilities of bacteria onto minerals surface as well as biosorption of metals are essential steps before designing final process of each ore beneficiation. The main aim of this work was an investigation of biosorption of Cd and Mg, as well as adhesion abilities of five microorganism species with minerals included in the natural mixture of phosphate ore form Djebel Onk, Algeria. The ore, due to its unique composition, created conditions for adhesion of all five tested microbial strains onto apatite surface during incubation at pH 3. Moreover, Rhodococcus erythropolis CD 130, Pseudomonas fluorescens and Escherichia coli adhered distinctly onto apatite surface during incubation at pH 7. Incubation lasting 20 min at pH 4-6 created the most favorable conditions for biosorption of metals by B. subtilis and adhesion of cells. In case of C. albicans, biosorption of metals as well as adhesion of cells onto the mineral surface were more effective after longer time and in a wider pH range

Caffeine effects on AdoR mRNA expression in Drosophila melanogaster

In this study, we aimed to evaluate whether exposure to caffeine in the early stages of development affect AdoR mRNA expression levels in the fruit fly (Drosophila melanogaster) and how this will relate to the developmental success of flies. Adenosine receptors are seen as the most important biochemical targets of caffeine. Simultaneously adenosine signaling orchestrates the development and growth of insects. We demonstrate that AdoR mRNA expression in D. melanogaster is persistent from early stages till imago. Strong alterations in AdoR expression were observed in larvae that had been treated with caffeine. However, after the imaginal molt, the differences in AdoR expression between the insects from all of the test groups evened out despite a wide range of developmental success in the groups. Taken together, these results suggest that caffeine affects the expression of its cellular targets even from the early stages of fruit fly development and thus there is a significantly lower larvae-to-adult survival rate. Moreover, we also proved that the expression of AdoR undergoes a peculiar reset during the maturation of D. melanogaster despite the conditions in which larvae developed

Lagunas y obstáculos en la aplicación y el funcionamiento de los programas de administración de antimicrobianos: Resultados de una evaluación de necesidades con métodos mixtos educativos y conductuales en Francia, Estados Unidos, México e India

Antecedentes: La evidencia muestra una adherencia limitada a los principios de administración antimicrobiana (AMS). Objetivos: Identificar las lagunas educativas y las barreras sistémicas que obstaculizan la adhesión a los principios de la AMS. Métodos: Se realizó un estudio de métodos mixtos que combina un análisis temático de entrevistas cualitativas (enero-febrero de 2021) y un análisis inferencial de encuestas cuantitativas (mayo-junio de 2021). Se seleccionaron deliberadamente participantes de Francia, EE.UU., México y la India a partir de paneles en línea de profesionales de la salud para incluir a médicos infectólogos, especialistas en control de infecciones, microbiólogos clínicos, farmacólogos o farmacéuticos que se espera que apliquen los principios de la MGA en su entorno de práctica (por ejemplo, clínica, hospital académico o comunitario). Este estudio se ha guiado por un marco de análisis de carencias. Resultados: La muestra final incluyó 383 participantes (n = 33 entrevistas; n = 350 encuestas). Los resultados de los métodos mixtos indicaron que los conocimientos y habilidades de los participantes no eran óptimos para facilitar la aplicación personal y colectiva de los principios de la MGA. Los datos de la encuesta indicaron un desfase entre el conocimiento ideal y el actual de los protocolos de MGA, especialmente entre los farmacólogos (Δ0,95/4,00, p < 0,001). También se midieron las diferencias entre los niveles de conocimientos ideales y los actuales, que fueron mayores entre los especialistas en control infeccioso (Δ1,15/4,00, P < 0,001), para convencer a los directivos de los hospitales de que asignen recursos a los programas de MGA. Los obstáculos sistémicos ya existentes (por ejemplo, tiempo, financiación o formación insuficientes) se percibieron como agravados durante la pandemia COVID-19 (el 72% de los participantes en la encuesta estuvieron de acuerdo). Las deficiencias más acusadas se registraron en India y Francia. Conclusiones: Las necesidades educativas de los profesionales y los países incluidos en este estudio pueden informar las futuras actividades de desarrollo profesional continuo en AMS. Debería considerarse la posibilidad de obtener financiación adicional para abordar las barreras sistémicas percibidas. Las evaluaciones locales están justificadas para validar los resultados y la idoneidad de las intervenciones.Background: Evidence shows limited adherence to antimicrobial stewardship (AMS) principles. Objectives: To identify educational gaps and systemic barriers obstructing adherence to AMS principles. Methods: A mixed-methods study combining a thematic analysis of qualitative interviews (January-February 2021) and inferential analysis of quantitative surveys (May-June 2021) was conducted. Participants from France, the USA, Mexico and India were purposively sampled from online panels of healthcare professionals to include infectious disease physicians, infection control specialists, clinical microbiologists, pharmacologists or pharmacists expected to apply AMS principles in their practice setting (e.g. clinic, academic-affiliated or community-based hospital). A gap analysis framework guided this study. Results: The final sample included 383 participants (n = 33 interviews; n = 350 surveys). Mixed-methods findings indicated suboptimal knowledge and skills amongst participants to facilitate personal and collective application of AMS principles. Survey data indicated a gap in ideal versus current knowledge of AMS protocols, especially amongst pharmacologists (Δ0.95/4.00, P < 0.001). Gaps in ideal versus current skill levels were also measured and were highest amongst infectious control specialists (Δ1.15/4.00, P < 0.001), for convincing hospital executives to allocate resources to AMS programmes. Already existing systemic barriers (e.g. insufficient dedicated time/funding/training) were perceived as being aggravated during the COVID-19 pandemic (72% of survey participants agreed). Reported gaps were highest in India and France. Conclusions: The educational needs of professionals and countries included in this study can inform future continuous professional development activities in AMS. Additional funding should be considered to address perceived systemic barriers. Local assessments are warranted to validate results and suitability of interventions

Elemental Distribution in Reproductive and Neural Organs of the Epilachna nylanderi (Coleoptera: Coccinellidae), a Phytophage of Nickel Hyperaccumulator Berkheya coddii (Asterales: Asteraceae) by micro-PIXE

The phenomenon of metal hyperaccumulation by plants is often explained by a pathogen or herbivore defense hypothesis. However, some insects feeding on metal hyperaccumulating plants are adapted to the high level of metals in plant tissues. Former studies on species that feed on the leaves of Berkheya coddii Roessler 1958 (Asteraceae), a nickel-hyperaccumulating plant, demonstrated several protective mechanisms involved in internal distribution, immobilization, and elimination of Ni from the midgut and Malpighian tubules. These species are mainly coleopterans, including the lady beetle, Epilachna nylanderi (Mulsant 1850) (Coleoptera: Coccinellidae), collected from the ultramafic ecosystem near Barberton in South Africa. By performing particle-induced X-ray emission microanalysis elemental microanalysis (PIXE), this study examined whether Ni may be harmful to internal body systems that decide on insect reactivity (central nervous system [CNS]), their reproduction, and the relationships between Ni and other micronutrients. Data on elemental distribution of nine selected elements in target organs of E. nylanderi were compared with the existing data for other insect species adapted to the excess of metals. Micro-PIXE maps of seven regions of the CNS showed Ni mainly in the neural connectives, while cerebral ganglia were better protected. Concentrations of other bivalent metals were lower than those of Ni. Testis, compared with other reproductive organs, showed low amounts of Ni. Zn was effectively regulated at physiological dietary levels. In insects exposed to excess dietary Zn, it was also accumulated in the reproductive organs. Comparison of E. nylanderii with other insects that ingest hyperaccumulating plants, especially chrysomelid Chrysolina clathrata (Clark) (Coleoptera: Chrysomelidae), showed lower protection of the CNS and reproductive organs

Cross tolerance in beet armyworm : long-term selection by cadmium broadens tolerance to other stressors

Long lasting exposure of animals to stressing factor may lead to the selection of population able to cope with the stressor at lower cost than unexposed individuals. The aim of this study was to assess whether 130- generational selection of a beet armyworm to cadmium in food might have induced tolerance also to other stressors. The potential tolerance was assessed by means of unspecific stress markers: HSP70 concentration, DNA damage level, and energy budget indices in L5 larval instars of beet armyworm. The animals originated from Cd-exposed and control strains exposed additionally in a short-term experiment to high/low temperature or pesticide—spinosad. The application of the additional stressors caused, in general, an increase in the levels of studied parameters, in a straindependent manner. The most significant increase was found in HSP70 level in the individuals from the Cd-strain exposed to various spinosad concentration. Therefore, multigenerational contact with cadmium caused several changes that enable the insect to survive under a chronić stress, preparing the organism to the contact with an additional, new stressor. This relationship may be described as a sort of cross tolerance. This may, possibly, increase the probability of population survivorship and, at the same time, decrease the efficiency of pesticide-based plant protection efforts

Do nanoparticles cause hormesis? Early physiological compensatory response in house crickets to a dietary admixture of GO, Ag, and GOAg composite

This study aimed to identify the physiological responses of house cricket females following short-term exposure to relatively low dietary doses of graphene oxide (GO, 20 μg · g−1 food), silver (Ag, 400 μg · g−1 food) nanoparticles (NPs), or graphene oxide‑silver nanoparticle composite (GO-AgNPs, 20: 400 μg · g−1 food). Energy intake and distribution were measured on the third, sixth, and tenth day. A semi-quantitative API®ZYM assay of digestive enzyme fingerprints was performed on the third and tenth day of continuous treatment. Physicochemical properties of the NPs were obtained by combining SEM, EDX spectrometry, AFM, and DLS techniques. The obtained results showed decreased energy consumption, particularly assimilation as an early response to dietary NPs followed by compensatory changes in feeding activity leading to the same consumption and assimilation throughout the experimental period (10 days). The increased activities of digestive enzymes in NP-treated females compared to the control on the third day of the experiment suggest the onset of compensatory reactions of the day. Moreover, the insects treated with GO-AgNP composite retained more body water, suggesting increased uptake. The observed changes in the measured physiological parameters after exposure to NPs are discussed in light of hormesis

Ileal transposition in rats influenced glucose metabolism and HSP70 levels

Objective: Ileal transposition procedure (IT), in combination with sleeve gastrectomy, is widely used to induce diabetes remission and to control related metabolic abnormalities. A transposition of a long segment of distal ileum in obese Zucker rats improved glucose tolerance 6 months after IT. The premise of our study was to to examine the long - term effects of ileum transposition on the liver glycolytic enzymes content in a euglycemic group of operated Zucker rats. Methods: Twenty male Zucker rats underwent either the transposition of 50% distal ileum or a sham surgery. Six months after surgery, liver tissue concentrations of glycogen synthase kinase alpha (GSK-3α), glucose 6-phosphatase (G6PC), glycogen phosphorylase (PYGM) and phosphofructokinase (PFK) and HSP70 were assessed by immunoenzymatic methods. Results: HSP70 values were significantly higher in the IT group compared to SHAM. G6PC liver concentrations in the IT group were almost 1.45-fold lower than in the SHAM operated rats. Statistical analyses (F-test) showed HSP70 levels were significantly related to caveolin-1and SHAM group. Conclusions: Lowered glycolytic enzyme concentrations assessed in the liver suggest positive effects on glucose metabolism in long-term observations

The Structure-Properties-Cytotoxicity Interplay: A Crucial Pathway to Determining Graphene Oxide Biocompatibility

Interest in graphene oxide nature and potential applications (especially nanocarriers) has resulted in numerous studies, but the results do not lead to clear conclusions. In this paper, graphene oxide is obtained by multiple synthesis methods and generally characterized. The mechanism of GO interaction with the organism is hard to summarize due to its high chemical activity and variability during the synthesis process and in biological buffers’ environments. When assessing the biocompatibility of GO, it is necessary to take into account many factors derived from nanoparticles (structure, morphology, chemical composition) and the organism (species, defense mechanisms, adaptation). This research aims to determine and compare the in vivo toxicity potential of GO samples from various manufacturers. Each GO sample is analyzed in two concentrations and applied with food. The physiological reactions of an easy model Acheta domesticus (cell viability, apoptosis, oxidative defense, DNA damage) during ten-day lasting exposure were observed. This study emphasizes the variability of the GO nature and complements the biocompatibility aspect, especially in the context of various GO-based experimental models. Changes in the cell biomarkers are discussed in light of detailed physicochemical analysis