The effect of ingested copper on the structural and cytotoxic properties of Steatoda grossa (Theridiidae) spider silk.

Spiders, assigned to macroconcentrators of heavy metals, are particularly threatened by the toxic effects of these chemicals. Until now, it has not been specified to what extent metals alter the processes proceeding in silk glands and if such changes could consequently influence the chemical and structural properties of the spun web threads. In the present study selected biological properties of Steatoda grossa (Theridiidae) silk yarn after nutritional exposure to copper at sublethal doses (0.234 mM CuSO4) were assessed. It was determined both changes in ultrastructure of ampullate glands and hunting web's architecture as well the cytotoxic effect in model cells (fibroblasts: line ATCC® CCL-1 NCTC clone 929), that were in contact with the analyzed biomaterial. The exposure of spiders to copper caused the occurrence of apoptotic cells in the ampullate glands as well as a significant reduction in the diameter of single fibers in double and multiple connection complexes as compared with control. At both 24 and 72 h of incubation, intensification of apoptotic and necrotic processes was observed in the fibroblast cultures that were remaining in indirect contact with the webs produced by copper-contaminated individuals. In the case of fibroblasts in direct contact with silk from the copper group, a clear cytotoxic effect resulting in an increased frequency of necrosis was observed after 72 h of incubation. The results indicated that copper may change the biological properties of spider silk and compromise its biomaterial properties.

Survival, growth and digestive functions after exposure to nanodiamonds - Transgenerational effects beyond contact time in house cricket strains.

The long-term exposure effects of nanodiamonds (NDs), spanning an organism's entire lifespan and continuing for subsequent generation, remain understudied. Most research has focused on evaluating their biological impacts on cell lines and selected organisms, typically over short exposure durations lasting hours or days. The study aimed to assess growth, mortality, and digestive functions in wild (H) and long-lived (D) strains of Acheta domesticus (Insecta: Orthoptera) after two-generational exposure to NDs in concentrations of 0.2 or 2 mg kg-1 of food, followed by their elimination in the third generation. NDs induced subtle stimulating effect that depended on the strain and generation. In the first generation, more such responses occurred in the H than in the D strain. In the first generation of H strain insects, contact with NDs increased survival, stimulated the growth of young larvae, and the activity of most digestive enzymes in mature adults. The same doses and exposure time did not cause similar effects in the D strain. In the first generation of D strain insects, survival and growth were unaffected by NDs, whereas, in the second generation, significant stimulation of those parameters was visible. Selection towards longevity appears to support higher resistance of the insects to exposure to additional stressor, at least in the first generation. The cessation of ND exposure in the third generation caused potentially harmful changes, which included, e.g., decreased survival probability in H strain insects, slowed growth of both strains, as well as changes in heterochromatin density and distribution in nuclei of the gut cells in both strains. Such a reaction may suggest the involvement of epigenetic inheritance mechanisms, which may become inadequate after the stress factor is removed.

Joint cadmium and polypropylene microparticle action in cadmium tolerant model insect.

Microplastic enlisted as a contaminant of emerging concerns in polluted environments interact with "traditional" contaminants such as metals, causing, among others, their increased accumulation in the body. Harmful effects depend on the exposed animals' possible preadaptation and/or cross-tolerance. The project aimed to assess the role of this phenomenon in the limited toxicity of polypropylene fibers (PPf) in 0%, 0.02%, 0.06, 0.18%, 0.54%, and 1.6% of Cd-supplemented food of larvae of Spodoptera exigua multigenerationally selected to cadmium tolerance. The activity of 20 digestive enzymes (API-ZYM test), defensins, and heat shock proteins, HSP70 levels in the exposed groups were used as biomarkers. PPfs caused the increase of Cd accumulation in the body, while intake of polypropylene microfibers did not change the biomarker levels. Moreover, multigenerational Cd pre-exposure, due to increased tolerance of Cd and, possibly, cross-tolerance, prepares the insects for an additional stressor (PPf) alone and in interaction with cadmium.

Hazards related to the presence of cadmium in food - Studies on the European soil centipede, Lithobius forficatus.

The soil is an environment rich in numerous potentially toxic substances/elements when present at elevated concentrations. They can be transported through the successive levels of the trophic chain. Animals living in a contaminated environment or eating contaminated food can accumulate potentially toxic elements in their bodies. One of the potentially toxic metals is cadmium, which accumulates significantly in soils. The aim of our research was to evaluate the changes caused by cadmium supplied with the food administered to invertebrates living in uncontaminated soil. The results were compared with those obtained for animals raised in contaminated soil, where cadmium entered the body via the epidermis. As the material for studies, we chose a common European soil centipede, Lithobius forficatus. Adult specimens were divided into the following experimental groups: C - control animals, Cd12 and Cd45 - animals fed with Chironomus larvae maintained in water containing 80 mg/l CdCl2, for 12 and 45 days, respectively. The material was analyzed using qualitative and quantitative analysis (transmission electron microscopy, confocal microscopy, flow cytometry, atomic absorption spectrometry). Eventually, we can conclude that the digestive system is an effective barrier against the effects of toxic metals on the entire organism, but among the gonads, ovaries are more protected than testes, however, this protection is not sufficient. Accumulation of spherites and mitochondrial alterations are probably involved in survival mechanisms of tissues after Cd intoxication.

Does age pay off? Effects of three-generational experiments of nanodiamond exposure and withdrawal in wild and longevity-selected model animals.

Nanodiamonds (NDs) are considered a material with low toxicity. However, no studies describe the effects of ND withdrawal after multigenerational exposure. The aim was to evaluate ND exposure (in the 1st and 2nd generations) effects at low concentrations (0.2 or 2 mg kg-1) and withdrawal (in the 3rd generation) in the wild (H) and longevity-selected (D) model insect Acheta domesticus. We measured selected oxidative stress parameters, immunity, types of cell death, and DNA damage. Most of the results obtained in the 1st generation, e.g., catalase (CAT), total antioxidant capacity (TAC), heat shock proteins (HSP70), defensins, or apoptosis level, confirmed no significant toxicity of low doses of NDs. Interestingly, strain-specific differences were observed. D-strain crickets reduced autophagy, the number of ROS+ cells, and DNA damage. The effect can be a symptom of mobilization of the organism and stimulation of physiological defense mechanisms in long-living organisms. The 2nd-generation D-strain insects fed ND-spiked food at higher concentrations manifested a reduction in CAT, TAC, early apoptosis, and DNA damage, together with an increase in HSP70 and defensins. ROS+ cells and cells with reduced membrane potential and autophagy did not differ significantly from the control. H-strain insects revealed a higher number of ROS+ cells and cells with reduced membrane potential, decreased CAT activity, and early apoptosis. Elimination of NDs from the diet in the 3rd generation did not cause full recovery of the measured parameters. We noticed an increase in the concentration of HSP70 and defensins (H-strain) and a decrease in apoptosis (D-strain). However, the most visible increase was a significant increase in DNA damage, especially in H-strain individuals. The results suggest prolonged adverse effects of NDs on cellular functions, reaching beyond "contact time" with these particles. Unintentional and/or uncontrolled ND pollution of the environment poses a new challenge for all organisms inhabiting it, particularly during multigenerational exposure.

Evaluation of selected biological properties of the hunting web spider (Steatoda grossa, Theridiidae) in the aspect of short- and long-term exposure to cadmium.

The study aimed at comparing the effects of short- and long-term exposure of Steatoda grossa female spiders to cadmium on the web's architecture, its energy content, and ultrastructure of ampullate glands. Simple food chain model (medium with 0.25 mM CdCl2 → Drosophila hydei flies → spider (for 4 weeks or 12 months) was used for the exposure. Analysis of Cd content provided evidence that silk fibers of the web are well protected against its incorporation irrespectively of the exposure period. Long-term exposure to cadmium resulted in the occurrence of numerous autophagosomes with degenerated organelles as well as apoptotic and necrotic cells in the ampullate glands. Concurrently, the individual silk fibers building double and multiple combination complexes were significantly thinner than in the control threads. Moreover, exposed spiders spun net with smaller mean calorific value than did the control individuals. Hence, evaluation of both the diameter of silk fibers and calorific value of the web can serve as biomarkers of the effects caused by exposure of these predators to cadmium.

Effects of food contaminated with cadmium and copper on hemocytes of Steatoda grossa (Araneae: Theridiidae).

The aim of this study was to evaluate the metabolic condition of Steatoda grossa (Theridiidae) spider, from their hemocytes, after a short-term (four-week) exposure to cadmium and copper in sublethal doses by administering them into the body of the preys. The ultrastructure of the dominant types of hemocytes, such as granulocytes, plasmatocytes and prohemocytes, was evaluated using transmission electron microscope (TEM). Quantitative evaluation of apoptotic and necrotic cells, as well as the ones with depolarized mitochondria in hemolymph, was performed using flow cytometry, while ATP concentration and ADP/ATP ratio in hemocytes were measured by luminescent methods. Cadmium, unlike copper, demonstrated proapoptotic and pronecrotic activity. Low ATP levels and high ADP/ATP ratio in hemocytes indicate a disturbance in the energy metabolism of cells and may account for their qualitative and quantitative degenerative changes. The intensification of death processes in hemocytes after an exposure to cadmium-contaminated food may impair the ability of these cells to fight infectious diseases. Copper at the applied dosage was safe for the spiders without causing visible changes in the hemocyte ultrastructure and in the level of analyzed cell death indices.

Quantitative assessment of nutrition in patients with polycystic ovary syndrome (PCOS)

Background: PCOS (polycystic ovary syndrome) is called a pathology of the XX century and affects at least 10-15% women of childbearing age. The therapy involves pharmacotherapy of hormonal imbalance, as well as the change of lifestyle, including the diet. Objective: Performing the quantitative assessment of components of diets of women with PCOS, comparing the results with current dietary standards for Polish people and defining dietary requirements for the patients. Material and Methods: The study was performed on 54 women of childbearing age (average age 26.03± 5.52) with PCOS syndrome diagnosed according to on the Rotterdam criteria. Anthropometric measurements of the patients were made and BMI and WHR calculated. Quantitative assessment of women's diets was performed based on the analysis of 3-day food diaries and food records taken from the previous 24h with the interview method. The data were introduced to a dietary software DIETA 5.0, calculating the average intake of the energy, nutrients, vitamins, minerals, cholesterol and dietary fibre. The obtained results were compared to Polish dietary guidelines. Results: Examined group was characterized by increased waist circumference (98.71± 13.6 cm) and an average WHR was 0.92± 0.08. An increased average value of BMI was also shown (28.91± 5.54 kg/m2). The patients consumed, on average, 1952.5±472.7 kcal daily, and the risk of insufficient intake of protein was determined in 36.7% of examined women. The highest risk of deficiency in minerals in women with PCOS was related to calcium (634 mg), potassium (3493 mg) and magnesium (250.1 mg), whereas with reference to vitamins deficiency as much as 70% of tested women were at risk of insufficient intake of folic acid, 36.7% of them - vitamin C, and 26.7% - vitamin B12. The average consumption of vitamin D was at the level of 3.4 µg. Test group was characterized by excessive average consumption of total fat (50%), SFA (70.4%) and saccharose (50%). The percentage of people with excessive average intake of cholesterol was at the level of 40.74%. As much as 83.3% patients consumed too low amounts of dietary fibre in their diets (<25g). Conclusions: In diet therapy of women with PCOS there should be higher intake of folic acid, vitamins D and C, cobalamin, dietary fibre and calcium. The consumption of total fats, saturated fatty acids and cholesterol should be reduced, as through facilitating the development of diabetes and cardio-vascular diseases, they affect the dysfunction of ovaries. The diet of some of the patients should be also supplemented by potassium, magnesium and zinc. The introduction of a properly balanced diet should be the key in the treatment of women with PCOS diagnosed according to Rotterdam criteria.

Fruit and vegetable consumption among the young Polish population.

Introduction: Vegetables and fruit are an essential part of a healthy and balanced diet. It has been shown that regular consumption of fruit and vegetables can reduce the risk of chronic diseases. However, in the past 8 years fruit and vegetable consumption among Polish youth has significantly decreased. Materials and methods: Patients involved in the project were participants of the Woodstock Festival Poland 2014. During three days of the festival, 615 young adults (18­35 years) were enrolled in the study. The study participants had to fill in a questionnaire that included questions about the frequency of fruit and vegetable consumption, portion sizes, and knowledge of the recommended daily intake. We also measured basic anthropometric parameters. Results: The data from our study showed that 97.6% of respondents consumed fewer than 4 servings of vegetables and fruit. In addition 39% of them ate 2 or fewer vegetable servings per day. We found that men ate significantly less fruit and fewer vegetables than women (p < 0.05). Additionally, almost 75% of men responded that proper consumption of fruit and vegetables should be at the level of 2­3 servings, and only 22.3% of them said that the proper level is 4­5 servings. Conclusions: We need to keep in mind that only clear information about vegetables and fruit, as well as highlighting the importance of the principles of nutrition, can give effective results in the future. It also appears that nutritional education should be conducted not only at the early stages of development but also in adolescents.

DNA damage in haemocytes and midgut gland cells of Steatoda grossa (Theridiidae) spiders exposed to food contaminated with cadmium.

The aim of this study was to assess the genotoxic effects of Cd on haemocytes and midgut gland cells of web-building spiders, Steatoda grossa (Theridiidae), exposed to the metal under laboratory conditions. Analyzes were conducted on adult females and males, fed for four weeks with cadmium-contaminated Drosophila hydei flies, grown on a medium suplemented with 0.25 mM CdCl2. The comet assay, providing a quantitative measure of DNA strand breaks, was used to evaluate the DNA damage caused by the metal. Cadmium content was measured in whole spider bodies by the AAS method. Metal body burden was significantly lower in females (0.25 µgg(-1) dry weight) than in males (3.03 µgg(-1) dry weight), suggesting that females may have more effective mechanisms controlling the uptake of metal, via the digestive tract, or its elimination from the body. Irrespectively of sex, spiders fed prey contaminated with cadmium showed significantly higher values of comet parameters: tail DNA (TDNA), tail length (TL) and olive tail moment (OTM), in comparison with the control. In midgut gland cells, the level of DNA damage was higher for males than females, while in haemocytes the genotoxic effect of cadmium was greater in females. The obtained results indicate that in spiders cadmium displays strong genotoxic effects and may cause DNA damage even at low concentrations, however the severity of damage seems to be sex- and internal organ-dependent. The comet assay can be considered a sensitive tool for measuring the deleterious effect of cadmium on DNA integrity in spiders.