Design of stable circular permutants of the GroEL chaperone apical domain.

Enhancing protein stability holds paramount significance in biotechnology, therapeutics, and the food industry. Circular permutations offer a distinctive avenue for manipulating protein stability while keeping intra-protein interactions intact. Amidst the creation of circular permutants, determining the optimal placement of the new N- and C-termini stands as a pivotal, albeit largely unexplored, endeavor. In this study, we employed PONDR-FIT's predictions of disorder propensity to guide the design of circular permutants for the GroEL apical domain (residues 191-345). Our underlying hypothesis posited that a higher predicted disorder value would correspond to reduced stability in the circular permutants, owing to the increased likelihood of fluctuations in the novel N- and C-termini. To substantiate this hypothesis, we engineered six circular permutants, positioning glycines within the loops as locations for the new N- and C-termini. We demonstrated the validity of our hypothesis along the set of the designed circular permutants, as supported by measurements of melting temperatures by circular dichroism and differential scanning microcalorimetry. Consequently, we propose a novel computational methodology that rationalizes the design of circular permutants with projected stability. Video Abstract.

Current state of students' health and factors as well as means of its improvement.

OBJECTIVE: Aim: To assess the current state of students' health in higher educational institutions in Ukraine and identify the main measures, factors, and means of its improvement. PATIENTS AND METHODS: Materials and Methods: The research was conducted in 2022-2023 and involved 266 second-year students (82 males, 184 females) from different faculties. Research methods: analysis of literary sources, observation, questionnaire surveys, and statistical methods. RESULTS: Results: A general downward trend in the level of health among students in higher educational institutions in Ukraine has been established. The deterioration of the health of male students is particularly worrying: the number of students with good health has decreased by 34.1 % over the past 10 years, while the number of students with poor and very poor health has increased by 14.0 % and 3.7 %, respectively. It has been found that 41.5 % of male and 34.2 % of female students do not know the cause of their illness and do not know how to prevent it. Students are well aware of the value of their health but do not care about it, are too lazy to exercise and rely on medications or a doctor when they are ill. CONCLUSION: Conclusions: A student's health is his or her social and personal values, level of culture, ability to withstand heavy physical and mental stress, and ability to adapt to various external factors. Therefore, the primary task of physical education departments should be to form the interests, desires, and motives of students for physical exercise.

Utilizing Extraepitopic Amino Acid Substitutions to Define Changes in the Accessibility of Conformational Epitopes of the Bacillus cereus HlyII C-Terminal Domain.

Hemolysin II (HlyII)-one of the pathogenic factors of Bacillus cereus, a pore-forming ß-barrel toxin-possesses a C-terminal extension of 94 amino acid residues, designated as the C-terminal domain of HlyII (HlyIICTD), which plays an important role in the functioning of the toxin. Our previous work described a monoclonal antibody (HlyIIC-20), capable of strain-specific inhibition of hemolysis caused by HlyII, and demonstrated the dependence of the efficiency of hemolysis on the presence of proline at position 324 in HlyII outside the conformational antigenic determinant. In this work, we studied 16 mutant forms of HlyIICTD. Each of the mutations, obtained via multiple site-directed mutagenesis leading to the replacement of amino acid residues lying on the surface of the 3D structure of HlyIICTD, led to a decrease in the interaction of HlyIIC-20 with the mutant form of the protein. Changes in epitope structure confirm the high conformational mobility of HlyIICTD required for the functioning of HlyII. Comparison of the effect of the introduced mutations on the effectiveness of interactions between HlyIICTD and HlyIIC-20 and a control antibody recognizing a non-overlapping epitope enabled the identification of the amino acid residues N339 and K340, included in the conformational antigenic determinant recognized by HlyIIC-20.

Physical literacy in Europe: The current state of implementation in research, practice, and policy.

Background/objective: The holistic concept of physical literacy (PL) embraces different person-centered qualities (physical, cognitive, affective/psychological) necessary to lead physically active lifestyles. PL has recently gained increasing attention globally and Europe is no exception. However, scientific endeavors summarizing the current state of PL in Europe are lacking. Therefore, the goal of this study was to comprehensively assess and compare the implementation of PL in research, policy, and practice across the continent. Methods: We assembled a panel of experts representing 25 European countries. Employing a complementary mixed-methods design, the experts first prepared reviews about the current state of PL in their countries (categories: research, practice/policy). The reviews underwent comparative document analysis, ensuring a transnational four-eyes principle. For re-validation purposes, the representatives completed a quantitative survey with questions reflecting the inductive themes from the document analysis. Results: The document analysis resulted in ten disjunct themes (related to "concept", "research", "practice/policy", "future/prospect") and yielded a heterogenous PL situation in Europe. The implementation state was strongly linked to conceptual discussions (e.g., existence of competing approaches), linguistic issues (e.g., translations), and country-specific traditions. Despite growing scholarly attention, PL hesitantly permeates practice and policy in most countries. Nevertheless, the experts largely anticipate increasing popularity of PL for the future. Conclusion: Despite the heterogeneous situation across Europe, the analysis has uncovered similarities among the countries, such as the presence of established yet not identical concepts. Research should intensify academic activities (conceptual-linguistic elaborations, empirical work) before PL may gain further access into practical and political spheres in the long term.

DETERMINATION OF SOMATIC, NEUROLOGICAL AND PSYCHO-EMOTIONAL MANIFESTATIONS OF THE ACUTE AND POSTCOVID PERIOD IN PATIENTS WITH A MILD COURSE OF COVID-19 IN WARTIME.

OBJECTIVE: The aim: To analyze the typical symptom complex at the stage of COVID-19 acute phase in the systemic relationship with somatic, psychosomatic, and neurological manifestations. PATIENTS AND METHODS: Materials and methods: The collection of primary material was performed by clinical-anamnestic method, laboratory, and sociological examination of patients treated out patiently. Summarizing of the results was performed according to the analysis of 100 completed cases of COVID-19 in patients aged 35-45 years (50 men and 50 women) who had no concomitant chronic pathology, and patients did not receive any vaccine dose before the disease (acute COVID-19) and during the next follow-up period (6 months). RESULTS: Results: The data of the analysis allowed us to make a grounded conclusion about the syndromic heterogeneity of COVID 19 in a standardized patients group with a mild course. CONCLUSION: Conclusions: the highest number of symptoms in the postcovid period by frequency, polymorphism, and life quality impact was found in the group of patients with subjectively tolerate acute COVID-19 most easily. Patients whose acute episode meets the mild criteria have pronounced neurological and psychoemotional manifestations during the postcovid period.

FUNCTIONAL CHARACTERISTICS OF THE CARDIOVASCULAR SYSTEM OF PATIENTS WITH ISCHEMIC HEART DISEASE WITH OBESITY.

OBJECTIVE: The aim: To determine the features of the functional characteristics of the cardiovascular system of patients with ischemic heart disease with obesity. PATIENTS AND METHODS: Materials and methods: Examined 130 persons (mostly military personnel and persons who were in the zone of active hostilities): 65 patients (the main group, 62,67±8,93 years) with coronary heart disease and obesity and 45 people of the control group (virtually healthy people, randomized by age and sex, 58,76±14,6 years). RESULTS: Results: Coronary heart disease and obesity compared to healthy individuals probably the exceed all values of the functional state of the cardiovascular system: systolic blood pressure (152.72±14.61 and 119.03±7.94 mmHg; p<0.001); diastolic blood pressure (90.74±7.36 and 80.36±6.74 mmHg; p<0.001); end-diastolic volume (103.17±40.84 and 52.48±8.58 mm3; р<0.001); end-systolic volume (47.98±29.92 and 31.47±8.42 mm3; р=0.001); end-diastolic size (4.74±0.81 and 4.12 ± 0.27 cm; р<0.001); end-systolic size (3.34±0.76 and 3.17±0.59 cm; р=0.014). CONCLUSION: Conclusions: The identified functional disorders of the heart in the comorbid course of coronary heart disease and obesity can be used for early diagnosis of cardiovascular complications in such patients and for the development of adequate therapeutic schemes.

Relationship between Changes in the Protein Folding Pathway and the Process of Amyloid Formation: The Case of Bovine Carbonic Anhydrase II.

Many proteins form amyloid fibrils only under conditions when the probability of transition from a native (structured, densely packed) to an intermediate (labile, destabilized) state is increased. It implies the assumption that some structural intermediates are more convenient for amyloid formation than the others. Hence, if a mutation affects the protein folding pathway, one should expect that this mutation could affect the rate of amyloid formation as well. In the current work, we have compared the effects of amino acid substitutions of bovine carbonic anhydrase II on its unfolding pathway and on its ability to form amyloids at acidic pH and an elevated temperature. Wild-type protein and four mutant forms (L78A, L139A, I208A, and M239A) were studied. We analyzed the change of the protein unfolding pathway by the time-resolved fluorescence technique and the process of amyloid formation by thioflavin T fluorescence assay and electron microscopy. It was revealed that I208A substitution accelerates amyloid formation and affects the structure of the late (molten globule-like)-intermediate state of carbonic anhydrase, whereas the other mutations slow down the growth of amyloids and have either no effect on the unfolding pathway (L78A, L139A) or alter the conformational states arising at the early unfolding stage (M239A).

PRACTICAL ASPECTS AND RESULTS OF COGNITIVE THERAPY IN THE EARLY RECOVERY PERIOD OF ISCHEMIC STROKE.

OBJECTIVE: The aim: To determine the impact of cognitive training on the degree of cognitive functions recovery and quality of life in the early recovery period of ischemic stroke. PATIENTS AND METHODS: Materials and methods: 108 patients with cerebral infarction were examined outpatiently, follow-up from 1 to 3 months from the onset of the disease. Basic assessment methods: screening index of cognitive disorders according to the Montreal Cognitive Assessment Scale (MoCA), SF-36 questionnaire. RESULTS: Results and Conclusions: Comprehensive rehabilitation measures for the early recovery period of ischemic stroke achieve improvement of the cognitive sphere: a significant increase in the average score on the Montreal scale of cognitive functions assessment (MoCA scale) in both observation groups.

PHYSICAL DEVELOPMENT OF STUDENTS AS AN INDICATOR OF THE PHYSICAL EDUCATION SYSTEM FUNCTIONING IN THE EDUCATIONAL INSTITUTION.

OBJECTIVE: The aim: To investigate the impact of organizational and methodological conditions of physical education system in higher educational institution on the students' physical development. PATIENTS AND METHODS: Materials and methods: The research involved 120 students of two higher educational institutions (30 men and 30 women each). Determination of physical development of students was carried out in the dynamics of learning from the first to the fourth instructional years in terms of 100 m run, 3000 m (boys) and 2000 m (girls) run, pull-ups on the crossbar (boys), push-ups (girls), 4 x 9 m shuttle run, standing long jump, lifting torso to the sitting position, torso leaning forward from the sitting position. RESULTS: Results: The research determined organizational and methodological conditions of physical education system in two educational institutions, which differ in the duration of "Physical Education" academic subject depending on the instructional year, the number of academic hours provided for the subject mastery, approaches to the basic and elective components of the curriculum, organization of fitness and health recreation as well as sports events during extracurricular activities. CONCLUSION: Conclusions: It was established that the level of students' physical development is better in the educational institutions where the physical education system functions more efficiently (compulsory physical education training sessions are held in the amount of 4 hours a week, more students are involved in educational and extracurricular physical education training sessions, where students' training motivation is higher).

Recycled ancient ghost carbonate in the Pitcairn mantle plume.

The extreme Sr, Nd, Hf, and Pb isotopic compositions found in Pitcairn Island basalts have been labeled enriched mantle 1 (EM1), characterizing them as one of the isotopic mantle end members. The EM1 origin has been vigorously debated for over 25 years, with interpretations ranging from delaminated subcontinental lithosphere, to recycled lower continental crust, to recycled oceanic crust carrying ancient pelagic sediments, all of which may potentially generate the requisite radiogenic isotopic composition. Here we find that δ26Mg ratios in Pitcairn EM1 basalts are significantly lower than in normal mantle and are the lowest values so far recorded in oceanic basalts. A global survey of Mg isotopic compositions of potentially recycled components shows that marine carbonates constitute the most common and typical reservoir invariably characterized by extremely low δ26Mg values. We therefore infer that the subnormal δ26Mg of the Pitcairn EM1 component originates from subducted marine carbonates. This, combined with previously published evidence showing exceptionally unradiogenic Pb as well as sulfur isotopes affected by mass-independent fractionation, suggests that the Pitcairn EM1 component is most likely derived from late Archean subducted carbonate-bearing sediments. However, the low Ca/Al ratios of Pitcairn lavas are inconsistent with experimental evidence showing high Ca/Al ratios in melts derived from carbonate-bearing mantle sources. We suggest that carbonate-silicate reactions in the late Archean subducted sediments exhausted the carbonates, but the isotopically light magnesium of the carbonate was incorporated in the silicates, which then entered the lower mantle and ultimately became the Pitcairn plume source.

Laser Direct Writing via Two-Photon Polymerization of 3D Hierarchical Structures with Cells-Antiadhesive Properties.

We report the design and fabrication by laser direct writing via two photons polymerization of innovative hierarchical structures with cell-repellency capability. The structures were designed in the shape of "mushrooms", consisting of an underside (mushroom's leg) acting as a support structure and a top side (mushroom's hat) decorated with micro- and nanostructures. A ripple-like pattern was created on top of the mushrooms, over length scales ranging from several µm (microstructured mushroom-like pillars, MMP) to tens of nm (nanostructured mushroom-like pillars, NMP). The MMP and NMP structures were hydrophobic, with contact angles of (127 ± 2)° and (128 ± 4)°, respectively, whereas flat polymer surfaces were hydrophilic, with a contact angle of (43 ± 1)°. The cell attachment on NMP structures was reduced by 55% as compared to the controls, whereas for the MMP, a reduction of only 21% was observed. Moreover, the MMP structures preserved the native spindle-like with phyllopodia cellular shape, whereas the cells from NMP structures showed a round shape and absence of phyllopodia. Overall, the NMP structures were more effective in impeding the cellular attachment and affected the cell shape to a greater extent than the MMP structures. The influence of the wettability on cell adhesion and shape was less important, the cellular behavior being mainly governed by structures' topography.

Bacterial Luciferases from Vibrio harveyi and Photobacterium leiognathi Demonstrate Different Conformational Stability as Detected by Time-Resolved Fluorescence Spectroscopy.

Detecting the folding/unfolding pathways of biological macromolecules is one of the urgent problems of molecular biophysics. The unfolding of bacterial luciferase from Vibrio harveyi is well-studied, unlike that of Photobacterium leiognathi, despite the fact that both of them are actively used as a reporter system. The aim of this study was to compare the conformational transitions of these luciferases from two different protein subfamilies during equilibrium unfolding with urea. Intrinsic steady-state and time-resolved fluorescence spectra and circular dichroism spectra were used to determine the stages of the protein unfolding. Molecular dynamics methods were applied to find the differences in the surroundings of tryptophans in both luciferases. We found that the unfolding pathway is the same for the studied luciferases. However, the results obtained indicate more stable tertiary and secondary structures of P. leiognathi luciferase as compared to enzyme from V. harveyi during the last stage of denaturation, including the unfolding of individual subunits. The distinctions in fluorescence of the two proteins are associated with differences in the structure of the C-terminal domain of α-subunits, which causes different quenching of tryptophan emissions. The time-resolved fluorescence technique proved to be a more effective method for studying protein unfolding than steady-state methods.

PHYSICAL STATE ASSESSMENT OF THE FIRST-YEAR FEMALE STUDENTS OF HIGHER EDUCATION INSTITUTIONS.

OBJECTIVE: The aim: Is to determine informatively significant indicators of the physical state of the first-year students of higher education institutions. PATIENTS AND METHODS: Materials and methods: The study was conducted at Prydniprovska State Academy of Civil Engineering and Architecture in 2019-2020. The study involved 100 first-year female students of the main department between the ages of 17 and 18. The state of health of female students, their motivational needs, and the level of their physical development, physical fitness, and functional state were studied. RESULTS: Results: The study summarized knowledge and experience on the problem of selecting means for assessing the physical state of students, analyzed the motivational needs and health of female students, as well as identified informative indicators of the physical state of the female students by factor analysis. CONCLUSION: Conclusions: The study indicates the need for an obligatory survey on the health of the first-year female students at the beginning of the academic year; the inclusion of the elements of modern fitness programs in the curriculum of the Physical Education discipline and the organization of sectional classes in accordance with the results of the survey if possible in order to increase students' motivation to attend classes and differentiate training in classes aimed at improving the physical state of female students.

Current state of students' health and its improvement in the process of physical education.

OBJECTIVE: The aim is to investigate the state of health of the students of Ukrainian higher education institutions. PATIENTS AND METHODS: Materials and methods: The study of state of health was conducted at Polissia National University and Zhytomyr Ivan Franko State University in 2008-2019. The results of a medical examination of 3737 students of different faculties between the ages of 17 and 22 were examined. Medical examinations were conducted by the doctors of the medical centers of universities. Totally 1208 pupils (male and female) of the schools of Zhytomyr region were involved in the questionnaire, which was conducted to analyze the attendance of physical education classes by pupils while studying at school. 648 students of Polissia National University participated in the study of the self-assessment of their own health, causes, and structure of disease incidence, the dynamics of the loss of labor capacity. RESULTS: Results: It was established that from 24 to 30.8 % of the students had diseases and a low level of physical development that indicated a low level of physical education at schools. It was revealed that the number of sick students was increasing during the study at higher educational institutions (HEI) that led to missing classes. Besides, up to 35.5 % of students had health disorders and belonged to a special medical group. And 1.7 % of the students had chronic diseases and disabilities and were exempted from sports. It was found that physical education and sports took the 5th place in the system of life values of male students and the 12th place in the system of life values of female students. CONCLUSION: Conclusions:Physical education, aimed at enhancing the functional capacity of an organism, the development of physical qualities, the improvement of the special physical and technical readiness in the sphere of physical activity types and the acquisition of professionally applied skills and abilities, is the basis for strengthening the health of students of Ukraine.

Combined use of Mössbauer spectroscopy, XPS, HRTEM, dielectric and anelastic spectroscopy for estimating incipient phase separation in lead titanate-based multiferroics.

The formation of separate phases in crystalline materials is promoted by doping with elements with different valences and ionic radii. Control of the formation of separate phases in multiferroics is extremely important for their magnetic, ferroelectric and elastic properties, which are relevant for multifunctional applications. The ordering of dopants and incipient phase separation were studied in lead titanate-based multiferroics with the formula (Pb0.88Nd0.08)(Ti0.98-xFexMn0.02)O3 (x = 0.00, 0.03, 0.04, 0.05) by means of a combination of Mössbauer spectroscopy, XPS, HRTEM, dielectric and anelastic spectroscopy. We found that Fe ions are substituted as Fe3+ at Ti sites and preferentially exhibit pentahedral coordination, whereas Ti ions have coexisting valences of Ti4+/Ti3+. Fe3+ ions are preferentially ordered in clusters, and there exists a transition temperature TC1, below which phase separation occurs between a tetragonal phase T1 free of magnetic clusters and a cubic phase, and a lower transition temperature TC2, below which the cubic phase rich in magnetic clusters is transformed into a tetragonal phase T2. The phase separation persists at the nanoscale level down to room temperature and is visible in HRTEM images as a mixing of nanodomains with different tetragonality ratios. This phase separation was observed over the whole studied concentration range of xFe values. It occurs progressively with the value of xFe, and the transition temperature TC2 decreases with the concentration from about 620 K (xFe = 0.03) to about 600 K (xFe = 0.05), while TC1 remains nearly constant.

Substitutions of Amino Acids with Large Number of Contacts in the Native State Have no Effect on the Rates of Protein Folding.

Various effects of amino acid substitutions on properties of globular proteins have been described in a large number of research papers. Nevertheless, no definite "rule" has been formulated as of yet that could be used by experimentalists to introduce desirable changes in the properties of proteins. Herein we attempt to establish such a "rule". To this end, a hypothesis is proposed on the effects of substitutions of hydrophobic residues with large number of contacts on free energies of different states of a globular protein. The hypothesis states: Substitutions of hydrophobic residues engaged in a large number of residue-residue contacts would not change the folding rate of a protein but could affect its unfolding rate. This hypothesis was verified by both theoretical and experimental analyses, generating a general rule that can facilitate the work of experimentalists on constructing mutant forms of proteins.

In vitro comparison of the effect of piezosurgery and conventional bone preparation technique on intraosseous heat generation.

The aim of this in vitro study was to compare the effect of sagittal saw handpiece with a piezoelectric device on the rise in intraosseous temperature and on the preparation time. 100 native pieces of pork ribs were cut either with S-8 S handpiece connected to Elcomed surgical motor (W&H) (n = 30) or with B6 insert connected to Piezomed (W&H) using continuous movement (n = 30) or with B6 using short breaks to perform intermittent cutting (n = 30). The rest were cut either by S-8 S (n = 5) or by B6 (n = 5) both applied by permanent pressure. The intraosseous temperature was measured by K-type thermocouple connected to digital thermometer placed in the bone 1 mm away of the cutting line. The heat generated and the time of the complete cutting were recorded. In S-8 S group the temperature never rose above 47°C. Using the B6 with permanent movement the critical temperature was reached in 16.2 ± 3.53% of the cases while taking breaks decreased the results to 2.6 ? 0.96% (p <0.001). In no cases the temperature elevation above 4700 lasted more than 60 sec. Applying the B6 by permanent pressure resulted in heat up to 90.3°C. Our results suggest that piezoelectric device could use safely according to the factory instructions, however further reduce of heat load could be achieved if the intermittent cutting motion combined with short-time cooling periods.

The major mRNP protein YB-1: structural and association properties in solution.

YB-1 is a major mRNP protein participating in the regulation of transcription and translation of a wide range of eukaryotic genes in many organisms probably due to its influence on mRNA packing into mRNPs. While the functional properties of YB-1 are extensively studied, little is known about its structural properties. In the present work we focused on studying its secondary structure, rigidity of its tertiary structure, compactness, and oligomerization in vitro by using far UV-CD, DSC, one-dimensional (1)H NMR, SAXS, sedimentation and FPLC. It was shown that only the cold shock domain within the entire YB-1 chain has a well-packed tertiary structure undergoing cooperative heat and cold denaturation transitions. In contrast, the rest of the YB-1 molecule is not rigidly packed and consists of PP II-like helical secondary structure elements and coil-like regions. At the same time, the overall dimension of the protein molecule is unexpectedly small. The polypeptide chains of YB-1 have a high tendency to form oligomers at neutral pH, while the extent and structural organization of the oligomers depend on protein concentration and ionic strength varying from compact monomeric units up to high molecular weight oligomers. These oligomers in solution are unstable and dissociate upon protein concentration decrease.

The effect of oral administration of zeolite on the energy metabolism and reproductive health of Romanian spotted breed in advanced gestation and post partum period.

The dairy cow experiences the most significant impact from negative energy balance during this period, which adversely affects reproductive health. Consequently, most pathologies affect dairy cows during this time frame. Thus, with the primary objective of reducing the incidence of these pathologies on dairy farms, we questioned whether supplemental zeolite administration in cattle feed would affect metabolism and reproductive health. Therefore, we proposed introducing an antepartum and postpartum supplementation of 400 g of zeolite in the basal diet. The control group received only the basal diet without zeolite supplementation. Monitoring the results stemmed from the consideration that reproductive health can only be present based on an unaltered energy metabolism. Hence, we deemed it necessary to analyze several metabolic markers in light of the expected outcomes concerning reproductive health. Cows treated with zeolite exhibited a calving to first service interval 12.78 days earlier than those in the control group. Moreover, the average number of services per conception used for future gestation was 0.44 lower in the zeolite-treated group compared to the control group (p<0.05). Additionally, the treatment group showed a lower presence of pathogens in the uterus and displayed a more favorable average uterine score. Observations following the completion of the research point towards an improvement in the health of transition dairy cows, opening a new path for dairy farms in terms of preventing postpartum pathologies. Indeed, the benefits from this study primarily impact the animals rather than directly influencing milk production. Therefore, further research is necessary in this regard.

A high resolution view of the fly actin cytoskeleton lacking a functional WAVE complex.

The development of multicellular organisms involves a series of morphogenetic processes coordinating a highly dynamic and organized interplay between cells and their environment. Thus, the generation of forces that drive cellular and intracellular movements is prerequisite to shape single cells into tissues and organs. The actin cytoskeleton represents a highly dynamic filamentous system providing cell structure and mechanical forces to drive membrane protrusion, cell migration and vesicle trafficking. Here, we apply the structured-illumination microscopy (SIM) technique to analyse the actin cytoskeleton in fixed Drosophila Schneider (S2R+) cells, both in wild type and in cells depleted for WAVE, a major activator of Arp2/3 mediated actin polymerization. In addition, we demonstrate that live cell SIM imaging also allows the visualization of actin-driven lamellipodial membrane dynamics at high spatial resolution in S2R+ cells. Three dimensional (3D) SIM images of up to 70 µm thick Drosophila wild-type and abi-mutant egg chambers further enables us to resolve changes of actin structures in a multicellular context with an impressive lateral and axial resolution, which is not possible with conventional confocal microscopy. Thus, the combination of superresolution 3D microscopy with Drosophila genetics and cell biology allows detailed insights into the structural and molecular requirements of different actin-dependent processes.