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Imaging biological macromolecules in their native state with single-particle cryo-electron microscopy (cryo-EM) or in situ cryo-electron tomography (cryo-ET) requires optimized approaches for the preparation and vitrification of biological samples. Here, we describe EasyGrid, a versatile technology enabling systematic, tailored and advanced sample preparation for cellular and structural biology. This automated, standalone platform combines in-line plasma treatment, microfluidic dispensing, blot-less sample spreading, jet-based vitrification and on-the-fly grid quality control using light interferometry to streamline cryo-EM sample optimization. With EasyGrid, we optimized grid preparation for different purified macromolecular complexes and subsequently determined their structure with cryo- EM. We also demonstrated how the platform allows better vitrification of large, mammalian cells compared to standard plunge-freezing. Automated sample preparation with EasyGrid establishes an advanced, high-throughput platform for both single-particle cryo-EM and cellular cryo-ET sample preparation.Competing Interest StatementGergely Papp, Florent Cipriani - pending patent WO 2020/058140 Gergely Papp - European patent application 23 209 700.6

The permanently growing interest in amateur and professional sports activities among young, middle-aged and elderly athletes raises serious concerns about athletes' health, the safety of physical training, and the sports-related risk of sudden cardiac death and other cardiovascular complications during exercise. In recent years there has been an increasing number of cases of sudden cardiac death during physical effort. At present, life expectancy in the most developed countries grows rapidly and the number of people >65 years dramatically increases. Moreover, biological age of the population is lower. Subsequently, relevant increase in the number of elderly athletes involved in various types of sports activities has been reported in many countries. It was also demonstrated that physical activity has strong beneficial effect on cognitive functions, psychomotor performance and thus exercise capacity, which is very important for the elderly people in their everyday routine activities. Nonetheless, it should be remembered that participation in amateur and professional sports activities may be associated with risk of serious cardiovascular events in the elderly athletes often suffering from various civilization diseases. It is also reported that the number of elderly people after interventional cardiology procedures, open heart surgery and cancer treatment (chemotherapy, radiotherapy), who wish to participate in various sports activities, systematically grows. The authors of the paper conduct a literature review on cardiovascular risk assessment in the elderly athletes including the contemporary cardiology diagnostic methods and diagnostic schemes to prevent sudden cardiac death and other cardiovascular events during exercise. The controversies over efficacy of particular diagnostic tools to detect cardiovascular diseases in the elderly athletes and worldwide epidemiologic data concerning risk of sudden cardiac death during physical exercise have been also presented. Here, the authors have derived suggestions for establishment of comprehensive diagnostic schemes to prevent sudden cardiac death during sports activities. Med Pr Work Health Saf. 2024;75(3):233-241

Background Eliminating hepatitis C virus (HCV) infections is a goal set by the World Health Organization. This has become possible with the introduction of highly effective and safe direct-acting antivirals (DAA) but limitations remain due to undiagnosed HCV infections and loss of patients from the cascade of care at various stages, including those lost to follow-up (LTFU) before the assessment of the effectiveness of the therapy. The aim of our study was to determine the extent of this loss and to establish the characteristics of patients experiencing it. Methods Patients with chronic HCV infection from the Polish retrospective multicenter EpiTer-2 database who were treated with DAA therapies between 2015 and 2023 were included in the study. Results In the study population of 18,968 patients, 106 had died by the end of the 12-week post-treatment follow-up period, and 509 patients did not report for evaluation of therapy effectiveness while alive and were considered LTFU. Among patients with available assessment of sustained virological response (SVR), the effectiveness of therapy was 97.5%. A significantly higher percentage of men ( p <0.0001) and a lower median age ( p =0.0001) were documented in LTFU compared to the group with available SVR assessment. In LTFU patients, comorbidities such as alcohol ( p <0.0001) and drug addiction ( p =0.0005), depression ( p =0.0449) or other mental disorders ( p <0.0001), and co-infection with human immunodeficiency virus (HIV) ( p <0.0001) were significantly more common as compared to those with SVR assessment. They were also significantly more often infected with genotype (GT) 3, less likely to be treatment-experienced and more likely to discontinue DAA therapy. Conclusions In a real-world population of nearly 19,000 HCV-infected patients, we documented a 2.7% loss to follow-up rate. Independent predictors of this phenomenon were male gender, GT3 infection, HIV co-infection, alcohol addiction, mental illnesses, lack of prior antiviral treatment and discontinuation of DAA therapy.

Background The development of research on exercise physiology in the last several years has allowed to better understand mechanisms affecting exercise performance in athletes. Despite this, the causes of exercise intolerance in athletes are not fully elucidated. The exercise performance is a complex process and regardless of the proper functioning of the cardiovascular system may be subject to significant changes under the influence of nervous system and thermoregulatory mechanisms. The purpose of this work is to assess the relationship between psychomotor performance ( sprawność psychomotoryczna – SP), core body temperature (Tc) and exercise performance parameters as measured by spiroergometry (cardiopulmonary exercise testing – CPX) and the diagnostic accuracy of neurothermometabolic index (NTMI) as a new marker of exercise performance in athletes. Material and Methods Forty-four healthy athletes participated in the study. Each of the examined athletes underwent CPX for the assessment of ventilatory parameters (maximal oxygen consumption – VO 2 max, minute ventilation – VE, ventilatory equivalent for carbon dioxide at anaerobic threshold – VECO2AT, maximal carbon dioxide output – VCO 2 and anaerobic threshold), SP tests (average reaction time to light stimulus, the number of correctly received light stimuli, number of missed light stimuli) before and immediately after CPX. In addition, Tc was continually measured during CPX. Followingly, NTMI was calculated using Tc during the maximum exercise, oxygen consumption at anaerobic threshold and average reaction time to light stimulus as measured after CPX. Results A significant relationship was found between Tc during the maximum effort in CPX and SP after CPX. A significant relationship between SP before CPX and VECO 2 AT during CPX was also confirmed. Psychomotor performance shortly after CPX was also significantly related to VCO 2 . In addition, NTMI was significantly associated with CPX parameters. Conclusions The study results confirmed the relevant relationship between SP, Tc during the maximum exercise and ventilatory parameters as measured by CPX in athletes. Neurothermometabolic index is a reliable marker of exercise performance in athletes. Med Pr Work Health Saf. 2025;76(3):179–192

Background Anxiety disorders can be an independent risk factor of many somatic conditions and complication of various organic states leading to the unfavourable course of the primary disease and poor prognosis. The main goal of the study was to evaluate anxiety reactions with regard to the disturbances of homeostatic processes within respiratory system, from which various somatic symptoms were reported. Material and Methods The study group I comprised 50 subjects (aged M±SD 62.5±2.29 years) presenting with generalized anxiety disorders, and 60 normal subjects (aged M±SD 61.94±2.32 years), volunteers as a reference group II. In all the study subjects spirometry and pulse oxymetry were performed. A level of declared dyspnoea was assessed using the 0–10 Borg scale. Moreover, all persons were subjected to physical examination after taking medical history. Results In the group I, hyperventilation was reported in the chronic form, and a declared dyspnoea level was relatively low, despite the proper oxygen saturation (SaO 2 ) of arterial blood. The objective analysis of the Tiffeneau index showed obturative ventilation disturbances, and evaluation of forced expiratory volume in 1 sec (FEV 1 ) correspondet to obturation of a mild type (FEV 1 = 80.88% of normal value) and a significant decrease in inspiration vs. expiration parameters (e.g., forced inspiratory vital capacity to forced expiratory vital capacity) that indicated the inspiration type of ventilation impairment. In turn, the analysis of the maximal expiratory flow values showed that obturation in all persons from the group I was located mainly in the upper respiratory tract. The statistically significant correlations between the declared dyspnoea level and spirometry parameters in the group I were also noted. In contrast, subjects from group II did not declare dyspnea. Conclusions It was shown that anxiety disorders affect not only a breathing pattern but also can induce the objective changes of spirometric parameters like in the organic diseases. As the result of erroneous interpretation of afferent stimuli in the respiratory centre, the normal SaO 2 can be a stimulus triggering dyspnoea. The characteristics accompanying a feeling of dyspnoea suggested the psychogenic background of the symptoms. Med Pr Work Health Saf. 2025;76(3):167–177

Background: The development of research on exercise physiology in the last several years has allowed to better understand mechanisms affecting exercise performance in athletes. Despite this, the causes of exercise intolerance in athletes are not fully elucidated. The exercise performance is a complex process and regardless of the proper functioning of the cardiovascular system may be subject to significant changes under the influence of nervous system and thermoregulatory mechanisms. The purpose of this work is to assess the relationship between psychomotor performance (sprawnosc psychomotoryczna--SP), core body temperature (Tc) and exercise performance parameters as measured by spiroergometry (cardiopulmonary exercise testing--CPX) and the diagnostic accuracy of neurothermometabolic index (NTMI) as a new marker of exercise performance in athletes. Material and Methods: Forty-four healthy athletes participated in the study. Each of the examined athletes underwent CPX for the assessment of ventilatory parameters (maximal oxygen consumption--V[O.sub.2]max, minute ventilation--VE, ventilatory equivalent for carbon dioxide at anaerobic threshold--VEC[O.sub.2]AT, maximal carbon dioxide output-- VC[O.sub.2] and anaerobic threshold), SP tests (average reaction time to light stimulus, the number of correctly received light stimuli, number of missed light stimuli) before and immediately after CPX. In addition, Tc was continually measured during CPX. Followingly, NTMI was calculated using Tc during the maximum exercise, oxygen consumption at anaerobic threshold and average reaction time to light stimulus as measured after CPX. Results: A significant relationship was found between Tc during the maximum effort in CPX and SP after CPX. A significant relationship between SP before CPX and VEC[O.sub.2]AT during CPX was also confirmed. Psychomotor performance shortly after CPX was also significantly related to VC[O.sub.2]. In addition, NTMI was significantly associated with CPX parameters. Conclusions: The study results confirmed the relevant relationship between SP, Tc during the maximum exercise and ventilatory parameters as measured by CPX in athletes. Neurothermometabolic index is a reliable marker of exercise performance in athletes. Med Pr Work Health Saf. 2025;76(3):179-192 Key words: exercise performance, central nervous system, athletes, psychomotor performance, core body temperature, spiroergometry Wstep: Rozwoj badan nad fizjologia wysilku w ostatnich kilkunastu latach pozwolil na lepsze zrozumienie mechanizmow wplywajacych na wydolnosc fizyczna u sportowcow. Mimo to przyczyny nietolerancji wysilku fizycznego u sportowcow nie sa w pelni jasne. Wydolnosc fizyczna jest procesem zlozonym i niezaleznie od prawidlowego funkcjonowania ukladu sercowo- naczyniowego moze podlegac istotnym zmianom pod wplywem ukladu nerwowego oraz mechanizmow termoregulacyjnych. Celem niniejszej pracy jest ocena zwiazkow pomiedzy sprawnoscia psychomotoryczna (SP) i temperatura wewnetrzna ciala (core body temperature--Tc) a wybranymi parametrami wydolnosciowymi w tescie spiroergometrycznym (sercowo-plucny test wysilkowy, cardiopulmonary exercise testing-- CPX) oraz wartosci diagnostycznej wskaznika neurotermometabolicznego (neurothermometabolic index--NTMI) jako nowego markera wydolnosci fizycznej u sportowcow. Material i metody: W badaniu uczestniczylo 44 zdrowych sportowcow. U wszystkich przeprowadzono CPX w celu oceny parametrow wentylacyjnych, tj. maksymalnego poboru tlenu przez tkanki (maximum oxygen consumption--V[O.sub.2]max), wentylacji minutowej (minute ventilation--VE), ekwiwalentu wentylacyjnego dla dwutlenku wegla na progu beztlenowym (ventilatory equivalent for carbon dioxide at anaerobic threshold--VECO2AT), wydalania dwutlenku wegla podczas maksymalnego wysilku (maximal carbon dioxide output--VC[O.sub.2]) i progu beztlenowego, ocene SP (sredni czas reakcji zlozonej na bodziec swietlny, liczba poprawnych i blednych odpowiedzi na bodziec swietlny) przed i bezposrednio po CPX oraz pomiar Tc w CPX. Na podstawie uzyskanych wartosci Tc podczas maksymalnego wysilku, wartosci poboru tlenu na progu beztlenowym i sredniego czasu reakcji na bodziec swietlny zmierzonego po CPX wyznaczono NTMI. Wyniki: Stwierdzono istotny zwiazek pomiedzy Tc podczas maksymalnego wysilku a SP oceniana zaraz po zakonczeniu CPX. Potwierdzono istotny zwiazek pomiedzy SP przed CPX a VEC[O.sub.2]AT w CPX. Sprawnosc psychomotoryczna oceniona po zakonczeniu CPX byla istotnie zwiazana z VC[O.sub.2]. Wskaznik neurotermometaboliczny byl istotnie zwiazany z V[O.sub.2]max, VE i VC[O.sub.2] w CPX. Wnioski: Wyniki badan potwierdzily istotny zwiazek pomiedzy SP, obciazeniem termicznym i parametrami wentylacyjnymi w CPX u sportowcow. Wskaznik neurotermometaboliczny jest wartosciowym markerem wydolnosci fizycznej u sportowcow. Med Pr Work Health Saf. 2025;76(3):179-192 Slowa kluczowe: wydolnosc fizyczna, osrodkowy uklad nerwowy, sportowcy, sprawnosc psychomotoryczna, temperatura wewnetrzna ciala, spiroergometria

Background: Anxiety disorders can be an independent risk factor of many somatic conditions and complication of various organic states leading to the unfavourable course of the primary disease and poor prognosis. The main goal of the study was to evaluate anxiety reactions with regard to the disturbances of homeostatic processes within respiratory system, from which various somatic symptoms were reported. Material and Methods: The study group I comprised 50 subjects (aged M [+ or -] SD 62.5 [+ or -] 2.29 years) presenting with generalized anxiety disorders, and 60 normal subjects (aged M [+ or -] SD 61.94 [+ or -] 2.32 years), volunteers as a reference group II. In all the study subjects spirometry and pulse oxymetry were performed. A level of declared dyspnoea was assessed using the 0-10 Borg scale. Moreover, all persons were subjected to physical examination after taking medical history. Results: In the group I, hyperventilation was reported in the chronic form, and a declared dyspnoea level was relatively low, despite the proper oxygen saturation (Sa[O.sub.2]) of arterial blood. The objective analysis of the Tiffeneau index showed obturative ventilation disturbances, and evaluation of forced expiratory volume in 1 sec ([FEV.sub.1]) correspondet to obturation of a mild type ([FEV.sub.1] = 80.88% of normal value) and a significant decrease in inspiration vs. expiration parameters (e.g., forced inspiratory vital capacity to forced expiratory vital capacity) that indicated the inspiration type of ventilation impairment. In turn, the analysis of the maximal expiratory flow values showed that obturation in all persons from the group I was located mainly in the upper respiratory tract. The statistically significant correlations between the declared dyspnoea level and spirometry parameters in the group I were also noted. In contrast, subjects from group II did not declare dyspnea. Conclusions: It was shown that anxiety disorders affect not only a breathing pattern but also can induce the objective changes of spirometric parameters like in the organic diseases. As the result of erroneous interpretation of afferent stimuli in the respiratory centre, the normal Sa[O.sub.2] can be a stimulus triggering dyspnoea. The characteristics accompanying a feeling of dyspnoea suggested the psychogenic background of the symptoms. Med Pr Work Health Saf. 2025;76(3):167-177 Key words: generalized anxiety disorders, older adults, spirometry parameter, declared dyspnoea, hyperventilation, respiratory pattern generator Wstep: Zaburzenia lekowe moga stanowic niezalezny czynnik ryzyka wielu chorob somatycznych oraz wystepowac jako powiklanie chorob organicznych, bedac przyczyna ich niekorzystnego przebiegu i gorszego rokowania. Glownym celem badan byla ocena wplywu reakcji lekowych na zaburzenia procesow homeostatycznych w obrebie ukladu oddechowego, ze strony ktorego zglaszano liczne objawy podmiotowe. Material i metody: Badaniami objeto grupe 50 osob z zaburzeniami lekowymi uogolnionymi w wieku M [+ or -] SD 62,5 [+ or -] 2,29 roku (grupa I) oraz 60 zdrowych ochotnikow w wieku M [+ or -] SD 61,94 [+ or -] 2,32 roku (grupa II) jako grupe odniesienia. U osob z obu grup przeprowadzono badanie spirometryczne i pulsoksymetrie. Poziom deklarowanej dusznosci oceniono za pomoca 10-stopniowej skali Borga. Ponadto od wszystkich uczestnikow zebrano wywiad lekarski i przeprowadzano u nich badanie fizykalne. Wyniki: W grupie I hiperwentylacja wystepowala w postaci przewleklej, a poziom deklarowanej dusznosci odpowiadal dusznosci o slabym nasileniu, pomimo prawidlowej saturacji krwi tetniczej tlenem (oxygen saturation--Sa[O.sub.2]). Obiektywna ocena wskaznika Tiffeneau wykazala obturacyjne zaburzenia wentylacji, a ocena natezonej objetosci wydechowej w czasie 1 s (forced expiratory volume in 1 s--[FEV.sub.1]) odpowiadala obturacji lagodnej ([FEV.sub.1] = 80,88% wartosci naleznej), natomiast spadek wartosci parametrow wdechowych w stosunku do ich wydechowych odpowiednikow, np. natezonej pojemnosci zyciowej wdechowej do natezonej pojemnosci zyciowej wydechowej, wskazywal na zaburzenia typu wdechowego. Z kolei analiza procentowa wartosci naleznych maksymalnych przeplywow wydechowych w grupie I pozwolila ustalic, ze obturacja wystepowala glownie na poziomie gornych drog oddechowych. Odnotowano takze statystycznie istotne korelacje miedzy poziomem deklarowanej dusznosci a parametrami spirometrycznymi u osob z grupy I, podczas gdy te z grupy II nie deklarowaly dusznosci. Wnioski: Wykazano, ze zaburzenia lekowe, poza wplywem na zmiane wzorca oddechowego, moga wywolywac obiektywne zmiany parametrow spirometrycznych na rowni z chorobami organicznymi. W wyniku blednej interpretacji bodzcow aferentnych w osrodku oddechowym prawidlowa Sa[O.sub.2] mogla byc bodzcem wyzwalajacym dusznosc, co lacznie z jej charakterystycznymi cechami przemawialo za psychogennym podlozem tej dusznosci. Med Pr Work Health Saf. 2025;76(3):167-177 Slowa kluczowe: zaburzenie lekowe uogolnione, osoby starsze, parametry spirometryczne, deklarowana dusznosc, hiperwentylacja, generator wzorca oddechowego

The clinical and metabolic interactions between hepatitis C virus (HCV) infection and diabetes mellitus (DM) are well documented. The study aimed to compare HCV-infected patients with and without DM. The analysis included 18,968 patients treated with direct-acting antivirals (DAAs) between 2015 and 2023, whose data were collected retrospectively. In the study population, 2179 patients (11.5%) were diagnosed with DM. Compared to the non-diabetic population, they were male-dominated ( p  = 0.003), had a significantly higher proportion of patients aged ≥ 50 years ( p  < 0.001), and were more burdened with comorbidities ( p  < 0.001). The most common HCV genotype was 1b with a significantly higher prevalence in the diabetic group ( p  < 0.001). Liver disease advancement was higher in diabetic patients, with 17.9% advanced fibrosis and 48% cirrhosis compared to 13.2% ( p  < 0.001) and 21.8% ( p  < 0.001) in the non-diabetic population. The effectiveness of DAA therapy in patients with DM was significantly lower compared to the population without diabetes, both in intent-to-treat analysis 93.1% vs. 94.6%, p  = 0.015, and per-protocol analysis 96.8% vs. 97.7%, p  = 0.0128, however, logistic regression analysis did not confirm the role of diabetes as an independent predictor of treatment failure, suggesting that in the absence of other negative prognostic factors, DM alone does not reduce the chances of cure.