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Abstract Due to their complex life cycles geophytes are often neglected in conservation programs, despite they are important elements of early spring communities. Their life cycle is strongly affected by weather parameters, i.e. temperature, precipitation, and light, but the effects of these parameters are often contradictory and show high intra-annual variability even within species. Deeper knowledge about the abiotic factors affecting the population dynamics of geophytes is needed to support the designation of effective conservation plans. We aimed to explore the link between weather parameters and population dynamics of Colchicum bulbocodium, an endangered and strictly protected geophyte. We monitored three life cycle stages (flowering, growing, fruiting) of 1069 individuals in permanent plots for six consecutive years. Our results showed that life cycle of C. bulbocodium was strongly related to the actual weather parameters; the lagged effect of the previous year was weaker. Increasing temperature and lack of cold periods had negative effect on all life stages. We highlighted that population estimation based on the number of flowering individuals in a single year can underestimate population size by 40–83%. Monitoring in years following wet and cold springs and cold winters could increase the accuracy of population estimations of the flowering individuals.

Abstract Quorum sensing (QS) is a communication form between bacteria via small signal molecules that enables global gene regulation as a function of cell density. We applied a microfluidic mother machine to study the kinetics of the QS response of Pseudomonas aeruginosa bacteria to additions and withdrawals of signal molecules. We traced the fast buildup and the subsequent considerably slower decay of a population-level and single-cell-level QS response. We applied a mathematical model to explain the results quantitatively. We found significant heterogeneity in QS on the single-cell level, which may result from variations in quorum-controlled gene expression and protein degradation. Heterogeneity correlates with cell lineage history, too. We used single-cell data to define and quantitatively characterize the population-level quorum state. We found that the population-level QS response is well-defined. The buildup of the quorum is fast upon signal molecule addition. At the same time, its decay is much slower following signal withdrawal, and the quorum may be maintained for several hours in the absence of the signal. Furthermore, the quorum sensing response of the population was largely repeatable in subsequent pulses of signal molecules.

The technical difficulty of separating extracellular vesicles (EVs) from plasma proteins in human blood presents a significant hurdle in EV research, particularly during nano ultra-high-performance liquid chromatography–tandem mass spectrometric (UHPLC-MS/MS) analysis, where detecting “vesicular” proteins among abundant plasma proteins is challenging. Standardisation is a pressing issue in EV research, prompting collaborative global efforts to address it. While the MISEV guidelines offer valuable recommendations, unanswered questions remain, particularly regarding sample storage. We compared size exclusion chromatography (SEC) columns with pore sizes of 35 nm and 70 nm to identify fractions with minimal contaminating proteins and the highest concentration of small EVs (sEVs). Following column selection, we explored potential differences in the quality and quantity of sEVs isolated from platelet-free plasma (PFP) after long-term storage at −80 °C (>2.5 years) compared to freshly drawn blood. Our methodologically rigorous study indicates that prolonged storage, under correct storage and processing conditions, does not compromise sEV quality. Both columns effectively isolated vesicles, with the 70 nm column exhibiting a higher abundance of “vesicular” proteins. We propose a relatively rapid and moderately efficient protocol for obtaining a comparatively pure sEV fraction from plasma, facilitating sEV processing in clinical trials.

Abstract Carbon accounting in the land sector requires a reference level from which to calculate past losses of carbon and potential for gains using a stock-based target. Carbon carrying capacity represented by the carbon stock in primary forests is an ecologically-based reference level that allows estimation of the mitigation potential derived from protecting and restoring forests to increase their carbon stocks. Here we measured and collated tree inventory data at primary forest sites including from research studies, literature and forest inventories (7982 sites, 288,262 trees, 27 countries) across boreal, temperate, and subtropical Global Ecological Zones within Europe. We calculated total biomass carbon stock per hectare (above- and below-ground, dead biomass) and found it was 1.6 times larger on average than modelled global maps for primary forests and 2.3 times for all forests. Large trees (diameter greater than 60 cm) accounted for 50% of biomass and are important carbon reservoirs. Carbon stock foregone by harvesting of 12–52% demonstrated the mitigation potential. Estimated carbon gain by protecting, restoring and ongoing growth of existing forests equated to 309 megatons carbon dioxide equivalents per year, additional to, and higher than, the current forest sink, and comparable to the Green Deal 2030 target for carbon dioxide removals.

Abstract Aims The early diagnosis of cardiac amyloidosis (CA) is paramount, since there are effective therapies that improve patient survival. The diagnostic accuracy of classical electrocardiographic (ECG) signs, such as low voltage, pseudoinfarct pattern, and conduction disturbances in the diagnosis of CA, is inferior to that of the echocardiographic myocardial deformation criteria; therefore, our aim was to find more accurate novel ECG criteria for this purpose. Methods We tested the diagnostic value of five novel ECG criteria, two of them devised by us, in 34 patients with confirmed CA (20 transthyretin amyloidosis and 14 AL amyloidosis) and 45 control patients with left ventricular hypertrophy on echocardiography due to hypertension, valvular aortic stenosis and hypertrophic cardiomyopathy. The following novel ECG criteria, that suggested CA, were tested: QRS amplitude in lead I 4.1 (TA: 77%, SE: 93%, SP: 38%, PPV: 79%, NPV: 69%, AUC: 0.65) echocardiographic criteria. Among the classical criteria, the low voltage in limb leads criterion was present most frequently (in 73.5%) in patients with CA, with slightly worse diagnostic value than the novel ECG criteria (TA: 78.5%, SE: 73.5%, SP: 82.2%, PPV: 75.8%, NPV: 80.4%). Conclusions The novel ECG criteria [mostly the aVR

Abstract The estrus cycle in female rodents has been shown to affect a variety of physiological functions. However, little is known about its presumably thorough effect on auditory processing during the sleep–wake cycle and sleep deprivation. Vertex auditory evoked potentials (vAEPs) were evoked by single click tone stimulation and recorded during different stages of the estrus cycle and sleep deprivation performed in metestrus and proestrus in female rats. vAEPs showed a strong sleep-dependency, with the largest amplitudes present during slow wave sleep while the smallest ones during wakefulness. Higher amplitudes and longer latencies were seen in the light phase during all vigilance stages. The largest amplitudes were found during proestrus (light phase) while the shortest latencies were seen during estrus (dark phase) compared to the 2nd day diestrus baseline. High-amplitude responses without latency changes were also seen during metestrus with increased homeostatic sleep drive. More intense and faster processing of auditory information during proestrus and estrus suggesting a more effective perception of relevant environmental cues presumably in preparation for sexual receptivity. A 4-h sleep deprivation resulted in more pronounced sleep recovery in metestrus compared to proestrus without difference in delta power replacement suggesting a better tolerance of sleep deprivation in proestrus. Sleep deprivation decreased neuronal excitability and responsiveness in a similar manner both during metestrus and proestrus, suggesting that the negative consequences of sleep deprivation on auditory processing may have a limited correlation with the estrus cycle stage.

ABSTRACT Purpose: We present a technique for covering large midline loss of abdominal wall using a novel method by autologous tissues. Methods: Twenty-two patients (body mass index = 35,6 ± 6,9 kg/m2) were involved in the prospective cohort study. Acute and elective cases were included. The gap area was 450.1 ± 54 cm2. The average width of the midline gap was 16,3 ± 3,2 cm. The rectus muscles were mobilized from its posterior sheath. Both muscles were turned by180º medially, so that the complete abdominal wall gap could be covered without considerable tension. Changes in intra-abdominal pressure, quality of life and hernia recurrency were determined. Results: There was no significant increase in the intra-abdominal pressure. Wound infection and seroma occurred in four cases. Bleeding occurred in one case. Pre- and post-operative quality of life index significantly improved (23 ± 13 vs. 47 ± 6; p = 0,0013). One recurrent hernia was registered. The procedure could be performed safely and yielded excellent results. The method was applied in acute cases. The intact anatomical structure of rectus muscles was essential. Conclusions: The midline reconstruction with bilateral turned-over rectus muscles provided low tension abdominal wall status, and it did not require synthetic mesh implantation.

Abstract Most established clinical walking tests assess specific aspects of movement function (velocity, endurance, etc.) but are generally unable to determine specific biomechanical or neurological deficits that limit an individual’s ability to walk. Recently, inertial measurement units (IMU) have been used to collect objective kinematic data for gait analysis and could be a valuable extension for clinical assessments (e.g., functional walking measures). This study assesses the reliability of an IMU-based overground gait analysis during the 2-min walk test (2mWT) in individuals with spinal cord injury (SCI). Furthermore, the study elaborates on the capability of IMUs to distinguish between different gait characteristics in individuals with SCI. Twenty-six individuals (aged 22–79) with acute or chronic SCI (AIS: C and D) completed the 2mWT with IMUs attached above each ankle on 2 test days, separated by 1 to 7 days. The IMU-based gait analysis showed good to excellent test–retest reliability (ICC: 0.77–0.99) for all gait parameters. Gait profiles remained stable between two measurements. Sensor-based gait profiling was able to reveal patient-specific gait impairments even in individuals with the same walking performance in the 2mWT. IMUs are a valuable add-on to clinical gait assessments and deliver reliable information on detailed gait pathologies in individuals with SCI. Trial registration: NCT04555759.

Abstract Background Hemodialysis reactions (HDRs) are similar to complement activation-related pseudo allergy (CARPA), a hypersensitivity reaction that occurs when administering certain (nano)drugs intravenously. The pathomechanism of CARPA was described based on animal experiments. Typical CARPA-like dialysis reactions, which occur at the start of hemodialysis, have been reported using polysulfone dialyzers. However, to our knowledge, this is the first dialysis reaction that occurred towards the end of hemodialysis treatment. Case presentation This report describes a 52-year-old Caucasian male patient who had been receiving chronic hemodialysis for 3 years and exhibited a CARPA reaction during his third hour of treatment. Upon activation of the microbubble alarm, the extracorporeal system recirculated for five minutes. Following reconnection, the patient exhibited a drop in systemic blood pressure, chest pain, and dyspnea after five minutes. Symptoms disappeared spontaneously after reducing the speed of the blood pump, placing the patient in a Trendelenburg position, and administering a bolus infusion from the dialysis machine. The remaining dialysis treatment was uneventful. Conclusion Numerous case reports about reactions occurring with modern high-efficiency polysulfone dialyzers have been published. However, due to changes in the material structure by the manufacturers, we have not encountered such cases lately. The recently reported increase in thromboxane-B2 and pulmonary arterial pressure and complement activation upon re-infusion of extracorporeal blood following dialysis may explain the reaction observed here.

Abstract The parallel measurements of wavelength dependent optical absorption, particle number size distribution have made by a multi wavelength photoacoustic spectrometer (4λ-PAS) and scanning mobility particle sizer (SMPS) respectively at different modes of a diesel engine using two different types of fuel. The thermal evolution of the emission was also investigated using posterior temperature treatment of emission. The bimodal size distribution of emitted particles at a set reference temperature has been observed regardless of the applied fuel at idle. However, the emitted particulate assembly had lognormal size distribution falls into the accumulation mode at all other defined engine modes and both fuel types. The total number- and volume concentration (TNC and TVC) showed retrograde tendency with the increasing torque and rpm independently of the applied fuel types. The TNC values decreased up to 50% for both fuels with engine operation changes from idle engine mode(em#1) to low engine mode(em#2). With further increase in torque and rpm of engine, the change in TNC is negligible. On the other hand, the TVC remains more or less the same for idle to low engine mode transition and increased more than 60% for high mode (em#3) transition. The Optical Absorption Coefficient (OAC) values measured at the operational wavelengths of the 4λ-PAS instrument decreased at all wavelengths with increasing rpm and torque. The wavelength dependency quantified by Aerosol Ängström Exponent (AAE) was applied here for qualitative analysis of the carbonaceous emission and showed decreased values towards the higher engine speed and torque output of the engine. The proposed technique can be used as real-time, precise and accurate measurement of light absorption by DPM aerosols, which opens up novel possibilities for the volatility and thermal evolution investigation of diesel emissions.

The present paper gives a detailed and illustrated redescription of Entomobrya nigriventris Stach, 1929, and the description of a new species collected from open sand steppe habitat in Hungary. Based on the colour pattern, E. arenaria Winkler, Flórián & Dányi, sp. nov. is close to E. violaceolineata Stach, 1963 but differs from it by the morphology of the labral papillae and the dorsal macrochaetotaxy of the head, Th II, and Abd II–IV. The new species is also characterised by dark ventral body colouration in adult specimens. In this regard, an overview of European Entomobrya species in which the dark ventral side may occur is also provided.

Summary: The binding and function of β-arrestins are regulated by specific phosphorylation motifs present in G protein-coupled receptors (GPCRs). However, the exact arrangement of phosphorylated amino acids responsible for establishing a stable interaction remains unclear. We employ a 1D sequence convolution model trained on GPCRs with established β-arrestin-binding properties. With this approach, amino acid motifs characteristic of GPCRs that form stable interactions with β-arrestins can be identified, a pattern that we name “arreSTick.” Intriguingly, the arreSTick pattern is also present in numerous non-receptor proteins. Using proximity biotinylation assay and mass spectrometry analysis, we demonstrate that the arreSTick motif controls the interaction between many non-receptor proteins and β-arrestin2. The HIV-1 Tat-specific factor 1 (HTSF1 or HTATSF1), a nuclear transcription factor, contains the arreSTick pattern, and its subcellular localization is influenced by β-arrestin2. Our findings unveil a broader role for β-arrestins in phosphorylation-dependent interactions, extending beyond GPCRs to encompass non-receptor proteins as well.

Pharmaceuticals in the environment have emerged to a topic of global concern. Since these substances are designed to be biologically active, hazardous effects on non-target organisms are frequently reported. Here, the effects of five pharmaceuticals, one radiocontrast agent, and one degradation product on the freshwater green alga Closterium ehrenbergii were evaluated after chronic exposure of 168 h. Growth and maximum quantum yield (FV/FM) were used as endpoints and complemented by the assessment of morphology and chlorophyll fluorescence. We found that the tested antibiotics Ciprofloxacin and Ofloxacin impaired chloroplast integrity, resulting in a reduction of FV/FM from 0.1 mg/L. The disintegration of chloroplasts at higher concentrations (c = 0.3 and 0.8 mg/L, respectively) was visualized by brightfield and fluorescence microscopy. In contrast, Sulfamethoxazole interfered with cell division, leading to malformation of cells from 0.8 mg/L. Furthermore, the antibiotics exhibited a latency period of 72 h after which they started to reveal their true effects. Therefore, the importance of long-term toxicity testing is outlined in order to avoid underestimation of toxic effects of pharmaceuticals. Based on the EC10 values obtained, the antibiotics were considered to meet the criteria for classification as toxic to aquatic life with long lasting effects. The other test substances were found to exert no effects on C. ehrenbergii or only at very high concentrations and were classified as nontoxic.

Abstract AlphaFold2 (AF2) provides a 3D structure for every known or predicted protein, opening up new prospects for virtually every field in structural biology. However, working with transmembrane protein molecules pose a notorious challenge for scientists, resulting in a limited number of experimentally determined structures. Consequently, algorithms trained on this finite training set also face difficulties. To address this issue, we recently launched the TmAlphaFold database, where predicted AlphaFold2 structures are embedded into the membrane plane and a quality assessment (plausibility of the membrane-embedded structure) is provided for each prediction using geometrical evaluation. In this paper, we analyze how AF2 has improved the structural coverage of membrane proteins compared to earlier years when only experimental structures were available, and high-throughput structure prediction was greatly limited. We also evaluate how AF2 can be used to search for (distant) homologs in highly diverse protein families. By combining quality assessment and homology search, we can pinpoint protein families where AF2 accuracy is still limited, and experimental structure determination would be desirable.