Biphasic decay kinetics suggest progressive slowing in turnover of latently HIV-1 infected cells during antiretroviral therapy

BACKGROUND: Mathematical models based on kinetics of HIV-1 plasma viremia after initiation of combination antiretroviral therapy (cART) inferred HIV-infected cells to decay exponentially with constant rates correlated to their strength of virus production. To further define in vivo decay kinetics of HIV-1 infected cells experimentally, we assessed infected cell-classes of distinct viral transcriptional activity in peripheral blood mononuclear cells (PBMC) of five patients during 1 year after initiation of cART RESULTS: In a novel analytical approach patient-matched PCR for unspliced and multiply spliced viral RNAs was combined with limiting dilution analysis at the single cell level. This revealed that HIV-RNA+ PBMC can be stratified into four distinct viral transcriptional classes. Two overlapping cell-classes of high viral transcriptional activity, suggestive of a virion producing phenotype, rapidly declined to undetectable levels. Two cell classes expressing HIV-RNA at low and intermediate levels, presumably insufficient for virus production and occurring at frequencies exceeding those of productively infected cells matched definitions of HIV-latency. These cells persisted during cART. Nevertheless, during the first four weeks of therapy their kinetics resembled that of productively infected cells. CONCLUSIONS: We have observed biphasic decays of latently HIV-infected cells of low and intermediate viral transcriptional activity with marked decreases in cell numbers shortly after initiation of therapy and complete persistence in later phases. A similar decay pattern was shared by cells with greatly enhanced viral transcriptional activity which showed a certain grade of levelling off before their disappearance. Thus it is conceivable that turnover/decay rates of HIV-infected PBMC may be intrinsically variable. In particular they might be accelerated by HIV-induced activation and reactivation of the viral life cycle and slowed down by the disappearance of such feedback-loops after initiation of cART

Indoor environment in retrofitted offices equipped with radiant ceiling panels

U radu se ocjenjuje unutarnja okolina renoviranih ureda koji su, kao pokusni primjer, opremljeni naprednim stropnim isijavajućim pločama koje se primjenjuju za grijanje i hlađenje. Obavljene su opsežne analize na bazi raznih nizova stvarnih mjerenja kako bi se ocijenila djelotvornost i prikladnost predložene koncepcije za adaptiranje prostora. Ustanovljeno je da čitav sustav za temperiranje uredskih prostora funkcionira na prikladan način, te da raspon unutarnjih temperatura zraka omogućava postizanje najviše razine udobnosti.The indoor environment in retrofitted pilot demonstration offices, equipped with advanced radiant ceiling panels that were used for heating and cooling, is evaluated in the paper. Comprehensive analyses based on various sets of real case measurements were carried out in order to assess the efficiency and adequacy of the proposed retrofitting concept. It was established that the entire system for tempering the offices operates well, and that the range of indoor air temperatures ensures the highest level of comfort

Duration of invasive mechanical ventilation prior to extracorporeal membrane oxygenation is not associated with survival in acute respiratory distress syndrome caused by coronavirus disease 2019

BACKGROUND: Duration of invasive mechanical ventilation (IMV) prior to extracorporeal membrane oxygenation (ECMO) affects outcome in acute respiratory distress syndrome (ARDS). In coronavirus disease 2019 (COVID-19) related ARDS, the role of pre-ECMO IMV duration is unclear. This single-centre, retrospective study included critically ill adults treated with ECMO due to severe COVID-19-related ARDS between 01/2020 and 05/2021. The primary objective was to determine whether duration of IMV prior to ECMO cannulation influenced ICU mortality. RESULTS: During the study period, 101 patients (mean age 56 [SD ± 10] years; 70 [69%] men; median RESP score 2 [IQR 1–4]) were treated with ECMO for COVID-19. Sixty patients (59%) survived to ICU discharge. Median ICU length of stay was 31 [IQR 20.7–51] days, median ECMO duration was 16.4 [IQR 8.7–27.7] days, and median time from intubation to ECMO start was 7.7 [IQR 3.6–12.5] days. Fifty-three (52%) patients had a pre-ECMO IMV duration of > 7 days. Pre-ECMO IMV duration had no effect on survival (p = 0.95). No significant difference in survival was found when patients with a pre-ECMO IMV duration of < 7 days (< 10 days) were compared to ≥ 7 days (≥ 10 days) (p = 0.59 and p = 1.0). CONCLUSIONS: The role of prolonged pre-ECMO IMV duration as a contraindication for ECMO in patients with COVID-19-related ARDS should be scrutinised. Evaluation for ECMO should be assessed on an individual and patient-centred basis. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13613-022-00980-3

LRRK2 protein levels are determined by kinase function and are crucial for kidney and lung homeostasis in mice

Mutations in leucine-rich repeat kinase 2 (LRRK2) cause late-onset Parkinson's disease (PD), but the underlying pathophysiological mechanisms and the normal function of this large multidomain protein remain speculative. To address the role of this protein in vivo, we generated three different LRRK2 mutant mouse lines. Mice completely lacking the LRRK2 protein (knock-out, KO) showed an early-onset (age 6 weeks) marked increase in number and size of secondary lysosomes in kidney proximal tubule cells and lamellar bodies in lung type II cells. Mice expressing a LRRK2 kinase-dead (KD) mutant from the endogenous locus displayed similar early-onset pathophysiological changes in kidney but not lung. KD mutants had dramatically reduced full-length LRRK2 protein levels in the kidney and this genetic effect was mimicked pharmacologically in wild-type mice treated with a LRRK2-selective kinase inhibitor. Knock-in (KI) mice expressing the G2019S PD-associated mutation that increases LRRK2 kinase activity showed none of the LRRK2 protein level and histopathological changes observed in KD and KO mice. The autophagy marker LC3 remained unchanged but kidney mTOR and TCS2 protein levels decreased in KD and increased in KO and KI mice. Unexpectedly, KO and KI mice suffered from diastolic hypertension opposed to normal blood pressure in KD mice. Our findings demonstrate a role for LRRK2 in kidney and lung physiology and further show that LRRK2 kinase function affects LRRK2 protein steady-state levels thereby altering putative scaffold/GTPase activity. These novel aspects of peripheral LRRK2 biology critically impact ongoing attempts to develop LRRK2 selective kinase inhibitors as therapeutics for PD

Rif1 maintains telomeres and mediates DNA repair by encasing DNA ends

In yeast, Rif1 is part of the telosome, where it inhibits telomerase and checkpoint signaling at chromosome ends. In mammalian cells, Rif1 is not telomeric, but it suppresses DNA end resection at chromosomal breaks, promoting repair by nonhomologous end joining (NHEJ). Here, we describe crystal structures for the uncharacterized and conserved ∼125-kDa N-terminal domain of Rif1 from Saccharomyces cerevisiae (Rif1-NTD), revealing an α-helical fold shaped like a shepherd's crook. We identify a high-affinity DNA-binding site in the Rif1-NTD that fully encases DNA as a head-to-tail dimer. Engagement of the Rif1-NTD with telomeres proved essential for checkpoint control and telomere length regulation. Unexpectedly, Rif1-NTD also promoted NHEJ at DNA breaks in yeast, revealing a conserved role of Rif1 in DNA repair. We propose that tight associations between the Rif1-NTD and DNA gate access of processing factors to DNA ends, enabling Rif1 to mediate diverse telomere maintenance and DNA repair functions

Effects of specific parameters on simulations of energy use and air temperatures in offices equipped with radiant heating/cooling panels

When creating a simulation model to assess the performance of buildings, there is usually a lack of feedback information. Only in the case of measurements of a real building is a direct comparison of the measured values and simulated results possible. Parameter data related to users’ behavior or other events can also be obtained. Their evaluated frequency, magnitude and duration, along with boundary conditions, are crucial for the results. It is clear that none of them can be predicted very accurately. Most of them, however, are needed for computer modeling. In this paper we analyzed the well-defined TRNSYS simulation model of offices equipped with radiant ceiling panels for heating and cooling. The model was based on real case offices and was validated based on measurements for 1 year. The analysis included simulations in order to define what effect the parameters related mainly to users have on the energy use and the indoor air temperatures. The study confirmed that specific human activities influence the annual energy use to a relatively small degree and that their effects often counteract. It also confirmed the even more important fact that although small, these activities can influence the thermal comfort of users. It is believed that despite the fact that this research was based on an analysis of offices equipped with radiant ceiling panels, most of the results could be applied generally

Post-fire effects on development of leaves and secondary vascular tissues in Quercus pubescens

An increased frequency of fire events on the Slovenian Karst is in line with future climate-change scenarios for drought-prone environments worldwide. It is therefore of the utmost importance to better understand tree-fire-climate interactions for predicting the impact of changing environment on tree functioning. To this purpose, we studied the post-fire effects on leaf development, leaf carbon isotope composition (%13C), radial growth patterns and the xylem and phloem anatomy in undamaged (H-trees) and fire-damaged trees (F-trees) of Q. pubescens with good re-sprouting ability in spring 2017, the growing season after a rangeland fire in August 2016. We found that the fully developed canopy of F-trees reached only half of the LAI values measured in H-trees. Throughout the season, F-trees were characterised by higher water potential and stomatal conductivity and achieved higher photosynthetic rates compared to unburnt H-trees. The foliage of F-trees had more negative %13C values than those of H-trees. This reflects that F-trees less frequently meet stomatal limitations due to reduced transpirational area and more favourable leaf-to-root ratio. In addition, the growth of leaves in F-trees relied more on the recent photosynthates than on reserves due to the fire disturbed starch accumulation in the previous season. Cambial production stopped 3 weeks later in F-trees, resulting in 60% and 22% wider xylem and phloem increments, respectively. A novel approach by including phloem anatomy in the analyses revealed that fire caused changes in conduit dimensions in the early phloem but not in the earlywood. However, premature formation of the tyloses in the earlywood vessels of the youngest two xylem increments in F-trees implies that xylem hydraulic integrity was also affected by heat. Analyses of secondary tissues showed that although xylem and phloem tissues are interlinked changes in their transport systems due to heat damage are not necessarily coordinated

Effects of Specific Parameters on Simulations of Energy Use and Air Temperatures in Offices Equipped with Radiant Heating/Cooling Panels

When creating a simulation model to assess the performance of buildings, there is usually a lack of feedback information. Only in the case of measurements of a real building is a direct comparison of the measured values and simulated results possible. Parameter data related to users&rsquo; behavior or other events can also be obtained. Their evaluated frequency, magnitude and duration, along with boundary conditions, are crucial for the results. It is clear that none of them can be predicted very accurately. Most of them, however, are needed for computer modeling. In this paper we analyzed the well-defined TRNSYS simulation model of offices equipped with radiant ceiling panels for heating and cooling. The model was based on real case offices and was validated based on measurements for 1 year. The analysis included simulations in order to define what effect the parameters related mainly to users have on the energy use and the indoor air temperatures. The study confirmed that specific human activities influence the annual energy use to a relatively small degree and that their effects often counteract. It also confirmed the even more important fact that although small, these activities can influence the thermal comfort of users. It is believed that despite the fact that this research was based on an analysis of offices equipped with radiant ceiling panels, most of the results could be applied generally

Delirprävention – das Basler Modell

Based on actual evidence, about one third of deliria can be prevented. Hereby, preventive measures can address both delirium predisposing as well as delirium precipitating factors