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Abstract Recently it has been proposed that methane might be produced by all living organisms via a mechanism driven by reactive oxygen species that arise through the metabolic activity of cells. Here, we summarise details of this novel reaction pathway and discuss its potential significance for clinical and health sciences. In particular, we highlight the role of oxidative stress in cellular methane formation. As several recent studies also demonstrated the anti‐inflammatory potential for exogenous methane‐based approaches in mammalians, this article addresses the intriguing question if ROS‐driven methane formation has a general physiological role and associated diagnostic potential.

Post-traumatic stress disorder (PTSD) is a debilitating illness characterized by dysfunction in the medial prefrontal cortex (mPFC). Although both pharmacological and cognitive behavioral interventions have shown some promise at alleviating symptoms, high attrition and persistence of treatment-resistant symptoms pose significant challenges that remain unresolved. Specifically, prolonged exposure therapy, a gold standard intervention to treat PTSD, has high dropout rates resulting in many patients receiving less than a fully effective course of treatment. Administering pharmacological treatments together with behavioral psychotherapies like prolonged exposure may offer an important avenue for enhancing therapeutic efficacy sooner, thus reducing the duration of treatment and mitigating the impact of attrition. In this study, using extinction learning as a rat model of exposure therapy, we hypothesized that administering ketamine as an adjunct treatment together with extinction will enhance the efficacy of extinction in reversing stress-induced deficits in set shifting, a measure of cognitive flexibility. Results showed that combining a sub-effective dose of ketamine with a shortened, sub-effective extinction protocol fully reversed stress-induced cognitive set-shifting deficits in both male and female rats. These effects may be due to shared molecular mechanisms between extinction and ketamine, such as increased neuronal plasticity in common circuitry (e.g., hippocampus-mPFC), or increased BDNF signaling. This work suggests that fast-acting drugs, such as ketamine, can be effectively used in combination with behavioral interventions to reduce treatment duration and potentially mitigate the impact of attrition. Future work is needed to delineate other pharmacotherapies that may complement the effects of extinction via shared or independent mechanisms.

Purpose: In medial open-wedge high tibial osteotomy (HTO) hinge axis and osteotomy plane influence the resulting anatomy, but accurate angular quantifications using 3D-planning-simulations are lacking. The objectives of this study were developing a standardized and validated 3D-planning method of an HTO and to perform several simulated realignments to explain unintended anatomy changes. Methods: The cutting direction of the main osteotomy was defined parallel to the medial tibial slope and the hinge axis 1.5 cm distal to the lateral plateau. For interobserver testing, this 3D planning was performed on 13 digital models of human tibiae by two observers. In addition, four different hinge axis positions and five differently inclined osteotomy planes each were simulated. The osteotomy direction ranged from medial 0°–30° anteromedial, while the tilt of the osteotomy plane compared to the tibial plateau was −10° to +10°. All anatomic angular changes were calculated using 3D analysis. Results: Multiple HTO plannings by two medical investigators using standardized procedures showed only minimal differences. In the 3D-simulation, each 10° rotation of the hinge axis resulted in a 1.7° significant increase in slope. Tilting the osteotomy plane by 10° resulted in significant torsional changes of 2°, in addition to minor but significant changes in the medial proximal tibial angle (MPTA). Conclusion: Standardized 3D-planning of the HTO can be performed with high reliability using two-observer planning. 3D-simulations suggest that control of the osteotomy plane is highly relevant to avoid unintended changes in the resulting anatomy, but this can be a helpful tool to modify specific angles in different pathologies in the HTO.

Blow flies (Calliphoridae) are used in forensic entomology studies, because their life cycle help to determine the minimum post-mortem interval (PMImin). Paralucilia fulvinota (Bigot, 1877) is a forensic indicator and is reported from human corpses in the Amazonian region. Our objectives were to describe the development time of P. fulvinota under natural conditions at Ducke Reserve (Amazonas), provide the accumulated degree-days (ADD) and accumulated degree-hour (ADH), body length and mass of larvae, and the sex ratio of adults. Pregnant females were sampled on a swine carcass. From hatched eggs, we reared stock colonies and observed the immature development under uncontrolled conditions of temperature (24.5 ± 1.3 ºC), rainfall (188 ± 11.5 mm³) and humidity (84 ± 3.9%). Development time was 11 days. Adult sex ratio was 0.86. Larvae gained mass at constant rates through the instars but grew less in length from the second to the third instar. Accordingly, P. fulvinota grows more in mass than in length when in larval stage. The period from first stadium to adults lasted 159.5 degree-days and 3828 degree-hours. Our results may be helpful in providing a more accurate estimate of PMImin on corpses encountered in forested areas of the Amazon region.

Abstract Mechanotransduction via yes-associated protein (YAP) is a central mechanism for decision-making in mouse embryonic stem cells (mESCs). Nuclear localization of YAP is tightly connected to pluripotency and increases the cell division rate (CDR). How the geometry of the extracellular environment influences mechanotransduction, thereby YAP localization, and decision-making of single isolated mESCs is largely unknown. To investigate this relation, we produced well-defined 2D and 2.5D microenvironments and monitored CDR and subcellular YAP localization in single mESCs hence excluding cell–cell interactions. By systematically varying size and shape of the 2D and 2.5D substrates we observed that the geometry of the growth environment affects the CDR. Whereas CDR increases with increasing adhesive area in 2D, CDR is highest in small 2.5D micro-wells. Here, mESCs attach to all four walls and exhibit a cross-shaped cell and nuclear morphology. This observation indicates that changes in cell shape are linked to a high CDR. Inhibition of actomyosin activity abrogate these effects. Correspondingly, nuclear YAP localization decreases in inhibitor treated cells, suggesting a relation between cell shape, intracellular forces, and cell division rate. The simplicity of our system guarantees high standardization and reproducibility for monitoring stem cell reactions and allows addressing a variety of fundamental biological questions on a single cell level.

Purpose: Functional imaging is often used to try to elucidate the pathophysiological mechanism of tinnitus. Residual inhibition, the temporary suppression of tinnitus after application of a masking noise, could be an interesting technique to modulate tinnitus perception in functional imaging paradigms. The purposes of this study were to primarily assess reproducibility of the (partial) positive residual inhibition response duration in patients with tinnitus and to explore its utility in experimental designs. Method: Patients with tinnitus exhibiting a (partial) positive residual inhibition response or tinnitus reduction after a 1-min white noise presentation were selected from a broad consulting tinnitus population. In 27 patients, this response was tested 4 times: twice during initial testing and twice during a retest of the psychoacoustic tinnitus measures, 4-8 weeks after initial consultation. In 17 patients with stable residual inhibition responses, reproducibility of response duration, the duration of tinnitus reduction up to pretesting state, was analyzed. Results: Initial testing showed a residual inhibition duration of 29.5 s on average. Test-retest reproducibility of response duration was shown to be reliable with an ICC(3, 4) of 0.871 (95% CI [0.733, 0.948]) and a standard error of measurement of 6.64 s. Conclusions: This study indicates the good test-retest reproducibility of residual inhibition duration in our subset of 17 patients with stable (partial) positive residual inhibition. Residual inhibition is, therefore, a technique that can potentially be used for temporary tinnitus manipulation in experimental paradigms to unravel tinnitus pathophysiology.

Based on their drug delivery properties and activity against tumors, we combined PAMAM dendrimers with various platinum(IV) complexes in order to provide an improved approach of anticancer treatment. Platinum(IV) complexes were linked to terminal NH2 moieties of PAMAM dendrimers of generation 2 (G2) and 4 (G4) via amide bonds. Conjugates were characterized by 1H and 195Pt NMR spectroscopy, ICP-MS and in representative cases by pseudo-2D diffusion-ordered NMR spectroscopy. Additionally, the reduction behavior of conjugates in comparison to corresponding platinum(IV) complexes was investigated, showing a faster reduction of conjugates. Cytotoxicity was evaluated via the MTT assay in human cell lines (A549, CH1/PA-1, SW480), revealing IC50 values in the low micromolar to high picomolar range. The synergistic combination of PAMAM dendrimers and platinum(IV) complexes resulted in up to 200 times increased cytotoxic activity of conjugates in consideration of the loaded platinum(IV) units compared to their platinum(IV) counterparts. The lowest IC50 value of 780 ± 260 pM in the CH1/PA-1 cancer cell line was detected for an oxaliplatin-based G4 PAMAM dendrimer conjugate. Finally, in vivo experiments of a cisplatin-based G4 PAMAM dendrimer conjugate were performed based on the best toxicological profile. A maximum tumor growth inhibition effect of 65.6% compared to 47.6% for cisplatin was observed as well as a trend of prolonged animal survival.

A new class of anticancer prodrugs was designed by combining the cytotoxicity of platinum(IV) complexes and the drug carrier properties of glycol chitosan polymers: Unsymmetrically carboxylated platinum(IV) analogues of cisplatin, carboplatin and oxaliplatin, namely (OC-6-44)-acetatodiammine(3-carboxypropanoato)dichloridoplatinum(IV), (OC-6-44)-acetaodiammine(3-carboxypropanoato)(cyclobutane-1,1-dicarboxylato)platinum(IV) and (OC-6-44)-acetato(3-carboxypropanoato)(1R,2R-cyclohexane-1,2-diamine)oxalatoplatinum(IV) were synthesised and conjugated via amide bonding to degraded glycol chitosan (dGC) polymers with different chain lengths (5, 10, 18 kDa). The 15 conjugates were investigated with 1H and 195Pt NMR spectroscopy, and average amounts of platinum(IV) units per dGC polymer molecule with ICP-MS, revealing a range of 1.3–22.8 platinum(IV) units per dGC molecule. Cytotoxicity was tested with MTT assays in the cancer cell lines A549, CH1/PA-1, SW480 (human) and 4T1 (murine). IC50 values in the low micromolar to nanomolar range were obtained, and higher antiproliferative activity (up to 72 times) was detected with dGC-platinum(IV) conjugates in comparison to platinum(IV) counterparts. The highest cytotoxicity (IC50 of 0.036 ± 0.005 µM) was determined in CH1/PA-1 ovarian teratocarcinoma cells with a cisplatin(IV)–dGC conjugate, which is hence 33 times more potent than the corresponding platinum(IV) complex and twice more potent than cisplatin. Biodistribution studies of an oxaliplatin(IV)–dGC conjugate in non-tumour-bearing Balb/C mice showed an increased accumulation in the lung compared to the unloaded oxaliplatin(IV) analogue, arguing for further activity studies.

The RASopathies are a group of disorders due to variations of genes associated with the Ras/MAPK pathway. Some of the RASopathies include neurofibromatosis type 1 (NF1), Noonan syndrome, Noonan syndrome with multiple lentigines, cardiofaciocutaneous (CFC) syndrome, Costello syndrome, Legius syndrome, and capillary malformation-arteriovenous malformation (CM-AVM) syndrome. In combination, the RASopathies are a frequent group of genetic disorders. This report summarizes the proceedings of the 4th International Symposium on Genetic Disorders of the Ras/MAPK pathway and highlights gaps in the field. © 2016 Wiley Periodicals, Inc.

Previous research suggests that the far-reaching impacts of hurricanes include the academic performance of students. In an examination of such impacts, we found a trend toward self-perceived decline in some performance indicators relative to students at peer universities. However, few longitudinal impacts were found, perhaps because of the sense of community offered by the living-learning program. These results may inform administrators and faculty of areas for emphasis in mitigating future impacts.

Methane (CH4) is the most abundant organic compound in the atmosphere and is emitted from many biotic and abiotic sources. Recent studies have shown that CH4 production occurs under aerobic conditions in eukaryotes, such as plants, animals, algae, and saprotrophic fungi. Saprotrophic fungi play an important role in nutrient recycling in terrestrial ecosystems via the decomposition of plant litter. Although CH4 production by saprotrophic fungi has been reported, no data on the stable carbon isotope values of the emitted CH4 (δ13C-CH4 values) are currently available. In this study, we measured the δ13C values of CH4 and carbon dioxide (δ13C-CO2 values) emitted by two saprotrophic fungi, Pleurotus sapidus (oyster mushroom) and Laetiporus sulphureus (sulphur shelf), cultivated on three different substrates, pine wood (Pinus sylvestris), grass (mixture of Lolium perenne, Poa pratensis, and Festuca rubra), and corn (Zea mays), which reflect both C3 and C4 plants with distinguished bulk δ13C values. Applying Keeling plots, we found that the δ13C source values of CH4 emitted from fungi cover a wide range from −40 to −69 mUr depending on the growth substrate and fungal species. Whilst little apparent carbon isotopic fractionation (in the range from −0.3 to 4.6 mUr) was calculated for the δ13C values of CO2 released from P. sapidus and L. sulphureus relative to the bulk δ13C values of the growth substrates, much larger carbon isotopic fractionations (ranging from −22 to −42 mUr) were observed for the formation of CH4. Although the two fungal species showed similar δ13CH4 source values when grown on pine wood, δ13CH4 source values differed substantially between the two fungal species when they were grown on grass or corn. We found that the source values of δ13CH4 emitted by saprotrophic fungi are highly dependent on the fungal species and the metabolized substrate. The source values of δ13CH4 cover a broad range and overlap with values reported for methanogenic archaea, the thermogenic degradation of organic matter, and other eukaryotes.

Abstract Background Starting in December 2019, the current pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) confronts the world with an unprecedented challenge. With no vaccine or drug being currently available to control the pandemic spread, prevention and PCR (Polymerase chain reaction) testing becomes a crucial pillar of medical systems. Aim of the present study was to report on the first results of the measures taken in a large German Department of Radiation Oncology, including PCR testing of asymptomatic cancer patients. Methods Pandemic-adapted hygiene regulations and prevention measures for patients and staff were implemented. A visiting ban on both wards was implemented from the beginning and medical staff and patients were required to wear face masks at all times. The waiting rooms were rearranged to ensure distance between patients of at least 1.5 m. Clinical follow up was mainly done by telephone and all patients had to complete a questionnaire regarding symptoms and contacts with COVID-19 patients before entering our department. Educational documents were created for patients to raise awareness of symptoms and avoidance strategies for interactions with other people. Indications for therapy and fractionation schemes were adapted when possible. In a subsequent step, all new asymptomatic patients were tested via nasopharyngeal swab at our screening station shortly before their simulation CT. Results All these measures and implementations have been well accepted semiquantitatively measured by the consent received from patients and staff. Regarding the PCR testing, only 1 out of 139 asymptomatic patients of our cohort so far tested positive for SARS-CoV-2, reflecting a prevalence of 0.72% in this cancer patient population. Up to this point no staff members was tested positive. The start of the treatment for the PCR-positive patient was deferred for 2 weeks. Conclusion Due to the pandemic-adapted implementations, our department seems well prepared during this crisis. The initial screening helps to identify asymptomatic COVID-19 patients in order to protect other patients and our staff from infection and the observed PCR prevalence is in line with comparable studies. A regular PCR testing (e.g. twice a week) of all patients and staff would in principle be desirable but is limited due to testing capacities at present.

Abstract Background Reconstruction of metaphyseal fractures represents a clinical challenge for orthopedic surgeons. Especially in osteoporotic bone, these fractures are frequently accompanied by osseous substance defects. In order to ensure rapid mobilization of patients, high stability requirements must be met by osteosynthesis. Various bone graft materials have been introduced in the past, such as autologous bone or exogenous bone substitute materials. These are used as bone void fillers or as augmentation techniques to ensure safe fixation of osteosynthesis. New calcium phosphate-based bone void-filling materials could be a promising alternative to autologous bone or to the currently and widely used polymethylmethacrylate (PMMA)-based cement. The aim of this study was to evaluate a novel paste-like bone void filler in vivo and in vitro with regard to biocompatibility and osteoconductivity. Methods In addition to in vitro testing of cell compatibility using pre-osteoblasts (MC3T3-E1), 35 Wistar rats were treated in vivo with implantation of various material mixtures based on calcium phosphate and aluminum oxide reinforcement in a metaphyseal drill hole defect. After 4 weeks, an examination by micro-computed tomography (μCT) and histology was performed. Results The in vitro analysis showed good biocompatibility with a high cell survival of osteoblasts. In the in vivo experiments, a significantly higher bone ingrowth compared to the empty defect was shown by μCT and histological analysis. Here, the group receiving material reinforced with aluminum oxide (Al2O3) showed a bone volume/tissue volume (BV/TV) of 89.19% compared to a BV/TV of 83.14% for the empty defect (p = 0.0013). In the group treated with a polysaccharide matrix, no increase in BV/TV was observed given a mean ratio of 80.14%. Scoring of histological sections did not reveal a significant difference between CaP and CaP that was substituted with Al2O3. Conclusion The results of this study show an encouraging first step towards the development of new pasty, bone void-filling materials. We demonstrated that a new paste-like bone-filling material, based on calcium phosphate granulates and aluminum oxide to provide strength, exhibits good biocompatibility and osteoconductivity. Further biomechanical test in an osteoporotic animal model will have to be performed, to prove feasibility in metaphyseal defects.

FCH domain only 1 (FCHO1) is a key molecule involved in clathrin-mediated endocytosis (CME). Here, the authors report homozygous FCHO1 mutations in individuals with variable T and B cell lymphopenia, which are associated with loss-of-function of FCHO1 and impaired formation of clathrin-coated pits in T cells.

Winter cover crops have been shown to limit erosion and nutrient runoff from agricultural land. To promote their usage, the Maryland Department of Agriculture (MDA) subsidizes farmers who plant cover crops. Conventional verification of cover crop planting and analysis of subsequent crop performance requires on-the-ground fieldwork, which is costly and labor intensive. In partnership with the MDA, NASA's DEVELOP program utilized imagery from Landsat 5, Landsat 8, and the European Space Agency’s Sentinel-2 to create a decision support tool for satellite-based monitoring of cover crop performance throughout Maryland. Our teams created CCROP, an interactive graphical user interface, in Google Earth Engine which analyzes satellite imagery to calculate the normalized difference vegetation index (NDVI) of fields across the state. Linear regression models were applied to convert NDVI to estimates of crop biomass and percent green ground cover, with measure of fit (R2) values ranging from 0.4 to 0.7. These crop metrics were implemented into an interactive filtering tool within CCROP which allows users to examine cover crop performance based on a variety of growing parameters. CCROP also includes a time series analysis routine for examining the progression of NDVI throughout the spring to help determine farmer-induced termination dates of cover crops. With this decision support tool, the MDA can analyze the effectiveness of cover crops throughout the state with reduced need to manually spot-check enrolled production fields, and can identify variables influencing overall cover crop performance to optimize implementation of their winter cover crop program via adaptive management approaches.

Cigarette smoke (CS) causes adverse health effects and, for smoker who do not quit, modified risk tobacco products (MRTPs) can be an alternative to reduce the risk of developing smoking-related diseases. Standard toxicological endpoints can lack sensitivity, with systems toxicology approaches yielding broader insights into toxicological mechanisms. In a 6-month systems toxicology study on ApoE−/− mice, we conducted an integrative multi-omics analysis to assess the effects of aerosols from the Carbon Heated Tobacco Product (CHTP) 1.2 and Tobacco Heating System (THS) 2.2—a potential and a candidate MRTP based on the heat-not-burn (HnB) principle—compared with CS at matched nicotine concentrations. Molecular exposure effects in the lungs were measured by mRNA/microRNA transcriptomics, proteomics, metabolomics, and lipidomics. Integrative data analysis included Multi-Omics Factor Analysis and multi-modality functional network interpretation. Across all five data modalities, CS exposure was associated with an increased inflammatory and oxidative stress response, and lipid/surfactant alterations. Upon HnB aerosol exposure these effects were much more limited or absent, with reversal of CS-induced effects upon cessation and switching to CHTP 1.2. Functional network analysis revealed CS-induced complex immunoregulatory interactions across the investigated molecular layers (e.g., itaconate, quinolinate, and miR-146) and highlighted the engagement of the heme–Hmox–bilirubin oxidative stress axis by CS. This work exemplifies how multi-omics approaches can be leveraged within systems toxicology studies and the generated multi-omics data set can facilitate the development of analysis methods and can yield further insights into the effects of toxicological exposures on the lung of mice.

Classical antiviral restriction factors promote cellular immunity by their ability to interfere with virus replication and induction of their expression by proinflammatory cytokines such as interferons. The serine incorporator proteins SERINC3 and SERINC5 potently reduce the infectivity of HIV-1 particles when overexpressed, and RNA interference or knockout approaches in T cells have indicated antiviral activity also of the endogenous proteins. Due to lack of reagents for detection of endogenous SERINC proteins, it is still unclear whether SERINC3/5 are expressed to functionally relevant levels in different primary target cells of HIV infection and how the expression levels of these innate immunity factors are regulated. In the current study, analysis of SERINC3/5 mRNA steady-state levels in primary lymphoid and monocyte-derived cells revealed selective induction of their expression upon differentiation of myeloid cells. Contrary to classical antiviral restriction factors, various antiviral α-interferon subtypes and proinflammatory interleukins had no effect on SERINC levels, which were also not dysregulated in CD4+ T cells and monocytes isolated from patients with chronic HIV-1 infection. Notably, HIV-1 particles produced by terminally differentiated monocyte-derived macrophages with high SERINC5 expression, but not by low-expressing monocytes, showed a Nef-dependent infectivity defect. Overall, these findings suggest endogenous expression of SERINC5 to antivirally active levels in macrophages. Our results classify SERINC5 as an unconventional HIV-1 restriction factor whose expression is specifically induced upon differentiation of cells towards the myeloid lineage.

In the agricultural industry, the conservation of wheat grains in silos after stocking is a hard task. This is because when stocking a large crop, hot aeration from the bottom of the silo is not sufficient to preserve the crop and avoid losses due to the manifestation of moisture between grains and grains-wall. As a concern, engineers, as well as inspectors, challenged day-to-day issues whenever packing such granular material. A screw could be a good solution to improve the dryness of the particles and mix the bed material in case of stocking different types of grains. Mainly, the complexity does emanate from the model used to predict accurately the mixing performance. Emerging generation of simulation codes using the discrete element method (e.g., EDEM®, LIGGGHTS, etc.) have brought an intact solution to the problem of particulate flow and particles inhomogeneity. The present work has sought to develop predictive models for average mixing index and particles velocity distribution assessments in mixing of wheat granules in a small-scaled hopper base screw mixer. A screw-mated-horizontal-ploughs has been utilized to improve the agitation of grains rather than a simple screw. 1, 2 and 4 mated ploughs to the screw axle has been used, also the tilt angle of the 4 mated ploughs case has been varied as follows; 20°, 45° and 70° then evaluated. Next, findings regarding bed homogeneity are calculated using the Lacey method [1] for quantitative analysis and visual observations have been read out for qualitative assessment. Particles inhomogeneity is at its minimum when more ploughs mated to the screw were used.

BackgroundAssessment of the physical frailty of older patients is of great importance in many medical disciplines to be able to implement individualized therapies. For physical tests, time is usually used as the only objective measure. To record other objective factors, modern wearables offer great potential for generating valid data and integrating the data into medical decision-making. ObjectiveThe aim of this study was to compare the predictive value of insole data, which were collected during the Timed-Up-and-Go (TUG) test, to the benchmark standard questionnaire for sarcopenia (SARC-F: strength, assistance with walking, rising from a chair, climbing stairs, and falls) and physical assessment (TUG test) for evaluating physical frailty, defined by the Short Physical Performance Battery (SPPB), using machine learning algorithms. MethodsThis cross-sectional study included patients aged >60 years with independent ambulation and no mental or neurological impairment. A comprehensive set of parameters associated with physical frailty were assessed, including body composition, questionnaires (European Quality of Life 5-dimension [EQ 5D 5L], SARC-F), and physical performance tests (SPPB, TUG), along with digital sensor insole gait parameters collected during the TUG test. Physical frailty was defined as an SPPB score≤8. Advanced statistics, including random forest (RF) feature selection and machine learning algorithms (K-nearest neighbor [KNN] and RF) were used to compare the diagnostic value of these parameters to identify patients with physical frailty. ResultsClassified by the SPPB, 23 of the 57 eligible patients were defined as having physical frailty. Several gait parameters were significantly different between the two groups (with and without physical frailty). The area under the receiver operating characteristic curve (AUROC) of the TUG test was superior to that of the SARC-F (0.862 vs 0.639). The recursive feature elimination algorithm identified 9 parameters, 8 of which were digital insole gait parameters. Both the KNN and RF algorithms trained with these parameters resulted in excellent results (AUROC of 0.801 and 0.919, respectively). ConclusionsA gait analysis based on machine learning algorithms using sensor soles is superior to the SARC-F and the TUG test to identify physical frailty in orthogeriatric patients.

Summary: Although some COVID-19 patients maintain SARS-CoV-2-specific serum immunoglobulin G (IgG) for more than 6 months postinfection, others eventually lose IgG levels. We assessed the persistence of SARS-CoV-2-specific B cells in 17 patients, 5 of whom had lost specific IgGs after 5–8 months. Differentiation of blood-derived B cells in vitro revealed persistent SARS-CoV-2-specific IgG B cells in all patients, whereas IgA B cells were maintained in 11. Antibodies derived from cultured B cells blocked binding of viral receptor-binding domain (RBD) to the cellular receptor ACE-2, had neutralizing activity to authentic virus, and recognized the RBD of the variant of concern Alpha similarly to the wild type, whereas reactivity to Beta and Gamma were decreased. Thus, differentiation of memory B cells could be more sensitive for detecting previous infection than measuring serum antibodies. Understanding the persistence of SARS-CoV-2-specific B cells even in the absence of specific serum IgG will help to promote long-term immunity.

For a variety of cancer types, platinum compounds are still among the best treatment options. However, their application is limited by side effects and drug resistance. Consequently, multi-targeted platinum(IV) prodrugs that target specific traits of the malignant tissue are interesting new candidates. Recently, cisPt(PhB)2 was synthesized which, upon reduction in the malignant tissue, releases phenylbutyrate (PhB), a metabolically active fatty acid analog, in addition to cisplatin. In this study, we in-depth investigated the anticancer properties of this new complex in cell culture and in mouse allograft experiments. CisPt(PhB)2 showed a distinctly improved anticancer activity compared to cisplatin as well as to PhB alone and was able to overcome various frequently occurring drug resistance mechanisms. Furthermore, we observed that differences in the cellular fatty acid metabolism and mitochondrial activity distinctly impacted the drug’s mode of action. Subsequent analyses revealed that “Warburg-like” cells, which are characterized by deficient mitochondrial function and fatty acid catabolism, are less capable of coping with cisPt(PhB)2 leading to rapid induction of a non-apoptotic form of cell death. Summarizing, cisPt(PhB)2 is a new orally applicable platinum(IV) prodrug with promising activity especially against cisplatin-resistant cancer cells with “Warburg-like” properties.

Cancer ablation therapies aim to be efficient while minimizing damage to healthy tissues. Nanosecond pulsed electric field (nsPEF) is a promising ablation modality because of its selectivity against certain cell types and reduced neuromuscular effects. We compared cell killing efficiency by PEF (100 pulses, 200 ns–10 µs duration, 10 Hz) in a panel of human esophageal cells (normal and pre-malignant epithelial and smooth muscle). Normal epithelial cells were less sensitive than the pre-malignant ones to unipolar PEF (15–20% higher LD50, p < 0.05). Smooth muscle cells (SMC) oriented randomly in the electric field were more sensitive, with 30–40% lower LD50 (p < 0.01). Trains of ten, 300-ns pulses at 10 kV/cm caused twofold weaker electroporative uptake of YO-PRO-1 dye in normal epithelial cells than in either pre-malignant cells or in SMC oriented perpendicularly to the field. Aligning SMC with the field reduced the dye uptake fourfold, along with a twofold reduction in Ca2+ transients. A 300-ns pulse induced a twofold smaller transmembrane potential in cells aligned with the field, making them less vulnerable to electroporation. We infer that damage to SMC from nsPEF ablation of esophageal malignancies can be minimized by applying the electric field parallel to the predominant SMC orientation.

Lead (Pb) is a highly toxic heavy metal that has deleterious effects on the central nervous system. This study aimed to investigate the effects of salinomycin (Sal) and deferiprone (DFP) on brain morphology and on the content of some essential elements in Pb-exposed mice. Adult male Institute of Cancer Research (ICR) mice were exposed to a daily dose of 80 mg/kg body weight ( b.w.) Pb(II) nitrate for 14 days and subsequently treated with Sal (16 mg/kg b.w.) or DFP (19 mg/kg b.w.) for another 14 days. At the end of the experimental protocol, the brains were processed for histological and inductively coupled plasma mass spectrometry (ICP-MS) analyses. Pb exposure resulted in a 50-fold increase in Pb concentration, compared with controls. Magnesium (Mg) and phosphorus (P) were also significantly increased by 22.22% and 17.92%, respectively. The histological analysis of Pb-exposed mice revealed brain pathological changes with features of neuronal necrosis. Brain Pb level remained significantly elevated in Sal- and DFP-administered groups (37-fold and 50-fold, respectively), compared with untreated controls. Treatment with Sal significantly reduced Mg and P concentrations by 22.56% and 18.38%, respectively, compared with the Pb-exposed group. Administration of Sal and DFP ameliorated brain injury in Pb-exposed mice and improved histological features. The results suggest the potential application of Sal and DFP for treatment of Pb-induced neurotoxicity.

Giant exoplanets have been directly imaged over orders of magnitude of orbital separations, prompting theoretical and observational investigations of their formation pathways. In this paper, we present new VLTI/GRAVITY astrometric data of HIP 65426 b, a cold, giant exoplanet which is a particular challenge for most formation theories at a projected separation of 92 au from its primary. Leveraging GRAVITY’s astrometric precision, we present an updated eccentricity posterior that disfavors large eccentricities. The eccentricity posterior is still prior dependent, and we extensively interpret and discuss the limits of the posterior constraints presented here. We also perform updated spectral comparisons with self-consistent forward-modeled spectra, finding a best-fit ExoREM model with solar metallicity and C/O = 0.6. An important caveat is that it is difficult to estimate robust errors on these values, which are subject to interpolation errors as well as potentially missing model physics. Taken together, the orbital and atmospheric constraints paint a preliminary picture of formation inconsistent with scattering after disk dispersal. Further work is needed to validate this interpretation. Analysis code used to perform this work is available on GitHub: https://github.com/sblunt/hip65426 .

Tension remains between the observed and modeled properties of substellar objects, but objects in binary orbits, with known dynamical masses, can provide a way forward. HD 72946 B is a recently imaged brown dwarf companion to a nearby, solar-type star. We achieve ∼100 μ as relative astrometry of HD 72946 B in the K band using VLTI/GRAVITY, unprecedented for a benchmark brown dwarf. We fit an ensemble of measurements of the orbit using orbitize! and derive a strong dynamical mass constraint M _B = 69.5 ± 0.5 M _Jup assuming a strong prior on the host star mass M _A = 0.97 ± 0.01 M _⊙ from an updated stellar analysis. We fit the spectrum of the companion to a grid of self-consistent BT-Settl-CIFIST model atmospheres, and perform atmospheric retrievals using petitRADTRANS . A dynamical mass prior only marginally influences the sampled distribution of effective temperature, but has a large influence on the surface gravity and radius, as expected. The dynamical mass alone does not strongly influence retrieved pressure–temperature or cloud parameters within our current retrieval setup. Independently of the cloud prescription and prior assumptions, we find agreement within ±2 σ between the C/O of the host (0.52 ± 0.05) and brown dwarf (0.43–0.63), as expected from a molecular cloud collapse formation scenario, but our retrieved metallicities are implausibly high (0.6–0.8) in light of the excellent agreement of the data with the solar-abundance model grid. Future work on our retrieval framework will seek to resolve this tension. Additional study of low surface gravity objects is necessary to assess the influence of a dynamical mass prior on atmospheric analysis.

The recent discovery of enormous Ly α nebulae (ELANe), characterized by physical extents >200 kpc and Ly α luminosities >10 ^44 erg s ^−1 , provides a unique opportunity to study the intergalactic medium and circumgalactic medium in distant galaxies. Many existing ELANe detections are associated with local overdensities of active galactic nuclei (AGN). We have initiated a search for ELANe around regions containing pairs of quasi-stellar objects (QSOs) using the Palomar Cosmic Web Imager. The first study of this search, Cai et al., presented results of ELAN0101+0201, which was associated with a QSO pair at z = 2.45. In this study, all targets residing in QSO pair environments analyzed have Ly α detections, but only one of the four targets meets the classification criteria of an ELANe associated with a QSO pair region ( z ∼ 2.87). The other three sample detections of Ly α nebulae do not meet the size and luminosity criteria to be classified as ELANe. We find kinematic evidence that the ELANe J1613 is possibly powered mostly by AGN outflows. The analysis of circularly averaged surface brightness profiles of emission from the Ly α regions shows that the Ly α emission around z ∼ 2 QSO pairs is consistent with emission around individual QSOs at z ∼ 2, which is fainter than that around z ∼ 3 QSOs. A larger sample of Ly α at z ∼ 2 will be needed to determine if there is evidence of redshift evolution when compared to nebular emissions at z ∼ 3 from other studies.

Cancer chemotherapeutics usually have serious side effects. Targeting the special properties of cancer and activation of the anticancer drug in the tumor microenvironment in situ may decrease the intensity of the side effects and improve the efficacy of therapy. In this study, half-sandwich Rh complexes are introduced, which may be activated at the acidic, extracellular pH of the tumor tissue. The synthesis and aqueous stability of mixed-ligand complexes with a general formula of [Rh(η5-Cp*)(N,N/O)(N)]2+/+ are reported, where (N,N/O) indicates bidentate 8-quinolate, ethylenediamine and 1,10-phenanthroline and (N) represents the releasable monodentate ligand with a nitrogen donor atom. UV-visible spectrophotometry, 1H NMR, and pH-potentiometry were used to determine the protonation constants of the monodentate ligands, the proton dissociation constants of the coordinated water molecules in the aqua complexes, and the formation constants of the mixed-ligand complexes. The obtained data were compared to those of the analogous Ru(η6-p-cymene) complexes. The developed mixed-ligand complexes were tested in drug-sensitive and resistant colon cancer cell lines (Colo205 and Colo320, respectively) and in four bacterial strains (Gram-positive and Gram-negative, drug-sensitive, and resistant) at different pH values (5–8). The mixed-ligand complexes with 1-methylimidazole displayed sufficient stability at pH 7.4, and their activation was found in cancer cells with decreasing pH; moreover, the mixed-ligand complexes demonstrated antimicrobial activity in Gram-positive and Gram-negative bacteria, including the resistant MRSA strain. This study proved the viability of incorporating releasable monodentate ligands into mixed-ligand half-sandwich complexes, which is supported by the biological assays.

A growing global emission of engineered nanoparticles (ENPs) into the aquatic environment has become an emerging safety concern that requires methods capable of identifying the occurrence and possibly determining the amounts of ENPs. In this study, we employed sector-field inductively coupled mass spectrometry to assess the presence of ENPs in coastal seawater samples collected from the Black Sea in regions suffering different anthropogenic impacts. Ultrafiltration through commercial 3 kDa membrane filters was shown to be feasible to separate the ENPs from the bulk seawater, and the subsequent ultrasound-mediated acidic dissolution makes the metals constituting the ENPs amenable to analysis. This procedure allowed the ENPs bearing Cu, Zn, V, Mo, and Sn to be for the first time quantitated in seashore surface water, their concentration ranging from 0.1 to 1.0 μg L−1 (as metal) and related to the presence of industry and/or urban stress. While these levels are decreased by natural dilution and possible sedimentation, the monitored ENPs remain measurable at a distance of 2 km from the coast. This can be attributed not only to local emission sources but also to some natural backgrounds.

Coccidioides immitis and Coccidioides posadasii are soil-dwelling fungi of arid regions in North and South America that are responsible for Valley fever (coccidioidomycosis). Forty percent of patients with Valley fever exhibit symptoms ranging from mild, self-limiting respiratory infections to severe, life-threatening pneumonia that requires treatment. Misdiagnosis as bacterial pneumonia commonly occurs in symptomatic Valley fever cases, resulting in inappropriate treatment with antibiotics, increased medical costs, and delay in diagnosis. In this proof-of-concept study, we explored the feasibility of developing breath-based diagnostics for Valley fever using a murine lung infection model. To investigate potential volatile biomarkers of Valley fever that arise from host–pathogen interactions, we infected C57BL/6J mice with C. immitis RS (n = 6), C. posadasii Silveira (n = 6), or phosphate-buffered saline (n = 4) via intranasal inoculation. We measured fungal dissemination and collected bronchoalveolar lavage fluid (BALF) for cytokine profiling and for untargeted volatile metabolomics via solid-phase microextraction (SPME) and two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC×GC-TOFMS). We identified 36 volatile organic compounds (VOCs) that were significantly correlated (p < 0.05) with cytokine abundance. These 36 VOCs clustered mice by their cytokine production and were also able to separate mice with moderate-to-high cytokine production by infection strain. The data presented here show that Coccidioides and/or the host produce volatile metabolites that may yield biomarkers for a Valley fever breath test that can detect coccidioidal infection and provide clinically relevant information on primary pulmonary disease severity.

Abstract Background The above-average proportion of people with opioid use disorder living in prisons is a worldwide reality, and the need to treat these people was recognized internationally more than 20 years ago. Studies have shown that substitution therapies are best suited to treat opioid use disorder and reduce the risk of HIV and hepatitis C transmission and overdose. However, huge health inequalities exist in and outside of prison due to the different implementation of opioid substitution therapy (OST). People living in prisons are entitled to the best possible health care. This is established by the Universal Declaration of Human Rights and by the International Convention on Economic, Social and Cultural Rights. Solely the imprisonment, and not the loss of fundamental human rights, constitutes the punishment. Methods A qualitative literature search using PubMed and Google Scholar was performed in order to identify relevant publications. Results This review shows the inequality in availability of opioid substitution therapy for people living in prison compared with people outside of prison in Germany. It also gives possible reasons and evidence for this inequality, showing that continuing or initiating OST in prison is more beneficial for the health of people living in prison than abstinence-oriented treatment only. Conclusion It is important that drug use disorder is treated as a serious illness also in prison. Joint efforts are needed to provide people living in prison with the best possible treatment and to minimize the adverse effects of drug use. Therefore, with laws, policies, and programs that conform to international human rights standards, each state must ensure that people living in prison receive the same health care as people outside of prison.

DNA composite materials have potential for biomedical sciences; however, control over the materials can be an issue. Here, the authors report on a carbon-nanotube reinforced DNA-silica gel with controllable mechanical properties to steer the attachment, proliferation, migration and release of cells.

Mammalian cells release different types of vesicles, collectively termed extracellular vesicles (EVs). EVs contain cellular microRNAs (miRNAs) with an apparent potential to deliver their miRNA cargo to recipient cells to affect the stability of individual mRNAs and the cells' transcriptome. The extent to which miRNAs are exported via the EV route and whether they contribute to cell-cell communication are controversial. To address these issues, we defined multiple properties of EVs and analyzed their capacity to deliver packaged miRNAs into target cells to exert biological functions. We applied well-defined approaches to produce and characterize purified EVs with or without specific viral miRNAs. We found that only a small fraction of EVs carried miRNAs. EVs readily bound to different target cell types, but EVs did not fuse detectably with cellular membranes to deliver their cargo. We engineered EVs to be fusogenic and document their capacity to deliver functional messenger RNAs. Engineered fusogenic EVs, however, did not detectably alter the functionality of cells exposed to miRNA-carrying EVs. These results suggest that EV-borne miRNAs do not act as effectors of cell-to-cell communication.