Comparison of selectivity and sensitivity of various ferric selective electrodes prepared using N-((bis(dimethyl amino)methylene)carbamothioyl)benzamide.

N-((Bis(dimethyl amino)methylene)carbamothioyl)benzamide (NBMCB) was synthesized, characterized, and used as an ionophore for producing three novel ion-selective potentiometric sensors for Fe(III) determination. Firstly, using the molecular mechanic-based MMFF94 method, the most stable NBMCB's conformer and its isosteric complexes with various cations were determined. According to the Gibbs free energy results of the reaction, the thermodynamic complexation reactivity of Fe(III) and the ligand was acceptable. These results were obtained using the B3LYP approach and the 6-31G(d,p) basis set that was substituted for heavy metals by the LanL2DZ basis set. We used UV-visible spectrophotometry to confirm the tendency of NBMCB to react with Fe(III). Generally, three diverse liquid membrane ferric selective electrodes were obtained by the use of the specified ligand: classic with a liquid internal electrolyte-ferric selective electrode (LIE-FSE), solid state-FSE (SS-FSE), and coated wire-FSE (CW-FSE). The reactions exhibited Nernstian behavior across all electrodes. The limit of detection was enhanced for the SS-FSE (3 × 10-9 M) and the CW-FSE (3 × 10-7 M) in comparison with that of the LIE-FSE (7 × 10-7 M). The lifetime of the LIE-FSE was 8 weeks, while it was 10 weeks for the SS-FSE and the CW-FSE. Elimination of the internal solution reduced the limit of detection and prolonged the lifespan of the sensors. Also, the three electrodes all had a short response time of around 5-7 s. The sensors were utilized as indicator electrodes during the potentiometric titration of Fe(III) using ethylenediaminetetraacetic acid.

Activated Gab1 drives hepatocyte proliferation and anti-apoptosis in liver fibrosis via potential involvement of the HGF/c-Met signaling axis.

Chronic liver diseases are caused by hepatic viral infection, chemicals, and metabolic stress. The protein Grb2-associated binder 1 (Gab1) binds to various growth factor receptors, and triggers cell differentiation/survival signaling pathways. To identify signaling molecules involved in the progression of liver diseases, we performed reverse-phase protein microarray (RPMA)-based screening of hepatocytes isolated from humanized mice after acute HCV infection. Acute viral infection in humanized liver mice significantly decreased the level of hepatocyte p-Gab1. Moreover, hepatoma cells upon HCV infection decreased Gab1 mRNA at later times of infection (D3 to D5) and p-Gab1 level was inversely related to the production of TGF-ß. In contrast, the level of p-Gab1 was increased in CCL4-induced fibrotic liver. Hepatoma cells showed elevation of p-Gab1, along with an increase in STAT3 and ERK activation, upon treatment with HGF (ligand of HGF receptor/c-Met) and CCL4. In Gab1 knockdown hepatoma cells, cell proliferative signaling activity was reduced but the level of activated caspase-3 was increased. These findings suggest that hepatocyte Gab1 expression may play a role in promoting liver fibrosis progression by triggering ERK activation and inhibiting apoptosis. It implies that the Gab1-mediated signaling pathway would be a promising therapeutic target to treat chronic liver diseases.

HSP90 is part of a protein complex with the L polymerase of Rift Valley fever phlebovirus and prevents its degradation by the proteasome during the viral genome replication/transcription stage.

The mosquito-borne Rift Valley fever virus (RVFV) from the Phenuiviridae family is a single-stranded RNA virus that causes the re-emerging zoonotic disease Rift Valley fever (RVF). Classified as a Category A agent by the NIH, RVFV infection can cause debilitating disease or death in humans and lead to devastating economic impacts by causing abortion storms in pregnant cattle. In a previous study, we showed that the host chaperone protein HSP90 is an RVFV-associated host factor that plays a critical role post viral entry, during the active phase of viral genome replication/transcription. In this study, we have elucidated the molecular mechanisms behind the regulatory effect of HSP90 during infection with RVFV. Our results demonstrate that during the early infection phase, host HSP90 associates with the viral RNA-dependent RNA polymerase (L protein) and prevents its degradation through the proteasome, resulting in increased viral replication.

A meta-analysis of methane-mitigation potential of feed additives evaluated in vitro.

A systematic literature review of in vitro studies was performed to identify methane (CH4) mitigation interventions with a potential to reduce CH4 emission in vivo. Data from 277 peer-reviewed studies published between 1979 and 2018 were reviewed. Individual CH4 mitigation interventions were classified into 14 categories of feed additives based on their type, chemical composition, and mode of action. Response variables evaluated were absolute CH4 emission (number of treatment means comparisons = 1,325); total volatile fatty acids (n = 1,007), acetate (n = 783), propionate (n = 792), and butyrate (n = 776) concentrations; acetate to propionate ratio (n = 675); digestibility of dry matter (n = 489), organic matter (n = 277), and neutral detergent fiber (n = 177). Total gas production was used as an explanatory variable in the model for CH4 production. Relative mean difference between treatment and control means reported in the studies was calculated and used for statistical analysis. The robust variance estimation method was used to analyze the effects of CH4 mitigation interventions. In vitro CH4 production was decreased by antibodies (-38.9%), chemical inhibitors (-29.2%), electron sinks (-18.9%), essential oils (-18.2%), plant extracts (-14.5%), plant inclusion (-11.7%), saponins (-14.8%), and tannins (-14.5%). Overall effects of direct-fed microbials, enzymes, macroalgae, and organic acids supplementation did not affect CH4 production in the current meta-analysis. When considering the effects of individual mitigation interventions containing a minimum number of 4 degrees of freedom within feed additives categories, Enterococcus spp. (i.e., direct-fed microbial), nitrophenol (i.e., electron sink), and Leucaena spp. (i.e., tannins) decreased CH4 production by 20.3%, 27.1%, and 23.5%, respectively, without extensively, or only slightly, affecting ruminal fermentation and digestibility of nutrients. It should be noted, however, that although the total number of publications (n = 277) and treatment means comparisons (n = 1,325 for CH4 production) in the current analysis were high, data for most mitigation interventions were obtained from less than 5 observations (e.g., maximum number of observations was 4, 7, and 22 for nitrophenol, Enterococcus spp., and Leucaena spp., respectively), because of limited data available in the literature. These should be further evaluated in vitro and in vivo to determine their true potential to decrease enteric CH4 production, yield, and intensity. Some mitigation interventions (e.g., magnesium, Heracleum spp., nitroglycerin, ß-cyclodextrin, Leptospermum pattersoni, Fructulus Ligustri, Salix caprea, and Sesbania grandiflora) decreased in vitro CH4 production by over 50% but did not have enough observations in the database. These should be more extensively investigated in vitro, and the dose effect must be considered before adoption of mitigation interventions in vivo.

Effects of herd management decisions on dairy cow longevity, farm profitability, and emissions of enteric methane - a simulation study of milk and beef production.

Sustainable dairy and beef production provides environmental, economic, and social values that can potentially be maximized by optimizing herd management strategies. The length of a dairy cow's life is affected by, and affects, all three pillars of sustainability. Longevity in dairy cows is multifactorial and strongly dependent on herd management. Despite genetic improvements, the average time of culling for Swedish cows has barely changed and is currently at 2.6 lactations. This culling rate requires a high number of replacement heifers, generating high rearing costs for farmers. This study evaluated different herd management strategies to improve cow longevity and assessed the effects on enteric methane (CH4) emissions from the herd and the profitability of milk production and beef production from the dairy cows and their offspring. The base scenario, an average Swedish Holstein herd of 100 cows, was compared with seven scenarios simulated using a stochastic herd simulation model (SimHerd). Two of these scenarios involved improved health and survival of cows in the herd, three involved improved reproduction, one considered the consequences of keeping all surplus heifers in the herd, and one considered maximizing the use of X-sorted dairy semen and inseminating the rest of the herd with unsorted beef semen, to avoid surplus replacement heifers. Improved fertility had the greatest effect in increasing the productive life per cow, to 3.8 years compared with 2.8 in the base scenario, allowed for more use of beef semen, reduced the number of replacement heifers, and generated the highest herd profit (€98 per cow-year higher than base scenario). Keeping all surplus heifers instead of producing beef × dairy cross calves decreased the number of productive years by 0.8 and reduced profit by €22 per cow-year. The profit was highly associated with costs related to replacement heifers. The highest beef output (3 369 kg per year more than base scenario) was achieved by keeping all heifers and culling a high share of dairy cows, but this scenario also generated much higher enteric CH4 emissions (+1 257 kg per year). Improving health, survival, or fertility reduced enteric CH4 emissions by 90-255 kg per year, while total yearly beef production ranged from 59 kg less to 556 kg more than in the base scenario. Reducing the number of replacement heifers needed by improving cow reproductive performance is thus key to increasing cow longevity and profitability, while reducing enteric CH4 emissions from the herd without compromising milk and meat production.

Redox-Mediated Rewiring of Signalling Pathways: The Role of a Cellular Clock in Brain Health and Disease.

Metazoan signalling pathways can be rewired to dampen or amplify the rate of events, such as those that occur in development and aging. Given that a linear network topology restricts the capacity to rewire signalling pathways, such scalability of the pace of biological events suggests the existence of programmable non-linear elements in the underlying signalling pathways. Here, we review the network topology of key signalling pathways with a focus on redox-sensitive proteins, including PTEN and Ras GTPase, that reshape the connectivity profile of signalling pathways in response to an altered redox state. While this network-level impact of redox is achieved by the modulation of individual redox-sensitive proteins, it is the population by these proteins of critical nodes in a network topology of signal transduction pathways that amplifies the impact of redox-mediated reprogramming. We propose that redox-mediated rewiring is essential to regulate the rate of transmission of biological signals, giving rise to a programmable cellular clock that orchestrates the pace of biological phenomena such as development and aging. We further review the evidence that an aberrant redox-mediated modulation of output of the cellular clock contributes to the emergence of pathological conditions affecting the human brain.

A meta-analysis of methane mitigation potential of feed additives evaluated in vitro.

A systematic literature review of in vitro studies was performed to identify methane (CH4) mitigation interventions with a potential to reduce CH4 emission in vivo. Data from 277 peer-reviewed studies published between 1979 and 2018 were reviewed. Individual CH4 mitigation interventions were classified into 14 categories of feed additives based on their type, chemical composition, and mode of action. Response variables evaluated were absolute CH4 emission (number of treatment means comparisons = 1,325); total volatile fatty acids (VFA; n = 1,007), acetate (n = 783), propionate (n = 792), and butyrate (n = 776) concentrations; acetate to propionate ratio (A:P; n = 675); digestibility of dry matter (DM; n = 489), organic matter (OM; n = 277), and neutral detergent fiber (NDF; n = 177). Total gas production was used as an explanatory variable in the model for CH4 production. Relative mean difference between treatment and control means reported in the studies were calculated and used for statistical analysis. Robust variance estimation method was used to analyze the effects of CH4 mitigation interventions. In vitro CH4 production was decreased by antibodies (-38.9%), chemical inhibitors (-29.2%), electron sinks (-18.9%), essential oils (-18.2%), plant extracts (-14.5%), plants inclusion (-11.7%), saponins (-14.8%), and tannins (-14.5%). Overall effects of direct fed microbials, enzymes, macroalgae, and organic acids supplementation did not affect CH4 production in the current meta-analysis. When considering the effects of individual mitigation interventions containing a minimum number of 4 degrees of freedom within feed additives categories, Enterococcus spp. (i.e., direct fed microbial), nitrophenol (i.e., electron sink), and Leucaena spp. (i.e., tannins) decreased CH4 production by 20.3, 27.1, and 23.5%, respectively, without extensively, or only slightly, affecting ruminal fermentation and digestibility of nutrients. It should be noted, however, that although the total number of publications (n = 277) and treatment means comparisons (n = 1,325 for CH4 production) in the current analysis were high, data for most mitigation interventions were obtained from less than 5 observations (e.g., maximum number of observations was 4, 7, and 22 for nitrophenol, Enterococcus spp., and Leucaena spp., respectively), because of limited data available in the literature. These should be further evaluated in vitro and in vivo to determine their true potential to decrease enteric CH4 production, yield, and intensity. Some mitigation interventions (e.g., magnesium, Heracleum spp., nitroglycerin, ß-cyclodextrin, Leptospermum pattersoni, Fructulus Ligustri, Salix caprea, and Sesbania grandiflora) decreased in vitro CH4 production by over 50% but did not have enough observations in the database. These should be more extensively investigated in vitro, and the dose effect must be considered before adoption of mitigation interventions in vivo.

Effects of replacement of barley with oats on milk fatty acid composition in dairy cows fed grass silage-based diets.

This study consists of milk fatty acid (FA) data collected during 2 in vivo experiments. For this study, 8 cows from each experiment were included in a replicated 4 × 4 Latin square design. At the start of experiment 1 (Exp1) cows were at (mean ± standard deviation) 87 ± 34.6 d in milk, 625 ± 85.0 kg of body weight, and 32.1 ± 4.17 kg/d milk yield and at the start of experiment 2 (Exp2) cows were at 74 ± 18.2 d in milk, 629 ± 87.0 kg of body weight, and 37.0 ± 3.2 kg/d milk yield. In Exp1, we examined the effects of gradual replacement of barley with hulled oats (oats with hulls) on milk FA composition. The basal diet was grass silage and rapeseed meal (58 and 10% of diet DM, respectively), and the 4 grain supplements were formulated so that barley was gradually replaced by hulled oats at levels of 0, 33, 67, and 100% on dry matter basis. In Exp2, we examined (1) the effects of replacing barley with both hulled and dehulled oats (oats without hulls) and (2) the effects of gradual replacement of hulled oats with dehulled oats on milk FA composition. The basal diet was grass silage and rapeseed meal (60 and 10% of diet DM, respectively), and the 4 pelleted experimental concentrates were barley, hulled oats, a 50:50 mixture of hulled and dehulled oats, and dehulled oats on dry matter basis. In Exp1, gradual replacement of barley with hulled oats decreased relative proportions of 14:0, 16:0, and total saturated FA (SFA) in milk fat linearly, whereas proportions of 18:0, 18:1, total monounsaturated FA, and total cis unsaturated FA increased linearly. Transfer efficiency of total C18 decreased linearly when barley was replaced by hulled oats in Exp1. In Exp2, relative proportions of 14:0, 16:0, and total SFA were lower, whereas proportions of 18:0, 18:1, monounsaturated FA, and cis unsaturated FA were higher in milk from cows fed the oat diets than in milk from cows fed the barley diet. Moreover, in Exp2, gradual replacement of hulled oats with dehulled oats slightly decreased the relative proportion of 14:0 in milk fat but did not affect the proportions of 16:0, 18:0, 18:1, total SFA, monounsaturated FA, trans FA, or polyunsaturated FA. In Exp2, transfer efficiency of total C18 was lower when cows were fed the oat diets than when fed the barley diet and decreased linearly when hulled oats were replaced with dehulled oats. Predictions of daily CH4 emissions (g/d) using the on-farm available variables energy-corrected milk yield and body weight were not markedly improved by including milk concentrations of individual milk FA in prediction equations. In conclusion, replacement of barley with oats as a concentrate supplement for dairy cows fed a grass silage-based diet could offer a practical strategy to change the FA composition of milk to be more in accordance with international dietary guidelines regarding consumption of SFA.

Cellular self-organization: An overdrive in Cambrian diversity?

During the early Cambrian period metazoan life forms diverged at an accelerated rate to occupy multiple ecological niches on earth. A variety of explanations have been proposed to address this major evolutionary phenomenon termed the "Cambrian explosion." While most hypotheses address environmental, developmental, and ecological factors that facilitated evolutionary innovations, the biological basis for accelerated emergence of species diversity in the Cambrian period remains largely conjectural. Herein, we posit that morphogenesis by self-organization enables the uncoupling of genomic mutational landscape from phenotypic diversification. Evidence is provided for a two-tiered interpretation of genomic changes in metazoan animals wherein mutations not only impact upon function of individual cells, but also alter the self-organization outcome during developmental morphogenesis. We provide evidence that the morphological impacts of mutations on self-organization could remain repressed if associated with an unmet negative energetic cost. We posit that accelerated morphological diversification in transition to the Cambrian period has occurred by emergence of dormant (i.e., reserved) morphological novelties whose molecular underpinnings were seeded in the Precambrian period.

Notch ankyrin domain: evolutionary rise of a thermodynamic sensor.

Notch signalling pathway plays a key role in metazoan biology by contributing to resolution of binary decisions in the life cycle of cells during development. Outcomes such as proliferation/differentiation dichotomy are resolved by transcriptional remodelling that follows a switch from Notchon to Notchoff state, characterised by dissociation of Notch intracellular domain (NICD) from DNA-bound RBPJ. Here we provide evidence that transitioning to the Notchoff state is regulated by heat flux, a phenomenon that aligns resolution of fate dichotomies to mitochondrial activity. A combination of phylogenetic analysis and computational biochemistry was utilised to disclose structural adaptations of Notch1 ankyrin domain that enabled function as a sensor of heat flux. We then employed DNA-based micro-thermography to measure heat flux during brain development, followed by analysis in vitro of the temperature-dependent behaviour of Notch1 in mouse neural progenitor cells. The structural capacity of NICD to operate as a thermodynamic sensor in metazoans stems from characteristic enrichment of charged acidic amino acids in ß-hairpins of the ankyrin domain that amplify destabilising inter-residue electrostatic interactions and render the domain thermolabile. The instability emerges upon mitochondrial activity which raises the perinuclear and nuclear temperatures to 50 °C and 39 °C, respectively, leading to destabilization of Notch1 transcriptional complex and transitioning to the Notchoff state. Notch1 functions a metazoan thermodynamic sensor that is switched on by intercellular contacts, inputs heat flux as a proxy for mitochondrial activity in the Notchon state via the ankyrin domain and is eventually switched off in a temperature-dependent manner. Video abstract.

Mitochondrial Extracellular Vesicles in CNS Disorders: New Frontiers in Understanding the Neurological Disorders of the Brain.

Recent findings have highlighted potential diagnostic and prognostic values of extracellular vesicles (EVs) that contain mitochondrial derived components for neurological disorders. Furthermore, functional influences of vesicles carrying mitochondrial components have been reported. In particular, this includes indications of crosstalk with mitophagy to influence progression of various CNS disorders. In this mini-review, we discuss the current state of knowledge about this intriguing class of vesicles in neurological disorders of the CNS, and outline the lacunae and thus scope of further development in this fascinating field of study.

A Microfluidic Platform to Monitor Real-Time Effects of Extracellular Vesicle Exchange between Co-Cultured Cells across Selectively Permeable Barriers.

Exosomes and other extracellular vesicles (EVs) play a significant yet poorly understood role in cell-cell communication during homeostasis and various pathological conditions. Conventional in vitro and in vivo approaches for studying exosome/EV function depend on time-consuming and expensive vesicle purification methods to obtain sufficient vesicle populations. Moreover, the existence of various EV subtypes with distinct functional characteristics and submicron size makes their analysis challenging. To help address these challenges, we present here a unique chip-based approach for real-time monitoring of cellular EV exchange between physically separated cell populations. The extracellular matrix (ECM)-mimicking Matrigel is used to physically separate cell populations confined within microchannels, and mimics tissue environments to enable direct study of exosome/EV function. The submicron effective pore size of the Matrigel allows for the selective diffusion of only exosomes and other smaller EVs, in addition to soluble factors, between co-cultured cell populations. Furthermore, the use of PEGDA hydrogel with a very small pore size of 1.2 nm in lieu of Matrigel allows us to block EV migration and, therefore, differentiate EV effects from effects that may be mediated by soluble factors. This versatile platform bridges purely in vitro and in vivo assays by enabling studies of EV-mediated cellular crosstalk under physiologically relevant conditions, enabling future exosome/EV investigations across multiple disciplines through real-time monitoring of vesicle exchange.

Neural pericytes: A genomic archival state programmed by CHromatin topology.

Accumulating evidence suggests that mural pericytes, apart from stabilizing the associated microvessels, play additional roles in regeneration of local cellular elements. Herein, the mechanistic basis for such diverse and at times contradictory roles adopted by pericytes in the brain is reviewed. Core concepts of an emerging model are discussed wherein mural pericytes reside in a metastable "archival state" that conceals a neural progenitor identity. Upon angiogenic remodeling, a selected subpopulation of pericytes reclaim the progenitor state during transdifferentiation and contribute to neural regeneration. The genomic basis for neural transdifferentiation of pericytes is reviewed with reference to the extant literature.

Comparison of Molly and Karoline models to predict methane production in growing and dairy cattle.

Numerous empirical and mechanistic models predicting methane (CH4) production are available. The aim of this work was to evaluate the Molly cow model and the Nordic cow model Karoline in predicting CH4 production in cattle using a data set consisting of 267 treatment means from 55 respiration chamber studies. The dietary and animal characteristics used for the model evaluation represent the range of diets fed to dairy and growing cattle. Feedlot diets and diets containing additives mitigating CH4 production were not included in the data set. The relationships between observed and predicted CH4 (pCH4) were assessed by regression analysis using fixed and mixed model analysis. Residual analysis was conducted to evaluate which dietary factors were related to prediction errors. The fixed model analysis showed that the Molly predictions were related to the observed data (± standard error) as CH4 (g/d) = 0.94 (±0.022) × pCH4 (g/d) + 31 (±6.9) [root mean squared prediction error (RMSPE) = 45.0 g/d (14.9% of observed mean), concordance correlation coefficient (CCC) = 0.925]. The corresponding equation for the Karoline model was CH4 (g/d) = CH4 (g/d) = 0.98 (±0.019) × pCH4 (g/d) + 7.0 (±6.0) [RMSPE = 35.0 g/d (11.6%), CCC = 0.953]. Proportions of mean squared prediction error attributable to mean and linear bias and random error were 10.6, 2.2, and 87.2% for the Molly model, and 1.3, 0.3, and 98.6% for the Karoline model, respectively. Mean and linear bias were significant for the Molly model but not for the Karoline model. With the mixed model regression analysis RMSPE adjusted for random study effects were 10.9 and 7.9% for the Molly model and the Karoline model, respectively. The residuals of CH4 predictions were more strongly related to factors associated with CH4 production (feeding level, digestibility, fat concentrations) with the Molly model compared with the Karoline model. Especially large mean (underprediction) and linear bias (overprediction of low digestibility diets relative to high digestibility diets) contributed to the prediction error of CH4 yield with the Molly model. It was concluded that both models could be used for prediction of CH4 production in cattle, but Karoline was more accurate and precise based on smaller RMSPE, mean bias, and slope bias, and greater CCC. The importance of accurate input data of key variables affecting diet digestibility is emphasized.

Programmed genomic instability regulates neural transdifferentiation of human brain microvascular pericytes.

BACKGROUND: Transdifferentiation describes transformation in vivo of specialized cells from one lineage into another. While there is extensive literature on forced induction of lineage reprogramming in vitro, endogenous mechanisms that govern transdifferentiation remain largely unknown. The observation that human microvascular pericytes transdifferentiate into neurons provided an opportunity to explore the endogenous molecular basis for lineage reprogramming. RESULTS: We show that abrupt destabilization of the higher-order chromatin topology that chaperones lineage memory of pericytes is driven by transient global transcriptional arrest. This leads within minutes to localized decompression of the repressed competing higher-order chromatin topology and expression of pro-neural genes. Transition to neural lineage is completed by probabilistic induction of R-loops in key myogenic loci upon re-initiation of RNA polymerase activity, leading to depletion of the myogenic transcriptome and emergence of the neurogenic transcriptome. CONCLUSIONS: These findings suggest that the global transcriptional landscape not only shapes the functional cellular identity of pericytes, but also stabilizes lineage memory by silencing the competing neural program within a repressed chromatin state.

Exosomes originating from infection with the cytoplasmic single-stranded RNA virus Rift Valley fever virus (RVFV) protect recipient cells by inducing RIG-I mediated IFN-B response that leads to activation of autophagy.

BACKGROUND: Although multiple studies have demonstrated a role for exosomes during virus infections, our understanding of the mechanisms by which exosome exchange regulates immune response during viral infections and affects viral pathogenesis is still in its infancy. In particular, very little is known for cytoplasmic single-stranded RNA viruses such as SARS-CoV-2 and Rift Valley fever virus (RVFV). We have used RVFV infection as a model for cytoplasmic single-stranded RNA viruses to address this gap in knowledge. RVFV is a highly pathogenic agent that causes RVF, a zoonotic disease for which no effective therapeutic or approved human vaccine exist. RESULTS: We show here that exosomes released from cells infected with RVFV (designated as EXi-RVFV) serve a protective role for the host and provide a mechanistic model for these effects. Our results show that treatment of both naïve immune cells (U937 monocytes) and naïve non-immune cells (HSAECs) with EXi-RVFV induces a strong RIG-I dependent activation of IFN-B. We also demonstrate that this strong anti-viral response leads to activation of autophagy in treated cells and correlates with resistance to subsequent viral infection. Since we have shown that viral RNA genome is associated with EXi-RVFV, RIG-I activation might be mediated by the presence of packaged viral RNA sequences. CONCLUSIONS: Using RVFV infection as a model for cytoplasmic single-stranded RNA viruses, our results show a novel mechanism of host protection by exosomes released from infected cells (EXi) whereby the EXi activate RIG-I to induce IFN-dependent activation of autophagy in naïve recipient cells including monocytes. Because monocytes serve as reservoirs for RVFV replication, this EXi-RVFV-induced activation of autophagy in monocytes may work to slow down or halt viral dissemination in the infected organism. These findings offer novel mechanistic insights that may aid in future development of effective vaccines or therapeutics, and that may be applicable for a better molecular understanding of how exosome release regulates innate immune response to other cytoplasmic single-stranded RNA viruses.

Replacement of barley with oats and dehulled oats: Effects on milk production, enteric methane emissions, and energy utilization in dairy cows fed a grass silage-based diet.

Sixteen Nordic Red dairy cows, at 80 ± 4.6 d in milk and with an average body weight of 624 ± 91.8 kg, were used in a replicated 4 × 4 Latin square design to investigate the effects of different concentrate supplements on milk production, enteric CH4 emissions, ruminal fermentation, digestibility, and energy utilization. The cows were blocked into 4 groups based on parity and milk yield and randomly assigned to 1 of 4 experimental concentrates: (1) barley, (2) hulled oats, (3) an oat mixture consisting of hulled and dehulled oats, 50:50 on dry matter basis, and (4) dehulled oats; canola meal was a protein supplement in all 4 concentrates. The cows were fed grass silage and experimental concentrate (forage-to-concentrate ratio 60:40 on dry matter basis) ad libitum. To compare the effects of barley and oats, the barley diet was compared with the overall mean of the hulled oat, oat mixture, and dehulled oat diets. To investigate the effects of gradual replacement of hulled oats with dehulled oats, linear and quadratic contrasts were specified. Milk and energy-corrected milk (ECM) yield were higher on the oat diets compared with the barley diet but were not affected by the type of oats. Concentrations of milk constituents were not affected by grain species or type of oats, except for protein concentration, which was lower on the oat diets than on the barley diet. Feeding the oat diets led to higher milk protein yield and higher milk urea N concentrations. Feed efficiency tended to be higher on the oat diets, and linearly increased with increased inclusion of dehulled oats. Methane emissions (g/d) and CH4 yield (g/kg of dry matter intake) were unaffected by grain species but increased linearly with increasing inclusion of dehulled oats in the diet. Because of higher ECM yield, CH4 intensity (g/kg of ECM) was on average 5.7% lower from cows on the oat diets than on the barley diet. Ruminal fermentation was not affected by dietary treatment. Total-tract apparent digestibility of organic matter, crude protein, and neutral detergent fiber was unaffected by grain species but linearly increased with increasing inclusion of dehulled oats. Gross energy content was higher on the oat diets and linearly increased with increasing inclusion of dehulled oats. Feeding the oat diets led to a lower ratio of CH4 energy to gross energy intake, greater milk energy and heat production but no change in energy balance. Gradual replacement of hulled oats with dehulled oats linearly increased gross energy digestibility, CH4 energy, metabolizable energy intake, heat production, and energy balance. We observed no effect of dietary treatment on efficiency of metabolizable energy use for lactation. In conclusion, replacing barley with any type of oats increased milk and ECM yield, which led to a 5.7% decrease in CH4 intensity. In addition, dehulling of oats before feeding is unnecessary because it did not significantly improve production performance of dairy cows in positive energy balance.

The fate of notch-1 transcript is linked to cell cycle dynamics by activity of a natural antisense transcript.

A core imprint of metazoan life is that perturbations of cell cycle are offset by compensatory changes in successive cellular generations. This trait enhances robustness of multicellular growth and requires transmission of signaling cues within a cell lineage. Notably, the identity and mode of activity of transgenerational signals remain largely unknown. Here we report the discovery of a natural antisense transcript encoded in exon 25 of notch-1 locus (nAS25) by which mother cells control the fate of notch-1 transcript in daughter cells to buffer against perturbations of cell cycle. The antisense transcript is transcribed at G1 phase of cell cycle from a bi-directional E2F1-dependent promoter in the mother cell where the titer of nAS25 is calibrated to the length of G1. Transmission of the antisense transcript from mother to daughter cells stabilizes notch-1 sense transcript in G0 phase of daughter cells by masking it from RNA editing and resultant nonsense-mediated degradation. In consequence, nAS25-mediated amplification of notch-1 signaling reprograms G1 phase in daughter cells to compensate for the altered dynamics of the mother cell. The function of nAS25/notch-1 in integrating G1 phase history of the mother cell into that of daughter cells is compatible with the predicted activity of a molecular oscillator, slower than cyclins, that coordinates cell cycle within cell lineage.

A traditional medicine, respiratory detox shot (RDS), inhibits the infection of SARS-CoV, SARS-CoV-2, and the influenza A virus in vitro.

BACKGROUND: The ongoing global pandemic of coronavirus disease 2019 (COVID-19) has resulted in the infection of over 128 million people and has caused over 2.8 million deaths as of April 2021 in more than 220 countries and territories. Currently, there is no effective treatment for COVID-19 to reduce mortality. We investigated the potential anti-coronavirus activities from an oral liquid of traditional medicine, Respiratory Detox Shot (RDS), which contains mostly herbal ingredients traditionally used to manage lung diseases. RESULTS: Here we report that RDS inhibited the infection of target cells by lenti-SARS-CoV, lenti-SARS-CoV-2, and hybrid alphavirus-SARS-CoV-2 (Ha-CoV-2) pseudoviruses, and by infectious SARS-CoV-2 and derived Ha-CoV-2 variants including B.1.1.7, B.1.351, P.1, B.1.429, B.1.2, B.1.494, B.1.1.207, B.1.258, and B.1.1.298. We further demonstrated that RDS directly inactivates the infectivity of SARS-CoV-2 virus particles. In addition, we found that RDS can also block the infection of target cells by Influenza A virus. CONCLUSIONS: These results suggest that RDS may broadly inhibit the infection of respiratory viruses.

Cannibalized erythroblasts accelerate developmental neurogenesis by regulating mitochondrial dynamics.

Metabolic support was long considered to be the only developmental function of hematopoiesis, a view that is gradually changing. Here, we disclose a mechanism triggered during neurulation that programs brain development by donation of sacrificial yolk sac erythroblasts to neuroepithelial cells. At embryonic day (E) 8.5, neuroepithelial cells transiently integrate with the endothelium of yolk sac blood vessels and cannibalize intravascular erythroblasts as transient heme-rich endosymbionts. This cannibalistic behavior instructs precocious neuronal differentiation of neuroepithelial cells in the proximity of blood vessels. By experiments in vitro, we show that access to erythroblastic heme accelerates the pace of neurogenesis by induction of a truncated neurogenic differentiation program from a poised state. Mechanistically, the poised state is invoked by activation of the mitochondrial electron transport chain that leads to amplified production of reactive oxygen species in addition to omnipresent guanosine triphosphate (GTP) with consequential upregulation of pro-differentiation ß-catenin.