The putative RNA helicase DDX1 associates with the nuclear RNA exosome and modulates RNA/DNA hybrids (R-loops).

The RNA exosome is a ribonuclease complex that mediates both RNA processing and degradation. This complex is evolutionarily conserved, ubiquitously expressed, and required for fundamental cellular functions, including rRNA processing. The RNA exosome plays roles in regulating gene expression and protecting the genome, including modulating the accumulation of RNA-DNA hybrids (R-loops). The function of the RNA exosome is facilitated by cofactors, such as the RNA helicase MTR4, which binds/remodels RNAs. Recently, missense mutations in RNA exosome subunit genes have been linked to neurological diseases. One possibility to explain why missense mutations in genes encoding RNA exosome subunits lead to neurological diseases is that the complex may interact with cell- or tissue-specific cofactors that are impacted by these changes. To begin addressing this question, we performed immunoprecipitation of the RNA exosome subunit, EXOSC3, in a neuronal cell line (N2A), followed by proteomic analyses to identify novel interactors. We identified the putative RNA helicase, DDX1, as an interactor. DDX1 plays roles in double-strand break repair, rRNA processing, and R-loop modulation. To explore the functional connections between EXOSC3 and DDX1, we examined the interaction following double-strand breaks and analyzed changes in R-loops in N2A cells depleted for EXOSC3 or DDX1 by DNA/RNA immunoprecipitation followed by sequencing. We find that EXOSC3 interaction with DDX1 is decreased in the presence of DNA damage and that loss of EXOSC3 or DDX1 alters R-loops. These results suggest EXOSC3 and DDX1 interact during events of cellular homeostasis and potentially suppress unscrupulous expression of genes promoting neuronal projection.

Best practices for the visualization, mapping, and manipulation of R-loops.

R-loops represent an abundant class of large non-B DNA structures in genomes. Even though they form transiently and at modest frequencies, interfering with R-loop formation or dissolution has significant impacts on genome stability. Addressing the mechanism(s) of R-loop-mediated genome destabilization requires a precise characterization of their distribution in genomes. A number of independent methods have been developed to visualize and map R-loops, but their results are at times discordant, leading to confusion. Here, we review the main existing methodologies for R-loop mapping and assess their limitations as well as the robustness of existing datasets. We offer a set of best practices to improve the reproducibility of maps, hoping that such guidelines could be useful for authors and referees alike. Finally, we propose a possible resolution for the apparent contradictions in R-loop mapping outcomes between antibody-based and RNase H1-based mapping approaches.

Optimizing wheat productivity through integrated management of irrigation, nutrition, and organic amendments.

Enhancing wheat productivity by implementing a comprehensive approach that combines irrigation, nutrition, and organic amendments shows potential for collectively enhancing crop performance. This study examined the individual and combined effects of using irrigation systems (IS), foliar potassium bicarbonate (PBR) application, and compost application methods (CM) on nine traits related to the growth, physiology, and yield of the Giza-171 wheat cultivar. Analysis of variance revealed significant (P ≤ 0.05) main effects of IS, PBR, and CM on wheat growth, physiology, and yield traits over the two growing seasons of the study. Drip irrigation resulted in a 16% increase in plant height, leaf area index, crop growth rate, yield components, and grain yield compared to spray irrigation. Additionally, the application of foliar PBR at a concentration of 0.08 g/L boosted these parameters by up to 22% compared to the control. Furthermore, the application of compost using the role method resulted in enhanced wheat performance compared to the treatment including mix application. Importantly, the combined analysis revealed that the three-way interaction between the three factors had a significant effect (P ≤ 0.05) on all the studied traits, with drip irrigation at 0.08 g PBR rate and role compost application method (referred as Drip_0.08g_Role) resulting in the best performance across all traits, while sprinkle irrigation without PBR and conventional mixed compost method (referred as sprinkle_CK_Mix) produced the poorest results. This highlights the potential to synergistically improve wheat performance through optimized agronomic inputs.

Efficiency dynamics among onion growers in Maharashtra: a comparative analysis of drip irrigation adopters and non-adopters.

BACKGROUND: Onions are economically and nutritionally important vegetable crops. Despite advances in technology and acreage, Indian onion growers face challenges in realizing their full productivity potential. This study examines the technical efficiency of onion growers, the factors influencing it, and the constraints faced by those adopting drip irrigation in the Ghod river basin of western Maharashtra. A sample of 480 farmers including those practicing drip irrigation and those not practicing it, was selected from Junnar, Shirur, Parner, and Shrigonda blocks of the basin. The primary data was collected through semi-structured interviews. Analytical tools such as the Cobb-Douglas production function (represents technological relationship between multiple inputs and the resulting output), a single-stage stochastic frontier model, the Tobit model, and descriptive statistics were used to assess the technical efficiency of onion production at the farm level. RESULTS: According to the maximum likelihood estimates of the stochastic frontier analysis, drip adopters exhibited a mean technical efficiency of 92%, while for non-adopters it was 65%. It indicates that the use of drip irrigation technology is associated with higher technical efficiency. The association of technical efficiency and socio-economic characters of households showed that education, extension contacts, social participation, and use of information sources had a positive influence on technical efficiency, while family size had a negative influence on the drip irrigation adopters. For non-drip adopters, significant positive effects were observed for landholding, extension contact, and information source use. The major constraints faced by drip system adopters included a lack of knowledge about the proper operating techniques for drip systems and the cost of maintenance. CONCLUSION: The differences with inputs associated with two irrigation methods showed that the response of inputs to increase onion yield is greater for farmers who use drip irrigation than for farmers who do not, and are a result of the large differences in the technical efficiencies. These inefficiencies and other limitations following the introduction of drip irrigation, such as lack of knowledge about the proper operations, need to be addressed through tailored training for farmers and further interventions.

Potential effects of a mobile stroke unit on time to treatment and outcome in patients treated with thrombectomy or thrombolysis: A Danish-German cross-border analysis.

BACKGROUND AND PURPOSE: A mobile stroke unit (MSU) reduces delays in stroke treatment by allowing thrombolysis on board and avoiding secondary transports. Due to the beneficial effect in comparison to conventional emergency medical services, current guidelines recommend regional evaluation of MSU implementation. METHODS: In a descriptive study, current pathways of patients requiring a secondary transport for mechanical thrombectomy were reconstructed from individual patient records within a Danish (n = 122) and an adjacent German region (n = 80). Relevant timestamps included arrival times (on site, primary hospital, thrombectomy centre) as well as the initiation of acute therapy. An optimal MSU location for each region was determined. The resulting time saving was translated into averted disability-adjusted life years (DALYs). RESULTS: For each region, the optimal MSU location required a median driving time of 35 min to a stroke patient. Time savings in the German region (median [Q1; Q3]) were 7 min (-15; 31) for thrombolysis and 35 min (15; 61) for thrombectomy. In the Danish region, the corresponding time savings were 20 min (8; 30) and 43 min (25; 66). Assuming 28 thrombectomy cases and 52 thrombolysis cases this would translate to 9.4 averted DALYs per year justifying an annual net MSU budget of $0.8M purchasing power parity dollars (PPP-$) in the German region. In the Danish region, the MSU would avert 17.7 DALYs, justifying an annual net budget of PPP-$1.7M. CONCLUSION: The effects of an MSU can be calculated from individual patient pathways and reflect differences in the hospital infrastructure between Denmark and Germany.

A simple, organized web-based system improved the transfer efficiency and patient outcomes for endovascular thrombectomy in regional stroke network.

BACKGROUND: Endovascular thrombectomy (EVT) is a time-sensitive treatment for acute ischemic stroke with large vessel occlusion. To optimize transfer efficiency, a web-based platform was introduced in the Tainan Stroke Network (TSN). We assessed its application and effectiveness in regional stroke care. METHOD: This new web-based platform containing a questionnaire-style interface was introduced on October 1, 2021. To assess the transfer efficiency and patient outcomes, acute stroke patients transferred from PSCs to CSC for EVT from April 01, 2020, to December 30, 2022, were enrolled. The patients were classified into the traditional transferal pathway (TTP) group and the new transferal pathway (NTP) group depending on mode of transfer. Patient characteristics, time segments after stroke onset and outcome were compared between groups. RESULT: A total of 104 patients were enrolled, with 77 in the TTP group and 27 in the NTP group. Compared to the TTP group, the NTP group had a significantly shorter onset-to-CSC door time (TTP vs. NTP: 267 vs. 198 min; p = 0.041) and a higher EVT rate (TTP vs. NTP: 18.2% vs. 48.1%, p = 0.002). Among EVT patients, those in the NTP group had a significantly shorter CSC door-to-puncture time (TTP vs. NTP: 131.5 vs. 110 min; p = 0.029). The NTP group had a higher rate of good functional outcomes at 3 months (TTP vs. NTP: 21% vs. 61.5%; p = 0.034). CONCLUSION: This new web-based EVT transfer system provides notable improvements in clinical outcomes, transfer efficiency, and EVT execution for potential EVT candidates without markedly changing the regional stroke care paradigm.

National trends and outcomes of hospital presentation mode for endovascular stroke treatment in the U.S.

BACKGROUND: The optimal triage strategy for patients suspected of acute ischemic stroke due to large vessel occlusion (LVO) remains debated. We explored trends in presentation mode and their outcomes for mechanical thrombectomy (MT) hospitalizations based on the National Inpatient Sample (NIS) database. METHODS: We retrospectively explored the NIS database from 2016 to 2020 for stroke hospitalizations with MT. We compared outcomes at discharge for MT hospitalizations with direct vs. transferred presentation. Outcomes comprised favorable discharge disposition (home without assistance), in-hospital mortality, and radiographic intracranial hemorrhage (ICH). RESULTS: This study included 100,865 patients undergoing MT, of whom 32,685 patients (32.4 %) were transferred (median age 71[60-81] years, 16775(51.2 %) women). The utilization of MT in the U.S. nearly doubled during the study period, whereas the proportion of in-hospital transfers for MT remained unchanged (32.1-33.2 %). White race, higher presenting NIHSS, hospital size, status, and location were independent predictors of transferred status. Transferred status was significantly associated with a lower likelihood of achieving favorable outcome (OR:0.80,95 % CI: [0.72,0.89],P<0.001) and a higher likelihood of ICH (OR:1.18, 95 % CI:[1.07,1.31],P=0.001), whereas no association was observed between presentation mode and in-hospital mortality (OR:1.07,95 % CI:[0.93,1.23],P=0.33). CONCLUSION: Patients with direct presentation for MT after a stroke had better discharge outcomes and a lower risk of hemorrhagic transformation compared to those who were transferred from another facility. Determining the optimal triage strategy for MT following LVO stroke is an insightful area for future clinical trials.

Comparison between "Mothership" and "Drip and Ship" models in the management of acute ischemic strokes eligible for mechanical thrombectomy in the Charleroi area, Belgium.

BACKGROUND: In the management of acute ischemic stroke with large vessel occlusion (LVO-AIS), current data are conflicting as to whether a mothership model of management (MS) is associated with better functional recovery than a drip-and-ship model (DS). METHOD: Files from LVO-AIS patients treated with MT at CHU Charleroi were analyzed between 01/01/2017 and 12/31/2022. Consecutive patients with a LVO-AIS of the anterior circulation and a prestroke modified Rankin Scale (mRS) ≤2 were included. The study's primary endpoint was the functional independence, defined as a mRS of 0-2 at 3 months post-stroke. Times metrics of MT and thrombolysis application, safety outcome including symptomatic intracranial hemorrhage and death were recorded. We conducted similar analyses by dividing DS patients, depending of their transfer time (less or >20 min). Logistic regression was used to assess if differences in baseline characteristics affected the primary outcome. RESULTS: 366 patients were included: 229 in the DS group and 137 in the MS group. Demographic data showed a higher rate of tobacco use and lower functional status prestroke in the MS population. The MS group demonstrated better performance in time metrics related to thrombolysis and MT administration. The proportion of patients achieving an mRS of 0-2 at 3 months was similar in the DS and MS groups (50.22 % vs. 48.17 %, p = 0.706). The same conclusions were drawn from the subgroup analysis. Logistic regression analysis showed no impact of baseline characteristic differences on the primary outcome. CONCLUSION: Despite faster access to MT in the MS model, our study did not find any significant differences in functional recovery at three months post-stroke between the MS and DS management models. Our data suggest further that the prestroke health status was an important factor influencing functional outcomes after LVO-AIS.

Conservation agriculture works as a catalyst for sustainable sodic soil reclamation and enhances crop productivity and input use efficiency: A scientific inquiry.

Soil sodicity is a growing concern for crop growth and development in arid and semi-arid regions of the world. Conservation agriculture (CA) provides an effective solution towards reclamation of degraded sodic lands and enhance the crop productivity. A field experiment was carried out to assess the sodic soil reclamation potential of CA based management practices including zero tillage, legume (mungbean; Mb) rotation, residue (+R) mulch, and subsurface drip irrigation (SDI) for three years under rice-wheat (RW) system. The system scenarios (Sc) comprised of multiple indicators to measure their impact on soil properties as well as system productivity, profitability, water and nitrogen use efficiency. The results indicated that soil pHs under Sc5-Sc8 (CA-based SDI scenarios) was significantly (p < 0.05) lowered by 2.16, 2.16 and 1.33% compare with mean of Sc1 and Sc2 (CT-based system; 9.10, 8.29 and 8.14) at all three soil layers (0-5, 5-15 and 15-30 cm), respectively. Similarly, the exchangeable sodium percentage (ESP) was lowered by 2.9, 11.2 and 14.9% under CA-based scenarios with residue management compared with CT-based system (mean of Sc1 and Sc2; 15.2, 17.2 and 28.6%) during the study. The concentration of extractable anions (COЗ2‾, HCOЗ‾, Cl‾) decreased notably whereas, soil organic carbon and soil solution cations (Na+, Ca2+, Mg2+) concentration were increased under CA based management SDI plots. In addition, CA with SDI scenarios (mean of Sc5-Sc8) proved to be more productive and water-efficient than CA-based flood irrigation (FI; mean of Sc3 and Sc4). Moreover, CA-based FI and SDI scenarios saved 29.5 and 60.7% irrigation water, and improved the partial factor productivity of nitrogen (PFPN) by 6.8 and 24.4%, respectively compared to CT-R (conventional tillage without residue) based Sc1. Therefore, CA practices can potentially reduce sodicity and improve soil chemical properties for profitable crop cultivation.

Mothership versus Drip-and-Ship for stroke in a rural area: A French prospective observational study.

BACKGROUND: The availability of mechanical thrombectomy (MT) is limited. Thus, there are two paradigms for patients living closer to a primary stroke center (PSC) than a comprehensive stroke center (CSC) capable of MT: "Mothership" (direct referral to a CSC) and "Drip-and-Ship" (referral to a PSC for imaging and thrombolysis and transfer to a CSC for thrombectomy or monitoring). We aimed to compare the prognosis of patients at three months between the two paradigms in a rural area. MATERIALS: From September 2019 to March 2021, we prospectively included patients living closer to a PSC than the one CSC, regardless of the type of stroke or reperfusion treatment. The proportion of patients with a good functional outcome (Rankin≤2) at three months was compared between the two initial orientations for all patients and for subgroups: patients with ischemic stroke and patients treated by MT. RESULTS: Among the 206 patients included, 103 were admitted directly to the CSC (82.5% had an ischemic stroke and 24.3% a MT) and 103 initially admitted to a PSC and then transferred to the CSC (100% had an ischemic stroke and 52.4% a MT). The proportion of patients with a good outcome was comparable between the two groups (54.5% vs. 43.7%, P=0.22). Among the 79 patients who underwent MT, the prognosis at three months was better in the Mothership group (49.3% vs. 15.3%, P=0.01). CONCLUSION: The functional prognosis is comparable between Mothership and Drip-and-Ship paradigms in our setting, despite a trend towards a better prognosis for the Mothership. As has been shown in urban settings, the mothership paradigm also leads to a better prognosis for patients treated with MT in a rural setting.

Conventional clinical signs and symptoms are poor predictors of postoperative cerebrospinal fluid leak: A single-centre cohort perspective.

BACKGROUND: Postoperative cerebrospinal fluid (CSF) leak remains a concerning complication of the endoscopic endonasal approach (EEA) for skull base pathology. Signs and symptoms suggesting CSF leak often trigger additional workup during the postoperative course. We systematically evaluate associations between subjectively reported clinical signs/symptoms noted during the immediate postoperative period and incidence of postoperative CSF leaks. METHODS: Retrospective chart review was conducted at a tertiary academic medical centre including 137 consecutive patients with intraoperative CSF leak during EEA with primary repair between July 2018 and August 2022. Postoperative CSF leak associations with clinical signs and symptoms were evaluated using positive (PPV) and negative predictive values (NPV), sensitivity, specificity and odds ratio (OR) via univariate logistic regression. RESULTS: Seventy-nine patients (57.7%) had high-flow leaks repaired and 5 (3.6%) developed CSF leaks postoperatively. Of reported symptoms, rhinorrhea was most common (n = 52, 38.0%; PPV [95% CI] = 7.6% [4.8%, 11.9%]), followed by severe headache (n = 47, 34.3%; 6.3% [3.1%, 12.5%]), dizziness (n = 43, 31.4%; 2.3% [0.4%, 12.1%]), salty or metallic taste (n = 20, 14.6%; 9.9% [3.3%, 25.8%]), and throat drainage (n = 10, 7.3%; 9.9% [1.7%, 41.4%]). Nausea or vomiting constituted the most reported sign concerning for CSF leak (n = 73, 53.3%; PPV [95% CI] = 4.1% [2.0%, 8.1%]). On univariate regression, no sign or symptom, including rhinorrhea (OR [95% CI] = 7.00 [0.76-64.44]), throat drainage (3.42 [0.35-33.86]), salty/metallic taste (4.22 [0.66-27.04]), severe headache (3.00 [0.48-18.62]), dizziness (0.54 [0.06-4.94]), fever (3.16 [0.50-19.99]), and nausea/vomiting (1.33 [0.22-8.21]), associated with postoperative CSF leak. CONCLUSIONS: A range of subjectively reported symptoms and signs failed to predict postoperative CSF leak. Further investigation is warranted to inform appropriate attention and response.

Validation of Drug Resistance in Pneumonia (DRIP) Score as Empirical Antibiotic Failure Predictor in Community-Acquired Pneumonia Patients in Cipto Mangunkusumo Hospital.

BACKGROUND: The incidence of CAP due to Drug-Resistant Pathogen (DRP) requires broad-spectrum antibiotic therapy, Drugs Resistance in Pneumonia (DRIP) score can predict these cases. The use of the DRIP score can prevent antibiotic failure and long hospitalization, but validation is needed so that the DRIP score can be used according to the local community at Cipto Mangunkusumo National Central Public Hospital. METHODS: This research is a retrospective cohort study in CAP patients who were hospitalized during the period January 2019 to June 2020. Data were taken from medical records. Failure of empiric antibiotics occurs when one of these criteria is found: patient mortality, ICU transfer, and escalation of antibiotics as well as length of stay. RESULTS: 480 patients met the criteria. There were 331 patients (69%) with a DRIP score of <4 and 149 patients (31%) with a DRIP score of≥4. A total of 283 patients (59%) of antibiotic failures were detailed in 174 patients with a DRIP score <4 and 109 patients DRIP score ≥4. DRIP calibration using the Hosmer-Lemeshow test obtained p-value= 0.667 (p>0.05). AUC observations on the ROC curve obtained 0.651 (95% CI; 0.601-0.700). CONCLUSION: The DRIP score has low accuracy performance and calibration value in predicting empirical antibiotic failure and poor discriminatory value.

Spatiotemporal monitoring of a periodontal multispecies biofilm model: demonstration of prebiotic treatment responses.

Biofilms are complex polymicrobial communities which are often associated with human infections such as the oral disease periodontitis. Studying these complex communities under controlled conditions requires in vitro biofilm model systems that mimic the natural environment as close as possible. This study established a multispecies periodontal model in the drip flow biofilm reactor in order to mimic the continuous flow of nutrients at the air-liquid interface in the oral cavity. The design is engineered to enable real-time characterization. A community of five bacteria, Streptococcus gordonii-GFPmut3*, Streptococcus oralis-GFPmut3*, Streptococcus sanguinis-pVMCherry, Fusobacterium nucleatum, and Porphyromonas gingivalis-SNAP26 is visualized using two distinct fluorescent proteins and the SNAP-tag. The biofilm in the reactor develops into a heterogeneous, spatially uniform, dense, and metabolically active biofilm with relative cell abundances similar to those in a healthy individual. Metabolic activity, structural features, and bacterial composition of the biofilm remain stable from 3 to 6 days. As a proof of concept for our periodontal model, the 3 days developed biofilm is exposed to a prebiotic treatment with L-arginine. Multifaceted effects of L-arginine on the oral biofilm were validated by this model setup. L-arginine showed to inhibit growth and incorporation of the pathogenic species and to reduce biofilm thickness and volume. Additionally, L-arginine is metabolized by Streptococcus gordonii-GFPmut3* and Streptococcus sanguinis-pVMCherry, producing high levels of ornithine and ammonium in the biofilm. In conclusion, our drip flow reactor setup is promising in studying spatiotemporal behavior of a multispecies periodontal community.ImportancePeriodontitis is a multifactorial chronic inflammatory disease in the oral cavity associated with the accumulation of microorganisms in a biofilm. Not the presence of the biofilm as such, but changes in the microbiota (i.e., dysbiosis) drive the development of periodontitis, resulting in the destruction of tooth-supporting tissues. In this respect, novel treatment approaches focus on maintaining the health-associated homeostasis of the resident oral microbiota. To get insight in dynamic biofilm responses, our research presents the establishment of a periodontal biofilm model including Streptococcus gordonii, Streptococcus oralis, Streptococcus sanguinis, Fusobacterium nucleatum, and Porphyromonas gingivalis. The added value of the model setup is the combination of simulating continuously changing natural mouth conditions with spatiotemporal biofilm profiling using non-destructive characterization tools. These applications are limited for periodontal biofilm research and would contribute in understanding treatment mechanisms, short- or long-term exposure effects, the adaptation potential of the biofilm and thus treatment strategies.

The effect of drip versus intermittent feeding on splanchnic oxygenation in preterm infants with intrauterine growth restriction: a prospective randomized trial.

The main purpose of this study was to evaluate the impact of drip versus intermittent feeding on splanchnic oxygenation in preterm infants with intrauterine growth restriction. The second objective was to assess the relationship between fetal splanchnic circulation parameters and splanchnic oxygenation during the first week of life. A single-center, prospective, randomized study with 51 fetuses/infants was conducted. Fetal Doppler measurements including umbilical artery, middle cerebral artery, and superior mesenteric artery (SMA) were recorded in IUGR fetuses. After preterm delivery, the infants were randomly assigned to one of two feeding modalities: drip (3-h continuous) or intermittent (bolus in 10 min). Continuous regional splanchnic saturation (rSO2S) monitoring was carried out during the first week of life, simultaneously with continuous oxygen arterial saturation (SaO2) monitoring, and the infants' fractional oxygen extractions (FOE) were calculated. These parameters were evaluated as means on a daily basis for the first week of life, as well as pre-prandial and post-prandial measurements on the seventh day. Fetal Doppler flow velocimetry disturbances were present in 72.5% of the study cohort. The drip (26 infants) and intermittent (25 infants) groups were similar in demographic and clinical characteristics, as well as the prevalence of feeding intolerance and necrotizing enterocolitis. During the first week of life, there was no difference in daily mean rSO2S and FOE values between the drip and intermittent groups, whereas unfed infants had mostly lower rSO2S values. Pre-prandial and post-prandial rSO2S values remained stable in both groups. Also, no association was detected between fetal splanchnic circulation parameters and neonatal splanchnic oxygenation. RSO2S values were strongly correlated to gestational age and birth weight. During the whole week, except for the first 2 days, infants with umbilical catheters had significantly lower rSO2S values than infants without.  Conclusion: Our data suggest that the key factor in splanchnic oxygenation is feeding, not the feeding modality. In addition, the umbilical vein catheter had a negative impact on splanchnic oxygenation.  Clinical Trial Registration: The Effect of Neonatal Feeding Modalities on Splanchnic Oxygenation, NCT05513495,  https://clinicaltrials.gov/ct2/results cond=&term=NCT05513495&cntry=TR&state=&city=&dist= . Retrospectively registered, date of registration: August 2022. What is Known: • It is known that preterm infants with IUGR are at increased risk of hypoxic-ischemic intestinal damage and impaired splanchnic oxygenation. What is New: • The key factor in splanchnic oxygenation of preterm infants with IUGR is feeding, not the feeding modality (drip or intermittent). • In addition, the umbilical vein catheter had a negative impact on splanchnic oxygenation.

Cold-tolerant microorganisms causing spoilage of vacuum-packed beef under time-temperature abuse determined by culture and qPCR.

Vacuum packaging and storage conditions at chilled temperatures are commonly used in order to prolong the shelf life of meat. Under these conditions and time-temperature abuse, cold-tolerant (facultatively) anaerobic spoilage microorganisms can continue growing. This study investigated growth of six relevant spoilage microorganisms in vacuum-packed beef (n = 12, 72 subsamples, stored at 10 °C for 28 days) using culture and qPCR methods. Correspondingly, six qPCRs were newly developed/modified (for total bacteria, lactic acid bacteria (LAB), Enterobacterales, total fungi, Kazachstania psychrophila, and cold-tolerant Clostridium spp.). Besides microbial quantification, four spoilage appearances of meat (gas production, spoilage odor, % drip loss, and meat color) were observed. Results obtained from culture and qPCR show that total bacteria, LAB, and Enterobacterales reached their stationary phase at day 7 when spoilage parameters such as gas production were statistically increased and a deviation of odor was detected. Fastidious cold-tolerant Clostridium spp. and K. psychrophila could be detected from day 7. Based on microbiological and sensory analysis results, the maximum shelf life of vacuum-packed beef stored at 10 °C is 7 days. The developed qPCR has the potential to be used as an alternative method to culturing for determination of microbial growth.

Apex structures enhance water drainage on leaves.

The rapid removal of rain droplets at the leaf apex is critical for leaves to avoid damage under rainfall conditions, but the general water drainage principle remains unclear. We demonstrate that the apex structure enhances water drainage on the leaf by employing a curvature-controlled mechanism that is based on shaping a balance between reduced capillarity and enhanced gravity components. The leaf apex shape changes from round to triangle to acuminate, and the leaf surface changes from flat to bent, resulting in the increase of the water drainage rate, high-dripping frequencies, and the reduction of retention volumes. For wet tropical plants, such as Alocasia macrorrhiza, Gaussian curvature reconfiguration at the drip tip leads to the capillarity transition from resistance to actuation, further enhancing water drainage to the largest degree possible. The phenomenon is distinct from the widely researched liquid motion control mechanisms, and it offers a specific parametric approach that can be applied to achieve the desired fluidic behavior in a well-controlled way.

Kinetically distinct processing pathways diversify the CD8+ T cell response to a single viral epitope.

The source proteins from which CD8+ T cell-activating peptides are derived remain enigmatic. Glycoproteins are particularly challenging in this regard owing to several potential trafficking routes within the cell. By engineering a glycoprotein-derived epitope to contain an N-linked glycosylation site, we determined that optimal CD8+ T cell expansion and function were induced by the peptides that are rapidly produced from the exceedingly minor fraction of protein mislocalized to the cytosol. In contrast, peptides derived from the much larger fraction that undergoes translocation and quality control are produced with delayed kinetics and induce suboptimal CD8+ T cell responses. This dual system of peptide generation enhances CD8+ T cell participation in diversifying both antigenicity and the kinetics of peptide display.

Generalizing routes of plant exposure to pesticides by plant uptake models to assess pesticide application efficiency.

Pesticide application techniques are critical not only for integrated pest management (IPM) but also for food and environmental safety. Assessing pesticide application efficiency on plants can help optimize IPM and reduce pesticide environmental impacts. With hundreds of pesticides registered for use in agriculture, this study proposed a modeling approach based on plant uptake models for generalizing routes of plant chemical exposures that can correspond to different types of pesticide application methods and evaluating their respective efficiency on plants. Three representative pesticide application methods (i.e., drip irrigation, foliar spray, and broadcast application) were selected for modeling simulations. The simulation results for three representative pesticides (i.e., halofenozide, pymetrozine, and paraquat) revealed that the soil-based transpiration exposure route facilitated the bioaccumulation of moderately lipophilic compounds in leaves and fruits. While the plant surface-based exposure route (i.e., leaf cuticle penetration) made it easier for highly lipophilic compounds to enter plants, moderately lipophilic pesticides (i.e., log KOW ∼ 2) were more soluble in phloem sap, which enhanced their subsequent transport within plant tissues. In general, moderately lipophilic pesticides had the highest simulated residue concentrations in plant tissues for the three specific application methods, indicating they had the highest application efficiency due to their enhanced uptake routes (via transpiration and surface penetration) and increased solubility in xylem and phloem saps. Compared to foliar spray and broadcast application, drip irrigation produced higher residue concentrations for a wide variety of pesticides, exhibiting the highest application efficiency for many pesticides, especially for moderately lipophilic compounds. Future research should incorporate plant growth stages, crop safety, pesticide formulations, and multiple application events into the modeling approach for understanding pesticide application efficiency evaluation.

Influence of microclimate control on the growth of asparagus under greenhouse in tropical climates.

High temperatures have become common in cities in Taiwan, and this phenomenon has spread to surrounding agricultural areas. Tainan, a city located in a tropical climate zone with agriculture as its primary development industry, is one of the cities considerably affected by the high temperature. High temperatures can reduce crop yields and even cause plant death, especially for vulnerable high-value crops, which are severely to microclimate conditions. Asparagus is a high-value crop that has long been cultivated in the Jiangjun District of Tainan. Recently, asparagus has been planted in greenhouses to protect against pests and natural disasters. However, the greenhouses can overheat. To identify the optimal growth environment for asparagus, this study applies vertical monitoring to record the temperature in the greenhouse and the soil moisture content of a control (canal irrigation) and an experimental (drip irrigation) group. When the surface layer of the soil exceeds 33°C, the tender stems of asparagus bloom readily, reducing its commercial value. Therefore, drip irrigation was conducted with cool water (26°C) to reduce soil temperature in summer and warm water (28°C) to increase soil temperature in winter. The study also recorded the growth of asparagus using daily yields measured by farmers during weighing and packing to understand the benefits of controlling the greenhouse microclimate. This study reports a correlation of 0.85 between asparagus yield and temperature and a correlation of 0.86 between asparagus yield and soil moisture content. The use of a drip irrigation system with a water temperature adjustment function not only saves up to 50% of water but also resulted in an average yield increase of 10% through maintaining stable soil moisture content and temperature. Therefore, the findings of this study can be applied to asparagus yields affected by high temperature and can solve the problems of poor quality in summer and low yield in winter.

Subcutaneous Insulin Versus Traditional Intravenous Insulin Infusion in Treatment of Mild to Moderate Diabetic Ketoacidosis.

BACKGROUND: Intravenous (IV) insulin infusions are the current standard of care for treatment of diabetic ketoacidosis (DKA). Subcutaneous (SQ) insulin, however, may also be a safe and effective alternative. OBJECTIVE: The purpose of this study was to compare patient-centered outcomes related to the treatment of mild to moderate DKA using two different protocols: an SQ insulin protocol and an IV insulin infusion protocol with an initial bolus (IVB) or without a bolus (IVNB). METHODS: We retrospectively conducted a multicenter cohort study evaluating SQ vs. IV insulin for the treatment of mild to moderate DKA. The primary outcome was time to DKA resolution. Secondary outcomes included time to glucose correction, hospital length of stay (LOS), intensive care unit LOS, hypoglycemia events, readmission rates, and IV insulin use. RESULTS: Within the study time frame, 257 patients were included in the multivariate Cox proportional hazards regression analysis. There was no significant difference in the time to DKA resolution between the IVB (p = 0.603) or IVNB (p = 0.269) groups compared with the population who received SQ insulin only. Hospital LOS was significantly longer when comparing the SQ group with the IVNB group (p < 0.001), but not when comparing it with the IVB group (p = 0.259). The IV protocols had significantly more hypoglycemic events compared with the SQ protocol (IVB vs. SQ, p < 0.001; IVNB vs. SQ, p = 0.001). CONCLUSIONS: SQ insulin may be an effective alternative option for treating mild to moderate DKA with fewer hypoglycemic effects.