Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

A multimodal cell census and atlas of the mammalian primary motor cortex

ABSTRACT We report the generation of a multimodal cell census and atlas of the mammalian primary motor cortex (MOp or M1) as the initial product of the BRAIN Initiative Cell Census Network (BICCN). This was achieved by coordinated large-scale analyses of single-cell transcriptomes, chromatin accessibility, DNA methylomes, spatially resolved single-cell transcriptomes, morphological and electrophysiological properties, and cellular resolution input-output mapping, integrated through cross-modal computational analysis. Together, our results advance the collective knowledge and understanding of brain cell type organization: First, our study reveals a unified molecular genetic landscape of cortical cell types that congruently integrates their transcriptome, open chromatin and DNA methylation maps. Second, cross-species analysis achieves a unified taxonomy of transcriptomic types and their hierarchical organization that are conserved from mouse to marmoset and human. Third, cross-modal analysis provides compelling evidence for the epigenomic, transcriptomic, and gene regulatory basis of neuronal phenotypes such as their physiological and anatomical properties, demonstrating the biological validity and genomic underpinning of neuron types and subtypes. Fourth, in situ single-cell transcriptomics provides a spatially-resolved cell type atlas of the motor cortex. Fifth, integrated transcriptomic, epigenomic and anatomical analyses reveal the correspondence between neural circuits and transcriptomic cell types. We further present an extensive genetic toolset for targeting and fate mapping glutamatergic projection neuron types toward linking their developmental trajectory to their circuit function. Together, our results establish a unified and mechanistic framework of neuronal cell type organization that integrates multi-layered molecular genetic and spatial information with multi-faceted phenotypic properties

Burden of Healthcare Utilization among Chronic Obstructive Pulmonary Disease Patients with and without Cancer Receiving Palliative Care: A Population-Based Study in Taiwan

Chronic obstructive pulmonary disease (COPD) is a chronic disease that burdens patients worldwide. This study aims to discover the burdens of health services among COPD patients who received palliative care (PC). Study subjects were identified as COPD patients with ICU and PC records between 2009 and 2013 in Taiwan’s National Health Insurance Research Database. The burdens of healthcare utilization were analyzed using logistic regression to estimate the difference between those with and without cancer. Of all 1215 COPD patients receiving PC, patients without cancer were older and had more comorbidities, higher rates of ICU admissions, and longer ICU stays than those with cancer. COPD patients with cancer received significantly more blood transfusions (Odds Ratio, OR: 1.66; 95% C.I.: 1.11–2.49) and computed tomography scans (OR: 1.88; 95% C.I.: 1.10–3.22) compared with those without cancer. Bronchoscopic interventions (OR: 0.26; 95% C.I.: 0.07–0.97) and inpatient physical restraints (OR: 0.24; 95% C.I.: 0.08–0.72) were significantly more utilized in patients without cancer. COPD patients without cancer appeared to receive more invasive healthcare interventions than those without cancer. The unmet needs and preferences of patients in the life-limiting stage should be taken into consideration for the quality of care in the ICU environment

Structure of the malaria vaccine candidate Pfs48/45 and its recognition by transmission blocking antibodies

An effective malaria vaccine remains a global health priority and vaccine immunogens which prevent transmission of the parasite will have important roles in multi-component vaccines. One of the most promising candidates for inclusion in a transmission-blocking malaria vaccine is the gamete surface protein Pfs48/45, which is essential for development of the parasite in the mosquito midgut. Indeed, antibodies which bind Pfs48/45 can prevent transmission if ingested with the parasite as part of the mosquito bloodmeal. Here we present the structure of full-length Pfs48/45, showing its three domains to form a dynamic, planar, triangular arrangement. We reveal where transmission-blocking and non-blocking antibodies bind on Pfs48/45. Finally, we demonstrate that antibodies which bind across this molecule can be transmission-blocking. These studies will guide the development of future Pfs48/45-based vaccine immunogens

Biventricular myocardial adaptation in patients with repaired tetralogy of Fallot: Mechanistic insights from magnetic resonance imaging tissue phase mapping.

BACKGROUND:The myocardial adaptive mechanism in patients with repaired tetralogy of Fallot (rTOF) is less understood. We aimed to investigate biventricular myocardial adaptive remodeling in rTOF patients. METHODS:We recruited 32 rTOF patients and 38 age- and sex-matched normal controls. The pulmonary stenosis of rTOF patients was measured using catheterized pressure gradient between right ventricle (RV) and pulmonary artery (PGRVPA). rTOF patients with PGRVPA < 15 mmHg and ≥15 mmHg were classified as low pulmonary stenosis (rTOFlow, n = 19) and high pulmonary stenosis (rTOFhigh, n = 13) subgroups, respectively. Magnetic resonance imaging tissue phase mapping was employed to evaluate the voxelwise biventricular myocardial motion in longitudinal (Vz), radial (Vr), and circumferential (Vφ) directions. RESULTS:The rTOFlow subgroup presented higher pulmonary regurgitation fraction than rTOFhigh subgroup (p < 0.001). Compared with the normal group, only rTOFlow subgroup presented a decreased RV ejection fraction (RVEF) (p < 0.05). The rTOFlow subgroup showed decreased systolic and diastolic Vz in RV and LV, whereas rTOFhigh subgroup showed such change only in RV. In rTOFlow subgroup, RVEF significantly correlated with RV systolic Vr (r = 0.56, p < 0.05), whereas LVEF correlated with LV systolic Vz (r = 0.51, p = 0.02). Prolonged QRS correlated with RV systolic Vr (r = -0.58, p < 0.01) and LV diastolic Vr (r = 0.81, p < 0.001). No such correlations occurred in rTOFhigh subgroup. CONCLUSIONS:The avoidance of unfavorable functional interaction in RV and LV in rTOFhigh subgroup suggested that adequate pulmonary stenosis (PGRVPA ≥ 15 mmHg in this sereis) has a protective effect against pulmonary regurgitation

Age-related Differences in the Clinical Presentation, Associated Metabolic Abnormality, and Estimated Cardiovascular Risks from Nonalcoholic Fatty Liver Disease: A Cross-sectional Study from Health Evaluation Center in Taiwan

Background: Nonalcoholic fatty liver disease (NAFLD), which represents metabolic abnormality and reflects extra visceral fat deposition, has been shown to be a threat to public health and associated with cardiovascular risks. However, data regarding the differences in clinical presentation and related metabolic abnormalities in the aging group population remained scarce in the Taiwanese population. Methods: We subsequently examined 8,658 subjects participating general health evaluation in Mackay Memorial hospital from 2003 to 2007. Baseline characteristics, anthropometrics, medical history, and biochemical markers were all collected. Abdominal ultrasonography was performed in all subjects. Two estimated cardiovascular risk scores were calculated according to criteria of the U.S. National Cholesterol Education Program Adult Treatment Panel III as metabolic score and Framingham risk score. Univariate logistic regression model was used to examine whether the baseline characteristics, anthropometrics, histories, and biochemical markers were independently associated with NAFLD from various age groups (young vs. aging groups) classified by 60 years of age. Respective receiver operating characteristic curves (ROC) with area under the curve were generated to test the capability of both cardiovascular risk scores in NAFLD discrimination from different age groups. Results: Totally 7,204 subjects (mean age: 44.5±11 years, 36% female) were finally enrolled in our study. Subjects with NAFLD were observed to have high body weight, body mass index, and circumferential waist and significantly abnormal biochemical markers accompanying worsening lipid profiles when compared with those without NAFLD in both the young and aging group populations (all p<0.001) although alkaline phosphatase, total cholesterol, and low-density lipoprotein did not show significant differences with and without NAFLD in the aging group population. Diabetes history remained a strong independent predictor of NAFLD in both young and aging groups (odds ratio: 3.3, p<0.001 vs. 2.51, p=0.014). The prediction model by using different cardiovascular risk scores yielded a meaningful ROC value of 0.82 (young group) and 0.71 (aging group) (both p<0.001) for metabolic score with ROC value of 0.67 (young group) and 0.52 (aging group) for Framingham risk score (p<0.001 vs. 0.408, respectively). Conclusion: The prevalence of NAFLD demonstrated a bimodal distribution with age in different genders. Although baseline characters and biochemical markers were demonstrated to be potential screening tools in detecting such clinical abnormality, they actually exerted diverse capabilities in the prediction of NAFLD in the different age groups. Traditional cardiovascular risk scores were less effective in predicting NAFLD in the aging group population

Influence of vaccination on critical COVID-19 patients with acute respiratory failure: a retrospective cohort study

Abstract Background Despite vaccines’ effectiveness in reducing COVID-19 infection rates and disease severity, their impact on critical patients presenting with acute respiratory failure is elusive. The aim of this study was to further investigate the influence of vaccination on mortality rates among severely ill COVID-19 patients experiencing acute respiratory failure. Methods This retrospective cohort study was carried out at a tertiary medical center in Taiwan. From April to September 2022, patients who tested positive for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) through reverse transcription polymerase chain reaction (RT-PCR) and subsequently experienced acute respiratory failure were included in the study. Baseline characteristics, including vaccination history, along with information regarding critical illness and clinical outcomes, were gathered and compared between patients who received the vaccine and those who did not. Results A total of 215 patients with COVID-19 exhibiting acute respiratory failure, as confirmed via RT‒PCR, were included in the analysis. Of this cohort, sixty-six (30.7%) patients died within 28 days. Neither administration of the vaccine nor achievement of primary series vaccination status had a significantly different effect on 28 day mortality, number of viral shedding events, acute respiratory distress syndrome (ARDS) incidence or other clinical outcomes. Patients who received the booster vaccine and completed the primary series showed a tendency of increased 28 days of ventilator-free status, though this difference was not statistically significant (p = 0.815). Conclusions Vaccination status did not significantly influence mortality rates, the occurrence of ARDS, or the viral shedding duration in COVID-19 patients with acute respiratory failure

High-Efficiency Broadband Anomalous Reflection by Gradient Meta-Surfaces

We combine theory and experiment to demonstrate that a carefully designed gradient meta-surface supports high-efficiency anomalous reflections for near-infrared light following the generalized Snell’s law, and the reflected wave becomes a bounded surface wave as the incident angle exceeds a critical value. Compared to previously fabricated gradient meta-surfaces in infrared regime, our samples work in a shorter wavelength regime with a broad bandwidth (750–900 nm), exhibit a much higher conversion efficiency (∼80%) to the anomalous reflection mode at normal incidence, and keep light polarization unchanged after the anomalous reflection. Finite-difference-time-domain (FDTD) simulations are in excellent agreement with experiments. Our findings may lead to many interesting applications, such as antireflection coating, polarization and spectral beam splitters, high-efficiency light absorbers, and surface plasmon couplers