HRP-OG: Online Learning with Generative Feature Replay for Hypertension Risk Prediction in a Nonstationary Environment.

Hypertension is a major risk factor for many serious diseases. With the aging population and lifestyle changes, the incidence of hypertension continues to rise, imposing a significant medical cost burden on patients and severely affecting their quality of life. Early intervention can greatly reduce the prevalence of hypertension. Research on hypertension early warning models based on electronic health records (EHRs) is an important and effective method for achieving early hypertension warning. However, limited by the scarcity and imbalance of multivisit records, and the nonstationary characteristics of hypertension features, it is difficult to predict the probability of hypertension prevalence in a patient effectively. Therefore, this study proposes an online hypertension monitoring model (HRP-OG) based on reinforcement learning and generative feature replay. It transforms the hypertension prediction problem into a sequential decision problem, achieving risk prediction of hypertension for patients using multivisit records. Sensors embedded in medical devices and wearables continuously capture real-time physiological data such as blood pressure, heart rate, and activity levels, which are integrated into the EHR. The fit between the samples generated by the generator and the real visit data is evaluated using maximum likelihood estimation, which can reduce the adversarial discrepancy between the feature space of hypertension and incoming incremental data, and the model is updated online based on real-time data using generative feature replay. The incorporation of sensor data ensures that the model adapts dynamically to changes in the condition of patients, facilitating timely interventions. In this study, the publicly available MIMIC-III data are used for validation, and the experimental results demonstrate that compared to existing advanced methods, HRP-OG can effectively improve the accuracy of hypertension risk prediction for few-shot multivisit record in nonstationary environments.

Exploration and future trends on spatial correlation of green innovation efficiency in strategic emerging industries under the digital economy: A social network analysis.

With digital technological change and the increasing frequency of interregional innovation links, the spatial correlation and diversity of strategic emerging industries' green innovation efficiency (SEI-GIE) need to be explored in depth. This paper innovatively constructs the SEI-GIE input-output index system under digital economy. The proposed grey model FINGBM(1,1) with ω-order accumulation and weighted initial value optimization realizes effective prediction of 7 input-output indicators of 30 provinces in China from 2021 to 2025. Super-SBM-DEA, gravity model, and social network analysis are applied to explore spatial network structure's dynamic process of SEI-GIE from 12th to 14th Five-Year-Plan period (2011-2025). Empirical results show that (1) Under the effect of digital economy, the SEI-GIE in China generally shows a U-shaped fluctuation trend, in which the growth trend in the central region is obvious, and the western region shows significant fluctuations. (2) The spatial correlation network of SEI-GIE presents a complex and stable center-periphery circle. Particularly, the overall increase in network efficiency highlights the strong small-world characteristics. (3) Beijing, Shanghai, Zhejiang and Jiangsu have always been in the leading core position, with strong influence and control; And Tianjin's core position in the network will decline. Additionally, Guangxi and Chongqing have great potential, but Guangdong needs to strengthen its radiation effect. (4) Block model shows that plate-I (Beijing, Tianjin) receive spatial spillovers from others, while plates-III,IV have significant spillover effects. This study provides theoretical reference for policymakers from a network perspective to promote development of China's SEI-GIE.

Generation of sheep with defined FecBB and TBXT mutations and porcine blastocysts with KCNJ5G151R/+ mutation using prime editing.

BACKGROUND: Rewriting the genomes of living organisms has been a long-standing aim in the biological sciences. The revelation of the CRISPR/Cas9 technology has revolutionized the entire biological field. Since its emergence, this technology has been widely applied to induce gene knockouts, insertions, deletions, and base substitutions. However, the classical version of this system was imperfect for inducing or correcting desired mutations. A subsequent development generated more advanced classes, including cytosine and adenine base editors, which can be used to achieve single nucleotide substitutions. Nevertheless, these advanced systems still suffer from several limitations, such as the inability to edit loci without a suitable PAM sequence and to induce base transversions. On the other hand, the recently emerged prime editors (PEs) can achieve all possible single nucleotide substitutions as well as targeted insertions and deletions, which show promising potential to alter and correct the genomes of various organisms. Of note, the application of PE to edit livestock genomes has not been reported yet. RESULTS: In this study, using PE, we successfully generated sheep with two agriculturally significant mutations, including the fecundity-related FecBB p.Q249R and the tail length-related TBXT p.G112W. Additionally, we applied PE to generate porcine blastocysts with a biomedically relevant point mutation (KCNJ5 p.G151R) as a porcine model of human primary aldosteronism. CONCLUSIONS: Our study demonstrates the potential of the PE system to edit the genomes of large animals for the induction of economically desired mutations and for modeling human diseases. Although prime-edited sheep and porcine blastocysts could be generated, the editing frequencies are still unsatisfactory, highlighting the need for optimizations in the PE system for efficient generation of large animals with customized traits.

Markhor-derived Introgression of a Genomic Region Encompassing PAPSS2 Confers High-altitude Adaptability in Tibetan Goats.

Understanding the genetic mechanism of how animals adapt to extreme conditions is fundamental to determine the relationship between molecular evolution and changing environments. Goat is one of the first domesticated species and has evolved rapidly to adapt to diverse environments, including harsh high-altitude conditions with low temperature and poor oxygen supply but strong ultraviolet radiation. Here, we analyzed 331 genomes of domestic goats and wild caprid species living at varying altitudes (high > 3000 m above sea level and low < 1200 m), along with a reference-guided chromosome-scale assembly (contig-N50: 90.4 Mb) of a female Tibetan goat genome based on PacBio HiFi long reads, to dissect the genetic determinants underlying their adaptation to harsh conditions on the Qinghai-Tibetan Plateau (QTP). Population genomic analyses combined with genome-wide association studies (GWAS) revealed a genomic region harboring the 3'-phosphoadenosine 5'-phosphosulfate synthase 2 (PAPSS2) gene showing strong association with high-altitude adaptability (PGWAS = 3.62 × 10-25) in Tibetan goats. Transcriptomic data from 13 tissues revealed that PAPSS2 was implicated in hypoxia-related pathways in Tibetan goats. We further verified potential functional role of PAPSS2 in response to hypoxia in PAPSS2-deficient cells. Introgression analyses suggested that the PAPSS2 haplotype conferring the high-altitude adaptability in Tibetan goats originated from a recent hybridization between goats and a wild caprid species, the markhor (Capra falconeri). In conclusion, our results uncover a hitherto unknown contribution of PAPSS2 to high-altitude adaptability in Tibetan goats on QTP, following interspecific introgression and natural selection.

External counterpulsation stimulation combined with acupuncture for vascular endothelial function in patients with hypertension: A randomized pilot trial.

OBJECTIVE: Endothelial function may improve with enhanced external counterpulsation (EECP) or acupuncture. This study aimed to evaluate the feasibility of acupoint stimulation combined with EECP (acupoint-EECP) for endothelial cell function in patients with essential hypertension. METHODS: Thirty essential hypertensive patients were, randomly divided into two groups, with 15 patients in the acupoint-EECP group, and 15 patients in the control group, of which 3 cases were lost by week 6. Both groups were treated with continued medicine. The participants in the acupoint-EECP group received acupoint stimulation combined with EECP therapy, 45 min for each time, 5 times weekly for 6 weeks for a total of 22.5 hours. The selected acupoints are Zusanli (ST36), Fenglong (ST40) and Sanyinjiao (SP6). The curative effects of the two groups were compared. RESULTS: The acupoint-EECP group (n=15) showed significant improvement in endothelial function [nitric oxide (NO) ，endothelin-1 (ET-1) and carotid-femoral pulse wave velocity (cf-PWV), respectively] values compared to the control group (n=12). Multiple imputation (n = 20 imputations) was performed to account for potential bias due to missing data. In stratified analyses, SBP and DBP values decreased when the baseline SBP was ≥120 mmHg and DBP was ≥80 mmHg. CONCLUSIONS: These findings suggest the feasibility of acupoint-EECP in improving endothelial function and treating hypertension. (The Chinese clinical trial registration number is ChiCTR2100053795.).

Identification of Driver Status Hazard Level and the System.

According to the survey statistics, most traffic accidents are caused by the driver's behavior and status irregularities. Because there is no multi-level dangerous state grading system at home and abroad, this paper proposes a complex state grading system for real-time detection and dynamic tracking of the driver's state. The system uses OpenMV as the acquisition camera combined with the cradle head tracking system to collect the driver's current driving image in real-time dynamically, combines the YOLOX algorithm with the OpenPose algorithm to judge the driver's dangerous driving behavior by detecting unsafe objects in the cab and the driver's posture, and combines the improved Retinaface face detection algorithm with the Dlib feature-point algorithm to discriminate the fatigue driving state of the driver. The experimental results show that the accuracy of the three driver danger levels (R1, R2, and R3) obtained by the proposed system reaches 95.8%, 94.5%, and 96.3%, respectively. The experimental results of this system have a specific practical significance in driver-distracted driving warnings.

Optimized Cas9:sgRNA delivery efficiently generates biallelic MSTN knockout sheep without affecting meat quality.

BACKGROUND: CRISPR/Cas9-based genome-editing systems have been used to efficiently engineer livestock species with precise genetic alterations intended for biomedical and agricultural applications. Previously, we have successfully generated gene-edited sheep and goats via one-cell-stage embryonic microinjection of a Cas9 mRNA and single-guide RNAs (sgRNAs) mixture. However, most gene-edited animals produced using this approach were heterozygotes. Additionally, non-homozygous gene-editing outcomes may not fully generate the desired phenotype in an efficient manner. RESULTS: We report the optimization of a Cas9 mRNA-sgRNA delivery system to efficiently generate homozygous myostatin (MSTN) knockout sheep for improved growth and meat production. Firstly, an sgRNA selection software (sgRNAcas9) was used to preliminarily screen for highly efficient sgRNAs. Ten sgRNAs targeting the MSTN gene were selected and validated in vitro using sheep fibroblast cells. Four out of ten sgRNAs (two in exon 1 and two in exon 2) showed a targeting efficiency > 50%. To determine the optimal CRISPR/Cas9 microinjection concentration, four levels of Cas9 mRNA and three levels of sgRNAs in mixtures were injected into sheep embryos. Microinjection of 100 ng/µL Cas9 mRNA and 200 ng/µL sgRNAs resulted in the most improved targeting efficiency. Additionally, using both the highly efficient sgRNAs and the optimal microinjection concentration, MSTN-knockout sheep were generated with approximately 50% targeting efficiency, reaching a homozygous knockout efficiency of 25%. Growth rate and meat quality of MSTN-edited lambs were also investigated. MSTN-knockout lambs exhibited increased body weight and average daily gain. Moreover, pH, drip loss, intramuscular fat, crude protein, and shear force of gluteal muscles of MSTN-knockout lambs did not show changes compared to the wild-type lambs. CONCLUSIONS: This study highlights the importance of in vitro evaluation for the optimization of sgRNAs and microinjection dosage of gene editing reagents. This approach enabled efficient engineering of homozygous knockout sheep. Additionally, this study confirms that MSTN-knockout lambs does not negatively impact meat quality, thus supporting the adoption of gene editing as tool to improve productivity of farm animals.

Inactivation of the MSTN gene expression changes the composition and function of the gut microbiome in sheep.

BACKGROUND: Myostatin (MSTN) negatively regulates the muscle growth in animals and MSTN deficient sheep have been widely reported previously. The goal of this study was to explore how MSTN inactivation influences their gut microbiota composition and potential functions. RESULTS: We compared the slaughter parameters and meat quality of 3 MSTN-edited male sheep and 3 wild-type male sheep, and analyzed the gut microbiome of the MSTN-edited sheep (8 female and 8 male sheep) and wild-type sheep (8 female and 8 male sheep) through metagenomic sequencing. The results showed that the body weight, carcass weight and eye muscle area of MSTN-edited sheep were significantly higher, but there were no significant differences in the meat quality indexes. At the microbial level, the alpha diversity was significantly higher in the MSTN-edited sheep (P < 0.05), and the microbial composition was significantly different by PCoA analysis in the MSTN-edited and wild-type sheep. The abundance of Firmicutes significantly increased and Bacteroidota significantly decreased in the MSTN-edited sheep. At genus level, the abundance of Flavonifractor, Subdoligranulum, Ruthenibacterium, Agathobaculum, Anaerotignum, Oribacterium and Lactobacillus were significantly increased in the MSTN-edited sheep (P < 0.05). Further analysis of functional differences was found that the carotenoid biosynthesis was significantly increased and the peroxisome, apoptosis, ferroptosis, N-glycan biosynthesis, thermogenesis, and adipocytokines pathways were decreased in the MSTN-edited sheep (P < 0.05). Moreover, carbohydrate-active enzymes (CAZymes) results certified the abundance of the GH13_39, GH4, GH137, GH71 and PL17 were upregulated, and the GT41 and CBM20 were downregulated in the MSTN-edited sheep (P < 0.05). CONCLUSIONS: Our study suggested that MSTN inactivation remarkably influenced the composition and potential function of hindgut microbial communities of the sheep, and significantly promoted growth performance without affecting meat quality.

Transcriptional Regulation of Solute Carrier (SLC) Drug Transporters.

Facilitated transport is necessitated for large size, charged, and/or hydrophilic drugs to move across the membrane. The drug transporters in the solute carrier (SLC) superfamily, mainly including organic anion-transporting polypeptides (OATPs), organic anion transporters (OATs), organic cation transporters (OCTs), organic cation/carnitine transporters (OCTNs), peptide transporters (PEPTs), and multidrug and toxin extrusion proteins (MATEs), are critical facilitators of drug transport and distribution in human body. The expression of these SLC drug transporters is found in tissues throughout the body, with high abundance in the epithelial cells of major organs for drug disposition, such as intestine, liver, and kidney. These SLC drug transporters are clinically important in drug absorption, metabolism, distribution, and excretion. The mechanisms underlying their regulation have been revealing in recent years. Epigenetic and nuclear receptor-mediated transcriptional regulation of SLC drug transporters have particularly attracted much attention. This review focuses on the transcriptional regulation of major SLC drug transporter genes. Revealing the mechanisms underlying the transcription of those critical drug transporters will help us understand pharmacokinetics and pharmacodynamics, ultimately improving drug therapeutic effectiveness while minimizing drug toxicity. Significance Statement It has become increasingly recognized that solute carrier (SLC) drug transporters play a crucial, and sometimes determinative, role in drug disposition and response, which is reflected in decision-making during not only clinical drug therapy but also drug development. Understanding the mechanisms accounting for the transcription of these transporters is critical to interpret their abundance in various tissues under different conditions, which is necessary to clarify the pharmacological response, adverse effects, and drug-drug interactions for clinically used drugs.

Genetics of the phenotypic evolution in sheep: a molecular look at diversity-driving genes.

BACKGROUND: After domestication, the evolution of phenotypically-varied sheep breeds has generated rich biodiversity. This wide phenotypic variation arises as a result of hidden genomic changes that range from a single nucleotide to several thousands of nucleotides. Thus, it is of interest and significance to reveal and understand the genomic changes underlying the phenotypic variation of sheep breeds in order to drive selection towards economically important traits. REVIEW: Various traits contribute to the emergence of variation in sheep phenotypic characteristics, including coat color, horns, tail, wool, ears, udder, vertebrae, among others. The genes that determine most of these phenotypic traits have been investigated, which has generated knowledge regarding the genetic determinism of several agriculturally-relevant traits in sheep. In this review, we discuss the genomic knowledge that has emerged in the past few decades regarding the phenotypic traits in sheep, and our ultimate aim is to encourage its practical application in sheep breeding. In addition, in order to expand the current understanding of the sheep genome, we shed light on research gaps that require further investigation. CONCLUSIONS: Although significant research efforts have been conducted in the past few decades, several aspects of the sheep genome remain unexplored. For the full utilization of the current knowledge of the sheep genome, a wide practical application is still required in order to boost sheep productive performance and contribute to the generation of improved sheep breeds. The accumulated knowledge on the sheep genome will help advance and strengthen sheep breeding programs to face future challenges in the sector, such as climate change, global human population growth, and the increasing demand for products of animal origin.

Deletions in the KAP6-1 gene are associated with fiber traits in cashmere-producing goats.

Keratin-associated proteins (KAPs) are important structural components of fibers that predominantly present in the ortho-cortex. These proteins form a cross-linked network with keratin intermediate filaments (KIFs), thus producing a strong hair shaft. The keratin-associated protein 6-1 gene (KAP6-1) is a member of the KAPs family that has a potential correlation with fiber traits. In this study, we investigated the influence of KAP6-1 sequence polymorphisms on the fiber characteristics of a Chinese cashmere-producing goat breed (n = 844). Two main variants were found, including a three base pair (bp) deletion (namely B) and a 36-bp deletion (namely C), while the reference genotype of KAP6-1 was named A. Among them, the B variant was first reported on cashmere goats. This study then correlated these genotypes with the collected fiber data to investigate the potential association of these variants. The results showed that variant A is associated with decreased fiber diameter (p < 0.01), while variant C is associated with deceased fiber length (p < 0.01). These two related variants of the KAP6-1 gene have potential applications as gene-makers to improve the fiber diameter and length in cashmere-producing goats.

High Pulse Pressure Decreases Cardiopulmonary Ability among Older Chinese Patients with Primary Hypertension.

There is limited information regarding the reference values for cardiopulmonary exercise testing (CPX) variables in patients with primary hypertension. Our aim was to provide such values. In this single-center, retrospective study, we included 635 patients (male, 53.7%; mean ± standard deviation age, 62.1 ± 12.6 years) who completed maximal ramp incremental CPX using cycle ergometry at the Rui An Hospital of Traditional Chinese Medicine from August 1, 2018, to December 31, 2021. The patients were classified into two groups based on pulse pressure (PP): (1) patients with a low PP ≤ 60 mmHg (n = 324) and (2) patients with a high PP > 60 mmHg (n = 311). Stepwise linear regression was used to fit the equations of the key CPX variables. CPX was self-interrupted owing to fatigue by 64.5% (low PP) and 72.0% (high PP) of patients (χ2 analysis, P = 0.05). The anaerobic threshold (AT) could not be determined in 3.7% and 7.7%, respectively (χ2 analysis, P = 0.038). Peak oxygen consumption (VO2), carbon dioxide production, ventilation/minute, work rate, and VO2 at the AT were associated with age, weight, and sex. Moreover, a high PP was associated with impaired anaerobic exercise ability, as indicated by the reduced peak VO2, peak VE, and absolute peak work rate. Compared with prior research of a healthy population, patients with primary hypertension in this study exhibited impaired cardiopulmonary ability both at rest and during exercise, especially in the high PP group. The reference values and predictive equations for CPX variables provide a framework for interpreting the response to maximal ramp incremental cycle ergometry among older Chinese patients with primary hypertension.

Tocilizumab for Treatment of Mechanically Ventilated Patients With COVID-19.

BACKGROUND: Severe coronavirus disease 2019 (COVID-19) can manifest in rapid decompensation and respiratory failure with elevated inflammatory markers, consistent with cytokine release syndrome for which IL-6 blockade is an approved treatment. METHODS: We assessed effectiveness and safety of IL-6 blockade with tocilizumab in a single-center cohort of patients with COVID-19 requiring mechanical ventilation. The primary endpoint was survival probability postintubation; secondary analyses included an ordinal illness severity scale integrating superinfections. Outcomes in patients who received tocilizumab compared with tocilizumab-untreated controls were evaluated using multivariable Cox regression with propensity score inverse probability of treatment weighting (IPTW). RESULTS: 154 patients were included, of whom 78 received tocilizumab and 76 did not. Median follow-up was 47 days (range, 28-67). Baseline characteristics were similar between groups, although tocilizumab-treated patients were younger (mean: 55 vs 60 years), less likely to have chronic pulmonary disease (10% vs 28%), and had lower D-dimer values at time of intubation (median: 2.4 vs 6.5 mg/dL). In IPTW-adjusted models, tocilizumab was associated with a 45% reduction in hazard of death (HR, .55; 95% CI, .33-.90) and improved status on the ordinal outcome scale [OR per 1-level increase, .58; .36-.94). Although tocilizumab was associated with an increased proportion of patients with superinfections (54% vs 26%; P < .001), there was no difference in 28-day case fatality rate among tocilizumab-treated patients with versus without superinfection (22% vs 15%; P = .42). Staphylococcus aureus accounted for ~50% of bacterial pneumonia. CONCLUSIONS: In this cohort of mechanically ventilated COVID-19 patients, tocilizumab was associated with lower mortality despite higher superinfection occurrence.

Cadmium exposure enhances organic cation transporter 2 trafficking to the kidney membrane and exacerbates cisplatin nephrotoxicity.

Renal accumulation and exposure of cadmium originating from pollution in agricultural land and the prevalence of cigarette smoking remains an unneglectable human health concern. Whereas cadmium exposure has been correlated with increased incidence of a variety of kidney diseases, little is known pertaining to its effect on renal drug disposition and response in patients. Here, we report that cadmium exposure significantly increased the activity of organic cation transporter 2 (OCT2), a critical renal drug transporter recommended in United States Federal Drug Administration guidance for assessment during drug development. Cadmium enhanced OCT2 trafficking to the cell membrane both in vitro and in vivo. Mechanistically cadmium-mediated OCT2 translocation was found to involve protein-protein interaction between serine/threonine-protein kinase AKT2, calcium/calmodulin and the AKT substrate AS160 in in vitro cellular studies. The formed protein complex could selectively facilitate phosphorylation of AKT2 at T309, which induced translocation of OCT2 to the plasma membrane. Moreover, cadmium exposure markedly exacerbated nephrotoxicity induced by cisplatin, an OCT2 substrate, by increasing its accumulation in the mouse kidney. Consistently, there was a significant correlation between plasma cadmium level and alteration of renal function in cervical cancer patients who underwent chemotherapy with cisplatin. Thus, our studies suggest that membrane transporter distribution induced by cadmium exposure is a previously unrecognized factor for the broad variation in renal drug disposition and response.

Fast and High-Throughput Evaluation of Photodynamic Effect by Monitoring Specific Protein Oxidation with MALDI-TOF Mass Spectrometry.

In antibacterial practices by photodynamic treatment, bacteria are incubated with photosensitizers and then oxidized to death by generating reactive oxygen species (ROS) under light irradiation. Generally, Luria-Bertani (LB) agar colony is a conventional method to evaluate the photodynamic effect. However, this method is time consuming, easily disturbed by pollutants, and limited to the analysis of a pure bacteria sample. Herein, we introduce a novel method of photodynamic effect evaluation through in situ detection of specific protein oxidation by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) with only 1 µL of sample in a fast (less than 1 min per sample) and high-throughput (up to 384 samples per run) way. The oxidation rates of specific proteins stayed highly consistent with bactericidal rates and thus MALDI-TOF MS might be able to replace the LB agar colony to evaluate the photodynamic effect. With the present method, several experimental conditions including different photosensitizer types, dosage controls, and different illumination times were easily screened to optimize photodynamic effect. Photodynamic effects of various bacteria species, cancer cells, and even mixture samples were also evaluated. The results demonstrate the promising application of MALDI-TOF MS in evaluating the photodynamic effect of each component in a mixture sample without any separation or purification, which could not be achieved by the traditional LB agar colony method.

Specific Near-Infrared Probe for Ultrafast Imaging of Lysosomal ß-Galactosidase in Ovarian Cancer Cells.

Reactivity based fluorescent probes have been widely investigated as a powerful and noninvasive tool for disease diagnosis in recent years. ß-Galactosidase (ß-gal), one of the typical lysosomal glycosidases, is reported to be a vital biomarker overexpressed in primary ovarian cancer cells. Fluorescent probes with excellent performance for endogenous ß-gal detection offer a unique option for visualization and diagnosis of primary ovarian cancer cells. Herein, a near-infrared fluorescent probe Lyso-Gal with lysosome-targeting ability was developed for lysosomal ß-gal detection and imaging in ovarian cancer cells (SKOV-3 cells). Lyso-Gal exhibits weak fluorescence in aqueous solution but emits bright NIR fluorescence at 725 nm after incubation with ß-gal. Highly selective imaging of ovarian cancer cells has been achieved upon incubation with Lyso-Gal for only 1 min. The detection time is extremely short. In comparison with a similar hemicyanine probe, Hx-Gal, without lysosome-targeting ability, Lyso-Gal realizes endogenous ß-gal visualization in lysosomes and shows brighter fluorescence than Hx-Gal in SKOV-3 cells. This work demonstrates the potential of Lyso-Gal for detection of primary ovarian cancer cells by using ß-gal as the biomarker.

Highly efficient generation of sheep with a defined FecBB mutation via adenine base editing.

Base editing has the potential to improve important economic traits in agriculture and can precisely convert single nucleotides in DNA or RNA sequences into minimal double-strand DNA breaks (DSB). Adenine base editors (ABE) have recently emerged as a base editing tool for the conversion of targeted A:T to G:C, but have not yet been used in sheep. ABEmax is one of the latest versions of ABE, which consists of a catalytically-impaired nuclease and a laboratory-evolved DNA-adenosine deaminase. The Booroola fecundity (FecBB) mutation (g.A746G, p.Q249R) in the bone morphogenetic protein receptor 1B (BMPR1B) gene influences fecundity in many sheep breeds. In this study, by using ABEmax we successfully obtained lambs with defined point mutations that result in an amino acid substitution (p.Gln249Arg). The efficiency of the defined point mutations was 75% in newborn lambs, since six lambs were heterozygous at the FecBB mutation site (g.A746G, p.Q249R), and two lambs were wild-type. We did not detect off-target mutations in the eight edited lambs. Here, we report the validation of the first gene-edited sheep generated by ABE and highlight its potential to improve economically important traits in livestock.

Performance of the (1,3)-Beta-D-Glucan Assay on Bronchoalveolar Lavage Fluid for the Diagnosis of Invasive Pulmonary Aspergillosis.

Detection of (1,3)-beta-D-glucan (BDG), a component of the cell wall of many fungi, was studied in bronchoalveolar lavage fluid (BALF) as a possible aid for the diagnosis of proven/probable invasive pulmonary aspergillosis (IPA). BDG was measured on stored BALF from 13 patients with EORTC/MSGERC defined proven/probable IPA and 26 matched control patients without IPA. The median BALF BDG was 80 pg/mL (range < 45-8240 pg/mL) in the IPA cohort and 148 pg/mL (range < 45-5460 pg/mL) in the non-IPA cohort. Using a positive cutoff of ≥ 80 pg/mL, sensitivity was 54% and specificity was 38%. Higher cutoff values led to improvement in specificity but a dramatic decrease in sensitivity. ROC/AUC analysis was unable to identify an optimal cutoff value at which test performance was enhanced: AUC 0.43, 95% CI 0.24-0.63. When the BDG assay was performed on BALF, neither sensitivity nor specificity was sufficient for use in the diagnosis of IPA.

Breakthrough Invasive Fungal Infections in Patients with Acute Myeloid Leukemia.

OBJECTIVES: We sought to determine the occurrence, risk factors, effect of antifungal prophylaxis, and outcomes of invasive fungal infections (IFIs) in patients with acute myeloid leukemia (AML). PATIENTS AND METHODS: We performed a retrospective analysis of all adult patients admitted to the University of Michigan Health System for AML over a 3-year period from 2010 to 2013. We determined comorbidities, hematopoietic cell transplant (HCT) status, antifungal prophylaxis, proven and probable IFI, and outcomes at 12 weeks after initiation of appropriate antifungal therapy. RESULTS: Of 333 patients in our cohort, 116 of whom had received a HCT, 98 (29%) developed an IFI. Of the 30 (9%) patients who had a proven or probable IFI, 18 had breakthrough infection while on micafungin (n = 5), voriconazole (n = 4), posaconazole (n = 5), or fluconazole (n = 4). Breakthrough IFIs were due to Aspergillus species (n = 11), other molds (n = 4), and Candida species (n = 3). Factors associated with breakthrough IFI were prolonged severe neutropenia (p = .05) and having received tacrolimus (p = .04). Antifungal therapy was successful in 7 of the 18 (39%) patients with breakthrough IFI and 8 of the 12 (67%) patients with non-breakthrough IFI, p = .13. Mortality at 12 weeks was 27%, 5 with breakthrough IFI and 3 with non-breakthrough IFI and was associated with prolonged severe neutropenia, p = .04. CONCLUSIONS: Patients with AML remain at risk for IFI despite the use of several different antifungal agents for prophylaxis. Mortality remains high in patients with AML who develop IFI.

Multidrug resistant organism predicts ulcer recurrence following surgical management of diabetic foot osteomyelitis.

Diabetic foot ulcers commonly precede diabetic foot osteomyelitis (DFO) and once the latter occurs, surgical management is often performed. The presence of osteomyelitis is an independent risk factor for the development of re-ulceration. We investigated the relationship between causative organisms in osteomyelitis and 1-year diabetic foot outcomes (re-ulceration, amputation, and death) following surgical management in an observational cohort of 223 patients. Univariate and multivariate analyses were performed for available demographic, clinical, and laboratory data. In addition, random forest plots were used to identify microbiologic predictors of 1-year outcomes. Patients with osteomyelitis managed surgically were younger and exhibited more painful peripheral neuropathy than outpatients with diabetes alone (both P < .0001). Osteomyelitis proximal margin cultures were diverse, at times polymicrobial, and included multidrug-resistant organisms in 13.9% of the cohort. In patients who underwent surgery, 44.5% experienced a re-ulceration on the same foot within 12 months of surgery. The presence of multidrug-resistant organisms on proximal bone culture was found to be a significant predictor of diabetic foot ulcer recurrence in univariate modelling (P < .001) and importance rankings. This is the first study to use prediction modelling to identify a relationship between multidrug-resistant organisms and diabetic foot ulcer recurrence following DFO.