Multiparatopic antibodies induce targeted downregulation of programmed death-ligand 1.

Programmed death-ligand 1 (PD-L1) drives inhibition of antigen-specific T cell responses through engagement of its receptor programmed death-1 (PD-1) on activated T cells. Overexpression of these immune checkpoint proteins in the tumor microenvironment has motivated the design of targeted antibodies that disrupt this interaction. Despite clinical success of these antibodies, response rates remain low, necessitating novel approaches to enhance performance. Here, we report the development of antibody fusion proteins that block immune checkpoint pathways through a distinct mechanism targeting molecular trafficking. By engaging multiple receptor epitopes on PD-L1, our engineered multiparatopic antibodies induce rapid clustering, internalization, and degradation in an epitope- and topology-dependent manner. The complementary mechanisms of ligand blockade and receptor downregulation led to more durable immune cell activation and dramatically reduced PD-L1 availability in mouse tumors. Collectively, these multiparatopic antibodies offer mechanistic insight into immune checkpoint protein trafficking and how it may be manipulated to reprogram immune outcomes.

Heat stress from current and predicted increases in temperature impairs lambing rates and birth weights in the Australian sheep flock.

Livestock heat stress threatens production, particularly in semi-arid, arid and tropical regions. Using established temperature thresholds for sheep, we modelled +1 °C and +3 °C temperature increases over the historical baseline, estimating that 2.1 million potential lambs are lost annually due to heat stress alone, increasing to 2.5 and 3.3 million, respectively, as temperatures rise. Heat stress poses risks at key periods of the reproductive cycle, with consequences across the Australian sheep flock.

Tracking Cancer: Exploring Heart Rate Variability Patterns by Cancer Location and Progression.

Reduced heart rate variability (HRV) is an autonomic nervous system (ANS) response that may indicate dysfunction in the human body. Consistent evidence shows cancer patients elicit lower HRV; however, only select cancer locations were previously evaluated. Thus, the aim of the current study was to explore HRV patterns in patients diagnosed with and in varying stages of the most prevalent cancers. At a single tertiary academic medical center, 798 patients were recruited. HRV was measured via an armband monitor (Warfighter MonitorTM, Tiger Tech Solutions, Inc., Miami, FL, USA) equipped with electrocardiographic capabilities and was recorded for 5 to 7 min with patients seated in an upright position. Three time-domain metrics were calculated: SDNN (standard deviation of the NN interval), rMSSD (the root mean square of successive differences of NN intervals), and the percentage of time in which the change in successive NN intervals exceeds 50ms within a measurement (pNN50). Of the 798 patients, 399 were diagnosed with cancer. Cancer diagnoses were obtained via medical records one week following the measurement. Analysis of variance models were performed comparing the HRV patterns between different cancers, cancer stages (I-IV), and demographic strata. A total of 85% of the cancer patients had breast, gastrointestinal, genitourinary, or respiratory cancer. The cancer patients were compared to a control non-cancer patient population with similar patient size and distributions for sex, age, body mass index, and co-morbidities. For all HRV metrics, non-cancer patients exhibited significantly higher rMSSDs (11.1 to 13.9 ms, p < 0.0001), SDNNs (22.8 to 27.7 ms, p < 0.0001), and pNN50s (6.2 to 8.1%, p < 0.0001) compared to stage I or II cancer patients. This significant trend was consistently observed across each cancer location. Similarly, compared to patients with stage III or IV cancer, non-cancer patients possessed lower HRs (-11.8 to -14.0 bpm, p < 0.0001) and higher rMSSDs (+31.7 to +32.8 ms, p < 0.0001), SDNNs (+45.2 to +45.8 ms), p < 0.0001, and pNN50s (19.2 to 21.6%, p < 0.0001). The HR and HRV patterns observed did not significantly differ between cancer locations (p = 0.96 to 1.00). The depressed HRVs observed uniformly across the most prevalent cancer locations and stages appeared to occur independent of patients' co-morbidities. This finding highlights the potentially effective use of HRV as a non-invasive tool for determining common cancer locations and their respective stages. More studies are needed to delineate the HRV patterns across different ages, between sexes and race/ethnic groups.

Deep learning enables satellite-based monitoring of large populations of terrestrial mammals across heterogeneous landscape.

New satellite remote sensing and machine learning techniques offer untapped possibilities to monitor global biodiversity with unprecedented speed and precision. These efficiencies promise to reveal novel ecological insights at spatial scales which are germane to the management of populations and entire ecosystems. Here, we present a robust transferable deep learning pipeline to automatically locate and count large herds of migratory ungulates (wildebeest and zebra) in the Serengeti-Mara ecosystem using fine-resolution (38-50 cm) satellite imagery. The results achieve accurate detection of nearly 500,000 individuals across thousands of square kilometers and multiple habitat types, with an overall F1-score of 84.75% (Precision: 87.85%, Recall: 81.86%). This research demonstrates the capability of satellite remote sensing and machine learning techniques to automatically and accurately count very large populations of terrestrial mammals across a highly heterogeneous landscape. We also discuss the potential for satellite-derived species detections to advance basic understanding of animal behavior and ecology.

COVID-19-Induced Changes in Photoplethysmography.

INTRODUCTION: Photoplethysmography (PPG) is the science behind many commonly used medical devices such as the pulse oximeter. PPG changes, herein as "PPG dropouts," have been described in existing in vitro studies following artificially induced clot activation. Because COVID-19 causes increased arterial, venous, and microvascular clot formation, our hypothesis is that PPG dropouts identified in vitro can also be found in vivo in patients with COVID-19. The aim of this study is to evaluate PPG recordings and D-dimer levels for patients hospitalized with COVID-19 and compare them with the PPG tracings from non-COVID controls. MATERIALS AND METHODS: PPG recordings were obtained for 197 ICU patients with COVID-19 and 300 non-COVID controls. PPG tracings were obtained using a TigerTech CovidPlus monitor, which received U.S. FDA emergency use authorization in March 2020 for monitoring the biometrics of patients with COVID-19 and featured unfiltered red and infrared spectrum PPG monitoring. D-dimer lab results were also recorded whenever available. RESULTS: The results demonstrated significant differences in the prevalence rate of PPG dropout among patients with COVID-19 vs. non-COVID controls. The median PPG dropout rate was 0.58 for COVID-19 patients (median 0.58, IQR 0.42-0.72, P < .05) as opposed to a median 0.0 for non-COVID patients (median 0.0, IQR 0.0-0.0, P < .05). Furthermore, at least one incidence of PPG dropout was detected in 100% of COVID-19 patients, as opposed to 2.3% of non-COVID controls (P < .05). PPG dropout also correlated closely with the normalized serum D-dimer levels taken on the same day. The change in the normalized D-dimer levels was plotted against the change in PPG dropout, and a line of best fit was created. Linear regression resulted in R2 = 0.743 (P < .05), indicating that changes in the PPG dropout rate correlate with hemorheological changes in COVID-19 patients. CONCLUSIONS: PPG dropout, like D-dimer, may not be specific for COVID-19. However, the inflammatory nature of the disease and the prevalence of prolonged ICU created a large sample size and allowed the authors to observe PPG changes in vivo in a statistically meaningful way. Further confirmatory studies are needed to confirm the potential application of PPG dropout as a measure of inflammation in other disease processes.

Internal or External Training Load Metrics: Which Is Best for Tracking Autonomic Nervous System Recovery and Function in Collegiate American Football?

Sport coaches increasingly rely on external load metrics for designing effective training programs. However, their accuracy in estimating internal load is inconsistent, and their ability to predict autonomic nervous system (ANS) deterioration is unknown. This study aimed to evaluate the relationships between internal and external training load metrics and ANS recovery and function in college football players. Football athletes were recruited from a D1 college in the southeastern US and prospectively followed for 27 weeks. Internal load was estimated via exercise cardiac load (ECL; average training heartrate (HR) × session duration) and measured with an armband monitor equipped with electrocardiographic capabilities (Warfighter MonitorTM (WFM), Tiger Tech Solutions, Miami, FL, USA). External load was estimated via the summation and rate of acceleration and decelerations as measured by a triaxial accelerometer using the WFM and an accelerometer-based (ACCEL) device (Catapult Player Load, Catapult Sports, Melbourne, Australia) worn on the mid-upper back. Baseline HR, HR variability (HRV) and HR recovery served as the indicators for ANS recovery and function, respectively. For HRV, two, time-domain metrics were measured: the standard deviation of the NN interval (SDNN) and root mean square of the standard deviation of the NN interval (rMSSD). Linear regression models evaluated the associations between ECL, ACCEL, and the indicators of ANS recovery and function acutely (24 h) and cumulatively (one- and two-week). Athletes (n = 71) were male and, on average, 21.3 ± 1.4 years of age. Acute ECL elicited stronger associations for 24 h baseline HR (R2 0.19 vs. 0.03), HR recovery (R2 0.38 vs. 0.07), SDNN (R2 0.19 vs. 0.02) and rMSSD (R2 0.19 vs. 0.02) compared to ACCEL. Similar results were found for one-week: 24 h baseline HR (R2 0.48 vs. 0.05), HR recovery (R2 0.55 vs. 0.05), SDNN (R2 0.47 vs. 0.05) and rMSSD (R2 0.47 vs. 0.05) and two-week cumulative exposures: 24 h baseline HR (R2 0.52 vs. 0.003), HR recovery (R2 0.57 vs. 0.05), SDNN (R2 0.52 vs. 0.003) and rMSSD (R2 0.52 vs. 0.002). Lastly, the ACCEL devices weakly correlated with ECL (rho = 0.47 and 0.43, p < 0.005). Our findings demonstrate that ACCEL poorly predicted ANS deterioration and underestimated internal training load. ACCEL devices may "miss" the finite window for preventing ANS deterioration by potentially misestimating training loads acutely and cumulatively.

Poly(Beta-Amino Ester)s as High-Yield Transfection Reagents for Recombinant Protein Production.

Purpose: Transient transfection is an essential tool for recombinant protein production, as it allows rapid screening for expression without stable integration of genetic material into a target cell genome. Poly(ethylenimine) (PEI) is the current gold standard for transient gene transfer, but transfection efficiency and the resulting protein yield are limited by the polymer's toxicity. This study investigated the use of a class of cationic polymers, poly(beta-amino ester)s (PBAEs), as reagents for transient transfection in comparison to linear 25 kDa PEI, a commonly used transfection reagent. Methods: Transfection efficiency and protein production were assessed in human embryonic kidney 293F (HEK) and Chinese hamster ovary-S (CHO) cell suspensions using PBAE-based nanoparticles in comparison to linear 25 kDa PEI. Production of both a cytosolic reporter and secreted antibodies was investigated. Results: In both HEK and CHO cells, several PBAEs demonstrated superior transfection efficiency and enhanced production of a cytosolic reporter compared to linear 25 kDa PEI. This result extended to secreted proteins, as a model PBAE increased the production of 3 different secreted antibodies compared to linear 25 kDa PEI at culture scales ranging from 20 to 2000 mL. In particular, non-viral gene transfer using the lead PBAE/plasmid DNA nanoparticle formulation led to robust transfection of mammalian cells across different constructs, doses, volumes, and cell types. Conclusion: These results show that PBAEs enhance transfection efficiency and increase protein yield compared to a widespread commercially available reagent, making them attractive candidates as reagents for use in recombinant protein production.

Borane- and Silylium-Catalyzed Difunctionalization of Carbohydrates: 3,6-Anhydrosugar Enabled 1,6-Site Selectivity.

A novel diastereoselective, Lewis acid catalyzed 1,6-difunctionalization of galactose and mannose derivatives has been developed in one pot, via sequential nucleophile additions. Our studies point to the formation of a 3,6-anhydrosugar intermediate as key to the 1,6-site-selectivity. Starting material-specific reactivity occurs when competitive ring-opening C-O cleavage is possible, owed to basicity and stereoelectronic stabilization differences. Lastly, Mayr nucleophilicity parameter values helped predict which reaction conditions would be most suitable for specific nucleophiles.

No COVID-19 climate silver lining in the US power sector.

Recent studies conclude that the global coronavirus (COVID-19) pandemic decreased power sector CO2 emissions globally and in the United States. In this paper, we analyze the statistical significance of CO2 emissions reductions in the U.S. power sector from March through December 2020. We use Gaussian process (GP) regression to assess whether CO2 emissions reductions would have occurred with reasonable probability in the absence of COVID-19 considering uncertainty due to factors unrelated to the pandemic and adjusting for weather, seasonality, and recent emissions trends. We find that monthly CO2 emissions reductions are only statistically significant in April and May 2020 considering hypothesis tests at 5% significance levels. Separately, we consider the potential impact of COVID-19 on coal-fired power plant retirements through 2022. We find that only a small percentage of U.S. coal power plants are at risk of retirement due to a possible COVID-19-related sustained reduction in electricity demand and prices. We observe and anticipate a return to pre-COVID-19 CO2 emissions in the U.S. power sector.

Single limb electrocardiogram using vector mapping: Evaluation and validation of a novel medical device.

INTRODUCTION: The Tiger Tech Warfighter Monitor (WFM) is a novel single-limb device for ECG acquisition. The WFM provides true (not derived) single limb Electrocardiogram monitoring (ECG) to provide heart rate and R-R interval monitoring between QRS complexes. Herein, we evaluate the diagnostic accuracy of the WFM heart rate, R-R interval monitoring, and heart rate variability monitoring in comparison to a 2­lead chest ECG. METHODS: Data was collected under Institutional Review Board (IRB) approval. Patients available within our institution's pre-operative holding unit were randomly selected to undergo simultaneous chest and WFM ECG monitoring. 3-5-min measurements were taken depending on the patient's availability. Data was saved to two separate mobile phones and time-stamped for synchronization. A proprietary Tiger Tech extraction algorithm was used to tag proper features on both the WFM 1-Limb ECG and Chest ECG data files. A separate algorithm was then used to compare the beat-to-beat variations between the ECGs. RESULTS: Data was extracted and analyzed on 26 subjects. Linear regression of heart rate analysis revealed excellent correlations with an R2 of 0.99 (p < 0.05). Similar linear regression evaluation of R-R interval correlation demonstrated a mean R2 value of 0.95 (p < 0.05). Statistically significant correlation was achieved in all 26 included study participants. Heart rate variability also achieved excellent correlation (SDNN R2 = 0.997, RMSSD R2 = 0.995, LnRMSSD R2 = 0.992, p << 0.05). CONCLUSION: Results demonstrate that the WFM achieves excellent correlation with chest ECG for heart rate, R-R internals, and heart rate variability.

Evolution of core archetypal phenotypes in progressive high grade serous ovarian cancer.

The evolution of resistance in high-grade serous ovarian cancer (HGSOC) cells following chemotherapy is only partially understood. To understand the selection of factors driving heterogeneity before and through adaptation to treatment, we profile single-cell RNA-sequencing (scRNA-seq) transcriptomes of HGSOC tumors collected longitudinally during therapy. We analyze scRNA-seq data from two independent patient cohorts to reveal that HGSOC is driven by three archetypal phenotypes, defined as oncogenic states that describe the majority of the transcriptome variation. Using a multi-task learning approach to identify the biological tasks of each archetype, we identify metabolism and proliferation, cellular defense response, and DNA repair signaling as consistent cell states found across patients. Our analysis demonstrates a shift in favor of the metabolism and proliferation archetype versus cellular defense response archetype in cancer cells that received multiple lines of treatment. While archetypes are not consistently associated with specific whole-genome driver mutations, they are closely associated with subclonal populations at the single-cell level, indicating that subclones within a tumor often specialize in unique biological tasks. Our study reveals the core archetypes found in progressive HGSOC and shows consistent enrichment of subclones with the metabolism and proliferation archetype as resistance is acquired to multiple lines of therapy.

Synthetic Biology towards Engineering Microbial Lignin Biotransformation.

Lignin is the second most abundant biopolymer on earth and is a major source of aromatic compounds; however, it is vastly underutilized owing to its heterogeneous and recalcitrant nature. Microorganisms have evolved efficient mechanisms that overcome these challenges to depolymerize lignin and funnel complex mixtures of lignin-derived monomers to central metabolites. This review summarizes recent synthetic biology efforts to enhance lignin depolymerization and aromatic catabolism in bacterial and fungal hosts for the production of both natural and novel bioproducts. We also highlight difficulties in engineering complex phenotypes and discuss the outlook for the future of lignin biological valorization.

Reduced renewable energy stability in India following COVID-19: Insights and key policy recommendations.

The COVID-19 pandemic has dramatically altered global energy consumption, particularly affecting investment in renewable energy projects. In India, strict shelter-in-place orders enforced during March 2020 have since led to a considerable change in public and private sector investments in planned renewable energy installations. In this paper, we attempt to highlight trends in energy consumption and installed renewable energy capacity noted in India during a period concurrent with the shelter-in-place orders. We discuss recent policy measures and additions to installed renewable energy capacity, and propose key policy recommendations that may help the sector adopt a growth trajectory similar to one noted pre-pandemic. This paper is organized into four main parts. In the first section, we draw focus to India's renewable energy policies and pay special emphasis on recent interventions and campaigns targeted towards achieving high growth rates in the sector. We briefly discuss the need for effective public-private partnerships in order to meet these targets. In the second part, we quantitatively characterise the growth of renewables in India. We present an overview of several mechanisms and missions the government has launched in line with their policy to mitigate the environmental impact of India's energy mix. In the third part, we analyse the decrease in electricity demand in India from 24 March to 30 June 2020, a period concurrent with shelter-at-home orders issued by the Government. We also characterise changes in installed renewable energy capacity between March to December 2016-2020 to provide causal evidence of the effect of the pandemic on the growth of renewables. In this section, we also compile and analyse data on state-wise stressed assets across renewable energy generators in the country. Lastly, in the fourth and final portion of this paper, we highlight policy recommendations that may help the sector overcome logistical and financial bottlenecks in the short-term. We do this with the hope of outlining key measures that decision makers may employ to achieve pre-COVID sectoral growth in the long term. Our recommendations cover three different policy instruments: investment subsidies, operational subsidies, and recommendations for DISCOMs.

Prolapse repair after anterior exenteration.

This report presents our experience in performing prolapse repair after anterior exenteration. The patient had a history of invasive bladder cancer and underwent a robotically assisted laparoscopic anterior exenteration with extended bilateral pelvic lymph node dissection and creation of an Indiana pouch continent diversion. Her pelvic organ prolapse progressed over time despite multiple pessary fittings. She eventually decided to proceed with pelvic reconstructive surgery 6 years after her cancer surgery. She underwent a successful vaginal native tissue reconstruction with uterosacral ligament suspension, posterior repair and reconstruction of the anterior compartment. The patient has been followed for 16 months without recurrent prolapse. Vaginal native tissue pelvic reconstruction is feasible in a patient with a history of pelvic exenteration.

Silylpalladium Cations Enable the Oxidative Addition of C(sp3)-O Bonds.

The synthesis and characterization of the room-temperature and solution-stable silylpalladium cations (PCy3)2Pd-SiR3+(C6F5)4B- (SiR3 = SiMe2Et, SiHEt2) and (Xantphos)Pd-SiR3+(BArf4) (SiR3 = SiMe2Et, SiHEt2; Xantphos = 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene; BArf4 = (3,5-(CF3)2C6H3)4B-) are reported. Spectroscopic and ligand addition experiments suggest that silylpalladium complexes of the type (PCy3)2Pd-SiR3+ are three-coordinate and T-shaped. Addition of dialkyl ethers to both the PCy3 and Xantphos-based silylpalladium cations resulted in the cleavage of C(sp3)-O bonds and the generation of cationic Pd-alkyl complexes. Mechanistically enabling is the ability of silylpalladium cations to behave as sources of both electrophilic silylium ions and nucleophilic LnPd(0).

Site Selective Amide Reduction of Cyclosporine A Enables Diverse Derivation of an Important Cyclic Peptide.

Site selective amide reductions of the cyclic undecapeptide, cyclosporine A, have been developed using the combination of a heteroleptic borane catalyst and a silane reductant. Tertiary silane Me2EtSiH provides two unique cyclosporine A derivatives, one of which can be readily diversified in subsequent reactions. The secondary silane Et2SiH2 enables divergent reactivity that uses a free hydroxyl group to direct the reduction. The transient O-silyl hemiaminal intermediate of this reduction can additionally be trapped by reducing to the amine or by reductive cyanation.

Concurrent pelvic reconstruction and minimally invasive pelvic cancer surgery.

INTRODUCTION AND HYPOTHESIS: We present our experience in performing concurrent prolpase repair at the time of gynecologic cancer surgery. METHODS: The uterosacral ligaments are tagged before performing hysterectomy and pelvic dissection. The uterosacral ligament suspensory sutures are then placed laparoscopically after completion of pelvic cancer surgery. The remainder of the prolapse surgery is performed through a transvaginal approach. RESULTS: Many of our patients who undergo concurrent prolapse repair and gynecolgical cancer surgery receive chemotherapy and pelivc radiation. Concuurent prolapse repair improves their prolaspe symptoms. CONCLUSION: Concurrent prolapse repair should be performed at the same time as gynecologic cancer surgery.

A collaborative surgical approach to upper and lower abdominal cytoreductive surgery in ovarian cancer.

BACKGROUND AND OBJECTIVES: Cytoreductive surgery with complete macroscopic resection in patients with ovarian cancer is associated with improved survival. Institutional reports of combined upper and lower abdominal cytoreductive surgery for more advanced disease have described multidisciplinary approaches. We sought to investigate outcomes in patients undergoing cytoreductive surgery in patients with upper and lower abdominal disease at our institution. METHODS: Patients who underwent cytoreductive surgery for ovarian malignancies from 2008 to 2015 were retrospectively identified from an institutional database. Upper abdominal cytoreduction was defined anatomically as debulking of disease proximal to the ligament of Treitz. Perioperative outcomes were analyzed. RESULTS: A total of 258 operations were performed, the majority for serous ovarian carcinoma (70%). The gynecologic oncologist was the primary surgeon and often assisted by either a surgical oncology fellow and/or attending. In operations with combined upper and lower abdominal cytoreduction, patients were more likely to have an American society of anesthesiologists physical status classification system (ASA) of 3, peritoneal implants, and liver/spleen metastases. Preoperative chemotherapy and optimal cytoreduction were similar between groups. Perioperative morbidity and mortality were not significantly different between groups. CONCLUSIONS: A collaborative surgical approach to combined upper and lower abdominal cytoreductive surgery in patients with ovarian cancer should be performed, if needed, to achieve an optimal cytoreduction.