Therapeutic Myths in Solid Organ Transplantation Infectious Diseases.

Infection management in solid organ transplantation poses unique challenges, with a diverse array of potential pathogens and associated antimicrobial therapies. With limited high-quality randomized clinical trials to direct optimal care, therapeutic "myths" may propagate and contribute to suboptimal or excessive antimicrobial use. We discuss 6 therapeutic myths with particular relevance to solid organ transplantation and provide recommendations for infectious diseases clinicians involved in the care of this high-risk population.

Scuba diving-related fatalities in the Philippines from 2008 to 2022 as reported in online news media.

The present study aims to investigate the demographics and characteristics of scuba diving fatalities in the Philippines which can help in the identification of local trends and ultimately in the development of appropriate preventive measures. Data on scuba diving-related fatalities in the Philippines from 2008 to 2022 were manually retrieved from online news media sources. Information on age, sex, nationality, certification, purpose, and causative factors, whenever possible were collected and analysed. A total of 39 fatalities were identified having a median age of 43.5 (range 20-80). Majority of victims were males (n = 30), and of foreign ethnicity (n = 26). Asphyxia was identified as the possible disabling injury in almost half of the cases (n = 17). The causes of death based on autopsies were determined only for few cases which included drowning (n = 2), heart attack (n = 1), and traumatic injuries from a dynamite blast (n = 1). Potential vulnerable groups were identified to be the ageing population and foreign tourist divers. In the absence of an existing database, this preliminary report provides the best available evidence at this time concerning scuba diving fatalities in the Philippines.

Permittivity tensor imaging: modular label-free imaging of 3D dry mass and 3D orientation at high resolution.

The dry mass and the orientation of biomolecules can be imaged without a label by measuring their permittivity tensor (PT), which describes how biomolecules affect the phase and polarization of light. Three-dimensional (3D) imaging of PT has been challenging. We present a label-free computational microscopy technique, PT imaging (PTI), for the 3D measurement of PT. PTI encodes the invisible PT into images using oblique illumination, polarization-sensitive detection and volumetric sampling. PT is decoded from the data with a vectorial imaging model and a multi-channel inverse algorithm, assuming uniaxial symmetry in each voxel. We demonstrate high-resolution imaging of PT of isotropic beads, anisotropic glass targets, mouse brain tissue, infected cells and histology slides. PTI outperforms previous label-free imaging techniques such as vector tomography, ptychography and light-field imaging in resolving the 3D orientation and symmetry of organelles, cells and tissue. We provide open-source software and modular hardware to enable the adoption of the method.

MultiSero: An Open-Source Multiplex-ELISA Platform for Measuring Antibody Responses to Infection.

A multiplexed enzyme-linked immunosorbent assay (ELISA) that simultaneously measures antibody binding to multiple antigens can extend the impact of serosurveillance studies, particularly if the assay approaches the simplicity, robustness, and accuracy of a conventional single-antigen ELISA. Here, we report on the development of multiSero, an open-source multiplex ELISA platform for measuring antibody responses to viral infection. Our assay consists of three parts: (1) an ELISA against an array of proteins in a 96-well format; (2) automated imaging of each well of the ELISA array using an open-source plate reader; and (3) automated measurement of optical densities for each protein within the array using an open-source analysis pipeline. We validated the platform by comparing antibody binding to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) antigens in 217 human sera samples, showing high sensitivity (0.978), specificity (0.977), positive predictive value (0.978), and negative predictive value (0.977) for classifying seropositivity, a high correlation of multiSero determined antibody titers with commercially available SARS-CoV-2 antibody tests, and antigen-specific changes in antibody titer dynamics upon vaccination. The open-source format and accessibility of our multiSero platform can contribute to the adoption of multiplexed ELISA arrays for serosurveillance studies, for SARS-CoV-2 and other pathogens of significance.

A systematic review of direct acting antiviral therapies in hepatitis C virus-negative liver transplant recipients of hepatitis C-viremic donors.

The introduction of safe and highly effective direct acting antivirals (DAAs) has significantly improved hepatitis C virus (HCV) treatment outcomes after transplant. The solid organ transplant community has sought to identify strategies aimed at increasing the donor pool including the utilization of HCV-viremic organs in HCV-negative recipients. We will review the existing literature to evaluate DAA use for the treatment of HCV viremia post-liver transplant in patients who receive HCV-viremic allografts. A PubMed search was conducted and references for each study were also reviewed to identify additional articles. Randomized controlled trials, cohort studies, case series, and case reports were included if: published in English language, evaluated DAA treatment outcomes after liver only or simultaneous liver-kidney transplantation with HCV-viremic allografts in HCV-negative recipients, and had full-text article availability. Our review included 16 studies and 2 case reports. The majority of liver transplant recipients were treated with a pangenotypic DAA for 12 weeks with a heterogeneous median time to initiation (range 1.7-118 days). Sustained virologic response was assessed in 253 liver transplant patients with 99.6% achieving cure with minimal DAA-attributed adverse drug events. There were 23 reported episodes of rejection, 12 deaths, and 1 graft loss among all studies. Treatment with DAA after transplantation of HCV-viremic livers into HCV-negative recipients appears to be safe and effective; however, long-term outcomes remain unknown. Transplant pharmacists play a key role in the development of center-specific protocols to optimize post-transplant outcomes in this unique patient population.

An investigation of aquatic oil spills in the Philippines from 2000 to 2021.

As an archipelago, the Philippines is heavily engaged in domestic and foreign shipping activities which makes it highly vulnerable to oil pollution. It is the purpose of this present paper to make a follow-up review of the oil spill situation in the Philippines which provides analysis and evaluation of aquatic oil spills in the past 22 years. Results showed that the frequency and volume of oil spills generally occur in areas with high maritime traffic and experienced short-term decreases on periods affected by economic recessions. The sources and causes of oil spills are discussed while examining the possible influences of the changing climate. While, these had been identified, there is a gap in the incident reporting of oil spills. It is recommended that data records on oil spill incidents in the Philippines be systematic, consistent and comprehensively entail information not limited to date, locality, source, cause, and spillage amount.

OpenCell: Endogenous tagging for the cartography of human cellular organization.

Elucidating the wiring diagram of the human cell is a central goal of the postgenomic era. We combined genome engineering, confocal live-cell imaging, mass spectrometry, and data science to systematically map the localization and interactions of human proteins. Our approach provides a data-driven description of the molecular and spatial networks that organize the proteome. Unsupervised clustering of these networks delineates functional communities that facilitate biological discovery. We found that remarkably precise functional information can be derived from protein localization patterns, which often contain enough information to identify molecular interactions, and that RNA binding proteins form a specific subgroup defined by unique interaction and localization properties. Paired with a fully interactive website (opencell.czbiohub.org), our work constitutes a resource for the quantitative cartography of human cellular organization.

DynaMorph: self-supervised learning of morphodynamic states of live cells.

A cell's shape and motion represent fundamental aspects of cell identity and can be highly predictive of function and pathology. However, automated analysis of the morphodynamic states remains challenging for most cell types, especially primary human cells where genetic labeling may not be feasible. To enable automated and quantitative analysis of morphodynamic states, we developed DynaMorph-a computational framework that combines quantitative live cell imaging with self-supervised learning. To demonstrate the robustness and utility of this approach, we used DynaMorph to annotate morphodynamic states observed with label-free measurements of optical density and anisotropy of live microglia isolated from human brain tissue. These cells show complex behavior and have varied responses to disease-relevant perturbations. DynaMorph generates quantitative morphodynamic representations that can be used to compare the effects of the perturbations. Using DynaMorph, we identify distinct morphodynamic states of microglia polarization and detect rare transition events between states. The concepts and the methods presented here can facilitate automated discovery of functional states of diverse cellular systems.

Human microglia states are conserved across experimental models and regulate neural stem cell responses in chimeric organoids.

Microglia are resident macrophages in the brain that emerge in early development and respond to the local environment by altering their molecular and phenotypic states. Fundamental questions about microglia diversity and function during development remain unanswered because we lack experimental strategies to interrogate their interactions with other cell types and responses to perturbations ex vivo. We compared human microglia states across culture models, including cultured primary and pluripotent stem cell-derived microglia. We developed a "report card" of gene expression signatures across these distinct models to facilitate characterization of their responses across experimental models, perturbations, and disease conditions. Xenotransplantation of human microglia into cerebral organoids allowed us to characterize key transcriptional programs of developing microglia in vitro and reveal that microglia induce transcriptional changes in neural stem cells and decrease interferon signaling response genes. Microglia additionally accelerate the emergence of synchronized oscillatory network activity in brain organoids by modulating synaptic density.

multiSero: open multiplex-ELISA platform for analyzing antibody responses to SARS-CoV-2 infection.

Serology has provided valuable diagnostic and epidemiological data on antibody responses to SARS-CoV-2 in diverse patient cohorts. Deployment of high content, multiplex serology platforms across the world, including in low and medium income countries, can accelerate longitudinal epidemiological surveys. Here we report multiSero, an open platform to enable multiplex serology with up to 48 antigens in a 96-well format. The platform consists of three components: ELISA-array of printed proteins, a commercial or home-built plate reader, and modular python software for automated analysis (pysero). We validate the platform by comparing antibody titers against the SARS-CoV-2 Spike, receptor binding domain (RBD), and nucleocapsid (N) in 114 sera from COVID-19 positive individuals and 87 pre-pandemic COVID-19 negative sera. We report data with both a commercial plate reader and an inexpensive, open plate reader (nautilus). Receiver operating characteristic (ROC) analysis of classification with single antigens shows that Spike and RBD classify positive and negative sera with the highest sensitivity at a given specificity. The platform distinguished positive sera from negative sera when the reactivity of the sera was equivalent to the binding of 1 ng mL âˆ'1 RBD-specific monoclonal antibody. We developed normalization and classification methods to pool antibody responses from multiple antigens and multiple experiments. Our results demonstrate a performant and accessible pipeline for multiplexed ELISA ready for multiple applications, including serosurveillance, identification of viral proteins that elicit antibody responses, differential diagnosis of circulating pathogens, and immune responses to vaccines.

multiSero: open multiplex-ELISA platform for analyzing antibody responses to SARS-CoV-2 infection

Serology has provided valuable diagnostic and epidemiological data on antibody responses to SARS-CoV-2 in diverse patient cohorts. Deployment of high content, multiplex serology platforms across the world, including in low and medium income countries, can accelerate longitudinal epidemiological surveys. Here we report multiSero, an open platform to enable multiplex serology with up to 48 antigens in a 96-well format. The platform consists of three components: ELISA-array of printed proteins, a commercial or home-built plate reader, and modular python software for automated analysis (pysero). We validate the platform by comparing antibody titers against the SARS-CoV-2 Spike, receptor binding domain (RBD), and nucleocapsid (N) in 114 sera from COVID-19 positive individuals and 87 pre-pandemic COVID-19 negative sera. We report data with both a commercial plate reader and an inexpensive, open plate reader (nautilus). Receiver operating characteristic (ROC) analysis of classification with single antigens shows that Spike and RBD classify positive and negative sera with the highest sensitivity at a given specificity. The platform distinguished positive sera from negative sera when the reactivity of the sera was equivalent to the binding of 1 ng mL-1 RBD-specific monoclonal antibody. We developed normalization and classification methods to pool antibody responses from multiple antigens and multiple experiments. Our results demonstrate a performant and accessible pipeline for multiplexed ELISA ready for multiple applications, including serosurveillance, identification of viral proteins that elicit antibody responses, differential diagnosis of circulating pathogens, and immune responses to vaccines.

Superficial temporal artery perforator flaps for reconstruction of intraoral defects.

BACKGROUND: Intraoral defects after tumor resection are often reconstructed with free tissue transfer. However, in patients who are not good candidates for free tissue transfer, regional flaps based on the superficial temporal artery can be utilized. The authors present our technique to reconstruct intraoral defects with the superficial temporal artery perforator (STAP) flap and early outcomes. METHODS: Five patients underwent STAP flaps for defects including the hard palate, buccal sulcus, floor of mouth, and retromolar trigone between 2017 and 2019. The mean defect size was 5.6 × 3.4 cm2 (3 × 3 cm2 - 7 × 4 cm2 ). The mean age was 74 (57-88) and all patients had recurrent cancer. External Doppler, indocyanine green laser angiography, and FLIR thermal imaging were used intra-operatively to identify the best perforators and plan for flap design. RESULTS: The mean flap size was 7.6 × 3.5 cm2 (6 × 3 cm2 - 10 × 5 cm2 ). Four flaps were based off of the posterior branch of the STA, while the fifth was based off of the anterior branch. Two donor sites were closed primarily, and three required skin grafts. One patient experienced partial flap necrosis. There were no complete flap losses and no donor site complications. Average follow up was 14.6 months (9-20 months). All patients maintained preoperative level of speech, mastication, and oral continence. CONCLUSIONS: The STAP flap can be based on the anterior or posterior branch of the superficial temporal artery and is a useful regional flap for intraoral defects after tumor resection.

A new distributional record of the black coral Antipathes cf. griggi in Mactan Island, Philippines.

The black coral Antipathes cf. griggi was reported for the first time outside the Hawaiian Archipelago. This short report provides details and short description of the newly recorded species in Mactan Island, Philippines.

Fidaxomicin Compared With Oral Vancomycin for the Treatment of Severe Clostridium difficile-Associated Diarrhea: A Retrospective Review.

Purpose: The most recent published guidelines on Clostridium difficile-associated diarrhea (CDAD) developed by the Infectious Diseases Society of America (IDSA) were released in 2017 and outline its treatment based on severity of the disease and recurrence; however, a clear first-line agent has not been recommended specifically for severe CDAD. Methods: This retrospective chart review was approved by the institutional review board and consisted of three community hospitals and one academic medical center. To be included, patients need to meet criteria for severe CDAD and receive at least 72 hours of therapy. Patients received either oral vancomycin or fidaxomicin, in addition to other therapies for CDAD, and differences in outcomes such as cost obtained from a common charge center, rates of recurrence, time to recurrence as measured at time of positive to negative polymerase chain reaction (PCR) test, and mortality were assessed. Results: Of the 147 patients, 74 patients received fidaxomicin and 73 patients received oral vancomycin. The average hospitalization cost for patients receiving fidaxomicin was $129,338.69 and for patients receiving vancomycin was $153,563.81 (P = .26). Recurrence rates were lower with fidaxomicin compared with vancomycin (6.8% vs 17.6%; P = .047), and time to recurrence was longer with fidaxomicin versus vancomycin, but not statistically significant (96.8 ± 45.9 days vs 63.2 ± 66.9 days; P = .321). Mortality, length of stay in the intensive care unit, and overall length of stay were similar between the two therapies. Conclusions: In the treatment of severe CDAD, recurrence rates were lower and time to recurrence was higher with fidaxomicin compared with oral vancomycin. A clear financial benefit has yet to translate from these known findings.

Revealing architectural order with quantitative label-free imaging and deep learning.

We report quantitative label-free imaging with phase and polarization (QLIPP) for simultaneous measurement of density, anisotropy, and orientation of structures in unlabeled live cells and tissue slices. We combine QLIPP with deep neural networks to predict fluorescence images of diverse cell and tissue structures. QLIPP images reveal anatomical regions and axon tract orientation in prenatal human brain tissue sections that are not visible using brightfield imaging. We report a variant of U-Net architecture, multi-channel 2.5D U-Net, for computationally efficient prediction of fluorescence images in three dimensions and over large fields of view. Further, we develop data normalization methods for accurate prediction of myelin distribution over large brain regions. We show that experimental defects in labeling the human tissue can be rescued with quantitative label-free imaging and neural network model. We anticipate that the proposed method will enable new studies of architectural order at spatial scales ranging from organelles to tissue.

Microscopy is central to biological research and has enabled scientist to study the structure and dynamics of cells and their components within. Often, fluorescent dyes or trackers are used that can be detected under the microscope. However, this procedure can sometimes interfere with the biological processes being studied. Now, Guo, Yeh, Folkesson et al. have developed a new approach to examine structures within tissues and cells without the need for a fluorescent label. The technique, called QLIPP, uses the phase and polarization of the light passing through the sample to get information about its makeup. A computational model was used to decode the characteristics of the light and to provide information about the density and orientation of molecules in live cells and brain tissue samples of mice and human. This way, Guo et al. were able to reveal details that conventional microscopy would have missed. Then, a type of machine learning, known as 'deep learning', was used to translate the density and orientation images into fluorescence images, which enabled the researchers to predict specific structures in human brain tissue sections. QLIPP can be added as a module to a microscope and its software is available open source. Guo et al. hope that this approach can be used across many fields of biology, for example, to map the connectivity of nerve cells in the human brain or to identify how cells respond to infection. However, further work in automating other aspects, such as sample preparation and analysis, will be needed to realize the full benefits.

Negative Thermal Expansion in the Plateau State of a Magnetically Frustrated Spinel.

We report on negative thermal expansion (NTE) in the high-field, half-magnetization plateau phase of the frustrated magnetic insulator CdCr_{2}O_{4}. Using dilatometry, we precisely map the phase diagram at fields of up to 30 T and identify a strong NTE associated with the collinear half-magnetization plateau for B>27 T. The resulting phase diagram is compared with a microscopic theory for spin-lattice coupling, and the origin of the NTE is identified as a large negative change in magnetization with temperature, coming from a nearly localized band of spin excitations in the plateau phase. These results provide useful guidelines for the discovery of new NTE materials.

Chemical capture of wild swamp buffalo (Bubalus bubalis) in tropical northern Australia using thiafentanil, etorphine and azaperone combinations.

BACKGROUND: Studying wild animals in situ is fundamental to collecting baseline information, but generally they need to be immobilised for examination, sampling, marking and/or equipping with tracking apparatus. Capturing wild animals is inherently risky and there is a need for immobilisation methods that are safe for both the animals and researchers. METHODS: A total of 16 free-ranging swamp buffalo (Bubalus bubalis) were chemically captured by dart for the application of satellite tracking collars in tropical northern Australia; 7 animals were anesthetised with a thiafentanil-etorphine-azaperone (TEA) combination and 9 animals with a thiafentanil-azaperone (TA) combination. Anaesthesia was reversed with intravenous naltrexone. Mean dosages of etorphine and thiafentanil for animals in the TEA group were 0.01 mg/kg of each drug and mean dosage of thiafentanil for animals in the TA group was 0.02 mg/kg. Total dose per animal of azaperone and naltrexone was 80 mg and 150 mg, respectively. Anaesthetic monitoring was by physical observation of physiological variables, pulse oximetry and capnography. Blood laboratory parameters including creatine kinase (CK), aspartate transaminase (AST), serum bicarbonate and anion gap were measured. RESULTS: All subject animals recovered well from anaesthesia despite the occurrence of subclinical acidosis in some patients. There was no significant difference between the treatment groups. Conversely, chase time had an adverse effect on body temperature, irrespective of the anaesthetic combination used. CONCLUSIONS: Thiafentanil and azaperone, with or without etorphine, delivered rapid safe, effective, reversible field anaesthesia in healthy swamp buffalo.

Multi-disciplinary approach to perioperative risk assessment and post-transplant management for liver transplantation in a patient at risk for Brugada syndrome.

Brugada syndrome, an autosomal dominant genetic disorder, is characterised by abnormal electrocardiogram findings and increased risk of ventricular tachyarrhythmias and sudden cardiac death. Our report describes the multi-disciplinary perioperative management of a 28-year-old patient presenting to the Duke Transplant Center with a familial sodium channel gene SCN51 mutation concerning Brugada syndrome. We discuss the preparatory work-up, medication review and appropriate post-surgical follow-up for patients undergoing liver transplant surgery with cardiac monitoring.

An ultra-compact low temperature scanning probe microscope for magnetic fields above 30 T.

We present the design of a highly compact high field scanning probe microscope (HF-SPM) for operation at cryogenic temperatures in an extremely high magnetic field, provided by a water-cooled Bitter magnet able to reach 38 T. The HF-SPM is 14 mm in diameter: an Attocube nano-positioner controls the coarse approach of a piezoresistive atomic force microscopy cantilever to a scanned sample. The Bitter magnet constitutes an extreme environment for scanning probe microscopy (SPM) due to the high level of vibrational noise; the Bitter magnet noise at frequencies up to 300 kHz is characterized, and noise mitigation methods are described. The performance of the HF-SPM is demonstrated by topographic imaging and noise measurements at up to 30 T. Additionally, the use of the SPM as a three-dimensional dilatometer for magnetostriction measurements is demonstrated via measurements on a magnetically frustrated spinel sample.

A preliminary trial of an online dissonance-based eating disorder intervention.

OBJECTIVE: We conducted a controlled randomized preliminary trial of an expanded online version of the Body Project (n = 46) compared to an assessment-only control condition (n = 36) via a longitudinal design (baseline, postintervention, 2-month follow-up) in a community sample of women (N = 82) with clinical (n = 53) and subclinical (n = 29) eating disorder symptoms. METHOD: The traditional content of the Body Project was modified to include verbal, written, and behavioral exercises designed to dissuade objectification and maladaptive social comparison and adapted to an online format. Body dissatisfaction, self-esteem, self-objectification, thin-ideal internalization, maladaptive social comparison, trait anxiety, positive affect, negative affect, and eating disorder symptomatology were evaluated in the control and the online expanded Body Project condition at baseline, postintervention, and 2-month follow-up. RESULTS: A 2 (condition: online expanded Body Project, control) × 3 (time: baseline, postintervention, 2-month follow-up) mixed factorial multivariate analysis of variance (MANOVA) was conducted to examine statistically significant group differences. As predicted, results indicated a statistically significant condition × time interaction. CONCLUSIONS: Participants in the expanded online Body Project condition showed significant reductions in eating disorder symptoms and several associated psychological risk correlates from baseline to postintervention and follow-up; contrary to predictions, eating disorder symptoms and risk correlates were not significantly lower in the online expanded Body Project condition compared to the waitlist control condition at postintervention or 2-month follow-up.