Age differential response to bevacizumab therapy in choroidal neovascularization in rabbits.

Choroidal neovascularization (CNV) in young rabbits has been shown to have a rapid, robust response after treatment with bevacizumab, an anti-vascular endothelial growth factor (VEGF) medication. This investigation evaluates an age differential response to bevacizumab in older populations of rabbits using multimodal high resolution molecular imaging. Young (4 months old) and life span (14 months old) rabbits were given subretinal injections of Matrigel and VEGF to produce CNV. All CNV rabbit models were then treated with a bevacizumab intravitreal injection. Rabbits were then monitored longitudinally using photoacoustic microscopy (PAM), optical coherence tomography (OCT), color photography, and fluorescence imaging. Chain-like gold nanoparticle clusters (CGNP) conjugated with tripeptide arginylglycylaspartic acid (RGD) was injected intravenously for molecular imaging. Robust CNV developed in both young and old rabbits. After intravitreal bevacizumab injection, fluorescence signals were markedly decreased 90.13% in the young group. In contrast, old rabbit CNV area decreased by only 10.56% post-bevacizumab treatment. OCT images confirmed a rapid decrease of CNV in the young group. CGNPs demonstrated high PAM signal in old rabbits and minimal PAM signal in young rabbits after bevacizumab, indicating CNV regression. There is a significant difference in response to intravitreal bevacizumab treatment between young and old rabbits with CNV which can be monitored with multimodal molecular imaging. Old rabbits demonstrate significant persistent disease activity. This represents the first large eye model of persistent disease activity of CNV and could serve as the foundation for future investigations into the mechanism of persistent disease activity and the development of novel therapies.

DEXMEDETOMIDINE, BUTORPHANOL, AND MIDAZOLAM AS A REVERSIBLE INDUCTION PROTOCOL IN NORTH AMERICAN RIVER OTTERS (LONTRA CANADENSIS).

Captured free-ranging North American river otters (Lontra canadensis) were immobilized for the placement of intra-abdominal radio transmitters in cooperation with the Iowa Department of Natural Resources. Twenty-four otters were induced with dexmedetomidine (0.03 mg/kg, IM), butorphanol (0.2 mg/kg, IM), and midazolam (0.15 mg/kg, IM) combined in one syringe. The otters were maintained on isoflurane during the surgical procedure. Heart rate and rhythm, respiratory rate, rectal temperature, and peripheral capillary oxygen saturation were recorded every 5 min for the duration of the procedures. The otters were reversed with atipamezole (0.3-2 mg/kg, IM), naltrexone (0.6 mg/kg, IM), and flumazenil (0.05 mg/kg, IM). Rapid and smooth induction was seen, with lateral recumbency reached within 6.2 ± 5.6 min. Episodes of resedation were seen in four otters that received 0.3 mg/kg atipamezole so the dose was increased to 1 mg/kg, and no further resedation events were noted. Two fatal complications occurred secondary to suspected respiratory arrest during recovery. This drug protocol provided a smooth and rapid induction in free-ranging river otters, but further research is required to determine the safety of this protocol for river otters in both zoo and free-ranging situations.

High Resolution Multimodal Photoacoustic Microscopy and Optical Coherence Tomography Visualization of Choroidal Vascular Occlusion.

Photoacoustic microscopy is a novel, non-ionizing, non-invasive imaging technology that evaluates tissue absorption of short-pulsed light through the sound waves emitted by the tissue and has numerous biomedical applications. In this study, a custom-built multimodal imaging system, including photoacoustic microscopy (PAM) and optical coherence tomography (OCT), has been developed to evaluate choroidal vascular occlusion (CVO). CVO was performed on three living rabbits using laser photocoagulation. Longitudinal imaging of CVO was obtained using multiple imaging tools such as color fundus photography, fluorescein angiography, indocyanine green angiography (ICGA), OCT, and PAM. PAM images were acquired at different wavelengths, ranging from 532 to 700 nm. The results demonstrate that the CVO was clearly observed on PAM in both two dimensions (2D) and 3D with high resolution longitudinally over 28 days. In addition, the location and margin of the CVO were distinguished from the surrounding choroidal vasculature after the injection of ICG contrast agent. PAM imaging was achieved using a laser energy of approximately 80 nJ, which is about half of the American National Standards Institute safety limit. The proposed imaging technique may provide a potential tool for the evaluation of different chorioretinal vascular disease pathogeneses and other biological studies.

The development of patient suicide post-vention guidelines for psychiatry trainees and supervisors.

OBJECTIVES: Patient suicide is one of the most stressful events for psychiatrists and psychiatry trainees. However, there is often a lack of consistent guidance and support available to clinicians, including post-vention. The aim of this paper is to describe the development of a resource that could support psychiatry trainees following patient suicide. METHODS: Following a literature review, we adapted two US patient suicide post-vention guidelines to reflect local processes and support systems available by consulting a number of key stakeholders in the training programme and district health boards. RESULTS: The first part of the post-vention guidelines included procedural processes such as the serious incident review process and reporting to coroner's office. The second part included a checklist for trainee, supervisor, local training facilitator and director of training according to the following time frame: first 24 hours, first 1-2 weeks and following months. CONCLUSIONS: Post-vention guidelines and teaching about patient suicide and its effects could improve the training experience of psychiatry trainees and facilitate the development of resilience as they progress through training.

Multimodal Imaging-Guided Stem Cell Ocular Treatment.

Stem cell therapies are gaining traction as promising treatments for a variety of degenerative conditions. Both clinical and preclinical studies of regenerative medicine are hampered by the lack of technologies that can evaluate the migration and behavior of stem cells post-transplantation. This study proposes an innovative method to longitudinally image in vivo human-induced pluripotent stem cells differentiated to retinal pigment epithelium (hiPSC-RPE) cells by multimodal photoacoustic microscopy, optical coherence tomography, and fluorescence imaging powered by ultraminiature chain-like gold nanoparticle cluster (GNC) nanosensors. The GNC exhibits an optical absorption peak in the near-infrared regime, and the 7-8 nm size in diameter after disassembly enables renal excretion and improved safety as well as biocompatibility. In a clinically relevant rabbit model, GNC-labeled hiPSC-RPE cells migrated to RPE degeneration areas and regenerated damaged tissues. The hiPSC-RPE cells' distribution and migration were noninvasively, longitudinally monitored for 6 months with exceptional sensitivity and spatial resolution. This advanced platform for cellular imaging has the potential to enhance regenerative cell-based therapies.

Multimodal imaging of laser-induced choroidal neovascularization in pigmented rabbits.

This study aimed to demonstrate longitudinal multimodal imaging of laser photocoagulation-induced choroidal neovascularization (CNV) in pigmented rabbits. Six Dutch Belted pigmented rabbits were treated with 12 laser lesions in each eye at a power of 300 mW with an aerial diameter spot size of 500 µm and pulse duration of 100 ms. CNV progression was monitored over a period of 4 months using different imaging techniques including color fundus photography, fluorescein angiography (FA), photoacoustic microscopy (PAM), and optical coherence tomography (OCT). All treated eyes developed CNV with a success rate of 100%. The margin and morphology of CNV were detected and rendered in three dimensions using PAM and OCT. The CNV was further distinguished from the surrounding melanin and choroidal vessels using FDA-approved indocyanine green dye-enhanced PAM imaging. By obtaining PAM at 700 nm, the location and density of CNV were identified, and the induced PA signal increased up to 59 times. Immunohistochemistry with smooth muscle alpha-actin (αSMA) antibody confirmed the development of CNV. Laser photocoagulation demonstrates a great method to create CNV in pigmented rabbits. The CNV was stable for up to 4 months, and the CNV area was measured from FA images similar to PAM and OCT results. In addition, this study demonstrates that contrast agent-enhanced PAM imaging allows for precise visualization and evaluation of the formation of new blood vessels in a clinically-relevant animal model of CNV. This laser-induced CNV model can provide a unique technique for longitudinal studies of CNV pathogenesis that can be imaged with multimodal imaging.

Successful treatment of 5-fluorouracil toxicosis with hemodialysis.

OBJECTIVE: To describe the successful treatment of lethal dose 5-fluorouracil (5-FU) toxicosis using hemodialysis. CASE SUMMARY: A 4-month-old intact female Golden Retriever was presented to the emergency department after ingesting 20 g of 5% 5-FU cream. The puppy developed refractory seizures and became comatose with uncontrolled tonic-clonic convulsions. Because of the low molecular weight and minimal protein binding of 5-FU, a single hemodialysis treatment was employed for detoxification. The puppy improved clinically posttreatment and was successfully discharged 3 days after admission. Postingestion leukopenia and neutropenia occurred but were responsive to treatment with filgrastim. The puppy is neurologically normal and has no lasting effects 1 year postingestion. NEW OR UNIQUE INFORMATION PROVIDED: To the authors' knowledge, this is the first reported case in veterinary medicine of a potentially fatal 5-FU ingestion that has been treated with intermittent hemodialysis.

Renally Clearable Ultraminiature Chain-Like Gold Nanoparticle Clusters for Multimodal Molecular Imaging of Choroidal Neovascularization.

Currently, available gold nanoparticles (GNPs) typically accumulate in the liver and spleen, leading to concerns for their long-term biosafety. To address this long-standing problem, ultraminiature chain-like gold nanoparticle clusters (GNCs) are developed. Via self-assembly of 7-8 nm GNP monomers, GNCs provide redshifted optical absorption and scattering contrast in the near-infrared window. After disassembly, GNCs turn back to GNPs with a size smaller than the renal glomerular filtration size cutoff, allowing their excretion via urine. A one-month longitudinal study in a rabbit eye model demonstrates that GNCs facilitate multimodal molecular imaging of choroidal neovascularization (CNV) in vivo, non-invasively, with excellent sensitivity and spatial resolution. GNCs targeting αv ß3  integrins enhance photoacoustic and optical coherence tomography (OCT) signals from CNV by 25.3-fold and 150%, respectively. With excellent biosafety and biocompatibility demonstrated, GNCs render a first-of-its-kind nanoplatform for biomedical imaging.

Multimodal In Vivo Imaging of Retinal and Choroidal Vascular Occlusion.

Photoacoustic microscopy (PAM) is an emerging retinal imaging technique that can provide high spatial resolution and high contrast of chorioretinal vessels. PAM is compatible with optical coherence tomography (OCT) and fluorescence imaging, allowing for development of a multimodal imaging system that combines these imaging modalities into one. This study presents a non-invasive, label-free in vivo imaging of retinal and choroidal vascular occlusion using multimodal imaging system, including PAM and OCT. Both retinal vein occlusion (RVO) and choroidal vascular occlusion (CVO) were clearly identified selectively using a spectroscopic PAM imaging. RVO and CVO were created in six rabbits using laser photocoagulation. The dynamic changes of retinal vasculature were observed and evaluated using color fundus photography, fluorescein angiography, OCT, and PAM. The position of RVO and CVO were imaged with different wavelengths ranging from 532 to 600 nm. The data shows that occluded vessels were clearly distinguished from the surrounding retinal vessels on the PAM images. This advanced imaging system is a promising technique for imaging retinal ischemia in preclinical disease models.

Medical students' evaluation of a suicide prevention multimedia resource: A focus group study.

AIMS: A series of podcasts and videos was created to assist medical students with learning about suicide prevention. The aim of this research was to explore medical students experiences of using a suicide prevention learning resource. METHODS: A multimedia repository of learning resources for suicide prevention was designed and created for use across all years of the medical programme at The University of Auckland. Emphasis was placed on ensuring that the resource was culturally safe. The impact of the learning resource was evaluated with a qualitative approach using focus group methodology. Two focus groups were audio-recorded, transcribed and a thematic analysis was conducted employing three cycles of coding. RESULTS: Three themes were identified: perceiving that suicide is complex and sensitive; tailoring knowledge to match students' developmental stage and context; and elements that facilitated interaction with the resource. CONCLUSIONS: Suicide is unsurprisingly a challenging topic for medical students. The students in this study actively engaged with this resource on suicide prevention, which supplemented their core learning of the topic. Early access to resources developed in a culturally safe way within a spiral curriculum empowers students to understand that they have an important contribution to make in preventing suicide. This may prepare them for encountering suicide with peers, family members and in clinical practice.

Treatment of Severe Emotionally Unstable Personality Disorder with Comorbid Ehlers-Danlos Syndrome and Functional Neurological Disorder in an Inpatient Setting: A Case for Specialist Units without Restrictive Interventions.

We present the case of a young woman with an Emotionally Unstable Personality Disorder (EUPD) diagnosis suffering from high-risk self-injurious behaviour. She was also diagnosed with Ehlers-Danlos Syndrome and Functional Neurological Disorder, manifesting as nonepileptic seizures and immobility. Our patient, "A," endured traumatic childhood abuse and became highly dependent on services in her late teens. Recurrent suicide attempts resulted in twenty to thirty acute psychiatric admissions, Intensive Care Unit stays, and multiple failed trials of psychological therapy. Nonepileptic seizures and wheelchair dependency made her "too complex" for many specialist services. She was eventually admitted to Springbank ward in Fulbourn Hospital, Cambridge. The EUPD specialist unit prides itself on evidence-based treatments, shared values, and a least restrictive approach. At discharge, our patient was self-harm free and able to walk unaided and no longer met EUPD diagnostic criteria. We include "A's" personal views on her illness and how Springbank ward facilitated her recovery, together with results from structured clinical outcome measures.

Gold Nanorod Enhanced Photoacoustic Microscopy and Optical Coherence Tomography of Choroidal Neovascularization.

Visualization and evaluation of choroidal neovascularization (CNV) are major challenges to improve treatment outcomes for patients with age-related macular degeneration (AMD). Limitations of current imaging techniques include the limited penetration depth, spatial resolution, and sensitivity and difficulty visualizing CNV from the healthy microvasculature. In this study, a custom-built multimodal photoacoustic microscopy (PAM) and optical coherence tomography (OCT) system was developed to distinguish the margin of CNV in living rabbits with the assistance of functionalized gold nanorods conjugating with RGD ligands (GNR-RGD). Intravenous administration of GNR-RGD into rabbits in a CNV model resulted in signal enhancements of 27.2-fold in PAM and 171.4% in OCT. This molecular imaging technique of contrast-enhanced PAM and OCT is a promising tool for the precise imaging of CNV as well as the evaluation of the pathophysiology in vivo without destruction of tissue.

Sonographic Diagnosis of COVID-19: A Review of Image Processing for Lung Ultrasound.

The sustained increase in new cases of COVID-19 across the world and potential for subsequent outbreaks call for new tools to assist health professionals with early diagnosis and patient monitoring. Growing evidence around the world is showing that lung ultrasound examination can detect manifestations of COVID-19 infection. Ultrasound imaging has several characteristics that make it ideally suited for routine use: small hand-held systems can be contained inside a protective sheath, making it easier to disinfect than X-ray or computed tomography equipment; lung ultrasound allows triage of patients in long term care homes, tents or other areas outside of the hospital where other imaging modalities are not available; and it can determine lung involvement during the early phases of the disease and monitor affected patients at bedside on a daily basis. However, some challenges still remain with routine use of lung ultrasound. Namely, current examination practices and image interpretation are quite challenging, especially for unspecialized personnel. This paper reviews how lung ultrasound (LUS) imaging can be used for COVID-19 diagnosis and explores different image processing methods that have the potential to detect manifestations of COVID-19 in LUS images. Then, the paper reviews how general lung ultrasound examinations are performed before addressing how COVID-19 manifests itself in the images. This will provide the basis to study contemporary methods for both segmentation and classification of lung ultrasound images. The paper concludes with a discussion regarding practical considerations of lung ultrasound image processing use and draws parallels between different methods to allow researchers to decide which particular method may be best considering their needs. With the deficit of trained sonographers who are working to diagnose the thousands of people afflicted by COVID-19, a partially or totally automated lung ultrasound detection and diagnosis tool would be a major asset to fight the pandemic at the front lines.

Indocyanine green-enhanced multimodal photoacoustic microscopy and optical coherence tomography molecular imaging of choroidal neovascularization.

Photoacoustic microscopy (PAM) has great potential for visualization of the microvasculature with high spatial resolution and contrast. Early detection and differentiation of newly developed blood vessels named choroidal neovascularization (CNV) from normal vasculature remains a challenge in ophthalmology. Exogenous contrast agents can assist with improving PAM sensitivity, leading to differentiation of CNV. Here, an FDA-approved indocyanine green (ICG) was utilized as a PAM contrast agent. ICG was conjugated with RGD peptides, allowing the ICG to bind to the integrin expressed in CNV. Molecular PAM imaging showed that ICG-RGD can target CNV for up to 5 days post intravenous administration in living rabbits with a model of CNV. The PAM image sensitivity and image contrast were significantly enhanced by 15-fold at 24 h post-injection. Overall, the presented approach demonstrates the possibility of targeted ICG to be employed in PAM molecular imaging, allowing more precise evaluation of neovascularization.

Chain-like gold nanoparticle clusters for multimodal photoacoustic microscopy and optical coherence tomography enhanced molecular imaging.

Colloidal gold nanoparticles (GNPs) serve as promising contrast agents in photoacoustic (PA) imaging, yet their utility is limited due to their absorption peak in the visible window overlapping with that of hemoglobin. To overcome such limitation, this report describes an ultrapure chain-like gold nanoparticle (CGNP) clusters with a redshift peak wavelength at 650 nm. The synthesized CGNP show an excellent biocompatibility and photostability. These nanoparticles are conjugated with arginine-glycine-aspartic acid (RGD) peptides (CGNP clusters-RGD) and validated in 12 living rabbits to perform multimodal photoacoustic microscopy (PAM) and optical coherence tomography (OCT) for visualization of newly developed blood vessels in the sub-retinal pigment epithelium (RPE) space of the retina, named choroidal neovascularization (CNV). The PAM system can achieve a 3D PAM image via a raster scan of 256 × 256 pixels within a time duration of 65 s. Intravenous injection of CGNP clusters-RGD bound to CNV and resulted in up to a 17-fold increase in PAM signal and 176% increase in OCT signal. Histology indicates that CGNP clusters could disassemble, which may facilitate its clearance from the body.

In Vivo Subretinal ARPE-19 Cell Tracking Using Indocyanine Green Contrast-Enhanced Multimodality Photoacoustic Microscopy, Optical Coherence Tomography, and Fluorescence Imaging for Regenerative Medicine.

Purpose: Cell-based regenerative therapies are being investigated as a novel treatment method to treat currently incurable eye diseases, such as geographic atrophy in macular degeneration. Photoacoustic imaging is a promising technology which can visualize transplanted stem cells in vivo longitudinally over time in the retina. In this study, a US Food and Drug Administration (FDA)-approved indocyanine green (ICG) contrast agent is used for labeling and tracking cell distribution and viability using multimodal photoacoustic microscopy (PAM), optical coherence tomography (OCT), and fluorescence imaging. Methods: Twelve rabbits (2.4-3.4 kg weight, 2-4 months old) were used in the study. Human retinal pigment epithelial cells (ARPE-19) were labeled with ICG dye and transplanted in the subretinal space in the rabbits. Longitudinal PAM, OCT, and fluorescence imaging was performed for up to 28 days following subretinal administration of ARPE-19 cells. Results: Cell migration location, viability, and cell layer thickness were clearly recognized and determined from the fluorescence, OCT, and PAM signal. The in vivo results demonstrated that fluorescence signal increased 37-fold and PAM signal enhanced 20-fold post transplantation. Conclusions: This study demonstrates that ICG-assisted PAM, OCT, and fluorescence imaging can provide a unique platform for tracking ARPE-19 cells longitudinally with high resolution and high image contrast. Translational Relevance: Multimodal PAM, OCT, and fluorescence in vivo imaging with ICG can improve our understanding of the fate, distribution, and function of regenerative cell therapies over time nondestructively.

Long-Term, Noninvasive In Vivo Tracking of Progenitor Cells Using Multimodality Photoacoustic, Optical Coherence Tomography, and Fluorescence Imaging.

Stem cell regenerative medicine therapies have emerged as promising treatments for currently incurable diseases. A remaining challenge for cell therapies is the ability to track the migration and distribution of the transplanted cells in a long-term, noninvasive manner in vivo to assess their efficacy. This study develops a noninvasive, and high spatial resolution photoacoustic microscopy (PAM) and optical coherence tomography (OCT) imaging system for in vivo tracking of subretinally injected progenitor human retinal pigment epithelium cells (ARPE-19) labeled with chainlike gold nanoparticle (CGNP) clusters in RPE damage. CGNP provided significant PAM, OCT, and fluorescence signals to selectively track the migration of ARPE-19 cells in living rabbit eyes for 3 months. PAM and OCT imaging allow accurate anatomical information to determine the exact retinal layer in which the transplanted ARPE-19 cells are located which was confirmed by histology. This presents an efficient and advanced technology to visualize fundamental biological processes of cell therapies in complex in vivo environments in real time.

Plasmonic Gold Nanostar-Enhanced Multimodal Photoacoustic Microscopy and Optical Coherence Tomography Molecular Imaging To Evaluate Choroidal Neovascularization.

Although photoacoustic microscopy (PAM) and optical coherence tomography (OCT) allow visualization of the retinal microvasculature, distinguishing early neovascularization from adjacent vessels remains challenging. Herein, gold nanostars (GNSs) functionalized with an RGD peptide were utilized as multimodality contrast agents for both PAM and OCT. GNSs have great absorption and scattering characteristics in the near-infrared region where most vasculature and tissue generates a less intrinsic photoacoustic signal while having a small size, excellent biocompatibility in vivo, and great photostability under nanosecond pulsed laser illumination. This enabled visualization and differentiation of individual microvasculature in vivo using multimodal PAM and OCT imaging. Detailed three-dimensional imaging of GNSs was achieved in an important choroidal neovascularization model in living rabbits. Through the administration of GNSs, PA contrast increased up to 17-fold and OCT intensities increased 167%. This advanced molecular-imaging platform with GNSs provides a unique tool for detailed mapping of the pathogenesis of the microvasculature.

Brain aging modulates the neuroprotective effects of estrogen on selective aspects of cognition in women: a critical review.

Although there is now a substantial literature on the putative neuroprotective effects of estrogen on cognitive functioning in postmenopausal women, it is replete with inconsistencies. The critical period hypothesis, posited several years ago, attempts to account for the discrepancies in this literature by positing that estrogen treatment (ET) will protect aspects of cognition in older women only when treatment is initiated soon after the menopause. Indeed, evidence from basic neuroscience and from the animal and human literature reviewed herein provides compelling support for the critical period hypothesis. Although it is not known with certainty why estrogen does not protect cognition and may even cause harm when administered to women over the age of 65years, it is likely that the events that characterize brain aging, such as a reduction in brain volume and in neuronal size, alterations in neurotransmitter systems, and a decrease in dendritic spine numbers, form an unfavorable background that precludes a neuroprotective effects of exogenous estrogen on the brain. Other factors that have likely contributed to the discrepancies in the estrogen-cognition literature include differences in the estrogen compounds used, their route of administration, cyclic versus continuous regimens, and the concomitant use of progestins. This critical analysis attempts to define conditions under which ET may protect aspects of cognition in aging women while also considering the cost/benefit ratio for the treatment of women aged 50-59years. Suggestions for specific future research questions are also addressed.