Long-Term Tissue Preservation at Ambient Temperature for Post-Mass Fatality Incident DNA-Based Victim Identification.

In a mass fatality incident (MFI), effective preservation of tissue samples is the cornerstone for downstream DNA-based identification of victims. This is commonly achieved through freezing of tissue samples excised from bodies/fragmented remains which may be buried or stored in refrigerated containers. This may, however, not be possible depending on the nature of the MFI; in particular, during armed conflict/war where extended periods of electrical outages would be expected. The present study compared the effectiveness of long-term tissue preservation at ambient temperatures using two commercial products (non-iodized kitchen salt and a 40% alcoholic beverage) against a chemical preservative (Allprotect™ Tissue Reagent (Qiagen, Germantown, MD, USA)) and freezing at -20 °C. Bovine muscle tissue, used as a proxy for human tissue, was treated with the four preservation methods and sampled at six different time-points over a 24-month period. All four methods were able to preserve the bovine tissue, generally yielding STR-PCR (Short Tandem Repeat-Polymerase Chain Reaction) amplicons > 200 bp in size even at the end of 24 months. Gel electrophoresis, however, indicated that salt was more effective in preserving DNA integrity with high-molecular-weight DNA clearly visible as compared to the low-molecular-weight DNA smears observed in the other methods. This study also proposes a simple process for the rapid and low-cost preservation of tissue samples for long-term storage at ambient temperatures in support of post-incident victim identification efforts.

The Spectrum of Relentless Placoid and Acute Posterior Multifocal Placoid Pigment Epitheliopathy (APMPPE): Multimodal Imaging Analysis.

PURPOSE: To showcase the spectrum between APMPPE and relentless placoid utilizing ultra-widefield imaging findings of a case of acute posterior multifocal placoid pigment epitheliopathy (APMPPE) progressing to relentless placoid chorioretinitis (RPC). METHODS: A 23-year-old Caucasian female presented with worsening vision in both eyes. Clinical examination and multimodal imaging modalities including fundus photos, fundus autofluorescence, fluorescein angiography (FA), indocyanine green angiography (ICGA) and spectral-domain optical coherence tomography and angiography were utilized to diagnose APMPPE and clinically follow the patient. RESULTS: Clinical examination of the patient initially revealed posterior lesions consistent with APMPPE but subsequent multimodal images including ultra-widefield FA and ICGA revealed newer, more peripheral lesions more typical of RPC. CONCLUSION: When compared to standard multimodal imaging, ultra-widefield imaging is an effective tool to delineate nuances between APMPPE and RPC through identification of peripheral lesions, which may be of clinical importance when determining management and therapeutics for patients.

ULTRA-WIDEFIELD IMAGING DETECTION RATE IN IDENTIFYING PERIPHERAL RETINAL TEARS IN SINGLE VERSUS MONTAGE OF PERIPHERAL STEERING.

PURPOSE: To compare the detection rate of orthogonal, directed peripheral steering, and automontaged images with ultra-widefield imaging and the factors influencing the ability to identify retinal breaks. DESIGN: Retrospective cohort study. METHODS: Three hundred and seventy-six treatment-naive eyes (349 patients) that underwent laser retinopexy for retinal breaks between 2015 and 2021 were included. Pretreatment ultra-widefield orthogonal, peripheral steering, and automontage were cross-referenced to scleral-depressed examination to determine whether images successfully visualized all retinal breaks. Total relative retinal area (RRA) visualized was divided by its optic disk area (pixels) to calculate relative retinal area. Potential associations were assessed by linear regression analysis. RESULTS: One hundred and sixty two eyes (154 patients) met inclusion criteria. Orthogonal, peripheral steering, and automontage images showed detection rates of 47.5%, 90.7%, and 80.0%, respectively. Relative retinal area increased from orthogonal versus montage by 34.7% ± 26.5% (mean ± SD), which increased the detection rate by 90.8% ( P = 0.006). In linear probability models, vertical meridian tears decreased probability of identification in orthogonal, peripheral steering, and automontage by -26.6%, -86.2%, and -68.7%, respectively ( P < 0.001), and horizontal meridian tears increased the probability by 62.2%, 92.9%, and 85.5%, respectively, ( P < 0.001). Tears posterior to the equator in orthogonal images increased the probability (91.4%, P < 0.001). Artifacts such as lids/lashes, reflection, and face guard decreased the probability in directed peripheral steering by -28.6%, -50.0%, and -66.7%, respectively, ( P = 0.020, P = 0.049, and P = 0.016). CONCLUSION: Using directed peripheral steering and automontage increases RRA and detection rate of identifying peripheral retinal breaks. Tears in horizontal meridians or posterior to the equator increase the probability of identification. Common ultra-widefield imaging artifacts can significantly limit the probability of identifying retinal tears.

Impact of the iWHELD digital person-centered care program on quality of life, agitation and psychotropic medications in people with dementia living in nursing homes during the COVID-19 pandemic: A randomized controlled trial.

INTRODUCTION: iWHELD is a digital person-centered care program for people with dementia in nursing homes adapted for remote delivery during the COVID-19 pandemic. METHODS: A 16-week two-arm cluster-randomized controlled trial in 149 UK nursing homes compared iWHELD with treatment as usual (TAU). Primary outcome was the overall quality of life with secondary outcomes of agitation and psychotropic use. RESULTS: iWHELD conferred benefit to quality of life on the primary (F = 4.3, p = 0.04) and secondary measures of quality of life (F = 6.45, p = 0.01) and reduced psychotropic medication use (χ2  = 4.08, p = 0.04) with no worsening of agitation. Benefit was seen in participants who contracted COVID-19, those with agitation at baseline, and those taking psychotropic medications. DISCUSSION: iWHELD confers benefits to quality of life and key measures of well-being, can be delivered during the challenging conditions of a pandemic, and should be considered for use alongside any emerging pharmacological treatment for neuropsychiatric symptoms. HIGHLIGHTS: iWHELD is the only remote, digital delivery nursing home training programme for dementia care iWHELD improved quality of life in people with dementia and reduced antipsychotic use without worsening of agitation Residents who contracted Covid-19 during the study also experienced benefits from iWHELD iWHELD offers a valuable, pandemic-safe tool for improving dementia care.

Quadrant Asymmetry in Optical Coherence Tomography Angiography Metrics in Ischemic Versus Non-Ischemic Central Retinal Vein Occlusion Eyes.

Purpose: To determine whether quadrant asymmetry (QA) of optical coherence tomography angiography (OCTA) metrics differs between non-ischemic versus ischemic central retinal vein occlusion (CRVO). Methods: Fifty-eight eyes (21 non-ischemic, 10 ischemic CRVO, and 27 contralateral control eyes) underwent 3 × 3 mm spectral-domain OCTA scans with quantification of the superficial retinal layer vessel length density (VLD) and perfusion density (PD). QA, defined as the maximum-minus-minimum value among four parafoveal Early Treatment Diabetic Retinopathy Study (ETDRS) quadrants, was compared by linear regression including fixed effects for each eye. Results: Mean age was 73.6 ± 11.4 (range 39-88), 73.8 ± 12.4 (range 39-91) and 77.2 ± 9.83, (range 60-88); and QA was 3.46 ± 1.76, 3.14 ± 1.57, and 4.88 ± 2.42 for VLD and 0.072 ± 0.038, 0.062 ± 0.036, and 0.11 ± 0.056 for PD for control, non-ischemic, and ischemic, respectively. QA was significantly higher in ischemic (0.109 ± 0.056) than non-ischemic CRVO eyes (0.062 ± 0.036; P = 0.02) and control eyes for PD (0.072 ± 0.038; P = 0.03). QA was also greater in ischemic (4.875 ± 2.418) than non-ischemic CRVO (3.141 ± 1.572) for VLD (P = 0.04). In terms of identifying which particular quadrant is most affected by ischemia, multivariate regression analysis comparing intra-quadrant effect on the presence of ischemia versus non-ischemia showed no quadrant was significantly affected (P > 0.05 for all quadrants). Conclusions: Ischemic CRVO increases intraeye QA of OCTA metrics when compared to non-ischemic CRVO and control eyes. No specific ETDRS quadrant appears to be more affected. Translational Relevance: This work uses an intraeye method to delineate between ischemic and non-ischemic CRVO by OCTA imaging, overcoming inter-eye variables encountered in clinical care.

Applications of Genomics and Transcriptomics in Precision Medicine for Myopia Control or Prevention.

Myopia is a globally emerging concern accompanied by multiple medical and socio-economic burdens with no well-established causal treatment to control thus far. The study of the genomics and transcriptomics of myopia treatment is crucial to delineate disease pathways and provide valuable insights for the design of precise and effective therapeutics. A strong understanding of altered biochemical pathways and underlying pathogenesis leading to myopia may facilitate early diagnosis and treatment of myopia, ultimately leading to the development of more effective preventive and therapeutic measures. In this review, we summarize current data about the genomics and transcriptomics of myopia in human and animal models. We also discuss the potential applicability of these findings to precision medicine for myopia treatment.

Science of malaria elimination: using knowledge of bottlenecks and enablers from the Malaria Elimination Demonstration Project in Central India for eliminating malaria in the Asia Pacific region.

Malaria poses a major public health challenge in the Asia Pacific. Malaria Elimination Demonstration Project was conducted as a public-private partnership initiative in Mandla between State government, ICMR, and FDEC India. The project employed controls for efficient operational and management decisions. IEC campaigns found crucial in schools and communities. Capacity building of local workers emphasized for better diagnosis and treatment. SOCH mobile app launched for complete digitalization. Better supervision for Indoor Residual Sprays and optimized Long Lasting Insecticidal Nets distribution. Significant malaria cases reduction in Mandla. Insights from MEDP crucial for malaria elimination strategies in other endemic regions of the Asia Pacific.

Correlation of Diabetic Disease Severity to Degree of Quadrant Asymmetry in En Face OCTA Metrics.

Purpose: To determine if diabetic retinopathy (DR) severity affects quadrant asymmetry (QA) of optical coherence tomography angiography (OCTA) metrics differentially. Methods: Ninety eyes (60 patients) with no diabetes mellitus (DM) (n = 39) or varying levels of DR (n = 51) had OCTA images (3 × 3 mm, Cirrus5000) acquired five times and averaged. The vessel length density (VLD) and perfusion density (PD) of the superficial retinal layer (SRL) and deep retinal layer (DRL) were measured. QA was defined as the maximum minus minimum value among four parafoveal Early Treatment Diabetic Retinopathy quadrants, and compared with DR severity by linear regression including fixed effects for each individual and eye. Results: The mean patient age was 55.5 years (range, 24-88 years) and 60% were male. Comparing severe nonproliferative DR or proliferative DR versus no DM/DR eyes, QA was significantly higher for SRL VLD, and PD (+0.67 ± 0.16 and +0.014 ± 0.003; P < 0.001) and DRL VLD, and PD (+1.25 ± 0.16 and +0.032 ± 0.003; P < 0.001). When comparing mild or moderate nonproliferative DR versus no DM/DR, the DRL VLD, and PD were significantly higher (+0.51 ± 0.13 and +0.015 ± 0.003; P < 0.001). For every step increase in DR severity, there was a +0.20 QA for SRL VLD, +0.004 SRL PD, +0.33 DRL VLD and +0.009 DRL PD (P < 0.001). Regression analysis comparing intraquadrant effect on DR severity demonstrated that the superior quadrant was most affected for all OCTA metrics. Conclusions: DR severity affects VLD and PD more asymmetrically across Early Treatment Diabetic Retinopathy quadrants with a linear increase in QA for each worsening level of DR. Individual intraeye metrics such as QA can accurately quantify DR severity without concerns for intereye variabilities that could affect the reproducibility and reliability of OCTA quantification.

Impact of Coronavirus Disease 2019 Restrictions on Retinal Detachment: A Multicenter Experience.

In this retrospective, multicenter study of 261 eyes (259 patients), patients who underwent rhegmatogenous retinal detachment repair during the coronavirus disease 2019 (COVID-19) post-lockdown period experienced an additional 22-day delay, leading to significantly more epiretinal membrane and proliferative vitreoretinopathy and lower single-surgery anatomic success rates. During lockdown, perfluoropropane gas was used more commonly, and pneumatic retinopexy was used more commonly in COVID-19-positive patients.

Oxidative stress in cardiac hypertrophy: From molecular mechanisms to novel therapeutic targets.

When faced with increased workload the heart undergoes remodelling, where it increases its muscle mass in an attempt to preserve normal function. This is referred to as cardiac hypertrophy and if sustained, can lead to impaired contractile function. Experimental evidence supports oxidative stress as a critical inducer of both genetic and acquired forms of cardiac hypertrophy, a finding which is reinforced by elevated levels of circulating oxidative stress markers in patients with cardiac hypertrophy. These observations formed the basis for using antioxidants as a therapeutic means to attenuate cardiac hypertrophy and improve clinical outcomes. However, the use of antioxidant therapies in the clinical setting has been associated with inconsistent results, despite antioxidants having been shown to exert protection in several animal models of cardiac hypertrophy. This has forced us to revaluate the mechanisms, both upstream and downstream of oxidative stress, where recent studies demonstrate that apart from conventional mediators of oxidative stress, metabolic disturbances, mitochondrial dysfunction and inflammation as well as dysregulated autophagy and protein homeostasis contribute to disease pathophysiology through mechanisms involving oxidative stress. Importantly, novel therapeutic targets have been identified to counteract oxidative stress and attenuate cardiac hypertrophy but more interestingly, the repurposing of drugs commonly used to treat metabolic disorders, hypertension, peripheral vascular disease, sleep disorders and arthritis have also been shown to improve cardiac function through suppression of oxidative stress. Here, we review the latest literature on these novel mechanisms and intervention strategies with the aim of better understanding the complexities of oxidative stress for more precise targeted therapeutic approaches to prevent cardiac hypertrophy.

Artificial neural network, predictor variables and sensitivity threshold for DNA methylation-based age prediction using blood samples.

Regression models are often used to predict age of an individual based on methylation patterns. Artificial neural network (ANN) however was recently shown to be more accurate for age prediction. Additionally, the impact of ethnicity and sex on our previous regression model have not been studied. Furthermore, there is currently no age prediction study investigating the lower limit of input DNA at the bisulfite treatment stage prior to pyrosequencing. Herein, we evaluated both regression and ANN models, and the impact of ethnicity and sex on age prediction for 333 local blood samples using three loci on the pyrosequencing platform. Subsequently, we trained a one locus-based ANN model to reduce the amount of DNA used. We demonstrated that the ANN model has a higher accuracy of age prediction than the regression model. Additionally, we showed that ethnicity did not affect age prediction among local Chinese, Malays and Indians. Although the predicted age of males were marginally overestimated, sex did not impact the accuracy of age prediction. Lastly, we present a one locus, dual CpG model using 25 ng of input DNA that is sufficient for forensic age prediction. In conclusion, the two ANN models validated would be useful for age prediction to provide forensic intelligence leads.

Optimization of spermatozoa detection using immunofluorescent staining and laser micro-dissection.

The present study evaluated the use of an immunofluorescence-based assay for the microscopic detection of human spermatozoa, following which the fluorescence-labelled spermatozoa could be excised with a laser micro-dissection system. The Sperm Hy-Liter™ PI kit was able to detect spermatozoa from as little as 20nL of semen. No interference or non-specificity were observed when the kit was used on semen mixed with various body fluids such as blood and urine, as well as when semen was spiked onto different types of fabric. Good results could also be obtained with rectal samples which contain auto-fluorescent fecal materials through the use of dual FITC/PI filters. We also developed a method for concurrent testing of two protein biomarkers of semen (semenogelin and prostate-specific antigen) and detection of spermatozoa. This approach would maximize the evidential value from a single piece of sexual assault exhibit. The results also showed that staining by Sperm Hy-Liter™ PI does not interfere with DNA recovery, facilitating the generation of clear male DNA profiles from dissected spermatozoa, thereby making profile interpretation less complex. In summary, Sperm Hy-Liter™ PI staining was demonstrated to be sensitive, robust and specific.