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Microphthalmia-associated transcription factor (MITF) is a master regulator of melanocyte function, development and plays a significant role in melanoma pathogenesis. MITF genomic amplification promotes melanoma development, and it can facilitate resistance to multiple therapies. Here, we show that MITF regulates a global antioxidant program that increases survival of melanoma cell lines by protecting the cells from reactive oxygen species (ROS)-induced damage. In addition, this redox program is correlated with MITF expression in human melanoma cell lines and patient-derived melanoma samples. Using a zebrafish melanoma model, we show that MITF decreases ROS-mediated DNA damage in vivo. Some of the MITF target genes involved, such as IDH1 and NNT, are regulated through direct MITF binding to canonical enhancer box (E-BOX) sequences proximal to their promoters. Utilizing functional experiments, we demonstrate the role of MITF and its target genes in reducing cytosolic and mitochondrial ROS. Collectively, our data identify MITF as a significant driver of the cellular antioxidant state.

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A case of septic pulmonary thromboembolism arising from thrombophlebitis of the prostatic venous plexus associated with long-term urinary catheterisation in a 51-year-old man is reported. Despite a previous autopsy having been conducted in the country where he had been resident overseas, a re-examination showed histological evidence of mild patchy chronic prostatitis with a florid, focally purulent, thrombophlebitis of the periprostatic venous plexus with abscess formation and evidence of bacterial overgrowth. Corresponding microscopy of the lungs showed septic microthromboemboli within small pulmonary arteries with variable degrees of necrotising acute inflammation and thrombosis. Death was not due to 'acute pulmonary oedema' as had been originally certified but to septic thromboembolism. This case demonstrates the need to carefully evaluate the prostatic venous plexus at autopsy, and also the type of problems that may arise at the time of the re-examination of repatriated remains.

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The soybean cyst nematode (SCN; Heterodera glycines) facilitates infection by secreting a repertoire of effector proteins into host cells to establish a permanent feeding site composed of a syncytium of root cells. Among the diverse proteins secreted by the nematode, we were specifically interested in identifying proteases to pursue our goal of engineering decoy substrates that elicit an immune response when cleaved by an SCN protease. We identified a cysteine protease that we named Cysteine Protease 1 (CPR1), which was predicted to be a secreted effector based on transcriptomic data obtained from SCN esophageal gland cells, presence of a signal peptide, and lack of transmembrane domains. CPR1 is conserved in all isolates of SCN sequenced to date, suggesting it is critical for virulence. Transient expression of CPR1 in Nicotiana benthamiana leaves suppressed cell death induced by a constitutively active nucleotide binding leucine-rich repeat protein, RPS5, indicating that CPR1 inhibits effector-triggered immunity. CPR1 localizes in part to the mitochondria when expressed in planta. Proximity-based labeling in transgenic soybean roots, co-immunoprecipitation, and cleavage assays identified a branched-chain amino acid aminotransferase from soybean (GmBCAT1) as a substrate of CPR1. Consistent with this, GmBCAT1 also localizes to mitochondria. Silencing of the CPR1 transcript in the nematode reduced penetration frequency in soybean roots while the expression of CPR1 in soybean roots enhanced susceptibility. Our data demonstrates that CPR1 is a conserved effector protease with a direct target in soybean roots, highlighting it as a promising candidate for decoy engineering.

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As a part of a study of suicide in children aged 17 years and under in three centers-Hennepin County in the United States (US), Auckland in New Zealand (NZ), and South Australia in Australia (AUS) from 2008 to 2017 it was decided to characterize potential risk factors and to determine whether these differed by jurisdiction. Reviewed data included a history of psychiatric illness, symptoms prior to suicide, events preceding suicide, previous suicidal ideation or suicide attempts, and communication of suicidal intent. The most common events preceding suicide were arguments with family/friends and relationship issues; in addition depression with or without expressed suicidal ideation, self-harming behavior, sadness, distress, drug/substance abuse, and anorexia were documented. Suicidal intent was on occasion communicated via technological means. In 79.5% of cases in South Australia decedents had a previously diagnosed psychiatric illness, with 62% in Hennepin County. This compared to a much lower proportion of cases in Auckland (23.8%). Whether this reflects more limited access to psychiatric services or a reluctance to seek support and therapy in Auckland is unclear. It does, however, demonstrate that risk factors for child suicide are not uniform among communities and so extrapolation of data from one area to another may not be appropriate. Disturbingly parents/carers were not aware of the decedent's suicidal intent in 84-87.2% of cases.

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ABSTRACT: Helicopters are used worldwide in a range of commercial and private industries and are particularly useful in northern Australia due to the sparse population scattered over large distances with remote regions difficult to access by any other means of transport. Nine civilian helicopter-related fatalities were reported to the Northern Territory coroner between January 2004 and December 2023. The victims were all male (age range, 34-74 years; mean, 52 years). All fatal crashes occurred in remote areas, the majority on cattle stations. Pilots were the sole occupants in 5/9 incidents; in 4 incidents, there were injured survivors. All fatal incidents involved Robinson piston engine helicopters (Robinson R44 models in 2 incidents and Robinson R22 models in the remaining 7). Scene investigations, postmortem examinations, and ancillary investigations were frequently hampered by decomposition of the remains, resulting from difficult search operations and delayed location, retrieval, and storage. Head injuries were present in all cases with chest and spinal injuries in more than half. Integrated assessment of injuries with toxicological and aviation investigations facilitated understanding of the crash dynamics.

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Objective: Aripiprazole lauroxil (AL), a long-acting injectable antipsychotic, has 2 initiation options: 1-day (AL NanoCrystal Dispersion [ALNCD] injection plus 30 mg oral aripiprazole on day 1 only) and 21-day (15 mg oral aripiprazole for 21 days). This post hoc analysis assessed the safety and tolerability of both initiation approaches.Methods: We analyzed data from the first 4 weeks of 2 AL studies, one using the 1-day initiation regimen (conducted between November 2017 and March 2019) and the other using the 21-day initiation regimen (conducted between December 2011 and March 2014). Outcomes of interest during the matched 4-week period included the likelihood of adverse events (AEs), including those associated with discontinuation, rated as serious, or of special interest (injection site reactions [ISRs] and akathisia).Results: The 1-day (n = 99) and 21-day (n = 415) initiation regimens had comparable rates of AEs (57.6% and 52.0%, respectively; most were mild), serious AEs (2.0% and 1.4%), and AEs leading to discontinuation (4.0% and 3.1%). The incidence of ISRs was 11.1% after the ALNCD injection (day 1) in the 1-day initiation regimen. ISR rates for the AL starting doses were 9.2% for the 1-day regimen (AL 1064 mg on day 8) and 3.9% for the 21-day regimen (AL 441 mg/882 mg on day 1). Rates of akathisia were 9.1% and 11.1% for the 1-day and 21-day regimens, respectively. One patient discontinued because of an ISR in the 21-day study, and 2 patients in the 21-day study discontinued because of akathisia. Mean changes from baseline in week 4 Positive and Negative Syndrome Scale total scores were -17.4 (1-day) and -19.5 (21-day).Conclusions: Four-week safety and tolerability were similar following the initiation of AL with either the 1-day or 21-day regimen, supporting the utility of both initiation regimens. Engaging patients in discussions regarding options for initiating AL may help facilitate shared decision-making and personalization of treatment for patients with schizophrenia.Trial Registration: ClinicalTrials.gov identifiers: NCT03345979 and NCT01469039.

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Reactions of bis(benzene)chromium (Bz2Cr) and ozone (O3) were studied using low-temperature argon matrix-isolation infrared spectroscopy with supporting DFT calculations. When Bz2Cr and O3 were co-deposited, they reacted upon matrix deposition to produce two new prominent peaks in the infrared spectrum at 431 cm-1 and 792 cm-1. These peaks increased upon annealing the matrix to 35 K and decreased upon UV irradiation at λ = 254 nm. The oxygen-18 and mixed oxygen-16,18 isotopic shift pattern of the peak at 792 cm-1 is consistent with the antisymmetric stretch of a symmetric ozonide species. DFT calculations of many possible ozonide products of this reaction were made. The formation of a hydrogen ozonide (H2O3) best fits the original peaks and the oxygen-18 isotope shift pattern. Energy considerations lead to the conclusion that the chromium-containing product of this reaction is the coupled product benzene-chromium-biphenyl-chromium-benzene (BzCrBPCrBz). 2Bz2Cr+O3→H2O3+BzCrBPCrBz, ∆Ecalc=-52.13kcal/mol.

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Introduction: The accuracy of musculoskeletal models and simulations as methods for predicting muscle functional outputs is always improving. However, even the most complex models contain various assumptions and simplifications in how muscle force generation is simulated. One common example is the application of a generalised ("generic") force-velocity relationship, derived from a limited data set to each muscle within a model, uniformly across all muscles irrespective of whether those muscles have "fast" or "slow" contractile properties. Methods: Using a previously built and validated musculoskeletal model and simulation of trotting in the mouse hindlimb, this work examines the predicted functional impact of applying muscle-specific force-velocity properties to typically fast (extensor digitorum longus; EDL) and slow-contracting (soleus; SOL) muscles. Results: Using "real" data led to EDL producing more positive work and acting significantly more spring-like, and soleus producing more negative work and acting more brake-like in function compared to muscles modelled using "generic" force-velocity data. Extrapolating these force-velocity properties to other muscles considered "fast" or "slow" also substantially impacted their predicted function. Importantly, this also further impacted EDL and SOL function beyond that seen when changing only their properties alone, to a point where they show an improved match to ex vivo experimental data. Discussion: These data suggest that further improvements to how musculoskeletal models and simulations predict muscle function should include the use of different values defining their force-velocity relationship depending on their fibre-type composition.

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Drinking water security in Puerto Rico (PR) is increasingly challenged by both regulated and emerging anthropogenic contaminants, which was exacerbated by the Hurricane Maria (HM) due to impaired regional water cycle and damaged water infrastructure. Leveraging the NIEHS PROTECT (Puerto Rico Testsite for Exploring Contamination Threats) cohort, this study assessed the long-term tap water (TW) quality changes from March 2018 to November 2018 after HM in PR, by innovatively integrating two different effect-based quantitative toxicity assays with a targeted analysis of 200 organic and 22 inorganic pollutants. Post-hurricane PR TW quality showed recovery after >6-month period as indicated by the decreased number of contaminants showing elevated average concentrations relative to pre-hurricane samples, with significant difference of both chemical and toxicity levels between northern and southern PR. Molecular toxicity profiling and correlation revealed that the HM-accelerated releases of certain pesticides and PPCPs could exert increased cellular oxidative and/or AhR (aryl hydrocarbon receptor)-mediated activities that may persist for more than six months after HM. Maximum cumulative ratio and adverse outcome pathway (AOP) assessment identified the top ranked detected TW contaminants (Cu, Sr, V, perfluorooctanoic acid) that potentially associated with different adverse health effects such as inflammation, impaired reproductive systems, cancers/tumors, and/or organ toxicity. These insights can be incorporated into the regulatory framework for post-disaster risk assessment, guiding water quality control and management for public health protection.

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Autolytic and putrefactive processes can cause considerable alterations to soft tissues and internal organs that may complicate forensic assessments. An overview was undertaken of the range of taphonomonic changes and processes that may result from postmortem putrefactive gas accumulation. The most commonly encountered phenomenon was purging of putrefactive fluids from the nose and mouth that was on occasion confused with bleeding from antemortem trauma. Much less common was putrefactive 'rigor mortis' where the limbs extend due to the accumulation of soft tissue and subcutaneous gas. This may sometimes be associated with alteration of the position of a body suggesting that it had been deliberately moved. Distension and stretching of the skin and subcutaneous tissues may cause recently sutured surgical incisions to dehisce, raising the possibility of inflicted incised wounds. Raised intra-abdominal pressures may cause diaphragmatic herniation of small intestine and has been associated with so-called 'coffin birth' where a fetus is expelled from the uterus after death due to pressure on the fundus. Gas accumulation on postmortem computed tomography examination may be confused with air embolism or the effects of trauma. All of these changes are the result of anaerobic bacterial action generating gases such as methane, carbon dioxide and hydrogen sulphide resulting in pressure gradients.

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PURPOSE: To describe the prevalence and antibiotic resistance profiles of Pseudomonas aeruginosa isolated from the Asia Cornea Society Infectious Keratitis Study (ACSIKS). METHODS: All bacterial isolates from ACSIKS underwent repeat microbiological identification in a central repository in Singapore. Minimum inhibitory concentration (MIC) determination was conducted for isolates of P. aeruginosa against thirteen antibiotics from 6 different classes, and categorized based on Clinical Laboratory Standard Institutes' reference ranges. The percentage rates of resistance (non-susceptibility) to each antibiotic included isolates of both intermediate and complete resistance. Multi-drug resistance (MDR) was defined as non-susceptibility to at least one agent in three or more antimicrobial classes. RESULTS: Of the 1493 unique bacterial specimens obtained from ACSIKS, 319 isolates were of P. aeruginosa. The majority of isolates were from centers in India (n = 118, 37%), Singapore (n = 90, 28.2%), Hong Kong (n = 31, 9.7%) and Thailand (n = 30, 9.4%). The cumulative antibiotic resistance rate was the greatest for polymyxin B (100%), ciprofloxacin (17.6%) and moxifloxacin (16.9%), and lowest for cefepime (11.6%) and amikacin (13.5%). Isolates from India demonstrated the highest antibiotic resistance rates of all the centers, and included moxifloxacin (47.5%) and ciprofloxacin (39.8%). Forty-eight of the 59 MDR isolates also originated from India. Antibiotic resistance rates were significantly lower in the other ACSIKS centers, and were typically less than 10%. CONCLUSIONS: The antibiotic resistance profiles of P. aeruginosa varied between different countries. While it was low for most countries, substantial antibiotic resistance and a significant number of multi-drug resistant isolates were noted in the centers from India.

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Foot-and-mouth disease (FMD) is a severe and extremely contagious viral disease of cloven-hoofed domestic and wild animals, which leads to serious economic losses to the livestock industry globally. FMD is caused by the FMD virus (FMDV), a positive-strand RNA virus that belongs to the genus Aphthovirus, within the family Picornaviridae. Early detection and characterization of FMDV strains are key factors to control new outbreaks and prevent the spread of the disease. Here, we describe a direct RNA sequencing method using Oxford Nanopore Technology (ONT) Flongle flow cells on MinION Mk1C (or GridION) to characterize FMDV. This is a rapid, low cost, and easily deployed point of care (POC) method for a near real-time characterization of FMDV in endemic areas or outbreak investigation sites. Key features • Saves ~35 min of the original protocol time by omitting the reverse transcription step and lowers the costs of reagents and consumables. • Replaces the GridION flow cell from the original protocol with the Flongle, which saves ~90% on the flow cell cost. • Combines the NGS benchwork with a modified version of our African swine fever virus (ASFV) fast analysis pipeline to achieve FMDV characterization within minutes. Graphical overview Schematic of direct RNA sequencing of foot-and-mouth disease virus (FMDV) process, which takes ~50 min from extracted RNA to final loading, modified from the ONT SQK-RNA002 protocol (Version: DRS_9080_v2_revO_14Aug2019).

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Opioid abuse is a leading cause of drug-related morbidity and mortality worldwide. It has been suggested that the age of opiate users in Australia is rising. To evaluate this further in a local population, toxicology and pathology case files from Forensic Science SA, Adelaide, South Australia, were examined for all cases with lethal opioid levels from 2000 to 2019 (n = 499; M:F 2.3:1; age range 18-91 years, median age 42 years). The median age of opiate deaths increased significantly by approximately 16 years (p = 0.007, R2 = 0.34) with a significant increase in total deaths in the 45-54 years and 55-64 years age groups (p = 0.009, R2 = 0.32) (p = < 0.001, R2 = 0.54). Deaths due to heroin overdose showed the lowest median age (39 years, n = 184), with deaths from tramadol toxicity having the highest (50.5 years, n = 32). Recent changes in the demographic profile of opioid users in cases of lethal overdose involve an aging population. Forensic and clinical practitioners should be aware of significant opioid abuse in certain individuals at older ages as this raises the possibility that this may exacerbate the effects of age-related chronic diseases in this group and/or contribute to fatalities.

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A 39-year-old woman was found lying in the rear of a car at her home address. A smell of rotten eggs was noted and bottles of brick, tile and paver cleaner and lime sulfur were found in the rear of the vehicle. Suicide notes were found in the house. At autopsy there was no evidence of significant trauma with black tarnishing of a silver-coloured ring and a silver-coloured necklace noted with staining of the underlying skin, in keeping with a chemical reaction between hydrogen sulfide and silver. Internally the most significant finding was unusual greenish discolouration of the gray matter of the external and cut surfaces of the cerebral hemispheres, cerebellum and brain stem. No other organs had this discoloration. Death was attributed to hydrogen sulfide poisoning. Skin discoloration from silver jewellery may represent a subtle external marker for lethal or non-lethal hydrogen sulfide exposure.

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Tree failure, or summer branch drop, refers to situations where large branches or whole trees may unexpectedly drop on unsuspecting victims. On occasion, this involves vehicles impacted by falling overhanging trees or branches. To study this phenomenon further review of the Forensic Science SA, Adelaide, Australia, database and an internet news search were undertaken over a 20 year period from March 2004 to February 2024 for all cases where unexpected accidental deaths in South Australia had been caused by falling branches or trees. There were six cases (age range 12-63 years; average 39 years; M:F 1:2). Three deaths occurred when branches dropped onto vehicles being driven along roads, 2 occurred when entire trees dropped onto the victims (one in windy weather) and 1 was due to a branch drop. Three tree species were identified as a grey box gum tree (Eucalyptus microcarpa), a lemon-scented gum tree (Corymbia citriodora) and a Cottonwood tree (Populus deltoides). The remainder were variants of eucalyptus (gum) trees. Deaths due to tree failures are uncommon and may involve a variety of different tree species with these events not necessarily occurring during high winds or storms. The major findings at autopsy are blunt force injuries.

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Hemibiotrophic fungi in the genus Colletotrichum employ a biotrophic phase invading host epidermal cells followed by a necrotrophic phase spreading through neighboring mesophyll and epidermal cells. We used serial block face scanning electron microscopy (SBF-SEM) to compare subcellular changes that occur in Medicago sativa (alfalfa) cotyledons during infection by Colletotrichum destructivum (compatible on M. sativa) and C. higginsianum (incompatible on M. sativa). Three-dimensional reconstruction of serial images revealed that alfalfa epidermal cells infected with C. destructivum undergo massive cytological changes during the first 60 hours following inoculation to accommodate extensive intracellular hyphal growth. Conversely, inoculation with the incompatible species C. higginsianum resulted in no successful penetration events and frequent formation of papilla-like structures and cytoplasmic aggregates beneath attempted fungal penetration sites. Further analysis of the incompatible interaction using focused ion beam-scanning electron microcopy (FIB-SEM) revealed formation of large multivesicular body-like structures that appeared spherical and were not visible in compatible interactions. These structures often fused with the host plasma membrane, giving rise to paramural bodies that appeared to be releasing extracellular vesicles (EVs). Isolation of EVs from the apoplastic space of alfalfa leaves at 60h post inoculation showed significantly more vesicles secreted from alfalfa infected with incompatible fungus compared to compatible fungus, which in turn was more than produced by non-infected plants. Thus, the increased frequency of paramural bodies during incompatible interactions correlated with an increase in EV quantity in apoplastic wash fluids. Together, these results suggest that EVs and paramural bodies contribute to immunity during pathogen attack in alfalfa.

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Chronic Traumatic Encephalopathy, or CTE, is an entity characterized by neurological deficits that are thought to arise from repetitive episodes of blunt head trauma. It has gained considerable attention recently in those who have engaged in contact sports. However, given that it is caused by mechanical cerebral strain from nonspecific blunt impact, it seems reasonable to assume that it could arise from a multitude of causes, such as craniocentric domestic violence. While the literature is somewhat contradictory, the possibilities are that CTE may be caused by either the incremental additive effects of less severe trauma, or from more forceful impacts, or from a combination of both of these mechanisms. Another issue to consider is the degree of acceleration/rotation trauma associated with particular events. Careful study of the chronology, nature and dose-relationships of previous head impacts in victims of inflicted lethal head trauma will, therefore, be required. This will help to clarify its significance in cases of domestic violence and also specifically whether it can be additive from more minor impacts, or whether there is a threshold of force required before it occurs.

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A 27-year-old male died suddenly due to cardiac tamponade arising from pericarditis complicating autoimmune polyglandular syndrome (APS) type 2. He had a history of primary Addison disease and autoimmune hypothyroidism which were corroborated at autopsy. In addition a florid fibrinous pericarditis was associated with 287 g of turbid fluid in the pericardial sac. Although pericarditis with tamponade is a potential complication of APS, it has rarely if ever, been reported as a cause of sudden death. Lethal mechanisms may involve both compressive and restrictive effects.

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Playing two-dimensional video games has been shown to result in improvements in a range of visual and cognitive tasks, and these improvements appear to generalize widely1,2,3,4,5,6. Here we report that young adults with healthy vision, surprisingly, showed a dramatic improvement in stereo vision after playing three-dimensional, but not two-dimensional, video games for a relatively short period of time. Intriguingly, neither group showed any significant improvement in binocular contrast sensitivity. This dissociation suggests that the visual enhancement was specific to genuine stereoscopic processing, not indirectly resulting from enhanced contrast processing, and required engaging in a disparity cue-rich three-dimensional environment.

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