Hemorrhagic stroke is a debilitating neurological disease, affecting millions worldwide. Characterized by bleeding in the brain, it is caused by a breakdown of the blood-brain barrier (BBB) and causes damage through the presence of iron in the brain, immune activation and increased intracranial pressure. The goal of this mini-review is to explore the signaling pathways that lead to cell death that are a part of disease progression in hemorrhagic stroke. This mini-review will highlight clinical observations and data, while also incorporating findings using preclinical disease models. There are important roles for apoptosis, necroptosis, necrosis, autophagy, ferroptosis, and pyroptosis in hemorrhagic stroke. Recent work has highlighted the interplay between these phenomena, providing key regulators as potential therapeutic targets, including reactive oxygen species, iron metabolism, and caspases. Therapeutic strategies that can delay or counteract the cytotoxic effects of hemorrhage can improve clinical outcomes in hemorrhagic stroke patients.

BackgroundDuring the Covid-19 pandemic, people with chronic conditions experienced delayed or missed care, while their carers endured social isolation, loneliness, and reduced support. Information communication technology (ICT) can be utilized to encourage continuity of care, address misinformation, and allocate support. This study aimed to identify factors associated with the ICT adaptation of South Asian carers of individuals with chronic conditions by comparing changes in ICT utilization and preferences before and during the pandemic.Method416 South Asian carers reporting feelings of loneliness and isolation were identified from the Coping with Loneliness, Isolation and Covid-19 (CLIC) online survey. Descriptive statistics and multinomial regression models were utilized.ResultThe most commonly used ICT modality was auditory, followed by written and audio-visual. Four variables identified were: social network size and relationship proximity, Covid-19-induced distress, age, and living arrangements. We identified a negative correlation between social network size and ICT frequency/intensity, reductions in communication frequency/intensity associated with Covid-19-induced distress, working-age carer (18-60) preference adaptation toward written communication during the pandemic, written and auditory ICT fluency in carers spending time alone by choice, and aversion from auditory ICT in carers who lived and were often alone involuntarily.ConclusionThe findings provide insights into South Asian carers' ICT usage, preferences, and adaptation in response to the pandemic. The findings aid in the development of health and social care pathways that fulfil local caregivers' unmet support and resource needs.

In the mammalian isocortex, CD44, a cell surface receptor for extracellular matrix molecules, is present in pial-based and fibrous astrocytes of white matter but not in protoplasmic astrocytes. In the hominid isocortex, CD44+ astrocytes comprise the subpial "interlaminar" astrocytes, sending long processes into the cortex. The hippocampus also contains similar astrocytes. We have examined all levels of the human central nervous system and found CD44+ astrocytes in every region. Astrocytes in white matter and astrocytes that interact with large blood vessels but not with capillaries in gray matter are CD44+, the latter extending long processes into the parenchyma. Motor neurons in the brainstem and spinal cord, such as oculomotor, facial, hypoglossal, and in the anterior horn of the spinal cord, are surrounded by CD44+ processes, contrasting with neurons in the cortex, basal ganglia, and thalamus. We found CD44+ processes that intercalate between ependymal cells to reach the ventricle. We also found CD44+ astrocytes in the molecular layer of the cerebellar cortex. Protoplasmic astrocytes, which do not normally contain CD44, acquire it in pathologies like hypoxia and seizures. The pervasive and inducible expression of CD44 in astrocytes is a novel finding that lays the foundations for functional studies into the significance of CD44 in health and disease.

OBJECTIVE:Individuals with neurodevelopmental and/or neurocognitive disorders (NNDs) have complex, long-term care needs. In Bangladesh, India, and Pakistan, informal carers shoulder the responsibility and strain of providing care for people with NNDs. Intense care demand, societal and cultural care expectations, and lack of support infrastructure often lead to psychosocial strain in this inadequately researched community, particularly during crises such as the COVID-19 pandemic. This study explored and identified specific features of the coping styles exhibited by informal carers of people with NNDs from Bangladesh, India, and Pakistan during the COVID-19 pandemic. MATERIAL AND METHODS:Between June and November 2020, 245 carers in India, Pakistan, and Bangladesh responded to open-ended questions in the CLIC (Coping with Loneliness, Isolation, and COVID-19) survey. A reflexive thematic analysis was conducted to uncover the underlying themes and identify coping strategies and stressors. A frequency analysis was performed to examine the associations between these themes and carer nationality. Significant tests identified coping styles. RESULTS:We identified three coping styles: religiosity (Pakistan), caregiving as a natural life path (Bangladesh), and self-care (India). The religiosity and natural life path styles reside on the fatalism/acceptance continuum and suggest an insight-oriented therapeutic approach. Self-care is a problem-solving strategy that calls for a behaviorally oriented approach. Family overreliance on the carer was a concern across all three groups. CONCLUSIONS:The findings underscore the need for accessible support pathways to sustain care standards, ensuring the well-being of carers and care recipients.

The role of information asymmetry in automobile insurance markets has been studied extensively. However, not all empirical studies found correlation between driver risk and chosen coverage. There are two explanations for this (Cohen and Siegelman 2010 ): (1) information asymmetry may not characterise environments in which consumers cannot act on private information; (2) most studies measure not risky driver behaviour, but ex post realisations of such behaviour, e.g. claims. In Taiwan’s taxi insurance market, insurers cannot freely use rating factors. Usage density (e.g. daily distance), a direct measure of risk, therefore constitutes private information. Using ordinary least squares models, we find that usage density is positively correlated with total premiums. Using logistic models, we find high-density drivers more likely to buy additional policies. Thus, usage density is positively correlated with insurance demand, consistent with the risk-coverage hypothesis. Unlimited-mileage coverage may disproportionately attract high-risk drivers, an argument in favour of usage-based insurance.

PurposeRetinal vein occlusion (RVO) is a sight-threatening condition typically treated with intravitreal injection of vascular endothelial growth factor (VEGF) antagonists. Treatment response to anti-VEGF therapies is highly variable, with poor visual outcomes and treatment response in patients with significant retinal nonperfusion following RVO. Recently, caspase-9 has been identified as a potent regulator of edema, gliosis, and neuronal dysfunction during acute retinal hypoxia. The purpose of this study was to compare the therapeutic effect of caspase-9 inhibition against VEGF-neutralization in an established mouse model of RVO. MethodsAdult male C57Bl/6 J mice were randomized to induction of RVO and treatment with either vehicle, intravitreal injection of anti-VEGF antibody, topical administration of a selective caspase-9 inhibitor (Pen1-XBir3), or a combination therapy. Animals were followed on days 1, 2, and 8 after RVO with fundus retinal imaging, and with optical coherence tomography (OCT) to capture retinal swelling, capillary nonperfusion (measured by disorganization of retinal inner layers, DRIL), hyperreflective foci (HRF), and retinal atrophy. Focal electroretinography (ERG) measurements were performed on day 7. Histology was performed on retinal sections from day 8. ResultsBoth VEGF neutralization and caspase-9 inhibition showed significant retinal protection from RVO compared to vehicle treatment arm. Retinal reperfusion of occluded veins was accelerated in eyes receiving caspase-9 inhibitor, but not significantly different from vehicle in the anti-VEGF group. Retinal edema was suppressed in all treatment groups, with approximately 2-fold greater edema reduction with caspase-9 inhibition compared to VEGF neutralization. HRF were reduced similarly across all treatment groups compared to vehicle. Retinal detachment was reduced only in eyes treated with caspase-9 inhibitor monotherapy. Caspase-9 inhibition reduced retinal atrophy and preserved ERG response; VEGF neutralization did not prevent neurodegeneration following RVO. ConclusionCaspase-9 inhibition confers stronger neuronal and vascular protection compared to VEGF neutralization in the mouse laser-induced model of RVO.

The enzyme caspase-2 plays a central role in cellular apoptosis and is involved in the early stages of β-amyloid 42 and tau deposition, making it a promising therapeutic target for Alzheimer’s Disease (AD). Caspase-2 is highly expressed in the brains of patients with AD ( 1, 4 ) and those with mild cognitive impairment, which often precedes AD. This indicates the upregulation of caspase-2 early in disease progression, which is not observed in healthy adult brains ( 1-3 ). New irreversible small peptidomimetics (LJ2a and LJ3a) inhibit human caspase-2 with a remarkably high inactivation rate. ( 5 ) In primary hippocampal neurons treated with β-amyloid oligomers, submicromolar concentrations of LJ2a and LJ3a prevent synapse loss. Using a tamoxifen-inducible Cre system for the global removal of caspase-2, a series of experiments were performed on mice stereotaxically injected with β-amyloid 42. This model was used to validate caspase-2 blockade by assessing the cellular specificity of caspase-2 activation, the induction of downstream targets, and cell death marker expression via immunohistochemistry and western blot. Replication of these experiments remains underway. Presentation of current data can inform the rationality for using caspase-2 inhibitors as a therapeutic treatment. References : 1. Jean, Y. Y. et al. Caspase-2 is essential for c-Jun transcriptional activation and Bim induction in neuron death. Biochem J 455, 15-25, https://doi.org/10.1042/BJ20130556 (2013). 2. Wang, L., Miura, M., Bergeron, L., Zhu, H. & Yuan, J. Ich-1, an Ice/ced-3-related gene, encodes both positive and negative regulators of programmed cell death. Cell 78, 739-750 (1994). 3. Kumar, S., Kinoshita, M., Noda, M., Copeland, N. G. & Jenkins, N. A. Induction of apoptosis by the mouse Nedd2 gene, which encodes a protein similar to the product of the Caenorhabditis elegans cell death gene ced-3 and the mammalian IL-1 beta-converting enzyme. Genes & development 8, 1613-1626 (1994). 4. Liu, P. et al. A soluble truncated tau species related to cognitive dysfunction is elevated in the brain of cognitively impaired human individuals. Sci Rep 10, 3869, https://doi.org/10.1038/s41598-020-60777-x (2020). 5. Bosc, E., Anastasie, J., Soualmia, F. et al. Genuine selective caspase-2 inhibition with new irreversible small peptidomimetics. Cell Death Dis 13, 959. https://doi.org/10.1038/s41419-022-05396-2 (2022).

Apoptosis is a form of regulated cell death (RCD) that involves proteases of the caspase family. Pharmacological and genetic strategies that experimentally inhibit or delay apoptosis in mammalian systems have elucidated the key contribution of this process not only to (post-)embryonic development and adult tissue homeostasis, but also to the etiology of multiple human disorders. Consistent with this notion, while defects in the molecular machinery for apoptotic cell death impair organismal development and promote oncogenesis, the unwarranted activation of apoptosis promotes cell loss and tissue damage in the context of various neurological, cardiovascular, renal, hepatic, infectious, neoplastic and inflammatory conditions. Here, the Nomenclature Committee on Cell Death (NCCD) gathered to critically summarize an abundant pre-clinical literature mechanistically linking the core apoptotic apparatus to organismal homeostasis in the context of disease.

Caspase-2 (Casp2) is a promising therapeutic target in several human diseases, including nonalcoholic steatohepatitis (NASH) and Alzheimer’s disease (AD). However, the design of an active-site-directed inhibitor selective to individual caspase family members is challenging because caspases have extremely similar active sites. Here we present new peptidomimetics derived from the VDVAD pentapeptide structure, harboring non-natural modifications at the P2 position and an irreversible warhead. Enzyme kinetics show that these new compounds, such as LJ2 or its specific isomers LJ2a, and LJ3a, strongly and irreversibly inhibit Casp2 with genuine selectivity. In agreement with the established role of Casp2 in cellular stress responses, LJ2 inhibits cell death induced by microtubule destabilization or hydroxamic acid-based deacetylase inhibition. The most potent peptidomimetic, LJ2a, inhibits human Casp2 with a remarkably high inactivation rate ( k 3 / K i ~5,500,000 M −1 s − 1 ), and the most selective inhibitor, LJ3a, has close to a 1000 times higher inactivation rate on Casp2 as compared to Casp3. Structural analysis of LJ3a shows that the spatial configuration of C α at the P2 position determines inhibitor efficacy. In transfected human cell lines overexpressing site-1 protease (S1P), sterol regulatory element-binding protein 2 (SREBP2) and Casp2, LJ2a and LJ3a fully inhibit Casp2-mediated S1P cleavage and thus SREBP2 activation, suggesting a potential to prevent NASH development. Furthermore, in primary hippocampal neurons treated with β-amyloid oligomers, submicromolar concentrations of LJ2a and of LJ3a prevent synapse loss, indicating a potential for further investigations in AD treatment.

Advancements in ophthalmic imaging tools offer an unprecedented level of access to researchers working with animal models of neurovascular injury. To properly leverage this greater translatability, there is a need to devise reproducible methods of drawing quantitative data from these images. Optical coherence tomography (OCT) imaging can resolve retinal histology at micrometer resolution and reveal functional differences in vascular blood flow. Here, we delineate noninvasive vascular readouts that we use to characterize pathological damage post vascular insult in an optimized mouse model of retinal vein occlusion (RVO). These readouts include live imaging analysis of retinal morphology, disorganization of retinal inner layers (DRIL) measure of capillary ischemia, and fluorescein angiography measures of retinal edema and vascular density. These techniques correspond directly to those used to examine patients with retinal disease in the clinic. Standardizing these methods enables direct and reproducible comparison of animal models with clinical phenotypes of ophthalmic disease, increasing the translational power of vascular injury models.

The family of caspases is known to mediate many cellular pathways beyond cell death, including cell differentiation, axonal pathfinding, and proliferation. Since the identification of the family of cell death proteases, there has been a search for tools to identify and expand the function of specific family members in development, health, and disease states. However, many of the currently commercially available caspase tools that are widely used are not specific for the targeted caspase. In this report, we delineate the approach we have used to identify, validate, and target caspase-9 in the nervous system using a novel inhibitor and genetic approaches with immunohistochemical read-outs. Specifically, we used the retinal neuronal tissue as a model to identify and validate the presence and function of caspases. This approach enables the interrogation of cell-type specific apoptotic and non-apoptotic caspase-9 functions and can be applied to other complex tissues and caspases of interest. Understanding the functions of caspases can help to expand current knowledge in cell biology, and can also be advantageous to identify potential therapeutic targets due to their involvement in disease.

ABSTRACT Retinal glial cells— microglia, astrocytes, and Müller glia—provide homeostatic support, regulate vascular blood flow, and react to injury by releasing inflammatory cytokines. Glial reactivity has been shown to be relevant for retinal vascular pathology and neuronal death. Non-apoptotic expression of endothelial caspase-9 (EC Casp9) was recently identified as a key mediator of retinal edema, hypoxic-ischemic injury, and neurodegeneration in retinal vein occlusion (RVO). In the current study we aimed to determine the glial responses that are modulated by EC Casp9 as a means to identify relevant neuro-immune mechanisms for the development of retinal edema and neurodegeneration. To this end we used a mouse model of RVO and a tamoxifen inducible EC Casp9 KO mouse line. We show that EC Casp9 leads to an increase in reactive microglia and to macrogliosis in a time-dependent manner. RVO induced an EC Casp9 dependent astroglial caspase-6 and cleavage of GFAP. Cytokine array analysis revealed that RVO increases expression of inflammatory cytokines out of which CX3CL1, IGF-1, IL-4, LIX, IL-1α, M-CSF, TNF-α, IL-1β, IL-10, and VEGF-A, were regulated by EC Casp9. Moreover, we found that EC Casp9 deletion resulted in protection from contrast sensitivity decline one day post-RVO. These results demonstrate that caspase-9 in hypoxic endothelial cells regulates retinal inflammatory signaling in microglia, astrocytes and Müller cells and changes in visual function.

Purpose: To characterize postnatal ocular pathology in a Ndufs4(-/-) mouse model of complex I deficiency using noninvasive retinal imaging and visual testing. Methods: Ndufs4(-/-) mice and wild-type (WT) littermates were analyzed at 3, 5, and 7 weeks postnatal. Retinal morphology was visualized by optical coherence tomography (OCT). OCT images were analyzed for changes in retinal thickness and reflectivity profiles. Visual function was assessed by electroretinogram (ERG) and optomotor reflex (OMR). Results: Ndufs4(-/-) animals have normal OCT morphology at weaning and develop innerplexiformlayer atrophy overweeks 5 to 7. Outer retinal layers showhyporeflectivity of the external limiting membrane (ELM) and photoreceptor ellipsoid zone (EZ). Retinal function is impaired at 3weeks, with profound deficits in b-wave, a-wave, and oscillatory potential amplitudes. The b-wave and oscillatory potential implicit times are delayed, but the a-wave implicit time is unaffected. Ndufs4(-/-) animals have normal OMR at 3 weeks and present with increasing acuity and contrast OMR deficits at 5 and 7 weeks. Physiological thinning of inner retinal layers, attenuation of ELM reflectivity, and attenuation of ERG b- and a-wave amplitudes occur inWT C57BL/6 littermates between weeks 3 and 7. Conclusions: Noninvasive ocular imaging captures early-onset retinal degeneration in Ndufs4(-/-) mice and is a tractable approach for investigating retinal pathology subsequent to complex I deficiency. Translational Relevance: Ophthalmic imaging captures clinically relevant measures of retinal disease in a fast-progressing mouse model of complex I deficiency consistent with human Leigh syndrome.

The COVID-19 pandemic has renewed attention to the conflicting demands placed on caregivers worldwide. Nowhere is this more evident than in South Asia, where cultural norms and a scarcity of long-term care facilities have made the home the locus of caregiving for individuals with brain health conditions and disabilities. A hidden cost of this informal arrangement is the burden experienced by the primary caregiver. This may be exacerbated when traditional caregiving expectations clash with personal aspirations outside the family. This paper investigates the determinants of burden among 494 informal caregivers in India, Pakistan, and Bangladesh during the COVID-19 pandemic. The variables, obtained from the Coping with Loneliness, Isolation, and COVID-19 (CLIC) Global Caregiver study, include caregiver demographic characteristics, features of the caregiving arrangement, and subjective measures of the caregiving experience. The dependent variable in our analysis, the perceived frequency of burden, is rated by respondents on a five-level scale. Role strain is expected to be greatest among young, educated women living with the care recipient in their own homes. Therefore, burden should exhibit a positive association with female gender, education, and cohabitation, and a negative association with age. Out of the 494 caregivers included in the study, 33.8% experienced increasing burden and 58% experienced loneliness some of the time during Covid-19. The ordinal logistic marginal effects are consistent with all four hypotheses. Caregiver Burden is found to be positively correlated with female gender (OR,1.92; 95%CI, 0.35,1,03; P =<0.001), the 18-39 age range (OR,1.71; 95%CI, 0.114, 0,963; P =0.01), poor finances (OR,2.192; 95%CI,0.22,1.34; P =0.006), poor physical health (OR, 2.24, 95%CI, 0.29,1.32, P =0.002), poor mental health (OR,1.86; 95%CI, 0.14,1.10, P =0.011), and care recipients with dementia/Alzheimer's (OR,2.76; 95%CI, 0.52,1.51, P =<0.001) and/or mental health conditions (OR,1.61; 95%CI, 0.02,0.94, P =0.039). Role strain may be a significant driver of burden when the caretaker’s educational and professional potential collide with traditional South Asian family obligations. The burden reported by our respondents suggests that role strain may elevate stress among young, educated caregivers. Our results, therefore, provide indirect evidence concerning the changing economic and socio-cultural context of caregiving in South Asian households.

In albinism, aberrations in the ipsi-/contralateral retinal ganglion cell (RGC) ratio compromise the functional integrity of the binocular circuit. Here, we focus on the mouse ciliary margin zone (CMZ), a neurogenic niche at the embryonic peripheral retina, to investigate developmental processes regulating RGC neurogenesis and identity acquisition. We found that the mouse ventral CMZ generates predominantly ipsilaterally projecting RGCs, but this output is altered in the albino visual system because of CyclinD2 downregulation and disturbed timing of the cell cycle. Consequently, albino as well as CyclinD2-deficient pigmented mice exhibit diminished ipsilateral retinogeniculate projection and poor depth perception. In albino mice, pharmacological stimulation of calcium channels, known to upregulate CyclinD2 in other cell types, augmented CyclinD2-dependent neurogenesis of ipsilateral RGCs and improved stereopsis. Together, these results implicate CMZ neurogenesis and its regulators as critical for the formation and function of the mammalian binocular circuit.

Introduction: Families increasingly employ foreign domestic workers (FDWs) to care for older loved ones. Caregiver burden reflects FDWs' difficulty adapting to work demands. We test hypothesized associations between burden and six personal characteristics: children, marriage, education, Chinese proficiency, eldercare experience, and non-eldercare experience. Method: In total, 299 Indonesian FDWs in Taiwan completed the Zarit Burden Interview. Exploratory factor analysis identified the dimensions of burden. Multiple and multinomial regressions related the variables to overall burden, burden dimensions, and burden severity. Results: Four dimensions were found: personal strain, role strain, dependency, and guilt. Children were negatively associated with burden, role strain, dependency, and guilt. Chinese proficiency was negatively associated with severity and guilt. Eldercare experience was positively associated with severity and personal strain. Marriage was non-monotonically related to severity. Conclusion: Caregivers whose earnings benefit their children may be more likely to thrive in Taiwan. Language training may boost caregiver performance and host family satisfaction.

Retinal neurovascular injuries are a leading cause of vision loss in young adults presenting unmet therapeutic needs. Neurovascular injuries damage homeostatic communication between endothelial, pericyte, glial, and neuronal cells through signaling pathways that remain to be established. To understand the mechanisms that contribute to neuronal death, we use a mouse model of retinal vein occlusion (RVO). Using this model, we previously discovered that after vascular damage, there was non-apoptotic activation of endothelial caspase-9 (EC Casp9); knock-out of EC Casp9 led to a decrease in retinal edema, capillary ischemia, and neuronal death. In this study, we aimed to explore the role of EC Casp9 in vision loss and inflammation. We found that EC Casp9 is implicated in contrast sensitivity decline, induction of inflammatory cytokines, and glial reactivity. One of the noted glial changes was increased levels of astroglial cl-caspase-6, which we found to be activated cell intrinsically by astroglial caspase-9 (Astro Casp9). Lastly, we discovered that Astro Casp9 contributes to capillary ischemia and contrast sensitivity decline after RVO (P-RVO). These findings reveal specific endothelial and astroglial non-apoptotic caspase-9 roles in inflammation and neurovascular injury respectively; and concomitant relevancy to contrast sensitivity decline.

Mouse models of retinal vein occlusion (RVO) are often used in ophthalmology to study hypoxic-ischemic injury in the neural retina. In this report, a detailed method pointing out critical steps is provided with recommendations for optimization to achieve consistently successful occlusion rates across different genetically modified mouse strains. The RVO mouse model consists primarily of the intravenous administration of a photosensitizer dye followed by laser photocoagulation using a retinal imaging microscope attached to an ophthalmic guided laser. Three variables were identified as determinants of occlusion consistency. By adjusting the wait time after rose bengal administration and balancing the baseline and experimental laser output, the variability across experiments can be limited and a higher success rate of occlusions achieved. This method can be used to study retinal diseases that are characterized by retinal edema and hypoxic-ischemic injury. Additionally, as this model induces vascular injury, it can also be applied to study the neurovasculature, neuronal death, and inflammation.