Metallated phthalocyanines and their hydrophilic derivatives for multi-targeted oncological photodynamic therapy

Background and aim: A photosensitizer (PS) delivery and comprehensive tumor targeting platform was developed that is centered on the photosensitization of key pharmacological targets in solid tumors (cancer cells, tumor vascular endothelium, and cellular and non-cellular components of the tumor microenvironment) before photodynamic therapy (PDT). Interstitially targeted liposomes (ITLs) encapsulating zinc phthalocyanine (ZnPC) and aluminum phthalocyanine (AlPC) were formulated for passive targeting of the tumor microenvironment. In previous work it was established that the PEGylated ITLs were taken up by cultured cholangiocarcinoma cells. The aim of this study was to verify previous results in cancer cells and to determine whether the ITLs can also be used to photosensitize cells in the tumor microenvironment and vasculature. Following positive results, rudimentary in vitro and in vivo experiments were performed with ZnPC-ITLs and AlPC-ITLs as well as their water-soluble tetrasulfonated derivatives (ZnPCS4 and AlPCS4) to assemble a research dossier and bring this platform closer to clinical transition. Methods: Flow cytometry and confocal microscopy were employed to determine ITL uptake and PS distribution in cholangiocarcinoma (SK-ChA-1) cells, endothelial cells (HUVECs), fibroblasts (NIH-3T3), and macrophages (RAW 264.7). Uptake of ITLs by endothelial cells was verified under flow conditions in a flow chamber. Dark toxicity and PDT efficacy were determined by cell viability assays, while the mode of cell death and cell cycle arrest were assayed by flow cytometry. In vivo systemic toxicity was assessed in zebrafish and chicken embryos, whereas skin phototoxicity was determined in BALB/c nude mice. A PDT efficacy pilot was conducted in BALB/c nude mice bearing human triple-negative breast cancer (MDA-MB-231) xenografts. Results: The key findings were that (1) photodynamically active PSs (i.e., all except ZnPCS4) were able to effectively photosensitize cancer cells and non-cancerous cells; (2) following PDT, photodynamically active PSs were highly toxic-to-potent as per anti-cancer compound classification; (3) the photodynamically active PSs did not elicit notable systemic toxicity in zebrafish and chicken embryos; (4) ITL-delivered ZnPC and ZnPCS4 were associated with skin phototoxicity, while the aluminum-containing PSs did not exert detectable skin phototoxicity; and (5) ITL-delivered ZnPC and AlPC were equally effective in their tumor-killing capacity in human tumor breast cancer xenografts and superior to other non-phthalocyanine PSs when appraised on a per mole administered dose basis. Conclusions: AlPC(S4) are the safest and most effective PSs to integrate into the comprehensive tumor targeting and PS delivery platform. Pending further in vivo validation, these third-generation PSs may be used for multi-compartmental tumor photosensitization

Metallated phthalocyanines and their hydrophilic derivatives for multi-targeted oncological photodynamic therapy

Background and aim: A photosensitizer (PS) delivery and comprehensive tumor targeting platform was developed that is centered on the photosensitization of key pharmacological targets in solid tumors (cancer cells, tumor vascular endothelium, and cellular and non-cellular components of the tumor microenvironment) before photodynamic therapy (PDT). Interstitially targeted liposomes (ITLs) encapsulating zinc phthalocyanine (ZnPC) and aluminum phthalocyanine (AlPC) were formulated for passive targeting of the tumor microenvironment. In previous work it was established that the PEGylated ITLs were taken up by cultured cholangiocarcinoma cells. The aim of this study was to verify previous results in cancer cells and to determine whether the ITLs can also be used to photosensitize cells in the tumor microenvironment and vasculature. Following positive results, rudimentary in vitro and in vivo experiments were performed with ZnPC-ITLs and AlPC-ITLs as well as their water-soluble tetrasulfonated derivatives (ZnPCS4 and AlPCS4) to assemble a research dossier and bring this platform closer to clinical transition. Methods: Flow cytometry and confocal microscopy were employed to determine ITL uptake and PS distribution in cholangiocarcinoma (SK-ChA-1) cells, endothelial cells (HUVECs), fibroblasts (NIH-3T3), and macrophages (RAW 264.7). Uptake of ITLs by endothelial cells was verified under flow conditions in a flow chamber. Dark toxicity and PDT efficacy were determined by cell viability assays, while the mode of cell death and cell cycle arrest were assayed by flow cytometry. In vivo systemic toxicity was assessed in zebrafish and chicken embryos, whereas skin phototoxicity was determined in BALB/c nude mice. A PDT efficacy pilot was conducted in BALB/c nude mice bearing human triple-negative breast cancer (MDA-MB-231) xenografts. Results: The key findings were that (1) photodynamically active PSs (i.e., all except ZnPCS4) were able to effectively photosensitize cancer cells and non-cancerous cells; (2) following PDT, photodynamically active PSs were highly toxic-to-potent as per anti-cancer compound classification; (3) the photodynamically active PSs did not elicit notable systemic toxicity in zebrafish and chicken embryos; (4) ITL-delivered ZnPC and ZnPCS4 were associated with skin phototoxicity, while the aluminum-containing PSs did not exert detectable skin phototoxicity; and (5) ITL-delivered ZnPC and AlPC were equally effective in their tumor-killing capacity in human tumor breast cancer xenografts and superior to other non-phthalocyanine PSs when appraised on a per mole administered dose basis. Conclusions: AlPC(S4) are the safest and most effective PSs to integrate into the comprehensive tumor targeting and PS delivery platform. Pending further in vivo validation, these third-generation PSs may be used for multi-compartmental tumor photosensitization

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Long-term exposure to greenspace and metabolic syndrome: A Whitehall II study

Background Metabolic syndrome is an important risk factor for non-communicable diseases, particularly type 2 diabetes, coronary heart disease, and stroke. Long-term exposure to greenspace could be protective of metabolic syndrome, but evidence for such an association is lacking. Accordingly, we investigated the association between long-term exposure to greenspace and risk of metabolic syndrome. Methods The present longitudinal study was based on data from four clinical examinations between 1997 and 2013 in 6076 participants of the Whitehall II study, UK (aged 45–69 years at baseline). Long-term exposure to greenspace was assessed by satellite-based indices of greenspace including Normalized Difference Vegetation Index (NDVI) and Vegetation Continuous Field (VCF) averaged across buffers of 500 and 1000 m surrounding the participants’ residential location at each follow-up. The ascertainment of metabolic syndrome was based on the World Health Organization (WHO) definition. Hazard ratios for metabolic syndrome were estimated using Cox proportional hazards regression models, controlling for age, sex, ethnicity, lifestyle factors, and socioeconomic status. Results Higher residential surrounding greenspace was associated with lower risk of metabolic syndrome. An interquartile range increase in NDVI and VCF in the 500 m buffer was associated with 13% (95% confidence interval (CI): 1%, 23%) and 14% (95% CI: 5%, 22%) lower risk of metabolic syndrome, respectively. Greater exposure to greenspace was also associated with each individual component of metabolic syndrome, including a lower risk of high levels of fasting glucose, large waist circumference, high triglyceride levels, low HDL cholesterol, and hypertension. The association between residential surrounding greenspace and metabolic syndrome may have been mediated by physical activity and exposure to air pollution. Conclusions The findings of the present study suggest that middle-aged and older adults living in greener neighbourhoods are at lower risk of metabolic syndrome than those living in neighbourhoods with less greenspace

Residential surrounding greenness and cognitive decline: a 10-Year follow-up of the whitehall II cohort

BACKGROUND: Evidence on beneficial associations of green space with cognitive function in older adults is very scarce and mainly limited to cross-sectional studies. OBJECTIVES: We aimed to investigate the association between long-term residential surrounding greenness and cognitive decline. METHODS: This longitudinal study was based on three waves of data from the Whitehall II cohort, providing a 10-y follow-up (1997-1999 to 2007-2009) of 6,506 participants (45-68 y old) from the United Kingdom. Residential surrounding greenness was obtained across buffers of 500 and around the participants' residential addresses at each follow-up using satellite images on greenness (Normalized Difference Vegetation Index; NDVI) from a summer month in every follow-up period. Cognitive tests assessed reasoning, short-term memory, and verbal fluency. The cognitive scores were standardized and summarized in a global cognition z-score. To quantify the impact of greenness on repeated measurements of cognition, linear mixed effect models were developed that included an interaction between age and the indicator of greenness, and controlled for covariates including individual and neighborhood indicators of socioeconomic status (SES). RESULTS: In a fully adjusted model, an interquartile range (IQR) increase in NDVI was associated with a difference in the global cognition z-score of 0.020 [95% confidence interval (CI): 0.003, 0.037; p=0.02] in the 500-m buffer and of 0.021 (95% CI: 0.003, 0.039; p=0.02) in the 1,000-m buffer over 10 y. The associations with cognitive decline over the study period were stronger among women than among men. CONCLUSIONS: Higher residential surrounding greenness was associated with slower cognitive decline over a 10-y follow-up period in the Whitehall II cohort of civil servants

Green and blue spaces and physical functioning in older adults: Longitudinal analyses of the Whitehall II study

There is increasing evidence of the health benefits of exposure to natural environments, including green and blue spaces. The association with physical functioning and its decline at older age remains to be explored. The aim of the present study was to investigate the longitudinal association between the natural environment and the decline in physical functioning in older adults. We based our analyses on three follow-ups (2002-2013) of the Whitehall II study, including 5759 participants (aged 50 to 74 years at baseline) in the UK. Exposure to natural environments was assessed at each follow-up as (1) residential surrounding greenness across buffers of 500 and 1000 m around the participants' address using satellite-based indices of greenness (Enhanced Vegetation Index (EVI) and Normalized Difference Vegetation Index (NDVI)) and (2) the distance from home to the nearest natural environment, separately for green and blue spaces, using a land cover map. Physical functioning was characterized by walking speed, measured three times, and grip strength, measured twice. Linear mixed effects models were used to quantify the impact of green and blue space on physical functioning trajectories, controlled for relevant covariates. We found higher residential surrounding greenness (EVI and NDVI) to be associated with slower 10-year decline in walking speed. Furthermore, proximity to natural environments (green and blue spaces combined) was associated with slower decline in walking speed and grip strength. We observed stronger associations between distance to natural environments and decline in physical functioning in areas with higher compared to lower area-level deprivation. However, no association was observed with distance to green or blue spaces separately. The associations with decline in physical functioning were partially mediated by social functioning and mental health. Our results suggest that higher residential surrounding greenness and living closer to natural environments contribute to better physical functioning at older ages

Residential surrounding greenness and cognitive decline: a 10-Year follow-up of the whitehall II cohort

BACKGROUND: Evidence on beneficial associations of green space with cognitive function in older adults is very scarce and mainly limited to cross-sectional studies. OBJECTIVES: We aimed to investigate the association between long-term residential surrounding greenness and cognitive decline. METHODS: This longitudinal study was based on three waves of data from the Whitehall II cohort, providing a 10-y follow-up (1997-1999 to 2007-2009) of 6,506 participants (45-68 y old) from the United Kingdom. Residential surrounding greenness was obtained across buffers of 500 and around the participants' residential addresses at each follow-up using satellite images on greenness (Normalized Difference Vegetation Index; NDVI) from a summer month in every follow-up period. Cognitive tests assessed reasoning, short-term memory, and verbal fluency. The cognitive scores were standardized and summarized in a global cognition z-score. To quantify the impact of greenness on repeated measurements of cognition, linear mixed effect models were developed that included an interaction between age and the indicator of greenness, and controlled for covariates including individual and neighborhood indicators of socioeconomic status (SES). RESULTS: In a fully adjusted model, an interquartile range (IQR) increase in NDVI was associated with a difference in the global cognition z-score of 0.020 [95% confidence interval (CI): 0.003, 0.037; p=0.02] in the 500-m buffer and of 0.021 (95% CI: 0.003, 0.039; p=0.02) in the 1,000-m buffer over 10 y. The associations with cognitive decline over the study period were stronger among women than among men. CONCLUSIONS: Higher residential surrounding greenness was associated with slower cognitive decline over a 10-y follow-up period in the Whitehall II cohort of civil servants

Green and blue spaces and physical functioning in older adults: Longitudinal analyses of the Whitehall II study

There is increasing evidence of the health benefits of exposure to natural environments, including green and blue spaces. The association with physical functioning and its decline at older age remains to be explored. The aim of the present study was to investigate the longitudinal association between the natural environment and the decline in physical functioning in older adults. We based our analyses on three follow-ups (2002-2013) of the Whitehall II study, including 5759 participants (aged 50 to 74 years at baseline) in the UK. Exposure to natural environments was assessed at each follow-up as (1) residential surrounding greenness across buffers of 500 and 1000 m around the participants' address using satellite-based indices of greenness (Enhanced Vegetation Index (EVI) and Normalized Difference Vegetation Index (NDVI)) and (2) the distance from home to the nearest natural environment, separately for green and blue spaces, using a land cover map. Physical functioning was characterized by walking speed, measured three times, and grip strength, measured twice. Linear mixed effects models were used to quantify the impact of green and blue space on physical functioning trajectories, controlled for relevant covariates. We found higher residential surrounding greenness (EVI and NDVI) to be associated with slower 10-year decline in walking speed. Furthermore, proximity to natural environments (green and blue spaces combined) was associated with slower decline in walking speed and grip strength. We observed stronger associations between distance to natural environments and decline in physical functioning in areas with higher compared to lower area-level deprivation. However, no association was observed with distance to green or blue spaces separately. The associations with decline in physical functioning were partially mediated by social functioning and mental health. Our results suggest that higher residential surrounding greenness and living closer to natural environments contribute to better physical functioning at older ages

Thermodynamic profiling during irreversible electroporation in porcine liver and pancreas: a case study series

Aims: First, the aim of the study was to determine whether irreversible electroporation (IRE) is associated with heat generation in the liver and pancreas at clinical (≤1,500 V/cm) and supraclinical (>1,500 V/cm) electroporation settings; second, to assess the risk of thermal tissue damage in and adjacent to the treated volume in highly perfused versus moderately perfused parts of both organs; third, to investigate the influence of perfusion and of the presence and the orientation of a metal stent on the maximal thermal elevation (ΔTSession,max) in the tissue during an IRE session at fixed IRE settings, and finally, to determine whether the maximum temperature elevation within the IRE-subjected organ during an IRE treatment (single or multiple sessions) is reflected in the organ's surface temperature. Methods: The aims were investigated in 12 case studies conducted in five female Landrace pigs. Several IRE settings were applied for lateral (2), triangular (3), and rectangular (4) electrode configurations in the liver hilum, liver periphery, pancreas head, and pancreas tail. IRE series of 10-90 pulses were applied with pulse durations that varied from 70 μs to 90 μs and electric field strengths between 1,200 V/cm and 3,000 V/cm. In select cases, a metal stent was positioned in the bile duct at the level of the liver hilum. Temperatures were measured before, during, and after IRE in and adjacent to the treatment volumes using fiber optical temperature probes (temperature at the nucleation centers) and digital thermography (surface temperature). The occurrence of thermal damage was assumed to be at temperatures above 50 °C (ΔTSession,max ≥ 13 °C relative to body temperature of 37 °C). The temperature fluctuations at the organ surface (ΔTLocSurf) were compared to the maximum temperature elevation during an IRE treatment in the electroporation zone. In select cases, IRE was applied to tissue volumes encompassing the portal vein (PV) and a constricted and patent superior mesenteric vein (SMV) to determine the influence of the heatsink effect of PV and SMV on ΔTSession,max. Results: The median baseline temperature was 31.6 °C-36.3 °C. ΔTSession,max ranged from -1.7 °C to 25.5 °C in moderately perfused parts of the liver and pancreas, and from 0.0 °C to 5.8 °C in highly perfused parts. The median ΔTLocSurf of the liver and pancreas was 1.0 °C and 10.3 °C, respectively. Constricting the SMV in the pancreas head yielded a 0.8 °C higher ΔTSession,max. The presence of a metal stent in the liver hilum resulted in a ΔTSession,max of 19.8 °C. Stents parallel to the electrodes caused a ΔTSession,max difference of 4.2 °C relative to the perpendicular orientation. Conclusions: Depending on IRE settings and tissue type, IRE is capable of inducing considerable heating in the liver and pancreas that is sufficient to cause thermal tissue damage. More significant temperature elevations are positively correlated with increasing number of electrode pairs, electric field strength, and pulse number. Temperature elevations can be further exacerbated by the presence and orientation of metal stents. Temperature elevations at the nucleation centers are not always reflected in the organ's surface temperature. Heat sink effects caused by large vessels were minimal in some instances, possibly due to reduced blood flow caused by anesthesia. Relevance for patients: Appropriate IRE settings must be chosen based on the tissue type and the presence of stents to avoid thermal damage in healthy peritumoral tissue and to protect anatomical structures [Table: see text]