Damage To Vasculature Research Articles

Gender-related differences in the timing of intervention for aortic stenosis: focus on cardiac damage

Abstract Background and aim: There is an increasing awareness of gender-related differences in the management of cardiovascular diseases. Our aim was to assess whether cardiac damage prior to intervention for severe aortic stenosis (AS) differs according to gender. Methods Retrospective analysis of a prospective cohort of patients with AS who underwent surgical aortic valve replacement (SAVR) and transcatheter aortic valve replacement (TAVR) in a tertiary care center from 2017 to 2022. Echocardiographic parameters prior to surgery were collected, and patients were classified into categories of a cardiac damage staging system. (1) Several echocardiographic variables according to gender were evaluated. Results 869 patients were included, of which 350 underwent TAVR and 519 were treated with SAVR. Females represented 52.4% (n=272) of TAVR patients and 37.7% (n=132) of SAVR patients. In the echocardiogram performed before the intervention, both in the TAVR and the SAVR cohort, female patients had worse diastolic function (significantly higher values of E/e’), and greater involvement of pulmonary vasculature damage (significantly higher pulmonary artery systolic pressure and worse right ventricular-arterial coupling (Table-Figure 1). When applying an available cardiac damage staging system (1), we documented that women were treated at a statistically significant more advanced stage compared to men (Figure 2). Conclusions There are gender-related differences in the timing of aortic stenosis intervention, both with TAVR and SAVR. Women are operated on at a more advanced stage of cardiac damage, particularly regarding the parameters of diastolic function and right-side damage. These findings should be taken into account to improve intervention timing and outcomes, especially for women.Table-Figure 1Figure 2

7770 Damage of Ovarian Vasculature by Chemotherapies induces Ovarian Dysfunction in Mice

Abstract Disclosure: W. So: None. R. Dong: None. Y. Luan: None. A. Abazarikia: None. S. Kim: None. Background: Chemotherapy demonstrates efficacy to destroying cancer cells; however, it can inadvertently impact normal healthy cells, leading to irreversible side effects, particularly affecting the ovaries in female patients. Female cancer survivors encounter endocrine dysfunction and infertility due to ovarian damage caused by gonadotoxic cancer therapies. The ovary, a vital organ responsible for producing oocytes and endocrine hormones, faces significant challenges following such treatment. A study with premenopausal breast cancer patients reported a significant reduction in ovarian blood flow and volume immediately following chemotherapy. The preservation of ovarian function depends on the ovarian vasculature in supporting the development and maintenance of ovarian follicles. Despite its importance, the relationship between ovarian vasculature and follicle following chemotherapy has not been studied. Results: To investigate the direct impacts of chemotherapy, ovaries from postnatal 5 CD-1 mice were subjected to testing with three chemotherapeutic agents: cisplatin (CDDP), cyclophosphamide (CPA), and doxorubicin (DOXO). The expression of CD31/PECAM-1 (platelet endothelial adhesion molecule) exhibited a significant decrase after 96 hours of culture with 4-HC, an active metabolite of CPA. This reduction implies that ovarian vasculature was also compromised when chemotherapy eliminated immature follicles. In order to evaluate the influence of vascular endothelial growth factor (VEGF) in safeguarding both vasculature and ovarian follicles, ovaries were co-cultured with VEGF165 or VEGF165b in combination with chemotherapy. Interestingly, pre-treatment with VEGF165 before 4-HC led to an increased expression of CD31 in the cortex area, while VEGF165b further diminished CD31 expression compared to treatment with 4-HC alone. Additionally, adolescent CD-1 mice were intraperitoneally injected with CDDP, CPA, or DOXO, and ovaries were harvested one week after injection. Ovaries treated with chemotherapeutic agents exhibited a reduction in ovary weight and decreased CD31 expression compared to control mice. Conclusions: We propose that these chemotherapeutic agents not only induced the loss of ovarian follicles but also affected ovarian vasculature in mice. This suggests a potential decrease in folliculogenesis within the ovary, coupled with the depletion of ovarian reserve, as the delivery of nutrients and oxygen to growing ovarian follicles may be compromised. VEGF165 may play a beneficial role in maintaining ovarian function by safeguarding ovarian vasculature following chemotherapy. Understanding these intricate dynamics is essential for developing strategies to mitigate the negative impact of cancer therapies on reproductive health in female cancer survivors, particularly in preserving endocrine function and reproductive lifespan. Presentation: 6/2/2024

Protective Effect of DHQ-11 against Hypoxia-induced Vasorelaxation

Hypoxia, or the lack of oxygen, has multiple impacts on the vascular system. The major molecular sensors for hypoxia at the cellular level are hypoxia inducible factor and heme oxygenase. Hypoxia also acts on the vasculature directly conveying its damaging effects through disruption of the control of vascular tone, particularly in the coronary circulation, enhancement of inflammatory responses and activation of coagulation pathways. These effects could be particularly detrimental under pathological conditions such as obstructive sleep apnea and other breathing disorders. Introduction: The effect of conjugate 2-(3,4-Dihydroxyphenyl)-6-{[1-(2’-bromo-3’,4’-dimethoxyphenyl)-6,7-dimethoxy-3,4-dihydroisoquinolin-2(1H )-yl]methyl}-3,5,7-trihydroxychroman-4-one (DHQ-11) on hypoxia-induced vasorelaxation was investigated in rat aortic rings using standard organ bath techniques. Materials and methods: Hypoxia was stimulated by a superfusion of aortic rings with a glucose-free Krebs solution bubbled with 95 % N2/5 % CO2. The effect of conjugate DHQ-11 was assessed after a 60- min period of hypoxia on aortic rings precontracted with 50 mm KCl or 1µM phenylephrine (PE). The conjugate DHQ-11 significantly attenuated hypoxia-induced vasorelaxation in the endothelium-intact aortic rings precontracted with KCl or PE in a concentration-dependent manner. Results and discussion: This effect of conjugate DHQ-11 was more potent in aortic rings precontracted with PE than those with KCl where it maximally reduced hypoxia-induced vasorelaxation from 44.7 ± 3.7 to 5.4 ± 3.7 and 33.9 ± 3.4 to 10.8 ± 4.2 %, respectively. The removal of the endothelium attenuated the effect of conjugate DHQ-11 on hypoxia-induced vasorelaxation. Similarly, pretreatment of endothelium-intact aortic rings with L-NAME and methylene blue also attenuated the effect of conjugate DHQ-11 on hypoxia-induced vasorelaxation. Furthermore, the blockade of the ATP-sensitive KATP channel with glibenclamide and the calcium-activated large conductance BKCa channel with TEA also significantly attenuated the effect of conjugate DHQ-11 on hypoxia-induced vasorelaxation. Collectively, these results indicated that conjugate DHQ-11 attenuated the hypoxia-induced vasorelaxation suggesting that it alleviated the oxidative damage of vasculature. Conclusions: This effect of conjugate DHQ-11 possible is mediated through several mechanisms including the blockage of the extracellular Ca2+ entry via the voltage-dependent and receptor-operative Ca2+ channels, as well as inhibition of sarcoplasmic reticulum Ca2+ release via the inositol triphosphate pathway. In addition, endothelium and NO/sGC/cGMP/PKG pathway, as well as KATP and BKCa channels most likely participate in protection by conjugate DHQ-11 against hypoxia-induced vasorelaxation.

Deciphering the connection between microvascular damage and neurodegeneration in early diabetic retinopathy.

Diabetic retinopathy (DR), a common diabetes complication leading to vision loss, presents early clinical signs linked to retinal vasculature damage, affecting the neural retina at advanced stages. However, vascular changes and potential effects on neural cells before clinical diagnosis of DR are less well understood. To study the earliest stages of DR we performed histological phenotyping and quantitative analysis on postmortem retinas from 10 donors with diabetes and without signs of DR (such as microaneurysms and haemorrhages), plus 3 controls and 1 DR case, focusing on capillary loss in the deeper (DVP) and superficial vascular plexuses (SVP) and neural retina effects. The advanced DR case exhibited profound vascular and neural damage, whereas the ten randomly selected donors with diabetes appeared superficially normal. The SVP was indistinguishable from the controls. In contrast, over half of the retinas from donors with diabetes showed capillary dropout in the DVP and increased capillary diameter. However, we could not detect any localised neural cell loss in the vicinity of dropout capillaries. Instead, we observed a subtle pan-retinal loss of inner nuclear layer (INL) cells in all diabetes cases (p<0.05), independent of microvascular damage. In conclusion, our findings demonstrate a novel histological biomarker for early-stage diabetes-related damage in human postmortem retina, common in people with diabetes before clinical DR diagnosis. Furthermore, the mismatch between capillary dropout and neural loss questions the notion of microvascular loss directly causing neurodegeneration at the earliest stages of DR, so diabetes may affect the two readouts independently.

Neutrophil-inflicted vasculature damage suppresses immune-mediated optic nerve regeneration

In adult mammals, injured retinal ganglion cells (RGCs) fail to spontaneously regrow severed axons, resulting in permanent visual deficits. Robust axon growth, however, is observed after intra-ocular injection of particulate β-glucan isolated from yeast. Blood-borne myeloid cells rapidly respond to β-glucan, releasing numerous pro-regenerative factors. Unfortunately, the pro-regenerative effects are undermined by retinal damage inflicted by an overactive immune system. Here, we demonstrate that protection of the inflamed vasculature promotes immune-mediated RGC regeneration. In the absence of microglia, leakiness of the blood-retina barrier increases, pro-inflammatory neutrophils are elevated, and RGC regeneration is reduced. Functional ablation of the complement receptor 3 (CD11b/integrin-αM), but not the complement components C1q-/- or C3-/-, reduces ocular inflammation, protects the blood-retina barrier, and enhances RGC regeneration. Selective targeting of neutrophils with anti-Ly6G does not increase axogenic neutrophils but protects the blood-retina barrier and enhances RGC regeneration. Together, these findings reveal that protection of the inflamed vasculature promotes neuronal regeneration.

Abstract C116: Impact of human iPSC-derived MSC secretome in modulating radiation induced inflammation and brain vascular endothelial cell damage

Abstract Introduction: Radiation therapy (RT) is the standard of care for central nervous system (CNS) cancers. Despite the success of RT in reducing tumor mass, RT-induced side effects such as ECM remodeling, vasculopathies and neuroinflammation are well documented that cause reduced neurogenecity and premature aging like complications during post-cancer survivorship. Since mesenchymal stromal cells (MSCs) are multipotent adult stem cells that support tissue integrity, homeostasis and repair through factors they release, we here investigated the potential of iPSC-derived MSC (iMSC)-secretome in immunomodulation and vasculature repair in response to radiation-induced alterations utilizing models of human monocytic and microglial cells (THP1, HMC-3) and human brain endothelial cells (HCMEC/D3). Approaches: (a) Investigate the impact of iMSC secretome on vasculature damage caused by radiation-induced cell injury and RT-induced inflammation, by evaluating its effects on (i) endothelial cell survival and morphogenesis (ii) activation of peripheral monocytes stimulated by X-ray radiation therapy (RT), RT-induced microglial injury, and vasculature injury mediated inflammation (b) Identify factors and pathways that may contribute to the observed phenotype. Results: Decline in intact angiogenic tube formation and less compact spheroid formation was observed for HCMEC/D3 cells after irradiation at doses (Gy): 2.5, 5, 10, 15, and 30, with maximal effect observed at 30Gy. Evaluating the effects of iMSC secretome on radiation induced endothelial cell damage and inflammation revealed increased cell viability with reduced apoptosis, higher cell adherance and better spheroid compactness in 5Gy irradiated HCMEC/D3 cells after iMSC-conditioned media (iMSC-CM) treatment. Reduction in NFκB promoter activity was observed in irradiated THP1- NFκB-Luc2 reporter line, with reduced levels of TNF-α release per unit cell viability in THP1 after iMSC-CM treatment. NFκB promoter activity was also reduced in effect of iMSC treatment in THP1- NFκB-Luc2 reporter cells subjected to irradiated vasculature cells (D3), with or without iMSC coculture and, irradiated microglial cells (HMC3), mimicking vasculature injury, microglial injury, and immune cell crosstalk, respectively. iMSC secretome facilitated a more networked tube formation in HCMEC/D3 cells in coculture of D3+iMSC cells that was longer retained. Cytokine arrays revealed iMSC-CM to have proangiogenic, immunomodulatory (immune suppressive) and chemotactic factors: MCP1, ANG (angiogenin), IL6, IL8, GRO, GROα, RANTES, and SDF1. Our initial pathway analysis using human phospho-proteome profiling showed iMSC secretome elevates levels of beta-catenin, and PRAS40 (AKT/mTOR modulator) and lowers level of pRSK1/2/3 in HCMEC/D3. Conclusions: Taken together, we demonstrated that iMSC-secretome complements the proangiogenic and immunosuppressive effects of iMSC during radiation induced injury. Further studies are required to validate the in vivo potential of iMSC-secretome as safe biological therapy in long-term cancer survivors. Citation Format: Kshama Gupta, Ralph B. Perkerson III, Tammee M Parsons, Prasanna Vibhute, Abba C. Zubair, Alfredo Quinones-Hinojosa, Takahisa Kanekiyo. Impact of human iPSC-derived MSC secretome in modulating radiation induced inflammation and brain vascular endothelial cell damage [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr C116.

Loss of glyoxalase 2 alters the glucose metabolism in zebrafish

Glyoxalase 2 is the second enzyme of the glyoxalase system, catalyzing the detoxification of methylglyoxal to d-lactate via SD-Lactoylglutathione. Recent in vitro studies have suggested Glo2 as a regulator of glycolysis, but if Glo2 regulates glucose homeostasis and related organ specific functions in vivo has not yet been evaluated. Therefore, a CRISPR-Cas9 knockout of glo2 in zebrafish was created and analyzed. Consistent with its function in methylglyoxal detoxification, SD-Lactoylglutathione, but not methylglyoxal accumulated in glo2−/− larvae, without altering the glutathione metabolism or affecting longevity. Adult glo2−/− livers displayed a reduced hexose concentration and a reduced postprandial P70–S6 kinase activation, but upstream postprandial AKT phosphorylation remained unchanged. In contrast, glo2−/− skeletal muscle remained metabolically intact, possibly compensating for the dysfunctional liver through increased glucose uptake and glycolytic activity. glo2−/− zebrafish maintained euglycemia and showed no damage of the retinal vasculature, kidney, liver and skeletal muscle. In conclusion, the data identified Glo2 as a regulator of cellular energy metabolism in liver and skeletal muscle, but the redox state and reactive metabolite accumulation were not affected by the loss of Glo2.

Preservation of vasculature is beneficial in mitigating cisplatin induced acute kidney injury

Abstract Introduction Cisplatin, discovered over 50 years ago, is still one of the most potent anti-cancer chemotherapy drugs and broadly used in first-line chemotherapy regimens subscribed to about 10 to 20% of total cancer patients. However, the full anti-cancer potential of cisplatin remains underutilized due to treatment induced toxicities, particularly nephrotoxicity. A signal cisplatin dose in the range of 50–100 mg/m2 results in nephrotoxicity in about 30% of the patients and 50–70% of the patients receiving 5-day consecutive cisplatin at 15–20 mg/m2/day developed nephrotoxicity. Therefore, nephrotoxicity is a particularly limiting factor for cancer patients to remain in the effective treatment due to the acute kidney injury (AKI). Moreover, patients recovered from acute kidney injury from supportive measures have a 25% increase in risk of developing chronic kidney disease and a 50% increase in 10-year mortality risk. Purpose Although cisplatin induced AKI is well recognized, the role of vasculature damage in cisplatin induced AKI remains to be explored. Accordingly, this study is focused on evaluating the role of vasculature preservation in cisplatin induced AKI. Methods Cisplatin induced AKI mouse model was generated by i.p. injecting 30 mg/kg cisplatin to C57BL/6 mice. Saline injection was served as control (n=3 per group). To evaluate blood vessel damage induced by cisplatin, mice were sacrificed 24 and 48 hours post injection. DyLight 488 labeled Lectin (Cat#: DL-1174, Vector Laboratories, Burlingame, CA) was injected at dose of 2 ml/kg 10 minutes before the sacrifice. To mitigate cisplatin induced AKI, rapamycin PFC nanoparticles at 1 ml/kg were i.v. injected 24 hours before cisplatin injection (n=10 per group). Realtime in vivo visualization on blood vessel damage was achieved by using intravital microscopy on the mesenteric microcirculation with regard to fluorescence PFC nanoparticle extravasation (i.v. injected right before the visualization) and then intestine was excised for fluorescence microscopic evaluation. Results As demonstrated in Figure 1A–C, cisplatin induced vascular injury happened no later than 24 hours post cisplatin injection and worsened at 48 hours (less green fluorescence). As shown in Figure 1D, all the mice received 1 ml/kg rapamycin PFC nanoparticles were survived 48 hours post cisplatin injection. However, only 60% of the mice survived at 48 hours in the group with saline injection (n=10 per group). The rapamycin PFC nanoparticles treatment resulted in better vascular preservation, therefore there is less fluorescence PFC nanoparticle (∼220 nm) extravasation shown less red signaling in Figure 1E, comparing Figure 1F. Western blot results suggested enhanced autophagy and attenuated inflammation through mTOR signaling could be the underlying therapeutic mechanism of rapamycin PFC nanoparticles (Figure 1G). Conclusions Preservation of vasculature could be beneficial in reducing cisplatin induced acute kidney injury. Funding Acknowledgement Type of funding sources: Other. Main funding source(s): NIH

Liquid biopsies for early diagnosis of brain tumours: in silico mathematical biomarker modelling.

Brain tumours are the biggest cancer killer in those under 40 and reduce life expectancy more than any other cancer. Blood-based liquid biopsies may aid early diagnosis, prediction and prognosis for brain tumours. It remains unclear whether known blood-based biomarkers, such as glial fibrillary acidic protein (GFAP), have the required sensitivity and selectivity. We have developed a novel in silico model which can be used to assess and compare blood-based liquid biopsies. We focused on GFAP, a putative biomarker for astrocytic tumours and glioblastoma multi-formes (GBMs). In silico modelling was paired with experimental measurement of cell GFAP concentrations and used to predict the tumour volumes and identify key parameters which limit detection. The average GBM volumes of 449 patients at Leeds Teaching Hospitals NHS Trust were also measured and used as a benchmark. Our model predicts that the currently proposed GFAP threshold of 0.12 ng ml−1 may not be suitable for early detection of GBMs, but that lower thresholds may be used. We found that the levels of GFAP in the blood are related to tumour characteristics, such as vasculature damage and rate of necrosis, which are biological markers of tumour aggressiveness. We also demonstrate how these models could be used to provide clinical insight.

Neuroprotective Effects of Electrical Stimulation Following Ischemic Stroke in Non-Human Primates.

Brain stimulation has emerged as a novel therapy for ischemic stroke, a major cause of brain injury that often results in lifelong disability. Although past works in rodents have demonstrated protective effects of stimulation following stroke, few of these results have been replicated in humans due to the anatomical differences between rodent and human brains and a limited understanding of stimulation-induced network changes. Therefore, we combined electrophysiology and histology to study the neuroprotective mechanisms of electrical stimulation following cortical ischemic stroke in non-human primates. To produce controlled focal lesions, we used the photothrombotic method to induce targeted vasculature damage in the sensorimotor cortices of two macaques while collecting electrocorticography (ECoG) signals bilaterally. In another two monkeys, we followed the same lesioning procedures and applied repeated electrical stimulation via an ECoG electrode adjacent to the lesion. We studied the protective effects of stimulation on neural dynamics using ECoG signal power and coherence. In addition, we performed histological analysis to evaluate the differences in lesion volume. In comparison to controls, the ECoG signals showed decreased gamma power across the sensorimotor cortices in stimulated animals. Meanwhile, Nissl staining revealed smaller lesion volumes for the stimulated group, suggesting that electrical stimulation may exert neuroprotection by suppressing post-ischemic neural activity. With the similarity between NHP and human brains, this study paves the path for developing effective stimulation-based therapy for acute stroke in clinical studies.

Staging cardiac damage in patients with aortic regurgitation.

The impact of "downstream" pathophysiological cardiac consequences in aortic regurgitation patients were not well established. The aim of our study was to validate a staging system built for severe aortic stenosis in a large real-world cohort of aortic regurgitation (AR) patients, evaluating the prevalence of different stages of cardiac damage and assess its prognostic impact. Clinical, echocardiographic and outcome data of patients with moderate-severe AR who underwent transthoracic echocardiography between January/2014 and September/2019 were retrospectively analysed. Patients were classified according to the extent of cardiac damage: Stage 0 (no cardiac damage), Stage 1 (left ventricular damage), Stage 2 (mitral valve or left atrial damage), Stage 3 (tricuspid valve or pulmonary artery vasculature damage) and Stage 4 (right ventricular damage). The primary endpoint was all-cause mortality. A total of 571 patients (median age 73, 51% male) were enrolled: Stage 0 (14.0%), Stage 1 (21.5%), Stage 2 (49.2%), Stage 3 (12.3%) and Stage 4 (3.0%). Median follow-up time was 39.5months (IQR 22.2 to 61.0). At the end of follow-up, cumulative death was significantly higher in more advanced disease stages (log-rank p < 0.001). On multivariable analysis, Stage 3-4 was associated with increased risk of all-cause mortality (HR 3.20; 95% CI 1.48-6.93; p = 0.003). Our study suggests that extra-valvular damage is common in patients with significant AR and that a staging system developed for aortic stenosis also provides prognostic information in these patients. This staging system may be helpful for clinical decision-making regarding the time of valvular intervention.

Association of Respiratory Tract Infections causing Alterations in Lung Parenchyma and Pulmonary Vasculature with Body Electrolyte Imbalance

Aim: The assessment of serum electrolytes at the time of initial presentation of the patient with respiratory tract infection possibly causing lung parenchyma and pulmonary vasculature damage and serial monitoring during the stay could be beneficial in order to determine when and how to take remedial action when necessary. Methodology: A non-probability sampling was done on 139 subjects with suspected respiratory tract infection. For confirmation, culture, MTB PCR, COVID-19 testing was done to diagnose the nature of infection. Serum electrolytes were tested on chemical analyses Alinity instrument. Results: Most common infections found were COVID-19 and bacterial (n=59) collectively in a co-morbid state. Mycobacterium tuberculosis and fungal infections were also found in (n=8) each. Electrolytes imbalance was markedly observed in high prevalence amongst Tuberculosis and COVID-19 patients but also showed significant association with other respiratory investigated infections. Conclusion: A robust association of electrolyte imbalance was found in all cases presented with upper or lower respiratory tract infections. Keywords: Respiratory tract infections, electrolyte imbalance, Covid-19, MTB, Sodium, Potassium

COVID‐19 Increases Markers of Endothelial Damage in Normotensive Pregnant Women

COVID‐19 (CV19) is a disease caused by SARS‐CoV‐2 and has caused a pandemic since discovered in 2019, resulting in over 5 million deaths worldwide. The impact of CV19 on pregnancy are still under investigation and are potentially long‐lasting. However studies have reported that pregnant women affected by CV19 have an increased risk of preterm delivery, pregnancy complications, organ failure, respiratory distress, increased inflammation, increased hospitalization and poor fetal outcomes compared to women who do not contract CV19 during pregnancy. Outside of the pregnant literature, data reports that comorbidities associated with CV19 are associated with inflammation and hypertension (i.e. endothelial damage, kidney damage). In the current study we investigated the relationship between endothelial damage and CV19, hypothesizing that women who contract CV19 during pregnancy will have an increase in endothelial damage and poor birth outcomes. Women were enrolled into an IRB approved study and whole blood was collected at the time of hospital admission for delivery. Pregnant women (n=7/group) were grouped into one of three categories: Normotensive with no history of CV19, early (eCV19) a positive CV19 test > 10 weeks prior to delivery or term (tCV19) a positive CV19 test < 10 weeks prior to delivery. After centrifugation, plasma and serum was frozen at ‐20ºC for future use. Endothelial cells (HUVECs) were exposed to 10% serum for 24 hours, followed by media collection after an additional 24hr serum‐free media exposure. Circulating ET‐1 was extracted from plasma samples and ET‐1, sVCAM and ICAM were assayed via ELISA from both circulating and media samples. Data are expressed as mean±SEM. Comparisons between groups were analyzed via ANOVA followed by post‐hoc analysis. P < 0.05 was considered statistically significant. Endothelin is increased plasma of eCV19 (7.08±0.31,p<0.0001) and tCV19 (6.82±0.25,p<0.0001) normotensive women when compared to normotensive women (0.94±0.22) without the CV19 virus. SVCAM is also increased in plasma of eCV19 (1338±69.37, p<0.0001) and tCV19 (1355±36.18, p<0.0001) when compared to normotensive women without CV19 (75.51±4.67). There is no significance across groups in ICAM measurement in plasma from these women p=0.9024. There is also an increase in endothelin 1 measured in HUVEC media in eCV19 normotensive women (10.10±4.04, p=0.0318) when compared to normotensive women without CV19 (2.05±0.64) and an increase in SVCAM in eCV19 (72.65±5.69, p=0.0008) and tCV19 (50.78±10.68, p=0.002) normotensive women when compared to non‐CV19 normotensive women (4.94±0.86). There was no significance in birth outcomes as Gestational Age at delivery (37.79‐38.92 weeks, p=0.72), baby weight (2880‐3534 grams, p=0.33), systolic blood pressure (118.7‐123.1 mmHg, p=0.43) across groups. Results above shows that despite all women being normotensive, CV19 causes an increase in endothelial dysfunction regardless of when a patient contracted the virus. These results suggests that CV19 causes vasculature damage as endothelial damage markers are increased in both normotensive women who contracted the virus early and late as opposed to normotensive women without the virus. Despite all women being normotensive; CV19 is shown to be causing a direct effect on vasculature causing increases in endothelial damage markers.

Rhegmatogenous retinal detachment induces more severe macular capillary changes than central serous chorioretinopathy

To investigate hemodynamic changes in macula-off rhegmatogenous retinal detachment (RRD) and its impact on visual prognosis by comparing with central serous chorioretinopathy (CSC). Using optical coherence tomography angiography (OCTA), vascular density in the superficial capillary plexus and deep capillary plexus (DCP) was retrospectively compared with that in contralateral unaffected eyes in macula-off RRD and CSC eyes. In RRD eyes, pre- and postoperative ultra-widefield (UWF) fluorescein angiography (FA) were obtained to analyze vascular changes. In OCTA, both macula-off RRD and CSC eyes showed less density in macular DCP, compared to the unaffected fellow eyes. Compared to CSC, eyes affected by macula-off RRD showed a reduction in DCP vascular density and an increase in foveal avascular zone area, although it had a much shorter macular detachment period. In macula-off RRD, less density of DCP was strongly correlated with longer duration of detachment, greater ellipsoid zone disruption, and poor visual recovery. In UWF-FA, detached retina showed capillary hypoperfusion, venous stasis and leakage, which were improved after reattachment. In conclusion, macular capillary loss of flow, which was associated with photoreceptor disruption, correlated with duration of detachment in RRD. Early reattachment and reperfusion are required for minimizing macular vasculature and photoreceptor damage in macula-off RRD.

BCG vaccination provides protection against IAV but not SARS-CoV-2

Since the vast majority of species solely rely on innate immunity for host defense, it stands to reason that a critical evolutionary trait like immunological memory evolved in this primitive branch of our immune system. There is ample evidence that vaccines such as bacillus Calmette-Guérin (BCG) induce protective innate immune memory responses (trained immunity) against heterologous pathogens. Here we show that while BCG vaccination significantly reduces morbidity and mortality against influenza A virus (IAV), it fails to provide protection against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). In contrast to IAV, SARS-CoV-2 infection leads to unique pulmonary vasculature damage facilitating viral dissemination to other organs, including the bone marrow (BM), a central site for BCG-mediated trained immunity. Finally, monocytes from BCG-vaccinated individuals mount an efficient cytokine response to IAV infection, while this response is minimal following SARS-CoV-2. Collectively, our data suggest that the protective capacity of BCG vaccination is contingent on viral pathogenesis and tissue tropism.

Development of Novel Tetrapyrrole Structure Photosensitizers for Cancer Photodynamic Therapy.

The effectiveness of photodynamic therapy (PDT) is based on the triad effects of photosensitizer (PS), molecular oxygen and visible light on malignant tumors. Such complex induces a multifactorial manner including reactive-oxygen-species-mediated damage and the killing of cells, vasculature damage of the tumor, and activation of the organism immunity. The effectiveness of PDT depends on the properties of photosensitizing drugs, their selectivity, enhanced photoproduction of reactive particles, absorption in the near infrared spectrum, and drug delivery strategies. Photosensitizers of the tetrapyrrole structure (porphyrins) are widely used in PDT because of their unique diagnostic and therapeutic functions. Nevertheless, the clinical use of the first-generation PS (sodium porfimer and hematoporphyrins) revealed difficulties, such as long-term skin photosensitivity, insufficient penetration into deep-seated tumors and incorrect localization to it. The second generation is based on different approaches of the synthesis and conjugation of porphyrin PS with biomolecules, which made it possible to approach the targeted PDT of tumors. Despite the fact that the development of the second-generation PS started about 30 years ago, these technologies are still in demand and are in intensive development, especially in the direction of improving the process of optimization split linkers responsive to input. Bioconjugation and encapsulation by targeting molecules are among the main strategies for developing of the PS synthesis. A targeted drug delivery system with the effect of increased permeability and retention by tumor cells is one of the ultimate goals of the synthesis of second-generation PS. This review presents porphyrin PS of various generations, discusses factors affecting cellular biodistribution and uptake, and indicates their role as diagnostic and therapeutic (theranostic) agents. New complexes based on porphyrin PS for photoimmunotherapy are presented, where specific antibodies are used that are chemically bound to PS, absorbing light from the near infrared part of the spectrum. Additionally, a two-photon photodynamic approach using third-generation photosensitizers for the treatment of tumors is discussed, which indicates the prospects for the further development of a promising method antitumor PDT.

Optical angiography for diabetes-induced pathological changes in microvascular structure and function: An overview

Diabetes mellitus (DM) is a kind of metabolic disorder characterized by chronic hyperglycemia and glucose intolerance due to absolute or relative lack of insulin, leading to chronic damage of vasculature within various organ systems. These detrimental effects on the vascular networks will result in the development of various diseases associated with microvascular injury. Modern optical imaging techniques provide essential tools for accurate evaluation of the structural and functional changes of blood vessels down to capillaries level, which can offer valuable insight on understanding the development of DM-associated complications and design of targeted therapy. This review will briefly introduce the DM-induced structural and functional alterations of vasculature within different organs such as skin, cerebrum and kidneys, as well as how novel optical imaging techniques facilitate the studies focusing on exploration of these pathological changes of vasculature caused by DM both in-vivo and ex-vivo.

Dietary alcohol exacerbates the hepatotoxicity induced by pyrrolizidine alkaloids: Hazard from food contamination

Pyrrolizidine alkaloids (PAs) are the most common plant-derived toxins with emerging evidence to contaminate soil, water, nearby plants and derived food products. Outbreaks of human poisoning cases, due to the ingestion of PA-contaminated food, have been reported in various countries including Ethiopia. This study first investigated the contamination of PAs in retail honey in Ethiopia. A striking 77% of honey samples (27/30) were found to contain PAs with the content ranging over 1.5–323.4 μg/kg. Notably, these PAs were also found as contaminants in mead, an alcoholic beverage made from local honey, indicating the transfer of PAs from the primarily contaminated honey into mead. Further toxicological examinations revealed that long-term PA exposure caused vasculature damage, fibrosis, and steatosis in mouse livers, and co-exposure to dietary alcohol exacerbated the PA-induced chronic hepatotoxicity. Furthermore, the study revealed that moderate alcohol intake did not affect the initiation mechanism (hepatic cytochrome P450-mediated bioactivation) of PA-induced hepatotoxicity but significantly disturbed hepatic glutathione homeostasis, thereby increasing oxidative stress in mouse liver and enhancing PA-induced hepatotoxicity. Our findings exemplify the carry-over of PA contamination through the food chain. Precautionary interventions are warranted on the hazardous effects of dietary exposure to PAs, particularly with concomitant alcohol consumption.

Staging cardiac damage in aortic valve disease: one size fits all?

Abstract Background Nowadays, in patients with aortic regurgitation (AR), aortic valve surgery is indicated when severe and symptomatic or those with depressed LVEF. However, clinical outcomes of patients with significant aortic regurgitation are not influenced by these factors only. Recently, a new staging system for severe aortic stenosis has been proposed by Généreux on the basis of the extent of anatomic and functional cardiac damage. If this model could be applicable to an unselected significant AR population has not been tested. Purpose The aim of our study was to evaluate the prevalence of the different stages of extra-aortic valvular cardiac damage by the application of Généreux staging and its impact on prognosis in a large, real world cohort of significant AR patients. Methods This study retrospectively analysed the clinical, Doppler echocardiographic and outcome data in patients with grade III or greater AR between January 2014 and September 2019. According to the extent of cardiac damage on echocardiography, patients were classified as Stage 0 (no cardiac damage), Stage 1 (left ventricular damage), Stage 2 (mitral valve or left atrial damage), Stage 3 (tricuspid valve or pulmonary artery vasculature damage) or Stage 4 (right ventricular damage). Exclusion criteria were severe aortic stenosis and previous valve repair or replacement. The primary end-point was all-cause mortality. Results A total of 572 patients, aged 70.1±13.9 years, 294 (51.3%) men were enrolled. One third of patients were in NYHA I. Based on the proposed classification, 82 patients (14.3%) were classified in stage 0, 130 (22.7%) in stage 1, 276 (48.2%) in stage 2, 68 (11.8%) in stage 3 and 17 (3.0%) in stage 4. Median follow-up time was 3.3±1.9 years. There was a progressive increase in mortality rates according to staging: 8.5% in stage 0, 10.8% in stage 1, 24.9% in stage 2, 42.6% in stage 3 and 52.9% in stage 4 (p&amp;lt;0.001). On multivariable analysis, the extent of cardiac damage was independently associated with excess mortality (HR 1.69, 95% CI 1.29 to 2.21) Conclusion Our study demonstrated that this new staging system studied for aortic stenosis also provides increased prognostic value to patients with significant aortic regurgitation. This staging system can be helpful to identify the degree of extra-aortic valvular cardiac damage and to optimize the time of valvular intervention. Further prospective studies are needed to confirm the benefit of the applicability of this model in clinical practice. Funding Acknowledgement Type of funding sources: Public hospital(s). Main funding source(s): Centro Hospitalar Vila Nova de Gaia / Espinho Distribution of stages of cardiac damageSurvival analysis according to stage

The key role of gut–liver axis in pyrrolizidine alkaloid-induced hepatotoxicity and enterotoxicity

Pyrrolizidine alkaloids (PAs) are the most common phytotoxins with documented human hepatotoxicity. PAs require metabolic activation by cytochromes P450 to generate toxic intermediates which bind to proteins and form protein adducts, thereby causing cytotoxicity. This study investigated the role of the gut–liver axis in PA intoxication and the underlying mechanisms. We exposed mice to retrorsine (RTS), a representative PA, and for the first time found RTS-induced intestinal epithelium damage and disruption to intestinal barrier function. Using mice with tissue-selective ablation of P450 activity, we found that hepatic P450s, but not intestinal P450s, were essential for PA bioactivation. Besides, in RTS-exposed, bile duct-cannulated rats, we found the liver-derived reactive PA metabolites were transported by bile into the intestine to exert enterotoxicity. The impact of gut-derived pathogenic factors in RTS-induced hepatotoxicity was further studied in mice with dextran sulfate sodium (DSS)-induced chronic colitis. DSS treatment increased the hepatic endotoxin level and depleted hepatic reduced glutathione, thereby suppressing the PA detoxification pathway. Compared to RTS-exposed normal mice, the colitic mice displayed more severe RTS-induced hepatic vasculature damage, fibrosis, and steatosis. Overall, our findings provide the first mode-of-action evidence of PA-induced enterotoxicity and highlight the importance of gut barrier function in PA-induced liver injury.