Vascular Necrosis Research Articles

Potential Role of Bone Scintigraphy in the Diagnosis of Calciphylaxis.

Nonosseous abnormalities are often seen on bone scans and can be related to a wide variety of pathology ranging across vascular, infection, and inflammatory etiology. Diffuse soft tissue radiotracer uptake on bone scans is typically attributed to renal or metabolic derangements. Calciphylaxis is the deposition of calcium in small blood vessels, skin, and other organs leading to vascular obstruction and skin necrosis. It is a rare disorder with unknown pathophysiology. Diagnosis of calciphylaxis is challenging and requires an interdisciplinary approach including clinical findings, laboratory results, medical imaging, and skin biopsy. An early diagnosis is important as the disease is associated with high morbidity and mortality. The purpose of this review article is to highlight the role of bone scintigraphy in the evaluation of calciphylaxis and to correlate the findings with other imaging modalities and histopathology.

Trimethylamine N-oxide aggravates vascular permeability and endothelial cell dysfunction under diabetic condition: in vitro and in vivo study.

To provide the direct evidence for the crucial role of trimethylamine N-oxide (TMAO) in vascular permeability and endothelial cell dysfunction under diabetic condition. The role of TMAO on the in vitro biological effect of human retinal microvascular endothelial cells (HRMEC) under high glucose conditions was tested by a cell counting kit, wound healing, a transwell and a tube formation assay. The inflammation-related gene expression affected by TMAO was tested by real-time polymerase chain reaction (RT-PCR). The expression of the cell junction was measured by Western blotting (WB) and immunofluorescence staining. In addition, two groups of rat models, diabetic and non-diabetic, were fed with normal or 0.1% TMAO for 16wk, and their plasma levels of TMAO, vascular endothelial growth factor (VEGF), interleukin (IL)-6 and tumor necrosis factor (TNF)-α were tested. The vascular permeability of rat retinas was measured using FITC-Dextran, and the expression of zonula occludens (ZO)-1 and claudin-5 in rat retinas was detected by WB or immunofluorescence staining. TMAO administration significantly increased the cell proliferation, migration, and tube formation of primary HRMEC either in normal or high-glucose conditions. RT-PCR showed elevated inflammation-related gene expression of HRMEC under TMAO stimulation, while WB or immunofluorescence staining indicated decreased cell junction ZO-1 and occludin expression after high-glucose and TMAO treatment. Diabetic rats showed higher plasma levels of TMAO as well as retinal vascular leakage, which were even higher in TMAO-feeding diabetic rats. Furthermore, TMAO administration increased the rat plasma levels of VEGF, IL-6 and TNF-α while decreasing the retinal expression levels of ZO-1 and claudin-5. TMAO enhances the proliferation, migration, and tube formation of HRMEC, as well as destroys their vascular integrity and tight connection. It also regulates the expression of VEGF, IL-6, and TNF-α.

Histopathological assessment of Atlantic salmon exposed to calcium oxide particles: a controlled clinical study

Particulate calcium oxide (CaO) has been successfully used for the control of sea urchin and starfish populations. Recent studies have proven its efficacy in killing planktonic copepods, making it a promising and cheap option for salmon louse control in the Atlantic salmon (Salmo salar) farming industry. Additionally, preliminary laboratory tests demonstrated that exposure to 0.2 g/L–0.6 g/L of fine CaO particles induced significant mortality of salmon louse (Lepeophtheirus salmonis) in the planktonic stages. The present study investigated the effects of water treatment with fine CaO particles [0.2 g/L or 127.4 g/m2; 0.1 mm–0.3 mm], conducted twice a week for three consecutive weeks at 5°C and 12°C on Atlantic salmon post-smolt in a flow-through system. The study compared mortality rates and histopathology of skin, eyes, gills, and intestines between treated and untreated control salmon. The results indicated that CaO exposure did not induce fish mortality or histopathological damages in the skin, eyes, or intestines. Although there were no significant effects of CaO exposure on gill inflammation or hyperplasia, the exposure did increase the occurrence of gill vascular injuries and necrosis in small portions of the tissue (< 10% of the respiratory gill tissue in most of the samples) by 60% and 35%, respectively. The effect was not modulated by temperature or time. The results indicate that the CaO treatment protocol used in this study is not lethal to Atlantic salmon and is safe for most analyzed tissues. Nonetheless, the induction of vascular damage and necrosis in the gill, albeit in small tissue portions, emphasizes the need for further research. It is crucial to develop a protocol that avoids such lesions, ensuring the secure application of CaO as an antiparasitic treatment in salmon aquaculture.

Preventive and therapeutic effects of the peel powder of P. granatum in a rat sepsis model

The aim of the present study was to evaluate the treatment potential of Punica granatum L. peel powder in an experimentally induced sepsis model in rats. Sepsis was induced in 10-week-old, male, Wistar Albino (n=24) rats. The animals were divided into four groups: Sham-operated (S) Group, Control (C) Group, Treatment-1 (T1) Group, and Treatment-2 (T2) Group. To induce the sepsis model, the cecal ligation and puncture procedure was performed. The P. granatum peel powder (200 mg/kg; per os) was applied one hour before (T1) and 10 hours after (T2) surgery in a volume of 2 mL. At the end of the experimental procedure, microbial and histopathological analyses were performed. The histopathological scores on the liver, lungs, heart, kidney, spleen, and pancreas were evaluated. Escherichia coli, Staphylococcus aureus and E. coli + S. aureus were isolated from blood cultures. Severe bacteria were detected in the blood of the group C animals. It was found that there were fewer bacteria in groups T1 (n=2) and T2 (n=4) compared with group C. There were no lesions in the pancreas tissues of any groups. Vascular changes, degeneration, and necrosis were common in the organs in all cases of group C compared to group S. The findings in group T1 were similar to those in group C, however, it was seen in fewer animals. It was determined that there was a general improvement in group T2, and in addition the existing lesions were moderate in severity. In conclusion, P. granatum L. peel powder prevented CLP-induced lung injury in experimental rats. Thus, P. granatum L. peel powder may be an alternative therapeutic agent against lung tissue injury induced by sepsis. The recovery from inflammation was better in group T2 than in the other groups. According to the results of the current study, P. granatum peel powder was found to be effective in the treatment of sepsis with antimicrobial and anti-inflammatory functions.

Vascular growth factors of the placenta, apoptosis of blood neutrophils in pregnant women with post-COVID syndrome

Research objectives: to determine the activity of vascular placental growth factors, apoptosis and necrosis of neutrophils in the blood of pregnant women with post-COVID syndrome.Methods and materials. 30 pregnant women (the main group) with SARS-CoV-2, positive Ig A, M or G to SARS-CoV-2 at 30–34 weeks of pregnancy took part in the study. The control group consisted of 30 women with physiological pregnancy.Blood neutrophils were studied by flow cytofluorometry. The proportion of neutrophils that were at the stage of apoptosis and necrosis was determined. The correlation of these indicators with the uncomplicated course of the gestational process during the physiological course of pregnancy and with the development of gestational complications against the background of post-COVID syndrome was determined.In the third trimester of pregnancy, we collected blood samples to determine the effect of SARS-CoV-2 infection on placental angiogenesis. We evaluated a panel of biomarkers: vascular endothelial growth factor (VEGF), placental growth factor PlGF, and interleukin-32α (IL-32α).Results. In pregnant women of the main group with a complicated course of pregnancy accompanied by post-COVID syndrome, apoptosis was at the level of 24.30 ± 0.50% of neutrophil cells, which was significantly different from the control group (4.45 ± 0.25%) (р < 0.001). The level of late apoptosis, necrosis of neutrophils increased 4 times – from 4.20 ± 0.65 to 16.80 ± 0.54% (р < 0.001). The concentration of IL-32α in pregnant women of the control group was 67.27 ± 5.63 pg/ml. Post-COVID syndrome caused an increase in this indicator in the main group by 2.8 times compared to the control group (188.36 ± 25.22 pg/ml) (p < 0.001). In the III trimester, the concentration of VEGF reached maximum values in the main group and was 192.20 ± 10.02 pg/ml, which is 2 times higher than in the control group at the same time (95.30 ± 5.65 pg/ml), (p < 0.001). In the control group the level of PIGF at full-term pregnancy was 144.53 ± 15.55 pg/ml. In women with postpartum syndrome, PIGF significantly decreased and was 43.92 ± 4.81 pg/ml, which was only 30% of the PIGF value in women with an uncomplicated pregnancy (p < 0.001).Conclusions. The development of metabolic disorders and apoptotic changes in the placental tissue are confirmed at the morphological level in the form of destructive and necrotic changes in the microcirculatory channel of the placenta. An increase in the level of annexin-positive neutrophils and activation of the degree of neutrophil necrosis is accompanied by fetoplacental dysfunction, by violation of the balance of placental growth factors and is an important marker for predicting of fetal growth retardation in pregnant women with post-COVID syndrome.

Small extracellular vesicles from failing heart accelerate tumour growth

Abstract Background Heart failure (HF) is associated with an increased incidence of cancer independently of shared risk factors. The mechanism behind this link is not entirely clear. Purpose We aimed to test the hypothesis that cardiac extracellular vesicles (cEVs) contribute to the link between HF and cancer. In addition, we aimed to determine if heart failure (HF) therapy will reduce the tumour-promoting effects of HF. Methods We used a mouse model of heterotopic lung cancer tumours and post-myocardial infarction (MI) HF. To determine the role of cEVs in tumour growth, we focused on cardiac mesenchymal stromal cells (cMSCs). First, we isolated cMSCs from mice hearts 10d after MI or sham MI and purified cMSC-EVs from the conditioned medium using size exclusion chromatography. Then, we analyzed cMSC-EV characteristics, cargo, and neoplastic effects on lung cancer cells. We also used cMSC-EV transfer to assess EV biodistribution and its effect on tumour growth. Next, we tested the effects of EV depletion on tumour growth. Finally, we used spironolactone to determine if HF therapy would attenuate the harmful interaction between HF and cancer. Results Mice with HF developed larger lung tumours than sham-MI mice during 28 days of follow-up. Failing hearts, particularly cMSCs, produced twice more EVs with neoplastic properties than non-failing hearts (Fig-A). Proteomics analysis revealed unique protein profiles of cMSC-EVs from failing hearts (Fig-B). cMSC-EVs from the failing hearts harbored higher quantities of tumour-promoting cytokines such as periostin, osteopontin, vascular endothelial growth factor, interleukin-6 and tumour necrosis factor-alpha; and tumorigenic microRNAs (miR) such as miR-221 and mir-21. Transfer of labeled cMSC-EVs from the failing heart into the systemic circulation of tumour-bearing mice targeted tumour cells and accelerated tumour growth (Fig-C). Moreover, EV depletion reduced the tumour-promoting effects of HF, and adoptive transfer of cMSC-EVs from failing hearts accelerated tumour growth in mice during systemic EV depletion (Fig-D). Finally, we used spironolactone to determine if HF therapy would inhibit the tumour trophic effects of HF. Spironolactone attenuated LV dilatation and improved global longitudinal strain following MI. Subsequently, spironolactone treatment reduced the number of EVs secreted by cMSCs from failing hearts by 26% and reduced the accelerated tumour growth after MI and HF (Fig E). Notably, spironolactone treatment did not reduce the volume of the tumours in sham-MI mice, indicating that the anti-tumour effects of spironolactone were related to its effects on the faling heart. Conclusions Our results provide new insight into the link between heart failure and cancer. We show, for the first time, that cardiac extracellular vesicles promote tumour growth independently of other secreted factors. Moreover, heart failure therapy reduces the tumour-promoting effects of heart failure.

Pedunculagin and tellimagrandin-I stimulate inflammation and angiogenesis and upregulate vascular endothelial growth factor and tumor necrosis factor-alpha in vivo

Pedunculagin (PD) and tellimagrandin-I (TL), isolated from Myrciaria cauliflora seeds and Eucaliptus microcorys leaves, respectively, have attracted great attention owing to their relevant biological activities, such as antitumor, antioxidant, and hepatoprotective activities. This study investigated the angiogenic potential of PD and TL using a chick embryo chorioallantoic membrane (CAM) assay. Using the CAM assay, our results showed that both PD and TL promoted a significant increase in the number and caliber of blood vessels, the thickness of the CAM, and the presence of fibroblasts and inflammatory cells. Moreover, an increase of tumor necrosis factor-α and vascular endothelial growth factor was observed in the CAM treated with PD and TL, indicating the induction of angiogenic factors. Thus, the remarkable profile of PD and TL in inducing angiogenesis opens up new perspectives for their potential utilization in different therapeutic approaches involving neovascularization.

Three-dimensional printed titanium mesh combined with iliac cancellous bone in the reconstruction of mandibular defects secondary to ameloblastoma resection

BackgroundThe reconstruction of large mandibular defects is a challenge, and free vascularized bone flaps are most commonly used. However, the precision and symmetry of this repair are deficient, and patients have a risk of vascular embolism, flap necrosis, and donor site complications. Therefore, to explore an ideal alternative in mandibular reconstruction with high surgical accuracy and low complications is indispensable.MethodsSeven patients with recurrent or large-scope ameloblastoma were enrolled in this study. All patients were provided with a fully digital treatment plan, including the design of osteotomy lines, surgical guides, and three-dimensional printed titanium mesh for implantation. With the assistance of surgical guide, ameloblastomas were resected, and custom 3D printed titanium mesh combined with posterior iliac bone harvest was used in mandibular reconstruction. A comparison was made between the discrepant surgical outcomes and the intended surgical plan, as well as the average three-dimensional deviation of the mandible before and after the surgery. At the same time, the resorption rate of the implanted bone was evaluated.ResultsAll patients completed the fully digital treatment process successfully without severe complications. Image fusion showed that the postoperative contour of the mandible was basically consistent with surgical planning, except for a slight increase in the inferior border of the affected side. The mean three-dimensional deviation of the mandible between the preoperative and postoperative periods was 0.78 ± 0.41 mm. The mean error between the intraoperative bone volume and the digital planning bone volume was 2.44%±2.10%. Furthermore, the bone resorption rates of the harvested graft 6 months later were 32.15%±6.95%.ConclusionsThe use of digital surgical planning and 3D-printed templates can assist surgeons in performing surgery precisely, and the 3D-printed titanium mesh implant can improve the patient’s facial symmetry. 3D printed titanium mesh combined with posterior iliac cancellous bone graft can be regarded as an ideal alternative in extensive mandibular reconstruction.

The involvement of NETs in ANCA-associated vasculitis.

Anti-neutrophil cytoplasmic autoantibody (ANCA)-associated vasculitis (AAV) is a serious autoimmune disease that is characterized by vascular necrosis. The pathogenesis of AAV includes ANCA-mediated neutrophil activation, subsequent release of inflammatory cytokines and reactive oxygen species (ROS), and formation of neutrophil extracellular traps (NETs). Excessive NETs could participate not only in ANCA-mediated vascular injury but also in the production of ANCAs per se as autoantigens. Thus, a vicious cycle of NET formation and ANCA production is critical for AAV pathogenesis. Elucidating the molecular signaling pathways in aberrant neutrophil activation and NETs clearance systems will allow specific therapeutics to regulate these pathways. Currently, standard therapy with high doses of glucocorticoids and immunosuppressants has improved outcomes in patients with AAV. However, AAV frequently develops in elderly people, and adverse effects such as severe infections in the standard regimens might contribute to the mortality. Mechanistically, cytokines or complement factors activate and prime neutrophils for ANCA-binding; thus, C5a receptor blocker has garnered attention as potential replacement for glucocorticoids in clinical settings. Recent studies have demonstrated that receptor-interacting protein kinases (RIPK3) and cyclophilin D (CypD), which regulate cell necrosis, may be involved in ANCA-induced NETs formation. Meanwhile, targeting NETs clearance, including the addition of deoxyribonuclease I (DNase I) and macrophage engulfment, may improve vasculitis. In this review, we focus on the pathogenesis of NETs and discuss potential targeted therapies for AAV based on recent experimental evidence.

Downregulation of MicroRNA-29-3p Following Percutaneous Coronary Intervention: An Implication of YY1/IRAK1 Pathway in the Post-Vascular Injury Inflammation.

This study explored the expression of microRNA (miR)-29b-3p following percutaneous coronary intervention (PCI) and the implication of its downstream Yin Yang 1 (YY1)/interleukin (IL)-1 receptor-associated kinase 1 (IRAK1) pathway in post-vascular injury inflammation. Blood samples were collected for analysis of plasma miR-29b-3p from patients with acute coronary syndrome before surgery, 1 day after PCI, and 30 days after PCI. Lipopolysaccharide (LPS)-treated human coronary artery endothelial cells (HCAECs) were transfected with miR-29b-3p mimic/inhibitor or YY1 shRNA and underwent viability tests. Enzyme-linked immunosorbent assay was performed to detect the levels of soluble vascular cell adhesion molecule-1 (sVCAM-1), IL-1β, IL-6, and tumor necrosis factor (TNF)-α in serum and cell culture supernatant. Dual-luciferase reporter and RNA/chromatin immunoprecipitation were used to confirm the targeting relationships among miR-29b-3p, YY1, and IRAK1. A rat model of intraluminal injury of the common femoral artery was established to address the role of miR-29b-3p and relevant mechanisms. miR-29b-3p was lowly expressed, and sVCAM-1, IL-1β, IL-6, and TNF-α were upregulated 1 day after PCI and 24 h after LPS treatment. miR-29b-3p overexpression or YY1 knockdown alleviated LPS-induced inflammatory responses and improved the viability of HCAECs. miR-29b-3p inhibition aggravated LPS-induced inflammatory injury in HCAECs. miR-29b-3p bound to YY1 mRNA and inhibited the expression of YY1 protein. YY1 bound to the IRAK1 promoter and activated the transcription of IRAK1. Upregulation of miR-29b-3p suppressed the inflammatory response after intraluminal injury of the common femoral artery in rats. In conclusion, dysregulation of the YY1/IRAK1 pathway via miR-29b-3p downregulation may be implicated in post-vascular injury inflammation.

Moderate LMWH Anticoagulation Improves Success Rate of Hind Limb Allotransplantation in Mice.

The mouse hind limb model represents a powerful research tool in vascularized composite tissue allotransplantation, but its applicability is limited due to poor graft survival (62%-83%). Vascular thrombosis and massive hemorrhage are the major causes for these drop-outs. We hypothesize that because of better anticoagulation effect and lower risk of thrombocytopenia, application of low molecular weight heparin (LMWH) will minimize vascular complications and enhance graft and animal survival. Fifty allogeneic hind limb transplantations were performed (C57BL/6 to DBA/2 mice) using five different anticoagulation protocols. Bleeding and thromboembolic events were recorded macroscopically by postoperative hemorrhage and livid discoloration of the graft, respectively. Graft perfusion and survival were monitored daily by capillary-refill-time of graft toes within 2-3 seconds. Vascular congestion and tissue necrosis were examined by histological evaluation of hematoxylin-eosin-stained tissue sections. All transplantations were technically successful. Increase in thromboembolic events and a concomitant decrease in bleeding events were observed with the decreasing concentration of heparin in the perfusion solution. Although treatment of donor and recipient with low dose of LMWH could not reduce thromboembolic events, moderate dose effectively reduced these events. Compared with the poor outcome of graft perfusion with heparin alone, additional treatment of donor and recipient with low dose of LMWH improved graft and animal survival by 18%. Interestingly, animals treated with moderate dose of LMWH demonstrated 100% graft and animal survival. Treatment of donor and recipient mice with a moderate dose of LMWH prevents vascular complications and improves the outcome of murine hind limb transplants.

Prognostic value of S100A4 and Glypican-3 in hepatocellular carcinoma in cirrhotic HCV patients

AimsBoth S100A4 and Glypican-3 have been known to be engaged in HCC development and progression. This study aimed to evaluate both S100A4 and GPC3 expression in HCC tissues as a prognostic markers.MethodsTissues from 70 patients of HCC in cirrhotic HCV patients were evaluated by immunohistochemistry using antibodies against SA100A4 and GPC3 and compared with tumor-adjacent tissue (controls). All cases were followed for 40 months.ResultsGPC3 was more expressed in HCC (79%) than S100A4 (21%). S100A4 was more significantly expressed in cases showing metastasis, microscopic vascular emboli, necrosis, and grade III tumors. There was no relationship between overall survival and both S100A4 and GPC3. The only significant independent predictor for recurrence was decompensation (OR 3.037), while metastasis was significantly predicted by S100A4 expression (OR 9.63) and necrosis (OR 8.33).ConclusionS100A4 might be used as a prognostic marker for HCC, while GPC3 is a reliable marker of HCC diagnosis.

First report of Fusarium equiseti causing root and crown rot in tomato in Mexico.

Tomato (Solanum lycopersicum L.) is one of the most important crops in Mexico due to its economic and nutritional value. Among the main diseases in tomato production is Fusarium wilt, which can cause 60% production losses (Ascencio et al, 2008). Mixed infections of Fusarium species or other fungi genera, would increase disease severity. During April to May of 2021, tomato plants with more than 60 days old, were collected from the main production areas of Aguascalientes (22°03'46.5"N 102°05'17.4"W and 22°04'53.64"N 101°58'55.81"W) and Zacatecas (23°05'59.2"N 102°41'07.3"W and 22°16'52.1"N 102°00'11.8"W) Mexico states. Plants showed main root rot, vascular bundles necrosis with corky appearance, stem crown rot, and ascending yellowing. The main root and stem crown were cut in 0.25 cm2 pieces and disinfested in 2% NaClO for one minute, rinsed with distilled water two times, placed on acidified potato dextrose agar (PDA) medium, and incubated at 25 ± 2°C for 7 days. Characteristic Fusarium growths were purified by hyphal tip on PDA, subsequently pure strains were obtained by single-spore isolation method. Several fungi colonies were obtained, but we focused on the colonies that showed abundant aerial mycelium of white color and irregular growth, which turned yellowish to golden and brown color as it ages. Carnation leaf agar (CLA) medium were used for conidia and sporodochium development. Chains of terminal, intercalary and agglomerated chlamydospores with thick, rough brown walls of 18.9 (7.46) µm in diameter (n=120) were observed in the mycelium. Macroconidia with 5 to 7 septa were 30 to 75 (28.32) µm in long and 1.2 to 4.8 (3.2) µm in wide (n=72). Basal cell developed in foot-shape, apical cell was elongated and slightly curved, and some macroconidia had swollen midd-cell. Sporodochium was orange to brown in color and microconidia were absent (Figure 1). Two representative strains from each state, LCA-3.1 and EMA-1 from Aguascalientes and ECZ-4 and LRZ-6 from Zacatecas, were selected for DNA amplification of ITS, TEF-1α and RPB2 regions, with universal primers ITS1/ITS4, EF1/EF2 and 2-5F2/7cR (White et al.1990; O'Donnell et al. 1998, 2013). PCR products were sequenced by Psomagen, Inc. (USA). The sequences obtained showed 100% of similarity among themselves and within species of the Fusarium incarnatum-equiseti species complex (FIESC) with nucleotide NCBI accessions NR_121457 (Type material) for ITS and MW362069 for TEF-1α; and 99.28% with MN170399 for RPB2 in FUSARIOID-ID database. According to morphological (Leslie and Summerell, 2006) and molecular characteristics, isolates were identified as Fusarium equiseti (FIESC 14). The LCA-3.1 sequences were selected to be deposited in GenBank with accession numbers OM812801 (ITS), OM937108 (TEF-1α) and ON653596 (RPB2). Pathogenicity tests were performed twice, under greenhouse conditions in tomato seedlings of cv. Rio Grande. Five tomato seedlings were inoculated by root immersion method (Lopez et al, 2018) in a 1x106 spores/mL solution for 8 min, and transplanted to 1L pots with sterile peat. Five controls plants were immersed in sterile water. At 14 days after inoculation, a general plant decline and slower growth compared to the control plants were observed. Subsequently, plants showed root rot, vascular necrosis, and a brown ring in stem crown. Controls were symptomless. The fungi were re-isolated from symptomatic plants and were morphologically similar to the inoculated strains. Patel et al. (2017) described the pathogenic and toxic effects of F. equiseti on tomato, causing low seed germination, and low root and shoot growth. This is the first report of F. equiseti causing root and stem rot in tomato plants in Mexico.

Primer reporte de Fusarium oxysporum f. sp. niveum raza 1 como agente causal de la marchitez vascular de la sandía en México

Vascular wilt (Fusarium oxysporum f. sp. niveum [Fon]) is the main fungal disease of watermelon crops worldwide and can be caused by any of the four known races of this fungus (race 0, 1, 2 and 3). In Mexico, there are no reports of the presence of any of these races. The presence of plants with wilt and vascular necrosis has been observed in commercial watermelon plantations in the municipality of Hermosillo, Sonora, Mexico, mainly during the spring-summer growing seasons. The aim of this study was to identify the Fon race causing vascular wilt in watermelon plants in Hermosillo, Sonora. Three fungal isolates were collected from symptomatic plants. Based on their morphological characteristics, DNA amplification of the isolates (with specific primers Fon-1/Fon-2, FONSIX6F/FONSIX6R and FNR3-F/FNR3-R, which allow differentiating Fon race 1, 2 and 3) and inoculation into three differential watermelon cultivars, the three isolates (UESFON01, UESFON02 and UESFON03) were identified as F. o. f. sp. niveum race 1. To our knowledge, this is the first report where F. o. f. sp. niveum race 1 is identified as the causal agent of wilt, vascular necrosis and plant death in watermelon crops in Mexico.

A Bacterial Wilt Disease Caused by a New Bacterial Pathogen of Guar Belonging to the Genus Pantoea Identified in Texas

Guar ( Cyamopsis tetragonoloba [L.] Taub) or cluster bean is a legume plant that is cultivated mainly for its galactomannan (guar gum) extracted from the seed endosperm. In 2021, a disease outbreak among germplasms in the Texas A&M AgriLife Research Guar Breeding Program located in Vernon, Texas, was observed. Symptoms of the disease were similar to bacterial blight caused by Xanthomonas axonopodis pv . cyamopsidis and included black necrotic streaks on stems and petioles, vascular necrosis, wilting, and eventual death of affected plants. The disease incidence ranged from 75 to 90% in each of the affected germplasms. Diagnosis of affected germplasms consistently recovered bacteria that were identified on the basis of their 16S rRNA partial gene sequence as belonging to the genus Pantoea. Two independent pathogenicity assays and the satisfaction of Koch's postulate under greenhouse experimental conditions determined that the Pantoea isolates were the causal agent of the disease. Investigations of the disease etiology also led to the discovery that the Pantoea sp. can be seedborne. Additionally, although the disease symptoms share similarities with those of bacterial blight in guar, they differ in the absence of the large angular necrotic lesions on leaves that are characteristic of the later stage of the bacterial blight infection. However, infection by the Pantoea sp. similarly resulted in the eventual death of affected plants. Whole-genome pairwise alignments and multilocus sequence analysis of the isolate indicate that it is most closely related to P. vagans, but also distinct genetically, and tentatively designated as Pantoae sp. isolate B550. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Point-of-Care Tissue Oxygenation Assessment with SnapshotNIR for Alloplastic and Autologous Breast Reconstruction.

Patients receiving immediate alloplastic reconstruction after mastectomy or autologous reconstruction were enrolled. Preoperative, intraoperative, and postoperative images were taken of the flaps. StO2 and hemoglobin were measured at the following locations: superior and inferior breast, free flap skin paddle (when applicable), and un-operated control skin. Linear mixed effects model for repeated measurements was used to model measurements to estimate the area effect difference across time, time effect difference across area, and pairwise comparisons between two areas at each time point. Thirty-two breasts underwent alloplastic reconstruction; 38 breasts underwent autologous reconstruction. No enrollees developed skin necrosis. StO2 was highest after mastectomy and closure in alloplastic reconstructions. StO2 was observed to decline at follow-up in autologous reconstructions. Mean preoperative StO2 was highest in breasts that had previously undergone mastectomy and alloplastic reconstruction. The SnapshotNIR device detected normal spatial and temporal differences in tissue oxygenation over the operative course of alloplastic and autologous breast reconstruction. A multi-institutional, prospective clinical trial is needed to determine the sensitivity and specificity of this device for detecting skin flap necrosis.

Intraoperative Indocyanine Green Angiography Facilitates Flap Fenestration and Facial Organ Fabrication in Total Facial Restoration.

Because of a lack of effective measures to visualize flap vasculature and perfusion, flap fenestration and facial organ fabrication cannot be performed effectively, preventing the transition from two-dimensional coverage to restoration of the three-dimensional (3D) structure of facial organs. This study aimed to evaluate the efficacy of indocyanine green angiography (ICGA) in guiding flap fenestration and facial organ fabrication in total facial restoration. Ten patients with total facial scarring after burn injury were enrolled in the study. They were treated with preexpanded, prefabricated monoblock flaps for total face restoration. The openings of nostrils and oral and palpebral orifices, together with organ fabrication, were conducted under the guidance of intraoperative ICGA by hemodynamic evaluation of flap perfusion. Postoperative follow-up measures include vascular crisis, infection, flap necrosis, and aesthetic and functional recovery. The opening of facial organ orifices was performed at the stage of flap transfer in nine patients. To avoid damaging the major nourishing vessels, the left palpebral orifice was opened 8 days after the flap transfer in one patient, as observed by ICGA. Based on ICGA evaluation, the decision to perform additional vascular anastomosis before flap fenestration was made in six patients. Hemodynamic analysis of flap perfusion after fenestration revealed no significant change. Follow-up showed satisfactory aesthetic recovery and well-restored 3D structures of facial organs. This pilot study demonstrates how intraoperative ICGA can enhance the safety of flap fenestration, thereby transforming full facial restoration from the two-dimensional to the 3D realm by facilitating facial organ fabrication. Therapeutic, IV.

#2709 THE INVOLVEMENT OF NRF2-MEDIATED ANTIOXIDANT SYSTEM IN MYELOPEROXIDASE-POSITIVE ANTINEUTROPHIL CYTOPLASMIC ANTIBODY-ASSOCIATED VASCULITIS

Abstract Background and Aims Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a systemic autoimmune disease pathologically characterized by vascular necrosis with inflammation. The necrotic lesions consist of ANCA-mediated neutrophil extracellular traps (NETs). During AAV, activated neutrophils form NETs via reactive oxygen spices (ROS)s-mediated signaling pathway. The excessive NETs lead to endothelial vascular damage and serve as autoantigens against ANCAs in a vicious cycle. Although the role of NETs in AAV has been revealed, its treatment for targeting NETs remains unestablished. Since nuclear factor-erythroid 2-related factor 2 (Nrf2) regulates antioxidant proteins against oxidative stress and functions as a defense system, we investigated the role of Nrf2 in the development of AAV. Method In vitro, human neutrophils treated with the Nrf2 activator bardoxolone methyl (Bard) were stimulated with ANCA, and the effects on NETs formation and signaling pathways including intracellular ROS were evaluated by cell imaging and biochemical approaches. The effects of Nrf2 activators on NETs-induced cytotoxicity were examined using human renal glomerular endothelial cells (HRGECs). Nrf2 knockout or WT mouse-derived neutrophils were stimulated with anti-MPO antibody to induce NETs. In vivo studies, Bard was administered to ANCA-transfer AAV model or spontaneous AAV developing mice models (SCG/Kj). Renal function was assessed by biochemical analysis, and the expression of ROS and antioxidant protein NAD(P)H Quinone Dehydrogenase 1 (NQO1) in tissues was evaluated by fluorescent immunostaining and immunoblotting. The formation of NETs in the glomeruli was evaluated by immunostaining and glomerular vascular endothelial damage was assessed by the expression of CD31 and TUNEL positivity. To assess the role of Nrf2 in immunological abnormality during AAV, murine splenocytes were collected and flow cytometry were conducted using CD4, CD8, B220, MCHCII, CD11c, F4/80, Ly 6G, Ly6b, and CD11b antibodies. Moreover, to assess the genetic role of Nrf2 in AAV, we established SCG/Kj mice carrying a germ-line deletion of the Nrf2-genes. A spontaneous mouse model of vasculitis genetically deficient in Nrf2 was analyzed by the same parameters to evaluate the effect of Nrf2. N1 offspring mice were generated by crossing Nrf2-/- mice with SCG/Kj mice. These backcrossed mice were obtained by crossing Nrf2-/- mice to background SCG/Kj mice for three generations. The experimental group was mated to N4 heterozygous mice, and female mice in the experimental group were analyzed at 18 weeks. Results In vitro, pharmacological activation of Nrf2 suppressed ANCA-induced NETs via the upregulation of NQO1 and inhibition of ROS. While, Nrf2-deficienet neutrophils showed the massive NETs in response to ANCA, compared to WT neutrophils. Furthermore, Nrf2 activation directly protected endothelium from cellular injury caused by the exposure of ANCA-mediated NETs. In vivo, the pharmacological activation of Nrf2 ameliorated glomerulonephritis in two kinds of AAV models through the upregulation of antioxidant and inhibition of ROS-mediated NETs. In contrast, Nrf2 genetic deficiency exacerbated vasculitis in spontaneous AAV model. The activation of Nrf2 regulated the expansion of splenocytes in AAV mice. In particular, pan-T cells in spleen and the infiltration of Th17 cells in kidney were suppressed by Nrf2 activation. Conclusion During ANCA-stimulation in neutrophils, the activation of Nrf2 initiates antioxidant system and inhibits intracellular ROS, leading to the suppression of NETs formation. Nrf2 may be a novel therapeutic target for AAV.

Plasma proteome of Long-COVID patients indicates HIF-mediated vasculo-proliferative disease with impact on brain and heart function

AimsLong-COVID occurs after SARS-CoV-2 infection and results in diverse, prolonged symptoms. The present study aimed to unveil potential mechanisms, and to inform prognosis and treatment.MethodsPlasma proteome from Long-COVID outpatients was analyzed in comparison to matched acutely ill COVID-19 (mild and severe) inpatients and healthy control subjects. The expression of 3072 protein biomarkers was determined with proximity extension assays and then deconvoluted with multiple bioinformatics tools into both cell types and signaling mechanisms, as well as organ specificity.ResultsCompared to age- and sex-matched acutely ill COVID-19 inpatients and healthy control subjects, Long-COVID outpatients showed natural killer cell redistribution with a dominant resting phenotype, as opposed to active, and neutrophils that formed extracellular traps. This potential resetting of cell phenotypes was reflected in prospective vascular events mediated by both angiopoietin-1 (ANGPT1) and vascular-endothelial growth factor-A (VEGFA). Several markers (ANGPT1, VEGFA, CCR7, CD56, citrullinated histone 3, elastase) were validated by serological methods in additional patient cohorts. Signaling of transforming growth factor-β1 with probable connections to elevated EP/p300 suggested vascular inflammation and tumor necrosis factor-α driven pathways. In addition, a vascular proliferative state associated with hypoxia inducible factor 1 pathway suggested progression from acute COVID-19 to Long-COVID. The vasculo-proliferative process predicted in Long-COVID might contribute to changes in the organ-specific proteome reflective of neurologic and cardiometabolic dysfunction.ConclusionsTaken together, our findings point to a vasculo-proliferative process in Long-COVID that is likely initiated either prior hypoxia (localized or systemic) and/or stimulatory factors (i.e., cytokines, chemokines, growth factors, angiotensin, etc). Analyses of the plasma proteome, used as a surrogate for cellular signaling, unveiled potential organ-specific prognostic biomarkers and therapeutic targets.

Value of transrectal ultrasound-guided biopsy in endoscopy negative biopsy patients with rectal lesions

ObjectiveTo evaluate the value of transrectal ultrasound (TRUS) guided biopsy in endoscopy negative biopsy patients with rectal lesions.Methods150 endoscopy negative biopsy result rectal lesions adopted the transrectal ultrasound-guided biopsy. Based on whether the patients received contrast-enhanced ultrasound examination or not before the biopsies, all enrolled cases were divided into TRUS guided group and contrast-enhanced TRUS (CE-TRUS) guided group, and the safety and diagnostic performances were analyzed retrospectively.ResultsWe obtained adequate specimens in the majority of cases (98.7%,148/150); There were no complications identified in our study. 126 patients received contrast-enhanced TRUS examination before biopsy to evaluate vascular perfusion and necrosis. The sensitivity, specificity, PPV, NPV, and overall accuracy of all biopsies were 89.1%, 100%,100%, 70.4%, and 91.3% respectively; The sensitivity, specificity, PPV, NPV, and accuracy for TRUS-guided biopsy and CE-TRUS guided biopsy were 73.7%, 100%,100%, 50%,79.2% and 92.1%,100%,100%,75%,93.6% separately; The increase in correct diagnoses was significant (p < 0.05) between TRUS guided biopsy and CE-TRUS guided biopsy.ConclusionTRUS-guided biopsy is a reliable procedure that can be augmented by endoscopic biopsy techniques if the biopsy yields negative results. CE-TRUS might assist in the location of the biopsy and decrease sampling errors.