Capillary Loss Research Articles

FOXF1 Regulates Alveolar Epithelial Morphogenesis through Transcriptional Activation of Mesenchymal WNT5A.

Mutations in the FOXF1 (forkhead box F1) gene, encoding the mesenchymal FOX (forkhead box) transcription factor, are linked to alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV), a severe congenital disorder associated with the loss of alveolar capillaries and lung hypoplasia. Although proangiogenic functions of FOXF1 have been extensively studied, the role of FOXF1 in mesenchymal-epithelial signaling during lung development remains uncharacterized. Herein, we used murine lung organoids to demonstrate that the S52F FOXF1 mutation (found in patients with ACDMPV) stimulates canonical WNT/β-catenin signaling in type 2 alveolar epithelial cells (AEC2s), leading to increased proliferation of AEC2s and decreased differentiation of AEC2s into type 1 alveolar epithelial cells (AEC1s). Alveolar organoids containing Foxf1WT/S52F lung fibroblasts and wild-type epithelial cells grew faster on Matrigel and exhibited AEC2 hyperplasia. AEC2 hyperplasia and loss of AEC1s were found in the lungs of Foxf1WT/S52F embryos, a mouse model of ACDMPV. Activation of canonical WNT/β-catenin signaling in AEC2s of lung organoids and Foxf1WT/S52F mice was associated with decreased expression of noncanonical WNT5A (Wnt family member 5A) ligand in lung fibroblasts. Mechanistically, FOXF1 directly activates the Wnt5a gene transcription through an evolutionarily conserved +6320/+6326 region located in the first intron of the Wnt5a gene. Site-directed mutagenesis of the +6320/+6326 region prevented the transcriptional activation of the Wnt5a enhancer by FOXF1. Treatment with exogenous WNT5A ligand inhibited the effects of the S52F FOXF1 mutation on canonical WNT/β-catenin signaling in alveolar organoids, preventing aberrant AEC2 expansion and restoring differentiation of AEC1s. Activation of either FOXF1 or WNT5A may provide an attractive strategy to improve lung function in patients with ACDMPV.

Capillaroscopy, microangiopathy, and HIV. Descriptive study of capillaroscopy findings in HIV positive patients

In this study, we aim to evaluate microangiopathy in HIV positive patients by using capillaroscopy. To date, few studies have been published on the topic. Capillaroscopy may be a tool for early diagnosis of cardiovascular involvement in this patient population. Cross-sectional study with HIV positive patients >18 years. The enrolment period was set from January to June 2018. The following data were collected: demographic (sex, age), laboratory tests (duration of infection, CD4 cell count, CD4:CD8 ratio, coinfection with other viruses), antiretroviral treatment, dyslipidemia, and comorbidities (active smoking, alcoholism, high blood pressure, dyslipidaemia, diabetes, cardiopathy). The capillaroscopy and blood tests were performed simultaneously. The following alterations were evaluated in the capillaroscopy: congestion, tortuosity, haemorrhage, dilations, capillary loss, and presence of megacapillaries. One hundred and two patients were included; 73.5% were male, mean age was 40 years (SD: 10), and mean duration of infection 4.5 years (SD: 3.1). At diagnosis, mean CD4 cell count was 408/mm3 and CD4/CD8 ratio 0.4. A number of patients (14.7%) were coinfected with the hepatitis B virus; 31.3% were active smokers and 13.7% alcoholics. Capillaroscopy alterations were found in most study patients (93.1%): congestion (78.5%), tortuosity (77.5%), haemorrhage (13.8%), dilations (11.8%), capillary loss (5%), and megacapillaries (1%). Capillary tortuosity was associated with age and smoking; and haemorrhage with age, CD4, antiretroviral treatment, and hypertension. Prevalence of capillaroscopy alterations is high in HIV positive patients, particularly tortuosity and congestion. To the best of our knowledge, the later alteration has not been previously reported in this group of patients.

Swept‐source optical coherence tomography angiography in toxoplasmic retinochoroiditis

AbstractPurpose: To describe optical coherence tomography angiography (OCTA) findings in eyes with active and scarred toxoplasmic retinochoroiditis.Methods: OCTA scans in active (25 eyes) and scarred (17 eyes) retinochoroiditis were retrospectively reviewed at Fattouma Bourguiba University Hospital, Monastir, Tunisia.Results: In active lesions, OCTA findings included a non‐detectable flow signal area in retinal vascular plexuses and choriocapillaris in all 25 eyes (100%), an abnormal intraretinal vascular process in two eyes (8%), and an associated area of retinal flow deficit secondary to branch retinal artery occlusion in one eye (4%). In scarred lesions, OCTA findings included a flow deficit area in retinal vascular plexuses and choriocapillaris in all 17 eyes (100%), a visibility of larger deeper choroidal vessels at the level of choriocapillaris in nine eyes (53%), and a well‐defined intraretinal vascular network in one eye (5.9%). Peripapillary scars were associated on OCTA with wedge‐shaped loss of radial peripapillary capillaries with corresponding localized retinal nerve fibre layer defect and visual field loss.Conclusions: OCTA allows to non‐invasively detect retinal and choroidal vascular changes in active and scarred toxoplasmic retinochoroiditis.

Association between parapapillary choroidal microvasculature and prognosis of optic neuritis

AbstractPurpose: To identify peripapillary choroidal MvD in eyes with optic neuritis and its association with longitudinal changes in retinal nerve fibre layer (RNFL) and ganglion cell‐inner plexiform layer (GCIPL) thicknesses following diagnosis.Methods: A total of 48 eyes with optic neuritis was evaluated to identify the presence of peripapillary choroidal MvD, defined as a focal capillary loss with no visible microvascular network in choroidal layer, using optical coherence tomography (OCT) angiography (OCTA). Patients were divided based on the presence of MvD. OCT and standard automated perimetry (SAP) conducted at 1, 3 and 6 months follow‐up were analysed.Results: MvD was identified in 20 of 48 eyes (41.7%) with optic neuritis. MvD was most commonly found in the temporal quadrant (85.0%), and peripapillary retinal vessel density in the temporal quadrant was significantly lower in eyes with MvD (p = 0.012). At 6 months follow up, optic neuritis eyes with MvD showed significantly thinner RNFL in the temporal quadrant (p = 0.012) and GCIPL in all sectors (p = 0.007). No significant difference was noted in SAP parameters. The presence of MvD was significantly associated with thinner global GCIPL thickness at 6 months follow‐up (OR 0.905, 95% CI 0.832–0.984, p = 0.019).Conclusions: Optic neuritis showed peripapillary choroidal microvascular impairment in the form of MvD. MvD was associated with structural deterioration at macular GCIPL. Further studies are necessary to identify the causal relationship between microvascular impairment and retinal nerve fibre layer damage in optic neuritis.

Vision transformer assisting rheumatologists in screening for capillaroscopy changes in systemic sclerosis: an artificial intelligence model.

The first objective of this study was to implement and assess the performance and reliability of a vision transformer (ViT)-based deep-learning model, an 'off-the-shelf' artificial intelligence solution, for identifying distinct signs of microangiopathy in nailfold capilloroscopy (NFC) images of patients with SSc. The second objective was to compare the ViT's analysis performance with that of practising rheumatologists. NFC images of patients prospectively enrolled in our European Scleroderma Trials and Research group (EUSTAR) and Very Early Diagnosis of Systemic Sclerosis (VEDOSS) local registries were used. The primary outcome investigated was the ViT's classification performance for identifying disease-associated changes (enlarged capillaries, giant capillaries, capillary loss, microhaemorrhages) and the presence of the scleroderma pattern in these images using a cross-fold validation setting. The secondary outcome involved a comparison of the ViT's performance vs that of rheumatologists on a reliability set, consisting of a subset of 464 NFC images with majority vote-derived ground-truth labels. We analysed 17 126 NFC images derived from 234 EUSTAR and 55 VEDOSS patients. The ViT had good performance in identifying the various microangiopathic changes in capillaries by NFC [area under the curve (AUC) from 81.8% to 84.5%]. In the reliability set, the rheumatologists reached a higher average accuracy, as well as a better trade-off between sensitivity and specificity compared with the ViT. However, the annotators' performance was variable, and one out of four rheumatologists showed equal or lower classification measures compared with the ViT. The ViT is a modern, well-performing and readily available tool for assessing patterns of microangiopathy on NFC images, and it may assist rheumatologists in generating consistent and high-quality NFC reports; however, the final diagnosis of a scleroderma pattern in any individual case needs the judgement of an experienced observer.

The vascular protective effect of matrix Gla protein during kidney injury.

Matrix Gla protein (MGP) is a small secreted protein and requires vitamin K dependent γ-carboxylation for its function. MGP has been identified as a local inhibitor of vascular calcification because MGP-deficient mice die due to severe arterial calcification and resulting arterial rupture. Clinical trials revealed that reduction in active MGP predicts poor prognosis in patients due to cardiovascular complications. However, recent studies showed that MGP controls angiogenesis during development. MGP-deficient mice demonstrated abnormal hypervascularization and arteriovenous malformations in kidneys and other organs. This abnormal angiogenesis is largely caused by excessive expression of vascular endothelial growth factor-A (VEGF-A) and VEGF receptor-2 (VEGFR2). However, only a few studies have investigated the roles of MGP in tissue injury. We observed mesangial cell proliferation and mild interstitial fibrosis in addition to increased capillaries in kidneys of MGP-null mice even without injury. We also created a mouse model with kidney injury and found that kidney damage greatly increases MGP expression in peritubular capillary endothelial cells and tubular epithelial cells. Finally, our study showed that impairment of MGP expression aggravates peritubular capillary rarefaction and accumulation of collagen-producing myofibroblasts following kidney injury. Peritubular capillary damage induces capillary loss as well as trans-differentiation of vascular pericytes into myofibroblasts. These results indicate that MGP has the vascular protective effect in the injured kidney. Clinical trials have already started to test the efficacy of MGP activation to repair vascular calcification in patients with chronic kidney diseases. In this "Hypothesis and Theory" article, we discuss possible mechanisms by which MGP protects against vascular damage during tissue injury based on our experimental results and previous results from other research groups.

Improvement of microangiopathy after haematopoietic stem cell transplantation in systemic sclerosis.

Haematopoietic stem cell transplantation (HSCT) is a treatment option for patients with severe systemic sclerosis (SSc), but the efficacy of the procedure in remodelling the nailfold microvascular array is largely unknown. Therefore, this study aimed to evaluate the effect of HSCT on microangiopathy assessed through nailfold capillaroscopy (NC) and to compare the results with findings in patients receiving conventional immunosuppression. We included SSc patients with severe SSc and whose pre- and post-treatment NC images were available. Findings in patients treated with HSCT were compared with patients not treated with HSCT. Images were scored by two independent observers blinded for clinical data and treatment history. Capillary pattern was determined and semiquantitative scores from 0 (no changes) to 3 (>66% alterations per millimetre) were used to quantify the degree of specific microvascular characteristics. Changes in severity of microangiopathy between baseline and post-treatment were compared between groups. Images of 18 HSCT patients and 21 controls were scored. From baseline to follow-up, 33% of HSCT patients showed improvement from scleroderma pattern to normal NC, compared to 6% of controls (p=0.15). Pre- to post-treatment differences in semiquantitative scores showed significant improvement in HSCT patients compared to controls regarding capillary loss (-0.5 vs. 0.0, p<0.05) and disorganisation (-0.8 vs. 0.0, p<0.05). The degree of microangiopathy improved significantly in severe SSc patients treated with HSCT compared with patients receiving conventional immunosuppressive therapy.

Regulation of serine palmitoyl-transferase and Rac1–Nox2 signaling in diabetic retinopathy

Hyperlipidemia is considered as one of the major systemic factors associated with the development of diabetic retinopathy, and animal models have documented that its presence in a hyperglycemic environment exacerbates cytosolic ROS production (via activation of the Rac1–Nox2 axis) and mitochondrial damage. Hyperglycemia also accelerates Rac1 transcription via dynamic DNA methylation–hydroxymethylation of its promoter. In diabetes, ceramide metabolism in the retina is impaired and its accumulation is increased. Our aim was to investigate the effect of inhibition of the rate limiting enzyme of the de novo ceramide biosynthesis, serine palmitoyl-transferase (SPT), on Rac1 activation in diabetic retinopathy. Using human retinal endothelial cells, transfected with SPT-siRNA, and incubated in 20 mM d-glucose in the presence or absence of 50 µM palmitate (glucolipotoxic and glucotoxic, respectively), activities of Rac1 and Nox2, and ROS levels were quantified. For Rac1 transcriptional activation, 5 hydroxymethyl cytosine (5hmC) levels at its promoter were quantified. Key parameters were confirmed in retinal microvessels from streptozotocin-induced diabetic mice on a normal diet (type 1 diabetic model) or on a high-fat diet (45% kcal, type 2 diabetic model), injected intravitreally with SPT-siRNA. Compared to normal glucose, cells in high glucose, with or without palmitic acid, had increased Rac1–Nox2–ROS signaling, Rac1 transcripts and 5hmC levels at its promoter. Inhibition of SPT by SPT-siRNA or myriocin prevented glucotoxic- and glucolipotoxic-induced increase in Rac1–Nox2–ROS signaling and 5hmC at the Rac1 promoter. Similarly, in both type 1 and type 2 diabetic mouse models, SPT-siRNA attenuated the increase in the Rac1–Nox2–ROS axis and 5hmC at the Rac1 promoter. Thus, inhibition of the rate limiting enzyme of ceramide de novo biosynthesis, SPT, regulates activation of DNA methylation–hydroxymethylation machinery and prevents increased Rac1 transcription. This ameliorates the activation of Rac1–Nox2 signaling and protects the mitochondria from damaging cytosolic ROS, which prevents accelerated capillary cell loss. These results further raise the importance of regulating lipid levels in diabetic patients with dyslipidemia.

P.80 Respiratory chain dysfunction in dermatomyositis is associated with mitochondrial DNA depletion

Patients with dermatomyositis suffer from reduced aerobic metabolism contributing to impaired muscle function, which has been linked to cytochrome c oxidase (COX) deficiency in muscle tissue. This mitochondrial respiratory chain dysfunction is typically seen in perifascicular regions, which also show the most intense inflammatory reaction along with capillary loss and muscle fiber atrophy. The objective of this study was to investigate the pathobiology of the oxidative phosphorylation deficiency in dermatomyositis. Muscle biopsy specimens with perifascicular COX deficiency from five juveniles and seven adults with early untreated dermatomyositis were investigated. Immunohistochemical analyses of subunits in the respiratory chain were performed including complex I (subunit NDUFB8), complex II (succinate dehydrogenase, subunit SDHB) and complex IV (COX, subunit MTCO1). The results showed a profound deficiency of complex I and IV in the perifascicular regions with enzyme histochemical COX deficiency, whereas succinate dehydrogenase activity and complex II were preserved. In situ hybridization of mitochondrial RNA showed depletion of mitochondrial DNA (mtDNA) transcripts in the perifascicular regions. Analysis of mtDNA by next generation deep sequencing and quantitative PCR in affected muscle regions showed an overall reduction of mtDNA copy number particularly in the perifascicular regions. In conclusion, the respiratory chain dysfunction in dermatomyositis muscle is associated with mtDNA depletion causing deficiency of complex I and IV which are partially encoded by mtDNA, whereas complex II which is entirely nuclear gene encoded is preserved. The depletion of mtDNA indicates a perturbed replication of mtDNA partially explaining the muscle pathology and the disturbed aerobic metabolism.

The Relationship Between Gluten Enteropathy and Nail Capileroscopy Findings and Disease Activation

Objective: Nailfold Videocapilloroscopy (NVC) is an examination method that is used as an aid in the diagnosis, follow-up, and treatment strategy of rheumatic diseases such as systemic sclerosis, systemic lupus erythematosus, rheumatoid arthritis, and gives an idea about microcirculation by examining the vascular bed. It is a cheap, easily applicable, and quickly accessible method. Because of these features, we aimed to use the NVC method in patients with Gluten Enteropathy (GE) to determine whether this method will be a helpful technique in the diagnosis, activation decision, remission follow-up, and treatment strategy in patients with GE. Methods: In this study, 67 patients diagnosed with GE (n=35 disease-active group (AGE), n=32 disease-related remission group (RGE), and control group (CG)-27 healthy people whose diagnosis of GE was ruled out were included in this study. Group and CG were divided into ten parameters in capillary pathologies (capillary density loss, dilated capillary, giant capillary, microhemorrhage, avascular area, tortuosity, branched capillary, disorganization, extravasation, angiogenesis). They were divided into two groups as RGE and compared with the results obtained from NVC measurements Results: When patients diagnosed with GE and CG were evaluated in terms of capillary disorder with NVC, While all of the patients with capillary disorders were in the GE group, no capillary disorders were found in the control group (p&amp;lt;0.01). When patients diagnosed with GE were divided into two groups (AGE and RGE), NVC measurements were compared; All patients with capillary disorders were found in the AGE group (p&amp;lt;0.01). Capillary density loss and/or avascular area were detected in 80.9% of patients with capillary disorders. Conclucion: Our study found a statistically significant difference in NVC measurements between GE patients and CG (p&amp;lt;0.01). The fact that all patients with capillary disorders were in the active group in terms of the disease and no capillary disorders were detected in any patients in remission showed that this method could be used as an auxiliary technique in the diagnosis of GE, making the decision of activation or remission, monitoring the disease and determining treatment strategies.

The presence of sodium glucose co-transporter 2 in mesangial cells and pericytes and its roles in mesangial lesions and in capillaries under diabetic and ischemic conditions

SGLT2 is expressed in mesangial cells and pericytes, and is upregulated byhigh glucose and ischemia. Upregulated SGLT2 in both cells might directly worsen ischemia in kidney interstitial legion, heart and brain. The overexpression of SGLT2 in these cells could induce various organ failures via damages or loss of capillaries and dysfunctions of mesangial cells, which are attenuated by SGLT2 inhibitors.

P23 Antibody negative Systemic Sclerosis

Introduction/BackgroundThis case explores some classic rheumatology topics - systemic sclerosis and antibodies. It is now well recognised that there are a magnitude of different antibodies, beside the typical anti-Scl-70, anticentromere or anti-RNA polymerase III, that can be associated with systemic sclerosis however in 5-10% of cases all antibody tests are negative, yet the patient still displays characteristic clinical features of the disease. We need to ensure such patients are correctly diagnosed, followed up and managed.Description/MethodThis case looks at the uncommon presentation of systemic sclerosis without any positive antibody results. I consulted with a 53-year-old male with a past medical history of hypertension, type 2 diabetes, GORD, hypothyroidism, depression and hyperlipidaemia. He had previously been seen in the rheumatology clinic with costochondritis and Raynaud’s phenomenon. At this time, he had antibody tests which were all negative (ANA and ENA). He had nailfold capillaroscopy which showed some concerns with widening of capillaries in areas and some linear haemorrhages. This was felt to not be diagnostic of Systemic sclerosis but advised continued follow up and repeat tests. Unfortunately, he failed to attend a subsequent clinic appointment and was lost to follow up. He was referred back to the Rheumatology department in 2021 with worsening symptoms of Raynaud’s. At this time he also described symptoms of reflux and dysphagia. He was suffering from generalised fatigue and pains. On examination he had tightening of the skin on his hands and was prayer sign positive. His antibody tests were repeated, again all coming back negative. We then arranged for repeat nailfold capillaroscopy and thermographic testing. This showed very little rewarming after cold challenge with delay even at 30 degrees. The nailfold capillaroscopy showed generalised widening of the capillaries with occasional distortion and tortuosity. There were multiple linear haemorrhages. This study was reported showing definite abnormality in keeping with a diagnosis of systemic sclerosis.Discussion/ResultsThis highlights the rare but now well recognised phenomenon of antibody negative systemic sclerosis which makes up around 5-10% of cases. It also shows how nailfold capillaroscopy can be a key tool in diagnosing this, and the features to look out for in order to do so - capillary widening, haemorrhage and loss of architecture. The case demonstrates some of the more common features of antibody negative systemic sclerosis including a higher proportion of male patients and prominent GI features. There are generally less severe vascular features including pulmonary hypertension and telangiectasia, with Raynaud’s less often leading to digital ulcers and pitting. In this case we suggested uptitration of his nifedipine to try and help control his Raynaud’s symptoms. Given the significant GI features this patient was experiencing we referred him on for further evaluation by the gastroenterology team. I think some aspects of this case were blurred by his comorbidities, especially diabetes, and some of his skin disease and pains were attributed to this. However, with hindsight and seeing how his symptoms progressed, I think we could question if more could have been done earlier in the course of this patient’s disease - maybe when the initial abnormal, if not diagnostic, capillaroscopy results were available. Unfortunately, the patient not attending further clinic appointments reduced the opportunities to do this, but this also raised a good question about how we try to ensure patient engagement and deal with patients who don’t attend follow up.Key learning points/ConclusionThis was an excellent case for us to be able to learn about systemic sclerosis and the spectrum of disease it encompasses. It was interesting to learn about the different features and presentations of the disease, and how these different phenotypes correlate to certain antibodies classically being present, or as in this case, not present. It also enabled me to consider how we manage complex patients with multiple comorbidities and variable compliance. I feel that this conference would allow us to learn from other complex cases and how to best manage patients with similar issues, including systemic sclerosis, prominent pain issues, and gastrointestinal issues within rheumatology.

Capillary rarefaction during bed rest is proportionally less than fibre atrophy and loss of oxidative capacity.

Muscle disuse from bed rest or spaceflight results in losses in muscle mass, strength and oxidative capacity. Capillary rarefaction may contribute to muscle atrophy and the reduction in oxidative capacity during bed rest. Artificial gravity may attenuate the negative effects of long-term space missions or bed rest. The aim of the present study was to assess (1) the effects of bed rest on muscle fibre size, fibre type composition, capillarization and oxidative capacity in the vastus lateralis and soleus muscles after 6 and 55days of bed rest and (2) the effectiveness of artificial gravity in mitigating bed-rest-induced detriments to these parameters. Nineteen participants were assigned to a control group (control, n=6) or an intervention group undergoing 30min of centrifugation (n=13). All underwent 55days of head-down tilt bed rest. Vastus lateralis and soleus biopsies were taken at baseline and after 6 and 55days of bed rest. Fibre type composition, fibre cross-sectional area, capillarization indices and oxidative capacity were determined. After just 6days of bed rest, fibre atrophy (-23.2±12.4%, P<0.001) and reductions in capillary-to-fibre ratio (C:F; 1.97±0.57 vs. 1.56±0.41, P<0.001) were proportional in both muscles as reflected by a maintained capillary density. Fibre atrophy proceeded at a much slower rate between 6 and 55days of bed rest (-11.6±12.1% of 6days, P=0.032) and was accompanied by a 19.1% reduction in succinate dehydrogenase stain optical density (P<0.001), without any further significant decrements in C:F (1.56±0.41 vs. 1.49±0.37, P=0.459). Consequently, after 55days of bed rest, the capillary supply-oxidative capacity ratio of a fibre had increased by 41.9% (P<0.001), indicating a capillarization in relative excess of oxidative capacity. Even though the heterogeneity of capillary spacing (LogR SD) was increased after 55days by 12.7% (P=0.004), tissue oxygenation at maximal oxygen consumption of the fibres was improved after 55days bed rest. Daily centrifugation failed to blunt the bed-rest-induced reductions in fibre size and oxidative capacity and capillary rarefaction. The relationship between fibre size and oxidative capacity with the capillary supply of a fibre is uncoupled during prolonged bed rest as reflected by a rapid loss of muscle mass and capillaries, followed at later stages by a more than proportional loss of mitochondria without further capillary loss. The resulting excessive capillary supply of the muscle after prolonged bed rest is advantageous for the delivery of substrates needed for subsequent muscle recovery.

The VEGF Inhibitor Soluble Fms-like Tyrosine Kinase 1 Does Not Promote AKI-to-CKD Transition.

(1) Background: Soluble Fms-like tyrosine kinase 1 (sFLT1) is an endogenous VEGF inhibitor. sFLT1 has been described as an anti-inflammatory treatment for diabetic nephropathy and heart fibrosis. However, sFLT1 has also been related to peritubular capillary (PTC) loss, which promotes fibrogenesis. Here, we studied whether transfection with sFlt1 aggravates experimental AKI-to-CKD transition and whether sFLT1 is increased in human kidney fibrosis. (2) Methods: Mice were transfected via electroporation with sFlt1. After confirming transfection efficacy, mice underwent unilateral ischemia/reperfusion injury (IRI) and were sacrificed 28 days later. Kidney histology and RNA were analyzed to study renal fibrosis, PTC damage and inflammation. Renal sFLT1 mRNA expression was measured in CKD biopsies and control kidney tissue. (3) Results: sFlt1 transfection did not aggravate renal fibrosis, PTC loss or macrophage recruitment in IRI mice. In contrast, higher transfection efficiency was correlated with reduced expression of pro-fibrotic and pro-inflammatory markers. In the human samples, sFLT1 mRNA levels were similar in CKD and control kidneys and were not correlated with interstitial fibrosis or PTC loss. (4) Conclusion: As we previously found that sFLT1 has therapeutic potential in diabetic nephropathy, our findings indicate that sFLT1 can be administered at a dose that is therapeutically effective in reducing inflammation, without promoting maladaptive kidney damage.

Therapy experience in alopecia after COVID-19

The coronavirus infection (COVID-2019) caused by SARS-CoV-2 continues to be studied, the virus mutates, new strains differing from the previous ones by rapid spread appear. Due to the difficult epidemiological situation around the world, the long-term course of the pandemic, doctors of various disciplines are increasingly facing its consequences. 25% of patients have various types of alopecia following infection. Suppression of some growth factors due to the collapse of the immune privilege, local and systemic inflammation, oxidative stress, tissue hypoxia, microvascular insufficiency manifested by persistent capillary spasm influence hair loss. The article presents data from the analysis, out-patient medical records of patients aged 19 to 60 years who sought advice from the Ekaterininskaya clinic network with complaints of hair loss and past history of confirmed coronavirus infection. Due to the variety of trigger factors and multifaceted pathogenetic mechanisms of influence on the hair follicle in coronavirus infection, it is more appropriate to use drugs with a complex mechanism of action for the treatment. The patients received a complex drug for the treatment of alopecia, 1 tablet three times a day for 2 months, and a peptide lotion for cutaneous use with active ingredients, which activate cellular metabolism in hair follicles, encourage the hair follicles to switch to the active growth phase. A positive result during treatment was achieved in 92% of patients. Good tolerability, absence of side effects allow us to administer the drugs we use in a range of therapeutic measures in patients with diffuse hair thinning following coronavirus infection, both in monotherapy and in combination with other drugs.

WIDEFIELD SWEPT-SOURCE OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY FINDINGS IN WAGNER SYNDROME.

To describe novel clinical and angiographic findings in Wagner syndrome. A retrospective case series of three related patients with Wagner syndrome. Patients underwent standard optical coherence tomography (OCT), B-scan ultrasonography, and fluorescein angiography in addition to wide field swept-source OCT angiography (WF SS-OCTA) (PLEX Elite 9000, Carl Zeiss Meditec Inc). Patients underwent genetic testing for a panel of hereditary vitreoretinopathies. Three related patients with Wagner syndrome were identified. All were found to have prominent vitreous strands, abnormal vitreoretinal adhesions, peripheral retinal holes, and varying degrees of myopia. A mid-peripheral tractional ridge was identified in all six eyes. All patients were positive for a known pathologic intron variant in the VCAN gene (4004-5T-A). Wide field swept-source OCT angiography (12 mm × 12 mm) was performed in two patients and demonstrated perivascular capillary loss in the superficial capillary plexus along the arcades bilaterally. One patient demonstrated associated retinal atrophy within the area of capillary loss. The capillary loss extended beyond the margin of retinal atrophy. The unusual finding of a mid-peripheral tractional ridge of the retina associated with myopia led to a genetic diagnosis of Wagner syndrome. Widefield swept-source OCT angiography demonstrated a novel feature of perivascular loss of the superficial retinal capillary plexus. This result suggests that vitreous traction may cause localized microvasculature dysfunction and subsequent retinal atrophy in Wagner syndrome. This is the first known evaluation of Wagner syndrome using OCT angiography.

Respiratory chain dysfunction in perifascicular muscle fibres in patients with dermatomyositis is associated with mitochondrial DNA depletion.

Patients with dermatomyositis (DM) suffer from reduced aerobic metabolism contributing to impaired muscle function, which has been linked to cytochrome c oxidase (COX) deficiency in muscle tissue. This mitochondrial respiratory chain dysfunction is typically seen in perifascicular regions, which also show the most intense inflammatory reaction along with capillary loss and muscle fibre atrophy. The objective of this study was to investigate the pathobiology of the oxidative phosphorylation deficiency in DM. Muscle biopsy specimens with perifascicular COX deficiency from five juveniles and seven adults with DM were investigated. We combined immunohistochemical analyses of subunits in the respiratory chain including complex I (subunit NDUFB8), complex II (succinate dehydrogenase, subunit SDHB) and complex IV (COX, subunit MTCO1) with in situ hybridisation, next generation deep sequencing and quantitative polymerase chain reaction (PCR). There was a profound deficiency of complexes I and IV in the perifascicular regions with enzyme histochemical COX deficiency, whereas succinate dehydrogenase activity and complex II were preserved. In situ hybridisation of mitochondrial RNA showed depletion of mitochondrial DNA (mtDNA) transcripts in the perifascicular regions. Analysis of mtDNA by next generation deep sequencing and quantitative PCR in affected muscle regions showed an overall reduction of mtDNA copy number particularly in the perifascicular regions. The respiratory chain dysfunction in DM muscle is associated with mtDNA depletion causing deficiency of complexes I and IV, which are partially encoded by mtDNA, whereas complex II, which is entirely encoded by nuclear DNA, is preserved. The depletion of mtDNA indicates a perturbed replication of mtDNA explaining the muscle pathology and the disturbed aerobic metabolism.

Swept-source Optical Coherence Tomography Angiography in Active and Scarred Toxoplasmic Retinochoroiditis

ABSTRACT Purpose To describe optical coherence tomography angiography (OCTA) findings in eyes with active and scarred toxoplasmic retinochoroiditis. Methods OCTA scans in active (25 eyes) and scarred (17 eyes) retinochoroiditis were retrospectively reviewed. Results In active lesions, OCTA findings included a non-detectable flow signal area in retinal vascular plexuses and choriocapillaris in all 25 eyes (100%), an abnormal intraretinal vascular process in 2 eyes (8%), and an associated area of retinal flow deficit secondary to branch retinal artery occlusion in one eye (4%). In scarred lesions, OCTA findings included a flow deficit area in retinal vascular plexuses and choriocapillaris in all 17 eyes (100%), a visibility of larger deeper choroidal vessels at the level of choriocapillaris in 9 eyes (53%), and a well-defined intraretinal vascular network in one eye (5.9%). Peripapillary scars were associated on OCTA with wedge-shaped loss of radial peripapillary capillaries with corresponding localized retinal nerve fiber layer defect and visual field loss. Conclusion OCTA allows to non-invasively detect retinal and choroidal vascular changes in active and scarred toxoplasmic retinochoroiditis.

Pro-Angiogenetic Effects of Purified Extracts from Helix aspersa during Zebrafish Development

Helix aspersa is a species of land snail belonging to the Helicidae family, widespread in the Mediterranean and continental area up to Northern Europe. In some areas it is appreciated as a food, but is mostly considered a parasite of gardens and cultivated fields. The mucus of Helix aspersa has found multiple applications in the cosmetic and health fields. In the present study, we investigated for the first time the angiogenetic properties of purified extracts from Helix aspersa using a transgenic zebrafish line Tg (kdrl:EGFP). The angiogenesis induced by purified snail extracts was demonstrated by their capability to increase the three well-established parameters of angiogenesis: generation of intersegmental vessels, modeling of caudal venous plexus, and formation of sub-intestinal venous plexus. The effects appeared to be mediated by the vascular endothelial growth factor (VEGF) pathway, being prevented by pretreatment of embryos with the selective VEGF receptor antagonist SU5416, and supported by the increased VEGF mRNA levels found in snail-extract-treated embryos. Insufficient vascular supply is underlined by low VEGF signaling, primarily because of its indispensable role in preventing capillary loss. Our findings might have a pharmacological impact by counteracting VEGF hypofunction and promoting angiogenesis to maintain adequate microvascular and vascular density in normal and suffering tissues and organs.

Examination of nailfold videocapillaroscopy findings in ANCA-associated vasculitis.

The objective of this study was to evaluate nailfold videocapillaroscopy (NVC) as a useful tool for assessing the disease activity of ANCA-associated vasculitis (AAV). This study enrolled 51 patients with AAV and 21 healthy controls. We scored NVC findings semiquantitatively, and compared them between AAV patients and controls. We examined the association of NVC findings with disease activity indicators, histopathological findings of skin biopsies, and high-resolution CT (HRCT) scores in AAV. Additionally, we repeatedly rated the NVC findings 3 months after immunosuppressive therapy. Of the 51 enrolled patients, 36 (70.6%) showed a microangiopathy pattern and 4 (7.8%) showed a scleroderma pattern in AAV. The scores for microhaemorrhage, capillary loss, neoangiogenesis, and tortuosity were significantly higher in the AAV group than in the control group. NVC abnormalities correlated with the severity of skin, lung and kidney involvement. The scores of giant capillaries significantly correlated with the total BVAS and the chest BVAS; the scores of capillary loss correlated with the chest BVAS and the renal BVAS. The scores of microhaemorrhage significantly correlated with perivascular inflammatory cell infiltrations in the upper dermis of the purpura and tended to correlate with the total ground-glass opacity and consolidation scores on HRCT. In addition, capillary loss scores had a significant positive correlation with serum creatinine levels. Additionally, the microhaemorrhage scores were significantly reduced after 3 months of immunosuppressive therapy. In AAV patients, NVC abnormalities are significantly associated with disease severity. This result suggests that NVC is a useful tool for assessing the disease activity and treatment response in AAV.