Granulomatosis with Polyangiitis Discovered Because of Repeated Upper Eyelid Swelling.

Background and objectives: The initial symptom that triggers granulomatosis with polyangiitis (GPA) diagnosis is rarely ocular. We describe a case with a single ocular lesion identified as probable GPA due to proteinase 3 (PR3)-antineutrophil cytoplasmic antibody (ANCA)-positivity according to the diagnostic criteria of the Ministry of Health in Japan; the lesion repeatedly worsened. Materials and methods: A 25-year-old female visited the Department of Ophthalmology, Asahi General Hospital, with upper eyelid swelling and conjunctival and episcleral hyperemia of the left eye. Both hordeolum and eyelid cellulitis were suspected, as the condition was resistant to treatment with antibiotic eye drops. Episcleritis was suspected due to localized hyperemia in the upper part of the eye. Upon treatment with antibacterial agents and steroid eye drops, the swelling and the hyperemia repeatedly worsened every week. Results: Blood samples were positive for PR3-ANCA, and GPA with an isolated ocular lesion was considered. After oral steroid treatment, the patient had no recurrence for 4 years. There was no systemic involvement in the upper respiratory tract, lungs, or kidneys. Conclusions: Diagnosing GPA with ocular symptoms as initial manifestations is challenging. GPA should be considered in treatment-resistant eyelid, orbital, and episcleral lesions, even at a young age.

Background factors determining the time to intraocular lens dislocation.

PURPOSE: To clarify the characteristics of intraocular lens (IOL) dislocation requiring IOL suture or intraocular scleral fixation. METHODS: This retrospective consecutive case series included 21 eyes (21 patients) who required sutured or sutureless intrascleral IOL fixation following IOL extraction owing to IOL dislocation at the outpatient clinic in the Department of Ophthalmology, Saitama Red Cross Hospital, Japan, between January and December 2019. Medical records were retrospectively reviewed for background diseases, location of the dislocated IOL (intracapsular/extracapsular), insertion of a capsular tension ring (CTR), and the period from IOL insertion to dislocation. RESULTS: We included 21 eyes of 21 patients who required IOL suture or intrascleral fixation for IOL dislocation at our clinic from January to December 2019 were included. The most common background disease was pseudoexfoliation syndrome (four cases), followed by atopic dermatitis, dysplasia/dehiscence of the zonule, post-retinal detachment surgery, high myopia, and uveitis (three cases each). At the time of dislocation, the IOLs were either intracapsular (16 cases, including 3 cases with CTR insertion) or extracapsular (5 cases). The time from IOL insertion to IOL dislocation was 13.7 ± 8.1 years (maximum: 31.3 years, minimum: 1.7 years). CONCLUSIONS: In this study, all 21 cases represented late IOL dislocations occurring after 3 months postoperatively. Among these late IOL dislocation cases, IOL dislocation occurred in a short-medium period of time, especially in those with CTR insertion and weakness/dehiscence of the zonule, with an average of 3 to 5 years postoperatively. We propose referring to these cases as intermediate-term IOL dislocation.

Loss of Down syndrome critical region-1 leads to cholesterol metabolic dysfunction that exaggerates hypercholesterolemia in ApoE-null background.

Down syndrome critical region (DSCR)-1 functions as a feedback modulator for calcineurin-nuclear factor for activated T cell (NFAT) signals, which are crucial for cell proliferation and inflammation. Stable expression of DSCR-1 inhibits pathological angiogenesis and septic inflammation. DSCR-1 also plays a critical role in vascular wall remodeling associated with aneurysm development that occurs primarily in smooth muscle cells. Besides, Dscr-1 deficiency promotes the M1-to M2-like phenotypic switch in macrophages, which correlates to the reduction of denatured cholesterol uptakes. However, the distinct roles of DSCR-1 in cholesterol and lipid metabolism are not well understood. Here, we show that loss of apolipoprotein (Apo) E in mice with chronic hypercholesterolemia induced Dscr-1 expression in the liver and aortic atheroma. In Dscr-1-null mice fed a high-fat diet, oxidative- and endoplasmic reticulum (ER) stress was induced, and sterol regulatory element-binding protein (SREBP) 2 production in hepatocytes was stimulated. This exaggerated ApoE-/--mediated nonalcoholic fatty liver disease (NAFLD) and subsequent hypercholesterolemia. Genome-wide screening revealed that loss of both ApoE and Dscr-1 resulted in the induction of immune- and leukocyte activation-related genes in the liver compared with ApoE deficiency alone. However, expressions of inflammation-activated markers and levels of monocyte adhesion were suspended upon induction of the Dscr-1 null background in the aortic endothelium. Collectively, our study shows that the combined loss of Dscr-1 and ApoE causes metabolic dysfunction in the liver but reduces atherosclerotic plaques, thereby leading to a dramatic increase in serum cholesterol and the formation of sporadic vasculopathy.

Massive intraocular hemorrhage, presumably from the central retinal artery after cataract surgery, and difficult hemostasis during vitrectomy: a case report.

BACKGROUND: Severe intraocular hemorrhage is a rare complication of cataract surgery due to the recent generalization of minimal-incision cataract surgery. We report a case of a massive intraocular hemorrhage that probably originated from the central retinal artery after cataract surgery, in which hemostasis was difficult to achieve during vitrectomy. CASE PRESENTATION: An 86-year-old woman was referred to our department for intraocular lens (IOL) dislocation after undergoing cataract surgery. Massive intraocular hemorrhage was observed during the initial visit to our department. She underwent pars plana vitrectomy (PPV) and IOL repositioning under local anesthesia. However, the hemorrhage could not be removed completely because of continued massive intraoperative bleeding from the posterior fundus, and it was extremely difficult to achieve hemostasis during the initial surgery. At 7 days after the initial surgery, PPVs were performed under general anesthesia. Bleeding significantly decreased in the second surgery compared to the first. The bleeding probably originated from the central retinal artery on the optic disc; hemostasis was obtained by coagulation of the bleeding site with intraocular diathermy. After the second surgery, there was no exacerbation of bleeding and the patient's condition was stable. However, the patient's visual acuity showed no light perception after the second surgery. CONCLUSIONS: Massive intraocular hemorrhage may occur from the central retinal artery after undergoing cataract surgery. In such cases, surgery with general anesthesia with a lower maintained blood pressure (instead of surgery under local anesthesia) should be recommended, considering the possibility of difficult hemostasis in the event of bleeding from the retinal artery.

Loss of Down Syndrome Critical Region-1 Mediated-Hypercholesterolemia Accelerates Corneal Opacity Via Pathological Neovessel Formation.

OBJECTIVE: The calcineurin-NFAT (nuclear factor for activated T cells)-DSCR (Down syndrome critical region)-1 pathway plays a crucial role as the downstream effector of VEGF (vascular endothelial growth factor)-mediated tumor angiogenesis in endothelial cells. A role for DSCR-1 in different organ microenvironment such as the cornea and its role in ocular diseases is not well understood. Corneal changes can be indicators of various disease states and are easily detected through ocular examinations. Approach and Results: The presentation of a corneal arcus or a corneal opacity due to lipid deposition in the cornea often indicates hyperlipidemia and in most cases, hypercholesterolemia. Although the loss of Apo (apolipoprotein) E has been well characterized and is known to lead to elevated serum cholesterol levels, there are few corneal changes observed in ApoE-/- mice. In this study, we show that the combined loss of ApoE and DSCR-1 leads to a dramatic increase in serum cholesterol levels and severe corneal opacity with complete penetrance. The cornea is normally maintained in an avascular state; however, loss of Dscr-1 is sufficient to induce hyper-inflammatory and -oxidative condition, increased corneal neovascularization, and lymphangiogenesis. Furthermore, immunohistological analysis and genome-wide screening revealed that loss of Dscr-1 in mice triggers increased immune cell infiltration and upregulation of SDF (stromal derived factor)-1 and its receptor, CXCR4 (C-X-C motif chemokine ligand receptor-4), potentiating this signaling axis in the cornea, thereby contributing to pathological corneal angiogenesis and opacity. CONCLUSIONS: This study is the first demonstration of the critical role for the endogenous inhibitor of calcineurin, DSCR-1, and pathological corneal angiogenesis in hypercholesterolemia induced corneal opacity.

Diagnosis of cytomegalovirus corneal endotheliitis using surgically removed Descemet's membrane and endothelium despite negative results with aqueous humor PCR: a case report.

BACKGROUND: Cytomegalovirus (CMV) has been known to cause unilateral corneal endotheliitis with keratic precipitates and localized corneal edema, iridocyclitis, and secondary glaucoma. CMV endotheliitis is diagnosed based on clinical manifestations and viral examination using qualitative polymerase chain reaction (PCR) of the aqueous humor. CASE PRESENTATION: An 80-year-old woman was referred to our department for bullous keratopathy. Pigmented keratic precipitates were found in the right eye without significant anterior chamber inflammation. After 8 months there was inflammation relapse with mutton fat keratic precipitates and PCR on aqueous humor was performed, with negative results for CMV, herpes simplex virus, and varicella zoster virus. Keratic precipitates disappeared with steroid instillation, and Descemet-stripping automated endothelial keratoplasty (DSAEK) was performed for the right eye. CMV-DNA was positive at 6.0 × 102 copies/ GAPDH 105 copies in real time PCR of corneal endothelial specimen removed during DSAEK with negative results for all the other human herpes viruses. After diagnosis of CMV corneal endotheliitis, treatment with systemic and topical ganciclovir was initiated and there was resolution of symptoms. No recurrence of iridocyclitis or corneal endotheliitis was observed at 6 months follow up. CONCLUSIONS: This case report suggests that PCR should be performed using the endothelium removed during DSAEK for bullous keratopathy of an unknown cause, even if PCR for aqueous humor yields negative results.

Oxidative stress induces ferroptotic cell death in retinal pigment epithelial cells.

The dysfunction and cell death of retinal pigment epithelial (RPE) cells are hallmarks of late-stage dry (atrophic) age-related macular degeneration (AMD), for which no effective therapy has yet been developed. Previous studies have indicated that iron accumulation is a source of excess free radical production in RPE, and age-dependent iron accumulation in RPE is accelerated in patients with dry AMD. Although the pathogenic role of oxidative stress in RPE in the development of dry AMD is widely accepted, the mechanisms of oxidative stress-induced RPE cell death remain elusive. Here, we show that ferroptotic cell death, a mode of regulated necrosis mediated by iron and lipid peroxidation, is implicated in oxidative stress-induced RPE cell death in vitro. In ARPE-19 cells we observed that the ferroptosis inhibitors ferrostatin-1 and deferoxamine (DFO) rescued tert-butyl hydroperoxide (tBH)-induced RPE cell death more effectively than inhibitors of apoptosis or necroptosis. tBH-induced RPE cell death was accompanied by the three characteristics of ferroptotic cell death: lipid peroxidation, glutathione depletion, and ferrous iron accumulation, which were all significantly attenuated by ferrostatin-1 and DFO. Exogenous iron overload enhanced tBH-induced RPE cell death, but this effect was also attenuated by ferrostatin-1 and DFO. Furthermore, mRNA levels of numerous genes known to regulate iron metabolism were observed to be influenced by oxidative stress. Taken together, our observations suggest that multiple modes of cell death are involved in oxidative stress-induced RPE cell death, with ferroptosis playing a particularly important role.

Poor intraoperative visibility and postoperative astigmatism associated with trabecular micro-bypass stent for corneal dystrophy: A case report.

RATIONALE: There are reports of safe cataract surgery in eyes with posterior polymorphous corneal dystrophy (PPCD); however, to our knowledge, there are no reports of minimally invasive glaucoma surgery (MIGS) in eyes with PPCD. Herein, we report a case of poor intraoperative visibility with gonioscopy, postoperative corneal edema, and corneal astigmatism in eyes with PPCD treated with trabecular micro-bypass stent combined with cataract surgery. PATIENT CONCERNS/DIAGNOSIS: A 78-year-old man was referred to our hospital for MIGS. He presented with bilateral corneal endothelial vesicular changes and band lesions. Endothelial cell density was 2983/2871 cells/mm2 (right/left eye), central corneal thickness was 581 µm/572 µm, best-corrected visual acuity values (Snellen equivalent) were 20/32 (right) and 20/100 (left), and corneal astigmatism was -2.7D in the right eye and -2.5D in the left eye. INTERVENSIONS/ OUTCOMS: After phacoemulsification and aspiration with intraocular lens implantation with a 2.4-mm corneal incision in both eyes, trabecular micro-bypass stents were inserted successfully despite the poor intraoperative visibility with gonioscopy. One week after surgery, the central corneal thickness was 614 µm/609 µm, and Descemet's membrane folds and mild corneal edema were observed. Best-corrected decimal visual acuity was 20/40 for the right eye and 20/50 for the left eye. In the left eye, total corneal astigmatism increased from -2.5D to -5.5D. Corneal astigmatism and edema showed gradual improvement. LESSONS: Although reports have shown that cataract surgery can be safely performed in eyes with PPCD, MIGS in eyes with PPCD may require caution regarding intraoperative visibility with gonioscopy and visual function in the early postoperative period.

Effects of reduction in intraocular pressure after trabeculotomy on axial length and intraocular lens selection.

PURPOSE: To investigate the relationship between intraocular pressure (IOP) and axial length (AL) and to compare the refractive predicted error in patients who have undergone cataract surgery alone or in combination with trabeculotomy. SETTING: Hospital. DESIGN: Single-center, retrospective, case-control. METHODS: The medical records of patients who had undergone cataract surgery alone or in combination with trabeculotomy using the Tanito microhook were retrospectively reviewed. Patients were grouped into cataract surgery alone (CAT) or cataract surgery combined with trabeculotomy (LOT) groups. Demographic data, preoperative and postoperative IOP and AL, and surgically induced astigmatism (SIA) were analyzed before and 1 month postoperatively to evaluate the interplay between IOP, AL, and refractive outcomes. RESULTS: 52 eyes (52 patients) underwent LOT, and 67 eyes (67 patients) underwent CAT. The mean IOP at baseline did not differ between the groups; the change in IOP (dIOP) was significantly higher in the LOT group than in the CAT group. The mean AL at baseline and the change in AL (dAL) were 24.0 ± 1.2 mm and 0.16 ± 0.11 mm, respectively, in the LOT group, and 23.8 ± 1.1 mm and 0.11 ± 0.070 mm, respectively, in the CAT group. The difference in dAL was also significant. In the LOT group, dIOP and dAL were significantly correlated. The mean SIA vectors did not significantly differ between the groups. CONCLUSIONS: AL decreased because of the reduction in IOP after cataract surgery combined with trabeculotomy. Consequently, the refractive target error was greater, and the postoperative refractive outcome showed a tendency toward hyperopia.

Clinical Outcomes of Phacoemulsification in Japanese Patients Receiving First and Revised Second-Generation Trabecular Microbypass Stents.

PURPOSE: This study evaluated the effectiveness and safety of first and revised second-generation trabecular microbypass stent insertion [iStent (IS) and iStent inject W (IW)] in cataract surgery. DESIGN: Single-center, retrospective, cohort study. METHODS: The study included 176 eyes that received trabecular microbypass stents combined with cataract surgery at the Saitama Red Cross Hospital between September 2017 and September 2021. Patients were divided into IS and IW groups depending on the implant type. Demographic characteristics, intraocular pressure (IOP), and the number of antiglaucoma medications (Med) were analyzed preoperatively and 12 months postoperatively. In addition, postoperative complications were compared between the groups. RESULTS: IS and IW were implanted in 86 eyes and 90 eyes, respectively. IOP and Med at 1, 3, 6, 9, and 12 months decreased significantly from baseline in both groups ( P = 0.04, P < 0.001, P < 0.001, P < 0.001, and P < 0.001, respectively, for IOP in the IS group; P = 0.02, P < 0.001, P < 0.001, P < 0.001, and P < 0.001, respectively, for IOP in the IW group; P = 0.03, P = 0.002, P < 0.001, P < 0.001, and P < 0.001, respectively, for Med in the IS group; and P < 0.001 for all time points for Med in the IW group). IOP did not vary significantly between the groups at 1, 3, 6, 9, and 12 months postoperatively. Med was significantly lower in IW than IS at 1, 3, 6, 9, and 12 months postoperatively ( P < 0.001, P < 0.001, P = 0.002, P = 0.002, and P = 0.002, respectively). Hyphema and IOP >30 mm Hg (spike) occurred in 1.2% and 4.4%, and 1.2% and 3.3% of patients in the IS and IW groups, respectively. The probability of successful discontinuation of medications at 12 months postoperatively was 10.5% and 41.1%, respectively ( P < 0.001). CONCLUSIONS: Postoperative Med was significantly lower in the IW group. Simultaneous insertion of IW in patients with glaucoma requiring cataract surgery may be preferred to IS because it reduces the burden of Med.

Synergistic effects of high hydrostatic pressure, mild heating, and amino acids on germination and inactivation of Clostridium sporogenes spores.

The synergistic effects of high hydrostatic pressure (HHP), mild heating, and amino acids on the germination of Clostridium sporogenes spores were examined by determining the number of surviving spores that returned to vegetative growth after pasteurization following these treatments. Pressurization at 200 MPa at a temperature higher than 40°C and treatment with some of the 19 l-amino acids at 10 mM or higher synergistically facilitated germination. When one of these factors was omitted, the level of germination was insignificant. Pressures of 100 and 400 MPa were less effective than 200 MPa. The spores were effectively inactivated by between 1.8 and 4.8 logs by pasteurization at 80°C after pressurization at 200 MPa at 45°C for 120 min with one of the amino acids with moderate hydrophobicity, such as Leu, Phe, Cys Met, Ala, Gly, or Ser. However, other amino acids showed poor inactivation effects of less than 0.9 logs. Spores in solutions containing 80 mM of either Leu, Phe, Cys, Met, Ala, Gly, or Ser were successfully inactivated by pasteurization by more than 5.4 logs after pressurization at 200 MPa at 70°C for 15 to 120 min. Ala and Met reduced the spore viability by 2.8 and 1.8 logs, respectively, by pasteurization at a concentration of 1 mM under 200 MPa at 70°C. These results indicate that germination of the spores is facilitated by a combination of high hydrostatic pressure, mild heating, and amino acids.

Anterior Chamber Shallowing from the Early Stage of Surgery and Suprachoroidal Effusion during Clear Corneal Small-Incision Cataract Surgery: A Case Report.

Suprachoroidal effusion (SCE) is a rarely observed complication due to the recent generalization of clear corneal small-incision cataract surgery. We report a case of anterior chamber shallowing (ACS) from the early stage of surgery and SCE during clear corneal small-incision cataract surgery. A 69-year-old man was referred to our department for primary open-angle glaucoma and grade 2 nuclear cataract. The intraocular pressure (IOP) was 18 and 12 mm Hg in the right and left eyes with the instillation of three antiglaucoma eye drops in both eyes, respectively, and deep anterior chamber and normal axial length were observed. At the age of 70 years, which was 4 months after the initial visit to our department, the IOP of the right eye increased to 30 mm Hg. Hence, cataract surgery and microhook ab interno trabeculotomy (µLOT) of the right eye were scheduled. Mild ACS was observed during continuous curvilinear capsulorhexis (CCC), and ACS worsened as the surgery progressed, making the surgery progressively challenging. SCE was observed by fundus examination after phacoemulsification and cortex removal, and the wound was immediately closed with a suture. The IOP was 28 mm Hg on postoperative day (POD) 1 and decreased to 14 mm Hg on POD 5. SCE disappeared on POD 12. On POD 18, intraocular lens implantation into the bag and µLOT were performed under general anesthesia. Subsequently, the IOP decreased to 15 mm Hg 3 months after the surgery. Mild ACS was already present at the time of CCC, so it is possible that SCE occurred in the early stage of surgery. If ACS is observed intraoperatively, especially if there are SCE risk factors, such as hypertension, glaucoma, and lung cancer, as in this case, and even if the eye has deep anterior chamber and normal axial length preoperatively, fundoscopic examination should be performed even at an early stage of clear corneal small-incision cataract surgery to rule out SCE.

Differential gene expression analysis using RNA sequencing: retinal pigment epithelial cells after exposure to continuous-wave and subthreshold micropulse laser.

PURPOSE: Subthreshold micropulse laser (SMPL) is more clinically efficient for the treatment of diabetic macular edema (DME) than the conventional continuous-wave (CW) laser. We aimed to characterize transcriptome changes after the application of these lasers and to compare the transcripts. METHODS: Human pluripotent stem cell-derived retinal pigment epithelial cells were exposed to laser irradiation. Differentially expressed genes (DEGs), distribution of heat shock protein (Hsp) family, gene expression profile, and gene ontology (GO) enrichment analysis based on RNA sequencing data were investigated at 3 h and 24 h after irradiation. RESULTS: CW laser induced more DEGs than SMPL (1771 vs. 520 genes). The expression of the Hsp family was confirmed in both groups: however, the induction patterns was different for different genes. GO enrichment analysis revealed that CW laser upregulated the expression of DEGs involved in vasculature development (GO: 0001944), related to apoptosis and repair after cell injury whereas SMPL upregulated the expression of DEGs involved in photoreceptor cell maintenance (GO: 0045494), photoreceptor cell development (GO: 0042461), and sensory perception of light stimuli (GO: 0050953). CONCLUSIONS: The results provide insights into the genetic responses and may contribute to the understanding of the molecular mechanisms of laser-induced thermal effects.

PITX1 inhibits the growth and proliferation of melanoma cells through regulation of SOX family genes.

Melanoma is one of the most aggressive types of cancer wherein resistance to treatment prevails. Therefore, it is important to discover novel molecular targets of melanoma progression as potential treatments. Here we show that paired-like homeodomain transcription factor 1 (PITX1) plays a crucial role in the inhibition of melanoma progression through regulation of SRY-box transcription factors (SOX) gene family mRNA transcription. Overexpression of PITX1 in melanoma cell lines resulted in a reduction in cell proliferation and an increase in apoptosis. Additionally, analysis of protein levels revealed an antagonistic cross-regulation between SOX9 and SOX10. Interestingly, PITX1 binds to the SOX9 promoter region as a positive regulatory transcription factor; PITX1 mRNA expression levels were positively correlated with SOX9 expression, and negatively correlated with SOX10 expression in melanoma tissues. Furthermore, transcription of the long noncoding RNA (lncRNA), survival-associated mitochondrial melanoma-specific oncogenic noncoding RNA (SAMMSON), was decreased in PITX1-overexpressing cells. Taken together, the findings in this study indicate that PITX1 may act as a negative regulatory factor in the development and progression of melanoma via direct targeting of the SOX signaling.

[Relationship between initial diagnosis and long-term prognosis in lupus nephritis].

UNLABELLED: To clarify the outcomes of patients with lupus glomerulonephritis (LGN), we performed a retrospective study of 31 patients (27 females and 4 males) with LGN between January 1975 and June 2006. All these 31 patients fulfilled the SLE criteria of the American College of Rheumatology evaluated by pathohistological diagnosis using renal biopsies. According to 2003 ISN/RPS classification, we reclassified all initial renal biopsies as class II 16%, class III 16%, class IV 48%, and class V 19.5%. Activity and chronicity indices were also calculated according to the scores proposed by Austin et al. All patients were treated by oral corticosteroids in induction therapy, then subsequeatly 18 patients (61%) were treated with intravenous methylprednisolone pulse therapy, and 16 patients with immunosuppressive agents (58%). Clinical remission rate was 94% by induction therapy and 13% by recurrence rate. Patient survival rate was 85% at 10 years and 76% at 20 years. Renal survival rate was 96% at 10 years and 86% at 20 years, 100% at 10 years and 80% at 20 years in ClassIV-G. In the multivariate Cox hazard analysis of the clinicopathologic factors, serum creatinine was selected as the most significant risk factor for death and/or end-stage renal failure (p=0.036). In addition, the chronicity index was also a significant risk factor for renal survival of LGN. CONCLUSION: This retrospective analysis of LGN showed better outcomes than expected. Overall, early diagnosis and suitable initial therapy may improve the renal survival of LGN in both groups of patients.

A high-salt diet enhances leukocyte adhesion in association with kidney injury in young Dahl salt-sensitive rats.

Salt-sensitive hypertension is associated with severe organ damage. Generating oxygen radicals is an integral component of salt-induced kidney damage, and activated leukocytes are important in oxygen radical biosynthesis. We hypothesized that a high-salt diet causes the upregulation of immune-related mechanisms, thereby contributing to the susceptibility of Dahl salt-sensitive rats to hypertensive kidney damage. For verifying the hypothesis, we investigated leukocytes adhering to retinal vessels when Dahl salt-sensitive rats were challenged with a high-salt (8% NaCl) diet using acridine orange fluoroscopy and a scanning laser ophthalmoscope. The high-salt diet increased leukocyte adhesion after 3 days and was associated with a significant increase in mRNA biosynthesis of monocyte chemotactic protein-1 and intercellular adhesion molecule-1 (ICAM-1) -related molecules in the kidney. Losartan treatment did not affect increased leukocyte adhesion during the early, pre-hypertensive phase of high salt loading; however, losartan attenuated the adhesion of leukocytes during the hypertensive stage. Moreover, the inhibition of leukocyte adhesion in the pre-hypertensive stage by anti-CD18 antibodies decreased tethering of leukocytes and was associated with the attenuation of functional and morphological kidney damage without affecting blood pressure elevation. In conclusion, a high-salt challenge rapidly increased leukocyte adhesion through the over-expression of ICAM-1. Increased leukocyte adhesion in the pre-hypertensive stage is responsible for subsequent kidney damage in Dahl salt-sensitive rats. Immune system involvement may be a key component that initiates kidney damage in a genetic model of salt-induced hypertension.

Erratum: A high-salt diet enhances leukocyte adhesion in association with kidney injury in young Dahl salt-sensitive rats.

This corrects the article DOI: 10.1038/hr.2017.31.

The angiotensin II receptor antagonist, losartan, enhances regulator of G protein signaling 2 mRNA expression in vascular smooth muscle cells of Wistar rats.

Angiotensin II (Ang II) reportedly enhances regulator of G-protein signaling 2 (RGS2), thus making a negative feedback loop for Ang II signal transduction. However, few studies have reported whether Ang II receptor (ATR) antagonists influence RGS2 mRNA expression. We investigated RGS2 mRNA expression when Ang II binding to ATR was blocked with Ang II subtype-1 receptor (AT1R) blockers using vascular smooth muscle cells from the thoracic aorta of male Wistar rats. RGS2 mRNA expression significantly increased with Ang II stimulation, and this increase was almost completely abolished by olmesartan, a potent AT1R-specific blocker. Ang II subtype-2 receptor (AT2R) was not involved in Ang II-mediated RGS expression. In contrast, the AT1R blocker, losartan, partially decreased Ang II-mediated RGS2 mRNA expression because this antagonist directly stimulated RGS2 mRNA expression in Ang II-free medium. EXP3174, which is an active metabolite of losartan, almost completely blunted Ang II-mediated RGS2 mRNA expression without direct stimulation of RGS2 mRNA expression. Moreover, pretreatment with olmesartan abolished Ang II-mediated RGS2 mRNA expression. Treatment with a protein kinase C inhibitor partially decreased losartan-mediated RGS2 mRNA expression. These results suggest that AT1R blockers inhibit RGS2 mRNA expression in response to Ang II via an AT1R-mediated mechanism. However, the AT1R blocker, losartan, behaves as a direct agonist for RGS2 mRNA expression via AT1R through protein kinase C-dependent and -independent pathways. In conclusion, losartan exhibits dual effects on RGS2 mRNA expression, and the direct upregulation of RGS2 mRNA expression may provide a new strategy for the treatment of hypertension.

Angiopoietin-like 7 is an anti-angiogenic protein required to prevent vascularization of the cornea.

PURPOSE: We sought to identify the anti-angiogenic molecule expressed in corneal keratocytes that is responsible for maintaining the avascularity of the cornea. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured with either human dermal fibroblasts or with human corneal keratocytes under serum-free conditions. The areas that exhibited blood vessel formation were estimated by immunostaining the cultures with an antitibody against CD31, a blood vessel marker. We also performed microarray gene-expression analysis and selected one molecule, angiopoietin-like 7 (ANGPTL7) for further functional studies conducted with the keratocytes and in vivo in mice. RESULTS: Areas showing blood vessel formation in normal serum-free medium were conditions were markedly smaller when HUVECs were co-cultured with corneal keratocytes than when they were co-cultured with the dermal fibroblasts under the same conditions. Microarray analysis revealed that ANGPTL7 expression was higher in keratocytes than in dermal fibroblasts. In vitro, inhibiting ANGPTL7 expression by using a specific siRNA led to greater tube formation than did the transfection of cells with a control siRNA, and this increase in tube formation was abolished when recombinant ANGPTL7 protein was added to the cultures. In vivo, intrastromal injections of an ANGPTL7 PshRNA into the avascular corneal stroma of mice resulted in the growth of blood vessels. CONCLUSIONS: ANGPTL7, which is abundantly expressed in keratocytes, plays a major role in maintaining corneal avascularity and transparency.