A newly identified gene Ahed plays essential roles in murine haematopoiesis.

The development of haematopoiesis involves the coordinated action of numerous genes, some of which are implicated in haematological malignancies. However, the biological function of many genes remains elusive and unknown functional genes are likely to remain to be uncovered. Here, we report a previously uncharacterised gene in haematopoiesis, identified by screening mutant embryonic stem cells. The gene, 'attenuated haematopoietic development (Ahed)', encodes a nuclear protein. Conditional knockout (cKO) of Ahed results in anaemia from embryonic day 14.5 onward, leading to prenatal demise. Transplantation experiments demonstrate the incapacity of Ahed-deficient haematopoietic cells to reconstitute haematopoiesis in vivo. Employing a tamoxifen-inducible cKO model, we further reveal that Ahed deletion impairs the intrinsic capacity of haematopoietic cells in adult mice. Ahed deletion affects various pathways, and published databases present cancer patients with somatic mutations in Ahed. Collectively, our findings underscore the fundamental roles of Ahed in lifelong haematopoiesis, implicating its association with malignancies.

Virtual Reality Images of the Home Are Useful for Patients With Hospital-Based Palliative Care: Prospective Observational Study With Analysis by Text Mining.

Background: Malignancy patients who need long-term hospitalization can feel loneliness affecting their quality of life. The global COVID-19 pandemic has caused visiting restrictions that could mean patients who might be missing out on family support and palliative care, therefore, need to adapt and change. We used virtual reality (VR) technology with the aim of reducing feelings of loneliness among these patients. Objectives: In a small cohort setting, we aimed to clarify the usefulness of VR viewing for this purpose by text mining interviews with the patients in palliative care after their VR experience, and to clarify the feasibility of this program. Design and Setting/Subjects: Four consecutive Japanese patients in the palliative care unit viewed personalized familiar persons or places through VR goggles, while communicating by telephone. After the VR experience, text mining of the patients' interviews was used to extract the words for the frequency count and co-occurrence analysis. Results: Four clusters were extracted: "relief from the pain of hospitalization by feeling safe and secure with family members nearby," "using VR to regain daily life," "immersive feeling of being in the same space as family," and "loneliness due to the realistic feeling of separation from the family through VR experience." There were no cases of VR sickness. Conclusion: Our results attained by text mining suggest the promising potential of VR imaging of familiar surroundings for patients in palliative care.

Origin, prospective identification, and function of circulating endothelial colony-forming cells in mice and humans.

Most circulating endothelial cells are apoptotic, but rare circulating endothelial colony-forming cells (C-ECFCs), also known as blood outgrowth endothelial cells, with proliferative and vasculogenic activity can be cultured; however, the origin and naive function of these C-ECFCs remains obscure. Herein, detailed lineage tracing revealed murine C-ECFCs emerged in the early postnatal period, displayed high vasculogenic potential with enriched frequency of clonal proliferative cells compared with tissue-resident ECFCs, and were not committed to or derived from the BM hematopoietic system but from tissue-resident ECFCs. In humans, C-ECFCs were present in the CD34bright cord blood mononuclear subset, possessed proliferative potential and in vivo vasculogenic function in a naive or cultured state, and displayed a single cell transcriptome sharing some umbilical venous endothelial cell features, such as a higher protein C receptor and extracellular matrix gene expression. This study provides an advance for the field by identifying the origin, naive function, and antigens to prospectively isolate C-ECFCs for translational studies.

Impact of rigid gas-permeable contact lens on keratometric indices and corneal thickness of keratoconus eyes examined with anterior segment optical coherence tomography.

PURPOSE/AIM: Detecting keratoconus (KC) progression helps determine the surgical indication for corneal cross-linking (CXL). This retrospective observational study aimed to examine changes in keratometric indices and corneal thickness in patients with KC who used rigid gas-permeable (RGP) contact lenses. MATERIALS AND METHODS: This study involved 31 eyes (31 patients) diagnosed with KC. No patient had used RGP or any other type of contact lenses for at least 1 month. Corneal topographic data were obtained using three-dimensional anterior segment optical coherence tomography before and after >1 month of RGP lens use. RESULTS: The average and maximum keratometry values changed after using an RGP lens (-1.05 ± 1.92 D, p < 0.01 and -1.65 ± 4.20 D, p = 0.04, respectively); the spherical component of the anterior corneal surface became significantly smaller (p = 0.02). No change was observed in the central or thinnest corneal thickness values. Keratometric changes were greater in eyes with severe KC than in those with moderate KC (p = 0.014). CONCLUSIONS: Keratometry and spherical components of the anterior corneal surface values decreased after RGP lens use; keratometric changes were greater in eyes with severe KC than in those with moderate KC. Corneal progression indices, including corneal thickness, posterior keratometry, and irregular astigmatism values, mostly remained unchanged. It is important to consider these findings when evaluating corneal topography of KC and preparing CXL.

Three-year results of accelerated transepithelial cross-linking (30 mW/cm2 × 3 min) for keratoconus: a prospective study.

OBJECTIVE: To assess the long-term efficacy and safety of accelerated transepithelial corneal cross-linking (ATE-CXL) with 30 mW/cm2 × 3 min. METHODS AND ANALYSIS: Thirty-four eyes of 23 patients with progressive keratoconus (KCN) recruited within a single centre were enrolled in this prospective interventional study. Exclusion criteria included: history of Descemet's membrane rupture, glaucoma, uveitis, severe dry eye, concurrent corneal infections, and systemic disease that could affect corneal healing. ATE-CXL was performed with 3 min of ultraviolet-A continuous irradiation (30 mW/cm2). Follow-up examinations were scheduled on postoperative day 1; 1 and 2 weeks; 1, 3 and 6 months; and 1, 2 and 3 years. Main outcome measures were maximum corneal power (Kmax), average corneal power (AvgK), steepest corneal power (Ks), central corneal thickness, thinnest corneal thickness, uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BCVA) and endothelial cell density. RESULTS: Mean Kmax, AvgK, Ks, UCVA, BCVA and endothelial cell density did not significantly change over 3 years. The speed of progression obtained by linear regression analysis on corneal parameters (Kmax, AvgK, Ks) improved after ATE-CXL. All baseline parameters correlated with the postoperative Kmax slope. Two eyes underwent ATE-CXL redo because of continued progression after the primary CXL. CONCLUSION: This is the first report of 3-year results of ATE-CXL with 30 mW/cm2 × 3 min. ATE-CXL (30 mW/cm2 × 3 min) was safe and effective for slowing down KCN progression. TRIAL REGISTRATION NUMBER: This study was registered with ID UMIN000009372 in UMIN-Clinical Trials Registry.

Comparison of efficacy and safety of accelerated trans-epithelial crosslinking for keratoconus patients with corneas thicker and thinner than 380µm.

PURPOSE: Accelerated trans-epithelial cross-linking (ATE-CXL), a therapy to halt keratoconus progression, has the merit of widening the indications for thinner corneas (<380 µm). Since a hypotonic solution affects the swollen cornea, corneas of <380 µm thickness at preoperative measurement can be an indication for ATE-CXL. The aim of this retrospective study was to compare the efficacy and safety of ATE-CXL for keratoconus between corneas with thicknesses <380 µm and ≥380 µm. MATERIALS AND METHODS: Thirty-four eyes of 27 patients who underwent ATE-CXL (30 mW/cm2; 3 minutes) with completion of a 24-month follow-up, were enrolled and divided into two groups: Group 1, thinnest corneal thickness (TCT), <380 µm (n = 10) and Group 2, TCT, ≥380 µm (n = 24). A hypotonic solution was administered to Group 1 until the corneal thickness increased by >380 µm before UV-A irradiation. We measured uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), maximum and average keratometric values (Kmax and AveK), central corneal thickness (CCT), TCT by anterior segment optical coherence tomography, and corneal endothelial cell density (ECD) using specular microscopy. The changes from baseline to 24 months postoperatively between the two groups were compared accordingly. RESULTS: The changes in Kmax and AveK from baseline to 24 months in Group 1 (ΔKmax: -7.8 ± 7.7 D, ΔAveK: -4.3 ± 6.1 D) showed significant decreases compared to those in Group 2 (ΔKmax: 0.2 ± 3.0 D, ΔAveK: 0.6 ± 2.7 D) (p = .004 and p = .001), and there were no significant changes from baseline to 24 months postoperatively in UCVA, BCVA, CCT, TCT, and ECD in both groups. CONCLUSION: ATE-CXL is effective and safe for keratoconic corneas in both groups. The effect of reducing keratometric values was greater in the group with thinner corneas.

Differences in mineral accumulation and gene expression profiles between two metal hyperaccumulators, Noccaea japonica and Noccaea caerulescens ecotype Ganges, under excess nickel condition.

Here we compare mineral accumulation and global gene expression patterns between two metal hyperaccumulator plants - Noccaea japonica, originating from Ni-rich serpentine soils, and Noccaea caerulescens (ecotype Ganges), originating from Zn/Pb-mine soils - under excess Ni conditions. Significant differences in the accumulation of K, P, Mg, B, and Mo were explained by the expression levels of specific transporters for each mineral. We previously showed that total Ni accumulation in the whole plant is higher in N. caerulescens than in N. japonica. Here we found a similar tendency for Fe under excess Ni; however, the expression of iron-regulated transporter 1 (IRT1), which encodes the primary Fe uptake transporter and causes excess Ni uptake in Arabidopsis thaliana, was higher in N. japonica. NjIRT1 has a point mutation at Asp100, which is essential for Fe transport, and so might lack its Fe and possibly Ni transport function. Noccaea japonica might have lost its IRT1 function, which would prevent excess Ni uptake via IRT1 in Ni-rich soils, and come to rely on other transporters.

Descemet stripping endothelial keratoplasty after cytomegalovirus corneal endotheliitis and immunosuppression for Mooren's ulcer.

PURPOSE: The aim of this report was to describe a case of cataract surgery and Descemet stripping automated endothelial keratoplasty (DSAEK) after cytomegalovirus (CMV) corneal endotheliitis and bullous keratopathy (BK) following immunosuppressive treatment for Mooren's ulcer. OBSERVATIONS: A 64-year-old man was referred to our hospital because of peripheral ulcerative keratitis in his left eye. He had a history of trabeculectomy for open angle glaucoma in his left eye. He was diagnosed with Mooren's ulcer and treated with topical betamethasone and tacrolimus with systemic cyclosporine. The corneal ulcer improved, but the peripheral cornea thinned from 6 to 12 and 0-2 o'clock. Five months later, cells were observed in the left anterior chamber, and real-time polymerase chain reaction examination of the aqueous humor showed CMV-DNA-positive results. The patient was diagnosed with CMV corneal endotheliitis, and oral ganciclovir was administered. Fifteen months after the initial presentation, BK appeared with decreased vision to 20 cm/n. d. After confirmation of negative CMV-DNA in the aqueous humor, DSAEK was performed following cataract surgery. The postoperative visual acuity recovered to 0.3. Mooren's ulcer exacerbation and CMV corneal endotheliitis did not recur postoperatively. CONCLUSIONS AND IMPORTANCE: This is the first report of a case in which a patient with Mooren's ulcer developed BK due to CMV corneal endotheliitis and required DSAEK. Cataract surgery and DSAEK could be performed without issue by creating the main wound and side ports in a manner that avoids the thinned parts of the cornea.

Iridotrabecular and Iridocorneal Contact Changes after Cataract Surgery and Endothelial Keratoplasty in Bilateral Iridoschisis.

We report a case of bilateral iridoschisis with corneal oedema and a quantitative evaluation of the changes in iridotrabecular and iridocorneal contact before and after cataract surgery and after Descemet stripping automated endothelial keratoplasty (DSAEK). A 76-year-old woman with iridoschisis and cataracts, previously managed with laser iridotomy, experienced progressive vision loss. The preoperative iridotrabecular contact (ITC) index measured by anterior segment optical coherence tomography was 23.6% in the right eye and 24.4% in the left eye. Preoperative corneal oedema in the right eye was more severe than that in the left eye. Cataract surgery, followed by DSAEK, was performed in the right eye and subsequently in the left eye. Her visual acuity improved postoperatively, and the corneal oedema of both eyes was treated successfully. Moreover, the ITC index improved in both eyes, to 4.7 and 6.9% after cataract surgery and to 0 and 0% after DSAEK in the right and left eyes, respectively. Staged cataract surgery and DSAEK were effective for endothelial decompensation caused by iridoschisis. Additionally, we confirm that iridotrabecular and iridocorneal contacts improved after both surgical procedures not only after cataract surgery but also after DSAEK. This case report showed the clinical usefulness of the ITC index in the detection of changes after different surgical procedures.

Assessment of Long-Term Anterior and Posterior Topographic Changes in the Cornea After Ptosis Surgery Using Fourier Harmonic Analysis.

PURPOSE: We aimed to assess the corneal refractive changes induced by ptosis surgery in patients with acquired ptosis using Fourier harmonic analysis. METHODS: This retrospective observational study enrolled consecutive patients who underwent levator aponeurotic surgery for acquired ptosis at the Department of Ophthalmology in the University of Tokyo Hospital from May 2016 to January 2018. Best corrected visual acuity, central corneal thickness, average keratometric corneal power (AvgK), corneal astigmatism, and topographic data using Fourier analysis were analyzed preoperatively and 6 months postoperatively. RESULTS: Thirty-two eyes of 32 patients (age, 72.6 ± 8.5 years) were included in this study. There were no significant differences in best corrected visual acuity and central corneal thickness. However, there were significant decreases in anterior AvgK, anterior corneal astigmatism, and posterior corneal astigmatism 6 months postoperatively (all, P < 0.001). Fourier harmonic analysis showed that the anterior spherical component significantly decreased 6 months postoperatively (P < 0.001). There were no significant differences in other components of the anterior and posterior cornea. There was a significant negative correlation between preoperative posterior AvgK and changes in posterior AvgK (r = -0.891, P < 0.001) and between preoperative posterior corneal astigmatism and changes in posterior corneal astigmatism at 6 months (r = -0.858, P < 0.001). CONCLUSIONS: Anterior and posterior corneal keratometry and posterior corneal astigmatism significantly changed 6 months after ptosis surgery for acquired ptosis.

Corneal topography in keratoconus evaluated more than 30 years after penetrating keratoplasty: a Fourier harmonic analysis.

The aim of this observational study was to examine the characteristics of anterior and posterior corneal topography in keratoconic eyes more than 30 years after penetrating keratoplasty (PK). Patients who maintained clear grafts for more than 30 years after PK were included and divided into the keratoconus (KC) group or other diseases (Others) group, based on the primary indication. Twenty-six eyes of 26 patients were included. The KC group and the Others group included 14 eyes and 12 eyes, respectively. The KC group participants were younger at the time of surgery (P = 0.03). No differences were found in best-spectacle-corrected visual acuity, keratometric power, and central-corneal-thickness. Based on corneal topography using Fourier harmonic analyses, regular astigmatism in the anterior cornea was significantly larger (P = 0.047) and the spherical component in the posterior cornea was significantly lower (P = 0.01) in the KC group. The area under the receiver operating characteristic curve of the spherical component, regular astigmatism, asymmetry component, and higher-order irregularity were 66.07%, 63.10%, 57.14%, and 59.23%, respectively, in the anterior cornea and 80.65%, 52.98%, 63.10%, and 63.99%, respectively, in the posterior cornea. Our results suggested that Fourier harmonic analysis of corneal topography could be useful for patients with KC long after PK.

Elevated Expression of Vacuolar Nickel Transporter Gene IREG2 Is Associated With Reduced Root-to-Shoot Nickel Translocation in Noccaea japonica.

A number of metal hyperaccumulator plants, including nickel (Ni) hyperaccumulators, have been identified in the genus Noccaea. The ability to accumulate Ni in shoots varies widely among species and ecotypes in this genus; however, little is known about the molecular mechanisms underlying this intra- and inter-specific variation. Here, in hydroponic culture, we compared Ni accumulation patterns between Noccaea japonica, which originated in Ni-enriched serpentine soils in Mt. Yubari (Hokkaido, Japan), and Noccaea caerulescens ecotype Ganges, which originated in zinc/lead-mine soils in Southern France. Both Noccaea species showed extremely high Ni tolerance compared with that of the non-accumulator Arabidopsis thaliana. But, following treatment with 200 µM Ni, N. caerulescens showed leaf chlorosis, whereas N. japonica did not show any stress symptoms. Shoot Ni concentration was higher in N. caerulescens than in N. japonica; this difference was due to higher efficiency of root-to-shoot Ni translocation in N. caerulescens than N. japonica. It is known that the vacuole Ni transporter IREG2 suppresses Ni translocation from roots to shoots by sequestering Ni in the root vacuoles. The expression level of the IREG2 gene in the roots of N. japonica was 10-fold that in the roots of N. caerulescens. Moreover, the copy number of IREG2 per genome was higher in N. japonica than in N. caerulescens, suggesting that IREG2 expression is elevated by gene multiplication in N. japonica. The heterologous expression of IREG2 of N. japonica and N. caerulescens in yeast and A. thaliana confirmed that both IREG2 genes encode functional vacuole Ni transporters. Taking these results together, we hypothesize that the elevation of IREG2 expression by gene multiplication causes the lower root-to-shoot Ni translocation in N. japonica.

Comparison of 5-Year Clinical Results of Descemet and Non-Descemet Stripping Automated Endothelial Keratoplasty.

PURPOSE: To compare 5-year clinical results of non-Descemet stripping automated endothelial keratoplasty (nDSAEK) and DSAEK. METHODS: A retrospective chart review involving patients with non-Fuchs-type bullous keratoplasty who underwent nDSAEK or DSAEK 5 years before the review was performed on demographics, graft survival rate, endothelial cell density (ECD), best-corrected visual acuity (BCVA), and complications. RESULTS: Fifty eyes (nDSAEK 28, DSAEK 22) were included in this study. Common indications for surgery were bullous keratopathy due to glaucoma, cataract surgeries, or laser iridotomy. The graft survival rates in nDSAEK and DSAEK eyes at 1, 3, and 5 years were 0.75 and 0.86, 0.67 and 0.81, and 0.59 and 0.74, respectively. The mean donor ECDs of nDSAEK and DSAEK eyes were 2638 and 2503 cells/mm, respectively; these decreased to 1654 and 1406, 1503 and 1218, and 1108 and 1020 cells/mm at 1, 3, and 5 years, respectively. The mean preoperative BCVAs of nDSAEK and DSAEK eyes were 1.08 and 1.11 LogMAR, respectively; these improved to 0.238 and 0.190, 0.126 and 0.157, and 0.097 and 0.070 LogMAR at 1, 3, and 5 years, respectively. The most common complication was intraocular pressure elevation. There were no statistically significant differences between nDSAEK and DSAEK in graft survival rates, changes in ECD and BCVA, or the occurrence of postoperative complications, with the exception of the change in ECD at 2 years (1173 and 1193 cells/mm, P = 0.0159). CONCLUSIONS: Our 5-year clinical results of nDSAEK and DSAEK did not show significant differences at almost all points.

Descemet Stripping Automated Endothelial Keratoplasty for Failed Penetrating Keratoplasty: Influence of the Graft-Host Junction on the Graft Survival Rate.

PURPOSE: To investigate the clinical results of Descemet stripping automated endothelial keratoplasty (DSAEK) for failed penetrating keratoplasty (PK) and the influence of the graft-host junction (GHJ) on the graft survival rate. METHODS: Data were retrospectively collected on patient demographics, visual outcomes, complications, and graft survival rate for 17 eyes of 16 patients who underwent DSAEK for failed PK. The graft survival rate was compared between the eyes when divided into a bump group and a well-aligned group according to the shape of the GHJ detected on anterior segment optical coherence tomography. RESULTS: The most common indication for initial PK was bullous keratopathy after glaucoma surgery (35.3%). Seven eyes (41.2%) were classified into the bump group and 10 eyes (58.8%) into the well-aligned group. The mean best-ever documented visual acuity (BDVA) after DSAEK was 0.33 logMAR. Postoperatively, almost 70% of eyes achieved a BDVA that was within 0.2 logMAR of their preoperative BDVA. Graft detachment occurred in 29.4% of eyes and primary graft failure in 17.6%. All primary failures occurred in the bump group. The cumulative graft survival rate was 82.3% at 1 year, 73.2% at 2 years, and 58.6% at 3 years. Graft failure was more likely in eyes in the bump group than in those in the well-aligned group (P = 0.037, Wilcoxon test). CONCLUSIONS: DSAEK for failed PK had a favorable outcome in this study. However, the GHJ should be assessed carefully before performing the procedure.

Chromatin states shape insertion profiles of the piggyBac, Tol2 and Sleeping Beauty transposons and murine leukemia virus.

DNA transposons and retroviruses are versatile tools in functional genomics and gene therapy. To facilitate their application, we conducted a genome-wide insertion site profiling of the piggyBac (PB), Tol2 and Sleeping Beauty (SB) transposons and the murine leukemia virus (MLV) in mouse embryonic stem cells (ESCs). PB and MLV preferred highly expressed genes, whereas Tol2 and SB preferred weakly expressed genes. However, correlations with DNase I hypersensitive sites were different for all vectors, indicating that chromatin accessibility is not the sole determinant. Therefore, we analysed various chromatin states. PB and MLV highly correlated with Cohesin, Mediator and ESC-specific transcription factors. Notably, CTCF sites were correlated with PB but not with MLV, suggesting MLV prefers smaller promoter-enhancer loops, whereas PB insertion encompasses larger chromatin loops termed topologically associating domains. Tol2 also correlated with Cohesin and CTCF. However, correlations with ESC-specific transcription factors were weaker, suggesting that Tol2 prefers transcriptionally weak chromatin loops. Consistently, Tol2 insertions were associated with bivalent histone modifications characteristic of silent and inducible loci. SB showed minimum preference to all chromatin states, suggesting the least adverse effect on adjacent genes. These results will be useful for vector selection for various applications.

Longitudinal Study of Age-Related Cataract Using Dynamic Light Scattering: Loss of α-Crystallin Leads to Nuclear Cataract Development.

PURPOSE: To conduct a longitudinal study on age-related nuclear cataracts using dynamic light scattering (DLS) to determine if cataract progression is associated with loss of the unbound form of the lens molecular chaperone protein, α-crystallin. DESIGN: Natural history and cohort study. PARTICIPANTS: Patients 30 years of age or older of either gender seeking treatment at the Wilmer Eye Institute Cornea-Cataract Department. METHODS: All patients underwent a comprehensive dilated eye examination every 6 months, including slit-lamp grading of their lenses using the Age-Related Eye Disease Study (AREDS) clinical lens grading system and obtaining an estimate of unbound α-crystallin level in the nucleus, the α-crystallin index (ACI), using the National Aeronautics and Space Administration-National Eye Institute DLS device. We used a random effects statistical model to examine the relationship of lens opacity changes over time with ACI changes. MAIN OUTCOME MEASURES: α-Crystallin Index (ACI) and AREDS nuclear cataract grade. RESULTS: Forty-five patients (66 eyes) 34 to 79 years of age with AREDS nuclear lens grades of 0 to 3.0 were followed up every 6 months for a mean of 19 months (range, 6-36 months). We found that lenses with the lowest baseline levels of ACI had the most rapid progression of cataracts, whereas lenses with higher ACI at baseline had no or slower cataract progression. Lenses that lost α-crystallin at the highest rates during the study also had faster progression of nuclear cataracts than lenses with a slower rate of ACI loss. Kaplan-Meier survival curves showed that lenses with the lowest initial ACI had the highest risk of undergoing cataract surgery. CONCLUSIONS: This longitudinal study corroborates our previous cross-sectional study finding that higher levels of unbound α-crystallin as assessed by ACI are associated with lower risk of cataract formation and that loss of ACI over time is associated with cataract formation and progression. This study suggested that assessment of ACI with the DLS device could be used as a surrogate for lens opacity risk in clinical studies, and for assessing nuclear cataract events in studies where cataract development may be a side effect of a drug or device.

Induction of Nickel Accumulation in Response to Zinc Deficiency in Arabidopsis thaliana.

Excessive accumulation of nickel (Ni) can be toxic to plants. In Arabidopsis thaliana, the Fe²âº transporter, iron (Fe)-regulated transporter1 (IRT1), mediates Fe uptake and also implicates in Ni²âº uptake at roots; however, the underlying mechanism of Ni²âº uptake and accumulation remains unelucidated. In the present study, we found that zinc (Zn) deficient conditions resulted in increased accumulation of Ni in plants, particularly in roots, in A. thaliana. In order to elucidate the underlying mechanisms of Ni uptake correlating zinc condition, we traced 63Ni isotope in response to Zn and found that (i) Zn deficiency induces short-term Ni²âº absorption and (ii) Zn²âº inhibits Ni²âº uptake, suggesting competitive uptake between Ni and Zn. Furthermore, the Zrt/Irt-like protein 3 (ZIP3)-defective mutant with an elevated Zn-deficient response exhibited higher Ni accumulation than the wild type, further supporting that the response to Zn deficiency induces Ni accumulation. Previously, expression profile study demonstrated that IRT1 expression is not inducible by Zn deficiency. In the present study, we found increased Ni accumulation in IRT1-null mutant under Zn deficiency in agar culture. These suggest that Zn deficiency induces Ni accumulation in an IRT1-independen manner. The present study revealed that Ni accumulation is inducible in response to Zn deficiency, which may be attributable to a Zn uptake transporter induced by Zn deficiency.

Development and evaluation of porcine atelocollagen vitrigel membrane with a spherical curve and transplantable artificial corneal endothelial grafts.

PURPOSE: To develop a collagen vitrigel (CV) optimized as a corneal endothelial cell (CEC) carrier and create an artificial corneal endothelial graft. METHODS: We first developed a flat-shaped collagen vitrigel for regenerative medicine (CV-RM) using porcine atelocollagen and ultraviolet (UV) irradiation. The optimal UV amount was determined by measuring the CV-RM transparency under various irradiating conditions. The collagen vitrigel for corneal endothelial regenerative treatment (CV-CERT), a transparent porcine atelocollagen with a curved shape, was made using spherically curved molds and UV irradiation. The membrane permeability of the CV-CERT was tested in vitro. The biocompatibility, transparency, and adhesiveness of the CV-CERT were evaluated in rabbit eyes. We also developed a culture technique for distributing human CECs on the curved CV-CERT. RESULTS: The optimal amount of UV irradiation for CV-RM transparency was 2400 mJ/cm(2). Membrane permeability of CV-CERT at day 5 was higher than that of commercially available CV (P = 0.032). The CV-CERT was transparent and biocompatible in rabbit corneas for up to 4 months. The CV-CERT remained attached to the rabbit corneal posterior surface, whereas the flat-shaped CV-RM, differing only in shape from the CV-CERT, dislocated soon after surgery. Human CECs seeded on the CV-CERT using our technique were evenly distributed with a single layer structure and a mean cell density of 2650 ± 100 cells/mm(2). CONCLUSIONS: We developed a transparent and biocompatible porcine-derived atelocollagen vitrigel membrane with a spherical curvature. A transplantable artificial endothelial graft was created by combining cultured human CECs and the CV-CERT.

Stromal bed quality and endothelial damage after femtosecond laser cuts into the deep corneal stroma.

AIM: To evaluate the stromal bed quality and endothelial damage after femtosecond laser (FSL) cuts into the deep corneal stroma. METHODS: Using a 150-kHz FSL, a lamellar cut was aimed at a depth of 100, 300, or 500 µm in porcine corneas. Stromal bed smoothness was graded from light microscopy and scanning electron microscopy images. Rabbit corneas were cut at remaining thicknesses of 70, 100 and 150 µm using the FSL. The effects of peeling off the corneal flap and the distance between laser spots (2 or 4 µm) were examined. RESULTS: The ratio of damaged cells in the group with a remaining depth of 70 µm was significantly larger than that in the groups with a remaining depth of 150 µm. The ratio of damaged cells in the group with a 4-µm spot separation and the flap peeled off was significantly larger than that in the group with a 4-µm spot separation and the flap not peeled off. CONCLUSIONS: Corneal endothelial damage is likely to increase when the remaining depth is less than 70 µm, and peeling off the flap damages corneal endothelial cells when the remaining depth is less than 100 µm.

The impact of systemic vascular resistance on the accuracy of the FloTrac/Vigileo™ system in the perioperative period of cardiac surgery: a prospective observational comparison study.

FloTrac/Vigileo™ system is based on arterial pressure waveform analysis arterial pressure-based CO (APCO). Therefore, systemic vascular resistance (SVR) can influence the accuracy of APCO. The purpose of this study is to evaluate the relationship between SVR and the accuracy of APCO. We managed 50 consecutive patients in the perioperative period of cardiac surgery with FloTrac/Vigileo™ system (v. 3.02) and Swan-Ganz catheter/Vigilance™ system pulmonary artery catheter-based CO (PAC-CO) simultaneously. Continuous hemodynamic measurement using both methods was performed every 20 s from the induction of anesthesia to PAC removal 4 h after extubation. A total of 11,092 (intraoperative), 38,455 (postoperative, pre-extubation), and 44,235 (postoperative, post-extubation) data pairs were finally analyzed. Bland-Altman analysis revealed that in the intraoperative [postoperative pre-extubation, post-extubation] period, the bias was 0.5 [0.1, 0.0] L/min and the limits of agreement ranged from -2.4 to 3.3 [-2.2 to 2.4, -2.4 to 2.3] L/min. The percentage error was 60.3 [54.5, 48.5] %. Regression analysis of the systemic vascular resistance index (SVRI) and the bias between APCO and PAC-CO showed that the bias was positively correlated to the SVRI. Subanalysis based on SVR with Lin's concordance correlation coefficient revealed that relatively satisfactory concordance was found in the normal-SVR group (concordance correlation coefficient ρ c = 0.51-0.56) regardless of vasoactive agent use. The accuracy of the FloTrac/Vigileo™ System (v. 3.02) is relatively satisfactory in the condition with normal SVR regardless of vasoactive agent use. Positive correlation between the bias and SVR can be the clue to the more effective use of FloTrac/Vigileo™ system.