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Journal of Clinical Medicine Sep 2021We have assessed the spectral transmittance of the different layers of the human cornea in the ultraviolet (UV), visible, and near-infrared (IR) spectral ranges....
We have assessed the spectral transmittance of the different layers of the human cornea in the ultraviolet (UV), visible, and near-infrared (IR) spectral ranges. Seventy-four corneal sample donors were included in the study. Firstly, the corneal transmittance was measured using a spectrophotometer. Then, all samples were fixed for histopathological analysis, which allowed us to measure the thickness of each corneal layer. Finally, the absorption coefficients of the corneal layers were computed by a linear model reproducing total transmittance. The results show that corneal transmission was almost in unity at the visible and IR ranges but not at the UV range, in which the layer with higher transmission is Descemet's membrane, whereas the stroma showed the lowest transmittance. Regarding the absorption coefficient, the most absorptive tissue was Bowman's layer, followed by the endothelium. Variations on transmittance due to changes in the stroma, Bowman's layer, or Descemet layer were simulated, and important transmission increases were found due to stroma and Bowman changes. To conclude, we have developed a method to measure the transmittance and thickness for each corneal layer. All corneal layers absorb UV light to a greater or lesser extent. The absorption coefficient is higher for Bowman's layer, while the stroma is the layer with the lowest transmittance due to its thickness. Variations in stroma thickness or changes in the corneal tissue of Bowman's layer or the endothelium layer due to some pathologies or surgeries could affect, to a greater or lesser degree, the total transmission of the cornea. Thus, obtaining accurate absorption coefficients for different layers would help us to predict and compensate these changes.
PubMed: 34640506
DOI: 10.3390/jcm10194490 -
American Journal of Physiology. Renal... Nov 2021Following our previous reports on mesangial sclerosis and vascular proliferation in diabetic nephropathy (DN) (Kriz W, Löwen J, Federico G, van den Born J, Gröne E,...
Following our previous reports on mesangial sclerosis and vascular proliferation in diabetic nephropathy (DN) (Kriz W, Löwen J, Federico G, van den Born J, Gröne E, Gröne HJ. 312: F1101-F1111, 2017; Löwen J, Gröne E, Gröne HJ, Kriz W. 317: F399-F410, 2019), we now describe the advanced stages of DN terminating in glomerular obsolescence and tubulointerstitial fibrosis based on a total of 918 biopsies. The structural aberrations emerged from two defects: ) increased synthesis of glomerular basement membrane (GBM) components by podocytes and endothelial cells leading to an accumulation of GBM material in the mesangium and ) a defect of glomerular vessels consisting of increased leakiness and an increased propensity to proliferate. Both defects may lead to glomerular degeneration. The progressing compaction of accumulated worn-out GBM material together with the retraction of podocytes out of the tuft and the collapse and hyalinosis of capillaries results in a shrunken tuft that fuses with Bowman's capsule (BC) to glomerular sclerosis. The most frequent pathway to glomerular decay starts with local tuft expansions that result in contacts of structurally intact podocytes to the parietal epithelium initiating the formation of tuft adhesions, which include the penetration of glomerular capillaries into BC. Exudation of plasma from such capillaries into the space between the parietal epithelium and its basement membrane causes the formation of insudative fluid accumulations within BC spreading around the glomerular circumference and, via the glomerulotubular junction, onto the tubule. Degeneration of the corresponding tubule develops secondarily to the glomerular damage, either due to cessation of filtration in cases of global sclerosis or due to encroachment of the insudative spaces. The degenerating tubules induce the proliferation of myofibroblasts resulting in interstitial fibrosis. Based on analysis of 918 human biopsies, essential derangement in diabetic nephropathy consists of accumulation of worn-out glomerular basement membrane in the mesangium that may advance to global sclerosis. The most frequent pathway to nephron dropout starts with the penetration of glomerular capillaries into Bowman's capsule (BC), delivering an exudate into BC that spreads around the entire glomerular circumference and via the glomerulotubular junction onto the tubule, resulting in glomerular sclerosis and chronic tubulointerstitial damage.
Topics: Biopsy; Bowman Capsule; Capillaries; Capillary Permeability; Diabetic Nephropathies; Disease Progression; Endothelial Cells; Fibrosis; Glomerular Basement Membrane; Glomerulonephritis; Humans; Microscopy, Electron, Transmission; Neovascularization, Pathologic; Nephrons; Podocytes
PubMed: 34541901
DOI: 10.1152/ajprenal.00669.2020 -
Cellular and Molecular Gastroenterology... 2022Microvillus inclusion disease (MVID) is a congenital diarrheal disorder resulting in life-threatening secretory diarrhea in newborns. Inactivating and nonsense mutations... (Review)
Review
Microvillus inclusion disease (MVID) is a congenital diarrheal disorder resulting in life-threatening secretory diarrhea in newborns. Inactivating and nonsense mutations in myosin Vb (MYO5B) have been identified in MVID patients. Work using patient tissues, cell lines, mice, and pigs has led to critical insights into the pathology of MVID and a better understanding of both apical trafficking in intestinal enterocytes and intestinal stem cell differentiation. These studies have demonstrated that loss of MYO5B or inactivating mutations lead to loss of apical sodium and water transporters, without loss of apical CFTR, accounting for the major pathology of the disease. In addition, loss of MYO5B expression induces the formation of microvillus inclusions through apical bulk endocytosis that utilizes dynamin and PACSIN2 and recruits tight junction proteins to the sites of bulk endosome formation. Importantly, formation of microvillus inclusions is not required for the induction of diarrhea. Recent investigations have demonstrated that administration of lysophosphatidic acid (LPA) can partially reestablish apical ion transporters in enterocytes of MYO5B KO mice. In addition, further studies have shown that MYO5B loss induces an imbalance in Wnt/Notch signaling pathways that can lead to alterations in enterocyte maturation and tuft cell lineage differentiation. Inhibition of Notch signaling leads to improvements in those cell differentiation deficits. These studies demonstrate that directed strategies through LPA receptor activation and Notch inhibition can bypass the inhibitory effects of MYO5B loss. Thus, effective strategies may be successful in MVID patients and other congenital diarrhea syndromes to reestablish proper apical membrane absorption of sodium and water in enterocytes and ameliorate life-threatening congenital diarrhea.
Topics: Animals; Diarrhea; Humans; Malabsorption Syndromes; Microvilli; Mucolipidoses; Myosin Type V; Sodium; Water
PubMed: 35660026
DOI: 10.1016/j.jcmgh.2022.04.015 -
PloS One 2023The steep cornea in keratoconus can greatly impair eyesight. The etiology of keratoconus remains unclear but early injury that weakens the corneal stromal architecture...
BACKGROUND
The steep cornea in keratoconus can greatly impair eyesight. The etiology of keratoconus remains unclear but early injury that weakens the corneal stromal architecture has been implicated. To explore keratoconus mechanics, we conducted two numerical simulation studies.
METHODS
A finite-element model describing the five corneal layers and the heterogeneous mechanical behaviors of the ground substance and lamellar collagen-fiber architecture in the anterior and posterior stroma was developed using the Holzapfel-Gasser-Ogden constitutive model. The geometry was from a healthy subject. Its stroma was divided into anterior, middle, and posterior layers to assess the effect of changing regional mechanical parameters on corneal displacement and maximum principal stress under intraocular pressure. Specifically, the effect of softening an inferocentral corneal button, the collagen-based tissues throughout the whole cornea, or specific stromal layers in the button was examined. The effect of simply disorganizing the orthogonally-oriented posterior stromal fibers in the button was also assessed. The healthy cornea was also subjected to eye rubbing-like loading to identify the corneal layer(s) that experienced the most tensional stress.
RESULTS
Conical deformation and corneal thinning emerged when the corneal button or the mid-posterior stroma of the button underwent gradual softening or when the collagen fibers in the mid-posterior stroma of the button were dispersed. Softening the anterior layers of the button or the whole cornea did not evoke conical deformation. Button softening greatly increased and disrupted the stress on Bowman's membrane while mid-posterior stromal softening increased stress in the anterior layers. Eye rubbing profoundly stressed the deep posterior stroma while other layers were negligibly affected.
DISCUSSION
These observations suggest that keratoconus could be initiated, at least partly, by mechanical instability/damage in the mid-posterior stroma that then imposes stress on the anterior layers. This may explain why subclinical keratoconus is marked by posterior but not anterior elevation on videokeratoscopy.
Topics: Humans; Keratoconus; Biomechanical Phenomena; Cornea; Corneal Stroma; Collagen
PubMed: 36730305
DOI: 10.1371/journal.pone.0278455 -
Immunity Nov 2020Inflammatory signaling is required for hematopoietic stem and progenitor cell (HSPC) development. Here, we studied the involvement of RIG-I-like receptors (RLRs) in HSPC...
Inflammatory signaling is required for hematopoietic stem and progenitor cell (HSPC) development. Here, we studied the involvement of RIG-I-like receptors (RLRs) in HSPC formation. Rig-I or Mda5 deficiency impaired, while Lgp2 deficiency enhanced, HSPC emergence in zebrafish embryos. Rig-I or Mda5 deficiency reduced HSPC numbers by inhibiting inflammatory signals that were in turn enhanced in Lgp2 deficient embryos. Simultaneous reduction of Lgp2 and either Rig-I or Mda5 rescued inflammatory signals and HSPC numbers. Modulating the expression of the signaling mediator Traf6 in RLR deficient embryos restored HSPC numbers. Repetitive element transcripts could be detected in hemogenic endothelial cells and HSPCs, suggesting a role as RLR ligands. Indeed, ectopic expression of repetitive elements enhanced HSPC formation in wild-type, but not in Rig-I or Mda5 deficient embryos. Manipulation of RLR expression in mouse fetal liver HSPCs indicated functional conservation among species. Thus, repetitive elements transcribed during development drive RLR-mediated inflammatory signals that regulate HSPC formation.
Topics: Animals; Biomarkers; Chromatin Assembly and Disassembly; DNA Transposable Elements; Disease Susceptibility; Gene Expression Regulation, Developmental; Hematopoiesis; Hematopoietic Stem Cells; Immunity, Innate; Immunohistochemistry; Inflammation; RNA Helicases; RNA-Binding Proteins; Repetitive Sequences, Nucleic Acid; Signal Transduction; TNF Receptor-Associated Factor 6; Valproic Acid; Zebrafish; Zebrafish Proteins
PubMed: 33159854
DOI: 10.1016/j.immuni.2020.10.007 -
Biomolecules Apr 2023Bowman's layer is an acellular layer in the anterior stroma found in the corneas of humans, most other primates, chickens, and some other species. Many other species,... (Review)
Review
Bowman's layer is an acellular layer in the anterior stroma found in the corneas of humans, most other primates, chickens, and some other species. Many other species, however, including the rabbit, dog, wolf, cat, tiger, and lion, do not have a Bowman's layer. Millions of humans who have had photorefractive keratectomy over the past thirty plus years have had Bowman's layer removed by excimer laser ablation over their central cornea without apparent sequelae. A prior study showed that Bowman's layer does not contribute significantly to mechanical stability within the cornea. Bowman's layer does not have a barrier function, as many cytokines and growth factors, as well as other molecules, such as EBM component perlecan, pass bidirectionally through Bowman's layer in normal corneal functions, and during the response to epithelial scrape injury. We hypothesized that Bowman's layer represents a visible indicator of ongoing cytokine and growth factor-mediated interactions that occur between corneal epithelial cells (and corneal endothelial cells) and stromal keratocytes that maintain the normal corneal tissue organization via negative chemotactic and apoptotic effects of modulators produced by the epithelium on stromal keratocytes. Interleukin-1 alpha, produced constitutively by corneal epithelial cells and endothelial cells, is thought to be one of these cytokines. Bowman's layer is destroyed in corneas with advanced Fuchs' dystrophy or pseudophakic bullous keratopathy when the epithelium becomes edematous and dysfunctional, and fibrovascular tissue commonly develops beneath and/or within the epithelium in these corneas. Bowman's-like layers have been noted to develop surrounding epithelial plugs within the stromal incisions years after radial keratotomy. Although there are species-related differences in corneal wound healing, and even between strains within a species, these differences are not related to the presence or absence of Bowman's layer.
Topics: Humans; Animals; Dogs; Rabbits; Epithelium, Corneal; Endothelial Cells; Corneal Stroma; Chickens; Cornea; Wound Healing; Cytokines
PubMed: 37238641
DOI: 10.3390/biom13050771 -
International Journal of Environmental... Dec 2022Membrane capacitance (C), a bioimpedance-derived measure of cell membrane health, has been suggested as an indicator of health status. However, there are few published...
Membrane capacitance (C), a bioimpedance-derived measure of cell membrane health, has been suggested as an indicator of health status. However, there are few published data to support its use in clinical settings. Hence, this study evaluated clinically relevant sources of variation, precision, and reliability of C measurements. This longitudinal study included 60 premenopausal women. Sources of variability (e.g., demographics, body composition, serum measures, diet) were identified by stepwise regression. Precision and reliability were assessed by the coefficient of variation (CV), intraclass correlation coefficients (ICC), and technical error of the measurement (TEM) for intra-day (30 min apart) and inter-day measurements (7-14 days apart). Body composition, temperature, and metabolic activity were identified as sources of variability. C measurements had high precision (CV = 0.42%) and high reliability for intra-day (ICC = 0.996) and inter-day (ICC = 0.959) measurements, independent of menstrual cycle and obesity status. Our results showed that C measurements are sensitive to clinical factors and have high precision and reliability. The results of this study suggest that C is sufficiently reliable for health status monitoring in conditions with variation in body composition, metabolic activity, or body temperature among premenopausal women.
Topics: Humans; Female; Longitudinal Studies; Reproducibility of Results; Observer Variation; Body Composition; Obesity
PubMed: 36613010
DOI: 10.3390/ijerph20010686 -
Investigative Ophthalmology & Visual... Jun 2020Quasi-static optical coherence elastography (OCE) is an emerging technology to investigate corneal biomechanical behavior in situations similar to physiological stress...
PURPOSE
Quasi-static optical coherence elastography (OCE) is an emerging technology to investigate corneal biomechanical behavior in situations similar to physiological stress conditions. Herein OCE was applied to evaluate previously inaccessible biomechanical characteristics of human corneal tissue and to study the role of Bowman's layer in corneal biomechanics.
METHODS
Human corneal donor buttons (n = 23) were obtained and Descemet's membrane and endothelium were removed. In 11 corneas, Bowman's layer was ablated by a 20 µm stromal excimer laser ablation. Buttons were mounted on an artificial anterior chamber and subjected to a pressure modulation from 10 to 30 mm Hg, and back to 10 mm Hg, in steps of 1 mm Hg. At each step, a spectral-domain optical coherence tomography scan was obtained. Displacements were analyzed by optical flow tracking, and strain over the entire stromal depth was retrieved from the phase gradient of the complex interference signal.
RESULTS
During pressure increase, corneal tissue moved upward (486-585 nm/mm Hg) but did not fully recover (Δ= 2.63 to 8.64 µm) after pressure decrease. Vertical corneal strain distribution was negative in the anterior and positive in the posterior cornea, indicating simultaneous corneal compression and expansion, respectively. Bowman's layer caused minor localized differences in corneal strain distribution.
CONCLUSIONS
Corneal strain distribution is more complex than previously assumed, with a fundamental difference in mechanical response between the anterior and posterior stroma. Clinically, OCE technology might be used to monitor the progression of corneal ectatic diseases and to determine the success of corneal cross-linking.
Topics: Biomechanical Phenomena; Cornea; Elasticity Imaging Techniques; Humans; Tomography, Optical Coherence
PubMed: 32539132
DOI: 10.1167/iovs.61.6.29 -
The American Journal of Pathology Dec 2022The development of focal and segmental glomerulosclerosis (FSGS) as a consequence of glomerular hypertension resulting from arterial hypertension is widely considered a...
The development of focal and segmental glomerulosclerosis (FSGS) as a consequence of glomerular hypertension resulting from arterial hypertension is widely considered a podocyte disease. However, the primary damage is encountered in the mesangium. In acute settings, mesangial cells disconnect from their insertions to the glomerular basement membrane, causing a ballooning of capillaries and severe changes of the folding pattern of the glomerular basement membrane, of the arrangement of the capillaries, and thereby of the architecture of the tuft. The displacement of capillaries led to contact of podocytes and parietal epithelial cells, initiating the formation of tuft adhesions to Bowman's capsule, the committed lesion to progress to FSGS. In addition, the displacement of capillaries also caused an abnormal stretching of podocytes, resulting in podocyte damage. Thus, the podocyte damage that starts the sequence to FSGS is predicted to develop secondary to the mesangial damage. This sequence was found in two hypertensive rat models of FSGS and in human hypertensive nephrosclerosis.
Topics: Rats; Humans; Animals; Podocytes; Glomerulosclerosis, Focal Segmental; Nephrosclerosis; Capillaries; Glomerular Basement Membrane; Hypertension, Renal
PubMed: 36150506
DOI: 10.1016/j.ajpath.2022.08.007 -
Investigative Ophthalmology & Visual... May 2021To identify global gene expression changes in the corneal epithelium of keratoconus (KC) patients compared to non-KC myopic controls.
PURPOSE
To identify global gene expression changes in the corneal epithelium of keratoconus (KC) patients compared to non-KC myopic controls.
METHODS
RNA-sequencing was performed on corneal epithelium samples of five progressive KC and five myopic control patients. Selected results were validated using TaqMan quantitative PCR (qPCR) on 31 additional independent samples, and protein level validation was conducted using western blot analysis on a subset. Immunohistochemistry was performed on tissue microarrays containing cores from over 100 KC and control cases. WNT10A transcript levels in corneal epithelium were correlated with tomographic indicators of KC disease severity in 15 eyes. Additionally, WNT10A was overexpressed in vitro in immortalized corneal epithelial cells.
RESULTS
WNT10A was found to be underexpressed in KC epithelium at the transcript (ratio KC/control = 0.59, P = 0.02 per RNA-sequencing study; ratio = 0.66, P = 0.03 per qPCR) and protein (ratio = 0.07, P = 0.06) levels. Immunohistochemical analysis also indicated WNT10A protein was decreased in Bowman's layer of KC patients. In contrast, WNT10A transcript level positively correlated with increased keratometry (Kmax ρ = 0.57, P = 0.02). Finally, WNT10A positively regulated COL1A1 expression in corneal epithelial cells.
CONCLUSIONS
A specific Wnt ligand, WNT10A, is reduced at the mRNA and protein level in KC epithelium and Bowman's layer. This ligand positively regulates collagen type I expression in corneal epithelial cells. The results suggest that WNT10A expression in the corneal epithelium may play a role in progressive KC.
Topics: Adult; Blotting, Western; Bowman Membrane; Collagen Type I; Collagen Type I, alpha 1 Chain; Epithelium, Corneal; Female; Gene Expression Regulation; Humans; Immunohistochemistry; Keratoconus; Male; Phenotype; Plasmids; RNA, Messenger; Real-Time Polymerase Chain Reaction; Sequence Analysis, RNA; Transcriptome; Wnt Proteins; Young Adult
PubMed: 33988693
DOI: 10.1167/iovs.62.6.16