ABSTRACT
Any opacification of the lens can be defined as cataracts, and lens epithelium cells play a crucial role in guaranteeing lens transparency by maintaining its homeostasis. Although several causative genes of congenital cataracts have been reported, the mechanisms underlying lens opacity remain unclear. In this study, a large family with congenital cataracts was collected and genetic analysis revealed a pathological mutation (c.3857 C > T, p.T1287I) in the GBF1 gene; all affected individuals in the family carried this heterozygous mutation, while unaffected family members did not. Functional studies in human lens epithelium cell line revealed that this mutation led to a reduction in GBF1 protein levels. Knockdown of endogenous GBF1 activated XBP1s in the unfolded protein response signal pathway, and enhances autophagy in an mTOR-independent manner. Heterozygous Gbf1 knockout mice also displayed typic cataract phenotype. Together, our study identified GBF1 as a novel causative gene for congenital cataracts. Additionally, we found that GBF1 deficiency activates the unfolded protein response and leads to enhanced autophagy, which may contribute to lens opacity.
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Limbal niche cells (LNCs) are one of the most important supporting cells for corneal epithelial stem cells (CES), however, research on LNCs has been mostly limited to humans and rats previously. To expand the research work into the rabbit animal model, one of the most often used animals in stem cell study, this study was carried out for the in vitro isolation and identification of rabbit LNCs. Rabbit LNCs were isolated by collagenase A digestion method and single cells were obtained, the cells were then seeded on 5% Matrigel-coated plastic surface and cultured in modified embryonic stem cell medium (MESCM). Three biological replicates of the isolating and characterization were recorded from New Zealand White rabbits aged from 2.5 months to 5 months. LNC markers (VIM/CD90/CD105/SCF/PDGFRß) were analyzed using tyramide signal amplification (TSA) staining, immunohistochemical staining (IHC), western blotting (WB), and real-time reverse transcription polymerase chain reaction (qPCR). TSA staining suggested that VIM was highly expressed in rabbit limbus stroma, which was confirmed by WB, and P63α was expressed in the basal limbus epithelium. Pan-CK and CK12 were highly expressed in the central corneal epithelium but lightly expressed in the limbal epithelium. The WB result indicated that PDGFRß and VIM expressions in rabbit-LNCs P4 were higher than in P1 and P7. In addition, rabbit corneal epithelium highly expressed Paired Box 6 (PAX6) and Epidermal growth factor-like domain 6(EGFL6). For the three repeat experiments, the cell expansion activity of rabbit-LNC was highest at P4. Rabbit-LNCs were passaged from P0 to P7, and the number of cell doublings (NCD) of P4 for the three repeat experiments was 2.816, 2.737, and 2.849. qPCR showed that high mRNA expression levels of VIM, CD90, CD105, SCF, and PDGFRß in rabbit-LNCs P4. In conclusion, rabbit-LNCs could be successfully isolated by the collagenase A digestion method as used in human tissue. There were similar characteristics between rabbit and human LNCs (VIM+/CD90+/CD105+/SCF+/PAX6+/PDGFRß+).
Subject(s)
Epithelium, Corneal , Limbus Corneae , Rabbits , Rats , Humans , Animals , Stem Cells , Cornea , Cells, Cultured , Collagenases , Epithelial Cells , Stem Cell NicheABSTRACT
Enterovirus A71 (EV-A71) is a highly contagious virus that poses a major threat to global health, representing the primary etiological agent for hand-foot and mouth disease (HFMD) and neurological complications. It has been established that interferon signaling is critical to establishing a robust antiviral state in host cells, mainly mediated through the antiviral effects of numerous interferon-stimulated genes (ISGs). The host restriction factor SHFL is a novel ISG with broad antiviral activity against various viruses through diverse underlying molecular mechanisms. Although SHFL is widely acknowledged for its broad-spectrum antiviral activity, it remains elusive whether SHFL inhibits EV-A71. In this work, we validated that EV-A71 triggers the upregulation of SHFL both in cell lines and in a mouse model. Knockdown and overexpression of SHFL in EVA71-infected cells suggested that this factor could markedly suppress EV-A71 replication. Our findings further revealed an intriguing mechanism of SHFL that it could interact with the nonstructural proteins 3Dpol of EV-A71 and promoted the degradation of 3Dpol through the ubiquitin-proteasome pathway. Furthermore, the zinc-finger domain and the 36 amino acids (164-199) of SHFL were crucial to the interaction between SHFL and EV-A71 3Dpol . Overall, these findings broadened our understanding of the pivotal roles of SHFL in the interaction between the host and EV-A71.
Subject(s)
Enterovirus A, Human , Enterovirus Infections , Enterovirus , Hand, Foot and Mouth Disease , Animals , Mice , Enterovirus A, Human/genetics , Proteasome Endopeptidase Complex , Gene Products, pol , Antigens, Viral/genetics , Antiviral Agents , Interferons , UbiquitinsABSTRACT
BACKGROUND: Mental disorders are a leading cause of disability and early mortality. The objective of this study was to describe and compare psychosocial indicators and mental health service use among ethnoculturally-diverse Ontarians. METHODS: This is a cross-sectional analysis of the Ontario Health Study pilot investigation. Residents were mailed an invitation to one of 3 assessment centres (urban, rural and northern sites) from March 2009 to July 2010. Participants had an interview with a nurse and completed a questionnaire on a touchscreen kiosk. The questionnaire included sociodemographic items, and scales assessing symptoms of depressive symptoms (CES-D) and anxiety (GAD-7), social support (Lubben Social Network Scale), stressful life events, and mental health service use. RESULTS: Eight thousand two hundred thirty-five residents participated, among whom 6652 (82.4 %) self-reported their ethnocultural background as White, 225 (2.8 %) as South Asian, 222 (2.8 %) East Asian, 214 (2.7 %) Southeast Asian, 197 (2.4 %) Black, and 28 (0.3 %) as Aboriginal. Based on their sociodemographic characteristics, participants from these ethnocultural minority groups were matched to White participants. Black participants reported significantly greater stressful life events than White participants (p = .04), particularly death (p < .05), divorce (p = .002) and financial difficulties (p < .001). East Asian participants reported significantly less social support than their White counterparts (p < .001), and this was not confounded by measurement variance. Mental health service use was significantly lower in all ethnocultural minorities except Aboriginals, when compared to White participants (p = .001). CONCLUSIONS: There is a high burden of psychosocial distress in several preponderant ethnocultural minorities in Ontario; many of whom are not accessing available mental health services.
Subject(s)
Ethnicity/statistics & numerical data , Health Services Accessibility/statistics & numerical data , Mental Health/ethnology , Minority Groups/statistics & numerical data , Adult , Black or African American/psychology , Asian People/psychology , Black People/psychology , Cross-Sectional Studies , Female , Health Status , Humans , Male , Mental Health Services/statistics & numerical data , Middle Aged , Ontario , Rural Population/statistics & numerical data , Social Support , White People/psychologyABSTRACT
In anterior cruciate ligament (ACL) revision cases, the resultant bigger aperture at the tibia footprint can cause graft instability. The increased movement hinders bone-graft integration and leads to graft abrasion. This article describes a technique to optimize graft stability when using a soft tissue graft for ACL revision. The technique is used when there is suspicion of size mismatch between the new tibia footprint aperture and the graft. The first stage involves passing a suture via an anterolateral tibial tunnel connecting with the revision tibia tunnel distal to the tibia footprint aperture. The new graft is subsequently deployed, and the potential discrepancy between graft diameter and aperture is confirmed. The second stage involves placing 2 pulling sutures on the new graft and passing them into the anterolateral tibial tunnel. The tensioned and anchored pulling sutures secure graft stability at the tibia footprint, and the graft distal to that is fixed routinely. The lasso technique stabilizes the new graft at the tibia footprint by tensioning it in a distal and anterolateral direction. For selected cases, this technique enables a 1-stage ACL revision with a soft tissue graft when faced with graft instability at the tibia footprint.
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Purpose: To test the effects and underlying mechanisms of basic fibroblast growth factor (bFGF) on the limbal niche cell (LNC) function ex vivo. Methods: By using different concentrations of bFGF (0, 4, 8, 12, and 16 ng/mL) and fibroblast growth factor receptor (FGFR) inhibitors, the effects of bFGF on LNC proliferation, expression of stem cell markers, and transcription levels of the ß-catenin were investigated. Single-cell RNA sequencing (scRNA-seq) was used to analyze the action and mechanisms of FGFR subtypes and the Wnt/ß-catenin pathway during LNC culture. An mature corneal epithelial cell (MCEC)/LNC three-dimensional model was constructed to verify whether bFGF activates the Wnt/ß-catenin pathway in LNC by inhibiting FGFR or ß-catenin targets. Results: scRNA-seq showed that FGFR1 is the main receptor in LNC, along with the molecules in the Wnt pathway, including WNT2, FZD7, LRP5, LRP6, and ß-catenin. The 12 ng/mL bFGF treatment group showed higher LNC proliferation rate and transcription levels of OCT4, SOX2, NANOG, and ß-catenin than any other groups (P < 0.001). In the MCEC/LNC co-culture model, MCEC/LNC treated with 12 ng/mL bFGF promoted the aggregation of the spheres than other groups, associated with increased transcription levels of P63α, WNT2, ß-catenin, and a decreased transcription level of CK12 (P < 0.001). Wnt/ß-catenin inhibitor LF3 treatment reversed the abovementioned effect of bFGF. Conclusions: bFGF could maintain and promote the stemness of LNC via the FGFR1/Wnt2/FZD7/LRP6 axis in a concentration-dependent manner.
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BACKGROUND: Lipocalin 13 (LCN13) is a member of the lipocalin family that consists of numerous secretory proteins. LCN13 high-expression has been reported to possess anti-obesity and anti-diabetic effects. Although metabolic dysfunction-associated steatotic liver diseases (MASLD) including metabolic dysfunction-associated steatohepatitis (MASH) are frequently associated with obesity and insulin resistance, the functional role of endogenous LCN13 and the therapeutic effect of LCN13 in MASH and related metabolic deterioration have not been evaluated. METHODS: We employed a methionine-choline deficient diet model and MASH cell models to investigate the role of LCN13 in MASH development. We sought to explore the effects of LCN13 on lipid metabolism and inflammation in hepatocytes under PA/OA exposure using Western blotting, real-time RT-PCR, enzyme-linked immunosorbent assay, hematoxylin and eosin staining, oil red O staining. Using RNA sequencing, chromatin immunoprecipitation assay, and luciferase reporter assays to elucidate whether farnesoid X receptor (FXR) regulates human LCN13 transcription as a transcription factor. RESULTS: Our study found that LCN13 was down-regulated in MASH patients, MASH mouse and cell models. LCN13 overexpression in hepatocyte cells significantly inhibited lipid accumulation and inflammation in vitro. Conversely, LCN13 downregulation significantly exacerbated lipid accumulation and inflammatory responses in vivo and in vitro. Mechanistically, we provided the first evidence that LCN13 was transcriptionally activated by FXR, representing a novel direct target gene of FXR. And the key promoter region of LCN13 binds to FXR was also elucidated. We further revealed that LCN13 overexpression via FXR activation ameliorates hepatocellular lipid accumulation and inflammation in vivo and in vitro. Furthermore, LCN13-down-regulated mice exhibited aggravated MASH phenotypes, including increased hepatic lipid accumulation and inflammation. CONCLUSION: Our findings provide new insight regarding the protective role of LCN13 in MASH development and suggest an innovative therapeutic strategy for treating MASH or related metabolic disorders.
Subject(s)
Carcinoma, Hepatocellular , Fatty Liver , Liver Neoplasms , Animals , Humans , Mice , Carcinoma, Hepatocellular/metabolism , Fatty Liver/metabolism , Inflammation/metabolism , Lipids , Lipocalins/metabolism , Liver , Liver Neoplasms/metabolism , Mice, Inbred C57BL , Obesity/metabolismABSTRACT
Introduction Open reduction internal fixation (ORIF) and primary arthrodesis are two conventional options for the treatment of Lisfranc injuries. However, they are associated with implant-related complications. An alternative suspensory device construct using interosseous nonabsorbable sutures with endobuttons has been described with satisfactory results. This study aims to explore functional outcomes after suture button fixation of Lisfranc injuries in a Southeast Asian population. Methods This was a single-surgeon retrospective study of patients with Lisfranc injuries treated surgically using a suture button fixation technique between 2017 and 2019. Data collected included demographic information, pre-injury levels of activity, nature of injury, and type of surgery performed. The minimum postoperative follow-up was one year. The Foot and Ankle Outcome Score (FAOS) and Foot and Ankle Ability Measure (FAAM) were used to evaluate patient-reported outcomes. Scores were reported in percentage (%) with median and interquartile range. Results Twenty-nine patients with a mean age of 29 years (21-76) were recruited. Sixteen underwent suture button fixation only (SB), and 13 underwent suture button fixation with intercuneiform screw fixation and plating (SBM). The median scores for the FAOS and FAAM questionnaires were at least 80% in all domains. Twenty-eight patients (97%) were able to return to pre-injury activity level, 27 patients (93%) were able to return to sports. Only one patient was not satisfied with the outcomes of surgery. No patients had post-traumatic arthritis or hardware failure necessitating implant removal at the final follow-up. Conclusion This study has demonstrated that treatment of Lisfranc injuries with a suspensory device construct resulted in good outcomes with 97% of patients being able to return to pre-injury activity levels, and 93% of patients being able to return to sports. It may not be necessary to perform primary arthrodesis in uncomplicated Lisfranc injuries. This technique is also advantageous as implant removal is not routinely required due to the design and biomechanical properties of suspensory devices.
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Here we report a standard procedure for the isolation and identification of limbal niche cells (LNCs). Limbus tissue obtained from an eye bank was used for LNCs isolation. The tissue was divided into 12 pieces under aseptic conditions and digested for 18 h at 37 °C in the cell culture incubator using collagenase A to obtain cell clusters with LNCs and limbal epithelial progenitor cells. The cell clusters were further digested for 15 min at 37 °C using 0.25% trypsin-EDTA to obtain single cells and then cultured in modified embryonic stem cell medium (MESCM) on a plastic surface coated with 5% Matrigel. Cells were passaged upon 70% confluence, and LNCs were identified using immunofluorescence, real-time quantitative PCR (qPCR), and flow cytometry. Primary LNCs were isolated and passaged more than 12 times. The proliferation activity of LNCs from P4 to P6 was the highest. LNCs expressed higher stem cell markers than BMMSCs (SCF, Nestin, Rex1, SSEA4, CD73, CD90, MSX1, P75NTR, and PDGFRß). Furthermore, results showed that P4 LNCs uniformly expressed VIM, CD90, CD105, and PDGFRß, but not Pan-CK, which could be used as a marker for the identification of LNCs. Flow cytometric analysis showed that approximately 95%, 97%, 92%, and 11% of LNCs expressed CD73, CD90, CD105, and SCF respectively, while they were 68%, 99%, 20%, and 3% in BMMSCs. The standard process for LNC isolation and identification could provide a reliable laboratory basis for the widespread use of LNCs.
Subject(s)
Epithelium, Corneal , Limbus Corneae , Stem Cells , Cell Culture Techniques , Cell Separation/methods , Fluorescent Antibody Technique , Cells, Cultured , Cell Differentiation , Epithelial Cells , Stem Cell NicheABSTRACT
OBJECTIVES: Fungal keratitis (FK) is a kind of serious corneal infection and penetrating keratoplasty (PKP) is needed when medical therapy fails. Although Nectria haematococca is found as endophytes in the roots of some plant species, there has been no report of N. haematococca infection in human. METHODS: We reviewed 46 patients who underwent PKP due to FK in our hospital from July 2021 to December 2021, and there were three patients who had relapsed. The next-generation sequencing revealed that all three corneas were infected with N. haematococca. RESULTS: Based on the ocular manifestation and treatment course of three cases, we summarize the characteristics of N. haematococca FK: the scope of corneal infection was widespread with severe hypopyon. The effect of local use of fluconazole and voriconazole was not ideal, and PKP was the main treatment. Even after a large-scale corneal lesion resection, the lesion may recur. The recurrence occurred primarily in the second week after PKP. CONCLUSION: This is the first clinical report of N. haematococca infection in humans. Compared with the other currently known FK caused by the Fusarium solani species complex, N. haematococca keratitis is more severe and more likely to recur.
Subject(s)
Eye Infections, Fungal , Fusarium , Keratitis , Humans , Keratitis/diagnosis , Keratitis/drug therapy , Keratitis/microbiology , Eye Infections, Fungal/diagnosis , Eye Infections, Fungal/drug therapy , Eye Infections, Fungal/microbiology , High-Throughput Nucleotide Sequencing , Antifungal Agents/therapeutic use , Retrospective StudiesABSTRACT
OBJECTIVES: This study aimed to determine the epidemiological and genetic features of human metapneumovirus (HMPV) infection in children in southern China, and the effect of meteorological factors on infection. METHODS: 14,817 children (≤14 years) with acute respiratory tract infections from 2010 to 2019 were examined for HMPV and other respiratory viruses by real-time quantitative polymerase chain reaction. Full-length F gene of 54 positive samples were sequenced and subjected to phylogenetic analysis. The correlation between the HMPV-positive rate and meteorological factors was analyzed by linear regression analysis. RESULTS: HMPV was detected in 524 (3.5%) children, who were mostly younger than 1 year. The seasonal peak of HMPV prevalence mainly occurred in spring. Respiratory syncytial virus was the most common virus coinfected with HMPV (5.3%). Phylogenetic analysis revealed that the sequenced HMPV strains belonged to four sublineages, including A2b (1.9%), A2c (31.5%), B1 (50.0%), and B2 (16.7%). After adjusting for all meteorological factors, sunshine duration was inversely correlated with the HMPV-positive rate. CONCLUSION: HMPV is an important respiratory pathogen that causes acute respiratory tract infections in children in southern China, particularly in children ≤5 years old. The prevalence peak of HMPV in this area appeared in spring, and the predominant subtype was B1. Meteorological factors, especially long sunshine duration, might decrease the HMPV prevalence.
Subject(s)
Metapneumovirus , Paramyxoviridae Infections , Respiratory Tract Infections , Child , Humans , Infant , Child, Preschool , Metapneumovirus/genetics , Retrospective Studies , Molecular Epidemiology , Phylogeny , Paramyxoviridae Infections/epidemiology , Respiratory Tract Infections/epidemiology , China/epidemiology , Meteorological ConceptsABSTRACT
BACKGROUND: In 2006, the Canadian Cardiovascular Society (CCS) Access to Care Working Group recommended a 30-day wait time benchmark for cardiac rehabilitation (CR). The objectives of the current study were to: (1) describe cardiac patient perceptions of actual and ideal CR wait times, (2) describe and compare cardiac specialist and CR program perceptions of wait times, as well as whether the recommendations are appropriate and feasible, and (3) investigate actual wait times and factors that CR programs perceive to affect these wait times. METHODS: Postal and online surveys to assess perceptions of CR wait times were administered to CR enrollees at intake into 1 of 8 programs, all CCS member cardiac specialists treating patients indicated for CR, and all CR programs listed in Canadian directories. Actual wait times were ascertained from the Canadian Cardiac Rehabilitation Registry. The design was cross-sectional. Responses were described and compared. RESULTS: Responses were received from 163 CR enrollees, 71 cardiac specialists (9.3% response rate), and 92 CR programs (61.7% response rate). Patients reported that their wait time from hospital discharge to CR initiation was 65.6 ± 88.4 days (median, 42 days), while their ideal median wait time was 28 days. Most patients (91.5%) considered their wait to be acceptable, but ideal wait times varied significantly by the type of cardiac indication for CR. There were significant differences between specialist and program perceptions of the appropriate number of days to wait by most indications, with CR programs perceiving shorter waits as appropriate (p < 0.05). CR programs reported that feasible wait times were significantly longer than what was appropriate for all indications (p < 0.05). They perceived that patient travel and staff capacity were the main factors negatively affecting waits. The median wait time from referral to program initiation was 64 days (mean, 80.0 ± 62.8 days), with no difference in wait by indication. CONCLUSIONS: Wait times following access to cardiac rehabilitation are prolonged compared with consensus recommendations, and yet are generally acceptable to most patients. Wait times following percutaneous coronary intervention in particular may need to be shortened. Future research is required to provide an evidence base for wait time benchmarks.
Subject(s)
Cardiac Rehabilitation , Cardiology , Medical Staff, Hospital/psychology , Patient Satisfaction , Specialization , Waiting Lists , Aged , Cross-Sectional Studies , Female , Health Care Surveys , Health Services Accessibility , Humans , Male , Middle Aged , Registries , Time FactorsABSTRACT
Purpose: (1) To measure the corneal endothelium-Descemet membrane (EDM) layer thickness in Descemet membrane detachment (DMD) patients in vivo using high-definition optical coherence tomography (HD-OCT), and to investigate its correlation with age. (2) To explore whether the detachment time will affect the EDM thickness. (3) To explore whether the EDM thickness of cornea with DMD was different from that without DMD. Participants: Patients with DMD were divided into three groups. Group 1 included twenty-three patients whose Descemet membrane (DM) was partial or complete detached from the corneal stroma after various ocular surgeries. Group 2 included eight patients from group 1 who underwent twice HD-OCT examination on different days before the DM reattached to the stroma. Group 3 included nine patients from group 1 who had clear grayscale boundary between the DM and stroma in HD-OCT images after DM reattachment. Methods: All patients underwent HD-OCT and EDM thickness was measured using Image -Pro Plus 6.0. In Group 1, regression analyses were used to evaluate the correlation between EDM thickness and age, and the thickness difference between the ≤50-year-old group and the >50-year-old group was analyzed by independent sample t-test. In Group 2, paired samples t-test was used to check whether detachment time would affect EDM thickness. In Group 3, paired samples t-test was used to check whether the EDM thickness of cornea with DMD was different from that without DMD. p < 0.05 was considered significant. Results: In Group 1, the EDM thickness measured on the first post-operative day was 27.8 ± 3.6 µm, and a positive correlation was found between EDM thickness and age (r = 0.619, p < 0.05). The EDM thickness of ≤50-year-old group and >50-year-old group were 23.9 ± 3.2 and 29.2 ± 2.6 µm, and there was a significant difference between the two groups (p = 0.001). In Group 2, the first measurement of EDM thickness was 27.5 ± 4.0 µm, the second measurement was 27.6 ± 4.2 µm, the interval between the two measurements was 2.1 ± 1.6 days, and there was no significant difference between the two measurements (p = 0.328). In Group 3, the EDM thickness with DM detachment was 28.3 ± 3.5 µm, with DM reattachment was 23.4 ± 2.4 µm, there was a significant difference between the two measurements (p = 0.002). Conclusions: The EDM thickness in the state of DMD is thicker than its actual thickness in normal cornea, and EDM thickness of the >50-year-old group is much thicker than that of the ≤50-year-old group.
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Constrained by the existing scaffold inability to mimic limbal niche, limbal bio-engineered tissue constructed in vitro is challenging to be widely used in clinical practice. Here, a 3D nanofiber-aerogel scaffold is fabricated by employing thermal cross-linking electrospinned film polycaprolactone (PCL) and gelatin (GEL) as the precursor. Benefiting from the cross-linked (160 °C, vacuum) structure, the homogenized and lyophilized 3D nanofiber-aerogel scaffold with preferable mechanical strength is capable of refraining the volume collapse in humid vitro. Intriguingly, compared with traditional electrospinning scaffolds, the authors' 3D nanofiber-aerogel scaffolds possess enhanced water absorption (1100-1300%), controllable aperture (50-100 µm), and excellent biocompatibility (optical density value, 0.953 ± 0.021). The well-matched aperture and nanostructure of the scaffolds with cells enable the construction of limbal bio-engineered tissue. It is foreseen that the proposed general method can be extended to various aerogels, providing new opportunities for the development of novel limbal bio-engineered tissue.
Subject(s)
Nanofibers , Gelatin , Nanofibers/chemistry , Polyesters/chemistry , Tissue Engineering/methods , Tissue Scaffolds/chemistryABSTRACT
Face masks, which serve as personal protection equipment, have become ubiquitous for combating the ongoing COVID-19. However, conventional electrostatic-based mask filters are disposable and short-term effective with high breathing resistance, causing respiratory ailments and massive consumption (129 billion monthly), intensifying global environmental pollution. In an effort to address these challenges, the introduction of a piezoelectric polymer was adopted to realize the charge-laden melt-blown via the melt-blowing method. The charge-laden melt-blown could be applied to manufacture face masks and to generate charges triggered by mechanical and acoustic energy originated from daily speaking. Through an efficient and scalable industrial melt-blown process, our charge-laden mask is capable of overcoming the inevitable electrostatic attenuation, even in a high-humidity atmosphere by long-wearing (prolonging from 4 to 72 h) and three-cycle common decontamination methods. Combined with outstanding protective properties (PM2.5 filtration efficiency >99.9%), breathability (differential pressure <17 Pa/cm2), and mechanical strength, the resultant charge-laden mask could enable the decreased replacement of masks, thereby lowering to 94.4% of output masks worldwide (â¼122 billion monthly) without substituting the existing structure or assembling process.
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Purpose: To investigate the phenotypic changes of mature corneal epithelial cells (MCECs) that cocultured with limbal niche cells (LNCs) in three-dimensional Matrigel (3D Matrigel) in vitro. Methods: MCECs were isolated from central corneas, and limbal epithelial progenitor cells (LEPCs) were isolated from limbal segments with Dispase II. LNCs were isolated and cultured from limbal niche using the collagenase A digestion method and identified with PCK/VIM/CD90/CD105/SCF/PDGFRß. MCECs were cultured on 3D Matrigel (50%, v/v) with or without LNCs for 10 days. Expression of CK12 and p63α and clone formation test were used to compare the progenitor phenotypic changes for MCECs before and after induction using LEPCs as control. Results: Homogeneous LNCs were isolated and identified as spindle shape and adherent to a plastic surface coated with 5% Matrigel. Double immunostaining of the fourth-passage LNCs was uniformly PCK-/VIM+/CD90+/CD105+/SCF+/PDGFRß+. Reverse transcription and quantitative real-time polymerase chain reaction (RT-qPCR) revealed the decrease of PCK expression from the second passage and elevation of Vim, CD90, CD105, SCF, and PDGFRß transcripts from the third passage, and the transcription level of Vim, CD90, CD105, SCF, and PDGFRß was elevated statistically in the fourth passage compared to the first passage (P < 0.01). Both immunofluorescence (IF) staining for cross section and cytospin cells demonstrated that MCECs expressed higher CK12 while lower p63α than LEPCs (P < 0.01). Sphere growth formation was noticed as early as 24 hours in the MCEC + LNC group, 48 hours in the LEPC group, and 72 hours in the MCEC group. The diameters of the spheres were the biggest in the MCEC + LNC group (182.24 ± 57.91 µm), smaller in the LEPC group (125.71 ± 41.20 µm), and smallest in the MCEC group (109.39 ± 34.85 µm) by the end of the 10-day culture (P < 0.01). Double immunostaining with CK12/p63α showed that cells in the sphere formed from MCECs expressed CK12 but not p63α; in contrast, some cells in the MCEC + LNC group expressed CK12, but most of them expressed p63α. RT-qPCR revealed a significant reduction of CK12 transcript but elevation of p63α, Oct4, Nanog, Sox2, and SSEA4 (P < 0.05). Holoclone composed of cubic epithelial cells could be generated in the MCEC + LNC group but not in the other two groups. Conclusions: The data shows that human MCEC cell phenotype could be induced to the dedifferentiation stage when cocultured with LNCs in 3D Matrigel that simulated the microenvironment of limbal stem cells in vitro.
Subject(s)
Limbus Corneae , Cell Differentiation , Collagen , Drug Combinations , Epithelial Cells/metabolism , Laminin/metabolism , ProteoglycansABSTRACT
Photosynthetic bacteria have flexible metabolisms and strong environmental adaptability, and require cheap, but plentiful, energy supplements, which all enable their use in Cr(VI)-remediation. In this study, the effects of culture conditions on the total Cr removal rate were investigated for a newly identified strain of Rhodobacter sphaeroides SC01. The subcellular distribution and Cr(VI) reduction ability of four different cellular fractions were evaluated by scanning electron microscopy and transmission electron microscopy. Experiments indicated that the optimal culture conditions for total Cr removal included a culture temperature of 35 °C, pH of 7.20, an NaCl concentration of 5 g L-1, a light intensity of 4000 lx, and an initial cell concentration (OD680) of 0.15. In addition, most Cr was found in the cell membrane in the form of Cr (III) after reduction, while cell membranes had the highest Cr(VI) reduction rate (99%) compared to other cellular components. In addition, the physical and chemical properties of SC01 cells were characterized by FTIR, XPS, and XRD analyses, confirming that Cr was successfully absorbed on bacterial cell surfaces. CrPO4â§6H2O and Cr5(P3O10)3 precipitates were particularly identified by XRD analysis. After screening supplementation with five phosphor salts, Cr(VI) reduction due to bioprecipitation was improved by the addition of Na4P2O7 and (NaPO3)6 salts, with the Cr(VI)-reduction rate combined with Na4P2O7 addition being 15% higher than that of the control. Thus, this study proposes a new Cr(VI)-removal strategy based on the combined use of photosynthetic bacteria and phosphor salts, which importantly increases its potential application in treating wastewater.
Subject(s)
Chromium , Water Pollutants, Chemical , Bacteria , Chromium/analysis , Dietary Supplements , Hydrogen-Ion Concentration , Salts , WastewaterABSTRACT
STUDY DESIGN: A systematic review and meta-analysis. OBJECTIVES: Pulmonary dysfunction is often advocated among the indications for surgical correction of adolescent idiopathic scoliosis (AIS). Previous studies have discussed the effect of scoliosis correction on respiratory function without reaching a definitive conclusion: Some showed that the respiratory function can improve after scoliosis surgery without defining the precise role of anterior, posterior, and combined approaches on this improvement; furthermore, the majority of these studies did not take normal growth into account. As a result, the role of surgery remains to be clarified. The object of the present study was to synthesize the current knowledge regarding changes in respiratory function after posterior corrective surgery for AIS. METHODS: A comprehensive systematic search was performed to identify all relevant studies in the following electronic databases: MEDLINE, EMBASE, CINAHL (EBSCO). We focused on the studies (1) that discussed posterior fusion surgery for AIS without thoracoplasty, (2) that discussed comparisons of pre- and postoperative percent-predicted values of forced vital capacity (%FVC) or forced expiratory volume (%FEV), and (3) with minimum 2-year follow-up. Forest plots were depicted and Z value was calculated as a test for overall effect. RESULTS: Ten studies (6 prospective and 4 retrospective studies) met our inclusion criteria. The overall effect showed that there was no significant difference in %FVC or %FEV between pre- and postoperative measurements (very low evidence). CONCLUSIONS: Posterior correction surgery for mild to moderate AIS patients showed no significant improvement of postoperative respiratory function measured by relative, percent-predicted values at minimum 2-year follow-up.
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PURPOSE: The Canadian Cardiovascular Society initiated a pan-Canadian process for development of quality indicators (QIs) for cardiac rehabilitation (CR). Before implementation, the QIs underwent pilot testing to ensure they were acceptable and feasible for field implementation. The objectives of this test were to assess (1) the technical feasibility of measuring the QIs; (2) the workload required to measure the QIs; and (3) acceptability of measuring the QIs and issues with their implementation. METHODS: The 2 indicators chosen for field testing were QI-1 (% of eligible inpatients referred) and 2b (median wait time from CR referral receipt to enrollment). The approach consisted of 3 steps: (1) data extraction to test technical feasibility; (2) completing a workload diary; and (3) providing input through a semistructured interview regarding acceptability and implementation issues. Three academic CR sites were selected to undertake the field test. RESULTS: QI-1 ranged from 51.0% to 68.4%, and QI-2b was reported as 27 days (median) by one site, and 22 days (mean) by another. It was not considered feasible for CR programs to assess all potentially CR-eligible inpatients for CR referral exclusions. Compilation required 4.2 hours for QI-1 and 1.8 hours for QI-2b. QI assessment was acceptable to the programs, but changes in practice would be needed at each site to implement the QIs. CONCLUSIONS: CR programs may require enhancement of information-tracking processes to enable QI measurement. It was recommended that the QIs be implemented, but should undergo minor revisions to enhance feasibility.
Subject(s)
Cardiac Rehabilitation , Eligibility Determination/statistics & numerical data , Quality Assurance, Health Care , Quality Indicators, Health Care/organization & administration , Time-to-Treatment/statistics & numerical data , Canada/epidemiology , Cardiac Rehabilitation/methods , Cardiac Rehabilitation/standards , Cardiovascular Diseases/epidemiology , Feasibility Studies , Humans , Quality Assurance, Health Care/methods , Quality Assurance, Health Care/standards , Waiting ListsABSTRACT
OBJECTIVES: We sought to describe temporal trends in the sociodemographic and clinical characteristics of participants referred to cardiac rehabilitation (CR), and its effect on programme participation and all-cause mortality over 14â years. SETTING: A large CR centre in Toronto, Canada. PARTICIPANTS: Consecutive patients between 1996 and 2010. PRIMARY AND SECONDARY OUTCOME MEASURES: Referrals received were deterministically linked to administrative data, to complement referral form abstraction. Out-of-hospital deaths were identified using vital statistics. Patients were tracked until 2012, and mortality was ascertained. Percentage attendance at prescribed sessions was also assessed. RESULTS: There were 29,171 referrals received, of which 28,767 (98.6%) were successfully linked, of whom 22,795 (79.2%) attended an intake assessment. The age of the referred population steadily increased, with more females, less affluent and more single patients referred over time (p<0.001). More patients were referred following percutaneous coronary intervention and less following coronary artery bypass graft surgery (p<0.001). The number of comorbidities decreased (p<0.001). Hypertension increased over time (p<0.001), yet the control of cholesterol steadily improved over time. The proportion of smokers decreased over time (p<0.001). Participation in CR significantly declined, and there were no significant changes in mortality. 3-year mortality rates were less than 5%. CONCLUSIONS: Characteristics of referred patients tended to reflect broader trends in risk factors and cardiovascular disease burden. Physicians appear to be referring more sociodemographically diverse patients to CR; however, programmes may need to better adapt to engage these patients to fully participate. More complex patients should be referred, using explicit criteria-based referral processes.