While limbal epithelial cells are used for treating ocular surface wounds, the therapeutic potential of mesenchymal cells cultivated from the limbal stroma (LMSC) is less clear. We have therefore ...examined the effects of LMSC when applied to acute ocular surface wounds. LMSC derived from male rabbits (RLMSC) were applied to the ocular surface of female rabbits immediately following removal of the corneal and limbal epithelium. Human amniotic membrane (HAM) was used as the vehicle for implanting the RLMSC. The effects of RLMSC were examined when applied alone (n = 3) and in conjunction with a stratified culture of human limbal epithelial cells (HLE) grown on the opposing surface of the HAM (n = 3). Outcomes were monitored over 3 months in comparison with animals receiving no treatment (n = 3) or treatment with HLE alone on HAM (n = 3). Animals treated with RLMSC (n = 6) displayed faster re-epithelialization (∼90% versus 70% healing after 12 weeks), with best results being observed when RLMSC were pre-cultivated and implanted in the presence of HLE (p < 0.01; 90% healing by 7 weeks). While all animals displayed conjunctival cells on the corneal surface (by presence of goblet cells and/or keratin 13 expression) and corneal neovascularization, evidence of corneal epithelial regeneration was observed in animals that received RLMSC in the presence of HLE (by staining for keratin 3 and the absence of goblet cells). Conversely, corneal neovascularization was significantly greater when RLMSC were applied in the absence of HLE (<0.05; 90% of cornea compared with 20–30% in other cohorts). Nevertheless, neither human nuclear antigen nor rabbit Y chromosome were detected within the regenerated epithelium. Our results demonstrate that while cultured LMSC encourage corneal re-epithelialization, healing is improved by the pre-cultivation and implantation of these mesenchymal cells in the presence of limbal epithelial cells.
A recombinant formulation of silk fibroin containing the arginine–glycine–aspartic acid (RGD) cell-binding motif (RGD-fibroin) offers potential advantages for the cultivation of corneal cells. Thus, ...we investigated the growth of corneal stromal cells and epithelial cells on surfaces created from RGD-fibroin, in comparison to the naturally occurring Bombyx mori silk fibroin. The attachment of cells was compared in the presence or absence of serum over a 90 min period and analyzed by quantification of dsDNA content. Stratification of epithelial cells on freestanding membranes was examined by confocal fluorescence microscopy and optimized through use of low molecular weight poly(ethylene glycol) (PEG; 300 Da) as a porogen, the enzyme horseradish peroxidase (HRP) as a crosslinking agent, and stromal cells grown on the opposing membrane surface. The RGD-fibroin reduced the tendency of stromal cell cultures to form clumps and encouraged the stratification of epithelial cells. PEG used in conjunction with HRP supported the fabrication of more permeable freestanding RGD-fibroin membranes, that provide an effective scaffold for stromal–epithelial co-cultures. Our studies encourage the use of RGD-fibroin for corneal cell culture. Further studies are required to confirm if the benefits of this formulation are due to changes in the expression of integrins, components of the extracellular matrix, or other events at the transcriptional level.
The New Zealand White rabbit has been widely used as a model of limbal stem cell deficiency (LSCD). Current techniques for experimental induction of LSCD utilize caustic chemicals, or organic ...solvents applied in conjunction with a surgical limbectomy. While generally successful in depleting epithelial progenitors, the depth and severity of injury is difficult to control using chemical-based methods. Moreover, the anterior chamber can be easily perforated while surgically excising the corneal limbus. In the interest of creating a safer and more defined LSCD model, we have therefore evaluated a mechanical debridement technique based upon use of the AlgerBrush II rotating burr. An initial comparison of debridement techniques was conducted in situ using 24 eyes in freshly acquired New Zealand White rabbit cadavers. Techniques for comparison (4 eyes each) included: (1) non-wounded control, (2) surgical limbectomy followed by treatment with 100% (v/v) n-heptanol to remove the corneal epithelium (1–2 min), (3) treatment of both limbus and cornea with n-heptanol alone, (4) treatment of both limbus and cornea with 20% (v/v) ethanol (2–3 min), (5) a 2.5-mm rounded burr applied to both the limbus and cornea, and (6) a 1-mm pointed burr applied to the limbus, followed by the 2.5-mm rounded burr applied to the cornea. All corneas were excised and processed for histology immediately following debridement. A panel of four assessors subsequently scored the degree of epithelial debridement within the cornea and limbus using masked slides. The 2.5-mm burr most consistently removed the corneal and limbal epithelia. Islands of limbal epithelial cells were occasionally retained following surgical limbectomy/heptanol treatment, or use of the 1-mm burr. Limbal epithelial cells were consistently retained following treatment with either ethanol or n-heptanol alone, with ethanol being the least effective treatment overall. The 2.5-mm burr method was subsequently evaluated in the right eye of 3 live rabbits by weekly clinical assessments (photography and slit lamp examination) for up to 5 weeks, followed by histological analyses (hematoxylin & eosin stain, periodic acid-Schiff stain and immunohistochemistry for keratin 3 and 13). All 3 eyes that had been completely debrided using the 2.5-mm burr displayed symptoms of ocular surface failure as defined by retention of a prominent epithelial defect (∼40% of corneal surface at 5 weeks), corneal neovascularization (2–3 quadrants), reduced corneal transparency and conjunctivalization of the corneal surface (demonstrated by the presence of goblet cells and/or staining for keratin 13). In conclusion, our findings indicate that the AlgerBrush II rotating burr is an effective method for the establishment of ocular surface failure in New Zealand White rabbits. In particular, we recommend use of the 2.5-mm rotating burr for improved efficiency of epithelial debridement and safety compared to surgical limbectomy.
•The Algerbrush II is an effective tool for induction of LSCD in rabbits.•Optimal results are achieved using a 2.5-mm rounded burr tool.•The rabbit cornea becomes conjunctivalized and vascularized within 5 weeks.
Ocular trauma in children can result in catastrophic visual and psychological outcomes both for the child and his/her family. According to the WHO, childhood blindness is one of the major causes of ...avoidable blindness and so target of the Vision 2020 program. To achieve this program's goals, it is necessary to explore the epidemiological patterns of ocular trauma in different countries which in its turn could be a valuable means to guide us in developing preventive measures. Our study is a hospital-based prospective study which was conducted to reveal detailed information about children who were admitted to a large tertiary care hospital with the diagnosis of lacerating globe injury due to a sharp object.
During a 6-month period (December 2006 to June 2007), 125 eyes of 125 children (age ≤16 years) with the diagnosis of open-globe injury were enrolled. Clinical data were documented. Demographic data and details of the event were explored and documented based on the study questionnaire (adopting the Birmingham Eye Trauma Terminology and the United States Eye Injury Registry model as basis) by interviewing the parents and the child.
The mean age was 8.5 years (SD = 4.36 years, range: 1-16). The boys:girls ratio was 2.5:1; this ratio was age dependent, showing a significant increase with age. The home was the most frequent place (61.7%) for the trauma to occur in. Most traumas (50%) occurred in the afternoon (12 midday to 6 p.m.). Almost half of the traumas were unintentional self-injuries and occurred when the child was alone. A knife and fireworks were the two most frequent causes of injury.
As the home is the most common place of ocular trauma in this age group and as lack of proper supervision and accessibility of sharp objects such as knives are the background of the event in most cases, it is strongly recommended to educate parents about providing a safe environment at home for their children. Work-related eye injuries in this age group should also be reduced by regulations for prohibiting children's work or at least providing the necessary education and safety goggles.
The lymphovascular space invasion (LVSI) is suggested as a prognostic factor for endometrial cancer in many studies, but it has not yet been employed in FIGO staging system. The present study was ...aimed to evaluate the impact of LVSI on survival in patients with early stage endometrioid endometrial cancer.
This retrospective cohort was conducted on early stage endometrial cancer patients who underwent surgical staging total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH/BSO) and omental biopsy at Referral Teaching Hospitals of Tehran from 2005 to 2021. Patient's age, menopause status, tumor grade, tumor size, depth of myometrial invasion, LVSI and lower segment involvement were recorded. Data were analyzed with SPSS 22.
415 patients with stage I and grade 1-2, endometrioid endometrial cancer were analyzed. 100 patients (24.1%) were LVSI-positive. 3-year and 5-year survival rates were 97.1% and 88.9%, respectively. Recurrence occurred in 53 patients (12.8%). 3-year overall survival rates in LVSI-negative and LVSI-positive were 98.7% and 92%. These rates for 5-year survival were 92.1% and 79%, respectively. Recurrence rates in LVSI-negative were 8.9% while it was 25% in LVSI-positive cases. Multivariate analysis showed that LVSI has significant correlation with 3-year and 5-year overall survival rates.
LVSI in early stage endometrial cancer significantly and independently influences 3-year and 5-year survival rates and acts as a strong prognostic factor in these patients. LVSI should be implemented in endometrial cancer staging systems due to its significant correlation with cancer recurrence rates and 5-year survival rates.
Almost 13% of all death in the world is related to cancer. One of the major reasons for failing cancer treatment is the late diagnosis of the tumors. Thus, diagnosis at the early stages could be ...vital for the treatment. Serum autoantibodies, as tumor markers, are becoming interesting targets due to their medical and biological relevance. Among them, anti-p53 autoantibody in human sera is found to be involved in a variety of cancers. Regarding this issue, a novel and sensitive electrochemical biosensor for detection of anti-p53 autoantibody has been developed. For this purpose, a nanocomposite including thionine (as an electron transfer mediator)/chitosan/nickel hydroxide nanoparticles/electrochemically reduced graphene oxide (Th-CS-Ni(OH)2NPs–ERGO) as a support platform was fabricated on the surface of glassy carbon electrode via a layer-by-layer manner and characterized through common electrochemical and imaging techniques. Then, p53-antigen was immobilized on the nanocomposite and used in an indirect immunoassay with horseradish peroxidase (HRP)-conjugated secondary antibody and H2O2 as the substrate, following the typical Michaelis–Menten kinetics. Under optimized condition, two techniques, including differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS) as a label free technique, applied for the biomarker detection. The linear ranges and LODs were obtained 0.1–500 pg mL−1 and 0.001 pg mL−1 using DPV and 5–150 pg mL−1 and 0.007 pg mL−1 using EIS, respectively. Furthermore, the proposed biosensor displayed satisfying stability, selectivity, and reproducibility. According to the results, the presented protocol is promising to develop other electrochemical biosensors.
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•An electrochemical immunosensor for the determination of anti-p53 autoantibody was developed.•A nanocomposite including Th/CS/Ni(OH)2NPs/ERGO as a support platform on the surface of GC electrode has been fabricated.•Application to spiked human serum samples was successfully performed.
Determination of Human Serum Albumin (HSA), an important biomarker, is needed for early diagnosis and therapy monitoring in a variety of diseases. A novel electrochemical immunosensor was constructed ...to detect HSA in both standard solutions and human serum samples. In so doing, glassy carbon electrode (GCE) was modified in three steps. First, graphene oxide (GO) was drop-casted on the GCE surface and electrochemically reduced. Second, in situ electrochemical deposition (ECD) of cupric oxide nanoparticles (CuO NPs) in two consecutive steps has been done. Finally, anti-HSA antibodies have been immobilized via amino groups of chitosan which drop-casted earlier. The surface morphology and nature of the modified electrode was characterized using common electrochemical and imaging techniques. Using differential pulse voltammetry (DPV), the response of the immunosensor was linear in the range of 10–450 ng.mL−1 HSA, with the limit of detection (LOD) 2.6 ng.mL−1. Moreover, the selectivity of the modified electrode was investigated to some common interferences. Under optimum conditions, the immunosensor displayed excellent stability, reproducibility, selectivity and repeatability. The findings indicated that fabricated electrode is a promising candidate for quantifying HSA that is applicable in diagnostics and therapeutics.
•Reduced Graphene Oxide/CuO NPs/Chitosan Nanocomposite was successfully applied for detection of the Human Serum Albumin (HSA).•The constructed immunoassay exhibited a linear detection range from 10 to 450 ng.mL−1, with the limit of detection (LOD) 2.6 ng.mL−1.•The immunosensor performed excellently on the detection of clinical serum samples of HSA.•The constructed diagnostic device is better than other reported studies in term of LOD and dynamic linear range.
Data about the prevalence of biopsy-proven kidney diseases in Iran are rare, and none of the previous studies used electron microscopy for diagnosis. This study aimed to analyze the prevalence of ...biopsy-proven kidney diseases in Iran's primary referral center. To the best of our knowledge, this is the most extensive study carried out in Iran. Reports of kidney biopsy samples from patients referred to our center in 2007-2018 were reviewed for demographic data, clinical presentation, and final diagnosis. Statistical analyses were performed. Among the 3455 samples received, 2975 were analyzed. Nephrotic syndrome (39%) was the most common cause of biopsy, followed by subnephrotic proteinuria (18%), hematuria in association with proteinuria (15%), renal failure (9%), isolated hematuria (6%), and lupus nephritis (LN) (4%). The most common diagnoses were membranous glomerulonephritis (17.9%), focal segmental glomerulosclerosis (FSGS) (15.9%), LN (13.7%), minimal histopathological findings (unsampled FSGS vs. minimal change disease, 12.1%), Immunoglobin A nephropathy (IgAN) (6.5%) and Alport syndrome (6.1%). NS and proteinuria were the most common indications for a kidney biopsy. IgAN and LN were the most common causes of primary and secondary glomerulonephritis, presenting with hematuria and proteinuria, respectively. Although membranous glomerulonephritis was the most common disease, it has been replaced by FSGS in recent years.
Background: Developing a potent and safe scaffold is challenging in anti-cancer drug discovery. Objectives: The study focused on developing novel series of compounds based on the inhibition of ...epidermal growth factor receptor tyrosine kinase (EGFR-TK) as one of the most promising compounds in cancer therapy. Methods: In this study, a novel series of quinazoline-2,4,6-triamine derivatives were designed and synthesized through intramolecular C-H activation reaction of para-nitro aniline, trichloroacetonitrile, and isocyanides employing a one-pot reaction. Results: The in-vitro antitumor activities of the compounds which showed acceptable inhibitory effects were investigated against breast (MCF-7), lung (A-549), and colon (HT-29) cancer cell lines by employing MTT assay. All compounds had the most negligible cytotoxicity toward normal fibroblast human cell lines. Based on structural and thermodynamics analysis results, it was found that Met 769 is a key residue in interaction with all inhibitors through the formation of hydrogen bonds with high occupancies with the amine group on the quinazoline ring of inhibitors. Also, there was a good consistency between calculated ΔG binding and experimental IC50 values of compounds 10d, 10e, and erlotinib. Conclusions: Compound 10e had an extensive range of antitumor activity on three diverse cell lines comparable with erlotinib and doxorubicin reference drugs. Also, compound 10d showed selective cytotoxicity against cancerous lung cells (A-549). On the other side, computational studies confirmed that Met 769 is a crucial residue in interaction with all inhibitors.
Clear cell carcinoma (CCC) is an uncommon histopathologic subtype of ovarian and endometrial carcinoma. Due to the morphologic overlapping with other subtypes of ovarian and endometrial carcinomas, ...an accurate diagnosis is crucial.
In this study, 31 cases of ovarian clear cell carcinoma (OCCC), 28 endometrial clear cell carcinoma (ECCC), and 80 non-CCC subtypes (33 high-grade serous carcinomas of the ovary, 2 low-grade serous carcinomas, 10 ovarian endometrioid, 3 serous carcinomas and 29 endometrioid carcinomas of the endometrium) were investigated for immunohistochemical expression of AMACR. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for the distinction of OCCC and ECCC from other histopathologic subtypes were calculated.
Positive AMACR staining was seen in 18 OCCCs (58%) and 10 ECCCs (35.7%). In the non-clear cell group, 44 cases of ovarian (98%) and 25 cases of endometrial carcinoma (78%) showed negative results. Only one case of ovarian endometrioid carcinoma and 7 cases (22%) of endometrial endometrioid carcinomas revealed a positive reaction (
<0.05). Collectively, sensitivity, specificity, PPV, and NPV of AMACR expression, for the diagnosis of OCCC were calculated as 58%, 98%, 94.7%, and 77.2%, respectively. The sensitivity, specificity, PPV, and NPV were shown to be as 35.7%, 78.1%, 58.8%, and 58.1%, respectively in the endometrium.
AMACR may be a highly specific immunohistochemical marker for the distinction of serous and clear cell carcinoma. A small percentage of endometrioid carcinoma may show positive staining. The sensitivity of this marker may not be higher than the other well-known Napsin-A IHC marker.
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