Katowice, Poland
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Wozniak A.,Medical University of Lublin | Niedziela P.,District Railway Hospital | Rejdak R.,Medical University of Lublin
Balkan Medical Journal | Year: 2014

Background: Euphrasia officinalis L. is an herb traditionally used in folk medicine, mainly in the treatment of eye disorders. Aims: The present study analyzed the activity of three extracts of E. officinalis L. (ethanol, ethyl acetate and heptane) on cultured human corneal epithelial cells (10.014 pRSV-T). Study Design: In vitro study. Methods: Toxicity, free radical scavenging activity and the immunomodulatory effects of the extracts were tested using the thiazolyl blue tetrazolium bromide (MTT) or Neutral Red, 2,2-Diphenyl- 1-picrylhydrazyl (DPPH) and ELISA tests, respectively. Moreover, nitric oxide levels and cytoskeleton architecture were analyzed after corneal cell incubation with the plant extracts. Results: We show that the biological effect depended on both the concentration and the extraction solvent used. Heptane extracts, distinct from those in ethanol and ethyl acetate, were toxic to 10.014 pRSV-T cells at low concentrations (25 μg/mL) and did not demonstrate free radical scavenging effects. All tested extracts decreased pro-inflammatory cytokine expression (IL-1β, IL-6 and TNF-α) and also anti-inflammatory IL-10 expression by human corneal cells when the extracts were added to the cell culture medium for 24 h. Conclusion: In conclusion, we show that the promising effects of the application of E. officinalis L. preparations as a supplementary therapy for eye disorders are associated with the ethanol and ethyl acetate extracts, not the heptane extract. © Trakya University Faculty of Medicine.


PubMed | Medical University of Silesia, Katowice, District Railway Hospital and University of Silesia
Type: | Journal: Computers in biology and medicine | Year: 2016

Meibomian gland dysfunction is a common cause of dry eye syndrome which can also lead to eyelid inflammation. Today, diagnostics of meibomian glands is not fully automatic yet and is based on a qualitative assessment made by an ophthalmologist. Therefore, this article proposes a new automatic analysis method which provides a quantitative assessment of meibomian gland dysfunction.The new algorithm involves a sequence of operations: image acquisition (acquisition of data from OCULUS Keratograph 5M); image pre-processing (image conversion to gray levels, median filtering, removal of uneven lighting, normalization); main image processing (binarization, morphological opening, labeling, Gaussian filtering, skeletonization, distance transform, watersheds). The algorithm was implemented in Matlab with Image Processing Toolbox (Matlab: Version 7.11.0.584, R2010b) on a PC running Windows 7 Professional, 64-bit with the Intel Core i7-4960X CPU @ 3.60GHz.The algorithm described in this article has the following features: it is fully automatic, provides fully reproducible results - sensitivity of 99.3% and specificity of 97.5% in the diagnosis of meibomian glands, and is insensitive to parameter changes. The time of image analysis for a single subject does not exceed 0.5s. Currently, the presented algorithm is tested in the Railway Hospital in Katowice, Poland.


Paduch R.,Maria Curie Sklodowska University | Jakubowicz-Gil J.,Maria Curie Sklodowska University | Niedziela P.,District Railway Hospital
Indian Journal of Experimental Biology | Year: 2010

Tumor chemoresistance and metastasis are some of the most important problems in colon cancer therapy. In the present study, co-cultures of human colon carcinoma cell spheroids, obtained from different grades of tumor, with human colon epithelium, myofibroblast and endothelial cell monolayers were performed. The purpose of these co-cultures was to reflect, in in vitro conditions, different stages of colon tumor development. In order to investigate the invasive capacities of the tumor cells and their resistance to chemotherapy, HGF, HSP27, HSP72 and MRP levels were analyzed after incubation of the co-cultures with IL-1p and irinotecan (CPT-11) added as single agents or in combination. Myofibroblasts produced significantly higher amounts of HGF than epithelial cells. Tumor cells released trace amounts of this molecule. In co-cultures, IL-1p induced HGF release, while CPT-11 alone or combined with IL-1p decreased HGF secretion. An immunoblotting analysis followed by densitometry revealed that the combination of IL-1 p plus CPT-11 added to the co-cultures led to a decrease in HSPs and MRP levels. In conclusion, direct and paracrine interactions of colon tumor cell spheroids with normal cells and exogenously added CPT-11 change HSP27, HSP72 and MRP expression in comparison to monocultures. IL-1p and CPT-11, dependent on whether they are added separately or jointly, differentially modulate HGF expression in monocultures of colon tumor spheroids or normal cells and their co-cultures.


Wojciak-Kosior M.,Medical University of Lublin | Paduch R.,Maria Curie Sklodowska University | Matysik-Wozniak A.,Medical University of Lublin | Niedziela P.,District Railway Hospital | Donica H.,Medical University of Lublin
Folia Histochemica et Cytobiologica | Year: 2011

In this article, we look at how ursolic and oleanolic acids can be used for the purpose of quality control of natural products used in dermatocosmetology as well as of various other therapeutic preparations. Ursolic acid (UA) and oleanolic acid (OA) are pentacyclic triterpenes and they are constituents of many medicinal herbs. In this study, we analyzed the cytotoxic and anti-proliferative activity of OA and UA against normal human skin fibroblasts (HSF). Additionally, the scavenging activity of free radicals of both acids was analyzed. The sensitivity of cells to OA and UA activity was determined using a standard spectrophotometric (MTT) assay. The free radical scavenging activity of OA and UA was measured using the DPPH test. The F-actin cytoskeletal proteins organization was analyzed using TRITC-phalloidine fluorescent staining. The cytotoxic activity of the analyzed acids was determined using Neutral Red (NR) uptake assay. Of the two isomeric compounds, UA showed a higher cytotoxic activity against HSF cells than did OA. Our investigations showed that OA, in view of its non-toxic nature, may be used as a supplementary factor for dermal preparations. © Polish Society for Histochemistry and Cytochemistry Folia Histochem Cytobiol. 2011.


Micali A.,Messina University | Pisani A.,Messina University | Puzzolo D.,Messina University | Nowinska A.,District Railway Hospital | And 6 more authors.
Ophthalmology | Year: 2014

Objective To demonstrate the corneal morphologic aspects obtained with in vivo confocal microscopy (CM) and light and electron microscopy of specimens obtained from the same patients with macular corneal dystrophy (MCD). Design Case series. Participants Five consecutive patients affected by MCD undergoing penetrating keratoplasty (PK) in 1 eye. Methods The patients were examined with the slit-lamp, optical pachymetry, and CM before undergoing PK. The corneal buttons were processed for light, transmission, and scanning electron microscopy. Main Outcome Measures Corneal in vivo CM, corneal light, and electron microscopy. Results Confocal microscopy showed areas of altered reflectivity in basal epithelial cells, which appeared hyperreflective or completely white. In the anterior stroma, rectilinear hyperreflective areas were shown. The stroma was characterized by a granular appearance of both keratocytes and extracellular matrix. Dark striae of different length and orientation were present in the middle and posterior stroma. The corneal endothelium showed polymegethism and cells containing bright granules in their cytoplasm. The histopathologic study demonstrated areas of thickened Bowman's layer covered by an epithelium reduced in height. The Bowman's layer thickenings were due to the accumulation of free or vesiculated material of different electron density. The keratocytes showed intracytoplasmatic vesicles, whereas the extracellular matrix presented a large quantity of intercellular electron-lucent material and parallel lamellae with an undulated course. Occasional macrophages, filled with vesicles of granular-filamentous material and evident podosomes, were observed. Descemet's membrane was formed by a normal anterior banded zone and a posterior nonbanded zone of honeycombed aspect. The endothelial cells showed a large number of intracytoplasmic vesicles. Conclusions The structural changes observed with the histopathologic methods give an account and provide an explanation for the pathologic changes demonstrated by CM in the course of MCD. This may contribute to the understanding of in vivo imaging, allowing a better, noninvasive study of the disease evolution. © 2014 by the American Academy of Ophthalmology.


Aragona P.,Messina University | Wylegala E.,District Railway Hospital | Wroblewska-Czajka E.,District Railway Hospital | Smedowski A.,District Railway Hospital | And 5 more authors.
Cornea | Year: 2014

PURPOSE:: The aim was to describe the confocal and histological findings of 2 corneas from a patient with an advanced case of type I mucopolysaccharidoses Hurler-Scheie disease (MPS IH-S). METHODS:: Both corneas from an MPS IH-S-affected patient were examined in vivo using confocal microscopy and then removed and processed for evaluation using light microscopy and transmission and scanning electron microscopy. RESULTS:: Confocal microscopy evaluation showed basal epithelial cells with either diffuse or granular hyperreflectivity. Keratocytes were highly reflective determining a web-shaped stromal appearance. Endothelial cells were barely visible. The histopathological study demonstrated superficial cells with apical microfolds, small vesicles, and evident intercellular junctions. The wing cells showed either well-evident tonofilaments and small peripheral vesicles, or large paranuclear vesicles. The basal cells showed polygonal shapes, many small vesicles, and enlarged intercellular spaces. The Bowman layer was either normal or thinner and was formed by variably electron dense material. In the stroma, irregularly oriented lamellae, many vesicle-filled keratocytes, and intercellular granular material were present. The Descemet membrane was normal, whereas the corneal endothelium showed marked degenerative changes. CONCLUSIONS:: The confocal alterations appeared consequent to the anomalous accumulation of material. The histopathological images gave a clue to better understand the corneal changes demonstrated by the confocal studies in MPS IH-S. Copyright © 2013 by Lippincott Williams & Wilkins.


To find morphometric factors predisposing to the development of corneal decompensation following cataract surgery. Study group consisted of 50 patients after keratoplasty performed as pseudophakic/aphakic corneal edema (PCE/ACE) treatment. Control group formed 50 patients after cataract removal without signs of corneal decompensation. Specific subgroups were analyzed too. The morphometric data of the fellow eye anterior chamber were obtained with Visante OCTTM. Anterior chamber depth and anterior chamber angle width were significantly smaller in study group (p < 0.00001, U-test). With excluded preoperative risk factors: Fuchs dystrophy, acute angle closure glaucoma attack history the significant differences were also observed (p < 0.01). In Fuchs' dystrophy and intraoperative complication subgroups comparison these two anterior chamber parameters were smaller too (p < 0.001). Anterior chamber width was also smaller in study group with p = 0.001. Central corneal thickness was higher in study group with p = 0.013. After exclusion of patients with Fuchs' dystrophy there was no difference in comparison (p = 0.34). The difference in total axial length comparison was insignificant (p = 0.18). Relative anterior microphthalmos was diagnosed in 31 patients of study group (62%) and in 17 of control (34%). Small anterior chamber dimensions and higher central corneal thickness due to Fuchs' dystrophy are factors influencing the risk of PCE/ACE development.


Grolik M.,Jagiellonian University | Szczubialka K.,Jagiellonian University | Wowra B.,District Railway Hospital | Dobrowolski D.,District Railway Hospital | And 3 more authors.
Journal of Materials Science: Materials in Medicine | Year: 2012

Novel polymeric hydrogel scaffolds for corneal epithelium cell culturing based on blends of chitosan with some other biopolymers such as hydroxypropylcellulose, collagen and elastin crosslinked with genipin, a natural substance, were prepared. Physicochemical and biomechanical properties of these materials were determined. The in vitro cell culture experiments with corneal epithelium cells have indicated that a membrane prepared from chitosan-collagen blend (Ch-Col) provided the regular stratified growth of the epithelium cells, good surface covering and increased number of the cell layers. Ch-Col membranes are therefore the most promising material among those studied. The performance of Ch-Col membranes is comparable with that of the amniotic membrane which is currently recommended for clinical applications. © 2012 Springer Science+Business Media, LLC.


PubMed | District Railway Hospital and Messina University
Type: Journal Article | Journal: Investigative ophthalmology & visual science | Year: 2016

To investigate corneal confocal microscopic changes in nonneoplastic and neoplastic monoclonal gammopathies.Three groups of subjects were considered: group 1, twenty normal subjects; group 2, fifteen patients with monoclonal gammopathy of undetermined significance (MGUS); group 3, eight patients with smoldering multiple myeloma and eight patients with untreated multiple myeloma. After hematologic diagnosis, patients underwent ophthalmologic exam and in vivo confocal microscopic study. The statistical analysis was performed using ANOVA and Student-Newman-Keuls tests and receiver operating characteristic (ROC) curve analysis.Epithelial cells of gammopathic patients showed significantly higher reflectivity than controls, demonstrated by optical density (P < 0.001). Subbasal nerve density, branching, and beading were significantly altered in gammopathic patients (P = 0.01, P = 0.02, P = 0.02, respectively). The number of keratocytes was significantly reduced in neoplastic patients (P < 0.001 versus both normal and MGUS) in the anterior, medium, and posterior stroma. The ROC curve analysis showed good sensitivity and specificity for this parameter. Group 2 and 3 keratocytes showed higher nuclear and cytoplasmatic reflectivity in the medium and posterior stroma. Endothelial cells were not affected.Patients with neoplastic gammopathies showed peculiar alterations of the keratocyte number, which appeared significantly reduced. A follow-up with corneal confocal microscopy of patients with MGUS is suggested as a useful tool to identify peripheral tissue alterations linked to possible neoplastic disease development.


Wylegala E.,District Railway Hospital | Nowinska A.K.,District Railway Hospital | Wroblewska-Czajka E.,District Railway Hospital | Janiszewska D.,District Railway Hospital
Indian Journal of Ophthalmology | Year: 2013

Optical coherence tomography has already been proven to be useful for pre- and post-surgical anterior eye segment assessment, especially in lamellar keratoplasty procedures. There is no evidence for intraoperative usefulness of optical coherence tomography (OCT). We present a case report of the intraoperative donor disc attachment assessment with spectral-domain optical coherence tomography in case of Descemet stripping automated endothelial keratoplasty (DSAEK) surgery combined with corneal incisions. The effectiveness of the performed corneal stab incisions was visualized directly by OCT scan analysis. OCT assisted DSAEK allows the assessment of the accuracy of the Descemet stripping and donor disc attachment.

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