Autologous serum eye drops (ASEDs) are used as a treatment for severe dry eye disease. The concentration and stability of various growth factors in ASEDs is determinative for their efficiency. We therefore assessed the concentrations of transforming growth factor beta 1 (TGF-β1), epidermal growth factor (EGF) and insulin-like growth factor 1 (IGF-1) in ASEDs following storage at 4-8, -20, -80 and -156 °C. Twenty % and 100% sera from eight healthy volunteers were analysed by the sandwich enzyme immunoassay at different time intervals up to seven months. The mean levels of TGF-β1 and EGF in undiluted and 20% serum did not differ significantly from the baseline levels in fresh serum for any storage conditions after 7 days at 4-8 °C, as well as after 4- and 7-month preservation at sub-zero temperatures. In 20% serum, no IGF-1 concentration decrease was found following 7 days of preservation at 4-8 °C. However, a decrease to 78 % and 81 % (P < 0.01) of baseline values was found in 20% serum after 4-month storage at -20 °C and 7-month storage at -156 °C, respectively. A more pronounced decrease in IGF-1 was observed in undiluted serum. All assessed growth factors present in 20% frozen serum remained stable for up to 7 months. The highest stability was achieved at -80 °C. At -20 and -156 °C, some decrease in IGF-1 occurred. Our results indicate that 20% ASEDs can be stored frozen up to 7 months under proper conditions.
PURPOSE: To assess whether the micronucleus cytome assay (MCyt) reliably detects DNA damage occurring in control and pathological superficial epithelial cells from human conjunctiva. METHODS: Impression cytology samples from the bulbar conjunctiva of 33 healthy controls, eight patients with conjunctival intraepithelial neoplasia (CIN) and eight with mucous membrane pemphigoid (MMP) were examined using the MCyt modified for the ocular surface. RESULTS: The mean number of micronuclei (MNi) in control samples was 0.94 MNi/1000 epithelial cells, with no significant difference between conjunctival quadrants and independent of sex and age. The MCyt assay applied to CIN-affected eyes showed a significantly higher frequency of MNi (18.63/1000 cells), apoptotic cells, nuclear enlargement, multinucleated cells, and keratolysis compared with the corresponding unaffected paired eyes and with the control value. Although the mean MNi frequency in MMP eyes was also higher (1.73 MNi/1000 cells), it did not prove to be statistically different from the control samples. On the other hand, the MMP-affected eyes revealed significantly elevated percentages of cells with snake-like chromatin, multinucleated cells, apoptotic cells, and nuclear buds compared with controls. CONCLUSIONS: Micronucleus cytome assay was adapted as a rapid screening test for genomic instability on the ocular surface. We have determined reference levels for MNi and other nuclear alterations on healthy conjunctiva and demonstrated that particularly frequencies of MNi are significantly elevated in conjunctiva affected by CIN. We demonstrate that MNi are more specific than other nuclear abnormalities and thus can be used for screening of ocular surface neoplasia.
- MeSH
- buněčné jádro MeSH
- epitelové buňky * MeSH
- konjunktiva MeSH
- lidé MeSH
- mikrojaderné testy MeSH
- poškození DNA * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The aim of this study was to determine the effect of autologous serum (AS) eye drops on the density of human leucocyte antigen (HLA)-DR-positive epithelial cells and Langerhans cells on the ocular surface of patients with bilateral severe dry eye disease (DED) due to graft-versus-host disease (GvHD) or Sjögren's syndrome (SS). The study was conducted on 24 patients (48 eyes). AS was applied 6-10 times daily for 3 months together with regular artificial tear therapy. HLA-DR-positive cells were detected by direct immunocytochemistry on upper bulbar conjunctiva imprints obtained before and after treatment. The application of AS drops led to a statistically significant increase in the mean density of aberrant HLA-DR-positive conjunctival epithelial cells (p < 0.05) and HLA-DR-positive Langerhans cells (p < 0.05) in the GvHD group. Aberrant HLA-DR-positive epithelial cells in the SS group were decreased non-significantly. All patients reported a significant decrease in the Ocular Surface Disease Index (p < 0.01), which indicates improvement of the patient's subjective feelings after therapy. There was an expected but non-significant decrease of aberrant HLA-DR-positive conjunctival epithelial cells in the SS group only. However, the increased density of HLA-DR-positive cells, indicating slight subclinical inflammation, does not outweigh the positive effect of AS in patients with DED from GvHD.
- MeSH
- dospělí MeSH
- epitel metabolismus MeSH
- epitelové buňky účinky léků metabolismus MeSH
- HLA-DR antigeny metabolismus MeSH
- imunohistochemie metody MeSH
- konjunktiva účinky léků metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- nemoc štěpu proti hostiteli farmakoterapie metabolismus MeSH
- oční roztoky terapeutické užití MeSH
- počet buněk metody MeSH
- senioři MeSH
- sérum metabolismus MeSH
- Sjögrenův syndrom farmakoterapie metabolismus MeSH
- syndromy suchého oka farmakoterapie metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- kmenové buňky metabolismus patologie MeSH
- lidé MeSH
- limbus corneae metabolismus patologie MeSH
- nemoci rohovky genetika metabolismus patologie MeSH
- rohovkový epitel metabolismus patologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- dopisy MeSH
- práce podpořená grantem MeSH
BACKROUND: In vivo corneal confocal microscopy allows the examination of each layer of the cornea in detail and the identification of pathological changes at the cellular level. The purpose of this study was to identify the possible effects of a three-month treatment with autologous serum eye-drops in different corneal layers of patients with severe dry eye disease using corneal confocal microscopy. METHODS: Twenty-six patients with dry eye disease were included in the study. Corneal fluorescein staining was performed. The corneas of the right eyes were examined using in vivo corneal confocal microscopy before and after a three-month treatment with autologous serum drops. The densities of superficial and basal epithelial cells, Langerhans cells, the keratocytes and activated keratocytes, the density of endothelial cells and the status of the sub-basal nerve plexus fibres were evaluated. RESULTS: A significant decrease in corneal fluorescein staining was found after the three-month autologous serum treatment (p = 0.0006). The basal epithelial cell density decreased significantly (p = 0.001), while the density of superficial epithelial cells did not change significantly (p = 0.473) nor did the number of Langerhans cells or activated keratocytes (p = 0.223; p = 0.307, respectively). There were no differences in the other corneal cell layers or in the status of the nerve fibres. CONCLUSIONS: The results demonstrate the ability of corneal confocal microscopy to evaluate an improvement in the basal epithelial cell layer of the cornea after autologous serum treatment in patients with dry eye disease. More studies with longer follow-up periods are needed to elucidate the suitability of corneal confocal microscopy to follow the effect of autologous serum treatment on nerve fibres or other corneal layers in dry eye disease patients.
- MeSH
- aplikace lokální MeSH
- fluorescein farmakologie MeSH
- fluorescenční barviva farmakologie MeSH
- konfokální mikroskopie metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- oční roztoky aplikace a dávkování MeSH
- počet buněk MeSH
- prospektivní studie MeSH
- rohovka patologie MeSH
- sérum * MeSH
- syndromy suchého oka diagnostické zobrazování terapie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
AIMS: To assess the impact of autologous serum eye drops on the level of ocular surface apoptosis in patients with bilateral severe dry eye disease. METHODS: This prospective study was conducted on 10 patients with severe dry eye due to graft versus host disease (group 1) and 6 patients with severe dry eye due to primary Sjögren's syndrome (group 2). Impression cytology specimens from the bulbar conjunctiva were obtained before and after a three-month treatment with 20% autologous serum eye drops applied a maximum of 12 times a day together with regular therapy with artificial tears. The percentage of apoptotic epithelial cells was evaluated immunochemically using anti-active caspase 3 antibody. RESULTS: In group 1, the mean percentage of apoptotic cells was 3.6% before the treatment. The three-month treatment led to a significant decrease to a mean percentage of 1.8% (P = 0.028). The mean percentage of apoptotic conjunctival cells decreased from 5.4% before the treatment to 3.8% in group 2; however, these results did not reach the level of significance. CONCLUSION: Three-month autologous serum treatment led to the improvement of ocular surface apoptosis, especially in the group of patients with severe dry eye due to graft versus host disease. This result supports the very positive effect of autologous serum on the ocular surface in patients suffering from severe dry eye.
- MeSH
- apoptóza fyziologie MeSH
- epitelové buňky fyziologie MeSH
- imunohistochemie MeSH
- konjunktiva fyziologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- nemoc štěpu proti hostiteli patofyziologie MeSH
- oční roztoky aplikace a dávkování MeSH
- prospektivní studie MeSH
- sérum * MeSH
- syndromy suchého oka patofyziologie terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
Cíl: Syndrom suchého oka (SSO) je multifaktoriální onemocnění slz a povrchu oka. V posledních letech se u pacientů trpících SSO používá léčba pomocí očních kapek z autologního séra (AS-ok). Signifikantní zlepšení bylo prokázáno v klinických a laboratorních parametrech i při subjektivním hodnocení pacienty. Předpokládá se, že jedním z klíčových faktorů rozvoje onemocnění je zvýšená osmolarita slz. Cílem naší studie bylo zjistit možnost využití měření osmolarity slz v klinické praxi a ověřit, zda u pacientů s těžkým SSO dojde ke změně osmolarity po tříměsíční aplikaci 20% AS-ok, a zda se hodnoty osmolarity liší u pacientů s těžkou formou SSO a u zdravých jedinců. Metody: Do studie bylo zahrnuto 35 pacientů s těžkým SSO (Schirmerův test < 5 mm/5 min) a 23 zdravých dobrovolníků. U pacientů byly srovnány hodnoty osmolarity slz, Schirmerova testu (ST1), vitálního barvení povrchu oka a hodnoceny subjektivní pocity (dle dotazníku Ocular Surface Disease Index, OSDI) před a po léčbě 20% AS-ok. Dále byly porovnány hodnoty ST1 a osmolarity získané od pacientů se SSO a od zdravých jedinců. Výsledky: U pacientů se SSO došlo po aplikaci AS-ok k signifikantnímu poklesu hodnot vitálního barvení a OSDI (p < 0,0001). Hodnoty ST1 a osmolarity se statisticky významně nezměnily. Hodnoty ST1 byly u zdravých jedinců signifikantně vyšší (p < 0,0001) a hodnoty OSDI signifikantně nižší (p < 0,0001) než hodnoty získané od pacientů. Hodnota osmolarity slz se statisticky nelišila mezi zdravými jedinci (306 mosmol/l) a pacienty se SSO (302 před a 301 mosmol/l po léčbě). Závěr: Stav povrchu oka se u pacientů se SSO po léčbě AS-ok statisticky významně zlepšil na základě vitálního barvení i subjektivního hodnocení pacienty. Zlepšení vitálního barvení a OSDI po léčbě nekorelovalo s hodnotami osmolarity. Významně se nelišily ani hodnoty osmolarity u pacientů a zdravých jedinců. Získané výsledky zpochybňují přínos měření osmolarity pomocí přístroje TearLab Osmolarity System v běžné klinické praxi jak z hlediska diagnostického, tak pro sledování efektu terapie SSO. Klíčová slova: syndrom suchého oka, autologní sérum, osmolarita
Introduction: Dry eye syndrome (DES) is a multifactorial disease of the tears and ocular surface. Recently, treatment with autologous serum eye drops (AS-ed) has been frequently used in these patients. Significant improvement correlates well with clinical, laboratory and subjective findings. It is assumed that one of the key factors in the development of the disease is increased tear osmolarity. The aims of our study were to test tear osmolarity measurements in clinical practice, to examine if osmolarity values differ before and after a 3-month application of 20% AS-ed, and to determine if the values differ between patients with severe DES and healthy individuals. Methods: The study included 35 patients with severe DES (Schirmer test <5 mm/5 min) and 23 healthy volunteers. Tear osmolarity values (TearLab Osmolarity System), the Schirmer test (ST1), vital ocular surface staining and subjective feelings (the OSDI questionnaire) were assessed in patients with DES before and after treatment with 20% AS-ed. Further, the tear osmolarity values were compared between healthy subjects and patients with DES before and after treatment with AS-ed. Results: The values of OSDI and vital staining significantly decreased in patients with DES after the treatment (p < 0.0001). ST1 and tear osmolarity did not change significantly after the treatment. ST1 values in healthy individuals were significantly higher (p < 0.0001) and the OSDI values significantly lower (p < 0.0001) than the results obtained in patients before and after treatment. Tear osmolarity was not statistically different between healthy subjects (306 mosmol/l) and patients with DES (302 and 301 mosmol/l before and after treatment respectively). Conclusion: We demonstrated a positive effect of AS treatment on the ocular surface in patients with DES. However, the osmolarity values did not differ before and after treatment with AS, and they also did not differ significantly between DES patients and healthy individuals. In accordance with other recent studies, our results raise questions concerning the value of the TearLab Osmolarity System for evaluating therapeutic effect and also as a tool for DES diagnosis. Key words: dry eye syndrome, autologous serum, osmolarity
- Klíčová slova
- autologní sérum,
- MeSH
- červeň bengálská diagnostické užití MeSH
- diagnostické techniky oftalmologické přístrojové vybavení statistika a číselné údaje MeSH
- fluorescein diagnostické užití MeSH
- lidé středního věku MeSH
- lidé MeSH
- oční roztoky * terapeutické užití MeSH
- osmolární koncentrace MeSH
- prospektivní studie MeSH
- sérum * MeSH
- slzy fyziologie MeSH
- spokojenost pacientů MeSH
- statistika jako téma MeSH
- syndromy suchého oka * patofyziologie terapie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Klíčová slova
- autologní sérum,
- MeSH
- červeň bengálská diagnostické užití MeSH
- diagnostické techniky oftalmologické přístrojové vybavení statistika a číselné údaje MeSH
- fluorescein diagnostické užití MeSH
- lidé středního věku MeSH
- lidé MeSH
- oční roztoky * terapeutické užití MeSH
- osmolární koncentrace MeSH
- prospektivní studie MeSH
- sérum * MeSH
- slzy fyziologie MeSH
- spokojenost pacientů MeSH
- statistika jako téma MeSH
- syndromy suchého oka * patofyziologie terapie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
AIM: To assess the impact of autologous serum (AS) eye drops on the ocular surface of patients with bilateral severe dry eye and to draw a comparison between the clinical and laboratory examinations and the degree of subjective symptoms before and after serum treatment. MATERIALS AND METHODS: A three-month prospective study was conducted on 17 patients with severe dry eye. AS eye drops were applied a maximum of 12 times a day together with regular therapy. Dry eye status was evaluated by clinical examination (visual acuity, Schirmer test, tear film breakup time, vital staining, tear film debris and meniscus), conjunctival impression cytology (epithelial and goblet cell density, snake-like chromatin, HLA-DR-positive and apoptotic cells) and subjectively by the patients. RESULTS: The application of AS eye drops led to a significant improvement in the Schirmer test (p < 0.01) and tear film debris (p < 0.05). The densities of goblet (p < 0.0001) and epithelial cells (p < 0.05) were significantly increased, indicating a decrease of squamous metaplasia after AS treatment. A significant decrease (p < 0.05) was found in the number of apoptotic, HLA-DR-positive and snake-like chromatin cells on the ocular surface. A significant improvement was found in all evaluated subjective symptoms. Altogether, the clinical results were improved in 77%, the laboratory results in 75% and the subjective feelings in 63% of the eyes. CONCLUSIONS: We found that three-month AS treatment led especially to the improvement of ocular surface dryness and damage of the epithelium. The improvement of dry eye after AS treatment correlated well with the clinical, laboratory and subjective findings. From the patients' subjective point of view, the positive effect of AS decreased with time, but still persisted up to three months after the end of therapy.
- MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- oční roztoky aplikace a dávkování MeSH
- pohárkové buňky účinky léků patologie MeSH
- prospektivní studie MeSH
- rohovkový epitel účinky léků patologie MeSH
- senioři MeSH
- spokojenost pacientů * MeSH
- stupeň závažnosti nemoci MeSH
- syndromy suchého oka farmakoterapie patologie MeSH
- výsledek terapie MeSH
- zraková ostrost MeSH
- zrakové testy MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky MeSH
- práce podpořená grantem MeSH
Po několikaletém trvání diabetu obvykle postupně selhává léčba antidiabetikem první volby metforminem. Studie KOMETA byla prospektivní, multicentrickou, neintervenční epidemiologickou studií u pacientů s diabetes mellitus 2. typu neadekvátně kompenzovaných na terapii metforminem. Studie byla realizována v letech 2010–2011. Studie prokázala signifikantní zlepšení kompenzace diabetu, snížení obvodu pasu, pokles krevního tlaku a úpravu dyslipidemie oproti vstupním hodnotám. Naši lékaři podle této studie volí antidiabetika zodpovědně a používají často léků bez rizika hypoglykemie. Více než 2/3 pacientů přitom dosáhnou kompenzace HbA1c pod 6 % podle IFCC. Hlavním závěrem studie je, že i v běžných podmínkách lze během 9 měsíců dosáhnout komplexní metabolické kompenzace diabetika 2. typu.
After several years of diabetes, the first line anti-diabetic therapy of metformin usually starts failing. The KOMETA study was a prospective, multicentric, non-interventional epidemiological study in patients with type 2 diabetes mellitus inadequately controlled with metformin therapy. The study was conducted between 2010 and 2011. The study demonstrated a significant improvement in diabetes control, reduction of waist circumference, reduction in blood pressure and adjustment of dyslipidemia compared to baseline. Based on this study, our doctors select anti-diabetic drugs responsibly and they often use medicines without the hypoglycaemia risk. Nevertheless, more than two thirds of patients achieve the HbA1c control of under 6% according to IFCC. The main conclusion of this study is that under standard conditions it is possible to achieve full metabolic control of type 2 diabetic patients over the period of nine months.
- MeSH
- diabetes mellitus 2. typu * farmakoterapie MeSH
- dyslipidemie farmakoterapie MeSH
- epidemiologické studie MeSH
- glykovaný hemoglobin * účinky léků MeSH
- HDL-cholesterol MeSH
- hypertenze farmakoterapie MeSH
- hypoglykemika * terapeutické užití MeSH
- index tělesné hmotnosti MeSH
- krevní glukóza MeSH
- LDL-cholesterol MeSH
- lidé středního věku MeSH
- lidé MeSH
- metformin * farmakologie terapeutické užití MeSH
- multicentrické studie jako téma MeSH
- prospektivní studie MeSH
- statistika jako téma MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
