Ability to determine the nontransferrin-bound iron and total iron in the human placenta using high-performance liquid chromatography method

Iron is one of the most important microelements in the human body. It is a component of haemoglobin, which transports oxygen to all cells in the organism. It is also used in the synthesis of myelin, neurotrans-mitters, and DNA and transfers electrons in biochemical reactions. Iron is also responsible for regular development of the foetus’ central nervous system. Furthermore, as a result of Fenton reactions, iron leads to formation of toxic free radicals. The existence of non-transferrin-bound iron (NTBI) and its part desfer-rioxamine-chelatable iron (DCI) can be used to assess this element in the body. The placenta is an organ transition that is formed during pregnancy in the female organism. It has a dense web of blood vessels in which dynamic exchange of blood between mother and foetus takes place. As a result, a fraction of NTBI may be present in the placenta. The main goal of this work was to develop a method for determining total iron and desferrioxamine-chelatable iron in solid tissues - the human placenta.Iron is one of the most important microelements in the human body. It is a component of hemoglobin which transports oxygen to all cells in the organism. It is also used in a synthesis of myelin, neurotransmitters and DNA, and transfers electrons in the biochemical reactions. Iron is also responsible for regular development of a fetus central nervous system. Furthermore, as a result of Fenton reactions, iron leads to formation of toxic free radicals. Existence of non-transferrin-bound iron (NTBI) and its part - desferrioxamine-chelatable iron (DCI) can be used for assess this element in a body. Placenta is an organ transition which is formed during pregnancy in a female organism. It has a dense web of blood vessels where dynamic exchange of blood between mother and fetus takes place. As a result, fraction of NTBI may be present in the placenta. The main goal of this work was to develop a method for determination of total iron and desferrioxamine-chelatable iron in solid tissues - a human placent

Endothelial hemostatic markers in type 2 diabetes and without diabetes in patients with advanced peripheral arterial disease

Wstęp. Cukrzyca typu 2 (DM2) jest ważnym czynnikiem ryzyka w miażdżycy tętnic kończyn dolnych (PAD). Zaburzenia metaboliczne w DM2, jak insulinooporność i hiperglikemia, powodują zwiększoną syntezę reaktywnych tlenowych rodników, które, wywołując proces zapalny i dysfunkcję śródbłonka, prowadzą do aterotromobozy. Uszkodzona ściana naczyniowa uwalnia do krwi śródbłonkowe markery hemostazy. Celem badań było oznaczenie śródbłonkowych markerów hemostazy: czynnika tkankowego (TF), jego inhibitora (TFPI), trombomoduliny (TM), czynnika von Willebranda (vWF), tkankowego aktywatora plazminogenu (t-PA) i inhibitora aktywatorów plazminogenu typu 1 (PAI-1) we krwi chorych z DM2 i bez DM2 u pacjentów z zaawansowaną PAD. Materiał i metody. Śródbłonkowe markery hemostazy oznaczono w osoczu krwi 91 pacjentów z zaawansowaną PAD i DM2, w tym 37 kobiet i 54 mężczyzn w wieku 65,2 ± 9,0 lat. Grupę kontrolną stanowiło 59 pacjentów z zaawansowaną PAD, ale bez DM2. Obie grupy były w podobnym przedziale wieku i płci, z podobnymi czynnikami ryzyka i w większości współistniejącymi chorobami. Jedynie u pacjentów z DM2 częściej występowała dyslipidemia (p < 0,022). Krew pobierano rano na czczo do 3,2% cytrynianu sodu w proporcji 9:1. Śródbłonkowe markery hemostazy oznaczono metodami immunoenzymatycznymi ELISA przy zastosowaniu komercyjnych zestawów firmy American Diagnostica. Wyniki. Stężenia śródbłonkowych markerów: TF, TFPI, TM, vWF, t-PA i PAI-1 w osoczu pacjentów z PAD i DM2 były podobne jak u pacjentów z PAD, ale bez DM2. Natomiast pacjenci z PAD i DM2 oraz z PAD bez DM2 mieli stężenia TF i vWF istotnie statystycznie wyższe (odpowiednio p < 0,05 i p < 0,0001), a TM niższe (p < 0,001) w porównaniu z grupą referencyjną osób zdrowych. Za różnice były odpowiedzialne: starszy wiek, PAD i proces rewaskularyzacji. Stężenia TFPI, t-PA i PAI-1 były natomiast prawie jednakowe w trzech grupach: z DM2, bez DM2 i w grupie referencyjnej. Wniosek. Współistniejąca DM2 z PAD nie zwiększała uwalniania śródbłonkowych markerów hemostazy, co może potwierdzać pogląd o wspólnej etiologii DM2 i PAD polegającej na tak zwanej dysfunkcji śródbłonka naczyń.Background. Type 2 diabetes (DM2) plays an important role as a risk factor for peripheral arterial disease (PAD). Metabolic abnormalities in DM2, as insulin resistance and hyperglycemia cause overproduction of reactive oxygen species, which via inflammation and endothelial dysfunction lead to atherothrombosis. The injured arterial wall releases hemostatic endothelial markers. The aim of the study was to determine: tis­sue factor (TF), tissue factor pathway inhibitor (TFPI), thrombomodulin (TM), von Willebrand factor (vWF), tissue plasminogen activator (t-PA) and plasminogen activators inhibitor type 1 (PAI-1) in type 2 diabetes and without diabetes in patients with advanced PAD. Material and methods. Endothelial markers were examined in 91 patients with PAD and DM2; 37 women and 54 men — aged 65.2 ± 9.0 years. Control group concluded 59 PAD patients without DM2. Both groups of patients were in similar compartments of sex and age with similar risk factors and coexisting diseases. Only in patients with DM2 more frequently appeared dyslipidemia (p < 0.022). Blood was collected in the morning to 3.2% sodium citrate in proportion 9:1. Endothelial hemostatic markers were determined with enzyme immunoassay ELISA using commercial tests of American Diagnostica. Results. The levels of endothelial markers: TF, TFPI, TM, vWF, t-PA and PAI-1 in patients with PAD and DM2 were similar compared to patients without DM2. But patients with DM2 had the concentration of TF and vWF significant higher (p < 0.005, p < 0.0001) and TM lower (p < 0.001) in comparison with reference group of healthy patients. But levels of TFPI, t-PA and PAI-1 were almost equal in all three groups of patients with DM2, without DM2 and of healthy persons. Conclusion. DM2 coexisting in PAD patients did not change the release of endothelial hemostatic markers from arterial wall. This confirms the view of “endothelial dysfunction” as common etiology of both diseases PAD and DM2.

Inflammatory markers in peripheral arterial disease patients after endovascular revascularization with new restenosis

Wstęp. Zapalenie odgrywa kluczową rolę w powstawaniu i rozwoju miażdżycy tętnic, także u pacjentów z miażdżycą tętnic kończyn dolnych (PAD). Zabiegi rewaskularyzacyjne stanowią czynnik ryzyka, który nasila ten proces, niekiedy prowadząc do powstania restenoz.Cel. Oznaczenie stężenia fibrynogenu, białka C-reaktywnego (CRP), interleukin 6 i 10 (IL-6 i IL-10), zasadowego czynnika wzrostu fibroblastów (bFGF) i transformującego czynnika wzrostu (TGF-b1) we krwichorych z PAD po obwodowej, wewnątrznaczyniowej rewaskularyzacji i u pacjentów z restenozami.Materiał i metody. Przebadano 150 pacjentów z PAD, w tym 90 mężczyzn i 60 kobiet w wieku 50–88 (średnio 65,5) lat. Podczas 12-miesięcznej obserwacji po obwodowej, wewnątrznaczyniowej rewaskularyzacji u 38 pacjentów powstały restenozy. Grupa kontrolna dla oznaczanych cytokin składała się z 15 klinicznie zdrowych osób w wieku 60–83 lat bez niedokrwienia kończyn dolnych. Krew pobierano na czczo do 3,2% cytrynianu sodu w proporcji 9:1 (do oznaczenia fibrynogenu i CRP) i 2,6 ml krwi mieszano z EDTA do pomiaru cytokin: IL-6, IL-10, bFBF, TGF-beta1. Cytokiny badano przy użyciu komercyjnych zestawów metodą immunoenzymatyczną, a fibrynogen i CRP za pomocą analizatora koagulologicznego.Wyniki. We krwi pacjentów z PAD po obwodowej rewaskularyzacji stężenie fibrynogenu i CRP było wyższe od normy laboratoryjnej. Podczas 12-miesięcznej obserwacji parametry te nadal wzrastały: CRP prawie dwukrotnie, a fibrynogen tylko o około 10%. Z badanych cytokin tylko IL-10 i bFGF różniły się istotnie statystycznie od grupy kontrolnej. Cytokiny były również wyższe u pacjentów z powstałymi restenozami, mieściły się jednak, z wyjątkiem TGF-beta1, w szerokich normach laboratoryjnych podanych przez producentów testów.Wnioski. 1. U pacjentów z PAD po wewnątrznaczyniowej rewaskularyzacji stężenie CRP i fibrynogenu podczas rocznej obserwacji wyraźnie się zwiększyło.2. U tych pacjentów obserwowano istotną korelację między prozapalnym CRP i przeciwzapalną interleukiną 10.3. Stężenie CRP, fibrynogenu i cytokin: IL-6, bFGF, TGF-b1 i IL-10 zwiększało się także po powstaniu restenozu pacjentów z PAD, potwierdzając udział w tym procesie miernie nasilonego zapalenia.Introduction. Inflammation plays a central role in the pathophysiology and progression of atherosclerosis, also in patients with peripheral arterial disease (PAD). Revascularization is a risk factor which intensifies this process sometimes leading to newly developed restenosis.Aim. To determine the levels of fibrinogen, C-reactive protein (CRP), interleukin 6 (IL-6), interleukin 10 (IL-10), basic fibroblast growth factor (bFGF) and transforming growth factor-beta 1 (TGF-beta1) in the blood of patients with PAD after endovascular revascularization of lower limbs and in patients with new restenosis.Material and methods. The study included 150 patients with PAD (90 men and 60 women), aged 50–88 (mean 65.5) years after successful peripheral angioplasty (PTA) and/or stenting. Within 12 months after revascularization procedure in 38 PAD patients restenosis occurred. Control group for cytokines consisted of 15 clinically healthy subjects without limb ischaemia aged 60–83 years. Fasting blood was drawn in the morning and put into 3.2% sodium citrate in a 9:1 proportion (for determination of fibrinogen and CRP), and 2.6 ml of blood was mixed with EDTA for determination of cytokines: IL-6, IL-10, bFGF and TGF-beta1. All parameters were measured in the plasma of patients with commercial kits for enzyme immunoassay; fibrinogen and CRP levels were measured with coagulometer.Results. In the plasma of patients with PAD after revascularization, the levels of fibrinogen and CRP were significantly higher than in controls. These markers increased within 12 months of observation: CRP level increased almost two-fold and fibrinogen only by about 10%. All examined cytokines in PAD patients with restenosis were higher than in those without restenosis, but the difference was significant only for IL-10 and bFGF. But increased cytokines in patients with restenosis were within the laboratory norms specified by reagent producers except of TGF-beta1.Conclusions.1.In PAD patients after endovascular revascularization the levels of CRP and fibrinogen during one yearobservation significantly increased.2.In these patients, significant correlation was observed between pro-inflammatory CRP and anti-inflam-matory IL-10.3.The concentrations of CRP, fibrinogen and cytokines (IL-6, bFGF, TGF-beta1 and IL-10) were also higher inPAD patients with restenosis indicating the involvement of low-grade inflammation in this process

Retrospective analysis of ocular disorders and frequency of cesarean sections for ocular indications in 2000-2008 – our own experience

Summary 1. Evaluation of frequency of cesarean sections for ocular indications. 2. Analysis of ophthalmological disorders as indications for cesarean section. Material and methods: 4895 cesarean sections were performed (100 due to ocular indications) in the Department of Obstetrics, Female Pathology and Oncological Gynecology, between 2000 and 2008. Medical documentation was analyzed. Results: Among 4895 patients undergoing cesarean sections, 100 (2.04%) presented a written certification from an ophthalmologist suggesting this way of delivery. The frequency of c-sections due to ocular indications continued to increase between 2000-2005 and has been in decline since 2006. The most common ophthalmological disorders included myopia (57%), retinopathy (20%), glaucoma (5%), imminent retinal detachment (4%) and past retinal detachment (3%). In 45% of patients an eye pathology was the only reason for a cesarean section. Conclusion: 1. The frequency of cesarean sections due to ocular reasons in our material was 0.7%- 3.44%, average 2.04%. 2. Since 2006 the number of ocular indications for cesarean section has been decreasing. Nevertheless, it remains to be twice as high as in 2000. 3. The most common eye disorders leading to cesarean section were myopia and retinopathy. 4. In almost half of the patients the decision to conduct a cesarean section was based solely on ophthalmological indications

Polish statement on food allergy in children and adolescents

An adverse food reaction is defined as clinical symptoms occurring in children, adolescents or adults after ingestion of a food or chemical food additives. This reaction does not occur in healthy subjects. In certain individuals is a manifestation of the body hypersensitivity, i.e. qualitatively altered response to the consumed food. The disease symptoms observed after ingestion of the food can be triggered by two pathogenetic mechanisms; this allows adverse food reactions to be divided into allergic and non-allergic food hypersensitivity (food intolerance). Food allergy is defined as an abnormal immune response to ingested food (humoral, cellular or mixed). Non-immunological mechanisms (metabolic, pharmacological, microbiological or other) are responsible for clinical symptoms after food ingestion which occur in non-allergic hypersensitivity (food intolerance). Food allergy is considered a serious health problem in modern society. The prevalence of this disorder is varied and depends, among other factors, on the study population, its age, dietary habits, ethnic differences, and the degree of economic development of a given country. It is estimated that food allergy occurs most often among the youngest children (about 6-8% in infancy); the prevalence is lower among adolescents (approximately 3-4%) and adults (about 1-3%). The most common, age-dependent cause of hypersensitivity, expressed as sensitization or allergic disease (food allergy), are food allergens (trophoallergens). These are glycoproteins of animal or plant origine contained in: cow's milk, chicken egg, soybean, cereals, meat and fish, nuts, fruits, vegetables, molluscs, shellfish and other food products. Some of these allergens can cause cross-reactions, occurring as a result of concurrent hypersensitivity to food, inhaled or contact allergens. The development of an allergic process is a consequence of adverse health effects on the human body of different factors: genetic, environmental and supportive. In people predisposed (genetically) to atopy or allergy, the development of food allergy is determined by four allergic-immunological mechanisms, which were classified and described by Gell-Coombs. It is estimated that in approximately 48-50% of patients, allergic symptoms are caused only by type I reaction, the IgEmediated (immediate) mechanism. In the remaining patients, symptoms of food hypersensitivity are the result of other pathogenetic mechanisms, non-IgE mediated (delayed, late) or mixed (IgE mediated, non-IgE mediated). Clinical symptomatology of food allergy varies individually and depends on the type of food induced pathogenetic mechanism responsible for their occurrence. They relate to the organ or system in which the allergic reaction has occurred (the effector organ). Most commonly the symptoms involve many systems (gastrointestinal tract, skin, respiratory system, other organs), and approximately 10% of patients have isolated symptoms. The time of symptoms onset after eating the causative food is varied and determined by the pathogenetic mechanism of the allergic immune reaction (immediate, delayed or late symptoms). In the youngest patients, the main cause of food reactions is allergy to cow’s milk. In developmental age, the clinical picture of food allergy can change, as reflected in the so-called allergic march, which is the result of anatomical and functional maturation of the effector organs, affected by various harmful allergens (ingested, inhaled, contact allergens and allergic cross-reactions). The diagnosis of food allergy is a complex, long-term and time-consuming process, involving analysis of the allergic history (personal and in the family), a thorough evaluation of clinical signs, as well as correctly planned allergic and immune tests. The underlying cause of diagnostic difficulties in food allergy is the lack of a single universal laboratory test to identify both IgE-mediated and non-IgE mediated as well as mixed pathogenetic mechanisms of allergic reactions triggered by harmful food allergens. In food allergy diagnostics is only possible to identify an IgE-mediated allergic process (skin prick tests with food allergens, levels of specific IgE antibodies to food allergens). This allows one to confirm the diagnosis in patients whose symptoms are triggered in this pathogenetic mechanism (about 50% of patients). The method allowing one to conclude on the presence or absence of food hypersensitivity and its cause is a food challenge test (open, blinded, placebo-controlled). The occurrence of clinical symptoms after the administration of food allergen confirms the cause of food allergy (positive test) whereas the time elapsing between the triggering dose ingestion and the occurrence of clinical symptoms indicate the pathogenetic mechanisms of food allergy (immediate, delayed, late). The mainstay of causal treatment is temporary removal of harmful food from the patient’s diet, with the introduction of substitute ingredients with the nutritional value equivalent to the eliminated food. The duration of dietary treatment should be determined individually, and the measures of the effectiveness of the therapeutic elimination diet should include the absence or relief of allergic symptoms as well as normal physical and psychomotor development of the treated child. A variant alternative for dietary treatment of food allergy is specific induction of food tolerance by intended contact of the patient with the native or thermally processed harmful allergen (oral immunotherapy). This method has been used in the treatment of IgE-mediated allergy (to cow's milk protein, egg protein, peanut allergens). The obtained effect of tolerance is usually temporary. In order to avoid unnecessary prolongation of treatment in a child treated with an elimination diet, it is recommended to perform a food challenge test at least once a year. This test allows one to assess the body's current ability to acquire immune or clinical tolerance. A negative result of the test makes it possible to return to a normal diet, whereas a positive test is an indication for continued dietary treatment (persistent food allergy). Approximately 80% of children diagnosed with food allergy in infancy "grow out" of the disease before the age of 4-5 years. In children with non-IgE mediated food allergy the acquisition of food tolerance is faster and occurs in a higher percentage of treated patients compared to children with IgE-mediated food allergy. Pharmacological treatment is a necessary adjunct to dietary treatment in food allergy. It is used to control the rapidly increasing allergic symptoms (temporarily) or to achieve remission and to prevent relapses (long-term treatment). Preventive measures (primary prevention of allergies) are recommended for children born in a "high risk" group for the disease. These are comprehensive measures aimed at preventing sensitization of the body (an appropriate way of feeding the child, avoiding exposure to some allergens and adverse environmental factors). First of all, the infants should be breast-fed during the first 4-6 months of life, and solid foods (non milk products, including those containing gluten) should be introduced no earlier than 4 months of age, but no later than 6 months of age. An elimination diet is not recommended for pregnant women (prevention of intrauterine sensitization of the fetus and unborn child). The merits of introducing an elimination diet in mothers of exclusively breast-fed infants, when the child responds with allergic symptoms to the specific diet of the mother, are disputable. Secondary prevention focuses on preventing the recurrence of already diagnosed allergic disease; tertiary prevention is the fight against organ disability resulting from the chronicity and recurrences of an allergic disease process. Food allergy can adversely affect the physical development and the psycho-emotional condition of a sick child, and significantly interfere with his social contacts with peers. A long-term disease process, recurrence of clinical symptoms, and difficult course of elimination diet therapy are factors that impair the quality of life of a sick child and his family. The economic costs generated by food allergies affect both the patient's family budget (in the household), and the overall financial resources allocated to health care (at the state level). The adverse socio-economic effects of food allergy can be reduced by educational activities in the patient’s environment and dissemination of knowledge about the disease in the society

Differential inflammatory microRNA and cytokine expression in pulmonary sarcoidosis

Sarcoidosis is a granulomatous disease of unknown etiology. The disease has an important inflammatory and immune component; however, its immunopathogenesis is not completely understood. Recently, the role of microRNAs (miRNAs), the small non-coding RNAs, has attracted attention as both being involved in pathogenesis and serving as disease markers. Accordingly, changes in the expression of some miRNAs have been also associated with different autoimmune pathologies. However, not much is known about the role of miRNAs in sarcoidosis. Therefore, the aim of this study was to compare the level of expression of selected miRNAs in healthy individuals and patients with sarcoidosis. We detected significantly increased level of miR-34a in peripheral blood mononuclear cells isolated from sarcoidosis patients. Moreover, significantly up-regulated levels of interferon (IFN)-γ, IFN-γ inducible protein (IP-10) and vascular endothelial growth factor were detected in sera of patients when compared to healthy subjects. Our results add to a known inflammatory component in sarcoidosis. Changes in the levels of miR-34a may suggest its involvement in the pathology of this disease