2,3,7,8-Tetrachlorodibenzo-p-dioxin alters steroid secretion but does not affect cell viability and the incidence of apoptosis in porcine luteinised granulosa cells

The compound 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a by-product of human industrial activity, was found to affect ovarian steroidogenesis in animals, but the mechanism of its action is still unclear. The aims of the study were to examine the effect of TCDD on (1) progesterone (P4) and oestradiol (E2) production by granulosa cells isolated from medium (3–6 mm) and preovulatory (≥ 8 mm) porcine follicles, (2) the viability of the cells, and (3) the incidence of apoptosis. Porcine granulosa cells were cultured (48 h) with or without TCDD (100 pM, 100 nM). Steroid hormone concentrations in the medium were determined by radioimmunoassay. The viability of granulosa cells was tested spectrophotometrically (alamarBlue™ assay). Apoptosis was evaluated by flow cytometry using Annexin V and by TUNEL assay. The higher dose of TCDD (100 nM) significantly inhibited P4 and stimulated E2 production by luteinised granulosa cells isolated from medium follicles. The lower dose of TCDD (100 pM) significantly stimulated P4 and inhibited E2 secretion by the cells isolated from preovulatory follicles. None of the two TCDD doses affected cell viability or induced apoptosis in granulosa cells. In conclusion, TCDD directly affected steroid production by granulosa cells obtained from mature pigs, but the effect of TCDD was not due to its cytotoxicity

Apoptotic cell death in the porcine endometrium during the oestrous cycle

It has been reported that apoptosis plays an essential role in controlling the physiological cell kinetics in the human and rodent endometrium but this type of death has never been studied in the porcine endometrium. The aim of this study was to investigate the apoptotic cell death in the porcine endometrium during the middle (Days 9–11) and late (Day 13) luteal phase, during the luteolysis (Day 15) and early follicular phase (Days 17–19) of the oestrous cycle. Apoptotic cells were identified by in situ DNA 3′-end labelling method. it was revealed that the greatest number of apoptotic cells in the luminal and glandular epithelium was found on Days 17–19 and on Day 15 of the oestrous cycle, respectively. in the stroma, the greatest number of these cells was found on Days 9–11. Our data have shown that in the porcine endometrium, both epithelial and stromal cells undergo apoptosis and that the number of apoptotic cells varies depending on the phase of the oestrous cycle

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

Pielęgniarstwo na rzecz milenijnych celów rozwoju. Cz. 2

Praca recenzowana / Peer-reviewed paperZ przyjemnością oddajemy w Państwa ręce drugi tom monografii wydanej z okazji XII Kongresu Pielęgniarek Polskich w Krakowie, w którym udział wzięło blisko 500 pielęgniarek, zaprezentowano 160 prac w 9 sesjach. Ta kolejna już publikacja świadczy o dynamicznym rozwoju badań naukowych w pielęgniarstwie. Dowodzi, że pielęgniarstwo to nie tylko zawód, ale i interdyscyplinarna nauka

ORIGINAL PAPER Local

vascular pathway for progesterone transfer to the brain after nasal administration in gilt

Carbon Monoxide (CO) as a Retinal Regulator of Heme Oxygenases -1, and -2 (HO’s) Expression

Carbon monoxide (CO) has been proposed as a chemical light signal and neural system modulator via heme oxygenases -1 and -2 (HO-1 and HO-2). Many papers have proven the CO-HO circuit to be important for such physiological pathways as the molecular biological clock and the GnRH axis, but also in such pathological occurrences as ischemic injuries, or inflammation as a regenerative and neuroprotective factor. In this in vivo experiment, we used three groups of pigs: control—housed in natural conditions without any procedures; without CO—adapted and kept in constant darkness, infused with blank plasma; and with CO—adapted and kept in constant darkness infused with CO-enriched plasma. After the experiments, each animal was slaughtered and its eyes were collected for further analysis. Quantitative PCR and Western blot analysis were performed to show statistical differences in the expressions between the experimental groups. Our data revealed that exogenous CO is regulator of mRNA transcription for HO-1 and HO-2 and PCNA. Moreover, the mRNA abundance of analyzed factors in the experimental group after CO elevation revealed a restored gene-expression level similar to the control group, which we had observed in the group’s restored protein level after CO elevation. In conclusion, exogenous CO regulates HO’s and PCNA gene expression on transcriptional and translational levels in a similar way as a light cue

Expression of Transforming Growth Factor Beta Isoforms in Canine Endometrium with Cystic Endometrial Hyperplasia–Pyometra Complex

Cystic endometrial hyperplasia (CEH) and pyometra are the most frequently diagnosed uterine diseases affecting bitches of different ages. Transforming growth factor beta (TGF-β) has been classified in females as a potential regulator of many endometrial changes during the estrous cycle or may be involved in pathological disorders. The aim of this study was to determine the expression of TGF-β1, -β2 and -β3 in the endometrium of bitches suffering from CEH or a CEH–pyometra complex compared to clinically healthy females (control group; CG). A significantly increased level of TGF-β1 mRNA expression was observed in the endometrium with CEH–pyometra compared to CEH and CG. Protein production of TGF-β1 was identified only in the endometrium of bitches with CEH–pyometra. An increase in TGF-β3 mRNA expression was observed in all the studied groups compared to CG. The expression of TGF-β2 mRNA was significantly higher in CEH and lower in CEH–pyometra uteri. The results indicate the presence of TGF-β cytokines in canine endometrial tissues affected by proliferative and degenerative changes. However, among all TGF-β isoforms, TGF-β1 could potentially be a key factor involved in the regulation of the endometrium in bitches with CEH–pyometra complex

Złożona translokacja wariantowa t(9;22;6;17;1) w przebiegu leczenia przypadku przewlekłej białaczki szpikowej.

The presence of the Philadelphia chromosome (Ph) in chronic myelogenous leukaemia (CML) is a specific cytogenetic change resulting from a reciprocal translocation between chromosomes 9 and 22. In 5–10% of newly diagnosed cases there are variant translocations (vPh) involving more chromosomes. This paper presents the case of a CML patient with a complex variant translocation involving chromosomes 1, 6, 9, 17 and 22. A molecular analysis did not reveal any muta­tion in the kinase domain of BCR-ABL1 gene or the mutation of TP53 gene. After the first-line treatment with imatinib no cytogenetic or molecular response was obtained. The change of treatment to dasatinib resulted in a minimal cytogenetic response (minCyR) followed by a minor cytogenetic response (mCyR). The application of nilotinib in the third-line treatment resulted in a complete molecular response (CMolR) and therapy success. The likely reason for the failure of the first- and second-line treatment was the loss of a fragment of the 17p13 region as a result of a variant translocation. The change can be a functional equivalent of the loss of one copy of TP53. The analysis of presented case confirms the significance of the detailed evaluation of the composition of vPh complex variant translocations as well as importance of combination cytogenetic and molecular diagnostics in CML treatment monitoring. It makes possible to adequate diagnose higher-risk patients and apply effective treatment strategies if an aberration is identified.Obecność chromosomu Philadelphia (Ph) w przewlekłej białaczce szpikowej (PBSz) jest swoistą zmianą cytogenetyczną wynikającą z wzajemnej translokacji pomiędzy chromosomami 9 i 22. W 5-10% nowo diagnozowanych przypadków dochodzi do translokacji wariantowych (vPh), które angażują więcej chromosomów. W pracy przedstawiono przypadek 52-letniego mężczyzny ze stwierdzoną w badaniu cytogenetycznym i molekularnym przewlekłą białaczką szpikową ze złożoną translokacją pomiędzy chromosomami: 1, 6, 9, 17 i 22. Analiza molekularna nie wykazała obecności mutacji w domenie kinazowej genu fuzyjnego BCR-ABL1, ani mutacji genu TP53. Po zastosowaniu leczenia imatynibem w I linii, stwierdzono brak odpowiedzi cytogenetycznej oraz molekularnej. Zmiana leczenia na dazatynib spowodowała uzyskanie minimalnej (minCyR), a następnie mniejszej odpowiedzi cytogenetycznej (mCyR). Po zastosowaniu nilotynibu w III linii leczenia uzyskano całkowitą odpowiedź molekularną (CMolR) i powodzenie terapii. Prawdopodobnie przyczyną niepowodzeń I i II linii leczenia była utrata fragmentu obszaru 17p13 w rezultacie translokacji wariantowej. Zmiana ta jest funkcjonalnie równoważna utracie jednej kopii genu TP53. Analiza przedstawionego przypadku potwierdza, jak istotna jest szczegółowa ocena składu złożonych translokacji wariantowych vPh+, a także wspólna diagnostyka cytogenetyczna i molekularna oraz monitorowanie leczenia przebiegu PBSz. Pozwala to na ujawnienie pacjentów podwyższonego ryzyka, a identyfikacja zaburzenia umożliwia zastosowanie skutecznych strategii leczenia. The presence of the Philadelphia chromosome (Ph) in chronic myeloid leukemia (CML) is a specific cytogenetic change resulting from reciprocal translocation between chromosomes 9 and 22. Variant translocations (vPh) involving more chromosomes occur in 5-10% of newly diagnosed cases. In this study we present the case of a 52-year-old man with vPh in complex translocation between five chromosomes: 1, 6, 9, 17 and 22, diagnosed by cytogenetic and molecular studies. Molecular analysis showed no mutation in the BCR-ABL1 kinase domain of the fusion gene and no mutation of the TP53 gene. Patient demonstrated no cytogenetic and molecular response to standard-dose imatinib in the first-line treatment. Dasatinib therapy, after imatinib failure, induced minimal cytogenetic response (minCyR) followed by minor cytogenetic response (mCyR). Complete molecular response (CMolR) was achieved on the third-line nilotinib therapy. The first and second-line treatment failure could be caused by the loss of a fragment within the 17p13 region, which can be functionally equivalent to the deletion of one copy of TP53 gene. The analysis of the presented case confirms the importance of a detailed assessment of the composition of vPh complex variant translocations, as well as combined cytogenetic and molecular diagnostics in monitoring of the treatment of CML. This approach allows detection of high-risk patients and the identification of the abnormalities enables the use of the most effective treatment strategies

Rzadki przypadek złożonej translokacji wariantowej t(9;22;6;17;1) w przewlekłej białaczce szpikowej

Obecność chromosomu Filadelfia (Philadelphia – Ph) w przewlekłej białaczce szpikowej (PBSz) jest swoistą zmianą cy­togenetyczną wynikającą z wzajemnej translokacji pomiędzy chromosomami 9 i 22. W 5–10% nowo diagnozowanych przypadków dochodzi do translokacji wariantowych (vPh), które angażują więcej chromosomów. W pracy przedsta­wiono przypadek pacjenta z PBSz ze złożoną translokacją wariantową pomiędzy chromosomami: 1, 6, 9, 17 i 22. Analiza molekularna nie wykazała obecności mutacji w domenie kinazowej genu fuzyjnego BCR-ABL1, ani mutacji genu TP53. Po zastosowaniu leczenia imatynibem w 1. linii stwierdzono brak odpowiedzi cytogenetycznej oraz molekularnej. Zmiana leczenia na dazatynib spowodowała uzyskanie minimalnej (minCyR), a następnie mniejszej odpowiedzi cytogenetycznej (mCyR). Po zastosowaniu nilotynibu w 3. linii leczenia uzyskano całkowitą odpowiedź molekularną (CMolR) i powodzenie terapii. Prawdopodobnie przyczyną niepowodzeń 1. i 2. linii leczenia była utrata fragmentu obszaru 17p13 w rezultacie translokacji wariantowej. Zmiana ta jest funkcjonalnie równoważna utracie jednej kopii genu TP53. Analiza przedstawio­nego przypadku potwierdza, jak istotna jest szczegółowa ocena składu złożonych translokacji wariantowych vPh, a także wspólna diagnostyka cytogenetyczna i molekularna w monitorowaniu przebiegu leczenia PBSz. Pozwala to na rozpoznanie pacjentów podwyższonego ryzyka, a identyfikacja zaburzenia umożliwia zastosowanie skutecznych strategii leczenia