Insight into "nuclear-cytoplasmic shuttling" as a developmental and differentiational capability of cells in primary culture models.

Shuttling proteins are molecules that can facilitate transport through the nuclear envelope. A very large number of proteins are involved in this process that includes nuclear pore buildup, signal, receptor and enzyme proteins. There are many examples of proteins whose biological activity depends on nucleocytoplasmic transport. Very often they are largely responsible for the proper occurrence of cell division, maturation, development and differentiation. Thanks to the well mastered methods of in vitro cell culture, it is possible to trace the levels of protein expression and their distribution in cells. Advanced molecular techniques allow for precise determination of their displacement in time. Several studies are still being carried out, using primary cultures, to identify the factors that determine the maturation, development and differentiation of cells. In understanding of the detailed mechanisms controlling cell life, the key is not the level of expression of a specific protein, but its distribution in individual cellular compartments.

Balance between epithelial and stromal marker expression and distribution in primary culture model of porcine endometrium during real-time cell proliferation..

The similarity between humans and pigs, when it comes to tissue morphology, makes Sus scrofa not only a good research model, but also a potential source of cells for tissue engineering. Cell samples obtained from the pig donor, could be influenced in vitro, in order to become a source of tissue material for xenotransplantation, reconstructive and regenerative medicine. Significant amounts of data point to especially major similarities in pig and human reproductive systems. Because of that, particular scientific focus is centered on research concerning porcine COCs, theca and granulosa cells in primary cultures. One of the aspects of the reproductive process, that is still largely undiscovered, is the interaction between preimplantation blastocyst and maternal uterine tissues. In this study, we used molecular analysis techniques, such as RT-qPCR and immunocytochemistry, to analyze the expression and distribution of cytokeratin 18 and panCytokeratins 8, 18 and 19 and vimentin in porcine luminal endometrial epithelial cells, coupled with analysis of their behavior in RTCA. The results have confirmed the presence of epithelial, as well as stromal cell markers in the cells, varying in levels at different stages of culture. They have also given insight into the modes of proliferation and differentiation of studied cells in in vitro culture, as well as providing additional proof for the possible mesenchymal transdifferentiation of epithelial cells.

Enzyme linked receptor protein signaling pathway is one of the ontology groups that are highly up-regulated in porcine oocytes before in vitro maturation.

Before being able to fully participate in the processes associated with its function as a female gamete, the oocyte needs to undergo a range of changes to achieve its mature form. These morphological, biochemical and metabolomic processes are induced by the somatic tissues surrounding the oocyte, through the expression of specific transcription and growth factors. The maturation of the oocyte is highly important for the proceedings that lead to successful fertilization, early embryonic development and implantation. Domestic pigs were used as models for our study, with the cumulus-oocyte complexes obtained from the ovaries that were recovered at slaughter. After shedding of the cumulus, oocytes were assessed with BCB test, with the viable ones chosen to undergo in vitro maturation. With the use of expression microarrays, we analyzed gene expression before and after IVM and detected major changes in both genes that were proven to be associated with oocyte maturation before (FOS, VEGFA, CHRDL1, TGFBR3, FST, INSR, ID1, TXNIP, SMAD4, MAP3K1, EIF2AK3 and KIT) and genes not previously linked with reproduction associated processes (MYO1E, PHIP, KLF10 and SHOC2). All the genes were briefly described, with consideration of possible involvement of the newly discovered elements of the transcriptome in the process of oocyte maturation.

ADAM protein family - its role in tumorigenesis, mechanisms of chemoresistance and potential as diagnostic and prognostic factors.

ADAMs are a family of transmembrane proteins described for the first time in the 1990's. ADAMs is an abbreviation of "A Disintegrin and Metallo-proteinases". Their earliest known role was involvement in gamete fusion, and their adhesion properties in intercellular interactions also suggested involvement in tumor biology. Further research emphasized the importance of ADAM proteins in the regulation of neoplastic processes due to their influence on adhesion, cell migration, proteolysis and cell signaling. Variable ADAM expression in cancer and normal tissue was the basis for considering these proteins as diagnostic markers. Recent numerous studies have been published suggesting the prognostic value of this protein family members. The ADAMs transmembrane proteins regulate processes associated with carcinogenesis and neoplastic progression, including immune response evasion, growth induction and metastasis. Proteolysis and shedding of membrane proteins and binding integrins by ADAMs lead to the activation of numerous growth factors, changes in the extracellular matrix, adhesion proteins and angiogenesis. ADAMs potential as prognostic and diagnostic markers in cancer treatment is a particularly interesting issue and has great practical significance. There are many new studies concerning ADAMs' roles in carcinogenesis, but there are no recent reviews of the latest developments in this field. The aim of this systematic review is to analyze the results of studies published on ADAMs in the last 5 years, to present their roles in neoplasm pathogenesis and their potential utility in clinical oncology.

Gastroesophageal Reflux Disease in Children with Interstitial Lung Disease.

Gastroesophageal reflux disease is common in adult patients with interstitial lung disease. However, no data currently exist regarding the prevalence and characteristics of the disease in pediatric patients with interstitial lung disease. The aim of the present study was to prospectively assess the incidence of gastroesophageal reflux disease and characterize its features in children with interstitial lung disease. Gastroesophageal reflux disease was established based on 24 h pH-impedance monitoring (MII-pH). Gastroesophageal reflux episodes (GERs) were classified according to widely recognized criteria as acid, weakly acid, weakly alkaline, or proximal. Eighteen consecutive patients (15 boys, aged 0.2-11.6 years) were enrolled in the study. Gastroesophageal reflux disease was diagnosed in a half (9/18) of children. A thousand GERs were detected by MII-pH (median 53.5; IQR 39.0-75.5). Of these, 585 (58.5 %) episodes were acidic, 407 (40.7 %) were weakly acidic, and eight (0.8 %) were weakly alkaline. There were 637 (63.7 %) proximal GERs. The patients in whom gastroesophageal reflux disease was diagnosed had a significantly higher number of proximal and total GERs. We conclude that the prevalence of gastroesophageal reflux disease in children with interstitial lung disease is high; thus, the disease should be considered regardless of presenting clinical symptoms. A high frequency of non-acid and proximal GERs makes the MII-pH method a preferable choice for the detection of reflux episodes in this patient population.

Evaluation of laryngopharyngeal reflux in pediatric patients with asthma using a new technique of pharyngeal pH-monitoring.

There is a debate about the association between asthma and gastroesophageal and/or laryngopharyngeal reflux (LPR). Pharyngeal pH-monitoring is a new technique that allows a physician to assess whether reflux passes the upper esophageal sphincter barrier. The aim of the study was to assess the prevalence of LPR in children with difficult-to-treat asthma. The present study was an open, prospective one. A total of 21 subjects of the mean age 12.7 years were enrolled in the study. All children were asked to fill out a Reflux Symptoms Index questionnaire and a 24-h pharyngeal pH monitoring was performed, using the Dx-pH Measurement System. The LPR was diagnosed in 13 (61.9%) children. There was a positive correlation between LPR diagnosis and the degree of asthma control. The LPR was more frequent in children treated with a higher than lower doses of fluticasone (p = 0.019, OR = 17.3) and in those using montelukast compared with non-users (p = 0.008, OR = 19.0). The mean Reflux Symptoms Index score was almost twice greater in children with LPR than in those without it (13.2 vs. 6.8, respectively, p = 0.003). We conclude that the prevalence of laryngopharyngeal reflux in children with difficult-to-treat asthma is substantial.

The use of anti-asthmatic inhalation therapy and the risk of dental caries in a group of Polish children: a prospective study.

AIM: Assessment of the effect of inhaled therapy with anti-asthmatic medications on the development of caries in children between 3 and 17 years old. METHODS: Study design: The study involved 208 patients, 114 in the study group and 94 in the control group. Data on general health condition of patients, use of medications, dietary habits and oral hygiene were obtained. Information on the onset of asthma, its severity and type, doses and method of administration of asthma medications were obtained from the available medical records by the paediatrician. Dental examination was performed using the DMFT/dmft and DMFS/dmfs index, ICDAS II classification and Nyvad's criteria for noncavitated white spot lesion differentiation. Additionally, an assessment of salivary S. mutans (SM) and Lactobacillus spp. (LA) was carried out using the CRT Bacteria (Ivoclar Vivadent) test. CONCLUSION: There is still no clear consensus among the authors as to whether asthma and medications used in its treatment increase the risk of caries. Nowadays, despite better access to various sources of information, there is still a need for increased awareness in patients and medical doctors on the dental care and caries prevention in patients with asthma.

Basophil activation test based on the expression of CD203c in the diagnostics of cow milk allergy in children.

BACKGROUND: Detection of allergen-induced basophil activation by flow cytometry has been shown to be a useful tool for allergy diagnosis. OBJECTIVE: The aim of this study was to assess the potential of this technique for the diagnosis of pediatric cow milk allergy. MATERIAL AND METHODS: The quantification of total and specific IgE and basophil activation test were performed to evaluate cow milk allergic (n = 9), and non-allergic children (n = 15). RESULTS: Allergen-induced basophil activation was detected as a CD203c up-regulation. The expression of CD203c antigen on basophils was measured with flow cytometry. The antigen CD203c was detected on 15.4 ±10.2% basophils from allergic children after incubation with specific allergens in concentration 1:10, whereas in the control group there were 3.0 ±1.5% of basophils positive for this molecule (P<0.05). Stimulation with allergen diluted 1:500 resulted in activation of 15.3 ±11.2% of basophils in allergic children and 3.8 ±2.3% of cells in the control group (P<0.05). Positive results of an allergenicity test (above the cut- off level of 10%) were obtained in 7 out of the 9 allergic children. In 5 cases, the cut-off level was reached with both dilution of allergens (1:10 and 1:500). In 1 patient, positive stimulation was observed after stimulation with allergen diluted 1:10 and in another case only 1:500 resulted in stimulation of more then 10% of basophils. In no child of the control group, stimulation above 10% was noted. CONCLUSIONS: This study demonstrates that the analysis of allergen-induced CD203c up-regulation with flow cytometry is a reliable tool for the diagnosis of cow milk allergy in pediatric patients, with sensitivity similar to routine diagnostic tests and a higher specificity.

Inverted and horizontal impacted third molars in an Early Modern skull from Wroclaw, Poland: a case report.

BACKGROUND: An impacted tooth is one of the most commonly occurring dental anomalies, although some types of impaction (i.e. inverted angulation) may be considered rare finding. There are many hypotheses regarding impaction aetiology. One of the most popular hypotheses suggested that this condition may result from insufficient space in retromolar space, other: improper angulation of tooth bud, malposition of the tooth germ or hereditary factors, insufficient interproximal attrition, ectopy or dysfunction of genes necessary for proper tooth eruption. This study aims to present the odontological and paleopathological assessment of the impacted molars observed within the skull excavated from an early modern cemetery in Wroclaw. MATERIALS AND METHODS: The skull used in the study was complete and in a good state of preservation. It belonged to an adult individual whose body was buried at the former Salvator Cemetery (currently Czysty Square). The individual's dentition was almost completely lost antemortem. Only second molars preserved within the maxillae (bilaterally) and the mandible was almost edentulous as well. The morphometric traits have been taken according to standards established by R. Martin. Macroscopic observations were supported by X-rays and computed tomography imaging. RESULTS: The age at death was estimated at 20-35 years. Comparison of the metric characteristics of skull with the reference material reveals that it is much smaller than the average female skull from this series. Morphometric indices calculated for both splanchocranium and neurocranium allow defining the skull and jaw as short, which could be an important factor involved in the teeth impaction. CONCLUSIONS: Atypical impaction of the third molars could result from small size of skull and could have significantly deteriorated the quality of life of the individual.

[Is it possible to predict the result of the bronchial provocation test in children with asthma?]. / Czy mozna przewidziec wynik testu prowokacyjnego oskrzeli u dzieci chorych na astme?

Bronchial hyperreactivity (BHR) to different allergic and non-allergic stimuli is characteristic feature of asthma. Sometimes however it is not possible to perform bronchial provocation test (BPT) assessing reactivity. It was interesting for us if the result of BPT can be predicted on the base of routine lung tests. The aim of the study is evaluation of the relationship between BPT results and baseline lung function tests assessing small bronchi obstruction in children suffering from asthma. Investigated group comprised 139 children aged 7 to 17 years, with episodic, mild or moderate asthma. During bronchial challenge lung function was assessed on the base of spirography and maximal flows at 50% and 25% of forced vital capacity (MEF50 and MEF25) and other indices as surface under end-half of flow-volume curve and mean flow times T50 and T25. The study results confirmed good correlation between BPT result and baseline lung function. Those children which had worse initial lung tests had more pronounced bronchial hyperreactivity. This relationship was the closest in the group of children with small bronchi obstruction. Analysis of correlation showed highly significant relationship between baseline lung function tests and degree of bronchial reactivity. The highest significance was observed for MEF50 and MEF25. We conclude that small bronchi test disturbances in children with asthma could predict with high probability results of bronchial challenge.

[Reactivity of the bronchi as a function of their initial state in children with asthma]. / Reaktywnosc oskrzeli jako funkcja ich stanu wyjsciowego u dzieci chorych na astme.

The aim of the study was to assess the effect of peripheral bronchial base-line status on the result of the bronchial provocation test. The study was carried out on a group of 65 children with bronchial asthma. Bronchial reactivity was assessed utilizing the carbachol provocation test. Higher reactivity was seen in children with initially lower indices of peripheral bronchial flow (MEF50, MEF25) (PC20 = 3,789 +/- 0.014 mg/ml) in comparison with children with normal indices (PC20 = 74,608 +/- 0,038 mg/ml) (p < 0.001). A linear correlation was found between the results of bronchial provocation (PC) and initial standardized small airway flow values (MEF50 - r = 0.56, p < 0.001; MEF25 - r = 0.49, p < 0.001). The correlation between bronchial provocation and initial FEV1/VC was weaker (r = 0.39 p < 0.01). The authors conclude that demonstration of decreased flow in peripheral, small airways utilizing MEF values enable to predict a positive result of bronchial provocation tests.

[Evaluation of the effect of gluten-free diet on nutritional status of children with florid celiac disease]. / Ocena wplywu diety bezglutenowej na stan odzywienia dzieci z celiakia "kwitnaca".

A dynamics of Nutritional Status of 55 children with recently diagnosed coeliac disease was assessed during the treatment with gluten free diet. No deterioration of the nutritional status was seen in 34.1% of the investigated children (time 0) whereas different degree of nutritional deficiency was noted in the remaining 75.9% of children including 10% of children with severe deficiency. After a 2-year therapy with gluten-free diet a deteriorated nutritional status was only found in 8.6% of children. In 27 children with gluten free diet for nearly 3 years, with maintenance of histological remission, the percentage of children with nutritional deficiency was 7.2%. The fact of normal Nutritional Status in more than 30% of the investigated coeliac children at the time of the initial diagnosis seems to be an effect of early and proper initial diagnosis.

[The effect of 6-month provocation with gluten on the height and weight of children with celiac disease]. / Wplyw 6-miesiecznej prowokacji glutenem na wysokosc i mase ciala u dzieci z celiakia.

Gluten challenge was performed in 22 coeliac children in the period of histological remission. Gluten in dose 0.5 g/kg was administered for 6 months. Before the Provocation Test over 60% of children have had body weight over 50 c for normal children in Warsaw. Standardized mean body weight (+ 0.13 +/- 1.1) and height (+ 0.04 +/- 1.05) of the investigated coeliac children did not differ significantly from the values of normal Warsaw population (p greater than 0.005). A 6 month provocation with gluten leading to the histological relapse did not affect body weight and height of tested children; over 50% have had body weight and height over 50 c. Mean standardized body weight (- 0.002 +/- 1.0) and height height (-0.07 +/- 0.91) did not differ significantly from the normal values (p greater than 0.05) and corresponding values before the provocation (p greater than 0.05). It seems that Provocation Test with gluten carried out in the above way does not inhibit physical development of children with coeliac disease.

Inflammatory cytokines in exhaled breath condensate in children with inflammatory bowel diseases.

BACKGROUND: Although inflammatory bowel diseases (IBD) affect mainly the gastrointestinal tract, the extra-intestinal manifestations are not uncommon. Different diagnostic methods have been applied to assess pulmonary involvement in patients with IBD, but majority of these methods show significant limitations in children. The aim of our study was to evaluate the usefulness of exhaled breath condensate (EBC) measurements of pro-inflammatory cytokines in children with IBD. MATERIAL AND METHODS: Twenty-two children with Crohn's disease (CD) (mean age 13.8 ± 3.3 years), 25 with ulcerative colitis (UC) (mean age 14.1 ± 3.3 years) and 37 healthy volunteers (mean age 13.9 ± 3.6 years) were studied. IBD activity was assessed using appropriate scoring systems. None of the patients had signs or symptoms of pulmonary disease. Exhaled breath condensate was collected and EBC concentration of interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and interleukin 8 (IL-8) was measured. RESULTS: The concentrations of all the assessed cytokines were significantly higher in the study group as compared to controls. A negative correlation between IL-1ß and CD activity index was found. There were no significant relationships between TNF-α, IL-6, or IL-8 level and CD activity index as well as between IL-1ß, TNF-α, IL-6, IL-8 and UC activity index. No significant correlation between the concentration of IL-1ß, TNF-α, IL-6, IL-8, and IBD duration or treatment duration was found. CONCLUSIONS: Elevated concentration of pro-inflammatory cytokines in EBC in children with IBD may suggest the presence of asymptomatic inflammation in the lower airways.

Omalizumab in children with inadequately controlled severe allergic (IgE-mediated) asthma.

BACKGROUND: Many children with severe persistent allergic (IgE-mediated) asthma remain inadequately controlled despite treatment with high-dose inhaled corticosteroids (ICS) plus a long-acting beta(2)-agonist (LABA). RESEARCH AND DESIGN METHODS: This pre-specified analysis of a randomized, double-blind, placebo-controlled trial evaluated the efficacy and safety of omalizumab in children (6-<12 years) with perennial allergen sensitivity, and history of asthma exacerbations and symptoms despite treatment with ICS (fluticasone >or=500 microg x day(-1) or equivalent) plus a LABA. Patients received omalizumab (75-375 mg once or twice a month by subcutaneous injection, as determined from dosing tables) or placebo over 52 weeks (24-week fixed-steroid then 28-week adjustable-steroid phases). RESULTS: Out of 246 randomized patients (omalizumab, n = 166; placebo, n = 80), efficacy was analysed in 235 (omalizumab, n = 159; placebo, n = 76). Over the 24-week fixed-steroid phase, omalizumab reduced the rate of clinically significant asthma exacerbations (worsening symptoms requiring doubling of baseline ICS dose and/or systemic steroids) by 34% versus placebo (0.42 vs 0.63, rate ratio 0.662; P = 0.047). Over 52 weeks, the exacerbation rate was reduced by 50% (P < 0.001). Omalizumab had an acceptable safety profile, with no statistically significant (P < 0.05) differences in adverse events observed between omalizumab and placebo. CONCLUSION: Add-on omalizumab is well-tolerated and reduces exacerbations in children (6-<12 years) with severe persistent allergic asthma, inadequately controlled despite high-dose ICS plus a LABA. It should be noted that the sample size was not based on providing statistical power in the severe subgroup, and no corrections were made for multiple comparisons; however, outcomes consistently favoured omalizumab.

Unexpected cross-reactivity in a cat-allergy patient. An allergic reaction at the circus

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