The Hippo pathway in colorectal cancer

Colorectal cancer (CRC) is one the most frequently diagnosed neoplastic diseases worldwide. Currently, aside from traditional chemotherapy, advanced CRCs are treated with modern drugs targeting cellular components such as epithelial growth factor receptor (EGFR). Since up to 70% of metastasized CRCs are drug resistant, the description of recent progress in cellular homeostasis regulation may shed new light on the development of new molecular targets in cancer treatment. The Hippo pathway has recently become subject of intense investigations since it plays a crucial role in cell proliferation, differentiation, apoptosis and tumourigenesis. Components of the Hippo pathway are deregulated in various human malignancies, and expression levels of its major signal transducers were proposed as prognostic factors in colorectal cancer. In this review we focused on recent data regarding Hippo pathway, its up-stream signals and down-stream effectors. Hippo negatively regulates its major effectors, YAP1 and TAZ kinases, which act as transcriptional co-activators inducing expression of genes involved not only in tissue repair and proliferation but are also oncoproteins involved in tumour development and progression. The deregulation of Hippo pathway components was found in many malignancies. The interactions between Hippo and Wnt/β-catenin signalling, crucial in the maintenance of cell homeostasis, have been described in relation to the control of intestinal stem cell proliferation and CRC development. The recently discovered positive feedback loop between activated YAP1 and increased EGFR/KRAS signalling found in oesophageal, ovarian and hepatocellular cancer has been related to the CRC progression and resistance to EGFR inhibitors during CRC therapy

APPLICATION OF MULTI-CRITERIA ANALYSIS BASED ON THE INDIVIDUAL PSYCHOLOGICAL PROFILE FOR RECOMMENDER SYSTEMS

This paper presents a novel approach for user classification exploiting multicriteriaanalysis. This method is based on measuring the distance between anobservation and its respective Pareto front. The obtained results show that thecombination of the standard KNN classification and the distance from Paretofronts gives satisfactory classification accuracy – higher than the accuracy obtainedfor each of these methods applied separately. Conclusions from thisstudy may be applied in recommender systems where the proposed methodcan be implemented as the part of the collaborative filtering algorithm

Cell-penetrating peptides as a promising tool for delivery of various molecules into the cells

Many biologically active compounds, including macromolecules that are used as various kinds of drugs, must be delivered to the interior of cell or organelles such as mitochondria or nuclei to achieve a therapeutic effect. However, very often, lipophilic cell membrane is impermeable for these molecules. A new method in the transport of macromolecules through the cell membrane is the one based on utilizing cell-penetrating peptides (CPPs). Invented 25 years ago, CPPs are currently the subject of intensive research in many laboratories all over the world. CPPs are short compounds comprising up to 30 amino acid residues, which penetrate the cell membrane but do not cause cell damage. Additionally, CPPs can transfer hydrophilic molecules (peptides, proteins, nucleic acids) which exceed their mass, and for which the cell membrane is generally impermeable. In this review, we concentrate on the cellular uptake mechanism of CPPs and a method of conjunction of CPPs to the transported molecules. We also highlight the potential of CPPs in delivering various kinds of macromolecules into cells, including compounds of therapeutic interest

The expression of particular glucose transporters and insulin resistance indicators in the risk groups of type 2 diabetes — a two-year follow-up

Introduction: The proper expression of particular glucotransporter (GLUT) isoforms determines a sufficient supply of glucose to tissues. The impairment of cellular glucose transport observed in insulin resistance leads to glucose metabolism disturbances. The aim of this study was the estimation of insulin resistance indicators and the quantitative expression of GLUT-1, GLUT-3 and GLUT-4 on peripheral blood lymphocytes in prediabetic subjects and persons with a positive family history of type 2 diabetes during 24 months of observation. Material and methods: The study included 25 prediabetic subjects (according to WHO criteria) and 24 normoglycaemic individuals with a positive family history of type 2 diabetes. Twenty three healthy subjects with no family history of type 2 diabetes, matched with BMI, served as a control group. All participants were recommended to perform physical activity for at least 140 minutes per week and to maintain a low calorie diet. The peripheral blood lymphocytes demonstrating expression of GLUT-1, GLUT-3 and GLUT-4 were labelled with the use of indirect immunofluorescence. The expression of GLUT isoforms was investigated by flow cytometry. Cells were stained by using anti-human GLUT antibodies and FITC-conjugated immunoglobulin. Flow cytometry was performed using a FACS Calibur (Becton-Dickinson). Additionally, we determined: fasting plasma glucose (FPG), insulin and C peptide concentrations, HOMA-IR, BMI and WHR. All the tests were performed at baseline, and after 12 and 24 months. Results: At baseline, prediabetics and subjects with a positive family history of type 2 diabetes were characterised by a much higher expression of GLUT-4 compared to control subjects. Twenty four months of lifestyle modification resulted in significant lowering of the expression of GLUT-4 on the surface of PBL in both studied groups, with no differences in the expression of GLUT-1 or GLUT-3. Both prediabetic subjects and individuals with a positive family history of type 2 diabetes revealed no significant differences in determined insulin resistance markers after 24 months of the observation compared to the baseline values. Conclusions: The estimation of typical GLUT isoforms present on the peripheral blood lymphocytes, as well as the evaluation of insulin resistance indicators, are obviously insufficient for monitoring the metabolic disorders progression in the risk groups of type 2 diabetes. The decrease in GLUT-4 lymphocyte expression may reflect a positive influence of lifestyle modification on a tissue redistribution of this crucial insulin-dependent glucotransporter. The determination of GLUT-4 on the surface of peripheral blood lymphocytes can be a useful tool for the evaluation of the efficacy of therapeutic actions in subjects at high risk of type 2 diabetesWstęp: Właściwa ekspresja poszczególnych izoform glukotransporterów (GLUT) warunkuje odpowiednie zaopatrzenie tkanek w glukozę. Upośledzenie dokomórkowego transportu glukozy obserwowane w warunkach insulinooporności prowadzi do zaburzeń metabolizmu glukozy. Celem badania była ocena wskaźników insulinooporności oraz ilościowej ekspresji GLUT-1, GLUT-3 i GLUT-4 na limfocytach krwi obwodowej osób ze stanem przedcukrzycowym oraz osób z dodatnim wywiadem rodzinnym w kierunku cukrzycy typu 2 w trakcie 24-miesięcznej obserwacji. Materiał i metody: Do badania włączono 25 osób ze stanem przedcukrzycowym (wg kryteriów WHO) i 24 osoby z normoglikemią z dodatnim wywiadem rodzinnym w kierunku cukrzycy typu 2. Grupę kontrolną stanowiły 23 zdrowe osoby bez wywiadu rodzinnego w kierunku cukrzycy, dobrane pod względem BMI. Wszystkim uczestnikom badania zalecono zwiększenie aktywności fizycznej do co najmniej 140 min tygodniowo oraz stosowanie diety niskokalorycznej. Do znakowania limfocytów krwi obwodowej wykazujących ekspresję GLUT-1, GLUT-3 i GLUT-4 użyto techniki immunofluorescencji pośredniej z wykorzystaniem swoistych przeciwciał anty-ludzkich GLUT oraz immunoglobulin sprzężonych z FITC. Ekspresję poszczególnych izoform GLUT oznaczano za pomocą cytometrii przepływowej przy użyciu cytometru FACS Calibur (Becton-Dickinson) Dodatkowo oznaczano glikemię na czczo, stężenie insuliny i C-peptydu na czczo, wskaźniki HOMA-IR, BMI oraz WHR. Wszystkie oznaczenia były wykonywane wyjściowo, po 12 i po 24 miesiącach obserwacji. Wyniki: W chwili rozpoczęcia badania osoby ze stanem przedcukrzycowym oraz osoby z dodatnim wywiadem rodzinnym w kierunku cukrzycy typu 2 charakteryzowały się znacznie wyższą ekspresją GLUT-4 w porównaniu z grupą kontrolną. W czasie 24 miesięcy od wdrożenia modyfikacji stylu życia zaobserwowano istotny spadek ekspresji GLUT-4 na limfocytach krwi obwodowej w obu badanych grupach ryzyka, przy braku różnic w ekspresji GLUT-1 i GLUT-3. Zarówno w grupie osób ze stanem przedcukrzycowym, jak i wśród osób z dodatnim wywiadem rodzinnym w kierunku cukrzycy typu 2 po 24 miesiącach nie stwierdzono istotnych statystycznie różnic w zakresie badanych wskaźników insulinopoorności w porównaniu z wartościami wyjściowymi. Wnioski: Ani oznaczanie typowych dla limfocytów krwi obwodowej izoform GLUT, ani ocena wskaźników insulinooporności są niewystarczające do monitorowania progresji zaburzeń metabolicznych w grupach ryzyka cukrzycy typu 2. Spadek ekspresji GLUT-4 na limfocytach jest prawdopodobnie odzwierciedleniem pozytywnego wpływu modyfikacji stylu życia na tkankową redystrybucję tego kluczowego insulinozależnego transportera glukozy. Oznaczanie GLUT-4 na limfocytach krwi obwodowej może być wartościowym narzędziem do oceny skuteczności interwencji terapeutycznych w grupach ryzyka cukrzycy typu 2

Characteristics of dental pulp in human upper first premolar teeth based on immunohistochemical and morphometric examinations

Teeth extracted for orthodontic reasons are commonly considered as healthy. Therefore, it is possible to examine structure of the dental pulp can be fully recognized and how it is affected by malocclusion. The aim of the study was to evaluate by immunohistochemistry (IHC) and morphometry dental pulp in human upper first premolar teeth extracted for orthodontic reasons. The material comprised 36 teeth of 20 patients in the age range 16–26 years. By the use of IHC markers the presence of immunocompetent cells (CD20, CD45RO, and CD68), blood vessels (CD31) and nerves (PGP9.5) were examined in the pulp. Inflammatory infiltrates and tissue atrophy were observed in 24 and 10 teeth, respectively. Strong positive correlation between the width of the odontoblastic layer, the number of rows of odontoblast nuclei and the increase of MVA (microvessel area) in the pulp of atrophic teeth was found. The cellular infiltrations found in H&E-stained sections were identified by IHC as memory T cells (CD45RO+) and B lymphocytes (CD20+) with macrophages (CD68+) present at the periphery. The CD20 antigen was intensively expressed in 13 teeth, CD45RO in 33 teeth, and CD68 in 20 teeth. Thus, despite the lack of any clinical signs of pulp disease many teeth extracted for orthodontic reasons show focal pulp inflammation and atrophy which probably results from the malocclusion stress accompanying teeth crowding

Interrupted orthodontic force results in less root resorption than continuous force in human premolars as measured by microcomputed tomography

Introduction. Root resorption is an undesirable but very frequently occurring sequel of orthodontic treatment. The aim of this study was to compare root resorption caused by either continuous (CF) or interrupted (IF) orthodontic force. Material and methods. The study was performed on human subjects on 30 first upper and lower premolars scheduled for extraction for orthodontic reasons. During four weeks before extraction 12 teeth were subjected to either CF or IF. The force was generated by a segmental titanium-molybdenum alloy cantilever spring that was activated in buccal direction. Initially a force of 60 CentiNewton was used in both CF and IF groups, the force in the former, however, was reactivated every week for 4 weeks. There was no reactivation of force in the IF group after initial application. A morphometric analysis of root resorption was performed by microcomputed tomography and the extent of tooth movement was measured on stone casts. Furthermore, a Tartarate-Resistant Acidic Phosphatase activity (TRAP), the marker enzyme of osteoclasts and cementoclasts, was determined by histochemical method. The Mann-Whitney U test was used to compare the difference in measured parameters between treatment and control tooth groups. Results. The number of resorption craters was significantly higher and their average volume almost twice as large in the CF compared to the IF group (p < 0.05). However, the distance of tooth displacement was similar for both groups. Cementoclasts were detected with the TRAP technique on the surface of two teeth only; both were subjected to continuous force. Conclusions. The use of IF leads to less destruction of root structure as opposed to continuous force while the same tooth movement was achieved

The influence of sense of coherence on emotional response in heart transplant recipients : a preliminary report

INTRODUCTION: The success of heart transplantation in prolonging life and well-being must be considered in reference to its psycho-social outcomes, which intrinsically affect the long-term post-transplant morbidity. Sense of coherence and emotional response to organ reception are important factors in this group of patients. THE AIM OF THIS STUDY: The aim of this study was to assess the contribution of sense of coherence to emotional response to transplantation in heart transplant recipients. MATERIAL AND METHODS: The study was conducted on a group of 46 heart transplant recipients. The following research tools were applied in the assessment of personal resources (sense of coherence) and emotional response to heart transplant surgery: the Sense of Coherence Questionnaire developed by Antonovsky (SOC-29) and the Transplant Effects Questionnaire (TxEQ). The data were analyzed statistically. RESULTS: Heart transplant recipients do not experience guilt toward the donors and have no difficulties in disclosing their identities as heart transplant recipients. The study reports good adherence to immunosuppressive treatment recommendations and both a moderate concern about and a sense of responsibility for the transplanted organs among the patients. Global SOC was associated with guilt toward the donor, concern about the transplanted heart, and disclosure of the recipient's identity. CONCLUSIONS: The strength of the patients’ global sense of coherence is related to the level of their emotional response to the heart transplant surgery

Cellular glucose transport disturbances as a marker of the pre-diabetic state - pathogenetic and clinical significance of the assessment of GLUT4 expression

Wstęp: Glukotransporter 4 (GLUT4) jest przedstawicielem rodziny integralnych białek błonowych transportujących glukozę do komórek na drodze dyfuzji ułatwionej. W dostępnym piśmiennictwie stwierdzano zwiększoną ekspresję białka GLUT4 na powierzchni leukocytów osób chorych na cukrzycę; nie ma natomiast publikacji dotyczących badania ekspresji glukotransporterów u osób ze stanem przedcukrzycowym. Celem pracy było porównanie ilościowej ekspresji GLUT4 na limfocytach krwi obwodowej u osób należących do grup ryzyka cukrzycy typu 2 oraz u osób zdrowych. Materiał i metody: Do badania włączono 15 osób ze stanem przedcukrzycowym oraz 15 osób metabolicznie zdrowych z dodatnim wywiadem rodzinnym w kierunku cukrzycy typu 2 (pokrewieństwo pierwszego stopnia). Grupę kontrolną stanowiło 15 osób zdrowych z negatywnym wywiadem rodzinnym. Do znakowania limfocytów wykazujących ekspresję białka GLUT4 użyto techniki immunofluorescencji pośredniej. Ilościowego oznaczenia GLUT4 na komórkach dokonano przy użyciu cytometru przepływowego. Wyniki: Ekspresja GLUT4 w grupie kontrolnej wynosiła 12 ± 1,5% i była znamiennie niższa od ekspresji GLUT4 w grupie osób ze stanem przedcukrzycowym (18,2 ± 8,8%, p = 0,008) oraz z dodatnim wywiadem rodzinnym (17,9 ± 9%, p = 0,01). Wnioski: Zwiększona ekspresja GLUT4 u osób z dodatnim wywiadem rodzinnym w kierunku cukrzycy typu 2 sugeruje istnienie zaburzeń dokomórkowego transportu glukozy w fazie przedhiperglikemicznej. Wydaje się, że oznaczenie ekspresji GLUT4 u osób z grup zwiększonego ryzyka może być wartościowym testem do wczesnego wykrywania cukrzycy. (Endokrynol Pol 2010; 61 (3): 269-274)Introduction: GLUT4 is a representative of the family of integral membrane proteins which facilitate glucose transport across cellular membranes. In the available literature there is no publication referring to the investigations of glucotransporter expression in pre-diabetic subjects. However, GLUT4 protein overexpression was shown in leukocytes of diabetic patients. The aim of this study was to compare GLUT4 quantitative expression in peripheral blood lymphocytes in type 2 diabetes mellitus risk groups to healthy subjects. Material and methods: The study groups included 15 pre-diabetic subjects and 15 persons with normal glucose tolerance and positive family history of type 2 diabetes mellitus (first-degree relatives). As a control group, 15 healthy persons with no family history of diabetes were enrolled. The expression of GLUT4 on the surface of peripheral blood lymphocytes was investigated with the use of indirect immunofluorescence. Quantitative determination of GLUT4 was performed with the use of flow cytometry. Results: In the control group, GLUT4 expression amounted to 12 ± 1.5% and was significantly lower in relation to both pre-diabetic subjects (18.2 ± 8.8%; p = 0.008) and the positive family history group (17.9 ± 9%; p = 0.001). Conclusions: GLUT4 overexpression in subjects with positive family history of type 2 diabetes mellitus suggests that cellular glucose transport disturbances occur prior to hyperglycaemia. Determination of GLUT4 expression appears to be a possibly useful method of early detection in individuals at high risk of diabetes. (Pol J Endocrinol 2010; 61 (3): 269-274

Decreased expression of p73 in colorectal cancer

Introduction. The colorectal cancer (CRC) is one of the most frequent cancer in Poland and worldwide. This disease is characterized by distinct genetic alterations. p73 belongs to the p53 gene family; however, its role in the pathogenesis of CRC has not been completely understood. p73 gene encodes several mRNA variants and protein isoforms with its longest and fully functional p73a (mRNA) and TAp73a (protein) isoform. The aim of the study was to investigate p73 gene expression at the mRNA (p73a) and protein (TAp73a) levels in CRC. Material and methods. Small sections of the crc tumor tissue and macroscopically unchanged colon mucosa and submucosa from the dissection margin were collected from 23 patients diagnosed with CRC. p73 mRNA levels were measured by Real-time PCR (QPCR) method and the expression level of TAp73a protein was assessed by Western blotting (WB) and immunohistochemical (IHC) staining. Results. We found a 37% decrease in the level of p73a mRNA in neoplastically changed (tumor) compared with unchanged normal colon tissue from the surgical margin (p = 0.041). No correlations were found between mRNA levels in cancer tissue and clinical-pathological parameters. The semi-quantification of TAp73a protein revealed lower and higher TAp73a protein contents in 11/23 and 12/23 of tumor samples, respectively, when compared with the median value of TAp73a protein in normal colon tissue (p = 0.61). The level of TAp73a protein level was 5 times lower in poorly differentiated cancer cells (G3) in comparison to moderately differentiated ones (G2; p = 0.02). No statistically significant correlations were observed between the level of the TAp73a protein and clinical-pathological patients’ characteristics. The IHC analysis of TAp73a protein presence in CRC samples showed decreased immunoreactivity when compared with matched sections of the unchanged colon wall in 4/9 patients, similar intensity of the IHC reaction in 4/9 patients and increased immunoreactivity in 1/9 patients. The TAp73a protein was localized mainly in the cytoplasm of the cancer cells. No statistically significant correlations between IHC results and clinical-pathological features of the patients were found. Conclusions. The obtained results suggest that the p73 gene may play a role as a tumor suppressor in the CRC progression