Alterations in TP53, cyclin D2, c-Myc, p21WAF1/CIP1 and p27KIP1 expression associated with progression in B-CLL.

B-cell chronic lymphocytic leukaemia (B-CLL) originates from B lymphocytes that may differ in the activation level, maturation state or cellular subgroups in peripheral blood. Tumour progression in CLL B cells seems to result in gradual accumulation of the clone of resting B lymphocytes in the early phases (G0/G1) of the cell cycle. The G1 phase is impaired in B-CLL. We investigated the gene expression of five key cell cycle regulators: TP 53, c-Myc, cyclin D2, p21WAF1/CIP1 and p27KIP1, which primarily regulate the G1 phase of the cell cycle, or S-phase entry and ultimately control the proliferation and cell growth as well as their role in B-CLL progression. The study was conducted in peripheral blood CLL lymphocytes of 40 previously untreated patients. Statistical analysis of correlations of TP53, cyclin D2, c-Myc, p21WAF1/CIP1 and p27KIP1 expressions in B-CLL patients with different Rai stages demonstrated that the progression of disease was accompanied by increases in p53, cyclin D2 and c-Myc mRNA expression. The expression of p27KIP1 was nearly statistically significant whereas that of p21 WAF1/CIP1 showed no such correlation. Moreover, high expression levels of TP53 and c-Myc genes were found to be closely associated with more aggressive forms of the disease requiring earlier therapy

Ekspresja podoplaniny w raku jasnokomórkowym jajnika

Abstract Introduction: Podoplanin is a transmembrane glycoprotein expressed in endothelial lymphatic cells. It was proven to be a predictive marker in a variety of cancers e.g. mesothelioma and head and neck squamous-cell carcinoma. Ovarian clear cell carcinoma (OCCC) is a rare and unique histopathologic subtype of epithelial ovarian cancer (EOC). The molecular basis of that phenomenon remains unknown. Objectives: The aim of our study was to assess podoplanin expression on the protein level in OCCC. Material and Methods: Immunohistochemistry was performed on paraffin-embedded tissues from 19 patients with diagnosed OCCC. Results: Podoplanin expression was present (moderate or strong) in 52% of OCCC cases (10/19). Nine of eleven (81,2%) postmenopausal and one of eight (12,5%) premenopausal women were podoplanin positive. No differences in podoplanin expression were found in relation to clinical features of the tumor. Conclusion: The incidence of podoplanin expression is higher in ovarian clear cell adenocarcinoma in postmenopausal patients.Streszczenie Wstęp: Podoplanina jest przezbłonową glikoproteiną występującą w komórkach śródbłonka naczyń limfatycznych. Swoje zastosowanie jako marker predykcyjny znalazła w diagnostyce międzybłoniaka czy też w raku głowy i szyi. Jasnokomórkowy rak jajnika (OCCC) jest rzadko występującym i odmiennym histopatologicznym podtypem nabłonkowego raka jajnika. Molekularne podłoże tego zjawiska nadal nie jest znane. Cel: Celem badań była ocena ekspresji podoplaniny na poziomie białka w jasnokomórkowym raku jajnika. Materiał i metody: Ekspresję podoplaniny oceniono metodą immunohistochemiczną z zastosowaniem techniki macierzy tkankowych (TMA) u 19 pacjentek z OCCC. Wyniki: Ekspresja podoplaniny była obecna (średnia lub wysoka) w 52% przypadków OCCC (10/19). Jej ekspresję wykazano u jednej Spośród ośmiu pacjentek przed menopauzą (12,5%) i u 9 spośród 11 (81,2%) po menopauzie. Nie wykryto różnic w ekspresji podoplaniny w odniesieniu do cech klinicznych nowotworu. Wnioski: Częsta ekspresja podoplaniny jest charakterystyczna dla kobiet po menopauzie ze zdiagnozowanym rakiem jasnokomórkowym jajnika

Impaired endotoxin tolerance on example of IL6 in B-CLL

IL-6 uznaje się za kluczową cytokinę zaangażowaną w mechanizmy obronne organizmu, w procesach krwiotworzenia i reakcjach zapalnych, o wielokierunkowym działaniu auto- i parakrynnym. Jej produkcja jest zazwyczaj przemijająca i ściśle regulowana. Stosując metodę RT-PCR w czasie rzeczywistym (real time PCR) zbadano ekspresję IL-6 na poziomie mRNA, w prawidłowych i białaczkowych limfocytach B spoczynkowych i stymulowanych LPS przez 30' i 24 h. Poziom ekspresji IL-6 wyrażony wartością RQ w puli limfocytów PBL-B, w porównaniu z subpopulacją limfocytów B zdrowych dawców, był istotnie statystycznie niższy, wartość p=0,00348. Po 24 h w prawidłowych limfocytach B poziom RQ mRNA IL-6 istotnie statystycznie obniżał się (p=0,012970), natomiast w subpopulacji PBL-B istotnie statystycznie nadal wzrastał (p=0,000001). Uzyskane dane wydają się wskazywać na zaburzanie tolerancji endotoksycznej w PBL-BIL-6 is considered a key cytokine involved in immune organism response, hematopoietic processes and inflammatory reactions, based on multidirectional auto and paracrine auction Its production is usually transient and tightly regulated.By the means of Real Time PCR we studied IL-6 expression on mRNA level (RQ) in B-CLL lymphocytes and in CD19+ subpopulation of normal B lymphocytes before and after 30 min and 24h LPS stimulation. The IL-6 expression level (RQ) in B-CLL cells was significantly lower (p=0.00348) compared with the subpopulation of CD19+ normal lymphocytes. 30 min LPS stimulation resulted in increase of IL6 mRNA expression, both in normal and B-CLL lymphocytes. IL6 mRNA expression level decreased after 24h LPS stimulation (p=0.012970) in normal lymphocytes, whereas B-CLL cells showed constant increase (p=0.000001). These data seem to indicate an aberrant endotoxin tolerance in B-CLL

Different expression of CD180, CD284 and CD14 receptors on the CD19+ subpopulation of normal and B-CLL lymphocytes.

Numerous experimental data indicate that B-CLL development and progression are influenced by antigenic pressure. It can not be excluded that these antigens may originate from bacteria and viruses. Toll like receptors (TLRs) interact with pathogen associated molecular patterns as part of innate immunity. TLRs are currently used to target different subclasses of B-cell leukemia, and TLR agonists are being evaluated in clinical trials. It is little known regarding the repertoire and function of TLR in B-CLL. The aim of the study was to assess the CD180, CD284 and mCD14 levels in CD19+ subpopulation of B-CLL peripheral blood lymphocytes and compare them with respective levels in the normal B-cells of adult volunteers, before and after LPS stimulation. We investigated the percentage of the CD19+CD180+, CD19+CD284+, CD19+CD14+ cells and the mean fluorescence intensity (MFI) of CD180, CD284 and CD14 antigens among CD19+ B-CLL as well as in the normal B cells for comparison. MFI analysis revealed that CD180, CD284 and CD14 expression was higher on normal B cells then on CD19+ B-CLL (MFI CD180: 99.16 vs. 25.3, MFI CD284: 7.37 vs. 5.79 and MFI CD14 25.07 vs. 8.32). After 24-hour LPS activation of B-cells, CD180 MFI appeared to decrease, in both healthy and B-CLL patients. CD284 MFI in healthy controls decreased after LPS stimulation while slight increase of MFI was observed in leukemic cells. CD14 MFI in leukemic cells was moderately higher after LPS in comparison to CD14 MFI without LPS stimulation, whereas CD14 MFI in normal CD19+ cells after LPS stimulation decreased over three times. Variations observed in expression of both normal and leukemic receptors may be due to their different sensitivity to antigenic stimulation

Ocena wybranych parametrów klinicznych i biochemicznych u kobiet chorych na raka jajnika w okresie 5-letniej obserwacji

Introduction: Ovarian cancer is a neoplasm characterised by considerable mortality. The aim of numerousstudies which have been conducted is to determine the impact of clinical and biochemical parameters onovarian cancer prognosis, survival and treatment. Serum CA 125 is mainly used to monitor treatment, detectrecurrence and predict response to chemotherapy. Increased epidermal growth factor receptor (EGFR) cancertissue expression facilitates cell proliferation, apoptosis inhibition and increase of angiogenesis. Material and methods: A group of 74 patients treated with surgery and chemotherapy due to ovarian cancerwere included in the study. The analysis of clinical parameters (age, tumour histology and grading, clinical stage according to FIGO, type of surgery) as well as CA 125 concentration and EGFR cancer tissue expression in relationto 5-year survival rates was performed. Results: 5-year survival rates were significantly related to: histological grading, clinical FIGO stage, theextent of primary surgery and cancer recurrence. The best predictors of increased survival rates in ovariancancer patients were: high histological grade, low clinical FIGO stage, optimal debulking and no recurrence.Statistical multivariate analysis did not demonstrate the influence of EGFR on 5-year survival rate in ovariancancer patients. However, the considerable and unequivocal impact of the last result of CA 125 concentration,the extent of surgical treatment, histological diagnosis and clinical FIGO stage, in relation to 5-year survival rate,was demonstrated. Conclusion: The results suggest a considerable and significant impact of CA 125 concentration, histologicaldiagnosis, clinical FIGO stage, and optimal debulking, on the 5-year survival rates of patients with ovarian cancer

Ekspresja genu podoplaniny w złośliwych nowotworach jajnika

Introduction: Ovarian cancer is mostly diagnosed in postmenopausal women. The hormonal microenvironmentof ovarian tumour development in pre- and postmenopausal women is different. The environment mayinfluence the molecular basis of ovarian cancer. Podoplanin is a proven marker of lymphangiogenesis and isengaged in tumour progression and metastasis. Objectives: The aim of our study was to assess podoplanin expression at the mRNA level in ovarian serousand mucinous adenocarcinoma. Material and methods: mRNA expression of podoplanin was assessed in 42 ovarian cancer patients bymeans of real-time PCR. Results: Podoplanin was expressed at the mRNA level in all ovarian cancer patients. It was significantlylower (p < 0.001) in cancer tissues (RQ = 4.46) than in healthy control (RQ = 13.03). No significant differenceswere found in podoplanin expression either between pre- and postmenopausal women or in relation to FIGOand grading. Conclusion: Decreased podoplanin expression is characteristic for serous and mucinous ovarianadenocarcinoma

Gastric lipomatosis

Gastric lipomatosis is a condition characterized by the presence of multiple lipomas or diffuse mature adipose tissue infiltration within the gastric wall. The diffuse form is thought to be an extremely rare, with only few described cases. The lesion may be asymptomatic or associated with symptoms and signs depending on location and size. Treatment depends on clinical presentation, range and complications. In a symptomatic disease, it should be surgical, but conservative treatment is preferred for asymptomatic and solitary lesions. Among diagnostic methods, computed tomography and magnetic resonance imaging are thought to be the most valuable

Transcriptome Sequencing of the Spleen Reveals Antiviral Response Genes in Chickens Infected with CAstV

Astrovirus infections pose a significant problem in the poultry industry, leading to multiple adverse effects such as a decreased egg production, breeding disorders, poor weight gain, and even increased mortality. The commonly observed chicken astrovirus (CAstV) was recently reported to be responsible for the “white chicks syndrome” associated with an increased embryo/chick mortality. CAstV-mediated pathogenesis in chickens occurs due to complex interactions between the infectious pathogen and the immune system. Many aspects of CAstV–chicken interactions remain unclear, and there is no information available regarding possible changes in gene expression in the chicken spleen in response to CAstV infection. We aim to investigate changes in gene expression triggered by CAstV infection. Ten 21-day-old SPF White Leghorn chickens were divided into two groups of five birds each. One group was inoculated with CAstV, and the other used as the negative control. At 4 days post infection, spleen samples were collected and immediately frozen at −70 °C for RNA isolation. We analyzed the isolated RNA, using RNA-seq to generate transcriptional profiles of the chickens’ spleens and identify differentially expressed genes (DEGs). The RNA-seq findings were verified by quantitative reverse-transcription PCR (qRT-PCR). A total of 31,959 genes was identified in response to CAstV infection. Eventually, 45 DEGs (p-value 2 fold change > 1) were recognized in the spleen after CAstV infection (26 upregulated DEGs and 19 downregulated DEGs). qRT-PCR performed on four genes (IFIT5, OASL, RASD1, and DDX60) confirmed the RNA-seq results. The most differentially expressed genes encode putative IFN-induced CAstV restriction factors. Most DEGs were associated with the RIG-I-like signaling pathway or more generally with an innate antiviral response (upregulated: BLEC3, CMPK2, IFIT5, OASL, DDX60, and IFI6; downregulated: SPIK5, SELENOP, HSPA2, TMEM158, RASD1, and YWHAB). The study provides a global analysis of host transcriptional changes that occur during CAstV infection in vivo and proves that, in the spleen, CAstV infection in chickens predominantly affects the cell cycle and immune signaling

Different expression of CD180, CD284 and CD14 receptors on the CD19+ subpopulation of normal and B-CLL lymphocytes.

Numerous experimental data indicate that B-CLL development and progression are influenced by antigenic pressure. It can not be excluded that these antigens may originate from bacteria and viruses. Toll like receptors (TLRs) interact with pathogen associated molecular patterns as part of innate immunity. TLRs are currently used to target different subclasses of B-cell leukemia, and TLR agonists are being evaluated in clinical trials. It is little known regarding the repertoire and function of TLR in B-CLL. The aim of the study was to assess the CD180, CD284 and mCD14 levels in CD19+ subpopulation of B-CLL peripheral blood lymphocytes and compare them with respective levels in the normal B-cells of adult volunteers, before and after LPS stimulation. We investigated the percentage of the CD19+CD180+, CD19+CD284+, CD19+CD14+ cells and the mean fluorescence intensity (MFI) of CD180, CD284 and CD14 antigens among CD19+ B-CLL as well as in the normal B cells for comparison. MFI analysis revealed that CD180, CD284 and CD14 expression was higher on normal B cells then on CD19+ B-CLL (MFI CD180: 99.16 vs. 25.3, MFI CD284: 7.37 vs. 5.79 and MFI CD14 25.07 vs. 8.32). After 24-hour LPS activation of B-cells, CD180 MFI appeared to decrease, in both healthy and B-CLL patients. CD284 MFI in healthy controls decreased after LPS stimulation while slight increase of MFI was observed in leukemic cells. CD14 MFI in leukemic cells was moderately higher after LPS in comparison to CD14 MFI without LPS stimulation, whereas CD14 MFI in normal CD19+ cells after LPS stimulation decreased over three times. Variations observed in expression of both normal and leukemic receptors may be due to their different sensitivity to antigenic stimulation