Ultrasound differentiation between benign versus malignant adnexal masses in pregnant patients.

OBJECTIVES: The aim of this study was to assess the performance of the International Ovarian Tumor Analysis (IOTA) group ultrasound Simple Rules method in differentiating between benign and malignant ovarian tumors in pregnant patients. MATERIAL AND METHODS: A prospective observational study that involved pregnant patients referred to our center due to suspicions of ovarian masses between January 2015 and December 2017 was performed. The Simple Rules performance was evaluated against the histopathological results. Each of the 10 sonographic Simple Rules were computed by logistic regression to demonstrate their odds ratios in predicting malignancy. RESULTS: Ultrasound were conducted in 153 subjects, and 61 of those patients underwent surgery. By assigning masses presenting inconclusive picture as probably malignant, the Simple Rules method showed sensitivity of 91.67% and specificity of 69.39%. After exclusion of masses with inconclusive findings, the method showed sensitivity of 87.5% and specificity of 94.44%. The Simple Rules risk estimation method for the 1% risk cutoff showed sensitivity of 100% and specificity of 51.02%. For the 3% cutoff, sensitivity was 91.67% and specificity was 53.06%. And for 30 % cutoff, sensitivity was 91.67% and specificity 73.47 %. The logistic regression model showed that the M-rules increased the risk of malignancy while the B-rules decreased the risk. CONCLUSIONS: Most ovarian masses in pregnant patients may be correctly categorized as benign or malignant using Simple Rules. This protocol may facilitate the management of pregnant patients presenting with adnexal masses.

A Prospective Pilot Study on Use of Liquid Crystal Thermography to Detect Early Breast Cancer.

Background: Breast cancer is the most common cancer in women. While mammography is the standard for early detection in women older than 50 years of age, there is no standard for younger women. The aim of this prospective pilot study was to assess liquid crystal contact thermography, using the Braster device, as a means for the early detection of breast cancer. The device is intended to be used as a complementary tool to standard of care (sonography, mammography, etc). Patients and Methods: A total of 274 consecutive women presenting at Polish breast centers for prophylactic breast examination were enrolled to receive thermography; 19 were excluded for errors in thermographic image acquisition. The women were divided according to age (n = 135, <50 years; n = 120, ≥50 years). A control population was included (n = 40, <50 years; n = 23, ≥50 years). The primary endpoint, stratified by age group, was the C-statistic for discrimination between breast cancer and noncancer. Results: In women with abnormal breast ultrasound (n = 95, <50 years; n = 87, ≥50 years), the C-statistic was 0.85 and 0.75, respectively (P = .20), for discrimination between breast cancer and noncancer. Sensitivity did not differ (P = .79) between the younger (82%) and older women (78%), while specificity was lower in the older women (60% vs 87%, P = .025). The false-positive rate was similar in women with normal and abnormal breast ultrasound. Positive thermographic result in women with Breast Imaging Reporting and Data System (BIRADS) 4A on ultrasound increased the probability of breast cancer by over 2-fold. Conversely, a negative thermographic result decreased the probability of cancer more than 3-fold. Breast size and structure did not affect the thermography performance. No adverse events were observed. Conclusions: Thermography performed well in women <50 years of age, while its specificity in women ≥50 years was inadequate. These promising findings suggest that the Braster device deserves further investigation as a supporting tool for the early detection of breast cancer in women younger than 50 years of age.

Imatinib inhibits the regrowth of human colon cancer cells after treatment with 5-FU and cooperates with vitamin D analogue PRI-2191 in the downregulation of expression of stemness-related genes in 5-FU refractory cells.

Conventional cytotoxic drugs preferentially eliminate differentiated cancer cells but spare relatively more resistant stem-like cancer cells capable to initiate recurrence. Due to cancer cell plasticity, the stem-like phenotype can be also acquired by cancer cells refractory to treatment with cytotoxic drugs. We investigated whether drugs inhibiting receptor tyrosine kinases could be used to target human colon cancer cells initiating cancer regrowth following conventional cytotoxic treatment. The moderately differentiated cell line HT-29 and poorly differentiated cell line HCT-116 were exposed to 5-fluorouracil (5-FU). Cells that resisted the exposure to 5-FU were subsequently treated with imatinib or sunitinib. Both drugs reduced clonogenicity of 5-FU-refractory cells under normoxic and hypoxic culture conditions. The expression of numerous stemness-related genes was upregulated in cancer cells following the exposure to 5-FU, and remained at a high level in 5-FU-refractory cells undergoing renewal under normoxia, but decreased spontaneously under hypoxia. Imatinib downregulated the expression of stemness-related genes in cells undergoing renewal under normoxia. A combination of imatinib with PRI-2191, an analogue of 1,25-dihydroxyvitamin D3, downregulated stemness-related genes in HCT-116/5-FU cells more efficiently than imatinib alone. A synthetic analogue of 1,25-dihydroxyvitamin D2 (PRI-1906) abolished the effect of imatinib on gene expression in HCT-116/5-FU cells undergoing renewal under normoxia. Sunitinib promoted shift of phenotype of HT-29/5-FU cells undergoing renewal toward stem-like one. It suggests that the phenotype shift toward stemness induced by sequential sunitinib treatment following 5-FU treatment could increase a risk of cancer recurrence. In contrast to sunitinib, imatinib could be used both to interfere with cancer regrowth after conventional chemotherapy and to downregulate the expression of stemness-related genes in residual colon cancer cells capable to initiate cancer recurrence. The findings suggest that imatinib could also be combined with vitamin D analogue PRI-2191 to prevent recurrence more efficiently than imatinib alone and to compensate for vitamin D deficiency resulting from imatinib treatment.

Imatinib Inhibits the Renewal and Tumorigenicity of CT-26 Colon Cancer Cells after Cytoreductive Treatment with Doxorubicin.

Conventional anti-cancer drugs preferentially eliminate differentiated cancer cells but those cells that are spared (i.e. cancer stem cells: CSC), initiate recurrence. We tested whether drugs that target receptor tyrosine kinases (RTKs) involved in developmental signaling cascades and activated in CSC, could be used to silence and/or to eliminate colorectal cancer cells refractory to conventional treatment with cytoreductive drugs. A sequential treatment model was thereby developed with doxorubicin (DOX) and imatinib. CT-26 mouse colon carcinoma cells were pre-treated with DOX to select DOX-refractory cells with CSC properties, which were then subsequently treated with RTK inhibitor imatinib, where their regrowth was found to be inhibited. Under both normoxic and hypoxic conditions, imatinib potently inhibited clonogenicity of DOX-refractory CT-26 cells. Treatment with DOX did not eliminate tumorigenic CT-26 cells, since CT-26 cells pre-exposed to DOX in vitro, when inoculated subcutaneously, induced tumors in 90 % of mice, as opposed to a 100 % rate in the case of chemonaive CT-26 cells. In mice inoculated with chemonaive CT-26 cells, tumor formation was not prevented by imatinib. However, imatinib prevented tumor formation in 50 % of mice inoculated with CT-26 cells pre-exposed to DOX in vitro, with the remaining 50 % mice showing delayed tumor formation. These results suggest that the sequential use of the drug imatinib, as a drug targeting cancer cells expressing stem cell features after conventional cytoreductive treatment, is a promising future strategy for preventing tumor recurrence.

The Effect of Analogues of 1α,25-Dihydroxyvitamin D2 on the Regrowth and Gene Expression of Human Colon Cancer Cells Refractory to 5-Fluorouracil.

This study aimed to evaluate the capacity of hypocalcemic analogues of 1α,25-dihydroxyvitamin D2 (1,25D2) and 1α,25-dihydroxyvitamin D3 (1,25D3) to inhibit regrowth and regulate the stemness-related gene expression in colon cancer cells undergoing renewal after exposure to 5-fluorouracil (5-FU). All of the tested analogues of 1,25D2 equally potently decreased the clonogenicity and the proliferative activity of HT-29 cells which survived the exposure to 5-FU, but differently regulated gene expression of these cells during their renewal. 1,25D2 and analogues (PRI-1907 and PRI-1917), as well as 1,25D3 and analogue PRI-2191, decreased the relative expression level of several stemness-related genes, such as NANOG, OCT3/4, PROM1, SOX2, ALDHA1, CXCR4, in HT-29/5-FU cells during their renewal, in comparison to untreated HT-29/5-FU cells. The other 1,25D2 analogues (PRI-1906 and PRI-1916) were not capable of downregulating the expression of these stemness-related genes as the analogues PRI-1907 and PRI-1917 did. All of the tested vitamin D analogues upregulated CDH1, the gene encoding E-cadherin associated with epithelial phenotype. Out of the series of analogues studied, side-chain branched analogues of 1,25D2 (PRI-1907, PRI-1917) and the analogue of 1,25D3 (PRI-2191) might be used to target cancer cells with stem-like phenotypes that survive conventional chemotherapy.

Differential interference of vitamin D analogs PRI-1906, PRI-2191, and PRI-2205 with the renewal of human colon cancer cells refractory to treatment with 5-fluorouracil.

This study was aimed to determine whether hypocalcemic analogs of active forms of vitamins D modulate expression of genes related to stem-like phenotype in colon cancer cell lines HT-29 and HCT-116 undergoing renewal after the treatment with 5-fluorouracil (5-FU). Both lines express vitamin D receptor, but differ in differentiation stage and vitamin D sensitivity. Cells that resisted the 5-FU exposure were treated with synthetic analog of 1,25-dihydroxyvitamin D2 (PRI-1906) and analogs of 1,25-dihydroxyvitamin D3 (PRI-2191 and PRI-2205). Proliferative activity was more profoundly affected by vitamin D analogs in HT-29/5-FU than in HCT-116/5-FU cells. In HT-29/5-FU cells, analogs PRI-1906 and PRI-2191 downregulated the expression of genes related to survival, re-growth, and invasiveness during renewal, while PRI-2205 increased expression of genes related to differentiation only. In HCT-116/5-FU cells, PRI-2191 decreased the expression of stemness- and angiogenesis-related genes, whereas PRI-1906 augmented their expression. The effects in HCT-116/5-FU cells were observed at higher concentrations of the analogs than those used for HT-29/5-FU cells. Out of the series of analogs studied, PRI-2191 might be used to counteract the renewal of both moderately and poorly differentiated cancer cells following conventional treatment.

[Fertility-sparing surgical management of ovarian cancer]. / Leczenie chirurgiczne raka jajnika z zachowaniem plodnosci.

INTRODUCTION: Ovarian cancer (OC) continues to be one of the greatest challenges of modern oncology gynecology Most patients are diagnosed in the advanced stage of the disease, which requires aggressive and extensive surgical intervention. In approximately 8% of the cases, OC affects women <40 years of age. Among them, early-stage OC FIGO IA accounts for 40% of the affected individuals, which allows for fertility-sparing surgical management. OBJECTIVES: The aim of the study was to evaluate the efficacy of fertility-sparing management in OC women, with tumor limited to one ovary who wish to preserve their fertility MATERIAL AND METHODS: A total of 43 women with OC limited to one ovary (26 with borderline malignant tumors and 17 with invasive tumor FIGO IA) underwent surgical sparing therapy Disease recurrence rate as the measure of the quality of treatment and the number of pregnancies, together with their course, as the measure of the effectiveness of the fertility-sparing therapy were evaluated. RESULTS: The observation period, from 4-10 years for borderline malignancy and 3-8 years for invasive FIGO IA tumor revealed no cases of disease recurrence. In the former group, 14 (73 %) women conceived, out of whom 2 miscarried before 22 weeks of gestation, 1 give birth pre-term, i.e. at 34 weeks of gestation, 10 (71.5%) gave birth at term, and 1 is pregnant (6 months) at the time of the publication of the study In the later group, 9 (81.8%) women conceived, out of whom 1 miscarried at 10 weeks of gestation, 1 had a case of intrauterine fetal demise at 27 weeks of gestation, 6 gave birth at term, and 1 is pregnant (3 months) at the time of the publication of the study CONCLUSIONS: Proper patients selection for fertility-sparing management, as well as the way of performing the surgery allow for adequate control of the malignant process in patients with tumor limited to one ovary and preservation of fertility

[Conization and radical vaginal trachelectomy with laparoscopic lymphadenectomy in fertility-sparing surgical treatment of cervical cancer]. / Konizacja i radykalna pochwowa trachelektomia z laparoskopowa limfadenektomia w leczeniu chirurgicznym kobiet z rakiem szyjki macicy pozwalajacym na zachowanie plodnosci.

INTRODUCTION: In the mid-80's and 90's of the last century uterine-sparing surgical treatment methods were proposed to women wishing to preserve their fertility. OBJECTIVES: The aim of the study was to assess practical application of conization or radical vaginal trachelectomy with laparoscopic lymphadenectomy in women with squamous cell cervical cancer (FIGO stages IA and IB1) who want to retain their ability to procreate. Material and methods: A total of 119 women (aged 25-43 years) were included in the study Ninety-six women (60 - IA1 and 36 - IA2) were deemed eligible for conization and 23 women with stages IA2 and IB1 with neoplastic changes of <2cm in diameter were qualified for radical vaginal trachelectomy with laparoscopic lymphadenectomy RESULTS: Conization was not radical in 9 cases and these women were reoperated. A 5-year follow-up in patients after conization revealed disease recurrence (CIN) in 3 IA1 cases (6.5%), and 2 IA2 cases (9.1%). In patients with follow-up of <5 years, no recurrence was observed in 9 out of 10 A1 cases. Similarly there was no recurrence in 7 out of 9 lA2 cases. A 5-year follow-up in patients after trachelectomy confirmed disease recurrence (CIN) in 1 IA2 case (6.25%). There was no disease recurrence in 2 IA1 cases and 4 IB1 cases. Out of 8 subjects staged 1A2 with a follow-up of <5 years, no disease recurrence was observed in 7 (87.5%) patients. During the same follow-up time, no disease recurrence was detected in 3 patients staged IB1. CONCLUSIONS: Proper uterine-sparing treatment with appropriate qualification guarantees adequate control of the treatment process in women with cervical cancer stages IA and IB1 (<2 cm in diameter) according to FIGO.

Soluble TNF-α receptor I encoded on plasmid vector and its application in experimental gene therapy of radiation-induced lung fibrosis.

Post-radiation inflammatory reaction leads to an irreversible pulmonary fibrosis which may cause lethal respiratory insufficiency. Pathological inflammatory and fibrotic changes might be attenuated by inhibiting tumour necrosis factor (TNF)-α activity using TNF-α soluble receptors. Thus, an experimental antifibrotic gene therapy with the plasmid vector encoding a mouse soluble receptor I for TNF-α (psTNFR-I) was assessed. Soluble TNFR-I encoding gene was cloned into pcDNA3.1 plasmid. The ability of psTNFR-I expressing vector to transfect cells, and its biological activity in vitro and in vivo were examined by PCR, RT-PCR, MTT assay and ELISA. The C57Bl/6J mice received single intramuscular injection of psTNFR-I, conjugated with polyetylenimine (PEI) 25 kDa, equally divided to both hind legs, 3 days before irradiation (20 Gy, Co60), and either a single injection or ten injections once a week after irradiation. The data proved the effectiveness of psTNFR-I product to neutralise TNF-α activity in vitro. The in vivo plasmid incorporation and maintenance was confirmed. Measurements of plasma soluble TNFR-I levels showed that the in vivo gene transfer was effective. PEI was found to enhance transfection efficiency in vivo. The psTNFR-I/PEI complexes caused no toxicity in the transfected mice. C57Bl/6J mice that received prolonged psTNFR-I/PEI injections developed lethal fibrotic syndrome and died 8 weeks later than the mice treated with a double plasmid injection and the control mice treated with a control plasmid. Sequential administration of soluble TNFR-I by a nonviral, intramuscular gene transduction in the early and late post-radiation inflammatory phase prolonged survival of irradiated mice and attenuated the symptoms of lung fibrosis. The psTNFR-I gene transduction may provide a safe and simple method to partially neutralise TNF-α activity and prevent radiation-induced lung injury.

Cytokines of the Th1 and Th2 type in sera of rheumatoid arthritis patients; correlations with anti-Hsp40 immune response and diagnostic markers.

Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disease which affects approximately 1% of the population worldwide. Recent research on the role of heat shock proteins (Hsps) in RA development indicates that they may have pro- or anti-inflammatory effect, most probably via modulating cytokine secretion. We investigated type Th1 (INFγ, TNFα, IL-2) and type Th2 (IL-10, IL-6, IL-4) cytokine levels in sera of RA patients and healthy controls, using flow cytometric bead array assay, and searched for correlations between the cytokine levels and serum antibodies against bacterial (DnaJ) and human (Hdj1, Hdj2 and Hdj3) Hsp40 proteins, as well as clinical and laboratory parameters. The levels of all cytokines studied were significantly increased in RA patients; the highest increase relative to healthy controls (7-fold) was observed for IL-6 and its levels correlated positively with the antibodies directed to DnaJ and to the C-terminal domain of Hdj2, and with diagnostic parameters (DAS 28, Steinbrocker RTG criteria, ARA/7, ESR, TEN, SW and GH). INFγ levels correlated negatively with DAS 28, ESR, TEN and SW. No correlations were found for TNFα, IL-2 or IL-4. Our results support the hypothesis of Hsp40 involvement in RA as well as indicate that IL-6 serum level is a good marker of the RA activity.

The analysis of CD34 antigen immunoreactivity level in invasive ductal breast cancer with respect to the presence of lymph node metastases.

BACKGROUND: Angiogenesis is a basic process that enables neoplasms to thrive. Microvessel density (MVD) evaluation is an accepted parameter for assessing the angiogenesis process within a tumor. The aim of the present study has been to assess the number of microcapillaries in both invasive ductal breast cancer with the presence of metastases in regional lymph nodes and in invasive ductal breast cancer without such metastases. METHODS: The CD34 antigen immunoreactivity level was assessed by immunohistochemistry in both types of invasive ductal breast cancer. Tissue samples were obtained from 40 patients and were divided into two groups according to whether or not there were lymph node metastases. RESULTS: The patients with lymph node metastases exhibited statistically significantly higher numbers of stained microcapillaries than the patients who did not have lymph node metastases. CONCLUSION: Thus the number of stained microcapillaries as evaluated by using the CD34 immunoreactivity level seems to be a useful predictor for the development of local lymph node metastases in female invasive ductal breast cancer.

The DnaK chaperones from the archaeon Methanosarcina mazei and the bacterium Escherichia coli have different substrate specificities.

Hsp70 (DnaK) is a highly conserved molecular chaperone present in bacteria, eukaryotes, and some archaea. In a previous work we demonstrated that DnaK from the archaeon Methanosarcina mazei (DnaK(Mm)) and the DnaK from the bacterium Escherichia coli (DnaK(Ec)) were functionally similar when assayed in vitro but DnaK(Mm) failed to substitute for DnaK(Ec) in vivo. Searching for the molecular basis of the observed DnaK species specificity we compared substrate binding by DnaK(Mm) and DnaK(Ec). DnaK(Mm) showed a lower affinity for the model peptide (a-CALLQSRLLS) compared to DnaK(Ec). Furthermore, it was unable to negatively regulate the E. coli sigma32 transcription factor level under heat shock conditions and poorly bound purified sigma32, which is a native substrate of DnaK(Ec). These observations taken together indicate differences in substrate specificity of archaeal and bacterial DnaKs. Structural modeling of DnaK(Mm) showed some structural differences in the substrate-binding domains of DnaK(Mm) and DnaK(Ec), which may be responsible, at least partially, for the differences in peptide binding. Size-exclusion chromatography and native gel electrophoresis revealed that DnaK(Mm) was found preferably in high molecular mass oligomeric forms, contrary to DnaK(Ec). Oligomers of DnaK(Mm) could be dissociated in the presence of ATP and a substrate (peptide) but not ADP, which may suggest that monomer is the active form of DnaK(Mm).