Retroperitoneal myolipoma

BACKGROUND: Myolipoma is a benign tumour in which smooth muscle cells are mixed with adipocytes. CASE PRESENTATION: A 34-year old lady presented with a mass in the right iliac fossa detected on computerised tomographic (CT) scan. Wide excision of the retroperitoneal mass was done. Histopathology showed features of myolipoma. There was no recurrence or metastasis at three years. CONCLUSION: Myolipoma is a rare benign entity; hence a benign course and good prognosis are expected

Selective CDK9 inhibition overcomes TRAIL resistance by concomitant suppression of cFlip and Mcl-1.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can induce apoptosis in many cancer cells without causing toxicity in vivo. However, to date, TRAIL-receptor agonists have only shown limited therapeutic benefit in clinical trials. This can, most likely, be attributed to the fact that 50% of all cancer cell lines and most primary human cancers are TRAIL resistant. Consequently, future TRAIL-based therapies will require the addition of sensitizing agents that remove crucial blocks in the TRAIL apoptosis pathway. Here, we identify PIK-75, a small molecule inhibitor of the p110α isoform of phosphoinositide-3 kinase (PI3K), as an exceptionally potent TRAIL apoptosis sensitizer. Surprisingly, PI3K inhibition was not responsible for this activity. A kinome-wide in vitro screen revealed that PIK-75 strongly inhibits a panel of 27 kinases in addition to p110α. Within this panel, we identified cyclin-dependent kinase 9 (CDK9) as responsible for TRAIL resistance of cancer cells. Combination of CDK9 inhibition with TRAIL effectively induced apoptosis even in highly TRAIL-resistant cancer cells. Mechanistically, CDK9 inhibition resulted in downregulation of cellular FLICE-like inhibitory protein (cFlip) and Mcl-1 at both the mRNA and protein levels. Concomitant cFlip and Mcl-1 downregulation was required and sufficient for TRAIL sensitization by CDK9 inhibition. When evaluating cancer selectivity of TRAIL combined with SNS-032, the most selective and clinically used inhibitor of CDK9, we found that a panel of mostly TRAIL-resistant non-small cell lung cancer cell lines was readily killed, even at low concentrations of TRAIL. Primary human hepatocytes did not succumb to the same treatment regime, defining a therapeutic window. Importantly, TRAIL in combination with SNS-032 eradicated established, orthotopic lung cancer xenografts in vivo. Based on the high potency of CDK9 inhibition as a cancer cell-selective TRAIL-sensitizing strategy, we envisage the development of new, highly effective cancer therapies.Cell Death and Differentiation advance online publication, 20 December 2013; doi:10.1038/cdd.2013.179

CpG-island methylation study of liver fluke-related cholangiocarcinoma

Background: Genetic changes have been widely reported in association with cholangiocarcinoma (CCA), while epigenetic changes are poorly characterised. We aimed to further evaluate CpG-island hypermethylation in CCA at candidate loci, which may have potential as diagnostic or prognostic biomarkers. Methods: We analysed methylation of 26 CpG-islands in 102 liver fluke related-CCA and 29 adjacent normal samples using methylation-specific PCR (MSP). Methylation of interest loci was confirmed using pyrosequencing and/or combined bisulfite restriction analysis, and protein expression by immunohistochemistry. Results: A number of CpG-islands (OPCML, SFRP1, HIC1, PTEN and DcR1) showed frequency of hypermethylation in >28% of CCA, but not adjacent normal tissues. The results showed that 91% of CCA were methylated in at least one CpG-island. The OPCML was the most frequently methylated locus (72.5%) and was more frequently methylated in less differentiated CCA. Patients with methylated DcR1 had significantly longer overall survival (Median; 41.7 vs 21.7 weeks, P=0.027). Low-protein expression was found in >70% of CCA with methylation of OPCML or DcR1. Conclusion: Aberrant hypermethylation of certain loci is a common event in liver fluke-related CCA and may potentially contribute to cholangiocarcinogenesis. The OPCML and DcR1 might serve as methylation biomarkers in CCA that can be readily examined by MSP

Prognostic Significance of Wnt-1, β-catenin and E-cadherin Expression in Advanced Colorectal Carcinoma

Wnt/β-catenin pathway plays an important role in initiation and progression of colorectal oncogenesis. The aim of this study was to determine expression and localization of E-cadherin, β-catenin and Wnt-1 proteins in colorectal tumors. Expression of β-catenin, E-cadherin and Wnt-1 was determined by immunohistochemistry on advanced colorectal cancers. Abnormal expression of E-cadherin, β-catenin, Wnt-1 was observed. Additionally, we revealed correlations between levels of studied proteins and histoclinical data. In multivariate analysis nuclear β-catenin, higher carcinoembryonic antigen serum level before treatment, female sex and tumor localized in colon or rectum were independent unfavorable prognostic factors. These findings support the hypothesis that Wnt/β-catenin pathway plays an important role in advanced colorectal carcinoma

CCNE1 and survival of patients with tubo-ovarian high-grade serous carcinoma: An Ovarian Tumor Tissue Analysis consortium study

BACKGROUND: Cyclin E1 (CCNE1) is a potential predictive marker and therapeutic target in tubo-ovarian high-grade serous carcinoma (HGSC). Smaller studies have revealed unfavorable associations for CCNE1 amplification and CCNE1 overexpression with survival, but to date no large-scale, histotype-specific validation has been performed. The hypothesis was that high-level amplification of CCNE1 and CCNE1 overexpression, as well as a combination of the two, are linked to shorter overall survival in HGSC. METHODS: Within the Ovarian Tumor Tissue Analysis consortium, amplification status and protein level in 3029 HGSC cases and mRNA expression in 2419 samples were investigated. RESULTS: High-level amplification (>8 copies by chromogenic in situ hybridization) was found in 8.6% of HGSC and overexpression (>60% with at least 5% demonstrating strong intensity by immunohistochemistry) was found in 22.4%. CCNE1 high-level amplification and overexpression both were linked to shorter overall survival in multivariate survival analysis adjusted for age and stage, with hazard stratification by study (hazard ratio [HR], 1.26; 95% CI, 1.08-1.47, p = .034, and HR, 1.18; 95% CI, 1.05-1.32, p = .015, respectively). This was also true for cases with combined high-level amplification/overexpression (HR, 1.26; 95% CI, 1.09-1.47, p = .033). CCNE1 mRNA expression was not associated with overall survival (HR, 1.00 per 1-SD increase; 95% CI, 0.94-1.06; p = .58). CCNE1 high-level amplification is mutually exclusive with the presence of germline BRCA1/2 pathogenic variants and shows an inverse association to RB1 loss. CONCLUSION: This study provides large-scale validation that CCNE1 high-level amplification is associated with shorter survival, supporting its utility as a prognostic biomarker in HGSC

A combination of the immunohistochemical markers CK7 and SATB2 is highly sensitive and specific for distinguishing primary ovarian mucinous tumors from colorectal and appendiceal metastases

This study is supported by research funds from Cancer Research Society of Canada (19319). NSM is supported by the NSW Ministry of Health and UNSW Sydney under the NSW Health PhD Scholarship Program, and the Translational Cancer Research Network, a translational cancer research center program funded by the Cancer Institute NSW. The Gynaecological Oncology Biobank at Westmead was funded by Cancer Institute NSW (12/RIG/1–17 and 15/RIG/1–16) and the National Health and Medical Research Council of Australia (ID310670, ID628903). FM is funded by University of Pittsburgh School of Medicine Dean's Faculty Advancement Award. The HOPE study is funded by: US National Cancer Institute (K07-CA80668, P50-CA159981, R01CA095023), US Army Medical Research and Materiel Command (DAMD17–02–1–0669) and NIH/National Center for Research Resources/General Clinical Research Center (MO1- RR000056). KS is funded by the Swedish Cancer foundation. The Generations Study thank Breast Cancer Now, the Institute of Cancer Research and Ovarian Cancer Action for support and funding. The ICR acknowledge NHS funding to the NIHR Biomedical Research Centre. Tissue samples for GER were provided by the tissue bank of the National Center for Tumor Diseases (NCT, Heidelberg, Germany) in accordance with the regulations of the tissue bank and the approval of the ethics committee of the University of Heidelberg. The Health Science Alliance (HSA) Biobank acknowledges the UNSW Biorepository, UNSW Sydney, Australia. We thank Shuhong Liu, Young Ou, and Deon Richards for immunohistochemical stains, and Thomas Kryton, BFA, digital imaging specialist for Alberta Public Lab for creating the figures. We especially thank all the study participants, health care staff and data providers internationally who have made this research possible

Behavioral Indicators of Ovarian Phase in the Dromedary She-camel

In beef and dairy cattle the estrus cycle affects endocrine, ovarian, and behavioral events and detection of estrus by behavioral parameters is well assessed (Roelofs et al 2010). Female dromedary camels are seasonal breeders and induced ovulators, in which four follicular phases have been distinguished: recruitment, growth, mature and regressing (Skidmore 2011). Currently, little data has been published about estrus behavior in the she- camel. Recent rapid development in using assisted reproductive techniques to enhance dromedary reproduction makes urgent the detection of estrus in this species. The aim of this study was to relate different reproductive phases with observed behaviors which she-camels showed in presence of a male, and to identify behavioral indicators for the mature phase of estrus. Twenty-four healthy adult non-pregnant and non-lactating females were used. Two days prior to initiating the study, all received 789 µg Cloprostenol (Estrumate®, Agro-pharm Inc., Canada) intramuscularly (i.m.). Each animal was examined once a week over a three week period and was checked by manual gynecological examination, and its reproductive system was ultrasonographically scanned. Camels bearing mature preovulatory follicles (diameter >13 mm, Ismail et al 2007) were treated i.m. with 20 µg gonadorelin (Fertagyl®, Intervet, Germany) to induce ovulation, and ensure the presence of corpora lutea in the subsequent week. Immediately following examination sessions, each female was freely exposed to a restrained bull, and its behaviors recorded using a video camera. The videos were analyzed though a scan sampling ethogram (States: looking at the male; looking outside; standing; walking; searching; male interaction; lying down; Events: sniffing; interaction with the male; urination; defecation; sound emission). A score for tail position ("tail score": 1= close to the vulva, 2= horizontal, 3= vertical) and for interest in the bull ("male time score": from 1 to 5; 1=less than 20% of observation period spent near the bull; 5= more than 80%), were recorded. Dromedary females were considered "in estrus" when there was at least one >13 mm pre-ovulatory follicle, and "not in estrus" when growing follicles or corpora lutea were detected (Basioni 2007). All data were analyzed using a General Linear Model procedure (SAS 2007) where the independent variable was the physiological status (estrus/not in estrus) and the behavioral data were dependant variables. The main significant effect of estrus reproductive status (Table 1) was the increased curiosity in the male, particularly, increased duration (P = 0.009) and frequency (P = 0.0004) in male interaction, and spending more time close to the male (P < 0.0001). From this data, it seems that the interest in the male may be a good behavioral indicator of mature phase in the she-camel: the dam looks for and interacts with the male, sniffs around more, spends more time in standing quite near the male and increases tendency to lie down in front of him. Increased curiosity, searching out, looking at, and standing close to the male are reported as estrus behavioral indicators in sheep (Banks 1964). Detection of mature ovarian phase by behavioral indicators to improve timing to perform mating or artificial insemination may have profound implication for enhanced fertility in the dromedary camel (Manjunatha et al 2012), thus our data is a first step in this new and emergent research area. Table 1. Effect of reproductive status (estrus/not estrus) on the duration (sec) and the frequency (n/15 min) of the studied behavioral states and events. Data are expressed as square mean ± SE Duration (sec) Looking at the male Looking outside Standing Walking Searching Male interaction Lying down Estrus 184.2±26.4 247.6±40.7A 204.5±25.8a 48.3±9.6 204.3±41.7 2.7±0.33A 13.0±6.9 Not Estrus 143.1±24.1 380.8±37.1B 118.7±22.9b 61.0±8.7 181.5±38.0 1.5±0.3B 2.4±0.5 Frequency (n/15min) Sniffing Interaction with male Urination Defecation Sound Emission Tail Score Male Time Score Estrus 3.6±0.53 12.4±1.5A 0.2±01 0.2±0.0 21.4±3.6 1.4±0.1 3.6±0.1A Not Estrus 2.4±0.4 4.6±1.3B 0.3±0.1 0.1±0.0 25.9±3.3 1.5±0.1 2.4±0.1B Means followed by different letters differ statistically: A,B; P < 0.01 and a,b; P < 0.05 Bank, E.M. (1964). Behaviour 23, 249. Basioni, G.F. (2007). J. Biol. Sci. 7, 1038. Ismail, S.T., Al-Eknah, M-M., Hemeida N.A. (2007). J King Saud Uni 8, 51. Roelofs, J., López-Gatiusc, F., Hunterd, R.H.F., van Eerdenburge, F.J.C.M. et al. (2010). Theriogenology 74, 327. Manjunatha, B.M., Pratap, N., Al-Bulushi, S., Hago, B.E. (2012) Theriogenology 78, 965. Skidmore, J.A. (2011). Anim. Reprod. Sci. 124,148. This work was funded by European Union, PROCAMED Projec

Ramifications of protease-based liquefaction of camel semen on physical, kinematic and surface glyco-pattern of cryopreserved spermatozoa

The efficiency of incorporating different proteases in the diluent for reducing camel semen viscosity, and subsequent ramifications on morpho-functional and glycan surface properties of cryopreserved spermatozoa were investigated. Ejaculates (n = 48) were collected from three adult camels, Camelus dromedarius, during the breeding season (January - March). A portion of each raw ejaculate was evaluated for sperm physical and morphological traits, whereas the other portion was divided into three aliquots assigned for the following liquefaction treatments: control (untreated), 0.1 mg/mL papain or 5 U/mL bromelain. All samples were diluted with Tris-lactose diluent containing the anti-enzyme E-64 to neutralize both proteases before being processed for cryopreservation. Post-thaw physical and kinematic properties of spermatozoa were analyzed using a computer-assisted sperm analysis (CASA) system. The sperm surface glycocalyx pattern was evaluated with a panel of 14 fluorescent lectins. Although bromelain was more effective in elimination of semen viscosity, there was a negative correlation between bromelain supplementation and values for the variables: normal sperm, intact acrosome and intact sperm cell membrane. Bromelain supplementation, compared to papain-treated and control samples, was positively correlated with secondary sperm abnormalities, increased straight-line velocity (VSL, μm/s) and straightness (%) of spermatozoa. Results from the glycan analysis indicated that both proteases did not affect the N-linked glycan content of the entire sperm surface, whereas the treatment with proteases induced little change in N-acetylgalactosamine and fucose terminating glycans in the tail region of the sperm. Functional studies are needed to evaluate the sperm fertility rates of bromelain- and papain-treated semen for application in camel assisted reproductive technologies