Higher Levels Of Invasion Research Articles

ФЛОРА ТА РОСЛИННІСТЬ ІРПІНСЬКОГО ЛІСУ

It is reported about the diversity of phytobiota of the valuable natural complex "Irpin Forest" in the Kyiv region. The data on the history of research of this interesting from a botanical perspective territory of Kyiv Polissia are presented. The features of the vegetation of different complexes of this forest area, dominated by oak-pine, cereal and moss-lichen forests, are characterized. The most valuable forests here are oak ones, which are rich in rare herbaceous plant species. Besides, there fragmentary occur alder stands on lowland areas. The data on the flora of this territory, which includes 437 species of vascular plants from 67 families and 4 divisions, are presented. Ten dominating families of flora (Asreaceae, Poaceae Rosaceae, etc.) include 51.1 % of the forest flora species. A high level of adventization of this flora (19.6 %) has been noted, which is a result of the accessibility of the territory and a significant number of the population of adjacent settlements. Species with a high level of invasiveness include Amorpha fruticosa L., Iva xanthiifolia Nutt., Erigeron annuus (L.) Desf., Galinsoga parviflora Cav., Oenothera biennis L., Oxalis stricta L., Reynoutria japonica Houtt., Solidago canadensis L., Solanum nigrum L. They most often occur along forest roads, on meadows, in places of selective cuttings, on the fringes of the forest. On the periphery of the forest, a number of species were found which have fallen out of cultivation – Berberis aquifolium Pursh, Datura stramonium L. Gaillardia pulchella Foug., Ricinus communis L., Vitis vinifera L. The data on the rare plant species of this territory are given. There are 34 rare taxa, the most valuable of which are the representatives of the families Orchidaceae, Ranunculaceae, Poaceae. Five species of plants of the forest (Dactylorhiza fuchsii (Druce) Soó, D. incarnata (L.) Soó, Epipactis helleborine (L.) Crantz, Platanthera bifolia (L.) Rich., Pulsatilla patens (L.) Mill. Sl) are listed in the Red Data Book of Ukraine. It is proposed to create a new object of the nature reserve fund of the Kyiv region – a regional landscape park "Streams" ("Potoky") on a part of this territory, with further plans to create an administration, security service and develop a management plan for resource management of the object.

MicroRNA-33a-5p suppresses colorectal cancer cell growth by inhibiting MTHFD2.

Colorectal cancer (CRC) is one of the most common malignancies with high levels of invasiveness, drug resistance and mortality, but the internal mechanisms of CRC are largely unknown. MicroRNAs (miRs) have been reported to be involved in the development of CRC, and numerous studies have demonstrated that the abnormal expression of miR-33a-5p might be associated with CRC. However, the function and downstream mechanism of miR-33a-5p in colorectal cancer (CRC) remains unclear. Methylenetetrahydrofolate dehydrogenase 2 (MTHFD2), a mitochondrial enzyme involved in folic acid metabolism, interestingly was confirmed to be one of the target genes of miR-33a-5p in the present study. We first confirmed that miR-33a-5p in CRC tissues and cell lines were downregulated (P<0.05), and that the proliferation, clone formation capacities, G1/S progression, and migration capacities of the two CRC cell lines HCT116 cells and HT29 were suppressed by miR-33a-5p overexpression in vitro (P<0.05). Ctrl HCT116 and miR-33a-5p-overexpressing HCT116 were injected into nude mice. In vivo tumour formation was significantly suppressed by miR-33a-5p overexpression (P<0.05) as well as the proliferation marker Ki67 (P<0.05). Additionally, MTHFD2 protein expression was significantly enhanced in CRC tissues. From bioinformatics predictions and a luciferase report analysis, MTHFD2 was confirmed to be one of the target genes of miR-33a-5p. In contrast to the role of miR-33a-5p overexpression, MTHFD2 overexpression promoted the proliferation and migration of HCT116 and HT29 cells (P<0.05), which confirmed that MTHFD2 was a functional target gene of miR-33a-5p. In conclusion, miR-33a-5p inhibits the growth and migration of CRC by targeting MTHFD2.

Abstract 4875: Clinical importance and clarification of the WHO subclassification of combined hepatocellular and cholangiocarcinoma

Abstract Introduction: Combined hepatocellular and cholangiocarcinoma, classical type (Comb HCC-CCC) is defined as a single tumor, in which hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC) are clearly differentiated. However, combined hepatocellular-cholangiocarcinoma with stem-cell features, cholangiolocellular type (CoCC) was added to the WHO Classification in 2010. This has caused a great deal of confusion and raised many questions for clinicians and pathologists. The aim of this study was to answer these questions and evaluate the clinical importance of the WHO classification. Subjects: The subjects were 1,002 patients with primary liver cancer who underwent hepatectomy from 2010 to 2016, including 931 with HCC, 6 with Comb HCC-CCC, 50 with ICC, and 15 with CoCC. Pathological evaluation using immunohistochemical staining was mainly used for CoCC. Results: The CoCC cases had similar serum total bilirubin and serum albumin to those in other types of hepatic cancer, but with a low ICGR15 level. AFP, PIVKA-II, and CEA were 4.4 (1.3 - 3,689) ng/mL, 18 (9 - 15,933) mAU/mL, and 3.8 (0.2 - 33) ng/mL, respectively. In preoperative CT diagnosis of CoCC, HCC, Comb HCC-CCC, and ICC were diagnosed in 20%, 40%, and 40% of the cases, respectively, suggesting difficulty with this diagnosis. The resected specimen showed the number of tumors (1 [1 - 16]), tumor diameter (4 [1.7 - 12] cm), microscopic-portal venous invasion (43%), microscopic venous invasion (36%), and cirrhosis (22%), suggesting a higher level of invasion to vascular channels compared to other types of cancer. In pathological findings, both transitional HCC and cholangiocarcinoma were found in HE staining, in addition to fibrosis and necrosis in the tumor. Among the subjects, 40%, 100%, 100%, and 100% had biliary cell markers of positive CD117 (c-Kit), positive CD56 (NCAM), positive CK7, and positive CK19, respectively, in addition to high expression of neural cell adhesion molecules. Levels of hepatocytes (10%) and glypican 3 (25%) were low. Cumulative survival was lower than that in other types of liver cancer, with 1- and 3-year survival rates of 67% and 57%, respectively. [Conclusion] The development mechanism of Comb HCC-CCC is thought to be genetic transformation of HCC or ICC, while that of CoCC with stem-cell features is believed to be cancerization of stem cells. However, stem-cell features are just characteristics, and thus do not serve as definitive evidence for stem cell-derived cancer. In addition, since there are various types of CoCC in Comb HCC-CCC, actual classification is extremely difficult in many cases. The clinical importance of clear differentiation between Comb HCC-CCC and CoCC is unclear, and thus the WHO classification of these tumors may be in a transition period. Citation Format: Hisashi NAKAYAMA, Tadatoshi TAKAYAMA, Yutaka Midorikawa, Masahiko Sugitani. Clinical importance and clarification of the WHO subclassification of combined hepatocellular and cholangiocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4875.

Make it simpler: Alien species decrease functional diversity of coastal plant communities

AbstractQuestions(a) Are there differences in abundance‐weighted functional trait values between native and alien species in coastal plant communities? (b) Which functional traits are associated with a higher level of invasion in these communities? (c) Do functional diversity patterns differ between native and alien species? (d) Is alien species occurrence linked to small‐scale functional homogenization effects on the resident native species?LocationN‐Adriatic coastal ecosystems (Marano and Grado lagoon, Friuli Venezia Giulia region, Italy).MethodsWe sampled coastal vegetation within two habitats (foredunes and saltmarshes) along nine belt transects in two sampling sites. Plant species richness and abundance were assessed in 128 plots along with a suite of plant functional traits. We tested for differences in community‐weighted means between native and alien species within the two habitats, and Linear Mixed Models provided insights into traits fostering the invasion success among alien species. To check for potential functional homogenization driven by alien species invasion, we explored functional diversity patterns of native and alien species (β and γ functional diversity) and the relationship between α functional diversity and alien cover.ResultsAlien species had lower functional diversity than natives and were characterized by lower leaf construction costs coupled with lower drought resistance and higher water transport efficiency. The most abundant aliens were the ones minimizing carbon investment for leaf construction. In addition, we also found evidence for small‐scale functional homogenization driven by alien invasion.ConclusionsOur results suggested that native species adopt a resource conservative strategy whereas alien species are characterized by a higher resource acquisition capacity (i.e., acquisitive strategy). Our data also confirmed that alien species are less functionally diverse than natives, potentially driving the community towards small‐scale functional homogenization, resulting in a loss of species and a reduction in the functional space.

Multispecies plant invasion increases function but reduces variability across an understorey metacommunity

Several plant traits have been linked to invasion success in studies involving single alien species or invaded versus non-invaded communities. Less consideration has been given to how invasion by multiple alien species changes community-wide traits and functional structure at landscape scales. Changes to community functional structure by multispecies invasion may have consequences for ecosystem function. Therefore, it is important to identify which traits of naturalized aliens are responsible for optimizing patterns of trait convergence and divergence. Here, we examine if invasion by multiple alien species is related to changes in community-wide functional traits, and patterns of trait convergence and divergence, across an invasion gradient. We collected field data in Victoria, Australia, on traits previously associated with invasion success from 15 × 500 m2 plots. These plots encompassed a range of relative alien cover (~ 3–61%) and represented a multispecies invasion gradient. We tested relationships of each trait to the invasion gradient. We also determined trait convergence and divergence patterns across the gradient, using a trait divergence analysis. Specific leaf area and proportion of annual species significantly increased with the level of invasion. Annual life history and flowering duration maximised trait-convergence, but the two traits had different relationships to the invasion gradient. Specific leaf area, annual life history, leaf dry mass and seed mass maximised trait divergence. Evidence of trait-divergence along the gradient is represented by an increase in functional diversity with invasion level. However, variation in functional diversity was higher in little-invaded communities and stabilised at intermediate and high levels of invasion. Traits (specific leaf area and annual life history) associated with invasion success at a landscape scale are traits promoting rapid life cycle completion, and increases in multispecies invasion leads to an increase in functional diversity but a decline in its variation.

Isolation and Identification of Human Pathogenic Bacteria and Related Toxicity Potential to Different Cell Line

Objectives : The present study was undertaken to investigate three different virulence mechanisms (adherence, invasion and cytotoxicity) of isolated Escherichia coli from urine and stool specimens and the relation between these different mechanisms and the ability of isolated Escherichia coli to induce apoptosis in Vero cells. Methodology: Different clinical samples have been collected from some Egyptian hospitals ( Suez canal authority hospital, Benha university hospital and Ismailia university hospital ). Bacterial strains were isolated and identified by morphological and biochemical analysis using biolog plates for selecting Escherichia coli strains from clinical samples which resulted in 30 Escherichia coli strains. The In-vitro antimicrobial sensitivity of the E.coli isolates were tested against 11 antimicrobial drugs used in human treatment. Results: Five E.coli isolates were selected from five groups of E.coli isolates which were classified according to antibiotic sensitivity similarity. These five selected Escherichia coli isolates were screened for adherence after 1h , invasion after 3h , 5h and cytotoxicity against Vero cells. All the tested Escherichia coli isolates were able to adhere to Vero cells by variable degrees. Concerning invasion, the tested Escherichia coli isolates were found to have the highest levels of invasion reaching at 3 h and the invasion deacresed after 5h of this assay. on screening for cytotoxicity by the MTT assay, the tested Escherichia coli isolates with highly adherence and invasion showed highly cytotoxic effects to Vero cells after 6 h indicating a relation between adherence and cytotoxicity. using Annexin V/PI staining by flow cytometry, highly cytotoxic tested Escherichia coli isolates showed high percentage of apoptotic Vero cells . Upon screening for activation of caspase 3, highly cytotoxic strains showed highly activation in caspase 3 which plays a central central role in driving the apoptotic pathways indicating a relation between cytotoxic effect and induction apoptosis in Vero cells. Conclusions: These results suggest that the tested Escherichia coli strains exhibit different capacity to adhere to Vero cells and invasion is a post adherence effect and it's preferable time is 3 h because after this peroid Vero cells were ruptured. The cytotoxicity of the isolated Escherichia coli is associated with higher adherence and invasion to Vero cells. This demonstrates that Escherichia coli strains induce apoptosis of Vero cells at different degrees . Induction of apoptosis in Vero cells is associated with higher adherence ,invasion and cytotoxicity Escherichia coli of to Vero cells. The induction of apoptosis is correlated with the virulence of Escherichia coli.

Factors Influencing Disease Progression in Patients with Head and Neck Melanoma.

Histological parameters as well as the status of sentinel lymph node are known to be strong prognostic factors in patients with melanoma. In this study, we retrospectively analyzed 1,384 patients who were diagnosed with head and neck melanoma between 1976 and 2010 regarding prognostic factors [tumor thickness, level of invasion, sentinel lymph node (SLN) status, ulceration, histological subtype, localization, and gender], overall survival, and disease-free survival. Patients who developed metastases had a significantly thicker tumor than patients without metastases. Additionally, a thicker tumor was often associated with a higher level of invasion (Clark level). There was no overall survival benefit in patients who underwent SLN dissection when compared to patients who did not (p=0.07). Compared to SLN-negative patients, patients with SLN involvement had a significantly shorter disease-free period (p<0.001) and shorter overall survival time (p<0.001). In summary, tumor thickness is the most important prognostic factor. SLN dissection does not affect the overall survival of patients with melanoma. However, a positive SLN is a marker for a worse outcome in these patients.

Numerical Investigation of Four Seismic Upgrading Techniques on a Masonry Bell Tower

This paper presents some preliminary results from the numerical investigations of different interventions for the seismic upgrading of a masonry bell tower. Masonry bell towers belong to the cultural and historical heritage, are very widespread in Italy and their conservation has been given special attention in recent years. Their high vulnerability is related to the combination of mechanical material properties, geometrical irregularities, presence of inclination, foundation settlements and seismicity of the area. The present tower is located in Emilia Romagna Region, Northern Italy, recently stricken by a strong seismic sequence in May 2012. Analyses results show that the retrofitting of masonry bell towers is highly recommended in order to mitigate their present seismic risk. Four different upgrading techniques, consisting of grouted mortar, FRP sheets, internal steel frame and prestressing vertical rods, are investigated and compared in terms of structural performance improvement. The grouted mortar and internal steel frame techniques are found to be reasonable solutions when compared to the other proposals for the specific seismic intensity level. Furthermore, the other techniques have a relatively high level of invasiveness which is recommended to be avoid where possible. Experience has shown that the most effective interventions are those that not only protect the structure, but also preserve the built heritage.

The possible prognostic role of histone deacetylase and transforming growth factor β/Smad signaling in high grade gliomas treated by radio-chemotherapy: a preliminary immunohistochemical study.

Glioblastoma (GBM) is the most common and aggressive tumor of the central nervous system. Unfortunately, patients affected by this disease have a very poor prognosis, due to high level of invasiveness and resistance to standard therapies. Although the molecular profile of GBM has been extensively investigated, the events responsible for its pathogenesis and progression remain largely unknown. Histone Deacetylases (HDAC) dependent epigenetic modifications and transforming growth factor (TGF)-β/Smad pathway seem to play an important role in GBM tumorigenesis, resistance to common therapies and poor clinical outcome. The aim of this study was to evaluate the involvement and the possible interaction between these two molecular cascades in the pathogenesis and prognosis of GBM. Immunohistochemistry (IHC) was performed on microdissected GBM samples, collected from 14 patients (6 men and 8 women) ranging in age from 43 to 74 years. The patients were previously divided, on the basis of their overall survival (OS), into two groups: short and long OS. Patients with poor prognosis showed hyperexpression of HDAC4 and HDAC6, an activation of the TGF-β/Smad pathway, with high levels of IL-13, SMAD2, PDGF and MMP3 expression, compared to the long survivors. The short OS group exhibits a decrease in SMAD7 expression and also low levels of p21 immunostaining, which represents a common target of the two pathways. The IHC data was confirmed by quantitative analysis and Immunoblotting. Our preliminary results suggest that both HDAC4 and HDAC6 together with the TGF-β/Smad pathway may be involved in progression of GBM and this cross talking could be a useful prognostic marker in this deadly disease.

A conditionally replicating adenovirus expressing IL-24 acts synergistically with temozolomide to enhance apoptosis in melanoma cells in vitro.

Malignant melanoma is characterized by suppressed apoptosis in tumor cells and high levels of invasion. Temozolomide (TMZ) is one of the most effective single chemotherapeutic agents for patients with malignant melanoma, but resistance develops quickly and frequently. Therapeutic cytokines such as interleukin-24 (IL-24) and conditionally replicating adenoviruses have exhibited promising results as complementary therapies. Thus, the present study hypothesized that a conditionally replicating adenovirus expressing IL-24 combined with TMZ may exhibit increased antitumor activity compared with either treatment alone in melanoma A375 and M14 cell lines in vitro. The present study constructed an E1B-55 gene-deleted conditionally replicating adenovirus expressing the IL-24 gene (ZD55-IL-24). IL-24 was expressed at high levels in melanoma cells infected with ZD55-IL-24 in the presence of TMZ. The combination of ZD55-IL-24 + TMZ induced higher protein expression levels of the proapoptotic proteins B-cell lymphoma-2 (Bcl-2)-like protein 4 and phosphorylated protein, γ-H2A histone family member X (γ-H2AX), and reduced the levels of the antiapoptotic proteins Bcl-2, myeloid cell leukemia-1and nuclear factor-κB compared with either treatment individually. A dose-dependent increase in the cytopathic effects for the combination of ZD55-IL-24 and TMZ was also observed. The data of the present study suggest that the ZD55-IL-24 + TMZ combination induced increased levels of apoptosis, possibly by triggering DNA damage, in melanoma cells in vitro compared with either treatment alone. These findings suggest that this strategy may be a promising approach for the treatment of patients with malignant melanoma.

Metabolic determinants in Listeria monocytogenes anaerobic listeriolysin O production

Listeria monocytogenes is a human pathogen and a facultative anaerobe. To better understand how anaerobic growth affects L. monocytogenes pathogenesis, we first showed that anaerobic growth led to decreased growth and changes in surface morphology. Moreover, compared to aerobically grown bacteria, anaerobically grown L. monocytogenes established higher level of invasion but decreased intracellular growth and actin polymerization in cultured cells. The production of listeriolysin O (LLO) was significantly lower in anaerobic cultures—a phenotype observed in wild type and isogenic mutants lacking transcriptional regulators SigB or CodY or harboring a constitutively active PrfA. To explore potential regulatory mechanisms, we established that the addition of central carbon metabolism intermediates, such as acetate, citrate, fumarate, pyruvate, lactate, and succinate, led to an increase in LLO activity in the anaerobic culture supernatant. These results highlight the regulatory role of central carbon metabolism in L. monocytogenes pathogenesis under anaerobic conditions.

Post-Chornobyl Papillary Thyroid Carcinoma

Recent studies have confirmed that an increase in the number of patients with papillary thyroid cancer is due to the effects of the Chornobyl accident, and a cohort of persons who lived in the areas being exposed to radioactive contamination in 1986 has an increased risk of thyroid carcinoma even 30 years after the disaster.The objective of the research was to evaluate the results of treatment as well as to determine an optimal protocol of diagnosis, therapy and monitoring of patients with thyroid papillary carcinoma developed among the population of Ukraine in the period after the Chornobyl disaster.Materіals and methods. The analysis of treatment of 6,239 patients with papillary thyroid cancer during 1990-2015 was made. Follow-up period lasted from 1 to 25 years after initial surgery, on average 11.8±2.1 years. The age of patients ranged from 7 to 74 years. The mean age was 38.3±7.4 years. There were 5,003 (80.2%) females and 1,236 (19.8%) males. Results. Papillary thyroid carcinomas of the early period of the accident were characterized by a short latency period, high biological potential of malignancy with high level of invasiveness (extrathyroidal invasion in 51.7%; regional lymphatic metastases in 36.7%; distant metastases to the lungs in 5.2%). Over time, an improvement in the quality of ultrasound and cytological diagnosis allowed increasing the number of surgeries performed at the early stages of carcinoma development (up to 59.8% of cases). All patients underwent surgical treatment: thyroidectomy (84.6%), neck dissection of different lengths (27.8%), hemithyroidectomy in case of low-risk minimally invasive carcinoma (15.4%). In the group of pediatric patients at the time of the Chornobyl accident, the cumulative 15-year survival rate according to Kaplan-Meier method was 98.9%.Conclusions. In case of preoperative cytologic diagnosis of “thyroid carcinoma” in patients who were children at the time of the Chornobyl accident, it is mandatory to perform total thyroidectomy in combination with preventive central neck dissection (level VI lymph nodes) regardless of the degree of tumor spread. Modified lateral neck dissection is indicated in case of confirmation of metastases. Further ablative radioiodine therapy allows evaluating the effectiveness of treatment according to the level of serum thyroglobulin and antibodies thereto, and early detection and surgical removal of iodine-refractory metastases does not affect survival rates.

Landscape structure and climate affect plant invasion in subtropical grasslands

AbstractQuestionHow do climate and landscape structure affect plant invasion process in south Brazilian natural grasslands?LocationNatural grasslands across Rio Grande do Sul state, southern Brazil.MethodsWe selected 20 plots (2 km × 2 km each), situated in ten different grassland types in southern Brazil. In each grassland type, one plot was located in a region with remnant grassland cover &lt;30% (not conserved landscapes) and another in a region with remnant grassland cover &gt;60% (conserved landscapes), in a paired design. In each plot, we surveyed the presence and cover of the four most important invasive species of grasslands in Rio Grande do Sul (Cynodon dactylon, Eragrostis plana, Senecio madagascariensis and Ulex europaeus). We used 12 variables related to climate and landscape structure (i.e. composition and configuration) for testing causal models of plant invasion using Akaike's information criterion for path analyses.ResultsEmpirical evidence on how climatic conditions and landscape structure can cause plant invasion in subtropical grasslands was provided through six causal models validated with path analysis. The models indicated that increased water deficit, higher density of roads and losses in natural grassland cover caused an increment of invasive plant cover. Further, invasion differed between plots located in conserved vs not conserved landscapes, resulting in not conserved grasslands having a higher level of invasion. E. plana, a perennial grass from South Africa, was the most important invasive species in the grasslands, in terms of frequency and cover.ConclusionsOur results identified the general conditions that promote successful population growth of invasive plants in subtropical grasslands: water deficit, road density and land use, highlighting the effects of climate, landscape structure and human activity in the level of invasion across grassland types.

Invading a mutualistic network: to be or not to be similar.

Biological invasion remains a major threat to biodiversity in general and a disruptor to mutualistic interactions in particular. While a number of empirical studies have directly explored the role of invasion in mutualistic pollination networks, a clear picture is yet to emerge and a theoretical model for comprehension still lacking. Here, using an eco‐evolutionary model of bipartite mutualistic networks with trait‐mediated interactions, we explore invader trait, propagule pressure, and network features of recipient community that contribute importantly to the success and impact of an invasion. High level of invasiveness is observed when invader trait differs from those of the community average, and level of interaction generalization equals to that of the community average. Moreover, multiple introductions of invaders with declining propagules enhance invasiveness. Surprisingly, the most successful invader is not always the one having the biggest impact on the recipient community. The network structure of recipient community, such as nestedness and modularity, is not a primary indicator of its invasibility; rather, the invasibility is best correlated with measurements of network stability such as robustness, resilience, and disruptiveness (a measure of evolutionary instability). Our model encompasses more general scenarios than previously studied in predicting invasion success and impact in mutualistic networks, and our results highlight the need for coupling eco‐evolutionary processes to resolve the invasion dilemma.

Characterization of cytoplasmic cyclin D1 as a marker of invasiveness in cancer.

Cyclin D1 (Ccnd1) is a proto-oncogen amplified in many different cancers and nuclear accumulation of Ccnd1 is a characteristic of tumor cells. Ccnd1 activates the transcription of a large set of genes involved in cell cycle progress and proliferation. However, Ccnd1 also targets cytoplasmic proteins involved in the regulation of cell migration and invasion. In this work, we have analyzed by immunohistochemistry the localization of Ccnd1 in endometrial, breast, prostate and colon carcinomas with different types of invasion. The number of cells displaying membranous or cytoplasmic Ccnd1 was significantly higher in peripheral cells than in inner cells in both collective and pushing invasion patterns of endometrial carcinoma, and in collective invasion pattern of colon carcinoma. Also, the cytoplasmic localization of Ccnd1 was higher when tumors infiltrated as single cells, budding or small clusters of cells. To evaluate cytoplasmic function of cyclin D1, we have built a variant (Ccnd1-CAAX) that remains attached to the cell membrane therefore sequestering this cyclin in the cytoplasm. Tumor cells harboring Ccnd1-CAAX showed high levels of invasiveness and metastatic potential compared to those containing the wild type allele of Ccnd1. However, Ccnd1-CAAX expression did not alter proliferative rates of tumor cells. We hypothesize that the role of Ccnd1 in the cytoplasm is mainly associated with the invasive capability of tumor cells. Moreover, we propose that subcellular localization of Ccnd1 is an interesting guideline to measure cancer outcome.

Possible interactions between HDACs and TGF-beta/Smads pathway in Glioblastoma

Glioblastoma (GBM) is the most common and aggressive tumor of the Central Nervous System (CNS). Unfortunately, patients afflicted with this disease have a very poor prognosis, due to high level of invasiveness and resistance to standard therapies (1). Although the molecular profile of GBM has been extensively investigated, the events responsible for its pathogenesis and progression remain largely unknown. Reports have indicated that HDAC (Histone deacetylases) dependent epigenetic modifications (2) and the Tgfβ/Smad pathway (3) play roles in GBM tumorigenesis. The aim of this study was to evaluate the involvement and the possible interaction between these two molecular cascades in the pathogenesis, therapeutic responsiveness and prognosis of GBM. Immunohistochemestry (IHC) was performed on microdissected GBM samples, collected from 14 patients (n.8 men and n.6 women) ranging in age from 43 to 74 years. The patients were previously divided, on the basis of their overall survival (OS), into three groups: low, intermediate and high OS. Patients with poor prognosis showed hyperexpression of HDAC4 and HDAC6, an activation of the Tgfβ/Smad pathway, with high levels of IL-13, Smad2, PDGF and MMP3 expression, compared to the intermediate and high OS groups, whereas the expression of Smad7 was reduced. The high OS group also exhibits an increase in p21 immunostaining, which represents a common target of the two cascades. The IHC data was confirmed by Immunoblotting. Our results suggest that both HDAC4 and HDAC6 together with the Tgfβ/Smad pathway are involved in progression of GBM and could be a useful prognostic markers and may predict responsive to therapy.

Clinicopathological significance of orphan nuclear receptor Nurr1 expression in gastric cancer.

Gastric cancer is the fourth most common cancer and the second leading cause of cancer-related deaths worldwide. Gastric cancer is characterized by high levels of invasion and metastasis. Increasing attention is being focused on discovering molecular markers for the diagnosis of gastric cancer and for predicting its prognosis. The objective of the present study was to evaluate Nurr1 expression in gastric cancer and to assess its correlation with clinicopathological parameters and prognosis in gastric cancer patients. Tissue samples were obtained from 120 gastric cancer patients. We investigated Nurr1 expression in human normal and gastric cancer tissues using real-time reverse transcription polymerase chain reaction (qRT-PCR), western blotting, and immunohistochemistry. We determined the association between Nurr1 and recurrence, prognosis and patient clinicopathological parameters. Univariate and multivariate survival analyses with a Cox's proportional hazards regression model were used to identify independent factors related to recurrence and prognosis. The immunohistochemical, qRT-PCR and western blot analyses revealed that Nurr1 expression was increased in gastric cancer tissues compared with normal gastric tissue (P < 0.05). Nurr1 expression was significantly correlated with the tumor size, depth of tumor invasion, lymph node metastasis, recurrence, and distant metastasis of gastric cancer (P < 0.05). Moreover, Nurr1-high patients also exhibited poorer overall survival (OS) and disease-free survival compared with Nurr1-low patients (P < 0.01). The univariate and multivariate survival analyses suggested that Nurr1 expression (P = 0.011), histology (P = 0.018), depth of tumor invasion (P = 0.037), and presence of lymph node metastasis (P = 0.031) were independent prognostic factors for recurrence. In addition, Nurr1 expression (P = 0.007), depth of tumor invasion (P = 0.014), lymph node metastasis (P = 0.044), distant metastasis (P = 0.023), and recurrence (P = 0.011) were independent prognostic factors of OS in gastric cancer patients. The Nurr1 protein may be useful as a marker of recurrence, metastasis, and poor prognosis following curative resection in patients with gastric cancer.

Abstract 48: A novel method for live cell sorting based on RNA expression levels of cancer biomarkers maintains normal cellular function as confirmed by downstream functional testing

Abstract Enrichment of cells within a mixed population has been confined to fixed cells or in the case of live cell sorting to a limited set of cell surface markers which are typically not specific to the target population. Furthermore, cell surface antibodies may activate cell surface receptors altering the state of the cells such that they are no longer representative of the original population. Intracellular staining requires fixation and permeabilization and thus renders the cells unusable in downstream functional assays. Through the use of a unique nanoparticle sequence based detection technology we have shown the ability to detect specific RNA within live cells which does not affect or alter the cells and allows further testing of the target population following the enrichment. Both microRNA and mRNA species have been used to enrich for specific target populations through Fluorescence Activated Cell Sorting (FACS). Sorting cells based on the expression level of any specific RNA allows researchers the ability to utilize targets of biological interest within isolated populations of cells. Since this method is non-destructive to live cells the enriched cells can then be used for downstream functional assays. Here we present data showing the live cell enrichment of a target cell population within a heterogeneous mixture of cells based on their RNA expression levels. RNA probes to miR-221 and miR-222 were generated for the enrichment of invasive cells from the non-invasive. A mixture of breast cancer cells (MD-MB-231 and T47D) were sorted based on their expression levels of the RNA probes through FACS. The high and low expressing sort products were then each subjected to a collagen invasion assay. As expected the fractions which showed high expression of miR-221 and miR-222 showed a high level of invasion through the matrix while the low expressing fractions showed extremely low invasion. Live cell sorting based on RNA levels of cancer biomarkers in a non-destructive manner enables researchers the ability to study the expression of a diverse group of targets beyond the traditional surface markers with the added benefit of utilizing the enriched cells for downstream functional assays. Citation Format: Don Weldon, Victor Koong, Yuko Williams. A novel method for live cell sorting based on RNA expression levels of cancer biomarkers maintains normal cellular function as confirmed by downstream functional testing. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 48. doi:10.1158/1538-7445.AM2014-48

Population dynamics and reproductive biology ofAchatina fulica Bowdich, 1822 (Mollusca, Gastropoda) in Salvador - Bahia

The risks following introduction of invasive species are enormous and incalculable. Achatina fulica is considered one of the 100 species of greatest invasive potential, stands out among the land pulmonate snails, mostly for its high reproductive potential that accelerates the process of dispersion, increasing the damages related to health (disease transmission), to economy (crops destruction) and to environment (biodiversity loss). In order to investigate the relationship of the African snail with the environment, trying to relate sexual activity with climatic variables (temperature, rainfall and humidity), a study was conducted on aspects of population dynamics and reproductive biology in Salvador city. The snails were collected by hand, monthly. Morphometric data were obtained in the field and dissections to analyze the reproductive system at laboratory. To test if there was a greater sexual activity in wet periods was performed first a Principal Component Analysis (PCA) with climatic variables, followed by a regression between precipitation (the most influential environmental variable) and the variable of interest (sexual activity). The results showed that there is an apparent annual cycle forA. fulica, with a recruitment period covering the end of rainy season and the dry season (August to December 2006 and February 2007). Increase of shell size (height) and of sexual activity were observed during the rainy season, although snails were found in full sexual activity during the whole period of study. The von Bertalanffy curve showed that the oldest specimen collected would have three years and eleven months. The regression between precipitation and sexual activity was significant (P = 0.002) showing that the higher rainfall increased sexual activity. In addition, it was observed that there is an increase in the thickness of peristome as the specimens reaches sexual maturity however this relationship isn’t precise (it were found individuals with sexual structures not consistent with the reproductive stage given by the thickness of peristome) and should be set for each region studied. As the eradication becomes impossible because of high level of invasion found in Salvador, it is recommended to control the population of African snails by continuously removing specimens, especially in the rainy periods.

A non-surgical model of cervical spinal cord injury induced with focused ultrasound and microbubbles.

The most commonly used animal models of spinal cord injury (SCI) involve surgical exposure of the dorsal spinal cord followed by transection, contusion or compression. This high level of invasiveness often requires significant post-operative care and can limit post-operative imaging, as the surgical incision site can interfere with coil placement for magnetic resonance imaging (MRI) during the acute phase of SCI. While these models are considered to be similar to human SCI, they do not occur in a closed vertebral system as do the majority of human injuries. Here we describe a novel, non-surgical model of SCI in the rat using MR-guided focused ultrasound (FUS) in combination with intravenous injection of microbubbles, applied to the cervical spinal cord. The injury was well-tolerated and resulted in cervical spinal cord damage in 60% of the animals. The area of Gd-enhancement immediately post-FUS and area of signal abnormality at 24h were correlated with the degree of injury. The extent of injury was easily visualized with T2-weighted MRI and was confirmed using histology. Pathology was similar to that seen in other rat models of direct spinal cord contusion and compression. Unlike these methods, FUS is non-surgical and has lower mortality than seen in other models of cervical SCI. We developed a novel model of SCI which was non-surgical, well-tolerated, localized, and replicated the pathology seen in other models of SCI.