COX2 Effects on endometrial carcinomas progression.

Uterine corpus cancer is one of the most prevalent gynecologic malignancies, among which endometrial cancers (EC) represent about 90 %. Despite the proven predictive value of several immunohistochemical markers, there remains a need to identify new indicators of EC progression and exploit them for therapeutic purposes. Potential candidates with diagnostic and therapeutic efficacy include cyclooxygenases (COXs). We studied 50 EC cases: 30 endometrioid (EEC), 10 serous (SEC), 10 clear-cell endometrial carcinomas (CCEC) and 10 cases of normal endometrial tissues. We investigated the expression of COX2, ER, PR, Ki-67, EGFR, p53, Bcl-2, VEGF, MMP1, CD31, and CD163 immunohistochemically. COX2 levels in EC tissue are elevated compared to the normal endometrium and depend on tumour histological features and differentiation. Elevated COX2 leads to increased tumour cell proliferation, apoptosis inhibition, increased VEGF expression, microvessel density, and M2 macrophage infiltration, and inhibition of PR expression. ER, EGFR, and MMP1 levels are unaffected by COX2, whose levels are independent of patient age and FIGO stage.

How Useful is COVID-19 Antibody Testing - A Current Assessment for Oncologists.

The coronavirus disease 2019 (COVID-19) pandemic due to infection by a new human coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has seriously disrupted the provision of oncology services and their uptake. Antibody testing, both at an individual level and of populations, has been widely viewed to be a key activity for guiding the options for treatment of high-risk individuals, as well as the implementation of safe control of infection measures. Ideally, the detection of a specific antibody should signify that all individuals tested have been infected by SARS-CoV-2 and that in the case of specific IgG that they are immune to further infection. This would enable SARS-CoV-2-infected individuals to be appropriately managed and healthcare workers shown to be immune to return to work where they would no longer pose a risk to their patients or be at risk themselves. Unfortunately, this is not the case for COVID-19, where it has been shown that immunity may not be protective, and seroconversion delayed or absent. The variability in antibody test performance, particularly that of lateral flow assays, has caused confusion for the public and healthcare professions alike. Many antibody test devices have been made available without independent evaluations and these may lack both adequate sensitivity and specificity. This review seeks to educate healthcare workers, particularly those working in oncology, of the current benefits and limitations of SARS-CoV-2 antibody testing.

Making enhanced recovery the norm not the exception.

BACKGROUND: Enhanced Recovery After Surgery ("STAAR" in our system) is multimodal care focused on the reduction of physiological and psychological stress. While enhanced recovery is well established in colorectal surgery, and there is evidence for effectiveness in other surgical disciplines, to date widespread use is limited. METHOD: We implemented a Lean process that, within 12 months, expanded STAAR to 13 surgical services lines involving >130 surgeons, and impacting the care of >6000 surgical patients/year. RESULTS: Implementation involved educational and administrative meetings (279 in the first 6 months) and rounding. Use of STAAR was defined as >60% compliance. LOS was reduced up to 40%, mortality index and transfusion decreased 67% and 23% respectively. Case mix index increased 17%. Readmission rates, infections, ER visits were not increased. CONCLUSION: Using a Lean process focused on value, STAAR protocols became the standard rather than the exception. Time investment by senior surgical leadership was extensive.

Heavy metals effect on breast cancer progression.

BACKGROUND: Breast cancer is the most frequent localization of malignant process in American women and women of European countries. To date it is not possible to control the morbidity growth due to lack of effective ways of primary prevention. Comparing the incidence of breast cancer in developed countries with the countries of Asia and Africa, there is the fact of population predominance lesion in more urbanized countries. This suggests that the environment along with other factors, occupies a significant place in the initiation and progression of breast neoplasia. The impressive rates of industrial development led to the pollution of soil, surface water and, as a consequence, food by heavy metal salts.The purposes of this paper are as follows: the chemical composition determination of neoplastic breast tissue, evaluation of the DNA methylation level, study of prognostic-important receptors expression in the breast cancer cells, establishing linkages between all the derived indicators. METHODS: In our study we used the following methods: studying of the chemical composition of breast cancer tissue by atomic absorption spectrophotometry and energy-dispersion spectrometer; іmmunohistochemical study of ER, PR, HER2/neu, p53, Ki-67, E-cadherin and MGMT receptors; DNA extraction and investigation by oscillating infrared spectroscopy method. RESULTS: The total amount of heavy metals in breast cancer tissue ranged from 51.21 × 10-3 to 84.86 × 10-3 µg/kg. We have got the following results: the growth of heavy metals in neoplastic tissue is accompanied with the increase of HER2/neu, p53, Ki-67, MGMT expression and decrease of ER and PR expression. The increment of pathological DNA methylation is accompanied with the increasing amount of heavy metals in tumor tissue. CONCLUSIONS: Heavy metals through different pathogenetic links stimulate the progression of breast cancer and reduce its sensitivity to treatment. DNA of tumor tissue has a different level of methylation which changes with the amount of heavy metals in cancer cells. This is displayed on the synthesis of prognostically important receptors in neoplastic tissue.

Imprinted loci in domestic livestock species as epigenomic targets for artificial selection of complex traits.

The phenomenon of genomic imprinting, whereby a subset of mammalian genes display parent-of-origin-specific monoallelic expression, is one of the most active areas of epigenetics research. Over the past two decades, more than 100 imprinted mammalian genes have been identified, while considerable advances have been made in elucidating the molecular mechanisms governing imprinting. These studies have helped to unravel the epigenome--a separate layer of regulatory information contained in eukaryotic chromosomes that influences gene expression and phenotypes without involving changes to the underlying DNA sequence. Although most studies of genomic imprinting in mammals have focussed on mouse models or human biomedical disorders, there is burgeoning interest in the phenotypic effects of imprinted genes in domestic livestock species. In particular, research has focused on imprinted genes influencing foetal growth and development, which are associated with economically important production traits in cattle, sheep and pigs. These findings, when coupled with the data emerging from the various different livestock genome projects, have major implications for the future of animal breeding, health and management. Here, we review current scientific knowledge regarding genomic imprinting in livestock species and evaluate how this information can be used in modern livestock improvement programmes.

PHLDA2 is an imprinted gene in cattle.

Genomic imprinting is an epigenetic non-Mendelian phenomenon found predominantly in placental mammals. Imprinted genes display differential expression in the offspring depending on whether the gene is maternally or paternally inherited. Currently, some 100 imprinted genes have been reported in mammals, and while some of these genes are imprinted across most mammalian species, others have been shown to be imprinted in only a few species. The PHLDA2 gene that codes for a pleckstrin homology-like domain, family A (member 2), protein has to date been shown to be a maternally expressed imprinted gene in humans, mice and pigs. Genes subject to imprinting can have major effects on mammalian growth, development and disease. For instance, disruption of imprinted genes can lead to aberrant growth syndromes in cloned domestic mammals, and it has been demonstrated that PHLDA2 mRNA expression levels are aberrant in the placenta of somatic clones of cattle. In this study, we demonstrate that PHLDA2 is expressed across a range of cattle foetal tissues and stages and provide the first evidence that PHLDA2 is a monoallelically expressed imprinted gene in cattle foetal tissues, and also in the bovine placenta.

Single nucleotide polymorphisms in the imprinted bovine insulin-like growth factor 2 receptor gene (IGF2R) are associated with body size traits in Irish Holstein-Friesian cattle.

The regulation of the bioavailability of insulin-like growth factors (IGFs) is critical for normal mammalian growth and development. The imprinted insulin-like growth factor 2 receptor gene (IGF2R) encodes a transmembrane protein receptor that acts to sequester and degrade excess circulating insulin-like growth factor 2 (IGF-II) - a potent foetal mitogen - and is considered an important inhibitor of growth. Consequently, IGF2R may serve as a candidate gene underlying important growth- and body-related quantitative traits in domestic mammalian livestock. In this study, we have quantified genotype-phenotype associations between three previously validated intronic bovine IGF2R single nucleotide polymorphisms (SNPs) (IGF2R:g.64614T>C, IGF2R:g.65037T>C and IGF2R:g.86262C>T) and a range of performance traits in 848 progeny-tested Irish Holstein-Friesian artificial insemination sires. Notably, all three polymorphisms analysed were associated (P ≤ 0.05) with at least one of a number of performance traits related to animal body size: angularity, body depth, chest width, rump width, and animal stature. In addition, the C-to-T transition at the IGF2R:g.65037T>C polymorphism was positively associated with cow carcass weight and angularity. Correction for multiple testing resulted in the retention of two genotype-phenotype associations (animal stature and rump width). None of the SNPs analysed were associated with any of the milk traits examined. Analysis of pairwise r(2) measures of linkage disequilibrium between all three assayed SNPs ranged between 0.41 and 0.79, suggesting that some of the observed SNP associations with performance may be independent. To our knowledge, this is one of the first studies demonstrating associations between IGF2R polymorphisms and growth- and body-related traits in cattle. These results also support the increasing body of evidence that imprinted genes harbour polymorphisms that contribute to heritable variation in phenotypic traits in domestic livestock species.

RAGE-dependent regulation of calcium-binding proteins S100A8 and S100A9 in human THP-1.

Proinflammatory cell activation via the receptor for advanced glycation end products (RAGE) pathway may play a central pathogenetic role in atherosclerosis. Since S100A8/A9 was recently identified as ligand of RAGE, we determined the effects of proinflammatory cytokines on RAGE-mediated induction of gene expression of S100A8 and S100A9. mRNA levels of S100A8 and S100A9 were upregulated following cytokine stimulation with IL-6 (1, 10, 100 ng/ml) or TNFα (10 ng/ml) in human THP-1 cells. Preincubation of cells with 2000 ng/ml AGE (advanced glycation end products) before cytokine stimulation resulted in upregulation of RAGE. Pretreatment of THP-1 with AGE followed by stimulation with IL-6 (10 ng/ml) or TNFα (10 ng/ml) further increased S100A8 and S100A9 mRNA expression and S100A8/A9 release into cell culture supernatant, as compared to pretreatment with non-glycated albumin as control. Binding of AGE to RAGE was blocked with a neutralizing anti-RAGE antibody. Normal mouse IgG served as control. Cytokine-stimulated induction of S100A8 and S100A9 mRNA levels as well as of S100A8/A9 release after preincubation of cells with AGE were significantly suppressed by RAGE blockade, indicating a RAGE-dependent pathway of AGE-mediated S100A8/A9 expression.The cytokine-induced potentiated S100A8 and S100A9 expression under conditions with a high AGE burden is able to aggravate proinflammatory conditions via activation of the RAGE pathway.

High concordance of bovine single nucleotide polymorphism genotypes generated using two independent genotyping strategies.

Single nucleotide polymorphisms (SNPs) represent the most common form of DNA sequence variation in mammalian livestock genomes. While the past decade has witnessed major advances in SNP genotyping technologies, genotyping errors caused, in part, by the biochemistry underlying the genotyping platform used, can occur. These errors can distort project results and conclusions and can result in incorrect decisions in animal management and breeding programs; hence, SNP genotype calls must be accurate and reliable. In this study, 263 Bos spp. samples were genotyped commercially for a total of 16 SNPs. Of the total possible 4,208 SNP genotypes, 4,179 SNP genotypes were generated, yielding a genotype call rate of 99.31% (standard deviation ± 0.93%). Between 110 and 263 samples were subsequently re-genotyped by us for all 16 markers using a custom-designed SNP genotyping platform, and of the possible 3,819 genotypes a total of 3,768 genotypes were generated (98.70% genotype call rate, SD ± 1.89%). A total of 3,744 duplicate genotypes were generated for both genotyping platforms, and comparison of the genotype calls for both methods revealed 3,741 concordant SNP genotype call rates (99.92% SNP genotype concordance rate). These data indicate that both genotyping methods used can provide livestock geneticists with reliable, reproducible SNP genotypic data for in-depth statistical analysis.

A catalogue of validated single nucleotide polymorphisms in bovine orthologs of mammalian imprinted genes and associations with beef production traits.

Genetic (or 'genomic') imprinting, a feature of approximately 100 mammalian genes, results in monoallelic expression from one of the two parentally inherited chromosomes. To date, most studies have been directed on imprinted genes in murine or human models; however, there is burgeoning interest in the effects of imprinted genes in domestic livestock species. In particular, attention has focused on imprinted genes that influence foetal growth and development and that are associated with several economically important production traits in cattle, sheep and pigs. We have re-sequenced regions in 20 candidate bovine imprinted genes in order to validate single nucleotide polymorphisms (SNPs) that may influence important production traits in cattle. Putative SNPs detected via re-sequencing were subsequently re-formatted for high-throughput SNP genotyping in 185 cattle samples comprising 138 performance-tested European Bos taurus (all Limousin bulls), 29 African B. taurus and 18 Indian B. indicus samples. Analysis of the resulting genotypic data identified 117 validated SNPs. Preliminary genotype-phenotype association analyses using 83 SNPs that were polymorphic in the Limousin samples with minor allele frequencies ⩾0.05 revealed significant associations between two candidate bovine imprinted genes and a range of important beef production traits: average daily gain, average feed intake, live weight, feed conversion ratio, residual feed intake and residual gain. These genes were the Ras protein-specific guanine nucleotide releasing factor gene (RASGRF1) and the zinc finger, imprinted 2 gene (ZIM2). Despite the relatively small sample size used in these analyses, the observed associations with production traits are supported by the purported biological function of the RASGRF1 and ZIM2 gene products. These results support the hypothesis that imprinted genes contribute significantly to important complex production traits in cattle. Furthermore, these SNPs may be usefully incorporated into future marker-assisted and genomic selection breeding schemes.

Overcoming challenges of cancer treatment programmes in developing countries: a sustainable breast cancer initiative in Ethiopia.

The incidence of breast cancer is rising in many developing countries. Here we describe a programme to improve the support infrastructure for the management of patients with breast cancer in Addis Ababa, Ethiopia. Tamoxifen, a cheap, oral, yet effective, anti-cancer agent was made available freely to encourage staff and patients to follow well-defined, but achievable, protocols of care. Mammography, improved histopathological review, tissue hormone receptor assays, agreed treatment algorithms with a cycle of continuous audit of over 250 patients and cross-departmental patient management groups led to a considerable improvement in the management of breast cancer patients in a single institution. Aspects of this programme are now being extended to other regional hospitals in Ethiopia. Fairly limited investments in programmes for cancer can stimulate considerable improvements in the overall approach to malignant disease by encouraging a positive approach, even in very low resource environments.

Thymic expression of mutated B16:A preproinsulin messenger RNA does not reverse acceleration of NOD diabetes associated with insulin 2 (thymic expressed insulin) knockout.

We detected insulin2 mRNA but not insulin1 in thymus using real-time PCR analysis. Transgenic expression of a mutated insulin message (alanine rather than tyrosine at insulin B chain amino acid 16) was variably induced in thymus of four transgenic founder strains. The transgenic message levels were as high or higher than native insulin2 message. Lack of the insulin2 gene resulted in the enhancement of anti-insulin autoantibodies (regular NOD vs insulin2-knockout NOD, P<0.001) and in the presence of the B16:A insulin transgenes, levels of insulin autoantibodies remained elevated (regular NOD vs insulin2-knockout NOD with B16:A insulin, P<0.01). Diabetes acceleration by the knockout of the insulin2 gene was not influenced by the presence of the B16:A insulin transgenes. These data suggest that the B16:A insulin does not compensate for lack of native insulin expression in thymus. If lack of thymic insulin message of the insulin2 knockout is the cause of diabetes acceleration, this suggests that native insulin B:9-23 sequences may be crucial in thymus for insulin mediated immunomodulation. Further experiments varying native insulin message expression in thymus is necessary for direct comparison, but the current study provides additional evidence of the potential important role of a specific insulin B chain epitope.