Decreased expression of cell adhesion genes in cancer stem-like cells isolated from primary oral squamous cell carcinomas.

The goal of this study was to isolate cancer stem-like cells marked by high expression of CD44, a putative cancer stem cell marker, from primary oral squamous cell carcinomas and identify distinctive gene expression patterns in these cells. From 1 October 2013 to 4 September 2015, 76 stage III-IV primary oral squamous cell carcinoma of the gingivobuccal sulcus were resected. In all, 13 tumours were analysed by immunohistochemistry to visualise CD44-expressing cells. Expression of CD44 within The Cancer Genome Atlas-Head and Neck Squamous Cell Carcinoma RNA-sequencing data was also assessed. Seventy resected tumours were dissociated into single cells and stained with antibodies to CD44 as well as CD45 and CD31 (together referred as Lineage/Lin). From 45 of these, CD44+Lin- and CD44-Lin- subpopulations were successfully isolated using fluorescence-activated cell sorting, and good-quality RNA was obtained from 14 such sorted pairs. Libraries from five pairs were sequenced and the results analysed using bioinformatics tools. Reverse transcription quantitative polymerase chain reaction was performed to experimentally validate the differential expression of selected candidate genes identified from the transcriptome sequencing in the same 5 and an additional 9 tumours. CD44 was expressed on the surface of poorly differentiated tumour cells, and within the The Cancer Genome Atlas-Head and Neck Squamous Cell Carcinoma samples, its messenger RNA levels were higher in tumours compared to normal. Transcriptomics revealed that 102 genes were upregulated and 85 genes were downregulated in CD44+Lin- compared to CD44-Lin- cells in at least 3 of the 5 tumours sequenced. The upregulated genes included those involved in immune regulation, while the downregulated genes were enriched for genes involved in cell adhesion. Decreased expression of PCDH18, MGP, SPARCL1 and KRTDAP was confirmed by reverse transcription quantitative polymerase chain reaction. Lower expression of the cell-cell adhesion molecule PCDH18 correlated with poorer overall survival in the The Cancer Genome Atlas-Head and Neck Squamous Cell Carcinoma data highlighting it as a potential negative prognostic factor in this cancer.

Post-thyroidectomy suture granuloma: A diagnosis on fine needle aspiration cytology.

Suture granulomas result from the use of nonabsorbable suture and is an infrequent complication following thyroidectomy. When they occur, suture granulomas may mimic both benign and malignant diagnoses, posing a potential diagnostic challenge. Ultrasound-guided fine needle aspiration cytology is an accurate diagnostic modality for the workup of nodules within the thyroidectomy bed. We herein present 2 cases of post-thyroidectomy suture granulomas, presenting as a painless, palpable mass in the surgical bed and occurring as a late complication of thyroidectomy that were diagnosed on fine needle aspiration cytology.

Temporal and spatial expression of gonadotropin releasing hormone (GnRH) in the brain of developing zebrafish (Danio rerio).

Gonadotropin releasing hormone (GnRH) is a neuroendocrine decapeptide found in all vertebrate animals. The best understood function of GnRH is its endocrine function as a releasing hormone acting on the pituitary. But GnRH also functions as a neuromodulator within the nervous system. In a given species, GnRH occurs in a variety of forms that fall into three general categories based on pattern of protein/gene expression in the brain and amino acid sequence. The salmon GnRH (sGnRH), is found in the terminal nerve, where it has a neuromodulatory function. The sGnRH form is also the hypothalamic form in many fishes, although some fishes have a species specific form in the hypothalamus. Finally, chicken-GnRH-II (cGnRH-II), the most highly conserved form, is found in the midbrain where its function remains unclear. Here we have cloned the sGnRH and cGnRH-II cDNAs from zebrafish. By conducting stage specific in situ hybridization in developing zebrafish embryos, we provide a description of the spatial and temporal expression patterns of these genes. The location of sGnRH and cGnRH-II expressing cells is in agreement with previous reports of GnRH in the brains of adult fishes. Our results provide the first developmental description of GnRH gene expression where cGnRH-II and sGnRH are initially expressed at the onset of the first day after fertilization. Unlike what has been reported in many adult fishes, we did not find sGnRH expressed in the hypothalamic population during development.

De Novo transcriptome assembly (NGS) of Curcuma longa L. rhizome reveals novel transcripts related to anticancer and antimalarial terpenoids.

Herbal remedies are increasingly being recognised in recent years as alternative medicine for a number of diseases including cancer. Curcuma longa L., commonly known as turmeric is used as a culinary spice in India and in many Asian countries has been attributed to lower incidences of gastrointestinal cancers. Curcumin, a secondary metabolite isolated from the rhizomes of this plant has been shown to have significant anticancer properties, in addition to antimalarial and antioxidant effects. We sequenced the transcriptome of the rhizome of the 3 varieties of Curcuma longa L. using Illumina reversible dye terminator sequencing followed by de novo transcriptome assembly. Multiple databases were used to obtain a comprehensive annotation and the transcripts were functionally classified using GO, KOG and PlantCyc. Special emphasis was given for annotating the secondary metabolite pathways and terpenoid biosynthesis pathways. We report for the first time, the presence of transcripts related to biosynthetic pathways of several anti-cancer compounds like taxol, curcumin, and vinblastine in addition to anti-malarial compounds like artemisinin and acridone alkaloids, emphasizing turmeric's importance as a highly potent phytochemical. Our data not only provides molecular signatures for several terpenoids but also a comprehensive molecular resource for facilitating deeper insights into the transcriptome of C. longa.