Removal of nutrients and heavy metals from domestic and industry using botryococcus sp.

Microphytes or microalgae are the most basic food source of many types of organisms on earth and blooms during the presence of dissolved inorganic phosphorus. Wastewater is a body of water that is dangerous to organic life forms when consumed or used. It contains many pollutants that can cause health problems and also affect the ecosystem of an environment. This study aims to improve the water quality of wastewaters using phycoremediation process. The objectives of this study are to determine the growth of Botryococcus sp. in different types of wastewater in terms of resistance and survival of Botryococcus sp. in phycoremediation performance, to measure the environmental factor effecting the growth of Botryococcus sp. of phycoremediation process, to optimize the physiochemical and heavy metal removal in different types of wastewaters and to evaluate the effectiveness of Botryococcus sp. to remove the pollutants in wastewaters. Phycoremediation or bioremediation process is using macroalgae or microalgae for removing pollutants, nutrients, xenobiotics and heavy metals from wastewater. This research was done by collecting microalgae sample, isolating and culturing the required Botryococcus sp. Growth optimization and followed by phycoremediation process is done to remove unwanted elements from wastewaters. The optimum growth rate of algae is achieved when salinity is at 0M, temperature at 330C, photoperiod at 12:12 and light intensity of 18000 Lux. Result shows that the highest nitrate removal percentage occurs in semiconductor (100%), followed by palm oil mill effluent (97.29%), textile wastewater (98.04%) and domestic wastewater (85.43%). Total Phosphorus removal indicates the highest percentage for domestic wastewater (100%), palm oil effluent (99.2%), textile wastewater (98.44%) and semiconductor (50.39%). From this research, it is found that the best overall removal of physiochemical and heavy metal content occurs in palm oil mill effluent followed by domestic wastewater, semiconductor wastewater and textile wastewater

In Vitro Growth of Human Keratinocytes and Oral Cancer Cells into Microtissues: An Aerosol-Based Microencapsulation Technique

Cells encapsulation is a micro-technology widely applied in cell and tissue research, tissue transplantation, and regenerative medicine. In this paper, we proposed a growth of microtissue model for the human keratinocytes (HaCaT) cell line and an oral squamous cell carcinoma (OSCC) cell line (ORL-48) based on a simple aerosol microencapsulation technique. At an extrusion rate of 20 �L/min and air flow rate of 0.3 L/min programmed in the aerosol system, HaCaT and ORL-48 cells in alginate microcapsules were encapsulated in microcapsules with a diameter ranging from 200 to 300 �m. Both cell lines were successfully grown into microtissues in the microcapsules of alginate within 16 days of culture. The microtissues were characterized by using a live/dead cell viability assay, field emission-scanning electron microscopy (FE-SEM), fluorescence staining, and cell re-plating experiments. The microtissues of both cell types were viable after being extracted from the alginate membrane using alginate lyase. However, the microtissues of HaCaT and ORL-48 demonstrated differences in both nucleus size and morphology. The microtissues with re-associated cells in spheroids are potentially useful as a cell model for pharmacological studies

In vitro growth of human keratinocytes and oral cancer cells into microtissues: an aerosol-based microencapsulation technique

Cells encapsulation is a micro-technology widely applied in cell and tissue research, tissue transplantation, and regenerative medicine. In this paper, we proposed a growth of microtissue model for the human keratinocytes (HaCaT) cell line and an oral squamous cell carcinoma (OSCC) cell line (ORL-48) based on a simple aerosol microencapsulation technique. At an extrusion rate of 20 µL/min and air flow rate of 0.3 L/min programmed in the aerosol system, HaCaT and ORL-48 cells in alginate microcapsules were encapsulated in microcapsules with a diameter ranging from 200 to 300 µm. Both cell lines were successfully grown into microtissues in the microcapsules of alginate within 16 days of culture. The microtissues were characterized by using a live/dead cell viability assay, field emission-scanning electron microscopy (FE-SEM), fluorescence staining, and cell re-plating experiments. The microtissues of both cell types were viable after being extracted from the alginate membrane using alginate lyase. However, the microtissues of HaCaT and ORL-48 demonstrated differences in both nucleus size and morphology. The microtissues with re-associated cells in spheroids are potentially useful as a cell model for pharmacological studies

Zebrafish phenotypic screen identifies novel Notch antagonists

Zebrafish represents a powerful in vivo model for phenotype-based drug discovery to identify clinically relevant small molecules. By utilizing this model, we evaluated natural product derived compounds that could potentially modulate Notch signaling that is important in both zebrafish embryogenesis and pathogenic in human cancers. A total of 234 compounds were screened using zebrafish embryos and 3 were identified to be conferring phenotypic alterations similar to embryos treated with known Notch inhibitors. Subsequent secondary screens using HEK293T cells overexpressing truncated Notch1 (HEK293TΔE) identified 2 compounds, EDD3 and 3H4MB, to be potential Notch antagonists. Both compounds reduced protein expression of NOTCH1, Notch intracellular domain (NICD) and hairy and enhancer of split-1 (HES1) in HEK293TΔE and downregulated Notch target genes. Importantly, EDD3 treatment of human oral cancer cell lines demonstrated reduction of Notch target proteins and genes. EDD3 also inhibited proliferation and induced G0/G1 cell cycle arrest of ORL-150 cells through inducing p27KIP1. Our data demonstrates the utility of the zebrafish phenotypic screen and identifying EDD3 as a promising Notch antagonist for further development as a novel therapeutic agent

Transcriptional analysis highlights three distinct immune profiles of high-risk oral epithelial dysplasia

Oral potentially malignant disorders (OPMD) are precursors of oral squamous cell carcinoma (OSCC), and the presence of oral epithelial dysplasia (OED) in OPMD confers an increased risk of malignant transformation. Emerging evidence has indicated a role for the immune system in OPMD disease progression; however, the underlying immune mechanisms remain elusive. In this study, we used immune signatures established from cancer to delineate the immune profiles of moderate and severe OED, which are considered high-risk OPMD. We demonstrated that moderate and severe OEDs exhibit high lymphocyte infiltration and upregulation of genes involved in both immune surveillance (major histocompatibility complex-I, T cells, B cells and cytolytic activity) and immune suppression (immune checkpoints, T regulatory cells, and tumor-associated macrophages). Notably, we identified three distinct subtypes of moderate and severe OED: immune cytotoxic, non-cytotoxic and non-immune reactive. Active immune surveillance is present in the immune cytotoxic subtype, whereas the non-cytotoxic subtype lacks CD8 immune cytotoxic response. The non-immune reactive subtype showed upregulation of genes involved in the stromal microenvironment and cell cycle. The lack of T cell infiltration and activation in the non-immune reactive subtype is due to the dysregulation of CTNNB1, PTEN and JAK2. This work suggests that moderate and severe OED that harbor the non-cytotoxic or non-immune reactive subtype are likely to progress to cancer. Overall, we showed that distinct immune responses are present in high-risk OPMD, and revealed targetable pathways that could lead to potential new approaches for non-surgical management of OED

Overexpression of MMP13

Matrix metalloproteinase 13 (MMP13) plays a central role in the MMP activation cascade that enables degradation of the extracellular matrix and basement membranes, and it is identified as a potential driver in oral carcinogenesis. Therefore, this study aims to determine the copy number, mRNA, and protein expression of MMP13 in oral squamous cell carcinoma (OSCC) and to associate these expressions with clinicopathological parameters. Copy number, mRNA, and protein expression analysis of MMP13 were determined using real-time quantitative PCR and immunohistochemistry methods in OSCC samples. The correlations between MMP13 expressions and clinicopathological parameters were evaluated, and the significance of MMP13 as a prognostic factor was determined. Despite discrepancies between gene amplification and mRNA and protein overexpression rates, OSCC cases showed high amplification of MMP13 and overexpression of MMP13 at both mRNA and protein levels. High level of MMP13 protein expression showed a significant correlation with lymph node metastasis (P=0.011) and tumor staging (P=0.002). Multivariate Cox regression model analysis revealed that high level of mRNA and protein expression of MMP13 were significantly associated with poor prognosis (P<0.050). Taken together, these observations indicate that the MMP13 protein overexpression could be considered as a prognostic marker of OSCC

Oral mucosal lesion detection accuracy post lectures and tests in clinical dental students

The oral medicine field of dentistry comprises learning to diagnose oral soft tissue disease and is taught from the third year of dental school. Despite long-term learning on oral medicine, there are no data on the clinical accuracy of oral mucosal lesion (OML) diagnosis by clinical dental students (CDS). Purpose: to evaluate the effectiveness of oral medicine lectures prior to community service in Tanjung Pandan, Indonesia. Methods: An observational study was done by 60 CDS, divided into 3 groups, who were tested and/or given prior lectures. OML detection was performed by CDS and re-confirmed by an oral medicine specialist. The analyses were done by t-test, ANOVA, and Cohen's Kappa. The results: Out of 615 patients, only 243 patients had OML. There was a significant difference in test scores found between groups with or without prior lectures(P=.026; P=.015). The accuracy and inter-agreement of OML detection was good with substantial agreement (AUC=.825; K=.629); however, there was fair agreement on normal variant oral lesions (AUC=.68; k=.322), and all groups failed to detect and diagnose OPMD lesions (AUC=.501; k=.003). There was a positive correlation between test #2 and AUC OML (R=.845), and with every increase in test score, the accuracy is expected to be .033 higher. Dental student OML knowledge should be upgraded by frequent oral lesion case practice. Further educational strategy is needed to develop dental student's knowledge and skill, so that they can integrate their learning into their practice

Reply to “Letter to the Editor”

Dear Editor, We thank Rentoft and colleagues for commenting on our paper “Transcriptional profiling of oral squamous cell carcinoma using formalin-fixed paraffin-embedded samples”, and agreeing that formalin-fixed paraffin embedded (FFPE) tissue specimens can be used for gene expression studies using microarrays, to identify genes that are significantly involved in oral carcinogenesis. The similarities and high concordance between the study by Rentoft et al. [1] and ours [2] is comforting and clearly supports the use of FFPE tissues in such experiments. More importantly, these studies act as independent validation for one another and strongly suggests that genes that were found to be up- or down-regulated in oral squamous cell carcinoma (OSCC) do indeed play a role in these cancers and therefore warrant further investigation to determine their utility as biomarkers and therapeutic targets for OSCC. We wanted to point out however, that the similarities between these 2 studies are not completely unexpected despite previous reports highlighting that the concordance between microarray studies are hard to achieve [3]. The main disparities between datasets from microarray experiments have been attributed to the use of different microarray platforms and the heterogeneity of the tissue specimens that were used [4], [5]. Indeed, many microarray studies reported for the head and neck, used tissues from several distinct areas which have been reported to be genetically heterogeneous, and associated with different aetiologies [6], [7]. Given that both our studies used the DASL assay and tissues from the oral cavity (albeit from different sites-explained further below) the consistency of the genes that were identified should not come as a complete surprise. However, it is still intriguing that the similarities between these two studies were so close despite previous reports indicating that there are distinct differences between oral cancers associated with different aetiology [8], [9], and reports describing the distinct genetic differences between subsites of the oral cavity [10], [11], [12]. Our previous study describing the differences in gene-expression patterns between oral cancers associated with betel quid chewing and smoking demonstrated that despite the differences seen, genetic changes common to all the cancers were also observed suggesting that there are core events and pathways that are important regardless of the aetiology or site of the cancer [8]. Consistently, upon close examination of the genes that are most differentially expressed between our study and that of Rentoft et al. [1] 5 of the top 10 over-lapping genes most differentially expressed were matrix metalloproteinases, whilst other genes include IL-8, CXCL-9 and BCL2A1, genes that were consistently up-regulated in many other microarray studies of the head and neck independent of the subsites of cancer [8], [12], [13], [14]. An unprecedented large overlap of genes between our study [2] and that of Rentoft and colleagues [1] may also be due to the smaller number of genes on the arrays that were used in these studies (502 genes), and that these genes were pre-selected based on their involvement in cancer development, whereas the majority of previous studies used platforms consisting of larger arrays and hence could capture much more of the heterogeneity typically observed in cancers. More recently, Illumina launched its whole genome DASL assay and it would be interesting to see if the similarities between buccal and tongue cancers still prevails. In conclusion, increasing number studies including the 2 compared here, strongly supports the use of FFPE tissues for gene expression studies using microarrays. The development of new technologies and statistical methods that addresses the challenges associated with using FFPE tissues, combined with the clinical information available with these specimens, will indeed facilitate the identification and discovery of clinically relevant gene signatures and biomarkers and therapeutic targets to improve the management of cancers in general