Methylation study of tumor suppressor genes in human aberrant crypt foci, colorectal carcinomas, and normal colon.

BACKGROUND: Aberrant crypt foci (ACF) are the earliest preneoplastic lesions in human colon, identifiable on chromoendoscopic screening. Our objective was to evaluate the %methylation of APC, CDKN2A, MLH1, RASSF1, MGMT, and WIF1 tumor suppressor genes (TSG) in ACF, corresponding colorectal carcinomas (CRC), and normal colonic mucosal controls. METHODS: In this study, macroscopically normal-appearing mucosal flaps were sampled 5-10 cm away from the tumor mass from 302 fresh colectomy specimens to identify ACF-like lesions. Thirty-five cases with multiple ACFs were selected (n 35) as the main study group, with corresponding sections from CRC (n 35) as disease controls, and mucosal tissue blocks from 20 colectomy specimens (normal controls), operated for non-neoplastic pathologies. Genomic DNA was extracted, and methylation-specific polymerase chain reaction (PCR) was performed on a customized methylation array model. %Methylation data were compared among the groups and with clinicopathological parameters. Selected target mRNA and protein expression studies were performed. RESULTS: %Methylation of TSGs in ACF was intermediate between normal colon and CRC, although a statistically significant difference was observed only for the WIF1 gene (P < 0.01). Also, there was increased nuclear ß-catenin expression and upregulation of CD44-positive cancer-stem cells in ACF and CRCs than in controls. Right-sided ACFs and dysplastic ACFs had a higher %methylation of CDKN2A (P < 0.01), whereas hyperplastic ACFs had a higher %methylation of RASSF1 (P 0.04). The topographic characteristics of ACFs did not correlate with TSG %methylation. CONCLUSIONS: Early epigenetic methylation of WIF1 gene is one of the mechanisms for ACF development in human colon.

Using Human Fetal Neural Stem Cells to Elucidate the Role of the JAK-STAT Cell Signaling Pathway in Oligodendrocyte Differentiation In Vitro.

Oligodendrocytes (OL) are the myelinating cells of the central nervous system that mediate nerve conduction. Loss of oligodendrocytes results in demyelination, triggering neurological deficits. Developing a better understanding of the cell signaling pathways influencing OL development may aid in the development of therapeutic strategies. The primary focus of this study was to investigate and elucidate the cell signaling pathways implicated in the developmental maturation of oligodendrocytes using human fetal neural stem cells (hFNSCs)-derived primary OL and MO3.13 cell line. Successful differentiation into OL was established by examining morphological changes, increased expression of mature OL markers MBP, MOG and decreased expression of pre-OL markers CSPG4 and O4. Analyzing transcriptional datasets (using RNA sequencing) in pre-OL and mature OL derived from hFNSCs revealed the novel and critical involvement of the JAK-STAT cell signaling pathway in terminal OL maturation. The finding was validated in MO3.13 cell line whose differentiation was accompanied by upregulation of IL-6 and the transcription factor STAT3. Increased phosphorylated STAT3 (pY705) levels were demonstrated by western blotting in hFNSCs-derived primary OL as well as terminal maturation in MO3.13 cells, thus validating the involvement of the JAK-STAT pathway in OL maturation. Pharmacological suppression of STAT3 phosphorylation (confirmed by western blotting) was able to prevent the increase of MBP-positive cells as demonstrated by flow cytometry. These novel findings highlight the involvement of the JAK-STAT pathway in OL maturation and raise the possibility of using this as a therapeutic strategy in demyelinating diseases.

Harnessing the potential of white rot fungi and ligninolytic enzymes for efficient textile dye degradation: A comprehensive review.

The contamination of wastewater with textile dyes has emerged as a pressing environmental concern due to its persistent nature and harmful effects on ecosystems. Conventional dye treatment methods have proven inadequate in effectively breaking down complex dye molecules. However, a promising alternative for textile dye degradation lies in the utilization of white rot fungi, renowned for their remarkable lignin-degrading capabilities. This review provides a comprehensive analysis of the potential of white rot fungi in degrading textile dyes, with a particular focus on their ligninolytic enzymes, specifically examining the roles of lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase in the degradation of lignin and their applications in textile dye degradation. The primary objective of this paper is to elucidate the enzymatic mechanisms involved in dye degradation, with a spotlight on recent research advancements in this field. Additionally, the review explores factors influencing enzyme production, including culture conditions and genetic engineering approaches. The challenges associated with implementing white rot fungi and their ligninolytic enzymes in textile dye degradation processes are also thoroughly examined. Textile dye contamination poses a significant environmental threat due to its resistance to conventional treatment methods. White rot fungi, known for their ligninolytic capabilities, offer an innovative approach to address this issue. The review delves into the intricate mechanisms through which white rot fungi and their enzymes, including LiP, MnP, and laccase, break down complex dye molecules. These enzymes play a pivotal role in lignin degradation, a process that can be adapted for textile dye removal. The review also emphasizes recent developments in this field, shedding light on the latest findings and innovations. It discusses how culture conditions and genetic engineering techniques can influence the production of these crucial enzymes, potentially enhancing their efficiency in textile dye degradation. This highlights the potential for tailored enzyme production to address specific dye contaminants effectively. The paper also confronts the challenges associated with integrating white rot fungi and their ligninolytic enzymes into practical textile dye degradation processes. These challenges encompass issues like scalability, cost-effectiveness, and regulatory hurdles. By acknowledging these obstacles, the review aims to pave the way for practical and sustainable applications of white rot fungi in wastewater treatment. In conclusion, this comprehensive review offers valuable insights into how white rot fungi and their ligninolytic enzymes can provide a sustainable solution to the urgent problem of textile dye-contaminated wastewater. It underscores the enzymatic mechanisms at play, recent research breakthroughs, and the potential of genetic engineering to optimize enzyme production. By addressing the challenges of implementation, this review contributes to the ongoing efforts to mitigate the environmental impact of textile dye pollution. PRACTITIONER POINTS: Ligninolytic enzymes from white rot fungi, like LiP, MnP, and laccase, are crucial for degrading textile dyes. Different dyes and enzymatic mechanisms is vital for effective wastewater treatment. Combine white rot fungi-based strategies with mediator systems, co-culturing, or sequential treatment approaches to enhance overall degradation efficiency. Emphasize the broader environmental impact of textile dye pollution and position white rot fungi as a promising avenue for contributing to mitigation efforts. This aligns with the overarching goal of sustainable wastewater treatment practices and environmental conservation. Consider scalability, cost-effectiveness, and regulatory compliance to pave the way for sustainable applications that can effectively mitigate the environmental impact of textile dye pollution.

Comparative evaluation of anti-biofilm and anti- adherence potential of plant extracts against Streptococcus mutans: A therapeutic approach for oral health.

Dental caries predominantly attributed to the cariogenic nature of Streptococcus mutans, continue to pose a substantial global challenge to oral health. In response to this challenge, this study aimed to evaluate the effectiveness of leaf extracts (LEs) and essential oils (EOs) derived from different medicinal plants in inhibiting the growth of Streptococcus mutans biofilm. In vitro and in silico approaches were employed to identify active compounds and assess their inhibitory effects on S. mutans. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were measured to determine the anti-biofilm and anti-adherence activity against S. mutans. Biofilm viability (CFU/mL) and extracellular polymeric substance (EPS) concentration were quantified. GC-MS analysis was utilized to identify active compounds in the most effective plant extracts exhibiting anti-S. mutans activity. A high-throughput screening focused on the interaction between these compounds and the target enzyme SortaseA (SrtA) using molecular docking was performed. Results indicated that Cymbopogon citratus displayed the highest efficacy in reducing S. mutans biofilm formation and adhesion activity, achieving 90 % inhibition at an MIC value of 12 µg/mL. Among the 12 bioactive compounds identified, trans-Carvyl acetate exhibited the lowest binding energy with SrtA (-6.0 Kcal/mole). Trans-Carvyl acetate also displayed favorable pharmacokinetic properties. This study provides novel insights into the anti-S. mutans properties of C. citratus and suggests its potential as a therapeutic approach for oral health. Further research is needed to explore the combined effect of plant extracts for enhanced protection against dental caries.

Deciphering the mechanism of anti-quorum sensing post-biotic mediators against Streptococcus mutans.

OBJECTIVE: Glucosyltransferases (Gtfs) and quorum sensing (QS) mediated transduction genes play critical roles in the pathogenesis of Streptococcus mutan-mediated dental caries. Therefore, targeting gtfs and QS-mediated virulence genes have therefore emerged as an intriguing goal for efficient therapeutic approaches that block cariogenic biofilms. METHODS: Post-biotic mediators (PMs) obtained from our previously isolated and characterized beneficial bacteria Enterobacter colacae PS-74 was assessed for its antibiofilm potential against S. mutans. According to the transcriptome method, qRT-PCR analysis was performed against virulence genes. For microscopic visualization, SEM and CLSM analyses were used to confirm the inhibitory effects of PMs. RESULTS: PMs dramatically reduced the expression of QS signal transduction, glucan metabolism, and biofilm-regulated genes such gtfB, gtfC, ComDE, VicR, brpA in S. mutans, which validates the outcomes of in vitro result. Their unique metabolites may help to control biofilm formation by eluding antimicrobial resistance. CONCLUSION: Considering the above findings, PMs may deem to be an innovative, alluring, and secure method for preventing dental caries due to their biological activity. Our study unravels the inhibitory effect of PMs, which will contribute to instruct drug design strategies for effective inhibition of S. mutans biofilms.

Phenotypic Profiling of Immune Cells and Their Mediators in Chronic Obstructive Pulmonary Disease.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is an inflammatory disorder and has been proposed to have an imbalance between pro-inflammatory and anti-inflammatory factors. METHODS: This study was conducted on 41 participants {18 COPD patients (smokers, COPD S (n = 9); reformed smokers, COPD RS (n = 9)) and 23 controls (non-smokers, CNS (n = 14); smokers, CS (n = 9))}. Flow cytometry was used to identify circulatory immune cells and correlated with serum cytokines. RESULTS: On comparison, significantly lower frequency of CD3+ T cells were observed in COPD S as compared to CNS (p < 0.01) and CS (p < 0.01); CD4+ T cells were lower in COPD S (p < 0.05), COPD RS (p < 0.05) and CNS (p < 0.01) as compared to CS. CD8+ T cells were elevated in COPD S as compared to CS (p < 0.05). Lower frequency of cDCs were observed in COPD S as compared to CS (p < 0.05) and COPD RS as compared to CNS (p < 0.01) and CS (p < 0.01). Lower frequency of pDCs were observed in COPD RS as compared to COPD S (p < 0.05), CNS (p < 0.05) and CS (p < 0.01). Lower frequency of Tregs was observed in COPD S as compared to CNS (p < 0.05) and CS (p < 0.05). CONCLUSIONS: Characteristic changes observed indicate a significant impact of immune cells in the progression of the disease.

Fortification of orange juice with microencapsulated Kocuria flava Y4 towards a novel functional beverage: Biological and quality aspects.

To commercialize functional foods, probiotics must exhibit high resistance and acceptable stability under various unfavorable conditions to maintain the quality of fruit juices. This study will provide an insight into fortification of orange juice with a plant probiotic Kocuria flava Y4 by microencapsulation. Therefore, this study investigated the colony release, physicochemical and phytochemical parameters, and antioxidant activity of the orange juice exposed to microencapsulated probiotics and the one without probiotics (control). Evaluation of orange juice on the growth of probiotic bacteria showed that the fortification with alginate and psyllium micro-particles showed highest encapsulation efficiency (99.01%) and acceptable viability of probiotic cells (8.12 ± 0.077 CFU/mL) during five weeks storage at 4 °C. The morphology and functional properties of beads was studied by SEM, Zeta-potential and FTIR analysis. The sucrose and organic acids concentrations decreased significantly during fortification period (0-72 h) except ascorbic acid. Furthermore, glucose, pH, acidity, TSS were maintained. The results affirm the suitability and feasibility of developing a plant probiotic beverage using orange juice by encapsulation method.

Deciphering the role of fungus in degradation of polypropylene from hospital waste.

Health and environmental consequences are unavoidable when it comes to management of hospital waste (HW) disposables. In order to eradicate the HW, this study isolated a novel fungus SPF21 from a hospital dumping yard to degrade Polypropylene (PP). We measured the attributes of PP inoculated with fungus using mass loss, Fourier trans-form infrared (FTIR), contact angle (CA), and scanning electron microscopy (SEM). The weight of PP exposed to SPF21 was reduced by 25% in 90 days. The SEM images reveal that there are pores all over the sample surface; they alsocaused voids during the biodegradation of PP. FTIR analysis indicates that the spectra of treated mask pieces show the absence of peak at 1746 cm-1 and the appearance of a new peak at 1643 cm-1 . A period of 90-day exposure to the fungal isolate SPF21 reduced the CA of PP by 44.8% when compared to the nonexposed PP samples, suggesting that the surface of PP turned more hydrophilic after exposure. Moreover, our study on PP degradation by the fungus Ascotricha sinuosa SPF21 appears to be promising from the perspective of environmental, health, and economic hazards. Our results indicate that biodegradation greatly facilitates fungus deposition and changes PP film morphology and hydrophilicity.

Mycoremediation of anthraquinone dyes from textile industries: a mini-review.

The discharge of wastewater from textile industries into aquatic bodies has severe health and environmental impacts. Textile industries generate huge amounts of effluents containing hazardous toxic dyes. Anthraquinone (AQ) dyes containing AQ chromophore groups are the second most important class of nondegradable textile dyes, preceded by azo dyes. Despite their prevalence, biodegradation of AQ dyes has not yet been completely understood because of their complex and stable structures. Currently, microbiological approaches to treating dyeing wastewater are considered economical and feasible, and reports regarding fungal degradation of AQ dyes are increasing. Structures and classification of AQ dyes were summarized in this study along with degradative fungi, and their enzyme systems with influencing factors and possible mechanisms of AQ mycoremediation were explored. Furthermore, the existing problems and present research progress were discussed. Finally, the key points with future research directions were presented.

Hypoxia Induces Early Neurogenesis in Human Fetal Neural Stem Cells by Activating the WNT Pathway.

Fetal neural stem cells (FNSCs) present in the human fetal brain differentiate into cells of neuronal and glial lineages. The developing fetus is exposed to lower oxygen concentrations than adults, and this physiological hypoxia may influence the growth and differentiation of the FNSCs. This study aimed to evaluate the effect of hypoxia on the differentiation potential of human FNSCs isolated from the subventricular zone of aborted fetal brains (n = 5). FNSCs were isolated, expanded, and characterized by Nestin and Sox2 expression using immunocytochemistry and flow cytometry, respectively. These FNSCs were exposed to 20% oxygen (normoxia) and 0.2% oxygen (hypoxia) concentrations for 48 h, and hypoxia exposure (n = 5) was validated. Whole transcriptome analyses (Genespring GX13) of FNSCs exposed to hypoxia (Agilent 4 × 44 K human array slides) highlighted that genes associated with neurogenesis were enriched upon exposure to hypoxia. The pathway analysis of these enriched genes (using Metacore) showed the involvement of the WNT signaling pathway. Microarray analyses were validated using neuronal and glial lineage commitment markers, namely, NEUROG1, NEUROG2, ASCL1, DCX, GFAP, OLIG2, and NKX2.2, using qPCR (n = 9). DCX, ASCL1, NGN1, and GFAP protein expression was analyzed by Western blotting (n = 3). This demonstrated upregulation of the neuronal commitment markers upon hypoxia exposure, while no change was observed in astrocytic and oligodendrocyte lineage commitment markers. Increased expression of downstream targets of the WNT signaling pathway, TCF4 and ID2, by qPCR (n = 9) and increased protein expression of CTNNB1 (ß-catenin) and ID2 by Western blot (n = 3) indicated its involvement in mediating neuronal differentiation upon exposure to hypoxia.

Probiotic Characterization of Indigenous Kocuria flava Y4 Strain Isolated from Dioscorea villosa Leaves.

This aim of the study was to isolate and screen potential probiotics from Dioscorea villosa leaves. The potential isolate Y4 was obtained from the Dioscorea villosa leaves, and its ability to grow in a medium containing high NaCl concentrations (2-10%) indicated its negative hemolytic activity. Furthermore, Y4 demonstrated inhibitory activity against human pathogens, such as Klebsiella pneumonia, Staphylococcus aureus, Citrobacter koseri, and Vibrio cholerae, as well as towards a plant pathogen isolate OR-2 (obtained from Citrus sinensis). Some biologically important functional groups of Y4 metabolites, such as sulfoxide; aliphatic ether; 1, 2, 3-trisubstituted, tertiary alcohol: vinyl ether; aromatic amine; carboxylic acid; nitro compound; alkene mono-substituted; and alcohol, were identified through FTIR analysis. The 16S rRNA sequencing and subsequent phylogenetic tree analysis indicated that Y4 and OR-2 are the closest neighbors to Kocuria flava (GenBank accession no. MT773277) and Pantoea dispersa (GenBank accession no. MT766308), respectively. The potential isolate Y4 was found to exhibit adhesion, auto-aggregation, co-aggregation, and weak biofilm activity. It also exhibited a high level of antimicrobial activity and antibiotic susceptibility. The safety of K. flava Y4 isolate, which is proposed to be a probiotic, was evaluated through acute oral toxicity test and biogenic amine production test. Moreover, the preservation potential of isolate Y4 was assessed through application on fruits under different temperatures. Thus, our results confirmed that Kocuria flava Y4 is a prospective probiotic and could also be used for the preservation of fruits.

Case report: Characterization of a rare pathogenic variant associated with loss of COL3A1 expression in vascular Ehlers Danlos syndrome.

The vascular subtype of Ehlers Danlos Syndrome (vEDS) is a rare connective tissue disorder characterized by spontaneous arterial, bowel or organ rupture. The diagnosis of vEDS is established in a proband by identification of a heterozygous pathogenic variant in the alpha-1 gene of type III collagen (COL3A1) by molecular analysis. In this report, we present a case of vEDS with life threatening, spontaneous arterial dissections in association with an uncharacterized rare variant of COL3A1, exon19:c.1340G > A. Primary culture of patient skin fibroblasts followed by immunofluorescence revealed a complete absence of COL3A1 protein expression as well as altered morphology. Electron microscopy of the cultured fibroblasts showed abnormal vacuoles in the cytoplasm suggestive of a secretory defect. In this study, we have performed functional characterization of the COL3A1 exon19:c.1340G > A variant for the first time and this may now be classified as likely pathogenic in vEDS. *Both JM and LRL contributed equally in the manuscript and should both be considered as the first author.

Protocol for Developing a Femur Osteotomy Model in Wistar Albino Rats.

Fracture healing is a physiological process resulting in the regeneration of bone defects by the coordinated action of osteoblasts and osteoclasts. Osteoanabolic drugs have the potential to augment the repair of fractures but have constraints like high costs or undesirable side effects. The bone healing potential of a drug can initially be determined by in vitro studies, but in vivo studies are needed for the final proof of concept. Our objective was to develop a femur osteotomy rodent model that could help researchers understand the development of callus formation following fracture of the shaft of the femur and that could help establish whether a potential drug has bone healing properties. Adult male Wistar albino rats were used after Institutional Animal Ethics Committee clearance. The rodents were anesthetized, and under aseptic conditions, complete transverse fractures at the middle one-third of the shafts of the femurs were created using open osteotomy. The fractures were reduced and internally fixed using intramedullary K-wires, and secondary fracture healing was allowed to take place. After surgery, intraperitoneal analgesics and antibiotics were given for 5 days. Sequential weekly x-rays assessed callus formation. The rats were sacrificed based on radiologically pre-determined time points, and the development of the fracture callus was analyzed radiologically and using immunohistochemistry.

Glutamate Uptake Is Not Impaired by Hypoxia in a Culture Model of Human Fetal Neural Stem Cell-Derived Astrocytes.

Hypoxic ischemic injury to the fetal and neonatal brain is a leading cause of death and disability worldwide. Although animal and culture studies suggest that glutamate excitotoxicity is a primary contributor to neuronal death following hypoxia, the molecular mechanisms, and roles of various neural cells in the development of glutamate excitotoxicity in humans, is not fully understood. In this study, we developed a culture model of human fetal neural stem cell (FNSC)-derived astrocytes and examined their glutamate uptake in response to hypoxia. We isolated, established, and characterized cultures of FNSCs from aborted fetal brains and differentiated them into astrocytes, characterized by increased expression of the astrocyte markers glial fibrillary acidic protein (GFAP), excitatory amino acid transporter 1 (EAAT1) and EAAT2, and decreased expression of neural stem cell marker Nestin. Differentiated astrocytes were exposed to various oxygen concentrations mimicking normoxia (20% and 6%), moderate and severe hypoxia (2% and 0.2%, respectively). Interestingly, no change was observed in the expression of the glutamate transporter EAAT2 or glutamate uptake by astrocytes, even after exposure to severe hypoxia for 48 h. These results together suggest that human FNSC-derived astrocytes can maintain glutamate uptake after hypoxic injury and thus provide evidence for the possible neuroprotective role of astrocytes in hypoxic conditions.

Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4.

The exogenous lipolytic activities of Kocuria sp. have been recognized earlier but the genus further contains many more unexplored strains. In this study, the extracellular lipase activity of Kocuria flava Y4 (GenBank accession no. MT773277), isolated from Dioscorea villosa during our previous study, was regulated by different physicochemical parameters, such as pH, temperature, shaking speed, and incubation time. For efficient immobilization of the extracellular lipase, 4% sodium alginate, 50 mL of 25 nM CaCl2.2H2O solution, and 15 min. Hardening time of gel beads in calcium chloride was used. For the first time, K. flava Y4 lipase was purified using ammonium sulphate precipitation followed by dialysis and DEAE-Sepharose anion exchange chromatography with Sepharose-6B gel filtration chromatography, yielding ∼15-fold purified lipase with a final yield of 96 U/mL. The SDS-PAGE of purified lipase displayed a single strong band, indicating a monomeric protein of 45 kDa. At a temperature of 35°C and pH 8, the purified lipase showed maximum hydrolytic activity. Using p-nitrophenyl acetate (p-NPA) as the hydrolysis substrate, the values of K m and V max derived from the Lineweaver-Burk plot were 4.625 mM and 125 mol/min-1mg-1, respectively.

Mycoremediation and toxicity assessment of textile effluent pertaining to its possible correlation with COD.

Globally, textile industries are one of the major sectors releasing dye pollutants. This is the first report on the positive correlation between toxicity and chemical oxygen demand (COD) of textile effluent along with the proposed pathway for enzymatic degradation of acid orange 10 using Geotrichum candidum within a very short stretch of time (18 h). Removal efficiency of this mycoremedial approach after 18 h in terms of chemical oxygen demand, biological oxygen demand, total suspended solids, salinity, color and dye concentration in the treated effluent reached to 98.5%, 56.3%,73.2%, 64%, 89% and 87% respectively. Also there was a decrease in pH of the treated effluent. FTIR analysis of the treated effluent confirmed biodegradation. The LCMS analysis showed the degradation of acid orange 10, which was confirmed by the formation of two biodegradation products, 7-oxo-8-iminonapthalene-1,3-disulfonate and nitrosobenzene, which subsequently undergoes stepwise hydrogenation and dehydration to form aniline via phenyl hydroxyl amine as intermediate. The X-ray diffraction studies showed that heavy metal content in the treated effluent has reduced along with decrease in % crystallinity, indicating biodegradation. The connection between toxicity and COD was also inveterated using Pearson's correlation coefficient. Further the toxicological studies indicated the toxicity of raw textile effluent and relatively lower toxic nature of metabolites generated after biodegradation by G. candidum.

Degradation of dyes by fungi: an insight into mycoremediation.

Currently, globalization, urbanization and industrialization have led to several environmental issues. In many industries, particularly in textile industries, the extensive use of synthetic dyes has increased. Dye is an integral element used to impart color to textile materials. Wastes generated during the processing and treatment of the dye contain inorganic and organic compounds that are hazardous, thereby posing a serious threat to the ecosystem. It is therefore important to implement cost-efficient and successful measures against these emissions in order to preserve habitats and natural resources. In this context, biodegradation by fungi or mycoremediation of dyes using potential fungi is a fairly inexpensive and environmental friendly method for decomposing or mineralizing barely or less decaying dye compounds. Fungi play a crucial role in degrading and decolorizing organic dyes by enzymes and processes such as absorption, adsorption and aggregation of effluent colorants. The factors affecting the decolorization and biodegradation of dye compounds through fungal bioremediation, such as pH, temperature, dye concentration, agitation, effects of carbon and nitrogen sources, dye structure, enzymes, electron donor and redox mediators are discussed in this review. The review also includes a summary on the mechanism and kinetics of dye degradation as well as recent advances and future perspectives in mycoremediation of dyes.

Comparing Migratory and Mechanical Properties of Human Bone Marrow-Derived Mesenchymal Stem Cells with Colon Cancer Cells In Vitro.

BACKGROUND: Colon cancer cells can migrate and metastasize by undergoing epithelial-to-mesenchymal transition (EMT). Mesenchymal stem cells (MSCs) are non-cancerous, multipotent adult stem cells, which can also migrate. In this study, we wanted to compare the biological, physical, and functional properties of these migratory cells. MATERIALS AND METHODS: HT-29 and HCT-116, two human colon carcinoma cell lines, represent less aggressive and more aggressive cancer cells, respectively. MSCs were isolated from human bone marrow. After confirming the identity of all the cell types, they were evaluated for E-cadherin, ß1-integrin, Vimentin, ZEB-1, ß-catenin, and 18S rRNA using Q-PCR. MMP-2 and MMP-9 activity were evaluated using gelatin zymography. Functional tests like wound healing assay, migration assay, and invasion assay were also done. Biomechanical properties like cell stiffness and non-specific adhesion (between indenter probe and cell membrane) were evaluated through nanoindentation using atomic force microscopy (AFM). RESULTS: Expression of EMT and stem cell markers showed typical expression patterns for HT-29, HCT-116, and MSCs. Functional tests showed that MSCs migrated faster than malignant cells. MMP-2 and MMP-9 activity reinforced this behavior. Interestingly, the migration/invasion capacity of MSCs was comparable to aggressive HCT-116, and more than HT-29. MSCs also showed the maximum cell stiffness and non-specific cell-probe adhesions, followed by HCT116 and HT29 cells. CONCLUSIONS: Our findings indicate that the migratory properties of MSCs is comparable or even greater than that of cancer cells and despite their high migration potential, they also have the maximum stiffness.

Increased Mucin Expression in Oral Mucosal Epithelial Cells in vitro: A Potential New Role of Mycophenolate Mofetil.

OBJECTIVE: Autologous cultured explants of human oral mucosal epithelial cells (OMEC) are a potential therapeutic modality in patients of bilateral ocular surface disease (OSD) with incapacitating dry eye. Mycophenolate mofetil (MMF) has been found to upregulate the mucin production in conjunctival goblet cells in vitro. The aim of this study was to evaluate the effects of MMF on mucin expression in primary cultures of OMEC. METHODS: With informed consent, oral mucosal epithelial tissue samples were obtained from patients undergoing oral surgery for non-malignant conditions. OMEC were cultured on human amniotic membrane (HAM) scaffold for 2 weeks. Mucin expression was quantified using RT-PCR and qPCR before and after treating cultured OMEC with MMF. RESULTS: Morphological studies revealed a confluent sheet of proliferating, stratified oral mucosal epithelial cells. Mucin mRNAs were elucidated by RT-PCR. Compared to untreated controls, MUC1, MUC15 and MUC16 mRNAs and MUC1 protein expression were found to be upregulated in MMF treated primary cultures of OMEC, as assessed by qPCR and immunocytochemistry respectively. CONCLUSION: Our findings demonstrate that MMF can act as a novel enhancer of mucin production in OMEC in vitro. It has the potential to improve dry eye in patients undergoing OMEC transplantation for bilateral OSD.

Identification of High-Risk Aberrant Crypt Foci and Mucin-Depleted Foci in the Human Colon With Study of Colon Cancer Stem Cell Markers.

BACKGROUND: During colonoscopic screening, only macroscopic lesions will be identified, and these are usually the result of multiple genetic abnormalities. Magnification endoscopic detection of aberrant crypt foci (ACF), long before they acquire complex genetic abnormalities, is promising. However, the features of high-risk ACF-like lesions need to be identified. MATERIALS AND METHODS: In the present cross-sectional study, grossly visible normal mucosal flaps were shaved from 152 colectomies, including 96 colorectal cancer (CRC) cases and 56 controls (22 control specimens with disease with malignant potential and 34 without malignant potential). Methylene and Alcian blue stains were performed directly on the unfixed mucosal flaps to identify ACF and mucin-depleted foci (MDF). Detailed topographic analyses, with immunohistochemical staining for ß-catenin and cancer stem cell (CSC) markers (CD44, CD24, and CD166) were performed. RESULTS: ACF, MDF, and ß-catenin-accumulated crypts were detected more in specimens with adjacent CRC. The left colon had ACF with a larger diameter and greater crypt multiplicity, density, and gyriform pit pattern and were considered the high-risk ACF group. MDF, more commonly associated with dysplasia, is also a marker of possible carcinogenesis. The CD44 CSC marker was significantly upregulated in ACF specimens compared with normal controls. Our 3-tier ACF-only pit pattern classification system showed better linearity with mucosal dysplasia than did the 6-tier Kudo classification. CONCLUSION: High-risk ACF, when detected during chromoendoscopic screening, should be followed up. CSCs might play an important role in pathogenesis. Larger studies and genotypic risk stratification for definite identification of high-risk ACF are needed.