Double-chamber System Research Articles

Degradation of Azo-dye (Disperse Red) Using Rhizosphere Bacterial Consortium

This study investigates the degradation of the azo dye (Disperse Red) using a rhizosphere bacterial consortium. Standard microbiological and molecular techniques were employed to isolate and identify organisms from rhizosphere soil. Degradation of azodye was carried out in a fabricated anoxic and oxic chambers with hydraulic retention time of 40hrs. Initial identification of the bacterial isolates through Gram’s reaction and biochemical tests revealed the presence of organisms belonging to the genera Pseudomonas, Lysinibacillus, and Citrobacter. Molecular and phylogenetic analyses confirmed the isolates as Pseudomonas aeruginosa, Lysinibacillus sphaericus, Pseudomonas chengduensis, and Citrobacter freundii. During the preliminary testing, the degradation efficiency was assessed under varying glucose concentrations. Higher decolorization rate of 56.17% was observed in the medium with 10% glucose after 72 hours, while the medium with 5% glucose achieved a 44.17% colour reduction. Notably, lower degradation rates recorded were 11.96% and 12.85% for the 5% and 10% dye enhance glucose mineral salt media, respectively. However, During the actual degradation testing in a double-chamber system enhanced with biochar, the first anaerobic cycle achieved a maximum decolorization of 71.95% after 94 hours, with the first aerobic cycle further enhancing degradation to 90.51%. The second anaerobic cycle increased degradation to 94.78%, and the final aerobic cycle achieved a decolorization of 98.47%. These results show that the rate of Disperse Red degradation is highly dependent on glucose levels and alternating anaerobic-aerobic conditions. This study demonstrates the potential of using rhizosphere bacterial consortia to bioremediate wastewater contaminated with azo dyes, offering an efficient and sustainable method of environmental management. The results underline the need of optimizing ambient conditions to increase microbial degradation processes.

Simultaneous removal of tetracycline hydrochloride and hexavalent chromium by heterogeneous Fenton in a photocatalytic fuel cell system

In this work, a novel double-chamber system (PFC-Fenton), combined photocatalytic fuel cell (PFC) with Fenton, was constructed for tetracycline hydrochloride (TCH) and hexavalent chromium (Cr(VI)) removal and electricity production. Therein, Zn5(OH)6(CO3)2/Fe2O3/BiVO4/fluorine-doped SnO2 (ZIO/BiVO4/FTO) and carboxylated carbon nanotubes/polypyrrole/graphite felt (CCNTs/Ppy/GF) were served as photoanode and cathode, respectively. Under light irradiation, the removal efficiencies of TCH and Cr(VI) with the addition of H2O2 (2 mL) could reach 93.1% and 80.4%, respectively. Moreover, the first-order kinetic constants (7.37 × 10−3 min−1 of TCH and 3.94 × 10−3 min−1 of Cr(VI)) were 5.26 and 5.57 times as much as the absence of H2O2. Simultaneously, the maximum power density could be obtained 0.022 mW/cm2 at a current density of 0.353 mA/cm2. Therein, the main contribution of TCH degradation was ·OH and holes in anode chamber. The synergistic effect of photoelectrons, generated ·O2−, and H2O2 played a crucial role in the reduction of Cr(VI) in cathode chamber. The high-performance liquid chromatography-mass spectrometry indicated that TCH could be partially mineralized into CO2 and H2O. X-ray photoelectron spectroscope and X-ray absorption near-edge structure spectra showed that Cr(VI) could be reduced to Cr(III). After 5 times of cycling, the removal efficiencies of TCH and Cr(VI) were still greater than 70%, indicating the remarkable stability of the PFC-Fenton system. Overall, this system could remove TCH/Cr(VI) and generate power simultaneously without iron sludge formation, demonstrating a promising method to further develop PFC-Fenton technology.

Biochar based cathode enriched with hydroxyapatite and Cu nanoparticles boosting electromethanogenesis

Electromethanogenesis is an innovative technology addressing the need of storing renewable energy from unprogrammable sources. It allows for the electrochemical production of methane from CO2-rich wastes on microbial cathodes, in a logic of power-to-gas (BEP2G). The challenge of cost-effective and sustainable biocathodes enhancing the microorganism performance and yield of electromethanogenesis is approached in this work. For the first time, porous carbonaceous cathodes were functionalized with Cu nanoparticles and hydroxyapatite (HAP) and successfully experimented for supporting microbial CO2 reduction reaction (CO2RR) to methane. Tests were performed in a double chamber system under CO2 flow at 45 °C. Next Generation Sequencing of 16S RNA indicated that the microbial pool on the cathodes was mostly enriched in Metanobacteriaceae (hydrogenotrophic Archaea) and different fermenting bacteria, depending on the cathode type. High methane production on cathodes made of Cu 20%, HAP 10%, and carbon balance (20Cu/10HAP) was achieved, with a maximum of 866 ± 199 mmol m−2 d−1 (projected cathode area, Coulombic efficiency of 64%), corresponding to values comparable to the maximum in literature, but in shorter timespans (8 vs. 30 days). The documented effect of pH stabilization in the cathodic chamber by HAP was one of the main parameters that concurred to the selectivity of CO2RR towards methane.

Application of a novel gas phase synthesis approach to carbonyl complexes of accelerator-produced 5d transition metals

Abstract In 2014 the first synthesis of a transactinide carbonyl complex – seaborgium hexacarbonyl – was reported. This was achieved in gas-phase chemical experiments in a beam-free environment behind the recoil separator GARIS. Extending this work to heavier elements requires more efficient techniques to synthesize carbonyl complexes as production rates of transactinide elements drop with increasing atomic number. A novel approach was thus conceived, which retains the benefit of a beam-free environment but avoids the physical preseparation step. The latter reduces the yields for products of asymmetric reactions such as those used for the synthesis of suitable isotopes of Sg, Bh, Hs and Mt. For this a series of experiments with accelerator-produced radioisotopes of the lighter homologues W, Re and Os was carried out at the tandem accelerator of JAEA Tokai, Japan. A newly developed double-chamber system, which allows for a decoupled recoil ion thermalization and chemical complex formation, was used, which avoids the low-efficiency physical preseparation step. Here, we demonstrate the feasibility of this newly developed method using accelerator-produced short-lived radioisotopes of the 5d homologues of the early transactinides.

Microbial fuel cells constructed from the integration of anaerobic and aerobic sanitary landfill site systems

The world’s main source of energy now is fossil fuels but the demand for power is increasing. In addition, the burning of fossil fuels produces harmful greenhouse gases and has a significant impact on the environment. The fuel cell system in this study is based on the aerobic and anaerobic integration system are used in most waste disposal methods in Malaysia. It is a system of electrochemistry results from the oxidation of organic matter that transfers electrons to carbon graphite This research is to study the effectiveness of the method generating electricity from micro-fuel cells produced from leachate wastewater and it is also conducted to identify microbial activity using a double chamber system. The food waste obtain is divided into double chambers which is aerobic and anaerobic. Digital readings using a multimeter are performed for ten to thirteen days continuously to obtain the highest reading results for voltage and electric current. The measurement of the highest reading result on the 11th day recorded a reading as high as 146.8 mV at 2000 mV while the current reached 28 μA at 2000 μA. This study has proven that there is the production of electrical sources from the activity of organisms present in food waste using microbial fuel cell systems. The result show that food waste and cattle manure produce the highest voltage and current. This has provided an opportunity to explore alternative ways of generating electricity according to the environment and conditions of each region.

Staged and efficient removal of tetracycline and Cu2+ combined pollution: A designed double-chamber electrochemistry system using 3D rGO

In this study, a staged and efficient advanced treatment process for combined antibiotics and heavy ion pollution based on a designed electrochemical double-chamber system using three-dimensional reduced graphene as the electrode material was established. Tetracycline (TC) and copper ions as the common pollutants in the tailwater of livestock wastewater were chosen as representative antibiotics and heavy ions. A high removal efficiency of almost 100% of tetracycline and copper ions at low concentrations (10 mg/L) was obtained using the electrochemical double-chamber system. Under the optimal reaction conditions, the estimated energy consumption was 0.25 KWhm−3. Among the 20 mg/L and 50 mg/L composite pollutants, the removal efficiencies of tetracycline were 92% and 82%, respectively, and the removal efficiencies of copper ions were 99% and 100%, respectively. In the treatment of actual tailwater from livestock wastewater, the removal efficiencies of copper ions and total organic carbon (TOC) were 89% and 63%, respectively. In the same treatment time, compared to a single-chamber reaction system, the TC removal efficiencies increased by 13%, 20%, and 26%, and the removal efficiencies of Cu2+ increased by 28%, 48%, and 41% in the treatment of 10 mg/L, 20 mg/L, and 50 mg/L composite pollutants, respectively. In addition, the staged disposal method based on the electrochemical double-chamber system had a higher efficiency and double the treatment capacity for removing combined tetracycline and copper ion pollution than a general electrochemistry system in an equal time period. This study offers a practical and efficient treatment system for combined pollution in water.

A Dual Zinc plus Arginine formulation protects against tumor necrosis factor-alpha-induced barrier dysfunction and enhances cell proliferation and migration in an in vitro gingival keratinocyte model

ObjectiveTo investigate the effects of Dual Zinc plus Arginine formulations (aqueous solution and dentifrice) on tumor necrosis factor-alpha (TNF-α)-induced barrier dysfunction as well as on cell proliferation and migration in an in vitro gingival keratinocyte model. DesignGingival keratinocytes were seeded onto the membrane of a double-chamber system in the absence and presence of recombinant TNF-α and the formulations under investigation. The barrier function was assessed by determination of transepithelial electrical resistance (TER) and paracellular transport of fluorescein isothiocyanate (FITC)-dextran. The distribution of zonula occludens-1 (ZO-1) and occludin was visualized by immunofluorescence microscopy. The effects of the formulations on keratinocyte cell proliferation were determined using a fluorescent cell tracker dye, while a migration assay kit was used to investigate their effects on cell migration. ResultsUnder conditions where TNF-α induces loss of keratinocyte barrier integrity, the Dual Zinc plus Arginine formulations (aqueous solution and dentifrice) protected the keratinocyte tight junction against the damages since they prevented the TNF-α-induced drop in TER and increase in FITC-dextran paracellular flux in the in vitro model. The treatment of keratinocytes with the formulations markedly mitigated the altered distribution of ZO-1 and occludin. Both formulations increased the proliferation of keratinocytes and alleviated the negative impact caused by TNF-α. Lastly, the formulations increased the migration capacity of keratinocytes. ConclusionsThe ability of the Dual Zinc plus Arginine formulations to protect the barrier integrity of gingival keratinocytes from TNF-α-induced damage and to promote their proliferation and migration suggests that they may offer benefits for oral health.

BULGUR INDUSTRY WASTEWATER TREATMENT BY MICROBIAL FUEL CELL – EXPLORATORY STUDY

The bulgur industry has importance in the food sector in Turkey. The wastewater generated from this industry can be considered bio waste. The microbial fuel cells (MFC) are a relatively new technique aiming to treat the wastewater and producing direct energy. This study aims to explore the degradation efficiency of the organic matters expressed as chemical oxygen demand (COD) founded in bulgur industry wastewater by microbial fuel cell techniques. Furthermore, it aims to study the potential formation of electricity from this type of wastewater. In this study, the MFC – double chamber system was performed. 1.5 L bulgur industry wastewater containing 3% of biomass was used. The COD and the voltage were measured. The COD generated from the bulgur industry wastewater was 28800 mg/L. After using the MFC system, the COD was decreased to reach 2560 mg/L with a removal efficiency of 91%. 1st order kinetic model had the best fit for COD removal with a correlation coefficient (R2) of 0.95. The maximum and average voltages were 0.448 Volt and 0.180 Volt, respectively. The average voltage for every 1 m2 was 45 volt. As a result of the exploratory study, the MFC can be used to treat the bulgur industry wastewater and generating energy. But it should be combined with other treatment methods tomeet the COD standard limits.

Abstract 275: A possible relationship between gene mutation profiles and activity of E7386 in in vitro models of colorectal cancer organoids

Abstract A novel beta-catenin/CBP binding modulator E7386 is expected to affect colorectal carcinoma (CRC) cells particularly those with aberrant activation of Wnt/beta-catenin signaling pathway. The Wnt signaling pathway is recently reported to play important roles for activation of not only CRC cells but also cancer-associated fibroblasts (CAFs). Here, we established an in vitro model of CRC-derived organoids as well as a co-culture method of the CRC organoids with CAFs, and effects of E7386 on the growth of the organoids were examined. Fragments of CRC tissues from a total of 45 patients who underwent surgery were obtained, dissociated into single cells, which were seeded on polymerized Matrigel (Corning), and cultured with serum-free media containing EGF and Noggin optimized for organoids using 12-well plates. Fourteen CRC organoids from 45 cases were successfully established. For 2D culture of CAFs from CRC tissues of each patient, RPMI medium containing 10% FBS was used. A co-culture method of the CRC organoids with CAFs was applied using a double chamber system with transwell inserts in 24-well plates. Cell viabilities of the organoids and CAFs were measured using CellTiter-Glo Luminescent Cell Viability Assay reagent (Promega). Gene aberrations in original CRC tissues were analyzed using NCC Oncopanel, and gene expression profiles in established CRC organoids and CAFs were analyzed using SurePrint G3 Human GE microarray (Agilent). [Effects of CAFs on cell viabilities and gene expressions of organoids] Cell viabilities of organoids and CAFs appeared to increase at 72 hours after beginning of the culture by 2 to 8 and 1 to 2-fold, respectively. When organoids were co-cultured with CAFs, cell viabilities of organoids were increased and epithelial-mesenchymal transition-related gene expressions were dramatically changed in several cases, but not in all cases. The organoids co-cultured with CAFs appeared to be more sensitive to E7386 as compared to those cultured alone. [Possible relationship between gene mutations in CRCs and activity of E7386 in CRC organoids] Cell viabilities of organoids without APC mutations (n=1) and organoids with APC and TP53 mutations alone (n=2) were less than those of organoids with other oncogene mutations, e.g., KRAS or PIK3CA (n=4). Efficacy of E7386 on the cell viabilities of organoids with APC and TP53 mutations alone was found at lower concentrations at 30-100 nM, as compared to those of organoids without APC mutations. From the results, CRC organoids were established from 14 patients, and activity of E7386 appeared to be related to gene mutation profiles in the established CRC organoids. Citation Format: Toshio Imai, Mie Naruse, Masako Ochiai, Yoichi Ozawa, Takashi Owa, Atsushi Ochiai. A possible relationship between gene mutation profiles and activity of E7386 in in vitro models of colorectal cancer organoids [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 275.

Performance of microbial fuel cell double chamber using mozzarella cheese whey substrate

Nowadays the availability of electric energy is decreasing, hence there is a need for innovation of electric energy producer alternative; one of them is microbial fuel cell (MFC). MFC is a bioelectrochemical system generated by bacterial metabolism that utilizes organic substrate. One of the substrates that can be used is whey, a waste generated from cheese production. Therefore, this study aimed to determine the power of potential current and voltage generated from the use of whey cheese as a substrate for bacterial metabolism. In this research, double chamber system was used in microbial fuel cell reactor by using cheese whey as substrate at anode and potassium permanganate as cathode and utilizing membrane nafion 212 as membrane of proton exchange. The variable of experiment was bacteria type. The types of bacteria used in this study were Lactobacillus bulgaricus, Streptococcus thermophillus and Lactobacillus casei. While the operating time used was 100 hours. The highest current produced was 74.6 μA and the highest voltage was 529.3 mV produced by Lactobacillus bulgaricus bacteria. In this study, it was also found that the death phase of the three bacteria was at 70-80 hours.

Single chamber microbial fuel cell (SCMFC) with a cathodic microalgal biofilm: A preliminary assessment of the generation of bioelectricity and biodegradation of real dye textile wastewater

An air exposed single-chamber microbial fuel cell (SCMFC) using microalgal biocathodes was designed. The reactors were tested for the simultaneous biodegradation of real dye textile wastewater (RTW) and the generation of bioelectricity. The results of digital image processing revealed a maximum coverage area on the biocathodes by microalgal cells of 42%. The atmospheric and diffused CO2 could enable good algal growth and its immobilized operation on the cathode electrode. The biocathode-SCMFCs outperformed an open circuit voltage (OCV), which was 18%–43% higher than the control. Furthermore, the maximum volumetric power density achieved was 123.2 ± 27.5 mW m−3. The system was suitable for the treatment of RTW and the removal/decrease of COD, colour and heavy metals. High removal efficiencies were observed in the SCMFCs for Zn (98%) and COD (92–98%), but the removal efficiencies were considerably lower for Cr (54–80%). We observed that this single chamber MFC simplifies a double chamber system. The bioelectrochemical performance was relatively low, but the treatment capacity of the system seems encouraging in contrast to previous studies. A proof-of-concept experiment demonstrated that the microalgal biocathode could operate in air exposed conditions, seems to be a promising alternative to a Pt cathode and is an efficient and cost-effective approach to improve the performance of single chamber MFCs.

Similar Sealability Between Bioceramic Putty Ready-To-Use Repair Cement and White MTA

This study was designed to assess the ability to prevent glucose penetration of a bioceramic putty ready-to-use repair cement comparing to white mineral trioxide aggregate (WMTA). After root canal instrumentation, the apical 3 mm of maxillary incisors were resected and root-end cavities with depth of 3 mm were prepared with ultrasound and filled with the tested materials (15 roots per group). All roots were mounted in a double chamber system to assess glucose penetration using 15 psi pressure application. After 1 h, glucose concentrations in the lower chamber were measured following an enzymatic reaction. Four roots were used as controls. The Mann-Whitney test verified differences in glucose leakage between groups and the Tukey's test was used for multiple comparisons. Significance level was set at 5%. There was no significant difference in glucose leakage between iRoot BP Plus and White MTA groups. iRoot BP Plus had a similar ability to that of white MTA in preventing glucose leakage as a root-end filling material.

Microarray analysis of gene expression of bone marrow stem cells cocultured with salivary acinar cells

Our previous work has demonstrated that rat bone marrow stem cells (BMSCs) can transdifferentiate into α-amylase-producing cells after coculture with rat submandibular gland acinar cells. These transdifferentiated cells may be used for regeneration of damaged salivary gland. The purpose of this study was to investigate the global gene expression of rat BMSCs cocultured with rat submandibular gland acinar cells and the factors inducing this transdifferentiation. Rat BMSCs were indirectly cocultured with rat submandibular gland acinar cells by using the double chamber system for 5 and 10 days. The global gene expression of BMSCs during transdifferentiation into acinar cells was investigated by microarray analysis. A total of 45,018 probes were used and 41,012 genes were detected. After coculture for 5 days, 1409 genes were upregulated more than twofold and 1417 genes were downregulated more than twofold (p<0.005). Moreover, after coculture for 10 days, 1356 genes were upregulated more than twofold and 1231 genes were downregulated more than twofold (p<0.005). Bone morphogenetic protein (BMP)-6 was one of the top-ranked upregulated genes. The hub genes were interleukin-6 and CCAAT/enhancer-binding protein β (CEBPB) in the early and late response gene groups, respectively. This is believed to be the first study on the global gene expression of rat BMSCs cocultured with rat acinar cells. Many genes related to the function of salivary acinar cells such as those responsible for the production of α-amylase protein were upregulated and many genes related to the differentiation of BMSCs into adipocytes and osteoblasts were downregulated. In addition, BMP-6 gene was found to be highly upregulated. We proposed that three target genes, BMP-6, interleukin-6 and CEBPB, play important roles in the transdifferentiation of BMSCs into acinar cells, and are worthy of further investigation.

Transdifferentiation of Human Amniotic Epithelial Cells into Acinar Cells Using a Double-Chamber System

This study investigated the transdifferentiation of stem cells from human amnion tissue into functional acinar cells (ACs) using a co-culture system. Human amniotic epithelial cells (hAECs) were isolated from amnion tissue by mechanical mincing and enzymatic digestion. After primary culture, the phenotype of the cells was identified by flow cytometry (FCM) and immunocytochemical staining. hAECs were co-cultured with submandibular gland acinar cells of SD rats using a double-chamber system. The expression of α-amylase was determined by immunocytochemical method and fluorescent real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) after induction for 1 and 2 weeks, respectively. Digestion with trypsin is an effective method for isolating hAECs from amnion tissue. These cells were positive for CD29 and CK19 and weakly positive for CD44 and α-amylase. Within 2 weeks, α-amylase in hAECs increased with induction time. The expression of α-amylase in hAECs was increased 3.38-fold after co-culturing for 1 week. This ratio increased to 6.6-fold, and these cells were positive for mucins, after co-culturing for 2 weeks. hAECs possess the potential to differentiate into ACs in vitro. They might be a stem cell resource for clinical applications of cell replacement therapy in salivary gland dysfunction diseases.

Entry of Herpes Simplex Virus Type 1 (HSV-1) into the Distal Axons of Trigeminal Neurons Favors the Onset of Nonproductive, Silent Infection

Following productive, lytic infection in epithelia, herpes simplex virus type 1 (HSV-1) establishes a lifelong latent infection in sensory neurons that is interrupted by episodes of reactivation. In order to better understand what triggers this lytic/latent decision in neurons, we set up an organotypic model based on chicken embryonic trigeminal ganglia explants (TGEs) in a double chamber system. Adding HSV-1 to the ganglion compartment (GC) resulted in a productive infection in the explants. By contrast, selective application of the virus to distal axons led to a largely nonproductive infection that was characterized by the poor expression of lytic genes and the presence of high levels of the 2.0-kb major latency-associated transcript (LAT) RNA. Treatment of the explants with the immediate-early (IE) gene transcriptional inducer hexamethylene bisacetamide, and simultaneous co-infection of the GC with HSV-1, herpes simplex virus type 2 (HSV-2) or pseudorabies virus (PrV) helper virus significantly enhanced the ability of HSV-1 to productively infect sensory neurons upon axonal entry. Helper-virus-induced transactivation of HSV-1 IE gene expression in axonally-infected TGEs in the absence of de novo protein synthesis was dependent on the presence of functional tegument protein VP16 in HSV-1 helper virus particles. After the establishment of a LAT-positive silent infection in TGEs, HSV-1 was refractory to transactivation by superinfection of the GC with HSV-1 but not with HSV-2 and PrV helper virus. In conclusion, the site of entry appears to be a critical determinant in the lytic/latent decision in sensory neurons. HSV-1 entry into distal axons results in an insufficient transactivation of IE gene expression and favors the establishment of a nonproductive, silent infection in trigeminal neurons.

Comparison of the root-end seal provided by bioceramic repair cements and White MTA

To compare the ability of Ceramicrete, BioAggregate and white ProRoot MTA (mineral trioxide aggregate) to prevent glucose leakage through root-end fillings. After root canal instrumentation, the apical 3 mm of maxillary incisors were resected and retropreparations, 3 mm in depth, were created with ultrasound. Root-end cavities were filled with the tested materials (15 roots per group). All roots were mounted in a double-chamber system to assess glucose penetration using 15 psi pressure application. After 1 h, glucose concentrations in the lower chamber were measured following an enzymatic reaction. Four roots were used as controls. One-way anova verified differences in glucose leakage between groups and Tukey test performed multiple comparisons. Significance was set at α = 5%. There was a significant difference between the three materials (anova, P < 0.05). Ceramicrete had significantly lower glucose penetration than BioAggregate (Tukey, P < 0.05). There was no difference between the two bioceramic cements and white MTA (P > 0.05). Both endodontic bioceramic repair cements displayed similar leakage results to white MTA when used as root-end fillings materials. Ceramicrete had significantly lower glucose penetration.

Ovarian cancer cells stimulate uPA gene expression in fibroblastic stromal cells via multiple paracrine and autocrine mechanisms

ObjectivesExpression of uPA mRNA is massively up-regulated in the stroma of poorly differentiated ovarian tumors. We hypothesized that this expression was induced by paracrine signals from the epithelial tumor cells, and established an in vitro model of ovarian cancer microenvironment to study intercellular cross-talk. MethodsES-2 clear cell carcinoma cells were grown in tissue culture inserts in a double-chamber system with fibroblastic stromal LEP cells embedded in Matrigel. Binding-site directed antibodies were used to neutralize soluble cytokines in ES-2 conditioned medium (CM) before incubation with LEP cells. Real time PCR measured uPA mRNA in LEP cells, as well as mRNA for cytokines in both cell types. ResultsCo-culture with ES-2 cells as well as incubation with ES-2 CM induced uPA mRNA in LEP cells about two-fold. In short time (12 h) incubation of LEP cells with CM, antibodies to EGF and bFGF reduced induction of uPA mRNA, suggesting that these cytokines function as paracrine signals. EGF mRNA and bFGF mRNA were also found in ES-2 cells. At longer incubation (24 h) antibodies to bFGF, HB-EGF, HGF, IGF-1, and IL-1α reduced uPA mRNA induction, suggesting an autocrine function for these cytokines in LEP cells. In fact, expression of the same five cytokines was up-regulated in LEP cells exposed to CM. ConclusionWe identified two cytokines as paracrine signals, and five cytokines as autocrine signals in ovarian cancer cell induced up-regulation of uPA mRNA in stromal fibroblastic cells. It is crucial to understand intra-tumoral cross-talk, since it can offer new therapeutic approaches.

Antigen-presenting capacity of macrophages and dendritic cells in the peritoneal cavity of patients treated with peritoneal dialysis.

In this study the antigen-presenting capacity of human peritoneal cells and the influence of continuous ambulant peritoneal dialysis (CAPD) were studied. On average 6% of the peritoneal cells were dendritic cells (DC), with no difference between CAPD and control peritoneal cells. DC were enriched by selecting for non-adherent, Fc receptor-negative, low density cells. A typical spot-like CD68 positivity was seen in DC, in contrast to the pancytoplasmic staining pattern in macrophages. Peritoneal DC morphologically and functionally showed features of cells belonging to the DC lineage. Peritoneal DC were superior antigen-presenting cells for both allo-antigen, and Candida albicans antigen or purified protein derivative. CAPD peritoneal macrophages were two- to three-fold better stimulator cells for allogeneic T cells compared with control macrophages. The level of integrins/adhesins or MHC class I or II, as measured semi-quantitatively on the FACS, could not account for this phenomenon. In addition, a double chamber system showed that dialysate-activated macrophages produced soluble factors that could enhance DC-induced allogeneic T cell proliferation. In conclusion, human peritoneal cells contain a relatively high percentage of classical DC. CAPD treatment does not impair the antigen-presenting capacity of peritoneal cells, but instead upregulates the allo-antigen-presenting capacity of peritoneal macrophages.

Morphological and Behavioural Features of Bewo Cells Grown on Matrigel Offers a Model for Human Cytotrophoblast Cells During Early Implantation

In order to observe the morphological changes that occur in cell-cell and cell-matrix interactions among trophoblast cells and other neighbouring cells during human implantation, BeWo cells were cultured in a 3-dimensional artificial extracellular matrix (Matrigel) using a double-chamber system. These cultures were then processed for light and electron microscopical examination. Results obtained show that the morphology and behavioural pattern of BeWo cells grown on Matrigel is similar to that reported for in vitro normal human cytotrophoblast cells on extracellular matrix (ECM). These results suggest that culture of BeWo cells on extracellular matrices may be useful for the study of some of the early embryonic events leading to human implantation, especially during the period when trophoblastic cells interact with and erode the uterine epithelium and ECM. Keywords: BeWo cells, Trophoblast, Morphology, Matrigel, implantation Journal of Science and Technology Vol. 28 (1) 2008 pp. 4-16

Transdifferentiation of Bone Marrow Stem Cells into Acinar Cells Using a Double Chamber System

Hypofunction of the salivary glands can substantially affect quality of life. Current treatments for salivary hypofunction are of limited effectiveness. Although the implantation of functional salivary gland tissue from autologous glandular cells represents a possible physiologic solution to this problem, tissue engineering of salivary glands would require the generation of a great number of acinar cells (ACs). The purpose of this study was to investigate the feasibility of transdifferentiation of bone marrow stem cells (BMSCs) into functional ACs using a co-culture system. BMSCs were isolated from adult rats and co-cultured with rat parotid ACs using a double chamber system. The transdifferentiation of BMSCs was evaluated by immunocytochemical analysis of alpha-amylase, which has unique functional expression in ACs. Expression of alpha-amylase, indicating successful transdifferentiation of BMSCs into ACs, was found in 30% of BMSCs after co-culturing for 1 week, and in 50% after co-culturing for 2 and 3 weeks. This study has demonstrated the potential of rat BMSCs to transdifferentiate into ACs, and support the feasibility of application of BMSCs in salivary gland tissue engineering.