133 results on '"Costa, R. H."'
Search Results
2. An N-terminal inhibitory domain modulates activity of FoxM1 during cell cycle
- Author
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Park, H J, Wang, Z, Costa, R H, Tyner, A, Lau, L F, and Raychaudhuri, P
- Published
- 2008
- Full Text
- View/download PDF
3. Effect of COD/N ratio on N2O production during nitrogen removal by aerobic granular sludge.
- Author
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Magnus, B. S., Daudt, G. C., Xavier, J. A., Guimarães, L. B., Costa, R. H. R., and Velho, V. F.
- Subjects
SLUDGE management ,NITROGEN removal (Sewage purification) ,NITROUS oxide ,CHEMICAL oxygen demand ,NITRIFICATION - Abstract
N
2 O-production was investigated during nitrogen removal using aerobic granular sludge (AGS) technology. A pilot sequencing batch reactor (SBR) with AGS achieved an effluent in accordance with national discharge limits, although presented a nitrite accumulation rate of 95.79% with no simultaneous nitrification–denitrification. N2 O production was 2.06 mg L−1 during the anoxic phase, with N2 O emission during air pulses and the aeration phase of 1.6% of the nitrogen loading rate. Batch tests with AGS from the pilot reactor verified that at the greatest COD/N ratio (1.55), the N2 O production (1.08 mgN2 O-N L−1 ) and consumption (up to 0.05 mgN2 O-N L−1 ), resulted in the lowest remaining dissolved N2 O (0.03 mgN2 O-N L−1 ), stripping the minimum N2 O gas (0.018 mgN2 O-N L−1 ). Conversely, the carbon supply shortage, under low C/N ratios, increased N2 O emission (0.040 mgN2 O-N L−1 ), due to incomplete denitrification. High abundance of ammonia-oxidizing and low abundance of nitrite-oxidizing bacteria were found, corroborating the fact of partial nitrification. A denitrifying heterotrophic community, represented mainly by Pseudoxanthomonas, was predominant in the AGS. Overall, the AGS showed stable partial nitrification ability representing capital and operating cost savings. The SBR operation flexibility could be advantageous for controlling N2 O emissions, and extending the anoxic phase would benefit complete denitrification in cases of low C/N influents. [ABSTRACT FROM AUTHOR]- Published
- 2017
- Full Text
- View/download PDF
4. Ecology of duckweed ponds used for nutrient recovery from wastewater.
- Author
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Teles, C. C., Mohedano, R. A., Tonon, G., Filho, P. Belli, and Costa, R. H. R.
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DUCKWEEDS ,SEWAGE ,WATER purification ,ZOOPLANKTON ,NUCLEOTIDE sequencing - Abstract
The microorganism community that grows under duckweed shelter can play an important role on treatment processes. Therefore, the present study aimed to assess the zooplankton dynamic and microbial community in duckweed ponds (DPs) applied for domestic wastewater treatment under open field conditions. A pilot system comprised of two DPs in series (DP1 and DP2), with 10 m² each, received domestic wastewater through a flow rate of 200 L⋅day
-1 . Thus, the system was monitored during 314 days through samples collected and analysed weekly. Also, the zooplankton organisms were identified and quantified. DNA sequencing was performed in order to identify the bacterial populations. The findings showed a high efficiency of nutrient removal with 93% and 91% of total phosphorus and total nitrogen, respectively. A high density of microcrustaceans was observed in DP1 reaching 4,700 org.100 mL-1 and rotifers (over than 32,000 org.100 mL-1 ) in DP2, that could be related to the low suspended solids concentration (<30 mg⋅L-1 ) and turbidity (<10 NTU). The bacterial community showed a strong heterogeneity between samples collected along the seasons. Through these findings, it is possible to realise that the understanding of ecology could help to enhance the operation and designs of DPs. [ABSTRACT FROM AUTHOR]- Published
- 2017
- Full Text
- View/download PDF
5. Valuation of OSA process and folic acid addition as excess sludge minimization alternatives applied in the activated sludge process.
- Author
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Martins, C. L., Velho, V. F., Ramos, S. R. A., Pires, A. S. C. D., Duarte, E. C. N. F. A., and Costa, R. H. R.
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SLUDGE management ,FOLIC acid ,CHEMICAL oxygen demand ,WATER purification ,WATER pollution - Abstract
The aim of this study was to investigate the ability of the oxic-settling-anaerobic (OSA)-process and the folic acid addition applied in the activated sludge process to reduce the excess sludge production. The study was monitored during two distinct periods: activated sludge system with OSAprocess, and activated sludge system with folic acid addition. The observed sludge yields (Y
obs ) were 0.30 and 0.08 kg TSS kg-1 chemical oxygen demand (COD), control phase and OSA-process (period 1); 0.33 and 0.18 kg TSS kg-1 COD, control phase and folic acid addition (period 2). The Yobs decreased by 73 and 45% in phases with the OSA-process and folic acid addition, respectively, compared with the control phases. The sludge minimization alternatives result in a decrease in excess sludge production, without negatively affecting the performance of the effluent treatment. [ABSTRACT FROM AUTHOR]- Published
- 2016
- Full Text
- View/download PDF
6. Cytokine regulation of the liver transcription factor hepatocyte nuclear factor-3β is mediated by the C/EBP family and interferon regulatory factor 1
- Author
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Samadani, U., Porcella, A., Pani, L., Peter Johnson, Burch, J. B. E., Pine, R., and Costa, R. H.
- Published
- 1995
7. Aerobic granular sludge technology and nitrogen removal for domestic wastewater treatment.
- Author
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Wagner, J., Guimarães, L. B., Akaboci, T. R. V., and Costa, R. H. R.
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AERATED package treatment systems ,NITROGEN removal (Sewage purification) ,NITRIFICATION ,SEWAGE purification ,TECHNOLOGICAL obsolescence - Abstract
This study evaluated aerobic granulation and nitrogen removal via assimilation, nitrification, and denitrification of a system fed with real domestic wastewater. The granulation process was complete after 160 days of operation. The mature granules had an almost spherical structure, an average size of 473.0 μm, and a good settling ability (SVI
30 of 75.6 mL g-1 ). Ammonium assimilation for cell growth varied between 3.5 and 64.6% during reactor start-up. After granule formation, assimilation accounted for less than 5% and nitrogen was mainly removed by partial nitrification up to nitrite, followed by denitrification via nitrite. Average efficiencies of 86.6% for nitrification, 59.5% for denitrification, and 60.5% for total nitrogen were obtained in this period. The assimilation ability of the mature granules grown on domestic wastewater was lower than the commonly reported results obtained for synthetic granules. [ABSTRACT FROM AUTHOR]- Published
- 2015
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- View/download PDF
8. Consortia of microalgae and bacteria in the performance of a stabilization pond system treating landfill leachate.
- Author
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Costa, R. H. R., Martins, C. L., Fernandes, H., and Velho, V. F.
- Subjects
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MICROALGAE , *BACTERIA , *LEACHATE , *SANITARY landfills , *SEWAGE lagoons - Abstract
This study treated sanitary landfill leachate and was conducted in a pilot-scale system composed of three serial ponds (P1, P2 and P3), followed by a rock filter, in order to evaluate the microbial consortium influence on system performance and to investigate microorganism dynamics in the process. The system was broken into three stages, with a continuous flow rate (Q = 200 L d-1) for 43 weeks. The stages were as follows: conventional operation (stage I), 12 h aeration in P2 (stage II), and 18 h aeration in P2 (stage III). The results showed the possibilities for treating landfill leachate, presenting an average efficiency of 75% for both filtered biochemical oxygen demand and ammonium. At the end of stage III, the ammonium concentration was 6 mg L-1, which is lower than that established by Brazilian regulations for wastewater discharge (CONAMA 430/2011). The aeration applied in P2 led to a change in the microbial consortia during the second and third stage, which influenced the quality of the final effluent. The best performance was seen in stage III, where the system showed high microbial diversity, including the presence of nitrifying bacteria. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
9. Using full-scale duckweed ponds as the finish stage for swine waste treatment with a focus on organic matter degradation.
- Author
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Mohedano, R. A., Costa, R. H. R., Hofmann, S. M., and Belli Filho, P.
- Subjects
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WASTE treatment , *BIODEGRADATION of organic compounds , *DUCKWEEDS , *CHEMICAL oxygen demand , *EFFLUENT quality - Abstract
The rapid increase in the number of swine has caused pronounced environmental impacts worldwide, especially on water resources. As an aggregate, smallholdings have an important role in South American pork production, contributing to the net diffusion of pollution. Thus, duckweed ponds have been successfully used for swine waste polishing, mainly for nutrient removal. Few studies have been carried out to assess organic matter degradation in duckweed ponds. Hence, the present study evaluated the efficiency of two full-scale duckweed ponds for organic matter reduction of swine waste on small pig farms. Duckweed ponds, in series, received the effluent after an anaerobic biodigester and storage pond, with a flow rate of 1 m3 day-1. After 1 year of monitoring, an improvement in effluent quality was observed, with a reduction in biochemical oxygen demand (BOD) and total chemical oxygen demand (tCOD), respectively, of 94.8 and 96.7%, operating at a loading rate of approximately 27 kgBOD ha-1 day-1 and 131 kgCOD ha-1 day-1. Algae inhibition due to duckweed coverage was strongly observed in the pond effluent, where chlorophyll a and turbidity remained below 25 μg L-1 and 10 NTU. Using the study conditions described herein, duckweed ponds were shown to be a suitable technology for swine waste treatment, contributing to the environmental sustainability of rural areas. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
10. Performance and Kinectics Aspects of Nitrogen Removal in a Biofilm Sequencing Batch Reactor.
- Author
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Costa, R. H. R., Wolff, D. B., and Souto, V. S.
- Abstract
A biofilm sequencing batch reactor with a volume of 1.42 m
3 , nylon nets providing a 4,140 m2 /m3 support area for biofilms and an automated operation with 8 hour cycles was studied. The duration of the experiment was 135 days. Removal efficiencies e≧ 80% were obtained for carbonaceous matter, producing an effluent with 31±26.8 mg/L of filtered COD, 7±3.6 mg/L of BOD5 and 12±3.2 mg/L of TOC. The average removal efficiency of ammonium was 77 ± 16.6%, with a mean concentration in the effluent of 14 ± 10.2 mg NH4 -N/L. The denitrification efficiency was 80±14.7%. The effluent characteristics met the requirements of Brazilian environmental standard for discharge to receiving water bodies. A kinetic study of nitrification and denitrification showed that during the aerobic phase the specific rate of ammonium consumption was 0.057 g NH4 -N/g VSS.d and the production of NOx-N was 0.074 g NOx-N/g VSS.d, while the specific rate of NOx-N consumption was 0.05 g NOx-N/g VSS.d during the anoxic phase. The suspended and fixed biomass was composed of 50% ammonium-oxidizing bacteria (AOB). [ABSTRACT FROM AUTHOR]- Published
- 2013
11. Nutrient recovery from swine waste and protein biomass production using duckweed ponds (Landoltia punctata) Southern Brazil.
- Author
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Mohedano, R. A., Veiho, V. F., Costa, R. H. R., Hofmann, S. M., and Beth Fliho, P.
- Subjects
ANIMAL waste ,BIOMASS production ,PORK products ,NITROGEN ,PHOSPHORUS - Abstract
Brazil is one of the most important countries in pork production worldwide, ranking third. This activity has an important role in the national economic scenario. However, the fast growth of this activity has caused major environmental impacts, especially in developing countries. The large amount of nitrogen and phosphorus compounds found in pig manure has caused ecological imbalances, with eutrophication of major river basins in the producing regions. Moreover, much of the pig production in developing countries occurs on small farms, and therefore causes diffuse pollution. Therefore, duckweed pond have been successfully used in the swine waste polishing, generating further a biomass with high protein content. The present study evaluated the efficiency of two full scale duckweed ponds for the polishing of a small pig farm effluent, biomass yield and crude protein (CP) content. Duckweed pond series received the effluent from a biodigester-storage pond, with a flow rate of 1 m
3 /day (chemical oxygen demand rate = 186 kg/ha day) produced by 300 animals. After 1 year a great improvement of effluent quality was observed, with removal of 96% of total Kjeldahl nitrogen (TKN) and 89% of total phosphorus (TP), on average. Nitrogen removal rate is one of the highest ever found (4.4 g TKN/m2 day). Also, the dissolved oxygen rose from 0.0 to 3.0 mg/L. The two ponds produced together over 13 tons of fresh biomass (90.5% moisture), with 35% of CP content, which represents a productivity of 24 tonsCP/ha year. Due to the high rate of nutrient removal, and also the high protein biomass production, duckweed ponds revealed, under the presented conditions, a great potential for the polishing and valorization of swine waste. Nevertheless, this technology should be better exploited to improve the sustainability of small pig farms in order to minimize the impacts of this activity on the environment. [ABSTRACT FROM AUTHOR]- Published
- 2012
- Full Text
- View/download PDF
12. Modelling waste stabilisation ponds with an extended version of ASM3.
- Author
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Gehring, T., Silva, J. D., Kehl, O., Castilhos Jr., A. B., Costa, R. H. R., Uhlenhut, F., Alex, J., Horn, H., and Wichern, M.
- Subjects
SEWAGE sludge digestion ,MATHEMATICAL models ,SEWAGE lagoons ,NITROGEN removal (Sewage purification) ,NITROGEN compounds ,LANDFILLS ,CARBON dioxide ,INDUSTRIAL wastes ,SOLID waste - Abstract
In this paper an extended version of IWA's Activated Sludge Model No 3 (ASM3) was developed to simulate processes in waste stabilisation ponds (WSP). The model modifications included the integration of algae biomass and gas transfer processes for oxygen, carbon dioxide and ammonia depending on wind velocity and a simple ionic equilibrium. The model was applied to a pilot-scale WSP system operated in the city of Florianópolis (Brazil). The system was used to treat leachate from a municipal waste landfill. Mean influent concentrations to the facultative pond of 1,456 g
COD /m3 and 505 gNH4-N /m3 were measured. Experimental results indicated an ammonia nitrogen removal of 89.5% with negligible rates of nitrification but intensive ammonia stripping to the atmosphere. Measured data was used in the simulations to consider the impact of wind velocity on oxygen input of 11.1 to 14.4 gO2 /(m2 d) and sun radiation on photosynthesis. Good results for pH and ammonia removal were achieved with mean stripping rates of 18.2 and 4.5 gN /(m2 d) for the facultative and maturation pond respectively. Based on measured chlorophyll a concentrations and depending on light intensity and TSS concentration it was possible to model algae concentrations. [ABSTRACT FROM AUTHOR]- Published
- 2010
- Full Text
- View/download PDF
13. Utilization of a hybrid sequencing batch reactor (HSBR) as a decentralized system of domestic wastewater treatment.
- Author
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Da Costa, R. H. R., Souto, V. S., Prelhaz, A. T. S., Neto, L. G. L., and Wolff, D. B.
- Subjects
- *
DENITRIFICATION , *NITRIFICATION , *SEWAGE , *BIOMASS , *BIOREMEDIATION , *INDUSTRIAL wastes - Abstract
This paper presents the experiments carried out in a hybrid sequencing batch reactor (HSBR), used for biological treatment of sewage. The HSBR was built in a cylindrical shape and made of stainless steel, with a volume of 1.42m³. Besides the biomass in suspension, the reactor also carried fixed biomass (hybrid process), adhered in the support material. This consisted of a nylon net disposed in a grille for biofilm biomass adhesion. The reactor worked fully automated in operational cycles of maximum 8 hours each, presenting the following phases: filling, anoxic, aerobic, settle and draw of treated effluent, with 3 fillings per cycle. Increasing organic loads (0.14 to 0.51 kg TCOD/m³ day) and ammonium loads (0.002 to 0.006 kgNH4-N/m³·day) were tested. We monitored the reactor's performance by measuring the liquid phase (COD, pH, temperature, DO, nitrogen and phosphorus) during the cycles and by measuring the sludge through respirometric tests. The results obtained demonstrated TCOD removal efficiency between 73 and 96%, and ammonium removal efficiency between 50 and 99%. At the end of the cycles, the effluent presented ammonium concentration <20 mg/L, meeting the Brazilian environmental legislation standards (CONAMA 357/2005) regarding discharges into the water bodies. Respirometric tests showed biomass dependency on FCOD concentrations. Results have demonstrated the potential of this type of reactor for decentralized treatment of domestic wastewater. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
14. Evaluation of sludge from pond system for treatment of piggery wastes.
- Author
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Zanotelli, C. T., Costa, R. H. R., and Perdomo, C. C.
- Subjects
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SEWAGE lagoons , *PORK industry , *SEWAGE aeration , *AGRICULTURAL wastes & the environment , *AGRICULTURAL waste recycling , *PHOSPHORUS & the environment , *INDUSTRY & the environment - Abstract
Stabilization ponds used for the treatment of piggery wastes accumulate sludge over time, which is commonly used in agriculture. The objective of this study was to evaluate the agronomic potential of this kind of sludge. The samplings were collected in two different phases. The first in two anaerobic ponds (AP1 and AP2) and in one facultative pond with 5 transverse baffles and, the second in the same facultative pond with aeration. The removed sludge of AP1 and AP2 was characterized as rich sludge in volatile solids and with low stabilization, there was a great accumulation of the total phosphorus in the sludge of AP2. The facultative pond presented greater retention of nutrients in the sludge in relation to the anaerobic ponds. The annual accumulation of sludge was 13.3 cm/year in the AP1 and 6.70 cm/year in the AP2, while in the pond with aeration this was on the average of 0.5 cm/year, due to the aeration regime. The sludge can be used as a fertilizer in agriculture, if the chemical characteristics of the soil are taken into account so as to avoid the accumulation of nutrients and damage to plants. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
15. Reduction of odors from a facultative pond using two different operating practices.
- Author
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Truppel, A., Camargos, J. L. M., da Costa, R. H. R., and Filho, P. Belli
- Subjects
SEWAGE purification ,ODOR control ,ANALYTICAL chemistry ,SEWAGE disposal plants ,DEODORIZATION ,OLFACTOMETRY - Abstract
This paper presents the results of a proposed intervention to deal with the odor problems of a sewage treatment works (STW), which is located near a populated area. The STW consists of a facultative pond. Since this pond functions under close to anaerobic conditions, unpleasant odors are emitted. In this respect, two possible ways to deodorize the pond were evaluated. Firstly, the recirculation of effluent using 1/6 of the flow stream followed by aeration of the pond with a reduced power aerator. In order to study the efficiencies of the deodorization methodologies chemical analyses of the gases NH
3 and H2 S, olfactometric analyses and evaluation of the environmental perception of the population in relation to the odors originating from the STW, were carried out for each experimental situation. The results showed a significant reduction in odors when aeration with reduced power equipment was utilized in combination with recirculation of effluent in the pond. Reductions in emissions of H2 S from 0.1345 mg/m³ to 0.0083 mg/m³ and of NH3 from 0.021 mg/m³ to 0.0073 mg/m³ were obtained. To analyze the behavior of the pond, its planktonic community was investigated, with a difference in species for the situations with and without odor being observed. [ABSTRACT FROM AUTHOR]- Published
- 2005
- Full Text
- View/download PDF
16. Optimization of the treatment of piggery wastes in water hyacinth ponds.
- Author
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Costa, R. H. R., Zanotelli, C. T., Hoffmann, D. M., Belli Filho, P., Perdomo, C. C., and Rafikov, M.
- Subjects
- *
PLANT spacing , *WATER hyacinth , *ANIMAL waste , *SLAUGHTERHOUSE by-products , *SEWAGE lagoons , *SURFACE impoundments , *BIOLOGICAL nutrient removal , *BIOREMEDIATION , *MATHEMATICAL models , *DENITRIFICATION , *NITROGEN removal (Sewage purification) - Abstract
This work investigates the optimal management of water hyacinth ponds for the improvement of piggery waste treatment. The optimal harvesting strategy for the water hyacinth was studied using a single mathematical model. The water hyacinth optimal harvesting problem was formulated as an optimal control problem that was solved by application of Pontryagin's Maximum Principle. The optimization of the water hyacinth control in the pond indicates that the plant density should be reduced whenever it reaches half of the maximum capacity for growth. Two experimental systems were used to validate the mathematical model, one in real scale and the other in pilot scale. The results demonstrated the feasibility of the proposed harvesting strategy. For example, a comparison of the total nitrogen removal in the different pilot ponds confirmed the modeling results, in that the performance of the pond maintained with 50% water hyacinth cover was better than the others. [ABSTRACT FROM AUTHOR]
- Published
- 2003
- Full Text
- View/download PDF
17. Biogas production, sludge accumulation and mass balance of carbon in anaerobic ponds.
- Author
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Picot, B., Paing, J., Sambuco, J. P., Costa, R. H. R., and Rambaud, A.
- Subjects
BIOGAS ,CARBON ,SEWAGE lagoons ,ANAEROBIC digestion ,SEWAGE sludge digestion ,BIOLOGICAL nutrient removal ,METHANE - Abstract
This work concerned the application of anaerobic ponds for the primary treatment of urban wastewater in a Mediterranean climate. It was carried out on anaerobic ponds at large scale in Mèze (France). The anaerobic ponds constitute a good primary treatment with the removal of 55% of SS and 30% of BOD
5 , with a small surface area. The accumulation rate of sludge was only 0.017 m³/capita.year, due to their intensive anaerobic degradation. The anaerobic digestion reached equilibrium after one year of operation. The accumulation of sludge then showed seasonal variations with a substantial accumulation in winter and the digestion of the stock in summer. This change can be related to the influence of the temperature on methanogenesis. The production of biogas (83% CH4 ) was measured by gas collectors especially developed for this study and was also strongly dependent on temperature. The mass balance of carbon showed that 74% of the removed organic carbon was converted into CH4 , 13% into dissolved inorganic carbon and 15% was stored in sludge. However, the anaerobic ponds presented a risk of creating odor nuisances with the emission of H2 S. [ABSTRACT FROM AUTHOR]- Published
- 2003
- Full Text
- View/download PDF
18. Influence of load distribution and recycle rate in step-fed facultative ponds .
- Author
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Sambuco, J. P., Da Costa, R. H. R., Paing, J., and Picot, B.
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SEWAGE purification , *WATER utilities , *EFFLUENT quality , *LAND treatment of wastewater , *SEWAGE disposal plants , *WATER reuse - Abstract
Presents the results of research on a wastewater treatment system with four identical facultative ponds in series with step-feeding and recirculation (SFFPR). Research of four modes of distribution of the influent; How the four ponds were homogeneous; Findings that the ponds receiving the highest loading showed a bacterial biomass higher than that of the primary production.
- Published
- 2002
- Full Text
- View/download PDF
19. Performance of a baffled facultative pond treating piggery wastes .
- Author
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Zanotelli, C. T., Medri, W., Belli Filho, P., Perdomo, C., Mulinari, M. R., and Costa, R. H. R.
- Subjects
SEWAGE lagoons ,SEWAGE disposal plants ,WASTE management ,WASTE products ,SEWERAGE ,ORGANIC wastes ,OXIDATION ditches ,SURFACE impoundments - Abstract
Presents a study to evaluate the performance of a baffled facultative pond for the treatment of piggery wastes. Use of an equalizer, one decanter, two anaerobic ponds, one facultative pond, and one maturation pond with water hyacinths; How research was conducted in the west region of Santa Catarina, Brazil; Findings that the first baffle in the facultative pond was mainly responsible for the efficiency of this pond.
- Published
- 2002
- Full Text
- View/download PDF
20. High-rate pond for treatment of piggery wastes
- Author
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Medri, W., Costa, R. H. R., and Perdomo, C. C.
- Subjects
- *
LIVESTOCK , *SEWAGE , *SEWAGE purification , *HYDROLOGY - Abstract
This work deals with studies on high-rate ponds, a batch working system that is followed by a filter pond where Chinese carp were introduced for piggery wastes treatment. COD removal values for the high-rate pond were in the order of 95% in the summer and 70% in the winter for an initial concentration of 2000 mg/L. Total nitrogen removal values ranged between 90% and 60%, respectively, in summer and winter periods for an initial concentration of 600 mg/L. Seasonal variations, which are mainly observed under differences of temperature, were shownto be not relevant for total phosphorus removal, a process that appears to depend mainly on increases of pH values over 8.5. An hydraulicretention time ranging between 15 to 20 days was found to be best for pond functioning. The plug flow model fits well to the pond's physical characteristics. The filter pond was shown to be a great potential process for removal of algae produced in the high-rate pond. [ABSTRACT FROM AUTHOR]
- Published
- 2000
21. Tertiary treatment of piggery wastes in water hyacinth ponds
- Author
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Bavaresco, A. S. L., Philippi, L. S., Medri, W., and Costa, R. H. R.
- Subjects
SEWAGE purification ,SEWAGE ,HYDROLOGY ,LIVESTOCK - Abstract
This work was developed in two stages: initially in water hyacinth ponds under batch conditions (pilot-scale) and later in full-scale ponds under a continuous state, so as to determine local parameters for the operation of this kind of pond for piggery wastes treatment. Seasonal variations are important features influencing plant productivityand performance. The water hyacinth ponds were able to remove around50% of the applied organic loads (COD, BOD, TN, TP), even in case ofelevated values (110 kg/ha/day) applied for total nitrogen surface loads. An hydraulic retention time of 20 days was shown to be ideal for the treatment. The utilization of water hyacinths as a complement for the animal diet closes the productive cycle in a sustainable way. The results have demonstrated the feasibility of the proposed treatment system. [ABSTRACT FROM AUTHOR]
- Published
- 2000
22. The effects of a full-scale anaerobic side-stream reactor on sludge decay and biomass activity.
- Author
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Velho VF, Andreottola G, Foladori P, and Costa RHR
- Subjects
- Anaerobiosis, Biomass, Sewage, Bioreactors, Waste Disposal, Fluid methods
- Abstract
A full-scale anaerobic side-stream reactor (ASSR) for sludge reduction was monitored in terms of sludge production and compared with the previous conventional activated sludge configuration (CAS). A detailed solid mass balance was calculated on the whole full-scale plant to estimate the sludge reduction associated with the ASSR. The activity of the biomass, which undergoes alternation of aerobic and anaerobic conditions, was investigated by the respirometric test. The ASSR promoted a reduction of heterotrophic biomass activity and the substrate consumption rate in the activated sludge implemented with ASSR (AS + ASSR) was 36% smaller than in the CAS period. The solid mass balance indicated a sludge reduction of 28%. During the 270-day operation, the observed sludge yield passed from 0.438 kgTSS/kgCOD in the CAS to 0.315 in the AS + ASSR configuration. The solubilization of chemical oxygen demand (COD), NH
4 + -N and orthophosphate were verified under anaerobic conditions. The results suggest that the possible mechanisms of sludge reduction were the increase of the system sludge retention time (SRT) by ASSR addition, and the reduction in heterotrophic biomass activity added to the organic compounds' hydrolysis.- Published
- 2019
- Full Text
- View/download PDF
23. Effect of COD/N ratio on N 2 O production during nitrogen removal by aerobic granular sludge.
- Author
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Velho VF, Magnus BS, Daudt GC, Xavier JA, Guimarães LB, and Costa RHR
- Subjects
- Aerobiosis, Ammonia, Biological Oxygen Demand Analysis, Denitrification, Heterotrophic Processes, Nitrification, Nitrites, Nitrogen metabolism, Nitrous Oxide analysis, Waste Disposal, Fluid methods, Bioreactors microbiology, Nitrogen chemistry, Nitrous Oxide metabolism, Sewage microbiology
- Abstract
N
2 O-production was investigated during nitrogen removal using aerobic granular sludge (AGS) technology. A pilot sequencing batch reactor (SBR) with AGS achieved an effluent in accordance with national discharge limits, although presented a nitrite accumulation rate of 95.79% with no simultaneous nitrification-denitrification. N2 O production was 2.06 mg L-1 during the anoxic phase, with N2 O emission during air pulses and the aeration phase of 1.6% of the nitrogen loading rate. Batch tests with AGS from the pilot reactor verified that at the greatest COD/N ratio (1.55), the N2 O production (1.08 mgN2 O-N L-1 ) and consumption (up to 0.05 mgN2 O-N L-1 ), resulted in the lowest remaining dissolved N2 O (0.03 mgN2 O-N L-1 ), stripping the minimum N2 O gas (0.018 mgN2 O-N L-1 ). Conversely, the carbon supply shortage, under low C/N ratios, increased N2 O emission (0.040 mgN2 O-N L-1 ), due to incomplete denitrification. High abundance of ammonia-oxidizing and low abundance of nitrite-oxidizing bacteria were found, corroborating the fact of partial nitrification. A denitrifying heterotrophic community, represented mainly by Pseudoxanthomonas, was predominant in the AGS. Overall, the AGS showed stable partial nitrification ability representing capital and operating cost savings. The SBR operation flexibility could be advantageous for controlling N2 O emissions, and extending the anoxic phase would benefit complete denitrification in cases of low C/N influents.- Published
- 2017
- Full Text
- View/download PDF
24. Anaerobic side-stream reactor for excess sludge reduction: 5-year management of a full-scale plant.
- Author
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Velho VF, Foladori P, Andreottola G, and Costa RH
- Subjects
- Aerobiosis, Anaerobiosis, Bioreactors, Sewage, Waste Management methods, Wastewater
- Abstract
The long-term performances of a full-scale anaerobic side-stream reactor (ASSR) aimed at sludge reduction have been monitored for the first time, in comparison with a conventional activated sludge process (CAS). The plant was integrated with an ASSR treatment of 2293-3293 m(3). Operational parameters in the ASSR were: ORP -250 mV, interchange ratio of 7-10%, hydraulic retention time of 7 d. No worsening of effluent quality was observed in the ASSR configuration and removal efficiency of COD and NH4 was above 95%. A slight increase in the Sludge Volume Index did not cause worsening in effluent solids concentration. The observed sludge yield (Yobs) passed from 0.44 kgTSS/kgCOD in the CAS to 0.35 in the ASSR configuration. The reduction of Yobs by 20% is lower than expected from the literature where sythetic wastewater is used, indicating that sludge reduction efficiency is largely affected by inert mass fed with influent real wastewater. An increase by 45% of the ASSR volume did not promote a further reduction of Yobs, because sludge reduction is affected not solely by endogenous decay but also by other factors such as interchange ratio and aerobiosis/anaerobiosis alternation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
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25. Concerning the role of cell lysis-cryptic growth in anaerobic side-stream reactors: the single-cell analysis of viable, dead and lysed bacteria.
- Author
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Foladori P, Velho VF, Costa RH, Bruni L, Quaranta A, and Andreottola G
- Subjects
- Aerobiosis, Anaerobiosis, Bacteria enzymology, Biological Oxygen Demand Analysis, Nitrogen metabolism, Oxidation-Reduction, Sewage chemistry, Single-Cell Analysis, Bacteria growth & development, Bioreactors microbiology, Sewage microbiology, Waste Disposal, Fluid methods
- Abstract
In the Anaerobic Side-Stream Reactor (ASSR), part of the return sludge undergoes alternating aerobic and anaerobic conditions with the aim of reducing sludge production. In this paper, viability, enzymatic activity, death and lysis of bacterial cells exposed to aerobic and anaerobic conditions for 16 d were investigated at single-cell level by flow cytometry, with the objective of contributing to the understanding of the mechanisms of sludge reduction in the ASSR systems. Results indicated that total and viable bacteria did not decrease during the anaerobic phase, indicating that anaerobiosis at ambient temperature does not produce a significant cell lysis. Bacteria decay and lysis occurred principally under aerobic conditions. The aerobic decay rate of total bacteria (bTB) was considered as the rate of generation of lysed bacteria. Values of bTB of 0.07-0.11 d(-1) were measured in anaerobic + aerobic sequence. The enzymatic activity was not particularly affected by the transition from anaerobiosis to aerobiosis. Large solubilisation of COD and NH4(+) was observed only under anaerobic conditions, as a consequence of hydrolysis of organic matter, but not due to cell lysis. The observations supported the proposal of two independent mechanisms contributing equally to sludge reduction: (1) under anaerobic conditions: sludge hydrolysis of non-bacterial material, (2) under aerobic conditions: bacterial cell lysis and oxidation of released biodegradable compounds., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2015
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26. Micronucleus induction in mussels exposed to okadaic acid.
- Author
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Carvalho Pinto-Silva CR, Ferreira JF, Costa RH, Belli Filho P, Creppy EE, and Matias WG
- Subjects
- Animals, Environmental Monitoring methods, Micronucleus Tests, Time Factors, Bivalvia drug effects, Bivalvia genetics, Hemocytes drug effects, Marine Toxins toxicity, Okadaic Acid toxicity
- Abstract
Some toxins present in the marine environment are capable of inducing mutagenicity and/or carcinogenicity. Among these toxins, okadaic acid (OA) is gaining considerable interest since it induces DNA based modifications at low concentrations and accumulates in filter-feeding marine animals, including those used for human consumption. This study aims to evaluate the genotoxicity of OA in the haemocytes of the mussel Perna perna, using the micronucleus assay. Fifty-four mussels were separated into three groups of 18 animals. One group received 0.3 microg of OA diluted in 10 microl of ethanol and ultrapure water while the other groups were considered as controls and were exposed to a solvent plus seawater mixture. A significantly higher frequency of micronuclei was observed in haemocytes from the OA-exposed group. There were no statistical differences between the two control groups.
- Published
- 2003
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27. Human peroxisome proliferator-activated receptor alpha (PPARalpha) supports the induction of peroxisome proliferation in PPARalpha-deficient mouse liver.
- Author
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Yu S, Cao WQ, Kashireddy P, Meyer K, Jia Y, Hughes DE, Tan Y, Feng J, Yeldandi AV, Rao MS, Costa RH, Gonzalez FJ, and Reddy JK
- Subjects
- Animals, Cell Division drug effects, Gene Expression Regulation, Humans, Mice, Mice, Inbred C57BL, Mice, Knockout, Oxidation-Reduction, Peroxisomes drug effects, Pyrimidines pharmacology, RNA, Messenger analysis, Liver metabolism, Peroxisomes metabolism, Receptors, Cytoplasmic and Nuclear physiology, Transcription Factors physiology
- Abstract
Peroxisome proliferators, which function as peroxisome proliferator-activated receptor alpha (PPARalpha) agonists, induce peroxisomal, microsomal, and mitochondrial fatty acid oxidation enzymes, in conjunction with peroxisome proliferation, in liver cells. Sustained activation of PPARalpha leads to the development of liver tumors in rats and mice. The assertion that synthetic PPARalpha ligands pose negligible carcinogenic risk to humans is attributable, in part, to the failure to observe peroxisome proliferation in human hepatocytes. To explore the mechanism(s) of species-specific differences in response to PPARalpha ligands, we determined the functional competency of human PPARalpha in vivo and compared its potency with that of mouse PPARalpha. Recombinant adenovirus that expresses human or mouse PPARalpha was produced and administered intravenously to PPARalpha-deficient mice. Human as well as mouse PPARalpha fully restored the development of peroxisome proliferator-induced immediate pleiotropic responses, including peroxisome proliferation and enhanced expression of genes involved in lipid metabolism as well as nonperoxisomal genes, such as CD36, Ly-6D, Rbp7, monoglyceride lipase, pyruvate dehydrogenase kinase-4, and C3f, that have been identified recently to be up-regulated in livers with peroxisome proliferation. These studies establish that human PPARalpha is functionally competent and is equally as dose-sensitive as mouse PPARalpha in inducing peroxisome proliferation within the context of mouse liver environment and that it can heterodimerize with mouse retinoid X receptor, and this human PPARalpha-mouse retinoid X receptor chimeric heterodimer transcriptionally activates mouse PPARalpha target genes in a manner qualitatively similar to that of mouse PPARalpha.
- Published
- 2001
- Full Text
- View/download PDF
28. DDB2 induces nuclear accumulation of the hepatitis B virus X protein independently of binding to DDB1.
- Author
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Nag A, Datta A, Yoo K, Bhattacharyya D, Chakrabortty A, Wang X, Slagle BL, Costa RH, and Raychaudhuri P
- Subjects
- G1 Phase, Hepatocytes metabolism, Humans, Protein Subunits, Tumor Cells, Cultured, Viral Regulatory and Accessory Proteins, Cell Nucleus metabolism, DNA-Binding Proteins metabolism, DNA-Binding Proteins physiology, Trans-Activators metabolism
- Abstract
The hepatitis B virus (HBV) X protein (HBx) is critical for the life cycle of the virus. HBx associates with several host cell proteins including the DDB1 subunit of the damaged-DNA binding protein DDB. Recent studies on the X protein encoded by the woodchuck hepadnavirus have provided correlative evidence indicating that the interaction with DDB1 is important for establishment of infection by the virus. In addition, the interaction with DDB1 has been implicated in the nuclear localization of HBx. Because the DDB2 subunit of DDB is required for the nuclear accumulation of DDB1, we investigated the role of DDB2 in the nuclear accumulation of HBx. Here we show that expression of DDB2 increases the nuclear levels of HBx. Several C-terminal deletion mutants of DDB2 that fail to bind DDB1 are able to associate with HBx, suggesting that DDB2 may associate with HBx independently of binding to DDB1. We also show that DDB2 enhances the nuclear accumulation of HBx independently of binding to DDB1, since a mutant that does not bind DDB1 is able to enhance the nuclear accumulation of HBx. HBV infection is associated with liver pathogenesis. We show that the nuclear levels of DDB1 and DDB2 are tightly regulated in hepatocytes. Studies with regenerating mouse liver indicate that during late G1 phase the nuclear levels of both subunits of DDB are transiently increased, followed by a sharp decrease in S phase. Taken together, these results suggest that DDB1 and DDB2 would participate in the nuclear functions of HBx effectively only during the late-G1 phase of the cell cycle.
- Published
- 2001
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29. Increased levels of forkhead box M1B transcription factor in transgenic mouse hepatocytes prevent age-related proliferation defects in regenerating liver.
- Author
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Wang X, Quail E, Hung NJ, Tan Y, Ye H, and Costa RH
- Subjects
- Aging genetics, Animals, Cell Division, Cyclin-Dependent Kinases metabolism, Cyclophilins genetics, DNA Replication, Forkhead Box Protein M1, Forkhead Transcription Factors, Hepatocytes physiology, Mice, Mice, Transgenic, Phosphorylation, RNA Probes, RNA, Antisense, Retinoblastoma Protein metabolism, Ribonucleases, S Phase, Transfection, Aging physiology, DNA-Binding Proteins genetics, Hepatocytes cytology, Liver physiology, Liver Regeneration physiology, Transcription Factors genetics
- Abstract
The forkhead box (Fox) family of transcription factors share homology in the winged helix/forkhead DNA-binding domain and play important roles in regulating cellular proliferation, differentiation, longevity, and cellular transformation. Forkhead box M1B (FoxM1B) is a ubiquitously expressed member of the Fox transcription factor family whose expression is restricted to proliferating cells and that mediates hepatocyte entry into DNA synthesis and mitosis during liver regeneration. Recent cDNA microarray studies indicated that age-related defects in cellular proliferation are associated with diminished expression of the FoxM1B transcription factor. Here, we show that increased levels of FoxM1B in regenerating liver of old transgenic mice restore the sharp peaks in hepatocyte DNA replication and mitosis that are the hallmarks of young regenerating mouse liver. Restoration of the young regenerating liver phenotype is associated with increased expression of numerous cell cycle regulatory genes that include cyclin D1, cyclin A2, cyclin F, cyclin B1, cyclin B2, Cdc25B, and p55cdc. Cotransfection assays in the human hepatoma HepG2 cell line demonstrated that FoxM1B protein stimulated expression of both the cyclin B1 and cyclin D1 promoters, suggesting that these cyclin genes are a direct FoxM1B transcriptional target. These results suggest that FoxM1B controls the transcriptional network of genes that are essential for cell division and exit from mitosis. Our results indicate that reduced expression of the FoxM1B transcription factor contributes to the decline in cellular proliferation observed in the aging process.
- Published
- 2001
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30. Defects in pulmonary vasculature and perinatal lung hemorrhage in mice heterozygous null for the Forkhead Box f1 transcription factor.
- Author
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Kalinichenko VV, Lim L, Stolz DB, Shin B, Rausa FM, Clark J, Whitsett JA, Watkins SC, and Costa RH
- Subjects
- Alleles, Animals, Apoptosis, Blotting, Western, Bone Morphogenetic Protein 4, Bone Morphogenetic Proteins metabolism, DNA, Complementary metabolism, Dose-Response Relationship, Drug, Endothelial Growth Factors metabolism, Endothelium metabolism, Forkhead Transcription Factors, Hemorrhage, Heterozygote, Immunohistochemistry, In Situ Nick-End Labeling, Lung pathology, Lymphokines metabolism, Mice, Mice, Knockout, Microscopy, Electron, Models, Genetic, Mutation, Oligonucleotide Array Sequence Analysis, Phenotype, Platelet Endothelial Cell Adhesion Molecule-1 metabolism, RNA, Messenger metabolism, Receptor Protein-Tyrosine Kinases metabolism, Receptors, Growth Factor metabolism, Receptors, Vascular Endothelial Growth Factor, Trans-Activators metabolism, Transcription Factors physiology, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factors, beta-Galactosidase metabolism, DNA-Binding Proteins, Lung embryology, Lung metabolism, Transcription Factors biosynthesis, Transcription Factors genetics, Transcription Factors metabolism
- Abstract
Decreased pulmonary expression of Forkhead Box f1 (Foxf1) transcription factor was associated with lethal alveolar hemorrhage in 55% of the Foxf1 +/- newborn mice. The severity of the pulmonary abnormalities correlates with the levels of Foxf1 mRNA. Defects in alveolarization and vasculogenesis were observed in subsets of the Foxf1 +/- mice with relatively low levels of expression from the normal Foxf1 allele. Lung hemorrhage was coincident with disruption of the mesenchymal-epithelial cell interfaces in the alveolar and bronchiolar regions of the lung parenchyma and was associated with increased apoptosis and reduced surfactant protein B (SP-B) expression. Finally, the lung defect associated with the Foxf1 +/- mutation was accompanied by reduced expression of vascular endothelial growth factor (VEGF), the VEGF receptor 2 (Flk-1), bone morphogenetic protein 4 (Bmp-4), and the transcription factors of the Brachyury T-Box family (Tbx2-Tbx5) and Lung Kruppel-like Factor. Reduction in the level of Foxf1 caused neonatal pulmonary hemorrhage and abnormalities in alveologenesis, implicating this transcription factor in the regulation of mesenchyme-epithelial interaction critical for lung morphogenesis., (Copyright 2001 Academic Press.)
- Published
- 2001
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31. Earlier expression of the transcription factor HFH-11B diminishes induction of p21(CIP1/WAF1) levels and accelerates mouse hepatocyte entry into S-phase following carbon tetrachloride liver injury.
- Author
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Wang X, Hung NJ, and Costa RH
- Subjects
- Animals, Carbon Tetrachloride, Cell Adhesion Molecules metabolism, Cell Cycle Proteins metabolism, Chemical and Drug Induced Liver Injury, Cyclin-Dependent Kinase Inhibitor p21, Cyclins metabolism, Forkhead Box Protein M1, Forkhead Transcription Factors, Liver Diseases physiopathology, Male, Mice, Mice, Inbred Strains, Mice, Transgenic genetics, Time Factors, Transcription Factors metabolism, cdc25 Phosphatases metabolism, Cyclins genetics, Gene Expression physiology, Gene Expression Regulation physiology, Hepatocytes physiology, Liver Diseases genetics, Liver Diseases pathology, S Phase, Transcription Factors genetics
- Abstract
Partial hepatectomy (PH) or toxic liver injury induces the proliferation of terminally differentiated hepatic cells to regenerate the original size of the adult liver. Previous PH liver regeneration studies showed that premature transgenic expression of the Forkhead Box M1b (FoxM1b, HFH-11B) transcription factor accelerated hepatocyte entry into DNA replication (S-phase). In this study, we used carbon tetrachloride (CCl(4)) liver injury to induce a different type of mouse liver regeneration and show that premature hepatic HFH-11B levels also accelerate the onset of hepatocyte S-phase in this injury model. Unlike PH liver regeneration, earlier hepatocyte proliferation after CCl(4) liver injury is correlated with diminished transgenic hepatic levels of p21(CIP1/WAF1) at the G1/S transition of the cell cycle. Differential hybridization of cDNA arrays and RNase protection studies determined that CCl(4) regenerating liver of transgenic mice displayed early stimulated expression of the S-phase promoting cyclin D1 and cyclin E and sustained levels of Cdc25a phosphatase genes. Compared with previous PH liver regeneration studies, our data suggest that premature expression of HFH-11B activates distinct S-phase promotion pathways in the CCl(4) liver injury model. Although proliferating transgenic hepatocytes induced by either PH or CCl(4) liver injury displayed early expression of identical M-phase cyclin genes (cyclin B1, B2, A2, and F), only CCl(4) regenerating transgenic liver exhibited earlier expression of the M-phase promoting Cdc25b. These studies suggest that CCl(4) injury of transgenic liver not only uses the same mechanisms as PH to mediate accelerated hepatocyte entry into mitosis, but also promotes M-phase entry by stimulating Cdc25b expression.
- Published
- 2001
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32. Transcription factors in mouse lung development and function.
- Author
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Costa RH, Kalinichenko VV, and Lim L
- Subjects
- Amino Acid Motifs, Animals, Cell Differentiation, Epithelial Cells cytology, Epithelial Cells metabolism, Gene Expression Regulation, Developmental, Lung cytology, Mesoderm cytology, Mesoderm metabolism, Mice, Mice, Knockout, Mice, Transgenic, Morphogenesis physiology, Phenotype, Transcription Factors genetics, Lung embryology, Lung metabolism, Transcription Factors metabolism
- Abstract
Development of the mouse lung initiates on day 9.5 postcoitum from the laryngotracheal groove and involves mesenchymal-epithelial interactions, in particular, those between the splanchnic mesoderm and epithelial cells (derived from foregut endoderm) that induce cellular proliferation, migration, and differentiation, resulting in branching morphogenesis. This developmental process mediates formation of the pulmonary bronchiole tree and integrates a terminal alveolar region with an extensive endothelial capillary bed, which facilitates efficient gas exchange with the circulatory system. The major function of the mesenchymal-epithelial signaling is to potentiate the activity or expression of cell type-specific transcription factors in the developing lung, which, in turn, cooperatively bind to distinct promoter regions and activate target gene expression. In this review, we focus on the role of transcription factors in lung morphogenesis and the maintenance of differentiated gene expression. These lung transcription factors include forkhead box A2 [also known as hepatocyte nuclear factor (HNF)-3beta], HNF-3/forkhead homolog (HFH)-8 [also known as FoxF1 or forkhead-related activator-1], HNF-3/forkhead homolog-4 (also known as FoxJ1), thyroid transcription factor-1 (Nkx2.1), and homeodomain box A5 transcription factors, the zinc finger Gli (mouse homologs of the Drosophila cubitus interruptus) and GATA transcription factors, and the basic helix-loop-helix Pod1 transcription factor. We summarize the phenotypes of transgenic and knockout mouse models, which define important functions of these transcription factors in cellular differentiation and lung branching morphogenesis.
- Published
- 2001
- Full Text
- View/download PDF
33. Differential expression of forkhead box transcription factors following butylated hydroxytoluene lung injury.
- Author
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Kalinichenko VV, Lim L, Shin B, and Costa RH
- Subjects
- Animals, Bronchi metabolism, Bronchi pathology, Butylated Hydroxytoluene, Cell Division physiology, Endothelium metabolism, Endothelium pathology, Forkhead Box Protein M1, Forkhead Transcription Factors, Hepatocyte Nuclear Factor 3-beta, Lung metabolism, Lung pathology, Lung Diseases chemically induced, Lung Diseases pathology, Male, Mesoderm pathology, Mice, Mice, Inbred BALB C, Muscle, Smooth cytology, Muscle, Smooth metabolism, DNA-Binding Proteins metabolism, Lung Diseases metabolism, Nuclear Proteins metabolism, Trans-Activators metabolism, Transcription Factors metabolism
- Abstract
The forkhead box (Fox) proteins are a growing family of transcription factors that have important roles in cellular proliferation and differentiation and in organ morphogenesis. The Fox family members hepatocyte nuclear factor (HNF)-3beta (Foxa2) and HNF-3/forkhead homolog (HFH)-8 (FREAC-1, Foxf1) are expressed in adult pulmonary epithelial and mesenchymal cells, respectively, but these cells display only low expression levels of the proliferation-specific HFH-11B gene (Trident, Foxm1b). The regulation of these Fox transcription factors in response to acute lung injury, however, has yet to be determined. We report here on the use of butylated hydroxytoluene (BHT)-mediated lung injury to demonstrate that HFH-11 protein and RNA levels were markedly increased throughout the period of lung repair. The maximum levels of HFH-11 were observed by day 2 following BHT injury when both bronchiolar and alveolar epithelial cells were undergoing extensive proliferation. Although BHT lung injury did not alter epithelial cell expression of HNF-3beta, a 65% reduction in HFH-8 mRNA levels was observed during the period of mesenchymal cell proliferation. HFH-8-expressing cells were colocalized with platelet endothelial cell adhesion molecule-1-positive alveolar endothelial cells and with alpha-smooth muscle actin-positive peribronchiolar smooth muscle cells.
- Published
- 2001
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34. Atypical mouse cerebellar development is caused by ectopic expression of the forkhead box transcription factor HNF-3beta.
- Author
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Zhou H, Hughes DE, Major ML, Yoo K, Pesold C, and Costa RH
- Subjects
- Animals, Animals, Newborn, Antigens, CD genetics, Apoptosis, Astrocytes cytology, Astrocytes metabolism, Basic Helix-Loop-Helix Transcription Factors, Cell Adhesion Molecules, Neuronal genetics, Cell Differentiation, Cell Movement, Cerebellum abnormalities, Cerebellum cytology, DNA-Binding Proteins genetics, DNA-Binding Proteins immunology, ErbB Receptors genetics, Extracellular Matrix Proteins genetics, Gene Expression, Genetic Markers genetics, Hepatocyte Nuclear Factor 3-beta, In Situ Hybridization, Insulin-Like Growth Factor Binding Protein 1 genetics, Insulin-Like Growth Factor Binding Protein 1 metabolism, Integrin alpha5, Mice, Mice, Transgenic, Nerve Tissue Proteins, Netrin Receptors, Neuroglia cytology, Neuroglia metabolism, Neurons cytology, Neurons metabolism, Nuclear Proteins genetics, Nuclear Proteins immunology, Oligonucleotide Array Sequence Analysis, Phenotype, Purkinje Cells cytology, Purkinje Cells metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Receptor, ErbB-4, Receptors, Nerve Growth Factor genetics, Reelin Protein, Serine Endopeptidases, Transcription Factors genetics, Cerebellum growth & development, Cerebellum metabolism, DNA-Binding Proteins metabolism, Nuclear Proteins metabolism
- Abstract
To assess the role of hepatocyte nuclear factor-3beta (HNF-3beta) in hepatocyte-specific gene transcription, we reported the characterization of the liver phenotype with transgenic mice in which the -3-kb transthyretin (TTR) promoter functioned to increase HNF-3beta expression. During breeding of the TTR-HNF-3beta transgenic mice we noticed that they displayed severe ataxia. In this study, we describe the analysis of our transgenic cerebellar phenotype and demonstrate that ectopic expression of HNF-3beta disrupted cerebellar morphogenesis and caused reduction in cerebellar size. In postnatal cerebellum, the HNF-3beta transgene expression pattern is colocalized to glial fibrillary acidic protein-positive cerebellar astrocytes and Bergmann glial cells. As a result of protracted expression, the transgenic cerebella are impaired in terms of astrocyte dispersal and formation of Bergmann glial cell processes. This caused a disruption in neuronal cell migration to the cortical laminar layers and Purkinje dendritic arbor maturation, thus leading to diminished foliation. Differential hybridization of cDNA arrays was used to identify altered expression of cerebellar genes, which is consistent with the observed defect in transgenic cerebellar morphogenesis and size as well as glial maturation. These include diminished expression of the brain lipid-binding protein, which is required for glial morphological differentiation, and the basic helix-loop-helix NeuroD/Beta2 and homeodomain Engrailed-2 transcription factors, which are required for normal cerebellar morphogenesis and foliation. Undetectable levels of ataxia telangiectasia (ATM), which is required for proper development of the Purkinje dendritic arbor, were found in postnatal transgenic cerebella. Furthermore, the transgenic cerebella displayed levels of insulin-like growth factor binding protein-1 elevated to 22 times greater than those measured for wild-type cerebella, an elevation consistent with the reduction in transgenic cerebellar size.
- Published
- 2001
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35. Adenovirus-mediated increase of HNF-3 levels stimulates expression of transthyretin and sonic hedgehog, which is associated with F9 cell differentiation toward the visceral endoderm lineage.
- Author
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Tan Y, Costa RH, Kovesdi I, and Reichel RR
- Subjects
- Animals, Cell Line, Cell Lineage, Chloramphenicol O-Acetyltransferase genetics, DNA-Binding Proteins metabolism, Endoderm, Genetic Vectors, Hedgehog Proteins, Hepatocyte Nuclear Factor 3-alpha, Hepatocyte Nuclear Factor 3-beta, Humans, Mice, Nuclear Proteins metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Tumor Cells, Cultured, Adenoviridae genetics, Cell Differentiation genetics, DNA-Binding Proteins biosynthesis, Gene Expression Regulation genetics, Nuclear Proteins biosynthesis, Prealbumin genetics, Trans-Activators genetics, Transcription Factors
- Abstract
Retinoic acid-induced differentiation of mouse F9 embryonal carcinoma cells toward the visceral endoderm lineage is accompanied by increased expression of the Forkhead Box (Fox) transcription factors hepatocyte nuclear factor 3a (HNF-3alpha) and HNF-3beta, suggesting that they play a crucial role in visceral endoderm development. Retinoic acid stimulation results in a cascade of HNF-3 induction in which HNF-3alpha is a primary target for retinoic acid action and its increase is required for subsequent induction of HNF-3beta expression. Increased expression of HNF-3beta precedes activation of its known target genes, including transthyretin (TTR), Sonic hedgehog (Shh), HNF-1alpha, HNF-1beta, and HNF-4alpha. In order to examine whether increased HNF-3 expression is sufficient to induce expression of its downstream target genes without retinoic acid stimulation, we have used adenovirus-based expression vectors to increase HNF-3 protein levels in F9 cells. We demonstrate that adenovirus-mediated increase of HNF-3alpha levels in F9 cells is sufficient to induce activation of endogenous HNF-3beta levels followed by increased TTR and Shh expression. Furthermore, we show that elevated HNF-3beta levels stimulate expression of endogenous TTR and Shh without retinoic acid stimulation. Moreover, ectopic HNF-3 levels in undifferentiated F9 cells are insufficient to induce HNF-3alpha, HNF-1alpha, HNF-1beta, and HNF-4alpha expression, suggesting that their transcriptional activation required other regulatory proteins induced by the retinoic acid differentiation program. Finally, our studies demonstrate the utility of cell infections with adenovirus expressing distinct transcription factors to identify endogenous target genes, which are assembled with the appropriate nucleosome structure.
- Published
- 2001
- Full Text
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36. Odor control of an anaerobic lagoon with a biological cover: floating peat beds.
- Author
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Picot B, Paing J, Toffoletto L, Sambuco JP, and Costa RH
- Subjects
- Air Pollution prevention & control, Bacteria, Anaerobic physiology, Biodegradation, Environmental, Oxidation-Reduction, Plants, Sulfur chemistry, Waste Disposal, Fluid, Hydrogen Sulfide analysis, Odorants, Soil
- Abstract
The use of a biological cover for in situ control of gaseous sulfide emission from an anaerobic pond was investigated by a laboratory-scale experiment. The biological cover, constituting by a peat bed floating on the wastewater, caused a reduction of the H2S emission rate by 84.6%. The addition of Fe3+ (with FeCl3) and plants (Juncus effusus L.) to the peat bed significantly improved the performance to reach a H2S removal of 95.5%. Despite the fluctuations in the sulfide concentration in the wastewater, the performance of the biological covers remained constant during the entire period of the study. The analysis of the different forms of sulfur accumulated in the peat beds allowed the understanding of the mechanisms involved in H2S removal. The high amount of sulfate demonstrated that the conditions were favorable to the biological oxidation of H2S. The addition of Fe3+ increased the formation of insoluble ferrous monosulfide (FeS) and pyrite (FeS2). The plants seemed to convert sulfate into elemental and organic sulfur.
- Published
- 2001
37. Elevated levels of hepatocyte nuclear factor 3beta in mouse hepatocytes influence expression of genes involved in bile acid and glucose homeostasis.
- Author
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Rausa FM, Tan Y, Zhou H, Yoo KW, Stolz DB, Watkins SC, Franks RR, Unterman TG, and Costa RH
- Subjects
- ATP Binding Cassette Transporter, Subfamily B metabolism, ATP-Binding Cassette Transporters metabolism, Animals, Base Sequence, Bile Acids and Salts metabolism, Blotting, Western, Carrier Proteins metabolism, Cell Line, DNA Methylation, Glucose metabolism, Glutathione Transferase metabolism, Glycogen metabolism, Hepatocyte Nuclear Factor 3-beta, Hepatocyte Nuclear Factor 6, Homeodomain Proteins metabolism, Immunohistochemistry, Insulin-Like Growth Factor Binding Protein 1 blood, Insulin-Like Growth Factor Binding Protein 1 metabolism, Ligands, Liver embryology, Liver metabolism, Liver pathology, Mice, Mice, Transgenic, Microscopy, Electron, Models, Genetic, Molecular Sequence Data, Organic Anion Transporters, Sodium-Dependent, Phenotype, Prealbumin genetics, Prealbumin metabolism, Promoter Regions, Genetic, Protein Isoforms, Recombinant Proteins metabolism, Symporters, Time Factors, Trans-Activators metabolism, Transcription, Genetic, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Liver cytology, Membrane Transport Proteins, Nuclear Proteins genetics, Nuclear Proteins metabolism, Transcription Factors
- Abstract
The winged helix transcription factor, hepatocyte nuclear factor-3beta (HNF-3beta), mediates the hepatocyte-specific transcription of numerous genes important for liver function. However, the in vivo role of HNF-3beta in regulating these genes remains unknown because homozygous null HNF3beta mouse embryos die in utero prior to liver formation. In order to examine the regulatory function of HNF-3beta, we created transgenic mice in which the -3-kb transthyretin promoter functions to increase hepatocyte expression of the rat HNF-3beta protein. Postnatal transgenic mice exhibit growth retardation, depletion of hepatocyte glycogen storage, and elevated levels of bile acids in serum. The retarded growth phenotype is likely due to a 20-fold increase in hepatic expression of insulin-like growth factor binding protein 1 (IGFBP-1), which results in elevated levels in serum of IGFBP-1 and limits the biological availability of IGFs required for postnatal growth. The defects in glycogen storage and serum bile acids coincide with diminished postnatal expression of hepatocyte genes involved in gluconeogenesis (phosphoenolpyruvate carboxykinase and glycogen synthase) and sinusoidal bile acid uptake (Ntcp), respectively. These changes in gene transcription may result from the disruptive effect of HNF-3beta on the hepatic expression of the endogenous mouse HNF-3alpha,-3beta, -3gamma, and -6 transcription factors. Furthermore, adult transgenic livers lack expression of the canalicular phospholipid transporter, mdr2, which is consistent with ultrastructure evidence of damage to transgenic hepatocytes and bile canaliculi. These transgenic studies represent the first in vivo demonstration that the HNF-3beta transcriptional network regulates expression of hepatocyte-specific genes required for bile acid and glucose homeostasis, as well as postnatal growth.
- Published
- 2000
- Full Text
- View/download PDF
38. Persistent expression of HNF6 in islet endocrine cells causes disrupted islet architecture and loss of beta cell function.
- Author
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Gannon M, Ray MK, Van Zee K, Rausa F, Costa RH, and Wright CV
- Subjects
- Animals, Cell Adhesion, Diabetes Mellitus, Experimental metabolism, Down-Regulation, Endocrine Glands embryology, Eye metabolism, Fluorescent Antibody Technique, Glucose pharmacokinetics, Glucose Tolerance Test, Glucose Transporter Type 2, Glycogen metabolism, Hepatocyte Nuclear Factor 6, Homeodomain Proteins genetics, Immunohistochemistry, Islets of Langerhans cytology, Islets of Langerhans embryology, Liver metabolism, Mice, Mice, Transgenic, Monosaccharide Transport Proteins biosynthesis, Pancreas embryology, Pancreas metabolism, Phenotype, Promoter Regions, Genetic, Radioimmunoassay, Time Factors, Trans-Activators genetics, beta-Galactosidase, Endocrine Glands metabolism, Homeodomain Proteins biosynthesis, Homeodomain Proteins physiology, Islets of Langerhans metabolism, Islets of Langerhans physiology, Trans-Activators biosynthesis, Trans-Activators physiology
- Abstract
We used transgenesis to explore the requirement for downregulation of hepatocyte nuclear factor 6 (HNF6) expression in the assembly, differentiation, and function of pancreatic islets. In vivo, HNF6 expression becomes downregulated in pancreatic endocrine cells at 18. 5 days post coitum (d.p.c.), when definitive islets first begin to organize. We used an islet-specific regulatory element (pdx1(PB)) from pancreatic/duodenal homeobox (pdx1) gene to maintain HNF6 expression in endocrine cells beyond 18.5 d.p.c. Transgenic animals were diabetic. HNF6-overexpressing islets were hyperplastic and remained very close to the pancreatic ducts. Strikingly, alpha, delta, and PP cells were increased in number and abnormally intermingled with islet beta cells. Although several mature beta cell markers were expressed in beta cells of transgenic islets, the glucose transporter GLUT2 was absent or severely reduced. As glucose uptake/metabolism is essential for insulin secretion, decreased GLUT2 may contribute to the etiology of diabetes in pdx1(PB)-HNF6 transgenics. Concordantly, blood insulin was not raised by glucose challenge, suggesting profound beta cell dysfunction. Thus, we have shown that HNF6 downregulation during islet ontogeny is critical to normal pancreas formation and function: continued expression impairs the clustering of endocrine cells and their separation from the ductal epithelium, disrupts the spatial organization of endocrine cell types within the islet, and severely compromises beta cell physiology, leading to overt diabetes.
- Published
- 2000
- Full Text
- View/download PDF
39. Angiotensin converting-like enzymes from urine of untreated renovascular hypertensive and normal patients: purification and characterization.
- Author
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Costa RH, Casarini DE, Plavnik FL, Marson O, and Alves KB
- Subjects
- Adult, Angiotensin-Converting Enzyme Inhibitors pharmacology, Female, Humans, Kinetics, Male, Molecular Weight, Peptidyl-Dipeptidase A isolation & purification, Hypertension, Renovascular enzymology, Peptidyl-Dipeptidase A urine
- Abstract
Angiotensin converting-like enzymes (ACE) were isolated from urine of normal (P0N, P1N and P2N) and untreated renovascular hypertensive (P0, P1 and P2) patients. The urine were submitted to ion exchange chromatography. Enzymes P0 and P0N were eluted with the equilibrium buffer (0.02 M Tris-HCl, pH 7.0), while P1, P1N, P2 and P2N with ionic strength linear gradient of 0.02-0.5 M Tris-HCl, pH 7.0 in 0.7 mS and P2 and P2N in 1.2 mS conductance. The active fractions were submitted to gel filtration in Sephadex G-150, equilibrated and performed with 0.05 M Tris-HCl/0.15 M NaCl buffer, pH 8.0. All enzymes were homogeneous when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (molecular mass: P0, P2 and P2N about 60 kDa; P1, 95 kDa and P21N 170 kDa). The enzymes were recognized by Y1 polyclonal antibody raised against human renal ACE. The K(M) values were in millimolar order for hippuryl-L-His-Leu (HHL) while for benzyloxycarbonyl-Phe-L-His-Leu (ZFHL) they were in 10(-4) M order. The enzymes were able to hydrolyze angiotensin I (AI) (P0 and P0N about 25%, P1 and P1N about 70%, P2 100% and P2N 66%) and bradykinin (BK) (P0N 22%, P1N 81%, P2N 62%, P0 and P1 50% and P2 35%), and their activities were inhibited by captopril.
- Published
- 2000
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40. The nuclear receptor fetoprotein transcription factor is coexpressed with its target gene HNF-3beta in the developing murine liver, intestine and pancreas.
- Author
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Rausa FM, Galarneau L, Bélanger L, and Costa RH
- Subjects
- Animals, Carcinoma, Hepatocellular genetics, DNA-Binding Proteins metabolism, Endoderm, Gene Expression Regulation, Developmental, Hepatocyte Nuclear Factor 3-beta, Intestines growth & development, Liver growth & development, Mice, Nuclear Proteins metabolism, Pancreas growth & development, Receptors, Cytoplasmic and Nuclear genetics, Receptors, Cytoplasmic and Nuclear metabolism, Transcription Factors metabolism, Tumor Cells, Cultured, DNA-Binding Proteins genetics, Intestines embryology, Liver embryology, Nuclear Proteins genetics, Pancreas embryology, Transcription Factors genetics
- Abstract
During organogenesis, the winged helix hepatocyte nuclear factor 3beta (HNF-3beta) protein participates in regulating gene transcription in the developing esophagus, trachea, liver, lung, pancreas, and intestine. Hepatoma cell transfection studies identified a critical HNF-3beta promoter factor, named UF2-H3beta, and here, we demonstrate that UF2-H3beta is identical to the fetoprotein transcription factor (FTF). In situ hybridization studies of mouse embryos demonstrate that FTF expression initiates in the foregut endoderm during liver and pancreatic morphogenesis (day 9) and that earlier expression of FTF is observed in the yolk sac endoderm, branchial arch and neural crest cells (day 8). Abundant FTF hybridization signals are observed throughout morphogenesis of the liver, pancreas, and intestine and its expression continues in the epithelial cells of these adult organs. In day 17 mouse embryos and adult pancreas, however, expression of FTF becomes restricted to the exocrine acinar and ductal epithelial cells.
- Published
- 1999
- Full Text
- View/download PDF
41. Premature expression of the winged helix transcription factor HFH-11B in regenerating mouse liver accelerates hepatocyte entry into S phase.
- Author
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Ye H, Holterman AX, Yoo KW, Franks RR, and Costa RH
- Subjects
- Animals, CCAAT-Enhancer-Binding Proteins, Cell Nucleus metabolism, Cyclins biosynthesis, Cyclins genetics, DNA Replication, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins genetics, Forkhead Box Protein M1, Forkhead Transcription Factors, Humans, Liver cytology, Male, Mice, Mice, Transgenic, Mitosis, Nuclear Proteins biosynthesis, Phosphoproteins biosynthesis, Phosphoproteins genetics, Recombinant Fusion Proteins biosynthesis, Recombinant Fusion Proteins genetics, S Phase, Signal Transduction, Time Factors, Transcription Factors genetics, X-ray Repair Cross Complementing Protein 1, Liver metabolism, Liver Regeneration physiology, Transcription Factors biosynthesis
- Abstract
Two-thirds partial hepatectomy (PH) induces differentiated cells in the liver remnant to proliferate and regenerate to its original size. The proliferation-specific HNF-3/fork head homolog-11B protein (HFH-11B; also known as Trident and Win) is a family member of the winged helix/fork head transcription factors and in regenerating liver its expression is reactivated prior to hepatocyte entry into DNA replication (S phase). To examine whether HFH-11B regulates hepatocyte proliferation during liver regeneration, we used the -3-kb transthyretin (TTR) promoter to create transgenic mice that displayed ectopic hepatocyte expression of HFH-11B. Liver regeneration studies with the TTR-HFH-11B mice demonstrate that its premature expression resulted in an 8-h acceleration in the onset of hepatocyte DNA replication and mitosis. This liver regeneration phenotype is associated with protracted expression of cyclin D1 and C/EBPbeta, which are involved in stimulating DNA replication and premature expression of M phase promoting cyclin B1 and cdc2. Consistent with the early hepatocyte entry into S phase, regenerating transgenic livers exhibited earlier expression of DNA repair genes (XRCC1, mHR21spA, and mHR23B). Furthermore, in nonregenerating transgenic livers, ectopic HFH-11B expression did not elicit abnormal hepatocyte proliferation, a finding consistent with the retention of the HFH-11B transgene protein in the cytoplasm. We found that nuclear translocation of the HFH-11B transgene protein requires mitogenic signalling induced by PH and that its premature availability in regenerating transgenic liver allowed nuclear translocation to occur 8 h earlier than in wild type.
- Published
- 1999
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42. HNF-3/forkhead homologue-4 influences lung morphogenesis and respiratory epithelial cell differentiation in vivo.
- Author
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Tichelaar JW, Lim L, Costa RH, and Whitsett JA
- Subjects
- Animals, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors, Cell Differentiation physiology, Cells, Cultured, Epithelial Cells cytology, Epithelial Cells physiology, Gene Expression Regulation, Developmental physiology, Hepatocyte Nuclear Factor 4, Lung cytology, Mice, Mice, Transgenic, Morphogenesis physiology, Respiratory System cytology, DNA-Binding Proteins, Lung embryology, Phosphoproteins physiology, Transcription Factors physiology
- Abstract
HNF-3/forkhead homologue 4 (HFH-4), a transcription factor of the winged helix/forkhead family, is expressed in various tissues including lung, brain, oviduct, testis, and embryonic kidney. In order to test whether the temporospatial expression of HFH-4 influences lung morphogenesis, HFH-4 was expressed in lungs of transgenic mice under control of the surfactant protein C (SP-C) promoter. The morphology of the lungs from SP-C/HFH-4 embryos (day 18 postconception) was distinctly abnormal, and the severity of the alterations correlated with the level of transgene expression as detected by in situ hybridization. At high levels of expression, HFH-4 altered epithelial cell differentiation and inhibited branching morphogenesis. Atypical cuboidal or columnar cells lined the lung periphery of SP-C/HFH-4 transgenic mice. The atypical epithelial cells seen in the SP-C/HFH-4 mice expressed thyroid transcription factor-1 and hepatocyte nuclear factor 3beta (HNF-3beta). However, surfactant proteins SP-B, SP-C, and Clara cell secretory protein, normally produced by nonciliated epithelial cells in lung parenchyma were lacking. beta-Tubulin IV, a marker of ciliated cells, stained the atypical columnar cells produced by expression of high levels of the SP-C/HFH-4 transgene. Ectopic expression of HFH-4 in developing mouse lung altered epithelial cell differentiation and morphology, restricting the expression of markers typical of nonciliated cells of the distal lung parenchyma., (Copyright 1999 Academic Press.)
- Published
- 1999
- Full Text
- View/download PDF
43. Genesis, a Winged Helix transcriptional repressor, has embryonic expression limited to the neural crest, and stimulates proliferation in vitro in a neural development model.
- Author
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Hromas R, Ye H, Spinella M, Dmitrovsky E, Xu D, and Costa RH
- Subjects
- Animals, Cell Differentiation genetics, Cell Division genetics, Embryonic and Fetal Development genetics, Forkhead Transcription Factors, Helix-Turn-Helix Motifs genetics, Mice, Organ Specificity, Trans-Activators genetics, DNA-Binding Proteins genetics, Gene Expression Regulation, Developmental, Neural Crest embryology, Repressor Proteins genetics
- Abstract
A novel repressor of the Winged Helix (formerly HNF-3/Forkhead) transcriptional regulatory family, termed Genesis (also called HFH2), was previously found to be exclusively expressed in primitive embryonic cell lines. In this study in situ cRNA hybridization experiments revealed that Genesis was expressed during embryogenesis only in developing neural crest cells. Its expression diminished upon their terminal differentiation into sympathetic and parasympathetic neurons. Based on that finding, Genesis was retrovirally transduced into pluripotent N-Tera-2 clone D1 (NT2/D1) teratocarcinoma cells, which are a well-described in vitro model of neural development. Retinoic acid (RA) treatment will drive these cells to differentiation toward the neuronal lineage and cause an increase in expression of the cyclin-dependent kinase inhibitor p21 protein, which leads to an inhibition in cellular proliferation. Although RA-induced expression of neuronal differentiation markers was not influenced by forced overexpression of Genesis in NT2-D1 cells, proliferation of Genesis-transduced cells continued following RA treatment. RA was unable to induce the expression of the cyclin-dependent kinase inhibitor p21 in the Genesis-transduced cells, but Go/G1 tumor suppressor p53 expression was induced normally. Therefore, Genesis may play a role in the regulation of primitive neural crest development by preventing terminal quiescence through inhibition of p21 protein expression. These data also lend evidence for the hypothesis that proliferation and differentiation pathways are not irrevocably linked, but can function independently.
- Published
- 1999
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44. HNF-3/forkhead homologue-4 (HFH-4) is expressed in ciliated epithelial cells in the developing mouse lung.
- Author
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Tichelaar JW, Wert SE, Costa RH, Kimura S, and Whitsett JA
- Subjects
- Animals, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins metabolism, Epithelial Cells metabolism, Hepatocyte Nuclear Factor 4, Immunohistochemistry, Mice, Mice, Knockout, Nuclear Proteins biosynthesis, Nuclear Proteins genetics, Phosphoproteins metabolism, Protein Biosynthesis, Thyroid Nuclear Factor 1, Tissue Distribution, Transcription Factors genetics, Transcription Factors metabolism, Tubulin biosynthesis, Tubulin metabolism, Lung embryology, Lung metabolism, Phosphoproteins biosynthesis, Transcription Factors biosynthesis, Uteroglobin
- Abstract
HNF-3/forkhead homologue-4 (HFH-4), a transcription factor of the winged-helix/forkhead family, was detected by immunohistochemistry in tissue of the developing mouse. HFH-4 protein was present in epithelial cells of the lung, trachea, oviduct, and embryonic esophagus, and in ependymal cells lining the spinal column and ventricles of the brain. In lung, trachea, and nose, HFH-4 was expressed in a distinct subset of epithelial cells that also expressed beta-tubulin IV, a ciliated cell marker. Cellular sites of HFH-4 and beta-tubulin IV expression were distinct from that of Clara cell secretory protein (CCSP), which was detected in nonciliated epithelial cells in the conducting airway of the lung. HFH-4 and beta-tubulin IV, but not CCSP, were detected in the respiratory epithelium of thyroid transcription factor-1 (TTF-1) gene-targeted mice. The presence of HFH-4 and beta-tubulin IV in TTF-1 gene-targeted mice demonstrates that differentiation of ciliated epithelium does not require TTF-1. Co-localization of HFH-4 and beta-tubulin IV staining in various tissues during mouse development supports a role for HFH-4 in the differentiation of ciliated cell lineages.
- Published
- 1999
- Full Text
- View/download PDF
45. The -4 kilobase promoter region of the winged helix transcription factor HNF-3alpha gene elicits transgene expression in mouse embryonic hepatic and intestinal diverticula.
- Author
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Clevidence DE, Zhou H, Lau LF, and Costa RH
- Subjects
- Animals, Embryonic and Fetal Development genetics, Hepatocyte Nuclear Factor 3-alpha, Lung embryology, Mice, Mice, Transgenic, Notochord embryology, Organ Specificity, Recombinant Fusion Proteins, DNA-Binding Proteins genetics, Gene Expression Regulation, Developmental genetics, Intestines embryology, Nuclear Proteins genetics, Promoter Regions, Genetic genetics, Transcription Factors genetics
- Abstract
Murine hepatocyte nuclear factor-3alpha (HNF-3alpha) protein is a member of a large family of developmentally regulated transcription factors that share homology in the winged helix/fork head DNA binding domain and participate in embryonic pattern formation. HNF-3alpha also mediates cell-specific transcription of genes important for the function of hepatocytes, intestinal, pancreatic and bronchiolar epithelium. We have previously determined that -520 nucleotides upstream of the rat HNF-3alpha gene were sufficient to elicit hepatoma-specific expression in transfection assays and reported on a novel HNF-3alpha expression pattern in the renal pelvis urothelium of the embryonic and adult kidney. We also showed that retinoic acid mediated activation of the HNF-3alpha gene required -4 kb of the HNF-3alpha promoter region in F9 teratocarcinoma transfections. In order to determine regulatory sequences mediating the HNF-3alpha cellular expression pattern in developing mouse embryos, we created transgenic mice bearing the -4 kb HNF-3alpha promoter region driving expression of the beta-galactosidase transgene. Embryonic analysis of two transgenic mouse lines demonstrated that the -4 kb HNF-3alpha promoter sequences were sufficient to elicit transgene expression in the developing liver, intestine, esophagus, nasal epithelial cells and floorplate of the neurotube, but not in the mesodermal notochord or in the lung bud. One of the transgenic lines also exhibited proper expression in the mesonephric ducts and metanephric diverticulum, suggesting that the -4 kb HNF-3alpha promoter region contained a subset of the regulatory sequences necessary for HNF-3alpha expression in the developing kidney.
- Published
- 1998
46. In situ hybridization with 33P-labeled RNA probes for determination of cellular expression patterns of liver transcription factors in mouse embryos.
- Author
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Rausa FM, Ye H, Lim L, Duncan SA, and Costa RH
- Subjects
- Animals, Cell Differentiation physiology, DNA-Binding Proteins genetics, Gestational Age, Hepatocyte Nuclear Factor 3-beta, Hepatocyte Nuclear Factor 6, Homeodomain Proteins genetics, Intestines embryology, Liver embryology, Mice, Nuclear Proteins genetics, Oligonucleotides, Antisense genetics, Pancreas embryology, Phosphorus Isotopes, Prealbumin genetics, Promoter Regions, Genetic, RNA, Messenger analysis, Trans-Activators genetics, Gene Expression Regulation, Developmental genetics, In Situ Hybridization, Fluorescence methods, RNA Probes genetics, Transcription Factors analysis
- Abstract
Murine hepatocyte nuclear factor-3beta (HNF-3beta) protein is a member of a large family of developmentally regulated transcription factors that share homology in the winged helix/fork head DNA binding domain and that participate in embryonic pattern formation. HNF-3beta also mediates cell-specific transcription of genes important for the function of hepatocytes, intestinal and bronchiolar epithelium, and pancreatic acinar cells. We have previously identified a hepatocyte and pancreatic cut-homeodomain transcription factor, HNF-6, which is required for HNF-3beta promoter activity. In this study, we used in situ hybridization studies of stage-specific embryos to demonstrate that HNF-6 and its target gene, HNF-3beta, are coexpressed in the foregut endoderm and in the pancreatic and hepatic diverticulum. More detailed analysis of HNF-6 and HNF-3beta's developmental expression patterns provides evidence of colocalization in hepatocytes, intestinal epithelium, and pancreatic ductal epithelium and exocrine acinar cells. In support of the role of HNF-6 in regulating HNF-3beta expression in developing hepatocytes, their liver expression levels are both transiently reduced between 14 and 15 days of gestation. At day 18 of gestation and in adult pancreas, HNF-6 and HNF-3beta transcripts remain colocalized in the exocrine acinar cells, but their expression patterns diverge in endocrine cells. HNF-3beta expression is restricted to the endocrine cells of the islets of Langerhans, whereas the ductal epithelium expresses HNF-6. We discuss these expression patterns with respect to specification of hepatocytes and differentiation of the endocrine and exocrine pancreas., (Copyright 1998 Academic Press.)
- Published
- 1998
- Full Text
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47. Differential regulation of interleukin-8 and intercellular adhesion molecule-1 by H2O2 and tumor necrosis factor-alpha in endothelial and epithelial cells.
- Author
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Lakshminarayanan V, Beno DW, Costa RH, and Roebuck KA
- Subjects
- Animals, Chemokine CCL5 metabolism, Culture Media, Conditioned, Humans, Intercellular Adhesion Molecule-1 genetics, Interleukin-8 genetics, RNA, Messenger metabolism, Tumor Cells, Cultured, Endothelium, Vascular metabolism, Epithelial Cells metabolism, Hydrogen Peroxide metabolism, Intercellular Adhesion Molecule-1 metabolism, Interleukin-8 metabolism, Tumor Necrosis Factor-alpha metabolism
- Abstract
The reactive oxygen intermediate H2O2 can function as a signaling molecule to activate gene expression. In this study, we demonstrate that oxidant stress induced by tumor necrosis factor alpha (TNFalpha) or H2O2 differentially regulates intercellular adhesion molecule-1 (ICAM-1) and interleukin-8 (IL-8) gene expression in endothelial and epithelial cells. Northern blot analysis revealed that TNFalpha induced both ICAM-1 and IL-8 expression in either the A549 lung epithelial cell line or the human microvessel endothelial cell line (HMEC-1). In contrast, H2O2 selectively induced only ICAM-1 in HMEC-1 and only IL-8 in A549. This cell type-specific pattern of IL-8 expression was also observed in several other endothelial and epithelial cells. TNFalpha induced greater IL-8 gene expression as compared with H2O2, but the kinetics of induction were similar. The induction of epithelial IL-8 message was accompanied by a corresponding increase in functional IL-8 protein secretion as determined by a neutrophil motility assay. The increased neutrophil motility stimulated by conditioned media from H2O2- or TNFalpha-exposed A549 cells was completely inhibited by an anti-IL-8 antibody. TNFalpha and H2O2 also induced a differential pattern of CC chemokine expression in A549. While TNFalpha induced both RANTES and MCP-1, H2O2 induced only MCP-1. These data suggest that epithelial cells under oxidant stress contribute to the inflammatory cytokine network by selective production of IL-8, MCP-1, and RANTES, which may critically influence the site-specific recruitment of leukocyte subsets.
- Published
- 1997
- Full Text
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48. The cut-homeodomain transcriptional activator HNF-6 is coexpressed with its target gene HNF-3 beta in the developing murine liver and pancreas.
- Author
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Rausa F, Samadani U, Ye H, Lim L, Fletcher CF, Jenkins NA, Copeland NG, and Costa RH
- Subjects
- Amino Acid Sequence, Animals, Carcinoma, Hepatocellular pathology, Cell Differentiation, Crosses, Genetic, DNA-Binding Proteins biosynthesis, Female, Fetal Proteins biosynthesis, Fetal Proteins genetics, Hepatocyte Nuclear Factor 3-beta, Hepatocyte Nuclear Factor 6, Homeodomain Proteins biosynthesis, Homeodomain Proteins genetics, Humans, In Situ Hybridization, Intestinal Mucosa embryology, Intestinal Mucosa metabolism, Islets of Langerhans embryology, Islets of Langerhans metabolism, Liver metabolism, Liver Neoplasms pathology, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Multigene Family, Muridae genetics, Nuclear Proteins biosynthesis, Organ Specificity, Pancreas metabolism, Promoter Regions, Genetic, Sequence Alignment, Sequence Homology, Amino Acid, Trans-Activators biosynthesis, Trans-Activators genetics, Transcription, Genetic, Tumor Cells, Cultured, DNA-Binding Proteins genetics, Fetal Proteins physiology, Gene Expression Regulation, Developmental, Genes, Homeobox, Homeodomain Proteins physiology, Liver embryology, Nuclear Proteins genetics, Pancreas enzymology, Trans-Activators physiology, Transcription Factors
- Abstract
Murine hepatocyte nuclear factor-3 beta (HNF-3 beta) protein is a member of a large family of developmentally regulated transcription factors that share homology in the winged helix/fork head DNA binding domain and that participate in embryonic pattern formation. HNF-3 beta also mediates cell-specific transcription of genes important for the function of hepatocytes, intestinal and bronchiolar epithelial, and pancreatic acinar cells. We have previously identified a liver-enriched transcription factor, HNF-6, which is required for HNF-3 beta promoter activity and also recognizes the regulatory region of numerous hepatocyte-specific genes. In this study we used the yeast one-hybrid system to isolate the HNF-6 cDNA, which encodes a cut-homeodomain-containing transcription factor that binds with the same specificity as the liver HNF-6 protein. Cotransfection assays demonstrate that HNF-6 activates expression of a reporter gene driven by the HNF-6 binding site from either the HNF-3 beta or transthyretin (TTR) promoter regions. We used interspecific backcross analysis to determine that murine Hnf6 gene is located in the middle of mouse chromosome 9. In situ hybridization studies of staged specific embryos demonstrate that HNF-6 and its potential target gene, HNF-3 beta, are coexpressed in the pancreatic and hepatic diverticulum. More detailed analysis of HNF-6 and HNF-3 beta's developmental expression patterns provides evidence of colocalization in hepatocytes, intestinal epithelial, and in the pancreatic ductal epithelial and exocrine acinar cells. The expression patterns of these two transcription factors do not overlap in other endoderm-derived tissues or the neurotube. We also found that HNF-6 is also abundantly expressed in the dorsal root ganglia, the marginal layer, and the midbrain. At day 18 of gestation and in the adult pancreas, HNF-6 and HNF-3 beta transcripts colocalize in the exocrine acinar cells, but their expression patterns diverge in other pancreatic epithelium. HNF-6, but not HNF-3 beta, expression continues in the pancreatic ductal epithelium, whereas only HNF-3 beta becomes restricted to the endocrine cells of the islets of Langerhans. We discuss these expression patterns with respect to specification of hepatocytes and differentiation of the endocrine and exocrine pancreas.
- Published
- 1997
- Full Text
- View/download PDF
49. The winged helix transcriptional activator HFH-8 is expressed in the mesoderm of the primitive streak stage of mouse embryos and its cellular derivatives.
- Author
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Peterson RS, Lim L, Ye H, Zhou H, Overdier DG, and Costa RH
- Subjects
- Animals, Binding Sites, Capillaries metabolism, Cell Line, Digestive System embryology, Endothelium cytology, Endothelium metabolism, Fibroblasts metabolism, Forkhead Transcription Factors, Helix-Turn-Helix Motifs, Humans, Intestinal Mucosa metabolism, Intestines embryology, Intestines growth & development, Lung blood supply, Lung embryology, Lung growth & development, Mesoderm metabolism, Mice, Muscle, Smooth metabolism, DNA-Binding Proteins, Embryo, Mammalian physiology, Gene Expression Regulation, Developmental, Mesoderm physiology, Trans-Activators genetics, Trans-Activators metabolism
- Abstract
The hepatocyte nuclear factor 3/fork head homolog (HFH) proteins are an extensive family of transcription factors which share homology in the winged helix DNA binding domain. Members of the winged helix family have been implicated in cell fate determination during pattern formation, in organogenesis and in cell type-specific gene expression. In this study, we used in situ hybridization to identify the cellular expression pattern of the winged helix transcription factor, HFH-8, during mouse embryonic development. We showed that HFH-8 expression initiates during the primitive streak stage of mouse embryogenesis in the extraembryonic mesoderm and in the lateral mesoderm which gives rise to the somatopleuric and splanchnopleuric mesoderm. During organogenesis, HFH-8 expression is found in the splanchnic mesoderm in close apposition of the gut endoderm, suggesting a role in mesenchymal-epithelial induction of lung and gut morphogenesis. HFH-8 expression continues in lateral mesoderm-derived tissue throughout mouse development. HFH-8 expression is observed in the mesenchymal cells of the oral cavity, esophagus, trachea, lung, intestine, dorsal aorta and intersomitic arteries, but not in the vasculature of the head, liver, kidney or heart. Consistent with these embryonic expression studies, adult HFH-8 expression is restricted to the endothelium and connective fibroblasts of the alveolar sac and in the lamina propria and smooth muscle of the intestine. We also show that several adult endothelial cell lines maintain abundant HFH-8 expression. Furthermore, we used our determined HFH-8 consensus sequence to identify putative target genes expressed in pulmonary and intestinal mesenchymal cells. Cotransfection assays with one of these target promoters, P-selectin, demonstrated that HFH-8 expression was required for IL-6 stimulation of P-selectin promoter activity and suggest that HFH-8 is involved in mediating its cell-specific transcriptional activation in response to cytokines.
- Published
- 1997
- Full Text
- View/download PDF
50. Hepatocyte nuclear factor-3beta limits cellular diversity in the developing respiratory epithelium and alters lung morphogenesis in vivo.
- Author
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Zhou L, Dey CR, Wert SE, Yan C, Costa RH, and Whitsett JA
- Subjects
- Animals, Cadherins genetics, Cell Differentiation, DNA-Binding Proteins genetics, Endothelial Growth Factors genetics, Hepatocyte Nuclear Factor 3-beta, Humans, Lung cytology, Lymphokines genetics, Mesoderm cytology, Mice, Mice, Transgenic, Morphogenesis, Nuclear Proteins genetics, RNA, Messenger metabolism, Rats, Transcription Factors genetics, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factors, DNA-Binding Proteins physiology, Epithelial Cells cytology, Lung embryology, Nuclear Proteins physiology, Transcription Factors physiology
- Abstract
Hepatocyte nuclear factor-3beta (HNF-3beta), a nuclear protein of the winged helix family of transcription factors, is known to play a critical role in the formation of the embryonic node, notochord, and foregut endoderm. HNF-3beta influences the expression of a number of target genes in the respiratory epithelium, activating transcription of thyroid transcription factor-1, surfactant protein-B and clara cell secretory protein. In order to discern the role of HNF-3beta in differentiation and gene expression in the lung, HNF-3beta was expressed in developing respiratory epithelial cells of transgenic mice, under the control of the human surfactant protein C gene promoter. Pulmonary abnormalities were observed in the lungs of fetal mice bearing the HNF-3beta transgene. Differentiation of distal respiratory epithelial cells was arrested in the early pseudoglandular stage. Branching morphogenesis and vasculogenesis were markedly disrupted in association with decreased E-cadherin and vascular endothelial growth factor expression. HNF-3beta limits cellular diversity of developing respiratory epithelium and alters lung morphogenesis in vivo, suggesting that precise temporal-spatial regulation of HNF-3beta expression is critical for respiratory epithelial cell differentiation and lung morphogenesis.
- Published
- 1997
- Full Text
- View/download PDF
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