Bioconversion of Cellulose to Hydrogen by dark fermentation

Hydrogen is known as a clean energy resource. The biological production of hydrogen has been attracting attention as an environmentally friendly processs that does not consume fossil fuels. Cellulosic plant and waste materials are potential resources for fermentative hydrogen production. Cellulose is a linear biopolymer of glucose molecules, connected by β-1,4-glycosidic bonds. Enzymatic hydrolysis of cellulose requires the presence of cellulase. The present study aimed to investigate the efficiency of acid pretreatment on ruminal fluid in order to enrich H2 producing bacteria consortia to enhance biohydrogen rate and substrate removal efficiency. In this study, fermentative hydrogen producers were enriched on cellulose (2g/L) in a modificated Del Nery medium (DNM) at 37ºC and initial pH 7.0 using rumen fluid (10% v/v) as inoculum. To increase the hydrogen production it was added cellulose (10mL) to the medium. The gas products (mainly H2 and CO2) was analyzed by gas chromatography (Shimadzu GC 2010) using a thermal conductivity detector. The volatile fatty acids and ethanol were also detected by GC using a flame ionization detector. Cellulose degradation was quantified by using the phenolsulfuric acid method. Analysis showed that the biogas produced from the anaerobic fermentation contained only hydrogen and carbon dioxide, without detectable methane after acid pretreatment test. On DNM the hydrogen production started with 4 h (5,3 x 105 mmol H2/L) of incubation, and the maximum H2 concentration was observed with 34 h (7,1 x 106 mmol H2/L) of incubation. During the process, it was observed a predominance of acetic acid and butyric acid as well as a low production of acetone, ethanol and nbutanol in all experimental phases. Butyrate accounted for more than 77% of total. As a result of the accumulation of volatile fatty acids (VFAs), the pH value in anaerobic digestion system was reduced to 4,0. On microscopy analyses there were observed rods with endospores. The batch anaerobic fermentation assays performed on anaerobic mixed inoculum from rumen fluid demonstrated the feasibility of H2 generation utilizing cellulose as substrate. Based on the results, it can be concluded that the acid treatment was efficient to inhibit the methanogenic archaea cells present in rumen fluid. The rumen fluid cells present a potential route in converting renewable biomass such as cellulose into hydrogen energy.

Biological Hydrogen production from environmental sample in tropical countries

The hydrogen gas is regarded as clean and renewable energy source, since it generates only water during combustion when used as fuel. It shows 2.75 times more energy content than any hydrocarbon and it can be converted into electrical, mechanical energy or heat. Inoculum sources have been successfully tested for hydrogen biological production in temperate climate countries as sludge treatment plants sewage, sludge treatment plant wastewater, landfill sample, among others. However, hydrogen biologic production with inoculum from environmental samples such as sediment reservoirs, especially in tropical countries like Brazil, is rarely investigated. Reservoirs and fresh water lake sediment may contain conditions for the survival of a wide variety of microorganisms which use different carbon sources mainly glucose and xylose, in the fermentation. Glucose is an easily biodegradable, present in most of the industrial effluents and can be obtained abundantly from agricultural wastes. A wide variety of wastewater resulting from agriculture, industry and pulp and paper processed from wood may contain xylose in its constitution. Such effluent contains glucose and xylose concentrations of about 2 g/L. In this sense, this work verified hydrogen biological production in anaerobic batch reactor (1L), at 37 ° C, initial pH 5.5, headspace with N2 (100%), Del Nery medium, vitamins and peptone (1 g/L), fed separately with glucose (2g/L) and xylose (2 g/L). The inoculum was taken from environmental sample (sediment reservoir Itupararanga - Ibiúna - SP-Brazil). It was previously purified in serial dilutions at H2 generation (10-5, 10-7, 10-10), and heat treated (90º C - 10 min) later to inhibited the H2 consumers. The maximum H2 generations obtained in both tests were observed at 552 h, as described below. At the reactors fed with glucose and xylose were observed, respectively, 9.1 and 8.6 mmol H2/L, biomass growth (0.2 and 0.2 nm); consumption of sugar concentrations 53.6% (1.1 glucose g/L) and 90.5% (1.8 xylose g/L); acetic acid generation (124.7 mg/L and 82.7 mg/L), butyric acid (134.0 mg/L and 230.4 mg/L) and there wasn’t methane generation in the reactors. Microscopic analysis of biomass in anaerobic reactors showed the predominance of Gram positive rods and rods with endospores, whose morphology is characteristic of H2-generating bacteria, in both tests. These species were selected from the natural environment. In DGGE analysis performed difference were observed between populations from inoculum and in tests. This analysis confirmed that some species of bacteria were selected which remained under the conditions imposed on the experiment. The efficiency of the pre-treatment of inoculum and the imposition of pH 5.5 inhibited methane-producing microorganisms and the consumers of H2. Therefore, the experimental conditions imposed allowed the attainment of bacterial consortium of producer H2 taken from an environmental sample with concentration of xylose and glucose similar to the ones of the industrial effluents.

Low toxic effects of a whitening strip to cultured pup cells

Fundacao de Amparo a Pesquisa do Estado de sao Paulo (FAPESP)

Immediate and late analysis of dental pulp stem cells viability after indirect exposition to alternative in-office bleaching strategies

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Responses of dental pulp cells to a less invasive bleaching technique applied to adhesive restored teeth

To assess the cytotoxicity of 35% hydrogen peroxide (HP) bleaching gel applied for 15 min to sound or restored teeth with two-step self-etching adhesive systems and composite resin. Materials and Methods: Sound and restored enamel/dentin disks were stored in water for 24 h or 6 months + thermocycling. The disks were adapted to artificial pulp chambers and placed in compartments containing culture medium. Immediately after bleaching, the culture medium in contact with dentin was applied for 1 h to previously cultured odontoblast-like MDPC-23 cells. Thereafter, cell viability (MTT assay) and morphology (SEM) were assessed. Data were analyzed by two-way ANOVA and Tukey's test (a = 5%). Results: In comparison to the negative control group (no treatment), no significant cell viability reduction occurred in those groups in which sound teeth were bleached. However, a significant decrease in cell viability was observed in the adhesive-restored bleached groups compared to negative control. No significant difference among bleached groups was observed with respect to the presence of restoration and storage time. Conclusion: The application of 35% HP bleaching gel to sound teeth for 15 min does not cause toxic effects in pulp cells. When this bleaching protocol was performed in adhesive-restored teeth, a significant toxic effect occurred.

Transdentinal cytotoxicity of carbodiimide (EDC) and glutaraldehyde on odontoblast-like cells

Objective: To evaluate the transdentinal cytotoxicity of three different concentrations of carbodiimide (EDC) or 5% glutaraldehyde (GA) on MDPC-23 cells. Methods: Seventy 0.4-mm-thick dentin disks obtained from human molars were adapted to artificial pulp chambers. MDPC-23 cells were seeded on the pulpal surface of the disks. After 48 hours, the occlusal dentin was acid-etched and treated for 60 seconds with one of the following solutions (n=10): no treatment (negative control); 0.1 M, 0.3 M, or 0.5 M EDC; 5% GA; Sorensen buffer; or 29% hydrogen peroxide (positive control). Cell viability and morphology were assessed by methyltetrazolium assay and scanning electron microscopy (SEM), respectively. The eluates were collected after the treatments and applied on MDPC-23 seeded in a 24-well plate to analyze cell death, total protein (TP), and collagen production. The last two tests were performed 24 hours and seven days after the challenge. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). Results: EDC at all test concentrations did not reduce cell viability, while 5% GA did increase cell metabolism. Cell death by necrosis was not elicited by EDC or 5% GA. At the 24-hour period, 0.3 M and 0.5 M EDC reduced TP production by 18% and 36.8%, respectively. At seven days, increased TP production was observed in all groups. Collagen production at the 24-hour period was reduced when 0.5 M EDC was used. After seven days, no difference was observed among the groups. SEM showed no alteration in cell morphology or number, except in the hydrogen peroxide group. Conclusions: Treatment of acid-etched dentin with EDC or GA did not cause transdentinal cytotoxic effects on odontoblast-like cells.

Color alteration, hydrogen peroxide diffusion, and cytotoxicity caused by in-office bleaching protocols

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hydrogen peroxide attenuates the dipsogenic, renal and pressor responses induced by cholinergic activation of the medial septal area

Cholinergic activation of the medial septal area (MSA) with carbachol produces thirst, natriuresis, antidiuresis and pressor response. In the brain, hydrogen peroxide (H2O2) modulates autonomic and behavioral responses. In the present study, we investigated the effects of the combination of carbachol and H2O2 injected into the MSA on water intake, renal excretion, cardiovascular responses and the activity of vasopressinergic and oxytocinergic neurons in the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. Furthermore, the possible modulation of carbachol responses by H2O2 acting through K+ATP channels was also investigated. Male Holtzman rats (280–320 g) with stainless steel cannulas implanted in the MSA were used. The pre-treatment with H2O2 in the MSA reduced carbachol-induced thirst (7.9 ± 1.0, vs. carbachol: 13.2 ± 2.0 ml/60 min), antidiuresis (9.6 ± 0.5, vs. carbachol: 7.0 ± 0.8 ml/120 min,), natriuresis (385 ± 36, vs. carbachol: 528 ± 46 μEq/120 min) and pressor response (33 ± 5, vs. carbachol: 47 ± 3 mmHg). Combining H2O2 and carbachol into the MSA also reduced the number of vasopressinergic neurons expressing c-Fos in the PVN (46.4 ± 11.2, vs. carbachol: 98.5 ± 5.9 c-Fos/AVP cells) and oxytocinergic neurons expressing c-Fos in the PVN (38.5 ± 16.1, vs. carbachol: 75.1 ± 8.5 c-Fos/OT cells) and in the SON (57.8 ± 10.2, vs. carbachol: 102.7 ± 7.4 c-Fos/OT cells). Glibenclamide (K+ATP channel blocker) into the MSA partially reversed H2O2 inhibitory responses. These results suggest that H2O2 acting through K+ATP channels in the MSA attenuates responses induced by cholinergic activation in the same area.

Responses of Dental Pulp Cells to a Less Invasive Bleaching Technique Applied to Adhesively Restored Teeth

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Responses of human dental pulp cells after application of a low-concentration bleaching gel to enamel

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)