Influência do plasma rico em plaquetas no reparo ósseo de defeitos de tamanho crítico em calvárias de ratos: estudo histológico e histométrico

Pós-graduação em Odontologia - FOA

Influência do plasma rico em plaquetas associado ou não à regeneração tecidual guiada na cicatrização de defeitos de fenestração periodontal em cães: estudo histológico e histométrico

Pós-graduação em Odontologia - FOA

Influência da associação do plasma rico em plaquetas ao enxerto de osso autógeno no reparo ósseo de defeitos de tamanho crítico em calvárias de coelhos: estudo histológico e histométrico

Pós-graduação em Odontologia - FOA

Gene therapy strategies for the treatment of spinal cord injury

Spinal cord injury is a complex pathology often resulting in functional impairment and paralysis. Gene therapy has emerged as a possible solution to the problems of limited neural tissue regeneration through the administration of factors promoting axonal growth, while also offering long-term local delivery of therapeutic molecules at the injury site. Of note, gene therapy is our response to the requirements of neural and glial cells following spinal cord injury, providing, in a time-dependent manner, growth substances for axonal regeneration and eliminating axonal growth inhibitors. Herein, we explore different gene therapy strategies, including targeting gene expression to modulate the presence of neurotrophic growth or survival factors and increase neural tissue plasticity. Special attention is given to describing advances in viral and non-viral gene delivery systems, as well as the available routes of gene delivery. Finally, we discuss the future of combinatorial gene therapies and give consideration to the implementation of gene therapy in humans. © 2014 Future Science Ltd.

Lactofem e etefom como reguladores de crescimento de cultivares de soja.

2016

Endophytic and rhizospheric enterobacteria isolated from sugar cane have different potentials for producing plant growth-promoting substances

Background and aims Endophytic and rhizospheric environments differ in many respects, leading to the presence of different bacterial communities at each site. However, microorganisms such as enterobacteria can be found both within plants and in the surrounding soil. Bacteria must present differences in the traits that affect such environments in order to successfully colonise them. The present study compared the plant growth-promoting potential of diazotrophic enterobacteria isolated from the rhizosphere and from within surface-disinfected plants. Methods A total of 46 diazotrophic enterobacterial strains (21 rhizospheric and 25 putatively endophytic) belonging to the Klebsiella and Enterobacter genera, which are prevalent in sugar cane plantations, were isolated from the rhizosphere and from surface-disinfected plants. Their ability to synthesise amino acids using combined nitrogen obtained from nitrogen fixation, and their ability to synthesise indole-3-acetic acid (IAA) were determined by high performance liquid chromatography. Endogenous ethylene production by the bacteria was measured using gas chromatography, and biocontrol of phytopathogenic fungi was determined qualitatively using a dual culture technique. Results The putative endophytes released significantly higher amounts of amino acids than the rhizospheric bacteria, whilst the latter produced higher quantities of ethylene and were more actively antagonistic to fungi. Both types of bacteria released similar amounts of IAA. Conclusion Endophytic and rhizospheric bacteria differ in their capacity to release plant growth-promoting substances, which may be a reflection of their adaptations and an indication of their potential impact on their natural environment.

Effects of potentised substances on growth kinetics of Saccharomyces cerevisiae and Schizosaccharomyces pombe

BACKGROUND: Homeopathic potencies are used as specific remedies in complementary medicine. Since the mode of action is unknown, the presumed specificity is discussed controversially. OBJECTIVE: This study investigated the effects of potentised substances on two yeast species, Saccharomyces cerevisiae and Schizosaccharomyces pombe, in a stable and reliable test system with systematic negative controls. MATERIALS AND METHODS: Yeast cells were cultivated in either potentised substances or water controls in microplates and their growth kinetics were measured photometrically. Water control runs were performed repeatedly to investigate the stability of the experimental set-up (systematic negative controls). RESULTS: 4 out of 14 screened substances seem to have affected the growth curve parameters slope or yield. Out of these substances, azoxystrobin and phosphorus were chosen for 8 further replication experiments, which partly confirmed the results of the screening. On the average of all experiments, azoxystrobin affected the slope of the growth curve of Saccharomyces cerevisiae (p < 0.05), and phosphorus affected the slope of the growth curve of Schizosaccharomyces pombe (p < 0.05). No effects were seen in the water control runs. In addition, significant interactions between treatment with potentised substances and experiment number were observed in all experiments with potentised substances (p < 0.01), but not in the water control runs. CONCLUSIONS: Both yeast species reacted to certain potentised substances by changing their growth kinetics. However, the interactions found point to additional factors of still unknown nature, that modulate the effects of potentised substances. This stable test system with yeasts may be suitable for further studies regarding the efficacy of homeopathic potencies.

The mechanism of maturation of insulin-like growth factor-1

This study, to elucidate the role of des(1-3)IGF-I in the maturation of IGF-I,used two strategies. The first was to detect the presence of enzymes in tissues, which would act on IGF-I to produce des(1-3)IGF-I, and the second was to detect the potential products of such enzymic activity, namely Gly-Pro-Glu(GPE), Gly-Pro(GP) and des(l- 3)IGF-I. No neutral tripeptidyl peptidase (TPP II), which would release the tripeptide GPE from IGF-I, was detected in brain, urine nor in red or white blood cells. The TPPlike activity which was detected, was attributed to a combined action of a dipeptidyl peptidase (DPP N) and an aminopeptidase (AP A). A true TPP II was, however, detected in platelets. Two purified TPP II enzymes were investigated but they did not release GPE from IGF-I under a variety of conditions. Consequently, TPP II seemed unlikely to participate in the formation of des(1-3)IGF-I. In contrast, an acidic tripeptidyl peptidase activity (TPP I) was detected in brain and colostrum, the former with a pH optimum of 4.5 and the latter 3.8. It seems likely that such an enzyme would participate in the formation of des( 1-3 )IGF-I in these tissues in vitro, ie. that des(1-3)IGF-I may have been produced as an artifact in the isolation of IGF-I from brain and colostrum in acidic conditions. This contrasts with suggestions of an in vivo role for des(1-3)IGF-I, as reported by others. The activity of a dipeptidyl peptidase N (DPP N) from urine, which should release the dipeptide GP from IGF-I, was assessed under a variety of conditions and with a variety of additives and potential enzyme stimulants, but there was no release of GP. The DPP N also exhibited a transferase activity with synthetic substrates in the presence of dipeptides, at lower concentrations than previously reported for other acceptors or other proteolytic enzymes. In addition, a low concentration of a product,possibly the tetrapeptide Gly-Pro-Gly-Leu, was detected with the action of the enzyme on IGF-I in the presence of the dipeptide Gly-Leu. As part of attempts to detect tissue production of des(1-3)IGF-I, a monoclonal antibody (MAb ), directed towards the GPE- end ofiGF-I was produced by immunisation with a 10-mer covalently attached to a carrier protein. By the use of indirect ELISA and inhibitor studies, the MAb was shown to selectively recognise peptides with anNterminal GPE- sequence, and applied to the indirect detection of des(1-3)IGF-I. The concentration of GPE in brain, measured by mass spectrometry ( MS), was low, and the concentration of total IGF-I (measured by ELISA with a commercial polyclonal antibody [P Ab]) was 40 times higher at 50 nmol/kg. This also, was not consistent with the action of a tripeptidyl peptidase in brain that converted all IGF-I to des(1-3)IGF-I plus GPE. Contrasting ELISA results, using the MAb prepared in this study, suggest an even higher concentration of intact IGF-I of 150 nmollkg. This would argue against the presence of any des( 1-3 )IGF-I in brain, but in turn, this indicates either the presence of other substances containing a GPE amino-terminus or other cross reacting epitope. Although the results of the specificity studies reported in Chapter 5 would make this latter possibility seem unlikely, it cannot be completely excluded. No GP was detected in brain by MS. No GPE was detected in colostrum by capillary electrophoresis (CE) but the interference from extraneous substances reduced the detectability of GPE by CE and this approach would require further, prior, purification and concentration steps. A molecule, with a migration time equal to that of the peptide GP, was detected in colostrum by CE, but the concentration (~ 10 11mo/L) was much higher than the IGF-I concentration measured by radio-immunoassay using a PAb (80 nmol/L) or using a Mab (300-400 nmolL). A DPP IV enzyme was detected in colostrum and this could account for the GP, derived from substrates other than IGF-1. Based on the differential results of the two antibody assays, there was no indication of the presence of des(1-3)IGF-I in brain or colostrum. In the absence of any enzyme activity directed towards the amino terminus of IGF-I and the absence any potential products, IGF-I, therefore, does not appear to "mature" via des(1-3)IGF-I in the brain, nor in the neutral colostrum. In spite of these results which indicate the absence of an enzymic attack on IGF-I and the absence of the expected products in tissues, the possibility that the conversion of IGF-I may occur in neutral conditions in limited amounts, cannot be ruled out. It remains possible that in the extracellular environment of the membrane, a complex interaction of IGF-I, binding protein, aminopeptidase(s) and receptor, produces des(1- 3)IGF-I as a transient product which is bound to the receptor and internalised.

Autocalibration of dynamic bacterial growth model for water distribution system using GA

The presence of residual chlorine and organic matter govern the bacterial regrowth within a water distribution system. The bacterial growth model is essential to predict the spatial and temporal variation of all these substances throughout the system. The parameters governing the bacterial growth and biodegradable dissolved organic carbon (BDOC) utilization are difficult to determine by experimentation. In the present study, the estimation of these parameters is addressed by using simulation-optimization procedure. The optimal solution by genetic algorithm (GA) has indicated that the proper combination of parameter values are significant rather than correct individual values. The applicability of the model is illustrated using synthetic data generated by introducing noise in to the error-free measurements. The GA was found to be a potential tool in estimating the parameters controlling the bacterial growth and BDOC utilization. Further, the GA was also used for evaluating the sensitivity issues relating parameter values and objective function. It was observed that mu and k(cl) are more significant and dominating compared to the other parameters. But the magnitude of the parameters is also an important issue in deciding the dominance of a particular parameter. GA is found to be a useful tool in autocalibration of bacterial growth model and a sensitivity study of parameters.

Study of effect Rosmarinus officinalis oil and nisin on the growth of Streptococcus iniae in lab conditions and fillets of rainbow trout (Oncorhynchus mykiss)

Fish are an important part of a healthy diet since they contain high quality protein, but typically present a low fat percent when compared to other meats. Fish is an extremely perishable food commodity. On the other hand, food borne diseases are still a major problem in the world, even in well-developed countries. The increasing incidence of food borne diseases coupled with the resultant social and economic implications means there is a constant striving to produce safer food and to develop new antimicrobial agents concerns over the safety of some chemical preservatives and negative consumer reactions to preservatives they perceive as chemical and artificial, have prompted on increased interest in more ‘‘naturalgreen’’ alternatives for the maintenance or extension of product shelf-life. Particular interest has focused on the potential applications of plant essential oils. However, to establish the usefulness of natural antimicrobial preservatives, they must be evaluated alone and in combination with other preservation factors to determine whether there are synergistic effects and multiple hurdles can be devised. In this study, were evaluated the effects of different concentrations of Rosmarinus officinalis and nisin and storage time (15 days) on growth of Streptococcus iniae GQ850377 in a lab conditions and a food model system (fillets of rainbow trout) in 4 and 8 °C. In addition, we also studied multi factorial effects of four different concentration of rosemary, three different concentrations of nisin, two different levels of pH in 3 temperature 4,15 and 37 °C on log% of S.iniae during 43 days in BHI broth. The results on growth of S. iniae were evaluated using SPSS 20.0 statistical software and analyzed the logarithm of total count of the bacterial by Tukey Test. Results were considered statistically significant when P<0.05. MIC and MBC values of rosemary and nisin were 0.03, 0.075 % and 5, 40 μg/mL, respectively. The growth of S. iniae was effected significantly (P<0.05) by rosemary and nisin and also combination of rosemary and nisin in 4 and 8 °C. Samples treated with 0.135 and 0.405 % of rosemary showed a significant decrease on the growth of the bacteria compared with control sample(P<0.05). The most ١٤٦ inhibitory effects were seen in samples treated with 0.135 and 0.405% of rosemary until 9 days after storage. Also, the synergism effects of rosemary and nisin on the growth rate of bacteria was significant (P<0.05) compared with untreated samples and samples treated with the rosemary or nisin, only. Synergistic effects was observed at concentration of 0.405% rosemary and 0.75 μg/mL nisin in both temprature. Results of this study showed that different concentration of rosemary a significant inhibitory effect (P<0.05) on log% of S. iniae, in BHI broth in pH 5.5 and 7 in 4,15 and 37 °C during 43 days. In concentration of 0% rosemary (control) in pH 5.5 and 7 and 37°C, log% were 1.099 and 3.15, whereas in concentration of 0.015% rosemary were -4/241 and 1.454, respectively. The use of essential oils may improve food safety and overall microbial quality. If essential oils were to be more widely applied as antibacterials in foods, the organoleptic impact would be important. In addition, it is recommended to apply essential oils or their compounds as part of a hurdle system and to use it as an antimicrobial component along with other preservation techniques. Thus essential of R. officinalis with high antibacterial activity selected in this study could be a potential source for inhibitory substances against some food-borne pathogens and they may be candidates for using in foods or food-processing systems.

A comparative study on effects of Immunoster and Immunowall as immunostimulants on some hematological, biochemical and growth indices of farmed great sturgeon juveniles (Huso huso)

Prebiotics are non-digestible food ingredients that profitably affect the host by selectively stimulating the growth and /or activation of one or a limited number of bacteria in the intestine that can enhance host health status. Immunoster (IS) and Immunowall (IW) are prebiotics and immunostimulants derived from the outer cell wall of brewers yeast, Saccharomyces cerevisiae. These substances contain MOS and �-glucans. After a four-week acclimatization period to rearing conditions and basal diet, 450 farmed great sturgeon juveniles weighing 95.58 ± 9.38 g were randomly distributed into 15 fiberglass tanks (2 × 2 × 0.53 m) in five treatments (Control, IS 1%, IW 1%, IS 3%, and IW 3%) in three replicates (completely randomized design) and kept at a density of 30 fish per tank for a period of 8 weeks at water temperature 20.55 ± 5.11ºC, dissolved oxygen 6.73 ± 0.35 mg L-1 and pH 7.92 ± 0.09. IS and IW were added at two levels of 1% and 3% to the basal diet in place of cellulose, except the control. At the beginning, in the middle and at the end of the trial, carcass analysis was done to determine the moisture, protein, fat, ash, and total carbohydrate. Also, blood samples were collected to measure hematological, biochemical and immune indices. At the end of the trial, final weight, final length, body weight increase (BWI), specific growth rate (SGR), average daily growth (ADG), protein efficiency ratio (PER), feed conversion ratio (FCR), and condition factor (CF) in fish fed on IS and IW in both levels 1% and 3% showed some differences. These differences were significant in IS 3% and IW 1% and 3% compared with the control (P<0.05). HSI showed no significant difference (P>0.05) and survival rate was 100% in all treatments. Crude protein of carcass in fish fed on IS and IW at 1% and 3% showed an increase in comparison with the control. There was significant difference between IS 3% and the control in crude protein of carcass (P<0.05). Fish fed on IS and IW at 1% and 3% showed various results in hematological and biochemical factors. It was observed significant difference in MCV between IW 1% and IS 3% compared with the control (P<0.05). Although there was an increase in values of hematocrit, hemoglobin (except IS 1%), WBC (except IW 3%), MCH, neutrophil, total protein, albumin (except IS 3%), K+, and lysozyme in fish fed on IS and IW compared with the control, it was no significant (P>0.05). The maximum count of WBC and the highest value of Ca2+ were seen in IW 1%. The maximum count of lymphocyte, the highest values of total protein, albumin and IgM were recorded in IW 3%. IS 1% had the maximum count of neutrophil and the highest concentration of lysozyme. Based on obtained results, it can be declared that IS and IW at two levels of 1% and 3% can enhance growth performance and feed efficiency and also improve some hematological, biochemical, and immune indices in farmed great sturgeon juveniles.

Growth and antioxidant system of the cyanobacterium Synechococcus elongatus in response to microcystin-RR

Microcystins, one type of the cyanobacterial toxins, show a broad range of hazardous effects on other organisms. Most of the researches on the toxic effects of microcystins have involved in animals and higher plants. Little work, however, has been done on evaluating the mechanisms of microcystin toxicity on algae. In this study, the toxicological effects of microcystin-RR (MC-RR) on the cyanobacterium Synechococcus elongatus were investigated. For this purpose, six physio-biochemical parameters (cell optical density, reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-Px) and glutathione S-transferase (GST)) were tested in algal cells when exposed to 100 mug(-1) microcystin-RR. The results showed that the growth of Synechococcus elongatus ( expressed as optical density) was significantly inhibited compared with the control. At the same time, the treated algae exhibited a pronounced increase in production of ROS and MDA after 6 days exposure to microcystin-RR. Signi. cant changes in GSH levels and GSH-Px, GSH activities were also detected in algal cells, with higher values being observed in the toxin treated algae after 6 days exposure. GST activities in the treated algae exhibited a decline after exposure and rapid augmentation on day 3, thereafter, they kept at a high level when compared to the control group. GSH contents and GSH-Px activities were also significantly raised in the toxin-treated algae cells from day 3, but they showed a sharp decrease on day 4, which was the onward of cell proliferation. These results suggested that oxidative stress manifested by elevated ROS levels and MDA contents might be responsible for the toxicity of microcystin to Synechococcus elongatus and the algal cells could improve their antioxidant ability through the enhancement of enzymatic and non-enzymatic preventive substances.