A Perda Auditiva Induzida pelo Ruído (PAIR) na formação odontológica: conhecimentos e níveis de exposição

The rationalization of work in the Dentistry has been taking the professional to work for ways and systems based in the ergonomics, turning their work efficient and less tiring. Since their academic formation, the dentists surgeons are concerned with the high productivity in clinic and with the final result of the work, neglecting the way as it is executed, which reduce their work capacity and exhibits them to occupational diseases that could be minimized and/or forewarned. This research had as the main objective to investigate the knowledge of the Dentistry academics of Rio Grande do Norte Federal University concerning the Noise-induced Hearing Loss (NIHL), relating them at the noise levels that they are exposed, as well as to the preventive measures taken during the clinical activities. Was observed that 95% of the individuals know that the dentist surgeon is a professional in risk for NIHL. Among the causes of NIHL, the one that obtained the largest frequency citation was the high-speed handpieces, reminded by 92,4% of the academics. Among the students which enumerated protective measures for NIHL, 92% mentioned the use of the ear plugs, although 97% of the researched have told do not use any kind of preventive measure related to the noise. Was also observed that 96% of the academics notice the noise during the clinical attendance, what inconvenience 28,1% of them. Related the noise levels, the high-speed handpieces of the academics presented a medium value of 80,5 dB varying from 72,3 to 88,3 dB. The average of the ambient noise observed at the Integrated Clinic was about 74,8 dB. In spite of the noise levels in this research were observed below the established limits of tolerance by the legislation, they can provoke damages to the Dentistry professionals' health, or that suggests the need of an intervention and use of immediate preventive measures able to generate a healthy atmosphere of work and less risky

Avaliação do efeito anti-inflamatório dos fármacos: atorvastatin, carvedilol, olmesartan e telmisartan, em modelo experimental da doença periodontal induzida em ratos

A periodontite é uma doença crônica inflamatória mediada por marcadores inflamatórios, tais como as citocinas: IL-1β, IL-10 e TNF-α, que provoca a destruição dos tecidos gengivais e osso alveolar, causando perda de inserção dentária e posterior perda dental. A perda óssea é causada pela ativação de prostaglandinas oriundas do ácido araquidônico, através da ação da enzima ciclooxigenase 2 (COX-2), promovendo a liberação de enzimas proteolíticas, as metaloproteinases de matriz, principalmente a MMP-2 e MMP-9, que promovem reabsorção óssea. Além disso, ocorre o desequilíbrio entre a ação de RANKL e OPG, havendo uma maior ativação de RANKL, e por consequência a maior ativação de osteoclastos e maior reabsorção óssea. Mediadores inflamatórios e espécies reativas de oxigênio (ROS) produzidos localmente possuem potencial para disseminar na corrente sanguínea e iniciar ou exacerbar doenças sistêmicas como as cardiovasculares. O tratamento atual da doença consiste em terapêutica local, mas a necessidade de estudos sobre fármacos de atuação sistêmica culminou nesta pesquisa, que realizou a avaliação dos fármacos: atorvastatin, carvedilol, olmesartan e telmisartan, quanto a sua ação anti-inflamatória sobre a doença periodontal induzida por ligadura em ratos Wistar. Os animais foram divididos em 5 grupos, para cada fármaco, separadamente: (NL) grupo não ligado, (L) grupo ligado sem tratamento, (1mg/Kg) grupo ligado que recebeu dose de 1mg/Kg de fármaco, (5 ou 6 mg/Kg) grupo ligado que recebeu dose de 5 ou 6 mg/Kg de fármaco, (10 mg/Kg) grupo ligado que recebeu dose de 10mg/Kg de fármaco. Foram realizadas avaliações: histopatológica, perda óssea alveolar, imuno-histoquímica (para COX-2, MMP-2, MMP-9, RANK-L, RANK e OPG), e ELISA (para mieloperoxidase, glutationa, malonaldeído e as citocinas: IL-1β, IL-10 e TNF-α). Os grupos tratados com olmesartana a 6 mg/Kg, e atorvastatin, carvedilol e telmisartan a 10mg/Kg, mostraram diminuição da perda óssea, redução de: MPO, MDA, IL-1β, TNF-α, MMP-2, MMP-9, COX-2, RANKL/RANK, e aumento na expressão da OPG e da IL-10.

Incidência de cárie dentária em adolescentes em município do Nordeste brasileiro, 2006

NORO, Luiz Roberto Augusto et al. Incidência de cárie dentária em adolescentes em município do Nordeste brasileiro, 2006, Cadernos de Saúde Pública, Rio de Janeiro, v. 25, n. 4, p. 783-790, abr. 2009.

Pré-condicionamento isquêmico em diferentes tempos e seu efeito na translocação bacteriana induzida por isquemia e reperfusão intestinal em ratos

Para avaliar os efeitos de diferentes tempos de pré-condicionamento isquêmico (IPC) em translocação bacteriana intestinal (BT). MÉTODOS: Trinta ratos Wistar pesando 280 ± 27g foram divididos em cinco grupos. No grupo IV (n = 6), a laparotomia foi realizada e a artéria mesentérica superior foi obstruído por um microclampe atraumática durante 30 minutos. Nos quatro grupos de pré-condicionamento (n = 6 cada) antes dos 30 minutos de isquemia-reperfusão (I / R), os ratos foram submetidos a IPC para duas, cinco, dez e 15 minutos, seguido pelo mesmo momento da reperfusão. A fim de avaliar se o tempo de pré-condicionamento influenciaram o surgimento de translocação bacteriana, as amostras de nódulos linfáticos mesentéricos, fígado e baço foram colhidas em condições estéreis, 24 horas após os procedimentos para a quantificação de unidades formadoras de colónias de bactérias por grama de tecido (CFU / g). O sangue foi recolhido para a medição de citoquinas. RESULTADOS: No grupo I / R, o total de CFU / g em gânglios linfáticos mesentéricos, baço, fígado, bem como o soro de TNF-a, IL-1A e IL-6 foram significativamente mais elevados do que nos outros grupos (p <0,05). Pré-condicionamento por 15 minutos significativamente atenuada BT e citocinas séricas quando comparado a outros períodos de pré-condicionamento (p <0,05). CONCLUSÃO: Nossos dados sugerem que o pré-condicionamento como um fator chave para reduzir a translocação bacteriana intestinal em I / R. Numa escala de dois a 15 minutos, o melhor tempo de pré-condicionamento isquémico pela atenuação da translocação bacteriana foi de 15 minutos

Efeito de glucanas do fungo Caripia montagnei em modelo de inflamação intestinal induzida por tnbs em ratos Wistar e em células de carcinoma de cólon humano HT-29

Compounds derived from fungi has been the subject of many studies in order to broaden the knowledge of their bioactive potential. Polysaccharides from Caripia montagnei have been described to possess anti-inflammatory and antioxidant properties. In this study, glucans extracted from Caripia montagnei mushroom were chemically characterized and their effects evaluated at different doses and intervals of treatment. It was also described their action on colonic injury in the model of colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS), and its action on cells of the human colon carcinoma (HT-29). Compounds extracted of C. montagnei contain high level of carbohydrates (96%), low content of phenolic compounds (1.5%) and low contamination with proteins (2.5%). The (FT-IR) and (NMR) analysis showed that polysaccharides from this species of mushroom are composed of α- and β-glucans. The colonic damage was evaluated by macroscopic, histological, biochemical and immunologic analyses. The results showed a reduction of colonic lesions in all groups treated with the glucans of Caripia montagnei (GCM). GCM significantly reduced the levels of IL-6 (50 and 75 mg/kg, p < 0.05), a major inflammatory cytokine. Biochemical analyses showed that such glucans acted on reducing levels of alkaline phosphatase (75 mg/kg, p < 0.01), nitric oxide (p < 0.001), and myeloperoxidase (p < 0.001). These results were confirmed microscopically by the reduction of cellular infiltration. The increase of catalase activity suggest a protective effect of GCM on colonic tissue, confirming their anti-inflammatory potential. GCM displayed cytostatic activity against HT-29 cells, causing accumulation of cells in G1 phase, blocking the cycle cell progression. Those glucans also showed ability to modulate the adhesion of HT-29 cells to Matrigel® and reduced the oxidative stress. The antiproliferative activity against HT-29 cells displayed by GCM (p <0.001) can be attributed to its cytostatic activity and induction of apoptosis by GCM

Efeito de frações obtidas da fucoidana de Fucus vesiculosus em modelo experimental de artrite induzida por Zymosan

The fucoidan from Fucus vesiculosus is known for having diverse biological properties. This study analyzed the therapeutic action of populations of commercial fucoidan (F. vesiculosus) on zymosan-induced arthritis. Three populations of fucoidan were obtained after acetone fractionation; these were denominated F1 (52.3%), F2 (36.7%) and F3 (10.7%). Chemical analyses showed that F1 contained the largest amount of sulfate ion. The electrophoretic profile shows that the commercial or total fucoidan (TF), different from the other fucoidans and from glycosaminoglycan patterns, is quite polydisperse, which indicates that it is composed of a mixture of sulfate polysaccharides. On the other hand, the fucoidans obtained from TF showed only an electrophoretic band with much lower polydispersion than that observed for TF. Fucoidan F2 showed a migration between fucoidans F1 and F3. Owing to the small amount of mass obtained from F3, we used only fucoidans F1 and F2 in the induced arthritis tests. After 1 hour of induction, we administered F1 or F2 (10, 25 and 50 mg/kg i.p.) or diclofenac sodium (10 mg/kg i.p.) or lumiracoxib (5 mg/kg o.a.) or L-NAME (30 mg/kg i.p.). After 6 hours, we performed analyses of cell influx and nitrite levels in addition to histopathological analysis. Fucoidans F1 and F2 were more potent both in decreasing the number of leukocytes and the amount of nitric oxide found in the synovial fluid. This indicates that the anti-inflammatory mechanism of these fucoidans is not only related to selectin block, but also to nitric oxide synthesis inhibition

Mutagênese induzida por flavonóides presentes do decocto das cascas da aroeira (Schinus terebinthifolius, Raddi)

The decoction of Brazilian pepper tree barks (Schinus terebinthifolius, Raddi), is used in medicine as wound healing and antiinflamatory. Once extracts from this plant are used for acceleration of scar s process, it is important to study their mutagenic and genotoxic potential. In previous works in our laboratory, it was observed mutagenicity caused by the decoction when in high concentrations. Among the chemical compounds of this plant that could be able to induce mutation, the flavonoids were the only group that was referred to have either an oxidant or antioxidant potential. The flavonoids were isolated, purified and quantified by adsorptive column chromatography under silica gel, bacterial and in vitro genotoxic tests were realized to determine if the flavonoids were the responsible agents for this mutagenicity found. The tests realized with plasmidial DNA were indicative that the flavonoids are probably genotoxic, due to the presence of correlation between increase of the flavonoid concentration and in plasmidial DNA double strand breakage visualized in agarose gel, as well as they were capable to generated abasic sites shown by the in vitro treatment with exonuclease III. The same tests with plasmidial DNA in the presence of copper [10 µM] and of a Tris-HCl pH 7.5 [10 µM] buffer were realized with the isolated flavonoids to determine if there would be or not participation of reactive oxygen species (ROS). The transformation of plasmidial DNA in different bacterial strains proficient and deficient in DNA repair enzymes in the presence or not of a Tris-HCl buffer, suggests that the enzymes that repair oxidative lesions are necessary to repair the lesions generated by the flavonoids and that ROS are generated and are necessary to promote the lesions. Bacterial tests with Escherichia coli strains of the CC collection (deficient or not for DNA repair enzymes), showed that the flavonoids are able to increase the frequency of mutations, mainly in strains mutated in repair enzymes (MutM, MutY-glicosylases and double mutant), suggesting that these agents are responsible for the enhancement in the mutation rate. In order to determine the mutation spectrum caused by the flavonoids of the Brazilian pepper tree stem bark, plasmidial DNA previously treated with the flavonoids were transformed in bacterial strains deficient and proficient in the DNA repair enzymes, followed by a blue-white selection with X-gal, DNA amplification by PCR and sequencing the positive mutant clones. Analysis of the mutants obtained from strains CC104, CC104mutM, CC104mutY, CC104mutMmutY, BW9101, BW9109 indicated a predominance of some mutations like G:C to C:G that can be correlated with the origin of 8-oxoG, due to oxidative lesions caused by the flavonoids. So it can concluded that the flavonoid isolated or in fractions enriched on them are genotoxic and mutagenic, and their mutations are predominantly oxidative, mediated by ROS, and the lesions are recognized by the BER system. In this way it is proposed that the flavonoids can act in two different ways to generate the DNA lesion: 1. in a Fenton-like reaction, when the flavonoid are in the presence of metal ions and that together with the water generate ROS that promotes the DNA lesions; 2. in another way the lesions can be generated by the formation of ROS due to the internal chemical structure of the flavonoid molecule due to the quantity and location of hydroxyl groups, and so producing the DNA lesions, those lesions can be directly (suggested by the in vitro experiments) or indirectly done (supported by the experiments using the CC bacterial strains)

Análise de parâmetros da função reprodutiva masculina após perda ponderal induzida pela cirurgia bariátrica

Introdução: A obesidade e suas formas extremas como a forma mórbida, vem assumindo preocupação nos sistemas de saúde pública e é atualmente considerado um dos mais importantes agravos de saúde no mundo desenvolvido. Acarreta repercussões em várias partes do corpo, com potencial para interferir negativamente também na função reprodutiva, recentemente sendo mais estudado na população masculina. Objetivo: avaliar parâmetros hormonais e seminais da função reprodutiva em um grupo de homens obesos mórbidos, antes e após 6 meses da perda de peso induzida pela cirurgia bariátrica. Métodos: Inicialmente, de uma amostra de 36 pacientes com idade entre 18 e 40 anos e índice de massa corporal (IMC) ≥40 Kg/m2, apenas 1 caso foi excluído por ser portador de hiperprolactinoma e outros 24 pacientes não completaram todo o protocolo de avaliação laboratorial solicitada. Um total de apenas 11 pacientes completou o protocolo. Todos foram previamente avaliados por uma equipe multidisciplinar. O grupo selecionado foi submetido a avaliação seminal e dosagens hormonais de testosterona total(TT), FSH e LH. Os parâmetros foram reavaliados após 6 meses de realização da cirurgia pela técnica de Y-em-Roux, para análise dos efeitos da perda de peso sobre os parâmetros da função reprodutiva. Resultados: No grupo selecionado ao estudo, os exames pré-operatórios mostraram um grupo heterogêneo de resultados. Após seis meses do tratamento cirúrgico, a perda ponderal foi significativa no grupo analisado, com redução do IMC de mediana 46 Kg/m2para mediana de 36 Kg/m2(p=0,03). Em relação ao perfil hormonal, houve elevação significativa do nível sérico de TT com mediana de 187ng/ml no período pré-operatório para 457 ng/ml (p=0,02) no período pós-operatório. Entre os períodos pré e pós-operatório, não foram observadas alterações significantes do FSH (2,70 mIU/ml para 3,31 mIU/ml, p=0,79), LH (4,82 mIU/ml para 3,00mIU/ml, p=0,39), concentração espermática (38milhões/ml para 31milhões/ml, p=0,88) e motilidade A+B (50% para 50%, p=0,89).Conclusão: A redução ponderal decorrente da cirurgia em obesos mórbidos parece levar a uma elevação dos níveis de testosterona total, sem alterações nas demais provas de avaliação reprodutiva. Estudos adicionais com casuística maior e múltiplos parâmetros são necessários para esclarecimento definitivo do impacto da obesidade e da perda de peso induzida pela cirurgia sobre a função reprodutiva masculina

Posicionamento e movimentação de um robô humanóide utilizando imagens de uma câmera móvel externa

This work proposes a method to localize a simple humanoid robot, without embedded sensors, using images taken from an extern camera and image processing techniques. Once the robot is localized relative to the camera, supposing we know the position of the camera relative to the world, we can compute the position of the robot relative to the world. To make the camera move in the work space, we will use another mobile robot with wheels, which has a precise locating system, and will place the camera on it. Once the humanoid is localized in the work space, we can take the necessary actions to move it. Simultaneously, we will move the camera robot, so it will take good images of the humanoid. The mainly contributions of this work are: the idea of using another mobile robot to aid the navigation of a humanoid robot without and advanced embedded electronics; chosing of the intrinsic and extrinsic calibration methods appropriated to the task, especially in the real time part; and the collaborative algorithm of simultaneous navigation of the robots

Modelagem e simulação de um separador bifásico com inversão de fases induzida

The present work has the main goal to study the modeling and simulation of a biphasic separator with induced phase inversion, the MDIF, with the utilization of the finite differences method for the resolution of the partial differencial equations which describe the transport of contaminant s mass fraction inside the equipment s settling chamber. With this aim, was developed the deterministic differential model AMADDA, wich was admensionalizated and then semidiscretizated with the method of lines. The integration of the resultant system of ordinary differential equations was realized by means of a modified algorithm of the Adam-Bashfort- Moulton method, and the sthocastic optimization routine of Basin-Hopping was used in the model s parameter estimation procedure . With the aim to establish a comparative referential for the results obtained with the model AMADDA, were used experimental data presented in previous works of the MDIF s research group. The experimental data and those obtained with the model was assessed regarding its normality by means of the Shapiro-Wilk s test, and validated against the experimental results with the Student s t test and the Kruskal-Wallis s test, depending on the result. The results showed satisfactory performance of the model AMADDA in the evaluation of the MDIF s separation efficiency, being possible to determinate that at 1% significance level the calculated results are equivalent to those determinated experimentally in the reference works

Plasticidade induzida por treinamento locomotor na medula espinal intacta em ratos: correlatos morfológicos

The locomotion is one of the most important capabilities developed by the animals, whose improvement is dependent on several neural centers, including the spinal cord. This activity promotes a lot of spinal modifications that enable it to adapt and improve their connections. This study aimed to observe the morphological changes occurring in the spinal cord after locomotor training in intact rats. For that we used male Wistar rats, which were submitted to locomotor training in wheel activity in protocols 1, 3 and 7 days (30min/day), and the results were compared to a control group not subjected to exercise. Coronal sections of 40 μm of the lumbosacral spinal cord were subjected to immunohistochemical techniques anti-Egr1, anti-NMDA and anti-SP, to characterize the spinal plasticity related to these substances. Egr1-immunoreactive cells were increased in all laminas, essentially in those more intensely activated by locomotion, laminas IV-X levels L4-S3. All observed sections expressed NMDA-immunoreactivity. Analysis of SP in the spinal dorsal horn resulted no significant variations of this neuropeptide related to locomotion. The results suggest that locomotor training provides synaptic plasticity similar to LTP in all laminas of the lumbosacral spinal cord, in different intensities. However, the SP appears do not participate of this process in the spinal dorsal horn. This work will contribute for consolidating and characterization of synaptic plasticity in the spinal cord

Efeitos da perda de peso induzida pela cirurgia bariátrica sobre a função respiratória

Background: Obesity leads to alteration of lung volumes and capacities due to accumulation of fat in the chest wall and abdomen. Few studies have shown that weight loss induced by surgery improves lung function. Our objective was to evaluate the anthropometric development, pulmonary function, respiratory muscle, strength and endurance after weight loss induced by bariatric surgery. Methods: We evaluated in pre and post operative period variables of weight, BMI, NC, WHR and spirometric and respiratory pressure. Results: 39 subjects were evaluated, with age mean 35.9 ± 10.9 years, predominantly by women (76.3%). The weight mean decreased from 124.8 ± 17.5 kg to 88.8 ± 14.28 kg in post operative. The mean BMI ranged from 47,9 ± 5,6 Kg/m² to 34,3 ± 4,75 Kg/m². There was a significant increase in FVC from 3,63 ± 0,94 to 4,01±1,03, FEV1 from 3,03 ± 0,72 to 3,39 ± 0,85, FEF 25-75% from 3,41 ± 0,72 to 3,82 ± 0,94, PEF from 6,56 ± 1,47 to 7,81 ± 1,69, ERV from 0,35 ± 0,39 to 0,66 ± 0,38, MVV ranged from 103,43 ± 22,21 to 137,27 ± 29,84, all of them to p<0,01. The MIP and MEP showed no significant difference in pre and post operative. It was noted that for every centimeter reduced in neck circumference, an increase of 0.06 in FVC and 5.98 in MVV is observed. This is also observed in weight and BMI. Conclusion: We conclude that weight loss induced by bariatric surgery in obese provides a significant improvement in lung function and reduction of fat around the neck is more important in the generation of lung volume than the reduction of BMI

Efeito do tempo de repouso após fadiga induzida no desempenho neuromuscular do quadríceps femoral: ensaio clínico, controlado, randomizado, cego

Objective: To evaluate the effect of different resting periods, after induced muscle fatigue, in the quadriceps femoris neuromuscular performance, in healthy subjects. Methods: Sixty-four volunteers, of both genders, with an average of 21,8 ± 1,7 years and mean body mass index of 24,2 ± 3,7 Kg/m2 were randomly assigned into 4 groups: control group (was not induced fatigue); Experimental Group 1 (Exp. 1 1 minute of rest after fatigue); Experimental Group 3 (Exp. 3 3 minutes of rest after fatigue) and Experimental Group 5 (Exp. 5 5 minutes of rest after fatigue). The subjects were evaluated to the knee´s joint position sense (JPS), followed by 5 flexion-extension knee concentric isokinetic contractions at 60°/s, with concomitant recording of median frequency (Fmed) of rectus femoris (RF), vastus lateralis (VL) and vastus medialis (VM). Then they underwent a muscle fatigue protocol (30 flexion-extension knee concentric contractions at 60°/s) and were reevaluated on the isokinetic performance, Fmed and JPS. Blood lactate levels were measured before initial assessment, immediately after the fatigue protocol and 5 minutes after the end of the rest period. Results: The adoption of 3 minutes of rest was sufficient to restore the initial conditions for the peak torque normalized by body weight and the VL and VM Fmed. The joint position sense returned to its initial values with 1 minute rest. The lactate concentration remained high regardless of the adopted rest period. Conclusion: The use of 3-minute rest period is sufficient for the reestablishment of the neuromuscular parameters to the pre fatigue values.

Efeitos imediatos da laserterapia sobre o desempenho neuromuscular após fadiga muscular induzida: ensaio clínico controlado, randomizado e cego

The aim of this study was to investigate the immediate effects of laser therapy on neuromuscular performance in healthy subjects after a muscle fatigue. This is a clinical trial, controlled, randomized, blinded, attended by 80 volunteers of both genders, healthy, with ages between 18 to 28 years. Initially the volunteers performed an initial evaluation (EV1) using electromyography in the biceps muscle, associated with assessment in isokinetic dynamometry with 5 concentric contractions (60 °/s) for elbow flexion. The subjects were randomly allocated into 4 groups: G1 (control, n = 20), G2 (placebo, n = 20), G3 (pre-fatigue laser, n = 20), and G4 (post fatigue laser, n = 20). The muscular fatigue protocol had 30 concentric isokinetic contractions (120 °/s). We used a 808 nm laser, power of 100 mW, applied at the belly of the biceps muscle. After the speeches the volunteers performed a final evaluation (EV2). Test was applied to two-way ANOVA with post hoc Turkey, with a significance level of 5%. There was no significant difference in electromyographic evaluation. In dynamometric evaluation showed a drop in peak torque, peak torque normalized to body weight (p <0.001) and average power (p <0, 05) between the initial and final evaluations in control. Among the groups there was a significant difference between the control and the other groups in relation to peak torque (p <0.05), peak torque to body weight (p <0.001) and average power (p <0.05). Therefore, the low intensity laser therapy does not alter the immediate neuromuscular performance after fatigue

Avaliação da eficácia de selante ionomérico modificado por resina e escovação dentária na preservação de cárie oclusal

The purpose of this study was to evaluate the effectiveness during a twelve-month period, of resin-modified glass ionomer (Vitremer@) used as fissure sealants compared to supervised teeth brushing in the prevention of oclusal caries in permarient molars of pubIlc school children from 5 to 7 years of age. A total of 91 children participated in the study, being randomly divided up into three groups of investigation: group with sealant (n=31), group of supervised brushing (m=30), and control gorup (n=30). After 12 months, a total of 28.5% of tosses in the sample of 26 children was registered, and although there wasn t a significant diference between these losses and the groups studles (p = 0.6355), there was a considerable reduction in the sample. The results showed that none of the independentIy studied variables (present caries experience, biofilm, bleeding, sealant retention, position of the tooth in the arch, and sex) interfered in the final results, and that there was no significant difference between applying the sealant in question, perfornling supervised dental brushing, or not intervening at alI (p=0.542). These results could have been a consequence of the limitations found alI through the study, especially relating to the losses occurred because of school evasion, transfer, or because of the accident in the Marise Paiva Municipal School, Keeping students away for approximately one semester, so tht a new reseach had to be developed sos as to better clarify the effects among the treatments done there