Depression is a psychiatric disorder that affects 2-15% of the world population. It is a heterogeneous neuropathology, it includes psychological, behavioral and physiological symptoms. Despite having several treatments for depression, they are not as effective and have many undesirable side effects. Therefore, it is necessary the development of new therapeutic strategies to improve current treatments. Therefore, open up doors to the use of plant medicines, which has already been used for thousands of years by various populations for different diseases. The Bauhinia variegata, popularly known as "pata-de-vaca", has beneficial health effects, such as anti-inflammatory activity, antidiabetic, antidiarrheal. In the present study, we investigated the antidepressant-like effect of the crude hydroethanolic extract of leaves of Bauhinia variegata in two predictive tests of antidepressant property, the tail suspension test (TST) and the forced swimming test (FST) in mice. Furthermore, the involvement of monoaminergic system in its antidepressant-like effect was investigated and the effect of extract on monoamine oxidase A (MAO-A) activity was also evaluated. The acute treatment with the extract of Bauhinia variegate (300 mg/kg, p.o.) significantly reduced the immobility time in the TST. Moreover, the repeated administration (once a day for 7 days) of the extract by p.o. route also produced an antidepressant-like effect in the TST (100 and 300 mg/kg) and FST (300 mg/kg).The pretreatment of mice with ketanserin (5 mg/kg, i.p. a preferential 5-HT2A receptor antagonist), prazosin (1 mg/kg, i.p. an α1-adrenoceptor antagonist), yohimbine (1 mg/kg, i.p. an α2-adrenoceptor antagonist), propranolol (2 mg/kg, i.p. a β-adrenoceptor antagonist), SCH23390 (0.05 mg/kg, i.p. a dopamine D1 receptor antagonist) or sulpiride (50 mg/kg, i.p. a dopamine D2 receptor antagonist) was able to reverse the anti-immobility effect of the extract (300 mg/kg, p.o) in TST. The combination of subeffective dose of fluoxetine (5 mg/kg, p.o.), imipramine (5 mg/kg, v.o.) or bupropion (5 mg/kg, v.o.) with a subffective dose of the extract (100 mg/kg) produced a synergistic antidepressant-like effect in the TST. The extract did not cause motor alteration in the open-field test at effective doses in both TST and FST. We also showed that the extract inhibited the MAO-A activity of mice brain in vitro. These results suggest that the extract from Bauhinia variegata present an antidepressanloke potential, which may be dependent on the monoaminergic system and its MAO-A inhibitory properties. / A depressão é um transtorno psiquiátrico que atinge 2-15% da população mundial. É uma neuropatologia heterogênea, pois inclui sintomas psicológicos, comportamentais e fisiológicos. Apesar de possuir diversos tratamentos para depressão, eles ainda não são tão efetivos e possuem muitos efeitos colaterais indesejáveis. Por isso, faz-se necessário o desenvolvimento de novas estratégias terapêuticas a fim de aperfeiçoar os tratamentos atuais. Diante disso, abrem-se possibilidades para o uso de plantas medicinas, uma vez que estas vem sendo utilizadas há milhares de anos por muitas populações para tratar diversas enfermidades. A Bauhinia variegata, conhecida popularmente como pata-de-vaca , possui efeitos benéficos à saúde, como: atividade anti-inflamatória, antidiabética, antidiarréica, entre outras. No presente estudo, foi investigado o efeito tipo-antidepressivo do extrato hidroetanólico bruto de folhas de Bauhinia variegata em dois testes preditivos de propriedade antidepressiva, o teste de suspensão pela cauda (TSC) e do teste do nado forçado (TNF) em camundongos machos adultos. Além disso, foi investigado o envolvimento do sistema monoaminérgico no efeito do extrato e sua atividade sobre a monoamina-oxidase A (MAO-A). O extrato bruto hidroetanólico foi administrado por via oral (vo) por gavagem. O tratamento agudo com o extrato de Bauhinia variegata (300 mg/kg) reduziu o tempo de imobilidade no TST. Além disso, a administração repetida (uma vez por dia durante 7 dias) do extrato também produziu um efeito tipo-antidepressivo no TSC (100 e 300 mg/kg) e TNF (300 mg/kg). O pré-tratamento de camundongos com cetanserina (5 mg/kg, ip, antagonista do receptor serotoninérgico 5-HT2A), prazosina (1 mg/kg, ip, antagonista α1-adrenérgico), ioimbina (1 mg/kg, ip, antagonista α2-adrenérgico), propranolol (2 mg/kg, ip, antagonista β-adrenérgico), SCH23390 (0,05 mg/kg, ip, antagonista do receptor dopaminérgico D1) ou sulpirida (50 mg/kg, ip, antagonista do receptor dopaminérgico D2) reverteu o efeito anti-imobilidade do extrato (300 mg/kg) no TSC. A combinação de doses sub-efetivas dos antidepressivos clássicos, fluoxetina (5 mg/kg, vo), imipramina (5 mg/kg vo) ou bupropiona (5 mg/kg, vo) com uma dose sub-efetiva do extrato (100 mg/kg) produziu um efeito antidepressivo sinérgico no TSC. O extrato não causou alteração locomotora no teste de campo aberto em doses eficazes, tanto no TSC como no TNF. Além disso, o extrato inibiu a atividade da MAO-A em preparações de mitocôndria encefálicas in vitro. Estes resultados sugerem que o extrato hidroetanólico bruto de folhas de Bauhinia variegata apresenta um potencial efeito tipo-antidepressivo, que pode ser dependente do sistema monoaminérgico.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Depression is a serious, recurrent and incapacitating psychiatric condition with a heavy social burden. The pharmacological approach to this disorder employs therapy with antidepressant drugs, which have side effects and numerous limitations. In view of the promising pharmacological properties of containing-selenium molecules, this study evaluated the effect of 3-(4-fluorophenylselenyl)-2,5-diphenylselenophene (DPS) in the mouse forced swim test (FST) and tail suspension test (TST), two models predictive of depressant activity. Since serotonin (5-HT) plays an important role in the pathophysiology of depressive disorders, the involvement of serotonergic system and 5-HT receptors in the action caused by DPS was studied. The antidepressant-like action of combined treatment with subeffective doses of both DPS plus paroxetine, a selective serotonin reuptake inhibitor (SSRI) was investigated. Further, we verified the possible mechanism responsible for antidepressant-like action of DPS. The results showed that DPS (50 and 100 mg/kg, p.o.) significantly reduced the immobility time during the FST and TST, without accompanying changes in ambulation when assessed in the open-field test. The anti-immobility effect of DPS (50 mg/kg, i.g.) in the FST was prevented by pretreatment of mice with pCPA (p-chlorophenylalanine; an inhibitor of 5-HT synthesis, 100 mg/kg, i.p., once a day for 4 consecutive days,), WAY 100635 (N-[2- [4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-2-pyridinylcyclohexane carboxamide; 0.1 mg/kg, s.c., a selective 5-HT1A receptor antagonist), ritanserin (1 mg/kg, i.p., a 5-HT2 receptor antagonist) or ondansetron (1 mg/kg, i.p., a 5-HT3 receptor antagonist). Combined treatment with paroxetine and DPS reduced the immobility time in the FST. DPS at the dose of 50 mg/kg did not produce any change in the cerebral activity of monoamine oxidase subtypes (MAO-A or MAO-B). DPS at the dose of 50 mg/kg inhibited significantly 5-HT uptake in mouse brain synaptosomes. These results suggest that DPS produced an antidepressant-like action in the mouse FST and TST and this action seems most likely to be mediated through an interaction with serotonergic system, particularly by 5-HT reuptake inhibition. / A depressão é uma doença grave, recorrente e uma condição psiquiátrica incapacitante que gera pesados custos sociais. A abordagem farmacológica dessa desordem é feita por meio do emprego de antidepressivos, os quais apresentam efeitos adversos e numerosas limitações. Tendo em vista as promissoras propriedades farmacológicas das moléculas contendo selênio, este estudo avaliou o efeito do 3-(4-fluorofenilselenil)-2,5-difenilselenofeno (DPS) no teste do nado forçado (TNF) e no teste da suspensão da cauda (TSC) em camundongos, dois modelos preditivos de comportamento depressivo. Uma vez que a serotonina (5-HT) desempenha um importante papel na patofisiologia dos transtornos depressivos, o envolvimento do sistema serotoninérgico e dos receptores de 5-HT na ação desenvolvida pelo DPS foi estudado. A ação do tipo antidepressiva do tratamento combinado com doses subefetivas de DPS e paroxetina, um inibidor seletivo da recaptação de serotonina (ISRS) foi investigada. Além disso, o possível mecanismo responsável pela ação do tipo antidepressiva do DPS também foi verificado. Os resultados mostram que o DPS (50 e 100 mg/kg) reduziu significativamente o tempo de imobilidade durante os TSC e TNF, sem causar alterações na atividade locomotora no teste do campo aberto (TCA). O efeito anti-imobilidade do DPS (50 mg/kg, i.g.) no TNF foi prevenido pelo pré-tratamento dos animais com pCPA (p-clorofenilalanina; 100 mg/kg, i.p., administrado uma vez ao dia durante 4 dias consecutivos, um inibidor da síntese de serotonina), WAY 100635 (N-[2-[4-(2-metoxifenil)-1-piperazinil]etil]-N-2- piridinilciclohexano carboxamida; 0,1 mg/kg, s.c., um antagonista seletivo de receptores 5- HT1A), ritanserina (1 mg/kg, i.p., um antagonista seletivo de receptores 5-HT2) ou ondansetrona (1 mg/kg, i.p., um antagonista seletivo de receptores 5-HT3). O tratamento combinado com paroxetina e DPS reduziu o tempo de imobilidade no TNF. O DPS na dose de 50 mg/kg não produziu nenhuma alteração na atividade dos subtipos da monoamino oxidase (MAO-A e MAO-B) cerebral. O DPS na dose de 50 mg/kg inibiu significantemente a captação de 5-HT em sinaptossoma de cérebro de camundongos. Esses resultados sugerem que o DPS produziu uma ação do tipo antidepressiva no TSC e no TNF em camundongos e esta ação parece ser mediada por uma interação com o sistema serotoninérgico, particularmente por uma inibição da recaptação de 5-HT.

Einführung: Enterobacter (E.) cloacae subsp. cloacae sind wichtige Humanpathogene, insbesondere bei stationär untergebrachten Patienten. Sie sind in der Lage, Medizinprodukte zu kontaminieren, und es wurde über nosokomiale Krankheitsausbrüche in Verbindung mit der Kolonisation von chirurgischen Utensilien berichtet. Es ist daher wichtig, die Wirksamkeit und Effizienz von Desinfektionsmitteln gegenüber dieser Bakterienart zu bestimmen. Ziele: Die aktuelle Studie wurde durchgeführt, um nachzuweisen, ob Peressigsäure, Ethanol, Benzalkoniumchlorid und Natriumhypochlorit, welche weit verbreitet in kommerziellen Desinfektionsmitteln enthalten sind, eine ausreichende Wirksamkeit gegen multiresistente, von Patienten im Krankenhaus isolierte, E. cloacae aufweisen. Material und Methoden: Sechs multiresistente E. cloacae Isolate, die von Patienten in einem klinischen Umfeld gewonnen wurden, wurden getestet und mit dem E. cloacae Typstamm verglichen. Die Studien wurden in vitro mit Peressigsäure, Ethanol, Benzalkoniumchlorid und Natriumhypochlorit nach den Richtlinien der Desinfektionsmittel-Kommission des Verbundes für Angewandte Hygiene e.V. durchgeführt. Die Tests umfassten qualitative und quantitative Suspensionstests zur Bestimmung der bakteriziden Wirkung, den sogenannten Keimträgertest und die Bestimmung der minimalen Hemmkonzentrationen. Ergebnisse: Die Studienergebnisse zeigten, dass multiresistente E. cloacae Stämme genauso empfindlich gegenüber Desinfektionsmitteln waren wie der Typstamm. Organische Belastung interagierte stark mit Natriumhypochlorit und minderte dadurch seine Wirksamkeit, während Peressigsäure und Ethanol nicht durch organische Verunreinigung beeinflusst wurden. Die Kontaktzeit hatte nur einen geringen Einfluss auf die bakterizide Wirkung. Im Gegensatz dazu spielten bei Benzalkoniumchlorid organische Verunreinigung und die Kontaktzeit eine wichtige Rolle. Insgesamt waren die minimalen Hemmkonzentrationen und die bakterizid wirksamen Konzentrationen niedriger als die für kommerzielle Produkte gebräuchlichen Konzentrationen. In den Keimträgertests hatte das Trocknen auf einer glatten Oberfläche einen Einfluss auf das Überleben eines Stammes von E. cloacae. Die Ergebnisse zeigten auch, dass sich die Wirksamkeit der Desinfektionsmittel in den verschiedenen verwendeten Tests deutlich unterscheiden kann. Die Ergebnisse waren schwer mit anderen Studien zu vergleichen, da eine internationale Durchführungsrichtlinie für die Prüfung der Wirksamkeit von Desinfektionsmitteln gegen multiresistente Bakterien fehlt. Fazit: Peressigsäure, Ethanol, Benzalkoniumchlorid und Natriumhypochlorit eignen sich zur Desinfektion von multiresistenten E. cloacae. Die Wirksamkeit von Natriumhypochlorit und Benzalkoniumchlorid wird jedoch stark durch organische Stoffe beeinflusst. Dies unterstreicht die Bedeutung geeigneter Reinigungsmaßnahmen vor der Desinfektion. Wenn dies erfolgt ist, erweisen sich die getesteten Desinfektionsmittel gegen multiresistente E. cloacae genauso effektiv wie gegen den Typstamm.:1. Introduction ............................................................ 1 2. Literature Review ....................................................... 3 2.1 Enterobacteriaceae ..................................................... 4 2.1.1 General properties ................................................... 4 2.1.2 Enterobacter cloacae complex ......................................... 4 2.2 Multidrug-resistant bacteria and disinfectant “resistance” ............. 6 2.3 Disinfectant testing ................................................... 7 2.4 Active substances investigated in this study ........................... 8 2.4.1 Peracetic acid (PAA) ................................................. 8 2.4.2 Ethanol (ETH) ........................................................ 9 2.4.3 Benzalkonium chloride (BKC) .......................................... 9 2.4.4 Sodium hypochlorite (NaOCl) .......................................... 10 3. Materials and Methods ................................................... 11 3.1 Materials .............................................................. 11 3.2 Methods ................................................................ 14 3.2.1 Culture and storage of bacteria ...................................... 14 3.2.2 Preparation of disinfectants and the neutralizing agent .............. 14 3.2.3 Minimum inhibitory concentration (MIC) ............................... 15 3.2.4 Qualitative suspension test .......................................... 15 3.2.5 Quantitative suspension test.......................................... 16 3.2.6 Surface disinfection without mechanical action (germ carrier test) ... 16 3.2.7 Statistical analysis ................................................. 17 4. Results ................................................................. 18 4.1 Minimum inhibitory concentrations ...................................... 18 4.2 Qualitative suspension tests ........................................... 18 4.3 Quantitative suspension tests .......................................... 21 4.4 Surface disinfection without mechanical action (germ carrier test) ..... 24 5. Discussion .............................................................. 28 6. Summary ................................................................. 31 7. Zusammenfassung ......................................................... 33 8. References .............................................................. 35 9. Appendix ................................................................ 49 Acknowledgments ............................................................ 50 / Introduction: Enterobacter (E.) cloacae subsp. cloacae are important human pathogens, particularly in hospitalized patients. They tend to contaminate various medical devices and nosocomial outbreaks have been reported to be associated with the colonization of surgical equipment. Therefore, it is critical to determine the efficacy and effectiveness of disinfectants against this bacterial species. Objectives: The current study was undertaken to prove whether single active ingredients (i.e. peracetic acid, ethanol, benzalkonium chloride, and sodium hypochlorite) of widely used commercial disinfectants provide proper efficacy against multidrug-resistant human isolates of E. cloacae. Material and Methods: Six multidrug-resistant E. cloacae isolates obtained from patients in a clinical setting were tested and compared to the E. cloacae type strain. The studies were performed in vitro using peracetic acid, ethanol, benzalkonium chloride and sodium hypochlorite following the guidelines specified by the Disinfectants Commission within the Association of Applied Hygiene. Tests included determination of minimum inhibitory concentrations, bactericidal values by qualitative and quantitative suspension tests, and so-called germ carrier tests. The influence of exposure time and organic load on bacteriostatic and bactericidal concentrations was evaluated for each disinfectant using the two-tailed Mann-Whitney U-test. Results: Study results showed that multidrug-resistant E. cloacae strains were equally susceptible to disinfectants as the type strain. Organic matter highly interfered with sodium hypochlorite thereby decreasing its efficacy whereas peracetic acid and ethanol were not influenced by organic soiling. Contact time had only a minor effect on bactericidal values. This was in contrast to benzalkonium chloride where organic soiling and contact time played an important role. On the whole, minimum inhibitory concentrations and bactericidal concentrations were lower than in-use concentrations of commercial products. Drying on smooth surfaces in the carrier tests had an effect on the survival of one E. cloacae strain. Results also showed that efficacious values determined by the different tests used may differ distinctly. Results were difficult to compare with other studies because an international practical standard for testing disinfectant efficacy against multidrug-resistant bacteria is missing. Conclusion: Peracetic acid, ethanol, benzalkonium chloride and sodium hypochlorite are suitable to disinfect multidrug-resistant E. cloacae but the effectiveness of sodium hypochlorite and benzalkonium chloride is strongly influenced by organic matter. This underlines the importance of proper cleaning measures before disinfection. When this is done, the tested disinfectants proved to be as efficient against multidrug-resistant E. cloacae as against the type strain.:1. Introduction ............................................................ 1 2. Literature Review ....................................................... 3 2.1 Enterobacteriaceae ..................................................... 4 2.1.1 General properties ................................................... 4 2.1.2 Enterobacter cloacae complex ......................................... 4 2.2 Multidrug-resistant bacteria and disinfectant “resistance” ............. 6 2.3 Disinfectant testing ................................................... 7 2.4 Active substances investigated in this study ........................... 8 2.4.1 Peracetic acid (PAA) ................................................. 8 2.4.2 Ethanol (ETH) ........................................................ 9 2.4.3 Benzalkonium chloride (BKC) .......................................... 9 2.4.4 Sodium hypochlorite (NaOCl) .......................................... 10 3. Materials and Methods ................................................... 11 3.1 Materials .............................................................. 11 3.2 Methods ................................................................ 14 3.2.1 Culture and storage of bacteria ...................................... 14 3.2.2 Preparation of disinfectants and the neutralizing agent .............. 14 3.2.3 Minimum inhibitory concentration (MIC) ............................... 15 3.2.4 Qualitative suspension test .......................................... 15 3.2.5 Quantitative suspension test.......................................... 16 3.2.6 Surface disinfection without mechanical action (germ carrier test) ... 16 3.2.7 Statistical analysis ................................................. 17 4. Results ................................................................. 18 4.1 Minimum inhibitory concentrations ...................................... 18 4.2 Qualitative suspension tests ........................................... 18 4.3 Quantitative suspension tests .......................................... 21 4.4 Surface disinfection without mechanical action (germ carrier test) ..... 24 5. Discussion .............................................................. 28 6. Summary ................................................................. 31 7. Zusammenfassung ......................................................... 33 8. References .............................................................. 35 9. Appendix ................................................................ 49 Acknowledgments ............................................................ 50