An adjunctive antidepressant nutraceutical combination in treating major depression: study protocol and clinical considerations

Current treatment for major depressive disorder (MDD), a prevalent and disabling mental illness, is inadequate, with two-thirds of people treated with first-line antidepressants not achieving remission. MDD is for many a chronic condition, often requiring multiple treatment attempts, thus development of additional interventions is urgently required. An emerging approach to improve non-response to antidepressants is the use of adjunctive nutraceuticals. The pathophysiology of MDD is considered to involve a range of abnormalities (monoamine impairment, neuro-endocrinological changes, reduced brain-derived neurotrophic factor, and cytokine alterations). By targeting an array of these key neurobiological pathways via specific nutraceuticals (S-adenosyl methionine; [SAMe], 5-HTP [active tryptophan], folinic acid [active folic acid], omega-3 fatty acids, and zinc), there is the potential to provide a more comprehensive therapeutic biological approach to treat depression. We are currently conducting a National Health and Medical Research Council funded study in Australia (APP1048222). The clinical trial is phase II/III, multi-site, 3-arm, 8-week, randomised, double-blind, placebo-controlled study using SAMe + folinic acid versus a combination nutraceutical (SAMe, 5-HTP, folinic acid, omega-3, and zinc) or matching placebo in 300 currently depressed participants with diagnosed MDD who are non-responsive to current antidepressants (ANZCTR, protocol number: 12613001300763). The results may provide evidence for a novel adjunctive neurobiological approach for treating depression.

Nutritional medicine as mainstream in psychiatry

Psychiatry is at an important juncture, with the current pharmacologically focused model having achieved modest benefits in addressing the burden of poor mental health worldwide. Although the determinants of mental health are complex, the emerging and compelling evidence for nutrition as a crucial factor in the high prevalence and incidence of mental disorders suggests that diet is as important to psychiatry as it is to cardiology, endocrinology, and gastroenterology. Evidence is steadily growing for the relation between dietary quality (and potential nutritional deficiencies) and mental health, and for the select use of nutrient-based supplements to address deficiencies, or as monotherapies or augmentation therapies. We present a viewpoint from an international collaboration of academics (members of the International Society for Nutritional Psychiatry Research), in which we provide a context and overview of the current evidence in this emerging field of research, and discuss the future direction. We advocate recognition of diet and nutrition as central determinants of both physical and mental health.

The Glutathione System: a new drug target in neuroimmune disorders

Glutathione (GSH) has a crucial role in cellular signaling and antioxidant defenses either by reacting directly with reactive oxygen or nitrogen species or by acting as an essential cofactor for GSH S-transferases and glutathione peroxidases. GSH acting in concert with its dependent enzymes, known as the glutathione system, is responsible for the detoxification of reactive oxygen and nitrogen species (ROS/RNS) and electrophiles produced by xenobiotics. Adequate levels of GSH are essential for the optimal functioning of the immune system in general and T cell activation and differentiation in particular. GSH is a ubiquitous regulator of the cell cycle per se. GSH also has crucial functions in the brain as an antioxidant, neuromodulator, neurotransmitter, and enabler of neuron survival. Depletion of GSH leads to exacerbation of damage by oxidative and nitrosative stress; hypernitrosylation; increased levels of proinflammatory mediators and inflammatory potential; dysfunctions of intracellular signaling networks, e.g., p53, nuclear factor-κB, and Janus kinases; decreased cell proliferation and DNA synthesis; inactivation of complex I of the electron transport chain; activation of cytochrome c and the apoptotic machinery; blockade of the methionine cycle; and compromised epigenetic regulation of gene expression. As such, GSH depletion has marked consequences for the homeostatic control of the immune system, oxidative and nitrosative stress (O&NS) pathways, regulation of energy production, and mitochondrial survival as well. GSH depletion and concomitant increase in O&NS and mitochondrial dysfunctions play a role in the pathophysiology of diverse neuroimmune disorders, including depression, myalgic encephalomyelitis/chronic fatigue syndrome and Parkinson’s disease, suggesting that depleted GSH is an integral part of these diseases. Therapeutical interventions that aim to increase GSH concentrations in vivo include N-acetyl cysteine; Nrf-2 activation via hyperbaric oxygen therapy; dimethyl fumarate; phytochemicals, including curcumin, resveratrol, and cinnamon; and folate supplementation.

Efeitos da mal nutrição protéica sobre o metabolismo da glicina em cerebelo de ratos durante o seu desenvolvimento

Malnutrition is a worldwide problem affecting millions of unborn and young children during the most vulnerable stages of brain development (1). All restriction of protein during the perinatal period of life can alter the development of mammalian fetus and have marked repercussions on development of the Central Nervous System (CNS). The brain is vulnerable to protein malnutrition with altered morphologic and biochemical maturation, leading to impaired functions. The focus of this study is to investigate [U-14C]glycine metabolism in malnourished rats submitted to pre- and postnatal protein deprivation (diet: 8% protein with addition and without addition of L-methionine) on glycine metabolism of rats (normonourished group: 25% protein). It was observed that protein malnutrition alters oxidation to CO2, conversion to lipids and protein synthesis from [U-14C]glycine in cerebellum of malnourished rats without addition of L-methionine on a diet at 7 and 21 days of postnatal life. Our results also indicate that protein malnutrition causes a retardation in the normally ordered progression of brain development, and the malnourished groups have smaller cells, reduction in cell numbers and smaller cerebellar weight comparing to the control group.

Use of novel biomarkers (Homocysteine, vitamin B6, B9 and B12) on the assessing the progression of cardiovascular disease

A large number of evidences correlate elevated levels of homocysteine (Hcys) with a higher cardiovascular diseases (CVDs) risk, especially, atherosclerosis. Similarly, abnormal low levels of the vitamins B6, B9 and B12 are associated to an instability in the methionine cycle with an over production of Hcys. Thus, biomedical sciences are looking forward for a cheaper, faster, precise and accurate analytical methodology to quantify these compounds in a suitable format for the clinical environment. Therefore the objective of this study was the development of a simple, inexpensive and appropriate methodology to use at the clinical level. To achieve this goal, a procedure integrating a digitally controlled (eVol®) microextraction by packed sorbent (MEPS) and an ultra performance liquid chromatography (UPLC) coupled to a photodiode array detector (PDA) was developed to identify and quantify Hcys vitamins B6, B9 and B12. Although different conditions were assayed, we were not able to combine Hcys with the vitamins in the same analytical procedure, and so we proceeded to the optimization of two methods differing only in the composition of the gradient of the mobile phase and the injected volume. It was found that MEPS did not bring any benefit to the quantification of the Hcys in the plasma. Therefore, we developed and validate an alternative method that uses the direct injection of treated plasma (reduced and precipitated). This same method was evaluated in terms of selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), matrix effect and precision (intra-and inter-day) and applied to the determination of Hcys in a group composed by patients presenting augmented CVD risk. Good results in terms of selectivity and linearity (R2> 0.9968) were obtained, being the values of LOD and LOQ 0.007 and 0.21 mol / L, respectively. The intra-day precision (1.23-3.32%), inter-day precision (5.43-6.99%) and the recovery rate (82.5 to 93.1%) of this method were satisfactory. The matrix effect (>120%) was, however, higher than we were waiting for. Using this methodology it was possible to determine the amount of Hcys in real plasma samples from individuals presenting augmented CVD risk. Regarding the methodology developed for vitamins, despite the optimization of the extraction technique and the chromatographic conditions, it was found that the levels usually present in plasma are far below the sensitivity we obtained. Therefore, further optimizations of the methodology developed are needed. As conclusion, part of the objectives of this study was achieved with the development of a quick, simple and cheaper method for the quantification of Hcys.

Inclusão de farinha de resíduos da indústria de filetagem de tilápias na alimentação de girinos de rã-touro (Rana Catesbeiana Shaw,1802)

The objective of this experiment was to evaluate the inclusion of meal of tilapia industrial waster in bullfrog tadpoles' diets (Rana Catesbeina Shaw, 1802). Two hundred fifty tadpoles with initial weight and length of 0,18 +/- 0,05g and 2,50 +/- 0,18 centimeters were used, respectively, they were used and distributed in 25 aquariums of 30 liter each in a complete randomized design with five treatments and five repetitions. The tadpoles were fed four times a day with diets containing 0, 5, 10 and 15% of inclusion of tilapia filleting and another diet with 0% of supplemented tilapia filleting with synthetic methionine, being supplied four times a day at ease. The experiment period was 30 days. The diets with tilapia filleting inclusion and methionine supplementation did not have influence in the final weight and length averages, feed survival and conversion of bullfrog tadpoles. It cam be used until 15% of tilapia filleting inclusion in the bullfrog tadpoles food. The synthetic methionine supplementation in diets for bullfrog tadpoles is not necessary in diets with levels over 0,471/4 of the total methionine.

Plant or fungal sequences? An alternative optimized PCR protocol to avoid ITS (nrDNA) misamplification

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

Estudo de polimorfismos em genes relacionados ao metabolismo do ácido fólico e sua associação com o desenvolvimento de fendas orais não-sindrômicas

The congenital facial clefts are characterized by incomplete formation of the structures that separate the oral and nasal cavity. It is known that several environmental and genetic factors are involved in its development, among these, polymorphisms associated with folic acid metabolism have been investigated. In this sense, the objective was to observe the frequency of polymorphisms C677T and A1298C methylenetetrahydrofolate reductase gene (MTHFR), methionine synthase A2756G of (MTR), A66G of methionine synthase reductase (MTRR) A80G and the reduced folate carrier (RFC1) in patients with non-syndromic oral clefts, trying to match them with their development. Methods: We studied 140 patients with non-syndromic oral clefts and their mothers and 175 control subjects with their mothers, who underwent a questionnaire to obtain family information. Were collecting blood for DNA extraction from patients and their mothers to identify the genotypes of both by PCRRFLP, in addition to carrying out the determination of glucose, AST, ALT and serum creatinine, folic acid and vitamin B12 Serum and plasma homocysteine, and the hemogram. Results: Most patients have cleft lip and palate (55.8%), followed by isolated cleft palate (24.2%) and cleft lip (20%). Regarding gender, 62% of patients were male and 48% female and, after subdivision of the type of screwdriver according to sex was found a prevalence of males in the cracks of the type lip and palate (69 %) and lip (69.2%) and in the case of cleft palate was a female predominance (59%). The average concentration of serum folate in the group of mothers of cleft patients was significantly lower (13.8 ± 2.4 ng / mL) compared with the group of mothers of control subjects (18.8 ± 3.4 ng / mL) This was also observed for the group of cleft children as compared to controls, the dosage of folic acid had a significant difference with values of 15.6 ± 0.6 (ng / mL) and 17.9 ± 0.6 (ng / mL), respectively. For the biochemical measurements of glucose, AST, ALT and creatinine were not statistically different, nor was observed for haematological parameters performed. In assessing the frequency of polymorphisms C677T and A1298C MTHFR, A2756G MTR, MTRR A66G and A80G of the RFC1 there was no statistically significant difference in genotype distribution between cases and controls both for mothers and in the cleft. Conclusion: Although not observed association of polymorphisms with the development of cracks, the decrease in serum folate in the group of cleft patients and their mothers may reflect a disturbance in the metabolism of this metabolite, necessitating further studies such as studies methylation and expression to further elucidate the involvement of folate in the development of oral clefts

Efeito da metionina protegida e não protegida da degradação ruminal sobre a produção e composição do leite de vacas holandesas

Utilizaram-se vacas Holandesas com produção superior a 20 kg de leite/dia, de primeira e segunda lactações, com 19±6 dias em lactação, para avaliar o efeito da suplementação com 8,4 g/dia de metionina protegida (MPDR) ou 8,4 g/dia de metionina não-protegida da degradação ruminal (MNPDR) sobre a produção e composição do leite, comparativamente a vacas controle, durante 90 dias. As vacas foram alimentadas com ração completa constituída por silagem de milho e concentrado. Produção de leite, teor de proteína do leite e produção de proteína não foram afetados pela suplementação com MPDR. As produções médias de leite foram 27,70; 27,09 e 27,61 kg/dia; os teores médios de proteína, 2,83; 2,85 e 2,77%; e as produções de proteína do leite, 0,77; 0,76 e 0,79 kg/dia, respectivamente, para vacas controle, suplementadas com MPDR e MNPDR. O teor de gordura do leite foi de 2,39; 2,12 e 1,89% paras vacas suplementadas com MPDR, MNPDR e controle, respectivamente. A produção diária de gordura foi 0,57; 0,58 e 0,58 kg/dia e a produção diária de leite corrigido para gordura (3,5%), 21,25; 21,19 e 21,35 kg/dia, para os respectivos tratamentos controle, MPDR e MNPDR. A suplementação com MPDR não alterou a produção de leite, porém melhorou a sua composição no início da lactação.

Pitiose cutânea equina de localização atípica tratada topicamente com solução de anfotericina B e DMSO

Background: Cutaneous lesions by Pythium insidiosum infection are commonly observed in horses, especially in those living at flooded environments. Equine pythiosis is characterized by the development of tumoral masses that are frequently located at distal limbs, ventral abdomen, thorax, breast and face. The lesions are usually granulomatous, serosanguineous and ulcerated, most often destroyed by self-mutilation due to the intense pruritus. The proposed treatment includes surgical excision followed by antifungal drugs administration, which can be done systemically or topically. Amphotericin B and dimethyl sulfoxide (DMSO) in association has been successfully used for cutaneous pythiosis topical treatment due to the DMSO property to carry any substance through plasmatic membranes.Case: The present report concerns a 12-year-old mixed breed gelding presenting with self-mutilation of a tumoral mass located at the left flank. The owners reported that the horse had initially presented a small wound that had evolved to a 20-cm in diameter mass in 4 weeks. Tissue samples were collected, processed and stained by the Gomori's methenamine silver (GMS) method. The histopathological analysis revealed Pythium insidiosum hyphae in a granulomatous tissue, especially located at peripheral region, where kunkers were present. Surgical excision of the mass followed by cauterization was indicated as initial treatment, and due to financial reasons, the owners elected only the topical antifungal therapy to control the fungus infection after surgery. Flunixin meglumine was also administrated for five days aiming the control of pain and inflammation. The wound was cleaned with povidone-iodine solution and rinsed with a solution containing, 50 mg, of amphotericin B in 10 mL of sterile water and 10 mL of DMSO. This procedure was carried Out twice a day. The wound healed fast due to an excellent centripetal epithelialization. and the horse was discharged after 64 days showing only 5% of the initial wound area. The owner reported by telephone the complete healing and hair growth 10 days after discharge.Discussion: Despite the atypical location of the tumoral lesion described at the present report, the history and clinical manifestations, especially the intense pruritus, showed similarity with other characteristic reports of equine cutaneous pythiosis. The diagnosis was confirmed by the histopathological examination showing hyphae structures, as described to be evidences of the presence of Pythium insidiosum in the tissue. The surgical procedure was the first step to provide remission of clinical signs, and one day after surgery the pruritus desapeared. After excision of the granulomatous tissue and cauterization, daily topical administration of amphotericin B associated with DMSO was effective in destroying the infectious agent, as observed by the excellent epithelization. A pink granulation tissue grew up providing an ideal surface for epithelial migration and the healing process progressed quickly. Centripetal epithelialization reduced the wound area until 3% of the initial area in 64 days of treatment, when the remaining wound was found almost completely healed and covered with hair. At the present report, the horse presenting pythiosis was only topically treated. The recommended therapy using amphotericin B and DMSO solution was effective, economically viable and low risk, considering that the systemic antifungal therapy usually suggested is expensive and extremely nephrotoxic. The atypical location of the lesion on the left flank shows that any anatomical region can be affected by the fungus, since the conditions for its development were present.

Diferentes critérios de formulação de rações para frangos de corte no período de 1 a 21 dias de idade

Dois experimentos foram realizados com o objetivo de avaliar diferentes critérios de formulação de rações, baseados em aminoácidos totais e digestíveis, para frangos de corte no período de 1 a 21 dias de idade. As dietas foram formuladas para atender às exigências de metionina, metionina+cistina, lisina e treonina. No primeiro experimento, foram utilizados 800 pintos machos, Hubbard, distribuídos em delineamento inteiramente casualizado com 4 critérios de formulação, para atender às exigências em aminoácidos totais - NRC (1994), Rostagno et al. (1992), Degussa (1997) e AEC (1993), com 4 repetições de 50 aves. No segundo experimento, foram utilizados 600 pintos machos, Cobb, distribuídos em delineamento inteiramente casualizado com 3 critérios de formulação, para atender às exigências em aminoácidos digestíveis - Rostagno et al. (2000), Baker & Han (1994) e Degussa (1997), com 4 repetições de 50 aves. Foram avaliados ganho de peso, consumo de ração e conversão alimentar. Os resultados obtidos demonstraram não haver diferença no desempenho das aves ao serem alimentadas com diferentes critérios de formulação baseados em aminoácidos totais. Ao formular dietas com aminoácidos digestíveis, o padrão estabelecido por Baker & Han (1994) resultaram em melhor desempenho das aves no período avaliado.

Avaliação do Desempenho e Excreção de Cálcio em Duas Linhagens de Frangos de Corte, na Fase Inicial, Alimentados com Diferentes Níveis de Aminoácidos e de Cálcio

Dois experimentos foram realizados com o objetivo de avaliar os efeitos de diferentes níveis de aminoácidos e de cálcio sobre o desempenho e excreção de cálcio de duas linhagens de frangos de corte (Cobb e Avian Farms) na fase inicial de criação. Foram avaliados os dados de desempenho (consumo de ração, ganho de peso e conversão alimentar) e excreção de cálcio. Foram utilizadas 540 aves, em cada experimento, distribuídas em delineamento inteiramente casualizado, em esquema fatorial 3x2, ou seja, 3 perfis de aminoácidos (metionina, metionina+cistina, lisina e treonina - 100%, 125% e 150% dos níveis do NRC,1994) e 2 níveis de cálcio (75% e 100% dos níveis do NRC,1994), com 30 aves por repetição. Analisou-se o cálcio excretado das aves, e para tanto, utilizou-se 72 aves, em cada experimento, alojadas em gaiolas, seguindo o mesmo delineamento descrito anteriormente. Não houve interação entre os tratamentos estudados. Para a linhagem Avian Farms, o ganho de peso diminuiu com a elevação dos níveis de aminoácidos e ocorreu menor excreção de cálcio com menor nível de cálcio da dieta. A linhagem Cobb não foi influenciada pelos tratamentos em nenhum dos parâmetros estudados.

Exigências de metionina + cistina digestível para aves de corte ISA Label criadas em semiconfinamento

Foram realizados três experimentos para determinação das exigências de metionina+cistina (met+cis) digestível para aves da linhagem ISA Label. As aves foram criadas em semiconfinamento nas fases inicial (1 a 28 dias), crescimento (28 a 56 dias) e final (56 a 84 dias). em cada experimento, foram utilizadas 480 aves (metade de cada sexo) alojadas em 24 piquetes. O delineamento experimental utilizado foi o inteiramente casualizado, em esquema fatorial 4 × 2 (níveis de met+cis e sexos) com três repetições de 20 aves. Os níveis de met+cis digestível avaliados foram: 0,532; 0,652; 0,772; 0,892% na fase inicial; 0,515; 0,635; 0,755; 0,875% na fase de crescimento; e 0,469; 0,589; 0,709; 0,829% na fase final. Foram avaliados o desempenho, as características de carcaça, a deposição de proteína e gordura corporal, o peso e o teor de proteína das penas. Na fase inicial, os níveis de met+cis digestível na ração recomendados para machos e fêmeas foram 0,76 e 0,80%, que correspondem a 0,252 e 0,268% de met+cis por Mcal de energia metabolizável da ração, respectivamente. Para aves ISA Label na fase de crescimento, recomenda-se 0,716% de met+cis digestível na ração, independentemente do sexo, que corresponde a 0,235% de met+cis por Mcal de em da ração. Na fase final, recomendam-se níveis de met+cis digestível de 0,756 e 0,597%, que correspondem a 0,244 e 0,193% de met+cis por Mcal de energia metabolizável na ração para machos e fêmeas, respectivamente.

Desempenho, rendimento de carcaça e excreção de cálcio de frangos de corte alimentados com diferentes níveis de aminoácidos e cálcio no período de 22 a 42 dias de idade

Um experimento foi conduzido com o objetivo de avaliar diferentes níveis de aminoácidos e de cálcio para frangos de corte no período de 22 a 42 dias de idade. Avaliou-se o desempenho, o rendimento de carcaça e a excreção de cálcio. Foram utilizadas 540 aves, machos, da marca comercial Cobb, distribuídas em delineamento inteiramente casualizado, em esquema fatorial 3x2, ou seja, 3 níveis de aminoácidos (metionina, metionina+cistina, lisina e treonina - 100%, 125% e 150% NRC,1994) e 2 níveis de cálcio (75% e 100% NRC,1994), com 3 repetições de 30 aves cada. Os aminoácidos estudados foram metionina, metionina+cistina, lisina e treonina. O desempenho das aves foi avaliado pelo peso vivo, ganho de peso, consumo de ração e conversão alimentar. O rendimento de carcaça e a percentagem de cada parte foram avaliados de 2 aves de cada repetição, escolhidas ao acaso, perfazendo um total de 6 animais por tratamento. O rendimento de carcaça foi realizado após o abate levando-se em consideração a percentagem de peito, pernas (coxa+sobrecoxa), asas, dorso, cabeça+pescoço e pés em relação à carcaça eviscerada. Além disso, analisou-se o cálcio excretado pelas aves. Para tanto, utilizou-se 72 aves, alojadas em gaiolas, seguindo o mesmo delineamento descrito anteriormente. Não houve interação entre os fatores estudados para as características avaliadas. As que consumiram dietas com 125% e 150% de aminoácidos apresentaram melhor conversão alimentar. O rendimento de peito foi afetado pelos níveis de cálcio.

The contribution of genes required for anaerobic respiration to the virulence of Salmonella enterica serovar Gallinarum for chickens

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