Mini-dissertation was submitted in partial compliance with the requirements for the Master’s Degree in Technology: Homoeopathy, Durban University of Technology, 2012. / It was reported by the International Diabetes Federation (IDF) Diabetes Atlas, in 2003, that a prevalence figure of 3.4% exists for the 24 million South Africans between the ages of 20 and 79, with an expected increase to 3.9% by 2025. Considering that patients with diabetes are at increased risk of cardiovascular disease, blindness, amputation and renal failure it is therefore not surprising that the costs associated with diabetes are estimated to increase worldwide. It is clear that according to the current trends in dietary and exercise practices, South Africa will be affected by the rise in obesity and subsequent diabetes mellitus. It is critical that a concerted effort involving all parties concerned be made to combat this rapidly increasing problem (Rheeder, 2006:20). AIM The purpose of this double-blind, randomized clinical trial was to compare the effectiveness of Momordica charantia homoeopathic mother tincture as compared to Momordica charantia 6CH, in the treatment of type 2 diabetes mellitus in patients on Metformin. METHODOLOGY Thirty patients were recruited and were selected for the study on the basis of inclusion and exclusion criteria. These participants were then randomly and equally divided into two groups. Each participant attended a total of four consultations with the researcher, over a two month period, at the Durban University of Technology (DUT) Homoeopathic Day Clinic. At the commencement of the first consultation, each participant received the subject information letter (Appendix A) for perusal and the informed consent form (Appendix B) to sign. Following this, the researcher took a full, detailed iv case history (Appendix F) and performed a physical examination (Appendix G) of each patient. Participants were required to have a Glycosylated haemoglobin (HbA1C) test performed following the first and fourth consultations. Participants were also required to complete daily Log Sheets (comprising self administered fasting blood glucose readings using issued Bayer Ascensia Elite Glucometers) for the entire duration of the study (8 weeks). SPSS version 18 was used to analyse the data. A p value < 0.05 was considered as statistically significant. The time effect was assessed for intra-group comparison whereas the time x group treatment effect was assessed for inter-group comparison. Means were calculated for both fasting blood glucose and glycosylated haemoglobin for the two respective groups and tabulated in order to describe the data obtained (Descriptive statistics). RESULTS Both groups reflected a statistically non significant decrease in fasting blood glucose levels with no significant differences between the two groups when comparing reduction in fasting blood glucose levels. Group 1 (Momordica charantia homoeopathic mother tincture) reflected a non significant increase in glycosylated haemoglobin (HbA1C) levels while Group 2 (Momordica charantia 6CH) reflected a statistically significant increase over time in HbA1C levels. There were no significant differences between the two groups when comparing reduction in HbA1C levels. / M

Caenorhabditis elegans has been studied as a model organism in various areas of biomedical research because it shares many conserved functions at molecular and genetic levels with humans. Specifically, it is an ideal organism to study heterogeneous metabolic syndromes such as Type 2 Diabetes Mellitus (T2DM) as C. elegans can be used to delineate molecular pathways that are at the core of its problems. A growing number of populations worldwide are faced with chronic T2DM, which also manifests several complications, such as blindness, neuropathy and cardiovascular diseases. Currently, metformin is the first-line drug of choice administered to treat T2DM. While the mechanism by which it alleviates the symptoms of diabetes is unknown, it has been found to reduce metabolic rate by partially inhibiting the mitochondrial complex I in mammals. Using C. elegans as a genetic model organism, we show that metformin reduces the mitochondrial activity through endosomal Na+/H+ exchanger, which a previous lab member has found to be a potential target of metformin. Furthermore, we show that high glucose diet−known to reduce the worm’s lifespan−alter the endosomal-lysosomal system and autophagy, providing insights to using C. elegans as a diabetic model. Based on these results, we propose that C. elegans can serve as a model organism to study T2DM as well as provide new ways to further investigate the pathophysiology of this disease.

The extract from the plant Galega officinalis containing the guanidine derivative galegin has been used in the treatment of diabetes-associated complications since middle ages. Nevertheless, the positive effects of the treatment were often overweight by the adverse side effects. Some sixty years ago guanidin was replaced by the less toxic synthetic biguanide derivatives - metformin, phenphormin and buformin, the latter two being withdrawn due to the unacceptable risk of fatal lactate acidosis. Metformin is still widely used antidiabetics and belongs to the first choice drugs in the treatment of type 2 diabetes. Phenphormin is now gaining renewed attention with regard to its antineoplastic properties. Despite its long-term clinical use the mechanism of biguanides action is not fully understood yet. At present it is generally accepted that the core of its antihyperglycemic effect lays in the inhibition of hepatic gluconeogenesis. In contrast, there is less consensus regarding the particular metabolic pathway or target that are responsible for the metformin-induced attenuation of gluconeogenesis. For a long time, a hot candidate for metformin target in the cell was AMP-activated kinase (AMPK) but the metformin effect was proved also in mice carrying the dominant negative mutation of AMPK α subunit. Quite...

A influência da obesidade e da resistência à insulina (induzidas em ratos por injeção de glutamato monossódico em neonatos) sobre o desenvolvimento tumoral (5x105 células do tumor de Walker-256) e os efeitos da metformina (300mg/kg, v.o., 15 dias) nessa condição foram investigados. Na 16ª semana de vida, inocularam-se as células e iniciou-se o tratamento. Após 15 dias de tratamento, caracterizou-se a obesidade e a analisou-se o crescimento tumoral. O desenvolvimento tumoral e a caquexia foram maiores nos obesos. A metformina reduziu o desenvolvimento do tumor, mas não a caquexia. Apesar da metformina não ter melhorado a sensibilidade à insulina, corrigiu a dislipidemia, reduziu a peroxidação lipídica e as gorduras periepididimal e retroperitoneal. Conclui-se que a obesidade aumenta o desenvolvimento tumoral e que a metformina é eficaz em diminui-lo. O mecanismo envolvido parece não depender da melhora da sensibilidade à insulina / The influence of obesity and insulin resistance (induced in rats by monosodium glutamate in neonates) on tumor development (5x105 Walker-256 tumor cells) and the effect of metformin (300mg/kg, by gavage, for 15 d) on it. On the 16th week, tumor cells were subcutaneously injected and the treatment started. On the 18th week, the obesity was characterized and the tumor was evaluated. The tumor development and the cachexia were higher in obese rats. The tumor development was reduced by metformin, but not cachexia. Although metformin did not improve insulin sensitivity it did correct the dislypidemia, reduced the periepididimal and retroperitoneal adipose tissues and lipid peroxidation. In conclusion obesity increases tumor development and metformin is able to reduce it. The reduction occurred independently of the correction of insulin resistance

Avaliou-se a participação do óxido nítrico (NO), do fator hiperpolarizante derivado do endotélio (EDHF), dos produtos da ciclooxigenase (COX), das espécies reativas de oxigênio (EROs) e o efeito do tratamento com metformina (Met) nas alterações vasculares em ratos com obesidade induzida por glutamato monossódico (MSG). O tratamento com Met corrigiu alterações metabólicas em ratos MSG, reduzindo o acúmulo de gordura visceral, corrigindo a resistência à insulina, a hiperinsulinemia e a dislipidemia. Ratos MSG apresentaram aumentada resposta contrátil e diminuída sensibilidade à Ach, associadas a alterações na via do NO, do EDHF, dos produtos da COX e das EROs. Ratos MSG com 16 semamas apresentaram hiperresponsividade ao nitroprussiato de sódio, que foi mantida nos ratos tratados com Met. A Met corrige as alterações da resposta vascular atuando sobre o NO e o EDHF, reduzindo a geração de EROs e interferindo na resposta do músculo liso vascular, mantendo a hiperresponsividade ao NO. / The role of the nitric oxide (NO), the endothelium derived hyperpolarizing factor (EDHF), the ciclooxygenase (COX) products and the reactive oxygen species (ROS), as well as the effect of metformin (Met) treatment on the vascular alterations in rat model of obesity induced by monosodium glutamate (MSG) were evaluated. Met treatment corrected metabolic alterations in MSG, reducing fat accumulation, correcting dyslipidemia, insulin resistance and hyperinsulinemia. MSG rats had an increased response to norepinephrine and decreased sensitivity to acetilcholine, which were associated with alterations in NO, COX products and ROS. Sixteen-week-old MSG rats presented hyperresponsiveness to sodium nitroprusside, which was preserved in Met-treated group. Met corrects the alterations of the vascular reactivity acting on NO and EDHF, and decreasing the ROS generation, besides its effect on the vascular smooth muscle response, preserving the hyperresponsiveness to NO.

A influência da obesidade induzida por glutamato monossódico em ratos sobre o desenvolvimento do tumor de Walker-256 e os efeitos da metformina (300mg/kg, v.o., 15 dias) foram investigados. Na 16ª semana de vida, inocularam-se as células tumorais e iniciou-se o tratamento. Após 15 dias, analisou-se o crescimento tumoral. A viabilidade de células tumorais, MCF-7, tratadas com meformina foi avaliada. A obesidade contribuiu para maior desenvolvimento tumoral e reduziu a sobrevida dos ratos. A metformina foi eficaz em impedir o aumento do tumor e aumentou a sobrevida dos ratos. Teve efeito antiproliferativo sobre as células MCF-7, efeito esse relacionado ao bloqueio do ciclo celular, estresse oxidativo, aumento na apoptose e necrose celulares, e aumento na atividade da AMPK e do FOXO3a. Conclui-se que a obesidade contribui de maneira significativa para o desenvolvimento tumoral e que a metformina é eficaz em diminuí-lo. / The influence of obesity induced by monossodium glutamate in rats on Walker-256 tumor development and the metformin (300/kg, b.w., 15 days) effect were analyzed. At the 16th week, 1x107 Walker-256 tumor cells were inoculated and the treatment with metformin was started. Following this treatment, the tumor growth was analyzed. The cell viability of MCF-7 cells treated with metformin was analyzed. Obesity positively contributed for tumor development and it reduced life span of the rats. Metformin reduced the tumor development and increased the life span of the rats. It presented an antiproliferative effect in MCF-7 cells. Effect associated with an increase in oxidative stress, apoptosis, necrosis and cell cycle arrest in G0-G1 phase. Furthermore, its effect is associated with an increased in AMPK and FOXO3a activities. In conclusion, obesity contributed significantly to tumor development and metformin is effective in reduced it.

A diabetes mellitus (DM) é um grupo de doenças metabólicas caracterizadas por hiperglicemia resultante do déficit na secreção e/ou ações de insulina. Dentre as muitas complicações da diabetes incluem a osteopenia diabética, que causa osteoporose e aumento do risco de fraturas ósseas. A patofisiologia da baixa resistência óssea associada a DM é considerada multifatorial, podendo ser decorrente da deficiência de insulina, resistência à insulina, insuficiência de osteoblastos, deficiência de vitamina D, formação e acúmulo dos produtos finais da glicação avançada e complicações microvasculares. Por isso, existe um interesse crescente no estudo da diabetes associada a outras alterações metabólicas e o efeito das drogas antidiabéticas, de forma a reverter os efeitos maléficos. O objetivo desse estudo foi avaliar a influência das drogas antidiabéticas na movimentação dentária ortodôntica e na densidade/microarquitetura óssea alveolar em ratos diabéticos. Assim, ratos Normoglicêmicos (NG,n=20) e Diabéticos induzidos pela estreptozotocina (DM1,n=60) foram divididos em: TinDM1(n=20) tratados com Insulina, TinmetDM1(n=20) tratados com Insulina+Metformina, e os STDM1(n=20) e STNG(n=20) que não receberam tratamento. Após 14 dias da indução, o 1o molar superior direito recebeu força ortodôntica (50g) em sentido mesial. Nos periodos experimentais de 0, 3, 7 e 14 dias, as maxilas foram coletadas e submetidas às análises microtomográficas para quantificar a movimentação dentária e a densidade óssea e histológica, para avaliar as alterações periodontais ocorridas durante a movimentação. Os dados microtomográficos foram submetidos à ANOVA a dois critérios e teste de Tukey (p<0,05). A indução com estreptozotocina induziu ao quadro de diabetes grave (glicemia de jejum de 325mg/dL) os quais foram acentuados com o tempo no grupo STDM1 (404mg/dL). A utilização de insulina e da associação insulina e metformina reduziram consideravelmente os níveis glicêmicos (127mg/dL). A força de 50g aplicadas no 1o molar promoveu movimentação dentária linear, sendo menor no grupo STNG (116&#x3BC;m) e maior nos diabéticos (173&#x3BC;m). Durante a movimentação a densidade óssea no grupo STNG foi mantida (BV/TV=83%), enquanto nos TinDM1 e TinmetDM1 ocorreu pequena redução (BV/TV=76%). Já nos STDM1 a força produziu grande perda óssea (BV/TV=62%). No grupo STDM1 O quadro histopatológico confirma os efeitos deletérios nas estruturas dentarias e periodontais durante a movimentação dentária, com acentuada perda óssea e processo inflamatório. A redução dos índices glicêmicos com utilização das drogas hipoglicemiantes nos grupos TinDM1 e TinmetDM1 equilibrou o processo de formação e reabsorção óssea no STNG, após os 3 dias da movimentação. Concluímos que, a utilização contínua de insulina ou insulina e metformina nos animais diabéticos diminuem significativamente o quadro de perda óssea alveolar decorrente das forças ortodônticas no estado diabético. / Diabetes mellitus (DM) is a group of metabolic diseases characterized by hyperglycemia resulting in deficits in the secretion and/or insulin action. Among the many complications of diabetes, it includes diabetic osteopenia that causes osteoporosis and increased risk of bone fractures. The pathophysiology associated with low bone strength in DM is considered multifactorial and may be due to insulin deficiency, insulin resistance, osteoblast deficiency, vitamin D deficiency, formation and accumulation of advanced glycation end products and microvascular complications. Therefore, there is a growing interest in the study of diabetes associated with other metabolic abnormalities and the effect of antidiabetic drugs, in order to reverse the deleterious effects. The objective of this study was to evaluate the influence of antidiabetic drugs in orthodontic tooth movement and alveolar bone density/microarchitecture in diabetic rats. Thus, normoglycemic rats (NG, n=20) and streptozotocin-induced diabetic (DM1, n=60) were divided into TinDM1 (n=20) treated by insulin, TinmetDM1 (n=20) treated by Insulin + Metformin and STDM1 (n = 20) and STNG (n = 20) that received no treatment. After 14 days of induction, the M1 received orthodontic force (50g) to move mesially. After 0, 3, 7 and 14 days jaws were collected and subjected to microtomographic images analysis to quantify, tooth movement and bone density and histological analysis to evaluate periodontal changes occurred during the movement. Microtomographic data were submitted to two-way ANOVA and Tukey test (p <0.05). The induction with streptozotocin induced severe diabetes frame (fasting blood glucose 325 mg/dL) which accentuated over the time of the disease in STDM1 group (404mg/dL). The use of insulin and insulin and metformin reduced blood glucose levels to satisfactory values (127mg/dL). The strength of 50g applied on M1 promoted linear tooth movement, being lower in STNG group (116&#x3BC;m) and higher in diabetics (173&#x3BC;m). When handling bone density was maintained at STNG group (BV/TV = 83%), in TinDM1 and TinmetDM1 small reduction occurred (BV/TV = 76%). Already in STDM1 the force produced large bone loss (BV/TV = 62%). Histopathological analysis confirmed the deleterious effects on periodontal and dental structures during tooth movement in STDM1 group, with marked bone loss and inflammatory process. Reducing the glycemic index by using insulin in TinDM1 group and insulin + metformin in TinmetDM1 balanced process of bone formation and resorption after 3 days of mechanical loading. We conclude that the continuous use of insulin or insulin and metformin in diabetic animals diminishesalveolar bone loss resulting from orthodontic forces in the diabetic condition.

Background Glioblastoma Multiforme (GBM) is the most commonly occurring tumour of the central nervous system (CNS). Currently GBM is considered an incurable malignancy with patients experiencing abysmal life expectancies. Lack of progress in the discovery of novel treatments has led to the repurposing of existing licenced medication as a possible alternative option. Metformin is from the biguanide family of drugs and is the most common medication used in the treatment of type 2 diabetes. Clinical studies have reported that, in type 2 diabetic patients, metformin might reduce cancer incidence and severity. Currently, metformin is being assessed in clinical trials as a treatment for a range of cancer types including GBM. The antineoplastic mechanisms utilized by metformin and other biguanides have not been fully elucidated. Methods The effects of metformin were evaluated, alone and in combination with other agents, on a panel of GBM cell cultures. Functional analysis of metformin mechanism of action was assessed through measurement of apoptosis, depolarisation of the mitochondria membrane, caspase pathway activation, cell cycle progression and the expression levels of micoRNAs. Results Analysis of fourteen GBM cell cultures showed a cytotoxic response to metformin that was significantly linked to the P53 status (p=0.0024). In combination drug testing, one of the four drugs showed a synergistic pairing with metformin. The kinase inhibitor sorafenib, showed synergism (CI ≤ 1) in eight GBM cell cultures. Flow cytometry of metformin treated GBM cells showed no significant increase (p > 0.005) in apoptotic cell populations. Caspase 3/7 levels showed no significant increase post metformin treatment (p > 0.005). Metformin caused depolarisation of the mitochondrial membrane in six GBM cell cultures. Four microRNAs were shown to have expression levels changes post-metformin treatment. Upregulation of expression was identified in miR-140, miR-192, let-7c. Downregulation was identified in miR-222. Conclusions Metformin was shown to have cytotoxic effect on a GBM cell cultures and has potential as GBM therapeutic agent and possible treatment synergy with sorafenib. The significance of P53 status to metformin sensitivity may suggest that its use should be directed to a sub-set of GBM patients. Mechanism for cell death by metformin was shown not to rely on apoptotic pathways but caspase 3/7 independent depolarization of mitochondrial cell membranes and cell cycle arrest. Investigations into autophagy may help to further define the pathways metformin is utilising to promote cell death. The impact of metformin on the expression profile of miR-222, miR-192 and let-7c is in line with clinical studies of other cancer types. This shows possible insight into the cancer independent actions of metformin. The interplay recorded between glucose availability and cell death indicates a possible key factor in the utilisation of metformin as a therapeutic agent. This finding may warrant the addition of dietary control regimes in clinical trials to maximise metformin efficacy. This work highlights the strong potential for biguanides in the development of new drug treatments and in expanding our knowledge of cancer metabolism.

INTRODUÇÃO: Estudos observacionais e pré-clinicos sugerem o efeito antitumoral da metformina em tumores sólidos, incluindo o câncer colorretal. Entretanto, os efeitos da metformina no câncer colorretal ainda não foram testados em estudos prospectivos. PACIENTES E MÉTODOS: Este foi um estudo de fase II, unicêntrico, braço único de pacientes com câncer colorretal metastático em progressão e previamente tratados com 5-FU, irinotecano, oxaliplatina e anti-EGFR, se RAS selvagem. Os pacientes receberam metformina 850 mg VO duas vezes por dia e 5-FU 425 mg/m2 e leucovorin 50 mg IV semanal até progressão de doença, toxicidade inaceitável ou retirada de consentimento. O desfecho primário foi controle de doença em 8 semanas. RESULTADOS: Dos 50 pacientes incluídos, 11 (22%) alcançaram o desfecho primário. Para toda coorte, a SLP foi de 1,8 meses e a SG mediana de 7,9 meses. Quando avaliamos somente os 11 pacientes que alcançaram controle de doença na semana 8, a SLP foi de 5,6 meses e a SG foi de 16,2 meses. Houve tendência a maior sobrevida entre os obesos (12,4 vs 5,8 meses de acordo com IMC maior ou menor que 30) e aqueles que tiveram maior intervalo livre de 5FU antes de entrar no estudo. O tratamento foi bem tolerado e os principais efeitos colaterais de qualquer grau foram diarreia, náusea, vômito e mielotoxicidade. CONCLUSÃO: Neste estudo de fase II, a metformina combinada ao 5FU apresentou atividade modesta na população geral de pacientes com câncer colorretal metastático refratário. Em análise de subgrupo, obesos e maior intervalo livre de 5FU foram associados a controle de doença prolongado. Estudos prospectivos randomizados com metformina em câncer colorretal devem ser realizados / BACKGROUND: Observational and pre-clinical studies have suggested that metformin has antitumor effects in solid tumors, including colorectal cancer. However the effects of metformin in colorectal cancer have not been tested in clinical trials. PATIENTS AND METHODS: This was a single center, single-arm phase II clinical trial where histologically confirmed colorectal cancer patients with measurable and progressing metastatic disease previously treated with 5-FU, irinotecan, oxaliplatin and an anti-EGFR, if the tumor was RAS wild type, were enrolled to receive metformin 850 mg orally bid continuously plus 5-FU 425 mg/m2 and leucovorin 50 mg IV weekly until disease progression, unacceptable toxicity or consent withdrawn. The primary endpoint was disease control rate at 8 weeks. RESULTS: Among 50 patients included, 11 (22%) met the primary endpoint. The median progression free survival was 1.8 months and the median overall survival was 7.9 months. Analyzing only those 11 patients who achieved disease control rate at 8 weeks, their median progression-free survival was 5.6 months and their median overall survival was 16.2 months. There was a trend for prolonged median survival for obese patients (12.4 vs 5.8 months) and those longer off 5FU. The treatment was well tolerated and the main side effects were diarrhea, nausea, vomiting and myelotoxicity. CONCLUSION: Metformin and 5FU showed an overall modest but intriguing activity in refractory colorectal cancer patients in this phase II study. Some patients presented long-term disease control. Further trials are needed to confirm these results, particularly in obese patients with colorectal cancer

O câncer é um problema de saúde pública mundial e pode causar anorexia. Mediadores inflamatórios desempenham importante papel no desenvolvimento da anorexia relacionada à doença. Estudos realizados com carnosina (&#946;-alanina-L-histidina) e metformina demonstraram eficiente ação anti-inflamatória e antitumoral. O objetivo deste estudo foi analisar a ação da metformina e &#946;-alanina no controle do crescimento tumoral e produção das citocinas, bem como a secreção dos neuropeptídios. Foram utilizados 25 camundongos machos, divididos em grupo controle (C), controle tumor (CT), tumor suplementado com &#946;-alanina (TB), tumor administrado com metformina (TM) e tumor administrado com metformina+&#946;-alanina (TMB). TM demonstrou redução no peso e ingestão alimentar comparado aos C e TMB. TB apresentou maior massa tumoral que os demais grupos. Ainda, o CT apresentou maior expressão da inflamação em relação aos demais grupos. Concluímos que os tratamentos utilizados demonstraram um potencial efeito benéfico frente ao tumor, sendo uma estratégia coadjuvante para o tratamento do câncer. / Cancer is a worldwide public health problem and can cause anorexia. Inflammatory mediators play an important role in the development of disease-related anorexia. Studies with carnosine (&#946;-alanine-L-histidine) and metformin have demonstrated effective anti-inflammatory and antitumor action. The objective of this study was to analyze the action of metformin and &#946;-alanine without control of tumor growth and cytokine production, as well as a neuropeptide secretion. Twenty-five male mice, tumor control (CT), tumor-supplemented with &#946;-alanine (TB), tumor administered with metformin (TM) and tumor with metformin + &#946;-alanine (TMB) were used. TM demonstrated a reduction in weight and food intake compared to C and TMB. TB presented greater tumor mass than the other groups. Still, CT showed a greater expression of inflammation in relation to the other groups. We conclude that the treatments used demonstrated a potential beneficial effect against the tumor, being a supporting strategy for the treatment of cancer.