Hradecká M.: Natural compounds applicable for the treatment of metabolic syndrome, Diploma thesis 2018/2019, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany, pp. 60. Occurence of metabolic syndrome is increasing worldwide in children and adults. The use of natural compounds is one of possibilities in the prevention and treatment of metabolic syndrome. These compounds may act complexly or affect individual risk factors associated with metabolic syndrome. Selected plants with these effects are incorporated into my diploma thesis (for example Allium sativum, Crataegus laevigata, Hibiscus sabdariffa, Persea americana, Rosmarinus officinalis, Silybum marianum and Vaccinium myrtillus). The alga Undaria pinnatifida and fungus Pleurotus sajor-caju are also mentioned. It is necessary to carry out other clinical studies, where the positive effect of mentioned constituents will be confirmed, which could be added to the list of phytopharmaceuticals used to prevent, slow down or treat the metabolic syndrome in the future. Keywords: natural compounds, metabolic syndrome

Second-generation antipsychotics (SGAs) are prescribed for treatment of psychosis. A major side effect of SGAs is an increased risk of metabolic syndrome (MetS) with symptoms of hypertension, hyperlipidemia, hyperglycemia, and truncal obesity. A clinic in the northeastern United States was not screening patients for MetS when being treated with SGAs. The purpose of this project was to educate staff on MetS risk factors, signs, symptoms, and patient management with a goal to improve their knowledge of MetS. Lewin's change theory provided a conceptual framework for the project. The project question explored the development and evaluation of an educational module on MetS increased staff knowledge. Educational content was guided by current literature and the American Psychiatric Association and American Diabetic Association practice guidelines. Five expert panel members, consisting of 3 psychiatrists, an advance practice nurse, and a registered nurse reviewed the education program and evaluated content using a Likert-type questionnaire. Expert panel evaluations indicated that the module content contained useful clinical information on MetS screening for patients on SGAs. After panel review, the program was presented to 7 clinic staff. Pretest and posttest questionnaires asked 10 multiple choice questions and results were compared. Questions on SGA side effects, MetS complications, prevalence, baseline assessment measures, lab work, and needed collaboration were answered correctly by 6 of the participants pretest and all questions after receiving the education program. The project has the potential to promote positive social change through staff education on MetS screening for patients, thus improving patient outcomes.

Metabolic Syndrome (MetS) is a collective term for a cluster of disorders, including dysglycemia, central obesity, dyslipidemia, hypertension, and eventual end organ damage. The combination of these disorders increases the risk of many kinds of end organ damages, including coronary heart disease, kidney failure, and cirrhosis. MetS is highly prevalent in the United States, affecting one third of the U.S. population in a 2009 estimate. The Lyon strains are three rat strains selectively inbred from the same colony of outbred rats for different blood pressure levels. The Lyon Hypertensive (LH) strain, in addition to its essential hypertension phenotype, also harbors many disorders found in MetS. The Lyon Normotensive (LN) rat strain is completely devoid of these symptoms, while Lyon Low-pressure (LL) is obese but is resistant to other traits of MetS. Rat chromosome 17 (RNO17) has previously been linked with many of MetS' phenotypes in Lyon Hypertensive (LH). In this project, we are using a mixture of genetical genomics and systems biology methods to identify genetic elements that may cause the LH phenotype. Divergent haplotype blocks between the Lyon strains were first identified by the analysis of the distribution of observed strain differences (OSD) calculated from the result of genome resequencing. Divergent haplotype regions totaling less than 16% of the rat genome that contain more than 95% of the identified SNPs in each of the three pairwise comparisons between the Lyon strains have been identified; in particular, there are 14 divergent haplotype blocks between LH and LN spanning 7.7% of RNO17 that harbor more than 97% of SNPs identified on RNO17. Twenty-five genes in these regions were thus identified as potential genetic determinants for MetS. Phenotypic QTLs (pQTL) and expression QTLs (eQTL) mapping from a cohort of male LH × LN F2 rats were performed by putting the cohort on a 15-week phenotyping protocol and genome-wide genotyping. Total liver RNA from 36 individuals from the cohort were sequenced to provide expression data for eQTL mapping. We have mapped 22 pQTLs that are statistically linked to 15 traits, with RNO17 linked to 15 traits associated with blood pressure, leptin and body weight. We have also identified 1,200 eQTLs from this cohort, including 11 eQTLs with cis-linkage with one or more genes. On RNO17, we have identified two SNPs between 29-39 Mb which are significantly linked to the expression of 85 genes; the only gene with cis-linkage with these SNPs, RGD1562963, was hence identified as a putative master regulator. Transcriptome analyses were then performed on the Lyon parental animals; the total liver and kidney of RNA from 6 each of LH, LL and LN strains that were subjected to the same 15-week phenotyping protocol were sequenced for differential expression analysis, gene coexpression network analysis and quantitative trait transcript analysis. Differential expression analysis identified 4 genes on RNO17's divergent haplotype regions: Cul2 and the aforementioned RGD1562963 for liver, Amph and Bambi for kidney. Quantitative trait transcript analyses have shown significant correlations between the expressions of these four genes with one or more of the traits of the animals treated, validating their status as potential genetic determinants for MetS. However, out of the 84 genes that RGD1562963 potentially regulates, only two other genes (Cul2 and Supt4h1) have significant correlations with one or more traits. Gene coexpression network analyses have shown a relationship between genes on the TGF-β pathway and the differentially expressed genes in the kidney, supporting our speculation on the hyperactivity of the TGF-β system in the etiology of the LH phenotypes. An LH-17LN consomic strain was also generated by introgressing an LN copy of RNO17 onto the LH genomic background to validate in vivo the role of RNO17 in the etiology of MetS symptoms in LH. We have observed that the consomic strain has significantly decreased body weight, adiposity, blood pressure, and inter-week blood pressure differences that may be a surrogate for salt sensitivity. Thus, the role of RNO17 on the LH genotype is validated. In summary, we have been able to identify, by in vivo and in silico methods, that RNO17 is related to the MetS traits in LH; that 4 genes, Amph, Bambi, Cul2, and RGD1562963, are potential genetic contributors to RNO17's effects; and that their effects may include, but are not limited to, the activation of TGF-Β signals.

Thesis (M.Sc. (Biochemistry))--North-West University, Potchefstroom Campus, 2007.

Obesity is today a problem that has reached epidemic proportions. One of the causes of obesity is the over-consumption of energy. Fat is the most energy-dense nutrient, where the quality seems to be more important for the development of the metabolic diseases than the quantity. The fatty acid composition in serum lipid fractions can be used to mirror the dietary fat quality. Stearoyl-CoA-desaturase (SCD) is an enzyme that converts saturated to monounsaturated fatty acids. A surrogate measure of SCD activity can be estimated as a fatty acid ratio; 16:1/16:0 (palmitoleic acid/palmitic acid) and 18:1/18:0 (oleic acid/stearic acid). The aim of this project was to investigate the intra-day variation in the SCD-ratio in humans eating a standardized diet. The results showed that triacylglycerol and nonesterified fatty acid fractions in serum lipids had a significant variance in the 16:1/16:0 ratio during the day, whereas 18:1/18:0 ratio in the same fractions did not exhibit the same pattern. In this study 16:1/16:0 ratio also seems to be a better marker than 18:1/18:0 ratio for estimating SCD activity. For further evaluation of the intra-day variation there need to be a more long-term study of the SCD-activity for a larger group of subjects.

Mexican-Americans are disproportionately burdened by metabolic syndrome, a medical condition characterized by the concurrence of clinical abnormalities that contributes to diabetes, obesity, and cardiovascular disease (CVD). This is alarming since Mexican-Americans constitute two-thirds of the US Latino population, the largest minority and fastest growing group in the US. Investigating acculturative stressors associated with immigration is crucial for eliminating health disparities, but few studies have examined the acculturative impact of Mexican migration to the United States or the relationship between acculturation and metabolic syndrome among Mexican-Americans. The purpose of this dissertation research was to investigate the associations between acculturation and metabolic syndrome among a bi-national sample of Mexicans and Mexican-Americans. Metabolic syndrome was assessed among a bi-national sample of individuals with diabetes using the definition outlined by the International Diabetes Federation, and acculturation was assessed by proxy measures (years lived in the US and generational status) and responses on the Acculturation Rating Scale for Mexican-Americans, version-II. Chi-square, analysis of variance, and logistic regression were used to determine relationships between country, gender, and acculturation status and metabolic syndrome and its biomarkers. The overall prevalence of metabolic syndrome was 79.7%, with 85.0% prevalence among Mexican-Americans and 75.7% among Mexicans (p=0.069). Mexican-Americans had higher blood pressure and central obesity, while Mexicans had higher triglycerides levels. The majority (81.2%) of Mexican-Americans was first generation and lived in the US for an average of 27.65 +/- 16.05 years. The mean acculturation score was -1.83 +/- 1.56, which indicated participants in this study were Mexican-oriented, or more closely associated to Mexican cultural influences than Anglo cultural influences. Higher acculturation scores were positively associated with fasting blood glucose and systolic blood pressure and lower acculturation was negatively associated with fasting blood glucose. Logistic regression analysis showed first generation Mexicans-Americans were more likely to develop metabolic syndrome than second generation Mexican-Americans (OR 7.399, 95% CI 1.464-37.401, p=0.015). Mexican and Mexican-American individuals with type 2 diabetes have a high prevalence of metabolic syndrome, which increases their risk for heart disease and other cardiovascular complications. Mexican-Americans are especially affected by central obesity and hypertension and Mexican immigrants appear to be impacted by negative lifestyle factors upon entering the United States. Acculturation is a complex process and the unclear relationship between acculturation and metabolic syndrome warrants further investigations.

The leading cause of death among women is coronary heart disease (CHD), a multifactorial disease with polygenic heritability estimated at 50%. Polymorphisms in the family with sequence similarity 5, member C’ ( FAM5C) gene have been associated with myocardial infarction (MI), and one single-nucleotide polymorphism (SNP) has partially accounted for linkage in an acute coronary syndrome subset. The linkage peak on FAM5C corresponds directly with a quantitative trait locus for the inflammatory biomarker monocyte chemoattractant protein 1, as well as a linkage peak to metabolic syndrome (MetS). Metabolic syndrome increases the risk of developing CHD, and MI has been positively associated with elevated inflammatory biomarkers. This study was designed as a descriptive pilot gene association study. The purpose was to investigate the variability of the FAM5C SNP (rs10920501) in a cohort of women with documented CHD. It also examined the association between the variability of the FAM5C SNP (rs10920501), MetS, inflammatory markers, and the association with early onset CHD in the presence or absence of MI. A subset of 91 women was derived from an earlier study of women randomized to either a gender-tailored or traditional cardiac rehabilitation program. The results indicated the T allele of FAM5C SNP rs10920501 has a strong protective effect in women with a history of MI. Women with a history of MI and the heterozygous (AT) genotype had a mean age of onset of CHD at 62 years, compared to the homozygous wild type (AA) with a mean age of onset at 55 years, (F (3, 34) = 5.00, p < .01). No women in this study with the homozygous variant (TT) had an MI, further demonstrating the protective effective of the T allele. The genotype of FAM5C SNP rs10920501 explains approximately seven percent of the variability of age of onset of CHD in women who have had an MI, while holding body mass index (BMI) and smoking history constant. There was no significant relationship between FAM5C SNP (rs109320501) and MetS or any inflammatory biomarkers in this sample. In conclusion, FAM5C remains a gene of interest in a complex disease process.

abstract / Pharmacology and Pharmacy / Doctoral / Doctor of Philosophy

Recent studies support the association between metabolic syndrome (MetS), a cluster of cardiovascular risk factors, and diastolic dysfunction. Disproportionate collagen accumulation, particularly cross-linking of collagen, plays a key role in translating interstitial fibrosis into mechanical chamber stiffness and diastolic dysfunction. Characteristic changes in the expression and activity of myocardial lysyl oxidase (LOX), a matrix modifying enzyme that catalyzes cross-linked collagen, are unclear in MetS. We established a diet-induced MetS model to study diastolic dysfunction by treating male C57BL/6 mice a high-fat high-simple carbohydrate (HFHSC) diet for 6 months. Despite blunted gene expression of LOX isoforms, MetS mice demonstrated significant increase in the ratio of protein expression of mature to proenzyme LOX, enhanced LOX activity, and increased cardiac cross-linked collagen compared with controls. This fibrotic response coincided with marked increase in left ventricular end-diastolic pressure and stiffness and impaired diastolic filling pattern. Our data demonstrate that diet-induced MetS alters the remodeling enzyme LOX, thereby increasing the amount of crosslinking and inducing diastolic dysfunction.Furthermore we examined the role of T-lymphocytes in myocardial LOX regulation in diet-induced fibrotic hearts. Female SCID mice which are devoid of functional T-lymphocytes and C57BL/6 mice were treated with HFHSC diet for 12 months. Similar to male C67BL/6, female HFHSC-fed C57BL/6 mice demonstrated significant increase in maturation and catalytic activity of myocardial LOX, cross-linking, ventricular stiffness and diastolic dysfunction. Whereas induction of LOX protein was minimal in SCID mice compared with wild-type counterparts. Correspondingly fibrillar cross-linked collagen formation and diastolic dysfunction were less prominent in SCID mice. Our results suggest a potential role of T-lymphocytes in induction of myocardial stiffness and diastolic dysfunction through modulation of LOX-dependent collagen maturation.Moreover we studied the role of leptin, an adipokine over-produced in MetS with fibrotic effects in non-cardiac tissues, as a key mediator of profibrogenic responses in the heart by administrating leptin to C57BL/6 and leptin-deficient ob/ob mice. With exogenous leptin administration ob/ob mice displayed passive diastolic filling dysfunction that coincided with increase in myocardial collagen compared with ob/ob controls. Our findings suggest profibrotic effects of leptin in the heart, primarily through predominance of collagen synthesis over degradation.

Motivation: Cardiovascular dysfunction and hypertension are some of the leading causes of morbidity and mortality in the African population. According to the World Health Organisation the increases in these diseases are escalating in developing countries. Apart from the contributory role of genetics towards the incidence of hypertension, evidence regarding lifestyle as a determinant or marker of cardiovascular diseases in this group is not well known. The interaction of psychological and physiological mechanisms can contribute towards a broader scope of behavioural physiology in the higher prevalence of hypertension in Africans. Objectives: The main objective of the research in this thesis was to compare specific coping mechanisms of Africans with regard to cardiovascular dysfunction. Methodology: Manuscripts presented in Chapters 3, 4, and 5 made use of the cross-sectional comparative epidemiological "Transition and Health during Urbanisation in South Africa" (THUSA) project. The subjects included apparently healthy African men and women, which were recruited as a convenience sample from the North West Province, South Africa. Anthropometric measurements were taken and demographic questionnaires completed. An adapted Setswana COPE questionnaire was used to classify men and women as predominantly active (AC) or passive (PC) in coping style. Subjects were further subdivided into rural and urban groups (Manuscript Two), as well as younger (≤ 40) and older (≥ 45) age groups (Manuscript Three). The General Health Questionnaire (GHQ) was used to measure subjective perception of health in all three manuscripts. Blood pressure was recorded continuously before and during application of the handgrip test using the Finapres apparatus. Subjects were classified as normotensive and hypertensive after blood pressure measurement by the Finapres and the Riva-Rocci/Korotkoff method. The emphasis in this study was on the cardiovascular reactivity values. Fasting, resting serum renin activity, cortisol, prolactin, testosterone, high density lipoprotein, triglycerides, glucose and plasma fibrinogen values were correlated with cardiovascular and psychological variables. Significant differences between variables were determined by means of variance analyses (Manuscript One and Two adjusted for age; Manuscripts One, Two and Three adjusted for resting cardiovascular data). A logistic regression analysis was performed to determine the most significant determinants of urbanisation. All THUSA subjects and parents of under-aged adolescents gave informed consent and the study - was approved by the Ethics Committee of the Potchefstroom University for Christian Higher Education. The reader is referred to the abstracts at the beginning of each separate manuscript in Chapters 3 - 5 for a description of the subjects, study design and analytical methods used in each paper. Results and conclusions of the individual manuscripts: Results from the THUSA study showed that PC men and women reported more symptoms typical of an abnormal psychological and physiological profile than AC men and women. The PC men, compared to AC men, exhibited a larger vascular reactivity response as well as larger plasma renin activity. In contrast, the AC women showed a larger non-significant vascular reactivity response than PC women. All subjects though reacted with increased vascular reactivity on the stressor. Men with a PC strategy showed enhanced vascular reactivity, a perception of poorer health and larger stressor plasma renin activity. PC women reported more depressive symptoms and younger PC women indicated a higher prevalence of hypertension than younger AC women. As a follow-up on the first manuscript, the aim was focused mainly on including the environmental effect, namely urbanisation, as possible explanatory factor for the atypical physiological AC women’s' coping style. The rural AC subjects indicated more typical active coping central cardiac responses than rural PC subjects whereas urbanised AC and PC subjects indicated greater peripheral responses and hypertension prevalence rates. In addition, the urbanised AC men and women and PC women as opposed to their rural counterparts indicated symptoms more of a distress situation with increased values of prolactin and decreased values of testosterone. This was also accompanied by a perception of poorer health in women. Results of the AC style suggests that the typical physiological AC stimulation pattern of urbanised subjects and especially the women is dissociated from the "normal" physiological AC reaction and is now exhibited as a typical PC physiological stimulation pattern. The greater vascular reactivity, hypertension prevalence, perception of poorer health and endocrine distressed profile are associated with a PC and dissociated physiological AC style in an urban context in African men and women. No differences with regard to resting blood pressure or endocrine values were obtained when the AC and PC urbanised groups were compared. Africans develop cardiovascular dysfunction/hypertension during chronic stress or urbanisation. This implies a dissociation/habituation of physiological systems of African men and women despite having an active coping strategy. Active coping is, therefore, not necessarily "successful". Results of the first two manuscripts direct further investigation concerning the effects of ageing and urbanisation on the development of cardiovascular dysfunction and metabolic syndrome indicators in gender groups. The second manuscript showed that all rural AC subjects exhibit a more typical active coping central cardiac response and that rural PC and all urbanised subjects (AC and PC) exhibit enhanced peripheral vascular responses on the - handgrip test. Where peripheral vascular responses were more expected from older individuals in Manuscript Three, the occurrence of this pattern is strengthened in the younger subjects. The greater fibrinogen values in all younger urbanised women (AC and PC) compared to rural women further strengthen the risk for the development of cardiovascular disease. Increased vascular reactivity, abdominal obesity and increased levels of triglycerides as well as perception of poorer health were apparent in the urbanised AC women, PC men and women in comparison to their rural counterparts. The typical physiological AC stimulation pattern of urbanised women is dissociated from the "normal" physiological AC responses and is now exhibited as a typical PC physiological stimulation pattern. A typical PC style in older urbanised subjects is implicated in the greater hypertension prevalence. To conclude, it seems as if young urbanised Africans, and especially women, exhibit an AC style behaviourally with a dissociated physiological AC reaction pattern. Physiologically these women resemble a typical PC physiological cardiovascular and endocrine profile. This typical PC cardiovascular stimulation pattern is strengthened by a distressed endocrine profile, significant metabolic syndrome indicators and a 'perception of poorer health. Older PC style subjects also presented a greater hypertension prevalence. In this study it seems that cardiovascular changes that appear at a younger age might be influenced by other factors including urbanisation as a lifestyle factor as well as specific coping styles. Finally, a careful suggestion is made that specific coping mechanisms could be seen as a possible risk marker in the development of the metabolic syndrome. / Thesis (Ph.D. (Physiology))--North-West University, Potchefstroom Campus, 2005.