BIOCHEMICAL AND GENE EXPRESSION STUDIES ON UTERINE FIBROID PATIENTS IN IBADAN, OYO STATE, NIGERIA

Uterine fibroids (UF) are benign, non-carcinomatous growths that reside in the smooth layer of the uterus and cause many gynaecological problems among reproductive-age women worldwide. Their occurrence infringes on women's biological, sociological, physiological status and quality of lifestyle with an added financial burden, especially in black women. The actual cause of the phenomenon remains undiagnosed despite numerous studies, thereby prompting further investigation on its relationship to some biochemical and molecular markers of cellular growth and development. One hundred and ninety (190) reproductive-aged women (95 each for UF patients and age-matched non-UF participants) were recruited for the study. The experimental design was divided into two phases - Case-control model, and Case-control age-related model [16-25 years (group A), 26-35 years (group B), 36-45 years (group C), 46-55 years (group D) and 56 & above (group E)]. A portion of 5g each of the excised fibroid tissues and normal tissue samples from the adjacent myometrium were collected and sectioned into three parts. The first part was preserved using RNAlater® and kept at -80⁰ C for molecular analysis. The second portion was preserved with 10 % formalin solution for histological examination, while the last portion was rinsed with normal saline and frozen for biochemical analysis. The blood samples (10mL) were also collected; a 2mL portion was used for haematological analysis and the remaining 8mL for biochemical tests. Vitamins A, D, and K concentrations, the female reproductive hormones, and inflammatory markers (oestrogen, progesterone, IL-1, IL-2, CoX-1, CoX-2, and TNF-α respectively) were assessed using Enzyme-linked immunosorbent assay kits. Concentrations of mineral elements in UF and non-UF were examined using atomic absorption spectrophotometry. The supernatant obtained from tissues and serum was utilised to assess the antioxidant status, and vitamins E and C concentrations were investigated spectrophotometrically. Age, reproductive hormones, antioxidant enzymes and micronutrients were identified as contributing factors promoting the incidence or occurrence of the UF. The micronutrient examination in the phase one design model revealed that selenium of all the examined mineral elements and vitamins A, D, and E were significantly reduced while Vitamin K increased significantly in UF cases at (p<0.05) promoting the occurrence of tumour. There was a significant increase (p<0.05) in the level of vitamin K, sodium, magnesium, iron, zinc and IL-2 in group C; Ca in group E, progesterone in groups B and C, reduction of progesterone in group D, estrogen, and cholesterol in group D; the activities of glutathione-s-transferase and over expression of transforming growth factor gene in the fibroid tissues. Conversely, there was a significant reduction of (P<0.05) in the levels of reduced glutathione in groups B, C and E, and SOD and selenium in groups E and D respectively. Similarly, there was a significant (P<0.05) decrease in the levels of vitamin A in groups B and C, vitamin D in groups D, vitamin E in groups D and E, sodium and cholesterol in group B in UF cases. The UF incidence or occurrence were associated with the nutritional influence of vitamin E, A = D, and selenium deficiencies in decreasing order, excess vitamin K intake and with overexpression of TGF-β. Thus, biochemical and molecular processes related to dietary lifestyle are strongly associated factors facilitating the occurrence of UF in most Black women

Evaluation of Goitrogenic Content of Common Vegetables in South West Nigeria

Endemic goiter and associated iodine deficiency disorders (IDD) are prevalent in south west Nigeria. The present study was undertaken to identify the role of dietary goitrogen in the etiology of endemic goiter. Perchlorate, fluoride, nitrate, bromide, chloride, phosphate and cyanide content of common vegetables viz., cabbage, African Eggplant, Giant pigweed, Scent leaf, Amaranth, Tree Spinach, Black nightshade, consumed by the population of the region were measured. All the dietary goitrogen content in the investigated vegetables were found to be within the Acceptable Daily Intake and this observation suggests that in addition to iodine deficiency dietary intake of a cyanogenic plant, the combination of chemicals may play some role for the persistence of endemic goiter in Southwest Nigeria

Ameliorative Effect of Zingiber officinale on Chemical Induced DNA Damage in Rats Using PCR Analysis

Ginger rhizomes have been reportedly used in folk medicine for the management of various ailments. This study, therefore, investigates the ameliorative effect of the ethanolic extract of ginger (Zingiber officinale) rhizomes against DNA damage in rats induced with different carcinogens. Fifteen Wistar rats grouped into 3 of 5 rats per group were used for the study. The first set of blood samples was first collected before the animals were orally treated with heavy metals. After 14 days of induction, the second set of blood was collected. The third phase of blood collection was done after administering an ethanolic extract of Z. officinale for 14 days. The UV wavelength absorption spectrum and conventional PCR analysis were carried out on DNA extracts of all the animals. Cluster analysis of optical density (OD) and PCR data were carried out as well as genomic instability, similarity, and diversity using the best 3 Random Amplified Polymorphic DNA (RAPD) primers. The PCR –DNA concentration analysis showed the Z. officinale extract's ameliorative effect against lead acetate, cadmium chloride, and arsenic trioxide-induced DNA damage with a significant (p < 0.05) reduction in DNA concentration of the treated rats when compared with induced rats. The cluster analysis of optical density values revealed close similarity between the control animals' DNA, a slight similarity with treated animals' DNA, and a significant difference with the induced animal DNA. These results indicated the ameliorative properties of Z. officinale against these heavy metals induced DNA damage in rats

Macrophages are exploited from an innate wound healing response to facilitate cancer metastasis.

Tumour-associated macrophages (TAMs) play an important role in tumour progression, which is facilitated by their ability to respond to environmental cues. Here we report, using murine models of breast cancer, that TAMs expressing fibroblast activation protein alpha (FAP) and haem oxygenase-1 (HO-1), which are also found in human breast cancer, represent a macrophage phenotype similar to that observed during the wound healing response. Importantly, the expression of a wound-like cytokine response within the tumour is clinically associated with poor prognosis in a variety of cancers. We show that co-expression of FAP and HO-1 in macrophages results from an innate early regenerative response driven by IL-6, which both directly regulates HO-1 expression and licenses FAP expression in a skin-like collagen-rich environment. We show that tumours can exploit this response to facilitate transendothelial migration and metastatic spread of the disease, which can be pharmacologically targeted using a clinically relevant HO-1 inhibitor

Evaluation of Goitrogenic Content of Common Vegetables in South West Nigeria

Endemic goiter and associated iodine deficiency disorders (IDD) are prevalent in south west Nigeria. The present study was undertaken to identify the role of dietary goitrogen in the etiology of endemic goiter. Perchlorate, fluoride, nitrate, bromide, chloride, phosphate and cyanide content of common vegetables viz., cabbage, African Eggplant, Giant pigweed, Scent leaf, Amaranth, Tree Spinach, Black nightshade, consumed by the population of the region were measured. All the dietary goitrogen content in the investigated vegetables were found to be within the Acceptable Daily Intake and this observation suggests that in addition to iodine deficiency dietary intake of a cyanogenic plant, the combination of chemicals may play some role for the persistence of endemic goiter in Southwest Nigeria

Ethyl acetate leaf fraction of Cnidoscolus aconitifolius (Mill.) I. M. Johnst: antioxidant potential, inhibitory activities of key enzymes on carbohydrate metabolism, cholinergic, monoaminergic, purinergic, and chemical fingerprinting

In this study, the antioxidant, carbohydrate metabolism, cholinergic, monoaminergic, and purinergic enzyme activities of ethyl acetate leaf fraction of Cnidoscolus aconitifolius were evaluated with the high-performance liquid chromatography (HPLC) analysis. The ethyl acetate fraction of the plant leaf was tested for antioxidant properties, key carbohydrate metabolism, cholinergic, monoaminergic, and purinergic enzyme activities using standard procedures, while the chemical composition of the fraction was evaluated using HPLC. The results revealed that the fraction has higher phenolic compounds than flavonoid and exhibited the ability to scavenge iron chelation and ABTS. The fraction also inhibited the activities of α-amylase and α-glucosidase, acetylcholinesterase, butyrylcholinesterase, monoamine oxidase, tyrosinase, arginase, Ecto-5’-nucleotidase, phosphodieterase-5, angiotensin-I-converting enzyme and encouraged the activity of Na+/K+-ATPase. The HPLC analysis revealed that the ethyl acetate fraction contained coumaric acid, amentoflavone, hesperidin, protocatechuic acid, kaempferol, dihydromyricetin, quercetin,and rutin. The obtained results in this study suggest that the ethyl acetate fraction obtained from aqueous extracts of Cnidoscolus aconitifolius leaf possessed outstanding antioxidative potentials. Thus, excellent enzyme inhibitory activities probably due to bioactive compounds are also observed in the leaf

A quantitative approach for determining the role of geometrical constraints when shaping mesenchymal condensations

In embryogenesis, mesenchymal condensation is a critical event during the formation of many organ systems, including cartilage and bone. During organ formation, mesenchymal cells aggregate and undergo compaction while activating developmental programmes. The final three-dimensional form of the organ, as well as cell fates, can be influenced by the size and shape of the forming condensation. This process is hypothesized to result from multiscale cell interactions within mesenchymal microenvironments; however, these are complex to investigate in vivo. Three-dimensional in vitro models that recapitulate key phenotypes can contribute to our understanding of the microenvironment interactions regulating this fundamental developmental process. Here we devise such models by using image analysis to guide the design of polydimethylsiloxane 3D microstructures as cell culture substrates. These microstructures establish geometrically constrained micromass cultures of mouse embryonic skeletal progenitor cells which influence the development of condensations. We first identify key phenotypes differentiating face and limb bud micromass cultures by linear discriminant analysis of the shape descriptors for condensation morphology, which are used to guide the rational design of a micropatterned polydimethylsiloxane substrate. High-content imaging analysis highlights that the geometry of themicroenvironment affects the establishment and growth of condensations. Further, cells commit to establish condensations within the first 5 h; condensations reach their full size within 17 h; following which they increase cell density while maintaining size for at least 7 days. These findings elucidate the value of our model in dissecting key aspects of mesenchymal condensation development