Vasospasm
BIBLIOGRAPHY
(1) Determine the tissue Selenium level of selected patients with fibroadenoma and cancer of the breast.
(2) Establish the similarity between the whole blood Selenium and tissue level in each of the groups of patients with fibroadenoma and cancer of the breast.
(3) Compare the whole blood Selenium level in the control group with the level in each of the groups with breast diseases.
(5) Further examine the relationship between selected predisposing factors and the tissue Selenium level.
3.2: STUDY DESIGN 3.2.1: Study Setting
The study was carried out at the Obafemi Awolowo University Teaching Hospitals Complex (OAUTHC), Ile-Ife, Osun State. The hospital has three General Surgical units which provide tertiary level services being mostly accessed by patients from Osun, Ondo, Ekiti and some part of Kwara, Kogi, Oyo and Edo-States with an estimated population of about 10 million according to 2006 National population census.
The people Ife/ Ijesa zone of Osun State where the institution is located are predominantly Yorubas with other Nigerian tribes and ethnic groups, however, also living permanently in the area. Peasant farming is the major occupation of the people. A sizeable number are engaged in commercial and small scale industrial enterprises, while the cream of the educated ones is the civil servants.
3.2.2: Methodology
3.2.2.1: Study participants
The study subjects consisted of three groups namely Group A – Control
Group B – Patients with fibroadenoma Group C – Patients with cancer
Subjects in the Control group were recruited from among the patients attending the General Surgery Out Patients Clinic for conditions other than breast diseases or cancer and who had no
previous or family history of breast diseases. Similarly, the controls were taken from the same catchment area that the patients with breast diseases reside.
For the fibroadenoma and cancer groups, consecutives patient undergoing operations for fibroadenoma and breast cancer conditions were recruited. They were formally enrolled only after histopathological examination of core needle or incisional or excisional biopsy had confirmed diagnosis.
The sociodemographic characteristics of all participants were recorded including age, gender, ethnicity, occupation, educational and socioeconomic status were recorded on a standard proforma.(Appendix I). The socioeconomic status was classified into high, middle and lower class using the educational level and occupation of the subjects and that of the parents when subjects were students as previously described by Oyedeji125 and modified by Ogunlesi et al 126. In the fibroadenoma and cancer groups the clinical information known to be relevant as predisposing factors were also recorded. Similarly, for the cancer group, tumor size, site of occurrence, stage at diagnosis, histopathological and immunohistochemical (IHC) status were documented.
Samples for histopathological examination were routinely processd in the laboratory using paraffin wax embedding and staining with Haematoxyllin and Eosin. Tissue processing for IHC staining was done according to standard protocol using monoclonal antibodies—1D5 clone for anti- ER, PR-2C5 for anti-PR and Z4881 for anti-HER2 (Zymed Laboratories, San Francisco, CA). Tumor blocks containing paraffin-embedded cell lines containing four cores—a negative CAL51 control cell line for orientation, MCF-7 ER positive cell line, T47D PR positive cell line and BT474 HER2 over-expressing cell lines (MaxArray Breast Receptor Control Cell Block—
evaluated for presence of positive reaction, cellular localization, pattern of staining and intensity of reaction. All slides were read by one pathologist and graded according to standard protocol into 0, 1+, 2+, 3+, using 10% positivity as the cutoff point for negative staining characteristics. 0 was classified as negative, 1+ and 2+ were classified as mild positive, 3+ and 4+ were classified as moderate positive while greater or equal to 5+ were regarded as strong positive.
Based on the IHC profile of the Estrogen receptors (ER), Progesterone receptors (PR) and Human epidermal growth factors receptors (HER), the cancer subjects were further classified into molecular subtypes: basal-like—triple negative—(ER-, PR-, HER2-), HER2+/ER- subtype (HER2+, ER-, PR-), luminal A (ER+ and/or PR+, HER2-), and luminal B (ER+and/or PR+, HER2+) as proposed by Adebamowo et al (2008)127.
3.2.2.1.1: Inclusion Criteria:
[1] Women with histologically confirmed breast cancer prior to primary surgery.
[2] Women with histologically confirmed fibroadenoma.
[3] Women with previous breast cancer presenting with local recurrence on ipsilateral and/or contra-lateral breast.
3.2.2.1.2: Exclusion Criteria:
[1] Patients who had been taking haematinics containing micronutrients, such as MIMS, within one week preceding presentation in the clinic.
[2] Patients with any gastrointestinal disease such as gastroenteritis and inflammatory bowel disease that can affect absorption of nutrients.
[3] Patients with previous history of gastric or intestinal surgery 3.2.2.2: Collection of blood and tissue samples
Five milliliters of blood sample was taken by venepuncture into an EDTA bottle from the fibroadenoma and cancer subject groups at the perioperative period. Blood samples for the determination of the whole blood selenium level were similarly taken from control subjects. The samples were stored at a refrigerator temperature of 40C until analyzed.
About one gram of tissue each was removed from the biopsy or the mastectomy specimens of patients with fibroadenoma and breast cancer and preserved in a specially prepared 10% formal saline till analysis was done. For mastectomy specimens, the tumor was bisected into half and the samples were taken from the core of the tissue so as to ensure that disease part was taken.
The limitation however exist that the sample part does not reflect the general tissue Selenium distribution due to the possibility of sampling necrotic tissue.