Expression Of Apoptosis Markers In The Skin Microvasculature Of Patients With Diabetes Mellitus Type 2 At Hospital Universiti Sains Malaysia [QH671. D161 2008 f rb].

Perubahan salur darah pada pesakit diabetes jenis 2 berkait rapat dengan kesan jangka panjang komplikasi diabetes. The microvascular changes in diabetes are directly related to the long term complications of diabetes

Elucidating the effects of rapamycin and PF4 on the MNU induced female rats and human breast cancer cells (MCF-7)

There is a strong evidence that tumour growth is not just a consequence of uncontrolled proliferation but also of reduced apoptosis. Bax and Bcl-2 are Bcl-2 family like apoptosis regulator. They function either as suppressor (Bcl-2) or a promoter (Bax) of apoptosis. As a member of the inhibitors of apoptosis (IAP), survivin is also a promoter of cellular proliferation and thus a key player in cancer progression. Beside, caspases also occupy a central role in maintaining cellular homeostasis. Therefore, this research aims to examine the apoptotic effects of Rapamycin and Platelet Factor 4 (PF4) on 1-methyl-1-nitrosourea (MNU)-induced mammary carcinoma through Bcl-2- survivin and caspase modulated pathways in female Sprague Dawley Rat (SDR) and in vitro MCF-7 breast cancer cell lines. One hundred 21 days old female SDR were given an intraperitoneal injection (IP) of MNU to induce breast cancer formation. When tumour size reached 14.5±0.5mm, intratumoural injections of the following interventions were given; Group 1 (preintervention control, n=20 and post-intervention control, n=20), Group 2 (Rapamycintreated, n=20), Group 3 (PF4-treated, n=20) and Group 4 (Rapamycin+PF4-treated, n=20). Tumour growth was then morphologically assessed using haematoxylin and eosin (H&E) and immunohistochemistry (IHC) utilizing pro-apoptosis (Bax) and antiapoptosis markers (Bcl-2), survivin and caspases-3,-6,-7,-8, and -9. Besides that, the MCF-7 cell line was used for in vitro assessment. Initially, half maximal inhibitory concentration (IC50) of each drug was determined. The MCF-7 cell lines were then exposed to Rapamycin and PF4 and Rapammycin+PF4 at IC50 concentrations, after which they were subjected to flow cytometry and Western blot analyses. Bax was significantly expressed at higher levels in the rapamycin-treated and Rapamycin+PF4- treated groups than controls (p<0.001). Besides, survivin was significantly downregulated in the PF4-treated and Rapamycin+PF4-treated group when compared to controls (p<0.001). On the other hand, Bcl-2 expression was found not to be significantly altred in all treatment groups. Caspase-3, was significantly expressed at higher levels in both PF4-treated and Rapamycin+PF4-treated groups than controls (p<0.001), as well as rapamycin-only group when compared to Rapamycin+PF4- treated and PF4-treated groups (both p<0.001). Apart from that, caspase-6 was also significantly expressed in Rapamycin-treated, PF4-treated and Rapamycin+PF4- treated than the control groups (p<0.001). Besides, either PF4 or Rapamycin+PF4 combination was associated with increased caspase-7 expression, compared to the controls (p<0.001). However, Rapamycin-treated group showed a significantly higher caspase-8 expression when compared to PF4-treated (p<0.05). For caspase-9, higher caspase-9 expression was observed in Rapamycin-treated group when compared to control, Rapamycin+PF4-treated and PF4-treated cohorts (p<0.001). The IC50 Rapamycin, PF4 and Rapamycin+PF4 were 0.4 μg/ml, 6 μg/ml and 0.4 μg/ml+1.0 μg/ml respectively. Rapamycin and PF4, on the other hand, were non-toxic to the normal HMEC cells. Furthermore, Rapamycin, PF4 and Rapamycin+PF4 induced a cell cycle arrest in MCF-7 cell lines at G0/G1, S and G0/G1 phases, respectively. Rapamycin, PF4 and Rapamycin+PF4 induced the upregulation of pro-apoptotic Bax and the downregulation of anti-apoptotic Bcl-2 and survivin. The expression levels of caspase-3 and caspase-8 were consistent in the treated group when compared with the control group. The expression of caspases -6, -7 and -9 protein were increased when treated with Rapamycin, Rapamycin + PF4 and PF4 when compared with the control group. In conclusion, this study provides new insights on the mechanistic properties of Rapamycin and PF4 as anti-cancer agents in breast cancer animal model and in vitro MCF-7 cell lines. The results lend support to the notion that apoptotic induction by Rapamycin and PF4 and was mediated by both the intrinsic and extrinsic pathways through the activation of caspase-9, caspase-3 and caspase-8 activation

Association of serum vitamin D level and vitamin D receptor expression among newly diagnosed breast cancer

There is a mushrooming interest in the anti-carcinogenic property of vitamin D. However, many researches reported a conflicting result in the association of vitamin D levels to certain types of cancer. This study was designed to assess the association between vitamin D and vitamin D receptor (VDR) expression with breast cancer. This case-control study, carried out at Hospital Universiti Sains Malaysia, Kelantan, involved 69 newly diagnosed breast cancer patients and 73 healthy volunteers. Serum 25(OH)D was taken and compared between 2 groups. VDR expression in patients’ breast tissue samples was determined by immunohistochemical staining method using anti-VDR antibody. 85.5% of breast cancer patients and 97.3% of healthy control were vitamin D insufficient with a mean (SD) of 13.36 (6.96) ng/mL and 13.05 (3.71) ng/mL, respectively, and the difference was not statistically significant. VDR expression showed cytoplasmic positivity in 75.4% of breast cancer tissue, followed by both cytoplasmic and nuclear positivity in 21.5% and complete absence in 3%. There was no significant association between VDR expression and hormone receptor status. In conclusion, there was a high prevalence of vitamin D deficiency among breast cancer and healthy volunteers in our study. There was no significant association between breast cancer and vitamin D. The VDR expression in breast cancer cells showed high cytoplasmic localization

A panel of three microRNA signatures as a potential biomarker for CRC screening based on stages and functional prediction using bioinformatic analysis

Background: MicroRNA (miRNA) has been linked to colorectal cancer (CRC) tumorigenesis due to its post-transcriptional mechanism in targeting cancer-associated genes. Although miRNAs appear to be promising screening biomarkers, functional prediction analysis is required to shed light on their role in CRC tumorigenesis. Therefore, this study aims to identify the significantly deregulated miRNAs in CRC tumorigenesis. (2) Methods: Three upregulated miRNAs (hsa-miR-20a-5p, hsa-miR-21-5p, and hsa-miR210-3p) from 14 significant differentially expressed miRNAs (DEMs) were chosen from microarray profiling to be validated in plasma. Bioinformatics analyses showed that these miRNAs generally contributed to tumorigenesis, but only hsa-miR-20a-5p and hsa-miR-21-5p were specifically linked to CRC. Only two miRNAs showed a positive correlation when compared to their expression in plasma. However, further analysis showed that all three miRNAs in plasma were significantly difference between the early and advanced stages of CRC. ROC curve analysis was used to evaluate miRNAs’ diagnostic performance in the early and advanced stages. (3) Results: Collectively, hsa-miR-20a-5p showed the highest discriminative value (AUC= 0.82, sensitivity = 86%, and specificity= 88%) in discriminating early CRC, while both hsa-miR-21-5p and hsa-miR-210-3p give a perfect performance for advance CRC. In addition, the performance of all miRNAs’ combinations also gives a perfect performance for diagnosis in both early and advanced CRC, except the combination of hsa-miR-20a-5p and hsa-miR-210-3p. (4) Conclusions: A few potential miRNA panels as CRC biomarker is needed for better prediction of disease. The reflective circulating miRNAs can be contributed to by minimal invasive screening tools

Novel Drug and Gene Delivery System and Imaging Agent Based on Marine Diatom Biosilica Nanoparticles

Mesoporous silica nanoparticles (MSNs) have great potential for applications as a drug delivery system (DDS) due to their unique properties such as large pore size, high surface area, biocompatibility, biodegradability, and stable aqueous dispersion. The MSN-mediated DDS can carry chemotherapeutic agents, optical sensors, photothermal agents, short interfering RNA (siRNA), and gene therapeutic agents. The MSN-assisted imaging techniques are applicable in cancer diagnosis. However, their synthesis via a chemical route requires toxic chemicals and is challenging, time-consuming, and energy-intensive, making the process expensive and non-viable. Fortunately, nature has provided a viable alternative material in the form of biosilica from marine resources. In this review, the applications of biosilica nanoparticles synthesized from marine diatoms in the field of drug delivery, biosensing, imaging agents, and regenerative medicine, are highlighted. Insights into the use of biosilica in the field of DDSs are elaborated, with a focus on different strategies to improve the physico-chemical properties with regards to drug loading and release efficiency, targeted delivery, and site-specific binding capacity by surface functionalization. The limitations, as well as the future scope to develop them as potential drug delivery vehicles and imaging agents, in the overall therapeutic management, are discussed

The Unique Biology behind the Early Onset of Breast Cancer

Breast cancer commonly affects women of older age; however, in developing countries, up to 20% of breast cancer cases present in young women (younger than 40 years as defined by oncology literature). Breast cancer in young women is often defined to be aggressive in nature, usually of high histological grade at the time of diagnosis and negative for endocrine receptors with poor overall survival rate. Several researchers have attributed this aggressive nature to a hidden unique biology. However, findings in this aspect remain controversial. Thus, in this article, we aimed to review published work addressing somatic mutations, chromosome copy number variants, single nucleotide polymorphisms, differential gene expression, microRNAs and gene methylation profile of early-onset breast cancer, as well as its altered pathways resulting from those aberrations. Distinct biology behind early-onset of breast cancer was clear among estrogen receptor-positive and sporadic cases. However, further research is needed to determine and validate specific novel markers, which may help in customizing therapy for this group of patients

Comparing the Biology of Young versus Old Age Estrogen-Receptor-Positive Breast Cancer through Gene and Protein Expression Analyses

Background: Breast cancer developed at a young age (≤45 years) is hypothesized to have unique biology; however, findings in this field are controversial. Methods: We compared the whole transcriptomic profile of young vs. old-age breast cancer using DNA microarray. RNA was extracted from 13 fresh estrogen receptor (ER)-positive primary breast cancer tissues of untreated patients (7 = young age ≤45 years and 6 = old age ≥55 years). In silico validation for the differentially expressed genes (DEGs) by young-age patients was conducted using The Cancer Genome Atlas (TCGA) database. Next, we analyzed the protein expression encoded by two of the significantly down-regulated genes by young-age patients, Glycine N-acyltransferase-like 1 (GLYATL-1) and Ran-binding protein 3 like (RANBP3L), using immunohistochemical analysis in an independent cohort of 56 and 74 ER-positive pre-therapeutic primary breast cancer tissues, respectively. Results: 12 genes were significantly differentially expressed by young-age breast cancers (fold change >2 or p-value p-value 0.005) associated with positive lymph node status. Higher expression of RANBP3L was significantly associated with breast cancers with lower histopathological grades (p-value 0.038). Conclusions: At the transcriptomic level, breast cancer developed in young and old age patients seems homogenous. The variation in the transcriptomic profiles can be attributed to the other clinicopathological characteristics rather than the age of the patient

Comparing the Biology of Young versus Old Age Estrogen-Receptor-Positive Breast Cancer through Gene and Protein Expression Analyses

Background: Breast cancer developed at a young age (≤45 years) is hypothesized to have unique biology; however, findings in this field are controversial. Methods: We compared the whole transcriptomic profile of young vs. old-age breast cancer using DNA microarray. RNA was extracted from 13 fresh estrogen receptor (ER)-positive primary breast cancer tissues of untreated patients (7 = young age ≤45 years and 6 = old age ≥55 years). In silico validation for the differentially expressed genes (DEGs) by young-age patients was conducted using The Cancer Genome Atlas (TCGA) database. Next, we analyzed the protein expression encoded by two of the significantly down-regulated genes by young-age patients, Glycine N-acyltransferase-like 1 (GLYATL-1) and Ran-binding protein 3 like (RANBP3L), using immunohistochemical analysis in an independent cohort of 56 and 74 ER-positive pre-therapeutic primary breast cancer tissues, respectively. Results: 12 genes were significantly differentially expressed by young-age breast cancers (fold change >2 or <2- with FDR p-value < 0.05). TCGA data confirmed the differential expression of six genes. Protein expression analysis of GLYATL-1 and RANBP3L did not show heterogeneous expression between young and old-age breast cancer tissues. Loss of expression of GLYATL-1 was significantly (p-value 0.005) associated with positive lymph node status. Higher expression of RANBP3L was significantly associated with breast cancers with lower histopathological grades (p-value 0.038). Conclusions: At the transcriptomic level, breast cancer developed in young and old age patients seems homogenous. The variation in the transcriptomic profiles can be attributed to the other clinicopathological characteristics rather than the age of the patient.- Ministry of Higher Education Malaysia (MOHE) - the fundamental research grant scheme (FRGS), grant # [FRGS/1/2019/SKK08/USM/02/15]

A preliminary study on the effects of stingless bee honey (SBH) on fasting blood glucose in streptozotocin (STZ)-induced diabetic rat models

Diabetes is a metabolic disorder characterized by hyperglycemia due to defects in insulin secretion, insulin action, or both. SBH has good antihyperglycemic potential and has traditionally been used as an alternative treatment for diabetes mellitus (DM). Since the role of SBH in glucose control is still unclear in animal and human studies, the present study was designed to evaluate the antihyperglycemic effects of SBH in streptozotocin-induced diabetic rat models. Fifteen Sprague-Dawley rats (200-250 g) were equally divided into five groups, the first group being a normal (non-diabetic) rat and the other four groups being diabetic. The normal and untreated diabetic groups received normal saline while the other diabetic groups were treated with SBH (2 g/kg body weight), metformin (MET /250 mg/kg body weight) and SBH + MET respectively. The treatment was given within 12 days. Fasting blood glucose (FBG) was measured at baseline and every two weeks thereafter. On days 7 and 12, SBH significantly lowered FBG, comparable to the normal group (p<0.05). In the group treated with MET and the combination of SBH-MET, FBG improved only on the 12th day of treatment (p<0.05). The results show that a single SBH treatment is effective in lowering blood glucose levels. Thus, SBH could be of great value in the treatment of diabetes in humans

A preliminary study on the effects of Stingless Bee Honey (SBH) on fasting blood glucose in Streptozotocin (STZ)-Induced diabetic rat models

Diabetes is a metabolic disorder characterized by hyperglycemia due to defects in insulin secretion, insulin action, or both. SBH has good antihyperglycemic potential and has traditionally been used as an alternative treatment for diabetes mellitus (DM). Since the role of SBH in glucose control is still unclear in animal and human studies, the present study was designed to evaluate the antihyperglycemic effects of SBH in streptozotocin-induced diabetic rat models. Fifteen Sprague-Dawley rats (200-250 g) were equally divided into five groups, the first group being a normal (non-diabetic) rat and the other four groups being diabetic. The normal and untreated diabetic groups received normal saline while the other diabetic groups were treated with SBH (2 g/kg body weight), metformin (MET /250 mg/kg body weight) and SBH + MET respectively.The treatment was given within 12 days. Fasting blood glucose (FBG) was measured at baseline and every two weeks thereafter. On days 7 and 12, SBH significantly lowered FBG, comparable to the normal group (p<0.05). In the group treated with MET and the combination of SBH-MET, FBG improved only on the 12th day of treatment (p<0.05). The results show that a single SBH treatment is effective in lowering blood glucose levels. Thus, SBH could be of great value in the treatment of diabetes in human