Insulin-like Growth Factor-1 and Calcium Ion Levels are Negatively Associated with Serum β-Cross Laps in Multi-transfused β-thalassemia Major Patients

BACKGROUND: Thalassemia patients with repeated transfusions (multi-transfusion) are often at risk of experiencing early osteoporosis. Several studies have demonstrated that osteoporosis in these individuals was associated with altered bone remodeling, characterized by decreased insulin-like growth factor-1 (IGF-1) and serum calcium ions, as well as increased serum β-Cross Laps (β-CTx) levels. Despite the prevalence of this condition, there is limited literature on the relationship between IGF-1 levels and calcium ions with β-CTx. Therefore, this study was conducted to examine the relationship between IGF-1 and calcium ions levels with serum β-CTx in multi-transfused β-thalassemia major patients.METHODS: A cross sectional study involved 29 thalassemia patients with multiple transfusions, aged 2-18 years, that were selected from the electronic medical records. Calcium ion levels were examined using ion selective electrode method. Subsequently, IGF-1 and serum β-CTx levels were examined by enzyme-linked immunosorbent assay (ELISA), and the data were analyzed using the Pearson correlation test.RESULTS: The results showed that the mean serum IGF-1 levels, calcium ion, and β-CTx were 20.11±20.763 ng/mL, 1.26±0.07 mmol/L, and 9330.40±1696.76 ng/mL, respectively. Statistical analysis showed a significant relationship between the levels of IGF-1 (r=-0.573; p=0.001) and calcium ion (r=-0.373; p=0.046) with serum β-CTx. A moderate negative relationship was found between IGF-1 levels and β-CTx, while calcium ion levels and β-CTx showed a weak negative relationship.CONCLUSION: A moderate negative correlation between IGF-1 and serum β-CTx, and a weak negative correlation between calcium ion and serum β-CTx suggest that IGF-1 and calcium ions may serve as potential indicators of bone turnover and osteoporosis risk in multi-transfused β-thalassemia major patients, underscoring their potential role in routine clinical evaluations.KEYWORDS: miR-200a expression, NO level, early-onset preeclampsia, late-onset preeclampsi

Curcumin Enhances Antimigration of Pentagamavunon-1 by Suppressing MMP-2 and MMP-9 Expression in Triple-Negative (4T1) and Luminal A (T47D) Breast Cancer Cells

BACKGROUND: Breast cancer remains the leading cause of cancer death worldwide. Migration and invasion of cancer cells are still crucial stages in the metastasis process, highlighting the urgent need for treatments that target both proliferation and metastatic progression. Curcumin and its synthetic analogue, pentagamavunon (PGV)-1, exhibit antiproliferative effects in breast cancer cells. However, the effects of combining curcumin and PGV-1 on cancer cell migration have not yet been explored. Therefore, this study was conducted to examine the antimigratory effects of curcumin and PGV-1 combination on 4T1 and T47D breast cancer cells.METHODS: Cytotoxicity effects of curcumin and PGV-1 were examined using an MTT assay to determine their effects on 4T1 and T47D cell viability. The antimigration activity was assessed using a scratch wound healing assay by measuring the closure of artificially created wounds on monolayer cells. Expression of matrix metalloproteinases (MMPs) that play a crucial role in cancer cell migration was analyzed using gelatin zymography to measure their enzymatic activities.RESULTS: The IC50 of PGV-1 and curcumin were 4.88 μM and 37.62 μM in 4T1 cells and 3.16 μM and 23.15 μM in T47D cells, respectively. Furthermore, combination of PGV-1 and curcumin effectively inhibited 4T1 and T47D cell migration. PGV-1 (0.5–2 μM) demonstrated superior antimigratory activity compared to curcumin (5–20 μM) by suppressing MMP-2 and MMP-9 expression in both cell lines. Significantly, curcumin was shown to synergistically enhance the antimigratory effects of PGV-1, leading to a further decrease in MMP-2 and MMP-9 expression. CONCLUSION: The combination of PGV-1 and curcumin may provide a promising antimigratory agent, potentially leading to enhanced antimetastatic strategies and more efficacious treatments for triple-negative and luminal breast cancer patients.KEYWORDS: antimigration, curcumin, luminal breast cancer, MMP-2, MMP-9, pentagamavunon-1, triple-negative breast cance

HPV18 E6/E7 Mutation and Their Association with The Expression Level of Tumor Suppressor Proteins p53 and pRb among Indonesian Women with Cervical Cancer

BACKGROUND: The E6/E7 mutation contributes to the intra-typic variant of HPV18 which may differ in their oncogenic potential. E6 and E7 target the tumour suppressor protein p53 and pRb, respectively, and their degradation play a crucial role in cervical carcinogenesis. However, the prevalence of HPV18 E6/E7 variants among Indonesian women with cervical cancer has not been elucidated. Therefore, this study was conducted to characterize the HPV18 variants among Indonesian women with cervical cancer and their association with tumor suppressor protein p53 and pRb.METHODS: A hundred Indonesian women with pathologically proven cervical cancer were consecutively recruited into the study. Polymerase chain reaction (PCR) was performed to detect HPV18 DNA E6 and E7 oncogenes using specific primers and the variants was determined through nucleotide sequencing. Expressions of p53 and pRb were analyzed through immunohistochemistry by using specific antibodies targeting p53 and pRB.RESULTS: The rate of HPV18 positivity was 24%. The rate of E6 and E7 mutation was 45.4% and 59.1%, respectively. Those with E6 mutation had significantly higher expression of p53 and pRb as compared to those with wildtype E6 (p<0.05). Subjects with E7 mutation only had higher expression of pRb (p<0.05). Phylogenetic analysis revealed that 54.5% subjects had genetic sequences closely related to Asian lineages, particularly A1, A4, and A5 sublineage. Interestingly, 3 subjects had genetic sequences closely related to MK813921, a newly identified sequences. However, 45.5% subjects had distinct genetic sequences that did not related to the reference sequence used in this study.CONCLUSION: E6 and E7 mutation was common among Indonesian women with HPV18 cervical cancer and associated with the level of tissue p53 and pRb expression.KEYWORDS: HPV18 E6/E7, mutation, epidemiology, Indonesian wome

Safety Concerns of Tectona grandis L.f. Leaf Extract as a Natural Food Colorant: Evidence of Irreversible Organ Pathology in Subchronic Toxicity Study

BACKGROUND: Tectona grandis Lf (TG) leaves are traditionally used in Indonesia for natural dyeing, and are gaining popularity as food colorants globally. However, their safety profile remains unclear. Acute toxicity studies reported no fatalities at doses up to 5000 mg/kg BW, histological analyses revealed inflammation and necrosis in the stomach, raising concerns regarding the long-term safety of TG leaf extract. Therefore, this study was performed to evaluate subchronic toxicity of TG leaf extract in both males and females Wistar rats.METHODS: TG leaf extracts were obtained by water extraction and extract powder was suspended in sodium carboxymethyl cellulose (Na-CMC). Male and female rats were administered TG leaf extract at doses of 0, 10, 20, and 40 mg/kg BW for 28 days, with a 14-days recovery phase in the satellite groups (as controls). Hematology profiles and biochemistry were analyzed using hematology analyzer and spectrophotometry. Histology analysis was performed to investigate TG effects on the organs.RESULTS: Hematological analysis revealed reversible reductions in hemoglobin, erythrocyte, and hematocrit levels, along with irreversible decreases in leukocyte and thrombocyte. While TG leaf extract did not significantly affect serum glutamic pyruvic transaminase (SGPT), serum glutamic oxaloacetic transaminase (SGOT), or creatinine levels, bilirubin levels increased, but remained within normal ranges. However, histopathological findings at 40 mg/kg BW revealed congestion and inflammation in the digestive organs, as well as neutrophil infiltration and congestion in metabolism-related organs, the lungs, liver, kidneys, and lymph nodes. These pathological changes persisted throughout the recovery period.CONCLUSION: TG leaf extract raises safety concerns, particularly at a dose of 40 mg/kg BW, as it induces irreversible organ pathology despite reversible changes in blood parameters.KEYWORDS: Indonesian Tectona grandis Lf, subchronic, toxicity, natural, food, colorant

Transcriptional Regulation of CYP2D6 by Nrf2 and Its Implications in Breast Cancer Therapy: Bioinformatics and Experimental Evidence

BACKGROUND: Tamoxifen (TAM) resistance in patient with breast cancer is the leading cause of mortality among women globally. Cytochrome P450 2D6 (CYP2D6) is involved in the metabolism of TAM, and recently NF-E2-related factor 2 (Nrf2) has recently been found as its regulator. However, the impact of Nrf2-mediated CYP2D6 regulation in the context of breast cancer and TAM resistance are currently unknown. Therefore, this study was conducted to examine the role of CYP2D6 and Nrf2 in breast cancer prognosis. MEDTHODS: The roles of CYP2D6 and Nrf2 were investigated in the T47D breast cancer cell line and T47D-derived TAM-resistant cells by examining the gene expression, cell viability, and transcriptional regulation by quantitative reverse transcription polymerase chain reaction (qRT-PCR), MTT, and reporter gene assay, respectively. Additionally, comprehensive in silico analysis of the transcriptomic and clinical data from The Cancer Genome Atlas database were performed to uncover the prognostic role of CYP2D6 and its regulator in breast cancer patients. RESULTS: CYP2D6 mRNA was low and Nrf2 protein was high in TAM-resistant T47D cells compared to parental cells. Nrf2 knockdown upregulated CYP2D6 mRNA levels and enhanced the cytotoxicity of TAM. Similarly, in silico analysis revealed that low CYP2D6 mRNA and high Nrf2 protein were related to a lower probability of survival. The rs1238662089 within the identified Nrf2-binding site was found to greatly affect CYP2D6 expression levels, indicating its role as predictor for better prognosis. CONCLUSION: This study revealed for the first time that Nrf2 regulates CYP2D6expression in breast cancer and is involved in TAM sensitivity; thus, plays a role in breast cancer patient prognosis.KEYWORDS: breast cancer, CYP2D6, Nrf2, pharmacoepigenetics, SNP

Resveratrol: The Multifaceted Roles and Mechanisms of Polyphenol to Improve Longevity, Immunomodulation, and Age-related Diseases

High in polyphenols diet has been known to protect human against chronic metabolic diseases including cancer, diabetes, neurological and cardiovascular disorders. Resveratrol (RSV) is a natural polyphenol that presents in fruits, vegetables, and nuts. The polyphenols content of RSV possesses anti-inflammatory, antioxidant, immunomodulatory, and anticancer properties by influencing the nuclear factor-kappaB (NF-κB), p53, adenosine monophosphate-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR), Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathways, enzymatic antioxidants expressions, and the levels of microRNAs. Therefore, this review article will focus on the potential of RSV in improving aging and metabolic diseases, which mostly induced by low-chronic inflammation and oxidative stress. RSV is also known as calorie restriction (CR)-mimetics to activate sirtuins family which improve mitochondrial function, repair DNA and genomic stability and reduce inflammation thus become a promising substance to extend health span and longevity. RSV can be useful as a supplement to prevent aging-related diseases, with a dose range between 250–1,000 mg depending on the intended health benefit and individual factors. More clinical data is needed to determine the impact of RSV metabolites and the relationship between dose, concentration, and effect, particularly in the context of chronic illness.KEYWORDS: mesenchymal stem cell, extracellular vesicle, exosome, cancer therapy, drug deliver

Granule Nanoparticle Citrus sinensis (L.) Osbeck Peel Lowers Blood Glucose Levels and HbA1C in Alloxan-induced Diabetes Rats

BACKGROUND: The peel of Citrus sinensis (L.) Osbeck (sunkist orange) peels, which are often seen as waste, actually contains valuable properties such as antioxidants, hypoglycemic, nephroprotective, and anti-inflammatories. The potential effects of C. sinensis peel on diabetes have been discussed but not clear yet. Therefore, this study was performed to evaluate the effects of Granule Nanoparticle Sunkist Peel (GNSP) extracts as an antidiabetic agent in alloxan-induced diabetic rats.METHODS: The nanoparticle suspension was prepared by mixing a formulation of 0.2% chitosan and 0.1% sodium tripolyphosphate. The characteristics of nanoparticles were measured by flow time, tap index and angle of repose. Rats were induced with alloxan injection to create diabetes rat models. Rats were divided into five groups; normal control group, diabetic controls, and diabetic rats receiving either 50, 100, or 200 mg/kg/day GNSP. After 28 days of diabetes induction, rats were euthanized, and blood as well as tissue samples were collected. Blood glucose levels, HbA1c, and histopathology of the liver, kidneys, and pancreas were then assessed.RESULTS: The particle size of the synthesized material was 92.3 nm, which confirmed the nature of nanoparticle. The characteristics of the granule nanoparticle were also in accordance with the standards for drugs suitable for consumption. The administration of GNSP in dose dependent manner significantly decrease blood glucose levels and HbA1C to normal levels compared to control group (p<0.05). Histopathological analysis indicated recovery in pancreas, liver, and kidney tissues following GNSP administration.CONCLUSION: GNSP administration lowers blood glucose levels and HbA1C, as well as improved histopathological condition of pancreas, liver, and kidney in diabetic rats. These findings suggest the potential of utilizing GNSP as a potent antidiabetic agent.KEYWORDS: Citrus Sinensis (L.) Osbeck, histopathology, hyperglycemia, nano medicine, and type 2 diabetes mellitu

Expression of CD8+ and Foxp3+ T Lymphocyte as Predictor for Response to Neoadjuvant Chemotherapy in Stage III Breast Cancer

BACKGROUND: Neoadjuvant chemotherapy (NAC) in breast cancer is usually utilized to eradicate micro-metastasis, induce apoptosis in tumor cells, and reduce the primary tumor size, enabling surgical intervention. Recent studies have shown that tumor-infiltrating lymphocytes (TILs), especially cytotoxic CD8+ T cells and immunosuppressive Foxp3+ regulatory T cells, influence tumor response to treatment. However, their role as predictive markers for NAC response remains unclear. Therefore, this study was performed to investigate whether high expression of CD8+ and low expression of Foxp3+ T lymphocytes are associated with better response to NAC in stage III breast cancer patients.METHODS: Total of 60 biopsy samples from stage III breast cancer patients were included, comprising 30 subjects in the response group (+) and 30 subjects in the non-response group (−). The expression levels of CD8+ and Foxp3+ T lymphocytes in tumor tissue were assessed semi-quantitatively by immunohistochemistry (IHC), using a scoring system based on the proportion and intensity of positively stained cells (Black’s grading criteria).RESULTS: Stage III breast cancer with high expression of CD8+ T lymphocytes was significantly associated with a better response to NAC (p=0.004; OR=6.882). Meanwhile, low expression of Foxp3+ T lymphocytes was not significantly associated with chemotherapy response (p=0.067; OR=3.250). A higher tumor grade was also associated with an improved response to treatment. The probability of achieving a positive response to NAC in subjects presenting with high CD8+ expression, low Foxp3+ expression, and high tumor grade was estimated at 96.98%.CONCLUSION: The combination of high expression of CD8+ T lymphocyte, low expression of Foxp3+ T lymphocyte and high tumor grade might be useful to predict good response to NAC in stage III breast cancer.KEYWORDS: CD8+ T lymphocyte, Foxp3+ T lymphocyte, neoadjuvant chemotherapy, breast cance

Lower Ferrum, Selenium, and Cadmium; Higher Chromium and Lead Levels in Preeclampsia Patients’ Erythrocyte: A Cross-Sectional Study

BACKGROUND: Oxidative stress and trace elements in erythrocytes are linked to impaired nitric oxide that can lead to endothelial dysfunction in preeclampsia patients. The morphology of erythrocytes could also be affected by oxidative stress and trace elements. While the relationships between erythrocyte index, superoxide dismutase (SOD) activity, and oxidative stress in preeclampsia have been well established, less attention has been given to the erythrocyte trace elements and their role in disease progression. This study was performed to examine the erythrocyte trace element profile in women with preeclampsia, comparing it with controls. Additionally, it will explore the correlations between erythrocyte trace element levels, the erythrocyte index, and SOD activity.METHODS: A cross-sectional study was conducted involving 40 pregnant women consisting of those with severe preeclampsia and normotensive. Erythrocytes was isolated from blood samples, and analysis of erythrocyte SOD activity and trace elements were performed using the enzyme linked immunosorbent assay (ELISA) and inductively coupled plasma mass spectrometry (ICP-MS), respectively.RESULTS: Among 15 examined erythrocyte trace elements, the levels of ferrum (Fe), selenium (Se), and cadmium (Cd) were significantly lower, meanwhile, the levels of chromium (Cr) and lead (Pb) were significantly higher in preeclampsia subjects. Additionally, preeclampsia subjects exhibited smaller erythrocyte sizes compared to the normotensive subjects. The erythrocyte SOD activity was significantly elevated in the preeclampsia subjects than the normotensive subjects.CONCLUSION: Erythrocyte trace element levels of Fe, Se, Cd, Cr, and Pb were significantly altered in preeclampsia compared to normotensive controls. These findings suggest that these trace elements may serve as potential predictors for preeclampsia.KEYWORDS: preeclampsia, trace elements, antioxidant, oxidative stress, superoxide dismutase, erythrocyte inde

Roles of Mesenchymal Stem Cell-derived Extracellular Vesicles in Cancer: Development and Target Therapy

Extracellular vesicles (EVs) are membrane structures that enclose proteins, lipids, RNAs, metabolites, growth factors, and cytokines. EVs derived from mesenchymal stem cells (MSCs) can either stimulate or inhibit tumor growth in various malignancies through paracrine signaling. Tumor-associated MSCs (TA-MSCs), often described as "wounds that never heal," actively participate in the development, propagation, and metastasis of tumors, impacting the immunological state of the tumor microenvironment. For instance, TA-MSCs can alter immune cell recruitment and cytokine production, leading to a pro-tumorigenic environment. Consequently, both the tumor and its microenvironment undergo functional alterations, the cargo of exosomes is modified, and an abnormal tumor-associated MSC phenotype is acquired. MSC-EVs contain exosome microRNA with both tumor-inhibitory and tumor-supportive effects. For example, MSC-EVs have been shown to deliver tumor-suppressive microRNAs that inhibit cancer cell proliferation and induce apoptosis. This review outlines the criteria for the modification, isolation, and characterization of exosomes, as well as their application in cancer, providing insights for clinical use. By understanding these mechanisms, we can better harness MSC-EVs for therapeutic purposes.Keywords: mesenchymal stem cell, extracellular vesicle, exosome, cancer therapy, drug deliver