Analysis of Antidiabetic Activity of Squalene via In Silico and In Vivo Assay.

Squalene has been tested widely in pharmacological activity including anticancer, antiinflammatory, antioxidant, and antidiabetic properties. This study aims to examine antidiabetic activity of squalene in silico and in vivo models. In the in silico model, the PASS server was used to evaluate squalene antidiabetic properties. Meanwhile, the in vivo model was conducted on a Type 2 Diabetes Mellitus (T2DM) with the rats separated into three groups. These include squalene (160 mg/kgbw), metformin (45 mg/kgbw), and diabetic control (DC) (aquades 10 mL/kgbw) administered once daily for 14 days. Fasting Blood Glucose Level (FBGL), Dipeptidyl Peptidase IV (DPPIV), leptin, and Superoxide Dismutase (SOD) activity were measured to analysis antidiabetic and antioxidant activity. Additionally, the pancreas was analysed through histopathology to examine the islet cell. The results showed that in silico analysis supported squalene antidiabetic potential. In vivo experiment demonstrated that squalene decreased FBGL levels to 134.40 ± 16.95 mg/dL. The highest DPPIV level was in diabetic control- (61.26 ± 15.06 ng/mL), while squalene group showed the lowest level (44.09 ± 5.29 ng/mL). Both metformin and squalene groups showed minor pancreatic rupture on histopathology. Leptin levels were significantly higher (p < 0.05) in diabetic control group (15.39 ± 1.77 ng/mL) than both squalene- (13.86 ± 0.47 ng/mL) and metformin-treated groups (9.22 ± 0.84 ng/mL). SOD activity were higher in both squalene- and metformin-treated group, particularly 22.42 ± 0.27 U/mL and 22.81 ± 0.08 U/mL than in diabetic control (21.88 ± 0.97 U/mL). In conclusion, in silico and in vivo experiments provide evidence of squalene antidiabetic and antioxidant properties.

Water Extract Have Superior Cytotoxic Effect Than Ethanolic Extract of Clinacanthus Nutans Leaves in Breast Cancer Stem Cells.

Background: The rapid development of medical technology in managing breast cancer patients still cannot solve the problem of recurrence and resistance. One of the causes of recurrence and molecular resistance is the presence of breast cancer stem cells (BCSCs). Clinacanthus nutans (C.nutans) is a plant found in Medan, North Sumatra, Indonesia. This plant is believed to have anticancer activity in community. Objective: Our study aimed to assess phytochemical of C.nutans leaves, isolate breast cancer stem cells and determine the cytotoxic effects of the ethanolic extract and water extract of C.nutans leaves on breast cancer stem cells at 24, 48, and 72 h of observation. Methods: We underwent the cytotoxic test by using MTT assay and isolated breast cancer stem cells by using MACS and validated them by mammosphere test. Results: We found alkaloids, flavonoids, glycosides and tannins in simplicia and all extracts. BCSCs was valid with the diameter of the mammosphere BCSCs was > 60 µm. The IC50 values of 100%, 60%, 40%, 20% EE, and WE of C.nutans leaves were 227.30; 46.05; 31.12; 98.54, and 16.16 µg/ml respectively in the first 24 hours. In administering WE of C.nutans leaves, BCSCs viability was decreased at 24,48 and 72 hours of observation, namely 69.29±26%; 75.82 ± 21.02% and 38.94±9.34 % (p < 0.0001). Conclusion: The WE of C.nutans leaves had more substantial cytotoxic potential against BCSCs than the EE. The capability of WE C.nutans leaves to suppress BCSC's viability was time-dependent. The anticancer activity were believed originate from alkaloid and flavonoid group.

The Relation of Haplotype ATP-binding Cassette B1 and Glutathione S-transferase P1 A313G Genes with Hematological Toxicity in Indonesian Breast Cancer Patients Receiving Chemotherapy.

Objectives: Hematological toxicity induced by chemotherapy is known to be caused by multiple factors, including genetic factors such as polymorphisms. The polymorphisms may occur in drug efflux transporter proteins and enzymes involved in drug metabolism. We investigate the incidence of hematological toxicities and their relation to the haplotype ATP-binding cassette B1 (ABCB1) which were polymorphisms of C1236T, C3435T, G2677T, and glutathione S-transferase P1 (GSTP1) A313G genes in Indonesian breast cancer patients who received anthracycline during chemotherapy. Methods: This retrospective cohort study was conducted on 138 breast cancer patients who underwent three cycles of chemotherapy in H. Adam Malik Hospital, Medan, Indonesia, who satisfied the inclusion criteria. The DNA of these patients was extracted from the peripheral leukocytes. Single nucleotide polymorphism (SNP) ABCB1 and GSTP1 were examined by the polymerase chain reaction-restriction fragment length polymorphism method. Data on patient characteristics and the incidence of hematological toxicity after each of the three cycles of chemotherapy were obtained from the medical records. The variations in absolute neutrophil count (ANC) and anemia were analyzed using the Friedmann test and the Wilcoxon signed-rank test. The Kruskal-Wallis test was used to investigate the association of ABCB1 and GSTP1 polymorphisms with anemia and neutropenia. The frequency distributions of genotypes and alleles were determined using the Hardy-Weinberg Equilibrium (HWE). Results: Post the chemotherapy cycles, there was decrease in ANC (Mean±SD: 5 644.4±2 962.5 mm3 vs. 3 034.8±2 049.6 mm3) and increase in anemia (12.1±1.5 g/dL vs. 11.2± 1.3 g/dL) (p < 0.050 for each). No relation was observed between ABCB1 polymorphism, either in each SNP or in the form of haplotype (the combination of more than one SNP), and the incidence of anemia and neutropenia (p > 0.050). There was also no correlation between GSTP1 polymorphisms, anemia and neutropenia incidence (p > 0.050). The ABCB1 and GSTP1 genotypes and alleles frequency distribution showed no deviation from HWE (p > 0.050). Conclusions: Indonesia breast cancer patients who underwent three cycles of chemotherapy demonstrated susceptibility to hematological toxicity by developing side effects such as anemia and neutropenia. However, no relationship was found between hematological toxicity and ABCB1 and GSTP1 polymorphisms.

The Gene Polymorphisms (-308G/A) and the Tumor Necrosis Factor-alpha Levels in Type 2 Diabetic Patients with and Without Tuberculosis Infection.

BACKGROUND: The gene polymorphism (-308G/A) and tumor necrosis factor-alpha (TNF-α) levels influence development of disease in type 2 diabetic patients and tuberculosis patients. AIM: In this study, we analyze the association between the TNF-α polymorphisms (-308G/A) and the levels of TNF-α in type 2 diabetic patients with and without tuberculosis infection. METHODS: This study was an analytic observational with cross sectional approach consisting 40 type 2 diabetic patients with tuberculosis infection, 40 type 2 diabetic patients without tuberculosis infection and 40 healthy control (HC) subjects. The TNF-α gene polymorphism (-308G/A) was analyzed with polymerase chain reaction-restriction fragment lengths polymorphisms (PCR-RFLP) method. The TNF-α levels were measured using an enzyme-linked immunosorbent assay. The association between gene polymorphism (-308G/A) in study groups was analyzed by Fisher's exact test, tumor necrosis factor-alpha (TNF-α) levels in study groups was carried out using the Kruskal-Wallis test. Hardy-Weinberg Equilibrium also determined genotype deviation and allele frequencies. RESULTS: The GG and GA+AA genotypes frequency in both of patient groups and HC subjects were not differ significantly (95% and 5% vs 95% and 5% vs 92.5% and 7.5%; p > 0.05). The TNF-α levels (pg/ml) of type 2 diabetic without tuberculosis infection were higher than those of type 2 diabetic with tuberculosis infection and HC subjects (7.42 ± 0.78 vs 2.23 ± 0.51 vs 2.57 ± 0.63; p < 0.01). The TNF-α levels in the GA+AA genotypes were higher than the GG wild-type genotype (p > 0.05). There was no significant deviation of genotype frequency and allele from Hardy-Weinberg Equilibrium. CONCLUSION: The gene polymorphism (-308G/A) had no association with type 2 diabetic patients with and without tuberculosis infection and the gene polymorphism (-308G/A) was not influence the TNF-α levels but there was a significant differentiation of TNF-α levels between the groups.