Daily Soy–Catfish–Anchovy–Rice (SCAR) Porridge Increases 25(OH)D Serum Levels in Tuberculosis Patients with Vitamin D Receptor Gene Polymorphisms

BACKGROUND: The presence of vitamin D receptor (VDR) polymorphism and high levels of inflammatory markers are predisposing factors indicating disease progression and malnutrition. To meet nutritional needs in maintaining nutritional status in tuberculosis patients with VDR gene polymorphisms (TaqI or FokI), food that is easily absorbed and high in vitamin D, calcium, and protein is needed. This study was conducted to determine whether high vitamin D and calcium porridge called soy–catfish–anchovy–rice (SCAR) porridge would increase 25(OH)D serum levels and other parameters, including calcium, albumin, high-sensivity C-reactive protein (hs-CRP), and blood glucose serum levels.METHODS: The study was a parallel, open, clinical trial. There were 22 subjects in the intervention (I) group who received 50 g of SCAR porridge once per day along with dietary counseling, and 21 subjects in the control (C) group who only received dietary counseling. All subjects with TC and CC (TaqI) or TC and CC (FokI) genotype were included in this study. The intervention lasted for 14 days, and the parameters such as 25(OH)D, calcium, albumin, hs-CRP, and blood glucose serum levels were assessed before and after intervention between groups.RESULTS: All subjects in both groups completed the study. After 14 days of intervention, there was a significant increase in 25(OH)D (p=0.01) and decrease in hs-CRP (p=0.02) serum levels in the I group, and no change was observed in the C group. There was no significant difference in albumin and blood glucose serum levels.CONCLUSION: The results show that 50 g of SCAR porridge per day for 14 days is an effective supplementation that can increase 25(OH)D and decrease hs-CRP serum levels in tuberculosis patients with VDR gene polymorphism.KEYWORDS: vitamin D, hs-CRP, polymorphism, porridge

Higher Neutrophil-lymphocyte Ratio in TB/HIV Co-infection Compared to Pulmonary Tuberculosis

BACKGROUND: Neutrophils and lymphocytes play a significant role in inflammation and a high ratio of neutrophils over lymphocytes (NLR) has been used as an inflammatory marker to predict the severity of various diseases. Here we compared the NLR among pulmonary tuberculosis and TB/HIV co-infection.METHODS: A retrospective cross-sectional study was conducted, included patients with pulmonary TB without cavitation TB (n=50), with cavitation TB (n=50) and HIV co-infection (n=27). Complete blood count was examined, including neutrophils and lymphocyte. NLR was calculated and compared between groups. RESULTS: Neutrophils were significantly higher (p=0.004) in TB with cavitation compared to those with no cavitation (8.27±1.45 x103/μL vs. 6.61±1.4 x103/μL, respectively); whereas the lymphocytes were similar in both groups, resulting in a significantly higher NLR (p=0.009) in pulmonary TB with cavitation compared to pulmonary TB with no cavitation (5.98±1.85 vs. 4.42±1.86, respectively). On the contrary, both neutrophils as well as lymphocyte were significantly lower in TB/HIV compared to pulmonary TB, which for neutrophil were 5.14±2.19 x103/μL vs. 7.4±1.45 x103/μL, respectively (p=0.003) and for lymphocyte (1.02 ±0.57 x103/μL vs. 1.57±0.64 x103/μL, respectively (p=0.001), resulting in a significantly higher (p=0.041) NLR value in TB/HIV (6.05±2.67) compared to pulmonary TB (5.16±1.88).CONCLUSION: High NLR in pulmonary TB with cavitation as well as in TB with HIV co-infection may be of great interest for biomarker in TB severity. Further study confirming NLR as potential marker is imperative.KEYWORDS: lymphocyte, neutrophil, NLR, tuberculosis, TB/HIV

Association of 25-hydroxyvitamin D, Cyclooxygenase-2 and Prostaglandin E2 Serum Levels in Breast Cancer Patients

BACKGROUND: Serum levels of 25-hydroxyvitamin D (25(OH)D), prostaglandin E2 (PGE2), and cyclooxygenase 2 (COX2) expression differ between breast cancer stages. Since, previous studies showed mixed results, in this study, we aimed to analyze vitamin D levels related to breast cancer stages and serum levels of COX2 and PGE2 in Indonesia.METHODS: This was a cross sectional study involving 75 breast cancer patients. Subjects were divided into 3 groups, namely operable early stage (K1), locally advanced stage (K2), and advanced stage (K3). Venous blood samples were taken from each subject, then were analyzed for the 25(OH)D, COX2, and PGE2 serum levels by enzyme-linked immunosorbent assay (ELISA) method.RESULTS: There were significant differences in 25(OH)D among groups (p=0.012); between K1 and K2 (p=0.009) and between K1 and K3 (p=0.023). However, there was no significant difference in serum COX2 level (p=0.328). There were significant differences of PGE2 among groups (p=0.002); between K1 and K2 (p=0.036) and between K1 and K3 (p=0.001). Correlation test showed that there were differences between 25(OH)D serum levels and PGE2 serum level (r=0.306, p=0.008) and also between 25(OH)D serum level and breast cancer stage (r=-0.229; p=0.048).CONCLUSION: There were differences in serum Vitamin D and PGE2 levels at various stages of breast cancer. Serum 25(OH)D levels had weak correlation with breast cancer stage and PGE2 serum level. Serum vitamin D level in advanced breast cancer were lower than early stage breast cancer and indicate a poor prognosis.KEYWORDS: breast cancer, 25-hydroxyvitamin D, cyclooxygenase 2, prostaglandin E2

Serumal and Salivary 25(OH)D and 1,25(OH)D Levels of Head and Neck Cancer Patients

BACKGROUND: Saliva has been suggested as a substitute of serum for the detection of 25 Dihydroxyvitamin D (25(OH)D) in healthy people. However, investigation of salivary 1,25(OH)D has not been clearly reported. Vitamin plays important roles in inhibiting cancer progression. Current study was conducted to investigate serumal and salivary 25(OH)D) and 1,25(OH)D levels of healthy and head and neck cancer (HNC) subjects.METHODS: Research were conducted at Haji Adam Malik Hospital, Medan, Indonesia. Forty HNC and 40 healthy subjects were recruited and selected based on inclusion and exclusion criteria. Medical records were documented, followed by anthropometric evaluation and serum and saliva collection. Laboratory investigation for 25(OH)D and 1,25(OH) was performed using Enzyme-linked immunosorbent assay (ELISA) methods.RESULTS: Significant serumal (p=0.002) and salivary (p=0.016) 25(OH)D mean level differences of HNC and normal groups were obtained. More serumal or salivary 25(OH)D deficient subjects were found in control group than those in HNC group. Meanwhile, serumal and salivary 1,25(OH)D mean levels of HNC group were not significantly different with the ones of control group. There were significant correlations of serumal-salivary 25(OH)D as well as serumal-salivary 1,25(OH)D levels in normal group.CONCLUSION: Serumal and salivary 25(OH)D and 1,25(OH)D levels of HNC group were relatively normal. Salivary 25(OH)D and 1,25(OH)D could be suggested as substitutes for serumal ones.KEYWORDS: vitamin D, 25(OH)D, 1,25(OH)D, head and neck cancer

Caffeic Acid Inhibits RANKL and TNFa-induced Osteoclastogenesis by Targeting TAK1-p44/42 MAPK

BACKGROUND: The potential of the caffeic acid in other important Receptor Activator Nuclear Factor kB Ligand (RANKL)-Tumor Necrosis Factor (TNF)a-induced osteoclastogenic signaling pathways has not been known. Therefore, the current study was conducted to explore as well as to understand the inhibition potential of caffeic acid.METHODS: RAW264.7 cells were cultured, treated with caffeic acid, RANKL and TNFa. Tartrate Resistant Acid Phosphatase (TRAP) staining was performed to detect TRAP+ osteoclast-like polynuclear cells. To detect the activity of p44/42 Mitogen Activated Protein Kinase (MAPK), Akt, and Transforming Growth Factor-β-activated Kinase (TAK)1, the phosphorylated forms of the proteins were investigated with the immunoblotting assay.RESULTS: Pre-treatment of caffeic acid inhibited the RANKL and TNFa-induced differentiation of RAW264.7 cells into TRAP+ osteoclast-like polynuclear cells. RANKL and TNFa induced phosphorylation of p44/42 MAPK at Thr202/Tyr204, phosphorylation of Akt at both Ser473 and Thr308 and phosphorylation of TAK1 at Ser412. Pre-treatment with caffeic acid prior to the RANKL and TNFa induction, inhibited the phosphorylation of MAPK, and TAK1, but not Akt.CONCLUSION: Caffeic acid might regulate the RANKL-TNFa-induced osteoclastogenic pathway in RAW264.7 by targeting TAK1, which later activation of p44/42 MAPK was abolished.KEYWORDS: caffeic acid, osteoclastogenesis, p44/42, Erk1/2, Akt, TAK1, RAW264.7

Lower GCS is Related to Poor Outcome among Acute Stroke Patients with COVID-19 in A Tertiary Referral Hospital in Indonesia

BACKGROUND: The Coronavirus disease 2019 (COVID-19) pandemic has forced the health workforce to take mitigative measures such as physical distancing, screening, personal protective equipment donning, and confinement on patient care. We aimed to study the outcome of acute stroke patients with suspected, probable, or confirmed COVID-19 in a tertiary referral hospital in Indonesia during the first year of COVID-19 pandemic.METHODS: This was a retrospective study examining all medical records of adult patients suffering from acute stroke with suspected or confirmed COVID-19 who were admitted to R.D. Kandou Hospital, Manado, Indonesia, between March 2020 to March 2021. Clinical and laboratory parameters were compared between subjects with poor and good outcomes based on Glasgow Outcome Scale (GOS), divided into poor outcome (GOS 1-3) and good outcome (GOS 4-5).RESULTS: Fourty-six eligible subjects were enrolled in the study. Based on the GOS, 36 subjects (78.3%) were admitted to the hospital with poor prognosis. On admission, the median Glasgow Coma Scale (GCS) was 11, breathlessness was found in 54.3% of subjects, fever was found in only 15 subjects (32.6%), and the lowest oxygen saturation on admission 95%. We found that GCS significantly related to outcome after controlled for other factors using the logistic regression method (p=0.03; 95% CI=1.08-4.78).CONCLUSION: Lower GCS can be used to predict poor outcome in acute stroke patients with COVID-19.KEYWORDS: COVID-19, acute stroke, Glasgow Coma Scale, outcome, Indonesia

Antihypercholesterolemic and Antioxidant Effects of Blumea balsamifera L. Leaf Extracts to Maintain Luteinizing Hormone Secretion in Rats Induced by High-Cholesterol Diets

BACKGROUND: The discovery of herbal ingredients for antihypercholesterolemic and to improve male reproductive function is very necessary due to high-cholesterol diet factors. Blumea balsamifera leaf extract (BBLE) is known to be able to increase the number and diameter of Leydig cells in rats given high-fat feed. This study was to conducted to determine the levels of total cholesterol, body weight, plasma malondialdehyde (MDA), Superoxide Dismutase (SOD), Luteinizing Hormone (LH) in high-cholesterol-fed male rats provided with BBLE.METHODS: This research utilized a randomized post-test only control group. Ethanol was used to extract the BBLE, which was then evaporated. For 21 days, 16 Wistar rats were given a high-cholesterol diet. To determine the effect of BBLE on the high-cholesterol diet, the samples were divided into two groups (control and BBLE group) on day 22. The treatments lasted 30 days. SOD, plasma MDA, LH, and total cholesterol were measured.RESULTS: The results showed that the SOD and LH parameters were significantly higher in the treatment of BBLE compared to the control group (p<0.05). The parameters of total cholesterol levels, bodyweight, and MDA of rats given BBLE were significantly lower than those of the control group (p<0.05).CONCLUSION: Our findings highlight that BBLE has antihypercholesterolemic and antioxidant effects. The BBLE also has potential to be used as a therapy to maintain male reproductive function because it has a positive effect on the hypothalamic-pituitary axis through increasing LH secretion.KEYWORDS: Malondialdehyde, Superoxide dismutase, Luteinizing hormone, Blumea balsamifera extract, High-cholesterol diet

Suppression of MiR130a-3p Using CRISPR/Cas9 Induces Proliferation and Migration of Non-Small Cell Lung Cancer Cell Line

BACKGROUND: Molecular alterations of microRNA130a (miR130a) are observed in many types of cancers, including non-small cell lung cancer (NSCLC). However, the role of miR130a in NSCLC has been poorly studied.METHODS: In this study, clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 was utilised to knockdown miR130a. The gRNA was designed to target the stem loop, 3’ and 5’ sites of miR130a and stably expressed in A549 cells. Post-treatment, mature levels of miR130a-3p and 5p were quantified, and proliferation and migration assays were conducted.RESULTS: Result showed significant suppression of miR130a-3p and -5p by two and three-fold respectively, when the CRISPR/Cas9 targeted at the 3’ site and stem loop of the miR130a gene. Suppression of miR130a-3p significantly increased the growth and migration of A549 cells, but no significant changes were observed in cells with suppressed expression of miR130a-5p.CONCLUSION: Our encouraging results highlight that the suppression of the miR130a is achievable using CRISPR/Cas9, and suppression of the miR-130a-3p could play an important role in the regulation of NSCLC.KEYWORDS: miR130a, CRISPR-Cas9, non-small cell lung cancer

A Pilot Study on Immunohistochemical Expressions of NF-ĸB, Cyclin-D1, VEGF, and Cox-2 in Advanced Stage Laryngeal Carcinoma

BACKGROUND: Progression of laryngeal carcinoma can be classified with the clinical staging, however there are different patterns of progressions observed in the patient with the same clinical stage which also affects their prognoses. Therefore biomarkers should be used. Nuclear factor (NF)-ĸB, Cyclin-D1, vascular endothelial growth factor (VEGF), cyclooxygenase (Cox)-2 have been reported for laryngeal carcinoma. However, it is still unclear how these markers are expressed and correlated in advanced stage laryngeal carcinoma. Therefore current study was conducted to investigate the expressions of NF-ĸB, Cyclin-D1, VEGF and Cox-2 and their correlations in advanced stage laryngeal carcinoma.METHODS: Subjects were recruited and laryngeal biopsies were collected, fixed in formalin and prepared for immunohistochemistry. The immunohistochemistry was performed using mouse monoclonal anti-NF-kB p65, anti-Cyclin-D12 anti-VEGF, and anti-Cox-2 antibodies. The immunohistochemistry results were documented and measured using ImmunoRatio. Pearson or Spearman correlation test was used based on the results of Shapiro-Wilk test of normality. A p-value of less than 0.05 is considered statistically significant.RESULTS: Twelve male subjects were included in this study. Expressions of NF-ĸB, Cyclin-D1, VEGF dan Cox-2 were clearly observed. Mean of NF-ĸB, Cyclin-D1, VEGF dan Cox-2 IHC expression levels measured with ImmunoRatio were 57.50±20.06%, 45.00±24.31%, 43.33±17.23% and 40.42±16.98%, respectively. There was significant correlation between the expressions of VEGF dan Cox-2 (p=0.031, r=0.622).CONCLUSION: Since correlation between the VEGF and Cox-2 expressions was statistically significant, VEGF and Cox-2 might have important roles in the growth, invasion and metastasis of laryngeal carcinoma.KEYWORDS: advanced stage laryngeal carcinoma, immunohistochemistry, NF-ĸB, Cyclin-D1, VEGF, Cox-2

Effect of Cell Culture Medium on the Proliferation and Stemness of CD24-/CD44+ Human Breast Cancer Stem Cells

BACKGROUND: Cancer stem cells (CSCs) is defined as tumor initiating cells within tumor that maintain stemness properties and tumorigenicity. Extracellular pH of CSCs in in vitro condition is important for supporting cell proliferation which may also regulate the expression of stemness markers such as OCT4. This work aimed to examine the effect of cell culture media on the proliferation and stemness of human breast cancer stem cells (BCSCs).METHODS: Human CD24-/CD44+ BCSCs were grown in Dulbecco's Modified Eagle Medium/F-12 (DMEM/F-12) with 15mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), without HEPES and adjusted to pH 7.4, or without HEPES but pH was not adjusted. BCSCs were grown under standard conditions for various days. Viable cell number was measured using trypan blue exclusion, whereas proliferation rate using MTS assay. OCT4 mRNA and protein were analyzed using quantitative real time PCR (qRT-PCR) and Western Blot assay, respectively. In vitro tumorigenic activity was determined using mammosphere formation unit (MFU) assay.RESULTS: Our results showed a higher viable cell number and proliferation of BCSCs in DMEM/F-12 HEPES (-) compared to HEPES (+) medium until 4 day incubation. OCT4 mRNA and protein level, as well as MFU of BCSCs were significantly higher in HEPES (-) compared to HEPES (+) medium on day 2.CONCLUSION: DMEM/F-12 medium without HEPES facilitates CD24-/CD44+ BCSCs to have higher proliferation and stemness on day 2 incubation compared to those with HEPES.KEYWORDS: breast cancer, cancer stem cell, OCT4, stemness, proliferation