Molecular and Cellular Biomedical Sciences (E-Journal)
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Low Total Lymphocyte Count as the Risk of Hospital Acquired Malnutrition in Children
Background: Hospital Acquired Malnutrition (HAM) is characterized by inadequate nutritional therapy and the risk of developing malnutrition during the hospital stay. In clinical practice, there are many measurements to determine nutritional status. Total lymphocyte count (TLC) is associated with impaired function of immune system in malnutrition. The purpose of this study was to evaluate the prognostic value of TLC to the occurrence of HAM in pediatric patients.Materials and Methods: This an observational study with a prospective cohort design. Subjects were assessed for weight at the first day of hospitalization, then the subjects were followed until they were discharged. Body weight was re-measured on discharge to determine the presence or absence of HAM. This research was conducted at Sanglah Hospital from May-December 2019. Subjects who met the inclusion and exclusion criteria were enrolled in the study.Results: Among 120 subjects, 55 subjects or 45.8% were malnourished on admission. Subjects with a low TLC compared to a normal TLC had a 3.9-fold risk of experiencing hospital acquired malnutrition (95% Confidence Interval: 1.59 to 7.19, p=0.001). Subjects who had a low TLC had HAM of 61.8%, while subjects who had a normal TLC had HAM of 32.3%. In multivariate analysis, low TLC was the only risk factor for HAM in this research.Conclusion: This study proved that low TLC is the risk of HAM. Total lymphocyte count could be used as predictor of the risk of HAM in hospitalization children.Keywords: hospital malnutrition, total lymphocyte, childre
Serum Leptin Concentration is Correlated to Insulin Resistance in Polycystic Ovary Syndrome (PCOS) Patients
Background: Leptin resistance which leads to excessive circulating leptin levels is thought to affect ovarian function. This study aimed to study the correlation between serum leptin levels with insulin resistance in patients with polycystic ovary syndrome.Materials and Methods: This cross-sectional study was undertaken in several teaching hospitals in Makassar, Indonesia. We included patients diagnosed with polycystic ovary syndrome (PCOS) aged 18-40 years old. Serum leptin levels were examined in all eligible subjects using the enzyme-linked immunosorbent assay (ELISA) method. The results obtained were further analyzed statistically.Results: Approximately 53 PCOS subjects were included in this study, 25 subjects with insulin resistance and 28 subjects without insulin resistance. After examining serum leptin levels, we found that leptin is directly proportional to insulin resistance (p<0.001). We even found a strong positive correlation between serum leptin levels with homeostatic model assessment for insulin resistance (HOMA-IR) levels (r=0.659; p<0.001). Leptin was found to be independent of HOMA-IR, not influenced by confounding factors such as body mass index (BMI) (p=0.090).Conclusion: There was a significant correlation between serum leptin levels and HOMA-IR values in PCOS patients. This correlation was found to be significant regardless of patient's BMI, therefore is considered to have a direct effect on insulin resistance in PCOS.Keywords: polycystic ovary syndrome, leptin, insulin resistance, HOMA-I
The Stem Infusate and Ethanol Extract of Physalis angulata Inhibitory Activities against a-Glucosidase and Xanthine Oxidase
Background: Infusate of the whole plant of Physalis angulata is used traditionally for the remedy of various diseases including diabetes and gout. This study focused on the stem of P. angulata. The objectives of this study were to investigate the potential of the stem infusate (INPA) and ethanol extract (EEPA) of P. angulata as inhibitors of α-glucosidase and xanthine oxidase.Materials and Methods: INPA and EEPA were determined for their α-glucosidase and xanthine oxidase inhibition activities in vitro, whereas antioxidant activity was determined by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay. Reference inhibitors were used for comparison. The total phenolic compounds were also estimated.Results: EEPA had more concentrated phenolic than INPA which were 7.96 and 0.08 mgGAE/g dried biomass, respectively. INPA and EEPA inhibited α-glucosidase considerably, with IC50 of 149.11 and 409.86 µg/mL, respectively (acarbose was 130.66 µg/mL). However, they inhibited xanthine oxidase weakly, with IC50 of 0.546 and 2.643 mg/mL, respectively, compared with allopurinol 0.005 mg/mL. EEPA scavenged DPPH radicals very weakly (16.04 mg/mL) compared to BHT (0.021 mg/mL), whereas no activity was observed for INPA.Conclusion: The stem infusate and ethanol extract of P. angulata are able to inhibit the activity of α-glucosidase, thus can be further explored for sources of bioactive compounds with α-glucosidase inhibition activity.Keywords: α-glucosidase, infusate, ethanol extract, Physalis angulata, stem, xanthine oxidas
The Effect of Carnitine Supplementation on Blood Ammonia Level in Epilepsy Children Treated with Valproic Acid: A Randomized Controlled Trial
Background: Long-term use of valproic acid is associated with a high level of blood ammonia related to carnitine deficiency. This study investigates the effect of carnitine supplementation on blood ammonia levels in children with epilepsy who have been treated with valproic acid for more than six months.Materials and Methods: This was a randomized, double-blind, placebo-controlled trial study where children with epilepsy who were treated with valproic acid were randomly allocated to the carnitine supplementation and control group. All children were followed for month, and then measured for blood ammonia level. Blood ammonia levels of both groups were compared using an Independent t-test with a significant of p<0.05.Results: Total of 32 children with epilepsy were enrolled as subjects in this study, with 16 children in carnitine group, and 16 children in control group. Among the subjects, 50% were male and 50% were female, with a mean age of 6.5 years old. The average duration of epilepsy in the carnitine and control group were 41.7 months and 36.9 months, respectively (p=0.419). The duration of valproic acid therapy in the carnitine and control group were 33.1 months and 27.6 months, respectively (p=0.483). The level of blood ammonia in carnitine and control group were 44.6 mg/dL and 81.4 mg/dL, respectively (p=0.007).Conclusion: The level of blood ammonia in a carnitine group was significantly lower than in a control group. It is recommended to give carnitine supplementation in epileptic children treating with long-term valproic acid.Keywords: ammonia, carnitine, epilepsy, seizure, valproic aci
Choline-deficient High-fat Diet-induced Steatohepatitis in BALB/c Mice
Background: Non-alcoholic steatohepatitis (NASH) is an expanding cause of chronic liver disease worldwide, including Indonesia, with higher risk progression to cirrhosis and hepatocellular carcinoma. Preclinical experiments using several mice models have been conducted to clarify its complex pathogenesis. This study was designed to investigate whether BALB/c mice on a choline-deficient high-fat diet can be used as a model for NASH. Materials and Methods: BALB/c male mice were fed choline-deficient L-amino acid-defined high-fat diet (CDAHFD) or a standard diet for six weeks. The body and liver weights, liver histology, and plasma biochemistry were analyzed. The relative expression levels of tumor necrosis factor (TNF)α, transforming growth factor (TGF)β1, collagen-1α1 (COL1α1), glutathione peroxidase 1 (GPx1), and uncoupling protein 2 (UCP2) genes in the livers were analyzed using a two-step real time-polymerase chain reaction. Liver fatty acids composition was analyzed using gas chromatography with flame ionization detector (GC-FID). Results: CDAHFD induced steatohepatitis in BALB/c mice with increased plasma levels of alanine aminotransferase. The liver of CDAHFD-fed BALB/c mice showed upregulated relative expression levels of TNFα, TGFβ1, COL1α1, GPx1, and UCP2 genes. The liver fatty acid analysis showed a significant accumulation of saturated fatty acids (SFAs) and an increased ratio of n-6/n-3 polyunsaturated fatty acids (PUFAs) in the livers of CDAHFD-fed BALB/c mice. Conclusion: This study suggests that CDAHFD can induce steatohepatitis in BALB/c mice and therefore may be used as NASH mice model.Keywords: steatohepatitis, fatty liver, choline-deficient high fat diet, BALB/c
Elevated Serum Reactive Oxygen Species Level Predicts Early Abortion
Background: Impaired trophoblast invasion is associated with early abortion. The calorie needed for the trophoblast cell (TC) invasion is mainly met by adenosine triphosphate (ATP) produced in the mitochondria. Reactive oxygen species (ROS), byproduct of ATP synthesis, plays an important role in cellular physiology, but a high level of ROS may result in deoxyribonucleic acid (DNA) damage or cell dysfunction, thereby impaired TC invasion leading to early abortion. The study aims to determine elevated serum ROS level to predicts early abortion.Materials and method: This was an observational study with a cross-sectional design. Fifty subjects with gestational age less than 12 weeks, consist of 25 early abortions and 25 normal pregnancies subjects, were included in this study. Clinical examination and diagnosis are carried out in 2 Hospitals and 5 Public Health Centers in Padang. Examination of ROS levels was carried out by enzyme-linked immunosorbent assay (ELISA) in the Biomedical Laboratory, Faculty of Medicine, Universitas Andalas. The Mann-Whitney test was used to analyze the difference of serum ROS levels, with a significance level of 0.05.Results: The subjects of the two study groups were equivalent in terms of age, gestational age, and gravidity (p=0.051, p=0.453, and p=1.000). The median ROS levels were found to be 1.36 (1.02-26.30) ng/mL in the early abortion and 1.20 (0.43-2.75) ng/mL in the normal pregnancy (p=0.003).Conclusion: There is a significant difference between ROS levels in early abortion and normal pregnancy.Keywords: ROS, early abortion, normal pregnanc
Effect of Lactobacillus reuteri Administration on Wrinkle Formation and Type I Procollagen Levels in UVB-Exposed Male Balb/c Mice (Mus musculus)
Background: Chronic Ultraviolet B (UVB) exposure causes oxidative stress that may induce damages to the collagen matrix and thus plays a role in the wrinkle formation. Lactobacillus reuteri is a probiotic that may exerts antioxidant effects, thus helping to reduce damages caused by UVB-induced oxidative stress in the skin.Materials and Methods: Twenty-eight male Balb/c mice were divided equally into control group, UVB radiation only group, oral L. reuteri supplementation only group, and UVB radiation with oral L. reuteri supplementation group. UVB irradiation was given 3 times a week (100 seconds/exposure, within 3 cm distance) for 10 weeks, with a total dose of 166 mJ/cm2. Oral L. reuteri supplementation (0.2 mL, 108 CFU) was provided every morning after meal via orogastric feeding tube for 10 weeks. Wrinkle formation on the dorsal skin of the mice was evaluated in accordance with the Bissett method and type I procollagen levels was evaluated by western blotting.Results: In comparison with the group receiving only UVB irradiation, the group receiving probiotic and UVB irradiation showed significantly lower wrinkle scores (Group 1 vs. Group 3, 2.50±0.55 vs. 1.00±0,00; p<0.05) and significantly higher type I procollagen levels (Group 1 vs. Group 3, 0.88±0.36 vs. 1.92±0.46; p<0.05).Conclusion: Results of the current study showed that L. reuteri supplementation may reduce wrinkle formation and increase type I procollagen production in UVB-exposed dorsal skin of male Balb/c mice.Keywords: Lactobacillus reuteri, type I procollagen, photoaging, wrinkles, ultraviolet
Profile of PD-1 and PD-L1 mRNA Expression in Peripheral Blood of Nasopharyngeal Carcinoma
Background: Programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1) expression is associated with prognostic and respond to immunotherapy with immune checkpoint inhibitor in several solid malignancies. However, the prognostic roles of PD-1 and PD-L1 expression in nasopharyngeal carcinoma (NPC) are less clear. This study aims to investigate PD-1 and PD-L1 mRNA expression levels in peripheral blood of Indonesian NPC patients and its association with clinicopathological features.Materials and Methods: This study used blood samples of 21 NPC patients and 10 healthy volunteers as controls. Real-time polymerase chain reaction (PCR) was used to measure mRNA expression of PD-1 and PD-L1.Results: PD-1 mRNA expression levels were significantly lower in NPC patients (∆CT mean: 9.65±2.04) compared to healthy individuals (∆CT mean: 8.04±1.51) (p=0.031). In contrast, PD-L1 mRNA expression levels were higher in NPC patients (∆CT mean: 6.96±1.32) compared to healthy individuals (∆CT mean: 7.11±0.55), but the difference was not statistically significant (p=0.554). The expression of PD-1 was associated with tumour-node-metastasis (TNM) stage (p=0.030) but not associated with age (p=1.000), sex (p=1.000), body mass index (p=0.350), tumor stage (p=0.338), nodal stage (p=0.579), metastasis stage (p=0.371), and Eastern Cooperative Oncology Group (ECOG) status (p=0.228). Meanwhile PD-L1 expression was not associated with all clinicophatological features.Conclusion: The PD-1 mRNA expression levels were significantly lower, while PD-L1 expression levels were higher in NPC patients compared to healthy controls. PD-1 expression was correlated with TNM stage.Keywords: nasopharyngeal carcinoma, immune checkpoint inhibitors, PD-1, PD-L
Antioxidant Activities of Ficus elastica Leaves Ethanol Extract and Its Compounds
Background: The excessive free radicals condition called oxidative stress can harmful for the body. To prevent and cure it, the antioxidant agents are required. Nowadays, the natural product extracted from plants have been widely used in folk medicine as antioxidant for the treatment of many diseases. Ficus elastica (rubber tree) has some compounds that has several biological activities, i.e., quercitrin, myricitrin, morin, and eleutheroside B. The F. elastica works against the free radicals and can be potential as antioxidant agent. The purpose of this study was to evaluate antioxidant properties of F. elastica ethanolic extract (FEE), quercitrin, myricitrin, morin, and eleutheroside B.Materials and Methods: The antioxidant activities of FEE and standard compounds were evaluated by free radical-scavenging activity of 2,2-diphenyl-1-picrylhydrazil (DPPH), hydrogen peroxide (H2O2), 2,2’-azinobis-(3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) activities using spectrophotometry method.Results: FEE has the lowest of DPPH scavenging activity (IC50=13.82 µg/mL) than other compounds. In ABTS scavenging activity, FEE has moderate activity with IC50 value 23.29 µg/mL. In FRAP activity, FEE has moderate activity with value 241.58 µM Fe(II)/µg, while in H2O2 scavenging activity, FEE also show moderate activity with IC50=83.97 µg/mL compared to other compounds.Conclusion: In summary, FEE and the pure compounds (quercitrin, myricitrin, morin, and eleutheroside B) have potential as antioxidant agent.Keywords: free radical, morin, myricitrin, quercitrin, rubber tree, scavenging activitie
Bioassay of Recombinant Human Granulocyte Colony Stimulating Factor (rhG-CSF) for Neutropenia Treatment in Male Sprague Dawley Rats
Background: Recombinant human granulocyte colony stimulating factor (rhG-CSF) is a first line therapy for neutropenia. However, it is less affordable for most patients in developing and poor countries. Therefore, biosimilar products are developed to suppress the cost of treatment, namely with rhG-CSF. This study aimed to explore the establishment of an affordable rhG-CSF that has similar potential to induce neutrophils recovery as the positive control.Materials and Methods: The rhG-CSF was expressed as inclusion body in Escherichia coli NiCo21(DE3). The inclusion body was then solubilized, refolded, purified and characterized prior to be used in the bioactivity assay. Cyclophosphamide-induced male Sprague Dawley rats were used as animal model and administered with rhG-CSF. Blood sample was collected at several points of time, before and after rhG-CSF treatments. Complete blood count and peripheral blood smear were conducted to investigate the activity of the rhG-CSF on each blood cells type, particularly neutrophil.Results: Specific activity on neutrophil proliferation was shown after treatments with our rhG-CSF and positive control. Positive control dose 40 mg/kg BW was statistically similar with that of the rhG-CSF dose 80 and 120 mg/kg BW. However, in neutropenic condition, recovery of neutrophil counts could not be achieved within 4 days of treatments. Thus, a longer treatment is needed to observe the activity of the rhG-CSF as an antineutropenia agent.Conclusion: The rhG-CSF has been proven having specific activity on neutrophil proliferation. However, improvement in the rhG-CSF preparation is still needed and longer administration of the rhG-CSF has to be applied in the future study.Keywords: rhG-CSF, biosimilar, neutropenia, Sprague Dawley rat