Chemical Composition and Neuroprotective Properties of Indonesian Stingless Bee (Geniotrigona thoracica) Propolis Extract in an In-Vivo Model of Intracerebral Hemorrhage (ICH).

Stroke is the world's second-leading cause of death. Current treatments for cerebral edema following intracerebral hemorrhage (ICH) mainly involve hyperosmolar fluids, but this approach is often inadequate. Propolis, known for its various beneficial properties, especially antioxidant and anti-inflammatory properties, could potentially act as an adjunctive therapy and help alleviate stroke-associated injuries. The chemical composition of Geniotrigona thoracica propolis extract was analyzed by GC-MS after derivatization for its total phenolic and total flavonoid content. The total phenolic content and total flavonoid content of the propolis extract were 1037.31 ± 24.10 µg GAE/mL and 374.02 ± 3.36 µg QE/mL, respectively. By GC-MS analysis, its major constituents were found to be triterpenoids (22.4% of TIC). Minor compounds, such as phenolic lipids (6.7% of TIC, GC-MS) and diterpenic acids (2.3% of TIC, GC-MS), were also found. Ninety-six Sprague Dawley rats were divided into six groups; namely, the control group, the ICH group, and four ICH groups that received the following therapies: mannitol, propolis extract (daily oral propolis administration after the ICH induction), propolis-M (propolis and mannitol), and propolis-B+A (daily oral propolis administration 7 days prior to and 72 h after the ICH induction). Neurocognitive functions of the rats were analyzed using the rotarod challenge and Morris water maze. In addition, the expression of NF-κB, SUR1-TRPM4, MMP-9, and Aquaporin-4 was analyzed using immunohistochemical methods. A TUNEL assay was used to assess the percentage of apoptotic cells. Mannitol significantly improved cognitive-motor functions in the ICH group, evidenced by improved rotarod and Morris water maze completion times, and lowered SUR-1 and Aquaporin-4 levels. It also significantly decreased cerebral edema by day 3. Similarly, propolis treatments (propolis-A and propolis-B+A) showed comparable improvements in these tests and reduced edema. Moreover, combining propolis with mannitol (propolis-M) further enhanced these effects, particularly in reducing edema and the Virchow-Robin space. These findings highlight the potential of propolis from the Indonesian stingless bee, Geniotrigona thoracica, from the Central Tapanuli region as a neuroprotective, adjunctive therapy.

Obstacles in Basic Health Service When Dealing with COVID-19: A Reflection for Improvement.

Background & Aims: COVID-19 has been in control since mass vaccination and other coordinated efforts have been conducted. The WHO has even made an announcement to stop the public health emergency of international concern (PHEIC) toward COVID-19. Because of the very massive impacts of COVID-19, it is necessary to have a reflection toward what we have gone through to obtain information from inside the health service by interviewing the health officers who were the frontliners in fighting the disease during the pandemic. This research attempts to reveal the practical experiences of the officers when the COVID-19 pandemic took place. Methods: This research conducted in-depth interviews with the Heads of Puskesmas (a Public Health Center) and the staff of Puskesmas in Medan. The total sample number of informants is 30 people. The data were managed by preparing the suitable themes with the inductive approach. Results: There are three main themes obtained from the analysis results. Those three themes are 1) the condition of Puskesmas, 2) the impacts of the health service, and 3) the COVID-19 control. Those three themes show the fundamental problems affecting the efforts to overcome COVID-19. It is revealed that funding, personnel quantity, and bureaucracy cause the effort to overcome COVID-19 to have been impeded. Meanwhile, based on external factors, the officers had to deal with negative issues on COVID-19. Therefore, whether we like it or not to admit the fact, the health service has not been conducted maximally. Conclusion: It is necessary to have comprehensive revision and change to evaluate the condition of health service, particularly in Puskesmas. A wider and deeper reflection is required so that the pandemic preparation in the future can be improved further.

Study of the Molecular Dynamics Stability in the Inhibitory Interaction of Tenofovir Disoproxil Fumarate against CTLA-4 in Chronic Hepatitis B Patients.

Background: Tenofovir disoproxil fumarate (TDF) is a first-line nucleotide analog (NA) drug for hepatitis B therapy. Long-term NA therapy increases peripheral T cell levels to enhance antiviral response, while CTLA-4 inhibits the activation. Objective: This study analyzed the interaction between TDF and CTLA-4 through molecular docking. Methods: Target protein and ligand data mining were performed, and proteins were prepared by removing water molecules in the Discovery Studio 2019 software. The energy minimization was performed on ligands using Pyrx v.0.9.8 software. Protein-ligand docking was performed using Autodock Vina integrated with Pyrx v.09.8. Meanwhile, the docking of proteins was accomplished using the Haddock server. The BioVia Discovery Studio 2019 software visualized the interaction between the compound and the docked protein. Molecular dynamics simulations were carried out using the YASARA Dynamic program developed by Biosciences GmbH. Results: TDF ligand has good and stable inhibitory activity against the CTLA-4/B7-1 and CTLA4/B7-2 complexes. TDF docking has been shown to initiate conformational changes, indicating the ligand's inhibitory activity. The significant conformational changes based on superimposition results were shown by the CTLA-4/TDF/B7-2 and CTLA-4/B7-1/TDF complexes. TDF in all ligands undergoes bonding and displacement of binding sites. Conclusion: Treatment with TDF was predicted to have inhibitory activity against CTLA-4, especially in its complex form with B7-1 and B7-2.

Phytochemical Analysis by FTIR of Zanthoxylum Acanthopodium, DC Fruit Ethanol Extract, N-hexan, Ethyl Acetate and Water Fraction.

Background: Andaliman fruit is used as spice in Batak cuisine, North Sumatera, Indonesia. The potency of andaliman fruit extract as herbal medicine is widely studied. Many studies elaborate the benefits of andaliman fruit extract as an antioxidant, antibacterial or anticancer. Objective: The aim of this study was to identify the phytochemical compounds of andaliman fruit extract and its fractions. Methods: The andaliman fruit was originated from Balige city, Tobasa Regency, North Sumatera. The extract was made by maceration within ethanol and followed by fractionation with n-hexan, ethyl acetate and water. The phytochemical screening by chemical reactions, thin layer chromatography, and Fourier Transform Infrared Spectrophotometer were performed. Results: This study found andaliman fruit ethanol extract consists of alkaloids, flavonoids, glycosides, tannins, triterpens, and steroids. The n-hexan fraction consists of triterpens and steroids, ethyl acetate fraction consists of flavonoids and glycosides, and water fraction consists of alkaloid, flavonoid, and glycosides. Andaliman fruit ethanol extract had eight color spots, n-hexan and ethyl acetate fraction had five color spots and water fraction had two color spots. Andaliman fruit ethanol extract had functional group of C-H alkanes, =C-H alkenes (aliphatic), O-H, C=C alkenes, C=C aromatics, C≡C alkynes, C-O, C=O, and NO2. The n-hexan fraction had C-H alkanes, =C-H alkenes (aliphatic), O-H, C=C alkenes, C-O, C=O, and NO2. The ethyl acetate fraction had C-H alkanes, =C-H alkenes (aliphatic), O-H, C=C alkenes, C=C aromatics, C-O, C=O, and NO2. The water fraction had C-H alkanes, =C-H alkenes (aliphatic), O-H, C=C aromatics, C≡C alkynes, and C-O. Conclusion: Phytochemical screening found that andaliman fruit ethanol extract, n-hexan, ethyl acetate and water fraction positive of phytochemical compounds. The FTIR of andaliman fruit ethanol extract, n-hexan fraction, ethyl acetate fraction, and water fraction showed the absorbance of C-H alkanes, O-H, C-O, and C=C alkenes indicating the presence of alkaloids, flavonoids, and triterpenoid saponin.

Association Between Serum Cytotoxic T Lymphocyte Antigen (CTLA)-4 Level and Disease Progression in Patients With Chronic Hepatitis B.

Backgroud: Immune impairment, marked by increased expression of cytotoxic T lymphocyte antigen (CTLA)-4, promotes the disease progression of chronic hepatitis B. Objective: This study aimed to determine the association between serum CTLA-4 level and disease progression in patients with chronic hepatitis B. Methods: A cross-sectional study was conducted at Haji Adam Malik General Hospital Medan, Indonesia between October 2021 to September 2022. A total of 150 participants were enrolled. Patients aged 18 years or older with evidence of chronic hepatitis B, HBV-related liver cirrhosis, and HBV-related hepatocellular carcinoma (HCC) were enrolled. Exclusion criteria were history of chronic hepatotoxic drug consumption, underlying liver abnormalities other than HBV infection, and liver injury due to metastasized malignancy from other sites. Serum CTLA-4 level was determined from serum using human CTLA-4 enzyme linked immunosorbent assay kit. Results: Most participants were males and aged between 40 and 60 years. Serum CTLA-4 level was positively associated with chronic hepatitis B progression (P<0.001). Serum CTLA-4 level was negatively correlated with serum platelet (P<0.001) and albumin levels (P<0.001) but positively correlated with serum ALT (P=0.045) and total bilirubin levels (P<0.001). Conclusions: Serum CTLA-4 level is associated with disease progression in patients with chronic hepatitis B.

Degree of kidney injury due to artificial pelvic-ureteric junction obstruction with level of neutrophil gelatinase-associated lipocalin, interleukin 18, and histopathological descriptions in Wistar: experimental research.

Pelvic-ureteric junction obstruction (PUJO) is considered the most common pathology for hydronephrosis in neonates. Full recovery of kidney impairment due to PUJO is possible, especially when pyeloplasty is indicated as surgery is mostly conducted when deterioration is identified, early detection should be considered to prevent further complications. Commonly used kidney damage biomarkers are not sensitive enough to predict kidney damage. Neutrophil gelatinase-associated lipocalin (NGAL) and urinary interleukin 18 (IL-18) are markers of early kidney damage with different characteristics. This study aimed to evaluate the relationship between these two markers with the degree of histopathological kidney damage in Wistar rats induced by PUJO. Methods: A total of thirty male Wistar rats, 200-250 g, were divided into three groups: (1) control, (2) sham, (3) PUJO (4th, 7th, 14th, and 21st days). Urine NGAL, IL-18 levels, and renal histopathology were observed on day 0, 4, 7, 14, and 21. Statistical analysis was performed using the Kruskal-Wallis and Mann-Whitney test with P less than 0.05 considered significant. Results: There was no significant difference in urine NGAL levels between groups, while IL-18 levels were significantly different based on the Kruskal-Wallis test (P 0.031). The results of the Mann-Whitney test showed a significant difference in IL-18 levels between the control group and the PUJO group on day 4 (P=0.028); the Sham surgery group with the PUJO group on day 4 (P=0.014); the PUJO group on day 4 with the PUJO group on the 7th day (P=0.008); and the PUJO group on the 7th day with the PUJO group on the 14th day (P=0.033). Conclusion: Urinary IL-18 levels can be used as a predictor of kidney damage in acute-subacute PUJO cases.

Comparison of Gambier Extract (Uncaria Gambier Robx) and Angiotensin Receptor Blocker on Proteinuria Reduction and Antioxidants - Enhancement in Nephrotic Rat Models.

Background: Proteinuria is a significant clinical manifestation that causes edema in several diseases, including Nephrotic Syndrome (NS). Untreated proteinuria is strongly linked to the progression of kidney failure. One of the adjuvant therapies could be used to reduce proteinuria such as Angiotensin Receptor Blocker (ARB) including Losartan®. Gambier is a traditional medicinal plant widely known for its antioxidant effects. Catechin, a compound contained in Gambier Extract (GE), has been used to reduce microalbuminuria in diabetics. However, its application in NS has not been widely studied. Objective: This study compared the effects of GE and ARB in reducing proteinuria and increasing antioxidant activity levels, as well as reported histopathological findings in the nephrotic Wistar rat model. Methods: An experimental design study with a control group and a posttest was conducted. The experimental animals were divided into four groups: the control group (K1), the group with puromycin aminonucleoside (PAN) injection (K2), the group with PAN injection + GE (K3), and the group with PAN injection + Losartan® (K4). The standard GE used was Sarie Uncariae® by Toyo Brothers, PT while the ARB (Losartan®) was obtained from Novell, PT. Protein urine, the activity level of total superoxide dismutase (T-SOD), and malondialdehyde (MDA) were assessed using the colorimetric method. Renal histopathology was assessed based on Rollerman's criteria. Results: Gambier extract significantly reduced proteinuria, as depicted by a decrease in protein/volume urine (p = 0.009), increased antioxidant activity, as illustrated by an elevation in T-SOD activity levels (p = 0.007), and tended to decrease MDA levels compared to Losartan®. Based on histopathological findings, GE tended to reduce the percentage of kidney damage in rats induced by puromycin. Conclusion: Gambier extract has been shown a higher antioxidant effect by increasing T-SOD activity levels, reducing proteinuria and also exhibiting a tendency to diminish kidney damage.

Potential benefit of olive leaf extract in cervical spondylotic myelopathy model.

INTRODUCTION: Cervical spondylosis is the most common cause of myelopathy in the cervical due to chronic compression of the spinal cord in patients aged 55 years or older. Recent studies suggest that olive extracts suppress inflammation and reduce stress oxidative injury. The purpose of this study was to determine the potential neuroprotective effects of olive leaf extract (OLE) in an experimental cervical spondylotic myelopathy model. METHODS: This study was divided into 6 groups; Control Negative (Sham-Operated) Group, Control Positive 1 & 2 (early chronic and chronic), Treatment Groups 1, 2 & 3 (prophylactic, concomitant & late). Olive leaf extract (OLE) give 350 mg/kg BW and spinal cord sample was taken at the compression level C5. Histopathological assessment and immunohistochemistry of Amyloid-ß, p-Tau, TDP-43 dan CD-68 dan evaluation of functional motoric outcome was done before animals were terminated. RESULTS: Chronic spinal cord compression increased the expression of Amyloid-ß, p-Tau, TDP-43 dan CD-68. OLE 350 mg/kg BW decreased the expression of these biomarkers and increased functional motoric outcome, especially as prophylactic dan concomitant treatment. DISCUSSION: These findings indicate that OLE may be effective in protecting cervical spondylotic myelopathy.

Effectiveness of upper limb exercises and breathing exercises in COPD stable patients during COVID-19 pandemic era.

MATERIAL AND METHODS: This study was a quasi-experiment conducted from August to November 2020 involving 10 COPD stable patients. In the first meeting at the hospital, patients were taught breathing techniques and upper-limb exercise movements which subsequently had to be done and recorded at home. Patients then sent the videos to the research team via whatsapp to be evaluated. Prior to starting the 4-week with twice-a-week frequency home-based rehabilitation, and again at the end of it, patients' conditions were evaluated based on the following factors: lung functional capacity was measured by 6MWD (6 Minutes Walking Distance), Perceived Exertion Scale by mMRC (modified Medical Research Council) and Borg, the quality of life by SGRQ (St. George's Respiratory Questionnaire) index, the severity of symptoms by CAT (COPD Assessment Test) and changes in muscle mass. Statistical analysis was conducted using the Wilcoxon and paired t-test. RESULTS: After 4 weeks of home training with twice-a-week frequency, the results showed significant improvement, with p-value < 0.05, in the increased 6MWD value (16.4 ± 4.45 to 19.8 ± 3.64), mMRC (1.90 ± 1.10 to 0.90 ± 0.87), Borg (3.05 ± 1.53 to 1.40 ± 1.42), CAT (15.7 ± 6.63 to 11.8 ± 5.24) and muscle mass (28.36 ± 2.89 to 31.86 ± 3.97). CONCLUSION: The method produced positive outcomes in patients' lung function capacity, severity of symptoms, exertion scale, muscle mass, and general quality of life. Accordingly, the home-based rehabilitation programme was as effective as the programme conducted at health-care centers.

Factors affecting HBV DNA suppression in chronic hepatitis B patients treated with tenofovir disoproxil fumarate.

Background: This study aims to determine the factors affecting HBV DNA suppression in chronic hepatitis B patients with tenofovir disoproxil fumarate (TDF). Methods: A case-control was carried out from October 2021 to August 2022 on 182 chronic hepatitis B patients who had TDF therapy regularly for 24 weeks at H. Adam Malik and USU Hospitals in Medan, Indonesia. The history of the samples was obtained, followed by physical examination, and blood collection. CTLA-4 polymorphism examination was carried out using real-time PCR, while the serum CTLA-4 levels were assessed with ELISA. Results: The CTLA-4 -1661G>A polymorphism, genotype GG+AG, increased 1.52 times risk of not achieving HBV DNA suppression to TDF compared to genotype AA (p=0.041). High CTLA-4 levels increased 2.28 times risk, high HBV DNA levels increased 2.09 times risk, low ALT levels increased 1.95 times risk of not achieving HBV DNA suppression (p= 0.009, 0.026, 0.036, respectively). There was no relationship between gender, age, ethnicity, obesity, baseline AST, HBeAg, genotype, liver fibrosis and HBV DNA suppression after 24 weeks of treatment (p>0.05). Conclusions: The levels of CTLA-4, HBV DNA, ALT, and CTLA-4 -1661G>A polymorphism have a potential relationship with the suppression of HBV DNA in chronic hepatitis B patients with TDF.

Human avian influenza in Indonesia: are they really clustered?

Understanding the epidemiology of human H5N1 cases in Indonesia is important. The question of whether cases are clustered or not is unclear. An increase in clustered cases suggests greater transmissibility. In the present study, 107 confirmed and 302 suspected human H5N1 cases in Indonesia during 2005-2007 were analyzed for spatial and temporal distribution. Most confirmed cases (97.2%) occurred on two main islands (Java and Sumatera). There were no patterns of disease occurrence over time. There were also no correlations between occurrence patterns in humans and poultry. Statistical analysis showed confirmed cases were clustered within an area on Java island covered by 8 districts along the border of three neighboring provinces (Jakarta, West Java, and Banten). This study shows human H5N1 cases in Indonesia were clustered at two sites where there was a high rate of infection among poultry. These findings are important since they highlight areas of high risk for possible human H5N1 infection in Indonesia, thus, preventive measures may be taken.