UC-MSCs appear to be a viable option for stem cell-based treatment. Cynomolgus monkeys, with their psychological and genetic parallels to humans, are essential models for medical study and development. Hypoxia is a basic characteristic of angiogenesis, glucose metabolism, and cell proliferation and survival in both healthy and pathological individuals. This study aimed to develop and characterize UC-MSCs requirements derived from the cynomolgus monkey. We have successfully developed UC-MSCs derived from Mf under hypoxic conditions. Mf UC-MSCs was cultured from the WJ area. Our Mf UC-MSCs have developed into adipocytes, osteocytes, and chondrocytes. The protein of the biomarker MSC was expressed in our hypoxic precondition Mf UC-MSCs and the mRNA expression levels of ERK1/2, AKT1, and HIF-1 alpha increased in the hypoxic condition compared to the normoxic and control groups. The JNK and NFκB genes were decreased in the hypoxic group. Interestingly, our hypoxic Mf UC-MSCs suppressed IL-6 expression. These findings show that preconditioned Mf UC-MSCs produce anti-inflammatory properties, making them potential candidates for regenerative medicine. Their response to hypoxia is crucial for developing targeted therapies to enhance regenerative capabilities in conditions such as ischemic injury and neurodegeneration.

Purpose: Mesenchymal stem cells (MSCs) hold therapeutic promise for ischemic stroke through paracrine release of bioactive factors that promote angiogenesis and neurogenesis. This study evaluated the neurogenic potential of Macaca fascicularis UC-MSC secretome under hypoxic and normoxic conditions in an in vitro ischemic stroke model, and assessed its promise as a regenerative, cell-free therapy. Methods: mUC-MSCs were cultured under normoxic (21% O₂) and hypoxic (3% O₂) conditions for 48 hours. Secretomes were collected and analyzed for Brain-Derived Neurotrophic Factor (BDNF) and Stromal Cell-Derived Factor-1 (SDF-1) levels via enzyme-linked immunosorbent assay (ELISA), while exosome morphology was characterized using scanning electron microscopy (SEM). An in vitro ischemic stroke model was created using SH-SY5Y cells exposed to 6 hours of oxygen-glucose deprivation (OGD) followed by 24 hours of reperfusion. During reperfusion, cells were treated with normoxic or hypoxic secretome. Neurogenesis-related gene expression was assessed by RT-PCR and statistically analyzed. Results: The secretome from 3% O2 hypoxic preconditioning showed elevated levels of BDNF and SDF-1 compared to normoxic conditions. RT-qPCR analysis revealed significant upregulation of neurogenesis-related genes BDNF, TrkB, ERK1/2, Nestin, and β-tubulin following treatment with the hypoxic secretome, indicating enhanced neurogenic activity and neuronal survival in the in vitro ischemic stroke model. Conclusion: Hypoxia preconditioned UC-MSC secretome enhances neurogenesis by upregulating key neurogenic markers, promoting neuronal differentiation and regeneration in ischemic conditions. These results highlight the promise of this approach as a regenerative, cell-free therapy for promoting recovery following ischemic stroke.

The current treatment for ischemic stroke, tissue plasminogen activator (tPA), is limited by a narrow therapeutic window, necessitating the development of adjuvant therapies. Hypoxic preconditioning is a strategy to improve the therapeutic potential of mesenchymal stem cells. The secretome of hypoxic mesenchymal stem cell-conditioned medium (hypMSC-CM) is an effective therapy for vascular lesions like stroke, as it promotes angiogenesis, neurogenesis, and neuroprotection. This systematic review aimed to assess the efficacy of hypMSC-CM in in vivo and in vitro stroke models to inform future clinical trials on stroke treatment. A systematic search was conducted for studies on hypMSC-CM in ischemic stroke models using the PubMed, Scopus, and Google Scholar databases up to December 2024. A total of 91 articles were identified, of which six met the eligibility criteria. Data were extracted from six studies (three in vivo and three in vitro), and their therapeutic outcomes are summarized in tables. All included studies consistently demonstrated beneficial effects of hypMSC-CM, with findings of reduced infarct volume, improved neurological function, and the promotion of neurogenesis and angiogenesis. This systematic review suggests that hypMSC-CM enhances neurogenesis and neuroprotection in the brain. However, high-quality preclinical studies and clinical trials are needed to support the therapeutic benefits of hypMSC-CM for ischemic stroke, especially as an adjuvant to standard tPA therapy.

Background and Aim: Angiogenesis and anti-apoptosis play crucial roles in ischemic stroke recovery. The mesenchymal stem cell (MSC) secretome, rich in bioactive molecules, presents a promising therapeutic avenue. However, optimizing the culture conditions to enhance the expression of angiogenic markers remains a challenge. This study examines the effect of hypoxic preconditioning on the expression of vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinase-2 (MMP-2), and microRNA (miRNA-128) in the secretome of umbilical cordderived MSCs (UC-MSCs) from Macaca fascicularis. Materials and Methods: UC-MSCs were cultured under normoxic (21% O2 ) and hypoxic conditions (1%, 3%, and 5% O2 ) for 48 h. The secretome was isolated, and reverse transcription-quantitative polymerase chain reaction was used to quantify the expression of VEGF, MCP-1, MMP-2, and miRNA-128. Expression levels were normalized to housekeeping genes and analyzed using statistical methods to determine significant differences among groups.Results: Hypoxic preconditioning significantly upregulated VEGF (1% O2 ), MCP-1 (5% O2 ), and miRNA-128 (5% O2 ) expression compared to normoxic conditions. Conversely, MMP-2 expression was highest in normoxic conditions and downregulated under hypoxia. In addition, miRNA-128 was found to be predominantly secreted into the extracellular space under hypoxic conditions rather than retained within cells. Conclusion: Hypoxic preconditioning effectively modulates the expression of key angiogenesis and anti-apoptotic markers in UC-MSCs. The study highlights the importance of optimizing oxygen levels to enhance the therapeutic potential of MSCderived secretomes for ischemic stroke treatment. Future research should focus on in vivo validation and clinical translation of these findings.

Background: Primary cells have the same heterogeneity and differentiation capacity that has potential as an in vitro ischemic stroke models. It is hoped that primary cells from non-human primates/NHPs that have genetic similarities to humans can provide molecular information and become more accurate data for use in drug screening, especially stroke therapy. B27 is a supplement commonly used in neuronal cell cultures, but there are concerns that its effects will interfere with the neuroprotective processes of the drug candidates being tested. Aim: This research will prove the demonstrate of neurons as a ischemic stroke model and the effects of B27 in Macaca fascicularis (Mf) neurons as a model for ischemic stroke under oxygen glucose deprivation/OGD. Methods: Neurons were obtained from a collection of biological materials collected during previous research. Neuronal validation was performed using immunocytochemistry/ICC with the marker β-tubulin. Expression of the apoptotic response was performed by RT-PCR using Bax, BCL-2, caspase-9, and p53 gene markers. Characterization of neurons in terms of positive tβubulin markers and induction of OGD in neurons can be performed for 6 h to model ischemic stroke. Results: This study showed that cultured neurons under OGD conditions can experience apoptosis, namely by increasing pro-apoptosis and decreasing anti-apoptosis. However, B27 supplementation increased the expression of anti-apoptotic Bcl-2 genes and decreased proapoptotic genes such as Bax, caspase 9, and p53. Conclusion: Neuron culture from Mf can be used as an in vitro model of ischemic stroke, and B27 supplementation in neurons exerts neuroprotective effects on the induction of OGD.

The PI3K/Akt pathway plays a crucial role in regulating a broad network of proteins involved in the proliferation of HER2-positive breast cancer. The ethyl acetate fraction of Vernonia amygdalina, which contains cardiac glycosides, has been shown to reduce the expression of PI3K and mTOR. However, the specific cardiac glycoside compounds with significant potential as PI3K inhibitors have yet to be clearly identified. This study employs machine learning to perform virtual screening of cardiac glycosides from V. amygdalina against the p110 subunit of PI3K. Initially, Lipinski’s Rule of Five was used to filter the PIK3CA inhibitor database via KNIME software. Subsequently, QSAR modeling was conducted using KNIME’s machine learning platform, employing six different algorithms. Cardiac glycosides from V. amygdalina were then evaluated using the best-performing QSAR model. The top three compounds identified underwent molecular docking and molecular dynamics simulations. The random forest algorithm was selected as the primary predictive model, which identified Vernoamyosides A (VG-1), Vernoniamyosides D (VG-8), and Vernoniosides A4 (VG-10) as the compounds with the highest confidence levels. Molecular docking results indicated that these three compounds exhibited stronger and more stable interactions with the PIK3CA receptor compared to alpelisib, a known PIK3CA inhibitor. Furthermore, molecular dynamics simulations revealed that VG-10 had the lowest binding free energy, as determined by MM-GBSA analysis. The findings of this study provide a foundational basis for preclinical and clinical investigations aimed at developing PI3K inhibitors derived from cardiac glycosides of V. amygdalina for the treatment of HER2+ breast cancer.

Background:Primary cells have the same heterogeneity and differentiation capacity that has potential as an in vitro ischemic stroke models. It is hoped that primary cells from non-human primates that have genetic similarities to humans can provide molecular information and become more accurate data for use in drug screening, especially stroke therapy. B27 is a supplement commonly used in neuronal cell cultures, but there are concerns that its effects will interfere with the neuroprotective processes of the drug candidates being tested. Aim:This research will prove the demonstration of neurons as an ischemic stroke model and the effects of B27 in Macaca fascicularis (Mf) neurons as a model for ischemic stroke under oxygen glucose deprivation (OGD). Methods:Neurons were obtained from a collection of biological materials collected during previous research. Neuronal validation was performed using immunocytochemistry with the marker β-tubulin. Expression of the apoptotic response was performed by real time- polymerase chain reaction/(RT-PCR) using Bax, BCL-2, caspase-9, and p53 gene markers. Characterization of neurons in terms of positive tβubulin markers and induction of OGD in neurons can be performed for 6 h to model ischemic stroke. Results:This study showed that cultured neurons under OGD conditions can experience apoptosis, namely by increasing pro-apoptosis and decreasing anti-apoptosis. However, B27 supplementation increased the expression of anti-apoptotic Bcl-2 genes and decreased proapoptotic genes such as Bax, caspase 9, and p53. Conclusion:Neuron culture from Mf can be used as an in vitro model of ischemic stroke, and B27 supplementation in neurons exerts neuroprotective effects on the induction of OGD.

Mesenchymal Stem Cell (MSC) secretome has potential as a neuroprotective and neuroregenerative agent. It can have effects due to its paracrine factors, such as Brain Derived Neurotrophic Factor (BDNF) and Stromal-Cell Derived Factor-1 (SDF-1) which can be induced with hypoxia preconditioning. This compound may play a role in the treatment of neurological diseases. Stroke has become a neurological disease that contributes to high rates of mortality and morbidity worldwide. There have been several pre-clinical trials on animal stroke models using MSC secretomes from rats and humans, but no studies have been conducted on Non-Human Primate, such as Macaca fascicularis. This species has been widely used in biomedical research and part of it can be utilized for such studies which will reduce the cost of using human MSC. The results of this study, Umbilical Cord (UC)-MSCs of Macaca fascicularis have been successfully cultured and characterized in terms of phenotypic and differentiation. Hypoxia precondition was able to induce BDNF secretion up to 264 pg/mL and SDF-1 up to 666 pg/mL in the UCMSC secretome. Hypoxic preconditioning with 3% oxygen can induce the most optimal BDNF and SDF-1 secretion, compared to 1% and 5% hypoxia.

Vernonia amygdalina Delile, coming from the Asteraceae tribe, contains active compounds that can treat breast cancer. This study examines the anticancer activity of Vernonia amygdalina Delile leaves, an active fraction of MCF-7/HER-2 breast cancer cells. Thin-layer chromatography determines the phytochemical screening. Cytotoxic and proliferation analyses were determined using the MTT method for the MCF-7/HER-2 breast cancer cell line. The cell cycle and apoptosis profiles were examined using flow cytometry. The results can be summarized in this study: Vernonia amygdalina Delile fraction contains a flavonoid with great potential for pharmacological activities, for instance, inhibiting the growth of cancer cells. The ethyl acetate fraction was more potent and cytotoxic on MCF-7/HER-2 cells (IC50 = 66 μg/mL) than extract ethanol (IC50 = 130 μg/mL). The ethylacetate fraction of Vernonia amygdalina Delile has been proven to inhibit cell proliferation by decreasing cell viability with an IC50 of 66 μg/mL concentration incubated for 24, 48, and 72 hrs with cell viability values of 64.46%, 61.67%, and 53.89%, respectively, compared to the ethanol extract IC50 of 130 μg/mL concentration with cell viability values of 56.0%, 50.19%, and 58.67%, respectively, which induced apoptosis and inhibited the cell cycle in the G2-M phase. To conclude, the ethyl acetate fraction of Vernonia amygdalina Delile can be used as an anticancer against the MCF-7/HER-2 cell line with an IC50 of 66 μg/mL, inhibiting cell proliferation and the cell cycle and inducing apoptosis activity in the MCF-7/HER-2 cell.

OBJECTIVE:This study is targeted at assessing the chemotherapy factor of the ethanol extract of Vernonia amygdalina Delile (VAD). METHODS:U87 glioblastoma cells were treated with extract and a fraction of Vernonia amygdalina Delile (VAD) was harvested from the herbarium. Cytotoxicity was evaluated to determine the IC50 through microscopic observation followed by an MTT assay. Subsequently, flow cytometry with a FACS type was employed to conduct cell cycle and apoptosis analyses. Annexin V/PI and PI markers were used to assess apoptosis and cell cycle progression. RESULT:The ethanol extract and ethyl acetate fraction of VAD showed promising effects as cancer chemotherapy in glioblastoma cells. The IC50 values for the extract and fraction were notably low, at 37.65 µg/ml and 10.12 µg/ml, respectively, for U87 cells. Analysis of apoptosis using FACS revealed a more pronounced apoptotic effect of the 15 µg/ml fraction of VAD on both early and late apoptosis compared to the 75 µg/ml extract of VAD. Although some differences in cell cycle properties were observed, there were no significant differences in cell cycle analysis between the extract and fraction. CONCLUSION:These findings underscore the efficacy of VAD's ethanol extract and ethyl acetate fraction as chemotherapeutic agents against U87 cancer cells. The low IC50 values and significant induction of early apoptosis highlight the cytotoxic effects of these treatments on U87 cells.

OBJECTIVE:Glioblastoma Multiforme (GBM) poses a significant challenge due to its high aggressiveness and unfavorable prognosis, with existing treatments demonstrating limited efficacy in prolonging survival rates. This study aimed to assess the anticancer properties of Aaptos suberitoides extracts and fraction on the U87 cell line, serving as a representative model for GBM. METHODS:U87 cells were treated with ethanol extracts derived from Aaptos suberitoides, specifically two extracts (OAA-1 and OAA-2) and one ethyl acetate fraction (EA) isolated from specimens collected on Pramuka Island and Tinjil Island. The evaluation encompased microscopic observation and MTT assay to determine the IC50. Subsequently, antiproliferative effects were investigated through apoptosis and cell cycle assays. RESULTS:The extract demonstrated cytotoxic activity against U87 cells, with OAA-1 and OAA-2 exhibiting IC50 values of 35.78 μg/mL and 25.38 μg/mL, respectively. OAA-1 notably induced apoptosis at 50 μg/mL and induced cell cycle arrest. On other hand, OAA-2, while also inducing apoptosis significantly, had a lesser impact on cell cycle arrest. In contrast, EA induced significant apoptosis at a concentration of 100 μg/mL. CONCLUSION:The ethanol extracts and the ethyl acetate fraction of Aaptos suberitoides emerged as a promising candidate for Glioblastoma Multiforme cancer therapy, showing potential in inhibiting cell proliferation and inducing apoptosis.

Vernonia amygdalina Delile or bitter leaf, a traditional medicinal plant endemic to Africa, has shown anticancer properties by inhibiting the downstream signaling of human epidermal growth factor receptor 2 (HER2) in several cancer cells through the PI3K/AKT/mTOR pathways. This study explored the potential target of Vemonia cardiac glycosides for the treatment of HER2+ breast cancer (CTG) using bioinformatics and in vitro experiments. The similarity ensemble technique Swiss TargetPrediction was used to predict protein targets of cardiac glycosides. DisGeNET and UALCAN were used to collect the regulatory genes of HER2+ breast cancer cells. We obtained 75 CTG sequences from the predicted target and regulatory genes of HER2+ breast cancer. Subsequent analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment revealed that CTG was enriched for multiple pathways, such as PI3K-Akt signaling, breast cancer, EGFR tyrosine kinase inhibitor resistance, and ErbB signaling pathways. Further bioinformatic analysis identified five CTG: ERBB2, ESR1, EGFR, PIK3CA, and PTPN11. The ethanolic extract and its ethyl acetate fraction of V. amigdalina were referred to as REF and EA, respectively. Cytotoxicity tests showed that EA exerted strong cytotoxicity on MCF-7/HER2 cells by inducing S-phase cell cycle arrest and apoptosis, modulating the mRNA expression of CTG: ERBB2, ESR1, and proliferation and apoptosis markers, PCNA, CCND1, BAX, BCL2, and CASP7. Future studies should develop EA and cardiac glycosides as anticancer agents against HER2+breast cancer cells. (c) 2023 SAAB. Published by Elsevier B.V. All rights reserved.

BACKGROUND:3-dimensional (3D) printing carries a genuine potential for pre-operative planning in neurosurgery. Entry-level 3D printers offer practicality in low resource settings, but are often limited by the range of filament materials as well as the capability of open-source segmentation software.OBJECTIVE:We intended to demonstrate that 3D printing of neuroanatomical structures is possible using an entry-level 3D printer equipped with the direct drive (DD) modification, which supported flexible filaments, with the models segmented using an open-source software.METHODS:A DD system was installed onto the Ender 3 Pro printer. An attempt to print neurosurgical models using a low-cost 3D printer was conducted, where four patient-based neuroanatomical models were printed: skull base-vasculature, skull base-tumour, cervical spine, and ventricular system. The results were discussed and compared to similar endeavours in past literature.RESULTS:Although DD installation was challenging and led to vibration and longer print time, which ultimately warranted an inferior printing speed, DD system enabled the printing with thermoplastic polyurethane (TPU), a versatile elastomer; in addition to producing equal amount of detail to those printed with high-end printers and advanced image segmentation software. Fitting the frame well, changing infill type, and avoiding warping and stringing will improve print quality with the DD system.CONCLUSION:3D printing with entry-level 3D printers equipped with DD system has been proven to be a reliable way of accurately reproducing patient-specific neuroanatomical constructs. Follow-up studies are necessary to implement 3D printing for neurosurgical planning in low-resource settings.

Background:The occurrence of spinal fracture due to tetanus nowadays is extremely rare, as compared to the 1950s, since the widely available anti-tetanus and antispasmodic therapy. The spinal fracture in tetanus patients is usually reported in higher thoracic vertebrae, previously with a rate as high as 57.5%. Spondylitis is the most common form of skeletal tuberculosis (TB) and can cause a spinal fracture. In Indonesia, tetanus is still reported, while tuberculosis is still endemic; however, co-infection of both diseases is rarely reported.Case Presentation:A 36-year-old male was brought to our hospital with jaw stiffness, accompanied by fever. A history of dental cavities was present, and 5 days prior, he experienced a fishing hook wound on his right index finger. There was no history of TB. Physical examination showed meningismus, 2 cm trismus, abdominal spasm, opisthotonus, and spontaneous muscle spasms, without dysautonomia. In the third week of hospitalization, while his tetanus condition improved, he complained of weakness in both legs. A thorough history taking revealed a history of backache for 3 years. A wedge-shaped fracture on his 11th and 12th thoracic vertebrae was observed on radiographic examination. A spinal TB diagnosis was made, and treatment was started. He refused to get spinal surgery, then went home with 4 out of 5 motor strength scale. After three months, he returned to his routine activity as a food hawker with no motor deficits.Conclusion:Tetanus spinal fracture is extremely rare nowadays; a thorough history of spinal problems/medication is compulsory for anticipation. This patient's spinal fracture was deemed due to a preexisting TB spinal infection that was precipitated by prolonged continuous tetanic spasm due to general tetanus.

Introduction:Spinocerebellar ataxia type-3 (SCA3) is an adult-onset autosomal dominant neurodegenerative disease. It is caused by expanding of CAG repeat in ATXN3 gene that later on would affect brain structures. This brain changes could be evaluated using brain MRI volumetric. However, findings across published brain volumetric studies have been inconsistent. Here, we report MRI brain volumetric analysis in a family of SCA 3 patients, which included pre-symptomatic and symptomatic patients.Methodology:The study included affected and unaffected members from a large six-generation family of SCA 3, genetically confirmed using PolyQ/CAG repeat expansion analysis, Sanger sequencing, and PCR. Clinical evaluation was performed using Scale for the Assessment and Rating of Ataxia (SARA). Subjects' brains were scanned using 3.0-T MRI with a 3D T1 BRAVO sequence. Evaluations were performed by 2 independent neuroradiologists. An automated volumetric analysis was performed using FreeSurfer and CERES (for the cerebellum).Result:We evaluated 7 subjects from this SCA3 family, including 3 subjects with SCA3 and 4 unaffected subjects. The volumetric evaluation revealed smaller brain volumes (p < 0.05) in the corpus callosum, cerebellar volume of lobules I-II, lobule IV, lobule VIIB and lobule IX; and in cerebellar gray matter volume of lobule IV, and VIIIA; in the pathologic/expanded CAG repeat group (SCA3).Conclusion:Brain MRI volumetry of SCA3 subjects showed smaller brain volumes in multiple brain regions including the corpus callosum and gray matter volumes of several cerebellar lobules.

Spinocerebellar ataxia (SCA) 3 is a neurodegenerative disease which involves cerebellum and extra cerebellum. Neuropathy in SCA3 manifests in various ways, including axonal and demyelination lesions in sensory and motor nerves. There has not been any study that describes the peripheral neuropathy characteristics of SCA3 patients in Indonesia at the time of this publication. This paper reports a case of a 43-year-old male with known spinocerebellar ataxia 3 presented with hereditary ataxia and mild numbness in both palms since two years before. No abnormalities were found during the sensory examination. The NCS showed severe axonal demyelinating sensorimotor peripheral neuropathy. In magnetic resonance imaging (MRI), an atrophy in the cerebellum with cerebral multiple lacunar infarction was identified. Electrophysiological results revealed profound axonal lesion in peripheral nerves. To conclude, peripheral neuropathy in SCA3 represents the dominance of axonal lesions in motor nerves.

OBJECTIVE:This study aimed to investigate the cytotoxicity, anti-proliferation and anti-migration effect of the ethanol extract of Aaptos suberitoides on trastuzumab-resistant HER2+ breast cancer cell line. METHODS:Aaptos suberitoides was collected from Tinjil Island, Banten, Indonesia, and was processed with maceration and ethanol extraction. HCC-1954 cells were treated with the ethanol extract and then followed by 3- [4, 5-dimethylthiazol-2-yl] -2.5 diphenyl tetrazolium bromide (MTT) assay to assess cytotoxicity, clonogenic assay and three-dimensional (3D) spheroid assay to evaluate anti-proliferative effect in two-dimensional and 3D model, respectively, and wound healing assay to determine anti-cell migration effect. Four parametric regression was used to analyse the IC50. RESULTS:This study revealed that the ethanol extract of Aaptos suberitoides suppressed cell viability in correlation with cell death induction. The IC50 values of the ethanol extract of Aaptos suberitoides using MTT assay and clonogenic assay were 12.0 ppm and 4.36 ppm, respectively. The extract demonstrated an inhibition effect on spheroid growth. In low concentration, the extract of Aaptos suberitoides inhibited cell migration. Furthermore, MS analysis showed that the most abundant compounds in this extract has molecular weight m/z 229.81 [M+H]+. CONCLUSION:This study revealed that the ethanol extract of Aaptos suberitoides demonstrates cytotoxicity, anti-proliferation and anti-migration effect as well as inhibition effect on three-dimensional spheroid growth in trastuzumab-resistant HER2+ breast cancer cell line..

Spinocerebellar ataxia (SCA) is an autosomal dominant hereditary disease with progressive course, and no causal therapy. Diagnostics are still challenging, due to facility and protocols, and so as in Indonesia. As a national referral center, Dr. Hasan Sadikin Central General Hospital has received a lot of patients from all over Indonesia, particularly from Western Java. Study related to SCA (including clinical and genetic profile) is still limited in Indonesia. We identified index patients from three families with ataxia, hence intend to determine their clinical and genetic pattern. The hereditary pattern is autosomal dominant. Scale for the assessment and rating of ataxia (SARA) shows mild and moderate ataxia. Inventory of non-ataxia signs (INAS) scores of the patients were 3, 5 and 6. Montreal cognitive assessment-Indonesian version (MOCA-INA) shows only one patient has mild cognitive impairment, despite young age. Barthel index shows 1 subject has moderate dependency. Mutation in Ataxin3 polyQ repeats shows pathologically long CAG repeats, 72,10; 72,10; and 72,23 respectively in mutant and wild type allele. We diagnosed the index patients with spinocerebellar ataxia type 3. This study is the first case series study in Indonesia. The hereditary pattern is clearly shown as an autosomal dominant ataxia. The clinical and genetic profile was varied, and the symptom is progressive and deteriorates overtime, including wide based gait, speech problem, motor and sensor complaint, and cognitive decline complaint. Despite the same polyQ stretch length, the onset and clinical characteristics of patients are diverse.

Breast Cancer (BC) is the leading cause of cancer death in women. One BC subtype is very aggressive with amplification of human epidermal growth factor receptor 2 (HER2) protein. Although specific HER2+ targeting agents are available, most of HER2+ BC patients develop resistant to these agents. Recent studies show that metformin, is able to become anti-tumor in various cancer cells. This research aims to evaluate anti-tumor activities of metformin to HER2+ BC cells in both sensitive and resistant to trastuzumab. A series of assays were performed to evaluate metformin anti-tumor activities in HCC-1954 and SKBR-3 HER2+ BC cells. MTT assay was performed to evaluate cell death, and inhibitory concentration (IC50), while scratch assay was performed to assess inhibition of cell migration and clonogenic assay to assess cell proliferation. p<0.05 was considered to be significant. Metformin could suppress the number of HER2+ BC cells. Viability assay showed suppression of viable cells after metformin incubation of 60 and 600 µM compared to control, 30 and 90%, respectively. Surprisingly, IC50 of metformin was smaller in HER2+ BC HCC-1954 cells that resistant to trastuzumab compare to the sensitive one (SKBR-3). Both were below 1 µM, with R2 more than 0.95. Additionally, clonogenic assay showed less colony number and colony area with at least p < 0.05 in colony number and p < 0.01 in the area. In addition, metformin inhibited cell migration of HER2+ BC cells. Metformin shows a potency as anti-tumor by inducing cell death, inhibiting cell proliferation and cell migration of HER2+ BC cells.

Breast cancer (BC) and cervical cancer (CC) have a high prevalence and mortality rate worldwide. Despite the availability of advanced treatment, resistance to conventional chemotherapies has emerged. Myrmecodia pendens, one of the species of Sarang Semut (local name), possess a potential of antitumor effects by inducing cell death different cancer cell entities. This study aimed to assess anti-tumor activities of n-hexane fraction of M. pendens in inhibiting cell survival and cell migration in BC and CC cells. M. pendens was extracted in methanol then fractionated using n-hexane or ethyl acetate. BC cells including MCF-7 (luminal A), HCC-1954 (HER2+) cells and CC Hela cells were treated with M. pendens extracts to evaluate cytotoxic activity using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay as well as anti-cell migration using scratch assay. We also analyzed inhibitory concentration 50 (IC50) of n-hexane fraction in BC and CC cells. We started with comparing cytotoxicity activities of methanol extract, ethyl acetate and n-hexane fractions of M. pendens. Data showed that the n-hexane fraction was the most potent inducing BC cell death. Therefore, we used the n-hexane fraction for further experiments. Interestingly, IC50 of this fraction in HCC-1954 and Hela cells were lower than in MCF-7 cells, 16; 13 and 60 ppm, respectively. Moreover, the low concentrations of n-hexane fraction inhibited HeLa cells migration, compared to control group (p<0.05). The n-hexane fraction of M. pendens shows promising anti-cancer agent, by inhibiting BC and CC cell survival as well as inhibiting CC cells migration. Keywords: breast cancer, cervical cancer, MTT assay, Sarang Semut, scratch assay