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Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate Putu Juniartha; Muhammad Rasjad Indra; Hidayat Sujuti; Diana Lyrawati; Tatit Nurseta
Journal of Tropical Life Science Vol. 6 No. 3 (2016)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.06.03.11

Abstract

Cervical cancer recurs in 90% cases and linked to cancer stem cells that able to self-renew and responsible for recurrence, metastasis, and mortality of cancer. Isolation and identification of cancer stem cells using serum-free medium needs expensive growth factors and consume time. This study try to grow tumor sphere using culture plate coated with 1% agarose as an efficient and economical alternative to non-adhesive culture plate. HeLa cell line was grew in culture plate coated with 1% agarose and non-adhesive culture plate using similar medium and culture condition. Tumor spheres morphology was observed and the colonies were counted in 7 days followed by single cell assay. Tumor spheres then counted for CD133+, CD34+, and Sox2 expression using flowcytometry. Culture plate coated with 1% agarose can be used as an economic and efficient alternative to culture tumor sphere. Using culture plate coated with 1% agarose, the tumor spheres formed in 7 days with similar morphology to non-adhesive culture plate. Tumorsphere had three dimensional – sphere shape that tightly attached, colonized, and overlapped. The tumor sphere colony counts of two plates were statistically have no significant difference (p=0,667). Single cell assay of a tumor sphere shows that it can grow new tumor spheres with similar morphology. The tumor sphere from culture plate coated with 1% agarose express CD133+ and CD34+ as much as 8.78% ± 2.14 and Sox2 as much as 35.30% ± 23.82 whereas tumor sphere from non-adhesive culture plate express CD133+ and CD34+ as much as 62.36% ± 1.06 and Sox2 as much as 98.86% ± 0.56 (p = 0000).
Differences in the Levels of Interleukin-1β (IL-1β) and Interleukin-1 Receptor Antagonist (IL-1ra) in Children with Status Epilepticus and Febrile Seizure Fita Shofiyah; Masdar Muid; Hidayat Sujuti
Journal of Tropical Life Science Vol. 7 No. 3 (2017)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.07.03.05

Abstract

Proinflammatory cytokines are elevated in status epilepticus and febrile seizure and associated with tissue damage. This study aimed to investigate the differences in interleukin 1 beta (IL-1β) and interleukin 1 receptor agonist (IL-1ra) levels in status epilepticus as compared with febrile seizure and febrile. This cross-sectional study was designed to include 45 subjects divided equally into three groups (status epilepticus, febrile seizure, and febrile). Both IL-1β and IL-1ra were measured by using an ELISA method. Results showed that IL-1β levels were significantly higher in the status epilepticus group as compared with the febrile seizure and febrile groups (p < 0.05). IL-1ra levels in the status epilepticus group were significantly lower compared with the febrile seizure group (p = 0.04). Consistently, the IL-1β/IL-1ra ratio in the status epilepticus group was significantly higher as compared with the febrile seizure group (p = 0.01). We concluded that IL-1β and the IL-1β/IL-1ra ratio were significantly higher in status epilepticus. IL-1ra levels were significantly higher in the febrile seizure group.
Immunohistochemistry Evaluation of TGF-β1, SOX-9, Type II Collagen and Aggrecan in Cartilage Lesions Treated with Conditioned Medium of Umbilical Cord Mesencyhmal Stem Cells in Wistar Mice (Rattus novergicus) Bintang Soetjahjo; Mohammad Hidayat; Hidayat Sujuti; Yuda Fibrianto
Journal of Tropical Life Science Vol. 8 No. 1 (2018)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.08.01.05

Abstract

Currently, umbilical cord mesenchymal stem cells have the potential to be used as treatment options for any cartilage lesion. This research aimed to evaluate the effects of conditioned medium from umbilical cord mesenchymal stem cells (UC-MSC) on damaged cartilage through the expression of proteins TGF-β1, SOX-9, type II collagen and aggrecan, which are known to be related to chondrogenesis. UC-MSC were isolated from 19-days-pregnant Wistar mice and were cultured using the standard procedure to obtain 80% confluence. Subsequently, the culture was confirmed through a microscopic examination that was driven to be an embryoid body to obtain a pre-condition medium. This research utilized 3-month-old male Wistar mice and was categorized into 6 groups (3 control and 3 treatment groups). Each animal had surgery performed to create a femur condyle cartilage defect. The treatment groups were administered a dose of stem cells at 1 mL/kg. Next, immunohistochemical (IHC) staining was performed to examine the expression of TGF-β1, SOX-9, type II collagen and aggrecan in the 2nd, 3rd, and 4th month of evaluation. The results were analyzed statistically using ANOVA test. For each of the treatment groups, there was increased expression (p < 0.05) in all proteins TGF-β1, SOX-9, type II collagen and aggrecan when compared with control groups at the 2nd, 3rd, and 4th month of evaluation. Pre-conditioned medium from UC-MSC potentially increases the expression of TGF-β1, SOX-9, type II collagen and aggrecan in the damaged cartilage of Wistar mice.
Correlation of Malondialdehyde (MDA) and C-reactive Protein (CRP) Level to Neurodevelopmental Outcome in Children After the Episode of Convulsive Type Status Epilepticus Sintha Restuningwiyani; Ariani Ariani; Hidayat Sujuti; Masruroh Rahayu; Krisni Subandiyah
Journal of Tropical Life Science Vol. 8 No. 3 (2018)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.08.03.07

Abstract

Refractory and mortality associated with status epilepticus (SE) were correlated with the degree of inflammation-induced neuronal cell death. This study was aimed to investigate the correlation of oxidative stress (Malondialdehyde, MDA) and inflammation (C-reactive protein, CRP) process with neurodevelopmental outcome in children after the episode of convulsive type SE. This study was designed as cross sectional which included 26 convulsive type SE subjects and 15 control subjects. MDA level was measured by thiobarbituric acid (TBA) method, while CRP level was measured by ELISA method. Neurodevelopmental outcome was measured by Bayley-III Scale of Infant and Toddler Development 3 month after the convulsive type SE episode. Results showed that both MDA (independent t-test, p < 0.05) and CRP (Mann-Whitney test, p < 0.05) level was significantly higher in convulsive type SE group as compared to control group. Further analysis also showed that MDA (Spearman correlation test, p = 0.000, r = 0.756) and CRP (Spearman correlation test, p = 0.000, r = 0.835) level was positively correlated with convulsive type SE. In convulsive type SE group, MDA level was negatively correlated with neurodevelopmental outcome but CRP was not. We concluded that MDA level was negatively correlated with neurodevelopmental outcome in children with convulsive type SE, but CRP was not.
Sensitivity and Specificity of Nested PCR for Diagnosing Malaria: Cases in Several Areas of Indonesia Samsul Arifin; Loeki Enggar Fitri; Hidayat Sujuti; Bagus Hermansyah; Agustina Tri Endharti; Niniek Burhan; Didi Candradikusuma; Erma Sulistyaningsih; Josef Sem Berth Tuda; Umar Zein
Journal of Tropical Life Science Vol. 8 No. 2 (2018)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.08.02.11

Abstract

Indonesia is still included in high endemic area of malaria infection. Early detection as well as appropriate and quick treatment is needed to be able to prevent and treat malaria in Indonesia. Laboratory examination using a microscopic method is still used as the gold standard to diagnose malaria cases. However, the morphology similarity of some Plasmodium species and the number of parasites that can be seen under microscopy causes malaria diagnosis become difficult if only relying on microscopy diagnostic method. The purpose of this study is to analyze the sensitivity and specificity of nested PCR compared to microscopic examination in diagnosing malaria cases. A cross-sectional study has been carried out in some areas of Indonesia and the microscopic analysis as well as nest PCR was done in Laboratory of Parasitology and Laboratory of Central Biomedical Faculty of Medicine, Universitas Brawijaya, Malang East Java Indonesia. A total of 149 blood samples from patients with clinical symptoms of malaria had been obtained from Sumatra, Sulawesi and East Java during December 2011 to December 2013. From 149 sample, 81.9% samples were diagnosed malaria positive by microscopy examination, whereas the PCR results showed that 90.6% of samples were positive. Nested PCR sensitivity is 97.5%, and microscopy 88.2%. Nested PCR specificity is 40.7%, whereas microscopy 78.5%. PPV and NPV for nested PCR are 88,2% and 78.5% respectively, and for microscopy are 97.5% and 40.7% respectively. Nested PCR has a higher sensitivity than microscopy in diagnosing malaria and is able to detect mixed infection better than microscopic examination. However, it is statistically less specific than microscopy examination.
Possible Role of Akt in Mossy Fiber Sprouting: Akt Activity and CA3 Mossy Fiber Sprouting in a Kainate Model of Epilepsy: Akt Activity and CA3 Mossy Fiber Sprouting in Epilepsy husna, Machlusil; Handono, Kusworini; Sujuti, Hidayat; Aulanni'am, Aulanni'am
Journal of Tropical Life Science Vol. 14 No. 3 (2024): In Press
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.14.03.09

Abstract

The most prevalent pathological phenomenon observed in patients with epilepsy is hippocampal mossy fiber sprouting (MFS), which is thought to be associated with epileptic progression, such as worsening seizure control, cognitive function, and behavior. MFS is discovered in the dentate gyrus and the hippocampal Cornu Ammon 3 (CA3) area. The CA3 area is involved in memory, so disturbances in that area can affect memory impairment in patients with epilepsy. The mammalian target of rapamycin complex-1 (mTORC1) is also associated with MFS. Akt is an upstream activator of mTORC1 and a downstream target of mammalian target of rapamycin complex-2 (mTORC2) and plays a role in cytoskeleton organization. We analyzed Akt activity and MFS in the CA3 zone in an in vitro model of kainate-induced epilepsy. We divided organotypic hippocampal slice cultures (OHSCs) into a kainate (epilepsy) group and a control (untreated) group. On the 10th day in vitro (DIV), the kainate group was exposed to 8 µM kainic acid for 48 h, diluted in the medium. At 32 DIV, we measured Akt activity through western blotting and CA3 MFS through synaptoporin fluorescence intensity observed using confocal laser scanning microscopy. We found that Akt activity increased significantly (p = 0.000) in the kainate group, and the synaptoporin fluorescence intensity also increased in the stratum oriens of the CA3 area (p = 0.049) in the kainate group. Our findings implied that Akt may play a role in MFS development. Because Akt is a main downstream target of mTORC2, mTORC2 may also be involved in MFS development. Further research is required to clarify these findings.
Possible Role of Akt in Mossy Fiber Sprouting: Akt Activity and CA3 Mossy Fiber Sprouting in a Kainate Model of Epilepsy: Akt Activity and CA3 Mossy Fiber Sprouting in Epilepsy husna, Machlusil; Handono, Kusworini; Sujuti, Hidayat; Aulanni'am, Aulanni'am
Journal of Tropical Life Science Vol. 14 No. 3 (2024)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.14.03.09

Abstract

The most prevalent pathological phenomenon observed in patients with epilepsy is hippocampal mossy fiber sprouting (MFS), which is thought to be associated with epileptic progression, such as worsening seizure control, cognitive function, and behavior. MFS is discovered in the dentate gyrus and the hippocampal Cornu Ammon 3 (CA3) area. The CA3 area is involved in memory, so disturbances in that area can affect memory impairment in patients with epilepsy. The mammalian target of rapamycin complex-1 (mTORC1) is also associated with MFS. Akt is an upstream activator of mTORC1 and a downstream target of mammalian target of rapamycin complex-2 (mTORC2) and plays a role in cytoskeleton organization. We analyzed Akt activity and MFS in the CA3 zone in an in vitro model of kainate-induced epilepsy. We divided organotypic hippocampal slice cultures (OHSCs) into a kainate (epilepsy) group and a control (untreated) group. On the 10th day in vitro (DIV), the kainate group was exposed to 8 µM kainic acid for 48 h, diluted in the medium. At 32 DIV, we measured Akt activity through western blotting and CA3 MFS through synaptoporin fluorescence intensity observed using confocal laser scanning microscopy. We found that Akt activity increased significantly (p = 0.000) in the kainate group, and the synaptoporin fluorescence intensity also increased in the stratum oriens of the CA3 area (p = 0.049) in the kainate group. Our findings implied that Akt may play a role in MFS development. Because Akt is a main downstream target of mTORC2, mTORC2 may also be involved in MFS development. Further research is required to clarify these findings.
Co-Authors A. Harijono A.A. Ketut Agung Cahyawan W Abdalrauf A Mahmud Yousif Abdalrauf A Mahmud Yousif, Abdalrauf A Mahmud Abdussalam Ashour, Abdussalam Achmad Rudijanto Adinda Chika Anindita Affandi, Haykal Agung Prasetyo Wibowo Agustiana Dwi Agustiana Dwi Indiah Ventiyaningsih Agustina Tri Endharti Ala Shukri Eshami, Ala Shukri Analis W Wardhana Anik Puryatni Anisak, Siti Ariani Ariani Ariani Ariani Arifin, Mochammad Samsul Arsinah Habibah Fitriah, Arsinah Habibah Askandar Tjokroprawiro Astika Swastirani Aulanni'am, Aulanni'am Aulia Yasmin, Aulia Bagus Hermansyah Bambang Rahardjo Bayu Kurniawan Bintang Soetjahjo Bogi Pratomo Wibowo Burhan, Niniek Chabibi, Mochamad Chabibi Chairinnisa, Elsa Safira Christina Dewi Dalhar, Mochamad Dewi, Ni Ketut Ayu Feriyanti Dian Handayani Dian Nugrahenny Diana Lyrawati Diana Lyrawati Dicky Faizal Irnandi Didi Candradikusuma Didi Candradikusuma Dwi Jayasa, Pande Made Dwi Yuni Nur Hidayati Edi Mustamsir Edi Widjajanto Edi Widjajanto Edwin Widodo Effendi, Ma'sum Eko Sulistijono Elok Zubaidah Erma Sulistyaningsih Ery Olivianto Fardizia Putri Alia Feni Istikharoh Fibrianto, Yuda Fita Shofiyah Gading, Inges Manggar Gerry Gunawan, Gerry Handono Kalim Hanif Hanif Happy Kurnia Permatasari Hara, Kenta HARI PURNOMO Hariwati Hariwati Hasfiani, Yuliatin Huda Rohmawati Husna, Ully Husnul Khotimah Ika Kustiyah Oktaviyanti Indrayanti Indrayanti Indriati Dwi Rahayu Inggita Kusumastuty Josef Sem Berth Tuda Juniartha, Putu Kalsumy, Umi Kanthi Permaningtyas Tritisari, Kanthi Permaningtyas Karyono Mintaroem Karyono Mintaroem Karyono Mintaroem Karyono mitaroem, Karyono Krisni Subandiyah Krisni Subandiyah Kusworini Handono Laily Yuliatun Lavina Sofia Ardani Lely Retno W Leny Budhi Harti Lestari, Bayu Lirawati, Diana Loeki Enggar Fitri M Rasjad Indra Machlusil Husna, Machlusil Maimun Z Arthamin, Maimun Z Masafumi Matsuo Masdar Muid Masdar Muid Masruroh Rahayu Masruroh Rahayu, Masruroh Mayangsari, Elly Mentari Sekar Arum Miftahu Soleh Moch. Aris Widodo, Moch. Aris Mochammad Dalhar Mohammad Hidayat Mohammad Hidayat Mohammad S. Rohman Muhammad Ali Faisal Muhammad Rasjad Indra Muhammad Rasjad Indra Nabila Andjani, Nabila Nabila Erina Erwan Nashi Widodo Nathania Bella Claresta Niarti Ulan Sari Siarnu Nindy Sabrina Niniek Burhan Nisrina Nur Afina, Nisrina Nur Nunuk Helilusiatiningsih Nur Kusmiyati Nur Permatasari Nura Fattah Cantika Yoga Nurshalilah Nurshalilah Pande Made Dwi Jayasa Pande Made Dwijayasa Prima Wijayanti, Prima Primawardani, Putri Puryatni, Anik - Putri, Dianita Setya Pradita Putu Juniartha Raisa, Neila Raudhatul Jannah Restuningwiyani, Sintha Retnani, Diah Prabawati Retty Ratnawati Retty Ratnawati Rizky Amalia RR. Ettie Rukmigarsari Salim, Irfan Agus Samsul Arifin Santoso, Giovani Anggasta Satrijo, Budi Selvi Marcellia Septi Nur Rachmawati Setiyaningsih, Fera Yuli Setyawati Soeharto Setyawati Soeharto, Setyawati Shintiya Dewi, Debby Shofiyah, Fita SIMON BAMBANG WIDJANARKO Sintha Restuningwiyani Siti Lintang Kawuryan Siti Narsito Wulan Soetjahjo, Bintang Sri Andarini Sutrisno Sutrisno Syahrul Chilmi, Syahrul Syarifah Rohaya Tatit Nurseta Tatit Nurseta Tatit Nurseta Tika Ardhini Wardoyo Tinny Endang Tinny Endang Hernowati Tokunaga, Chiharu Tri Dewanti Widyaningsih Triawanti Triawanti Tuda, Josef Sem Berth Umar Zein Vierlia, Wino Vrieda Widowati, Hesty Widya Rahmawati Winda Nurtika Yonezawa, Kazuyoshi Yoshino, Ken-ichi Yuda Fibrianto Yudanto, Hendy Setyo Zuly Vita Aulya, Zuly Vita