Applying antibodies inside cells: Principles and recent advances in neurobiology, virology and oncology

To interfere with cell function, many scientists rely on methods that target DNA or RNA due to the ease with which they can be applied. Proteins are usually the final executors of function but are targeted only indirectly by these methods. Recent advances in targeted degradation of proteins based on proteolysis-targeting chimaeras (PROTACs), ubiquibodies, deGradFP (degrade Green Fluorescent Protein) and other approaches have demonstrated the potential of interfering directly at the protein level for research and therapy. Proteins can be targeted directly and very specifically by antibodies, but using antibodies inside cells has so far been considered to be challenging. However, it is possible to deliver antibodies or other proteins into the cytosol using standard laboratory equipment. Physical methods such as electroporation have been demonstrated to be efficient and validated thoroughly over time. The expression of intracellular antibodies (intrabodies) inside cells is another way to interfere with intracellular targets at the protein level. Methodological strategies to target the inside of cells with antibodies, including delivered antibodies and expressed antibodies, as well as applications in the research areas of neurobiology, viral infections and oncology, are reviewed here. Antibodies have already been used to interfere with a wide range of intracellular targets. Disease-related targets included proteins associated with neurodegenerative diseases such as Parkinson's disease (α-synuclein), Alzheimer's disease (amyloid-β) or Huntington's disease (mutant huntingtin [mHtt]). The applications of intrabodies in the context of viral infections include targeting proteins associated with HIV (e.g. HIV1-TAT, Rev, Vif, gp41, gp120, gp160) and different oncoviruses such as human papillomavirus (HPV), hepatitis B virus (HBV), hepatitis C virus (HCV) and Epstein-Barr virus, and they have been used to interfere with various targets related to different processes in cancer, including oncogenic pathways, proliferation, cell cycle, apoptosis, metastasis, angiogenesis or neo-antigens (e.g. p53, human epidermal growth factor receptor-2 [HER2], signal transducer and activator of transcription 3 [STAT3], RAS-related RHO-GTPase B (RHOB), cortactin, vascular endothelial growth factor receptor 2 [VEGFR2], Ras, Bcr-Abl). Interfering at the protein level allows questions to be addressed that may remain unanswered using alternative methods. This review addresses why direct targeting of proteins allows unique insights, what is currently feasible in vitro, and how this relates to potential therapeutic applications

DEVELOPING NOVEL APPROACHES TO IMPROVE RESPONSE TO T CELL BASED CANCER IMMUNOTHERAPY

Recently, T cell based immunotherapies have moved to the forefront of cancer immunotherapy with the success of Adoptive T cell therapy (ACT) and Immune checkpoint blockade.ACT, where patients are treated with tumour infiltrating T cells (TILs), conferred a clinical response rate of ~50%. Treatment with anti-CTLA4 and anti –PD1 therapy, conferred response rates of up to 50%, greatly improving the overall survival of patients with advanced melanoma amongst other cancer types. Despite the encouraging outcomes, there are relatively low response rates coupled with the delay of weeks to months before tumour shrinkage can be appreciated. Thus, understanding what tumour intrinsic pathways contribute to non-responsiveness to immunotherapies and their effect on T cells, to improve response rates, shorten time to treatment effect and developing predictive biomarkers of response are vital to the care of cancer patients. In order to identify possible tumour intrinsic pathways that could be perturbed to improve responses to immunotherapy, a high-throughput in vitro screen with 850 different bio-active compounds (Selleckchem), was designed to search for agents that could either increase or decrease the resistance of melanoma tumour cells to T cell mediated killing. Paired tumour samples and TILs from melanoma patients were used to assess which compounds when used to treat the melanoma cell lines can enhance the cytotoxic activity of the TILs against the paired melanoma sample, using a flow cytometry based assay in which active caspase 3 was used as a read out of apoptosis. Heat shock protein 90 (HSP90) inhibitors amongst compounds that improved T cell mediated cytotoxicity. We show that treatment with the HSP90 inhibitor ganetespib (Synta) greatly improves T cell mediated cytotoxicity of both human and murine cancer cells lines in vitro. Furthermore, in vivo murine studies using the MC38/gp100 tumour model show that ganestespib in combination with immune checkpoint blockade, resulted in superior antitumour effect and survival compared to either treatment alone. Microarray analysis of human cell lines treated with ganetespib in vitro revealed an increase in interferon alpha (IFN-α) response genes including IFIT1, IFIT2 and IFIT3. Silencing IFIT genes abrogated the synergy observed with ganetespib treatment and T cell mediated killing, suggesting that the IFN-α response pathway plays an important role in this combination therapy. This work highlights the importance of IFIT genes in response to T cell based immunotherapy and will enable the emergence of a new combination therapy of HSP90 inhibitors and anti-CTLA4 for the treatment of melanoma patients that will increase the percentage of patients responding to immunotherapy and achieving long term responses. To understand what factors influence CD8 T cell effector function at the tumour site, we used a murine adoptive cellular therapy model in which B16, a gp100-expressing tumour cell line was implanted in mice and treated with transgenic Pmel T cells, which recognize gp100 in the context of H-2Db.Transferred Pmel T cells were recovered from the spleen and tumour of the mice. To interrogate novel pathways which may inhibit the functions of tumour-reactive T cells at the tumour site, microarray and genome-wide gene expression analyses, were used to characterize the differential expression profiles among Pmel T cells from different groups. 720 genes were differentially expressed by T cells recovered from the tumour site, when compared with those recovered from the spleen. Amongst them, was the transcription factors Runx2, a gene whose role has not been described in T cells, to be one of the genes that were differentially expressed. In addition, we found that when Runx2 was absent in T cells they did a better job at controlling tumour growth and vice versa. Further investigation revealed that Runx2 regulated processes in T cells such as cytokine production and differentiation status, processes which are important in controlling tumour growth. We are currently focused on understanding how Runx2 controls these processes in an effort to provide ways to improve existing immunotherapies or discover new ones

Pemanfaatan Tumbuhan Hutan sebagai Bahan Baku Unit USAha Jamu dan Pengolahan Bahan Alam di Kalimantan Selatan

The existence of forest plants have been the source of traditional medicine for local communities. The use of natural materials by local communitiies as part of the medicinal treatment is done for subsistence. Not many identifiable utilization of the forest plants to meet the needs of business units herbal raw materials and processing of natural materials. This study seeks to identify the utilization of forest plants as raw material for herbal medicine business unit and the processing of natural materials in South Kalimantan. Mapping the distribution of the manufacturing herbal and natural ingredients also conducted and analyzed its association with the presence of the plant and within the forest. Data is collected using semi-structured interview to further analyzed descriptively. The collection of layers that constitute thematic information for the analysis of Geographic Information Systems. GIS analysis using ArcView 3.3 software. The results showed that the mixture dipterocarp forest is a forest type that is the source material for herbal medicine business unit and the processing of natural materials in South Kalimantan. The business unit is the most widely used natural materials from the forest is the micro and small (≤ 50%) or medium sized business units are relatively few industries using natural materials from the forest (± 10%). The distribution of herbal medicine business unit and the processing of natural materials in South Kalimantan uneven. Herbal medicine business unit and the processing of natural materials is also limited to the city of Banjarmasin, Martapura, Banjarbaru, Amuntai, Barabai. Distance to the forest as a source of raw materials and distribution of herbal industry/processing of natural materials not correlate significantly. The rapid development of urban economy and cultural factors related to the presence of viscous processing units herbal and natural ingredients

Evaluasi Amdal Pembangunan Gedung 10 Lantai Di Pusat Kota Bandung

Pembangunan yang tidak terkendali di Kota Bandung berdampak pada tingginya tingkat pencemaran lingkungan. Hal ini mengisyaratkan bahwa AMDAL belum dapat dijadikan sebagai salah satu instrument untuk mitigasi bencana serta pelengkap bagi kelayakan lingkungan.Tujuan penelitian: (1) mengevaluasi dokumen kerangka acuan pembangunan gedung 10 lantai di pusat Kota Bandung, (2) mengevaluasi hasil analisis dampak lingkungan (ANDAL), (3) mengevaluasi RKL dan RPL, (4) mengevaluasi implementasi AMDAL pembangunan gedung 10 lantai di pusat Kota Bandung ditinjau dari ketepatan waktu, ketepatan tindakan dan ketepatan hasil guna mengurangi dampak negatif serta meningkatkan dampak positif. Metode pemecahan masalah dengan deskriptif dan teknik analisis analisis isi (content analysis). Hasil penelitian:1) dokumen KA disusun oleh konsultan dengan jumlah tenaga akhli kurang dari yang disyaratkan sehingga konsultan tidak mampu menelaah secara cermat, holistik dan integral dalam menyusun dokumen ANDAL, (2) studi ANDAL cenderung kekurangan informasi komponen- lngkungan yang berdampak negatif dan positif terhadap lingkungan, (3) para konsultan penyusun RKL- RPL tidak mengetahui secara tepat komitmen dan mekanisme koordinasi yang mengikat secara kelembagaan dalam merencanakan pengelolaan lingkungan untuk mengurangi dampak negatif serta meningkatkan dampak positif. (4) produk AMDAL hanya menjadi dokumen yang disimpan tanpa sangsi jika indikator-indikator ANDAL, RKL dan RPL dilanggar

Pengaruh Dominan Cash Ratio dan Debt To Asset Ratio terhadap Return On Equity dan Return On Assets pada Perusahaan Transportasi di Bursa Efek Indonesia

The Purpose of this research is to be achieved is to determine the most dominant influence between casht ratio and debt to asets ratio toward return on equity and return on Assets to transportation companies on the stock exchange Indonesia.In the research to provide information will be the dominant influence is cash ratio and debt to assets ratio to return on assets.This is because the cash ratio and debt to assets ratio and return on assets indicate how much contribution the companys assets

Post-Sekulerisme Islam Populis di Indonesia

This article explores the phenomenology of post-secularism in Indonesia. Populist Islamic movement strike for islamization public sphere as a sign of post-secularism in Indonesia. The islamization proceeded both in government dan the public sphere. These phenomena show that the community of urban Muslims can’t leave religious aspects in the public sphere. This research uses the qualitative-description method and library research models. The first result of this research shows that Islamic populism is coming from the urban Muslim middle class who have access to the modern world. Second, the populist Islamic movement who did islamization of the public sphere shows the strengthening of religion's role in the public sphere. 

Willingness to Pay and Ability to Pay for Health Care Services at Zainoel Abidin Public Hospital Banda Aceh, Indonesia

Increasing accessibility to health care services is a prime goal in many developing countries, purposively to create an equality health care services to the populations. However, there are peoples who still do not well-utilized the provided health care provision especially due to the financial reason. Willingness-to-pay (WTP) and Ability-to-pay (ATP) are two powerful tools to measure the health care services utilization in this study, as it would present the reflection of Acehnese purchasing power for health care provision in Aceh. The purpose of this study is to determine the risk factor for healt care utilization amongst outpatient at public hospital dr.Zainoel abidin Banda Aceh. Methods: cross-sectional study involved a total of 180 outpatient captured at polyclinic public hospital dr.Zainoel abidin Banda Aceh. Survey was carried out by using a tailor-made questionnaire comprised of four separated sections; patient\u27s sociodemography, perception, healthcare financing and insurance coverage. Data was statistically analysed by using SPSS software and had gone trough univariate analysis and multiple logistic regression. Majority respondents (83.9%) use public health care services. Their WTP as outpatient at public health care services majority in the range less than Rp.9000 while as inpatient they were willing to pay within the range of less than Rp.30.000. However, majority (71.1%) patients expressed their ability to pay for health care services were between Rp.15.000 to Rp.219.999. Most of the respondent do not have health insurance (64.4%). However, respondent that only able to pay less than Rp.15000 and between Rp. Rp.15000 to Rp. 219.999 for health care services have a preference of going to public health care provision rather than private to seek for health care services that is vice versa to those patients that have health insurance coverage both, patient\u27s ATP and possesion of health insurance coverage presented a significant different (p<0.05) with the public health care utilization in this study. The public health care utilization among Acehnese is closely influenced by two factors; their abillity to pay for health care services and their possession of health care insuranc

GIS Untuk Integrasi Interpretasi Substrat Dasar Perairan menggunakan Penggolahan Citra ALOS-AVNIR dan Side Scan Sonar

Interpretasi substrat dasar perairan merupakan salah satu kajian penting dalam ilmu kelautan, sebagai kajian utama maupun kajian pendamping untuk kajian fenomena kelautan. Substrat dasar perairan penting untuk diketahui karena sebarannya yang sangat dinamis, di kedalaman perairan yang sama bisa saja terdapat materi substrat yang berbeda dan materi substrat yang sama bisa terdapat di rentang kedalaman yang berbeda. Interpretasi memanfaatkan citra Side Scan Sonar  dan citra satelit ALOS AVNIR-2untuk sebagian perairan Selat Sunda. Metode yang digunakan adalah pengolahan citra (image processing) identifikasi substrat perairan untuk citra Side Scan Sonar dengan Sonarwiz dan penerapan algoritma Lyzenga dan transformasi NDVI (Normalize Different Vegetation Index) untuk data citra satelit. Transformasi NDVI meningkatkan akurasi pemetaan substrat pada citra ALOS AVNIR-2. Hasil pengolahan kedua citra dioverlay menggunakan GIS untuk menampilkan visualisasi sebaran substrat perairan.Kata Kunci :ALOS-AVNIR, Side Scan Sonar, GIS, Lyzeng

Kajian Potensi Energi Arus Laut Di Perairan Selat Antara Pulau Kandang Balak Dan Pulau Kandang Lunik, Selat Sunda

Energilistrikterbarukandari laut merupakan salah satu upaya dalam pencarian energi ditengah krisis bahan bakar minyak. Energi arus laut banyak terdapat di selat sebab pada selat tersebut terjadi penyempitan ruang gerak arus sehingga kecepatan arus menjadi lebih cepat. Salah satunya yaitu Selat Sunda di antara Pulau Kandang Balak dan Pulau Kandang Lunik. Tujuan penelitian adalah mengetahui potensi energi listrik yang berasal dari laut berdasarkan nilai kecepatannya di antara Pulau Kandang Balak dan Pulau Kandang Lunik, Selat Sunda.Pengukuran arus laut dengan metode Lagrangian di berbagai titik lokasi penelitian kemudian dilakukan analisa pendekatan model untukmeramalkankarakteristiknya. Hasilmenunjukkanbahwakecepatanarustertendah 0,008 m/s saatsurutdenganarah menuju Bakauheni sedangkan saat menuju surut memiliki kecepatan arus tertinggi yaitu 1,1 m/s dengan arah menuju Samudera Hindia. KecepatanarustertinggiberadapadajarakterdekatantaraPulauKandangBalakdanPulauKandangLunikyaitusekitar 100 meter