Tingkat Pengetahuan Mahasiswa Fakultas Kedokteran Universitas Yarsi Mengenai Penggunaan Human Diploid Cell Dalam Proses Produksi Vaksin MR Dan Tinjauannya Menurut Islam

Latar Belakang : Vaksin MR merupakan salah satu vaksin yang wajib diberikan kepada anak 0 – 9 bulan. Produksi vaksin MR menggunakan Human Diploid Cell yang berasal dari janin yang sengaja di abortus menimbulkan kontroversi mengenai halal dan haram vaksin MR. Penggawa kesehatan masyarakat musti memahami dan memiliki dasar keilmuan untuk dapat menjawab kerisauan dan kontroversi mengenai kehalalan vaksin MR sehingga masyarakat menerima penggunaan vaksin MR sebagai bentuk preventif dari penyakit Measles. Menurut pandangan Islam, vaksin MR hukumnya mubah karena prinsip dharuriyah untuk menjaga hifdz nasb dan hifdz nafs. Metode : Jenis penelitian yang digunakan adalah penelitian deskriptif dengan pendekatan cross sectional menggunakan kuesioner. Populasi penelitian adalah mahasiswa fakultas kedokteran Universitas YARSI tahun pertama dan tahun ketiga yang memenuhi definisi operasional yang dipilih dengan teknik simple random sampling. Hasil : Penelitian yang dilaksanakan menggunakan kuesioner didapatkan dari 100 responden. Persentase jumlah kuesioner Pengetahuan mengenai Human Diploid Cell berdasarkan Tingkat Pendidikan didapatkan pengetahuan baik sebanyak 13,51% pada tahun ketiga dan 11,11% pada tahun pertama. Pengetahuan cukup sebanyak 62,16% pada tingkat ketiga dan 44,44% pada tahun pertama. Pengetahuan kurang sebanyak 24,32% pada tingkat ketiga dan 44,44% pada tahun pertama . Persentase jumlah kuesioner Pengetahuan mengenai Vaksin MR berdasarkan Tingkat Pendidikan didapatkan pengetahuan baik sebanyak 54,05% pada tahun ketiga dan 49,21% pada tahun pertama. Pengetahuan cukup sebanyak 43,24% pada tingkat ketiga dan 42,86% pada tahun pertama. Pengetahuan kurang sebanyak 2,70% pada tingkat ketiga dan 7,94% pada tahun pertama. Simpulan : Tidak terdapat hubungan antara Tingkat Pendidikan dengan pengetahuan mengenai Penggunaan Human Diploid Cell dalam Proses Produksi Vaksin MR. Menurut Islam, penggunaan vaksin MR menjadi mubah sebagai upaya menegakkan prinsip dharuriyah dalam menjaga keturunan bagi orang tua (hifdz nasb) dan menjaga nyawa anak dari ancaman penyakit Measles (hifdz nafs).

Tingkat Pengetahuan Mahasiswa Fakultas Kedokteran Universitas Yarsi Mengenai Penggunaan Human Diploid Cell Dalam Proses Produksi Vaksin Polio

Latar Belakang : Vaksin merupakan suspensi mikroorganisme yang dilemahkan atau dimatikan, atau protein antikgenik dari berbagai organisme tadi yang diberikan untuk mencegah, meringankan, atau mengobati penyakit-penyakit menular. Vaksin pertama kali tercatat pada tahun 1769, yang dipublikasikan oleh Edward Jenner, yaitu specimen yang berasal dari lesi lengan seseorang yang terinfeksi Cowpox. Human Diploid Cells (HDC) merupakan salah satu sel yang digunakan untuk mengkultur virus yang akan dijadikan vaksin. HDC yang berasal dari aborsi manusia ini banyak digunakan untuk mengkultur virus Polio IPV dan OPV, Rabies, Rubella, Measles, Varicella-Zooster, dan Hepatitis A. Tujuan : Vaksin polio merupakan vaksin yang diwajibkan pada anak yang dijadwalkan dari Ikatan Dokter Anak Indonesia (IDAI) yang dibagi menjadi dua jenis, IPV (Inactivated Polio Vaccine) dan OPV (Oral Polio Vaccine). Metode : Jenis Penelitian yang digunakan adalah deskriptif dengan pendekatan cross sectional menggunakan kuesioner. Populasi yang digunakan adalah mahasisa Fakultas Kedokteran Universitas YARSI tahun pertama dan tahun ketiga yang memenuhi syarat. Cara pemilihan sampel dengan simple random sampling. Hasil : Penelitian yang dilaksanakan selama 3 hari dengan menggunakan kuesioner, dari 100 responden didapatkan persentase jumlah kuesioner Pengetahuan mengenai Human Diploid Cell berdasarkan Tingkat Pendidikan didapatkan pengetahuan baik sebanyak 5% pada tahun ketiga dan 7% pada tahun pertama. Pengetahuan cukup sebanyak 23% pada tingkat ketiga dan 28% pada tahun pertama. Pengetahuan kurang sebanyak 9% pada tingkat ketiga dan 28% pada tahun pertama. Persentase jumlah kuesioner Pengetahuan mengenai Polio berdasarkan Tingkat Pendidikan didapatkan pengetahuan baik sebanyak 15% pada tahun ketiga dan 19% pada tahun pertama. Pengetahuan cukup sebanyak 18% pada tingkat ketiga dan 31% pada tahun pertama. Pengetahuan kurang sebanyak 4% pada tingkat ketiga dan 13% pada tahun pertama. Kesimpulan : Tidak terdapat hubungan antara tingkat pendidikan dengan pengetahuan mengenai Human Diploid Cell dalam vaksin Polio. Dalam pandangan Islam, penggunaan vaksin Polio hukumnya mubah karena prinsip Dharuriyat bertujuan untuk mempertahankan nyawa atau Hifdz an-nafs anak dari ancaman penyakit.

High concordance in preimplantation genetic testing for aneuploidy between automatic identification via Ion S5 and manual identification via Miseq

The Ion S5 (Thermo Fisher Scientific) and Miseq (Illumina) NGS systems are both widely used in the clinical laboratories conducting PGT-A. Each system employs discrepant library preparation steps, sequencing principles, and data processing algorithms. The automatic interpretation via Ion Reporter software (Thermo Fisher Scientific) and the manual interpretation via BlueFuse Multi software (Illumina) for chromosomal copy number variation (CNV) represent very different reporting approaches. Thus, it is intriguing to compare their ability of ploidy detection as PGT-A/NGS system. In the present study, four aneuploid cell lines were individually mixed with a diploid cell line at different aneuploid ratios of 0% (0:5), 10% (1:9), 20% (1:4), 40% (2:3), 50% (3:3), 60% (3:2), 80% (4:1) and 100% (5:0) to assess the sensitivity and specificity for whole chromosomal and segmental aneuploidy detection. The clinical biopsies of 107 blastocysts from 46 IVF/PGT-A cycles recruited between December 2019 and February 2020 were used to calculate the concordance. Initially, the pre-amplified products were divided into two aliquots for different library preparation procedures of each system. Applying the same calling criteria, automatic identification was achieved through the Ion Reporter, while well-trained technicians manually identified each sample through the BlueFuse Multi. The results displayed that both systems reliably distinguished chromosomal CNV of the mixtures with at least 10% aneuploidy from karyotypically normal samples ([Ion S5] whole-chromosomal duplication: 2.14 vs. 2.05, p value = 0.009, segmental deletion: 1.88 vs. 2.05, p value = 0.003; [Miseq] whole-chromosomal duplication: 2.12 vs. 2.03, p value = 0.047, segmental deletion: 1.82 vs. 2.03, p value = 0.002). The sensitivity and specificity were comparable between the Ion S5 and Miseq ([sensitivity] 93% vs. 90%, p = 0.78; [specificity] 100% vs. 100%, p value = 1.0). In the 107 clinical biopsies, three displayed chaotic patterns (2.8%), which could not be interpreted for the ploidy. The ploidy concordance was 99.04% (103/104) per embryo and 99.47% (2265/2277) per chromosome pair. Since their ability of detection were proven to be similar, the automatic identification in Ion S5 system presents comparatively faster and more standardized performance.

Sensitivity of Pipistrellus Nathusii Kidney Diploid Cell Strain to Viruses of Rhabdoviridae, Reoviridae, Bunyaviridae and Paramixoviridae Families

The discovery of a significant number of viral pathogens in bat tissue samples and excrement point to a potential prominent role of chiropters in the maintenance and spread of human and animal diseases. It also indicates the potential sensitivity of bat cells to a broad spectrum of viruses. The migratory pipistrelle bat, Pipistrellus nathusii, inhabits northeastern Europe and typically migrates to the southwest. Our study revealed the sensitivity (susceptibility) of a diploid cell line, derived from the kidney of P. nathusii to several transmissible animal disease causative agents such as Epizootic hemorrhagic disease, Akabane disease, Vesicular stomatitis virus and Peste des petit ruminants. High sensitivity of the P. nathusii kidney diploid cell line to viruses from various taxonomic groups allows them to be recommend for extensive use in virological studies.

Stromal and Tumor Glioma-Derived Cells with Similar Characteristics have Differences in α-Smooth Muscle Actin Expression and Localization

Gliomas are solid brain tumors composed of tumor cells and recruited heterogenic stromal component. The study of the interactions between the perivascular niche and its surrounding cells is of great value in unraveling mechanisms of drug resistance in malignant gliomas. In this study, we isolated the stromal diploid cell population from oligodendroglioma and a mixed population of tumor aneuploid and stromal diploid cells from astrocytoma specimens. The stromal cells expressed neural stem/progenitor and mesenchymal markers showing the same discordant phenotype that is typical for glioma cells. Moreover, some of the stromal cells expressed CD133. For the first time, we demonstrated that this type of stromal cells had the typical myofibroblastic phenotype as the α-SMA+ cells formed α-SMA fibers and exhibited the specific function to deposit extracellular matrix (ECM) proteins at least in vitro. Immunofluorescent analysis showed diffuse or focal α-SMA staining in the cytoplasm of the astrocytoma-derived, A172, T98G, and U251MG glioma cells. We could suggest that α-SMA may be one of the main molecules, bearing protective functions. Possible mechanisms and consequences of α-SMA disruptions in gliomas are discussed.

Evolution of dominance in gene expression pattern associated with phenotypic robustness

Background Mendelian inheritance is a fundamental law of genetics. When we consider two genomes in a diploid cell, a heterozygote’s phenotype is dominated by a particular homozygote according to the law of dominance. Classical Mendelian dominance is concerned with which proteins are dominant, and is usually based on simple genotype–phenotype relationship in which one gene regulates one phenotype. However, in reality, some interactions between genes can exist, resulting in deviations from Mendelian dominance. Whether and how Mendelian dominance is generalized to the phenotypes of gene expression determined by gene regulatory networks (GRNs) remains elusive. Results Here, by using the numerical evolution of diploid GRNs, we discuss whether the dominance of phenotype evolves beyond the classical Mendelian case of one-to-one genotype–phenotype relationship. We examine whether complex genotype–phenotype relationship can achieve Mendelian dominance at the expression level by a pair of haplotypes through the evolution of the GRN with interacting genes. This dominance is defined via a pair of haplotypes that differ from each other but have a common phenotype given by the expression of target genes. We numerically evolve the GRN model for a diploid case, in which two GRN matrices are added to give gene expression dynamics and simulate evolution with meiosis and recombination. Our results reveal that group Mendelian dominance evolves even under complex genotype–phenotype relationship. Calculating the degree of dominance shows that it increases through the evolution, correlating closely with the decrease in phenotypic fluctuations and the increase in robustness to initial noise. We also demonstrate that the dominance of gene expression patterns evolves concurrently. This evolution of group Mendelian dominance and pattern dominance is associated with phenotypic robustness against meiosis-induced genome mixing, whereas sexual recombination arising from the mixing of genomes from the parents further enhances dominance and robustness. Due to this dominance, the robustness to genetic differences increases, while optimal fitness is sustained to a significant difference between the two genomes. Conclusion Group Mendelian dominance and gene-expression pattern dominance are achieved associated with the increase in phenotypic robustness to noise.

High Concordance Between Automatic Identification in Ion S5 and Manual Identification in Miseq During Next-Generation Sequencing for Preimplantation Genetic Testing of Aneuploidy

The Ion S5 (Thermo Fisher Scientific) and Miseq (Illumina) NGS systems are both widely used in the clinical laboratories conducting PGT-A. Each system employs discrepant library preparation steps, sequencing principles, and data processing algorithms. The automatic interpretation via Ion Reporter software (Thermo Fisher Scientific) and the manual interpretation via BlueFuse Multi software (Illumina) for chromosomal copy number variation (CNV) represent very different reporting approaches. Thus, it is intriguing to compare their ability of ploidy detection as PGT-A/NGS system. In the present study, four aneuploid cell lines were individually mixed with a diploid cell line at different aneuploid ratios of 0% (0:5), 10% (1:9), 20% (1:4), 40% (2:3), 50% (3:3), 60% (3:2), 80% (4:1) and 100% (5:0) to assess the sensitivity and specificity for whole chromosomal and segmental aneuploidy detection. The clinical biopsies of 107 blastocysts from 46 IVF/PGT-A cycles recruited between December 2019 and February 2020 were used to calculate the concordance. Initially, the pre-amplified products were divided into two aliquots for different library preparation procedures of each system. Applying with the same calling criteria, automatic identification was achieved through the Ion Reporter, while well-trained technicians manually identified each sample through the BlueFuse Multi. The results displayed that both systems reliably distinguished chromosomal CNV of the mixtures with at least 10% aneuploidy from karyotypically normal samples ([Ion S5] whole-chromosomal duplication: 2.14 vs. 2.05, p-value=0.009, segmental deletion: 1.88 vs. 2.05, p-value=0.003; [Miseq] whole-chromosomal duplication: 2.12 vs. 2.03, p-value=0.047, segmental deletion: 1.82 vs. 2.03, p-value=0.002). The sensitivity and specificity were comparable between the Ion S5 and Miseq ([sensitivity] 93% vs. 90%, p=0.78; [specificity] 100% vs. 100%, p-value=1.0). In the 107 clinical biopsies, three displayed chaotic patterns (2.8%), which could not be interpreted for the ploidy. The ploidy concordance was 99.04% (103/104) per embryo and 99.47% (2265/2277) per chromosome pair. Since their ability of detection were proven to be similar, the automatic identification in Ion S5 system presents comparatively faster and more standardized performance.

Liver fibrosis-induced muscle atrophy is mediated by elevated levels of circulating TNFα

Liver cirrhosis is a critical health problem associated with several complications, including skeletal muscle atrophy, which adversely affects the clinical outcome of patients independent of their liver functions. However, the precise mechanism underlying liver cirrhosis-induced muscle atrophy has not been elucidated. Here we show that serum factor induced by liver fibrosis leads to skeletal muscle atrophy. Using bile duct ligation (BDL) model of liver injury, we induced liver fibrosis in mice and observed subsequent muscle atrophy and weakness. We developed culture system of human primary myotubes that enables an evaluation of the effects of soluble factors on muscle atrophy and found that serum from BDL mice contains atrophy-inducing factors. This atrophy-inducing effect of BDL mouse serum was mitigated upon inhibition of TNFα signalling but not inhibition of myostatin/activin signalling. The BDL mice exhibited significantly up-regulated serum levels of TNFα when compared with the control mice. Furthermore, the mRNA expression levels of Tnf were markedly up-regulated in the fibrotic liver but not in the skeletal muscles of BDL mice. The gene expression analysis of isolated nuclei revealed that Tnf is exclusively expressed in the non-fibrogenic diploid cell population of the fibrotic liver. These findings reveal the mechanism through which circulating TNFα produced in the damaged liver mediates skeletal muscle atrophy. Additionally, this study demonstrated the importance of inter-organ communication that underlies the pathogenesis of liver cirrhosis.

Determining sensitivity of novel animal-derived cell cultures to clinical isolates of human enterovirus Echovirus 11 and Coxsackievirus B5

The cell culture laboratory of Ekaterinburg Research Institute of Viral Infections established and characterized diploid cell cultures of fetal porcine larynx, lungs, kidneys and muscles for the first time. Sensitivity of the novel animal-derived cell cultures to the priority clinical isolates of human enterovirus, Coxsackievirus B5 and Echovirus 11, was evaluated. Experimental studies with a Coxsackievirus B5 (CB5–8100) strain detected high sensitivity to the virus in the diploid laryngeal and renal cells and absence of such sensitivity in the cells of the lung and muscle tissues. We also documented the total absence of sensitivity to Echovirus 11 (111/RD) in all the cell cultures under study. The results will be used for further studies of animal-derived cell cultures. Certification of the novel fetal porcine renal and laryngeal cells will open up new opportunities for a wide variety of their uses in virology for studying the etiology of enteroviral infections caused by Coxsackievirus B5.

Saccharomyces spores are born prepolarized to outgrow away from spore-spore connections and penetrate the ascus wall

1.AbstractHow non-spore haploid Saccharomyces cells choose sites of budding and polarize towards pheromone signals in order to mate has been a subject of intense study. Unlike non-spore haploids, sibling spores produced via meiosis and sporulation by a diploid cell are physically interconnected and encased in a sac derived from the old cell wall of the diploid, called the ascus. Non-spore haploids bud adjacent to previous sites of budding, relying on stable cortical landmarks laid down during prior divisions, but since spore membranes are made de novo it was assumed that, as is known for fission yeast, Saccharomyces spores break symmetry and polarize at random locations. Here we show that this assumption is incorrect: Saccharomyces cerevisiae spores are born prepolarized to outgrow, prior to budding or mating, away from interspore bridges. Consequently, when spores bud within an intact ascus, their buds locally penetrate the ascus wall, and when they mate, the resulting zygotes adopt a unique morphology reflective of re-polarization towards pheromone, which we dub the derrière. Long-lived cortical foci containing the septin Cdc10 mark polarity sites, but the canonical bud site selection program is dispensable for spore polarity, thus the origin and molecular composition of these landmarks remain unknown. These findings demand further investigation of previously overlooked mechanisms of polarity establishment and local cell wall digestion, and highlight how a key step in the Saccharomyces life cycle has been historically neglected.