cDNA micro array identification of a gene differentially expressed in adenovirus type 5- versus type 12-transformed cells

AbstractProteins encoded by non-oncogenic adenovirus type 5 and oncogenic adenovirus type 12 differentially affect expression of a number of cellular genes. We have used cDNA micro array analysis to identify a cellular gene that is expressed in Ad12- but not in Ad5-transformed cells. This cellular gene was found to be the gene encoding follistatin-related protein, a TGF-β inducible gene. Consistently, a constitutive factor binding to Smad binding elements was found in adenovirus type 12-transformed cells

Ekstrak Virgin Coconut Oil Sebagai Sumber Pangan Fungsional

Virgin Coconut Oil (VCO) merupakan salah satu jenis minyak nabati yang dapat bermanfaat dari aspek medis dan nutrisi karena dapat mencegah dan membantu mengobati penyakit tertentu serta dapat mempermudah proses pencernaan makanan dan penyerapan gizi. VCO dapat bertindak sebagai antioksidan dan antifotooksidan yang disebabkan oleh kandungan komponen minor (mikronutrien). Tujuan penelitian ini adalah mengekstrak komponen minor VCO menjadi produk untuk pangan fungsional yang mampu berperan sebagai antioksidan dan mengetahui peran VCO sebagai antiradikal bebas secara in vitro. Penelitian ini menggunakan metode eksperimental dengan 3 tahap yaitu: 1. Ekstraksi VCO menggunakan pelarut etanol dan metanol, 2. Pengujian penangkapan radikal bebas ekstrak VCO secara in vitro dengan metode 1,1-difenil-2-pikrilhidrazil (DPPH), 3. Identifikasi dan pengujian komponen kimia pada ekstrak VCO, identifikasi dan pengujian komponen minor pada ekstrak VCO dengan menggunakan High Performance Liquid Chromatography (HPLC). Hasil penelitian ini menunjukkan bahwa rendemen ekstrak VCO semakin tinggi dengan semakin tingginya persentase pelarut, ekstraksi dengan menggunakan etanol menghasilkan rendemen ekstrak yang lebih tinggi dibanding dengan menggunakan metanol. Aktivitas antioksidan dan kandungan total tokoferol dari ekstrak VCO semakin tinggi dengan semakin tingginya persentase pelarut yang digunakan. Salah satu senyawa tokoferol yang terdapat dalam VCO adalah -tokoferol. Berdasarkan analisa statistik menunjukkan bahwa etanol dan metanol menghasilkan sifat kimia ekstrak yang relatif sama, sehingga dapat direkomendasikan bahwa untuk mengekstraksi komponen minor dari VCO dapat menggunakan etanol karena di samping aman dari aspek kesehatan juga dapat menghasilkan rendemen ekstrak yang lebih tinggi

Implementation of Novel Molecular Biomarkers for Non-small Cell Lung Cancer in the Netherlands:How to Deal With Increasing Complexity

The diagnostic landscape of non-small cell lung cancer (NSCLC) is changing rapidly with the availability of novel treatments. Despite high-level healthcare in the Netherlands, not all patients with NSCLC are tested with the currently relevant predictive tumor markers that are necessary for optimal decision-making for today's available targeted or immunotherapy. An expert workshop on the molecular diagnosis of NSCLC involving pulmonary oncologists, clinical chemists, pathologists, and clinical scientists in molecular pathology was held in the Netherlands on December 10, 2018. The aims of the workshop were to facilitate cross-disciplinary discussions regarding standards of practice, and address recent developments and associated challenges that impact future practice. This paper presents a summary of the discussions and consensus opinions of the workshop participants on the initial challenges of harmonization of the detection and clinical use of predictive markers of NSCLC. A key theme identified was the need for broader and active participation of all stakeholders involved in molecular diagnostic services for NSCLC, including healthcare professionals across all disciplines, the hospitals and clinics involved in service delivery, healthcare insurers, and industry groups involved in diagnostic and treatment innovations. Such collaboration is essential to integrate different technologies into molecular diagnostics practice, to increase nationwide patient access to novel technologies, and to ensure consensus-preferred biomarkers are tested

A novel haemocytometric COVID-19 prognostic score developed and validated in an observational multicentre European hospital-based study

COVID-19 induces haemocytometric changes. Complete blood count changes, including new cell activation parameters, from 982 confirmed COVID-19 adult patients from 11 European hospitals were retrospectively analysed for distinctive patterns based on age, gender, clinical severity, symptom duration, and hospital days. The observed haemocytometric patterns formed the basis to develop a multi-haemocytometric-parameter prognostic score to predict, during the first three days after presentation, which patients will recover without ventilation or deteriorate within a two-week timeframe, needing intensive care or with fatal outcome. The prognostic score, with ROC curve AUC at baseline of 0.753 (95% CI 0.723-0.781) increasing to 0.875 (95% CI 0.806-0.926) on day 3, was superior to any individual parameter at distinguishing between clinical severity. Findings were confirmed in a validation cohort. Aim is that the score and haemocytometry results are simultaneously provided by analyser software, enabling wide applicability of the score as haemocytometry is commonly requested in COVID-19 patients

Protocollen en de praktijk, het blijft passen en meten

Editorial bij de bijdrage van A.J.A. Verhoeven, Y.M.A. van Maaren, B. Voermans en C. Botermans, getiteld ‘Prospectieve analyse van de bloedtransfusieketen met behulp van de SAFER-methode’

Cut-off values to rule out urinary tract infection should be gender-specific

The diagnosis of urinary tract infection (UTI) by urine culture is an expensive and time-consuming procedure. Using a screening method, to identify negative samples, would improve the procedure and reduce costs. In this study, urine flow cytometry, of over 7000 urine samples, was assessed by retrospective analysis. With a cut-off value of >200bacteria/μl, we obtained a sensitivity of 93.0%, a specificity of 63.5%, and a negative predictive value (NPV) of 96.2%. As a result the culturing of 49% of all samples could be avoided. In addition, the data was retrospectively analyzed to determine if the introduction of gender-specific cut-off values could improve screening results. The obtained receiver operator curves are indeed significantly different when gender specific cut-offs were used. When a NPV of 95% is considered acceptable the unisex cut-off value of >200bacteria/μl can be used for women (NPV 94.9%), but the cut-off value for men could be raised to >400bacteria/μl without diminishing the NPV (NPV 95.0%)

Sensitive detection and quantification of SARS-CoV-2 by multiplex droplet digital RT-PCR

The purpose of this study is to develop a one-step droplet digital RT-PCR (RT-ddPCR) multiplex assay that allows for sensitive quantification of SARS-CoV-2 RNA with respect to human-derived RNA and could be used for screening and monitoring of Covid-19 patients. A one-step RT-ddPCR multiplex assay was developed for simultaneous detection of SARS-CoV-2 E, RdRp and N viral RNA, and human Rpp30 DNA and GUSB mRNA, for internal nucleic acid (NA) extraction and RT-PCR control. Dilution series of viral RNA transcripts were prepared in water and total NA extract of Covid-19-negative patients. As reference assay, an E-GUSB duplex RT-PCR was used. GUSB mRNA detection was used to set validity criteria to assure viral RNA and RT-PCR assay quality and to enable quantification of SARS-CoV-2 RNA. In a background of at least 100 GUSB mRNA copies, 5 copies of viral RNA are reliably detectable and 10 copies viral RNA copies are reliably quantifiable. It was found that assay sensitivity of the RT-ddPCR was not affected by the total NA background while assay sensitivity of the gold standard RT-PCR assay is drastically decreased when SARS-CoV-2 copies were detected in a background of total NA extract compared with water. The present study describes a robust and sensitive one-step ddRT-PCR multiplex assay for reliable quantification of SARS-CoV-2 RNA. By determining the fractional abundance of viral RNA with respect to a human housekeeping gene, viral loads from different samples can be compared, what could be used to investigate the infectiveness and to monitor Covid-19 patients