Synthetic rabbit-human antibody conjugate as a control in immunoassays for immunoglobulin M specific to hepatitis E virus

<p>Abstract</p> <p>Background</p> <p>In assays for anti-hepatitis E virus (HEV) immunoglobulin M (IgM), large volumes of the patient's sera cannot be easily obtained for use as a positive control. In this study, we investigated an alternative chemical method in which rabbit anti-HEV IgG was conjugated with human IgM and was used as a positive control in the anti-HEV IgM assay. Rabbit anti-HEV IgG was isolated from immune sera by chromatography on protein A-Sepharose and was conjugated with human IgM by using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) as a crosslinker.</p> <p>Results</p> <p>The specific anti-HEV IgG antibody titer was 100,000 times that of the negative control, i.e., prebleed rabbit serum. The results of anti-HEV IgM enzyme-linked immunosobent assay showed that the antibody conjugate was similar to anti-HEV IgM antibodies produced in humans. The results of a stability experiment showed that the antibody conjugate was stable for use in external quality assessment or internal quality control trials.</p> <p>Conclusions</p> <p>We concluded that the chemically conjugated rabbit-human antibody could be used instead of the traditional serum control as a positive control in the anti-HEV IgM assay.</p

Development of a new duplex real-time polymerase chain reaction assay for hepatitis B viral DNA detection

<p>Abstract</p> <p>Background</p> <p>Quantification of hepatitis B virus (HBV) DNA can be used for diagnosing HBV infection and monitoring the effect of antiviral therapy. However, probably because of mismatches between the template and primer/probe, HBV DNA in some HBV infections could not be detected using currently available commercial assays with single primer/probe. By aligning the HBV sequences, we developed a duplex real-time polymerase chain reaction (PCR) assay using two sets of primers/probes and a specific armored DNA as internal control (IC).</p> <p>Results</p> <p>The limit of the duplex real-time PCR assay was 29.5 IU/ml, whereas the specificity was 100%. The within-run precision coefficient of variation (CV) ranged from 1.02% to 2.73%, while the between-run CV ranged from 0.83% to 1.25%. The optimal concentration of armored DNA IC in the HBV DNA duplex real-time PCR assay was 1 000 copies/ml. Data from 69 serum samples with HBV infection showed that the performance of the duplex real-time PCR assay was comparable to that of the COBAS Ampliprep/Cobas Taqman (CAP/CTM) HBV assay and was superior to those of the domestic commercial HBV assays.</p> <p>Conclusions</p> <p>The duplex real-time PCR assay is sufficiently sensitive, specific, accurate, reproducible and cost-effective for the detection of HBV DNA. It is suitable for high throughput screening and frequent HBV DNA level monitoring.</p

Autoantibodies against the Catalytic Domain of BRAF Are Not Specific Serum Markers for Rheumatoid Arthritis

BACKGROUND: Autoantibodies to the catalytic domain of v-raf murine sarcoma viral oncogene homologue B1 (BRAF) have been recently identified as a new family of autoantibodies involved in rheumatoid arthritis (RA). The objective of this study was to determine antibody responses to the catalytic domain of BRAF in RA and other autoimmune diseases. The association between RA-related clinical indices and these antibodies was also assessed. METHODOLOGY/PRINCIPAL FINDINGS: The presence of autoantibodies to the catalytic domain of BRAF (anti-BRAF) or to peptide P25 (amino acids 656-675 of the catalytic domain of BRAF; anti-P25) was determined in serum samples from patients with RA, primary Sjögren's syndrome (pSS), systemic lupus erythematosus (SLE), and healthy controls by using indirect enzyme-linked immunosorbent assays (ELISAs) based on the recombinant catalytic domain of BRAF or a synthesized peptide, respectively. Associations of anti-BRAF or anti-P25 with disease variables of RA patients were also evaluated. Our results show that the BRAF-specific antibodies anti-BRAF and anti-P25 are equally present in RA, pSS, and SLE patients. However, the erythrocyte sedimentation rate (ESR) used to detect inflammation was significantly different between patients with and without BRAF-specific antibodies. The anti-BRAF-positive patients were found to have prolonged disease, and active disease occurred more frequently in anti-P25-positive patients than in anti-P25-negative patients. A weak but significant correlation between anti-P25 levels and ESRs was observed (r = 0.319, p = 0.004). CONCLUSIONS/SIGNIFICANCE: The antibody response against the catalytic domain of BRAF is not specific for RA, but the higher titers of BRAF-specific antibodies may be associated with increased inflammation in RA

Supervisi Akademik oleh Kepala Sekolah dalam Meningkatkan Kompetensi Profesional Guru Pendidikan Agama Kristen Sdn 02 Bengkayang

: This study: 1) the effectiveness of the supervision of an academic supervisor in enhancing the professional competence of teachers Christian education at SDN 02 Bengkayang, 2) the constraints faced by the Principal in conducting academic supervision, and a solution to the academic supervision Principal can improve the professional competence of teachers of Religious Education Christians in SDN 02 Bengkayang. The informants are Principal, Christian Religious Education teacher, and fifth graders of SDN 02 Bengkayang Christians. Data were collected through interviews, observation and documentation. Technique authenticity of data using triangulation sources. Analyzed using an interactive model. The results showed that: 1) the academic supervision made Principal quite effective in improving teachers\u27 professional competence Christian education reflected in an increasing. 2) Barriers experienced Principal in academic supervision is the lack of time to conduct academic supervision. 3) The solution of these obstacles is the delegation of tasks to senior teachers to conduct academic supervision

MicroRNA-Mediated In Vitro and In Vivo Direct Reprogramming of Cardiac Fibroblasts to Cardiomyocytes

Repopulation of the injured heart with new, functional cardiomyocytes remains a daunting challenge for cardiac regenerative medicine. An ideal therapeutic approach would involve an effective method at achieving direct conversion of injured areas to functional tissue in situ

Skywork: A More Open Bilingual Foundation Model

In this technical report, we present Skywork-13B, a family of large language models (LLMs) trained on a corpus of over 3.2 trillion tokens drawn from both English and Chinese texts. This bilingual foundation model is the most extensively trained and openly published LLMs of comparable size to date. We introduce a two-stage training methodology using a segmented corpus, targeting general purpose training and then domain-specific enhancement training, respectively. We show that our model not only excels on popular benchmarks, but also achieves \emph{state of the art} performance in Chinese language modeling on diverse domains. Furthermore, we propose a novel leakage detection method, demonstrating that test data contamination is a pressing issue warranting further investigation by the LLM community. To spur future research, we release Skywork-13B along with checkpoints obtained during intermediate stages of the training process. We are also releasing part of our SkyPile corpus, a collection of over 150 billion tokens of web text, which is the largest high quality open Chinese pre-training corpus to date. We hope Skywork-13B and our open corpus will serve as a valuable open-source resource to democratize access to high-quality LLMs