Inhibition of hepadnaviral replication by polyethylenimine-based intravenous delivery of antisense phosphodiester oligodeoxynucleotides to the liver

Antisense oligodeoxynucleotides (ODNs) appear as attractive anti-hepatitis B virus (HBV) agents. We investigated in vivo, in the duck HBV (DHBV) infection model, whether linear polyethylenimine (lPEI)-based intravenous delivery of the natural antisense phosphodiester ODNs (O-ODNs) can prevent their degradation and allow viral replication inhibition in the liver. DHBV-infected Pekin ducklings were injected with antisense O-ODNs covering the initiation codon of the DHBV large envelope protein, either in free form (O-ODN-AS2) or coupled to lPEI (lPEI/O-ODN-AS2). Following optimization of lPEI/O-ODN complex formulation, complete O-ODN condensation into a homogenous population of small (20–60 nm) spherical particles was achieved. Flow cytometry analysis showed that lPEI-mediated transfer allowed the intrahepatic delivery of lPEI/O-ODN-AS2 to increase three-fold as compared with the O-ODN-AS2. Following 9-day therapy the intrahepatic levels of both DHBV DNA and RNA were significantly decreased in the lPEI/O-ODN-AS2-treated group as compared with the O-ODN-AS2-treated, control lPEI/O-ODN-treated, and untreated controls. In addition, inhibition of intrahepatic viral replication by lPEI/O-ODN-AS2 was not associated with toxicity and was comparable with that induced by the phosphorothioate S-ODN-AS2 at a five-fold higher dose. Taken together, our results demonstrate that phosphodiester antisense lPEI/O-ODN complexes specifically inhibit hepadnaviral replication. Therefore we provide here the first in vivo evidence that intravenous treatment with antisense phosphodiester ODNs coupled to lPEI can selectively block a viral disease-causing gene in the liver

Multiomics modeling of the immunome, transcriptome, microbiome, proteome and metabolome adaptations during human pregnancy.

MotivationMultiple biological clocks govern a healthy pregnancy. These biological mechanisms produce immunologic, metabolomic, proteomic, genomic and microbiomic adaptations during the course of pregnancy. Modeling the chronology of these adaptations during full-term pregnancy provides the frameworks for future studies examining deviations implicated in pregnancy-related pathologies including preterm birth and preeclampsia.ResultsWe performed a multiomics analysis of 51 samples from 17 pregnant women, delivering at term. The datasets included measurements from the immunome, transcriptome, microbiome, proteome and metabolome of samples obtained simultaneously from the same patients. Multivariate predictive modeling using the Elastic Net (EN) algorithm was used to measure the ability of each dataset to predict gestational age. Using stacked generalization, these datasets were combined into a single model. This model not only significantly increased predictive power by combining all datasets, but also revealed novel interactions between different biological modalities. Future work includes expansion of the cohort to preterm-enriched populations and in vivo analysis of immune-modulating interventions based on the mechanisms identified.Availability and implementationDatasets and scripts for reproduction of results are available through: https://nalab.stanford.edu/multiomics-pregnancy/.Supplementary informationSupplementary data are available at Bioinformatics online

Ontwikkelingen in Transport (Volume en Herkomst) Ruwe Olie 2000-2003

RIKZ heeft geïnventariseerd hoeveel ruwe olie uit welke landen is getransporteerd naar de haven van Rotterdam in het jaar 2000. Doel van dit onderzoek was een goed beeld te krijgen van de mate waarin de verschillende ruwe olies getransporteerd worden. Dit is belangrijk omdat elke olie zich anders zal gedragen (\u91verweren\u92) wanneer de olie bij een calamiteit zou uitstromen. Een voorlopige eerste vergelijking levert het volgende op: \u95 Import van ruwe olie uit de drie belangrijkste exportlanden in 2000 (Saudi-Arabië, Noorwegen en Verenigd Koninkrijk) is redelijk constant \u95 Import vanuit Rusland en de Baltische Staten is verdubbeld \u95 Import uit andere Midden-Oostenlanden (met name Kuweit en Irak) is sterk afgenomen.CALPRE

Quality of life of women with breast cancer undergoing radiotherapy using the Functional Assessment of Chronic Illness Therapy-Fatigue questionnaire

Marta Muszalik,1 Małgorzata Kołucka-Pluta,2 Kornelia Kędziora-Kornatowska,1 Joanna Robaczewska1 1Department and Clinic of Geriatrics, Nicolaus Copernicus University, Collegium Medicum in Bydgoszcz, 2Centrum of Oncology in Bydgoszcz, Bydgoszcz, Poland Objective: Breast cancer is one of the most common cancers in women, particularly among older women. This illness along with its treatment has a great impact on a woman’s subjective opinion of her quality of life and functioning in everyday life. The aim of this research was to assess the quality of life in women undergoing radiotherapy for the treatment of breast cancer. Patients and methods: The research was carried out in 120 patients with breast cancer undergoing radiotherapy in the Oncological Center in Bydgoszcz, Poland. Among the 120 examined patients, there were 30 women aged between 20–50 years and the remaining were over 50 years of age, including 42 women over the age of 60. Demographic and clinical data were collected and the Functional Assessment of Chronic Illness Therapy-Fatigue questionnaire (version 4) was used to assess health-related quality of life (HRQOL) of the patients. Statistical analyses were conducted using Statistica, version 10.0. Results: Patients with breast cancer undergoing radiotherapy rated their quality of life with an average of 113.83 points. Older patients above 71 years of age also displayed significantly higher HRQOL (122.70 points). A lower level of fatigue was noticed among patients ≤50 years and ≥71 years of age. Education and marital status also had an important impact on HRQOL. Educated women with a good financial situation had a significantly higher HRQOL, compared to those with a lower education and in poor living conditions. Conclusion: HRQOL and state of fatigue in breast cancer patients treated with radiotherapy depended upon their age. Both were high among women aged 71 years and above, while younger patients (51–70 years of age) had slightly lower values. Results suggest that sociodemographic factors influence the conditions of life of women treated with radiotherapy for breast cancer in a significant way. Overall, patients tolerated this type of treatment well. Keywords: quality of life, breast cancer, elderly women, radiotherapy, HRQOL, FACIT-

Identification of Antigenic Regions of Duck Hepatitis B Virus Core Protein with Antibodies Elicited by DNA Immunization and Chronic Infection

The induction of humoral response in ducks by DNA-based immunization against duck hepatitis B virus (DHBV) core protein (DHBc) was investigated. In addition, the amino acid specificity of the induced response was compared by using peptide scanning to that elicited either by protein immunization or during chronic DHBV infection. Immunization of ducks with a plasmid expressing DHBc protein led to the induction of a long-lasting antibody response able to specifically recognize viral protein in chronically infected duck livers. Peptide scanning analysis of anti-DHBc response induced during chronic DHBV infection allowed us to identify six major antigenic regions (AR1 to AR6). The reactivity spectrum of duck sera elicited by protein immunization appeared narrower and was restricted to only four of these antigenic regions in spite of higher anti-DHBc antibody titers. Interestingly, anti-DHBc antibodies induced by DNA-based immunization recognized five of six antigenic regions, and the epitope pattern was broader and more closely related to that observed in chronic viral infections. To gain more insight into the location of antigenic regions, we built a three-dimensional (3-D) model of DHBc protein based on human and duck core sequence alignment data and the HBc 3-D crystal structure. The results suggest that two identified antigenic regions (AR2, amino acids [aa] (64)T-P(84), and AR5, aa (183)A-R(210)) are located at positions on the protein surface equivalent to those of the two HBc major epitopes. Moreover, we identified another antigenic region (AR3, aa (99)I-I(112)) that was recognized by all sera from chronically infected, DNA- or protein-immunized ducks within the large 45-aa insertion in DHBc protein, suggesting that this region, which lacks HBc, is externally exposed

Residues critical for duck hepatitis B virus neutralization are involved in host cell interaction.

To date, no detailed analysis of the neutralization properties of duck hepatitis B virus (DHBV) has been reported, and it is not clear whether any of the known neutralization epitopes correspond to the viral receptor binding site or to sequences involved in the cell entry pathway. We demonstrate here that antibodies directed against two overlapping peptides (amino acids 83 to 97 and 93 to 107), covering the sequences of most DHBV pre-S neutralizing epitopes, both inhibit virus binding to primary duck hepatocytes and neutralize virus infectivity. An extensive mutagenesis of the motif 88WTP90, which is the shortest sequence of the epitope recognized by the virus-neutralizing monoclonal antibody (MAb) 900 was performed in order to define the amino acids involved in these interactions. Single point mutations within this epitope affected neither virus replication nor infectivity but abolished virus neutralization by MAb 900 completely. Interestingly, mutants with two and three consecutive residue replacements (SIP and SIH) within this epitope retained replication competence but were no longer infectious. The loss of infectivity of SIH and SIP mutant particles was associated with significantly reduced binding to primary duck hepatocytes and could be rescued by trans complementation with wild-type pre-S protein. Taken together, these results indicate that each amino acid of the DHBV pre-S sequence 88WTP90 is critical for recognition by the neutralizing MAb 900 and that replacement of the first two or all three residues strongly reduces virus interaction with hepatocytes and abrogates infectivity. These data imply that the motif 88WTP90 contains key residues which are critical for interaction with both the neutralizing MAb and the host cell

Metagenomic analysis with strain-level resolution reveals fine-scale variation in the human pregnancy microbiome.

Recent studies suggest that the microbiome has an impact on gestational health and outcome. However, characterization of the pregnancy-associated microbiome has largely relied on 16S rRNA gene amplicon-based surveys. Here, we describe an assembly-driven, metagenomics-based, longitudinal study of the vaginal, gut, and oral microbiomes in 292 samples from 10 subjects sampled every three weeks throughout pregnancy. Nonhuman sequences in the amount of 1.53 Gb were assembled into scaffolds, and functional genes were predicted for gene- and pathway-based analyses. Vaginal assemblies were binned into 97 draft quality genomes. Redundancy analysis (RDA) of microbial community composition at all three body sites revealed gestational age to be a significant source of variation in patterns of gene abundance. In addition, health complications were associated with variation in community functional gene composition in the mouth and gut. The diversity of Lactobacillus iners-dominated communities in the vagina, unlike most other vaginal community types, significantly increased with gestational age. The genomes of co-occurring Gardnerella vaginalis strains with predicted distinct functions were recovered in samples from two subjects. In seven subjects, gut samples contained strains of the same Lactobacillus species that dominated the vaginal community of that same subject and not other Lactobacillus species; however, these within-host strains were divergent. CRISPR spacer analysis suggested shared phage and plasmid populations across body sites and individuals. This work underscores the dynamic behavior of the microbiome during pregnancy and suggests the potential importance of understanding the sources of this behavior for fetal development and gestational outcome