Month: May 2020

Put together, these data implicate a major role of circulating leukocytes in influencing disease outcomes in influenza infection. We found that the systemic host response in severe infection differs significantly from that of mild infection. The main differences lay in the cell cycle and apoptosis pathways. Unexpectedly, immune response pathways did not differ significantly between infected groups. Other than TNF and IL-beta, inflammation-related genes that are well established in influenza infection do not discriminate between these groups. Also, interferon response genes do not differ significantly between mild and severe influenza infection. The lack of correlation among established immune/inflammatory markers led us to postulate that disease progression is determined by changes occurring elsewhere, such as in the cell cycle and apoptosis pathways. Further analyses revealed that there is a significantly greater number of cell cycle pathways activated in severe influenza infection compared to mild infection. In addition, the Severe group shows a greater up-regulation of genes encoding for key cell cycle proteins. These cell cycle proteins include cyclin and their associated catalytic kinase enzymes, namely, cyclin E, cyclin A, cyclin B, CDK1 and CDK2. Furthermore, this up-regulation is accompanied by an extensive activation of DNA replication machinery, including the pre-replication complex assembly, MCM complex and Cdt1. The heightened DNA replication activity does not seem to be host cell initiated because cyclin D, the initiator of cell cycle, is paradoxically down-regulated. Importantly, the increased DNA BAY 73-4506 structure synthesis occurs in the context of an abnormally low leukocyte response to infection, indicating that it is not a physiologically normal response. Despite an increase in DNA synthesis, paradoxical changes were seen in the mitotic phase. Here, we found up-regulation of genes opposing the completion of mitosis, including those encoding Securins and the Condensin Complex. Furthermore, there is strong activation of the spindle checkpoint complex, the cellular sensing system that normally prevents premature separation of chromosomes. Together, these proteins maintain chromosome condensation and their up-regulation is known to be associated with delayed mitotic exit. To understand the mechanism underlying this finding, we focused on the anaphase promoting complex, the major regulatory complex that coordinates cell cycle progression and exit from mitosis, which was also the most statistically significant pathway found in our analysis. Here we found abnormal changes in APC and its two co-activators. In subjects with a severe infection, CDC20 is unusually upregulated whilst no activation is seen in hCDH1. Most importantly, the APC gene is not expressed at all. In summary, severe influenza infection is characterized by opposing changes in cell cycle activity and these changes are associated with dysregulated cell cycle control.

The LMNA G608G transgenic mice targeted the expression of the Nutlin-3 Hutchinson-Gilford progeria syndrome mutation in keratin-5-expressing tissue led to a typical phenotype of HGPS. LMNA H222P mutated gene knockin mice exhibited conduction defects, chamber dilation, increased fibrosis and lack of hypertrophy, and also showed muscular dystrophy and death at 4–9 months of age. The patients with heterozygous for the LMNA E82K mutation showed clinical phenotypes of heart dilation and associated with conduction system disease at their onset age of 32 or 33 years. The two LmnaE82K transgenic mice lines exhibited chamber dilation, increased heart weights, increased fibrosis, upregulation of hypertrophic maker expression, nuclear structure defects and conduction defects, which was similar with the phenotypes of the patients carrying the LMNA E82K mutation. The importance of BNP as a diagnostic and therapeutic modality in cardiovascular disease is well known, it also acts as a local regulator of ventricular remodeling and a modifier of cardiac gene expression. Acta1 is present in the developing heart and it constitutes up to 20% of the striated actin of the adult heart. Since Acta1 is a multifunctional protein that interacts with many proteins involved in folding, polymerisation, contractility and regulation of contractility, abnormal levels may affect any of those functions. In the normal adult heart, approximately 2 to 4% of the myocardium is made up of collagen. The Col3a1 is one of the essential components of the cardiovascular extracellular matrix, maintaining structural and functional integrity of myocardium and thought to be responsible for abnormal myocardial stiffness and for the impaired pumping capacity of the heart. LMNA E82K mutation located in the coil 1B domain of central a helical rod domain of the lamin A and the lamin C proteins, those were conserved regions of the rod domain which have been shown to play crucial roles in the assembly of intermediate filament dimers into higher order oligomers. Mutations affect this region of IF proteins and may disrupt the interaction between the monomers and are linked to several diseases. Members of the intermediate filament superfamily are critical mechanical integrators of the nuclear membrane and the cytoskeleton, protecting the cell from repeated mechanical stress. Mutations in the lamin A/C gene may cause cardiomyopathy by weakening nuclei, which increase the fragility of nuclei and could be particularly harmful to muscle cells. Forces generated during muscle contraction might potentially lead to preferential breakage of nuclei containing a defective nuclear lamina.

The development of single or double molecule therapeutics for BoNT will require enhancement of the ability of mAbs to clear BoNT from the blood circulation. In this study, we tested the ability of a novel FP to enhance the neutralizing capacity of BoNT-specific mAbs at the level of the blood circulation. The FP is a bifunctional molecule that allows the adherence of biotin-conjugated immune complexes to the RBC surface. The FP was able to significantly increase the neutralization potency of BoNT-specific mAbs. When combined with the FP, the non-neutralizing 13A and 30B mAbs were able to fully neutralize lethal doses of BoNT/A and BoNT/B, respectively. The FP increased the quantity of BoNT/A that could be neutralized by the 6A mAb, the 4LCA mAb, and the 6A/4LCA mAb combination. These levels were achieved with much lower quantities of mAbs than we had previously required for protection with naked mAbs. The NIH and the FDA have not yet established a standard for protective efficacy for BoNT passive immune therapies, but we can compare the potency of our FP combinations with the approved BoNT anti-toxins. The human antiserum for use in infants, BabyBig, has a BoNT/A-specific potency of 3,000 LD50/mg. We found that the FP combined with 6A and 4LCA is sufficient to provide neutralization of 10 LD50 BoNT/A for 72 hours and partial protection at 96 hours. This result indicates the presence of physiologically relevant concentrations of the FP:mAb combination in vivo during this time. We also tested the FP:mAb combination in a post-exposure model, in which the FP:Ab was administered following a lethal dose of toxin. An intravenous injection of the combination was able to provide complete protection from a lethal intraperitoneal BoNT dose at 2 hours and partial protection at 4 hours. BoNT distribution experiments have demonstrated that the window of opportunity of exposure of a lethal dose of BoNT is determined by a first-order reaction that depends on the amount of BoNT administered and the period of time that elapses before the countermeasure is administered. Thus, anti-toxins that are sufficient to neutralize an entire circulating dose of BoNT should give the same window of opportunity for post-exposure salvage, and our post-exposure results are comparable to results LY2109761 obtained by others. The proposed mechanism through which the FP augments antibody neutralization activity involves binding of FP:mAb complexes to the RBC surface. Flow cytometry experiments in vitro demonstrated that the FP:mAb complexes are able to bind to erythrocytes and that the complexes serve as a molecular bridge to adhere BoNT to the RBC. Experiments with un-biotinylated antibodies and the FP did not show any enhancement of neutralization. Streptavidin alone did not improve neutralizing activity of biotinylated BoNT antibodies, in comparison to the RBC-targeted FP. Lastly, the levels of BoNT circulating in the plasma of mice that had received the FP in combination with the 4LCA and 6A were lowered.

Some studies have evaluated the volume status of PD patients in relation to modality transport status, residual renal function, or inflammation. However, whereas these studies contribute information on relative volume status in different groups of PD, they were hampered to express the degree of true fluid overload due to the lack of a reference population. In contrast, Wieskotten et al evaluated a large cohort of 688 healthy persons using the BCM to derive reference ranges, allowing to compare fluid overload as measured by BCM to age and gender matched values of the normal healthy population. In addition, expressing extracellular and intracellular water as absolute values induces the problem of scaling to body size. In previous studies using bio-impedance, ratios of extracellular water to height, weight, body surface area, intracellular water or total body water have been used to express “fluid overload”, but the ideal scaling parameter remains a matter of debate. The use of relative Dtissue hydrationdiminishes the problem of scaling nearly completely, and allows comparison to the healthy population. In HD patients relative Dtissue hydration is associated with mortality, indicating the clinical relevance of this parameter. The European Body Composition study in PD was designed to measure hydration status in a large, multicentric cohort of PD patients using the BCM device, as compared to a healthy reference population, and to establish associations between clinical and practice related parameters and volume status. The EuroBCM study is the first large multi-centre study of hydration status and its associated factors in PD patients in Europe allowing comparison to a healthy reference population. Fluid overload was a frequent finding in PD patients as compared to a healthy reference population, but comparable to that reported in HD patients. The deviation from the relation between blood pressure and tissue hydration was substantial, pointing out that blood pressure is not a good tool to evaluate hydration status in PD patients. Overhydration was associated with higher age, male gender, diabetes, lower BMI, higher systolic blood pressure, and use of hypertonic solutions, and in these conditions, physicians should have enhanced awareness for volume status. Use of LEE011 polyglucose or biocompatible glucose solutions or the type of PD modality was not independently associated with hydration status. In the large cohort of the EuroBCM in PD study, a substantial portion of patients were fluid overloaded by more than 1.1 litre, the 90th percentile of absolute Dtissue hydration in the normal reference population, and 25% of patients had a relativeD tissue hydration/extracellular water ratio above 15%, a value associated with increased mortality in HD patients. Substantial fluid overload is therefore indeed a prevalent problem in PD patients, and more attention should be given to its assessment and correction.

Furthermore, the CNV regions used in our study were greater than 1 Mb in length. There might also be important genes in smaller regions or in deleted regions, which were not immediately available from the original publication. However, the CNV regions we used in the current study were directly from the original publication and were of high quality. Nevertheless, our convergent strategy was able to identify 20 highpriority genes within strong biological context. Inclusion of more complete datasets or other types of datasets in future will greatly improve the quality of the current work. In conclusion, we proposed a stepwise enrichment procedure to converge CNV genes by incorporating publically available highthroughput datasets. By applying a subnetwork construction algorithm, we established a subnetwork for CNV genes, as well as for a set of previously reported genes for epilepsy. The overlap between the two subnetworks constitutes a high-priority candidate gene set for epilepsy. Integration of additional gene expression data further narrowed the candidate gene list to two of special interest. This procedure is extensible to other disorders. Euvolemia is a predictor of outcome in peritoneal dialysis patients, as volume overload is related to cardiac dysfunction, inflammation and mortality. Euvolemia is probably a more important adequacy parameter than small solute clearance, as fluid status but not small solute clearance predicts outcome. Guidance on how to achieve and maintain euvolemia in individual PD patients is hampered by the absence of a convenient device to measure volume status, and by the lack of insight in the prevalence of and factors associated with volume overload. In clinical practice, the assessment of volume status is relatively crude. Volume status is often assessed indirectly by measuring fluid removal, failing to take into account fluid balance by omission of dietary fluid intake. Ultrasonic evaluation of inferior vena cava diameter only assesses intravascular volume, and is also influenced by diastolic dysfunction , and is thus a reflection of preload, and not of tissue hydration. Parameters, such as Brain Natriuretic Peptide or NT-proBNP can reflect changes in hydration status, but are also influenced both by preload and ventricular abnormalities, and in patients with renal failure, accumulation can occur. Direct measurement of CHIR-99021 extracellular and total body water by dilution methods is considered as the golden standard, but these techniques are laborious and expensive. Bio-impedance spectroscopy represents a different approach to the assessment of fluid status. By measuring the flow of electrical current through the body, resistance and reactance can be measured, and in BIS, this is performed at different frequencies. The Body Composition Monitor is a bio-impedance spectroscopy device for clinical use, validated by isotope dilution methods and has been used in hemodialysis and PD.