Macrophages Recognize Size and Shape of Their Targets

Recognition by macrophages is a key process in generating immune response against invading pathogens. Previous studies have focused on recognition of pathogens through surface receptors present on the macrophage's surface. Here, using polymeric particles of different geometries that represent the size and shape range of a variety of bacteria, the importance of target geometry in recognition was investigated. The studies reported here reveal that attachment of particles of different geometries to macrophages exhibits a strong dependence on size and shape. For all sizes and shapes studied, particles possessing the longest dimension in the range of 2–3 µm exhibited highest attachment. This also happens to be the size range of most commonly found bacteria in nature. The surface features of macrophages, in particular the membrane ruffles, might play an important role in this geometry-based target recognition by macrophages. These findings have significant implications in understanding the pathogenicity of bacteria and in designing drug delivery carriers

Virome Analysis of Transfusion Recipients Reveals a Novel Human Virus That Shares Genomic Features with Hepaciviruses and Pegiviruses

To investigate the transmission of novel infectious agents by blood transfusion, we studied changes in the virome composition of blood transfusion recipients pre- and posttransfusion. Using this approach, we detected and genetically characterized a novel human virus, human hepegivirus 1 (HHpgV-1), that shares features with hepatitis C virus (HCV) and human pegivirus (HPgV; formerly called GB virus C or hepatitis G virus). HCV and HPgV belong to the genera Hepacivirus and Pegivirus of the family Flaviviridae. HHpgV-1 was found in serum samples from two blood transfusion recipients and two hemophilia patients who had received plasma-derived clotting factor concentrates. In the former, the virus was detected only in the posttransfusion samples, indicating blood-borne transmission. Both hemophiliacs were persistently viremic over periods of at least 201 and 1,981 days. The 5′ untranslated region (UTR) of HHpgV-1 contained a type IV internal ribosome entry site (IRES), structurally similar to although highly divergent in sequence from that of HCV and other hepaciviruses. However, phylogenetic analysis of nonstructural genes (NS3 and NS5B) showed that HHpgV-1 forms a branch within the pegivirus clade distinct from HPgV and homologs infecting other mammalian species. In common with some pegivirus variants infecting rodents and bats, the HHpgV-1 genome encodes a short, highly basic protein upstream of E1, potentially possessing a core-like function in packaging RNA during assembly. Identification of this new human virus, HHpgV-1, expands our knowledge of the range of genome configurations of these viruses and may lead to a reevaluation of the original criteria by which the genera Hepacivirus and Pegivirus are defined

Rapid preparation of nuclei-depleted detergent-resistant membrane fractions suitable for proteomics analysis

<p>Abstract</p> <p>Background</p> <p>Cholesterol-rich membrane microdomains known as lipid rafts have been implicated in diverse physiologic processes including lipid transport and signal transduction. Lipid rafts were originally defined as detergent-resistant membranes (DRMs) due to their relative insolubility in cold non-ionic detergents. Recent findings suggest that, although DRMs are not equivalent to lipid rafts, the presence of a given protein within DRMs strongly suggests its potential for raft association in vivo. Therefore, isolation of DRMs represents a useful starting point for biochemical analysis of lipid rafts. The physicochemical properties of DRMs present unique challenges to analysis of their protein composition. Existing methods of isolating DRM-enriched fractions involve flotation of cell extracts in a sucrose density gradient, which, although successful, can be labor intensive, time consuming and results in dilute sucrose-containing fractions with limited utility for direct proteomic analysis. In addition, several studies describing the proteomic characterization of DRMs using this and other approaches have reported the presence of nuclear proteins in such fractions. It is unclear whether these results reflect trafficking of nuclear proteins to DRMs or whether they arise from nuclear contamination during isolation. To address these issues, we have modified a published differential detergent extraction method to enable rapid DRM isolation that minimizes nuclear contamination and yields fractions compatible with mass spectrometry.</p> <p>Results</p> <p>DRM-enriched fractions isolated using the conventional or modified extraction methods displayed comparable profiles of known DRM-associated proteins, including flotillins, GPI-anchored proteins and heterotrimeric G-protein subunits. Thus, the modified procedure yielded fractions consistent with those isolated by existing methods. However, we observed a marked reduction in the percentage of nuclear proteins identified in DRM fractions isolated with the modified method (15%) compared to DRMs isolated by conventional means (36%). Furthermore, of the 21 nuclear proteins identified exclusively in modified DRM fractions, 16 have been reported to exist in other subcellular sites, with evidence to suggest shuttling of these species between the nucleus and other organelles.</p> <p>Conclusion</p> <p>We describe a modified DRM isolation procedure that generates DRMs that are largely free of nuclear contamination and that is compatible with downstream proteomic analyses with minimal additional processing. Our findings also imply that identification of nuclear proteins in DRMs is likely to reflect legitimate movement of proteins between compartments, and is not a result of contamination during extraction.</p

Effects of preoperative walking ability and patient's surgical education on quality of life and functional outcomes after total knee arthroplasty

ABSTRACT OBJECTIVE: Prospectively analyze the effect of preoperative walking status and the patient's surgical education on functional outcomes and the three dimensions of quality of life (QoL) (pain, physical function, and mental health) after elective total knee arthroplasty (TKA). METHODS: A comparative analysis on the QoL and functional outcomes in patients who underwent total knee arthroplasty between January 2014 and June 2015. To compare effects of the patient's walking status and knowledge of the surgical procedure on QoL and functional outcomes following TKA by means of SF-36 questionnaire, CES D10, VAS, KSS, KSFS, WOMAC, as well as Friedmann and Wyman scores, 10MWT, and 30-second timed chair test, assessed before the operation and one, three, and six months after the operation. RESULTS: There were 168 knees in 154 patients: 46.75% men and 53.24% women. 52.38% of knees had grade-III OA and 40.47% of knees had grade-IV OA. Preoperatively, SF-36 PCS was 33.2 and MCS was 35.4. Mean KSS and KSFS in females was 37.3 (16.2) and 31.5 (13.8); in males it was 49.2 (18.4) and 42.5 (15.7), respectively. Mean WOMAC scores were 64.2 in females and 56.5 in males. Mean VAS and CES D10 scores were 8.8 and 8.2 in females, and 6.9 and 6.4 in males, respectively. Post operatively at the first, third, and sixth month, significant improvements in QoL and mean SF-36, CES D10, VAS, KSS, KSFS, WOMAC, and Friedmann and Wyman scores were observed, as well as in the 10MWT and 30 s timed chair test scores. Patients with better preoperative functional activity and satisfactory understanding of TKA presented a better functional performance and achieved a good quality life (p< 0.01). DISCUSSION: Surgeons educate TKA candidates regarding the surgical procedure, the nature of implants, and how the procedure would affect their lifestyle and what their expectations from TKA should be. These crucial considerations should boost their confidence, enhancing their involvement and cooperation in post-surgical rehabilitation, thereby improving their QoL, functional results, and post TKA experience. CONCLUSION: TKA candidates with good preoperative walking ability and understanding of knee arthroplasty have better QoL in early and late post-surgery periods. Patient's lifestyle and understanding significantly enhances the postoperative functional ability

Screening of Fe-Deficiency Tolerance in Okra (&lt;i&gt;Abelmoschus esculentus&lt;/i&gt; L.) Through Hydroponic Culture

Screening for Fe deficiency tolerance in okra (Abelmoschus esculentus) Bangladeshi genotypes (‘BARI-1’, ‘Local variety’, ‘Orca Onamica’, and ‘Prince’) were studied based on different morphological and physiological parameters. Number of leaves, shoot height and weight were significantly reduced in ‘Orca Onamica’ and ‘Prince’, whereas ‘BARI-1’ and ‘Local variety’ did not show prominent decrease in the aforesaid growth parameters under Fe deficiency. Again, ‘Orca Onamica’ and ‘Prince’ showed significantly decreased root length and root biomass under Fe deficiency. In contrast, these parameters were unchangeable in ‘BARI’-1 and ‘Local variety’ in Fe shortage compared to controls. Furthermore, Fe deficiency caused severe decrease in chlorophyll (a and b) and Fe concentrations in leaves of ‘Orca Onamica’ and ‘Prince’ grown on hydroponic culture. In contrast, chlorophyll (a and b) and Fe concentrations were not significantly decreased in ‘BARI-1’ and ‘Local variety’ due to Fe deficiency. Based on these findings, tolerance to Fe deficiency in these okra cultivars can be categorized as: tolerant (‘BARI-1’ and ‘Local’), and sensitive (‘Orca Onamica’ and ‘Prince’). The ranking can be applied in plant breeding program and may have great advantage over conventional methods. This study also demonstrates the effectiveness of hydroponic culture as an efficient method to screen Fe-efficient crop plants