PubMed Health⌕ Search

Biomedical subjects

Junwei Li

Publications and source records attributed to Junwei Li.

14 recordsLinked to original sources

Insights into Genome Ejection by a Therapeutic phiKMV-like Bacteriophage.

Ar-KM is a phiKMV-like therapeutic bacteriophage used in clinical candidate phage therapy cocktails to treat lung infections caused by P. aeruginosa. Here, we present an integrative structural atlas of Ar-KM proteins using cryo-EM, proteomics, and bioinformatics. From a single purified Ar-KM preparation, we identified three distinct populations: mature DNA-filled virions, open-nozzle particles with ejection proteins extending from the tail, and closed-nozzle empty particles. Near-atomic-resolution reconstructions of all three states enabled us to build atomic models for eleven structural proteins. The mature virion revealed the pre-ejection conformation of three ejection proteins, gp41, gp42, and gp43, homologous to coliphage T7's gp14, gp15, and gp16, respectively. Unlike T7, peptidoglycan hydrolase activity associated with the ejectosome resides in the gp15-like periplasmic tunnel protein gp42, whereas in T7 the lysozyme-like domain is located at the N-terminus of gp16, underscoring the structural plasticity and evolutionary mosaicity of ejection proteins. We further identified a short α-helical factor, gp34, present in eight copies at the mismatched interface between the portal barrel and gp41. Gp34 forms a cage within the nozzle, acting as a molecular wedge that stabilizes the open conformation and permits gp41 to assemble into a hexameric channel during ejection. Evolutionarily, gp34 appears to be an ortholog of the essential gene gp7.3 in phage T7 and is conserved across sequenced phiKMV-like phages. We propose that this protein functions as an ejection protein assembly factor, stabilizing the open nozzle during infection and allowing the coordinated exit of ejection proteins and their assembly into a DNA-ejectosome.

Pseudomonas phages↗

Covalent gluing and postgluing of Langmuir-Blodgett monolayers at hydrocarbon surfaces.

Langmuir-Blodgett (LB) monolayers of 5,11,17,23,29,35-hexaformyl-37,38,39,40,41,42-hexakis(1-n-octyloxy)calix[6]arene (2), deposited onto silylated silicon wafers, were cross-linked (i.e., "covalently glued") via Schiff base formation with poly(allylamine). Direct evidence for imine formation was obtained from X-ray photoelectron spectroscopy and from attenuated total reflection IR spectroscopy. These modified surfaces could be removed from the aqueous subphase into air with retention of the assembly and its orientation relative to the surface, as evidenced by atomic force microscopy, water contact angle measurements, and film thickness determinations by ellipsometry. Similar assemblies were also synthesized via a postgluing procedure, in which the substrate containing the LB monolayer was removed from the subphase and rapidly immersed into an aqueous solution containing poly(allylamine). The potential of combining postgluing methods with continuous LB film deposition as a surface modification technique is briefly discussed.

Journal Article↗

NoRC-dependent nucleosome positioning silences rRNA genes.

Previous studies have established that the Snf2h-containing chromatin remodeling complex NoRC mediates epigenetic silencing of a subset of rRNA genes (rDNA) by recruiting enzymatic activities that modify histones and methylate DNA. Here we have analyzed nucleosome positions at the murine rDNA promoter and show that active and silent rDNA copies are characterized not only by specific epigenetic marks but also by differently positioned nucleosomes. At active genes the promoter-bound nucleosome covers nucleotides from -157 to -2, whereas at silent genes the nucleosome is positioned 25 nucleotides further downstream. We provide evidence that NoRC is the molecular machine that shifts the promoter-bound nucleosome downstream of the transcription start site into a translational position that is unfavorable for transcription complex formation.

3T3-L1 Cells↗

Intergenic transcripts regulate the epigenetic state of rRNA genes.

Transcripts originating from the intergenic spacer (IGS) that separates rRNA genes (rDNA) have been known for two decades; their biological role, however, is largely unknown. Here we show that IGS transcripts are required for establishing and maintaining a specific heterochromatic configuration at the promoter of a subset of rDNA arrays. The mechanism of action appears to be mediated through the interaction of TIP5, the large subunit of the chromatin remodeling complex NoRC, with 150-300 nucleotide RNAs that are complementary in sequence to the rDNA promoter. Mutations that abrogate RNA binding of TIP5 impair the association of NoRC with rDNA and fail to promote H3K9&H4K20 methylation and HP1 recruitment. Knockdown of IGS transcripts abolishes the nucleolar localization of NoRC, decreases DNA methylation, and enhances rDNA transcription. The results reveal an important contribution of processed IGS transcripts in chromatin structure and epigenetic control of the rDNA locus.

Animals↗

Gluing Langmuir-Blodgett monolayers onto hydrocarbon surfaces.

This paper describes a new approach for modifying hydrophobic surfaces that is based on the use of ionically cross-linked (i.e., "glued") Langmuir-Blodgett monolayers. Specifically, this work shows how monolayers of 5,11,17,23,29,35-hexa(trimethylammonium methyl)-37,38,39,40,41,42-hexakis-n-hexadecyloxy-calix[6]arene (1a), which have been ionically cross-linked with poly(4-styrene-sulfonate) (PSS, MW 30,000-50,000), can be deposited onto silylated silicon wafers with their polar headgroups extending outward into air. This work also demonstrates the feasibility of using glued monolayers of 1a as "anchors" for attaching alternating layers of poly(diallyldimethylammonium) [PDADMA] and PSS ions onto these same hydrocarbon surfaces.

Journal Article↗

MR-visible coatings for endovascular device visualization.

PURPOSE: To investigate the potential utility of magnetic resonance (MR)-visible coatings for passive visualization of therapeutic endovascular devices such as catheters and guidewires. MATERIALS AND METHODS: Using a multistep coating process, gadolinium-based coatings were applied to commercially available off-the-shelf catheters and guidewires. These coated devices were imaged in phantoms made of fat-free yogurt, saline, and whole blood and also in live canine aorta on a 1.5-T cardiovascular MR scanner using T1-weighted two-dimensional radiofrequency (RF)-spoiled gradient-recalled echo, two-dimensional spin echo, and three-dimensional RF-spoiled gradient-recalled echo techniques. RESULTS: Commercially available off-the shelf catheters (4, 5, and 6 French) and guidewires (0.038 inch) were clearly visualized in all phantoms and canine aorta and the coatings proved to be durable and imageable without degradation in signal intensity up to 24 hours. MR-visible coatings address some of the shortcomings that have previously limited the role of MR as a guidance tool. CONCLUSION: Both in vitro and in vivo visualization of therapeutic endovascular devices coated with MR-visible coatings are found to be clinically viable.

4-Aminopyridine↗

Glued Langmuir-Blodgett bilayers having unusually high He/CO2 permeation selectivities.

Single Langmuir-Blodgett bilayers derived from 5,11,17,23,29,35-hexakis[(N,N,N-trimethylammonium)-N-methyl-37,38,39,40,41,42-hexakis-n-hexamedecyloxy-calix[6]arene hexachloride (1), which have been ionically cross-linked (i.e., "glued together") with poly(acrylic acid), have been found to exhibit He/CO2 permeation selectivities as high as 150. This degree of selectivity, for a membrane that is less than 6 nm in thickness, is without precedent. In principle, materials of this type could lead the way to improved membranes for hydrogen purification.

Journal Article↗

The chromatin remodeling complex NoRC controls replication timing of rRNA genes.

The ATP-dependent chromatin remodeling complex NoRC silences a fraction of mammalian ribosomal RNA genes (rDNA) by establishing heterochromatic structures at the rDNA promoter. Here we show that NoRC also plays a role in replication timing of rDNA. rDNA is replicated in a biphasic manner, active genes ( approximately 60%) replicating early and silent ones ( approximately 40%) replicating late in S-phase. The chromatin structure that marks active and silent rDNA repeats is propagated during cell division. To examine the function of NoRC in epigenetic inheritance and replication timing, we have monitored the chromatin structure, transcriptional activity and replication timing of rDNA in a cell line that moderately overexpresses NoRC. NoRC is exclusively associated with late-replicating rDNA arrays. Overexpression of NoRC silences rDNA transcription, reduces the size and number of nucleoli, impairs cell proliferation and resets replication timing from early to late. The results demonstrate that NoRC is an important determinant of replication timing and epigenetic marks are heritably maintained through DNA replication.

Adenosine Triphosphate↗

Exceptional gas permeation selectivity of a glued Langmuir-Blodgett bilayer by pH control.

The monolayer properties of 5,11,17,23,29,35-hexakis[(N,N,N-trimethylamonium)-N-methyl-37,38,39,40,41,42-hexakis-n-hexadecyloxy-calix[6]arene hexachloride (1) have been characterized over aqueous solutions of poly(acrylic acid) (PAA) and poly(methacrylic acid) (PMAA) as a function of pH. At high pH values (e.g., pH 10), such monolayers show relatively low surface viscosities. At low pH (e.g., pH 4.4), these monolayers exhibit relatively high surface viscosities. The barrier properties of single Langmuir-Blodgett bilayers of 1, which have been ionically cross-linked (i.e., "glued together") with PAA were found to correlate with changes in surface viscosity. Thus, bilayers that were fabricated under low pH conditions exhibited high permeation barriers and high permeation selectivity with respect to He and N(2). Given the extreme thinness of these glued bilayers (ca. 6 nm), the optimized He/N(2) selectivity of ca. 1000 is extraordinary. These results, taken together, demonstrate the feasibility of fine tuning the surface viscosity of monolayers of 1, and also the barrier properties of corresponding glued bilayers, by adjusting the pH of an aqueous subphase that contains a weak polyacid.

Acrylic Resins↗

Glued Langmuir-Blodgett bilayers from porous versus nonporous surfactants.

Poly(4-styrenesulfonate) (PSS) has been used to ionically cross-link (glue together) single Langmuir-Blodgett bilayers derived from 1,2,4,5-tetrakis[N,N-dimethyl-N-(1-hexadecyl)-ammoniummethyl]benzene (2a) and tris(N,N-dimethyl-N-hexadecylammoniummethyl)mesitylene (3). The resulting films are of high quality as judged by He/N2 permeation selectivities of ca. 100. Such selectivity is well in excess of the Knudsen diffusion limit of 2.6 and approaches that which has been found for glued bilayers of 5,11,17,23,29,35-hexakis[(N,N,N-trimethylamonium)-N-methyl-37,38,39,40,41,42-hexakis-n-hexamedecyloxy-calix[6]arene hexachloride (1). The significance of these findings to LB technology, in general, is briefly discussed.

Journal Article↗

The nucleolar remodeling complex NoRC mediates heterochromatin formation and silencing of ribosomal gene transcription.

Epigenetic control mechanisms silence about half of the ribosomal RNA (rRNA) genes in metabolically active cells. In exploring the mechanism by which the active or silent state of rRNA genes is inherited, we found that NoRC, a nucleolar remodeling complex containing Snf2h (also called Smarca5, SWI/SNF-related matrix-associated actin-dependent regulator of chromatin, subfamily a, member 5), represses rDNA transcription. NoRC mediates rDNA silencing by recruiting DNA methyltransferase and histone deacetylase activity to the rDNA promoter, thus establishing structural characteristics of heterochromatin such as DNA methylation, histone hypoacetylation and methylation of the Lys9 residue of histone H3. These results indicate that active and inactive rRNA genes can be demarcated by their associated proteins, and link chromatin remodeling to DNA methylation and specific histone modifications.

3T3 Cells↗

[Treatment of infected total knee arthroplasty].

OBJECTIVE: To investigate the treatment of infected total knee arthroplasty (TKA). METHODS: Between 1983 and 2000, 6 patients with infection after TKAs were treated, including 2 men and 4 women, aged on average 63 years (44 - 75 years). Initial knee arthroplasty was performed for osteoarthritis in 4 patients and for rheumatoid arthritis in 2 knees. The timing of diagnosis of infection after knee arthroplasty averaged 50 months (range, 1 month-11 years). Simple debridement and antibiotic treatment were prescribed for 3 patients, debridement and one-stage reimplantation for 1, debridement and two-stage reimplantation for 1, and athrodesis for 1. RESULTS: Of the 3 patients with simple debridement, one was cured, one failed but underwent athrodesis later, and one lost to follow up. Two patients with reimplantation were cured and had good function recovery. All of the 6 patients were followed up on average for 4 years. No infection recurred except one who lost to follow-up. CONCLUSIONS: Management of infection after total knee arthroplasty includes antibiotic suppression and debridement with prosthesis retention, insertion of another prosthesis as a one-stage or two-stage exchange technique, knee arthrodesis and amputation. These treatments have specific indications. To treat infection after total knee arthroplasty, suitable method should be taken according to patient's condition. Arthrodesis is the best salvage operation, though it may handicap patients' daily life. Reimplantation of another prosthesis could maintain a functional joint.

Adult↗

Wound infection after scoliosis surgery: an analysis of 15 cases.

OBJECTIVE: To discuss the causes and treatments of wound infections after scoliosis surgery. METHODS: Nine hundred and twenty-four caes of scoliosis were reviewed, and the clinical data of 15 cases of postoperative infection were analysed retrospectively. RESULTS: All 15 cases underwent spinal posterior fusion with autologous bone graft using instrumentations. Seven were diagnosed as early infection, and 8 were delayed infection. Radical debridement was performed in all 15 cases. The duration of antibiotics administration was 10 to 34 days with continuous closed irrigation for 2 to approximately 4 weeks and primary closure for the wounds. All patients were followed up for an average of 3.5 years (2 to 7.5 years) with good outcomes and no recurrence. CONCLUSION: Wound infection following surgical correction of scoliosis primarily results from intraoperative seeding, although host-related and operation-related factors may contribute to its development. Once the infections are diagnosed, good results can be achieved by prompt surgical debridement, irrigation and reasonably administered antibiotics. Removal of hardware may be necessary in deep infections.

Adolescent↗