Stiffer optical tweezers through real-time feedback control

Using real-time re-programmable signal processing we connect acousto-optic steering and back-focal-plane interferometric position detection in optical tweezers to create a fast feedback controlled instrument. When trapping 3 µm latex beads in water we find that proportional-gain position-clamping increases the effective lateral trap stiffness ~13-fold. A theoretical power spectrum for bead fluctuations during position-clamped trapping is derived and agrees with the experimental data. The loop delay, ~19 µs in our experiment, limits the maximum achievable effective trap stiffness

Technical Changes in Paraspinous Muscle Flap Surgery Have Increased Salvage Rates of Infected Spinal Wounds

Objectives: The objective of this study is to introduce modifications in paraspinous muscle flap surgery and compare this new variation's ability to salvage infected hardware with the classic technique. Infected posterior spine wounds are a difficult problem for reconstructive surgeons. As per experience, hardware retention in infected wounds maintains spinal stability, decreases length of stay, and decreases the wound healing complication rate. Methods: An 11-year retrospective office and hospital chart review was conducted between July 1996 and August 2007. All patients who underwent paraspinous muscle flap reconstruction for postspine surgery wound infections during this time period were included. There were 51 patients in the study representing the largest reported series, to date, for this procedure. Twenty-two patients underwent treatment using the modified technique and 29 patients were treated using the classic technique. Results: There was no statistical difference between the 2 groups in demographics, medical history, or reason for initial spine surgery. The hardware salvage rate associated with the modified technique was greater than the rate associated with the classic technique (95.4% vs 75.8%; P = .03). There were fewer postreconstruction wound healing complications requiring hospital readmission in the modified technique group than the classic group (13.6% vs 44.8%; P = .04). Patients in the modified technique group demonstrated a shorter mean length of stay than the patients in the classic group (23.7 days vs 29.7; P = .25). Conclusions: The modified paraspinous muscle flap technique is an excellent option for spinal wound reconstruction, preservation of spinal hardware, and local infection control

Detection of intermediates and kinetic control during assembly of bacteriophage P22 procapsid

Bacteriophage P22 serves as a model for the assembly and maturation of other icosahedral double-stranded DNA viruses. P22 coat and scaffolding proteins assemble in vitro into an icosahedral procapsid, which then expands during DNA packaging (maturation). Efficient in vitro assembly makes this system suitable for design and production of monodisperse spherical nanoparticles (diameter ≈50 nm). In this work we explore the possibility of controlling the outcome of assembly by scaffolding protein engineering. The scaffolding protein exists in monomer-dimer-tetramer equilibrium. We address the role of monomers and dimers in assembly by using three different scaffolding proteins with altered monomer-dimer equilibrium (weak dimer, covalent dimer, monomer). The progress and outcome of assembly was monitored by time-resolved X-ray scattering which allowed us to distinguish between closed shells and incomplete assembly intermediates. Binding of scaffolding monomer activates the coat protein for assembly. Excess dimeric scaffolding protein resulted in rapid nucleation and kinetic trapping yielding incomplete shells. Addition of monomeric wild type scaffold with excess coat protein completed these metastable shells. Thus, the monomeric scaffolding protein plays an essential role in the elongation phase by activating the coat and effectively lowering its critical concentration for assembly

Mechanochemical action of the dynamin protein

Dynamin is a ubiquitous GTPase that tubulates lipid bilayers and is implicated in many membrane severing processes in eukaryotic cells. Setting the grounds for a better understanding of this biological function, we develop a generalized hydrodynamics description of the conformational change of large dynamin-membrane tubes taking into account GTP consumption as a free energy source. On observable time scales, dissipation is dominated by an effective dynamin/membrane friction and the deformation field of the tube has a simple diffusive behavior, which could be tested experimentally. A more involved, semi-microscopic model yields complete predictions for the dynamics of the tube and possibly accounts for contradictory experimental results concerning its change of conformation as well as for plectonemic supercoiling.Comment: 17 pages, 4 figures; typos corrected, reference adde

Revealing the density of encoded functions in a viral RNA

Nikesh Patel, et al, ‘Revealing the density of encoded functions in a viral RNA’, Proceedings of the National Academy of Sciences of the United States of America (PNAS), Vol. 112 (7): 2227-2232, February 2015, doi: http:dx.doi.org/10. 1073/pnas.1420812112. This article is freely available online through the PNAS open access option.We present direct experimental evidence that assembly of a single-stranded RNA virus occurs via a packaging signal-mediated mechanism. We show that the sequences of coat protein recognition motifs within multiple, dispersed, putative RNA packaging signals, as well as their relative spacing within a genomic fragment, act collectively to influence the fidelity and yield of capsid self-assembly in vitro. These experiments confirm that the selective advantages for viral yield and encapsidation specificity, predicted from previous modeling of packaging signal-mediated assembly, are found in Nature. Regions of the genome that act as packaging signals also function in translational and transcriptional enhancement, as well as directly coding for the coat protein, highlighting the density of encoded functions within the viral RNA. Assembly and gene expression are therefore direct molecular competitors for different functional folds of the same RNA sequence. The strongest packaging signal in the test fragment, encodes a region of the coat protein that undergoes a conformational change upon contact with packaging signals. A similar phenomenon occurs in other RNA viruses for which packaging signals are known. These contacts hint at an even deeper density of encoded functions in viral RNA, which if confirmed, would have profound consequences for the evolution of this class of pathogensPeer reviewedFinal Published versio

Ventral Hernia Repairs: 10 year Single Institution Review at Thomas Jefferson University Hospital

Abstract Background Definitive repair of recurrent ventral hernias using abdominal wall reconstruction techniques is an essential tool in the armentarium for general and plastic surgeons. Ramirez 1 et al describes the “component separation” technique to mobilize the rectus-abdominus internal oblique and external oblique flap to correct the defect. The recurrence rate of incisional hernias increases to 20% after gastric bypass or extensive weight loss.2 The incidence of ventral hernias after failed recurrent hernia repair increases to 40%.3 It has been reported that utilizing the sliding myofascial flap repair technique, the recurrence rate was reduced to 8.5%.4 Materials and Methods This retrospective institutional study reviews 10 years of myofascial flap reconstruction 1996-2006 at TJUH. Several techniques and prosthetic materials (alloderm, permacol, vicryl, composix) were used in our institutional review by multiple surgeons in this time period. Our goal is to identify risk factors (i.e. smoking, diabetes, obesity, size of defect, peripheral vascular disease, enterocutaneous fistula, infection) that predict or categorize patients that are at increased risk for failure of primary repair, measure the complication rates (i.e. infection, recurrence, seroma, hematoma) and evaluate the techniques and long term effectiveness of several prosthetic materials. Results Three thousand twenty ventral hernia repairs were performed at TJUH between 1996 and 2006. Two thousand three hundred eighty three approximated the rectus abdominus primarily and of these 645 utilized a component separation technique. The recurrence rate for component separations was 18.5% and 83% for primary repairs. The average follow up was 5.49 years. Statistically significant risk factors (p\u3c0.05) for recurrence were obesity (BMI\u3e30 kg/m2), age\u3e65 years, male gender, preoperative infection and postoperative seroma. Conclusion Myofascial flaps are a safe, reliable therapy for recurrent ventral hernias that addresses the population of patients that have failed conventional primary closure and reduce the recurrence rates greater than 40 percent to 18.5 percent in the carefully selected patient population

Spillback Effects of Expansion When Product-Types and Firm-Types Differ

Contrary to perspectives that credit firms with only limited abilities to undertake significant change successfully, recent research has demonstrated that firms often improve their performance after undertaking major expansion to their operations. In this paper, we build on a study by Mitchell and Singh (1993) to test for differences in expansion effects, depending on whether the new goods substitute for old products and whether the firm is a generalist or specialist participant in the industry. The analysis helps us understand when a business can undertake major change successfully. The results have implications for ecological and other definitions of the core of a business and highlight the necessity for firms to undertake changes even at considerable risk to their existing operations.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/68398/2/10.1177_014920639502100105.pd

A Missense Mutation in a Highly Conserved Alternate Exon of Dynamin-1 Causes Epilepsy in Fitful Mice

Dynamin-1 (Dnm1) encodes a large multimeric GTPase necessary for activity-dependent membrane recycling in neurons, including synaptic vesicle endocytosis. Mice heterozygous for a novel spontaneous Dnm1 mutation—fitful—experience recurrent seizures, and homozygotes have more debilitating, often lethal seizures in addition to severe ataxia and neurosensory deficits. Fitful is a missense mutation in an exon that defines the DNM1a isoform, leaving intact the alternatively spliced exon that encodes DNM1b. The expression of the corresponding alternate transcripts is developmentally regulated, with DNM1b expression highest during early neuronal development and DNM1a expression increasing postnatally with synaptic maturation. Mutant DNM1a does not efficiently self-assemble into higher order complexes known to be necessary for proper dynamin function, and it also interferes with endocytic recycling in cell culture. In mice, the mutation results in defective synaptic transmission characterized by a slower recovery from depression after trains of stimulation. The DNM1a and DNM1b isoform pair is highly conserved in vertebrate evolution, whereas invertebrates have only one isoform. We speculate that the emergence of more specialized forms of DNM1 may be important in organisms with complex neuronal function