Intramedullary Spinal Epidermoid Cyst-A Rare Cause of Spastic Paraparesis.

Spinal intramedullary epidermoids are rare intramedullary lesions of the spinal cord. They may be congenital or acquired with the congenital type often associated with spinal dysraphism and other spinal anomalies. The clinical presentation depends on the level of the involvement of the spinal cord. Management of these lesions is surgical excision. We report a case of intramedullary spinal epidermoid who presented with spastic paraparesis.

Post-partum paraplegia following spinal anaesthesia: a report of two rare cases.

Pregnancy and lumbar puncture are rare instances that can precipitate sudden onset paraplegia in patients with otherwise slow-growing intradural tumours. Surgeons and anaesthesiologists should be aware of the etiological factors leading to pregnancy- and delivery-related rapid tumour growth and its complications. Lumbar puncture-related complications leading to acute precipitation of neurological symptoms must be addressed promptly for favourable outcome in such patients. We describe the report of two patients who developed acute onset paraparesis after spinal anaesthesia for caesarean section. Both were found to be having undiagnosed spinal tumours and managed surgically. We recommend urgent MRI in cases of acute onset non-resolving paraparesis in the peripartum period, for timely diagnosis and management of this rare clinical entity.

Differential diagnosis of BPOP arising in relation to patella.

INTRODUCTION: Solitary exostosis is common at the metaphysis of long bones, and rarely may it develop in the lower pole of the patella. Usually it stops growing after skeletal maturity unless complicated. When the growth continues after skeletal maturity, other rare possibilities need to be considered such as bizarre parosteal osteochondromatous proliferation (BPOP). Though solitary exostosis is common at the metaphysis of long bones, very rarely it also develops in lower pole of the patella. Usually they stop growing after skeletal maturity unless complicated. When it starts after skeletal maturity and continues to grow, other rare possibilities like bizarre parostealosteochondromatousproliferation (BPOP) are to be thought of. CASE REPORT: 21 years male student presented with anterior midline painless progressive swelling over right knee joint of one year duration which was hard, non-tender, fixed to patella but mobile with patella. X ray showed midline heterogeneously radio-opaque swelling attached to inferolateral aspect of the anterior surface of patella. Patellar out line is fully maintained except the narrow site of tumour attachment. After exposing through midline incision, the swelling was found to incorporate the patellar tendon completely and an anterior vertical midline cleavage was found. The mass was deliberately detached along the cleavage and from intact patellar tendon. Almost full range of knee movement is obtained in operation table. Immediate post operative 10° quadriceps lag was corrected with quadriceps setting exercises in two weeks time. Histopathological examination demonstrated thin layer of cartilage cover, irregular lamellar bone in deeper zone and spindle cells between them without cytoplasmic atypia. Plenty of cartilage cells in different stages of maturation are seen without column formation. Marrow elements are absent. Periosteum could not be demonstrated and there was no other evidence of malignancy. Features simulate 'bizarre parosteal osteochondromatous proliferation'. There is no recurrence in five years of follow up. CONCLUSION: When exostosis like lesions arise from unusual site and at an unusual age group, other rare conditions need to be investigated. Though the final diagnosis of BPOP is obtained after careful histo-pathological examination, the clinico-radiological findings are also relevant. As literature search indicates, this is possibly second incidence where BPOP arised from sesamoid bone and first from patella.

Antimicrobial functionalization of silicone surfaces with engineered short peptides having broad spectrum antimicrobial and salt-resistant properties.

Catheter-associated urinary tract infections (CAUTIs) are often preceded by pathogen colonization on catheter surfaces and are a major health threat facing hospitals worldwide. Antimicrobial peptides (AMPs) are a class of new antibiotics that hold promise in curbing CAUTIs caused by antibiotic-resistant pathogens. This study aims to systematically evaluate the feasibility of immobilizing two newly engineered arginine/lysine/tryptophan-rich AMPs with broad antimicrobial spectra and salt-tolerant properties on silicone surfaces to address CAUTIs. The peptides were successfully immobilized on polydimethylsiloxane and urinary catheter surfaces via an allyl glycidyl ether (AGE) polymer brush interlayer, as confirmed by X-ray photoelectron spectroscopy and water contact angle analyses. The peptide-coated silicone surfaces exhibited excellent microbial killing activity towards bacteria and fungi in urine and in phosphate-buffered saline. Although both the soluble and immobilized peptides demonstrated membrane disruption capabilities, the latter showed a slower rate of kill, presumably due to reduced diffusivity and flexibility resulting from conjugation to the polymer brush. The synergistic effects of the AGE polymer brush and AMPs prevented biofilm formation by repelling cell adhesion. The peptide-coated surface showed no toxicity towards smooth muscle cells. The findings of this study clearly indicate the potential for the development of AMP-based coating platforms to prevent CAUTIs.

Site specific immobilization of a potent antimicrobial peptide onto silicone catheters: evaluation against urinary tract infection pathogens.

Bacterial colonization of urinary catheters is a common problem leading to Catheter Associated Urinary Tract Infections (CAUTIs) in patients, which result in high treatment costs and associated complications. Due to the advantages of antimicrobial peptides (AMPs) compared to most other antimicrobial molecules, an increasing number of AMP-coated surfaces is being developed but their efficacy is hindered by suboptimal coating methods and loss of peptide activity upon surface tethering. This study aims to address this issue by employing a methodic approach that combines a simple selective chemical immobilization platform developed on a silicone catheter with the choice of a potent AMP, Lasioglossin-III (Lasio-III), to allow site specific immobilization of Lasio-III at an effective surface concentration. The Lasio-III peptide was chemically modified at the N-terminal with a cysteine residue to facilitate cysteine-directed immobilization of the peptide onto a commercial silicone catheter surface via a combination of an allyl glycidyl ether (AGE) brush and polyethylene glycol (PEG) based chemical coupling. The amount of immobilized peptide was determined to be 6.59 ± 0.89 µg cm-2 by Sulfo-SDTB assay. The AMP-coated catheter showed good antimicrobial activity against both Gram positive and negative bacteria. The antimicrobial properties of the AMP-coated catheter were sustained for at least 4 days post-incubation in a physiologically relevant environment and artificial urine and prevented the biofilm growth of E. coli and E. faecalis. Adenosine tri-phosphate leakage and propidium iodide fluorescence studies further confirmed the membranolytic mode of action of the immobilized peptide. To the best of our knowledge, this is the first proof-of-concept study that reports the efficacy of AMP immobilization by sulfhydryl coupling on a real catheter surface.

Immobilization studies of an engineered arginine-tryptophan-rich peptide on a silicone surface with antimicrobial and antibiofilm activity.

With the rapid rise of antibiotic-resistant-device-associated infections, there has been increasing demand for an antimicrobial biomedical surface. Synthetic antimicrobial peptides that have excellent bactericidal potency and negligible cytotoxicity are promising targets for immobilization on these target surfaces. An engineered arginine-tryptophan-rich peptide (CWR11) was developed, which displayed potent antimicrobial activity against a broad spectrum of microbes via membrane disruption, and possessed excellent salt resistance properties. A tethering platform was subsequently developed to tether CWR11 onto a model polymethylsiloxane (PDMS) surface using a simple and robust strategy. Surface characterization assays such as attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), and energy-dispersive X-ray spectroscopy (EDX) confirmed the successful grafting of CWR11 onto the chemically treated PDMS surface. The immobilized peptide concentration was 0.8 ± 0.2 µg/cm(2) as quantitated by sulfosuccinimidyl-4-o-(4,4-dimethoxytrityl) butyrate (sulfo-SDTB) assay. Antimicrobial assay and cytotoxic investigation confirmed that the peptide-immobilized surface has good bactericidal and antibiofilm properties, and is also noncytotoxic to mammalian cells. Tryptophan-arginine-rich antimicrobial peptides have the potential for antimicrobial protection of biomedical surfaces and may have important clinical applications in patients.

Immobilization of polybia-MPI by allyl glycidyl ether based brush chemistry to generate a novel antimicrobial surface.

Significant efforts towards the covalent immobilization of antimicrobial peptides (AMPs) on biomaterial surfaces are under way to render implantable biodevices with antimicrobial functionalities. Towards this aim, we studied the potential and effectiveness of a novel AMP candidate, polybia-MPI (Pmpi), for immobilization on a silicon substrate using allyl glycidyl ether (AGE) based polymerization chemistry. A statistical Design of Experiment (DoE) platform was developed to quantitatively understand the effects of different immobilization parameters on the final tethered peptide surface concentration. Our platform demonstrates efficient tethering of Pmpi up to ∼10 µg cm-2 on silicon wafers. Concentration dependent antimicrobial activities of the 'AGE-Pmpi'-tethered surfaces were observed, where a 70% reduction in bacterial colonies was achieved at a Pmpi surface concentration of 4.47 µg cm-2. The 'AGE-Pmpi'-tethered surfaces retained their antibacterial property after 3 days of incubation in artificial urine, and a 4-fold reduction in biofilm formation was also evident, as determined by ellipsometry. FESEM and ATP leakage assay showed that the immobilized Pmpi compromised bacterial membrane integrity but showed negligible cytotoxicity against human red blood cells.

High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies.

The Hepatitis B Virus X (HBx) protein is a potential therapeutic target for the treatment of hepatocellular carcinoma. However, consistent expression of the protein as insoluble inclusion bodies in bacteria host systems has largely hindered HBx manufacturing via economical biosynthesis routes, thereby impeding the development of anti-HBx therapeutic strategies. To eliminate this roadblock, this work reports the development of the first 'chromatography refolding'-based bioprocess for HBx using immobilised metal affinity chromatography (IMAC). This process enabled production of HBx at quantities and purity that facilitate their direct use in structural and molecular characterization studies. In line with the principles of quality by design (QbD), we used a statistical design of experiments (DoE) methodology to design the optimum process which delivered bioactive HBx at a productivity of 0.21 mg/ml/h at a refolding yield of 54% (at 10 mg/ml refolding concentration), which was 4.4-fold higher than that achieved in dilution refolding. The systematic DoE methodology adopted for this study enabled us to obtain important insights into the effect of different bioprocess parameters like the effect of buffer exchange gradients on HBx productivity and quality. Such a bioprocess design approach can play a pivotal role in developing intensified processes for other novel proteins, and hence helping to resolve validation and speed-to-market challenges faced by the biopharmaceutical industry today.

Transportal versus transtibial drilling technique of creating femoral tunnel in arthroscopic anterior cruciate ligament reconstruction using hamstring tendon autograft.

Drilling of femoral tunnel by transtibial technique is widely used in arthroscopic anterior cruciate ligament (ACL) reconstruction. Recent studies suggest in this technique graft is placed in non-anatomical position leading to instability. If the femoral tunnel is drilled through an anteromedial portal (transportal technique), graft can be placed more anatomically leading to better knee stability theoratically. The purpose of this study is to compare the clinical outcome of transtibial technique and transportal technique for drilling of femoral tunnel in arthroscopic ACL reconstruction using hamstring tendon autograft. All patients operated between January 2009 and September 2011 were approached for eligibility. Blinded assessment of IKDC score, Lachman test, pivot shift test, time of recovery from surgery were obtained from both the transtibial and transportal groups. The transportal group shows significantly better IKDC score, higher anteroposterior knee stability by Lachman test and lower recovery time from surgery.

Development of an enzyme-linked immunosorbent assay analytical platform for refolding yield determination of recombinant hepatitis B virus X (HBx) protein.

We report the development of a novel ELISA platform to quantitate hepatitis B virus X (HBx) protein refolding yields, which is critical for rational design and scaleup of aHBx bioprocess. HBx refolding yields were measured by determining the amount of HBx bound to immobilized GST-p53 on a "reduced glutathione"-functionalized maleimide surface. Refolding yields were distinguished from soluble yields, which were determined by measuring total HBx protein bound to a maleimide surface under reducing conditions. This platform is amenable to scaleup, and will expedite HBx production for structural and clinical studies, leading to the development of HBx-based therapy for liver cancer.

A rational design for hepatitis B virus X protein refolding and bioprocess development guided by second virial coefficient studies.

The hepatitis B virus X (HBx) protein is well known for its role in hepatitis B virus infection that often leads to hepatocellular carcinoma. Despite the clinical importance of HBx, there is little progress in anti-HBx drug development strategies due to shortage of HBx from native sources. Consistent expression of HBx as insoluble inclusion bodies within various expression systems has largely hindered HBx manufacturing via economical biosynthesis routes. Confronted by this roadblock, this study aims to quantitatively understand HBx protein behaviour in solution that will guide the rational development of a refolding-based bioprocess for HBx production. Second virial coefficient (SVC) measurements were employed to study the effects of varying physicochemical parameters on HBx intermolecular protein interaction. The SVC results suggest that covalent HBx aggregates play a key role in protein destabilisation during refolding. The use of an SVC-optimised refolding environment yielded bioactive and soluble HBx proteins from the denatured-reduced inclusion body state. This study provides new knowledge on HBx solubility behaviour in vitro, which is important in structure-function elucidation behaviour of this hydrophobic protein. Importantly, a rational refolding-based Escherichia coli bioprocess that can deliver purified and soluble HBx at large scale is successfully developed, which opens the way for rapid preparation of soluble HBx for further clinical and characterisation studies.

Exploring QSAR on 3-aminopyrazoles as antitumor agents for their inhibitory activity of CDK2/cyclin A.

Chemical inhibitors of cyclin-dependent kinases have great therapeutic potential against various proliferative and neurodegenerative disorders. The pharmacophoric requirement of 3-aminopyrazole, inhibitors of CDK2/cyclin A as antitumor agents was explored. QSAR study was performed using ETSA index, RTSA index, indicator parameters and atomic charges to consider quantitatively the effect of the structural variation on the antitumor activity of 3-aminopyrazole. Result showed that atom number 5 is important for the activity. It plays some electronic roles in the interaction of these compounds with enzymes as well as assumed to be involved through the dispersive/van der Waals interactions with enzyme. Presence of meta substitutions on the phenyl ring indicate the detrimental effects towards the activity. The presence of substituted biphenyl/2-thenyl phenyl at R1 are favorable towards the activity. QSAR study also indicates that with increasing the electronegativity of oxygen at position 8, the activity increases.