.

Isopentenyl phosphate kinases (IPKs) catalyze the ATP-dependent phosphorylation of isopentenyl monophosphate (IP) to isopentenyl diphosphate (IPP) in the alternate mevalonate pathways of the archaea and plant cytoplasm. In recent years, IPKs have also been employed in artificial biosynthetic pathways called "(iso) prenol pathways" that utilize promiscuous kinases to sequentially phosphorylate (iso) prenol and generate the isoprenoid precursors IPP and dimethylallyl diphosphate (DMAPP). Furthermore, IPKs have garnered attention for their impressive substrate promiscuity toward non-natural alkyl-monophosphates (alkyl-Ps), which has prompted their utilization as biocatalysts for the generation of novel isoprenoids.

Transition from static culture to stirred tank bioreactor for the allogeneic production of therapeutic discogenic cell spheres.

Background: Culturing cells as cell spheres results in a tissue-like environment that drives unique cell phenotypes, making it useful for generating cell populations intended for therapeutic use. Unfortunately, common methods that utilize static suspension culture have limited scalability, making commercialization of such cell therapies challenging. Our team is developing an allogeneic cell therapy for the treatment of lumbar disc degeneration comprised of discogenic cells, which are progenitor cells expanded from human nucleus pulposus cells that are grown in a sphere configuration.

Chemoenzymatic synthesis of daptomycin analogs active against daptomycin-resistant strains.

Daptomycin is a last resort antibiotic for the treatment of infections caused by many Gram-positive bacterial strains, including vancomycin-resistant Enterococcus (VRE) and methicillin- and vancomycin-resistant Staphylococcus aureus (MRSA and VRSA). However, the emergence of daptomycin-resistant strains of S. aureus and Enterococcus in recent years has renewed interest in synthesizing daptomycin analogs to overcome resistance mechanisms.

Acceptor substrate determines donor specificity of an aromatic prenyltransferase: expanding the biocatalytic potential of NphB.

Aromatic prenyltransferases are known for their extensive promiscuity toward aromatic acceptor substrates and their ability to form various carbon-carbon and carbon-heteroatom bonds. Of particular interest among the prenyltransferases is NphB, whose ability to geranylate cannabinoid precursors has been utilized in several in vivo and in vitro systems. It has therefore been established that prenyltransferases can be utilized as biocatalysts for the generation of useful compounds.

FgaPT2, a biocatalytic tool for alkyl-diversification of indole natural products.

Aromatic prenyltransferases from natural product biosynthetic pathways display relaxed specificity for their aromatic substrates. While a growing body of evidence suggests aromatic prenyltransferases to be more tolerant towards their alkyl-donor substrates, most studies aimed at probing their donor-substrate specificity are limited to only a small set of alkyl pyrophosphate donors, restricting their broader utility as biocatalysts for synthetic applications. Here, we assess the donor substrate specificity of an l-tryptophan C4-prenyltransferase, also known as C4-dimethylallyltryptophan synthase, FgaPT2 from , using an array of 34 synthetic unnatural alkyl-pyrophosphate analogues, and demonstrate FgaPT2 can catalyze the transfer of 25 of the 34 non-native alkyl groups from their corresponding synthetic alkyl-pyrophosphate analogues at N1, C3, C4 and C5 position of tryptophan in a normal and reverse manner.

Determination of Alkyl-Donor Promiscuity of Tyrosine-O-Prenyltransferase SirD from Leptosphaeria maculans.

Natural product prenyltransferases are known to display relaxed acceptor substrate specificity. Although recent studies with a small set of unnatural alkyl donors have revealed that prenyltransferases are flexible with regard to their alkyl donors, the scope of their alkyl donor specificity remains poorly understood. Towards this goal, we report the synthesis of 20 unnatural alkyl pyrophosphate donors and an assessment of the reactions of these synthetic unnatural alkyl pyrophosphate analogues catalyzed by tyrosine O-prenyltransferase SirD.

Efficacy of ampicillin against methicillin-resistant Staphylococcus aureus restored through synergy with branched poly(ethylenimine).

β-Lactam antibiotics kill Staphylococcus aureus bacteria by inhibiting the function of cell wall penicillin-binding proteins (PBPs) 1 and 3. However, β-lactams are ineffective against PBP2a, used by methicillin-resistant S. aureus (MRSA) to perform essential cell wall crosslinking functions.

A case-control study investigating factors of preoperative delay in emergency laparotomy.

Background: Emergency laparotomy (EL) is a procedure that puts a strain on healthcare resources and is associated with a significant morbidity and mortality. Despite these implications little improvement in the outcome of patients undergoing this procedure has been made in the UK over the last few decades. A delay in transferring patients to theatre has been shown to negatively affect outcome of EL.

Bacterial lipoteichoic acid enhances cryosurvival.

Antifreeze proteins in fish, plants, and insects provide protection to a few degrees below freezing. Microbes have been found to survive at even lower temperatures, and with a few exceptions, antifreeze proteins are missing. We show that lipoteichoic acid (LTA), a biopolymer in the cell wall of Gram-positive bacteria, can be added to B.

Musculoskeletal effects of an electrical stimulation induced cycling programme in the spinal injured.

Twelve patients were involved in a 3 month stimulation induced cycling programme at the Royal Perth Rehabilitation Hospital. A number of the patients were less than 1 year post injury, all except one had an incomplete injury, and most were receiving physiotherapy. All patients who completed the programme increased their time of cycling and, in all but one case, the exercise load, indicative of a local training effect.

A clinical exercise system for paraplegics using functional electrical stimulation.

A low cost clinical exercise system was developed for the spinal cord injured, based on a bicycle ergometer and electrical stimulation. A pilot project was conducted, using the system, to examine the effects of stimulation induced cycling in long term paraplegics. The project comprised 2 phases of exercise, a strengthening phase involving a 12 week programme of electrical stimulation to the quadriceps and hamstrings and a 12 week cycling phase.

Reliability of the Cobb angle index derived by traditional and computer assisted methods.

The reliability of the traditional Cobb angle index, which at present exhibits a wide range of reported values, was determined in this study and amounted to a within subject error of +/-4.3 degrees and a between subject error of +/-4.9 degrees.

A device for producing experimental fractures.

A device is reported for the production of transverse fractures of canine tibiae by a three-point bending technique. With strain-gauged load arms, the device enables simultaneous measurement of the bend strength of the intact bone. Results from a series of 14 dogs confirm the reproducibility of this technique.

An improved orthopaedic stirrup.

A single RPH universal stirrup covers the many types of traction needed in orthopaedic practice. It reduces the torque load on the Steinmann pin, thereby diminishing the rate of loosening of the pin in the bone. From our experience with the stirrup at the Royal Perth Hospital, savings in ward costs can be achieved by shortening the time in hospital and reducing the need for antibiotic cover, by decreasing maintenance costs and by reducing the size of the appliance stock kept to cover the range of tractions used in orthopaedic practice.

The dynamic mechanical response characteristics of spinal cord tissue: a preliminary report.

In the absence of an adequate description of the response of spinal cord to dynamic insult, work is in hand to determine the dynamic characteristics of canine spinal cord tissue. In-vitro experimentation is described. A vibration technique using a low amplitude (0.