The discovery and optimization of benzimidazoles as selective NaV1.8 blockers for the treatment of pain.

The voltage gated sodium channel NaV1.8 has been postulated to play a key role in the transmission of pain signals. Core hopping from our previously reported phenylimidazole leads has allowed the identification of a novel series of benzimidazole NaV1.8 blockers. Subsequent optimization allowed the identification of compound 9, PF-06305591, as a potent, highly selective blocker with an excellent preclinical in vitro ADME and safety profile.

The Use of Routine Postoperative Microscopy and Culture Screening Following Elective Hip and Knee Arthroplasty: An Unnecessary Cost With No Effect on Clinical Management?

BACKGROUND: The use of microscopy and culture screening to detect pathogenic microorganisms followed by a decolonization protocol is a widely performed practice prior to elective hip and knee arthroplasty. In our center, the routine care of hip and knee arthroplasty also involves postoperative screening including direct culture of the surgical site. The aim of this study was to assess the frequency of pathogen detection following these tests and to determine whether routine postoperative screening, with particular reference to postoperative surgical site culture, led to any change in clinical management of these patients. METHODS: A series of 1000 patients undergoing hip or knee arthroplasty at The Mater Hospital between January 2014 and December 2015 were identified from our arthroplasty database. Results of preoperative and postoperative microscopy and culture screening were reviewed by 2 independent researchers. RESULTS: Of the 1000 subjects, positive microscopy and culture results were identified in 88 patients (8.8%) preoperatively and 5 patients (0.5%) postoperatively. None of the 1000 postoperative surgical site swabs had a positive microscopy and culture screen. All the 5 positive postoperative microscopy and culture screen results were in patients who had positive cultures preoperatively. There were no positive postoperative microscopy and culture screen results in patients who had had negative preoperative results. Postoperative screening was performed at a cost of AUS$213 per patient. CONCLUSION: Routine postoperative surgical site culture following hip and knee arthroplasty does not alter clinical management, has a significant associated financial cost, and has the potential to expose the patient to a risk of surgical site infection and is therefore not supported.

The long-term outcome of the Gschwend-Scheier-Bähler III elbow replacement.

BACKGROUND: The Gschwend-Scheier-Bähler III (GSBIII) is a semiconstrained, sloppy-hinge total elbow replacement. We report the long-term functional and radiological outcome of a cohort of patients more than 10 years after surgery. METHODS: All GSBIII prostheses implanted from September 1996 to June 2004 were identified from our surgical database. Functional and radiological assessments were performed at routine patient clinic visits, using the Oxford Elbow Score, the 11-item version of the Disabilities of Arm, Shoulder and Hand score (QuickDASH), and plain radiographs. RESULTS: From 1996 to 2004, 52 elbows in 40 patients were implanted; of these, 18 patients (23 elbows) had died, leaving 22 patients with 29 elbows available for follow-up. Three patients (3 elbows) could not be contacted. Functional and radiological data were available for 19 patients with 26 elbows (90%). Overall survival was a mean of 13.1 years (range, 10.6-16.4 years). Mean age at operation was 63.0 years (range, 49.5-80.6 years). There were 5 male elbows and 21 female elbows. Five total elbow replacements were performed for osteoarthritis and 24 for rheumatoid arthritis. The mean Oxford Elbow Score was 26.9 (range, 18-48). The mean QuickDASH score was 42.6 (range, 2.5-93.2). Of the 52 elbows in 40 patients, 4 elbows (7.7%) required further surgery, 2 (3.8%) of which were revisions. In addition, there was 1 intraoperative complication and 2 postoperative complications not requiring further surgery. Kaplan-Meier 10-year survival shows a 95.9% implant survival with revision as the end point. CONCLUSIONS: The GSBIII elbow replacement provides good long-term function with a low revision rate and few complications. LEVEL OF EVIDENCE: Level IV; Case Series; Treatment Study.

Complications after revision surgery of malreduced ankle fractures.

Ankle fractures are common orthopedic injuries requiring reduction and cast immobilization or fixation. Fractures fixed in a malreduced (misaligned) position can require revision surgery. However, because this has been a relatively rare occurrence, little is known about the complications that can occur after such surgery. We reviewed all adult closed ankle fractures that underwent revision surgery for technical failure in a regional trauma hospital from January 2007 to January 2010. Those with open fractures and those who required external fixation at any point in their treatment were excluded. Nine patients underwent revision surgery during the study period. Of these 9 patients, 3 (33%) developed a deep infection, all with positive microbiology cultures for methicillin-sensitive Staphylococcus aureus. Each of these patients underwent removal of the metalwork and wound debridement, followed by plastic surgery free flap coverage. In addition to the 3 infection cases, those with noninfected complications included 1 patient (11%) with chronic regional pain syndrome, 1 (11%) with failure of plate fixation, and 1 (11%) with persistent pain requiring arthroscopy and debridement. The overall incidence of complications was 66.67% in this group of 9 patients who had undergone revision surgery for the treatment of a malreduced malleolar ankle fracture. Although our observational study involved a small subset of patients who had undergone surgical repair for ankle fracture, we suggest that if revision surgery will be undertaken, the high incidence of infection and the potential need for plastic surgery should be highlighted during the consent process before the original, open reduction internal fixation procedure.

Knockout or inhibition of USP30 protects dopaminergic neurons in a Parkinson's disease mouse model.

Mutations in SNCA, the gene encoding α-synuclein (αSyn), cause familial Parkinson's disease (PD) and aberrant αSyn is a key pathological hallmark of idiopathic PD. This α-synucleinopathy leads to mitochondrial dysfunction, which may drive dopaminergic neurodegeneration. PARKIN and PINK1, mutated in autosomal recessive PD, regulate the preferential autophagic clearance of dysfunctional mitochondria ("mitophagy") by inducing ubiquitylation of mitochondrial proteins, a process counteracted by deubiquitylation via USP30. Here we show that loss of USP30 in Usp30 knockout mice protects against behavioral deficits and leads to increased mitophagy, decreased phospho-S129 αSyn, and attenuation of SN dopaminergic neuronal loss induced by αSyn. These observations were recapitulated with a potent, selective, brain-penetrant USP30 inhibitor, MTX115325, with good drug-like properties. These data strongly support further study of USP30 inhibition as a potential disease-modifying therapy for PD.

Functional and oncological outcome following marginal excision of well-differentiated forearm liposarcoma with nerve involvement.

PURPOSE: Liposarcoma is one of the most common soft tissue sarcomas in adults. It is often low-grade and can occasionally involve neurovascular structures. We present the functional and oncological outcome resulting from planned marginal excision of a series of forearm low-grade liposarcomas with nerve involvement. METHODS: The Oxford tumor registry was used to identify cases of histologically proven, well-differentiated liposarcoma of the forearm, with nerve involvement, treated surgically between 1997 and 2006. Nerve involvement was identified clinically with symptoms or signs of nerve compression, or by images showing direct contact of the tumor with a nerve on magnetic resonance imaging. This was then further defined at the time of surgery as tumor abutting (capsular involvement) or encasing a peripheral nerve. Demographic and clinical data were collected and oncological outcome was assessed by noting local and distant recurrence during follow-up. Postoperative functional outcome was assessed using the Toronto Extremity Salvage Scores. RESULTS: Eight cases were identified, 6 with preoperative neurological symptoms. The total group comprised 6 men and 2 women with a mean age of 61 (range, 30-71) years. At surgery, all had their tumors successfully excised, with preservation of the involved nerves. In those with preoperative neurological symptoms, complete recovery occurred by 18 months after surgery. The average follow-up was 5 years (range, 3-9 y). There were no cases of either local or distant recurrence of disease, with a mean Toronto Extremity Salvage Score of 99%. CONCLUSIONS: Planned marginal excision of a well-differentiated liposarcoma, arising in the forearm and involving nerve, can result in excellent functional and oncological outcome. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV.

Avulsion of the perforating branch of the peroneal artery secondary to an ankle sprain: a cause of acute compartment syndrome in the leg.

In this report, we describe the case of an adult male who developed an acute compartment syndrome localized to the anterior compartment of the leg following an ankle sprain. Compartment syndrome in association with ankle sprain is unusual, and has been previously described in association with avulsion of the perforating peroneal artery. Because of the potential for severe morbidity, we feel that it is important to make foot and ankle surgeons aware of this unusual injury.

Modulation of the partition coefficient between octanol and buffer at pH 7.4 and pKa to achieve the optimum balance of blood clearance and volume of distribution for a series of tetrahydropyran histamine type 3 receptor antagonists.

The relationship between rat pharmacokinetics and physicochemical parameters [the partition coefficient between octanol and buffer at pH 7.4 (log D((7.4))) and pK(a)] was studied for a series of tetrahydropyran compounds. Sixteen compounds ranging in log D((7.4)) 0.1 to 1.8 were administered intravenously to rats, and the pharmacokinetic parameters were determined from blood concentration time curves. Across the series, a weak correlation was observed between log D((7.4)) and blood clearance, suggesting that log D((7.4)) values less than 0.5 were required to prevent clearance at hepatic blood flow. In terms of the volume of distribution (V(d)), the compounds fell into three distinct subseries characterized by the number of basic centers and differences in ionization of each basic center at physiological pH. These were referred to as the monobasic, weak second base, and strong second base subseries. All the compounds exhibited V(d) greater than body water, as would be expected from their lipophilic and basic nature. For a given clog P, the strong second base subseries showed higher V(d) than the weak second base subseries, which in turn exhibited higher values than the monobasic subseries. In addition, for the weak second base subseries, V(d) could be tuned by modulating the pK(a) of the second basic center. This relationship was rationalized in respect to the interactions of the ionizable centers with phospholipid heads in the cell membrane and/or lysosomal trapping. Compounds in the weak second base subseries showed optimal V(d), and when combined with a log D((7.4)) of 0.1, driving to moderate blood clearance, one compound showed the optimal pharmacokinetic profile.

Does the femoral offset affect replacements? The results from a National Joint Registry.

BACKGROUND: Femoral component offset influences the torque forces exerted on a femoral stem and may therefore adversely affect femoral component survival. This study investigated the influence of femoral component offset on revision rates for primary total hip replacements (THR) registered on the New Zealand Joint Registry (NZJR). METHODS: There were 106,139 primary THRs registered, resulting in 4960 revisions for any cause. There were 46,242 THRs performed using the five commonest femoral components listed on the NZJR. A total of 41,100 were done for primary osteoarthritis of which 40,548 had all the offset information available for analysis. We defined low offset as < 42 mm, standard as 42-48 mm and high offset as > 48 mm offset and examined revision rates according to the reasons for revision. We performed survival analyses for both cemented and uncemented femoral components grouped by the different offsets. RESULTS: The all-cause revision rate was 0.54/100 component years (cys). Stems with < 42 mm offset had a revision rate of 0.58/100 cys (mean 0.58; 95% confidence interval (CI) 0.53-0.63), 42-48 mm offset 0.47 (95% CI 0.43-0.52) and > 48 mm offset 0.67 (95% CI 0.57-0.79). There was no significant difference in all-cause revision rates between varying stem offsets in uncemented stems adjusting for age and gender. In cemented stems both high and low offset stems were more likely to be revised. Uncemented stems of all offsets were more likely to undergo revision for femoral fracture. CONCLUSIONS: Femoral component offset affects the overall all-cause revision rate of the most commonly used cemented stem, but not uncemented stem designs. In cemented stems offset influences the rate of revision for loosening and periprosthetic fractures.

A high Q piezoelectric resonator as a portable VLF transmitter.

Very low frequency communication systems (3 kHz-30 kHz) enable applications not feasible at higher frequencies. However, the highest radiation efficiency antennas require size at the scale of the wavelength (here, >1 km), making portable transmitters extremely challenging. Facilitating transmitters at the 10 cm scale, we demonstrate an ultra-low loss lithium niobate piezoelectric electric dipole driven at acoustic resonance that radiates with greater than 300x higher efficiency compared to the previous state of the art at a comparable electrical size. A piezoelectric radiating element eliminates the need for large impedance matching networks as it self-resonates at the acoustic wavelength. Temporal modulation of this resonance demonstrates a device bandwidth greater than 83x beyond the conventional Bode-Fano limit, thus increasing the transmitter bitrate while still minimizing losses. These results will open new applications for portable, electrically small antennas.

Does overstuffing of the patellofemoral joint in total knee arthroplasty have a significant effect on postoperative outcomes?

BACKGROUND: There is ongoing debate in the literature as to whether or not patellofemoral joint overstuffing has a clinically significant effect on postoperative outcomes following total knee arthroplasty (TKA). This study investigates the effect of patellofemoral joint overstuffing on patient-reported outcomes using novel methods of radiographic measurement. METHODS: The study population consisted of a prospective cohort of 266 patients receiving a Triathlon® (Stryker, Kalamazoo, MI, USA) TKA between 2006 and 2009. Participants completed the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) questionnaire preoperatively and at 12 months postoperatively. Pre- and postoperative radiographic measurements were taken according to a defined protocol to assess for patellofemoral overstuffing. Measurement reproducibility was assessed using inter-observer intraclass correlation coefficients. Associations between radiographic measurements and patient-reported outcomes were analysed using linear regression analysis. RESULTS: A total of 107 patients had adequate images and were included in the analysis for this study. Three different radiographic measurements were used to identify patellofemoral overstuffing all with good intra- and inter-observer reliability. There was no association identified between combined (patella and trochlea) patellofemoral overstuffing measurements and WOMAC scores. However, a statistically significant association was identified between an increase in anterior trochlear offset and worse knee pain and function scores (P < 0.05). CONCLUSIONS: There is no identifiable association between true patellofemoral overstuffing and clinical outcome; however, there is a small association with the anterior trochlear offset though further studies are warranted to confirm the clinical significance of this finding.

Subtype-specific actions of beta-amyloid peptides on recombinant human neuronal nicotinic acetylcholine receptors (alpha7, alpha4beta2, alpha3beta4) expressed in Xenopus laevis oocytes.

Two-electrode voltage-clamp electrophysiology has been used to study the actions of two amyloid peptides (Abeta(1-42), Abeta(1-40)) on alpha7, alpha4beta2 and alpha3beta4 recombinant human neuronal nicotinic acetylcholine receptors (nicotinic AChRs), heterologously expressed in Xenopus laevis oocytes. The application of Abeta(1-42) or Abeta(1-40) (1 pM-100 nM) for 5 s does not directly activate expressed human alpha7, alpha4beta2 or alpha3beta4 nicotinic AChRs.Abeta(1-42) and Abeta(1-40) are antagonists of alpha7 nicotinic AChRs. For example, 10 nM Abeta(1-42) and Abeta(1-40) both reduced the peak amplitude of currents recorded (3 mM ACh) to 48+/-5 and 45+/-10% (respectively) of control currents recorded in the absence of peptide. In both the cases the effect is sustained throughout a 30 min peptide application and is poorly reversible.Abeta(1-42) and Abeta(1-40) (10 nM) enhance currents recorded in response to ACh (3 mM) from oocytes expressing alpha4beta2 nicotinic AChRs by 195+/-40 and 195+/-41% respectively. This effect is transient, reaching a peak after 3 min and returning to control values after a 24 min application of 10 nM Abeta(1-42). We observe an enhancement of 157+/-22% of control ACh-evoked current amplitude in response to 100 nM Abeta(1-42) recorded from oocytes expressing alpha4beta2 nicotinic AChRs.Abeta(1-42) and Abeta(1-40) (10 nM) were without antagonist actions on the responses of alpha3beta4 nicotinic AChRs to ACh (1 nM-3 mM).

Discovery and Optimization of Selective Nav1.8 Modulator Series That Demonstrate Efficacy in Preclinical Models of Pain.

Voltage-gated sodium channels, in particular Nav1.8, can be targeted for the treatment of neuropathic and inflammatory pain. Herein, we described the optimization of Nav1.8 modulator series to deliver subtype selective, state, and use-dependent chemical matter that is efficacious in preclinical models of neuropathic and inflammatory pain.

A novel selective and orally bioavailable Nav 1.8 channel blocker, PF-01247324, attenuates nociception and sensory neuron excitability.

BACKGROUND AND PURPOSE: NaV 1.8 ion channels have been highlighted as important molecular targets for the design of low MW blockers for the treatment of chronic pain. Here, we describe the effects of PF-01247324, a new generation, selective, orally bioavailable Nav 1.8 channel blocker of novel chemotype. EXPERIMENTAL APPROACH: The inhibition of Nav 1.8 channels by PF-01247324 was studied using in vitro patch-clamp electrophysiology and the oral bioavailability and antinociceptive effects demonstrated using in vivo rodent models of inflammatory and neuropathic pain. KEY RESULTS: PF-01247324 inhibited native tetrodotoxin-resistant (TTX-R) currents in human dorsal root ganglion (DRG) neurons (IC50 : 331 nM) and in recombinantly expressed h Nav 1.8 channels (IC50 : 196 nM), with 50-fold selectivity over recombinantly expressed TTX-R hNav 1.5 channels (IC50 : ∼10 µM) and 65-100-fold selectivity over TTX-sensitive (TTX-S) channels (IC50 : ∼10-18 µM). Native TTX-R currents in small-diameter rodent DRG neurons were inhibited with an IC50 448 nM, and the block of both human recombinant Nav 1.8 channels and TTX-R from rat DRG neurons was both frequency and state dependent. In vitro current clamp showed that PF-01247324 reduced excitability in both rat and human DRG neurons and also altered the waveform of the action potential. In vivo experiments n rodents demonstrated efficacy in both inflammatory and neuropathic pain models. CONCLUSIONS AND IMPLICATIONS: Using PF-01247324, we have confirmed a role for Nav 1.8 channels in both inflammatory and neuropathic pain. We have also demonstrated a key role for Nav 1.8 channels in action potential upstroke and repetitive firing of rat and human DRG neurons.

Deubiquitylating enzymes and DNA damage response pathways.

Covalent post-translational modification of proteins by ubiquitin and ubiquitin-like factors has emerged as a general mechanism to regulate myriad intra-cellular processes. The addition and removal of ubiquitin or ubiquitin-like proteins from factors has recently been demonstrated as a key mechanism to modulate DNA damage response (DDR) pathways. It is thus, timely to evaluate the potential for ubiquitin pathway enzymes as DDR drug targets for therapeutic intervention. The synthetic lethal approach provides exciting opportunities for the development of targeted therapies to treat cancer: most tumours have lost critical DDR pathways, and thus rely more heavily on the remaining pathways, while normal tissues are still equipped with all DDR pathways. Here, we review key deubiquitylating enzymes (DUBs) involved in DDR pathways, and describe how targeting DUBs may lead to selective therapies to treat cancer patients.