Automated analysis of single cells using Laser Tweezers Raman Spectroscopy.

In recent years, significant progress has been made into the label-free detection and discrimination of individual cancer cells using Laser Tweezers Raman Spectroscopy (LTRS). However, the majority of examples reported have involved manual trapping of cells, which is time consuming and may lead to different cell lines being analysed in discrete batches. A simple, low-cost microfluidic flow chamber is introduced which allows single cells to be optically trapped and analysed in an automated fashion, greatly reducing the level of operator input required. Two implementations of the flow chamber are discussed here; a basic single-channel device in which the fluid velocity is controlled manually, and a dual-channel device which permits the automated capture and analysis of multiple cell lines with no operator input. Results are presented for the discrimination of live epithelial prostate cells and lymphocytes, together with a consideration of the consequences of traditional 'batch analysis' typically used for LTRS of live cells.

Future directions for the European influenza reference laboratory network in influenza surveillance.

By defining strategic objectives for the network of influenza laboratories that have national influenza centre status or national function within European Union Member States, Iceland and Norway, it is possible to align their priorities in undertaking virological surveillance of influenza. This will help maintain and develop the network to meet and adapt to new challenges over the next 3-5 years and underpin a longer-term strategy over 5-10 years. We analysed the key activities undertaken by influenza reference laboratories in Europe and categorised them into a framework of four key strategic objectives areas: enhancing laboratory capability, ensuring laboratory capacity, providing emergency response and translating laboratory data into information for public health action. We make recommendations on the priority areas for future development.

Consensus recommendations for the use of FEIBA(®) in haemophilia A patients with inhibitors undergoing elective orthopaedic and non-orthopaedic surgery.

A growing number of publications have described the efficacy and safety of FEIBA as a first-line haemostatic agent for surgical procedures in haemophilia A patients with high-responding FVIII inhibitors. The aim of this study was to provide practical guidance on patient management and selection and also to communicate a standardized approach to the dosing and monitoring of FEIBA during and after surgery. A consensus group was convened with the aims of (i) providing an overview of the efficacy and safety of FEIBA in surgery; (ii) sharing best practice; (iii) developing recommendations based on the outcome of (i) and (ii). To date there have been 17 publications reporting on the use of FEIBA in over 210 major and minor orthopaedic and non-orthopaedic surgical procedures. Haemostatic outcome was rated as 'excellent' or 'good' in 78-100% of major cases. The reporting of thromboembolic complications or anamnestic response to FEIBA was very rare. Key to the success of FEIBA as haemostatic cover in surgery is to utilize the preplanning phase to prepare the patient both for surgery and also for rehabilitation. Haemostatic control with FEIBA should be continued for an adequate period postoperatively to support wound healing and to cover what can in some patients be an extended period of physiotherapy. Published data have demonstrated that FEIBA can provide adequate, well tolerated, peri and postoperative haemostatic cover for a variety of major and minor surgical procedures in patients with haemophilia A. The consensus recommendations provide a standardized approach to the dosing and monitoring of FEIBA.

Absorption spectroscopy in microfluidic flow cells using a metal clad leaky waveguide device with a porous gel waveguide layer.

Broadband absorption spectroscopy is advantageous because the full spectral profile of an analyte can permit identification of species. This work for the first time investigates the feasibility of a metal clad leaky waveguide (MCLW) device to obtain an absorption spectrum of an analyte of interest, methylene blue, using a white light source in a microfluidic flow cell. The MCLW device comprises a porous low refractive index gel, agarose, deposited on a titanium coated glass slide. The device was capable of detecting 2.3 µM of methylene blue at a wavelength of 650 nm. The corresponding minimum detectable absorbance is 1.6 × 10(-1) cm(-1). In comparison to commonly used detection devices the MCLW is simpler, robust, easier to fabricate and can be easily interfaced to microfluidic devices. It was also possible to store the MCLW devices dry for up to a year and rehydrate them in 30 s to a working condition.

Assessment of validity, reliability, responsiveness and bias of three commonly used patient-reported outcome measures in carpal tunnel syndrome.

BACKGROUND: In recent years there has been an increase in the use of self-administered questionnaires to accurately assess intervention outcomes in hand surgery in order to determine the quality of healthcare. This prospective study aims to evaluate and assess the validity, reliability, responsiveness, and bias of a number of outcome measure for Carpal Tunnel Syndrome (CTS) including the disease-specific Boston questionnaires (BQ), and the region-specific Disability of Arm, Shoulder, and Hand (DASH) questionnaires and Manchester Modified Disabilities of the Arm, Shoulder and Hand (M(2)DASH) questionnaires, and comparing the results to Nerve Conduction Studies (NCS). MATERIALS AND METHODS: Forty-eight patients with clinical signs of CTS confirmed by NCS completed the BQ, DASH and M(2)DASH questionnaire at different time intervals peri-operatively. The scores were analysed to assess validity, reliability, responsiveness, and bias of the questionnaires. Validity analysis for the three questionnaires showed strong positive correlations and there was no age, gender, hand dominance, or side affected bias in the questionnaires. RESULTS: No significant correlation was obtained between the questionnaires and NCS. Significant results for responsiveness were noted in BQ symptom severity scale only. CONCLUSION: 1. This study ha show n that the BQ, DASH and M(2)DASH questionnaires are valid and reliable outcome measures for CTS. 2. In terms of responsiveness, the DASH and M(2)DASH questionnaires are not as responsive as the BQ scores over the initial post-operative recovery period. 3. We would therefore recommend that the Boston Questionnaire be used to assess early post-operative patient related outcome measures for Carpal Tunnel Syndrome.

Microbiological aspects of public health planning and preparedness for the 2012 Olympic Games.

Although communicable diseases have hitherto played a small part in illness associated with Olympic Games, an outbreak of infection in a national team, Games venue or visiting spectators has the potential to disrupt a global sporting event and distract from the international celebration of athletic excellence. Preparation for hosting the Olympic Games includes implementation of early warning systems for detecting emerging infection problems. Ensuring capability for rapid microbiological diagnoses to inform situational risk assessments underpins the ability to dispel rumours. These are a prelude to control measures to minimize impact of any outbreak of infectious disease at a time of intense public scrutiny. Complex multidisciplinary teamwork combined with laboratory technical innovation and efficient information flows underlie the Health Protection Agency's preparation for the London 2012 Olympic and Paralympic Games. These will deliver durable legacies for clinical and public health microbiology, outbreak investigation and control in the coming years.

Musculoskeletal problems in persons with inhibitors: how do we treat?

Inhibitors are a serious complication, considerably increasing the morbidity, mortality and cost of treatment in this patient group. The challenge of treating people with haemophilia (PWH) with inhibitors can be met by a well-coordinated multidisciplinary team specialized in haemophilia. Each treatment centre must run a screening programme to detect inhibitors within their population and develop protocols to treat these patients. The treatment centre in Buenos Aires developed a screening programme that tests all our patients twice a year, ensuring early detection of inhibitors and early treatment of complications. In 2006, we analysed the quality of life (QOL) of non-inhibitor patients and compared it with inhibitor patients detected by this programme and found no differences in QOL measured by the SF36 questionnaire and no differences in school absenteeism. When diagnosis of the inhibitor does not come from a screening programme, its presence is suspected upon a lack of response to conventional replacement therapy for musculoskeletal bleeding, losing the 'golden moment' of treatment. This complication is much more serious when facing a traumatic bleed. In this situation, the lack of early diagnosis can lead to permanent damage or even death. Due to the cost of bypassing factors and the lack of experience of the medical team in the treatment of patients with inhibitors, many treatments that would improve the QOL of patients are instituted in an insufficient manner. Therefore, patients with haemophilia and inhibitors are often untreated or undertreated in their community. Orthopaedic surgeons and physiotherapists play a key role in the treatment of these patients and should be included in therapeutic decision making and most specifically in the postoperative treatment of patients with haemophilia and inhibitors. It is important that these patients have quick access to a trained therapeutic team in order to obtain an early diagnosis and treatment plan to prevent the evolution of the pathological process. Early treatment is cost-effective in maintaining and improving the QOL of patients. Experience in patients with haemophilia and inhibitors is not very extensive. Today, this situation is changing, with several treatment centres beginning to perform surgeries in these most complex patients, giving them a chance to improve their QOL. This article presents the experience of experts from various fields involved in treating patients with inhibitors from a developed and developing world perspective.

Management of the elbow joint.

The elbow is a complex joint that is prone to bleeding episodes. These features as well as the close proximity of the ulnar nerve and the need to use the elbow in many activities of daily living can lead to a range of symptoms including recurrent bleeds, pain, instability or loss of range of movement and nerve compression. Conservative management includes splinting and proprioceptive retraining monitored by a physiotherapist who is a musculoskeletal expert in hemophilia care. In the event that conservative measures are not successful a range of surgical options may be indicated including elbow replacement. These approaches continue to be evaluated in both the short and long term in order to determine the most effective treatment for the symptomatic elbow.

Carpal tunnel syndrome: a review of the recent literature.

Carpal Tunnel Syndrome (CTS) remains a puzzling and disabling condition present in 3.8% of the general population. CTS is the most well-known and frequent form of median nerve entrapment, and accounts for 90% of all entrapment neuropathies. This review aims to provide an overview of this common condition, with an emphasis on the pathophysiology involved in CTS. The clinical presentation and risk factors associated with CTS are discussed in this paper. Also, the various methods of diagnosis are explored; including nerve conduction studies, ultrasound, and magnetic resonance imaging.

Factor IX replacement to cover total knee replacement surgery in haemophilia B: a single-centre experience, 2000-2010.

Total knee replacement (TKR) is a well recognized treatment for haemophilic arthropathy. Successful haemostasis can be achieved by bolus doses or continuous infusion (CI) using either recombinant (r) or plasma-derived (pd) factor IX (FIX). We retrospectively analysed our experience of factor replacement to cover TKR in haemophilia B patients and explored factors related to FIX use during surgery. Between 2000 and 2010, 13 primary TKRs were performed in 11 haemophilia B patients. Operations were performed by the same surgeon using standard techniques. Median age was 58 years (42-79). An adjusted CI protocol was used for 5 days followed by bolus doses. FIX:C was maintained at 100 IU dL(-1) in the immediate postoperative period. There was no excess haemorrhage. There was no evidence of thrombosis or infection. All patients received mechanical thromboprophylaxis and only one chemical. CI was used in seven cases. Ten patients received pdFIX. Median hospital stay was 14 days (8-17). Median factor usage was 999 IU kg(-1) (768-1248). During CI, factor consumption was 695 IU kg(-1), 691 IU kg(-1) and 495 IU kg(-1) for BeneFix®, Replenine® and Haemonine, respectively. Clearance of both pdFIX and rFIX reduced during CI. All operations were uncomplicated. The decreased clearance in the CI setting reduced the amount of FIX required to maintain a therapeutic level. This reduction was greater with pdFIX and may be related to pharmacokinetic differences between pdFIX and rFIX. Given the excellent safety profile of the pdFIX products, CI of FIX and particularly pdFIX is safe, efficacious and convenient.

Perioperative clotting factor replacement and infection in total knee arthroplasty.

Total knee arthroplasty, or replacement (TKR), is now the most commonly performed surgical procedure performed in adults with haemophilia. It is indicated when end-stage haemophilic arthropathy results in intractable pain and reduced function. In patients with haemophilia, however, there has always been a concern about the high risk of infection, which carries with it potentially catastrophic consequences. The aims of this study were to review the case series of TKR for haemophilic arthropathy published in the medical literature, comparing the published infection rates and the differing clotting factor replacement regimes employed. Nineteen retrospective case series were identified; representing 556 TKR's in 455 patients with an overall infection rate of 7.9%. Case series which maintained a high level of clotting factor replacement throughout the first two postoperative weeks, however, had an infection rate of 2.15%, significantly lower than that of case series using the clotting factor replacement regime currently recommended in the World Federation of Hemophilia guidelines (9.22% P = 0.00545). We believe this study supports the use of a high level clotting factor replacement regime, replacing clotting factors to maintain them at a higher level for a longer period of time than currently recommended in international guidelines.

Genomic gain and over expression of CCL2 correlate with vascular invasion in stage I non-seminomatous testicular germ-cell tumours.

Testicular germ-cell tumours (TGCT) are the most frequent solid tumour to affect young Caucasian adult males and have increased in incidence over recent decades. In clinical stage I non-seminomas, (NSGCT) histological vascular invasion (VI) is a prognostic factor for metastatic relapse. Using array comparative genomic hybridization, we have previously shown that the presence of VI is associated with gain of a region at 17q12, containing a cluster of genes encoding inflammatory cytokines. We here confirm this finding using fluorescence in situ hybridization (FISH) demonstrating gain in 12 out of 42 (29%) assessable samples. Interrogation of previously published expression microarray data suggests that of the genes contained within this region, CCL2 [monocyte chemoattractant protein 1 (MCP1)] is frequently overexpressed in TGCT. Immunohistochemistry confirms this finding in a collection of 67 clinical stage I NSGCT, demonstrating an association with the presence of VI (p=0.049) that was not seen with VEGF-A, MMP2 or MMP9, although all were frequently expressed. This work gives further insight into the mechanisms involved in invasion in this tumour type, which may ultimately have implications for the management of patients with stage I disease.

Total knee replacement in patients with end-stage haemophilic arthropathy: 25-year results.

Haemophilia is an x-linked inherited bleeding disorder which can cause severe arthropathy. We have reviewed the results of 70 primary total knee replacements (TKR) performed in 57 haemophilic patients between 1983 and 2007. The functional results were assessed using the Hospital for Special Surgery (HSS) knee scoring system and Kaplan-Meier survivorship analysis. Six patients died. HSS scores were available for 60 TKRs at a mean follow-up of 9.2 years (2 to 23); 57 (95%) had good or excellent results. Deep infection was recorded in one patient. Kaplan-Meier analysis using infection and aseptic loosening as endpoints showed the survival rate at 20 years to be 94.0%. A reduction in infection, spontaneous haemarthrosis and improvement in the quality of life were noted to justify surgery in our series of patients with a mean age of 43 (25 to 70). We have found that using the latest techniques of continuous infusion of clotting Factor have significantly helped to reduce the complication rates and have achieved results which match those of the non-haemophilic population undergoing TKR.

Management of arthrofibrosis in haemophilic arthropathy.

SUMMARY: Hemophilic arthropathy is one of the conditions most associated with arthrofibrosis and loss of range of motion. Progressive fibrosis of synovium leads to pain, spasm, and shortening of muscles, resulting in joint contractures and restriction of joint motion. It is common to see even young children with severe loss of motion of elbows, knees and ankles. Treatment should be primarily by physiotherapy, splintage, and corrective devices. The late or severe cases may require surgical correction in the form of soft-tissue procedures, osteotomy and especially joint replacement.

Christiansen filter realized by an odd smooth cylindrical lens.

The Christiansen filter that is realized by odd smooth cylindrical lenses is analyzed in detail. Several popular filtering functions are discussed. The corresponding lens profile functions are obtained by an inverse scattering theory, which enables the filter to synthesize a desired prescribed response function. This kind of Christiansen filter has a passband narrower than that of the traditional Christiansen filter. Three Christiansen filters centered at 545 nm with full width at half-maximum of 2 nm are synthesized, and the approach to a better suppression of halos from the main transmission peak of the filters is presented in a systematic way.