Sphyga: a multiparameter open source tool for fabricating smart and tunable hydrogel microbeads.

Hydrogel microbeads are used in many biological applications, particularly for cell, protein or drug encapsulation. Although there are several methods for fabricating microbeads with controlled shapes and dimensions, many are limited to a small range of materials or sizes. We describe a compact open source tool-the spherical hydrogel generator (Sphyga)-for the fabrication of highly reproducible hydrogel based microbeads with predictable shapes and diameters ranging from 100 to 2000 µm. The unique feature of the system is the ability to modulate multiple parameters independently, so as to create a wide range of working conditions for fabricating tailored microbeads. Hence, by combining the different fabrication parameters, hydrogel beads with chosen shapes, sizes and materials can be generated with Sphyga. A multiparameter working-window was obtained by fixing the concentration of the base material, alginate, and varying the viscosity of the solution along with Sphyga's fabrication parameters (needle size, external air pressure, and material outflow). To validate the multiparameter working window, components such as proteins, cells, dyes and nanoparticles were also used to fabricate composite microbeads. The results show that the architecture of hydrogel microbeads can be engineered by considering the viscosity of the initial solution, which depends principally on the pH and composition of alginate solution. Coupled with Sphyga's multiple working parameters, material viscosity can then be used to tune hydrogel domains and thereby generate complex biologically relevant microenvironments for many biomedical applications.

A new device to measure the three dimensional forces and torques in precision grip.

Fine prehension is ubiquitous in everyday skilled hand manipulations. The anticipatory nature of the control of normal grip forces exerted against tangential loads has been extensively used to provide insights into the working of neural control of movements. We designed a new versatile device to measure the three dimensional forces and torques during a broad panel of precision grip tasks. The instrument measures constraints exerted independently on two grip surfaces by the thumb and the opposing fingers. In addition, the device can be loaded to increase the weight and/or induce torques and can be easily integrated in a variety of experimental contexts. Its compactness improves its stability during movement and allows an ergonomic manipulation for impaired persons or children.

Functional assessment in physiotherapy. A literature review.

The present literature review on functional assessment in physiotherapy was carried out for the following reasons: 1) to identify the functional instruments used in the field of physiotherapy that were supported by published evidence of their psychometric qualities; 2) to investigate how these instruments relate to the International Classification of Functioning, Disability and Health (ICF); and 3) to investigate the use of functional instruments in the financing of physiotherapy. A search of Medline from 1990 to December 2005, in the domains of functional evaluation, psychometric qualities, functional classification, and health policy in relation to physiotherapy resulted in a list of 1,567 studies. Two reviewers examined the resulting references on the basis of their title and abstract, in order to select the studies that presented data on the psychometric qualities of functional evaluation tests, leading to a final selection of 44 such studies. A selection of functional tests was identified in four major diagnostic groups treated in community physiotherapy: musculoskeletal disorders (including lower back pain), stroke, the elderly, and traumatic brain injuries. The functional tests authors identified essentially cover the body and activities dimension of the ICF. The selected tests could be used as a basis for the standardisation of functional evaluation of the major diagnostic groups treated in community physiotherapy. This means that standards are available for reporting and following the evolution of patients both longitudinally and transversally. Nevertheless, in the current literature review no attempt at using functional outcomes as a rationale for financing physiotherapy could be found to date.

Listeria monocytogenes in a young patient with non Hodgkins lymphoma: case report.

Listeria monocytogenes is an intracellular food-borne pathogen, widely distributed in the environment, which rarely causes clinical infection in healthy people, but may cause severe disease in immunocompromised patients. A case of listeriosis is certified in an immunocompromised patient, thus confirming this microorganism to be an opportunistic human pathogen.

Gut-associated bacterial microbiota in paediatric patients with inflammatory bowel disease.

BACKGROUND: Clinical and experimental observations in animal models indicate that intestinal commensal bacteria are involved in the initiation and amplification of inflammatory bowel disease (IBD). No paediatric reports are available on intestinal endogenous microflora in IBD. AIMS: To investigate and characterise the predominant composition of the mucosa-associated intestinal microflora in colonoscopic biopsy specimens of paediatric patients with newly diagnosed IBD. METHODS: Mucosa-associated bacteria were quantified and isolated from biopsy specimens of the ileum, caecum and rectum obtained at colonoscopy in 12 patients with Crohn's disease, 7 with ulcerative colitis, 6 with indeterminate colitis, 10 with lymphonodular hyperplasia of the distal ileum and in 7 controls. Isolation and characterisation were carried out by conventional culture techniques for aerobic and facultative-anaerobic microorganisms, and molecular analysis (16S rRNA-based amplification and real-time polymerase chain reaction assays) for the detection of anaerobic bacterial groups or species. RESULTS: A higher number of mucosa-associated aerobic and facultative-anaerobic bacteria were found in biopsy specimens of children with IBD than in controls. An overall decrease in some bacterial species or groups belonging to the normal anaerobic intestinal flora was suggested by molecular approaches; in particular, occurrence of Bacteroides vulgatus was low in Crohn's disease, ulcerative colitis and indeterminate colitis specimens. CONCLUSION: This is the first paediatric report investigating the intestinal mucosa-associated microflora in patients of the IBD spectrum. These results, although limited by the sample size, allow a better understanding of changes in mucosa-associated bacterial flora in these patients, showing either a predominance of some potentially harmful bacterial groups or a decrease in beneficial bacterial species. These data underline the central role of mucosa-adherent bacteria in IBD.

Encrusted cystitis in an immunocompromised patient: possible coinfection by Corynebacterium urealyticum and E. coli.

Encrusted cystitis is a severe chronic inflammatory disease of the bladder characterized by excessively alkaline urine and calcifications within the bladder wall. A case of a 60 year-old man affected by systemic lupus erythematosus (SLE), which developed encrusted cystitis due to Corynebacterium urealyticum with E. coli co-infection, shows that the treatment of encrusted cystitis with a endoscopic debulking of the encrusted stones and an antimicrobial therapy specific for C. urealyticum often is not sufficient for the complete resolution of symptoms.

Cross-cultural validity of FIM in spinal cord injury.

OBJECTIVE: To analyse cross-culture validity of the Functional Independence Measure (FIM) in patients with a spinal cord injury using a modern psychometric approach. SETTINGS: A total of 19 rehabilitation facilities from four countries in Europe. PARTICIPANTS: A total of 647 patients at admission, median age 46 years, 69% male. METHODS: Data from the FIM, collected on inpatient admission, was fitted to the Rasch model. A detailed analysis of scoring functions of the seven categories of the FIM items was undertaken before to testing fit to the model. Categories were rescored where necessary. Fit to the model was assessed initially within country, and then in the pooled data. Analysis of differential item functioning (DIF) was undertaken in the pooled data for each of the FIM motor and social cognitive scales, respectively. Final fit to the model was tested for breach of local independence by principle components analysis (PCA). RESULTS: The present scoring system for the FIM motor and cognitive scales, that is a seven category scale, was found to be invalid, necessitating extensive rescoring. Following this, DIF was found in a number of items within the motor scale, requiring a complex solution of splitting items by country to allow for the valid pooling of data. Five country-specific items could not be retained within this solution. The FIM cognitive scale fitted the Rasch model after rescoring, but there was a substantial ceiling effect. CONCLUSIONS: Data from the FIM motor scale for patients with spinal cord injury should not be pooled in its raw form, or compared from country to country. Only after fit to the Rasch model and necessary adjustments could such a comparison be made, but with a loss of clinical important items. The FIM cognitive scale works well following rescoring, and data may be pooled, but many patients were at the maximum score.

Apoptotic death of Listeria monocytogenes-infected human macrophages induced by lactoferricin B, a bovine lactoferrin-derived peptide.

Listeria monocytogenes, an intracellular facultative food-borne pathogen, was reported to induce apoptosis in vitro and in vivo in a variety of cell types with the exception of murine macrophages. These cells represent the predominant compartment of bacterial multiplication and die as a result of necrosis. In this study we showed that human non-activated and IFN-gamma-activated macrophagic-like (THP-1) cells infected with L. monocytogenes, mainly die by necrosis rather than by an apoptotic process. Two natural products derived from bovine milk, lactoferrin and its derivative peptide lactoferricin B, are capable of regulating the fate of infected human macrophages. Bovine lactoferrin treatment of macrophages protects them from L. monocytogenes-induced death whereas lactoferricin B, its derivative peptide, determines a shifting of the equilibrium from necrosis to apoptosis.

Interstitial cystitis and infectious agents.

Interstitial cystitis (IC) is a syndrome consisting of severe refractory bladder symptoms of unknown etiology. The disease tends to affect Caucasian women with a mean age of 40 years, with 25% of patients under the age of 30. Few population based epidemiological studies of IC have been performed. We analyzed a case of interstitial cystitis in a 42-year-old non-smoker woman. In two biopsy samples the presence of viral DNA of human polyomavirus BK (BKV), human herpes virus type 1 and type 2 (HHV- 1 and HHV-2), adenovirus, human papillomavirus (HPV) and bacterial DNA (Chlamydia trachomatis and Mycoplasma genitalium) were evaluated by means of polymerase chain reaction (PCR). Both samples resulted positive only for BKV and HPV DNA. HPV genotyping revealed the presence of HPV-66 that is associated with a high risk of cancer development. Thus the finding of a viral co-infection could support the hypothesis of the multi-factorial origin of this pathology.

Infectious agents in tissues from spontaneous abortions in the first trimester of pregnancy.

Some evidence suggests that intrauterine infection plays a major role in the pathogenesis of early pregnancy loss, but the implication and prevalence of microrganisms in the aetiology of spontaneous abortion during the first trimester of pregnancy has not yet been well established. In this study, we analysed the tissues relative to the product of conception from abortions during the first trimester (51 spontaneous abortions and 56 voluntary pregnancy interruptions) in women attending the Gynecological Sciences Perinatology and Puericulture Department of "Policlinico Umberto I". Specimens were investigated by cultural methods for the presence of yeasts, gram positive, gram negative bacteria, and genital mycoplasma. By molecular diagnostic procedures, DNA sequences of Chlamydia trachomatis, herpes simplex viruses, adenovirus, human papillomaviruses and human polyomaviruses BK and JC were searched. None of these agents could be found in voluntary pregnancy interruption samples, with the exception of 3.6% of specimens positive for adenovirus, whereas spontaneous abortion tissues were positive for at least one microrganism by 31.5%. Data analysis showed the occurrence of both monomicrobial and polymicrobial infections.

Heterogeneity of virulence-related properties in Listeria monocytogenes strains isolated from patients with haematological malignancies.

Listeria monocytogenes is an intracellular foodborne pathogen of humans and animals for which there are indications of virulence differences among strains. Various virulence properties related to different phases of infection process were investigated in L. monocytogenes strains isolated from patients affected by haematological malignancies. In these isolates, besides to the clinical history, we analysed the haemolysin production, the survival to acidic pH, the ability to enter and proliferate in human intestinal-like and human macrophagic-like cells, as well as the allelic polymorphism of the actA gene involved intracellular movement. A general heterogeneity in the virulence properties was detected which did not appear correlated with the clinical outcome of listeriosis but more probably was influenced by the status of the immune defence of the host.

Detection of Listeria monocytogenes in Italian-style soft cheeses.

AIMS: A rapid detection system specific for Listeria monocytogenes and based on the polymerase chain reaction (PCR) was developed. METHODS AND RESULTS: Primers annealing to the coding region of the actA gene, critically involved in virulence and capable of discrimination between two different alleles naturally occurring in L. monocytogenes, have been utilized. The procedure was applied to recover L. monocytogenes cells in artificially contaminated fresh Italian soft cheeses (mozzarella, crescenza and ricotta). Low levels of L. monocytogenes were detected in mozzarella and crescenza homogenates (0.04-0.4 and 4 CFU g(-1), respectively) whereas in ricotta the detection limit was higher (40 CFU g(-1)). CONCLUSIONS: This PCR-based assay is highly specific as primers used recognize the DNA from different L. monocytogenes strains of clinical and food origin, while no amplification products result with any other Listeria spp. strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlighted a low-cost and rapid procedure that can be appropriated for the detection in real time of low L. monocytogenes levels in soft cheese.

The ABILHAND questionnaire as a measure of manual ability in chronic stroke patients: Rasch-based validation and relationship to upper limb impairment.

BACKGROUND AND PURPOSE: Chronic hemiparetic patients often retain the ability to manage activities requiring both hands, either through the use of the affected arm or compensation with the unaffected limb. A measure of this overall ability was developed by adapting and validating the ABILHAND questionnaire through the Rasch measurement model. ABILHAND measures the patient's perceived difficulty in performing everyday manual activities. METHODS: One hundred three chronic (>6 months) stroke outpatients (62% men; mean age, 63 years) were assessed (74 in Belgium, 29 in Italy). They lived at home and walked independently and were screened for the absence of major cognitive deficits (dementia, aphasia, hemineglect). The patients were administered the ABILHAND questionnaire, the Brunnström upper limb motricity test, the box-and-block manual dexterity test, the Semmes-Weinstein tactile sensation test, and the Geriatric Depression Scale. The brain lesion type and site were recorded. ABILHAND results were analyzed with the use of Winsteps Rasch software. RESULTS: The Rasch refinement of ABILHAND led to a change from the original unimanual and bimanual 56-item, 4-level scale to a bimanual 23-item, 3-level scale. The resulting ability scale had sufficient sensitivity to be clinically useful. Rasch reliability was 0.90, and the item-difficulty hierarchy was stable across demographic and clinical subgroups. Grip strength, motricity, dexterity, and depression were significantly correlated with the ABILHAND measures. CONCLUSIONS: The ABILHAND questionnaire results in a valid person-centered measure of manual ability in everyday activities. The stability of the item-difficulty hierarchy across different patient classes further supports the clinical application of the scale.

ABILHAND: a Rasch-built measure of manual ability.

OBJECTIVE: To apply the Rasch measurement model to the development of a clinical tool for measuring manual (dis)ability (ABILHAND). DESIGN: Manual ability was evaluated in terms of the difficulty perceived by a hand-impaired patient on 57 representative unimanual or bimanual activities. SETTING: A clinical laboratory. PATIENTS: Eighteen rheumatoid arthritis patients (14 women, 4 men) were interviewed after wrist arthrodesis (10 right, 4 left, and 4 both wrists). Their ages ranged from 38 to 77 years, time since diagnosis ranged from 7 to 41 years, and time since surgery ranged from 0.5 to 17 years. MAIN OUTCOME MEASURE: ABILHAND, administered at a mean duration of 7 years after arthrodesis. RESULTS: Forty-six of the 57 items define a common, single manual ability continuum with widespread measurement range and regular item distribution. Items relating to feeding, grooming, and dressing upper body worked consistently with their counterparts in other disability scales. More difficult items extend the measurement range beyond that of most existing manual ability scales. CONCLUSION: Even in a small sample of patients, using the Rasch methodology enabled the investigators to produce a useful scale of manual (dis)ability and to define manual ability as a unique construct, at least in patients with rheumatoid arthritis.

Anterior lumbar interbody fusion. A minimum 10-year follow-up.

STUDY DESIGN: One hundred eight patients from a consecutive series of 125 anterior lumbar interbody fusions were invited to take part in a clinical outcome assessment (including plain radiography and magnetic resonance imaging of the lumbosacral spine) more than 10 years after the original surgery. OBJECTIVES: By standardizing the reporting of outcome, to determine whether the duration of patient follow-up influences the outcome of surgery, with particular reference to the effects of compensation and psychological status. SUMMARY OF BACKGROUND DATA: The success rates of lumbar spinal fusion surgery reported in the literature vary widely. The lack of standardization of measures of patient outcome limits the value of study comparisons. Evaluation of the efficacy of spinal fusion is further compounded by the adverse effects of both compensation and psychological disturbance on the reporting of outcome. METHODS: One hundred three patients agreed to take part in a clinical outcome assessment by completing a comprehensive low-back questionnaire that included demographic, compensation, and employment details. Eighty-seven of these cases also agreed to undergo radiographic evaluation and magnetic resonance imaging of the lumbar spine. Subjective assessment of outcome was based on a 10-point analog pain scale as well as patient opinion regarding the success of surgery. A more objective assessment of outcome was made using the Low-Back Outcome Score. Psychological status was determined by combining the Modified Somatic Perception Questionnaire and the Zung Depression Scale. The effects of radiologic fusion, compensation status, psychological status, and reoperation on the various outcome measures were assessed and compared with the results reported in a separate but similar series of patients with a minimum follow-up of 2 years. RESULTS: Seventy-eight percent of patients rated themselves as having "complete relief" or "a good deal of relief," but only 34% fell into the "excellent" or "good" category using the Low-Back Outcome Score. The clinical outcome was not associated with the presence of radiologic fusion and was not influenced by the compensation status. Psychological disturbance at review and reoperation, however, did influence the reporting of outcome and were significantly correlated with the Low-Back Outcome Score. With the exception of the effects of compensation, these results were remarkably similar to the findings in the 2-year study. CONCLUSIONS: The findings of the study suggest that the assessment of outcome of lumbar interbody fusion is strongly compounded by the psychological make-up of the patient and that this effect is maintained in the long term. However, the negative effect of compensation observed at 2 years seems to dissipate with time and becomes insignificant at 10 years.

Short-term effect of hand-arm vibration exposure on tactile sensitivity and manual skill.

OBJECTIVES: The present study investigated whether the impairment of tactile sensitivity after exposure to vibration disturbs the motor control of precision handling and, if so, whether it can result in an increased risk of injury during or after tasks involving the use of vibrating tools. METHODS: Twelve men were manually exposed to vibration from an electric sander for 30 min. Cutaneous sensitivity was quantified to measuring the pressure perception threshold and vibration perception threshold (125 Hz) on the pulp of the second finger. Manipulative skill was evaluated by grip-lift movements and the Purdue pegboard test. RESULTS: The vibration perception threshold increased very significantly from 94.0 dB (0.06 m/s2) before the vibration exposure to 127.5 dB (3.2 m/s2) immediately after the exposure. The pressure perception threshold tended to increase after vibration exposure, although not significantly, but manipulative skill was not altered. CONCLUSIONS: Change in vibration perception threshold was not associated with a significant increase in the pressure perception threshold or a perturbation of manual skill. Therefore, in conditions similar to those of our experiment, the safety of a precision task does not appear to be reduced after such vibration exposure.

Bilateral synovitis in symptomatic unilateral transient synovitis of the hip: an ultrasonographic study in 56 children.

56 children with a clinical diagnosis of unilateral transient synovitis of the hip underwent bilateral sonographic assessment. On the anterior scan, the distance between the femoral neck and the fibrous joint capsule was measured. This distance, which we call the synovial capsular complex distance, was compared with age-dependent normal values. An increased distance was found in all 56 symptomatic hips (mean 10 mm, SD 1.8). This distance was also increased in 14 hips on the contralateral side (mean 8 mm, SD 1.6). An effusion was demonstrated in 53 symptomatic hips and in 8 hips on the contralateral side. These findings indicate that in one quarter of children with symptoms of unilateral transient synovitis the contralateral hip may have an increased synovial capsular complex distance due to synovial swelling or joint effusion, suggesting an asymptomatic synovitis. We therefore recommend a comparison of the synovial capsular complex distance on the symptomatic side with age-related normal values, in addition to a comparison with the asymptomatic hip.

Energy-saving gait mechanics with head-supported loads.

In many areas of the world that lack a transportation infrastructure, people routinely carry extraordinary loads supported by their heads, for example the Sherpa of the Himalayas and the women of East Africa. It has previously been shown that African women from the Kikuyu and Luo tribes can carry loads substantially more cheaply than army recruits; however, the mechanism for their economy has remained unknown. Here we investigate, using a force platform, the mechanics of carrying head-supported loads by Kikuyu and Luo women. The weight-specific mechanical work, required to maintain the motion of the common centre of mass of the body and load, decreases with load in the African women, whereas it increases in control subjects. The decrease in work by the African women is a result of a greater conservation of mechanical energy resulting from an improved pendulum-like transfer of energy during each step, back and forth between gravitational potential energy and kinetic energy of the centre of mass.

Magnetic resonance imaging assessment of disc degeneration 10 years after anterior lumbar interbody fusion.

STUDY DESIGN: One-hundred-eight patients from a consecutive series of 125 anterior lumbar interbody fusions were invited to participate in a radiographic and magnetic resonance imaging assessment more than 10 years after the original surgery. OBJECTIVE: The aim of this study was to determine the long-term incidence of disc degeneration adjacent to an anterior interbody fusion and to determine whether this was influenced by the length of the fusion. SUMMARY OF BACKGROUND DATA: Biomechanical studies in human cadaver spines support the view that fusion in the lumbar spine is likely to be associated with an increased incidence of degeneration at adjacent levels, but there is little reliable information on the incidence of juxtafusion degeneration from the few long-term studies of lumbar spine fusion reported in the literature. METHODS: Eighty-seven patients agreed to take part in the study, but in six cases the magnetic resonance imaging procedure had to be abandoned. Of the 81 patients who underwent both radiographic assessment and magnetic resonance imaging scanning, preoperative discography had demonstrated a normal disc above the level of the fusion in 52. This group of patients formed the basis of this report. Each sequence of magnetic resonance imaging films was examined by one radiologist. The signal strength was assessed on T2-weighted images in the sagittal plane and disc morphology was assessed on T1- and T2-weighted sagittal and T1-weighted axial images. Fusion status was assessed on the plain films and magnetic resonance imaging. RESULTS: The incidence of a normal adjacent intervertebral disc in cases of solid fusions to the sacrum was 68%. This was not influenced by the length of the fusion. CONCLUSION: The findings of this study suggest that degeneration after an anterior lumbar interbody fusion is determined more by individual characteristics than by the fusion itself.