Functional outcome assessment of lower limb amputees and prosthetic users with a 2-minute walk test

The aim of this study was to assess the functional outcome of a population of lower limb amputees supplied with prosthesis. The research was conducted from June to September of 2010 at the Center for Physical and Rehabilitation Medicine, of the Clinical Hospital Center Rijeka, Croatia. The study included 50 adult subjects of both genders with a unilateral transtibial or transfemoral lower limb amputations. The 2-minute walk test (2MWT) was used to assess the functional outcome of these individuals. Data were statistically analyzed. Subjects were divided into groups according to age. The best results were obtained by subjects between the age of 45 and 59 years. The difference between groups was statistically significant (p < 0.001). Taking into account the cause of amputation, there was a statistically significant difference in the results of the 2 MWT between subjects in whom the cause of amputation was circulatory and those where the cause of the amputation was not due to circulatory problems. The best results were obtained in subjects in whom the cause of amputation was not circulatory (p = 0.009). Considering the level of amputation there was a statistically significant difference in the results of the 2MWT between subjects with transtibial and those with transfemoral amputations. Better results were obtained in transtibial amputees (p = 0.039). Considering the first prosthetic supply, better results were obtained in subjects using prosthetic devices over 9 years (p = 0.031). Our research confirmed that age, gender, level and cause of amputation, including the time from the first prosthetic supply have an effect on the 2MWT results.     

Radial extracorporeal shock wave therapy in the treatment of shoulder calcific tendinitis

Shoulder calcific lesions of the rotator cuff are a common problem in physiatric and orthopedic practice. The lesions are mostly located in the supraspinatus tendon, close to the insertion area in the critical zone. Patients are usually treated conservatively by nonsteroid antiinflammatory drugs, analgesic drugs, local injections, physiotherapy and rarely by applying surgical procedures. Painful shoulder gives rise to functional disabilities and may sometimes lead to pharmacological overuse. In the last twenty years, extracorporeal shock wave therapy (ESWT) has been frequently used in the treatment of calcific tendinopathies. We have evaluated the effectiveness of radial ESWT on the group of 30 patients, aged between 28 and 58 years, with calcific tendinitis of the shoulder. Criotherapy, medical exercises and radial ESWT were applied. We used the radial ESWT device (BTL-5000 SWT, Columbia, USA), 3 bars pressure, 10 Hz frequency, 2000 shocks. Patients were examined before the beginning of the treatment, immediately after the treatment, and 6 months later. The treatment included measurement of the range of motion (ROM), measurement of voluntary isometric contraction of shoulder muscles with manual muscle test (MMT), and subjective assessment of pain intensity with visual analogue scale (VAS). X-ray was done before and 6 month after treatment. The study has shown the efficiency of the treatment with radial ESWT. The level of statistical significance was determined with student t-test. Radial ESWT applied to patients with shoulder calcific lesions of the rotator cuff resulted in pain relief increase in the range of motion and increase in the muscular strength. As shown by X-ray, these results were followed by the decrease in the size of the rotator cuff calcifications.     

"Brilliant Blue G" and "Membrane Blue Dual" assisted vitrectomy for macular hole

The aim of this study is to evaluate vital dyes "Brilliant Blue G" (BBG) and "Membrane Blue Dual" (MBD) for intraoperative staining of the inner limiting membrane (ILM) during vitrectomy for macular hole (MH). Retrospective, comparative case series on 18 eyes with macular holes who underwent "23 and 25 gauge" pars plana vitrectomy. Main outcome measurements were staining intensity and characteristics, visual acuity, visual field, OCT measurements and complications over a period of 6 months. With the help of BBG and MBD successfully was removed complete ILM in 14 eyes. Postoperative visual acuity was improved in 12 patients, unchanged in 2 patients and worse in 4 patients. Central retinal thickness showed significant postoperative reduction with closure of macular hole. OCT values range were from -10 to -250 microm. No visual field defects and no adverse effects were found. BBG and MBD successfully identificate internal limiting membrane during vitrectomy for MH. Good anatomical and functional results are achieved with the use of both vital dyes.     

Evidence for 1,8-dihydroxynaphthalene melanin in three halophilic black yeasts grown under saline and non-saline conditions

The ascomycetous black yeasts Hortaea werneckii, Phaeotheca triangularis, and Trimmatostroma salinum are halophilic fungi that inhabit hypersaline water of solar salterns. They are characterized by slow, meristematic growth and very thick, darkly pigmented cell walls. The dark pigment, generally thought to be melanin, is consistently present in their cell walls when they grow under saline and non-saline conditions. We used the inhibitor tricyclazole to test the fungi in this study for the presence of 1,8-dihydroxynaphthalene (DHN)-melanin biosynthesis, since fungal melanins reportedly are derived either from DHN, tyrosine via 3,4-dihydroxyphenylalanine, gamma-glutaminyl-3,4-dihydroxybenzene, or catechol. Tricyclazole-treated cultures of the fungi were reddish-brown in color and contained typical intermediates of the DHN-melanin pathway, as demonstrated by high-performance liquid chromatography. This investigation showed that the three fungi synthesized DHN-melanin under saline and non-saline growth conditions.     

Characterizing the trophic niches of stocked and resident cyprinid fishes: consistency in partitioning over time,space and body sizes

Hatchery‐reared fish are commonly stocked into freshwaters to enhance recreational angling. As these fishes are often of high trophic position and attain relatively large sizes, they potentially interact with functionally similar resident fishes and modify food‐web structure. Hatchery‐reared barbel Barbus barbus are frequently stocked to enhance riverine cyprinid fish communities in Europe; these fish can survive for over 20 years and exceed 8 kg. Here, their trophic consequences for resident fish communities were tested using cohabitation studies, mainly involving chub Squalius cephalus, a similarly large‐bodied, omnivorous and long‐lived species. These studies were completed over three spatial scales: pond mesocosms, two streams and three lowland rivers, and used stable isotope analysis. Experiments in mesocosms over 100 days revealed rapid formation of dietary specializations and discrete trophic niches in juvenile B. barbus and S. cephalus. This niche partitioning between the species was also apparent in the streams over 2 years. In the lowland rivers, where fish were mature individuals within established populations, this pattern was also generally apparent in fishes of much larger body sizes. Thus, the stocking of these hatchery‐reared fish only incurred minor consequences for the trophic ecology of resident fish, with strong patterns of trophic niche partitioning and diet specialization. Application of these results to decision‐making frameworks should enable managers to make objective decisions on whether cyprinid fish should be stocked into lowland rivers according to ecological risk.     

New inhibitors of fungal 17beta-hydroxysteroid dehydrogenase based on the [1,5]-benzodiazepine scaffold

The synthesis and activity of a new series of non-steroidal inhibitors of 17beta-hydroxysteroid dehydrogenase that are based on a 1,5-benzodiazepine scaffold are presented. Their inhibitory potential was screened against 17beta-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus (17beta-HSDcl), a model enzyme of the short-chain dehydrogenase/reductase superfamily. Some of these compounds are potent inhibitors of 17beta-HSDcl activity, with IC50 values in the low micromolar range and represent promising lead compounds that should be further developed and investigated as inhibitors of human 17beta-HSD isoforms, which are the enzymes associated with the development of many hormone-dependent and neuronal diseases.     

Evaluation of the SELDI-TOF MS technique for protein profiling of pancreatic islets exposed to glucose and oleate

The aim of the study was to evaluate the SELDI-TOF MS technique for pancreatic islet research. Mouse islets were cultured at low or high glucose levels in the absence or presence of oleate and characterized by measuring insulin secretion and oxygen tension. Subsequently, the islets were protein profiled. Up to 200 different peaks could be detected in a single experiment with the majority of peaks corresponding to proteins with masses below 30 kDa. By combining different protein arrays, the number of detected peaks could be increased further. The optimal binding of islet proteins was achieved using the anionic exchange array and phosphate buffer (pH 6) when the binding of insulin was low, which allowed other less abundant proteins to be captured. When islets from different culture conditions were profiled and analyzed, in total 25 proteins were found to be oleate/glucose-regulated. An oleate-regulated protein was chosen for identification work, which was conducted by passive elution from SDS-PAGE gels and subsequent in-gel trypsin digestion and MALDI-TOF MS. The protein was identified as peptidyl-prolyl isomerase B (PPI-B). In conclusion, the study demonstrates that SELDI-technique can be used not only to obtain islet protein patterns but is also helpful in the subsequent identification of differentially expressed proteins.     

A hypoallergenic vaccine obtained by tail-to-head restructuring of timothy grass pollen profilin, Phl p 12, for the treatment of cross-sensitization to profilin

Profilins are highly cross-reactive allergens in pollens and plant food. In a paradigmatic approach, the cDNA coding for timothy grass pollen profilin, Phl p 12, was used as a template to develop a new strategy for engineering an allergy vaccine with low IgE reactivity. Non-IgE-reactive fragments of Phl p 12 were identified by synthetic peptide chemistry and restructured (rs) as a new molecule, Phl p 12-rs. It comprised the C terminus of Phl p 12 at its N terminus and the Phl p 12 N terminus at its C terminus. Phl p 12-rs was expressed in Escherichia coli and purified to homogeneity. Determination of secondary structure by circular dichroism indicated that the restructuring process had reduced the IgE-reactive alpha-helical contents of the protein but retained its beta-sheet conformation. Phl p 12-rs exhibited reduced IgE binding capacity and allergenic activity but preserved T cell reactivity in allergic patients. IgG Abs induced by immunization of mice and rabbits with Phl p 12-rs cross-reacted with pollen and food-derived profilins. Recombinant Phl p 12-rs, rPhl p 12-rs, induced less reaginic IgE to the wild-type allergen than rPhl p 12. However, the rPhl p 12-rs-induced IgGs inhibited allergic patients' IgE Ab binding to profilins to a similar degree as those induced by immunization with the wild type. Phl p 12-rs specific IgG inhibited profilin-induced basophil degranulation. In conclusion, a restructured recombinant vaccine was developed for the treatment of profilin-allergic patients. The strategy of tail-to-head reassembly of hypoallergenic allergen fragments within one molecule represents a generally applicable strategy for the generation of allergy vaccines.     

Lifestyle-Related Factors and Atopy in Seven Danish Population-Based Studies from Different Time Periods

Background

The prevalence of allergic respiratory disease tends to increase in populations that adopt the so-called Westernized lifestyle. We investigated the association between atopy and several possible lifestyle-related factors in seven Danish population-based studies.

Methods

A total of 20048 persons participated in the seven studies. We used logistic regression to analyse the associations between possible determinants and atopy defined as serum specific IgE or skin prick test positivity against inhalant allergens. Associations were expressed as odds ratios (ORs) with 95% confidence intervals (95% CIs). In addition, individual participant data meta-analyses were performed.

Results

Atopy was significantly associated with younger age (OR per 1 year increase in age: 0.97; 95% CI: 0.97, 0.98); male sex (OR for males versus females: 1.34; 95% CI: 1.24, 1.45), heavy drinking (OR for heavy drinkers versus light drinkers: 1.15; 95% CI: 1.04, 1.27), never smoking (OR for current versus never smokers: 0.73; 95% CI: 0.67, 0.80), and higher educational level (OR for educated versus uneducated: 1.27; 95% CI: 1.15, 1.41). Atopy was not associated with blood pressure, serum total cholesterol, physical activity or body mass except in women only, where we found a positive association (OR for obese vs. normal weight: 1.18; 95% CI: 1.00, 1.39) with p_trend = 0.032.

Conclusions

Of interest for preventive purposes, we found that atopy was associated with some of the reversible lifestyle-related factors that characterize a Westernized lifestyle.     

Mycena species can be opportunist-generalist plant root invaders

Traditional strict separation of fungi into ecological niches as mutualist, parasite or saprotroph is increasingly called into question. Sequences of assumed saprotrophs have been amplified from plant root interiors, and several saprotrophic genera can invade and interact with host plants in laboratory growth experiments. However, it is uncertain if root invasion by saprotrophic fungi is a widespread phenomenon and if laboratory interactions mirror field conditions. Here, we focused on the widespread and speciose saprotrophic genus Mycena and performed (1) a systematic survey of their occurrences (in ITS1/ITS2 datasets) in mycorrhizal roots of 10 plant species, and (2) an analysis of natural abundances of ¹³C/¹⁵N stable isotope signatures of Mycena basidiocarps from five field locations to examine their trophic status. We found that Mycena was the only saprotrophic genus consistently found in 9 out of 10 plant host roots, with no indication that the host roots were senescent or otherwise vulnerable. Furthermore, Mycena basidiocarps displayed isotopic signatures consistent with published ¹³C/¹⁵N profiles of both saprotrophic and mutualistic lifestyles, supporting earlier laboratory-based studies. We argue that Mycena are widespread latent invaders of healthy plant roots and that Mycena species may form a spectrum of interactions besides saprotrophy also in the field.