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Receptor activity-modifying proteins (RAMPs) form complexes with G protein-coupled receptors (GPCRs) and may regulate their cellular trafficking and pharmacology. RAMP interactions have been identified for about 50 GPCRs, but only a few GPCR-RAMP complexes have been studied in detail. To elucidate a comprehensive GPCR-RAMP interactome, we created a library of 215 dual epitope-tagged (DuET) GPCRs representing all GPCR subfamilies and coexpressed each GPCR with each of the three RAMPs. Screening the GPCR-RAMP pairs with customized multiplexed suspension bead array (SBA) immunoassays, we identified 122 GPCRs that showed strong evidence for interaction with at least one RAMP. We screened for interactions in three cell lines and found 23 endogenously expressed GPCRs that formed complexes with RAMPs. Mapping the GPCR-RAMP interactome expands the current system-wide functional characterization of RAMP-interacting GPCRs to inform the design of selective therapeutics targeting GPCR-RAMP complexes.

Background: Self-sampling of dried blood spots (DBS) offers new routes to gather valuable health-related information from the general population. Yet, the utility of using deep proteome profiling from home-sampled DBS to obtain clinically relevant insights about SARS-CoV-2 infections remains largely unexplored., Methods: Our study involved 228 individuals from the general Swedish population who used a volumetric DBS sampling device and completed questionnaires at home during spring 2020 and summer 2021. Using multi-analyte COVID-19 serology, we stratified the donors by their response phenotypes, divided them into three study sets, and analyzed 276 proteins by proximity extension assays (PEA). After normalizing the data to account for variances in layman-collected samples, we investigated the association of DBS proteomes with serology and self-reported information., Results: Our three studies display highly consistent variance of protein levels and share associations of proteins with sex (e.g., MMP3) and age (e.g., GDF-15). Studying seropositive (IgG + ) and seronegative (IgG - ) donors from the first pandemic wave reveals a network of proteins reflecting immunity, inflammation, coagulation, and stress response. A comparison of the early-infection phase (IgM + IgG - ) with the post-infection phase (IgM - IgG + ) indicates several proteins from the respiratory system. In DBS from the later pandemic wave, we find that levels of a virus receptor on B-cells differ between seropositive (IgG + ) and seronegative (IgG - ) donors., Conclusions: Proteome analysis of volumetric self-sampled DBS facilitates precise analysis of clinically relevant proteins, including those secreted into the circulation or found on blood cells, augmenting previous COVID-19 reports with clinical blood collections. Our population surveys support the usefulness of DBS, underscoring the role of timing the sample collection to complement clinical and precision health monitoring initiatives., (© 2024. The Author(s).)

G protein-coupled receptors (GPCRs) control critical cellular signaling pathways. Therapeutic agents including anti-GPCR antibodies (Abs) are being developed to modulate GPCR function. However, validating the selectivity of anti-GPCR Abs is challenging because of sequence similarities among individual receptors within GPCR subfamilies. To address this challenge, we developed a multiplexed immunoassay to test >400 anti-GPCR Abs from the Human Protein Atlas targeting a customized library of 215 expressed and solubilized GPCRs representing all GPCR subfamilies. We found that ~61% of Abs tested were selective for their intended target, ~11% bound off-target, and ~28% did not bind to any GPCR. Antigens of on-target Abs were, on average, significantly longer, more disordered, and less likely to be buried in the interior of the GPCR protein than the other Abs. These results provide important insights into the immunogenicity of GPCR epitopes and form a basis for designing therapeutic Abs and for detecting pathological auto-Abs against GPCRs.

Cholestatic itch is a severe and debilitating symptom in liver diseases with limited treatment options. The class A G protein-coupled receptor (GPCR) Mas-related GPCR subtype X4 (MRGPRX4) has been identified as a receptor for bile acids, which are potential cholestatic pruritogens. An increasing number of GPCRs have been shown to interact with receptor activity-modifying proteins (RAMPs), which can modulate different aspects of GPCR biology. Using a combination of multiplexed immunoassay and proximity ligation assay, we show that MRGPRX4 interacts with RAMPs. The interaction of MRGPRX4 with RAMP2, but not RAMP1 or 3, causes attenuation of basal and agonist-dependent signaling, which correlates with a decrease of MRGPRX4 cell surface expression as measured using a quantitative NanoBRET pulse-chase assay. Finally, we use AlphaFold Multimer to predict the structure of the MRGPRX4-RAMP2 complex. The discovery that RAMP2 regulates MRGPRX4 may have direct implications for future drug development for cholestatic itch., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

We previously demonstrated that we could hijack the fungal pheromone signaling pathway to provide a living yeast biosensor where peptide biomarkers were recognized by G-protein-coupled receptors and engineered to transcribe a readout. Here, we demonstrated that the protease could be reintroduced to the biosensor to provide a simple mechanism for distinguishing single-amino-acid changes in peptide ligands that, otherwise, would likely be difficult to detect using binding-based assays. We characterized the dose-response curves for five fungal pheromone G-protein-coupled receptors, peptides, and proteases -Saccharomyces cerevisiae , Candida albicans , Schizosaccharomyces pombe , Schizosaccharomyces octosporus , and Schizosaccharomyces japonicus . Alanine scanning was carried out for the most selective of these- S. cerevisiae and C. albicans -with and without the protease. Two peptide variants were discovered, which showed diminished cleavage by the protease (CaPep2A and CaPep2A13A). Those peptides were then distinguished by utilizing the biosensor strains with and without the protease, which selectively cleaved and altered the apparent concentration of peptide required for half-maximal activation for 2 peptides-CaPep and CaPep13A, respectively-by more than one order of magnitude. These results support the hypothesis that the living yeast biosensor with a sequence-specific protease can translate single-amino-acid changes into more than one order of magnitude apparent shift in the concentration of peptide required for half-maximal activation. With further engineering by computational modeling and directed evolution, the biosensor could likely distinguish a wide variety of peptide sequences beyond the alanine scanning carried out here. In the future, we envision incorporating proteases into our living yeast biosensor for use as a point of care diagnostic, a scalable communication language, and other applications., Competing Interests: Competing interests: A provisional patent application no. 63/228,577 was filed 2021 Aug naming V.W.C., T.C., and M. Fantini (Columbia University, USA) as inventors and assigned to The Trustees of Columbia University in the City of New York. V.W.C. holds equity and is Chairman of the Board and President of Celsyntec, Inc. All other authors declare no competing interests.

Serological testing is essential to curb the consequences of the COVID-19 pandemic. However, most assays are still limited to single analytes and samples collected within healthcare. Thus, we establish a multianalyte and multiplexed approach to reliably profile IgG and IgM levels against several versions of SARS-CoV-2 proteins (S, RBD, N) in home-sampled dried blood spots (DBS). We analyse DBS collected during spring of 2020 from 878 random and undiagnosed individuals from the population in Stockholm, Sweden, and use classification approaches to estimate an accumulated seroprevalence of 12.5% (95% CI: 10.3%-14.7%). This includes 5.4% of the samples being IgG + IgM + against several SARS-CoV-2 proteins, as well as 2.1% being IgG - IgM + and 5.0% being IgG + IgM - for the virus' S protein. Subjects classified as IgG + for several SARS-CoV-2 proteins report influenza-like symptoms more frequently than those being IgG + for only the S protein (OR = 6.1; p < 0.001). Among all seropositive cases, 30% are asymptomatic. Our strategy enables an accurate individual-level and multiplexed assessment of antibodies in home-sampled blood, assisting our understanding about the undiagnosed seroprevalence and diversity of the immune response against the coronavirus.

Metallic nanoparticles are an important and widely used materials in development of nano-enabled medicine. For that reason, their interaction with biological molecules has to be systematically examined, as use of nanoparticles can lead to altered biological functions. In this study, we evaluated the interaction between silver nanoparticles (AgNPs) and two important plasma transport proteins - albumin and α-1-acid glycoprotein. To investigate comprehensively how different physico-chemical properties impact interaction of proteins with nanosurface, AgNPs of different size, shape and surface coating was prepared. The study was conducted using UV-Vis absorption, fluorescence, inductively coupled plasma mass spectrometry, circular dichroism spectroscopy, transmission electron microscopy, dynamic and electrophoretic light scattering techniques. The results showed significant complexities of the nano-bio interface and binding affinities of proteins onto surface of different AgNPs, which were affected by both AgNPs and protein properties. The most significant role on AgNPs-protein interaction had the coating agents used for AgNPs surface stabilization. Our findings should improve safe-by-design approach to development of the metallic nanomaterials for medical use., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Protein biomarkers in biological fluids represent an important resource for improving the clinical management of diseases. Current proteomics technologies are capable of performing high-throughput and multiplex profiling in different types of fluids, often leading to the shortlisting of tens of candidate biomarkers per study. However, before reaching any clinical setting, these discoveries require thorough validation and an assay that would be suitable for routine analyses. In the path from biomarker discovery to validation, the performance of the assay implemented for the intended protein quantification is extremely critical toward achieving reliable and reproducible results. Development of robust sandwich immunoassays for individual candidates is challenging and labor and resource intensive, and multiplies when evaluating a panel of interesting candidates at the same time. Here we describe a versatile pipeline that facilitates the systematic and parallel development of multiple sandwich immunoassays using a bead-based technology.

An important aspect of precision medicine is to probe the stability in molecular profiles among healthy individuals over time. Here, we sample a longitudinal wellness cohort with 100 healthy individuals and analyze blood molecular profiles including proteomics, transcriptomics, lipidomics, metabolomics, autoantibodies and immune cell profiling, complemented with gut microbiota composition and routine clinical chemistry. Overall, our results show high variation between individuals across different molecular readouts, while the intra-individual baseline variation is low. The analyses show that each individual has a unique and stable plasma protein profile throughout the study period and that many individuals also show distinct profiles with regards to the other omics datasets, with strong underlying connections between the blood proteome and the clinical chemistry parameters. In conclusion, the results support an individual-based definition of health and show that comprehensive omics profiling in a longitudinal manner is a path forward for precision medicine.

Despite recognizing aging as a common risk factor of many human diseases, little is known about its molecular traits. To identify age-associated proteins circulating in human blood, we screened 156 individuals aged 50-92 using exploratory and multiplexed affinity proteomics assays. Profiling eight additional study sets (N = 3,987), performing antibody validation, and conducting a meta-analysis revealed a consistent age association ( P = 6.61 × 10 -6 ) for circulating histidine-rich glycoprotein (HRG). Sequence variants of HRG influenced how the protein was recognized in the immunoassays. Indeed, only the HRG profiles affected by rs9898 were associated with age and predicted the risk of mortality (HR = 1.25 per SD; 95% CI = 1.12-1.39; P = 6.45 × 10 -5 ) during a follow-up period of 8.5 yr after blood sampling (IQR = 7.7-9.3 yr). Our affinity proteomics analysis found associations between the particular molecular traits of circulating HRG with age and all-cause mortality. The distinct profiles of this multipurpose protein could serve as an accessible and informative indicator of the physiological processes related to biological aging., (© 2020 Hong et al.)

Background: Precision medicine approaches aim to tackle diseases on an individual level through molecular profiling. Despite the growing knowledge about diseases and the reported diversity of molecular phenotypes, the descriptions of human health on an individual level have been far less elaborate., Methods: To provide insights into the longitudinal protein signatures of well-being, we profiled blood plasma collected over one year from 101 clinically healthy individuals using multiplexed antibody assays. After applying an antibody validation scheme, we utilized > 700 protein profiles for in-depth analyses of the individuals' short-term health trajectories., Findings: We found signatures of circulating proteomes to be highly individual-specific. Considering technical and longitudinal variability, we observed that 49% of the protein profiles were stable over one year. We also identified eight networks of proteins in which 11-242 proteins covaried over time. For each participant, there were unique protein profiles of which some could be explained by associations to genetic variants., Interpretation: This observational and non-interventional study identifyed noticeable diversity among clinically healthy subjects, and facets of individual-specific signatures emerged by monitoring the variability of the circulating proteomes over time. To enable more personal hence precise assessments of health states, longitudinal profiling of circulating proteomes can provide a valuable component for precision medicine approaches., Funding: This work was supported by the Erling Persson Foundation, the Swedish Heart and Lung Foundation, the Knut and Alice Wallenberg Foundation, Science for Life Laboratory, and the Swedish Research Council., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Receptor activity-modifying proteins (RAMPs) have been shown to modulate the functions of several G protein-coupled receptors (GPCRs), but potential direct interactions among the three known RAMPs and hundreds of GPCRs have never been investigated. Focusing mainly on the secretin-like family of GPCRs, we engineered epitope-tagged GPCRs and RAMPs, and developed a multiplexed suspension bead array (SBA) immunoassay to detect GPCR-RAMP complexes from detergent-solubilized lysates. Using 64 antibodies raised against the native proteins and 4 antibodies targeting the epitope tags, we mapped the interactions among 23 GPCRs and 3 RAMPs. We validated nearly all previously reported secretin-like GPCR-RAMP interactions, and also found previously unidentified RAMP interactions with additional secretin-like GPCRs, chemokine receptors, and orphan receptors. The results provide a complete interactome of secretin-like GPCRs with RAMPs. The SBA strategy will be useful to search for additional GPCR-RAMP complexes and other interacting membrane protein pairs in cell lines and tissues.

The plasma proteome offers a clinically useful window into human health. Recent advances from highly multiplexed assays now call for appropriate pipelines to validate individual candidates. Here, a workflow is developed to build dual binder sandwich immunoassays (SIA) and for proteins predicted to be secreted into plasma. Utilizing suspension bead arrays, ≈1800 unique antibody pairs are first screened against 209 proteins with recombinant proteins as well as EDTA plasma. Employing 624 unique antibodies, dilution-dependent curves in plasma and concentration-dependent curves of full-length proteins for 102 (49%) of the targets are obtained. For 22 protein assays, the longitudinal, interindividual, and technical performance is determined in a set of plasma samples collected from 18 healthy subjects every third month over 1 year. Finally, 14 of these assays are compared with with SIAs composed of other binders, proximity extension assays, and affinity-free targeted mass spectrometry. The workflow provides a multiplexed approach to screen for SIA pairs that suggests using at least three antibodies per target. This design is applicable for a wider range of targets of the plasma proteome, and the assays can be applied for discovery but also to validate emerging candidates derived from other platforms., (© 2019 KTH - Royal Institute of Technology. Proteomics Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objectives: Cancer-testis antigens (CTAs) are defined as proteins that are specifically expressed in testis or placenta and their expression is frequently activated in cancer. Due to their ability to induce an immune response, CTAs may serve as suitable targets for immunotherapy. The aim of this study was to evaluate if there is reactivity against CTAs in the plasma of non-small cell lung cancer (NSCLC) patients through the detection of circulating antibodies., Materials and Methods: To comprehensively analyze autoantibodies against CTAs the multiplexing capacities of suspension bead array technology was used. Bead arrays were created with 120 protein fragments, representing 112 CTAs. Reactivity profiles were measured in plasma samples from 133 NSCLC patients and 57 cases with benign lung diseases., Results: Altogether reactivity against 69 antigens, representing 81 CTAs, was demonstrated in at least one of the analyzed samples. Twenty-nine of the antigens (45 CTAs) demonstrated exclusive reactivity in NSCLC samples. Reactivity against cancer-testis antigen family 47; member A (CT47A) genes, P antigen family member 3 (PAGE3), variable charge X-linked (VCX), melanoma antigen family B1 (MAGEB1), lin-28 homolog B (LIN28B) and chromosome 12 open reading frame 54 (C12orf54) were only found in NSCLC patients at a frequency of 1%-4%. The presence of autoantibodies towards these six antigens was confirmed in an independent group of 34 NSCLC patients., Conclusion: We identified autoantibodies against CTAs in the plasma of lung cancer patients. The reactivity pattern of autoantibodies was higher in cancer patients compared to the benign group, stable over time, but low in frequency of occurrence. The findings suggest that some CTAs are immunogenic and that these properties can be utilized as immune targets., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The concept of antibody microarrays is one of the most versatile approaches within multiplexed immunoassay technologies. These types of arrays have increasingly become an attractive tool for the exploratory detection and study of protein abundance, function, pathways, and potential drug targets. Due to the properties of the antibody microarrays and their potential use in basic research and clinical analytics, various types of antibody microarrays have already been developed. In spite of the growing number of studies utilizing this technique, few reviews about antibody microarray technology have been presented to reflect the quality and future uses of the generated data. In this review, we provide a summary of the recent applications of antibody microarray techniques in basic biology and clinical studies, providing insights into the current trends and future of protein analysis.

Aim To evaluate biomechanical and clinical outcomes of a newly developed modification of the Wilson's osteotomy for hallux valgus: a three-dimensional subcaptial correction of the metatarsal head position with a simultaneous lateral and plantar shift with derotation intended to reduce displacement of the distal fragment and shortening of the first metatarsal bone. Methods Thirty four feet (28 female patients) underwent the new procedure and were evaluated before and 12 to 84 months (median=25.5) after the surgery. Results Plantar shift of the distal fragment was achieved in all feet. Shortening of the first metatarsal was moderate: ≤6 mm in 32/34 feet, 7 and 10 mm in the remaining two. Median difference in metatarsal index post- vs. pretreatment was -4.0. The hallux valgus angle, intermetatarsal and distal metatarsal articular angles were reduced in all feet. The American Orthopaedic Foot and Ankle Society score improved in all feet (median increase= 51.5). Conclusion The method allows for a lateral and plantar shift with derotation of the distal fragment and a mild/moderate shortening of the first metatarsal bone., (Copyright© by the Medical Assotiation of Zenica-Doboj Canton.)

Aim To identify modes of venous thromboembolism (VTE) prophylaxis in patients undergoing elective major orthopaedic surgery (total hip or knee arthroplasty, THA/TKA) at a single university-associated hospital in Croatia. Methods A retrospective analysis of consecutive patients subjected to THA or TKA over a two-year period (2014-2015) with a focus on anticoagulation during the first 15 post-surgical days (period of highest VTE risk). Results Of 603 identified patients three (0.5%) were not anticoagulated (haemophilia) and others received perioperative doses of low molecular weight heparins (LMWH). Overall, 228 (37.8%) patients received prophylaxis not involving warfarin, and 372 continued with short-term LMWH with switching to warfarin. They contributed a total of 1218 international normalized ratio (INR) values (median=3, range=1-8). These were consistently below the target INR range across the observed period. Between post-surgical days 6 and 15 (after the initial titration), 438 values were taken in patients treated with LMWH+warfarin and 92.7% were below, and only 6.8% within the target range; 580 values were taken in patients already switched to warfarin, 74% were below and only 25% within the range. Conclusion The prevailing mode of VTE prophylaxis was in a clear contrast to (then) actual professional guidelines, with inadequate monitoring and poor anticoagulation. There is no reason to expect a substantially different situation at other institutions across the country. The prevailing practice of VTE prophylaxis in major orthopaedic surgery in Croatia should be promptly abandoned and up-dated in agreement with the current state of the art., (Copyright© by the Medical Assotiation of Zenica-Doboj Canton.)

In-depth exploration and characterization of human serum and plasma proteomes is an attractive strategy for the identification of potential prognostic or diagnostic biomarkers. The possibility of analyzing larger numbers of samples in a high-throughput fashion has markedly increased with affinity-based microarrays, thus providing higher statistical power to these biomarker studies. Here, we describe a protocol for high-density serum and plasma reverse phase protein arrays (RPPAs). We demonstrate how a biobank of 12,392 samples was immobilized and analyzed on a single microarray slide, allowing high-quality profiling of abundant target proteins across all samples in one assay.

Purpose: Our aim was to study the dynamics of the post-surgical canal and nerve volumes and their relationships to objective [electromyoneurography (EMNG)] and subjective (pain) outcomes., Methods: Forty-seven patients with carpal tunnel syndrome (CTS) (median age 52, range 23-75 years) with a prominent narrowing of the median nerve within the canal (observed during carpal tunnel release) were evaluated clinically using EMNG and magnetic resonance imagining (MRI) before and at 90 and 180 days post-surgery., Results: Canal and nerve volumes increased, EMNG findings improved and pain resolved during the follow-up. Increase in tunnel volume was independently associated with increased nerve volume. A greater post-surgical nerve volume was independently associated with a more prominent resolution of pain, but not with the extent of EMNG improvement, whereas EMNG improvement was not associated with pain resolution., Conclusions: Data confirm that MRI can detect even modest changes in the carpal tunnel and median nerve volume and that tunnel release results in tunnel and nerve-volume increases that are paralleled by EMNG and clinical improvements. Taken together, these observations suggest that MRI could be used to objectivise persistent post-surgical difficulties in CTS patients. Level of evidence 3 (follow-up study).

Silver (AgNPs) and maghemite, i.e., superparamagnetic iron oxide nanoparticles (SPIONs) are promising candidates for new medical applications, which implies the need for strict information regarding their physicochemical characteristics and behavior in a biological environment. The currently developed AgNPs and SPIONs encompass a myriad of sizes and surface coatings, which affect NPs properties and may improve their biocompatibility. This study is aimed to evaluate the effects of surface coating on colloidal stability and behavior of AgNPs and SPIONs in modelled biological environments using dynamic and electrophoretic light scattering techniques, as well as transmission electron microscopy to visualize the behavior of the NP. Three dispersion media were investigated: ultrapure water (UW), biological cell culture medium without addition of protein (BM), and BM supplemented with common serum protein (BMP). The obtained results showed that different coating agents on AgNPs and SPIONs produced different stabilities in the same biological media. The combination of negative charge and high adsorption strength of coating agents proved to be important for achieving good stability of metallic NPs in electrolyte-rich fluids. Most importantly, the presence of proteins provided colloidal stabilization to metallic NPs in biological fluids regardless of their chemical composition, surface structure and surface charge. In addition, an assessment of AgNP and SPION behavior in real biological fluids, rat whole blood (WhBl) and blood plasma (BlPl), revealed that the composition of a biological medium is crucial for the colloidal stability and type of metallic NP transformation. Our results highlight the importance of physicochemical characterization and stability evaluation of metallic NPs in a variety of biological systems including as many NP properties as possible.

The proliferation of silver nanoparticle (AgNP) production and use owing to their antimicrobial properties justifies the need to examine the resulting environmental impacts. The discharge of biocidal nanoparticles to water bodies may pose a threat to aquatic species. This study evaluated the effects of citrate-coated AgNPs on the standardized test organism Daphnia magna Straus clone MBP996 by means of biochemical biomarker response. AgNP toxicity was compared against the toxic effect of Ag(+). The toxicity endpoints were calculated based upon measured Ag concentrations in exposure media. For AgNPs, the NOAEC and LOAEC values at 48 h were 5 and 7 μg Ag/L, respectively, while these values were 0.5 and 1 μg Ag/L, respectively, for Ag(+). The EC50 at 48 h was computed to be 12.4 ± 0.6 and 2.6 ± 0.1 μg Ag/L for AgNPs and Ag(+), respectively, with 95 % confidence intervals of 12.1-12.8 and 2.3-2.8 μg Ag/L, respectively. These results indicate significant less toxicity of AgNP compared to free Ag(+) ions. Five biomarkers were evaluated in Daphnia magna neonates after acute exposure to Ag(+) or AgNPs, including glutathione (GSH) level, reactive oxygen species (ROS) content, and catalase (CAT), acetylcholinesterase (AChE), and superoxide dismutase (SOD) activity. AgNPs induced toxicity and oxidative stress responses in D. magna neonates at tenfold higher concentrations than Ag. Biochemical methods revealed a clear increase in AChE activity, decreased ROS level, increased GSH level and CAT activity, but no significant changes in SOD activity. As Ag(+) may dissolve from AgNPs, these two types of Ag could act synergistically and produce a greater toxic response. The observed remarkably high toxicity of AgNPs (in the parts-per-billion range) to crustaceans indicates that these organisms are a vulnerable link in the aquatic food chain with regard to contamination by nanosilver. Graphical Abstract ᅟ.

Aim: To ensure a more reliable classification system for the femur neck fractures by estimating pressure and tensile force on the fracture crevice., Methods: The study compared the kappa coefficient for the inter and intraobserver agreement of the same five observers on the same radiographic sample of the femur neck fracture with a time gap of one year for the AO group and original proposal of estimated dominant pressure and tensile force on the femur neck fracture's crevice., Results: The kappa coefficient for the AO group was 0.49 and for the system for estimating the dominant force on the fracture's crevice it was 0.78. For the observer our proposal is easier to use than the AO system., Conclusion: The estimation system of the dominant force on the fracture's crevice is more reliable and thus more acceptable than the AO group system for femoral neck fractures.

In the treatment of adolescent idiopathic scoliosis, one of the most frequently used technique to determine skeletal maturity is the method described by Risser. The ossification of iliac apophysis progresses from ventral to caudal through the four zones and the fusion of the iliac apophysis to the iliac crest (Risser grade 5) indicated vertebral growth completion, therefore the termination of scoliotic deformity progression. The main disadvantages of Risser method are exposure to radiation and the questionable reliability, so there are efforts to examine iliac apohysis by ultrasound. There is also no resolute recommendation when to discontinue brace treatment of scoliosis. Using ultrasound, in this study, we subdivided Risser grade 4 to grade 4a and 4b, according to the amount of cartilage left unossified, in order to make clear when is safe to end brace treatment. We measured increase in height, during six month period, for 92 healthy children, who were classified by ultrasound in Risser 4a or 4b group. There was significantly larger increase in height for group 4a (p < 0.001). For girls, we also noted time past from menarche as sign of biological maturity. Girls from group Risser 4b got menarche 2.74 years before they were examined while group Risser 4a got menarche only 1.57 years before (p < 0.001). Subdvision of Risser 4 grade by ultrasound is promising method in determining end of brace treatment for scoliosis.

An hip fracture is an significant personal, family and health issue of people older than 65 years. In the first year of the fracture up to 30% of the injured die and about 50% of them never regain their formal degree of independence in fulfilling day-to-day activities. Estimations are that throughout 30 years in the world there will be around 6 million hip fractures per year which is about four times the todays amount. Todays predictions of hip fractures based on the hip geometry have shown us that the hip geometry is an independent variable of the bone mineral density. The hip geometry is more resistant to the effect of various factors than the bone mineral density and the changes throu life are a lot slower. The uniqueness and the sensitivity of the hip geometry in predicting a fracture is high and acceptable in research results of most authors. In this review we present the previous relevant knowledge about the measures and factors which determines the hip geometry and the accepted amount of pictorial methods of hip display. We have compared the methodology and the patients of eleven randomly picked writings on predicting hip fracture based on the hip geometry. We highlight the need of further refinement of the methodology and the more balanced selection of patients for a greater conformity in future writings. The hip geometry has shown it self as an useful diagnostical instrument but there is still more room for its improvement.

Purpose: The purpose of this study was to evaluate the effects of epineurotomy on the post-surgical median nerve volume and clinical outcomes in carpal tunnel syndrome (CTS) patients with a prominent nerve narrowing., Methods: This was a prospective, randomised, double-blind controlled trial. Patients (n = 50) were randomised (1:1) to open-field surgical carpal tunnel release followed by a longitudinal epineurotomy of the nerve (test), or to open-field release without epineurotomy (control)., Results: The nerve volume was slightly larger in the test group 90 days post-surgery (by 10.5 %, p = 0.157) but not 180 days post-surgery. No relevant electropyhsiological or clinical difference between groups and no effect of the nerve volume was observed. The subjective pain reduction was slightly more prominent in the control group at 180 days. Larger post-surgical nerve volume was associated with lower pain, but only in the control group., Conclusions: Even in selected CTS patients, longitudinal epineurotomy confers no benefit regarding the nerve volume or clinical outcomes over a simple carpal tunnel release.

Aim: To determine which of the classification systems for the femoral neck fracture between AO group, AO subgroup, Garden and Pauwels is much more reliable and reproducible to predict a method of treatment, radiological prediction of nonunion and prediction of outcomes., Methods: Five observers classified 77 randomly selected anterior- posterior (AP) and lateral view preoperative radiographs of the femoral neck fractures according to AO group, AO subgroup, Garden and Pauwels classification systems. The procedure was repeated on the same radiographs after three months. First classification is used to calculate interobserver agreement by kappa value between observers, while the first and second classification has served to calculate intraobserver kappa value for each examiner., Results: Overall mean for classification system for interobserver agreement is: AO 0.44, AO subgroup 0.17, Garden 0.41 and Pauwels 0.19. Mean intraobserver agreement for AO group was 0.56, AO subgroup 0.38, Garden 0.49 and Pauwels 0.38 coefficient kappa value., Conclusion: Garden and AO group are useful for the division of femoral neck fractures without displacement and with displacement, but they are not for clinical use. AO subgroup and Pauwels classification are not recommended for further use.

The aim of this retrospective study is to present the incidence of total hip arthroplasty (THA) in Brod-Posavina and Požega-Slavonia counties in 2010 along with an analysis of post-operative complications. The research was carried out at orthopaedic departments in three hospital institutions, "Dr. Josip Benčević "Slavonski Brod General Hospital, the Nova Gradiška General Hospital and the Požega General County Hospital. A total of 27 849 patients was examined in orthopedic outpatient clinics, 1 457 surgical procedures were conducted, of which 239 (16.40%) were THA. There were five (2.09%) dislocations of endoprosthesis, one case (0.42%) of endoprostheses infection, whereas none of periprosthetic fractures and peroneal nerve palsy were presented.

Suprapatellar bursa is located between the quadriceps tendon and femur, and it develops before the birth as a separate synovial compartment proximal to the knee joint. By the fifth month of fetal life there is a suprapatellar septum between the knee joint cavity and suprapatellar bursa which later perforates and involutes in a way that a normal communication between the cavity of bursa and knee is established. A small portion of the embrionic septum can later lag as more or less expressed suprapatellar plica. In case when suprapatellar plica has a small communication with valve mechanism or in case of complete septum, bursa becomes a separate compartment and potential location for the suprapatellar cyst development. Magnetic resonance imaging is recognised as the gold standard in diagnosis of knee cysts because of its ability to show cystic nature of the lesion, its relationship with other anatomic structures, as well as to establish whether other knee pathologies are present. Considering treatment possibilities, majority of cysts around the knee resolve spontaneously and should be treated by aspiration and application of corticosteroids. Suprapatellar cyst is a very rare knee pathology and it can in some occasions be treated using open or arthroscopic surgery.

We will present a rare case of a large symptomatic cyst of the medial meniscus, conjoined with a horizontal tear of a posterior meniscal horn. After clinical, radiological and ultrasound examination, a diagnosis was confirmed with MRI examination of the knee. Literature supports surgical therapy, which includes complete arthroscopic technique, meniscectomy and decompression of the cyst. In our case, it showed that arthroscopic partial meniscectomy and opened cystectomy were the efficient way of treatment.

Carpal tunnel syndrome (CTS) is a somewhat neglected medical and economic problem, and surgery is one of the therapeutic options. We analyze the outcomes of surgical treatment in 114 consecutive patients (154 hands). Before the surgery, physical therapy was implemented (96% cases) and the patients were frequently on a sick leave (42% cases). Immediately before the surgery, the patients suffered intensive pain (median 7 on a 0-10 scale), and had a reduced hand function (median 2 on a 0-10 scale). After the surgery (6-12 months), the pain was reduced (difference -5.0, 95% CL -5.5, -4.5, p<0.001), and the function improved (difference 4.5, 95% CLs 4.0, 5.0, p<0.001). Longer time interval between referral to a primary care physician and referral to an orthopeadic surgeon (>1 year in 48% of the cases) was an independent negative predictor for these outcomes. Total difference in costs for sick leaves and physical therapies between the pre- and postoperative periods was estimated at approximately 269.030,00 to over 375.315,00 euros. The time between the entrance into the healthcare system and recognition of the need for surgical treatment of CTS needs to be reduced in order to get better medical and economic results.