Clinical Outcome of Minimally Manipulated, Mechanically Isolated Autologous Adipose Tissue-Derived Stromal Vascular Fraction (Sahaj Therapy®) in Knee Osteoarthritis-Randomized Controlled Trial.

Introduction: Regenerative therapy has shown promising results in the treatment of osteoarthritis (OA) knee with Kellgren-Lawrence (KL) Grades I-III. We compared the safety, efficacy, functional, and clinical outcomes of intra-articular implantation of autologous adipose tissue-derived stromal vascular fraction (SVF) isolated using direct ultrasonic cavitation (Sahaj therapy-Cell Innovation Patented Technology) and saline injection in knee osteoarthritis. Materials and Methods: The present prospective observational study was conducted over 3 years. We enrolled 120 patients in our study, where four patients got excluded as they did not meet the inclusion criteria. The remaining 116 patients were randomized into two groups, one with autologous adipose tissue-derived SVF and the other group with saline injection. A comparison of mean KOOS and VAS scores at different follow-ups was done using Paired 't' test. A p value of < 0.05 was considered significant. Results: The results show that the SVF group had significantly higher KOOS scores (78.49 ± 6.54 in the SVF group vs 59.19 ± 5.14 in the saline group), respectively (p < 0.001). Similarly, the SVF group had significantly lesser VAS scores (3.17 ± 0.94 in the SVF group vs 3.89 ± 1.04 in the saline group), respectively (p < 0.001). Conclusions: Autologous adipose tissue-derived SVF is a better choice for treating knee osteoarthritis. For individuals with degenerative osteoarthritis, autologous SVF grafting in the same surgical procedure is an innovative and promising treatment modality. Even after 3 years of follow-up, the study participants with OA knee have shown a good clinical and functional outcome.

The Use of Removed Mesquite Brush as a Fiber Replacement in Silage Production.

Mesquite (Prosopis L.) is considered an invasive browse species in most of the American Southwest. Mechanical intervention removes yields an excess of organic debris. Anecdotal evidence in the past has supported using such browse as feed for livestock. Thus, our objectives were to (1) determine the nutritive value and fermentation characteristics of silage produced with mesquite biomass, and (2) evaluate solvent treatment of mesquite biomass prior to ensiling. In Experiment 1, we evaluated mesquite inclusion rate (0, 250, 500, 750, or 1000 g kg−1 DM), length of fermentation (28, 56, or 84 d), and silage inoculant. In Experiment 2, we evaluated the effects of mesquite pre-treatment with acid (H2SO4) or alkali (NaOH) solutions. Concentrations of NDF, ADF, and ADL, as well as IVTD, decreased (p < 0.05) with increasing mesquite inclusion. However, 250 g mesquite kg−1 DM did not differ from grass silage. There was no effect (p > 0.05) of inoculation, though increasing length of incubation did increase (p < 0.05) VFA production and decrease (p < 0.05) silage pH. Solvent treatment did not improve ensiling properties. Results are interpreted to mean that mesquite biomass may be effectively incorporated into silage at levels up to 250 g kg−1.

British Society of Gastroenterology guidelines for the management of iron deficiency anaemia in adults.

Iron deficiency anaemia (IDA) is a major cause of morbidity and burden of disease worldwide. It can generally be diagnosed by blood testing and remedied by iron replacement therapy (IRT) using the oral or intravenous route. The many causes of iron deficiency include poor dietary intake and malabsorption of dietary iron, as well as a number of significant gastrointestinal (GI) pathologies. Because blood is iron-rich it can result from chronic blood loss, and this is a common mechanism underlying the development of IDA-for example, as a consequence of menstrual or GI blood loss.Approximately a third of men and postmenopausal women presenting with IDA have an underlying pathological abnormality, most commonly in the GI tract. Therefore optimal management of IDA requires IRT in combination with appropriate investigation to establish the underlying cause. Unexplained IDA in all at-risk individuals is an accepted indication for fast-track secondary care referral in the UK because GI malignancies can present in this way, often in the absence of specific symptoms. Bidirectional GI endoscopy is the standard diagnostic approach to examination of the upper and lower GI tract, though radiological scanning is an alternative in some situations for assessing the large bowel. In recurrent or refractory IDA, wireless capsule endoscopy plays an important role in assessment of the small bowel.IDA may present in primary care or across a range of specialties in secondary care, and because of this and the insidious nature of the condition it has not always been optimally managed despite the considerable burden of disease- with investigation sometimes being inappropriate, incorrectly timed or incomplete, and the role of IRT for symptom relief neglected. It is therefore important that contemporary guidelines for the management of IDA are available to all clinicians. This document is a revision of previous British Society of Gastroenterology guidelines, updated in the light of subsequent evidence and developments.

Experimental Evaluation of Polyphenylsulfone (PPSF) Powders as Fire-Retardant Materials for Processing by Selective Laser Sintering.

Additive manufacturing has progressed rapidly, and the unique attributes of the layer-wise material consolidation are attracting ever increasing application potentials in critical sectors such as medical and aerospace industries. A lack of materials options has been the main bottleneck for the much wider uptake of these promising new technologies. Inventing new material alternatives has been central to most of the research attention in additive manufacturing in recent times. The current research is focused on evaluating the polyphenylsulfone polymer powders for the first time as fire-resistant candidate materials for processing by selective laser sintering, the most promising additive processing method for polymeric material systems. Experimental evaluations were undertaken based on a selective laser sintering test bed. Single layer and multi-layer samples were produced for microstructural and mechanical characterisations. The microstructural evaluations and the mechanical property results indicate sufficient intra- and inter-layer consolidation together with reasonable tensile property responses. The lower viscosity and thermal conductivity characteristics rendered lower tensile strengths, which will require some further attention in the future, for better consolidation and mechanical properties.

Imaging Mutant Huntingtin Aggregates: Development of a Potential PET Ligand.

Mutant huntingtin (mHTT) protein carrying the elongated N-terminal polyglutamine (polyQ) tract misfolds and forms protein aggregates characteristic of Huntington's disease (HD) pathology. A high-affinity ligand specific for mHTT aggregates could serve as a positron emission tomography (PET) imaging biomarker for HD therapeutic development and disease progression. To identify such compounds with binding affinity for polyQ aggregates, we embarked on systematic structural activity studies; lead optimization of aggregate-binding affinity, unbound fractions in brain, permeability, and low efflux culminated in the discovery of compound 1, which exhibited target engagement in autoradiography (ARG) studies in brain slices from HD mouse models and postmortem human HD samples. PET imaging studies with 11C-labeled 1 in both HD mice and WT nonhuman primates (NHPs) demonstrated that the right-hand-side labeled ligand [11C]-1R (CHDI-180R) is a suitable PET tracer for imaging of mHTT aggregates. [11C]-1R is now being advanced to human trials as a first-in-class HD PET radiotracer.

Differential Gene Expression of Lymphocytes Stimulated with Rhinovirus A and C in Children with Asthma.

Rationale: Individuals with asthma have heightened antibody responses to rhinoviruses (RVs), although those specific for RV-C are lower than responses specific for RV-A, suggesting poor immunity to this species.Objectives: To ascertain and compare T-cell memory responses induced by RV-A and RV-C in children with and without asthma.Methods: Peripheral blood mononuclear cells from 17 children with asthma and 19 control subjects without asthma were stimulated in vitro with peptide formulations to induce representative species-specific responses to RV-A and RV-C. Molecular profiling (RNA sequencing) was used to identify enriched pathways and upstream regulators.Measurements and Main Results: Responses to RV-A showed higher expression of IFNG and STAT1 compared with RV-C, and significant expression of CXCL9, 10, and 11 was not found for RV-C. There was no reciprocal increase of T-helper cell type 2 (Th2) cytokine genes or the Th2 chemokine genes CCL11, CCL17, and CCL22. RV-C induced higher expression of CCL24 (eotaxin-2) than RV-A in the responses of children with and without asthma. Upstream regulator analysis showed both RV-A and, although to a lesser extent, RV-C induced predominant Th1 and inflammatory cytokine expression. The responses of children with asthma compared with those without asthma were lower for both RV-A and RV-C while retaining the pattern of gene expression and upstream regulators characteristic of each species. All groups showed activation of the IL-17A pathway.Conclusions: RV-C induced memory cells with a lower IFN-γ-type response than RV-A without T-helper cell type 2 (Th2) upregulation. Children with asthma had lower recall responses than those without asthma while largely retaining the same gene activation profile for each species. RV-A and RV-C, therefore, induce qualitatively different T-cell responses.

Effects of Ser47-Point Mutation on Conformation Structure and Allergenicity of the Allergen of Der p 2, a Major House Dust Mite Allergen.

PURPOSE: Hypoallergenic recombinant Der p 2 has been produced by various genetic manipulations, but mutation of a naturally polymorphic amino acid residue known to affect IgE binding has not been studied. This study aimed to determine the effect of a point mutation (S47W) of residue 47 of Der p 2 on its structure and immunoglobulin (Ig) E binding. Its ability to induce pro-inflammatory responses and to induce blocking IgG antibody was also determined. METHODS: S47 of recombinant Der p 2.0110, one of the predominant variants in Bangkok, was mutated to W (S47W). S47W secreted from Pichia pastoris was examined for secondary structure and for the formation of a hydrophobic cavity by 8-Anilino-1-naphthalenesulfonic acid (ANS) staining. Monoclonal and human IgE-antibody binding was determined by enzyme-linked immunosorbent assay. Allergen-induced degranulation by human epsilon receptor expressed-rat basophil was determined. Stimulation of the pro-inflammatory cytokine interleukin (IL)-8 release from human bronchial epithelial (BEAS2B) cells and inhibition of IgE binding to the wild type allergen by S47W-induced IgG were determined. RESULTS: S47W reduced secondary structure and failed to bind the hydrophobic ANS ligand as well as a monoclonal antibody known to be dependent on the nature of the side chain of residue 114 in an adjacent loop. It could also not stimulate IL-8 release from BEAS2B cells. IgE from house dust mite (HDM)-allergic Thais bound S47W with 100-fold weaker avidity, whereas IgE of HDM-allergic Australians did not. S47W still induced basophil degranulation, although requiring higher concentrations for some subjects. Anti-S47W antiserum-immunized mice blocked the binding of human IgE to wild type Der p 2. CONCLUSIONS: The mutant S47W had altered structure and reduced ability to stimulate pro-inflammatory responses and to bind IgE, but retained its ability to induce blocking antibodies. It thus represents a hypoallergen produced by a single mutation of a non-solvent-accessible amino acid.

Germline pharmacogenomics of DPYD*9A (c.85T>C) variant in patients with gastrointestinal malignancies treated with fluoropyrimidines.

BACKGROUND: The correlation between DPYD*9A (c.85T>C) genotype and dihydropyrimidine dehydrogenase (DPD) deficiency clinical phenotype is controversial. Reference laboratories either did not perform DPYD*9A genotyping or have stopped DPYD*9A genotyping and limited genotyping to high-risk variants (DPYD*2A, DPYD*13 and DPYD*9B) only. This study explored DPYD*9A genotype and clinical phenotype correlation in patients with gastrointestinal (GI) malignancies treated with fluoropyrimidines. METHODS: Between 2011 and 2017, 67 patients with GI malignancies were genotyped for DPYD variants. Fluoropyrimidines-associated toxicity was graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events (version 3.0). Fisher's exact test was used for statistical analysis. RESULTS: DPYD variants were identified in 17 out of 67 (25%) patients. One patient was homozygous for DPYD*9A variant and one patient was double heterozygous for DPYD*9A and DPYD*9B variants. In patients with identified DPYD variants, 13/17 (76%) patients had DPYD*9A variant, 3/17 (18%) patients had DPYD*2A variant and 2/17 (12%) patient had DPYD*9B variant. Only patients genotyped prior to 2015 were genotyped for DPYD*9A variant (N=28). Of those, 13/28 patients (46%) had DPYD*9A variant. Grade 3-4 diarrhea was associated with DPYD*9A variant in patients treated with full dose fluoropyrimidines (P=0.0055). CONCLUSIONS: In our cohort, DPYD*9A variant was the most common diagnosed variant. The correlation between DPYD*9A genotype and DPD deficiency in clinical phenotype was noticeable in patients who received full dose fluoropyrimidines as they all experienced grade 3-4 toxicities (diarrhea).

An Evaluation of On-Call Otolaryngology Consultations: Assessing an Increasing Workload.

OBJECTIVE: This study compares the volume of on-call otolaryngology consultations in a tertiary care center over a 5-year period. The objective of this study was to identify changes in the volume of consultations in an inpatient setting. METHODS: A cross-sectional retrospective study was performed to determine the volume of consultations. The years 2010 and 2015 were the timepoints for the cross-sectional analysis. A review of electronic medical records was performed to identify all patients associated with the otolaryngology service from the emergency department, inpatient wards, and intensive care units. The primary outcome was the number of otolaryngology consultations per year. RESULTS: The number of on-call consultations in 2010 was 992. In 2015, the number of on-call consultations was 2174. This represents a 120% increase in the number of consultations over a 5-year period ( P < .001). CONCLUSION: There has been a significant increase in the volume of on-call otolaryngology consultations at our tertiary care center. This increase has the potential to adversely affect patient care. A better understanding of the cause of this increase may allow policymakers and health care practitioners to improve patient access, physician workloads, and resource allocation.

WHO/IUIS Allergen Nomenclature: Providing a common language.

A systematic nomenclature for allergens originated in the early 1980s, when few protein allergens had been described. A group of scientists led by Dr. David G. Marsh developed a nomenclature based on the Linnaean taxonomy, and further established the World Health Organization/International Union of Immunological Societies (WHO/IUIS) Allergen Nomenclature Sub-Committee in 1986. Its stated aim was to standardize the names given to the antigens (allergens) that caused IgE-mediated allergies in humans. The Sub-Committee first published a revised list of allergen names in 1986, which continued to grow with rare publications until 1994. Between 1994 and 2007 the database was a text table online, then converted to a more readily updated website. The allergen list became the Allergen Nomenclature database (www.allergen.org), which currently includes approximately 880 proteins from a wide variety of sources. The Sub-Committee includes experts on clinical and molecular allergology. They review submissions of allergen candidates, using evidence-based criteria developed by the Sub-Committee. The review process assesses the biochemical analysis and the proof of allergenicity submitted, and aims to assign allergen names prior to publication. The Sub-Committee maintains and revises the database, and addresses continuous challenges as new "omics" technologies provide increasing data about potential new allergens. Most journals publishing information on new allergens require an official allergen name, which involves submission of confidential data to the WHO/IUIS Allergen Nomenclature Sub-Committee, sufficient to demonstrate binding of IgE from allergic subjects to the purified protein.

IgE and T-cell responses to house dust mite allergen components.

Using the terminology for Dermatophagoides pteronyssinus, IgE responses to house dust mites have been shown to be mostly directed to the serodominant Der p 1, 2 and 23 allergen components with mid -tier responses to Der p 4, 5, 7 and 21 that are made by 30-50% of subjects with titers proportional to those of the serodominant specificities. This pattern can be seen to evolve in childhood and although responses to minor allergens appear to contribute little to the total IgE they are at least markers for a greater propensity to develop disease. While Der p 23 is a component that induces prevalent IgE responses, sometimes in the absence of responses to Der p 1and 2, not all studies have found high titers so further investigation is needed. From limited knowledge adult onset IgE responses might have a different pattern that is not so centered on Der p 1 and 2. Responses that induce under 3.5 IU/ml of IgE antibody are not usually associated with disease and should be examined for cross reactivity expected from IgE responses to other allergens and antigens of infectious agents. Scabies that has 40% endemicity in some regions and is spread by immigration can give rise to high-titer binding that can be recognized by component resolved diagnosis. Recent studies with synthetic peptides representing allergens and non-allergenic HDM proteins now offer new research avenues on HDM induced immune responses, including the ability to use peptides representing the serodominant allergens as defined reagents for long overdue reproducible T-cell investigations.

T-cell responses against rhinovirus species A and C in asthmatic and healthy children.

BACKGROUND: Infections by rhinovirus (RV) species A and C are the most common causes of exacerbations of asthma and a major cause of exacerbations of other acute and chronic respiratory diseases. Infections by both species are prevalent in pre-school and school-aged children and, particularly for RV-C, can cause severe symptoms and a need for hospitalization. While associations between RV infection and asthma are well established, the adaptive immune-mechanisms by which RV infections influence asthma exacerbations are yet to be defined. OBJECTIVE: The aim of this study was to characterize and compare T-cell responses between RV-A and RV-C and to test the hypothesis that T-cell responses would differ between asthmatic children and healthy controls. METHODS: A multi-parameter flow cytometry assay was used to characterize the in vitro recall T-cell response against RV-A and RV-C in PBMCs from children with acute asthma (n = 22) and controls (n = 26). The responses were induced by pools of peptides containing species-specific VP1 epitopes of RV-A and RV-C. RESULTS: Regardless of children's clinical status, all children that responded to the in vitro stimulation (>90%) had a similar magnitude of CD4+ T-cell responses to RV-A and RV-C. However, asthmatic children had a significantly lower number of circulating regulatory T cells (Tregs), and healthy controls had significantly more Tregs induced by RV-A than RV-C. CONCLUSIONS AND CLINICAL RELEVANCE: The comparable recall memory T-cell responses in asthmatic and control children to both RV-A and RV-C show that differences in the antibody and inflammatory responses previously described are likely to be due to regulation, with a demonstrated candidate being reduced regulatory T-cells. The reduced Treg numbers demonstrated here could explain the asthmatic's inability to appropriately control immunopathological responses to RV infections.

Prominent IgE-binding and cytokine-inducing capacities of a newly cloned N-terminal region of Der f 14, an apolipophorin-like house dust mite allergen.

We previously characterized a 177-kDa allergen, M-177, from Dermatophagoides farinae. Thereafter, a counterpart to M-177 for Euroglyphus maynei was cloned as Eur m 14, and its sequence revealed that two environmental allergens, Mag 1 and Mag 3, are digested fragments of M-177. The aims of this study were to clone the cDNA of Der f 14 corresponding to M-177 and to elucidate the allergenic capacities of the N-terminal fragment of Der f 14 (Der f 14-N). Recombinant allergens were produced as trigger-factor-fused proteins in Escherichia coli. Der f 14-N showed the highest IgE-binding frequency among Der f 14-derived fragments in patients allergic to house dust mite by enzyme-linked immunosorbent assay. Der f 14-N showed the highest capacity to induce cell proliferation in murine lymphocyte and human peripheral mononuclear cells among Der f 14-derived fragments. Der f 14-N induced IL-13, IFN-γ and IL-17 production more than Der f 1 and Der f 2 in mouse, and induced IL-5 and IFN-γ production at levels comparable to those of Der f 1 and Der f 2 in some patients. The high prevalence of IgE binding to the Der f 14-N indicates that it could be an important mite allergen.

International consensus (ICON) on: clinical consequences of mite hypersensitivity, a global problem.

Since mite allergens are the most relevant inducers of allergic diseases worldwide, resulting in significant morbidity and increased burden on health services, the International Collaboration in Asthma, Allergy and Immunology (iCAALL), formed by the American Academy of Allergy, Asthma and Immunology (AAAAI), the American College of Allergy, Asthma and Immunology (ACAAI), the European Academy of Allergy and Clinical Immunology (EAACI), and the World Allergy Organization (WAO), has proposed to issue an International Consensus (ICON) on the clinical consequences of mite hypersensitivity. The objectives of this document are to highlight aspects of mite biology that are clinically relevant, to update the current knowledge on mite allergens, routes of sensitization, the genetics of IgE responses to mites, the epidemiologic aspects of mite hypersensitivity, the clinical pictures induced by mites, the diagnosis, specific immunotherapeutic approaches, and prevention.

Propiedades biológicas de los alérgenos inhalatorios y su relación con la respuesta inmunitaria / Biological properties of inhalant aeroallergens and their relation to immune responses

IgE-binding studies show that many of the common causes of inhalant allergy such as grass, olive, ragweed and birch pollen, house dust mites and some fungi have one or a few principal allergens that can account for most of the allergic response. The IgE binding to allergens from other sources can be more evenly spread amongst different proteins or, as indicated in cat allergy, varies with clinical presentation. The biological properties of nearly all of the principal allergens can now be predicted from the knowledge of their structures and they point to likely interactions with the innate immune system, as well as possible interactions with hormonal regulators of immunity. As found for pectate lyases and the Ole e1-like proteins, biologically similar proteins can be principal allergens for many species while the Dermatophagoides spp. and Blomia tropicalis allergens show that allergens with the same biological properties reveal interspecies variation in allergen hierarchy. These properties show that the interactions of allergens with innate immunity and immuno-regulators will be different for different allergens, and this concurs with the evidence that immune responses to allergens from the same source are regulated independently, as are responses to co-presented allergenic and non-allergenic proteins

Los estudios de unión con la IgE demostraron que muchas de las causas comunes de alergia inhalatoria, como a las gramíneas, el olivo, la ambrosía, el polen de abedul, los ácaros del polvo doméstico y algunos hongos, tienen uno o unos pocos de los alérgenos principales que pueden representar la mayoría de las respuestas alérgicas. La IgE que se une a los alérgenos de otras fuentes puede diseminarse entre diferentes proteínas o, como indica la alergia al gato, varía con la presentación clínica. Las propiedades biológicas de casi todos los alérgenos principales pueden actualmente predecirse a partir del conocimiento de sus estructuras e indican las interacciones probables con el sistema inmunitario innato, así como las interacciones posibles con los reguladores hormonales de la inmunidad. Como se encontró para las pectato liasas y las proteínas similares a Ole e1, las proteínas biológicamente similares pueden ser los alérgenos principales para muchas especies, mientras que los alérgenos Dermatophagoides spp. y Blomia tropicalis muestran que los alérgenos con las mismas propiedades biológicas tienen variación entre las especies en la jerarquía alergénica. Estas propiedades demuestran que las interacciones de los alérgenos con la inmunidad innata y los inmunorreguladores serían diferentes para los distintos alérgenos, y esto coincide con las pruebas que indican que las respuestas inmunes a los alérgenos de la misma fuente sufren una regulación por aumento (upregulation), independientemente de si son respuestas a las proteínas co-presentadas alergénicas y no alergénicas

Immunodominant T-Cell Epitopes in the VP1 Capsid Protein of Rhinovirus Species A and C.

Rhinovirus (RV) species A and C are the most frequent cause of respiratory viral illness worldwide, and RV-C has been linked to more severe exacerbations of asthma in young children. Little is known about the immune responses to the different RV species, although studies comparing IgG1 antibody titers found impaired antibody responses to RV-C. Therefore, the aim of this study was to assess whether T-cell immunity to RV-C is similarly impaired. We measured T-cell proliferation to overlapping synthetic peptides covering the entire VP1 capsid protein of an RV-A and RV-C genotype for 20 healthy adult donors. Human leukocyte antigen (HLA) was typed in all the donors in order to investigate possible associations between the HLA type and RV peptide recognition. Total and specific IgG1 antibody titers to the VP1 proteins of both RV-A and RV-C were also measured to examine associations between the antibody and T-cell responses. We identified T-cell epitopes that are specific to and representative of each RV-A and RV-C species. These epitopes stimulated CD4+-specific T-cell proliferation, with similar magnitudes of response for both RV species. All the donors, independent of their HLA-DR or -DQ type, were able to recognize the immunodominant RV-A and -C regions of VP1. Furthermore, the presence or absence of specific antibody titers was not related to changes in T-cell recognition. Our results indicate a dissociation between the antibody and T-cell responses to rhinoviruses. The species-representative T-cell epitopes identified in this study are valuable tools for future studies investigating T-cell responses to the different RV species. IMPORTANCE: Rhinoviruses (RVs) are mostly associated with the common cold and asthma exacerbations, although their contributions to most upper and lower respiratory tract diseases have increasingly been reported. Species C (RV-C) has been associated with more frequent and severe asthma exacerbations in young children and, along with RV-A, is the most clinically relevant species. Little is known about how our immune system responds to rhinoviruses, and there are limited tools to study specific adaptive immunity against each rhinovirus species. In this study, we identified immunodominant T-cell epitopes of the VP1 proteins of RV-A and RV-C, which are representative of each species. The study found that T-cell responses to RV-A and RV-C were of similar magnitudes, in contrast with previous findings showing RV-C-specific antibody responses were low. These findings will provide the basis for future studies on the immune response to rhinoviruses and can help elucidate the mechanisms of severity of rhinovirus-induced infections.