R-mandelonitrile-lyase, homolog to Pru d 10, is a major peach allergen in peach allergic Spanish population.

BACKGROUND AND OBJECTIVE: Peach allergy is a prevalent cause of food allergy. Despite the repertoire of allergens available for molecular diagnosis, there are still patients with undetectable IgE levels to peach allergens but presenting symptoms after its ingestion. The objective of this study was to investigate the allergenic profile in a patient population with symptoms produced by peach. METHODS: An exploratory retrospective study was performed with patients presenting symptoms after the ingestion of peach. Forty-two patients were included in the study. The allergenic profile of individual patients was investigated by immunoblot. A serum pool was prepared with the sera that recognized a 70 kDa band. This pool was used to detect this protein in peach peel and pulp and to identify the 70 kDa protein in 2D immunoblot. Spots recognized in the 2D immunoblot were sequenced by LC-MS/MS. Inhibition studies were performed between peach peel and almond. RESULTS: Twenty-two patients (52.4%) recognized the 70 kDa protein in immunoblot. This protein was recognized in peel and pulp. Two different spots were observed in 2D-PAGE, both were identified as (R)-mandelonitrile lyases (RML) with high amino acid similarity with Pru du 10. Peach RML were partially inhibited with an almond extract. No association was found between any reported symptom and sensitization to RML. RML-sensitized patients were older and reported pollen associated respiratory symptoms more frequently than negative patients. CONCLUSION: A new peach allergen, a RML, homologous of Pru du 10, recognized by 52% of the population has been identified.

Sensitization to Vitis vinifera Pollen in a Wine Production Area: Identification of the Allergens Involved.

BACKGROUND: Vine cultivation is widely distributed in La Rioja, Spain (37% of all crops) and is associated with exposure of the general population to vine pollen. The aims of this study were to investigate the prevalence of sensitization to Vitis vinifera pollen in persons with respiratory allergy in the general population and to identify the allergens involved. MATERIALS AND METHODS: The study population comprised patients who came to the hospital between September 2019 and January 2020 with suspected respiratory allergy. All patients underwent skin prick testing with a panel of standardized aeroallergens, profilin, lipid transfer protein (LTP), and V vinifera pollen extract and prick-prick testing with fresh grapes. The in vitro study included specific IgE by ImmunoCap and ELISA, allergenic profile by immunoblot with individual sera from patients positive to V vinifera pollen extract, and 2D immunoblot with a pool of sera. The spots recognized by IgE were identified using mass spectrometry. RESULTS: A total of 151 patients were included. Of these, 124 were positive to some of the allergens tested. Thirty-four (27.4%) were positive to vine pollen in the skin prick tests. The serology study revealed positive results in 20 patients. Five vine pollen allergens were identified, and profilin was the most prevalent (30%). The other 4 allergens could be considered specific to this pollen. CONCLUSIONS: Sensitization to vine pollen was frequent in the general population in a vine growing area. The clinical relevance of this finding is unknown owing to sensitization to other pollens in the vine pollen-positive patients. Five new vine pollen allergens were identified.

The Connection Between Stress and Immune Status in Pigs: A First Salivary Analytical Panel for Disease Differentiation.

This paper analyzes the association between stress and immune response activations in different diseases, based on the salivary analytics. Moreover, a first attempt to discriminate between diseases was performed by principal component analysis. The salivary analytics consisted of the measurement of psychosocial stress (cortisol and salivary alpha-amylase) indicators, innate (acute phase proteins: C-reactive protein and haptoglobin), and adaptive immune (adenosine deaminase, Cu and Zn) markers and oxidative stress parameters (antioxidant capacity and oxidative status). A total of 107 commercial growing pigs in the field were divided into six groups according to the signs of disease after proper veterinary clinical examination, especially, healthy pigs, pigs with rectal prolapse, tail-biting lesions, diarrhea, lameness, or dyspnea. Associations between stress and immune markers were observed with different intensities. High associations (r = 0.61) were observed between oxidative stress markers and adaptive immune markers. On the other hand, moderate associations (r = 0.31-0.48) between psychosocial stress markers with both innate and adaptive immune markers were observed. All pathological conditions showed statistically significant differences in at least 4 out of the 11 salivary markers studied, with no individual marker dysregulated in all the diseases. Moreover, each disease condition showed differences in the degree of activation of the analyzed systems which could be used to create different salivary profiles. A total of two dimensions were selected through the principal component analysis to explain the 48.3% of the variance of our data. Lameness and rectal prolapse were the two pathological conditions most distant from the healthy condition followed by dyspnea. Tail-biting lesions and diarrhea were also far from the other diseases but near to healthy animals. There is still room for improvements, but these preliminary results displayed a great potential for disease detection and characterization using salivary biomarkers profiling in the near future.

Lesiones de Lisfranc: repercusión clínico-funcional / Lisfranc lesions: clinical-functional repercussion

Resumen: Objetivo: Valoración clínica-funcional de pacientes con lesión de Lisfranc sin causa laboral ni deportiva tratados en nuestro centro. Material y métodos: Se analizaron 13 pacientes con lesión de Lisfranc sin causa laboral ni deportiva, divididos en dos grupos, lesiones de alta energía y de baja energía. Valoración del retorno a la actividad laboral y deportiva previa a la lesión y el uso de calzado y plantillas posteriormente. Se recogen las puntuaciones del test AOFAS y EVA postoperatorio así como la aparición de complicaciones posteriores a la recuperación de la lesión. Resultados: El grupo de alta energía lo formaron siete pacientes que fueron intervenidos en una media de 7.3 días; cuatro presentaron secuelas postquirúrgicas. De estos pacientes, un paciente no pudo reincorporarse a su actividad laboral previa y dos pacientes no pudieron realizar el deporte que practicaban previamente a la lesión. El AOFAS fue de 79.4 y el EVA de 2.8. En el grupo de baja energía encontramos seis pacientes intervenidos en 5.6 días de media. Todos ellos pudieron reincorporarse a su actividad laboral y deportiva previa. El AOFAs fue de 84.8 y el EVA de 1.6. En ninguna de las variables estudiadas se observaron diferencias estadísticamente significativas. Conclusión: Los pacientes con lesión de Lisfranc fuera del ámbito laboral y deportivo presentan buenos resultados clínicos y funcionales.

Abstract: Objective: Clinical and functional evaluation of patients with Lisfranc lesion without work or sport cause and treated in our center. Material and methods: Thirteen patients with Lisfranc lesion without occupational or sports causes were divided into two groups, high-energy and low-energy injuries. Were analyzed the return to work and sports activity prior to the injury and the use of footwear and insoles after the injury. Also were assessed AOFAS and EVA postoperative score and posterior complications after lesion recovery. Results: High-energy group included 7 patients who underwent surgery on an average of 7.3 days. Of the total of patients, 4 presented post-surgical sequelae. Of these patients, one patient was unable to return to their previous work activity and two patients were unable to practice the usual sports activity. The AOFAS was 79.4 and EVA 2.8. In the low energy group, we found six patients operated on 5.6 days on average. All of them were able to return to their previous work and sports activity. The AOFAS were 84.8 and the EVA 1.6. Statistically significant differences were not observed in any of the variables studied. Conclusion: Patients with Lisfranc injury out of laboral or sports background present good clinical and functional results.

Immunoinformatic epitope prediction to select monoclonal antibodies for Phl p 1 quantification.

BACKGROUND: Allergen quantification has become a relevant parameter for allergen extract characterization and to guarantee the consistency of the manufacturing process at allergen immunotherapy. The aim of this study was to develop and validate a method to quantify the major allergen Phl p 1 based on a prediction of the antigenic regions by immunoinformatic strategies. METHODS: Phl p 1 was purified from a Phleum pratense native extract by chromatographic methods. Immunoinformatic tools were used to predict B-cell epitopes. In silico predictions were verified by mapping linear epitopes with a peptide library and used to select the appropriate regions for producing the mAbs to develop an ELISA method, which was validated. Phl p 1 was quantified in 24 batches of P. pratense extracts. RESULTS: Phl p 1 was purified with 95 % purity and completely functional. Eight B-cell epitopes in each of the two Phl p 1 isoforms were predicted. Two of the predicted B-cell epitopes overlapped with the experimentally determined peptides recognized by two mAbs selected for development of the kit. The quantification method demonstrated to be specific to Phl p 1, linear, accurate and precise in the range from 7.7 to 123.3 µg/mg. Mean Phl p 1 content was 28.95 µg of allergen/mg of lyophilized native extract and 44.23 µg of allergen/mg of lyophilized depigmented extract. CONCLUSIONS: An ELISA method for measuring Phl p 1 in P. pratense extracts was developed and validated by producing the appropriate mAbs against epitopes selected by immunoinformatic tools.

[Lisfranc lesions: clinical-functional repercussion]. / Lesiones de Lisfranc: repercusión clínico-funcional.

OBJECTIVE: Clinical and functional evaluation of patients with Lisfranc lesion without work or sport cause and treated in our center. MATERIAL AND METHODS: Thirteen patients with Lisfranc lesion without occupational or sports causes were divided into two groups, high-energy and low-energy injuries. Were analyzed the return to work and sports activity prior to the injury and the use of footwear and insoles after the injury. Also were assessed AOFAS and EVA postoperative score and posterior complications after lesion recovery. RESULTS: High-energy group included 7 patients who underwent surgery on an average of 7.3 days. Of the total of patients, 4 presented post-surgical sequelae. Of these patients, one patient was unable to return to their previous work activity and two patients were unable to practice the usual sports activity. The AOFAS was 79.4 and EVA 2.8. In the low energy group, we found six patients operated on 5.6 days on average. All of them were able to return to their previous work and sports activity. The AOFAS were 84.8 and the EVA 1.6. Statistically significant differences were not observed in any of the variables studied. CONCLUSION: Patients with Lisfranc injury out of laboral or sports background present good clinical and functional results.

OBJETIVO: Valoración clínica-funcional de pacientes con lesión de Lisfranc sin causa laboral ni deportiva tratados en nuestro centro. MATERIAL Y MÉTODOS: Se analizaron 13 pacientes con lesión de Lisfranc sin causa laboral ni deportiva, divididos en dos grupos, lesiones de alta energía y de baja energía. Valoración del retorno a la actividad laboral y deportiva previa a la lesión y el uso de calzado y plantillas posteriormente. Se recogen las puntuaciones del test AOFAS y EVA postoperatorio así como la aparición de complicaciones posteriores a la recuperación de la lesión. RESULTADOS: El grupo de alta energía lo formaron siete pacientes que fueron intervenidos en una media de 7.3 días; cuatro presentaron secuelas postquirúrgicas. De estos pacientes, un paciente no pudo reincorporarse a su actividad laboral previa y dos pacientes no pudieron realizar el deporte que practicaban previamente a la lesión. El AOFAS fue de 79.4 y el EVA de 2.8. En el grupo de baja energía encontramos seis pacientes intervenidos en 5.6 días de media. Todos ellos pudieron reincorporarse a su actividad laboral y deportiva previa. El AOFAs fue de 84.8 y el EVA de 1.6. En ninguna de las variables estudiadas se observaron diferencias estadísticamente significativas. CONCLUSIÓN: Los pacientes con lesión de Lisfranc fuera del ámbito laboral y deportivo presentan buenos resultados clínicos y funcionales.

Profilin is a marker of severity in allergic respiratory diseases.

BACKGROUND: The capacity of profilin to induce allergic symptoms in patients with respiratory allergy has been questioned. In this sense, the aim of this study was to investigate the correlation between profilin exposure and induction of symptoms in a prospective case-control study. METHODS: The concentration of profilin as well as pollen levels in the air was measured. A diary score of symptoms was collected from allergic patients. Seventy-nine individuals were included in the study; fifty cases and 28 controls were positive or negative to profilin, respectively. Conjunctival and bronchial provocation tests were performed with purified profilin (Pho d 2) in a subgroup of cases and controls. RESULTS: Profilin was detected in the environment on 133 days (maximum peak of 0.56 ng/m3 ). A positive correlation between profilin and pollen count of Olea and Poaceae was observed (ρ = 0.24; P < .001). Intensity of total, nasal and ocular symptoms was statistically higher in cases than in controls (P < .001). The risk of suffering symptoms, measured by the percentage of patients who presented any of the symptoms each day, was also higher in cases than in controls. The provocation test was positive in 95% of bronchial and 90% of conjunctival challenges in cases, and negative in all controls. CONCLUSIONS: Profilin was detected in the environment and had the ability to induce a specific allergen response. Patients sensitized to this panallergen showed more symptoms and were more likely to have symptoms. Therefore, sensitization to profilin seems to be a marker of severity in patients with rhinoconjunctivitis and asthma mediated by pollen.

How to interpret elevated plasmatic level of high-sensitive troponin T in newborns and infants?

Research and clinical implications on novel cardiac biomarkers has intensified significantly in the past few years. The high-sensitive troponin T (hscTnT) assay plays a dominant role in diagnostic algorithm regarding myocardial injury in adults. Despite generally accepted use of hscTnT there are no data about physiological concentrations and cut-off limits in neonates and infants to date. The aim of this study is to assess hscTnT levels in healthy newborns and infants. Consecutively 454 healthy full termed newborns and 40 healthy infants were enrolled in the study. Samples of cord or venous blood were drawn and tested for hscTnT concentrations with high-sensitive TnT assay (Roche Cobas e602 immunochemical analyzer). The 97.5 percentile of hscTnT concentration was assessed and correlation analysis was performed in neonates. Two hundred and thirteen samples (47 %) were excluded due to blood hemolysis of various degrees in neonates. Finally, the group of 241 healthy newborns was statistically analyzed. The median concentration of hscTnT was 38.2 ng/ml, 97.5 percentile reached 83.0 ng/l (confidential interval 74.1 to 106.9 ng/l). HscTnT concentrations were statistically decreased in hemolytic samples when compared to non-hemolytic samples (34.3 ng/l [26.7 to 42.0 ng/l] and 37.1 ng/l [30.5 to 47.9 ng/l], respectively, p=0.003). Elevated plasma concentrations of hscTnT decreased to adult level within six months. This study has confirmed the higher reference levels of hscTnT in neonates and young infants when compared with adult population. Many extracardiac factors as hemolysis and age may affect the hscTnT level. Based on presented results, a careful clinical interpretation of hscTnT is recommended.

Pre-operative haemoglobin levels and iron status in a large multicentre cohort of patients undergoing major elective surgery.

Pre-operative anaemia in patients undergoing major surgical procedures has been linked to poor outcomes. Therefore, early detection and treatment of pre-operative anaemia is recommended. However, to effectively implement a pre-operative anaemia management protocol, an estimation of its prevalence and main causes is needed. We analysed data from 3342 patients (44.5% female) scheduled for either: elective orthopaedic surgery (n = 1286); cardiac surgery (n = 691); colorectal cancer resection (n = 735); radical prostatectomy (n = 362); gynaecological surgery (n = 203) or resection of liver metastases (n = 122). For both sexes, anaemia was defined by a haemoglobin level < 130 g.l-1 ; absolute iron deficiency by ferritin < 30 ng.ml-1 (< 100 ng.ml-1 , if transferrin saturation < 20% or C-reactive protein > 5 mg.l-1 ); iron sequestration by transferrin saturation < 20% and ferritin > 100 ng.ml-1 ; and low iron stores by transferrin saturation > 20% and ferritin 30-100 ng.ml-1 . The overall prevalence of anaemia was 36%, with differences according to the type of surgery. Laboratory parameters allowing classification of iron status were available for 2884 patients. Among those with anaemia (n = 986), 677 (69%) were women, 608 (62%) presented with absolute iron deficiency, 101 (10%) with iron sequestration; and 150 (5%) with low iron stores. Iron status alterations were similar in women with haemoglobin < 130 g.l-1 or < 120 g.l-1 . For those who were not anaemic (n = 1898), corresponding figures were 656 (35%), 621 (33%), 165 (9%) and 518 (27%), respectively. Anaemia was present in one-third of patients undergoing major elective procedures. Over two-thirds of anaemic patients presented with absolute iron deficiency or iron sequestration. Over half of non-anaemic patients presented with absolute iron deficiency or low iron stores. We consider these data useful for planning pre-operative management of patients scheduled for major elective surgery.

Purification and immunochemical characterization of Pla l 2, the profilin from Plantago lanceolata.

Profilins are small actin-binding proteins found in eukaryotes and involved in cell development, cytokinesis, membrane trafficking, and cell motility. From an allergenic point of view, profilins are panallergens usually involved in allergic polysensitization, although they are generally recognized as minor allergens. The objectives of this study were to identify and characterize the profilin from Plantago lanceolata pollen and to investigate the cross-reactivity between profilins from different pollen allergenic sources. Profilins from P. lancelolata (Pla l 2) and palm tree pollen (Pho d 2) were purified by affinity chromatography, deeply characterized and identified by mass spectrometry. Pla l 2 allergenicity was confirmed by immunoblot with serum samples from a patient population sensitized to profilin. Immunoblot inhibition was performed to study IgG reactivity between different pollen profilins. IgE cross-reactivity was demonstrated by ImmunoCAP inhibition. Pla l 2 is the second P. lanceolata allergen included in the IUIS Allergen Nomenclature database. Four peptides from purified Pla l 2 were identified with percentages of homology with other pollen profilins between 73 and 86%. Eighty-six percent (21/24) of the patient population recognized Pla l 2. The allergenic relatedness between Pla l 2, Pho d 2 and six pollen profilins was confirmed, and IgE cross-reactivity of Pla l 2 with rBet v 2 and rPhl p 12 was demonstrated. Pla l 2 is the profilin from P. lanceolata. The demonstrated allergenicity of this protein and its cross-reactivity with other pollen profilins support its use in profilin diagnostic assays.

Tromboembolismo pulmonar bilateral masivo puerperal, secundario a trombosis venosa profunda intraparto. A propósito de un caso / Postpartum massive bilateral pulmonary thromboembolism secondary to deep intrapartum venous thrombosis. A case report

Resumen ANTECEDENTES: la enfermedad tromboembólica es un cuadro clínico que puede manifestarse durante el embarazo, como un episodio aislado de trombosis venosa profunda en los miembros inferiores, o como tromboembolismo pulmonar. La prevalencia se considera baja: 1 caso por cada 1600 embarazos. El solapamiento de los síntomas de la enfermedad tromboembólica con el embarazo puede retrasar la sospecha clínica y complicar el establecimiento del diagnóstico. CASO CLÍNICO: paciente primigesta, de 37 años de edad, con un episodio de tromboembolismo pulmonar con inicio en el puerperio, asociado con trombosis venosa profunda intraparto como factor desencadenante que evolucionó favorablemente gracias al diagnóstico y tratamiento oportunos. CONCLUSIÓN: la importancia de este caso radica en la necesidad de actuar con rapidez para establecer el diagnóstico e indicar inmediatamente el tratamiento.

Abstract BACKGROUND: Venous thromboembolism (VTE) is a combination of pathologies, which can manifest during pregnancy as an isolated lower extremity deep vein thrombosis (DVT) or as a pulmonary embolus (PE). The global prevalence is low, which occurs in 1 to 1600 pregnancies. CLINICAL CASE: A 37-year-old primigravid patient with an episode of pulmonary thromboembolism with onset in the puerperium associated with intrapartum deep venous thrombosis as a triggering factor that evolved favorably due to timely diagnosis and treatment. CONCLUSION: The importance of this case lies in the need to act quickly to establish the diagnosis and immediately indicate the treatment.

Relevance of Allergenic Sensitization to Cynodon dactylon and Phragmites communis: Cross-reactivity With Pooideae Grasses.

BACKGROUND AND OBJECTIVES: The homologous group of sweet grasses belongs to the Pooideae subfamily, but grass pollen species from other subfamilies can also cause allergy, such as Cynodon dactylon (Chloridoideae) and Phragmites communis (Arundinoideae). C dactylon and P communis have not been included in the sweet grasses homologous group because of their low cross-reactivity with other grasses. The aims of this study were to investigate the profile of sensitization to C dactylon and P communis in patients sensitized to grasses and to analyze cross-reactivity between these 2 species and temperate grasses. METHODS: Patients were skin prick tested with a grass mixture (GM). Specific IgE to GM, C dactylon, P communis, Cyn d 1, and Phl p 1 was measured by ImmunoCAP. A pool of sera was used for the immunoblot assays. Cross-reactivity was studied by ELISA and immunoblot inhibition. RESULTS: Thirty patients had sIgE to GM. Twenty-four (80%) had positive results for C dactylon, 27 (90%) for P communis, 22 (73.3%) for nCyn d 1, and 92.9% for rPhl p 1. Bands were detected in the 3 extracts by immunoblot. Inhibition of GM was not observed with C dactylon or P communis by immunoblot or ELISA inhibition. When C dactylon or P communis were used in the solid phase, GM produced almost complete inhibition. CONCLUSIONS: Eighty percent of patients sensitized to grasses were also sensitized to C dactylon and 90% were sensitized to P communis. Sensitization to these species seems to be induced by allergens different to those in sweet grasses.

In vivo diagnosis with purified tropomyosin in mite and shellfish allergic patients.

BACKGROUND: Tropomyosin is the most studied shellfish allergen and has been involved in cross-reactivity among different invertebrates (crustacean, mollusks, mites, insects, and nematodes). OBJECTIVE: To determine the relevance of tropomyosin in mite- and shellfish-sensitized patients using tropomyosin skin testing. METHODS: Patients were divided into 3 groups: group M included mite allergic patients (ie, individuals with respiratory symptoms and a positive result on skin prick testing [SPT] to house dust mites), group S included shellfish allergic patients (ie, individuals who reported symptoms with shellfish), and group MS included mite- and shellfish allergic patients (ie, individuals who simultaneously fulfilled the inclusion criteria for groups M and S). Tropomyosin was purified from shrimp, characterized, and used in SPT for diagnosis in the patient population. RESULTS: Eight hundred fifty patients were included in the study: 790 (92.9%) in group M, 21 (2.5%) in group S, and 39 (4.6%) in group MS. Tropomyosin was purified from shrimp with a purity higher than 95%. Forty-two individuals tested positive to tropomyosin: the prevalence was 2.7% in group M, 28.6% in group S, and 38.5% in patients of group MS. Twenty-one (50%) of the tropomyosin-positive individuals had symptoms with shellfish, and 3 (14.3%) reported anaphylaxis. CONCLUSION: The prevalence of tropomyosin was low in mite-sensitized patients (2.7 %) and high in shellfish allergic patients (28.6%). The higher prevalence of tropomyosin was found in patients sensitized to both mite and shellfish (38.5%). The selection of tropomyosin-sensitized patients by SPT might help in the choice of appropriate treatments or management for these patients.

Cloning and characterization of tropomyosin from the mite Chortoglyphus arcuatus.

Tropomyosin is a pan-allergen that shares a high homology among species. It is involved in cross-reactivity among mites, crustaceans, mollusks and insects. The objectives were to express and purify recombinant tropomyosin from the storage mite Chortoglyphus arcuatus, and to investigate the homology and cross-reactivity with tropomyosin from other invertebrates. Recombinant C. arcuatus tropomyosin (r-Cho a 10) was selected from a library by screening with a pool of patient sera. r-Cho a 10 (UniProt: H2DFL1) was sequenced, expressed in Escherichia coli and purified by ion exchange and affinity chromatography. Polyclonal anti-tropomyosin antibodies were produced in mice. IgE recognition of r-Cho a 10 was checked by immunoblot. Immunoblot inhibition assays were used to identify the native tropomyosin in the complete extract from C. arcuatus and study cross-reactivity between r-Cho a 10 and Der p 10. Identification of tropomyosin in other allergenic sources was performed by immunoblot. r-Cho a 10 showed a high homology (54-96%) with other tropomyosins from different allergenic sources. IgE recognition was observed using a pool of sera from sensitized individuals. Tropomyosins from different extracts were identified not only in the whole C. arcuatus extract but also in Dermatophagoides pteronyssinus, shrimp, mussel, cockroach and Anisakis extracts with polyclonal α-Cho a 10. r-Cho a 10 completely inhibited the recognition of Der p 10. Recombinant C. arcuatus tropomyosin maintained its capacity to recognize IgE. r-Cho a 10 was used to prove cross-reactivity among tropomyosins from other invertebrate species, including mites. This is the first C. arcuatus allergen included in the WHO/IUIS (World Health Organization/International Union of Immunological Societies) Allergen Nomenclature database.