Effect of the chicken zp1 gene on osteoblast mineralization / 生物工程学报

The aim of this study was to clone the chicken zp1 gene encoding zona pellucida 1 (Zp1) and investigate its tissues expression profile and its effect on osteoblast mineralization. The expression level of zp1 was quantified in various tissues of laying hens and in the tibia of the pre- and post-sexual maturity by RT-qPCR. Zp1 overexpressed vector was transfected into chicken calvarial osteoblasts which were induced differentiation for 8 days, and the extracellular mineral and the expression of mineralization-related genes were detected. The full-length chicken zp1 gene is 3 045 bp, encoding 958 amino acids residuals, and has two N-glycosylation sites. The highest expression level of the zp1 gene was found in the liver, followed by the tibia and yolk membrane, while no expression was detected in the heart and eggshell gland. Compared with the pre-sexual maturity hens, the concentration of estrogen (E2) in plasma, the content of glycosaminoglycan (GAG) and the expression level of the zp1 gene in the tibia with post-sexual maturity were higher. The extracellular matrix and the level of osteoblast mineralization-related genes showed a significantly upregulated expression in chicken calvarial osteoblasts with Zp1 overexpressed and addition of estrogen. The expression of the zp1 gene is tissue-specific and positively regulated osteoblast mineralization under the action of estrogen, laying the foundation for elucidating the functional properties of Zp1 in chicken bones during the egg production period.

Segurança da vacina de febre amarela em pacientes comprovadamente alérgicos à proteína do ovo / Safety of yellow fever vaccine in egg allergic patients

Introdução: A vacina de febre amarela, recomendada em áreas endêmicas, é contraindicada em alérgicos à proteína do ovo (APO) por ser cultivada em ovos de galinha embrionados. Objetivo: O objetivo do estudo foi mostrar a segurança da vacina de febre amarela em pacientes comprovadamente APO. Método: Foi realizado estudo prospectivo em hospital quaternário, no período de janeiro a outubro de 2018. Foram incluídos pacientes com APO confirmada por teste de provocação oral (TPO), reação anafilática à proteína do ovo nos últimos 6 meses, ou reação de APO nos últimos 2 meses associada à IgE específica positiva. Todos foram submetidos ao teste de puntura com a vacina na apresentação pura. Se negativo, realizado teste intradérmico (ID) com a vacina na diluição de 1:100. Se ID negativo, vacina aplicada em dose plena. Se teste de puntura ou ID positivo, vacina aplicada fracionada segundo protocolo de dessensibilização. Resultados: Dos 78 pacientes com história presumida de APO, confirmou-se o diagnóstico em 43 (30M:13F, mediana idade 2,7 a): 30 por TPO, 7 com anafilaxia em menos de 6 meses da vacina, e 6 com reação imediata após ingestão do ovo há menos de 2 meses e IgE específica positiva. Durante o TPO, 12 apresentaram anafilaxia, e os demais (18) apresentaram urticária e/ou angioedema ou vômitos. Todos os testes de puntura (43) foram negativos. ID foi negativo em 37 pacientes, que receberam a dose plena da vacina, sem reações. Apenas 6 apresentaram ID positivo e necessitaram dessensibilização para vacina. Metade desses pacientes (3/6) apresentou reações de hipersensibilidade leves e foi tratada com anti-H1 e/ou corticoide oral. O ID positivo foi significativamente relacionado à reação à vacina (p = 0,0016). Conclusão: Concluiuse ser possível vacinar alérgicos a ovo, com um protocolo seguro, mesmo em paciente comprovadamente anafilático. É necessária uma unidade especializada para sua realização, com capacidade de controlar possíveis situações de risco.

Introduction: The yellow fever vaccine (YFV) is recommended in endemic areas, but represents a risk for egg allergic (EA) patients, as it is cultivated in chicken embryos. Objective: This study aimed to describe the outcomes of YFV in patients with confirmed egg allergy. Methods: A prospective study was conducted in a quaternary hospital, from January to October 2018. EA was diagnosed through oral food challenge (OFC) or recent history of anaphylaxis following egg contact in the past 6 months or allergic reaction in the past 2 months with positive specific immunoglobulin E (IgE). Skin prick testing (SPT) with YFV was performed in all participants. If SPT was negative, an intradermal test (IDT) was performed at 1:100 dilution. If IDT was negative, a full dose of YFV was administered. If SPT was positive, the YFV was administered using a graded-dose protocol. Results: Among 78 patients with prior history of EA, 43 were confirmed (30 male to 13 female, median age of 2.7 years). Thirty patients had a positive OFC, seven reported recent anaphylaxis, and six had reactions in the past 2 months with positive specific IgE. During OFC, 12 patients had anaphylaxis and 18 had urticaria and/or angioedema or vomiting. SPT with YFV was negative in all patients (43). IDT was negative in 37 patients, who received a full dose of YFV, uneventfully. Six patients had a positive IDT and received the YFV in graded doses; half of them had a mild reaction controlled with antihistamines and three patients received the vaccine without reactions. Positive IDT was significantly related to vaccine reaction (p=0.0016). Conclusion: The YFV using a specific protocol was safe even in anaphylactic patients. An appropriate setting is required in order to control possible adverse events.

Preparation and characterization of rabbit anti-mouse zona pellucida 2 antibodies / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare rabbit anti-mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their immunoactivity.</p><p><b>METHODS</b>Recombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by sperm-egg binding assay.</p><p><b>RESULTS</b>ELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted sperm by 43.7%.</p><p><b>CONCLUSION</b>We obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes with acrosomal reacted sperm.</p>

Expression of Ki-67 and estrogen receptor in the uterus of mice with autoimmune premature ovarian failure induced by peptide zona pellucida 3 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the histomorphology and the expressions of the proliferation marker Ki-67 and estrogen receptor in the uterus of mice with autoimmune premature ovarian failure (POF) induced by zona pellucida 3 peptide (pZP3).</p><p><b>METHODS</b>Autoimmune POP models were established in 20 female BALB/c mice (7-8 weeks old) by immunization with pZP3 and another 20 mice served as the control group. The POP models were verified by vaginal cytology, serum sex hormones, ovary histomorphology and ZP3 antibody immunohistochemistry. The histomorphology and expressions of Ki-67, estrogen receptor α and estrogen receptor β in the uterus of the mice were detected.</p><p><b>RESULTS</b>Autoimmune POP models were established successfully in 80% of the mice at 8 weeks after the immunization. Compared with those in the control group, the mice in the model group showed a smaller volume of the uterus, thinner endometrium and a reduced number of glands. The luminal epithelial cells, glandular epithelial cells and stromal cells in the uterus of the model mice all presented with a lower expression of Ki-67 than those in the control group, and Ki-67 translocation from the nuclei to the cytoplasm was found in the model group. The luminal epithelial cells, glandular epithelial cells and stromal cells showed positive ERα immunoreactivity in the model group but not in the control group. No obvious ERβ expression was found in the uterus in either of the groups.</p><p><b>CONCLUSION</b>pZP3 can induce autoimmune POP, cause suppressed proliferation of the endometrial epithelial cells and stromal cells, and reduce the cellular expression of ERα in the uterus of mice.</p>

Diagnostic utility of skin prick test to cooked egg in children with egg allergy

PURPOSE: Reliable predictors of tolerance to cooked egg in an egg allergic population are not established. We investigated the usefulness of the skin prick test to cooked egg in children with egg allergy. METHODS: We studied 36 children with egg allergy. Skin prick tests (SPTs) for the uncooked or cooked form of egg white and egg yolk, whole egg, ovomucoid (OVM), and ovalbumin (OVA) were performed at diagnosis. The reagents of cooked egg for SPT were prepared by baking for 25 minutes in 200 degree oven. We also examined specific IgE levels to whole egg, egg white, egg yolk, OVM, and OVA. RESULTS: Patients with history of allergic reaction to extensively heated egg showed significantly increased wheal size for cooked egg white (median [interquartile range]), 10.5 [7.0-14.6] vs. 4.2 [0.0-5.6], P<0.001) and OVM (9.6 [7.3-13.8] vs. 5.6 [0.0-7.8], P=0.001) than those without the history. The strongest positive correlation was found between wheal size for cooked egg white and OVM (r=0.788, P<0.001). SPT wheal size for cooked egg white were positively correlated with serum OVM-specific IgE levels (r=0.691, P<0.001). Cutoff value was 7.0 mm in SPT wheal size for cooked egg white, the sensitivity was 73.1% and specificity was 99.0%. SPT for cooked egg white showed significantly higher area under curve than serum egg white specific IgE. CONCLUSION: Our results suggest that SPT to cooked egg white may be useful predictor of allergic reaction to cooked egg. Further investigations will be needed.

Listeriosis en México: importancia clínica y epidemiológica / Listeriosis in Mexico: Clinical and epidemiological importance

La listeriosis es una enfermedad transmitida por alimentos (ETA) y ocasionada por Listeria monocytogenes. La importancia de ésta se debe a su impacto clínico, la alta tasa de mortalidad y el efecto económico derivado de los brotes asociados con el consumo de alimentos. En México, las fallas en los sistemas de vigilancia epidemiológicos son causa de información imprecisa sobre la incidencia de la listeriosis y sobre su caracterización como ETA. En este trabajo se presentan datos referentes a la presencia de la bacteria en alimentos, reportes de casos de la enfermedad y patologías relacionadas con infección por L. monocytogenes. La falta de datos exactos sobre la importancia de esta bacteria plantea la necesidad de concientizar a las instancias correspondientes para definir estrategias de búsqueda intencionada de L. monocytogenes en alimentos y de la recopilación de información clínica precisa que permita conocer la importancia clínica y epidemiológica de la listeriosis en México.

Listeriosis is caused by Listeria monocytogenes, an important food-borne disease due to its clinical forms, high mortality rate, and the economic impact in both clinical and food production industries. In Mexico, the lack of epidemiological surveillance systems leads to the need of accurate data on the incidence of listeriosis and its association with food-borne disease. In this paper, we present data about the presence of this bacterium in food, reports related to clinical cases of listeriosis, and information of diseases in which L. monocytogenes may be involved. However, in most of these cases the etiology was not established. Given this, there's a need to inform and warn the appropriate entities, to define strategies for the mandatory search of L. monocytogenes through the whole food production chain and clinical suspects, for the epidemiological importance and control of listeriosis in Mexico.

Expression of recombinant human ZP3 protein using the baculovirus expression system / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the methods and solve the technical bottlenecks in the preparation of recombinant human protein hZP3 using the baculovirus expression system and pave the technical ground for the production and application of recombinant hZP3.</p><p><b>METHODS</b>The recombinant vector pFASTBAC HTa-hZP3 was constructed and transferred to competent E. coli cells carrying bacmid to produce recombinant bacmid by homologous recombination. Sf9 cells were transfected with the recombinant bacmid to produce recombinant baculovirus. Full-length recombinant hZP3 (amino acids 1-424) and truncated recombinant hZP3 (amino acids 23-348) were expressed in the sf9 cells by infection with the recombinant baculovirus. The expression time of hZP3 was determined by Western blot and its purification was explored.</p><p><b>RESULTS</b>The recombinant bacmid and baculovirus were successfully constructed for expressing both the full-length and truncated hZP3. The maximal expression of recombinant hZP3 in the sf9 cells was achieved at 72-96 hours after baculovirus infection. Some of the recombinant hZP3 with His-tag could bind affinity matrix and got purified but most of the solubilized hZP3 passed through and the reasons remained unknown. Purified recombinant hZP3 labeled with Dylight Dye488 was able to bind human sperm.</p><p><b>CONCLUSION</b>It is feasible to express recombinant hZP3 in insect cells using the baculovirus system though the yield of hZP3 needs to be optimized. The methods for efficient enrichment and purification of recombinant hZP3 require further exploration.</p>

Effect of different levels of synbiotic on productivity and egg quality, blood parameters and hatchability in broiler breeder hens

The present study deals with the effect of different levels of synbiotic on egg production and quality, blood parameters, hatchability and yolk cholesterol in broiler breeder hens. The aim of this study was to determine the best level of symbiotic in the diet of broiler breeder hens. The study was conducted based on a completely randomized design with 4 treatment groups, 2 replicates and 10 controls in each experimental unit. The experimental rations were containing 0 [control S0], 0.05 [S0.05], 0.1 [S0.1] and 0.2% [S0.2] synbiotic. Egg quality and weight were measured daily. Blood parameters, laying rate and hatchability were measured weekly. Yolk cholesterol, ovarian weight and number of ovarian large follicles were evaluated at the end of the experiment. The data were analyzed using GLM procedure. The results showed that yolk index in group S0.1 was significantly lower [p<0.05] than groups S0.05 and S0.2, but was not statistically different from the control [p>0.05]. Also, synbiotic had no significant effect on other egg quality traits, egg weight and hatchability [p>0.05]. Addingy synbiotic supplement to diet reduced significantly the laying rate, ovarian weight and number of ovarian large follicles in the group S0.2 compared with the other groups [p<0.05]. Synbiotic significantly increased plasma glucose concentration [p<0.05], but plasma triglyceride, HDL and yolk cholesterol concentrations were not influenced by dietary symbiotic [p>0.05]. Furthermore, plasma cholesterol concentration [last week] in groups S0.1 and S0.2 were significantly lower than control group [p<0.05]. The results showed that it would be proper to use 0.1% symbiotic in the diet of broiler breeder hens

Efeito dos níveis de energia para poedeiras comerciais no período final de produção sobre o desempenho, a conversão alimentar e energética e a qualidade de ovos / Effect of energy levels for old layers on performance, feed and energy conversion ratios and egg quality

Foram utilizadas 1200 poedeiras comerciais da linhagem Lohmann LSL, com 64 semanas, em gaiolas convencionais. Estas foram divididas em cinco grupos, de acordo com o nível de energia metabolizável aparente corrigida pelo nitrogênio (EMAn) presente em sua dieta: 2700kcal/kg; 2775kcal/kg; 2850kcal/kg; 2925kcal/kg; e 3000kcal/kg, com seis repetições. As aves receberam as rações, á base de milho e farelo de soja, por 16 semanas. O delineamento experimental foi inteiramente ao acaso, e as médias foram submetidas à regressão polinomial e ao teste de Kruskal-Wallis. O aumento de EMAn não exerceu efeito sobre a produção, a massa e o peso de ovos, bem como sobre o peso das aves e a viabilidade. O consumo de ração diminuiu de acordo com o aumento na energia da dieta, porém a conversão energética foi pior com esse aumento. Em contrapartida, a conversão alimentar melhorou o aumento dos níveis de EMAn. O consumo de EMAn é constante independentemente do seu nível dietético. Não há diferença nas unidades Haugh, nos sólidos da gema e na porcentagem dos componentes do ovo. O peso específico melhora com o aumento dos níveis de EMAn. Os níveis nutricionais para poedeiras velhas podem estar sendo superestimados.

A total of 1200 Lohmann LSL laying hens, with 64 weeks of age in conventional cages was used. These were divided into five groups according to the level of apparent metabolizable energy corrected for nitrogen retention (AMEn) present in the diet: 2700kcal/kg; 2775kcal/kg; 2850kcal/kg; 2925kcal/kg and 3000kcal/kg, with six replicates. The birds received the diets based on corn and soybean meal for 16 weeks. The experimental design was completely randomized and the means were submitted to polynomial regression and the Kruskal-Wallis test. The increased level of AMEn had no effect on production, egg mass and weight. There was no influence of AMEn on bird weight, egg weight and viability. Feed intake decreased with increasing levels, however, and the conversion of AMEn was worse with its increase, and in return feed conversion worsened with increasing consumption. AMEn is constant regardless of the level of the feed. There is no difference in Haugh units, yolk solids and percentage of egg components. The specific gravity improves with increasing levels of AMEn. The nutritional levels for older laying hens may be overestimated.

Therapeutic effect of Bushen Huoxue recipe on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and mechanism of Bushen Huoxue Recipe (, BHR) in the treatment of murine autoimmune premature ovarian failure (POF).</p><p><b>METHODS</b>The recombinant porcine zona pellucida 4 (pZP4) was expressed in E. coli BL21 (DE3) strain within prokaryotic plasmid pET28a (+), purified by Ni-affinity chromatography and verified by Western blot. Murine autoimmune POF model was established by immunization with pZP4 of female BALB/c mice. Fifty POF mice were randomly divided into 5 groups, which were respectively given low (3.75 mg/kg), moderate (7.5 mg/kg), and high dose (15.0 mg/kg) of BHR by gastrogavage once daily for 20 days, with 17-β-estradiol (0.13 mg/kg) and normal saline as positive and negative control. Estrous cycles were analyzed through vaginal smears, serum estradiol (E) levels, and anti-pZP4 antibody titers were detected by ELISA. The proliferative responses in vitro of spleen lymphocytes to pZP4 antigen restimulation were measured by [(3)H]-thymidine incorporation, and the histomorphology changes of ovary were evaluated by optical microscope.</p><p><b>RESULTS</b>The purified pZP4 was visible as a single lane with 14.4 kD in SDS-PAGE and Western blot. The murine POF model with lengthening estrous cycles, decreased levels of serum E2, high titers of serum anti-pZP4 antibody, and reduced ovarian follicles and corpus lutea were established by immunization with recombinant pZP4. Treatment with moderate and high dosage BHR significantly increased ovarian follicles and reduced the proliferation of spleen lymphocytes to the pZP4 antigen of POF mice (P <0.05). However, only the high dosage BHR administration significantly improved the estrous cycles, elevated the serum E levels (P <0.01), and decreased the serum anti-pZP4 antibody titers of model mice P<0.05).</p><p><b>CONCLUSIONS</b>The recombinant pZP4 could evoke the antigen-specific immune response in mice and induce the autoimmune ovarian injury. It has been demonstrated that BHR was able to increase the serum E levels and protect ovarian functions from the autoimmune injury in murine POF model.</p>

The Clinical Usefulness of IgE Antibodies Against Egg White and Its Components in Korean Children

PURPOSE: Egg (egg white) allergies are among the most common food allergies in infants and young children. Serum egg white-specific IgE (sIgE) levels have been shown to be correlated with clinical symptoms, and the predictive decision point of sIgE levels has been proposed and used widely in the clinical setting. However, some patients whose sIgE levels to egg white are higher than the predictive decision point value show no clinical symptoms, and vice versa. This study was conducted to evaluate the clinical usefulness of sIgE antibodies to egg white and its components in the diagnosis of egg allergies. METHODS: Forty-one patients younger than 2 years of age with no experience of egg intake due to concerns regarding allergies or a non-specific clinical response to eggs were enrolled. Total IgE levels and the levels of IgE antibodies specific for egg white and its components (ovomucoid, ovalbumin, and conalbumin) were measured by ImmunoCAP testing. The clinical response of the subjects was confirmed by an open oral food challenge (OFC). RESULTS: Fifteen (71.4%) out of 21 patients in the egg white-sIgE > or =2 kU/L group showed a positive response, while 10 (50.0%) out of 20 patients in the egg white-sIgE <2 kU/L group showed a negative response to the OFC. There were no statistically significant differences in the levels of sIgE antibodies against egg white and its components between the positive and negative open OFC groups. In addition, there were no statistically significant differences in the levels of sIgE antibodies against egg white and its components based on an intra-group analysis. CONCLUSIONS: Our results show that the sensitivity and specificity of the predictive decision point values for egg white-sIgE antibodies by ImmunoCAP were relatively low in Korean children. In addition, no egg white component predicted the clinical reactivity of the subjects. We suggest that the predictive decision point value for a positive egg oral challenge test by ImmunoCAP should be re-evaluated. Moreover, we suggest that careful personal history recording and challenge tests are necessary for the correct diagnosis of an egg allergy.

The Influence of the Time and Temperature of Heat Treatment on the Allergenicity of Egg White Proteins

PURPOSE: The present study was performed to determine the factor, either duration or the temperature of heat treatment, exerting maximal and significant influence on the composition and allergenicity of egg white (EW) proteins. METHODS: Raw EW and 4 kinds of heated EW (fried EW, boiled EW for 10 minutes, boiled EW for 30 minutes, and baked EW for 20 minutes at 170degrees C) were prepared, and subsequently protein extraction was carried out. The proteins were separated by SDS-PAGE, and then immunoglobulin E (IgE) immunoblots were performed with the sera of 7 egg-allergic patients. Furthermore, the antigenic activities of ovalbumin (OVA), ovomucoid (OM), and ovotransferrin (OT) in different EW samples were measured by inhibition enzyme-linked Immuno-sorbent assay (ELISA). RESULTS: In SDS-PAGE analysis, the intensity of the protein band at 45 kD (corresponding to OVA) decreased significantly in boiled EW (30 minutes) and baked EW, but no change was observed in the case of boiled EW for 10 minutes. In IgE immunoblots, the IgE response to 34-50 kD (OM and OVA) in boiled EW for 30 minutes decreased significantly, when compared with raw EW and other heated EWs. In inhibition ELISA, a significant decrease in the OVA antigenic activity was observed in boiled EW for 30 minutes amongst other heated EW samples. However, OM antigenic activity in all kinds of heated EW including boiled EW for 30 minutes did not reduce after heat treatment. The OT antigenic activity nearly disappeared in heated EWs except in the case of boiled EW for 10 minutes. CONCLUSIONS: Amongst 4 kinds of heated EWs, the boiled EW for 30 minutes showed the most significant changes both in composition and reduction in allergenicity. Our results revealed that the duration of heat treatment had more influence on the composition and allergenicity of EW proteins than the temperature.

The Influence of the Presence of Wheat Flour on the Antigenic Activities of Egg White Proteins

PURPOSE: It is known that ovomucoid, an egg allergen, is heat resistant and remains soluble after heating. However, a recent study showed that the antigenic activity of ovomucoid could be reduced by heating when egg white (EW) was mixed with wheat flour. This study was performed to determine the influence of wheat flour on the antigenic activities of EW proteins when EW is heated, and the influence of the duration of heat treatment. METHODS: A mixture of EW and wheat flour was kneaded for 10 minutes and then baked at 180degrees C for 10 minutes and 30 minutes. The EW without wheat flour was also heated at 180degrees C for 10 minutes and 30 minutes. The proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and IgE immunoblotting was performed with the pooled sera of 5 egg-allergic patients. The antigenic activities of ovomucoid in different EW samples were measured by inhibition enzyme-linked immunosorbent assay (ELISA). RESULTS: 1) SDS-PAGE: the intensity of the 37-50 kD bands (overlapped bands of ovomucoid and ovalbumin) decreased significantly in the mixture of EW and wheat flour baked for 30 minutes, compared with the mixture baked for 10 minutes, heated EW and raw EW. 2) IgE immunoblot: in the mixture of EW and wheat, a remarkable decrease of IgE reactivity to 37-50 kD was observed when baked for 30 minutes. 3) Inhibition ELISA: the antigenic activity of ovomucoid decreased significantly in the mixture of EW and wheat baked for 30 minutes, but not in the heated pure EW. CONCLUSIONS: This study showed that the antigenic activity of ovomucoid can be reduced by baking EW with wheat flour. The decrease in ovomucoid antigenicity in the baked mixture of EW and wheat flour was dependent on the time of heat treatment, indicating that heating should be prolonged to achieve a reduction in ovomucoid antigenic activity.

Alergia al huevo en Pediatría. Documento del Comité de Pediatría de la Asociación Argentina de Alergia e Inmunología Clínica / Egg allergy in pediatrics. Pediatric Committee document of the Argentina Association of Allergy and Clinical Immunology

El huevo de gallina es una fuente de proteínas de alto valor biológico de bajo costo y de vitaminas del complejo B, importantes para la alimentación del niño. Culturalmente es uno de los alimentos básicos de nuestra dieta y, debido a esto, la alergia a sus proteínas es una de las más frecuentes en la infancia y tiene su mayor impacto en niños en edad preescolar. Estos niños representan una población vulnerable debido a que se encuentran en una etapa importante de su crecimiento y desarrollo, y el tratamiento de esta patología genera la adopción de dietas restrictivas que pueden impactar en forma negativa en su salud y calidad de vida. Este impacto está dado en parte por la ubicuidad de sus proteínas, que limita ampliamente la dieta y genera riesgos de reacciones alérgicas que se incrementan a medida que el niño crece y alcanza una mayor independencia. Teniendo en cuenta la importancia de esta patología, el Comité de Pediatría realizó una revisión actualizada con el fin de proveer herramientas útiles para el manejo adecuado de la misma. (AU)

Eggs are a source of low cost high biological value protein and complex B vitamins important for the child's nutrition. Culturally it is one of the staples of our diet and because of this, egg allergy is one of the most common food allergies in childhood and has its greatest impact on preschool children. These children represent a vulnerable population because they are at an important stage in their growth and development and the treatment of this condition generates the adoption of restrictive diets that may impact negatively on their health and quality of life. This impact is given in part by the ubiquity of their proteins that largely restrict the diet and generates risks of allergic reactions that increase as the child grows and earns greater independence. Given the importance of this issue the Pediatrics Committee conducted an updated review to provide useful tools to manage this condition.(AU)

Changes of prostaglandin D2,carboxypeptidase A3 and platelet activating factor in guinea pig in anaphylactic shock / 法医学杂志

OBJECTIVE@#To detect the changes of leukotriene E4(LTE4), prostaglandin D2(PGD2), carboxypeptidase A3(CPA3) and platelet activating factor (PAF) in guinea pigs died from anaphylactic shock.@*METHODS@#Guinea pigs were used for establishing anaphylactic shock models. The levels of LTE4, PGD2 and CPA3, and PAF were detected in urine, plasma, and brain tissues with ELISA kit, respectively. The significant biomarkers were selected comparing with control group. The changes of PGD2, CPA3 and PAF in the guinea pigs at time zero, 12 and 24 hours after death were observed and compared respectively. The effect of platelet activating factor acetylhydrolase (PAF-AH) to PAF in guinea pig brain was examined and compared.@*RESULTS@#There were no statistically differences of LTE4 levels in urine observed between experimental group and control group. The levels of CPA3, PGD2 and PAF in the experimental group were significantly higher than that in the control group at 0 h. The levels of PAF at 12 and 24 hours after anaphylactic shock were significantly higher than that in the control group. The levels of PAF decreased significantly after pretreatment with PAF-AH.@*CONCLUSION@#LTE4 in urine cannot be selected as a biomarker to determine the anaphylactic shock. PGD2 and CPA3 in plasma, and PAF in brain tissue may be used as biomarkers to determine the anaphylactic shock. PAF-AH may be potentially useful for clinical treatment of anaphylactic shock.

Immunostimulatory property of a synthetic peptide belonging to the soluble ATP diphosphohydro-lase isoform (SmATPDase 2) and immunolocalisation of this protein in the Schistosoma mansoni egg

A peptide (SmB2LJ; r175-194) that belongs to a conserved domain from Schistosoma mansoni SmATPDase 2 and is shared with potato apyrase, as predicted by in silico analysis as antigenic, was synthesised and its immunostimulatory property was analysed. When inoculated in BALB/c mice, this peptide induced high levels of SmB2LJ-specific IgG1 and IgG2a subtypes, as detected by enzyme linked immunosorbent assay. In addition, dot blots were found to be positive for immune sera against potato apyrase and SmB2LJ. These results suggest that the conserved domain r175-194 from the S. mansoni SmATPDase 2 is antigenic. Western blots were performed and the anti-SmB2LJ antibody recognised in adult worm (soluble worm antigen preparation) or soluble egg antigen antigenic preparations two bands of approximately 63 and 55 kDa, molecular masses similar to those predicted for adult worm SmATPDase 2. This finding strongly suggests the expression of this same isoform in S. mansoni eggs. To assess localisation of SmATPDase 2, confocal fluorescence microscopy was performed using cryostat sections of infected mouse liver and polyclonal antiserum against SmB2LJ. Positive reactions were identified on the external surface from the miracidium in von Lichtenberg's envelope and, in the outer side of the egg-shell, showing that this soluble isoform is secreted from the S. mansoni eggs.

Gamete interactions in teleost fish: the egg envelope. Basic studies and perspectives as environmental biomonitor

The current knowledge about teleost fish egg envelope is summarized. The paper analyzes the organization and deposition process of the protein composition and genes involved in the synthesis of teleost fish egg envelopes and their role in gamete interaction during fertilization. Pelagic and demersal species that our research group is working with are especially considered. The vertebrate ZP family of proteins, the evolution and relationship among the different genes and their expression are taken into account. We consider fish envelope as a possible biomonitor for ecological contaminants. The biotechnological applications for aquaculture and genomic and post-genomic approaches are auspicious.

Binding characteristics of sperm with recombinant human zona pellucida glycoprotein-3 coated beads.

Background & objectives: An inability or decreased ability of spermatozoa to bind to the zona pellucida (ZP), an extracellular glycoproteinaceous matrix surrounding egg, is one of the plausible causes of idiopathic infertility. It will be clinically useful to distinguish this condition from other causes of infertility. An assay system, investigating binding of human sperm with ZP glycoprotein may prove useful in this regard. We attempted to develop a simple assay system to analyse the binding of capacitated human spermatozoa to human zona pellucida glycoprotein-3 (ZP3) using baculovirus-expressed recombinant human ZP3 coated beads. Methods: Recombinant baculovirus-expressed ZP3 was purified, labelled with biotin and coated on streptavidin sepharose beads. An in vitro assay system was optimized to study binding of capacitated human sperm to ZP3 coated beads. Results: A higher percentage of baculovirus-expressed recombinant human ZP3 coated beads showed significant (P<0.05) binding of capacitated human sperm as compared to beads coated with fetuin. An inhibition in the binding of sperm to ZP3 coated beads was observed in presence of cold recombinant human ZP3. Further, prior incubation of ZP3 coated beads with monoclonal antibodies (MAbs) against ZP3 but not against ZP2 resulted in the decrease in number of sperm bound to bead. Interpretation & conclusion: An in vitro assay system to study the binding of human sperm to ZP3- primary sperm receptor was established, which may be useful to determine the functional competence of spermatozoa.

Soluble expression, purification and immunoreactive identification of mouse zona pellucida 3 fusion protein / 生物工程学报

Mammalian zona pellucida 3(ZP3) plays an important role in the induction of capacitating sperm acrosome reaction. In this study, we obtained the soluble mZP3 fusion protein and identified its immunoreactivity. mZP3 cDNA was cloned into plasmid pMAL-p2x, and the recombinant plasmid was transformed into Escherichia coli BL21. To get the soluble mZP3 fusion protein, we tried to optimize the expression conditions, including additives, IPTG concentrations, temperatures and induction duration. Then, Western blotting and ELISA were used to identify the immunoreactivity of the purified protein. Based on the optimization experiments, we concluded that the best soluble expression conditions for the mZP3 fusion protein involved incubation to an A600 of 0.6, addition of glucose to a final concentration of 0.02 mol/L, addition of IPTG to a final concentration of 0.6 mmol/L and then further incubation for 4 h at 25 degrees C. Western blotting and ELISA showed that the mZP3 fusion protein retained immunoreactivity. The fusion protein can be used as solubility antigens for developing the immunocontraception vaccines of mZP3 and detecting the immune effects of the vaccine.