Clinical characteristics of IgE-mediated cow's milk protein allergy in children / 中华儿科杂志

Objective: To analyze the clinical features of IgE-mediated cow's milk protein allergy (CMPA) in children aged 0-5 years. Methods: This cross-sectional study collected the data on children diagnosed with CMPA in the Department of Allergy at the Children's Hospital of the Capital Institute of Pediatrics from October 2019 to November 2020 and improved peripheral blood routine,total IgE defection, milk specific IgE (sIgE) defection,SPT and milk component defection,diagnosis of severe anaphylaxis based on clinical manifestations. Rank-sum test and chi-square test are used for statistical analysis of clinical characteristics between groups. Results: A total of 106 children (67 boys and 39 girls) were enrolled with the age of 15 (8, 34) months, including 42 cases (≤ 1 year of age), 39 cases (>1-<3 years of age) and 25 cases(≥3 years of age), the onset age of 6 (5, 8) months. Among them, 95 cases (89.6%) were reacted after consuming milk or its products, 42 cases (39.6%) had reaction due to skin contact and 11 cases (10.4%) reacted after exclusive breastfeeding. The onset time of milk product consumption was 45 (1, 120) min, skin contact pathway was 10 (5, 30) min and symptoms in breastfeeding pathway was 121 (61, 180) min. There was statistical difference among the time of symptoms (χ2=77.01, P<0.001).The cutaneous reaction was most common (100 cases, 94.3%), followed by digestive (20 cases, 18.9%) and respiratory (16 cases, 15.1%), and the nervous symptoms (1 case, 0.9%) were uncommon and 24 cases (22.6%) had at least one episode of anaphylaxis. There were 87 cases (82.1%) also diagnosed with other food allergies, 94 cases (88.7%) with previous eczema, 57 cases (53.8%) with history of rhinitis, and 23 cases (21.7%) with history of wheezing. The total IgE level was 191.01 (64.71, 506.80) kU/L, and the cow's milk sIgE level was 3.03 (1.11, 15.24) kU/L. The maximum diameter of the wheal in SPT was 8.2 (4.0, 12.0) mm. Component resolved diagnosis showed that 77 cases (81.9%) were sensitized to at least one out of 4 main components, including casein, α lactalbumin, β lactoglobulin and bovine serum albumin.The possibility of anaphylaxis in children with milk sIgE grade Ⅳ-Ⅵ was higher than that in children with grade 0-Ⅲ (57.7% (15/26) vs. 12.5% (10/80), OR=9.545, 95%CI 3.435-26.523). Children with milk SPT ≥+++ had a higher probability of anaphylaxis than those with milk SPT ≤++ (34.4% (11/32) vs. 11.5% (3/26), OR=4.016, 95%CI 0.983-16.400). Anaphylaxis were more common in α lactalbumin positive children than in negative children (34.3% (13/38) vs. 14.2% (8/56), χ2=1.23，P=0.042). Conclusions: CMPA in children has early onset and diversified clinical manifestations, which are mainly cutaneous symptoms. Most children are sensitized to at least one allergen component. Serum sIgE level, SPT reaction and allergen components play important roles in the diagnosis and evaluation of CMPA, and higher milk sIgE level may predict a higher risk of anaphylaxis.

Factor analysis as a tool to estimate association among individual proteins and other milk components with casein micelle size and cheese yield / Análise fatorial como ferramenta para estimar a associação entre proteínas individuais e outros componentes do leite com o tamanho das micelas de caseína e massa seca de queijo

The present study attempted to identify individual milk proteins and other milk components that are associated with casein micelle size (CMS) and dry matter cheese yield (DMCY) using factor analysis. Here, we used 140 bulk tank milk samples from different farms. Milk composition was determined using a Fourier transform infrared equipament. The individual milk proteins were (αS-casein, ß-casein, κ-casein, ß-lactoglobulin and α-lactoalbumin) measured by their electrophoretic profile. The CMS was estimated by photon correlation spectroscopy, and the DMCY was determined using reduced laboratory-scale cheese production. Factor analysis partitioned the milk components into three groups that, taken together, explain 68.3% of the total variance. The first factor was defined as "CMS", while the second as "DMCY" factor, based on their high loadings. The CMS was positively correlated with protein, casein, non-fat solids and αS-casein and negatively associated with κ-casein and ß-lactoglubulin. DMCY was positively correlated with fat, protein, casein, total solids and negatively correlated with αs-casein. These results indicate that the variation of individual milk proteins may be an important aspect correlated to milk quality and cheese production.(AU)

O objetivo do presente estudo foi avaliar a associação das frações proteicas individuais e de outros componentes do leite com o tamanho das micelas de caseína (TMC) e a produção de matéria seca de queijo (MSQ) utilizando-se análise fatorial. Foram coletadas 140 amostras de leite de tanque provenientes de diferentes fazendas. A determinação da composição do leite foi determinada por espectroscopia no infravermelho com transformação de Fourier. As proteínas individuais (αS-caseína, ß-caseína, κ-caseína, ß-lactoglobulina e α-lactalbumina) foram quantificadas pelo perfil eletroforético. O tamanho médio das micelas de caseína foi analisado pelo princípio de espectroscopia de correlação de fótons e pela produção MSQ a partir do modelo de coagulação do leite em escala reduzida. A análise fatorial delimitou as variáveis em três fatores, que, juntos, responderam por 68,3% da variação total dos dados. No primeiro fator foram observadas as associações mais fortes com o TMC, enquanto no segundo fator as correlações foram mais significativas com a MSQ. O TMC foi associado positivamente com o conteúdo de proteína, caseína, sólidos desengordurados e αS-caseína, e negativamente com κ-caseína e ß-lactoglubulina. MSQ foi associada positivamente com o teor gordura, proteína e caseína total, sólidos totais, e negativamente com o teor de αs-caseína. Esses resultados indicam que a variação quantitativa das proteínas do leite pode ser determinante da qualidade do leite na produção de queijo.(AU)

The effect of heat treatment or hydrolysis on cow's milk protein distributions and antigenicities

PURPOSE: Cow's milk protein is one of the most common and strongest food allergen. We investigated the effects of heat treatment on the distribution and antigenicities of major allergens from cow's milk. We also compared the protein distribution and antigenicities among cow's milk formula and its substitutes. METHODS: We heated alpha-casen, beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), and crude extract of cow's milk in 100degrees C boiling water for 1 hour. We prepared crude extracts from cow's milk formula, partially hydrolyzed milk formula (pHF) and extensively hydrolyzed milk formula (eHF). The protein compositions of all the samples were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigenicities were determined by IgE immunoblotting with pooled serum collected from 11 patients with milk allergy. RESULTS: After heating, no significant alteration was found in casein, and the aggregates of ALA and BLG were detected with molecular weights of about 30 and 45 kDa, respectively. The antigenicities of newly detected aggregates were increased. The new aggregates of BLG with increased antigenicities were also found in heated milk total protein. Major milk allergens were not found in pHF, and residual components with a molecular weight below 10 KDa did not show IgE-binding activity. We failed to observe the residual components and antigenicities of eHF. CONCLUSION: Changes in protein distribution and antigenicity of milk total protein induced by heat treatment may not be significantly different from those of each major allergen. The residual components of pHF could have little IgE-binding capacity, and there may be few or no antigenic components in eHF.

Successful management of severe cow's milk allergy with omalizumab treatment and CD-sens monitoring

Food allergy is common in children and young adults and may be difficult to diagnose and is at present treated with avoidance of the food in question. The aim of this report is to share our clinical experiences monitoring omalizumab treatment by basophil allergen threshold sensitivity, CD-sens. Five children, 6-16 years of age, with a severe milk allergy including episodes of anaphylaxis and IgE-antibodies, between 30 and 160 kU(A)/L to casein and alpha-lactalbumin (milk proteins), were treated with omalizumab. CD-sens, was tested prior to and after 4 months of omalizumab and if turned negative, it was followed by an oral milk challenge. All children became negative in CD-sens and had a negative milk challenge, but one child required doubling of the omalizumab dose to achieve a negative CD-sens before a challenge was done. Omalizumab appears useful in treatment of severe food allergy, e.g., anaphylaxis to milk, and CD-sens monitoring may decide when and how to perform a food challenge.

α-Lactoalbúmina como ingrediente de fórmulas infantiles

La α-lactoalbúmina es la principal proteína del lactosuero en la leche materna, alcanzando una concentración de 2,44 g/L en la leche madura. Su principal función es la síntesis de lactosa a partir de glucosa y galactosa en la glándula mamaria, aunque posee además otros efectos beneficiosos sobre la salud del lactante debido a su elevada proporción de aminoácidos esenciales (triptófano y cisteína). Según diversos estudios parece influir positivamente en la absorción de hierro en el intestino del niño, y en experimentos in vitro, unida al ácido oleico (complejo HAMLET), es efectiva frente a tumores celulares como el papiloma humano. El complejo HAMLET también presenta un claro efecto antimicrobiano frente a Streptococcus pneumoniae, Haemophilus influenzae, cepas enteropatógenas de Escherichia coli y Salmonella thypimurium, sin embargo no se ha demostrado que durante la digestión de la leche materna se forme dicho complejo en el tracto digestivo del lactante. El desarrollo de fórmulas infantiles destinadas a la alimentación del niño durante el primer año de vida ha mejorado considerablemente en las últimas décadas intentando no sólo adecuar la concentración de nutrientes a los requerimientos del lactante, sino también adicionando compuestos bioactivos de diferente naturaleza, como la α-lactoalbúmina, con el objetivo de alcanzar los efectos funcionales que se producen en los niños alimentados con leche materna.

α-Lactalbumin as an ingredient of infant formula. α-lactalbumin is the main whey protein in human milk rising 2,44 g/L in mature milk. It has a key function in the synthesis of lactose from glucose and galactose in the mammary gland although this compound has also other beneficial effects on the infant health due to the high proportion of essential aminoacids (tryptophan and cysteine). It seems also to increase iron absorption in the digestive track, and in in vitro experiments, linked to oleic acid (HAMLET complex), has shown anticarcinogenic effects against cellular tumor such as human papilloma. In addition, this complex has been reported to exhibit antimicrobial properties against Streptococcus pneumoniae,Haemophilus influenzae, enteropathogenic strains of Escherichia coli and Salmonella thypimurium. However, the in vivo synthesis of HAMLET complex during milk digestion has not been proved yet. Infant formula have been improved considerably during the last decades not only adapting nutrient concentrations to infants requirements but also by the addition of new bioactive ingredients such as α-lactalbumin, to have the same functional effect as in breast fed babies.

Role of stabilizers in preparation of a potent sheep pox vaccine

Production of live attenuated sheep pox vaccine sustained the elevated temperatures during freeze-drying, transportation storage and vaccination in unequipped tropical and subtropical zones of the world, is highly recommended. For this reason, eight stabilizer formulas were individually used for preparation of eight sheep pox vaccines, which were lyophilized and then titrated and accordingly four vaccine formulas were eventually selected that should be tested for thermoprotectivity to select the best stabilized vaccine. These selected vaccines were tested for sterility; potency [vaccination and challenge] and safety in susceptible sheep. The collected blood sera were subject to serological examination for estimating the antibody response by ELISA. The results proved transcendence of sheep pox vaccines stabilized with 10% trehalose alone or in combination with 5% lactalbumin hydrolyste in the thermoprotectivey, thereby improvement vaccination efficacy

[Allergic reaction of goats and cows milk proteins]

Systemic anaphylaxis test in mouse showed that the visual effect of injection of cow casein, whey, casein fraction and bovine beta-lactoglobulin was strong. Similar results were found when goat casein, Kappa-casein and beta-casein were injected. The visual effect of injection of beta-lactoglobulin and alpha[s],-casein was low. No response was found when bovine and goat alpha-Lactalbumin was injected in animals. Passive cutaneous anaphylaxis test in mouse showed that the reaction area of cow casein injection was 0.63 cm[2] in comparison with 0.19 cm[2] for goat casein. The reaction of cow beta-lactoglobulin injection was 0.12 cm[2], while no reaction was occurred when goat beta-lactoglobulin, goat and cow alpha-lactalbumin were injected. The percentage of degranulation of mast cells when treated with cow raw milk, casein, whey, beta-lactoglobulin and alpha-lactalbumin were 32.11, 100, 41.80, 90.01 and 12.73% respectively, In comparison with 14.33, 80.19, 34.73, 39.57 and 10.86% respectively for the same proteins in goat milk

[Study of the immunological cross-reactions between goats and cows milk proteins]

Immunoelectrophoresis analysis showed immunological cross reactions between goat and cow milk caseins which belong to beta-casein, however, no such reaction were observed between goat and cow beta-lactoglobulin and alpha-lactalbumin. Systemic anaphylaxis test in guinea pigs showed strong immunological reactions between goat and cow milk proteins, injection of cow milk in animal's vein, which fed on cow milk caused 100% mortality. Same results were also obtained with injection of goat milk. Passive hemagglutination test against goat and cow milk was used to estimate antibody titer in guinea pigs serum, which fed cow's milk. The obtained results showed that the highest titer was found against casein followed by beta-lactoglobulin and alpha-lactalbumin for cow milk proteins, while for goat milk proteins the highest titer was found against casein followed by alpha-lactalbumm and beta-lactoglobulin. The titer of antibodies against goat alpha[s]-casein and Kappa-casein was lower than that for cow milk, the behavior of goat and cow beta-casein was similar for both proteins

Effect of the whey basic protein on bone mineral density / 中华预防医学杂志

<p><b>OBJECTIVE</b>To examine the effects of the whey basic protein on bone metabolism of Sprague-Dawley (SD) rats and healthy mid-aged women.</p><p><b>METHODS</b>Forty-four female SD rats were randomized by weight into four groups of eleven rats each and fed 10 mg x kg BW(-1) x d(-1), 20 mg x kg BW(-1) x d(-1), 30 mg x kg BW(-1) x d(-1) of whey basic protein and control diet was given respectively by intragastrically injection for 90 days. Bone mineral density of femur was measured by dual-energy X-ray absorptiometry in vitro. Sixty-three health women [(37.9 +/- 4.3) years old] were randomly assigned to treatment with placebo, 30 mg whey basic protein per day or 60 mg whey basic protein per day for 24 weeks. The bone mineral density (BMD) of the lumbar vertebrae L2-LA, femoral neck and right calcaneus of each subject were measured by dual-energy X-ray absorptiometry (DXA) at 0 and the 24th week of treatment. Serum bone specific alkaline phosphatase and N-telopeptide (NTX) were measured at 0 and the 14th week.</p><p><b>RESULTS</b>The mean BMD value of the distal end of the femur in 10 mg x kg BW(-1) x d(-1) whey basic protein group was significantly higher than that of the control group at the end of the trail. But after treatment by doses of whey basic protein used in the study, there were no differences between the control group and others groups on bone mineral density in the human trail.</p><p><b>CONCLUSION</b>Whey basic protein should enhance the bone mineral density of the rats' femur and no obvious effect was detected in the human trail.</p>

Single strand confirmation polymorphism (SSCP) detection in exon I of the alfa-lactalbumin gene of Indian Jamunapari milk goats (Capra hircus)

The genetic diversity of Jamunapari goats (Capra hircus) was investigated using an optimized non-radioactive polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) method to detect alfa-lactalbumin polymorphism in a sample of 50 goats. Our data show that PCR-SSCP is an appropriate tool for evaluating genetic variability in Jamunapari goats. Polymorphism was detected in the sample, indicating that Jamunapari goats have high genetic variability at loci, exon I of the a-lactalbumin gene. This result opens interesting prospects for future selection programs and conservation strategies. These a-lactalbumin variants can be sequenced and screened in the population to develop single nucleotide polymorphism (SNP) markers for association studies and marker assisted selection.

A study on the hairy root culture and antitumor alkaloids production of Catharanthus roseus / 中国中药杂志

<p><b>OBJECTIVE</b>To establish transformation system and obtain alkaloids from the hairy root of Catharanthus roseus.</p><p><b>METHOD</b>Hairy roots were obtained by infecting the different explants of C. roseus. Culture conditions of hairy root were optimized.</p><p><b>RESULT</b>The best transformation condition was leaf infected by two-day's pre-culture and two-day's co-culture and additional A(S) (hydroxyacetosyringone) 100 mg x L(-1). The inducing rate of hairy root was up to 86.25%. The best condition of hairy root culture was MS medium with sucrose as carbon material and lactalbumin as nitron material. The analysis result showed that the contents of total alkaloids in hairy roots were higher than explants and calli.</p><p><b>CONCLUSION</b>Hairy root of C. roseus will be useful for the production of active components in C. roseus.</p>

Milk aspiration as a Result of Hypotonic-hyporesponsive Episodes after DTaP and Polio Co-immunization / 대한법의학회지

We analyzed autopsy findings of 5 neonates and infants who suddenly died after DTaP and Polio Coimmunization. Microscopically, there were multifocal small amounts of pinkish amorphous materials within the alveolar spaces with marked proliferation of alveolar macrophages. Immunohistochemical stain using anti-human alpha-lactalbumin shows intense staining to these foreign materials. We concluded that milk aspiration and asphyxia may play an important role in course of death after DTaP and Polio Co-immunization and assumed that hypotonic-hyporesponsive episodes as an adverse events of immunization could be a preceding cause of milk aspiration. Education of parents about the risk of milk aspiration after immunization should be included in guideline.

effect of different stabilizers on maintainig viability of lyophilized pasteurella multocida strains

In the present study, different stabilizers were used. Results indicated that the best viability percentage [70.60 and 68.00], mean death time in mice, microscopical examination and colonial morphology were shown in lyophilized strains stabilized by skimmed milk lactalbumin sucrose peptone medium and Angus medium respectively, followed by 59% and 58% viability in strains stabilized by skimmed milk, lactalbumin, sucrose medium and skimmed milk peptone sucrose medium respectively

Immunocytochemical Study for Lactalbumin in Alveolar Macrophage of Human Milk Aspirated Mouse

PURPOSE: Aspiration of foreign material into the lungs can cause acute or chronic pulmonary diseases. It is difficult to detect small amounts of aspiration due to the lack of safe, sensitive and specific diagnostic tests. Recently, in animal or human studies, it has been reported that immunochemistry for lactalbumin can be used to detect the minimal aspiration. So, the authors' investigation was designed to determine whether human milk phagocytized alveolar macrophages can be detected in human milk aspirated mice. METHODS: Sixty four male mice, 6-8 weeks old and 30-40 gm weighing, were used for this study. About 0.05 mL of human milk or normal saline were given intranasally once per day for 1 day or 3 days. Under anesthesia with ketamine and xylazine, the trachea of each mouse was cannulated with an 18G Jelco needle and then, each mouse's lungs were lavaged three times with 0.5 mL of phosphate buffer solution at 2, 8, 24, and 48 hours after the last milk or normal saline instillation. Cells in bronchoalveolar lavage fluid were stained with Oil Red O and immunocytochemistry for alpha-lactalbumin. RESULTS: Immunocytochemical reactivity for alpha-lactalbumin or lipid-laden alveolar macrophages were not observed in the normal saline aspirated groups. Immunocytochemical reactivity for alpha- lactalbumin were observed in the human milk aspirated groups. They showed a peak at 8 hours and decreased markedly at 24 hours but persisted even at 48 hours after aspiration. Immunocytochemical stain positive alveolar macrophages were noted similarly in number between single and multiple aspiration groups. CONCLUSION: These observations suggested that alveolar macrophages for lactalbumin could be more easily detected on immunocytochemistry than Oil Red O stain, and immunocytochemistry could be used as a sensitive and specific diagnostic test for the detection of human milk aspiration.

Case of a Cystic Hypersecretory Duct Carcinoma of the Breast

Breast cystic hypersecretory duct carcinomas (CHDC), characterized by cystic dilatation of ducts, cysts filled with thyroid colloid-like eosinophilic material, and by micropapillary carcinomas in the epithelium lining the cyst, is a rare histological variant of intraductal carcinomas. CHDC is differentiated from cystic hypersecretory hyperplasia. It's clinical manifestations are similar to those of other intraductal carcinomas. CHDC has a low-grade behavior for many years, but can be invasive, with the potential for metastasis. The importance of CHDC is its differentiation from a variety of other lesions of the breast, both benign and malignant. Various differential diagnoses for CHDC include: fibrocystic changes of the breast with microcyst formation, juvenile papillomatosis, benign mucocele-like lesions, juvenile secretory carcinomas, mucin-producing colloid carcinomas and metastatic follicular thyroid carcinomas. Histochemical staining (PAS, alcian blue and mucicarmine) or immunoperoxidase studies (CEA, alpha-lactalbumin and thyroglobulin) may also helpful in differential diagnoses. We report a case of a cystic hypersecretory duct carcinoma (CHDC) of the breast, and present a review of the literature.

Binding of globular proteins to DNA from surface tension measurement.

Extent of binding (gammap) of globular proteins to calf-thymus DNA have been measured in mole per mole of nucleotide as function of equilibrium protein concentration. We have exploited measurement of the surface tension of the protein solution in the presence and absence of DNA to calculate the binding ration (gammap). Interaction of bovine serum albumin with DNA has been studied at different pH. Interaction of bovine serum albumin with DNA has been studied at different pH, ionic strength and in presence of Ca2+. Interaction of BSA with denatured DNA has also been investigated. Binding isotherms for other globular proteins like beta-lactoglobulin, alpha-lactalbumin and lysozyme have been compared under identical physicochemical condition. It has been noted with considerable interest that globular form of protein is important to some extent in protein-DNA interaction. An attempt has been made to explain the significance of difference in binding ratios of these two biopolymers in aqueous medium for different systems in the light of electrostatic and hydrophobic effects. Values of maximum binding ration (gammap(m)) at saturated level for different systems have been also presented. The Gibb's free energy decrease (-deltaG0) of the binding of proteins to DNA has been compared more precisely for the saturation of binding sites in the DNA with the change of activity of protein in solution from zero to unity in the rational mole fraction scale.

An improved electrophoretic method for the determination of serum milk protein variants in Gyr-Holstein cows

Milk serum proteins such as alpha-lactalbumin (ALA) and beta-lactoglobulin (BLG) present biochemical polymorphism which is under the control of codominant autosomal alleles. In the present report, we propose modifications of traditional electrophoretic techniques such as increasing the running gel concentration from 5 to 10% and the addition of 5 M urea to the stacking gel, which permitted the detection of two variants (A and B) at the ALA and BLG loci. About 8 microliters of milk serum (6 mg/ml protein) and 10 microliters of total fresh milk were applied. Bovine serum albumin (BSA) and immunolactoglobulins (ILG) could also be discriminated. Total fresh milk was as useful as the purified serum milk proteins for the discrimination of ALA and BLG serum milk protein polymorphism by alkaline vertical slab polyacrylamide gel electrophoresis. However, BSA and ILG ran with caseins, which prevented their characterization in this system

Stimulation of the absorption of macromolecules in Giardia lamblia infected mice intestine.

The absorption of 125I-labelled bovine serum albumin, gamma-globulin and alpha-lactalbumin was considerably enhanced in G. lamblia infected Swiss mice intestine compared to uninfected controls. The binding of 125I-proteins to brush border membrane was however, significantly (P less than 0.01) low in infected animals. Kinetic studies with gamma-globulin binding to brush borders revealed a decrease in the number of binding sites in infected animals (1.52) compared to controls (2.86 micrograms/mg protein) with no change in the affinity constant (47.60 micrograms/ml) under these conditions. These findings suggest that G. lamblia infection in mice leads to enhanced macromolecular absorption which seems unrelated to the binding of proteins to epithelial cell surface.