International consensus guidelines for the diagnosis and management of food protein-induced enterocolitis syndrome: Executive summary-Workgroup Report of the Adverse Reactions to Foods Committee, American Academy of Allergy, Asthma & Immunology.

Food protein-induced enterocolitis (FPIES) is a non-IgE cell- mediated food allergy that can be severe and lead to shock. Despite the potential seriousness of reactions, awareness of FPIES is low; high-quality studies providing insight into the pathophysiology, diagnosis, and management are lacking; and clinical outcomes are poorly established. This consensus document is the result of work done by an international workgroup convened through the Adverse Reactions to Foods Committee of the American Academy of Allergy, Asthma & Immunology and the International FPIES Association advocacy group. These are the first international evidence-based guidelines to improve the diagnosis and management of patients with FPIES. Research on prevalence, pathophysiology, diagnostic markers, and future treatments is necessary to improve the care of patients with FPIES. These guidelines will be updated periodically as more evidence becomes available.

Alternative glycosylation of the insulin receptor prevents oligomerization and acquisition of insulin-dependent tyrosine kinase activity.

Glucose deprivation leads to the synthesis of an aberrantly glycosylated ('alternative') and inefficiently processed form of the insulin proreceptor in 3T3-L1 adipocytes. To further explore the effect of aberrant (rather than absent) N-linked glycosylation of the insulin receptor, we examined the relationship of processing to function. Our studies show that the alternative form of the proreceptor does not oligomerize nor does it acquire the ability to undergo insulin-sensitive autophosphorylation. This along with an interaction with the glucose-regulated stress protein GRP78/BiP implies inappropriate folding/dimerization and retention in the ER. Glucose refeeding causes the post-translational modification of the alternative form of the proreceptor to a novel 'intermediate' form which is independent of new protein synthesis. As little as 100 microM glucose (or mannose) can induce this modification. In vitro digestion of the alternative and intermediate proreceptors with SPC1/furin shows that both the alpha- and beta-subunit domains are glycosylated, albeit aberrantly. This implies that the aberrantly glycosylated proreceptor could serve as a substrate for SPC1 in a physiological setting if the receptor was able to interact with the enzyme in the appropriate compartment (i.e., the trans-Golgi network). Based on inhibitor studies, however, both the alternative and intermediate forms of the proreceptor appear to be primarily targeted to the proteasome for degradation.

Prospective follow-up oral food challenge in food protein-induced enterocolitis syndrome.

OBJECTIVES: To determine tolerance rates to cow's milk and soy and to suggest guidelines for follow-up oral food challenges (FU-OFCs) in infantile food protein-induced enterocolitis syndrome (FPIES). METHODS: The authors analysed the data of 23 patients with infantile FPIES who underwent two or more FU-OFCs and were followed up until over 2 years of age. The first FU-OFCs were performed at 6 months of age, and patients were randomly allocated to cow's milk (n = 11) or soy (n = 12) challenge starting groups. Second and third FU-OFCs were performed at 2-month intervals in a crossed and switched-over manner. RESULTS: Tolerance rates to cow's milk and soy were 27.3% and 75.0% at 6 months of age, 41.7% and 90.9% at 8 months and 63.6% and 91.7% at 10 months, respectively. Patients outgrew cow's milk and soy intolerance at age 20 and 14 months. CONCLUSIONS: In infantile FPIES, the first FU-OFC should be performed with soy at 6-8 months of age and cow's milk FU-OFC should be conducted at over 12 months of age. Infants with FPIES were observed to outgrow food sensitivities during the first 2 years of life.

Effect of alternative glycosylation on insulin receptor processing.

The mature insulin receptor is a cell surface heterotetrameric glycoprotein composed of two alpha- and two beta-subunits. In 3T3-L1 adipocytes as in other cell types, the receptor is synthesized as a single polypeptide consisting of uncleaved alpha- and beta-subunits, migrating as a 190-kDa glycoprotein. To examine the importance of N-linked glycosylation on insulin receptor processing, we have used glucose deprivation as a tool to alter protein glycosylation. Western blot analysis shows that glucose deprivation led to a time-dependent accumulation of an alternative proreceptor of 170 kDa in a subcellular fraction consistent with endoplasmic reticulum localization. Co-precipitation assays provide evidence that the alternative proreceptor bound GRP78, an endoplasmic reticulum molecular chaperone. N-Glycosidase F treatment shows that the alternative proreceptor contained N-linked oligosaccharides. Yet, endoglycosidase H insensitivity indicates an aberrant oligosaccharide structure. Using pulse-chase methodology, we show that the synthetic rate was similar between the normal and alternative proreceptor. However, the normal proreceptor was processed into alpha- and beta-subunits (t((1)/(2)) = 1.3 +/- 0.6 h), while the alternative proreceptor was degraded (t((1)/(2)) = 5.1 +/- 0.6 h). Upon refeeding cells that were initially deprived of glucose, the alternative proreceptor was processed to a higher molecular weight form and gained sensitivity to endoglycosidase H. This "intermediate" form of the proreceptor was also degraded, although a small fraction escaped degradation, resulting in cleavage to the alpha- and beta-subunits. These data provide evidence for the first time that glucose deprivation leads to the accumulation of an alternative proreceptor, which can be post-translationally glycosylated with the readdition of glucose inducing both accelerated degradation and maturation.

Glucose deprivation does not affect GLUT1 targeting in 3T3-L1 adipocytes.

We have previously demonstrated that glucose deprivation alters the glycosylation of the GLUT1 glucose transporter in 3T3-L1 adipocytes. Many aberrantly glycosylated proteins are retained in the endoplasmic reticulum by interaction with chaperones. Herein, we use three independent procedures to show that GLUT1 is targeted to the plasma membrane, despite alterations in glycosylation. While earlier experiments revealed that plasma membrane targeting of aglyco GLUT 1 transporter was significantly reduced, our data show for the first time that altered glycosylation provides sufficient information to drive appropriate trafficking.

Mass transfer from gas bubbles to impinging flow of biological fluids with chemical reaction.

The rates of mass transfer from a gas bubble to an impinging flow of a biological fluid such as whole blood and plasma are investigated analytically and experimentally. Gases commonly found dissolved in body fluids are included. Consideration is given to the effects of the chemical reaction between the dissolved gas and the liquid on the rate of mass transfer. Through the application of boundary layer theory the over-all transfer is found to be Sh/(Re)(1/2) = 0.845 Sc(1/3) in the absence of chemical reaction, and Sh/(Re) (1/2) = F' (0) in the presence of chemical reaction, where Sh, Re, and Sc are the Sherwood, Reynolds, and Schmidt numbers, respectively, and F' (0) is a function of Sc and the dimensionless reaction rate constant. Analytical results are also obtained for the bubble lifetime and the bubble radius-time history. These results, which are not incompatible with experimental results, can be applied to predict the dissolution of the entrapped gas emboli in the circulatory system of the human body.

Deposition of eosinophil-granule major basic protein and expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in the mucosa of the small intestine in infants with cow's milk-sensitive enteropathy.

BACKGROUND: Cow's milk-sensitive enteropathy (CMSE) is an important cause of chronic diarrhea and failure to thrive in infancy. The immunopathology of the mucosal lesion associated with CMSE has not yet been described. OBJECTIVES: This study investigated the eosinophil activation and the role of adhesion molecules in the pathogenesis of intestinal mucosal damage associated with CMSE. METHODS: Twenty-one patients with chronic diarrhea and abnormal mucosa on duodenal biopsy specimens were included. The patients had negative responses to skin prick tests and RASTs with milk. Fourteen patients were diagnosed with CMSE by milk challenge test and were designated as the CMSE group. Seven patients with no milk intolerance were defined as the non-CMSE group. Four infants with frequent vomiting and no mucosal abnormalities were also studied as the control group. Immunohistochemical stains for eosinophil major basic protein (MBP), vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 on endoscopic duodenal biopsy specimens were performed. RESULTS: The degree of eosinophil degranulation, as evidenced by localization of extracellular MBP, was significantly greater in the CMSE group compared with the non-CMSE and control groups (P <.05). Expression of VCAM-1 on mononuclear cells was higher in the CMSE group compared with the non-CMSE and control groups (P <.05). The severity of villous atrophy was positively correlated with the deposition of MBP (r = 0.79, P <.001). CONCLUSION: These results strongly suggest eosinophils and VCAM-1 are implicated in the pathogenesis of mucosal damage associated with CMSE.