Leukocytes in expressed breast milk of asthmatic mothers.

OBJECTIVE: Infants are born immunologically immature. However, breastfeeding mothers retain an immunological link to their infants. While it is generally accepted that infants are at an immunological advantage when compared with formula-fed infants, the benefit of long-term exclusive breastfeeding by atopic mothers remains controversial. Inconsistency in the conferral of benefit may be due to differences in the immunological constituents passed to the recipient infant. The aim of this investigation was to examine the profile of human milk cells and cytokines from asthmatic compared to non-asthmatic mothers. METHODS: Twenty-five exclusively breastfeeding mothers with a clinical diagnosis of asthma were postpartum age matched in a double-control 2:1 design with 50 non-asthmatic controls. Each mother provided a single milk sample which was assayed for cell differential by flow cytometry, for ex vivo cytokine production in culture and for aqueous phase cytokines. RESULTS: Milks from asthmatic mothers differed from non-asthmatics in that they contained a higher proportion of polymorphonuclear (PMN) cells and lower proportion of lymphocytes, predominantly CD3+/CD4+ T helper cells, reflected by a decrease in the chemokine CCL5 in the milk aqueous phase. More PMN and lymphocytes from asthmatic mothers expressed the adhesion molecule CD11b and lymphocytes the IgE receptor CD23, than those from non-asthmatic mothers. CONCLUSIONS: Changes to human milk leucocyte prevalence, activation state and cytokines due to maternal asthma may result in changes to immunological priming in the infant. Consequently, the protective effect of long-term breastfeeding may be altered in these mother-infant pairs.

Extended boiling of peanut progressively reduces IgE allergenicity while retaining T cell reactivity.

BACKGROUND: Current peanut oral immunotherapy is hampered by frequent adverse events. It has been shown that boiling can reduce peanut allergenicity. Hypoallergenic peanut products have the potential to reduce treatment-related reactions during desensitization. OBJECTIVE: To show that extended boiling (for up to 12 h) can progressively reduce peanut allergenicity while retaining T cell reactivity. METHODS: Raw peanuts were boiled for half, 1, 2, 4 and 12 h in deionized water. After dehydration, boiled and raw peanuts were ground, defatted and soluble proteins extracted in PBS and cooking water (leachate) retained. SDS-PAGE, Western blot, inhibition ELISA, mass spectrometry and skin prick test were used to characterize changes to peanut allergens and human IgE reactivity. T cell responses to raw and boiled peanut extracts were determined by proliferation of CD4+/CD25+/CD134+ T cells in peanut-allergic and non-allergic individuals. RESULTS: Extended boiling progressively reduced peanut allergenicity through a combination of leaching of allergens into cooking water, fragmentation of allergens and denaturation of conformational epitopes. Two-hour boiling led to an eightfold reduction in IgE binding capacity of boiled peanuts as determined by inhibition ELISA, while 12-h boiling led to a 19-fold reduction. Mass spectrometry revealed an increasing number of unique allergen peptides with longer boiling times. Raw, 2- and 12-h boiled peanut extracts were equivalent in their ability to stimulate T cell activation and proliferation. CONCLUSION AND CLINICAL RELEVANCE: Progressive reduction in peanut allergenicity with extended boiling does not affect T cell reactivity. Boiled peanuts may be a candidate for oral immunotherapy.

Assessment and management of pain in children and adolescents with bleeding disorders: a cross-sectional study from three haemophilia centres.

INTRODUCTION: Pain is a major clinical problem in patients with bleeding disorders. This study aimed to determine the level of pain and how it is managed in children and adolescents with bleeding disorders. METHODS: This cross-sectional study was performed in three haemophilia centres (one in Shiraz and two in London).The data were collected using questions about pain management and Wong-Baker Faces Pain Rating Scale at routine clinical review as well as attendance for bleed treatment in summer 2014. RESULTS: This study indicated no difference among the three haemophilia centres regarding having pain, however, a significant difference was found among them concerning the mean score of pain intensity. Among the 154 subjects, 20.8% had pain on the study day, most reporting moderate levels of pain. The study participant's most frequently described their pain as aching, dull, throbbing and stabbing. Moreover, 84.38% of pain was experienced in joints and the most common painful joints were knees, ankles, elbows, hands and hips. The most common pain relief strategies included factor administration, immobilization and rest, ice packs and analgesia. Pain was significantly associated with disease severity and age. CONCLUSION: As the intensity of pain in on-demand patients was highest, using prophylaxis treatment is suggested. Moreover, adolescent patients reported more pain; giving more self-care information to them and their parents is recommended. Since little evidence was published for pain assessment and management in children and adolescents with bleeding disorders, more research is recommended.

Silos and rigid processes: Barriers to the successful implementation of the Older prisoner Health and Social Care Assessment and Plan.

Older adults are the fastest growing sub-group in prisons. They have complex health, social care and custodial needs and often the support they receive is sub-optimal. The Older prisoner Health and Social Care Assessment and Plan (OHSCAP) aimed to better meet these inter-related needs. As part of a wider study, a randomised controlled trial was conducted to evaluate the OHSCAPs effectiveness in meeting older prisoners' health, social care and custodial needs in comparison to treatment as usual. This article describes the nested qualitative study which aimed to explore the barriers and facilitators to the effective implementation of the OHSCAP. Semi-structured interviews were conducted with older adults (n = 14) and staff members t (n = 12). Data was analysed using the framework method. Three overarching key themes were identified. These were: (1) balancing care and custodial requirements; (2) prison, health and social care silos; and (3) rigid prison processes. Prison is an important opportunity to engage residents and improve public health. Cultural and strategic change is required for health, social care and custodial interventions, such as the OHSCAP, to be successfully implemented into prison settings.

Services for children with developmental co-ordination disorder: the experiences of parents.

BACKGROUND: Parents provide valuable information on their experiences of engaging with therapy services for their children, which can inform the future development of these services. The aim of this study was to explore the views and experiences of parents who had accessed therapy services for their child with developmental co-ordination disorder (DCD). METHODS: Seven focus groups were conducted incorporating 52 parents who had a child diagnosed with, or fitting the diagnostic criteria for DCD. Focus groups were audiotaped, transcribed and analysed thematically. FINDINGS: Parents reported struggling to gain access to therapy services. When they gained access, they found the services beneficial for their child but continued to experience difficulties regarding the quality of service delivery. CONCLUSIONS/IMPLICATIONS: The study suggests that parents thought some health-care professionals lacked knowledge and understanding of DCD, which they believed impacted upon early recognition and access to services. They perceived that therapy at an early age was vital for children's development, and indicated that a clearer path for accessing these services was necessary in addition to improved service quality. They called for an increase in awareness of DCD by all therapy service professionals to aid early recognition and improved treatment.

Knob-independent cytoadherence of Plasmodium falciparum to the leukocyte differentiation antigen CD36.

The survival of Plasmodium falciparum-infected erythrocytes is enhanced by the sequestration of mature trophozoites and schizonts from the peripheral circulation. Cytoadherence of infected erythrocytes in vivo is associated with the presence of knobs on the erythrocyte surface, but we and others have shown recently that cytoadherence to C32 melanoma cells may occur in vitro in the absence of knobs. We show here that a knobless clone of P. falciparum adheres to the leukocyte differentiation antigen, CD36, suggesting that binding to CD36 is independent of the presence of knobs on the surface of the infected erythrocyte. This clone showed little cytoadherence to immobilized thrombospondin or to endothelial cells expressing the intercellular adhesion molecule 1. Furthermore, an Mr approximately 300-kD trypsin-sensitive protein doublet was immunoprecipitated from knobless trophozoite-infected erythrocytes. Finding a P. falciparum erythrocyte membrane protein 1 (PfEMP1)-like molecule on these infected erythrocytes is consistent with a role for PfEMP1 in cytoadherence to CD36 and C32 melanoma cells.

Specificity for aminoacylation of an RNA helix: an unpaired, exocyclic amino group in the minor groove.

An acceptor stem G3.U70 base pair is a major determinant of the identity of an alanine transfer RNA. Hairpin helices and RNA duplexes consisting of complementary single strands are aminoacylated with alanine if they contain G3.U70. Chemical synthesis of RNA duplexes enabled the introduction of base analogs that tested the role of specific functional groups in the major and minor grooves of the RNA helix. The results of these experiments indicate that an unpaired guanine 2-amino group at a specific position in the minor groove of an RNA helix marks a molecule for aminoacylation with alanine.

Differential recognition of a protective filarial antigen by antibodies from humans with bancroftian filariasis.

The objectives of this study were to identify filarial antigens which induce enhanced clearance of circulating microfilariae and to establish if human antibody reactivity with these molecules correlates with the apparent parasite burdens of residents of an endemic area of Bancroftian filariasis. Mice immunized with an extract of Brugia malayi microfilariae develop IgG antibodies to four major filarial antigens with an apparent molecular weight (Mr) of approximately 112,000, 60,000, 45,000, and 25,000. Animals immunized with gel slices containing the approximately 25,000-Mr antigen are resistant to intravenous challenge with live microfilariae (78-98% reduction in parasitemia vs. controls, P less than 0.01). A group of 22 amicrofilaremic humans had a significantly higher (P less than 0.025) mean antibody titer to the Mr 25,000-Mr antigen (1: 424) than 16 microfilaremic individuals (1:95). There were no significant differences between the two groups in antibody titers to filarial antigens of Mr approximately 112,000, 60,000, and 45,000 Mr. These data suggest that a high degree of reactivity to the 25,000-Mr antigen in humans with lymphatic filariasis correlates with a parasitologic status that is least conducive to transmission of infection.

Efficient aminoacylation of tRNA(Lys,3) by human lysyl-tRNA synthetase is dependent on covalent continuity between the acceptor stem and the anticodon domain.

In this work, we probe the role of the anticodon in tRNA recognition by human lysyl-tRNA synthetase (hLysRS). Large decreases in aminoacylation efficiency are observed upon mutagenesis of anticodon positions U35 and U36 of human tRNA(Lys,3). A minihelix derived from the acceptor-TPsiC stem-loop domain of human tRNA(Lys,3)was not specifically aminoacylated by the human enzyme. The presence of an anticodon-derived stem-loop failed to stimulate aminoacylation of the minihelix. Thus, covalent continuity between the acceptor stem and anticodon domains appears to be an important requirement for efficient charging by hLysRS. To further examine the mechanism of communication between the critical anticodon recognition elements and the catalytic site, a two piece semi-synthetic tRNA(Lys, 3)construct was used. The wild-type semi-synthetic tRNA contained a break in the phosphodiester backbone in the D loop and was an efficient substrate for hLysRS. In contrast, a truncated variant that lacked nucleotides 8-17 in the D stem-loop displayedseverely reduced catalytic efficiency. The elimination of key tRNA tertiary structural elements has little effect on anticodon-dependent substrate binding but severely impacts formation of the proper transition state for catalysis. Taken together, our studies provide new insights into human tRNA structural requirements for effective transmission of the anticodon recognition signal to the distal acceptor stem domain.