A rapid objective method for measuring the yeast opsonisation activity of serum.

A simple, objective semi-quantitative assay for yeast opsonisation by normal polymorphs has been developed. This depends on electronically counting the number of free unphagocytosed yeast particles on a Coulter counter. The method has been compared with the widely used microscope technique and shows excellent correlation. The sera of 112 unselected school-children gave a distribution curve consistent with 2 peaks; 7 gave values less than 2 S.D.s below the mean confirming the high incidence of this immunodeficiency.

Real time breath and headspace analysis of flavour volatiles.

An overview is presented of the principle, scope and major applications to date of the use of atmospheric pressure chemical ionisation mass spectrometry (APCI-MS) for monitoring the kinetics of release of flavour volatiles in real time, principally from breath during eating. The technique is rapid, quantitative, sensitive to the ppb level and can be used to monitor the vast majority of flavour volatiles. Advances made during the last 5 years in our understanding of factors affecting flavour release, particularly when conducted simultaneously with sensory evaluation, are contributing increasingly to more efficient product development in the food and flavour industry and to the design of flavour systems with desired dynamic flavour characteristics. Real time APCI-MS headspace data may be used to validate mathematical modelling of flavour release. It is proposed that these advances may be applied with similar benefits in the pharmaceutical industry, particularly in the improvement of the flavour acceptability of orally administered drugs.

Defective opsonization. A common immunity deficiency.

Serum opsonization of yeasts for phagocytosis by normal polymorphonuclear leucocytes was defective in 11 of 43 children with unexplained frequent infections. The children had a range of infections, largely bacterial, and only 3 had diarrhoea and rash in infancy. A similar defect in at least 6 of the 9 mothers of these children (of either sex), with normal function in the fathers, suggests that the defect was primary and was transmitted by an unusual form of dominant inheritance. Four of 72 healthy adults and 1 of 11 children with unrelated disease showed similar defective function, but the incidence of the defect in the patients with frequent infection was significantly greater than this. The defective function can be corrected, in vitro and in vivo, by normal plasma at concentrations too low to be effective alone. This suggests that there is a defective factor rather than an inhibitor, and that different factors are limiting in normal and in defective plasma. Sera from affected members of the same family do not correct each other, but defective sera from different families usually do.

A defect of the alternative pathway of complement.

The sera of patients defective at opsonization for yeast phagocytosis fix haemolytic complement poorly following incubation with the alternative pathway complement activators, inulin and bacterial endotoxin. This suggests that the defect lies early in the alternative pathway of complement.

Clinicopathological findings in patients with primary and secondary defects of neutrophil mobility.

Twenty-eight patients with defective neutrophil mobility were classified, largely on results of parent values, into primary (17)--mainly Shwachman's syndrome--and probable secondary (11). They presented with frequent infections and/or allergy and these symptoms were essentially similar in both groups. Neutropenia was virtually confined. to the patients with Shwachman's syndrome. Diagnoses associated with secondary neutrophil mobility defects included hydrocephalus with shunt, cystic fibrosis, and immunoglobulin deficiency. The neutrophil mobility of nearly nearly all the patients studied increased when levamisole was added in vitro, but there was no clear evidence of clinical benefit when patients were given the drug.

Phagocytosis and killing of bacteria and yeast by human milk cells after opsonisation in aqueous phase of milk.

Macrophages and neutrophils from human milk phagocytose and kill Staphylococcus aureus and Escherichia coli in vitro after opsonisation by the aqueous phase of milk as effectively as blood leucocytes in serum. They also phagocytose Candida albicans. The overgrowth of E coli resulting from the addition of iron to cultures of the organism in the aqueous phase of milk is not influenced by the presence of cells. We conclude that the phagocytosis and killing of bacteria by milk cells may contribute to the lower incidence of infection among breast-fed than artificially fed babies.

A polymorph bactericidal defect and a lupus-like syndrome.

We describe a child with primary defect of polymorph bacterial killing associated with systemic lungs erythematosus. We suggest that her autoimmune disease results from chronic bacterial antigen stimulation and propose a hypothetical model linking immunodeficiency with autoimmunity.

Neutrophil mobility during anaesthesia in children. A trial of ascorbate premedication.

In 20 healthy children undergoing elective surgery, mobility of neutrophils, both unstimulated and stimulated by endotoxin, was studied using a millipore filter system with microscopic determination of leading front migration. Paired samples were incubated with 10(-2) mol l-1 calcium ascorbate and ten children also received 10 mg kg-1 ascorbic acid before premedication. Stimulation of mobility was reduced after the opioid premedication (P less than 0.05) in the ascorbate group only, but not significantly during anaesthesia and surgery. A few individuals showed persisting abnormally low values. No effect of ascorbate in vivo or in vitro was demonstrated. There were no infections.

Phorbol myristate acetate stimulated NBT test: a simple method suitable for antenatal diagnosis of chronic granulomatous disease.

When endotoxin was compared with phorbol myristate acetate (PMA) for stimulation of phagocytes in the nitroblue tetrazolium (NBT) test, both methods discriminated between affected patients with X-linked chronic granulomatous disease (CGD) and controls, but only the PMA NBT test distinguished female carriers of CGD. Endotoxin provided no stimulation of normal fetal blood whereas PMA was an effective stimulator. Our results indicate the superiority of the PMA NBT test for diagnosis of patients and carriers of CGD and should allow accurate antenatal diagnosis of the disease.

An inherited defect of neutrophil mobility in Shwachman syndrome.

Selected immunologic functions were assessed in 14 patients with the Shwachman syndrome. Nine patients were neutropenic and four had low levels of IgA or of IgM. Neutrophil mobility was significantly defective in the group of patients as a whole (in 12 it was below the lower limit of normal) and in their parents. No other consistent abnormality in immunity was found. These results suggest that the defective neutrophil mobility is a feature of Shwachman syndrome which may contribute to the vulnerability of these patients to frequent infections. The defect appears to be a primary genetic one, inherited as an autosomal recessive characteristic consistent with the assumed inheritance of Shwachman syndrome.

Maternal smoking and cord blood immunity function.

Thymidine uptake in PHA-stimulated culture of cord blood cells from smoking mothers is greater than that from non-smoking mothers. There was no such difference when separated lymphocytes were studied; this suggests that smoking suppresses a suppressor cell. No difference was detected in haemoglobin, immunoglobulins, transferrin, cell counts, E-rosette counts and leucocyte mobility.

Familial opsonization defect associated with fatal infantile dermatitis, infections, and histiocytosis.

Members of four generations of a family had a defect of serum opsonization for yeast phagocytosis consistent with dominant inheritance. 2 were healthy, one had chronic osteomyelitis, and the fourth developed a fatal illness in infancy characterized by exfoliative dermatitis, diarrhoea, multiple bacterial infections, and failure to thrive, which resembled the two prevously reported cases with this opsonization defect. At necropsy the infant also had lymphoid depletion, which was possibly secondary, and massive histiocytic infiltration.

Hypogammaglobulinaemia and malakoplakia: response to bethanechol.

A second patient is described with the syndrome of hypogammaglobulinaemia and malakoplakia; he too responded dramatically to bethanechol treatment and remains well on it.

Delayed separation of the umbilical cord, widespread infections, and defective neutrophil mobility.

In six infants, from two families. the umbilical cords were still attached at 3 weeks of age. Five of these developed severe local and disseminated infections from which four died. Two of these children were tested, and both, including the survivor, had defective neutrophil mobility; in the survivor this was improved in vitro and in vivo by ascorbic acid. It is suggested that a primary genetic defect of a contractile protein could explain the association. The sixth child, with delayed cord separation but normal neutrophil mobility and no excess of infections, who has survived without special treatments, also has mastocytosis, apparently inherited independently.

Defective yeast opsonization and C2 deficiency in atopic patients.

Twenty-seven per cent of atopic patients initially presenting with infantile eczema or hay fever were defective for yeast opsonization and 18% had low levels of C2; these deficiencies were mutually exclusive, suggesting that they are primary. Both defects were associated with each of four different atopic syndromes, some of which were related to certain HLA haplotypes.

Yeast opsonisation in children with chronic diarrhoeal states.

Four patients with defective yeast opsonisation and protracted diarrhoea are reported. Plasma infusions improved the opsonising function in all 4 and the diarrhoea in 3. This immunological abnormality was assessed in 100 sequential patients with chronic diarrhoea associated with various gastrointestinal disorders; 52 with protracted diarrhoea and failure to thrive of undetermined cause, 26 with 'toddler diarrhoea', 8 with coeliac disease, 5 with chronic inflammatory bowel disease, and 9 with miscellaneous disorders. 23% of the patients with protracted diarrhoea of undetermined cause had defective opsonisation, a greater proportion (P less than 0.05) than that in 'toddler diarrhoea' or the remaining patients, in whom the frequency (4%) was similar to that (5%) in healthy populations. We suggest that yeast opsonisation be tested in children with protracted diarrhoea, as plasma infusions can be an effective form of treatment.

Monosomy 7 in childhood: a myeloproliferative disorder.

Between 1976 and 1979 a myeloproliferative disease associated with cells monosomic for chromosome number 7 in the bone marrow was seen in six boys aged 5 1/2 months to 8 years (median 10 months). Presenting features included hepatosplenomegaly (5/6), respiratory infections (4/6), pallor (2/6) and skin infections (1/6). Haematological features included a leucoerythroblastic anaemia with leucocytosis and thrombocytopenia, and a hyperplastic marrow with a slight excess of blasts. Fetal haemoglobin was normal in four patients and mildly raised in the other two. Neutrophil function tests showed defective chemotaxis with reduced killing, despite a normal NBT test. Cytogenetic analysis of the marrow showed a preponderance of cells with monosomy 7; the blood lymphocytes were cytogenetically normal. In three patients the disease progressed to acute myeloid leukaemia (AML) after 3 weeks to 23 months; the only patient who remitted did so in response to 6-mercaptopurine and prednisolone, but relapsed 16 months later. A fourth child developed massive splenomegaly which initially responded to 6-mercaptopurine and prednisolone, but progressed to myelofibrosis 11 months later. A fifth child died from anaemia and respiratory infection without progression to leukaemia and the sixth patient has not yet developed leukaemia. Monosomy 7 is the diagnostic criterion of one of the more common myeloproliferative states in childhood and carries a high risk of progression to AML. The acute phase is usually resistant to chemotherapy, but even in responsive cases treatment does not result in elimination of the abnormal clone. Allogeneic bone marrow transplantation should be considered in cases with a suitable donor.

Leukaemia in children and their grandparents: studies of immune function in six families.

Seven of 500 children with acute leukaemia seen over a 15-year period were known to have a close relative with leukaemia or lymphoma. In each case the affected relative was a grandparent of the child, six of the seven being paternal grandparents. Investigation of thses six families showed that the fathers, who had two affected first-degree relatives, had lower lymphocyte counts and higher serum IgA concentrations than paired controls. Atopy, repeated infections and rheumatic disease were common amongst the parents and their sibs. The findings suggest a possible immunodeficiency basis for leukaemia in these families and perhaps also for acute lymphoblastic leukaemia of childhood in general. In the only family in which three generations, including both leukaemic patients, were available for HL-A typing, the affected grandson had not inherited either of his affected grandmother's haplotypes.