Production of pyroglutamic acid by thermophilic lactic acid bacteria in hard-cooked mini-cheeses.

Pyroglutamic acid is present in high amounts (0.5g/ 100g) in many cheese varieties-and particularly in extensively ripened Italian cheeses such as Grana Padano and Parmigiano Reggiano. An in vivo model system for cooked mini-cheese production and ripening acceleration was set up to demonstrate the ability of thermophilic lactic acid bacteria, used as a starter, to produce pyroglutamic acid (pGlu). In mini-cheeses stored at 38 and 30 degrees C for up to 45 d, all starters tested produced different amounts of pGlu. In descending order of pGlu production, the bacteria analyzed were: Lactobacillus helveticus, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and Lactobacillus delbrueckii subsp. lactis. Evidence for the presence of glutamine to pGlu cyclase activity in lactic acid bacteria was provided. Cell lysates obtained from cultures of L. helveticus, L. delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, and S. thermophilus showed the ability to cyclize glutamine to pGlu, resulting in processing yields from 1.4 to 30.3%, depending on the subspecies. Formation of pGlu from free glutamine appeared to be similar to that observed using a glutamine-glutamine dipeptide substrate. Under the experimental conditions applied, pGlu aminopeptidase activity was only detected in L. helveticus. Thus, pGlu formation in long-ripened cooked cheese may depend on the activity of thermophilic lactic acid bacteria.

Pyroglutamic acid in cheese: presence, origin, and correlation with ripening time of Grana Padano cheese.

Pyroglutamic acid is present in many cheese varieties and particularly in high amounts (0.5 g/100 g of cheese) in extensively ripened Italian cheeses (Grana Padano and Parmigiano Reggiano) that are produced with thermophilic lactic acid bacteria as starters. The mechanism of pyroglutamic acid formation in cheese seems to be mostly enzymatic, as demonstrated by the presence of only L-pyroglutamic acid enantiomer. Thermophilic lactobacilli are involved in pyroglutamic acid production, as suggested by the low pyroglutamic acid content found in Bagos, a ripened Italian mountain cheese produced without addition of starter. Because milk pasteurization did not influence the pyroglutamic acid content in the ripened Grana Padano cheese, the formation of pyroglutamic acid mainly depends on the whey starter microflora rather than that of raw milk. Pyroglutamic acid concentration is linearly correlated (R2 = 0.94) with the age of Grana Padano cheese.

Hypersensitivity reactions from inhalation of milk proteins.

We describe the case of a young man with IGE-mediated hypersensitivity to milk, casein, and lactoglobulin, who went into respiratory crisis every time he milked his sheep.

Ultrastructural, immunologic and clinical follow-up of five patients with HCL treated with interferon (IFN) for more than three years.

BACKGROUND: Treatment results in HCl have been improved by the use of alpha-IFN, which is now the standard first-line therapy for this disease, but the mechanism of IFN action is still unclear. It is known, however, that IFN is able to induce hematologic, immunological and phenotype membrane changes which parallel the patients' (pts) clinical response. The aim of our study was to correlate the clinical response to IFN treatment with ultrastructural and phenotype membrane changes in hairy cells (HCs), in order to elucidate the mechanism of IFN action at the cell level. METHODS: We assessed the phenotype membrane and ultrastructural changes induced in HCs by long-term alpha-IFN treatment in five pts with HCL; membrane-bound Il 2-R on PHA-stimulated PBL, the release of IL 2-R by PHA-stimulated PBL and the serum levels of s-IL 2-R were also determined in one pt. RESULTS: The surface immunological phenotype, mainly the HCL-related surface antigen CD25, changed after IFN treatment, dropping from abnormally high to normal values. Furthermore, IFN treatment induced ultrastructural changes in HCs, consisting mainly of a sharp reduction in, up to the almost complete disappearance of, the hairy projections: very few, if any, short, thick villi persisted. The ultrastructural changes in HCs paralleled clinical and hematologic response to IFN treatment in such a way that IFN alone may be considered the cause of these changes. As far as detection of the membrane-bound IL 2-R p55 chain on PHA-stimulated PBL is concerned, the expression of p55 is very high on the membrane of HCs; a high level of serum s-IL 2-R was also found in the HCL pt studied before IFN treatment, whereas the release of IL 2-R by PHA-stimulated PBL was higher than normal, but not significantly. Two of our pts, who did not respond or responded very poorly clinically to IFN treatment, should probably be considered cases of HCL "variants". CONCLUSIONS: The phenotype membrane and the ultrastructural changes in HCs very closely paralleled the patients' clinical responses to IFN, suggesting that both the immunologic and the morphologic changes induced in HCs by in vivo IFN treatment are a direct counterpart of its biologic effect.

Treatment of chronic idiopathic urticaria with topical preparations: controlled study of oxatomide gel versus dechlorpheniramine cream.

A controlled and completely randomized study was carried out with the aim of assessing the efficacy and safety of oxatomide gel in comparison with another preparation for topical use, dechlorpheniramine. Twenty-seven patients (sixteen F, eleven M) aged between 21 and 72 years (mean age 39) suffering from chronic idiopathic urticaria were treated for 15 days with oxatomide gel at 5% or dechlorpheniramine cream at 1%; 15 days of follow-up without therapy were then observed. Both the treatments allowed significant control of cutaneous symptoms. In particular, in the group treated with oxatomide there was a more marked reduction in itching and in the number of weals (p less than 0.01 between times), and in the dechlorpheniramine group in the severity of erythema (p less than 0.01 between times). During the follow-up period, a distinct flare-up of symptoms was observed only in the dechlorpheniramine group. Acceptability and safety, both clinical and biological, were good for both products.

Prolonged treatment with acrivastine for seasonal allergic rhinitis.

In a double-blind, placebo-controlled multicentre study, the antihistamine acrivastine, was used over prolonged periods for the treatment of seasonal allergic rhinitis. After the initial treatment period of 10 days, 8 mg acrivastine three times daily was significantly superior to placebo in controlling the symptoms of sneezing, itchy nose, running nose, watery eyes, itchy eyes and itchy throat. The benefit from acrivastine was also apparent in the second (14 days) and third (28 days) treatment periods, although the difference no longer reached statistical significance. This was probably due to the large proportion of non-responders in the placebo group who withdrew from the study owing to lack of efficacy. The investigators rated symptom control with acrivastine to be 'good' in comparison to 'poor' control with placebo treatment (P = 0.01) for all three periods. There were no significant differences between acrivastine and placebo in the incidence of adverse experiences at the end of each treatment period. Acrivastine is effective and well tolerated over prolonged periods (up to 52 days) for the treatment of seasonal allergic rhinitis.

Immunologic assessment determined by response to IL-2 and immunophenotyping of tumor-infiltrating lymphocytes, of invaded and non invaded lymph nodes and of peripheral blood lymphocytes from twenty-one patients with primary laryngeal cancer.

The aim of the study was to identify the lymphocyte sets and/or subsets possibly involved in the response to malignant cells. For this purpose we have investigated both the cells at the tumor site, i.e. tumor-infiltrating lymphocytes (TIL), and the cells present in the draining lymph nodes, either invaded or non invaded, as well as the peripheral blood lymphocytes from twenty-one patients with primary laryngeal epidermoid carcinoma. The functional assay was carried out by the proliferative response to mitogens, to Interleukin 2 and to their association, the surface immunophenotyping was performed with a large panel of monoclonal antibodies. TIL are the most responsive cells to mitogens, while the responsiveness of TIL and of LN cells to IL-2 was in the same range. PHA-activated TIL are the most responsive cells to IL-2. Our data indicate that TIL do show in vitro, and probably also in vivo, "activation" with elevated responsiveness to IL-2. The surface phenotype showed a strikingly increased proportion of T8+ cells in TIL as compared to T8+ cells in all types of LN, thus confirming within TIL variable, but high, proportions of clones which display cytolytic activity, possibly induced by IL-2. Our data seem to support the perspective for a therapeutic approach in vivo with IL-2, which via its influence on TIL, may act on tumor cells.

Peripheral blood lymphocyte response to recombinant and non-recombinant interleukin 2 in previously treated patients with Hodgkin's disease, long-time off-therapy.

13 patients with Hodgkin's disease (HD) previously treated, 9 of whom were long-time (more than 2 yr) off-therapy, were studied for peripheral blood lymphocyte response to interleukin 2 and for lymphocyte subpopulations by means of in vitro cultures and monoclonal antibodies. The aim of the study was to ascertain the role played by interleukin 2 in the impaired cell-mediated immunity of HD patients. The results show a response of peripheral blood mononuclear cells of HD patients to either the T cell-specific polyclonal mitogens PHA and Con A or to the T cell-dependent, although B cell-specific, PWM, most significantly decreased compared to the normal response. As far as the interleukin 2 involvement in HD is concerned, our study suggests: an impaired endogenous interleukin 2 production by T lymphocytes, a most probable deficiency of the interleukin 2 receptor (Tac) expression and 3) a decrease of the number and/or of the function of NK cells no longer responsive in vitro to interleukin 2. The phenotypic analysis of peripheral blood mononuclear cells showed a slight decrease of total T cells (T3+), of the helper/inducer subset (T4+) and of the T4+/T8+ cells ratio. Our data seem to support the rationale for a therapeutical approach with interleukin 2 in controlled clinical trials also in HD patients, according to the experiments in progress in solid tumor patients.

Cryoglobulinemias and connective tissue diseases.

Cryoglobulinemias in connective tissue diseases (CTD) represent, according to various authors, 12-30% of all cryoglobulinemia cases. Among CTD, systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and Sjögren's syndrome are the diseases most frequently involved in the presence of cryoglobulins (8-48%). The cryoglobulinemias are mostly of the third type and sometimes of the second type. When clinical symptoms are present, usually they are represented by Raynaud's phenomenon, arthralgias, urticaria, purpura and liver involvement. However, the presence of cryoglobulins in a patient with CTD often does not correlate with clinical picture and other laboratory findings. Eight of our 28 cases (15 SLE and 13 RA) showed third type cryoglobulinemias (7 IgM-IgG and one IgM-IgG-Clq) with a remarkable decrease of serum C4 levels.

Interference of levamisole with inhibition of E-rosette formation by Hodgkin's disease and systemic lupus erythematosus cytotoxic sera.

A new system has been used to test the influence of levamisole on T-cell function. Evidence has been produced that prior exposure to the drug "protects" normal human peripheral blood lymphocytes from the inhibition that cytotoxic sera from patients with Hodgkin's disease and systemic lupus erythematosus exhibit on their E-rosette-forming capacity. Also, damage of this T-cell function already induced by Hodgkin's sera may be partially corrected.

[Histotypes HL-A in the Ligurian population of the islet San Pietro in Sardinia]. / Gli istotipi HLA nella popolazione di origine ligure dell'isola di San Pietro in Sardegna.

The frequency of 17 HLA antigens of loci A and B has been studied by microlymphocytoxicity test in 114 subjects of islet S. Pietro, very close (a few miles) to South-west of Sardinia, but genetically very different from it, deriving the population from a region (Liguria) of continental Northern Italy. The data have been compared with those of a village of Southern Sardinia and with those of a population living in Liguria (continental Italy). The results confirm the striking genetical difference between people of the two islands (S. Pietro and Sardinia) and the close similarity between S. Pietro and Liguria from the genetical point of view.

HLA antigens and erythrocyte glucose-6-phosphate dehydrogenase (G-6-PD) deficiency in Sardinia.

The frequency of 17 HLA antigens from locus A and locus B has been evaluated by microlymphocytotoxicity test in a population of 233 subjects (157 normal and 76 with deficiency of glucose-6-phosphate dehydrogenase [G-6PD]in red cells) from a village of Sardinia, a mediterranean island relatively close to continental Italy. It appears that G-6-PD-deficient people show a frequency of some HLA antigens (A2, A10, B12, BW35) significatively different from normal Sardinian subjects but close to (A10, BW35) or higher (A2, B12) than that of subjects from peninsular Italy.

[Presence of alpha-2-macroglobulin on the membrane of peripheral human blood lymphocytes : a new lymphocyte maker]. / Presenza di alfa-2-macroglobulina sulla membrana di linfociti periferici umani: un nuovo marcatore linfocitario

The presence of alpha-2-macroglobulin (alpha-2-M) on the surface of peripheral blood lymphocytes from normal human subjects and from patients with chronic lymphatic leukaemia (CLL) and with ataxia-teleangectasia was studied by indirect immunofluorescence technique. Same experiments were done on purified subpopulations of normal blood peripheral lymphocytes (B and T). The percentage of alpha-2-M bearing lymphocytes is 17 +/- 6% as regard to normal subjects (Ig-bearing cells are 14%): in CLL the percentage of alpha-2-M bearing cells generally is significantly lower than that of Ig-bearing cells (IgM-IgD). On selected subpopulations, 90% of alpha-2-M bearing cells are present among non T-cells and only 5% among T-cells (probably due to not absolute purification). Blocking experiments using anti-Ig sera did not affect significantly the percentage of alpha-2-M bearing cells and using anti-alpha-2-M sera did not affect that of Ig-bearing cells. The percentage of E-rosette forming cells is not affected by pretreatment of peripheral lymphocytes with anti-Ig and/or anti-alpha-2-M sera. Hypothesis is set forth that alpha-2-M bearing cells could be a lymphocytes subpopulation made by a subgroup of B-cells and by K-cells, taking part in that immunoregulatory system in which collaborate many serum alpha-globulins and T-suppresor cells.

[Comparison between anti-lymphocyte antibodies in systemic lupus erythematosus and in Hodgkin's disease]. / Confronto tra anticorpi antilinbfocitari presenti nel lupus eritematoso sistemico e nella malattia di Hodgkin

Anti-lymphocyte antibodies of cold-reactive IgM-type in Systemic Lupus Erythematosus (SLE) and IgG-type in Hodgkin's disease (HD) by means of immunofluorescent technique have been demonstrated. The contemporaneous attachment of SLE and HD sera on selected peripheral lymphocytes demonstrate that the target cell is T-lymphocyte and that the two kinds of antibodies coat the same cell. Co-capping experiments have shown that the phenomenon has the same behaviour as regard to SLE and HD antibody, indicating that the antigen is the same. Hypothesis about the meaning of these antibodyies have been made.

Anti-lymphocyte antibodies in systemic lupus erythematosus and in Hodgkin's disease: a comparison by immunofluorescence.

Anti-lymphocyte antibodies of IgM type (cold reactive) in systemic lupus erythematosus and of IgG type (not cold reactive) in Hodgkin's disease have been demonstrated by immunofluorescence to attach the same target cell (T-lymphocyte.) The same behaviour of both kinds of antibodies in "co-capping" experiments confirm that the antigen on the cell surface is the same. Hypothesis about the meaning of this phenomenon are made.