Proteolysis of milk fat globule membrane proteins in preterm milk: a transient phenomenon with a possible biological role?

Milk fat globule membrane (MFGM) proteins constitute a milk fraction currently of great interest, as they appear to significantly contribute to milk protective role. We investigated these proteins in human preterm colostrum and milk. For the former we found a peculiar 2-DE pattern, with a spot concentration at low molecular weight, which mass spectrometry analysis showed to be fragments belonging to some MFGM proteins with a well-known biological and especially immunological role: lactadherin, membrane-associated lactoferrin, butyrophilin, clusterin and heavy-chain immunoglobulin. Since we were able to rule out protease activity after specimen collection, we hypothesize the localization of the proteolytic enzymes in the alveolar cell membranes of the mammary gland. This mechanism is probably under hormonal control and the unexpected advent of preterm delivery would not allow hormonal conditions typical of lactation to occur immediately, causing a delay in enzymatic inhibition. This hypothesis is supported by some of our results, picturing a peculiar transient phenomenon of adaptation of the mammary-gland-membrane proteins after preterm delivery. Further studies will be required to verify whether the presence of protein fragments exerts a specific biological and immuno-defensive role in preterm infants, thus adding evidence to the outstanding biological role and benefits of mother's own milk in feeding preterm infants.

Media containing aromatic compounds induce peculiar proteins in Acinetobacter radioresistens, as revealed by proteome analysis.

An Acinetobacter radioresistens strain able to grow on phenol or benzoate as sole carbon and energy source through the beta-ketoadipate pathway was isolated in our laboratories. In previous research, we found a different expression of catechol-1,2-dioxygenase isoenzymes (C-1,2-O) depending on the growth substrate (phenol or benzoate). In the present study, we used proteome techniques to extend our investigation to other enzymes involved in the aromatic degradation pathway. Since the first nontoxic metabolite in this route is cis,cis-muconic acid, we focused our attention on the enzymes leading to this compound, chiefly phenol hydroxylase (PH), benzoate dioxygenase (BD), cis-1,2-dihydroxycyclohexa-3,5-diene-1-carboxylate dehydrogenase (D) and C-1,2-O. In particular, the A. radioresistens proteome was monitored under different growth substrate conditions, using acetate, benzoate, or phenol as sole carbon source. We compared the protein maps by software image analysis and detected marked differences, suggesting the inducibility of most enzymes. This research also sought to evaluate the conditions allowing the best expression of enzymes to be used in immobilized systems suitable for bioremediation. The experimental data indicate that benzoate is the best carbon source to gain the highest amount of C-1,2-O and D, while phenol is the best growth substrate to obtain PH.

Additional Ornamental Species as Hosts of Impatiens Necrotic Spot Tospovirus in Italy.

Impatiens necrotic spot (INSV) and tomato spotted wilt (TSWV) tospoviruses are among the most important viral pathogens of glasshouse ornamental plants worldwide (1). Tospovirus infections drastically reduce the market value of plants and create certification problems for international traders. As with TSWV, the number of natural host species recorded for INSV is steadily increasing (2). In 1998, severe INSV infections were found on different ornamental plants in glasshouses in the Piedmont Region of northwestern Italy, together with heavy infestations of the thrips Frankliniella occidentalis. A high proportion of plants were infected with INSV, as shown by enzyme-linked immunosorbent assay (ELISA) with polyclonal antisera and monoclonal antibodies against its nucleocapsid protein. Results were confirmed by sap-inoculation to indicator hosts. Some species were already known to be susceptible to INSV, but others are apparently new hosts (2): Ageratum houstonianum (Asteraceae), showing small necrotic rings and leaf malformation; Cordyline terminalis (Agavaceae), showing chlorotic-necrotic ringspots and leaf malformation; Dianthus chinensis (Caryophyllaceae), showing stunting, mosaic, and leaf malformation (some plants had symptomless infections); Episcia capreata (Gesneriaceae), showing necrotic spots on stems and leaves; Godetia grandiflora (Onagraceae), showing necrotic rings; Maranta leuconeura (Marantaceae), showing chlorotic-necrotic spots and apical malformation; Peperomia obtusifolia (Piperaceae), showing necrotic ringspots and leaf malformations; Scindapsus aureus (Araceae), showing necrotic spots and rings; Torenia fournieri (Scrophulariaceae), showing necrosis on stems and apexes. Thrips feeding damage was high on some species, particularly those showing necrotic ringspot symptoms. It often was difficult to distinguish between true systemic symptoms and local INSV infection at thrips feeding sites. Capsicum sp., Coleus blumei, and Dahlia sp., which also were infected in our study, are species known to be infrequent hosts of INSV (2). References: (1) M. L. Daughtrey et al. Plant Dis. 81:1220, 1997. (2) D. Peters. 1998. Pages 107-110 in: Abstr. 4th Int. Symp. Tospovirus Thrips Floral Vegetable Crops. University of Wageningen, The Netherlands.

cDNA characterization and chromosome mapping of the human GAS2 gene.

Murine Gas2 is a microfilament-associated protein whose expression is increased at growth arrest in mammalian cells. During apoptosis, Gas2 is specifically cleaved at its C-terminus by a still unknown ICE-like protease, and the processed protein induces dramatic rearrangements in the cytoskeleton when overexpressed in several cell types. Here we report the characterization of a cDNA encoding the human homologue of Gas2, showing high conservation with the murine counterpart at the protein level. Fluorescence in situ hybridization analysis and radiation hybrid mapping localized the GAS2 gene on human chromosome 11p14.3-p15.2, in a region homologous to the gas2 region on mouse chromosome 7.

First Report of Moroccan Watermelon Mosaic Potyvirus in Zucchini in Italy.

Unusual symptoms were observed in summer 1997 in field zucchini of several cultivars grown in central Italy. Symptoms included reduction in growth, severe mosaic, blistering and deformation of leaves, and malformation on fruits. Plants gave negative results in enzyme-linked immunosorbent assay (ELISA) for cucumber mosaic cucumovirus, squash mosaic comovirus, papaya ringspot, zucchini yellow fleck, zucchini yellow mosaic, and watermelon mosaic 2 potyviruses. Positive reactions were obtained in ELISA with a monoclonal antibody that reacts with many potyviruses (from J. Vetten, Braunschweig) and with polyclonal antibodies (Sanofi, France) to Moroccan watermelon mosaic potyvirus (MWMV). Field symptoms were reproduced in zucchini cvs. Genovese and Striato d'Italia by mechanical inoculation of samples from symptomatic field plants. Chenopodium amaranticolor, C. quinoa, and Gomphrena globosa gave local lesions, while Citrullus lanatus cv. Crimson Sweet, Cucumis melo cv. Top Mark, C. metuliferus, C. sativus cvs. Marketer, MM76, and Sweet Slice, and Cucurbita maxima reacted with systemic mosaic. C. melo cv. Doublon formed necrotic local lesions followed by systemic necrosis. No infection occurred in Nicotiana benthamiana, N. clevelandii, N. glutinosa, Phaseolus vulgaris cv. Black Turtle, Pisum sativum cv. Alaska, tobacco White Burley, and Vigna unguiculata. These data are in agreement with the known host range of MWMV. MWMV is a tentative species in the genus potyvirus, widely present in Africa and occasionally found in Spain (1,2). Further spread of this virus in the Mediterranean area will create new problems for commercial cucurbit production and breeding, and for diagnosis. References: (1) E. Kabelka and R. Grumet. Euphytica 95:237, 1997. (2) N. M. McKern et al. Arch. Virol. 131:467, 1993.