Ex Vivo Reconstruction of the Epidermis With Melanocytes and the Influence of UVB

To study pigmentation, we have reconstructed an epidermis ex vivo with keratinocytes and melanocytes. Keratinocytes and melanocytes were grown first in primary cocultures and separately in secondary cultures, then seeded on a dead deepidermized dermis (Pruniéras type) at a 1:20 melanocyte/keratinocyte ratio. Reconstructed epidermis were grown in a special medium enriched with calcium and fetal bovine serum lifted for 15 days at the air-liquid interface. Using histology, immunohistochemistry and electron microscopy we have shown an excellent level of differentiation of the reconstructed epidermis and a physiologic distribution of dendritic melanocytes in the basal layer capable of melanosome transfer to keratinocytes. UVB irradiation 0.15 J/cm²× 5 consecutive days increased melanocyte numbers and stimulated pigmentation as evidenced macroscopically and microscopically and at the biochemical level. Following UVB irradiation melanosome transfer was markedly increased and isolated or clumps of melanosomes were seen in the basal layers as well as in the stratum corneum. This model allows the study of the physiology of pigmentation ex vivo.     

Characterization of Seed Protein Fractions from Castanea spp

Seed proteins of Castanea sativa and C. crenata were extractedby sequential procedures. Albumins accounted for over 20% ofthe total protein. Globulins were the main storage proteinsand represented close to half of the total protein. Additionof 2-mercaptoethanol was essential for maximum extraction ofthe globulins, a fraction with an amino acid composition similarto that of the 1 IS storage protein from legumes. Negligibleamounts of prolamins (<3%) were present in the seeds. Glutelinsrepresented just under 30% of the total protein. The proteinfractions were characterized by electrophoresis in polyacrylamidegels. The glutelin fraction contained unextracted globulinseven when 2-mercaptoethanol was used. Key words: Castanea spp, seeds, storage protein     

The Proteolytic Potential of Normal Human Melanocytes: Comparison With Other Skin Cells and Melanoma Cell Lines

To understand the contribution of epidermal melanocytes in the proteolytic potential of human skin, we have studied melanocytes grown in a low-serum medium deprived of phorbol esters, cholera toxin, and other non-physiological supplements. We focused on the plasminogen activation system and certain matrix metalloproteinases (gelatinases). Supposing that the proteolytic activity of cells can influence binding to collagen matrix and its reorganization, we have analyzed these parameters as well. We found that human melanocytes secreted tissue-type plasminogen activator and utilised it to generate cell-bound plasmin. No urokinase-type plasminogen activator was detected in the cultures but its receptor was found in cell extracts. Both the 72 kDa and 92 kDa gelatinases were secreted by the cells and in equal amounts. In addition, melanocytes secreted the wide-spectrum proteinase inhibitor alpha-2-macroglobulin. Melanocytes cast into collagen matrices retained a rounded morphology, did not extend processes, and were unable to contract collagen lattices. As a control, these parameters were investigated in parallel in cultures of human keratinocytes, dermal fibroblasts, and two melanoma cell lines. The obtained characteristics suggest that normal human melanocytes are proteolytically active cells. This function may pertain to skin physiology and pathophysiology.     

Effects of fire frequency on survival,growth and fecundity of Paspalum quadrifarium (Lam.) in a grassland of the Flooding Pampa (Argentina)

Abstract Fire and grazing are important factors in the regulation of the grassland structure of the Flooding Pampa (Buenos Aires Province, Argentina dominated by Paspalum quadrifarium plants (henceforth ‘pajonal’)). Winter burning of pajonal is a practice that increases P. quadrifarium growth and its nutritious value for cattle. The objective of this work was to determine the responses of different demographic variables of P. quadrifarium growing in a grassland managed with different fire frequencies. The work was carried out in a pajonal situated in San Ignacio (Ayacucho county, Buenos Aires Province, Argentina). The treatments were: high fire frequency (0.8 fires per year) and low fire frequency (0.4 fires per year). The population of P. quadrifarium was affected by fire frequency. Biomass, number of tillers, and the relative production of tillers per plant did not change with fire frequency. Plant height was negatively affected by fire frequency. Whereas no plant mortality was observed in high fire frequency, 30% of the mature plants died at the end of the experiment in low fire frequency. In both treatments, plant mortality was size‐dependent and juvenile plants were more affected than mature ones. Burning had an immediate negative effect on mature and juvenile plants in terms of fecundity, survival of tillers and basal cover. Tiller survival and fecundity were significantly lower in burned plants than in unburned ones. In the high fire frequency treatment the number of ramets and/or genets and basal cover were greater than in the low fire frequency treatment.     

Seed morphology in Veronica L. (Scrophulariaceae) from south-west Spain

Seeds of 12 species of Veronica from south-west Spain were examined with the scanning electron microscope and stereomicroscope. Based on seed shape, colour, cell shape and surface ornamentation, seven morphological types are recognized. Seeds of the different types are described, illustrated and compared. The distribution of seed types among taxa is given. A key to distinguish the different species or groups of species is provided.     

Morphological and Anatomical Studies of Linaria Species from South-west Spain: Seeds

Seeds of 24 taxa of Linaria from south-west Spain were examinedwith a scanning electron microscope and light microscope. Basedon seed features such as shape, surface ornamentation and cellshape, seven morphological types were recognized. Seeds of thedifferent types are described, illustrated and compared. A keyto distinguish the different species or groups of species isprovided. Copyright 1999 Annals of Botany Company Seed-coat, SEM, anatomy, Scrophulariaceae, Antirrhineae, Linaria .     

OPTIMIZATION OF THE ENUMERATION OF TOTAL AEROBIC BACTERIAL FLORA IN THE FLESH OF SEAFISH

Abstract The total aerobic flora of seafish flesh is weakly halophilic, and requires on average 1.38% NaCl according to statistical studies. Enumeration is optimal on tryptone soya agar or on NaCl supplemented plate count agar (-H₂S), incubated at 20 and 25C, respectively. Plate count agar (-H₂S) was selected because it can also be used for enumeration of hydrogen sulfide-producing bacteria by degradation of sulfur-containing proteins, which are abundant in fish The models employed are sigmoidal. The initial bioburden is too great for there to be a lag phase during storage in ice at 0C. The models show that when the total aerobic microflora count exceeds 100,000 cfu/g, whole or filleted fish stored on ice at 0C are unfit for consumption.