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.     

Immunogenicity and Infectivity of Sporozoites of Mammalian Malaria Isolated by Density-Gradient Centrifugation*

Sporozoites of rodent and simian malaria (Plasmodium berghei and P. cynomolgi) were purified by centrifugation on a linear Renografin/BSA gradient. This procedure made it possible to process rapidly a large number of infected mosquitoes leading to the recovery of a considerable proportion of sporozoites. Gradient-recovered sporozoites (GRS) freed of most bacteria and mosquito tissue contaminants, retained their infectivity and immunogenicity. Mice repeatedly injected i.v. with irradiated GRS of P. berghei acquired total protection against an otherwise lethal sporozoite challenge. GRS of P. berghei and P. cynomolgi induced antisporozoite (CSP) antibody production in rats.     

A REPORT ON VARIATION IN pH AND MICROSCOPICAL COUNT OF BACTERIA IN A PIT OF GRASS SILAGE

SUMMARY: Ignoring wasted material at the surface and sides, the variation in pH in six months old pit silage was smaller from side to side than from top to bottom. At the 2-foot level, where the best quality silage was found, variation was very slight. A correlation was found between pH and the logarithm of the microscopical count of bacteria, the total number of bacteria falling with decreasing pH.     

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.     

A fake Heliconius (Lepidoptera, Nymphalidae)

Four ‘doubtful species’ are listed in catalogues of the genus Heliconius. All are non–existent: (1) H. Carolina (Herbst) is a forgery made from trimmed fragments of at least three butterflies, the two identifiable ones both beingH. erato; (2) H.cinereo–fuscus (Goeze) is a faded specimen of H.ricini; (3) H.arcuatus (Goeze) is so inadequately described that its identity cannot be determined, although it is apparently some species of heliconiine or ithomiine;(4)H.euclea (Godart) is the valid senior name of an ithomiid in the genus Hypothyris.