Solubilization of proteins from human lymph node tissue and two-dimensional gel storage

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8 M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT (dithiothreitol) and 0.2% carrier ampholytes; (b) 5 M urea, 2 M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N,N-dimethyl-3-ammonio-1-propanesulfonate), 40 mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7 M urea, 2 M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.     

Italian palaeolithic and mesolithic burials

More than 50 skeletons have been found in Italy in Upper Palaeolithic and Mesolithic burials, most of them being of Pleistocene age. As for as Upper Palaeolithic is concerned, two distinct phases are recognized, with a gap in records between them of several millennia: 1) a first phase, in which rich burials, generally of adult or adolescent males, are found. Associated industries are of Gravettian — Early Epigravettian type. Strict rules are followed in funerary practices. Sites of this period are only a few in Italy, but most of them yelded burials; 2) a second phase, of late Würm IV age, with burials found in several sites all over Italy, associated with Final Epigravettian industries. Women as well as children are now buried in single graves, but men still outnumber women. Grave goods are scarce or lacking, except at Arene Candide, where very rich burials, customarily attributed to Mesolithic, but actually of Dryas III age, were found. Truly Mesolithic sepultures are only a few, and burial goods are scarce or lacking.     

A case of chondrodystrophic dwarfism in the Italian late Upper Paleolithic

During excavations by P. Graziosi at Romito Cave near Papasidero (Cosenza) in Calabria, Italy, two skeletons (Romito 1 and 2) were discovered in a common grave. Although the specimens were briefly described by Messeri (Atti X Riuniuone Scientifica Ist. Ital. Preist. Protost., pp. 301-307, 1966), it is generally unknown in the anthropological literature that one of these individuals is a chondrodystrophic dwarf. As such, the specimen provides the earliest known case of dwarfism in the human skeletal record, extending the time span of this genetically determined growth abnormality to approximately 10,000 years ago. The specimen (Romito 2) exhibits features typical of chondrodysplasia, including a high domed skull, compressed cranial base, and in the postcranial skeleton extremely shortened diaphyseal lengths. The unique combination of these features suggests this pathological condition is acromesomelic dysplasia. Besides providing evidence for a greater antiquity of dwarfism than previously known, the fact that this individual reached late adolescence attests to tolerance of Upper Paleolithic groups for severely abnormal individuals and their ability to support members who were of limited economic value to the social group.