Electron microscopic cytochemistry of adenylyl cyclase activity in mouse spermatozoa.

We investigated adenylyl cyclase activity of mouse spermatozoa by electron microscopic cytochemistry. Subcellular localization of enzyme activity was determined in the presence and absence of bicarbonate ions. Results confirm the existence in sperm of a bicarbonate-regulated adenylyl cyclase, which suggests microdomain signaling.     

Agroforestry management and phytoseiid communities in vineyards in the South of France

This study deals with the long-term effect of agroforestry management (trees within vine crops) on communities of phytoseiid mites. Several plots were considered: vineyards co-planted with Sorbus domestica or Pinus pinea, monocultures of vines and monocultures of S. domestica or P. pinea. All vine plots included two vine cultivars, Syrah and Grenache. Phytoseiid mites have been surveyed in these plots during several years within the previous 10 years. In 2010, samplings were again carried out in these same plots, from May to September, twice a month. Significantly higher densities of Phytoseiidae were observed on the cultivar Syrah (0.85 phytoseiids per leaf) than on Grenache (0.26 phytoseiids per leaf). Furthermore, significantly higher phytoseiid mite densities were observed in the monocultural grapevine plot than in the two co-planted ones. The main species found was Typhlodromus (Typhlodromus) exhilaratus in all vine plots considered. However, Kampimodromus aberrans was observed in the grapevine plots co-planted with the two trees, but never in the monocultural vine plot. Surprisingly, this phytoseiid species was not found on the co-planted trees, nor in the neighbouring uncultivated vegetation. Several hypotheses are discussed to explain such an unexpected distribution. Furthermore, contrary to what has been observed previously, agroforestry management did not seem to favour phytoseiid mite development, especially on the Grenache cultivar. Again, some hypotheses are developed to explain such observations and density modifications.     

An efficient and highly selective deprotection of N-Fmoc-alpha-amino acid and lipophilic N-Fmoc-dipeptide methyl esters with aluminium trichloride and N,N-dimethylaniline.

A novel procedure for the deprotection of the carboxyl group of amino acid methyl esters is presented. The process is carried out by the reagent system aluminium trichloride/N,N-dimethylaniline that can successfully be applied to unblock the carboxyl moiety either of N-Fmoc-protected amino acid methyl esters and N-Fmoc-protected short dipeptide methyl esters. The chiralities of the optically pure amino acid or peptide precursors are maintained totally unchanged.     

CD133 and CD44 cell surface markers do not identify cancer stem cells in primary human gastric tumors

Emerging evidence suggests that tumors contain and are driven by a cellular component that displays stem cell properties, the so-called cancer stem cells (CSCs). CSCs have been identified in several solid human cancers; however, there are no data about CSCs in primary human gastric cancer (GC). By using CD133 and CD44 cell surface markers we investigated whether primary human GCs contain a cell subset expressing stem-like properties and whether this subpopulation has tumor-initiating properties in xenograft transplantation experiments. We examined tissues from 44 patients who underwent gastrectomy for primary GC. The tumorigenicity of the cells separated by flow cytometry using CD133 and CD44 surface markers was tested by subcutaneous or intraperitoneum injection in NOD/SCID and nude mice. GCs included in the study were intestinal in 34 cases and diffuse in 10 cases. All samples contained surface marker-positive cells: CD133(+) mean percentage 10.6% and CD133(+)/CD44(+) mean percentage 27.7%, irrespective of cancer phenotype or grade of differentiation. Purified CD133(+) and CD133(+)/CD44(+) cells, obtained in sufficient number only in 12 intestinal type GC cases, failed to reproduce cancer in two mice models. However, the unseparated cells produced glandular-like structures in 70% of the mice inoculated. In conclusion, although CD133(+) and CD133(+)/CD44(+) were detectable in human primary GCs, they neither expressed stem-like properties nor exhibited tumor-initiating properties in xenograft transplantation experiments.     

Effects of the purified IGF-I complex on the capacitation and acrosome reaction of rabbit spermatozoa

A protein complex containing IGF-I, purified from rabbit seminal plasma, was used to investigate its effects on the capacitation and acrosome reaction of rabbit spermatozoa. Uncapacitated sperm (Pattern F), capacitated sperm (Pattern B), and acrosome-reacted sperm (Pattern AR) were determined by CTC staining, and the results were validated by PSA-staining. The addition of the IGF-I complex to the capacitative medium directed the spermatozoa to spontaneous acrosome reaction. On the other hand, IGF-I complex, added to capacitated spermatozoa, acted as inducer of the acrosome reaction. Results of IVF experiments showed high rates of fertilization with capacitated spermatozoa, acrosome-reacted by either A23187 or IGF I complex, whereas significantly lower rates were obtained with spermatozoa capacitated in the presence of IGF-I complex.