Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization

A procedure is described which permits the isolation from the prepuberal mouse testis of highly purified populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene primary spermatocytes, leptotene and zygotene primary spermatocytes, pachytene primary spermatocytes and Sertoli cells. The successful isolation of these prepuberal cell types was accomplished by: (a) defining distinctive morphological characteristics of the cells, (b) determining the temporal appearance of spermatogenic cells during prepuberal development, (c) isolating purified seminiferous cords, after dissociation of the testis with collagenase, (d) separating the trypsin-dispersed seminiferous cells by sedimentation velocity at unit gravity, and (e) assessing the identity and purity of the isolated cell types by microscopy. The seminiferous epithelium from day 6 animals contains only primitive type A spermatogonia and Sertoli cells. Type A and type B spermatogonia are present by day 8. At day 10, meiotic prophase is initiated, with the germ cells reaching the early and late pachytene stages by 14 and 18, respectively. Secondary spermatocytes and haploid spermatids appear throughout this developmental period. The purity and optimum day for the recovery of specific cell types are as follows: day 6, Sertoli cells (purity>99 percent) and primitive type A spermatogonia (90 percent); day 8, type A spermatogonia (91 percent) and type B spermatogonia (76 percent); day 18, preleptotene spermatocytes (93 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent).     

Biometric variations and oxidative stress responses in juvenile Clarias gariepinus exposed to Termex®

The current study investigated the effects of termite insecticide, Termex® (imidacloprid 35.50% SC), on biometric variations and oxidative stress biomarkers in Clarias gariepinus. Fish were exposed to 4.00 and 6.00 µg l^–1 sublethal Termex® concentrations in 2017. The gill and liver tissues were sampled on days 7, 14, 21 and 28 and the results indicated that hepatosomatic index (HSI) decreased significantly when compared with the control on days 14, 21 and 28. The condition factor (CF) and viscera-somatic index (VSI) also decreased during the study period. The decrease was greater at 6.00 µg l^–1 Termex® concentration on days 21 and 28 for CF and days 14 to 28 for VSI, respectively. The lipid peroxidation (LPO) in both tissues was highest in the 6.00 µg l^?1 Termex® and increased with the duration. There was significant decrease (p < 0.05) in superoxide dismutase and glutathione peroxidase values, but significant increase in catalase activity in both tissues. The values of glutathione reductase in both tissues were comparable to the control, except on days 21 and 28 in the liver. There was negative correlation between the LPO in tissues and the HSI, CF and VSI values. The use of Termex® in the environment should be monitored to safeguard the health of aquatic organisms.     

Relationship between the secretor status and the expression of ABH blood group antigenic determinants in human intestinal brush-border membrane hydrolases

At least six hydrolases of the human intestinal brush-border membrane bear ABH blood group antigenic determinants related to the erythrocyte phenotype: the intestinal glycoproteins of blood group A and B subjects express A or B determinants, respectively, while blood group O subjects express the H determinant identified with Ulex europaeus lectin I. These expressions are under the control of the secretor gene: ABH antigens were not detected in the hydrolases of non-secretor subjects.     

Phylogenetic utility of elongation factor-1 alpha in noctuoidea (Insecta: Lepidoptera): the limits of synonymous substitution

To test its phylogenetic utility, nucleotide sequence variation in a 1,240-bp fragment of the elongation factor-1 alpha (EF-1 alpha) gene was examined in 49 moth species representing the major groups of the superfamily Noctuoidea. Both parsimony and distance analyses supported the monophyly of nearly all groups for which there are clear morphological synapomorphies. Clades of subfamily rank and lower, probably mid-Tertiary and younger, were strongly supported. The third codon position contains 88% of variable sites, and approaches saturation at approximately 20% sequence divergence, possibly due to among-site rate heterogeneity and composition bias; higher divergences occur only in association with shifts in composition. Surprisingly, the few nonsynonymous changes appear no more phylogenetically reliable than synonymous changes. Signal strength for basal divergences is weak and fails to improve with character weighting; thus, dense taxon sampling is probably needed for strong inference from EF-1 alpha regarding deeper splits in Noctuoidea (probably early Tertiary). EF-1 alpha synonymous changes show promise for phylogeny reconstruction within Noctuidae and other groups of Tertiary age.      

Molecular phylogeny of the major arthropod groups indicates polyphyly of crustaceans and a new hypothesis for the origin of hexapods

A phylogeny of the arthropods was inferred from analyses of amino acid sequences derived from the nuclear genes encoding elongation factor-1 alpha and the largest subunit of RNA polymerase II using maximum- parsimony, neighbor-joining, and maximum-likelihood methods. Analyses of elongation factor-1 alpha from 17 arthropods and 4 outgroup taxa recovered many arthropod clades supported by previous morphological studies, including Diplopoda, Myriapoda, Insecta, Hexapoda, Branchiopoda (Crustacea), Araneae, Tetrapulmonata, Arachnida, Chelicerata, and Malacostraca (Crustacea). However, counter to previous studies, elongation factor-1 alpha placed Malacostraca as sister group to the other arthropods. Branchiopod crustaceans were found to be more closely related to hexapods and myriapods than to malacostracan crustaceans. Sequences for RNA polymerase II were obtained from 11 arthropod taxa and were analyzed separately and in combination with elongation factor-1 alpha. Results from these analyses were concordant with those derived from elongation factor-1 alpha alone and provided support for a Hexapoda/Branchiopoda clade, thus arguing against the monophyly of the traditionally defined Atelocerata (Hexapoda + Myriapoda).      

The development of the grape berry cuticle in relation to susceptibility to bunch rot disease

Some physical and morphological factors of grape berry cuticlewere investigated at different developmental stages of threeclones ofVitis vinifera cv. Pinot noir. The surface morphologyof grape berries was examined by scanning electron microscopyand cuticle anatomy was examined by light and transmission electronmicroscopy. During the period from flowering to maturity, thecomposition of the cuticular waxes changed, corresponding withan increase of waxy deposits and significant modifications ofthe wax surface morphology. The content in cutin per unit surfacedecreased more than 2.5-fold between berry set (16 d after anthesis)and veraison of the grape berries, and might predispose thegrape berry to fungal infection. This result was correlatedwith the differentiation of the cuticle layers and particularlywith a decrease in the thickness of the primary cuticle at harvest. Key words: Botrytis cinerea, cuticle, cutin, epicuticular waxes, Vitis vinifera L