Rapid Assessment of Maturation Stage and Reproductive Mode in Centrolecytic Eggs of Stick Insects (Phasmatodea) Using DAPI Stain

A rapid three-step DAPI technique is proposed for detecting meiotic stages and sperm head evolution in yolky, fertilized stick insect eggs, which were difficult to analyze with other methods. Fixed eggs were freed from chorionic envelopes and stained directly in DAPI/PBS solution. After rinsing, eggs were singly squashed in a drop of mounting buffer and examined under a microscope with incident fluorescent illumination. The method was almost uniformly successful, and direct observation of nuclear structures, coupled with fluorometry, allowed easy recognition of bivalents, diads, pronuclei and their DNA content. The DAPI method proposed here appears particularly helpful for investigating unusual reproductive modes in eggs with large amounts of yolk.     

Résultats hémodynamiques et cliniques de la dissection, assistée par lunettes chirurgicales, des veines spermatiques au niveau de l’orifice inguinal interne

Inversion of venous flow is well visualized by colour duplex ultrasound and is the basic haemodynamic feature of varicocele. We tried to assess the value of dissection of branches of the spermatic vein in the internal inguinal ring using an operating microscope with a magnification of 4.5x. 16 patients were operated according to the ultrasound classification proposed by Hirsh et al. [8]. Varicocele was grade III in 2 cases, grade IV in 11 cases and grade V in 3 cases and was situated on the left in 13 patients and on the right in 3 patients. The diagnosis was based on clinical examination and colour duplex ultrasound. The operating time was 25–35 min. The incision was 1.5 to 2 cm long. No short-term complications or long-term hydrocele were observed. No reno-spermatic flow was observed on colour duplex ultrasound after six months. The mean testicular volume was 22 ml before surgery and did not change after the operation. This surgical technique is less invasive than the conventional retroperitoneal or laparoscopic procedures, and the incision is smaller than with the percutaneous technique. It is less radical than conventional microsurgery, but the operating time is shorter, and the surgical wound is smaller, sparing the artery and lymphatic vessels. In conclusion, this operation can be considered to be a good option in the surgical treatment of varicocele in view of its low complication rate, the absence of hydrocele, a minimally invasive procedure and the good haemodynamic results.     

Reoccurring neural stem cell divisions in the adult zebrafish telencephalon are sufficient for the emergence of aggregated spatiotemporal patterns

Regulation of quiescence and cell cycle entry is pivotal for the maintenance of stem cell populations. Regulatory mechanisms, however, are poorly understood. In particular, it is unclear how the activity of single stem cells is coordinated within the population or if cells divide in a purely random fashion. We addressed this issue by analyzing division events in an adult neural stem cell (NSC) population of the zebrafish telencephalon. Spatial statistics and mathematical modeling of over 80,000 NSCs in 36 brain hemispheres revealed weakly aggregated, nonrandom division patterns in space and time. Analyzing divisions at 2 time points allowed us to infer cell cycle and S-phase lengths computationally. Interestingly, we observed rapid cell cycle reentries in roughly 15% of newly born NSCs. In agent-based simulations of NSC populations, this redividing activity sufficed to induce aggregated spatiotemporal division patterns that matched the ones observed experimentally. In contrast, omitting redivisions leads to a random spatiotemporal distribution of dividing cells. Spatiotemporal aggregation of dividing stem cells can thus emerge solely from the cells’ history.

An interdisciplinary study of the rules governing cell divisions in a population of neural stem cells in the zebrafish brain reveals the existence of aggregated spatio-temporal division patterns of rapid cell cycles in stem cells, and shows that these patterns can be explained by a simple agent-based model relying solely on the cells‘ division history.     

Comparing patterns of geographic variation in cave crickets by combining geostatistic methods and Mantel tests

Geographic variation patterns of biological characters and environmental variables are compared by using a procedure employing multivariate analyses, production of contour maps by the kriging method with enclosed validation of estimates, and Mantel tests to assess the significance of comparisons. As biological material we chose a sample of Dolichopoda cave crickets populations from Central-Southern Italy. The kriging technique provides estimates of the interpolation error for each true and estimated point. This profitable feature offers the opportunity to use, with ascertained levels of confidence, the estimated z -scores for further analysis and to compare data collected within the same area, but not exactly coincident in location or number. In such a way, we were able to use for subsequent comparisons by means of Mantel tests the maximum number of data points for all data sets, which originally differed in sampling sites. The interpretation of the contour maps and their statistical comparison suggested that allozymes and epiphallus shape data sets follow the phylogenetic pathways within the Dolichopoda populations, whereas variation in leg elongation is almost entirely under the control of an environmental gradient, synthetically described by the cave temperature.     

Structural features of the pore formed by Staphylococcus aureusα-toxin inferred from chemical modification and primary structure analysis

Abstract Staphylococcus aureus α-toxin makes cells and model membranes permeable to ions and uncharged molecules by opening oligomeric pores of uniform size. Its primary sequence reveals peculiar features which give some hints on the structure of the pore. A flexible region separating the toxin into two halves, several amphiphilic β-strands and two amphiphilic α-helices long enough to span the hydrophobic core of the lipid bilayer are predicted. In analogy to bacterial porins, we propose that the inner walls of the pore are, at least in part, built by an amphiphilic β-barrel. The model is consistent with circular dichroism data and with the electrophysiological properties of the pore. Functional information on this toxin were obtained by chemical modification of its four histidine residues. Specific carbethoxylation suggested they have different roles: one is required for specific receptor binding, one for oligomerisation and two for unspecific lipid binding. A tentative assignment of each histidine to its specific role is done on the basis of the structural predictions. A functionally related hemolysin, Aeromonas hydrophyla aerolysin, reveals remarkably similar features including the presence and location of histidines involved in receptor binding and oligomerisation.     

Occurrence of radon in some schools of Parma and Reggio Emilia (Northern Italy)

Summary Forty-seven measurement of radon concentration were made in some schools of Parma, Reggio Emilia, Albinea and Borzano. The method used was that of activated carbon canisters, which were placed in classrooms, laboratories, libraries and headmaster's offices for at least 48 hours in the period November '90–March '91. It was possible to determine the amount of radon in each canister counting the Pb-214 and Bi-214 gamma emitters by means of NaI (Tl) and Ge (I) detectors. The mean radon concentrations were: 20 Bq/m³ in Parma; 24 Bq/m³ in Reggio Emilia; 46 Bq/m³ in Borzano and 52 Bq/m³ in Albinea. The values recorded in schools are similar to the values previously recorded in dwellings of Parma and Reggio Emilia.