Automatic morphometric analysis of retrograde changes in the nucleus n. facialis at different ontogenetic stages in the rat

Summary The grey level index (= GLI) and the fresh volume were determined with the image analyser Micro-Videomat for the nucl. n. facialis after axotomy of the left n. facialis. The experiments were performed on 10 rats in different stages of ontogenesis. The GLI is a quantitative parameter which could be quickly obtained and which demonstrated quantitative changes during retrograde reaction in the respective centres. A decrease in the fresh volumes of the affected nucl. n. facialis could also be demonstrated. The meaning of GLI is discussed.Supported by Deutsche Forschungsgemeinschaft grants Kr 289/10 and Zi 192/1Dedicated to Prof. Dr. H. Leonhardt on the occasion of his 60th birthday     

Mutational analysis of cell cycle arrest, nuclear localization and virion packaging of human immunodeficiency virus type 1 Vpr.

Human immunodeficiency virus type 1 Vpr is a virion-associated, regulatory protein that is required for efficient viral replication in monocytes/macrophages. The protein is believed to act in conjunction with the Gag matrix protein to allow import of the viral preintegration complex in nondividing cells. In cells, Vpr localizes to the nucleus. Recently, we showed that Vpr prevents the activation of p34cdc2-cyclin B. This results in arrest of Vpr-expressing cells in the G2/M phase of the cell cycle. Here, we use a panel of expression vectors encoding Vpr molecules mutated in the amino-terminal alpha-helical region, the central hydrophobic region, or the carboxy-terminal basic region to define the functional domains of the protein. The results showed cell cycle arrest was largely controlled by the carboxy-terminal basic domain of the protein. In contrast, the amino-terminal alpha-helical region of Vpr was required for nuclear localization and packaging into virions. The carboxy terminus appeared to be unnecessary for nuclear localization. In the alpha-helical region, mutation of Ala-30 to Pro resulted in a protein that localized to the cytoplasm. Surprisingly, fusion of Vpr to luciferase resulted in a molecule that failed to localize to the nucleus. In addition, we show that simian immunodeficiency virus Vpr, but not Vpx, induces G2 arrest. We speculate that Vpr has two sites for interaction with cellular factors: one in the alpha-helical region that specifies nuclear localization and one in the carboxy-terminal domain that is required for Cdc2 inhibition.     

Human immunodeficiency virus type 1 viral protein R (Vpr) arrests cells in the G2 phase of the cell cycle by inhibiting p34cdc2 activity.

The Vpr accessory gene product of human immunodeficiency virus types 1 and 2 and simian immunodeficiency virus is believed to play a role in permitting entry of the viral core into the nucleus of nondividing cells. A second role for Vpr was recently suggested by Rogel et al. (M. E. Rogel, L. I. Wu, and M. Emerman, J. Virol. 69:882-888, 1995), who showed that Vpr prevents the establishment in vitro of chronically infected HIV producer cell lines, apparently by causing infected cells to arrest in the G2/M phase of the cell cycle. In cycling cells, progression from G2 to M phase is driven by activation of the p34cdc2/cyclin B complex, an event caused, in part, by dephosphorylation of two regulatory amino acids of p34cdc2 (Thr-14 and Tyr-15). We show here that Vpr arrests the cell cycle in G2 by preventing the activation of the p34cdc2/cyclin B complex. Vpr expression in cells caused p34cdc2 to remain in the phosphorylated, inactive state, p34cdc2/cyclin B complexes immunoprecipitated from cells expressing Vpr were almost completely inactive in a histone H1 kinase assay. Coexpression of a constitutively active mutant p34cdc2 molecule with Vpr relieved the G2 arrest. These findings strongly suggest that Vpr arrests cells in G2 by preventing the activation of the p34cdc2/cyclin B complex that is required for entry into M phase. In vivo, Vpr might, by preventing p34cdc2 activation, delay or prevent apoptosis of infected cells. This would increase the amount of virus each infected cell produced.     

Respiratory gas monitoring in patients with chronic respiratory failure during intensive positive pressure ventilation vs aerosol therapy

Summary The aim of this study is to evaluate the variations of respiratory gas concentration with transcutaneous blood gas measurements (tcm PO₂; tcm PCO₂) during intermittent positive pressure ventilation (I.P.P.V.) and aerosol therapy in patients (5 female, 5 male; 43–75 years) with chronic obstructive pulmonary diseases (COPD) with haemo gas analysis values PO₂<76 mmHg. The subjects underwent transcutaneous blood gas monitoring, 16 minutes during therapy (I.P.P.V. or aerosol) and 14 minutes of recovery. ANOVA analysis of variance was used for statistical analysis. During I.P.P.V. tcm PO₂ increase from the base value (60.6±9 mmHg) already after 2 minutes until the 16th minute (69.7±9); 2 minutes after the end of I.P.P.V. tcm PO₂ falls below the basic one (57.6±7) and then exceeds the basic value until the 14th minute (63.5±10) (p=ns). The base tcm PCO₂ (41.2±7) decreases reaching the maximum decrement at the 16th minute of therapy (32±6.6); at the end of I.P.P.V. it increases without exceeding the base value (39.5±7) (p<0.01). During aerosol therapy, tcm PO₂ increases from the basic one (62.4±10) (maximum after 16 minutes 70±9.5), then it decreases (64±11) and increases againg reaching the maximum growth after 14 minutes of recovery (65.7±11) (p=ns). The tcm PCO₂ values show a decrement below the base value (42.7±4.5) reaching its maximum at the 16th minute of therapy (38.8±7); at the end of aerosol the tem PCO₂ increases (42.5±4.6) (p=ns). The tcm PO₂ variations (I.P.P.V. vs aerosol) did not show any significant statistical difference. At the end of the therapies the tcm PO₂ falls a little more after I.P.P.V. than after aerosol (57.6±7.3 vs 64±11, p=ns); the tcm PCO₂ base values are similar during the two therapies (41.2±7 vs 42.7±4.5, p=ns), but the tcm PCO₂ decrements reach a significant statistical difference after 8–10–12–16 minutes (p<0.05); during the recovery time after 2 minutes (34±6.5 vs 40±6.3, p<0.05) and this difference disappeared at the last recording. We calculate the tcm PO₂ increment and the tcm PCO₂ decrement from the base value. The tcm PO₂ increment is higher during I.P.P.V. than during aerosol therapy. The tcm PCO₂ decrement shows how the tcm PCO₂ falls during I.P.P.V. and how these values remain below the base one until the end of the rest time. Data reported here demonstrate that mechanical physiokinesi therapy improves COPD ventilation so that the respiratory pattern could be ameliorated and preserved.     

Exploring the homogeneity of terrestrial subterranean communities at a local spatial scale

1. Although caves are generally perceived as isolated habitats, at the local scale, they are often interconnected via a network of fissures in the bedrock. Accordingly, caves in close proximity are expected to host the same, or very similar, animal communities. 2. We explored the extent to which subterranean arthropod communities are homogenous at a local spatial scale of less than 1 km², along with which cave-specific environmental features result in a departure from the expected homogeneous pattern. We approached this question by studying richness and turnover in terrestrial invertebrate communities of 27 caves in a small karst massif in the Western Italian Alps. 3. Specialised subterranean species were homogeneously distributed among caves and were not influenced by seasonality. The only factor driving their distribution was the distance from the cave entrance, with deeper caves yielding a greater diversity of species. 4. Significant spatio-temporal turnover in species not specialised to subterranean life was observed. In summer, there was a significant homogenisation of the community and a more even distribution of species among sites; in winter, these species were missing or found exclusively at greater depths, where environmental conditions were more stable. Furthermore, caves at lower elevations yielded, on average, a greater diversity and abundance of these species. 5. This study demonstrated that the theoretical expectation of no turnover in community composition in caves in close proximity is not always met. Turnover can be mostly attributed to seasonal patterns and sampling depth; thus, our findings have implications for planning sampling and monitoring activities in caves.     

Test Based on Independent,but Non-identically Distributed Random Variables

The binomial test is applied for the problem of testing a hypothesis based on a sample of independent, but non-identically distributed random variables. The used basic idea is that each random variable indicates the presence of the hypothesis. Hence each random variable is transformed such that the binomial test can be used as a simple procedure.     

The role of the fluid phase in the viscous response of bovine periodontal ligament

The mechanical response of the periodontal ligament (PDL) is complex. This tissue responds as a hyperelastic solid when pulled in tension while demonstrating a viscous behavior under compression. This intricacy is reflected in the tissue's morphology, which comprises fibers, glycosaminoglycans, a jagged interface with the surrounding porous bone and an extensive vascular network.In the present study we offer an analysis of the viscous behavior and the interplay between the fibrous matrix and its fluid phase.Cylindrical specimens comprising layers of dentine, PDL and bone were extracted from bovine first molars and affixed to a tensile-compressive loading machine. The viscous properties of the tissue were analyzed (1) by subjecting the specimens to sinusoidal displacements at various frequencies and (2) by cycling the specimens in ‘fully saturated’ and in ‘partially dry’ conditions. Both modes assisted in determining the contribution of the fluid phase to the mechanical response.It was concluded that: (1) PDL showed pseudo-plastic viscous features for cyclic compressive loading, (2) these viscous features essentially resulted from interactions between the porous matrix and unbound fluid content of the tissue. Removing the liquid from the PDL largely eliminates its damping effect in compression.