Fission Yeast rad12+ Regulates Cell Cycle Checkpoint Control and Is Homologous to the Bloom’s Syndrome Disease Gene

The human BLM gene is a member of the Escherichia coli recQ helicase family, which includes the Saccharomyces cerevisiae SGS1 and human WRN genes. Defects in BLM are responsible for the human disease Bloom’s syndrome, which is characterized in part by genomic instability and a high incidence of cancer. Here we describe the cloning of rad12⁺, which is the fission yeast homolog of BLM and is identical to the recently reported rhq1⁺ gene. We showed that rad12 null cells are sensitive to DNA damage induced by UV light and γ radiation, as well as to the DNA synthesis inhibitor hydroxyurea. Overexpression of the wild-type rad12⁺ gene also leads to sensitivity to these agents and to defects associated with the loss of the S-phase and G₂-phase checkpoint control. We showed genetically and biochemically that rad12⁺ acts upstream from rad9⁺, one of the fission yeast G₂ checkpoint control genes, in regulating exit from the S-phase checkpoint. The physical chromosome segregation defects seen in rad12 null cells combined with the checkpoint regulation defect seen in the rad12⁺ overproducer implicate rad12⁺ as a key coupler of chromosomal integrity with cell cycle progression.     

Schizosaccharomyces malidevorans and Sz. octosporus homologues of Sz. pombe rad9, a gene that mediates radioresistance and cell-cycle progression

The rad9 gene of Schizosaccharomyces pombe is involved in promoting resistance to ionizing radiation and UV light, as well as regulating cell cycle progression after irradiation. We have isolated functional rad9 cognates from two other fission yeasts, Sz. malidevorans and Sz. octosporus, that can restore radioresistance and the radiation-induced G2 delay response to Sz. pombe rad9::ura4 cells. The Sz. pombe and Sz. malidevorans genes are identical at the nucleotide sequence level, which reflects their close evolutionary relationship. Each bears three introns and codes for a 47464-Da protein that contains 426 amino acids (aa). In contrast, Sz. octosporus rad9 contains five introns and codes for a 48210-Da protein that is 432-aa long. The Sz. pombe rad9 product is only 65% identical and 80% similar to the corresponding Sz. octosporus gene product. All of the strains synthesize a rad9 RNA of approx. 1.6 kb. The presence of a rad9-like gene in these yeasts suggests that the cellular process(es) mediated by rad9, and used by these organisms to increase survival and transiently delay cycling in G2 after irradiation, are conserved. The isolation, analyses and comparison of rad9 genes from different organisms should aid in elucidating the specific biological role of the corresponding protein and especially help pinpoint regions important for function.     

Applications of solids NMR to the analysis of insect sclerotized structures

This article reviews the solids NMR research conducted on insect sclerotized structures in the last 10 years and previews some of the experiments that will be conducted in the future. Solids NMR has been used as a noninvasive approach to investigate the chemical compositions of, and some covalent interactions that occur in, several types of sclerotized structures that are otherwise highly intractable to conventional chemical analyses. Sclerotization is a complex process used by insects to confer stability and mechanical versatility to their cuticular exoskeletons and certain other proteinaceous structures. Samples analyzed include cuticular exoskeletons, egg cases, egg shells, cocoons and peritrophic membranes. Cross polarization, dipolar decoupling, magic angle spinning, magnetization dephasing, and isotropic enrichment were used to obtain high resolution spectra that provide information about the types and relative concentrations of carbon atoms as well as internuclear distances and covalent bonds between carbon and nitrogen atoms. Relative amounts of protein, chitin, catechols, lipids, pigment, and oxalate were estimated. Covalent interactions between protein nitrogens and catechol carbons were detected in the stiff brown pupal cuticle of the tobacco hornworm, Manduca sexta. The results of these solids NMR studies support the hypothesis that sclerotization of insect structures occurs primarily when quinones derived from N-acylcatecholamines form cross-links and adducts with functional groups of proteins deposited in the structures. Future applications of solids NMR will utilize advanced techniques for further probing the covalent interactions of ¹³C, ¹⁵N and ¹⁷O-labeled catechols, chitin and protein in sclerotized structures.     

OLFACTORY RESPONSE OF THE GRASSHOPPER MELANOPLUS SANGUINIPES TO PLANT ODOURS AND VOLATILE COMPOUNDS

Abstract Electroantennograms (EAGs1 were recorded from male adults of Melanoplus sanguinipes to foliage odours from 5 plant species and 9 volatile compounds as well as their blends. Higher EAG responses were recorded for Louisianna sage and Alfalfa foliages, compared to other three gramineous plants. Among green odour chelricals, 6-carbon alcohols (e. g. E-2-hexen-1–01 and E-3-hexen-1–01] resulted in the highest EAG responszs. No significant differences among the pentenol, 3-pentanone and blank control were observed. In ternary and quaternary mixture tests of volatile compounds, the combinations with pentenol did not elicit greater amplitudes; thus pentenol and 3-pentanone were proved to be unimportant to the response of this generalist grasshopper. The EAG data are discussed with regard to the role of host plant volatiles in host plant orientation and selection by the grasshopper.     

Wounds incurred in routine cell culture prolong the duration of the life cycle ofAcetabularia acetabulum and require K+ to heal

Summary We have examined the pressure-wound healing response inAcetabularia acetabulum (L.) Silva (Chlorophyta). Commonly incurred in routine cell culture, these wounds induce disruption of the vacuole and translocation of the cytoplasm away from the wound site. Daily wounding of individual cells retarded cytoplasmic healing over time, but had no effect on the rate of membrane healing. The position of the wound along the cell stalk also affected the ability of the cell to heal: cells wounded near the rhizoid healed at least 1.9 times more slowly and were only half as likely to achieve reproduction as were cells wounded either near the apex or at mid-stalk. The 50% mortality of cells wounded at the rhizoid suggests the existence of a physical structure near the primary nucleus which is important to cell viability. The impact of wounding on reproductive potential and time to heal differed with the phase of cell development: juvenile and early adult cells healed 2–2.5 times more quickly but were less likely to achieve reproduction than late adult or reproductive cells. Growth at very high population densities (2.5 cells/ml) impaired the ability of the cells to heal. Growth of cells in seawater containing a range of potassium concentrations revealed that healing depends on potassium and is optimal at a concentration of — 1.51ogM potassium.Abbreviations SE standard error of the mean - LD light/dark - NEM N-ethylmaleimide     

Pulmonary perfusion in the prone and supine postures in the normal human lung.

Prone posture increases cardiac output and improves pulmonary gas exchange. We hypothesized that, in the supine posture, greater compression of dependent lung limits regional blood flow. To test this, MRI-based measures of regional lung density, MRI arterial spin labeling quantification of pulmonary perfusion, and density-normalized perfusion were made in six healthy subjects. Measurements were made in both the prone and supine posture at functional residual capacity. Data were acquired in three nonoverlapping 15-mm sagittal slices covering most of the right lung: central, middle, and lateral, which were further divided into vertical zones: anterior, intermediate, and posterior. The density of the entire lung was not different between prone and supine, but the increase in lung density in the anterior lung with prone posture was less than the decrease in the posterior lung (change: +0.07 g/cm(3) anterior, -0.11 posterior; P < 0.0001), indicating greater compression of dependent lung in supine posture, principally in the central lung slice (P < 0.0001). Overall, density-normalized perfusion was significantly greater in prone posture (7.9 +/- 3.6 ml.min(-1).g(-1) prone, 5.1 +/- 1.8 supine, a 55% increase; P < 0.05) and showed the largest increase in the posterior lung as it became nondependent (change: +71% posterior, +58% intermediate, +31% anterior; P = 0.08), most marked in the central lung slice (P < 0.05). These data indicate that central posterior portions of the lung are more compressed in the supine posture, likely by the heart and adjacent structures, than are central anterior portions in the prone and that this limits regional perfusion in the supine posture.     

Metabarcoding improves detection of eukaryotes from early biofouling communities: implications for pest monitoring and pathway management

In this experimental study the patterns in early marine biofouling communities and possible implications for surveillance and environmental management were explored using metabarcoding, viz. 18S ribosomal RNA gene barcoding in combination with high-throughput sequencing. The community structure of eukaryotic assemblages and the patterns of initial succession were assessed from settlement plates deployed in a busy port for one, five and 15 days. The metabarcoding results were verified with traditional morphological identification of taxa from selected experimental plates. Metabarcoding analysis identified > 400 taxa at a comparatively low taxonomic level and morphological analysis resulted in the detection of 25 taxa at varying levels of resolution. Despite the differences in resolution, data from both methods were consistent at high taxonomic levels and similar patterns in community shifts were observed. A high percentage of sequences belonging to genera known to contain non-indigenous species (NIS) were detected after exposure for only one day.     

Group Prerelease Training Yields Positive Rehabilitation Outcomes Among Juvenile Mantled Howlers (<Emphasis Type="Italic">Alouatta palliata</Emphasis>)

Wildlife rehabilitation presents many challenges, especially when anthropogenic threats yield large numbers of naive juvenile animals. In particular, many primates obtain survival skills such as the identification of palatable foods, avoidance of unsafe substrates, and antipredatory behaviors from conspecifics, and require prerelease training by humans in the absence of opportunities for social learning. Nonetheless, few primate rehabilitation programs quantitatively evaluate the success of prerelease training programs, making the establishment of best practices for many species difficult. Here, we quantitatively assess whether three juvenile mantled howlers (Alouatta palliata) gained independence from humans and increased their frequencies of naturalistic behaviors while participating in a rehabilitation program in Panama that consisted of daily “bush outings” with an operant conditioning component. We evaluated the role of rehabilitating animals in groups in particular, by determining whether the three rehabilitants were more likely to exhibit naturalistic behaviors when in closer proximity to conspecifics. Across the 54-day study period, all three rehabilitants improved significantly in at least two out of the three tested metrics (approaching humans less frequently, traveling higher into the canopy, and spending more time foraging for forest foods). Individuals were more likely to venture into trees and less likely to approach humans when in close proximity to conspecifics, suggesting that group rehabilitation positively impacts the acquisition of naturalistic behaviors by this species. Our results support previous notions of prerelease training as a crucial rehabilitation tool, and highlight the importance of group rehabilitation in programs targeting social animals, even when naturalistic group compositions cannot be approximated.