Pollen Fertility Restoration by Nuclear Gene Fr in Cms Bean: Nuclear-Directed Alteration of a Mitochondrial Population

Two nuclear genes, Fr and Fr2, have been identified that restore pollen fertility to cytoplasmic male sterile (CMS) common bean (Phaseolus vulgaris L.) by apparently distinct mechanisms. Whereas Fr2 appears to suppress the expression of a male sterility associated mitochondrial sequence (designated pvs), Fr restores pollen fertility by causing the elimination of this unusual mitochondrial DNA segment. To further investigate the mechanism of Fr action, Fr and Fr2 were cointroduced into the nucleus of a bean line containing the sterility inducing cytoplasm. When the effect of pvs was suppressed by Fr2, the presence of Fr no longer directed the elimination of the mitochondrial pvs sequence. This result suggests that the Fr function is dependent on proper expression of the pvs sequence. To evaluate the temporal and spatial patterns of Fr action, we undertook a polymerase chain reaction-based approach to trace the fate of the pvs sequence in different tissues of F(2) and F(3) fertile-restored plants derived from a genetic cross between a cytoplasmic male sterile line of common bean, CMS-Sprite (frfr), and fertility restorer line R351 (FrFr). We demonstrate that the Fr-directed disappearance of pvs sequence occurs during flower development. Elimination of the pvs sequence from developing megaspores results in permanent fertility restoration in the following generations. Genetic analysis demonstrated that permanent fertility restoration, that is, the complete elimination of pvs from reproductive tissues requires two doses of the Fr allele or the absence of fr in F(2) individuals. The effect of Fr was reversible until full fertility was achieved. On the basis of these results, we propose a model for the mechanism of pvs elimination by the Fr gene and discuss the dynamics of pvs-containing mitochondrial transmission in the presence of the Fr gene.     

Ultrastructure of Kunjin virus-infected cells: colocalization of NS1 and NS3 with double-stranded RNA, and of NS2B with NS3, in virus-induced membrane structures.

The subcellular location of the nonstructural proteins NS1, NS2B, and NS3 in Vero cells infected with the flavivirus Kunjin was investigated using indirect immunofluorescence and cryoimmunoelectron microscopy with monospecific antibodies. Comparisons were also made by dual immunolabelling using antibodies to double-stranded RNA (dsRNA), the putative template in the flavivirus replication complex. At 8 h postinfection, the immunofluorescent patterns showed NS1, NS2B, NS3, and dsRNA located in a perinuclear rim with extensions into the peripheral cytoplasm. By 16 h, at the end of the latent period, all patterns had changed to some discrete perinuclear foci associated with a thick cytoplasmic reticulum. By 24 h, this localization in perinuclear foci was more apparent and some foci were dual labelled with antibodies to dsRNA. In immuno-gold-labelled cryosections of infected cells at 24 h, all antibodies were associated with clusters of induced membrane structures in the perinuclear region. Two important and novel observations were made. First, one set of induced membranes comprised vesicle packets of smooth membranes dual labelled with anti-dsRNA and anti-NS1 or anti-NS3 antibodies. Second, adjacent masses of paracrystalline arrays or of convoluted smooth membranes, which appeared to be structurally related, were strongly labelled only with anti-NS2B and anti-NS3 antibodies. Paired membranes similar in appearance to the rough endoplasmic reticulum were also labelled, but less strongly, with antibodies to the three nonstructural proteins. Other paired membranes adjacent to the structures discussed above enclosed accumulated virus particles but were not labelled with any of the four antibodies. The collection of induced membranes may represent virus factories in which translation, RNA synthesis, and virus assembly occur.     

Dirofilaria immitis: heartworm infection alters pulmonary artery endothelial cell behavior

Mupanomunda, Maria, Jeffrey F. Williams, Charles D. Mackenzie, and Lana Kaiser. Dirofilaria immitis:heartworm infection alters pulmonary artery endothelial cell behavior.J. Appl. Physiol. 82(2): 389-398, 1997.Thepathogenesis of filariasis has generally been attributed to eitherphysical presence of the adult parasites or the host's immune responseto the parasites. However, the spectrum of filariasis cannot beentirely explained by these causes, and other mechanisms must beoperative. It is now evident that factors released by filarialparasites likely contribute to the pathogenesis of filarial diseases.Adult heartworms (Dirofilaria immitis) reside in the rightheart and pulmonary artery, so the pulmonary artery should be exposedto the highest concentration of filarial factors. We tested thehypothesis that endothelium-dependent relaxation is altered in the invitro pulmonary artery from heartworm-infected dogs. Relaxationresponses to endothelium-dependent vasodilators (methacholine,bradykinin, substance P, and A-23187) and the non-endothelium-dependent vasodilator nitroglycerin and contractile responses were measured inrings of pulmonary artery from control and heartworm-infected dogs.Endothelium-dependent relaxation was assessed in the presence andabsence of inhibitors of nitric oxide synthase, cyclooxygenase, andguanylate cyclase. Responses to methacholine, substance P, and A-23187,but not to bradykinin, nitroglycerin, norepinephrine, or KCl, weredepressed in pulmonary artery from heartworm-infected dogs whencompared with control, suggesting that changes in endothelial cell andnot vascular smooth muscle behavior are involved in altered relaxation.The mechanism of endothelium-dependent relaxation in control pulmonaryartery appears to involve nitric oxide in the case of methacholine andboth nitric oxide and a cyclooxygenase product in the case ofbradykinin and A-23187. The mechanism of endothelium-dependentrelaxation in pulmonary artery from heartworm-infected dogs was notclearly elucidated. These data provide no evidence that heartworminfection globally influences either endothelial cell receptor functionor the vascular smooth muscle guanylate cyclase guanosine 3,5-cyclicmonophosphate system, making it likely that changes in intracellularsignaling are primarily responsible for the observed alteration ofendothelium-mediated relaxation. Alteration of endothelial cellfunction by filarial parasites may be an important component inthe pathology associated with filariasis.

     

Glucan common to the microcyst walls of cyst-forming bacteria.

Chemical analysis indicated that D-glucose is tha major neutral monosaccharide present in the microcysts of a range of gram-negative bacteria. Varying amounts of other neutral sugars were found. The glucose was mainly present as a glucan that could be extracted from microcysts of representative strains with alkali or mild acid treatment. The glucan could be identified as an alpha-1,3-linked polymer on the basis of (i) periodate resistance of the extracted polymer and the material present in microcysts; (ii) lectin agglutination of the microcysts; (iii) lectin precipitation of the extracted glucans; and (iv) susceptibility of the glucan either in the walls or after extraction to a specific alpha-1,3-glucanase from Aspergillus nidulans, yielding glucose as the sole hydrolysis product. The galactosamine found in microcysts of Myxococcus xanthus by other workers is clearly a component of another polymer, distinct from the glucan. The presence of an alpha 1,3-linked glucan, common to microcyst walls of various bacterial genera, probably contributes to the rigidity of the walls of these forms and, inter alia, to their resistance to ultrasonic treatment. Preliminary experiments indicate that the gulcan is discarded on germination of the microcysts rather than being broken down by specific enzymes.     

Secretion of lysosomal enzymes by human placental villi in vitro

Summary Chorionic villi from first trimester and term human placentas have been incubated in vitro and shown to release the lysosomal enzymes, -hexosaminidase, -glucosidase and -gluctlronidase. There was negligible release of the cytoplasmic enzyme, lactate dehydrogenase, under the same conditions. The first trimester villi released proportionally more of their lysosomal enzyme content than did the term villi. Extracellular levels of -hexosaminidase were raised and those of -glucosidase and, -glucuronidase were lowered when tissue was incubated with 1 M colchicine, suggesting that microtubules are involved in the control of lysosomal enzyme release from placental villi.     

Case-control study of recidivist drivers involved in fatal highway accidents in Alberta in 1970-72.

A 3-year enpidemiologic study of fatal motor vehicle accidents was carried out by the Alberta Task Force on Highway Accidents. It was found that 11.1% of culpable drivers had been driving while impaired by alcohol on at least one occasion prior to the fatal accident, while only 3.3% of exonerated drivers had previous records. This significant difference led to a case-control study that demonstrated that the recidivist drivers (case group) differed significantly in a number of aspects from the innocent drivers (control group)--notably, age distribution, sex, whether drinking before the accident, blood alcohol concentration, ethnic distribution, class of accident and type of collision. Thus the recidivist driver is a greater threat to highway safety than the innocent driver and needs special attention by legislators and law enforcement agencies.     

Effects of legumes on soil physical quality in a maize crop

The effect of intercropped legumes and three N fertilizer rates in a continuous maize (Zea mays L.) cropping system on the physical properties of two soils were investigated for three years. The legumes, being a mixture of alfalfa, clover and hairy vetch, had a significant cumulative effect on some physical properties of both soil. The lowest stability and smallest mean weight diameter of soil aggregates were associated with monoculture maize plots. Aggregate size and stability were not affected by N fertilization at any of the rates of 0, 70, and 140 kg ha^-1 in intercropped plots, except that aggregate stability was actually reduced by N fertilization in one soil, the Ste. Rosalie clay. In maize plots in both soils, stability and size of soil aggregates were significantly increased with increased added N. Intercropped legumes significantly decreased dry bulk density and soil penetration resistance. Added N had no measurable influence on these compaction factors. Soil water properties were not significantly affected by either intercropping or N fertilization. Positive effects noted on soil aggregation and other physical properties in intercropped plots are the result of enhanced root activity, or incorporation of legumes as green manure, or both. Improvement of soil structure in maize plots associated with increasing N application was the result of increased maize-root residues.