Markers for trans-Golgi Membranes and the Intermediate Compartment Localize to Induced Membranes with Distinct Replication Functions in Flavivirus-Infected Cells

Replication of the flavivirus Kunjin virus is associated with virus-induced membrane structures within the cytoplasm of infected cells; these membranes appear as packets of vesicles associated with the sites of viral RNA synthesis and as convoluted membranes (CM) and paracrystalline arrays (PC) containing the components of the virus-specified protease (E. G. Westaway, J. M. Mackenzie, M. T. Kenney, M. K. Jones, and A. A. Khromykh, J. Virol. 71:6650-6661, 1997). To determine the cellular origins of these membrane structures, we compared the immunolabelling patterns of several cell markers in relation to these sites by immunofluorescence and immunoelectron microscopy. A marker for the trans-Golgi membranes and the trans-Golgi network, 1,4-galactosyltransferase (GalT), was redistributed to large foci in the cytoplasm of Kunjin virus-infected cells, partially coincident with immunofluorescent foci associated with the putative sites of viral RNA synthesis. As determined by immunoelectron microscopy, the induced vesicle packets contained GalT, whereas the CM and PC contained a specific protein marker for the intermediate compartment (ERGIC53). A further indicator of the role of cellular organelles in their biogenesis was the observation that the Golgi apparatus-disrupting agent brefeldin A prevented further development of immunofluorescent foci of induced membranes if added before the end of the latent period but that once formed, these membrane foci were resistant to brefeldin A dispersion. Reticulum membranes emanating from the induced CM and PC were also labelled with the rough endoplasmic reticulum marker anti-protein disulfide isomerase and were obviously redistributed during infection. This is the first report identifying trans-Golgi membranes and the intermediate compartment as the apparent sources of the flavivirus-induced membranes involved in events of replication.     

Description de Falklandia gen.n. de I'Ocean Austral et définition des Lysianassoidea uristidiens (Crustacea, Amphipoda)

The revision of the antarctic–subantarctic species Orchomenopsis reducta Schellenberg, 1931, has led to its attribution to a new, highly apomorphic genus: Falklandia gen.n. A new definition of the uristid group is given and Falklandia with 36 other lysianassoid genera are attributed to this supposedly monophyletic group.     

COMPARISON OF ISOZYMES AMONG TYPHA SPECIES IN THE EASTERN UNITED STATES

Biochemical phenotsypes of four taxa of Typha from the eastern United States were determined by starch gel electrophoresis. The isozyme banding patterns of T. latifolia, T. angustifolia and T. domingensis are distinct and allow unambiguous species identification when morphological characters are inadequate or unsuitable. The fourth form, T. glauca, is not an F₁ hybrid, but it does appear to be intermediate between T. latifolia and T. angustifolia. The status of T. glauca and evolutionary relationships among the four forms may now be clarified by additional sampling because of the distinct and relatively invariant isozyme banding patterns which are described.     

Cyanide-Resistant Root Respiration and Tap Root Formation in Two Subspecies of Hypochaeris radicata

Root respiration of the tap root forming species Hypochaeris radicata L. was measured during tap root formation. A comparison was made of two subspecies: H. radicata L. ssp. radicata L., a subspecies from relatively rich soils, and H. radicata L. ssp. ericetorum Van Soest, a subspecies from poor acidic soils. Root respiration was high and to a large extent inhibited by hydroxamic acid (SHAM) before the start of the tap root formation, indicating a high activity of an alternative non-phosphorylative electron transport chain. The rate of root respiration was much lower and less sensitive to SHAM when a considerable tap root was present. However, root respiration was also cyanide-resistant when a tap root was present, indicating that the alternative pathway was still present. A decreased rate of root respiration coincided with an increase of the content of storage carbohydrates, mainly in the tap root. The level of reducing sugars was constant throughout the experimental period, and it was concluded that the activity of the alternative oxidative pathway was significant in oxidation of sugars that could not be utilized for purposes like energy production, the formation of intermediates for growth or for storage. Root respiration decreased after the formation of a tap root. This decrease could neither be attributed to a gradual disappearance of the alternative chain, nor to a decreased level of reducing sugars. No differences in respiratory metabolism between the two subspecies have been observed, suggesting that a high activity of the alternative oxidative pathway is not significant in adaptation of the present two subspecies to relatively nutrient-rich or poor soils.     

Intermediate sucrose octa-acetate sensitivity suggests a third allele at mouse bitter taste locus Soa and Soa-Rua identity

Mice have been characterized as either tasters or non-tastersof the bitter compound sucrose octa-acetate(SOA). However, 11of 17 supposedly non-taster inbred strains were found to avoid1 mM SOA. All 17 strains were indifferent to 0.1 mM SOA. Tasterstrains avoided both concentrations. The intermediate phenotypewas dubbed demitaster. A consistent phenotypic dominance orderwas found in crosses among both inbred and outbred strains (taster> non-taster > demitaster). Demitasters were found (withtasters) in an outbred strain showing monogenic segregationfor SOA avoidance. This, plus monogenic segregation in a back-crossof taster to demitaster inbred strains, suggested a third alleleat the Soa locus (Soa^c). Demitaster allelism was supported bythe strong associations found in 15 strains between the threeSOA phenotypes and HindIII restriction fragment patterns forthe closely linked Prp (proline rich protein) loci. SOA demitasterstrains were also intermediate in raffinose undeca-acetate (RUA)avoidance. Furthermore, B6.SW-Soa² congenic mice avoided notonly SOA, but RUA and eight other acetylated sugars. A previouslyproposed separate RUA-sensitivity gene (Rua) thus appeared tobe redundant.     

Intraspinal Tumors in Children

Thirty cases of intraspinal neoplasms occurring during the first two decades of life are reviewed. Histologic examination showed 13 of these to be astrocytomas, 6 neuroblastomas, 5 sarcomas, 3 ependymomas, 2 neurofibromas and 1 a schwannoma. Orthopedic deformities developed or worsened in 60 percent of patients surviving longer than a year after diagnosis. In five patients some form of endocrine deficiency developed after irradiation. For treatment of radiosensitive extradural malignant lesions, biopsy followed by irradiation is advocated.     

RGD Surface Functionalization of the Hydrophilic Acrylic Intraocular Lens Material to Control Posterior Capsular Opacification

Posterior Capsular Opacification (PCO) is the capsule fibrosis developed on implanted IntraOcular Lens (IOL) by the de-differentiation of Lens Epithelial Cells (LECs) undergoing Epithelial Mesenchymal Transition (EMT). Literature has shown that the incidence of PCO is multifactorial including the patient''s age or disease, surgical technique, and IOL design and material. Reports comparing hydrophilic and hydrophobic acrylic IOLs have shown that the former has more severe PCO. On the other hand, we have previously demonstrated that the adhesion of LECs is favored on hydrophobic compared to hydrophilic materials. By combining these two facts and contemporary knowledge in PCO development via the EMT pathway, we propose a biomimetically inspired strategy to promote LEC adhesion without de-differentiation to reduce the risk of PCO development. By surface grafting of a cell adhesion molecule (RGD peptide) onto the conventional hydrophilic acrylic IOL material, the surface-functionalized IOL can be used to reconstitute a capsule-LEC-IOL sandwich structure, which has been considered to prevent PCO formation in literature. Our results show that the innovative biomaterial improves LEC adhesion, while also exhibiting similar optical (light transmittance, optical bench) and mechanical (haptic compression force, IOL injection force) properties compared to the starting material. In addition, compared to the hydrophobic IOL material, our bioactive biomaterial exhibits similar abilities in LEC adhesion, morphology maintenance, and EMT biomarker expression, which is the crucial pathway to induce PCO. The in vitro assays suggest that this biomaterial has the potential to reduce the risk factor of PCO development.     

Transmission characteristics of Spiroplasma citri and its effect on leafhopper vectors from the Circulifer tenellus complex

Several leafhopper variants of the Circulifer tenellus complex were collected in “citrus stubborn” affected areas in Israel. Two of these variants transmitted the Spiroplasma citri to Matthiola incana after being injected with the disease agent. The variant from Atriplex halimus was designated Circulifer tenellus-A (CTA) and the variant from Portulaca oleracea was designated Circulifer tenellus-? (CTP). Transmission characteristics were determined for both leafhoppers. A high rate of transmission (43.3%) was obtained by single CTA leafhoppers that were injected with the Amiad S. citri isolate from the Upper Galilee, compared with 7% transmission obtained with the CTP leafhoppers. The Gilgal S. citri isolate from the Jordan Valley, was not transmitted by either. Injection was more effective than acquisition access feeding to render the leafhopper infective for both CTA and CTP. The minimum acquisition access period needed for the CTA variant to transmit the Amiad isolate was 1 h. Longer AAPs did not necessarily result in a higher rate of transmission. The minimum incubation period was 6 days and the maximum was 32 days. The LP₅₀ calculated from the logarithmic curve y = 45.74Ln(x)–53.68 was 9.64 days. The minimum inoculation access period (IAP) was lh. The same transmission parameters for the CTP variant could not be determined, as no transmission was obtained even when groups of five-six insects were placed on a single plant.