Surface waves and crustal structure on Mars.

We detected surface waves from two meteorite impacts on Mars. By measuring group velocity dispersion along the impact-lander path, we obtained a direct constraint on crustal structure away from the InSight lander. The crust north of the equatorial dichotomy had a shear wave velocity of approximately 3.2 kilometers per second in the 5- to 30-kilometer depth range, with little depth variation. This implies a higher crustal density than inferred beneath the lander, suggesting either compositional differences or reduced porosity in the volcanic areas traversed by the surface waves. The lower velocities and the crustal layering observed beneath the landing site down to a 10-kilometer depth are not a global feature. Structural variations revealed by surface waves hold implications for models of the formation and thickness of the martian crust.

Detailed analysis by Fabry-Perot method of slab photonic crystal line-defect waveguides and cavities in aluminium-free material system.

A single-line-defect low-loss photonic crystal waveguide based on a perforated GaAs membrane in an aluminium-free material system is demonstrated. The GaInP lattice is matched to GaAs as the cladding/sacrificial layer. Fabry-Perot resonances are analyzed to obtain the group velocity dispersion for a 1-mm long guide. The losses are deduced to be close to 5 dB/cm, taking into account the wavelength dependent reflectivity of the guide extremities. In this framework, side-coupled nanocavities are also investigated. Feasibility of low-loss photonic-crystal-based devices combined with a reliable industrial material systems is thus demonstrated.

Prevalence of a trypanosomatid in the southern green stink bug, Nezara viridula.

The southern green stink bug, Nezara viridula (L.), and certain of its host plants were examined to determine the prevalence and biological characteristics of an intestinal trypanosomatid. Promastigotes with short (< or = 17.5 microm excluding flagellum) and long forms (> or = 25.0 microm) usually infected < 50% of the bugs before August and > 50% (maximum 95%) during August-October, but prevalence was not host-density dependent. The flagellate was detected in adults and in all nymphal instars, at all sampling sites where at least 10 bugs were captured, and in bugs from all host plants sampled (soybean, red clover, vetch). Of bugs with flagellates, 27% were heavily infected (> 20 flagellates per 160X microscope field). Weights of infected and uninfected adults did not differ. Live flagellates were detected in bug feces and in one stem of red clover. When bugs were fed soybean pods, tomatoes, or snap beans in the laboratory, only once were flagellates detected in plant tissue (snap beans). The flagellate was cultured in modified Medium 199. This flagellate is prevalent in N. viridula populations in Louisiana and apparently does not cause significant pathological effects in N. viridula or its host plants, including soybean.

Stressors and rearing diseases of Trichoplusia ni: evidence of vertical transmission of NPV and CPV.

Inability to consistently rear healthy Trichoplusia ni led to a study of its rearing diseases. Four diseases were designated after preliminary research which included electron microscopy: cytoplasmic polyhedrosis (due to cytoplasmic polyhedrosis virus, or CPV), nuclear polyhedrosis (due to nucleopolyhedrovirus, or NPV), "neonate death" syndrome (mortality in first or second instars), and "late-instar" syndrome (death in late instars accompanied by bacterial decomposition). Infectious agents were not detected by electron microscopy in insects with the latter two diseases. Prevalence of CPV and NPV, but not the neonate-death or late-instar diseases, in progeny was significantly associated with pairs of mating adults. In conjunction with egg-surface decontamination, this indicated that both viruses may be transmitted transovarially. Pupae killed by CPV had virions in the cytoplasm of infected cells, but polyhedra were empty, not occluding virions. None of the diseases had a consistent pattern of prevalence associated with the date on which eggs were laid after oviposition began. Prevalence rates of cytoplasmic polyhedrosis, nuclear polyhedrosis, and the late-instar disease were significantly greater at 95-100% relative humidity (RH) than at RH levels of 75% or below. These same three diseases killed significantly more insects in crowded rearing conditions (four or five larvae per cup with 10.2-cm(2) diet surface) than in uncrowded conditions (one to three larvae per cup). As a result of these experiments, healthy T. ni have been reared for 10 generations by use of a modified Pasteur method and rearing cups containing no more than two larvae.

Mosquito (Diptera: Culicidae) host compatibility and vector competency for the human myositic parasite Trachipleistophora hominis (Phylum Microspora).

Microsporidian spores of Trachipleistophora hominis Hollister, isolated from a human, readily infected larval stages of both Anopheles quadrimaculatus Say sensu lato and Culex quinque-fasciatus Say. Mosquito infections with T. hominis were located, primarily, in abdominal muscles in segment numbers 4 through 6; other spores were found in the hemocoel and proboscis. Nearly 50% of the infected mosquito larvae survived to the adult stage. Spores recovered from adult mosquitoes were inoculated into mice and resulted in significant muscle infection at the site of injection. Preliminary observations also showed that T. hominis spores can be passively transferred from infected mosquitoes to a sugar water substrate.

The plaque matrix (PQM) and tubules at the surface of intramuscular parasite, Trachipleistophora hominis.

Surface plaque matrix (PQM) and a tubular arrangement of filaments border Trachipleistophora hominis parasites during growth within host muscle. The PQM at the parasite surface forms a network of processes which can be associated with filamentous tubules. Peroxidase tracer delineated the PQM and showed apparent connections with the tubules. The tubules at the interface of T. hominis-infected cells are structurally similar to the extrasporular tubules of the microsporidian, Ameson michaelis. The extrasporular tubules of A. michaelis and the proteins from T. hominis-infected muscle reacted to keratin antibodies, K8.13, K4 and K13. Conversely, antibodies produced to T. hominis-infected muscle, reacted with the extrasporular tubular proteins of A. michaelis. The PQM and tubular elements are thought to play an important role in affecting molecular traffic between the host and parasite.

Evidence for heterotrimeric GTP-binding proteins in Toxoplasma gondii.

Toxoplasma gondii, an intracellular protozoan parasite, resides within a host-derived vacuole that is rapidly modified by a parasite-secreted membranous tubular network. In this study we investigated the involvement of heterotrimeric G proteins in the secretory pathway of T. gondii. Aluminum fluoride (AlFn), a specific activator of heterotrimeric G proteins, induced secretion from isolated tachyzoites of T. gondii in vitro, as seen by light optics and electron microscopy. In Western blot analyses, antibodies to G protein alpha subunits reacted with 39-42 kDa proteins from T. gondii isolates. Antibodies to G(o) alpha and Gs alpha coupled to the fluorescent probe fluorescein isothiocyanate localized to the paranuclear region of T. gondii. Gi3 alpha immunoprobes were confined to the cytoplasmic matrix of T. gondii and also labeled the parasitophorous vesicle. Fluorescein isothiocyanate-conjugated GA/1, an antipeptide antisera directed toward the GTP binding site common to G protein alpha subunits, was confined to the lateral cytoplasmic domain of the parasites where secretion is most prominent. In time-sequence studies using the GA/1 probe, the immunoreactive material shifted position during invasion of target cell to areas of active secretion.

Development and ultrastructure of Trachipleistophora hominis n.g., n.sp. after in vitro isolation from an AIDS patient and inoculation into athymic mice.

Continuous culture was achieved in several cell lines of a microsporidium obtained from the skeletal muscle of an AIDS patient. Development in COS-1 and RK13 cells was prolific. Spores from the original biopsy were also inoculated into athymic mice by i.m. and i.p. routes. Infection was found in several organs as well as in skeletal muscle after a few weeks. All stages were surrounded by an electron-dense surface coat. Meronts had 2-4 nuclei and divided by binary fission. In sporogony the surface coat became separated from the plasma membrane to form a sporophorous vesicle, within which division into sporoblasts was effected by repeated binary fissions. The number of sporoblasts (and later spores) within the sporophorous vesicles varied from 2 to > 32 and the sizes of the vesicles varied, according to the number of spores contained therein, from 5 microns diameter to 14.0 x 11.0 microns. Spores measured 4.0 x 2.4 microns and had a prominent posterior vacuole. The parasite differs from the genus Pleistophora in that it does not form multinucleate sporogonial plasmodia and that the sporophorous vesicle enlarges during sporogony and its wall is not a multilayered structure. It is proposed to place it in a new genus and species Trachipleistophora hominis n.g., n.sp.

Survey for blood parasites in redheads (Aythya americana) wintering at the Chandeleur Islands, Louisiana.

We detected no infections with species of Plasmodium, Haemoproteus, Leucocytozoon, Sarcocystis or Trypanosoma in blood smears, liver and spleen impressions, and muscle tissue from 136 redheads (Aythya americana) collected or captured at the Chandeleur Islands, Louisiana (USA), during three winters (1987 to 1990). One bird, a juvenile male, was infected with an unidentified species of microfilaria. Thus, we found no evidence that hematozoa had an effect on redheads during the course of the study.

Protein-Membrane Interaction Is Essential to Normal Assembly of the Microsporidian Spore Invasion Tube.

Changes in the protein-membrane interaction during assembly of the microsporidian spore invasion tubes were followed by electron microscopy, by video imaging with differential interference contrast (DIC), and by the fluorescent probes 4',6-diamidino-2-phenylindole (DAPI) and 9-diethylamino-5H-benzo{alpha}phenoxazine-5-one (Nile red). Microsporidian spore invasion tubes form by the eversion of polar filament protein (PFP) and presumptive extrusion apparatus (EAP) membrane. Both of these components are essential for formation of the invasion tube. The results indicate that the behavior of the EAP membrane is greatly affected by the position and chemical state of the PFP at the eversion area that constitutes the advancing tube terminal assembly site (TAS). Visual evidence indicates that the EAP membrane is the vehicle for PFP and that this membrane also provides the envelope that surrounds the sporoplasm after its passage through the invasion tube.

Overcoating of Toxoplasma parasitophorous vacuoles with host cell vimentin type intermediate filaments.

The interaction between the Toxoplasma parasitophorous vacuole and vimentin-type intermediate filaments in Vero cells was investigated via immunofluorescence microscopy. A significant rearrangement of host cell vimentin around the Toxoplasma parasitophorous vacuoles occurs throughout the course of infection. Host cell vimentin associates with the parasitophorous vacuoles within an hour after invasion. This vimentin overcoating of the vacuole is initiated at the host cell nuclear surface. During parasite multiplication, vimentin retains a closely defined association with the cytosolic surface of the parasitophorous vacuole. In addition, the vimentin intermediate filaments originating from the host cell nuclear surface are progressively rearranged around the enlarging parasitophorous compartment. During infections, the order of vimentin cytoskeleton is normal throughout the cell and appears redefined only at the vicinity of the parasitophorous vacuole. Depolymerization of the intermediate filaments was achieved with the phosphatase inhibitors okadaic acid and calyculin A. Disruption of the intermediate filament networks resulted in displacement of the parasitophorous vacuoles from the host cell nuclear surface. The data indicate that host cell vimentin binds to the Toxoplasma parasitophorous vacuoles and that the host intermediate filament network serves to dock the parasite compartment to the host cell nuclear surface.

Cytokeratin and Desmoplakin Analogues within an Intracellular Parasite.

A significant amount of the total protein in the spore sacs of the microsporidian Thelohania sp. consisted of the cytoskeletal elements, cytokeratin intermediate filaments, and the desmosomal analogues. The cytokeratin and desmosomal analogues were organized as cage envelopes surrounding the spores within the spore sac stage. Thelohania sp. parasitizes the skeletal muscle of Callinectes sapidus, a crustacean that does not appear to have cytokeratins or desmosomes. Immunoprobe data indicate Thelohania sp. has a 240 kDa desmoplakin protein and 48, 51, 54 and 56 kDa cytokeratin polypeptides responsive to antibodies developed against bovine cytoskeletal counterparts. The cytoskeletal envelopes within the Thelohania sp. spore sac stage appear to enhance the stability and viability of the spores.

Reaginic and homocytotropic IgG antibody response of Mastomys natalensis in experimental infections of filarial parasites (Litomosoides carinii, Dipetalonema viteae, Brugia malayi, B. pahangi).

Reaginic and homocytotropic IgG antibodies in sera using passive cutaneous anaphylaxis (PCA) test and antigen from Litomosoides carinii were followed in Mastomys natalensis, infected with L. carinii, Dipetalonema viteae, Brugia malayi or B. pahangi. Groups of animals with infections of various ages so as to cover a total infection period of up to 300 to 420 days post-infection (p.i.), depending on the species of parasites, were bled at 1- to 3-week intervals over periods of 50-112 days. In addition, intradermal tests were performed on animals infected with L. carinii to detect immediate type hypersensitivity. Reaginic antibodies were usually first detected in the 3rd week after infection. Thereafter, a marked increase of PCA titres was observed in the 4th week p.i. leading to maximum titres 4 weeks after infection with D. viteae and B. pahangi and 6 weeks after B. malayi infection. Mean maximum titres were between 1:40 and 1:160. Following the peak response, titres decreased markedly until the beginning of patency in infections with D. viteae, B. malayi and B. pahangi whereas a constant course was observed at this time in animals infected with L. carinii. A further rise in PCA titres occurred in all infections around the beginning of patency, resulting in maximum reagin levels in L. carinii infections (mean titre 1:80) and moderate titres in the other infections. During early patency there was an inverse relationship between microfilaraemia density and levels of reaginic antibodies. However, in the phase of decreasing parasitaemia in L. carinii infected animals, microfilariae counts and PCA titres were directly correlated. Homocytotropic IgG antibodies showed relatively constant PCA titres of about 1:20 in L. carinii infected Mastomys throughout the observation period. In D. viteae infections they were demonstrated at 30 days p.i., reaching titres of about 1:40. B. malayi infected animals showed a maximum titre of 1:40 40 days p.i.. Thereafter, titres decreased continuously and homocytotropic IgG antibodies were absent at 110 days p.i.. High titres were observed at day 150 but thereafter sera were negative. B. pahangi infected animals showed moderate titres (1:5) 35 days p.i.. Thereafter, antibodies were found at low titres until 115 days p.i.. Intradermal reactions in L. carinii infected animals generally increased in size from 30-60 but decreased when microfilariae appeared in the blood.(ABSTRACT TRUNCATED AT 400 WORDS)

Toxoplasma modifies macrophage phagosomes by secretion of a vesicular network rich in surface proteins.

Modification of macrophage phagosomes begins shortly after formation as Toxoplasma cells secrete membranous vesicles that form a reticulate network within the vacuole. The Toxoplasma-modified compartments then resist normal endocytic processing and digestion. We have used the pronounced Ca++-dependent stability of the intraphagosomal membrane (IPM) network to purify and characterize the structural proteins of this assembly. In addition to the structural matrix, Toxoplasma secretes a discrete set of soluble proteins, including a newly described 22-kD calcium-binding protein. The IPM network adheres to intact Toxoplasma cells after host cell lysis in the presence of 1 mM Ca++; however, the network readily disperses in calcium-free buffer and was purified as vesicles that sedimented at 100,000 g. Purified IPM vesicles were specifically recognized by immune sera from mice with chronic Toxoplasma infection and consisted primarily of a 30-kD protein when analyzed by SDS PAGE. IPM network proteins share a major antigenic component located on the surface of extracellular Toxoplasma cells as shown by immunoperoxidase electron microscopy using a polyclonal antibody prepared against the IPM vesicles. Moreover, in Toxoplasma-infected macrophages, anti-IMP antibody confirmed that the extensive IPM array contains proteins also found on the Toxoplasma cell surface. Our results indicate the IMP network represents a unique structural modification of the phagosome comprised in part of Toxoplasma surface proteins.