Effect of processing soybeans on their use by calves

Changes in the fine structure of the hyphae were studied in the course of spore formation by Micropolyspora fascifera. Sporulating hyphae differ from vegetative hyphae by a less dense cytoplasm, a large zone of the nucleoid with distinctly fibrillar structure, and a thin layerless cell wall. Spore formation is accompanied with autolysis of the vegetative hyphae, which consists in vacuolization of the hyphae and appearance of a large number of tubular membrane structures within them. Spores are formed, like in Actinomyces spp., by simultaneous division of the hyphae with septa; the structure of sporulating septa is similar to that of the fragmenting mycelium in Nocardia; the structure of mature spores is similar to that in some Actinomyces spp. Therefore, M. fascifera differs from other Micropolyspora spp. not only by the chemical composition of the cells (the presence of nocardiomicolic acids) but also by their structure. Taxonomic position of the species is discussed.     

HIV risk behaviors: a comparison of U.S. Hispanic and Dominican Republic youth

Using 3-week-old male beagle dogs, we examined the cartilage-bone replacement processes in the mandibular condyle by means of light and electron microscopy. Calcification of the cartilage matrix occurred in the central area of the longitudinal septa, but not in the transverse septa. Perivascular mononuclear cells absorbed the transverse septa which initiated the opening of the chondrocytic lacunae. These cells phagocytosed septal cartilage fragments. Shortly thereafter, a thin bone layer was deposited on the remaining longitudinal septa by invading osteoblasts. Osteoclasts in lacunae developed neither ruffled borders nor clear zones in the cartilage matrix, but once the bone layer has been deposited in the remaining cartilage, these structures formed. Our results suggest that the cartilage-bone replacement in mandibular condyle involves three sequential processes: 1) degradation and phagocytosis of cartilage fragments in the transverse septa by mononuclear cells, 2) bone deposition over the remaining longitudinal septa, and 3) degradation of both calcified cartilage and bone by osteoclasts.     

Loss of growth polarity and mislocalization of septa in a Neurospora mutant altered in the regulatory subunit of cAMP-dependent protein kinase

In filamentous fungi, growth polarity (i.e. hyphal extension) and formation of septa require polarized deposition of new cell wall material. To explore this process, we analyzed a conditional Neurospora crassa mutant, mcb, which showed a complete loss of growth polarity when incubated at the restrictive temperature. Cloning and DNA sequence analysis of the mcb gene revealed that it encodes a regulatory subunit of cAMP-dependent protein kinase (PKA). Unexpectedly, the mcb mutant still formed septa when grown at the restrictive temperature, indicating that polarized deposition of wall material during septation is a process that is, at least in part, independent of polarized deposition during hyphal tip extension. However, septa formed in the mcb mutant growing at the restrictive temperature are mislocalized. Both polarized growth and septation are actin-dependent processes, and a concentration of actin patches is observed at growing hyphal tips and sites where septa are being formed. In the mcb mutant growing at the restrictive temperature, actin patches are uniformly distributed over the cell cortex; however, actin patches are still concentrated at sites of septation. Our results suggest that the PKA pathway regulates hyphal growth polarity, possibly through organizing actin patches at the cell cortex.     

Septate uterus: detection and prediction of obstetrical complications by different forms of ultrasonography

The aims of the study were to compare the accuracy of transvaginal ultrasonography, transvaginal color Doppler sonography, hysterosonography, and three-dimensional ultrasonography in detection of septate uterus and to evaluate the occurrence of obstetrical complications in relation to septal dimension and vascularity. Each patient underwent transvaginal ultrasonography and color Doppler examination, whereas hysterosonography and three-dimensional ultrasonography were carried out in 76 and 86 patients, respectively. The sensitivity of different sonographic imaging modalities ranges between 95.21% and 99.29%. Color and pulsed Doppler sonographic studies of the septal area revealed vascularity in 71.22%. Patients with vascularized septa had a higher prevalence of obstetrical complications than those with avascularized septa (P < 0.05). Three-dimensional ultrasonography and hysterosonography are highly accurate diagnostic tools for detection of uterine septa. We found no correlation between septal dimension and rate of obstetrical complications, although pregnancy loss was most likely to occur in patients with vascularized septa.     

Autoactivation of spore germination in mutant and wild type strains of Dictyostelium discoideum

Freshly formed wild type Dictyostelium discoideum spores are constitutively dormant, and thus require an activation treatment to germinate. Wild type spores may germinate without an activation treatment (autoactivate) after a period of ageing (maturation) in the intact fruiting body. Mutants have been isolated which autoactivate without the need for ageing. Autoactivation of mutant and aged wild type spores appears to occur by identical mechanisms; thus the mutation may involve premature maturation. Autoactivation is mediated by autoactivator substances released from spores as they spontaneously swell. These factors are readily chromatographed, and elute from a Biogel P2 column in three peaks of activity. One activity peak appears only after spores have begun to germinate. No autoactivator substances are released from heat activated spores. Autoactivation is sensitive to cychloheximide, and may result from altered spore permeability. Autoactivation is likely to be the mechanism of D. discoideum spore germination in nature.     

Effect of volatile inhibitors from natural and amended soils on germination of sclerotia of Macrophomina phaseolina

Studies on volatile soil fungistasis showed that sclerotia of Macrophomina phaseolina are not sensitive to volatiles liberated by natural soils with varying pH values. Soil amended with alfalfa hay (1%, w/w) liberated volatiles fungistatic to sclerotia only when the assay was performed 2 weeks or less after amendment. Chitin-amended soils liberated fungistatic volatiles from 1 to 3 weeks after amendment. Indirect evidence indicated that ammonia may be the active volatile in amended soil. It was concluded that fungistatic volatiles may not play an important role in the survival of sclerotia in nature.     

Structural roles of the spore coat proteins in Dictyostelium discoideum

The integrity of spores formed by mutant strains of Dictyostelium discoideum lacking the major spore coat proteins, SP96, SP70, or SP60, was compared to that of wild-type strains. Single, double, and triple knock-out strains developed normally and produced spores which were indistinguishable from wild-type spores by light or electron microscopy. However, the mutant strains were susceptable to staining with the lectin, ricin A, which recognizes a galactose-rich polysaccharide that is normally hidden by overlying spore coat proteins. The intensity of staining with fluorescently labeled ricinA increased as the spore coat proteins were incrementally lost. While these results indicate that the major outer spore coat proteins are not essential for the construction of a multi-layered spore coat in Dictyostelium, they show that the spores are more porous which might make them at risk to predators before germination.     

Sporicidal action of peracetic acid and protective effects of transition metal ions

Although peracetic acid (PAA) is used widely for cold sterilization and disinfection, its mechanisms of sporicidal action are poorly understood. PAA at high concentrations (5-10%) can cause major loss of optical absorbance and microscopically-visible damage to bacterial spores. Spores killed by lower levels of PAA (0.02-0.05%) showed no visible damage and remained refractile. Treatment of spores of Bacillus megaterium ATCC 19213 with PAA at concentrations close to the lethal level sensitized the cells to subsequent heat killing. In addition, PAA was found to act in concert with hypochlorite and iodine to kill spores. Antioxidant sulfhydryl compounds or ascorbate protected spores against PAA killing. Trolox, a water-soluble form of alpha-tocopherol, was somewhat protective, while other antioxidants, including alpha-tocopherol, urate, bilirubin, ampicillin and ethanol were not protective. Chelators, including dipicolinate, were not protective, but transition metal ions, especially the reduced forms (Co2+, Cu+ and Fe2+) were highly protective. The net conclusions are that organic radicals formed from PAA are sporicidal and that they may act as reducing agents for spores that are normally in a highly oxidized state, in addition to their well known actions as oxidizing agents in causing damage to vegetative cells.     

Ultrastructural investigation of the spore-forming protist Nephridiophaga blattellae in the Malpighian tubules of the German cockroach Blattella germanica

Multinuclear plasmodia of the sporogenic protist Nephridiophaga blattellae are found intracellularly and in the lumen of the Malpighian tubules of the German cockroach Blattella germanica. Spore formation occurs only in the lumen. During sporogony, about 10-35 spores measuring 5.5x3.2 microm are endogenously formed within a plasmodium. Sporoblasts arise by the fusion of cisternae of the endoplasmic reticulum into a double membranous wall, which encloses a generative nucleus plus a portion of cytoplasm. Several somatic nuclei remain in the residual cytoplasm. Sporoblast and residual cytoplasm include mitochondria of the tubular type, endoplasmic reticulum, and many free ribosomes. During spore maturation, electron-dense wall material is deposited between the spore membranes, and the spores gain their typical oval, biconcave form. Freeze-etched spores reveal a small, central, cap-like structure, which may be the site where an infectious sporoplasm could emerge. Mature spores always have one nucleus, whereas early sporoblast stages with two small nuclei were found by transmission electron microscopy. Many nuclei of different developmental stages contain granules within the nuclear envelope. The systematic position of N. blattellae is unresolved. In certain respects it is reminiscent of Haplosporidia. However, the organisms of the two groups have different spore-forming processes and haplosporosomes are missing in the nephridiophagids. Therefore a new phylum might have to be erected for members of the family Nephridiophagidae.     

Impact of Particle Aggregated Microbes on UV Disinfection. I: Evaluation of Spore–Clay Aggregates and Suspended Spores

Aggregation of microbes with particles can reduce the effectiveness of ultraviolet (UV) disinfection. This study evaluated the comparative impact of dispersed spores, dispersed spores mixed with clay particles (nonaggregated), spore–spore aggregates, and spore–clay aggregates on UV disinfection performance in simulated drinking waters. Aggregates were induced by flocculation with alum and characterized by particle size analysis (count, volume, and surface area) of dispersed and aggregated systems, scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) analysis. It was concluded that spores within aggregates of the spore–clay system were protected from UV irradiation compared to nonaggregated spores and the difference between these systems was found to be statistically significant throughout the UV range tested. In addition SEM-EDX analysis suggested that aggregate composition is nonhomogeneous with respect to the ratio of spores and clay particles among aggregates. It was estimated that 30–50% of the spores in the aggregates tested were protected from UV irradiation.     

Isokotanins A-C: new bicoumarins from the sclerotia of Aspergillus alliaceus

Three new bicoumarin metabolites, isokotanins A [1], B [2], and C [3], were isolated from the sclerotia of Aspergillus alliaceus. Isokotanin A is a regioisomer of the known bicoumarin kotanin [4]. The structures and spectral assignments for 1-3 were determined on the basis of selective INEPT, HMQC, and NOESY nmr data, as well as by chemical interconversions. Isokotanins B and C show activity against the corn earworm Helicoverpa zea and the dried fruit bettle Carpophilus hemipterus. The known compounds kotanin [4], desmethylkotanin [5], nominine, and paspaline were also isolated from extracts of A. alliaceus sclerotia.     

Efficacy of mint and eucalyptus leaves on the physicochemical characteristics of stored wheat against insect infestation

Variation in Alternaria spores in the atmosphere of Murcia (SE Spain) were studied for 2 years. Using a Burkard volumetric sampler located about 19 m above ground level, spores were collected and counted hourly by light microscopy. The results suggest that species of Alternaria are present in the atmosphere of Murcia every day throughout the year. Mean daily concentration of Alternaria was 25 spores/m3. Seasonal variations showed the lowest concentrations during winter and the highest in autumn. In the monthly pattern two peaks were observed every year: the first in about May-June and the second occurring in October. In the intradiurnal variations, maximum concentrations were observed between 5:00-6:00 p.m. (Spanish official time). The variation of Alternaria spores in the atmosphere in Murcia seems to be related to different factors throughout the year.     

Performance characteristics of a whole-body PET scanner

METHODS: This study characterizes the performance of a newly developed whole-body PET scanner (Advance, General Electric Medical Systems, Milwaukee, WI). The scanner consists of 12,096 bismuth germinate crystals (4.0 mm transaxial by 8.1 mm axial by 30 mm radial) in 18 rings, giving 35 two-dimensional image planes through an axial field of view of 15.2 cm. The rings are separated by retractable tungsten septa. Intrinsic spatial resolution, scatter fraction, sensitivity, high count rate performance and image quality are evaluated. RESULTS: Transaxial resolution (in FWHM) is 3.8 mm at the center and increases to 5.0 mm tangential and 7.3 mm radial at R = 20 cm. Average axial resolution decreases from 4.0 mm FWHM at the center to 6.6 mm at R = 20 cm. Scatter fraction is 9.4% and 10.2% for direct and cross slices, respectively. With septa out, the average scatter fraction is 34%. Total system sensitivity for true events (in kcps/(microCi/cc)) is 223 with septa in and 1200 with septa out. Dead-time losses of 50% correspond to a radioactivity concentration of 4.9 (0.81) microCi/cc and a true event count rate of 489 (480) kcps with septa in (out). Noise-equivalent count rate (NECR) for the system as a whole shows a maximum of 261 (159) kcps at a radioactivity concentration of 4.1 (0.65) microCi/cc with septa in (out). NECR is insensitive to changes in lower gamma-energy discrimination between 250-350 keV. CONCLUSIONS: The results show the performance of the newly designed PET scanner to be well suited for clinical and research applications.     

Reproduction of nuclei in Pelomyxa palustris

Light and electron micrographs were made of nuclei in Pelomyxa palustris, a unicellular, multinucleated giant amoeboid organism. We analyzed 1019 pelomyxae and classified their nuclei according to their location in the nuclear cycle. The majority of organisms (56.3%) had interphase nuclei, some of which contained spores of mostly 1-3 microns in diameter. The nuclei had disintegrated in 1.3% of organisms that appeared to have no nuclei. The remainder (42.4%) had nuclei in the form of spores (1 to 10 microns spheroids) that were in various stages of development and growth. Mitotic figures were seen in some of them, with several chromosome pairs per nucleus. Interchromosomal fibers were seen at anaphase, and newly formed "young" interphase nuclei were observed.     

Metabolism of L-threonine and its relationship to sclerotium formation in Sclerotium rolfsii

The activities of L-threonine dehydrogenase (I), 2-amino-3-oxybutyrate:CoA ligase (II), malate synthetase (III), isocitrate lyase (IV), glyoxylate dehydrogenase (V), glycine decarboxylase (VI), L-serine hydroxymethyltransferase (VII), glucan synthetase (VIII), glucose 6-phosphate dehydrogenase (IX) and succinic dehydrogenase (X) were detected in cell-free extracts prepared from the mycelium of the fungus Sclerotium rolfsii type R. Transfer of S. rolfsii to a threonine-containing medium resulted in a significant increase in the intracellular concentrations of L-threonine, glycine, serine and glyoxylate, and a decrease in oxalate. Incubation with 14C-labelled L-threonine resulted in an immediate output of 14CO2, and an accumulation of labelled glycine and serine in the mycelium. L-Threonine (10(-2)M) increased branching, favoured formation of sclerotia, and induced the formation of enzymes I to VIII, but not IX and X. Sodium oxalate (1-5 X 10(-2)M) inhibited branching, sclerotium formation and the activity of enzymes III and IV. Glycine (10(-1) M) inhibited branching, sclerotium formation and activity of I and II. Ammonium chloride (10(-1) to 10(-2) M) inhibited formation of sclerotia, threonine uptake and activity of III. Acetyl-CoA inhibited V and L-cysteine inhibited I as well as sclerotium formation and branching. It is suggested that hyphal morphogenesis and formation of sclerotia in S. rolfsii require an increased supply of carbohydrate intermediates and energy and that these are mainly supplied by the glyoxylate pathway.     

32P-post-labelling analysis of DNA adducts formed in the upper gastrointestinal tissue of mice fed bracken extract or bracken spores

Bracken toxicity to both domestic and laboratory animals is well established and tumours are formed when rodents are treated with either bracken extracts or bracken spores. In this study we have administered bracken spores and extract to mice in order to investigate whether such exposure leads to the formation of DNA adducts. DNA, isolated from the upper gastrointestinal tract and liver, was digested to 3'-nucleotides. Adducts were extracted with butanol, 32P-post-labelled, separated by thin layer chromatography (TLC) and visualised and quantified using storage-phosphor technology. A cluster of adducts was clearly seen in the DNA of the upper gastrointestinal tract, but not liver, 5 and 24 h after treatment with bracken extract or bracken spores. These adducts were not observed in DNA extracted from vehicle-treated animals. Whereas, after 5 h adduct levels in extract and spore-treated animals were similar, after 24 h adduct levels in the extract-treated animals had diminished by > 75%, but levels in spore-treated animals remained similar to those found after 5 h. This suggests that the DNA-reactive compounds were being released slowly from the spores, even though the spores had been sonicated before administration. Adducts were also quantified after the addition of an internal standard (deoxyinosine 3'-monophosphate) by comparing the amount of label incorporated into the adducts with that found in a known amount of the internal standard. Adduct levels using this internal standard approach were similar to those found by direct measurement of radioactivity incorporated into the adduct, indicating that the labelling of adducts was quantitative. We have tried, unsuccessfully, to synthesise ptaquiloside, the principal carcinogenic component present within bracken. However, similar patterns of adducts were observed when two other compounds, (1-(4-chlorophenyl sulphonyl)-l-cyclopropane carbonitrile and 3-cyclopropylindeno [1,2-c] pyrazol-4-(O-methyl)oxime), which both contain a cyclopropyl ring, were administered to mice. The adducts detected in bracken-treated animals may, thus, have arisen from ptaquiloside but, whether these adducts arise directly from the compounds and bracken spores/extract themselves or via an indirect mechanism, remains to be determined. As bracken-induced DNA adducts are detectable in rodent tissues by a 32P-post-labelling procedure commonly employed to investigate DNA damage in human populations, it may prove possible to apply such approaches to determine human exposure.     

Structural changes induced by glycine on Streptomyces antibioticus

Germination and vegetative growth of Streptomyces antibioticus in liquid medium with different concentrations of glycine was examined. Both processes proved to be sensitive to the amino acid, being inhibited by 5 and 2.5% glycine, respectively. At concentrations of 5% or more, lysis of the vegetative mycelium occurred. Subinhibitory concentrations of glycine induced structural changes on germinating spores. These included an increase in the number of germ tubes produced by spore, in relation to the control. Moreover, soon after outgrowth the tubes bifurcate, giving rise to germinated spores with a characteristic aspect, and anomalous formation of cross-walls that appear both within the spores and in the newly formed germinative tubes, at or close to the region of outgrowth. The branching effect of glycine was also observed during vegetative growth of S. antibioticus.     

The Kota of the Nilgiri Hills: a demographic study

We report the existence of a surface configuration in a true spore-forming genus having cell wall constituents of type IV. Micropolyspora angiospora, M. caesia and M. faeni freeze-fractured along the wall surface and only had a surface configuration on their aerial mycelium and aerial spores, with none on the substrate mycelium and substrate spore. The surface configuration of Micropolyspora was distinctly more complicated than that of Nocardia. The aerial spores of M. angiospora were characteristic in that they possessed ridges, two kinds of surface configurations (i.e. rodlets and fibres), and a complex pattern on the surface. Some rodlets of this organism were formed of a two-stranded helix, each strand having a diameter similar to that of a fibre.     

Microarchitecture of the cat testis with special reference to Leydig cells: a three-dimensional study by alkali maceration method and scanning electron microscopy

Testes from adult cats were studied by means of parallel transmission and scanning electron microscopy (SEM) after NaOH digestion technique, which selectively removed connective tissues or cells. The testis is covered by a dense fibroconnective tunica albuginea that partially divides the organ in lobules by sending septa into the parenchyma. The lamina propria of the seminiferous tubules consisted of one or two rows of cells. The interstitium was made up of randomly arranged collagen bundles. The most significant feature was the numerous Leydig cells rich in lipid droplets and displaying epithelioid features. Following alkali digestion and SEM these cells showed a cord-like arrangement. The cords were formed by one or two closely apposed cells, in between which some labyrinthine or canalicular-like spaces were left that in some areas opened in wide perivascular spaces. This particular arrangement of Leydig cells and the labyrinthine intercellular spaces is very likely designed to improve cell secretion of hormones, facilitating their transport into the blood, as well as the traffic of fluids and metabolites. The present techniques allowed the visualization of a real three-dimensional testicular microarchitecture and microtopography, not detectable with other methods. Such a study may help to better highlight the testicular morphophysiology.