Novel dolichyl derivatives in rat spleen

Novel dolichyl derivatives were found in rat spleen. The compounds were eluted from reverse phase HPLC after eluting dolichyl fatty acid ester. The elution profiles of the unsaponified forms of the unknown compounds were coincident with that of dolichol from spleen on reverse phase HPLC. The compounds were not dolichyl dolichoate, which are present in bovine thyroid. The compounds were not found in young rats (4 months of age) but were found in old rats (above 12 months of age), and they were not detected in other tissues under our conditions.     

槐树组织培养中再生植株的比较解剖学研究

本文对组织培养过程中,槐树(Sophora japonica L.)再生植株正常苗和玻璃苗的叶、茎及茎端的解剖结构进行了比较研究。结果表明:正常苗结构基本类似于实生苗,玻璃苗结构变异较大;玻璃苗叶片变厚,表皮细胞形状不规则,气孔保卫细胞萎缩变形,叶肉无明显的栅栏组织与海绵组织分化,叶绿体含量较少,叶维管组织发育不良;茎横切面形状不规则,表皮上气孔数目较多,皮层厚角组织不明显,维营束大致分布成一轮,形成不规则维管柱;茎端分生组织细胞层数较少,不呈现典型的原套原体结构。     

蓝狐消化道内分泌细胞的免疫组化研究

蓝狐又名北极狐(Alopex lagopus),属于食肉目(Carnivora),犬科(Canidae),北极狐属(Alopex).     

Ingestion of Neospora caninum tissue cysts by Mustela species

Dogs are a definitive host of Neospora caninum, a protozoal parasite that causes abortion in cattle. Mustelids were tested to determine if they could also be definitive hosts. The procedures used were the same as those previously used to test dogs. Ermine (Mustela erminea), weasels (Mustela frenata) and ferrets (Mustela putorius) were fed N. caninum-infected mice. Neospora caninum oocysts were not observed. Mustelid faeces were fed to mice. The mice did not seroconvert and N. caninum was not detected in murine brains using tissue culture and PCR. The hypothesis that Mustela spp. are definitive hosts of N. caninum is not supported.     

An electron microscopic observation of conidium and hypha of Erysiphe graminis hordei

Summary The fine structure of conidia and hyphae ofErysiphe graminis hordei, attacking leaves of barley, were investigated. The cell walls of conidia and hyphae were relatively thin and consisted of two layers, the inner and outer layers. The surface of conidia was not smooth and the thickness of cell walls was irregular. A nucleus, mitochondria, endoplasmic reticula and vacuoles in plasma were identified. The vacuoles in conidia were tightly packed with fine granules. Such granules in vacuoles, however, were not observed in hyphal cells.A lamellar structure was located in conidia, but not in hyphal cells. This structure may be specific in conidia of this fungus, but its function is not yet known. Many glycogen granules were observed in endoplasm of conidia, which were scattered or congregated in groups. In hyphae, however, they were extremely few. Hyphal septa were connected directly with the inner layer of cell walls. These had simple septal pore. The Woronin bodies were detected in the endoplasm in the vicinity of hyphal septa.Contribution No. 192.     

Characterization of murine decidual natural killer (NK) cells and their relevance to the success of pregnancy

The identification of lytic cells in 6.5-day to 9.5-day murine decidua as NK cells has been extended. The cells with natural killer (NK) activity in early decidua were nonphagocytic and heterogeneous in size as assessed by velocity sedimentation at unit gravity. The numbers of lytic cells were reduced by treatment with anti-asialo GM1 in vivo and they were absent from the decidua of bg/bg mice. Thus, decidual NK cells were not distinct from NK cells in other tissues. The decline in the levels of decidual NK activity as pregnancy progressed was attributed to their regulation by other cells present in decidua by midgestation. The development of NK activity in decidua was dependent upon the presence of an embryo, however, decidual NK cells were not essential for successful pregnancy because viable offspring were obtained from mice lacking decidual NK activity. It was shown that NK cells from either spleen or decidua were unlikely to cause damage to embryos during the first half of pregnancy as freshly dissociated 9.5- and 11.5-day embryonic cells resisted NK lysis. Furthermore, blastocysts were not damaged by coincubation with splenic or decidual NK cells and were viable upon subsequent embryo transfer. These studies indicate that decidual NK cells are not essential for successful pregnancy and are not necessarily detrimental to early embryos. It is suggested that decidual NK cells may play other nonimmunological roles during embryonic development.     

Phylogeny of Ephemeroptera (mayflies) based on molecular evidence

This study represents the first molecular phylogeny for the Order Ephemeroptera. The analyses included 31 of the 37 families, representing approximately 24% of the genera. Fifteen families were supported as being monophyletic, five families were supported as nonmonophyletic, and 11 families were only represented by one species, and monophyly was not testable. The suborders Furcatergalia and Carapacea were supported as monophyletic while Setisura and Pisciforma were not supported as monophyletic. The superfamilies Ephemerelloidea and Caenoidea were supported as monophyletic while Baetoidea, Siphlonuroidea, Ephemeroidea, and Heptagenioidea were not. Baetidae was recovered as sister to the remaining clades. The mayfly gill to wing origin hypothesis was not supported nor refuted by these data. Mandibular tusks were supported as having at least one loss in Behningiidae and, together with the burrowing lifestyle, possibly two origins. The fishlike body form was supported as plesiomorphic for mayflies with multiple secondary losses. Topological sensitivity analysis was used as a tool to examine patterns concerning the stability of relationships across a parameter landscape, providing additional information that may not have been acquired otherwise.     

Comparative study of the distribution of semisynthetic cephalosporins in the body of rats]

Distribution of 6 cephalosporin antibiotics, i. e. cephaloridin, cephalotin, cephradin cephacetryl, cephazolin and cephapyrin for parenteral use was studied comparatively on rats. The studies showed that all the above cephalosporins were well absorbed into the blood after intramuscular administration. The highest serum levels were achieved with the use of cephozolin. Still, its levels in the animal organs were mainly not higher and sometimes even lower than those provided by the other antibiotics. The highest levels of cephalosporins were detected in the kidneys. Cephalotin, cephapyrin and cephacetryl differed by the character of their distribution in the rats from the other 3 antibiotics: the levels of cephalotin and cephapyrin in the heart, spleen and muscles were lower than those of the other cephalosporins; sometimes they were even not detected in these organs; cephacetryl was not found in these organs. The levels of these 3 antibiotics in the kidneys were lower than those of the other cephalosporins. Cephalotin, cephacetryl and sometimes cephapyrin were not detected in the rat liver. None of the cephalosporins was found in the brain tissue.     

Inhibition of human natural killer activity by monolayers of primary cell cultures

Some primary and continuous cell cultures were tested for their capacity to regulate human natural killer (NK) activity. Primary cultures of endothelial cells, fetal fibroblasts, adult fibroblasts, amnion epithelial cells, renal parenchymal cells, and ovarian carcinoma cells inhibited NK activity when peripheral blood lymphocytes were preincubated on target cell monolayers for 18 h before testing the cytotoxicity against K-562. The supernatants of the inhibiting cell cultures were not suppressive. Prostaglandins or suppressive lymphocytes were not involved in the phenomenon. The binding capacity of the effector cells was not changed, suggesting that the suppressive signal was targeted at the cytolytic machinery of NK cells. The down-regulating capacity of the cell cultures weakened significantly during subculturing in vitro, and continuous cell lines were not inhibitory. The inactivation of NK cells may be one of the mechanisms by which target cells are protected from NK activity.     

Diminished antigen processing by endosomal acidification mutant antigen-presenting cells

The role of acidified endosomes in Ag processing was investigated using mutant Chinese hamster ovary cells that express temperature sensitive defects in their acidification mechanism. These cells were transfected with MHC class II genes to convert them to APC. When such mutant cells were incubated at the nonpermissive temperature, losing early endosomal but not lysosomal acidification, their ability to process several native protein Ag was impaired. The nonpermissive temperature did not affect Ag processing by transfected wild type parental cells. Furthermore, T cells were stimulated normally under these conditions when the mutant cells presented antigenic peptide fragments, which do not require processing. The mutant cells were also not defective in the uptake and overall degradation of native antigen. The elimination of Ag processing by paraformaldehyde and chloroquine treatment of the Chinese hamster ovary cells indicates that these cells do not use a different process than physiologic APC. These results suggest that acidification of early endosomes is an important event in Ag processing.     

A 30-HZ Pulsed Magnetic Field Can Stop Early Embryonic Development

Chick embryos were exposed in vivo to a 30-Hz magnetic field (MF); the frequency was chosen to produce near-resonance conditions for calcium and sodium ions in the local geomagnetic field. After a 48-hour incubation and exposure to the MF, a morphological analysis of the embryos was performed and they were classified as normal or not normal. The not normals were developed embryos with slight (abnormals) or strong (malformed) abnormalities and non-developed organisms. The results showed that the proportions of normal and not-normal embryos were not modified by the MF. However, significant changes were observed among the not normal embryos: The proportion of abnormals and malformed was decreased, while the non-developed embryos significantly increased. The results support the hypothesis that a resonance-like effect on calcium and/or sodium ions was produced in organisms which already had some developmental defects.     

An actin-like component in spermatocytes of a crane fly (Nephrotoma suturalis Loew)

Decorated actin-like filaments were seen in spindles after crane fly spermatocytes were glycerinated and then treated with rabbit skeletal muscle heavy meromyosin (HMM). Both ATP and pyrophosphate inhibited the HMM reaction. In prometaphase, metaphase, and mid-anaphase cells, actin-like filaments were seen near regions where chromosomal spindle fibres are seen in living cells, and were oriented in the pole-to-pole direction. In the interzone of anaphase cells, actin-like filaments were not oriented in a preferential direction when they were not associated with the microtubules attached to the sex chromosomes. No filaments were seen in glycerinated spindles not treated with HMM. We discuss reasons why filaments might not be seen without prior HMM treatment, and we discuss the possible role of the actin-like filaments in the spindles. — Spindle microtubules often were not seen in cells treated with HMM. This depended on the stage of division: in prometaphase no microtubules were seen; in metaphase microtubules were seen, in apparently normal numbers; in mid-anaphase, microtubules between the autosomes and the poles were seen in reduced numbers, those associated with the equatorial sex-chromosomes were seen in apparently normal numbers, while those between the separating autosomal half-bivalents were not seen. Microtubules were not seen in glycerinated spindles not treated with HMM, suggesting that HMM in some way affects microtubule stability. The question of microtubule stability is briefly discussed.     

Occurrence of Kudoa sp. (Myxozoa) in Trachurus trachurus L. (osteichthyes) in Portugal

Specimens of Trachurus trachurus L., obtained monthly in a fish market of Oporto from October 1998 to August 1999, were examined for the presence of Kudoa in the muscle. Kudoa sp. spores were found in 84.7% of the specimens (n = 209). The prevalence was not significantly different between seasons and was not related to the host's length. The infection does not seem to have negative effects on the fish quality once no macroscopic pseudocysts or myoliquefaction were detected.     

Effect of the carbohydrate growth substrate on polysaccharolytic enzyme formation by anaerobic fungi isolated from the foregut and hindgut of nonruminant herbivores and the forestomach of ruminants

The effect of the carbohydrate growth substrate on polysaccharide-degrading enzyme formation by anaerobic fungi was examined using four strains of Piromyces isolated from hindgut fermenters, three Piromyces isolates from the pre-peptic forestomach of macropodid marsupials, and two ruminal isolates of Neocallimastix spp. The range of enzymes formed by the nine isolates was similar although, under the growth conditions examined, one hindgut isolate did not form amylolytic enzymes. The cellulolytic and xylanolytic enzyme profiles were the same: inter-strain differences in the levels of enzymic activity were apparent, but they were not related to either the genus or intestinal origin of the isolates. Pectin degrading enzymes were not detected in any of the isolates. The cellulolytic and xylanolytic enzymes were formed constitutively during growth on mono-, di- and polysaccharidic carbohydrates but the specific activities were both strain-and substrate-dependent. The activities were considerably lower in glucose-grown preparations of three of the fungi (one each from the hindgut, foregut and rumen) indicating that enzyme synthesis was repressed by glucose; enzyme formation by the other isolates studied was not controlled by catabolite regulatory mechanisms.     

Presence of abnormal synaptonemal complexes in heterothallic species of Neurospora

Synaptonemal complex abnormalities are frequent in reconstructed meiotic prophase nuclei of Neurospora crassa and Neurospora intermedia. Three kinds of synaptonemal complex anomalies were seen: lateral component splits, lateral component junctions, and multiple complexes. The anomalies apparently are formed during or after the pairing process, as they were not seen in the largely unpaired early zygotene chromosomes. Their presence at all the other substages from mid-zygotene to late pachytene indicates that they are not eliminated before the synaptonemal complex decomposes at diplotene. Abnormal synaptonemal complexes were seen in all 19 crosses of N. crassa and N. intermedia that were examined, including matings between standard laboratory strains, inversions, Spore killers, and strains collected from nature. The frequency of affected nuclei and degree of abnormality within a nucleus varied in different matings. No abnormalities were present in the homothallic species Neurospora africana and Neurospora terricola. Structural chromosome aberrations, introgression, and heterozygosity have been eliminated as causes for pairing disorder. The abnormal synaptonemal complexes seemingly do not interfere with normal ascus development and ascospore formation. The affected nuclei are not aborted during meiotic prophase, nor are they eliminated by abortion of mature asci. The abnormal meiocytes do not lead to aneuploidy, as judged by the low frequency of white ascospores in crosses between wild type strains that have many abnormalities. Thus, the abnormal synatonemal complexes do not appear to prevent chiasma formation between homologues.     

Alterations in the carboxy-termianl half of cloacin destabilize the protein and prevent its export by Escherichia coli

Several overlapping carboxy-terminal and internal deletions were constructed in the cloacin structural gene. The expression, the binding of the cloacin DF13 immunity protein and the release into the culture medium of the mutant cloacin polypeptides were studied by immunoblotting and ELISAs. Minor alterations at the carboxy-terminal end of the cloacin did not affect protein expression, stability or release to a large extent, but larger carboxy-terminal deletions strongly destabilized the protein and no release was observed. The removal of a particular region within the carboxy-terminal portion of cloacin strongly destabilized the polypeptide and made it a target for proteolytic degradation. Binding of immunity protein did not affect stability and release of the mutant polypeptides. By using immunoelectron microscopy, the polypeptides that were not exported were located in the cytoplasm of producing cells. Large aggregates of these mutant polypeptides were not observed in the cytoplasm: the polypeptides were present in a soluble form.     

Characterization of the outermembrane proteins of Vibrio cholerae expressed in in vivo culture

Two outer membrane proteins (Omps) of Vibrio cholerae O1, expressed in the intestine (in vivo) but not in culture media (in vitro), were investigated. The molecular masses of those proteins were 116 kDa and 15 kDa, and they were not associated with iron-regulated proteins. Convalescent cholera patients' sera reacted with the 15 kDa protein but not with the 116 kDa protein. The N-terminal amino acid sequence of the 15 kDa protein was homologous to V. cholerae OmpT. Anti-serum to the 15 kDa protein caused agglutination of the organisms grown in the intestine, but not the organisms in in vitro culture. The anti-serum was bactericidal, but it did not inhibit the adhesion of the organisms to the intestine and HEp-2 cells. These findings suggest the possibility that the 15 kDa protein could be involved in post-infection immunity.     

A proteomics approach to the identification of mammalian mitochondrial small subunit ribosomal proteins

Mammalian mitochondrial small subunit ribosomal proteins were separated by two-dimensional polyacrylamide gel electrophoresis. The proteins in six individual spots were subjected to in-gel tryptic digestion. Peptides were separated by capillary liquid chromatography, and the sequences of selected peptides were obtained by electrospray tandem mass spectrometry. The peptide sequences obtained were used to screen human expressed sequence tag data bases, and complete consensus cDNAs were assembled. Mammalian mitochondrial small subunit ribosomal proteins from six different classes of ribosomal proteins were identified. Only two of these proteins have significant sequence similarities to ribosomal proteins from prokaryotes. These proteins correspond to Escherichia coli S10 and S14. Homologs of two human mitochondrial proteins not found in prokaryotes were observed in the genomes of Drosophila melanogaster and Caenorhabditis elegans. A homolog of one of these proteins was observed in D. melanogaster but not in C. elegans, while a homolog of the other was present in C. elegans but not in D. melanogaster. A homolog of one of the ribosomal proteins not found in prokaryotes was tentatively identified in the yeast genome. This latter protein is the first reported example of a ribosomal protein that is shared by mitochondrial ribosomes from lower and higher eukaryotes that does not have a homolog in prokaryotes.     

Acrosomal status and motility of guinea pig spermatozoa during in vitro penetration of the cumulus oophorus

Previous studies have suggested that both acrosome-intact and acrosome-reacted guinea pig sperm are capable of binding to the zona pellucida of cumulus-free oocytes, but the acrosomal status of guinea pig sperm during penetration of the cumulus has not been reported. We made video recordings of the interaction between capacitated guinea pig sperm and cumulus-invested guinea pig oocytes. The videotapes were analysed to identify sperm with hyperactivated motility and to classify the acrosomal status of sperm during penetration of the cumulus and after binding to the zona pellucida. The resolution of the video recordings was not sufficient to recognise sperm with swollen acrosomes. However, sperm that had completed the acrosome reaction were easily identified. Acrosome-reacted sperm were found adherent to the outer boundary of the cumulus, but were never observed to penetrate the cumulus. The percentage of acrosome-intact, hyperactivated sperm was higher in the cumulus oophorus than in culture medium, suggesting that changes in motility were elicited in response to contact with the cumulus. Fully acrosome-reacted sperm were found adherent to the zona pellucida, and solubilised guinea pig zona pellucida was capable of inducing acrosome reactions in capacitated guinea pig sperm. Acrosome-intact sperm were also observed on the zona, but they were not tightly bound and did not have hyperactivated motility, suggesting that these sperm were not functionally capacitated. Our observations demonstrate that guinea pig sperm penetrate the cumulus matrix in an acrosome-intact state. Although we did not observe sperm undergoing the acrosome reaction, our observations and experimental data suggest that the acrosome reaction of guinea pig sperm is completed on or near the surface of the zona pellucida.     

Fine needle aspirates of follicular lesions of the thyroid gland. The intermediate-type smear

In order to evaluate fine needle aspirates of thyroid lesions with features intermediate between those of follicular neoplasms and colloid nodules, 38 aspirates in which a definitive diagnosis had not been made were reviewed. On review, ten aspirates were excluded from the "intermediate" category; seven were reclassified as unsatisfactory and one as a cellular colloid nodule. Two papillary carcinomas showed a complex pattern not identified in smears from other lesions; these aspirates were also classified separately for independent evaluation. The remaining 28 aspirates were characterized by syncytial-type tissue fragments with mild nuclear atypia. The association of syncytial-type tissue fragments and orderly sheets and fragments forming a honeycomb pattern in the same aspirate indicated a colloid nodule, though a two-disease process could not be excluded. Of the aspirates containing only syncytial-type tissue fragments, 50% were from adenomas, 25% were from carcinomas, and 25% were from colloid nodules. Criteria to distinguish between the various follicular lesions were not identified in these smears.