Clinical and Molecular Characterization of Patients with Distal 11q Deletions

Jacobsen syndrome is caused by segmental aneusomy for the distal end of the long arm of chromosome 11. Typical features include mild to moderate psychomotor retardation, trigonocephaly, facial dysmorphism, cardiac defects, and thrombocytopenia, though none of these features are invariably present. To define the critical regions responsible for these abnormalities, we studied 17 individuals with de novo terminal deletions of 11q. The patients were characterized in a loss-of-heterozygosity analysis using polymorphic dinucleotide repeats. The breakpoints in the complete two-generation families were localized with an average resolution of 3.9 cM. Eight patients with the largest deletions extending from 11q23.3 to 11qter have breakpoints, between D11S924 and D11S1341. This cytogenetic region accounts for the majority of 11q⁻ patients and may be related to the FRA11B fragile site in 11q23.3. One patient with a small terminal deletion distal to D11S1351 had facial dysmorphism, cardiac defects, and thrombocytopenia, suggesting that the genes responsible for these features may lie distal to D11S1351. Twelve of 15 patients with deletion breakpoints as far distal as D11S1345 had trigonocephaly, while patients with deletions distal to D11S912 did not, suggesting that, if hemizygosity for a single gene is responsible for this dysmorphic feature, the gene may lie distal to D11S1345 and proximal to D11S912.     

Fatty Acid Methyl Ester Profiles for Characterization of Glomalean Fungi and Their Endomycorrhizae

Arbuscule-forming fungi in the order Glomales form obligate endomycorrhizal associations with plants that make them difficult to quantify, and taxonomy of the group is only beginning to be objectively understood. Fatty acid methyl ester (FAME) profiles were analyzed to assess the diversity and quantity of fatty acids in 53 isolates of 24 glomalean species. Spores and endomycorrhizal roots of sudan grass (Sorghum sudanense) and the citrus rootstock Carrizo citrange (Poncirus trifoliata x Citrus sinensis) were examined. Spores yielded reproducible FAME profiles from replicate spore collections extracted from soil pot cultures despite being grown in association with a host plant and with contaminating microorganisms present. Unweighted pair group analysis revealed relatively tight clusters of groups at the intraspecific, specific, and generic levels; however, lipid profiles at the family level were convergent. Thus, FAME profile comparisons provided a robust measure of similarity below the family level. FAME profiles in sudan grass roots containing vesicles and/or spores of Glomus intraradices were more similar to spore profiles than to profiles from nonmycorrhizal roots. The FAME profiles for Gigaspora species, which do not form vesicles or spores in roots, were less distinct from nonmycorrhizal roots. G. intraradices and G. rosea produced fatty acids in roots that were distinguishable from each other as well as from the host root. Production in citrus roots of the fatty acid 16:1(inf(omega)5) cis by two Glomus species was correlated with the development of mycorrhizal colonization as measured by clearing and staining procedures and by estimates of total incidence and vesicle intensity. FAME analysis of roots not only provided a measure of colonization development but also served as an index of carbon allocated to intraradical fungal growth and lipid storage.     

Non-invasive monitoring of ovarian function in several felid species by measurement of fecal estradiol-17β and progestins

An extraction and assay procedure to measure fecal estradiol-17β and progestin concentrations in several cat species was developed and validated for use for noninvasive monitoring of ovarian function. Fecal samples were collected over a range of 3–20 months from female tigers (three), lions (three), snow leopards (three), cheetahs (two), caracals (two), and domestic cats (five). Samples were extracted with 90% methanol, lipids removed with petroleum ether, and the estradiol and progestins in the methanol measured by radioimmunoassay (RIA). High Performance Liquid Chromatography (HPLC) fractionation and subsequent RIA of the fractions indicated that the estradiol-17β antiserum cross-reacted primarily with estradiol-17β in the feces of lions and tigers and was assumed to be specific for estradiol-17β in the feces of other species as well. However, there were several immunoreactive compounds, presumably progesterone metabolites, excreted in the feces which varied both quantitatively and qualitatively among species. The behavior of tigers, lions, cheetahs, and caracals was visually monitored during the collection period and frequency of sexual behaviors was positively correlated with increases in fecal estradiol in all species observed. The mean fecal estradiol-17β peaks were as follows: tigers, 128.0 ± 13.1; lions, 186.0 ± 14.8; snow leopards, 136.7 ± 15.9; cheetahs, 140.9 ± 9.0; caracals, 24.5 ± 4.0; and domestic cats 158.9 ± 19.3 ng/gm. Fecal progestin concentrations rose significantly (P < 0.001) only after breeding or during pregnancy and were as follows: tigers, 5.6 ± 0.6; lions, 1.9 ± 0.1; cheetahs, 8.4 ± 1.1; and caracals, 2.4 ± 0.4 μg/gm. Fecal progestins were elevated for one-half to two-thirds of the gestation length during presumed pseudopregnancy but remained elevated throughout successful pregnancies. These results suggest that ovarian function can be monitored noninvasively in the family Felidae by the measurement of fecal estradiol-17β and progestin concentrations. © 1995 Wiley-Liss, Inc.     

Therapy of bovine ocular squamous-cell carcinoma with local doses of interleukin-2: 67% complete regressions after 20 months of follow-up

We have tested the therapeutic potency of peritumorally injected low doses of interleukin-2(IL-2). Seventy tumours of the bovine ocular squamous-cell carcinoma (BOSCC), 1–3 cm in diameter, were treated with 5000, 20 000 or 200 000 U IL-2 from Eurocetus (Chiron) to find the optimal dose for treatment. Injections were given peritumorally on Monday to Friday on 2 consecutive weeks. The size of the tumours was measured before treatment and 1, 3, 4, 9 and 20 months after treatment. After 9 months complete regression was observed in 89% of the tumours treated with 5000 U IL-2, 80% treated with 20 000 U and 67% treated with 200 000 U. After 20 months, there was complete regression of 35%, 31% and 67% of the tumours respectively. The 9-and 20-month results of the 200 000-U treatment are significantly better than those of the 5000-U and 20 000-U treatments taken together. This protocol may be useful to treat advanced inoperable tumours (e.g. of the nasopharynx or skin) of human patients.     

A new monoclonal antibody (3D3) generated with human respiratory mucins and directed against Lewis determinants

We have prepared a monoclonal antibody (MAb), 3D3, raised againstpurified human respiratory mucins. This antibody recognizedmucins and proteolytically derived glycopeptides. The epitoperecognized by the antibody was destroyed by -L-fucosidase, indicatingthat it was present on the carbohydrate moieties. Structuralspecificity was determined by adsorption on a variety of synthetic,insolubilized oligosaccharides. Several lines of evidence indicatethat the 3D3 MAb reacted strongly with the Lewis (Le^b) antigen,but also recognized Le^a and Le^y determinants. This antibodymight be useful to study mucin secretion. human bronchial mucins Lewis b     

Soft rot Erwinia bacteria in surface and underground waters in southern Scotland and in Colorado, United States

An anaerobic liquid enrichment method followed by plating on a selective medium revealed that the soft rot coliform bacterium Erwinia carotovora subsp. carotovora was generally present in water from drains, ditches, streams, rivers and lakes (including reservoirs) in southern Scotland and in Colorado, United States, in mountainous, upland and arable areas through the year. Many sites were remote from susceptible or diseased crops. Erwinia carotovora subsp. atroseptica was isolated much less frequently and no Erwinia bacteria were isolated from underground waters. Erwinia bacteria were also found in rain-water in Scotland, in winter snow from mountain passes in Colorado, and in sea water from the west and east coasts of Scotland and from the coasts of Oregon, California, Texas, Louisiana and Florida. The significance of the occurrence of these bacteria in water is discussed in relation to the control of blackleg and soft rot diseases of potato by production of Erwinia -free stocks.     

Pro-enkephalin gene derived peptides in the porcine stomach: cellular distribution and molecular forms

Antisera to the enkephalin variants Met-enk Arg6Phe7 and Met-enk Arg6Gly7Leu8 have been used in immunohistochemistry and radioimmunoassay studies of hog stomach. In the mucosa of the antrum, but not fundus, there was identified a population of immunoreactive endocrine-like cells. When extracts of antral mucosa were fractionated by gel filtration on Sephadex G50 the predominant immunoreactive forms of Met-enk Arg6Phe7 and Met-enk Arg6Gly7Leu8 were found to elute before the standards, and were compatible with N-terminally extended variants. In the muscle layers of both antrum and fundus, immunoreactive nerve fibers were found, these were especially numerous in the myenteric plexus. In extracts of the antral muscle, 50-60% of both Met-enk Arg6Phe7 and Met-enk Arg6Gly7Leu8 immunoreactivity eluted in the position of the standards and the remainder had the properties of N-terminally extended variants. In the fundus muscle the variants accounted for 70-80% of total activity. The results indicate that the proenkephalin gene is expressed in neurones and endocrine cells of the hog stomach. The different patterns of molecular forms found in different regions of the stomach suggest that the precursor is processed by different pathways in different populations of endocrine cells and neurones.     

Unusual redox behaviour of cytochrome b-561 from bovine chromaffin granule membranes

(1) Redox titrations of cytochrome b-561 have been performed with the purified cytochrome and with intact and detergent-solubilized chromaffin-granule membranes. (2) The midpoint redox potential of the cytochrome is 100–130 mV; this depends upon the composition of the buffer, but is independent of pH in the range 5.5–7.5; partial proteolysis of the cytochrome raises the midpoint potential to 160 mV. (3) The Nernst plots of titration data have slopes of 75–115 mV, and are in some cases sigmoid in shape. This may be explained by negative cooperativity during redox transitions in oligomeric cytochrome b-561. (4) Measurements of the haem and cytochrome content of chromaffin granule membrane suggest a haem content of 1 mol/mol protein. (5) Chemical crosslinking of cytochrome b-561 suggests that it may exist as an oligomer of 4–6 polypeptide chains within the chromaffin granule membrane. Aggregation of purified cytochrome b-561 was shown by gel filtration studies and by immunological methods in SDS-polyacrylamide gels. Studies of the molecular weight of the aggregates suggest that the monomer has a molecular weight close to 22 000, but migrates anomalously slowly during electrophoresis.     

Characterisation of ionisations that influence the redox potential of mitochondrial cytochrome c and photosynthetic bacterial cytochromes c2

Several cytochromes c₂ from the Rhodospirillaceae show a pH dependence of redox potential in the physiological pH range which can be described by equations involving an ionisation in the oxidised form (pK_o) and one in the reduced form (pK_r). These cytochromes fall into one of two groups according to the degree of separation of pK_o and pK_r. In group A, represented here by the Rhodomicrobium vannielii cytochrome c₂, the separation is approx. one pH unit and the ionisation is that of a haem propionic acid. Members of this group are unique among both cytochromes c₂ and mitochondrial cytochromes c in lacking the conserved residue Arg-38. We propose that the role of Arg-38 is to lower the pK of the nearby propionic acid, so that it lies out of the physiological pH range. Substitution of this residue by an uncharged amino acid leads to a raised pK for the propionic acid. In group B, represented here by Rhodopseudomonas viridis cytochrome c₂, the separation between pK_o and pK_r is approx. 0.4 pH unit and the ionisable group is a histidine at position 39. This was established by NMR spectroscopy and confirmed by chemical modification. Only a few other members of the cytochrome c₂/mitochondrial cytochrome c family have a histidine at this position and of these, both Crithidia cytochrome c-557 and yeast cytochrome c were found to have a pH-dependent redox potential similar to that of Rps. viridis cytochrome c₂. Using Coulomb's law, it was found that the energy required to separate pK_o and pK_r could be accounted for by simple electrostatic interactions between the haem iron and the ionisable group.     

Host related factors affecting oviposition behavior in Brachymeria intermedia

Some behavioral and physical defenses of Lymantria dispar (L.) pupae are described. It was found that the layer of webbing surrounding pupae significantly reduced oviposition rates in the pupal parasitoid Brachymeria intermedia (Nees) (Hymenoptera, Chalcididae). The reasons for this reduction and consequent parasitoid responses were investigated. The role of these behaviors in this host parasitoid relationship are discussed.

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Résumé Le comportement défensif des chrysalides de Lymantria dispar L. consiste en cambrage et en tournoiement. Les chrysalides encoconées étaient plus sensibles aux stimuli tactiles que celles qui ont été extraites des cocons. Brachymeria intermedia Nees avait moint de succès dans l'insertion complète de la tarière dans des chrysalides encoconées qu'extraites, car elles devenaient empêtrées dans le cocon quand la chrysalide se défendait. Il y avait différentes réponses du parasitoïde à l'empêtrement. Elles comprenaient l'abandon de l'attaque par un départ, la remise en selle sur l'hôte, la morsure à travers les fils du cocon, l'attaque d'un hôte voisin, le repos ou la toilette. Les taux d'insertion de la tarière pouvaient être augmentés par l'élimination artificielle de quelques fils. La durée des opérations était plus courte sur les chrysalides plus petites. La discussion a porté sur l'importance de ces comportements sur les relations de ces parasitoïdes avec leurs hôtes.