Acyl-homoserine lactones suppresses IEC-6 cell proliferation and increase permeability of isolated rat colon

We investigated to determine whether a variety of acyl-homoserine lactones (AHLs) influences epithelial cell proliferation and mucosal permeability. 3-Oxo-C₁₂-homoserine lactone (HSL) and 3-oxo-C₁₄-HSL significantly suppressed IEC-6 cell proliferation. A significant increase in mucosal permeability was observed in isolated rat colon tissue exposed to C₁₂-HSL, 3-oxo-C₁₂-HSL, and 3-oxo-C₁₄-HSL. These data indicate that AHLs suppress epithelial proliferation and disrupt barrier function in intestinal mucosa.     

Effects of germinated barley foodstuff on microflora and short chain fatty acid production in dextran sulfate sodium-induced colitis in rats

Germinated barley foodstuff (GBF) administration has been previously reported to suppress dextran sulfate sodium (DSS)-induced experimental colitis. In this study, we investigated the roles of the intestinal microflora and short chain fatty acids (SCFAs) following administration of GBF in DSS-induced rat colitis. Sprague-Dawley rats were fed 3% (w/w of diet) DSS in GBF-diets for 5 days. The control rats were fed 3% DSS in cellulose-diets for 5 days. The administration of GBF effectively prevented bloody diarrhea and mucosal damage as compared to control rats. GBF significantly elevated fecal acetic acid and n-butyric acid levels. GBF tended to increase the number of eubacteria and that of bifidobacteria as compared to control rats. In addition, the number of enterobacteriaceae, the total number of aerobes and bacteroidaseae, were significantly lower in rats fed GBF than in the control group. It is suggested that the therapeutic effects of GBF for DSS-induced colitis depend mainly on increased SCFAs, which are accompanied by changes of composition of intestinal bacteria.     

Host-related variability in arbuscular mycorrhizal fungal structures in roots of <Emphasis Type="Italic">Hedera rhombea</Emphasis>, <Emphasis Type="Italic">Rubus parvifolius</Emphasis>, and <Emphasis Type="Italic">Rosa multiflora</Emphasis> under controlled conditions

The arbuscular mycorrhizal (AM) morphology of three host plant species inoculated with single and mixed fungal culture and the distribution of AM fungal species in roots of the hosts treated with a mixed culture of AM fungi were determined. The aim was to investigate the effect of host plants and AM fungi on AM morphology of coexisting plant species. Noncolonized rooted cuttings of Hedera rhombea (Miq) Bean, Rubus parvifolius L., and Rosa multiflora Thunb. were inoculated with five fungal species as single and mixed culture inocula. The fungal species used were Gigaspora rosea and Scutellospora erythropa, previously isolated from H. rhombea; Acaulospora longula and Glomus etunicatum from R. parvifolius; and Glomus claroideum from both plant species. A few hyphal and arbusculate coils were seen in the mixed culture-inoculated roots of R. parvifolius; all fungal treatments produced this Paris-type AM in H. rhombea and Arum-type AM in R. parvifolius, and R. multiflora indicates that AM morphology is strongly controlled by the identity of the host plants used in this study. AM fungal rDNA was extracted separately from roots of each replicate plant species inoculated with the mixed fungal culture, amplified, cloned, sequenced, and analyzed to determine the AM fungal species and their respective proportions in roots of each plant species. Glomus etunicatum and G. claroideum of the family Glomaceae generally occurred more frequently in R. parvifolius and R. multiflora, which form Arum-types, whereas S. erythropa, of the family Gigasporaceae, was the most frequently detected species in H. rhombea, which produced Paris-type AM. Although the genotype of the plant species used appears to determine the AM morphologies formed, there was preferential association between the hosts and AM fungal inoculants.     

Restoration of Ca2+-transport in sarcoplasmic reticulum ATPase solubilized with octaethyleneglycol mono n-dodecyl ether

One mg protein/ml of sarcoplasmic reticulum (SR) membranes isolated from rabbit skeletal muscle were solubilized with 50 mg/ml of octaethyleneglycol mono n-dodecyl ether (C12E8) in a solution containing 5 mM CaCl2, 0.1 M KCl, and 20% glycerol at pH 7.5. When 30 mg/ml of soybean lecithin was added to this mixture and then incubated with Bio-beads SM-2 at 20 degrees C for 1.5 h to remove the detergent from the mixture, proteoliposomes were formed. This process restored Ca2+-uptake activity to approximately 50% of that of control sR. However, Ca2+-transport was not observed when SR membranes were formed without the addition of soybean lecithin. The reconstituted vesicles also catalyze Ca2+-release, which is coupled to the backward reaction which forms ATP from ADP and P1 in the presence of a Ca2+-gradient across the membrane. When the reconstituted vesicles were subjected to equilibrium centrifugation in a 5 to 25% glycerol density gradient, all of the Ca2+-transport activity was closely associated with the fraction containing soybean liposome.     

Disease in children infected with HIV in Abidjan,C?te d'Ivoire.

OBJECTIVE--To document the range of disease in African children infected with HIV. DESIGN--Necropsy results in consecutive children aged 1 month or more who were HIV positive and in children who were HIV negative for comparison; IgA western blots on serum samples from children under 2 years of age who were positive for HIV-1 to test the validity of routine HIV serology. SETTING--Largest hospital in Abidjan, Côte d''Ivoire. SUBJECTS--78 children who were HIV positive and 77 children who were HIV negative on whom a necropsy was performed; their median ages at death were 18 and 21 months respectively. 36 HIV positive children and 29 HIV negative children were 1-14 months old; 42 HIV positive and 48 HIV negative children were > or = 15 months old. MAIN OUTCOME MEASURES--Cause of death and prevalence of diseases confirmed pathologically. RESULTS--Respiratory tract infections were more common in HIV positive than in HIV negative children (73 (94%) v 52 (68%); P < 0.05), and were aetiologically heterogeneous. Pneumocystis carinii pneumonia was found in 11 out of 36 (31%) HIV positive children aged < 15 months, but in no HIV negative children. Among older children measles was more common in HIV positive children (8/42 (19%) v 2/48 (4%); P < 0.06). Pyogenic meningitis was present in similar proportions of HIV positive and HIV negative children aged < 15 months (7/36 (19%) and 7/29 (24%)). In HIV positive children tuberculosis (1/78), lymphocytic interstitial pneumonitis (1/78), and HIV encephalitis (2/78) were rare. CONCLUSIONS--There is greater overlap between diseases associated with HIV infection and other common health problems in African children than there is in adults. Compared with adults, HIV positive children had a high prevalence of P carinii pneumonia and a low prevalence of tuberculosis. Measles, but not malaria, was associated with HIV infection.     

Characterization of a camptothecin-resistant human DNA topoisomerase I in an in vitro system for Simian virus 40 DNA replication.

DNA topoisomerase I was required for bidirectional DNA replication in an in vitro system for Simian virus 40 (SV40) DNA replication with purified proteins in which the replication fork moved at the rate of 260 nucleotides/min on average. DNA topoisomerase I purified from camptothecin-resistant human lymphoblastoid cells, which confers high resistance of cellular DNA replication to camptothecin [Andoh, T., Ishii, K., Suzuki, Y., Ikegami, Y., Kusunoki, Y., Takemoto, Y. & Okada, K. (1987) Proc. Natl Acad. Sci. USA 84, 5565-5569], was characterized using this system. The activity of stimulating bidirectional DNA replication was comparable between two topoisomerase I from parental and resistant cells, i.e. in its dose-response relationship and in its time course for DNA synthesis. Camptothecin severely inhibited the leading as well as the lagging strand synthesis in the reaction containing the wild type topoisomerase I but not the mutant type topoisomerase I. The mutant type topoisomerase I was over 125-fold as resistant to camptothecin as the wild type topoisomerase I. These results are in good agreement with those on the sensitivity of cellular DNA synthesis to camptothecin in the resistant cells. These findings suggest that topoisomerase I is involved in cellular DNA replication as a swivelase and the mutation conferring camptothecin-resistance on the enzyme does not affect its functional efficiency in this system.     

Genetic divergence between the sibling species of river-sculpin,Cottus amblystomopsis andC. nozawae,with special reference to speciation

Synopsis Polypatric speciation from an amphidromous ancestor has been suggested for a fluvial gobyRhinogobius flumineus; a similar history may explain life-history and genetic variation inCottus species. We used starch-gel electrophoresis to examine intra- and interspecific genetic differentiation of two siblingCottus species, amphidromousC. amblystomopsis and fluvialC. nozawae, which are presumed to be in an ancestor-descendant phyletic relationship. Genetic differentiation was markedly less withinC. amblystomopsis, than inC. nozawae. One group ofC. nozawae populations was closer toC. amblystomopsis (by Nei's genetic distance) than it was to other populations ofC. nozawae. This suggests that the differentiation of presently identifiable groups ofC. nozawae populations might have occurred polypatrically from an amphidromous ancestral population at different times. The two groups are presumed to be different species in spite of their quite similar morphological and ecological characteristics: we propose to call this type of speciation Polypatric and parallel speciation. This model of Polypatric and parallel speciation might be generalized to explain evolution of amphidromous or anadromous species to fluvial or lacustrine land-locked species.     

Comparison of Genomic and Epigenomic Expression in Monozygotic Twins Discordant for Rett Syndrome

Monozygotic (identical) twins have been widely used in genetic studies to determine the relative contributions of heredity and the environment in human diseases. Discordance in disease manifestation between affected monozygotic twins has been attributed to either environmental factors or different patterns of X chromosome inactivation (XCI). However, recent studies have identified genetic and epigenetic differences between monozygotic twins, thereby challenging the accepted experimental model for distinguishing the effects of nature and nurture. Here, we report the genomic and epigenomic sequences in skin fibroblasts of a discordant monozygotic twin pair with Rett syndrome, an X-linked neurodevelopmental disorder characterized by autistic features, epileptic seizures, gait ataxia and stereotypical hand movements. The twins shared the same de novo mutation in exon 4 of the MECP2 gene (G269AfsX288), which was paternal in origin and occurred during spermatogenesis. The XCI patterns in the twins did not differ in lymphocytes, skin fibroblasts, and hair cells (which originate from ectoderm as does neuronal tissue). No reproducible differences were detected between the twins in single nucleotide polymorphisms (SNPs), insertion-deletion polymorphisms (indels), or copy number variations. Differences in DNA methylation between the twins were detected in fibroblasts in the upstream regions of genes involved in brain function and skeletal tissues such as Mohawk Homeobox (MKX), Brain-type Creatine Kinase (CKB), and FYN Tyrosine Kinase Protooncogene (FYN). The level of methylation in these upstream regions was inversely correlated with the level of gene expression. Thus, differences in DNA methylation patterns likely underlie the discordance in Rett phenotypes between the twins.