Epicuticular wax accumulation and fatty acid elongation activities are induced during leaf development of leeks

Epicuticular wax production was evaluated along the length of expanding leek (Allium porrum L.) leaves to gain insight into the regulation of wax production. Leaf segments from the bottom to the top were analyzed for (a) wax composition and load; (b) microsomal fatty acid elongase, plastidial fatty acid synthase, and acyl-acyl carrier protein (ACP) thioesterase activities; and (c) tissue and cellular morphological changes. The level of total wax, which was low at the bottom, increased 23-fold along the length of the leaf, whereas accumulation of the hentriacontan-16-one increased more than 1000-fold. The onset of wax accumulation was not linked to cell elongation but, rather, occurred several centimeters above the leaf base. Peak microsomal fatty acid elongation activity preceded the onset of wax accumulation, and the maximum fatty acid synthase activity was coincident with the onset. The C16:0- and C18:0-ACP-hydrolyzing activities changed relatively little along the leaf, whereas C18:1-ACP-hydrolyzing activity increased slightly prior to the peak elongase activity. Electron micrographic analyses revealed that wax crystal formation was asynchronous among cells in the initial stages of wax deposition, and morphological changes in the cuticle and cell wall preceded the appearance of wax crystals. These studies demonstrated that wax production and microsomal fatty acid elongation activities were induced within a defined and identifiable region of the expanding leek leaf and provide the foundation for future molecular studies.

Expression of constitutive and tissue-specific acyl carrier protein isoforms in Arabidopsis.

We have characterized the occurrence and expression of multiple acyl carrier protein (ACP) isoforms in Arabidopsis thaliana (L.) Heynh ecotype Columbia. Immunoblot analysis of ACPs from Arabidopsis tissues separated by native polyacrylamide gel electrophoresis and 1 molar urea polyacrylamide gel electrophoresis revealed a complex pattern of multiple ACP isoforms. All tissues examined (leaves, roots, and seeds) expressed at least three forms of ACP. The immunoblot identifications of ACP bands were confirmed by acylation of ACP extracts with Escherichia coli acyl-ACP synthetase. A full-length cDNA clone has been isolated that has 70% identity with a previously characterized Arabidopsis genomic ACP clone (ACP-1) (MA Post-Beittenmiller, A Hlousek-Radojcic, JB Ohlrogge [1989] Nucleic Acids Res 17: 1777). Based on RNA blot analysis, the cDNA clone represents an ACP that is expressed in leaves, seeds, and roots. In order to identify the protein products of each known ACP gene, their mature coding sequences have been expressed in E. coli. Using polymerase chain reactions, exons II and III of the genomic ACP-1 clone and the mature coding sequences of the ACP-2 cDNA clone were subcloned into E. coli expression vectors. Site-directed mutagenesis was used to convert the amino acid sequence of the ACP-2 cDNA clone to that of the A2 clone of Lamppa and Jacks ([1991] Plant Mol Biol 16: 469-474), ACP-3. The three E. coli-expressed proteins have different mobilities on polyacrylamide gel electrophoresis gels and each comigrates with a different Arabidopsis ACP isoform expressed in leaves, seeds, and roots. Thus, all of the three cloned ACPs appear to be constitutively expressed Arabidopsis ACPs. In addition to these three ACP isoforms, protein blots indicate that seed, leaf, and root each express one or more tissue-specific isoforms.

Immunoregulatory factors contributing to fetal allograft survival.

A mammalian fetus expresses a variety of antigens potentially unknown to the immunologically competent mother. Presented here are the results of investigations of maternal immune reactivity to paternally derived antigens of fetoplacental unit, detected at various levels: 1) spleen and distant lymphatic organs, 2) regional lymph nodes draining uterus, and 3) materno-fetal interface. The results suggest that the mother's immune system reacts differently in semiallogeneic pregnancies than in syngeneic ones. The type of the systemic immune response depends on the stage of pregnancy. Increased percentage of CD8+ cells and decreased CD4+/CD8+ cell ratio was found in distant and regional lymphatic organs during pregnancy. The paternal class I MHC antigens expressed on the trophoblast cells are nonpolymorphic molecules which can have a role in immunotrophism of the placenta and in fetal allograft protection.

Expression and analysis of the rat placental class I cDNA clone encoding the Pa antigen.

The previously sequenced cDNA clone pARI.5 was recloned into the mammalian expression vector pcEXV3, and transient and permanent transfectants were prepared in COS7 green monkey kidney fibroblasts. The transfectants were analyzed by indirect immunofluorescence using monoclonal and polyclonal antibodies raised in specifically selected rat strain combinations. These studies showed that pARI.5 encodes the Pa antigen and that the Pa molecule is distinct from the Aa molecule. Probes were derived from the pARI.5 clone and used to study the genomic DNA from Pa-positive and Pa-negative strains. Two probes derived from the 3' untranslated region (3'apARI.5 and 3'bpARI.5) and one probe derived from the 5' region (5'pARI.5) hybridized nonspecifically in all strains under moderate stringency conditions. By contrast, an Xba I restriction fragment unique to the Pa gene was detected with the 5'pARI.5 probe under high stringency conditions. This probe hybridized with a 1.8 kilobase (kb) fragment in the Pa-positive strains and with a 1.7 kb band in the Pa-negative strains. These studies suggest that the gene encoding the Pa antigen, or a fragment thereof, is present in both Pa-positive and Pa-negative strains but may not be expressed in the latter.

[Benign mesenchymal tumors of the uterus]. / O benignim mezenhimnim tumorima uterusa.

Benign mesenchymal tumours of the uterus are discussed. The results are compared with those of other authors and respective epidemiological findings are put forward. Clinical and histological characteristics of uterine myomas and adenomyomas are described in more detail. Irregular bleeding is most frequent among clinical symptoms; it is caused by the persistence of the proliferative endometrium. The secretory conversion of the ectopic endometrium in adenomyoma appears in the glandular epithelium only, while the stromal response to progesteron activity seems inadequate. Maturation of the ectopic and the uterine endometrium is discordant. The cause is the retardation of both the proliferative and secretory phases in the ectopic endometrium. After secretory conversion of the ectopic endometrium the exhaustion and destruction of glands appears. The regeneration of the ectopic endometrium fails to occur because of the absence of the basal layer in the ectopic endometrium. Therefore the authors suppose that the regression of the ectopic endometrium in adenomyomas might be the result of ovulatory cycles.

[Evaluation of diagnostic methods in anatomic urinary incontinence in gynecology]. / Valorizacija dijagnostickih metoda kod anatomske inkontinencije urina u ginekologiji

The present study was undertaken with the aim of verifying the value of diagnostic methods in female stress incontinence. In their diagnostic work the authors used urethrocystography, measuring of the urethral length and resistance. Displacement of the internal urinary meatus related to the interior border of the symphysis was also observed. In demonstrable incontinence the average posterior urethrovesical angle was 140 degrees, and in the absence of incontinence 120 degrees. It was also found that the difference between the pressure of the internal urinary meatus in straining and non-straining women with incontinence was +0,7 mm Hg and in those without incontinence +4,5 mm Hg. It has been observed, furthermore, that the relation between the vertical displacement of the internal urinary meatus in rest and stress was less than 5 in women with incontinence. In women without incontinence it was more than 5. There was no difference in the range of the urethral length in women either with or without stress incontinence.