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Background: Organ transplantation was one of the greatest achievements of medical science during the 20th century. Knowledge, education, and culture all play prominent roles in transplantation because of the complexity of the process from donation to transplantation., Purpose: The aim of this research was to determine and analyze the knowledge and attitudes about organ donation and transplantation among the general population in Limassol, Cyprus., Methods: A quantitative research approach was followed, and a questionnaire consisting of closed-ended questions was completed by adults from the general population in Limassol., Results: One thousand two hundred adults out of the 1,346 adults who were contacted responded to the survey (response rate: 89%) and were included as participants. Of the participants, 93.4% (p < .05) considered organ donation to be lifesaving, 57% expressed interest (and 39.8% expressed disinterest) in becoming organ donors, 80.6% (p < .05) expressed awareness of there being a waiting list for people in need of organ transplantation, 50.4% agreed that brain death must be confirmed before organ removal for transplantation, and 47% recalled having been informed about organ donation through the media, with 31.5% stating that they had never been informed about organ donation., Conclusions: The participants demonstrated limited awareness regarding the organ donation system in Cyprus. Furthermore, a significant percentage stated that they lacked a source for obtaining related information. The Cypriot society should be informed and encouraged to participate in organ donation to increase the rate of organ transplantation., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2020 The Authors. Published by Wolters Kluwer Health, Inc.)

Livestock production in the European Union EU is highly dependent on imported soybean, exposing the livestock farming system to risks related to the global trade of soybean. Lupin species could be a realistic sustainable alternative source of protein for animal feeding. Lupinus is a very diverse genus with many species. However, only four of them-namely, L. albus , L. angustifolius , L. luteus and L. mutabilis -are cultivated. Their use in livestock farming systems has many advantages in relation to economic and environmental impact. Generally, lupin grains are characterized by high protein content, while their oil content is relatively low but of high quality. On the other hand, the presence of quinolizidine alkaloids and their specific carbohydrate composition are the main antinutritional factors that prevent their use in animal feeding. This research is mainly related to L. albus and to L. angustifolius , and to a lesser extent, to L. lauteus and L. mutabilis. The breeding efforts are mostly focused on yield stabilization, resistance to biotic and abiotic stresses, biochemical structure associated with seed quality and late maturing. Progress is made in improving lupin with respect to the seed quality, as well as the tolerance to biotic and abiotic stress. It has to be noted that modern cultivars, mostly of L. albus and L. angustifolius , contain low levels of alkaloids. However, for future breeding efforts, the implementation of marker-assisted selection and the available genomic tools is of great importance.

Sodium nitroprusside (SNP) represents one of the most commonly used NO donors in biological sciences, which acts as a signal molecule in plants responsible for the regulation of the expression of many defense-related enzymes. This study attempts to provide novel insight into the effect of application of low (100 μΜ) and high (2.5 mM) concentrations of SNP on antioxidant gene expression (cAPX, GST, FeSOD, CAT, and AOX) in mature (40 day) and senescing (65 day) Medicago truncatula plants. Quantitative real-time RT-PCR suggests that low concentration of SNP applied in mature leaves leads to an overall induction of antioxidant gene expression, while increasing concentration results in suppression of these genes. Conversely, older plants demonstrate a much more variable regulation which appears to be time dependent. The observed transcriptional regulation pattern in mature M. truncatula plants comes in support of the previously documented protective or damaging effect of SNP depending on concentration applied, whereas senescing M. truncatula plants demonstrated a general suppression in antioxidant gene expression levels regardless of SNP concentration, indicative of reduced overall plant defense capacity against free radicals.

Introduction: Detection of coronary artery calcification (CAC) allows for a refined prediction of cardiovascular risk beyond global risk assessment algorithms. Newer-generation, high-resolution, flat-panel digital detector (FPDD) fluoroscopic systems may provide higher CAC detection rates compared with older fluoroscopic devices., Methods: We compared the CAC detection rates of two fluoroscopic techniques in two different cohorts of asymptomatic individuals, analyzed within a two-decade time interval., Results: FPDD detected CAC more frequently than the older fluoroscopy device, in the more recent and the older patient cohort of individuals, respectively. After propensity score matching to account for differences in age and risk factor prevalence, the adjusted rates of CAC detection remained higher in favor of FPDD (37.7% vs. 23.7%, p=0.026)., Conclusions: The ability of newer cine-fluoroscopic systems to identify CAC in a larger number of asymptomatic, intermediate-risk individuals may have implications for further risk stratification, management of risk factors and long-term prognosis.

The prevalence of prasugrel contraindications and specific conditions requiring precaution for its use in a real world acute coronary syndrome (ACS) population is not known. We performed a prospective descriptive study in 1016 consecutive moderate to high risk ACS patients. In 646 patients (63.6%) subjected to percutaneous coronary intervention, analysis of absolute contraindications (history of stroke/transient ischemic attack or active bleeding), relative contraindications and specific conditions (age ≥ 75 years and/or weight < 60 kg) for prasugrel theoretical administration was performed. In 242 (37.5%) patients there was at least one absolute or relative contraindication or specific condition requiring attention for its use. Overall, 23.1% of patients in our cohort had a prior stroke/transient ischemic attack and/or specific condition to be considered for prasugrel administration. Specifically, the prevalence of stroke/TIA was 3.6%, the prevalence of patients ≥75 years 20% and the prevalence of patients weighing <60 kg 2.2%. Among patients ≥75 years old, 63 (9.8%) had diabetes mellitus or previous myocardial infarction, consisting a high risk subgroup that might benefit from prasugrel administration. In a real world ACS population a relatively high proportion of patients have a potential contraindication for prasugrel administration or necessitate special attention for its use.

Objectives: β-Thalassemias are inherited hemolytic anemias with a broad phenotype and an increased rate of vascular complications despite a near absence of coronary artery disease., Methods: We investigated the presence of endothelial dysfunction and subclinical atherosclerosis in non-transfusion-dependent patients with β-thalassemia intermedia (β-TI) by means of flow-mediated (FMD) and flow-independent (FID) brachial artery dilatation and carotid artery intima-media thickness., Results: In 20 nondiabetic young adults with β-TI, FMD (6.6 ± 3.7 vs. 10.3 ± 3.1%; p = 0.002) and FID (14.0 ± 4.7 vs. 18.0 ± 5.6%; p = 0.02) were both lower relative to the values in 20 matched control subjects, whereas the intima-media thickness was increased (0.51 ± 0.09 vs. 0.46 ± 0.07 mm; p = 0.049). Fibrin generation, soluble endothelial activation markers, and proinflammatory proteins were higher in the patient group, while the plasma cholesterol level was lower., Conclusions: These findings indicate premature atherosclerosis among patients with β-TI; this is in accord with the high incidence of noncoronary vascular episodes in β-TI., (Copyright © 2011 S. Karger AG, Basel.)

Despite the remarkable advances in revascularization strategies made during the last decade, a significant proportion of patients are excluded from either percutaneous coronary intervention or coronary artery bypass grafting because of unsuitable coronary anatomy. Diffuse severe coronary artery disease, small vessel caliber, chronic total occlusions, or extremely calcified vessels are frequent reasons for deferring revascularization with either percutaneous coronary intervention or coronary artery bypass grafting. We present a case concerning a middle-aged asymptomatic patient who was treated successfully with percutaneous coronary intervention due to a chronic total occlusion lesion of the left anterior descending artery. Coronary angiography is an inadequate method for the estimation of the burden of atherosclerotic disease in an artery fed by collaterals. Assessment of any residual antegrade flow, and ipsilateral and contralateral collateral filling of the segments distal to the occlusion with invasive or noninvasive techniques, could affect the appropriate decision-making by physicians.

The triterpenes are a large and diverse group of plant natural products that have important functions in plant protection and food quality, and a range of pharmaceutical and other applications. Like sterols, they are synthesised from mevalonate via the isoprenoid pathway, the two pathways diverging after 2,3-oxidosqualene. During triterpene synthesis 2,3-oxidosqualene is cyclised to one of a number of potential products, the most common of these being the pentacyclic triterpene beta-amyrin. Plants often produce complex mixtures of conjugated triterpene glycosides which may be derived from a single triterpene skeleton. The delineation, functional analysis and exploitation of triterpene pathways in plants therefore represent a substantial challenge. Here we have carried out high throughput screening to identify mutants of diploid oat (Avena strigosa) that are blocked in the early steps of triterpene synthesis. We also show that mutants that are affected in the first committed step in synthesis of beta-amyrin-derived triterpenes, and so are unable to cyclise 2,3-oxidosqualene to beta-amyrin (sad1 mutants), accumulate elevated levels of primary sterols. The major differences were in Delta-7-campesterol and Delta-7-avenasterol, which both increased several fold relative to wild-type levels. This is presumably due to accumulation of squalene and 2,3-oxidosqualene and consequent feedback into the sterol pathway, and is consistent with previous reports in which specific oxidosqualene cyclase inhibitors and elicitors of triterpene biosynthesis were shown to have inverse effects on the flux through the sterol and triterpene pathways., (Copyright 2010 Elsevier Ltd. All rights reserved.)

We investigated the uptake of aluminum (Al) and transport to shoots in two inbred maize lines (Zea mays L., VA-22 and A(4/67)) differing in Al tolerance. Seedlings were grown for 7 days in hydroponic culture with nutrient solution that contained 0, 240, 360, and 480microM Al at pH 4.2. After 7 days of exposure to Al, roots of sensitive maize line (A(4/67)) plants accumulated 2-2.5 times more Al than roots of tolerant line (VA-22) plants. Inductively coupled plasma atomic emission spectrometry (ICP-AES) showed that the tolerant line retained higher concentrations of Ca(2+), Mg(2+), and K(+) compared with the sensitive line. In response to Al treatment, proline (Pro) concentration increased three-fold in roots of tolerant plants, while a slight increase was observed in roots of sensitive-line plants. A substantial carbon surplus (two-fold increase) was observed in roots of the Al-tolerant maize line. Carbohydrate concentration remained almost unchanged in roots of Al-sensitive line plants. Al treatment triggered the enhancement of lipid peroxidation in the sensitive line, while no change in lipid peroxidation level was observed in the tolerant maize line. These data provide further support to the hypothesis that a mechanism exists that excludes Al from the roots of the tolerant maize line, as well as an internal mechanism of tolerance that minimizes accumulation of lipid peroxides through a higher Pro and carbohydrate content related to osmoregulation and membrane stabilization.

Avenacins are antimicrobial triterpene glycosides that are produced by oat (Avena) roots. These compounds confer broad-spectrum resistance to soil pathogens. Avenacin A-1, the major avenacin produced by oats, is strongly UV fluorescent and accumulates in root epidermal cells. We previously defined nine loci required for avenacin synthesis, eight of which are clustered. Mutants affected at seven of these (including Saponin-deficient1 [Sad1], the gene for the first committed enzyme in the pathway) have normal root morphology but reduced root fluorescence. In this study, we focus on mutations at the other two loci, Sad3 (also within the gene cluster) and Sad4 (unlinked), which result in stunted root growth, membrane trafficking defects in the root epidermis, and root hair deficiency. While sad3 and sad4 mutants both accumulate the same intermediate, monodeglucosyl avenacin A-1, the effect on avenacin A-1 glucosylation in sad4 mutants is only partial. sad1/sad1 sad3/sad3 and sad1/sad1 sad4/sad4 double mutants have normal root morphology, implying that the accumulation of incompletely glucosylated avenacin A-1 disrupts membrane trafficking and causes degeneration of the epidermis, with consequential effects on root hair formation. Various lines of evidence indicate that these effects are dosage-dependent. The significance of these data for the evolution and maintenance of the avenacin gene cluster is discussed.

Cell expansion is a central process in plant morphogenesis, and the elongation of roots and root hairs is essential for uptake of minerals and water from the soil. Ca2+ influx from the extracellular store is required for (and sets the rates of) cell elongation in roots. Arabidopsis thaliana rhd2 mutants are defective in Ca2+ uptake and consequently cell expansion is compromised--rhd2 mutants have short root hairs and stunted roots. To determine the regulation of Ca2+ acquisition in growing root cells we show here that RHD2 is an NADPH oxidase, a protein that transfers electrons from NADPH to an electron acceptor leading to the formation of reactive oxygen species (ROS). We show that ROS accumulate in growing wild-type (WT) root hairs but their levels are markedly decreased in rhd2 mutants. Blocking the activity of the NADPH oxidase with diphenylene iodonium (DPI) inhibits ROS formation and phenocopies Rhd2-. Treatment of rhd2 roots with ROS partly suppresses the mutant phenotype and stimulates the activity of plasma membrane hyperpolarization-activated Ca2+ channels, the predominant root Ca2+ acquisition system. This indicates that NADPH oxidases control development by making ROS that regulate plant cell expansion through the activation of Ca2+ channels.

The primary root of Arabidopsis has a simple cellular organisation. The fixed radial cell pattern results from stereotypical cell divisions that occur in the meristem. Here we describe the characterisation of schizoriza (scz), a mutant with defective radial patterning. In scz mutants, the subepidermal layer (ground tissue) develops root hairs. Root hairs normally only form on epidermal cells of wild-type plants. Moreover, extra periclinal divisions (new wall parallel to surface of the root) occur in the scz root resulting in the formation of supernumerary layers in the ground tissue. Both scarecrow (scr) and short root (shr) suppress the extra periclinal divisions characteristic of scz mutant roots. This results in the formation of a single layered ground tissue in the double mutants. Cells of this layer develop root hairs, indicating that mis-specification of the ground tissue in scz mutants is uncoupled to the cell division defect. This suggests that during the development of the ground tissue SCZ has two distinct roles: (1) it acts as a suppressor of epidermal fate in the ground tissue, and (2) it is required to repress periclinal divisions in the meristem. It may act in the same pathway as SCR and SHR.

Transgenic tobacco genotypes expressing the maize Cat2 gene were developed with altered catalase (CAT) levels that resulted in a moderate increase of CAT activity in two transgenic lines. Bacterial infection, with a pathogen that does not share homology with the transgene, caused local and systemic down-regulation of the steady state mRNA levels of the 35S-driven transgene in a manner resembling post-transcriptional gene silencing (PTGS). Phenotypic symptoms of hypersensitive response (HR) and systemic acquired resistance (SAR) were similar in control SR1 and the transgenic genotypes. Induction of hin1, used as a molecular marker of plant responses to invading bacteria, displayed a similar pattern between control and transgenic lines, but some variation in the levels of expression was observed. The major difference was recorded in the ability of the plants to restrict bacterial growth during HR. All transgenic lines were more sensitive than control SR1, with two lines exhibiting a significantly reduced capacity to inhibit bacterial growth. This is consistent with the putative enhanced capacity of transgenic lines containing the maize Cat2 gene to more effectively remove H2O2, which may act as a direct antimicrobial agent. Steady state mRNA levels of PR-1 and PR-5 varied among the genotypes, possibly indicating differences in strength of the SAR signal. Transgenic line 2, which was the most sensitive during HR, was most effective in restricting bacterial growth during SAR. This indicates that a reverse correlation might exist between the severity of infection during HR and the ability to inhibit bacterial growth during SAR. Growth under high light conditions affected plant-pathogen interactions in control SR1, as well as in transgenic line 8. Early induction and higher expression of PR-1 and PR-5 was detected in both SR1 and line 8 in high light-grown plants as compared with their low light-grown counterparts. Our data indicate that growth under high light conditions can predispose plants to better resist pathogen attack, and may amplify local and systemic defense signals. Finally, one transgenic line, which exhibited 1.3-fold higher average CAT activity in comparison with the untransformed SR1 control, suffered significantly less methyl viologen (MV) damage than untransformed control plants at moderate and high MV concentrations.

Ion transport is important for driving nutrient transport across the syncytiotrophoblast and yet is poorly understood. We have examined K(+)currents under basal conditions in cultured cytotrophoblast cells, at various stages of differentiation, using the whole cell patch clamp technique. Cytotrophoblast cells were isolated from human term placenta and maintained in culture for up to 3 days. Cells were studied at four stages of progressive morphological differentiation: (i) mononuclear cells, (ii) mononuclear cells in aggregates, (iii) small multinucleate cells and (iv) large multinucleate syncytiotrophoblast-like cells. In the conditions of whole cell recording the only K(+) selective current identified in all cell types was a strong inwardly rectifying current which was sensitive to Ba(2+) and Cs(+). This current was unaffected by intracellular ATP whereas intracellular GTPgammas caused either run down of the current or activated a linear current. The characteristics of the current described are consistent with those of the inwardly rectifying K(+) channel Kir2.1. The inwardly rectifying K(+) current was observed in three out of 19 (16 per cent ) mononuclear cells, seven out of 21 (33 per cent ) mononuclear aggregates, eight out of 21 (38 per cent ) small multinucleate cells and 16 out of 19 (84 per cent ) large multinucleate cells. This inwardly rectifying K(+) current is likely to have an important role in determining net K(+) diffusion across the syncytiotrophoblast cell membrane, perhaps increasing in importance as the cells terminally differentiate., (Copyright 2001 Harcourt Publishers Ltd.)

The aims of this study were to establish the sites of mRNA expression of both the cystic fibrosis (CF) and multidrug resistance (MDR1) genes in human placental sections from early pregnancy (first, early and mid-second trimesters). Riboprobes specific for each of these two genes were generated and used for in situ hybridization experiments. The results show parallel mRNA expression for the CF and MDR1 genes, with the signal detected in the syncytiotrophoblast and cytotrophoblast cells of the placental villi. Other cell types within the villous core were negative. Similar results were obtained at all stages of pregnancy studied., (Copyright 1999 W.B. Saunders Company Ltd.)

Antioxidant enzymes function to eliminate reactive oxygen species (ROS) produced as a consequence of normal metabolic functions as well as environmental stress. In these studies, the responses of catalase (Cat), superoxide dismutase (Sod) and glutathione S-transferase (Gst), as well as D-ribulose-1,5-bisphosphate carboxylase/oxygenase (RbcS) genes were analyzed in 9- and 15-day postimbibition maize seedlings exposed to various ozone (O3) concentrations and time periods. After a single (acute) 6 h exposure, or 3, 6 and 10 consecutive days (chronic) exposure to O3, Cat1, Cat3, Gst1, Sod3, Sod4 and Sod4A transcript levels generally increased, while Cat2, RbcS and Sod1 levels decreased. Such changes in mRNA levels do not necessarily reflect parallel changes in the protein products of these genes. Changes in transcript levels seemed to be correlated with the spatial location of the isozymes encoded by the genes. The results are discussed with respect to gene regulation and expression, and the localization and function of these antioxidant enzymes during ozone-mediated oxidative stress.

The aim of this work was to determine the frequency of a base substitution (C-->T) identified in the Kir2.1 gene (approved gene symbol: KCNJ2; OMIM number: 600681). Polymerase chain reaction (PCR) of the area of the Kir2.1 gene containing this substitution was performed on 52 genomic DNA samples. Using single strand conformation polymorphism (SSCP) analysis, the genotype and allele frequencies were subsequently determined and the polymorphism identified in this study was verified by cycle sequencing. The data demonstrate that the C-->T nucleotide change identified corresponds to a silent polymorphism with a relatively high frequency. The deduced genotype frequencies of homozygotes and heterozygotes were: C/C: 73%; T/T: 2% and C/T: 25%. The deduced allele frequencies were C: 85.6% and T: 14.4%., (Copyright 1998 Academic Press.)

The aim of this study was to investigate whether the Kir2.1 gene is expressed by the human placenta throughout pregnancy and in cytotrophoblast cells at different stages of differentiation in culture. RNA was extracted from cytotrophoblast cells isolated from term placentas and maintained in culture for 18, 66 and 114 h and from first, second and third trimester placentas. Using the reverse transcriptase-polymerase chain reaction (RT-PCR) with gene-specific primers, a cDNA product of 1.2 kb, as expected for Kir2.1 gene expression, was detected in all the RNA samples from cytotrophoblast cells and from placentas. The RT-PCR products were verified by sequencing and by detection of the expected transcript size for the Kir2.1 mRNA at 5.6-5.7 kb on Northern blots, using the 1.2 kb cDNA generated by RT-PCR. Northern blot quantification, using a control 28S rRNA probe, showed no significant difference in Kir2.1 mRNA expression between any of the three stages of cytotrophoblast cell differentiation studied (ANOVA; n = 3 RNA samples from each stage). These data demonstrate that the Kir2.1 gene is expressed by the human placenta and, specifically, by cytotrophoblast cells, at all stages of development and differentiation.

The aims of this study were to establish whether both the cystic fibrosis (CF) and multidrug resistance (MDR1) genes are expressed in the human placenta during development and differentiation. To study their pattern of expression during development, RNA was extracted from first, second and third trimester human placentas. To investigate differentiation, RNA was extracted from cytotrophoblast cells isolated from human term placentas and maintained in culture for 18, 66, 90 and 114 h and from the undifferentiated choriocarcinoma cell line JAr. Using the reverse transcriptase-polymerase chain reaction (RT-PCR) with gene specific, intron spanning primers, a cDNA product of 1 kb, as expected for CF expression, was detectable following 35 cycles of PCR from all RNA samples except those from JAr; in the latter a product was only detected in one sample out of four separate passages and this was only just detectable after 40 cycles of PCR. RT-PCR using MDR1 specific primers resulted in a product from all samples at 0.34 kb as expected if this gene is expressed. These results demonstrate that both the CF and MDR1 genes are expressed in the human placenta at all stages of development and differentiation, although the expression of the CF, but not the MDR1, gene appears to be much weaker in the undifferentiated JAr cells in comparison with cytotrophoblast cells.

A transcriptional enhancer which has a consensus binding sequence for transcription enhancer factor-1 (TEF-1) has been found 3' of the hPL(3) gene. We examined whether TEF-1 is expressed by the human placenta and whether such expression is co-ordinated with that of human placental lactogen (hPL). Probing Northern blots of total RNA from first trimester and term placenta, the choriocarcinoma-derived cell line JAr and primary cultured cytotrophoblast cells with a cDNA for TEF-1 revealed transcripts of 12-13 kb and 3-4 kb. The level of TEF-1 expression was the same in first trimester as compared with term placenta and in undifferentiated JAr as compared with differentiated cytotrophoblast cells. hPL expression was tenfold higher in term compared with first trimester placenta and, whilst detectable in cytotrophoblast cells, was undetectable in JAr cells. These data show that TEF-1 is expressed by the placenta but is not co-ordinated with hPL expression.

In Azotobacter vinelandii, the anfHDGK operon encodes the subunits for the third nitrogenase complex. Two open reading frames (orf1 and orf2) located immediately downstream of anfK were shown to be required for diazotrophic growth under Mo- and V-deficient conditions. We have designated orf1 and orf2 anfO and anfR, respectively. Strains (CA115 and CA116) carrying in-frame deletions in anfO and anfR accumulate the subunits for nitrogenase 3 under Mo-deficient diazotrophic conditions. AnfO and AnfR are required for nitrogenase 3-dependent diazotrophic growth and 15N2 incorporation but not for acetylene reduction. AnfO contains a putative heme-binding domain that exhibits similarity to presumed heme-binding domains of P-450 cytochromes. Amino acid substitutions of Cys-158 show that this residue is required for fully functional AnfO as measured by diazotrophic growth under Mo- and V-deficient conditions. The nucleotide sequence of the region located immediately downstream of anfR has been determined. A putative rho-independent transcription termination site has been identified 250 bp from the 3' end of anfR. A third open reading frame (orf3), located downstream of anfR, does not appear to be required for diazotrophic growth under Mo- and V-deficient conditions.

The pattern of type VI collagen deposition in human endometrium throughout the menstrual cycle and in decidua of the first trimester of pregnancy was studied. Immunohistochemical analysis demonstrated a dense microfibrillar network of collagen VI in the stroma of preimplantation endometrium which was reduced during the peri-implantation period and no longer detected in first trimester decidua. However, type VI collagen was consistently present within blood vessel walls in both endometrium and decidua. Using in situ hybridization, mRNAs encoding alpha 1(VI), alpha 2(VI) and alpha 3(VI) chains within endometrial stromal and vascular cells were identified. All three mRNA species are abundant in the villous mesenchyme of the first trimester placenta. The detection of collagen VI mRNA species in the endometrium throughout the menstrual cycle suggests that the apparent decrease in abundance of extracellular immunoreactive fibrils may be a consequence of translational control, matrix redistribution or turnover. In contrast, in the first trimester of pregnancy, collagen VI protein was mainly absent from the decidual stroma and amounts of mRNA were very low, indicating a significant reduction in production. Loss of stromal type VI collagen contributes to the remodelling of the maternal extracellular matrix of pregnancy.

Two-dimensional gel electrophoresis of pea root and root hair proteins revealed the existence of at least 10 proteins present at elevated levels in root hairs. One of these, named RH2, was isolated and a partial amino acid sequence was determined from two tryptic peptides. Using this sequence information oligonucleotides were designed to isolate by PCR an RH2 cDNA clone. In situ hybridization studies with this cDNA clone showed that rh2 is not only expressed in root hairs, but also in root epidermal cells lacking these tubular outgrowths. During post-embryonic development the gene is switched on after the transition of protoderm into epidermis and since rh2 is already expressed in a globular pea embryo in the protoderm at the side attached to the suspensor, we conclude that the expression of rh2 is developmentally regulated. At the amino acid level RH2 is 95% homologous to the pea PR protein I49a. These gene encoding I49a is induced in pea pods upon inoculation with the pathogen Fusarium solani [12]. We postulate that rh2 contributes to a constitutive defence barrier in the root epidermis. A similar role has been proposed for chalcone synthase (CHS) and chitinase, pathogenesis-related protein that are also constitutively present in certain epidermal tissues.

Two cDNA clones of the soybean early nodulin GmENOD55 were characterized. These clones may represent two members of the soybean early nodulin gene family GmENOD55. GmENOD55 has an N-terminal signal peptide and it contains an internal domain consisting of proline and serine residues. Analyses of nodules lacking infection threads and intracellular bacteria suggest that the GmENOD55 gene is first expressed after release of Bradyrhizobium japonicum in plant cells. This conclusion is supported by in situ hybridization studies showing that the expression is restricted to the infected cell type.