Copper Uptake Efficiency and Its Distribution Within Bioenergy Grass Giant Reed.

To evaluate copper uptake and its toxicity on bioenergy grass giant reed (Arundo donax L.), experiments were carried out using two epigenetic clonal lines - American (BL) and Hungarian (20SZ) ecotypes - grown on elevated Cu concentrations up to 26.8 mg L(-1). Neither ecotype showed any noticeable foliar symptoms of Cu toxicity at concentrations tested up to 10 mg L(-1). Dry mass of plants of both ecotypes significantly increased at the highest Cu treatment compared to control. Although the BL ecotype had greater capacity to uptake Cu than 20SZ, the dry mass and shoot length of BL was higher than that of 20SZ. Values of bioconcentration and transportation factors were higher in the BL than in the 20SZ ecotype. Almost 45 % of total Cu content within the whole plant was found in the plant root of both ecotypes. This demonstrated both ecotypes can be utilized for Cu phytoremediation alongside with significant biomass production.

The frequency of Trp64Arg polymorphism of the beta3-adrenergic receptor gene in healthy and obese Hungarian children and its association with cardiovascular risk factors.

OBJECTIVE: To estimate the frequency of Arg64 allele of the beta(3)-adrenergic receptor (3-BAR) gene in healthy (H) and obese (O) Hungarian children, and to look for possible associations between this polymorphism and some clinical and metabolic characteristics of obese children. PATIENTS/METHODS: In all, 147 healthy (male: 68) and 295 obese (male: 168) children were examined. The average age of the children in the two groups was 12.4+/-1.7 vs 12.6+/-3.2, respectively. Exon 1 of 3-BAR was amplified by polymerase chain reaction and the fragments were digested with BstN1. In obese children, oral glucose tolerance test was carried out and blood pressure (BP) was checked. RESULTS: The frequency of Trp64Arg polymorphism in normal and obese Hungarian children was similar (H vs O: n=14/9.5% vs n=35/11.8%). Obese children carrying the Arg64 allele (n=35, male: 23) were compared to randomly chosen, obese children without the Arg64 allele (n=35, male: 20). A significant difference was found between the body weight (81.2+/-23.2 vs 75.6+/-17.7 kg; mean+/-s.d.; P<0.01), body fat (38.8+/-3.9 vs 36.5+/-2.3%; mean+/-s.d.; P<0.05), mean fasting insulin levels (31.4+/-16.7 vs 16.9+/-7.6 microIU/ml; P<0.001) and mean systolic BP values (125.2+/-10.1 vs 114.5+/-8.3 mmHg; P<0.001) of the two obese groups. CONCLUSIONS: The frequency of Trp64Arg polymorphism was similar in Hungary as compared to other European countries. Although the prevalence of this polymorphism was similar in H and O children, the presence of Arg64 allele seems to be associated with increased adiposity, elevated systolic BP and higher fasting insulin levels.

Differential mercury volatilization by tobacco organs expressing a modified bacterial merA gene.

Mercury pollution is a major environmental problem accompanying industrial activities. Most of the mercury released ends up and retained in the soil as complexes of the toxic ionic mercury (Hg2+), which then can be converted by microbes into the even more toxic methylmercury which tends to bioaccumulate. Mercury detoxification of the soil can also occur by microbes converting the ionic mercury into the least toxic metallic mercury (Hg0) form, which then evaporates. The remediation potential of transgenic plants carrying the MerA gene from E. coli encoding mercuric ion reductase could be evaluated. A modified version of the gene, optimized for plant codon preferences (merApe9, Rugh et al. 1996), was introduced into tobacco by Agrobacterium-mediated leaf disk transformation. Transgenic seeds were resistant to HgCl2 at 50 microM, and some of them (10-20% ) could germinate on media containing as much as 350 microM HgCl2, while the control plants were fully inhibited or died on 50 microM HgCl2. The rate of elemental mercury evolution from Hg2+ (added as HgCl2) was 5-8 times higher for transgenic plants than the control. Mercury volatilization by isolated organs standardized for fresh weight was higher (up to 5 times) in the roots than in shoots or the leaves. The data suggest that it is the root system of the transgenic plants that volatilizes most of the reduced mercury (Hg0). It also suggests that much of the mercury need not enter the vascular system to be transported to the leaves for volatilization. Transgenic plants with the merApe9 gene may be used to mercury detoxification for environmental improvement in mercury-contaminated regions more efficiently than it had been predicted based on data on volatilization of whole plants via the upper parts only (Rugh et al. 1996).

Isodicentric Y chromosome in an Ullrich-Turner patient without virilization.

We report on a 17-year-old young woman with Ullrich-Turner syndrome (UTS), who was found to have a karyotype 45,X/46,X,idic(Y)(q11). She had age-appropriate genitalia without virilization in spite of the presence of the Y-derived marker chromosome and SRY locus in 70% of her lymphocytes. Having reviewed the literature, we conclude that a possible explanation for the lack of virilization in these mosaic patients is most likely an uneven distribution of tissue mosaicism (gonadal mosaicism).

Distal deletion, del(2)(q33.3q33.3), in a patient with severe growth deficiency and minor anomalies.

We report on an 18-month-old boy with a 2q33.3 deletion. The clinical findings observed in the propositus included minor anomalies of face and distal limbs, intrauterine and postnatal growth retardation, microcephaly and, so far, moderate developmental delay. Conventional GTG banded chromosome analysis indicated a small deletion in distal 2q. Subsequent analysis by fluorescent in situ hybridization (FISH) using different probes allowed us to narrow down the deletion to most or all of segment 2q33.3. This case shows the importance of the application of different YAC probes for a precise determination of breakpoints in small interstitial deletions.

A heartwood pigment in Dalbergia cell cultures.

In an extensive survey of the genera Baphia, Caesalpinia, Dalbergia, Haematoxylon, and Pterocarpus, we have identified a number of species whose cell cultures accumulated pigments similar to those in heartwood. Thirteen rosewood (Dalbergia) species produced a purple quinonemethide pigment in the callus that was apparently identical between the species. The pigment was first purified from D. retusa cell culture and its structure was elucidated by mass, infrared, and detailed 1H and 13C NMR and NOE spectroscopic studies including 2D experiments (COSY, NOESY, HMQC, and HMBC). Retusapurpurin A (1a) is a C(30) isoflavan of novel skeleton whose formation can be rationalized to occur via regioselective oxidative coupling of an isoflavan to 4,4'-dihydroxy-2'-methoxychalcone. Retusapurpurin A was also isolated from D. parviflora heartwood and cell culture indicating that stress metabolism pathways that are shared with heartwood-type secondary metabolism subpathways are initiated in Dalbergia cell cultures. Therefore, Dalbergia cell cultures afford a good model system for studying heartwood-type metabolic differentiation.

Participation of "emotionality" in activation of the adrenal cortex of rats in some stress situations.

The present work attempts to interpret the relation between humoral and behavioral manifestations in rats under conditions of stress. The index of the humoral reaction to stress was plasma corticosterone levels. Initially, behavioral manifestations which are commonly subsumed under the term "emotionality" were studied. The methodological problems connected with measurement of emotionality led us to adopt instead the concept of "non-specific excitability" as proposed by J. Lat. Results are presented which point to a relationship between excitability of the animals and the activity of their adrenal cortex.

Tissue specific mosaicism of trisomy 9 in a patient with severe torsion scoliosis.

We report on a female patient who had mosaic trisomy 9, presenting with severe scoliosis and mental retardation. Scoliosis is seldom reported in patients with mosaic trisomy 9 syndrome. FISH studies in our proband detected no trisomic cell line in the paravertebral muscle.

A girl with cutaneous hyperpigmentation, café au lait spots and ring chromosome 15 without significant deletion.

Ring chromosome 15 [r(15)] syndrome is characterised by specific facial features, café au lait spots, failure to thrive, mental retardation and typically with a terminal deletion of the long arm of chromosome 15. We report a 2.5 year old girl showing normal growth and development, large hyperpigmented skin changes showing hypopigmentated areas inside, multiple café au lait spots and premature graying-like hypopigmentation of scalp hair. She had a karyotype of r(15) in peripheral lymphocytes and fibroblasts. By FISH analysis the breakpoint was located distal to locus D15S936 (15q26.3) and within 300 kb of the end of the chromosome, indicating no deletion of functional genes on 15q. Hyperpigmentation and café au lait spots are rare signs in ring chromosome syndromes, but with r(15) syndrome, café au lait spots have been described in about 30% of patients and have been considered to result from the deletion of gene(s) on distal 15q. Based on the frequent observation of patchy hyperpigmentation with the r(15) syndrome, absent hyperpigmentation in cases of distal 15q deletion without a ring chromosome, and the telomeric breakpoint location in our patient indicating no significant deletion, we propose that the cutaneous hyperpigmentation and café au lait spots in our proband represent effects of the r(15) chromosome but are not caused by the deletion of specific gene(s) on distal 15q. Patchy skin hypopigmentation is a well known nonspecific sign in cytogenetic mosaicism which is commonly seen in ring syndrome.

Genetic diversity in Proteus penneri.

DNA of thirteen Proteus penneri strains derived from four European countries (nine strains from Germany, two strains from United Kingdom, one strain from Turkey, one strain from Hungary) was examined by random amplified polymorphic DNA-PCR (RAPD-PCR) method. RAPD with primer AACGCGCAAC gave different patterns, which suggests a DNA sequence microdiversity within this species. The method provides a fast, economical and reproducible means for typing P. penneri.

[22q11.2 chromosome deletion and velo-cardio-facial syndrome in a patient with tetralogy of Fallot]. / 22q11.2 kromoszómadeléció és velo-cardio-facialis szindróma egy Fallot-tetralógiás betegben.

Velo-cardio-facial syndrome includes the following clinical features: congenital heart anomaly, velo-pharyngeal malformations and dysmorphic facial features. In 80% of the patients microdeletion of the long arm of chromosome 22 can be detected. Many patients with DiGeorge syndrome have the same underlying chromosomal etiology, however the same deletion results in severe dysmorphic features, immune defect, hypocalcaemia and in severe mental retardation. In isolated heart defects deletion can be detected in about 15% of the patients. We report the detection of the microdeletion in a patient with tetralogy of Fallot, minimal dysmorphic facial features and normal psychomotor development. Early diagnosis of 22q11.2 chromosomal deletion is important for genetic counseling and further rehabilitation. We emphasize the importance of genetic screening in patients with isolated congenital heart defects.

[Molecular biologic screening test (PCR) for fragile X syndrome]. / Fragilis X szindróma szürésére alkalmas molekuláris biológiai (PCR) eljárás.

Fragile X syndrome is the most common inherited from of familial mental retardation. It is caused by an expanded CGG repeat in the first exon of the fragile X mental retardation gene. A polymerase chain reaction based technique was used for the identification of full mutations among men. According to our conditions full mutations failed to amplify. An internal control was used at a CG rich region 147 bp upstream of the polymorphic region. The bands were visualised on silver stained polyacrylamide gels. From the 57 individuals studied molecular analysis was performed on 38 males and 16 females. From the 26 males with suspected fragile X syndrome 9 males resulted in no amplification of the 500 kb product, all having a positive cytogenetic result for fragile X syndrome. One cytogeneticly positive male had normal results by molecular studies suggesting a different mutation. All control males had normal results. The results on the 16 females studied were inconclusive. We suggest that our method is highly sensitive and specific for screening males for fragile X syndrome.

[Vascular diseases, spina bifida and schizophrenia in a single family associated with the heterozygote mutation of the heat-sensitive variant of methylenetetrahydrofolate reductase]. / Errendszeri betegségek, spina bifida és schizophrenia egy családon belüli együttes elófordulása a metilén-tetrahidrofolsav-reduktáz enzim hóérzékeny variánsának heterozigóta mutációjával.

Homozygous mutation of the thermolabile variant of methylene tetrahydrofolate reductase (MTHFR) may result in hyperhomocystinemia, leading to an increased risk for early cardiovascular disease, neural tube defects, and possibly major depression, schizophrenia. According to recent studies heterozygosity for the thermolabile variant of the MTHFR gene mutation is also more frequent in patients with thrombotic disease compared to that in the average population. We report on a family with different types of early vascular disease. In four consecutive generations MTHFR heterozygosity was detected: in the proband and in her mother, grandfather and daughter. Further conditions of the family members, possibly due to carrying the mutation, came to light by the pedigree analysis and examinations. The patient had pulmonary emboli at young age, her aunt died of spina bifida shortly after birth. The patient's mother suffers from schizophrenia and depression. The grandfather had pulmonary emboli, her sister with spina bifida occulta also carries the same mutation, as does her daughter who is sofar asymptomatic. In other asymptomatic members of the family no mutations were found. Unexpectedly, hyperhomocystinemia was detected in all heterozygote individuals. Our study demonstrates the necessity for folic acid therapy in mutation carriers to prevent early vascular events, depression and schizophrenia, and also to reduce the risk for neural tube defects in a preconception setting.

[Kennedy disease in a patient with progressive speech disorder]. / Kennedy-betegség egy progresszív beszédzavarban szenvedó férfiben.

Kennedy disease is an adult onset neuromuscular disease characterized by slowly progressive proximal and bulbar muscle weakness. The disease associates with gynecomastia, adult onset infertility and sensory neuropathy, and caused by pathologic expansion of CAG repeats at the N-terminal region of the androgen-receptor gene at Xq11-q12. We report on a patient presenting with slowly progressive muscle weakness of the lower extremities, progressive dysartry and swallowing difficulties. The clinical symptoms were not fully specific for the disease. Moreover the family history was suggestive for an autosomal dominant trait meaning a diagnostic pitfall at the original examination. Finally the firm diagnosis of the Kennedy disease was established by a polimerase chain reaction based method.

Phytoaccumulation potentials of two biotechnologically propagated ecotypes of Arundo donax in copper-contaminated synthetic wastewater.

An in vitro experiment was carried out to evaluate the phytoremediation potentials of two somatic embryo-derived ecotypes of Arundo donax-BL (American ecotype) and 20SZ (Hungarian ecotype)-of copper from synthetic wastewater. The two ecotypes were grown under sterile conditions in tubes containing a nutrient solution supplied with increasing doses of Cu (0, 1, 2, 3, 5, 10, and 26.8 mg L(-1)) for 6 weeks. The translocation and bioaccumulation factors and removal rate were estimated. In general, increasing Cu concentration in nutrient solution slightly decreased root, stem and leaf biomass without toxicity symptoms up to 26.8 mg L(-1). Moreover, both ecotypes showed high Cu removal efficiency from aqueous solution. However, Cu removal rate ranged between 96.6 to 98.8% for BL ecotype and 97 to 100% for 20SZ ecotype. Data illustrated that both BL and 20SZ ecotypes may be employed to treat Cu-contaminated water bodies up to 26.8 mg L(-1).

Expression of DNA coding for diphtheria toxin chain a is toxic to plant cells.

DNA coding for the enzymatically active subunit A of diphtheria toxin was placed under the control of the cauliflower mosaic virus 35S promoter and the Agrobacterium left transfer-DNA gene 7 polyadenylation signal. Agrobacteria carrying a binary plant vector with the chimeric diphtheria toxin A gene had very low transforming activity in tobacco (Nicotiana tabacum L.), and greatly diminished the recovery of stable transformants when mixed together with agrobacteria which alone transformed plant cells well. The introduction of this chimeric molecule into tobacco cells by electroporation lowered the level of the transient expression of the coelectroporated chloramphenicol acetyltransferase reporter gene indicating that expression of diphtheria toxin chain A in plant cells is toxic. We have developed a binary vector pGA987 which can be used for probing a variety of plant promoters.

Independent integration and seed-transmission of the TR-DNA of the octopine Ti plasmid pTi Ach5 in Nicotiana plumbaginifolia.

After co-cultivation of diploid Nicotiana plumbaginifolia protoplasts with an octopine-type Agrobacterium tumefaciens strain (LBA 4013) putative transformants were selected for hormone-independent growth, and were tested for T-DNA markers. The number of transformants expressing only TL-DNA markers, i.e. phytohormone autotrophy and octopine synthase, was an order of magnitude higher than that of the cell lines which were simultaneously positive for both TL- and TR-DNA markers (the latter being mannopine and agropine). In one transformant, line no. 101, only the TR-DNA markers were found. Not each of the TL-, or TR-DNA markers were expressed in each transformant resulting in a variety of phenotypes. It included the unorganized or the shoot-teratoma type of growth combined with the presence or absence of opines; e.g. agropine was absent from some of the transformants containing its precursor, mannopine. 5-Azacytidine did not induce agropine synthesis in these lines. Southern blot analysis showed that the TR-DNA region coding for agropine synthesis was rearranged or absent in one of these lines. Similar variation in the expression of agropine and mannopine production was observed in transformants obtained with the leucinopine-type strain A281.From line 101 plants could be easily regenerated with the ability to synthesize agropine and mannopine. The segregation in the self-progeny fitted to a 3:1 ratio, indicating that the TR-DNA was carried by a single chromosome. The Southern blot analysis showed that only opine-positive plants contained TR-DNA. It also confirmed the absence of the TL-DNA, demonstrating the independent integration of the TR-region of the octopine-type Ti plasmid pTi Ach5.