Biodegradation of 4-nitroanisole by two Rhodococcus spp.

Two Rhodococcus strains, R. opacus strain AS2 and R. erythropolis strain AS3, that were able to use 4-nitroanisole as the sole source of carbon and energy, were isolated from environmental samples. The first step of the degradation involved the O-demethylation of 4-nitroanisole to 4-nitrophenol which accumulated transiently in the medium during growth. Oxygen uptake experiments indicated the transformation of 4-nitrophenol to 4-nitrocatechol and 1,2,4-trihydroxybenzene prior to ring cleavage and then subsequent mineralization. The nitro group was removed as nitrite, which accumulated in the medium in stoichiometric amounts. In R. opacus strain AS2 small amounts of hydroquinone were produced by a side reaction, but were not further degraded.     

Vascular Development and Sap Flow in Apple Pedicels

Xylem and phloem tissues of the pedicel of apple fruit increasein cross-sectional area throughout development. The increasein phloem is similar in the two cultivars examined (Cox's OrangePippin and Royal Gala) and reflects a steadily increasing phloemsap flow to the fruit. The increase in xylem tissue is due toa proliferation of non-conducting, structural, components sinceclose examination reveals no increase in the number of vesselelements from just after flowering onwards. The greater number,and the larger diameter, of the vessels in Cox's explains theinitially higher xylem conductance found in this cultivar. In vitro measurements of xylem exudation reveal a decline duringthe growing season in the xylem conductance of both cultivarsand an increasing proportion of fruit (particularly in Cox's)in which the xylem comes to be totally non-conducting. Thisobservation is in line with previously reported measurementsof xylem sap flow in vivo. The straightforward techniques used in this study offer a feasiblealternative to more arduous methods of assessing xylem and phloemsap flows and their balance during growth.Copyright 1994, 1999Academic Press Apple, xylem, phloem, vascular development, sap flow, Malus domestica Borkh     

Machado-Joseph Disease in Pedigrees of Azorean descent is Linked to Chromosome 14

A locus for Machado-Joseph disease (MJD) has recently been mapped to a 30-cM region of chromosome 14q in five pedigrees of Japanese descent. MJD is a clinically pleomorphic neurodegenerative disease that was originally described in subjects of Azorean descent. In light of the nonallelic heterogeneity in other inherited spinocere-bellar ataxias, we were interested to determine if the MJD phenotype in Japanese and Azorean pedigrees arose from mutations at the same locus. We provide evidence that MJD in five pedigrees of Azorean descent is also linked to chromosome 14q in an 18-cM region between the markers D14S67 and AACT (multipoint lod score +7.00 near D14S81). We also report molecular evidence for homozy-gosity at the MJD locus in an MJD-affected subject with severe, early-onset symptoms. These observations confirm the initial report of linkage of MJD to chromosome 14; suggest that MJD in Japanese and Azorean subjects may represent allelic or identical mutations at the same locus; and provide one possible explanation (MJD gene dosage) for the observed phenotypic heterogeneity in this disease.     

Multiplex PCR for detection of the heat-labile toxin gene and shiga-like toxin I and II genes in Escherichia coli isolated from natural waters.

A triplex PCR method was developed to simultaneously amplify a heat-labile toxin sequence (LT) of 258 bp, a shiga-like toxin I sequence (SLT I) of 130 bp, and a shiga-like toxin II sequence (SLT II) of 346 bp from toxigenic strains of Escherichia coli. This method was used to screen 377 environmental E. coli isolates from marine waters or estuaries located in Southern California and North Carolina for enterotoxigenic or enterohemorrhagic E. coli strains. Of the 377 E. coli screened, one isolate was found to belong to the enterotoxigenic group, since it contained a LT homologous sequence, and one isolate was found to belong to the enterohemorrhagic group, since it contained a SLT I homologous sequence. None was found to contain SLT II homologous sequences. The pathogenicity of the positive environmental E. coli isolates was confirmed by standard bioassays with Y-1 adrenal cells and Vero cells to confirm toxin production. Our results suggest that toxigenic E. coli occurs infrequently in environmental waters and that there is a low public health risk from toxigenic E. coli in coastal waters.     

Terminal electron transport system (ETS)-activity in the sediment of Lake Balaton,Hungary

Terminal electron transport system (ETS)-activity of the sediment and plankton of Lake Balaton, the largest shallow lake of Central Europe was measured by tetrazolium-reduction biweekly during 1989–1990 and in the spring of 1991. Sediment proved to be enzymatically active to 30-35 cm down in the hypertrophic Keszthely Bay and to 15–20 cm down in the meso-eutrophic Siófok Basin. Sediment ETS-activity exceeded planktonic activity 15 to 24 fold.The total activity m^–2 showed one or two order of magnitude higher respiratory potential in Lake Balaton than needed for complete oxidation of the planktonic primary production; most of this potential was detected in the upper 3–5 cm sediment layer in springs. Incubations of cell-free homogenates of sediment bacteria showed that ETS remains active days after death of organisms at low temperature. Accumulated postmortem ETS-activity derived from the benthic diatoms, bacteria, plankton deposit and dead summer macrophytes seems to be responsible for the high ETS-activity of the sediment in the warming periods in springs. These enzyme fractions may contribute to the rapid oxidation of the alkaline, well-aerated lake.     

Degradation by and toxicity to bacteria of chlorinated phenols and benzenes,and hexachlorocyclohexane isomers

Mixed cultures degrading chlorinated benzenes, chlorinated phenols, or hexachlorocyclohexane (HCH) as the sole source of carbon and energy were obtained by enrichment from contaminated soil samples. Cultures which metabolized 3-chlorophenol (3-CP), 2,3-dichlorophenol (2,3-DCP), or 2,6-dichlorophenol (2,6-DCP) were able to utilize several other chlorinated compounds as substrates, whereas cultures enriched with 1,2,4,5-tetrachlorobenzene (1,2,4,5-TeCB), -HCH, or -HCH did not metabolize most of the other chlorinated congeners tested. Chloride release and growth rates with all four chlorinated phenols decreased with increasing initial substrate concentrations within the range of 30–250 mol liter^–1. Maximum chloride release was 3.8 mg liter^–1 corresponding to 35 mol liter^–1 trichlorophenol within 7 weeks. In contrast, the rate of metabolism of the nonphenolic compounds 1,2,4,5-TeCB, -HCH, or -HCH increased with increasing substrate concentrations. Initial concentrations of 750 mol liter^–1 -HCH or 1,2,4,5-TeCB were completely dechlorinated within 2 weeks. Because aqueous solubility and bioavailability of the chlorophenolic compounds is much higher than that of the nonphenolic compounds, it is suggested that the high bioavailability of the chlorophenolic compounds is the reason for the high toxicity of these substrates to the degrading cultures. In contrast, the low aqueous solubilities of the chlorinated benzenes and HCH-isomers caused consistently low concentrations in the medium, which were high enough to induce degradation but too low to damage the bacterial cells.Correspondence to: E. Lang     

Interfacial and micellar behavior of glucose lipid

Summary Glucose lipid of notable surfactant properties was produced by using hydrocarbon assimilating bacterium of the bacterial strain MM1. Its surface active properties were notable in spite of ,-hydrophilic and bulky molecule. The critical micelle concentration (cmc) was small (0.165 M); surface and interfacial tension for hexadecane at 0.1 % (pH 7.35) solution, 24.6 and 13.3 mN/m, respectively. The emulsifying action was excellent and comparable to those of rhamnolipids. The micelle has the outer radius. 28.5 Å and inner core, 6.7 Å on the basis of the concentric spherical shell model by means of the small angle X-ray solution scattering measurement.Dedicated to Professor Fritz Wagner on the occasion of his 65th birthday.     

Morphological alterations and cytoskeletal reorganization in opossum kidney (OK) cells during osmotic swelling and volume regulation

Cells from a variety of tissues regulate their volume when exposed to anisotonic conditions. After exposure of cells to hypotonic conditions, the rapid phase of cell swelling is followed by a slower phase of cell shrinkage towards the initial volume. The present study investigates morphological alterations of adherent and fully spread cells after exposure to hypotonic conditions and the reorganization of cytoskeletal components such as F-actin, actin-binding proteins, microtubules and intermediate-sized filaments. We used cells of a continuous epithelial cell line from the opossum kidney (OK cells), which were exposed to hypotonic conditions for a period of 60 min at 25° C. The osmolarity was reduced by 40% from 320 mosmol/l (isotonic conditions) to 192 mosmol/l (hypotonic conditions). The initial swelling after exposure of OK cells to hypotonic conditions caused enhanced ruffling membrane activity, formation of lamellipodia and an extended space between adjacent cells which was caused by a more rounded cell shape. Moreover, the height of cells located in the centre of cell clusters increased by 32±8% (mean value±SEM) as checked by morphometric analysis of the vertical distance between the apical and basolateral F-actin domain. Although the fluorescence intensity and organization of F-actin in a horizontal direction remained unaltered during cell swelling, we observed a loss of periodicity and irregular distribution of myosin aggregates and a partial rear-rangement of vimentin filaments in the form of short fragments. In all experiments the organization of microtubules was observed to be unaltered. The alterations described above were reversible during cell shrinkage towards the initial volume, i.e. at 60 min after exposure to hypotonic conditions cell morphology and cytoskeletal organization no longer differed from the corresponding controls which were kept under isotonic conditions for the whole experimental period. The results demonstrate that only certain intracellular cytoskeletal components are actively involved in cell swelling and regulatory volume decrease.     

Midwife managed delivery unit: a randomised controlled comparison with consultant led care.

OBJECTIVE--To examine whether intrapartum care and delivery of low risk women in a midwife managed delivery unit differs from that in a consultant led labour ward. DESIGN--Pragmatic randomised controlled trial. Subjects were randomised in a 2:1 ratio between the midwives unit and the labour ward. SETTING--Aberdeen Maternity Hospital, Grampian. SUBJECTS--2844 low risk women, as defined by existing booking criteria for general practitioner units in Grampian. 1900 women were randomised to the midwives unit and 944 to the labour ward. MAIN OUTCOME MEASURES--Maternal and perinatal morbidity. RESULTS--Of the women randomised to the midwives unit, 647 (34%) were transferred to the labour ward antepartum, 303 (16%) were transferred intrapartum, and 80 (4%) were lost to follow up. 870 women (46%) were delivered in the midwives unit. Primigravid women (255/596, 43%) were significantly more likely to be transferred intrapartum than multi-gravid women (48/577, 8%). Significant differences between the midwives unit and labour ward were found in monitoring, fetal distress, analgesia, mobility, and use of episiotomy. There were no significant differences in mode of delivery or fetal outcome. CONCLUSIONS--Midwife managed intrapartum care for low risk women results in more mobility and less intervention with no increase in neonatal morbidity. However, the high rate of transfer shows that antenatal criteria are unable to determine who will remain at low risk throughout pregnancy and labour.     

Radiation-induced concomitant overexpression of p53, p62c-fos and p21N-fas in mouse epidermis

Abstract. Early molecular events in radiation carcinogenesis in vivo are difficult to study, especially because it is usually impossible to know in advance the exact location of a radiation-induced tumour. In the present study, we have attempted to overcome this difficulty by exposing a very small area of hairless mouse skin to high-dose beta radiation (i.e. immobilized hot particles) on the reasonable assumption that a malignant tumour would subsequently develop and be found at the exposed site in a long-term follow-up. The results showed that at an exposed site, before the appearance of a visible or histologically detectable tumour, overexpression of the product of the tumour suppressor gene p53 was common (28% of the sites studied; tested with PAbl801, PAb421, PAb240 and a polyclonal antibody CM1) and that this change was regularly accompanied by overexpression of p62^c-fos and p21^N-ras. Expression of several other oncoproteins studied (p39^c-jun, p21^K-ras, p21^H-ras) was not altered at these sites. A similar pattern of changes was observed in a visible and histopathologically distinct tumour that was analyzed after its development at an exposed site. These results suggest a re'markably regular pattern of molecular changes, induced by ionizing radiation in mouse epidermis, which might be associated with carcinogenesis.