bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of green fluorescent protein and luciferase biomarkers to monitor survival and activity of Arthrobacter chlorophenolicus A6 cells during degradation of 4‐chlorophenol in soil
Authors: Elväng Annelie M. , Westerberg Karolina , Jernberg Cecilia , Jansson Janet K. 
Published: March 2003

The recently isolated novel species Arthrobacter chlorophenolicus A6 is capable of growth on and degradation of high concentrations of 4‐chlorophenol (up to 350 μg ml−1) as the sole carbon and energy source. This strain shows promise for bioremediation of environmental sites contaminated with high levels of chlorophenols. In this study, green fluorescent protein (gfp) or luciferase (luc) genes were used as biomarkers for monitoring cell number and activity, respectively, during degradation of 4‐chlorophenol by A. chlorophenolicus cells. The individual marked strains, Arthrobacter chlorophenolicus A6L (luc‐tagged) and Arthrobacter chlorophenolicus A6G (gfp‐tagged), were monitored during degradation of 250 μg ml−1 4‐chlorophenol in pure culture and 175 μg g−1 4‐chlorophenol in soil microcosms. Both gene‐tagged strains were capable of cleaning up the contaminated soil during 9 d incubation. During the bioremediation experiments, the luc‐tagged cells were monitored using luminometry and the gfp‐tagged cells using flow cytometry, in addition to selective plate counting for both strains. The cells remained at high population levels in the soil (evidenced by GFP‐fluorescent cell counts) and the A. chlorophenolicus A6L population was metabolically active (evidenced by luciferase activity measurements). These results demonstrate that the Arthrobacter chlorophenolicus A6 inoculum is effective for cleaning‐up soil containing high concentrations of 4‐chlorophenol.
In situ identification of polyphosphate‐ and polyhydroxyalkanoate‐accumulating traits for microbial populations in a biological phosphorus removal process
Authors: Liu Wen‐Tso , Nielsen Alex T. , Wu Jer‐Horng , Tsai Chin‐Sun , Matsuo Yoshitaka , Molin Søren 
Published: March 2003

Polyphosphate‐ and polyhydroxyalkanoate (PHA)‐accumulating traits of predominant microorganisms in an efficient enhanced biological phosphorus removal (EBPR) process were investigated systematically using a suite of non‐culture‐dependent methods. Results of 16S rDNA clone library and fluorescence in situ hybridization (FISH) with rRNA‐targeted, group‐specific oligonucleotide probes indicated that the microbial community consisted mostly of the α‐ (9.5% of total cells), β‐ (41.3%) and γ‐ (6.8%) subclasses of the class Proteobacteria, Flexibacter–Cytophaga (4.5%) and the Gram‐positive high G+C (HGC) group (17.9%). With individual phylogenetic groups or subgroups, members of Candidatus Accumulibacter phosphatis in the β‐2 subclass, a novel HGC group closely related to Tetrasphaera spp., and a novel γ‐proteobacterial group were the predominant populations. Furthermore, electron microscopy with energy‐dispersive X‐ray analysis was used to validate the staining specificity of 4,6‐diamino‐2‐phenylindole (DAPI) for intracellular polyphosphate and revealed the composition of polyphosphate granules accumulated in predominant bacteria as mostly P, Ca and Na. As a result, DAPI and PHA staining procedures could be combined with FISH to identify directly the polyphosphate‐ and PHA‐accumulating traits of different phylogenetic groups. Members of Accumulibacter phosphatis and the novel gamma‐proteobacterial group were observed to accumulate both polyphosphate and PHA. In addition, one novel rod‐shaped group, closely related to coccus‐shaped Tetrasphaera, and one filamentous group resembling Candidatus Nostocoidia limicola in the HGC group were found to accumulate polyphosphate but not PHA. No cellular inclusions were detected in most members of the α‐Proteobacteria and the Cytophaga–Flavobacterium group. The diversified functional traits observed suggested that different substrate metabolisms were used by predominant phylogenetic groups in EBPR processes.
Biodiversity of Geodermatophilaceae isolated from altered stones and monuments in the Mediterranean basin
Authors: Urzì Clara , Brusetti Lorenzo , Salamone Paola , Sorlini Claudia , Stackebrandt Erko , Daffonchio Daniele 
Published: March 2003

An investigation was made into the occurrence and biodiversity of Geodermatophilaceae on 78 samples of altered stone surfaces from 24 monuments and natural stones in the Mediterranean basin; it was found that the total microbial counts ranged between 0 and 107 cfu g−1 dry weight. Members of the Geodermatophilaceae family were isolated from 22 of the 78 samples examined, with the incidence of Geodermatophilaceae colonies in the cultivable population ranging from 1% to 100%. The highest percentage was found in six samples of markedly deteriorated stone. Sixty‐five strains randomly isolated from the plates were clustered in six different groups by amplified 16S rDNA restriction analysis (ARDRA) using five different restriction enzymes. Twenty‐five strains, representing all the ARDRA haplotypes, were characterized further by partial sequencing (350–550 bp) of the 16S rDNA and by analysing 76 morphological, metabolic and physiological properties. The strains were associated with three well‐separated clusters of the genera Geodermatophilus, Blastococcus and Modestobacter. On the basis of 16S rDNA sequence and ARDRA analysis, only two strains were found to be related to the two reference strains of Geodermatophilus. All the others could be grouped with Blastococcus aggregatus (19 strains) or the Antarctic species Modestobacter multiseptatus (44 strains), suggesting that it is these two groups, rather than Geodermatophilus, that tend to colonize the stone surfaces, and that Modestobacter‐like strains are also found in temperate/Mediterranean climates. From the BOX‐polymerase chain reaction (PCR) data, it can be seen that the Modestobacter‐like strains, belonging to the most represented ARDRA haplotype (haplotype B, 34 strains), are very polymorphic and that, over a stone surface, there is a wide genetic diversity at the microsite level.
Microbial reduction of Fe(III) in the presence of oxygen under low pH conditions
Authors: Küsel Kirsten , Roth Ursula , Drake Harold L. 
Published: March 2003

SummaryIn acidic, coal mining lake sediments, facultatively anaerobic Acidiphilium species are probably involved in the reduction of Fe(III). Previous results indicate that these bacteria can co‐respire O2 and Fe(III). In this study, we investigated the capacity of the sediment microbiota to reduce Fe(III) in the presence of O2 at pH 3. In sediment microcosms with 4% O2 in the headspace, the concentration of Fe(II) increased at a rate of 1.03 μmol (g wet sediment)−1 day−1 within the first 7 days of incubation which was similar to the rate obtained with controls incubated under anoxic conditions. However, in microcosms incubated under air, Fe(II) was consumed after a lag phase of 8 h with a rate of 2.66 μmol (g wet sediment)−1 day−1. Acidiphilium cryptum JF‐5, isolated from this sediment, reduced soluble Fe(III) with either 4 or 21% O2 in the headspace, and concomitantly consumed O2. However, the rate of Fe(II) formation normalized for cell density decreased under oxic conditions. Schwertmannite, the predominant Fe(III)‐mineral of this sediment, was also reduced by A. cryptum JF‐5 under oxic conditions. The rate of Fe(II) formation by A. cryptum JF‐5 decreased after transfer from preincubation under air in medium lacking Fe(III). Acidiphilium cryptum JF‐5 did not form Fe(II) when preincubated under air and transferred to anoxic medium containing Fe(III) and chloramphenicol, an inhibitor of protein synthesis. These results indicate that: (i) the reduction of Fe(III) can occur at low O2 concentrations in acidic sediments; (ii) Fe(II) can be oxidized at O2 concentrations near saturation; and (iii) the enzyme(s) responsible for the reduction of Fe(III) in A. cryptum JF‐5 are not constitutive.
In situ studies of the phylogeny and physiology of filamentous bacteria with attached growth
Authors: Thomsen Trine Rolighed , Kjellerup Birthe Venø , Nielsen Jeppe Lund , Hugenholtz Philip , Nielsen Per Halkjær 
Published: March 2003

SummaryAmong the filamentous bacteria occasionally causing bulking problems in activated sludge treatment plants, three morphotypes with attached microbial growth are common, Eikelboom Type 0041, Type 1851 and Type 1701. A better knowledge of the phylogeny and physiology of these filamentous bacteria is necessary in order to develop control strategies for bulking. In this study we have used a combination of fluorescence in situ hybridization (FISH) and microautoradiography (MAR) to investigate the identity and in situ physiology of the Type 0041‐morphotype and its attached bacteria in two wastewater treatment plants. Identification and enumeration of Type 0041 using group‐specific 16S rRNA‐targeted FISH probes revealed that approximately 15% of the filaments hybridized with a gene probe specific for the TM7 group, a recently recognized major lineage in the bacterial domain. All other filaments morphologically identified as Type 0041 only hybridized to the general bacterial EUB338‐probe, indicating that they probably do not belong to commonly isolated bacterial phyla such as the Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes, for which group‐specific probes were used. The phylogenetic heterogeneity of Type 0041 again highlights the inadequacy of a morphology‐based classification system. Like the filaments, most of the attached microbial cells were not identified beyond their affiliation to the Bacteria using the group‐specific FISH probes. However, several different bacterial phyla were represented in the identified fraction suggesting that the attached microorganisms are phylogenetically diverse. The study of the in situ physiology of Type 0041 using MAR‐FISH revealed that both the filaments and the attached bacteria on Type 0041 were versatile in the use of organic substrates and electron acceptors. It was observed that all Type 0041 could consume glucose, but none of the filaments were able to consume acetate under any conditions tested, in contrast to some of the attached bacteria. No significant physiological differences were found between TM7–positive and TM7–negative Type 0041 filaments, and only minor differences were observed between the two treatment plants tested. These are the first data on the physiology of the almost entirely uncharacterized TM7 phylum and show that TM7 filamentous bacteria can uptake carbon substrates under aerobic and anaerobic conditions.
Multiple influences of nitrate on uranium solubility during bioremediation of uranium‐contaminated subsurface sediments
Authors: Finneran Kevin T. , Housewright Meghan E. , Lovley Derek R. 
Published: March 2003

SummaryMicrobiological reduction of soluble U(VI) to insoluble U(IV) has been proposed as a remediation strategy for uranium‐contaminated groundwater. Nitrate is a common co‐contaminant with uranium. Nitrate inhibited U(VI) reduction in acetate‐amended aquifer sediments collected from a uranium‐contaminated site in New Mexico. Once nitrate was depleted, both U(VI) and Fe(III) were reduced concurrently. When nitrate was added to sediments in which U(VI) had been reduced, U(VI) reappeared in solution. Parallel studies with the dissimilatory Fe(III)‐, U(VI)‐ and nitrate‐reducing microorganism, Geobacter metallireducens, demonstrated that nitrate inhibited reduction of Fe(III) and U(VI) in cell suspensions of cells that had been grown with nitrate as the electron acceptor, but not in Fe(III)‐grown cells. Suspensions of nitrate‐grown G. metallireducens oxidized Fe(II) and U(IV) with nitrate as the electron acceptor. U(IV) oxidation was accelerated when Fe(II) was also added, presumably due to the Fe(III) being formed abiotically oxidizing U(IV). These studies demonstrate that although the presence of nitrate is not likely to be an impediment to the bioremediation of uranium contamination with microbial U(VI) reduction, it is necessary to reduce nitrate before U(VI) can be reduced. These results also suggest that anaerobic oxidation of U(IV) to U(VI) with nitrate serving as the electron acceptor may provide a novel strategy for solubilizing and extracting microbial U(IV) precipitates from the subsurface.
Quality of life of patients after surgical treatment of anal fistula; the role of anal manometry
Authors: Mylonakis E. , Katsios C. , Godevenos D. , Nousias B. , Kappas A. M. 
Published: March 2003

ObjectiveThis study was undertaken to assess the quality of life of patients after surgical treatment of anal fistula and to investigate whether anal manometry (AM) can guide the choice of the proper surgical intervention in these patients in order to protect the sphincter mechanism.Patients and methodsOne hundred patients with anal fistula (AF) were studied prospectively (78 men; mean age 45 years; range 11–78). Cleveland Incontinence Score (CIS) was record pre‐operatively and 1 and 3 months postoperatively for each patient in order to specify their quality of life (QOL) before and after the surgical treatment. Also, anal manometry (AM) was performed pre‐operatively and 1 month postoperatively. The pre‐operative anal pressures and the type of fistula determined the kind of the surgical treatment. 55 patients had an intersphincteric fistula, 42 trans‐sphincteric and 3 suprasphincteric. 65 patients underwent laying open of the fistulous track, 7 fistulectomy and 28 were treated by seton fistulotomy.ResultsThree patients had defective gas control and 6 reported some degree of soiling. 3 patients developed recurrent fistula. CIS was significantly impaired (P=0.02) at the first postoperative month in these patients who were treated for trans‐sphincteric fistula by fistulotomy; AM revealed significant decrease of anal pressures in these patients (resting and squeeze; P=0.007 and 0.0001 respectively); CIS and AM in the remaining cases revealed no significant deterioration of QOL and fall of anal pressures respectively. CIS was normal in the vast majority of patients at 3‐months postoperatively.ConclusionsQOL of patients after surgical treatment of AF is unalterable on the understanding that the AF is simple and the treatment is not associated by incontinence or recurrence. Pre‐operative AM is important regarding the choice of the proper surgical procedure.
Accuracy of hydrogen peroxide‐enhanced ultrasound in the identification of internal openings of anal fistulas
Authors: Ortiz H. , Marzo J. , Jiménez G. , DeMiguel M. 
Published: March 2003

AbstractObjective  This prospective study was designed to assess the accuracy of hydrogen peroxide‐enhanced ultrasound in the identification of internal openings of anal fistulas, with surgical findings as the golden standard.Patients and methods  A total of 143 consecutive patients (102 men; mean age, 45 years) with fistula‐in‐ano were assessed by hydrogen peroxide‐enhanced ultrasound before surgery involving one radiologist. Ultrasound was performed using a B & K Diagnostic Ultrasound System™ with a 10‐MHz rotating endoprobe. Hydrogen peroxide (3%) was infused into the fistula. All operations were perfomed by the same surgeon who was unaware of results of anal endosonography.Results  In 128 (89.5%) patients, an internal opening was identified at surgery. Correct identification of an internal opening endosonographically was recorded in 80 (62.5%) patients. The internal opening was correctly identified by ultrasound in 32% (8/25) of patients with intersphincteric fistulas, in 77% (70/91) with transsphincteric fistulas, and in 17% (2/12) with suprasphincteric fistulas.Conclusion  The accuracy of hydrogen peroxide‐enhanced anal endosonography for the identification of internal openings was still insufficient to justify pre‐operative endosonography as a diagnostic method for routine use in patients with fistula‐in‐ano.
Early insulin release effectively improves glucose tolerance: studies in two rodent models of type 2 diabetes mellitus
Authors: De Souza C. J. , Gagen K. , Chen W. , Dragonas N. 
Published: March 2003

SummaryAim  Islet dysfunction, characterized by the loss of an acute insulin secretory response (AIR) to glucose is a well‐established pathology of type 2 diabetes mellitus. Using oral insulin secreting agents with very different pharmacodynamic profiles, the present study was undertaken to test the hypothesis that, within the setting of an underlying insulin resistance, changes in the insulin response profile can differentially affect glycaemic control.Method  The mildly insulin resistant high‐fat fed Sprague Dawley (HF) rat and the very insulin resistant Zucker fatty (fa/fa) rat, chronically fitted with indwelling jugular cannula were subjected to an oral glucose load. Compounds were administered 5 min before the oral glucose load.0Nateglinide (Nateg) was administered to elicit only an early insulin secretory response and glipizide (Glip) to elicit a later but greater insulin secretory response. Acetaminophen was used as a marker to assess for potential effects of these compounds on gastric emptying rates.Results  Nateg rapidly increased early insulin release (from −5 to 0) while the effects on total insulin release were similar to those in the controls and glucose excursions were eliminated in both diabetic models with no evidence of sustained hypoglycaemia. Conversely, Glip did not affect early insulin release but increased total insulin release (− 15 to 120 min), but only after the oral glucose load. Glip partially curbed glucose excursions in the mildly insulin resistant HF rodent and was totally ineffective in the very insulin resistant Zucker rat. The differential effects could not be attributed to effects on gastric emptying rates.Conclusion  These data support the importance of early insulin release in type 2 diabetes mellitus and indicate that, independent of the level of insulin resistance, stimulating insulin release early and briefly provides for more effective and tighter glycaemic control than increasing insulin exposure to a greater magnitude later.
Brain‐derived neurotrophic factor ameliorates lipid metabolism in diabetic mice
Authors: Tsuchida A. , Nonomura T. , Nakagawa T. , Itakura Y. , Ono‐Kishino M. , Yamanaka M. , Sugaru E. , Taiji M. , Noguchi H. 
Published: March 2003

Aim: It has been reported previously that brain‐derived neurotrophic factor (BDNF) regulates blood glucose metabolism in rodent obese diabetic models such as C57BL/KsJ‐leprdb/leprdb (db/db) mice. BDNF further regulates energy expenditure, possibly through the central and autonomous nervous systems. In this study, we evaluated the effect of BDNF on both lipid and glucose metabolisms to clarify its action mechanism.Methods: To control the energy intake, we used a pellet pair‐feeding apparatus to synchronize food intake precisely between BDNF‐treated and vehicle‐treated db/db mice. BDNF (50 mg/kg/week) was subcutaneously injected to male db/db mice twice weekly for 3 weeks, and blood glucose, serum biochemical lipid parameters and tissue weights were measured. Liver triglyceride contents were measured and liver sections were histologically analysed.Results: Twice weekly BDNF treatment for 3 weeks significantly lowered blood glucose compared with pellet pair‐fed, vehicle‐treated db/db mice (294 ± 109 vs. 529 ± 91 mg/dL). Serum non‐esterified free fatty acid (726 ± 72 vs. 999 ± 220 µEq/l), total cholesterol (125 ± 8 vs. 151 ± 23 mg/dL) and phospholipid levels (215 ± 13 vs. 257 ± 36 mg/dL) of the BDNF‐treated db/db mice decreased significantly. Liver weights (1.51 ± 0.11 vs. 2.05 ± 0.11 g), liver triglyceride contents (17.5 ± 1.4 vs. 26.1 ± 2.1 mg/g) and fatty liver in histological appearance were reduced with BDNF treatment. There were no significant differences in body weights and white adipose tissue weights between the two groups.Conclusions: Taken together with the accelerating effect of BDNF on energy metabolism, these findings indicate that BDNF improves glucose and lipid metabolism in obese diabetic animals without enlarging liver or adipose tissues.
Socio‐demographic and psychosocial factors are associated with features of the metabolic syndrome. The Women's Health in the Lund Area (WHILA) study
Authors: Lidfeldt J. , Nyberg P. , Nerbrand C. , Samsioe G. , Scherstén B. , Agardh C. D. 
Published: March 2003

Aim: The aim was to analyse any associations between socio‐demographic and psychosocial factors and different features of the metabolic syndrome in a geographically well‐defined population of middle‐aged women.Methods: A population of 10 766 Caucasian women aged 50–59 years was investigated regarding biological and socio‐demographic conditions, physical activity, dietary habits, aspects of quality of life, and subjective physical and mental symptoms. The screening instrument was used to discriminate subjects as positive or negative on one or more of a total of eight variables considered to be linked to the metabolic syndrome. The cut‐off values for positive screening were non‐fasting capillary blood glucose ≥ 8.0 mmol/l and serum triglycerides ≥ 2.3 mmo/l, BMI ≥ 30 kg/m2, WHR ≥ 0.90, blood pressure ≥ 160 and/or 95 mmHg, a family history of diabetes, and pharmacological treatment for hypertension or hyperlipidaemia.Results: Altogether 6805 women (63.2%) participated: 3535 with positive and 3270 with negative screening. Multiple logistic regression analyses showed that comprehensive (OR 1.62, 95% CI 1.41–1.87) and upper secondary (1.40, 1.24–1.57) school, low physical quality of life (1.41, 1.23–1.61) and high sum of subjective physical symptoms (1.06, 1.04–1.08) were positively associated with one or more features of the metabolic syndrome, while high leisure‐time exercise and healthy diet (0.84, 0.71–0.99), and low (≤ 83 g/week) (0.71, 0.63–0.81) and moderate (84–167 g/week) (0.78, 0.65–0.93) alcohol consumption were negatively associated.Conclusions: To identify middle‐aged women with cardiovascular risk factors and high risk for diabetes, it is important to consider not only biological, but also socio‐demographic and psychosocial conditions.
Relationship between serum ferritin levels and tumour status in patients with renal cell carcinoma
Authors: Miyata Y. , Koga S. , Nishikido M. , Hayashi T. , Kanetake H. 
Published: March 2003

Objective To assess the relationship between serum ferritin levels (a useful marker for diagnosing and staging renal cell carcinoma, RCC) and tumour status in RCC of  7 cm diameter. Patients and methods Serum ferritin levels were measured using an enzyme immunoassay in 101 patients before treatment for RCC. The largest diameter of the primary tumour was measured in the pathological specimens. The correlation between serum ferritin levels and histopathological status was assessed retrospectively in all patients, and separately in the 67 patients with tumours of  7 cm diameter. Results For the whole group, lymph node metastasis, distant metastasis, extracapsular invasion, vascular invasion and nuclear grade were significantly different (P < 0.001) between patients with normal and those with elevated ferritin levels. However, in patients with tumours of  7 cm, only the presence of distant metastasis was significantly different (P < 0.001) between the groups. Conclusions Elevated serum ferritin levels in patients with RCC of  7 cm may suggest the presence of distant metastasis.