bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cloning and disruption of the Pichia pastoris ARG1, ARG2, ARG3, HIS1, HIS2, HIS5, HIS6 genes and their use as auxotrophic markers
Authors: Nett Juergen H. , Hodel Nikolai , Rausch Sebastian , Wildt Stefan 
Published: March 2005

AbstractScreening of a partial genomic database of Pichia pastoris allowed us to identify the ARG1, ARG2, ARG3, HIS1, HIS2, HIS5 and HIS6 genes, based on homology to their Saccharomyces cerevisiae counterparts. Based on the cloned sequences, a set of disruption vectors was constructed, using the previously described PpURA5‐blaster as a selectable marker, and the cloned genes were individually disrupted. All disruptants exhibited the expected auxotrophic phenotypes, with only the his2 knockouts displaying a bradytroph phenotype. To allow their use as auxotrophic markers, we amplified the open reading frames and respective promoters and terminator regions of PpARG1, PpARG2, PpARG3, PpHIS1, PpHIS2 and PpHIS5. We then designed a set of integration vectors harbouring cassettes of the ARG pathway as selectable markers, to disrupt the genes of the HIS pathway and vice versa. Employing this strategy, we devised a scheme allowing for the rapid and stable introduction of several heterologous genes into the genome of P. pastoris without the need for recyclable markers or strains with multiple auxotrophies. Furthermore, simple replica‐plating, instead of cost‐consuming and labour‐intensive colony PCR or Southern analysis, can be used to identify positive transformants, making this approach amendable for initial high‐throughput applications, which can then be followed up by a more careful analysis of the selected transformants. The sequences presented here have been submitted to the EMBL data library under Accession Nos/AY532165 (ARG1), AY532166 (ARG2), AY532167 (ARG3), AY532168 (HIS1), AY532169 (HIS2), AY532170 (HIS5), AY532171 (HIS6). Copyright © 2005 John Wiley & Sons, Ltd.
Expression of GCR1, the transcriptional activator of glycolytic enzyme genes in the yeast Saccharomyces cerevisiae, is positively autoregulated by Gcr1p
Authors: Sasaki Hiromi , Kishimoto Tomoko , Mizuno Takayuki , Shinzato Tomoko , Uemura Hiroshi 
Published: March 2005

AbstractWhen regulation of GCR1 expression was analysed using a GCR1–lacZ fusion, lacZ expression levels were decreased in the Δgcr1 or Δgcr2 mutant. RT–PCR analysis of genomic GCR1 transcript confirmed the dependency of GCR1 expression on the Gcr1p–Gcr2p complex. Examination of the 5′ non‐coding region of GCR1 identified three putative Gcr1p binding sites (CT‐boxes) in the −100 to −200 region of GCR1, and the putative binding sites for Rap1p (RPG‐box) and Abf1p were also identified nearby. The region containing putative cis‐elements was analysed by cloning it upstream of the CYC1TATA–lacZ fusion. The GCR1UAS–CYC1TATA–lacZ fusion showed a moderate activity and, as expected, the activity was drastically reduced in the Δgcr1 or Δgcr2 mutant. Systematic deletion and mutation analyses of cis‐elements in this region demonstrated that the putative binding sites for Rap1p and Abf1p were not involved in the promoter activity of GCR1UAS and only one of the three CT‐boxes showed GCR1‐ and GCR2‐dependent promoter activity. In contrast to the expression of glycolytic genes, where a RPG‐box adjacent to the CT‐box is required for strong promoter activities, CT‐box‐dependent expression of GCR1 did not require the RPG‐box. Also, a contribution of Sgc1p, an E‐box binding transcription factor, to the expression of GCR1 was suggested, based on its disruption analysis. Copyright © 2005 John Wiley & Sons, Ltd.
Surgically induced uremia in rats I: Effect on bone strength and metabolism
Authors: Jablonski G. , Klem K. H. , Attramadal A. , Dahl E. , Rønningen H. , Gautvik K. M. , Haug E. , Gordeladze J. O. 
Published: March 2005

During the course of chronic renal failure (CRF) in man, renal osteodystrophy (osteitis fibrosa and/or osteomalacia) gradually develops. The present study aimed to establish a similar type of CRF leading to renal osteodystrophy in rats.During progressive CRF development over 225 days after 5/6 nephrectomy, the following serum variables were measured: creatinine, immunoreactive parathryoid hormone (iPTH), 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), a25-hydroxyvitamin D3, (25(OH)D3), alkaline phosphatase, albumin, phosphate, urea nitrogen, total calcium, and other blood electrolytes. Subsequent to sacrifice, mechanical properties of the rat femur, bone histomorphometry (osteoid and eroded surfaces) and bone contents of calcium, phosphate and hydroxyproline were also examined.Serum creatinine in rats with CRF gradually escalated by some 70%, while circulating 1,25(OH)2D3 was reduced beneath detection level. Total plasma calcium and phosphate concentrations were, however, almost unchanged indicating that PTH-induced bone remodeling due to moderate hyperparathyroidism sustained calcium homeostasis. Alkaline phosphatase levels were reduced by some 50%, which reflects chronically impeded bone formation. Bone histomorphometry assessment revealed substantial elevation of resorption with moderate accompanying fibrosis in about 70% of afflicted animals. Bone calcium, phosphate and hydroxpyroline contents remained unaltered. However, hydroxoproline/calcium ratio was marginally reduced. These results, together with altered mechanical bending stress characteristics and diminished diaphysis cross section area, confirm development of mixed bone lesions in the uremic animals.Our results are compatible with the early development of CRF in man. The established rat model is therefore useful in elucidating the precipitation and early treatment of renal osteodystrophy in humans.
Impact of sleep‐related breathing disorders on health‐related quality of life in patients with chronic heart failure
Authors: Skobel Erik , Norra Christine , Sinha Anil , Breuer Christian , Hanrath Peter , Stellbrink Christoph 
Published: March 2005

BackgroundQuality of life in patients with chronic heart failure (HF) is often severely compromised. Sleep‐related breathing disorders (SRBD) like Cheyne–Stokes Respiration (CSR) or obstructive sleep apnea (OSAS) are often observed in patients with severe HF resulting in fragmentation of sleep, excessive daytime sleepiness and an increased mortality. While an apnea/hypopnea‐index (AHI) >30/h represents an independent predictor of poor prognosis, clinical relevance of even minor SRBD with an AHI <30/h remains unclear with respect to quality of life, exercise capacity or depression rate.MethodsSixty‐nine consecutive ambulatory patients with stable HF (NYHA II–III, EF 25%) underwent two night polygraphies with a six‐channel ambulatory recording. Spiroergometry was performed, and patients were examined for sleep quality (PSQI), depressed mood (BDI) and health‐related quality of life (SF‐36). The data were compared to 10 age‐matched healthy controls and 11 patients with OSAS (AHI 14–29/h) not suffering from HF.ResultsFifty‐one patients completed follow up. 52% were positively diagnosed for SRBD (AHI 16–30/h: 12 patients CSR, 5 patients OSAS, 9 patients mixed); 25 patients (48%) showed no relevant SRBD. Patients with HF and SRBD had lower quality of life than patients without SRBD and HF. The severity of SRBD as indicated by the AHI significantly correlated with quality of life measures: Bodily pain, physical functioning and social functioning showed largest impairment in patients with HF and SRBD. Furthermore, elevated depression rates in correlation to the AHI were only observed in patients with SRBD similar to patients with OSAS without HF.ConclusionEven minor SRBD in patients with HF independently influence quality of life and correlate with estimation of depression and sleep disturbances.
The additive prognostic value of restrictive pattern and dipyridamole‐induced contractile reserve in idiopathic dilated cardiomyopathy
Authors: Pratali Lorenza , Otasevic Petar , Rigo Fausto , Gherardi Sonia , Neskovic Aleksandar , Picano Eugenio 
Published: March 2005

AbstractBackgroundDiastolic dysfunction and lack of contractile reserve are unfavorable prognostic predictors in patients with dilated cardiomyopathy (DCM).AimsThis study aims to assess whether diastolic dysfunction and lack of dipyridamole‐induced contractile reserve were additive predictors of poor outcome in patients with DCM.MethodsA total of 116 patients with DCM and ejection fraction (EF<35%) were studied by dipyridamole echo (0.84 mg/kg over 10 min). At rest, a restrictive filling pattern was defined as: E/A ratio >2 and an E‐wave deceleration time of <140 ms on transmitral flow velocity profile.ResultsRest wall motion score index (WMSI) was 2.2±0.3 and decreased to 1.9±0.41 after dipyridamole (p<0.001). During follow‐up (median 26.5 months), 22 cardiac deaths occurred. At multivariate analysis, dipyridamole‐induced contractile reserve yielded significant incremental prognostic value (RR=0.275, p<0.006) over NYHA class (RR=1.971, p<0.03), angiotensin‐converting enzyme inhibitor therapy (RR=0.173, p<0.001), and left ventricular end‐diastolic diameter (RR=1.131, p<0.001). The worst prognostic combination was the presence of restrictive pattern at rest and the absence of contractile reserve (ΔWMSI<0.15).ConclusionIn patients with DCM, the ominous combination of restrictive transmitral flow pattern and lack of contractile reserve during dipyridamole stress predicts an unfavourable outcome.
Lévy trajectories of Peruvian purse-seiners as an indicator of the spatial distribution of anchovy (Engraulis ringens)
Authors: Bertrand Sophie , Burgos Julian M. , Gerlotto François , Atiquipa Jaime 
Published: March 2005

Abstract
Analogous to other top predators foraging on patchy resources, the spatial behaviour of fishers provides information on the spatial organization of fish. Focusing on the Peruvian anchovy purse-seine fishery, vessel monitoring system data are used to describe fishing vessels' trajectories, and acoustic survey data to characterize anchovy spatial distribution. Vessel trajectories were analysed in terms of move-length distribution, whereas fish distribution was characterized by spatial extent, concentration, and fractal dimension. Fishers perform Lévy flights that can be characterized by a single statistic (μ); μ is significantly correlated with the fractal dimension of fish distribution. It is argued that the Lévy statistic is a good candidate for an ecosystem indicator that might contribute to real-time monitoring of ecosystems.
An extremal function for the Chang-Marshall inequality over the Beurling functions
Authors: Andreev Valentin 
Published: March 2005

S.-Y. A. Chang and D. E. Marshall showed that the functional

Λ
(
f
)
=
(
1

/

2
π
)

∫
0

2
π

exp
⁡
{

|

f
(

e

i
θ

)

|

2

}
d
θ

\Lambda (f) =(1/2\pi ) \int _0^{2\pi }\exp \{ |f(e^{i\theta })|^2\}d\theta

is bounded on the unit ball

B

\mathcal {B}

of the space

D

\mathcal {D}

of analytic functions in the unit disk with

f
(
0
)
=
0

f(0)=0

and Dirichlet integral not exceeding one. Andreev and Matheson conjectured that the identity function

f
(
z
)
=
z

f(z)=z

is a global maximum on

B

\mathcal {B}

for the functional

Λ
\Lambda

. We prove that

Λ
\Lambda

attains its maximum at

f
(
z
)
=
z

f(z)=z

over a subset of

B

\mathcal {B}

determined by kernel functions, which provides a positive answer to a conjecture of Cima and Matheson.
German Lawyers and Globalisation: Changing Professional Identity
Authors: Shaw Gisela 
Published: March 2005

abstractThe emergence of modern professions on the continent of Europe has generally been associated with the emergence of nation states. This is particularly true of the legal professions which, by their very nature, are characterised by a special interest in and relationship with the state. The late arrival on the scene of a German nation state meant that the process of professionalisation also suffered relative delay. No truly unified liberal professions could emerge before there was a national government with which to negotiate their roles.Today, harmonisation initiatives of the European Union, embedded in and reinforced by growing globalisation of world markets, have set in motion a process of unprecedented professional de‐ and/or re‐regulation, curtailing nation state control and exposing professions to increasing internal and external competition at national and international levels. This paper looks at the impact of these processes on the ‘Rechtsanwaltschaft’, a key player in the German legal services market. Current developments are assessed in the light of a tradition of strict state regulation coupled with a high level of state‐protected privileges. The conceptual framework draws on research from the sociology of the professions.
6‐Tioguanine monitoring in steroid‐dependent patients with inflammatory bowel diseases receiving azathioprine
Authors: Roblin X. , Serre‐Debeauvais F. , Phelip J.‐M. , Faucheron J.‐L. , Hardy G. , Chartier A. , Helluwaert F. , Bessard G. , Bonaz B. 
Published: March 2005

SummaryBackground : 6‐Thioguanine (6‐tioguanine) nucleotides are the active metabolites of azathioprine.Aim : The aim of the study was to evaluate the rate of clinical remission without steroids in steroid‐dependent Crohn's disease and ulcerative colitis patients receiving azathioprine, the medium‐ and long‐term efficacy and the predictive factors of clinical response when monitoring 6‐tioguanine.Methods : Steroid‐dependent Crohn's disease and ulcerative colitis patients receiving either azathioprine or not (treated later with a daily dose of 2.5 mg/kg) were prospectively included. 6‐Tioguanine was monitored at 1 and 2 months and every 3 months thereafter for 1 year. The azathioprine dose was adapted to reach a 6‐tioguanine level of >250 pmol/8 × 108 red blood cells. Thiopurine methyltransferase genotype/phenotype was evaluated in some patients.Results : A total of 106 patients were prospectively included (70 Crohn's disease, 36 ulcerative colitis). The clinical remission rate without steroids in patients receiving azathioprine, in intention‐to‐treat analysis, was 72% and 59% at 6 and 12 months, respectively. The remission rate was significantly higher in patients with 6‐tioguanine >250 pmol/8 × 108 RBC (86% and 69% at 6 and 12 months, respectively; P < 0.01). No significant difference was observed between Crohn's disease and ulcerative colitis patients whether treated by azathioprine or not on inclusion. In the univariate analysis, the absence of Crohn's disease stenosis, a 6‐tioguanine level >250 pmol/8 × 108 RBC, and an increase of erythrocyte mean corpuscular volume were the factors predictive of a favourable clinical response. In the multivariate analysis, only a 6‐tioguanine level of >250 pmol/8 × 108 red blood cells was a predictive factor of favourable clinical remission.Conclusions : Clinical remission without steroids is significantly more likely when monitoring 6‐tioguanine so as to reach a level of >250 pmol/8 × 108 red blood cells in steroid‐dependent Crohn's disease and ulcerative colitis patients receiving azathioprine (86% and 69% at 6 and 12 months, respectively).
The out‐patient management of patients with acute mild‐to‐moderate colonic diverticulitis
Authors: Mizuki A. , Nagata H. , Tatemichi M. , Kaneda S. , Tsukada N. , Ishii H. , Hibi T. 
Published: March 2005

SummaryBackground : There are no management criteria for optimum out‐patient care in mild‐to‐moderate acute colonic diverticulitis.Aim : To enable such patients to be managed in an out‐patient setting, by establishing criteria and treatment protocols.Methods : We conducted an open trial and follow‐up study from 1997 to 2002. On the basis of ultrasonography, we defined and categorized mild‐to‐moderate acute colonic diverticulitis ranging from limited inflammation within diverticulum to an abscess < 2 cm in diameter. Subjects were treated as out‐patients and followed a 10‐day treatment protocol consisting of an oral antibiotic and a sports drink for the first 3 days. Physical examination and laboratory testing helped determine whether or not a patient could resume a liquid diet on day 4, and a regular diet on day 7.Results : Of the 70 patients, 68 were successfully treated. Two patients required hospitalization. Of the 65 patients who were tracked over several months [median (intraquarter range) = 30.8 (11.9–44.2) months], 16 had one or more clinical recurrences. The medical cost per episode was 80% lower than in‐patient treatment.Conclusions : Patients with mild‐to‐moderate acute colonic diverticulitis can be safely and successfully treated as out‐patients using this protocol.
Helicobacter pylori duodenal colonization is a strong risk factor for the development of duodenal ulcer
Authors: Pietroiusti A. , Luzzi I. , Gomez M. J. , Magrini A. , Bergamaschi A. , Forlini A. , Galante A. 
Published: March 2005

SummaryAim : To test the hypothesis that duodenal colonization represents the final crucial step in the development of Helicobacter pylori related duodenal ulcer.Methods : Patients with non‐ulcer dyspepsia who had gastric colonization by H. pylori were included in the study. At baseline endoscopy, we evaluated the prevalence of duodenal colonization (culture, urease testing and histology), and cytotoxin‐associated gene A status (polymerase chain reaction). No patients received eradication during 1 year follow‐up. At this time, endoscopy was repeated and the incidence of duodenal ulcer was assessed.Results : Among 181 patients completing follow‐up, 53 (29%) had duodenal colonization: 72% of them were cytotoxin‐associated gene A positive, versus 37% patients without duodenal colonization (P < 0.001). Duodenal ulcer developed in 12 (22.6%) patients with duodenal colonization and in two (1.6%) without duodenal colonization (odds ratio for duodenal ulcer: 6.29, 95% confidence intervals 2.44–17.45). The incidence of duodenal ulcer was similar among cytotoxin‐associated gene A positive and cytotoxin‐associated gene A negative subjects with duodenal colonization: 21.05% versus 26.6%.Conclusions : The assessment of duodenal colonization by H. pylori in patients with non‐ulcer dyspepsia is strongly predictive for the subsequent development of duodenal ulcer and may help to stratify patients at risk for this disease.
Genotypes of the pancreatic β‐cell K‐ATP channel and clinical phenotypes of Japanese patients with persistent hyperinsulinaemic hypoglycaemia of infancy
Authors: Ohkubo Kumiko , Nagashima Mariko , Naito Yuko , Taguchi Tomoaki , Suita Sachiyo , Okamoto Nobuhiko , Fujinaga Hideshi , Tsumura Kumi , Kikuchi Kiyoshi , Ono Junko 
Published: March 2005

SummaryObjective  Persistent hyperinsulinaemic hypoglycaemia of infancy (PHHI) is a disorder of glucose metabolism that is characterized by dysregulated secretion of insulin from pancreatic β‐cells. This disease has been reported to be associated with mutations of the sulfonylurea receptor SUR1 (ABCC8) or the inward‐rectifying potassium channel Kir6.2 (KCNJ11), which are two subunits of the pancreatic β‐cell ATP‐sensitive potassium channel.Patients and Methods  In 14 Japanese PHHI patients, all exons of SUR1 and Kir6.2 genes were analysed by polymerase chain reaction (PCR) and direct sequencing. Four patients responded to diazoxide, and nine patients underwent a subtotal pancreatectomy. Histologically, seven patients were diagnosed to have a focal form and two a diffuse form of the disease.Results  We found nine novel mutations in the SUR1 gene and two in the Kir6.2 gene. In the SUR1 gene mutations, three were nonsense mutations (Y512X, Y1354X and G1469X), one was a one‐base deletion in exon 7, and two were missense mutations in the nucleotide‐binding domain 2 (K1385Q, R1487K). The other three mutations occurred in introns 14, 29 and 36, which might cause aberrant splicing of RNA. Two siblings in one family were heterozygotes for a missense mutation, K1385Q, which was maternally inherited. In Kir6.2 gene screening, one patient was found to be a compound heterozygote of a missense mutation (R34H) and a one‐base deletion (C344fs/ter).Conclusion  The novel mutations reported here could be pathological candidates for PHHI in Japan. They also reveal that SUR1 and Kir6.2 mutations in the Japanese population exhibit heterogeneity and that they occurred at a frequency similar to other genetic populations.
Prevalence and progression of subclinical hypothyroidism in women with type 2 diabetes: the Fremantle Diabetes Study
Authors: Chubb S. A. P. , Davis W. A. , Inman Z. , Davis T. M. E. 
Published: March 2005

SummaryObjective  To assess the prevalence and progression of subclinical hypothyroidism in women with type 2 diabetes.Design and Patients  Cross‐sectional and longitudinal observational assessment of thyroid function in 420 adult females with type 2 diabetes randomly selected from participants in the community‐based Fremantle Diabetes Study.Measurements  Serum TSH, antibodies to thyroperoxidase (anti‐TPO) and serum free T4 were measured at baseline and after 5 years. Baseline glycated haemoglobin (HbA1c), serum glucose, serum total and high density lipoprotein (HDL) cholesterol, serum triglycerides and antibodies to glutamic acid decarboxylase (anti‐GAD) were also used in analyses.Results  After exclusion of patients with known thyroid disease or taking amiodarone or lithium at baseline, the prevalence of subclinical hypothyroidism (a raised serum TSH and normal serum free T4) was 8·6%. Subclinical hypothyroidism was associated with anti‐TPO status and age, but there were no independent associations with serum cholesterol, history of coronary heart disease, HbA1c or hypoglycaemic therapy. In the subgroup of patients restudied after 5 years, none of those who had subclinical hypothyroidism at baseline had overt hypothyroidism regardless of anti‐TPO status.Conclusions  In women with type 2 diabetes without known thyroid disease, subclinical hypothyroidism is a common but incidental finding. The routine screening of thyroid function in type 2 diabetes is questionable.
Beyond the regeneration phase: differentiation of height–light trajectories among tropical tree species
Authors: POORTER LOURENS , BONGERS FRANS , STERCK FRANK J. , WÖLL HANNSJÖRG 
Published: March 2005

Summary

A height–light trajectory (HLT, a fitted curve relating canopy exposure to tree height) was determined for populations of individuals of each of 47 tree species in a Liberian lowland rainforest. The HLTs were compared and related to tree allometry and adult stature. Crown exposure was measured for 7460 trees and related to tree height using a multinomial regression analysis. Individual trees were followed for 2.8–9.8 years.
The trajectories of the 47 species were compared with the average vertical light profile in the forest canopy. Evidence was found for the existence of all nine trajectories hypothesized on the basis of three possible light environments (high, intermediate and low) for juveniles and adults. The classical paradigm of pioneer vs. shade tolerant, based on seed and seedling responses, does not therefore apply to post‐seedling stages.
The majority of the species followed the vertical light profile in the forest canopy, starting in low light environments in the juvenile stage and ending up in high light environments in the adult stage. Only two species complied with the classic notion of whole‐life shade tolerants and whole‐life shade intolerants (one each).
The predictable vertical light gradient in the forest canopy has led to a close association between adult height, light trajectories and allometric traits. Large‐stature species tend to have relatively slender stems and narrow crowns, and therefore realize a faster temporal and height‐related increase in crown exposure.
Tree species have different height–light trajectories when they grow from seedling to adulthood. This may have profound repercussions for our current views on plasticity and adaptation, light partitioning and species coexistence, and on silviculture and management.