bluuebunny/crossref_metadata_2025 huggingface dataset:
Genital Automatisms: A Video‐EEG Study in Patients with Medically Refractory Seizures
Authors: Dobesberger Judith , Walser Gerald , Unterberger Iris , Embacher Norbert , Luef Gerhard , Bauer Gerhard , Benke Thomas , Bartha Lisa , Ulmer Hanno , Ortler Martin , Trinka Eugen
Published: July 2004

Summary: Purpose:Genital automatisms (GAs) are rare clinical phenomena during or after epileptic seizures. They are defined as repeated fondling, grabbing, or scratching of the genitals. The anatomic correlates of GAs have been discussed controversially. The aim of this investigation was to assess the localizing and lateralizing value of GAs.

Methods:The authors studied 207 consecutive patients with intractable seizures referred to a University Hospital for presurgical evaluation between 1998 and 2002: 135 had temporal lobe epilepsy (TLE); 23, frontal lobe epilepsy (FLE); 29, generalized epilepsies (GEs); and 20 had extratemporal or multifocal epilepsy.

Results:Twenty‐three (11%) of 207 patients showed GAs in 42 (3%) of 1,299 seizures. GAs occurred significantly more often in men (17 of 93, 18%) than in women (six of 114, 5%; p = 0.0037). Twenty‐one (16%) of 135 patients with TLE performed GAs, one (4%) of 23 with FLE and one (3%) of 29 with GE. GAs were associated with unilateral hand automatisms in 16 (70%) of 23 and with periictal urinary urge in five (22%) of 23. All patients had amnesia for the performance of GAs.

Conclusions:GAs appear in the ictal or postictal period with impaired consciousness. Men exhibit GAs significantly more often than do women. GAs do not localize or lateralize per se, but may localize seizure onset in the presence of periictal urinary urge or unilateral hand automatisms. They show a tendency to occur more often in TLE.
Temperate Utah chub form valid otolith annuli in the absence of fluctuating water temperature
Authors: Johnson J. B. , Belk M. C.
Published: July 2004

Utah chub Gila atraria, a temperate freshwater minnow, formed valid otolith annuli (annual growth rings), even when raised in a constant‐temperature desert spring environment. This suggests that factors other than seasonal variation in water temperature control annual otolith marking.
The use of pulse‐echo acoustic microscopy to non‐invasively determine the sex of living larval sea lampreys
Authors: Maeva E. , Bruno I. , Zielinski B. S. , Docker M. F. , Severin F. M. , Maev R. G.
Published: July 2004

A rapid, accurate and non‐invasive method to determine the sex of larval sea lampreys Petromyzon marinus, using wide‐field pulse‐scanning acoustic microscopy, is described. Cross‐sectional pulse‐echo scans were made with a high‐resolution acoustic microscope in 48 larvae (110–130 mm total length, LT), and the acoustic images generated showed such internal body structures as the gonad, intestine, kidneys, cardinal veins, notochord and musculature. Females were identified by the presence of a relatively large (1–1·5 mm diameter) ovary, which was considerably less reflective to the 15 to 25 MHz acoustic signals than the surrounding kidney tissue. Males were recognized by the lack of the large non‐reflective ovary and, in some cases, the appearance of a small (0·2–0·3 mm) testis with slightly stronger reflective properties than the kidney. Identification of sex was confirmed by optical microscopy following dissection, and in a blind test on an additional 10 specimens (121–168 mm LT), the acoustic method reliably identified sex in 100% of the larvae. These results indicate that acoustic microscopy can determine the sex of live sea lamprey larvae in c. 30 s per animal, a process which until now required dissection or invasive surgery.
Sound production during competitive feeding in the grey gurnard
Authors: Amorim M. C. P. , Stratoudakis Y. , Hawkins A. D.
Published: July 2004

The acoustic repertoire of captive grey gurnard Eutrigla gurnardus during competitive feeding consisted of three types of sound: knocks, grunts and growls. Knocks were audible as a single sound, whereas grunts and growls were perceived as longer, pulsed sounds to the human ear. Typically, knocks were composed of 1–2 pulses, grunts of 4–8 pulses and growls >10 pulses. Growls were longer and had shorter pulse periods than grunts. All sound types had peak frequencies of c. 500 Hz. The sequences of behaviours observed during feeding interactions suggest that grey gurnard obtain food both by scramble and contest tactics. Competing fish emitted knocks mainly while grasping a food item and also during other non‐agonistic behaviour, suggesting that knock production may reflect a state of feeding arousal but could also serve as a warning of the forager's presence to nearby competitors. Grunts were mainly emitted during frontal displays, which were the most frequent behavioural act preceding grasps, suggesting that they may play a role in deterring other fish from gaining access to disputed food items.
Estimation of true dihedral angles and relative grain boundary energies in polycrystals using the disector
Authors: CHHABRA S. , SANGAL S. , MUNGOLE M. N.
Published: July 2004

Relative grain boundary energies can be simply related to true dihedral angles, which are the angles between grain boundary planes meeting at triple edges in polycrystals. Some limited efforts in the measurement of true dihedral angles have used the technique of serial sectioning, which is usually cumbersome and time consuming. In this study the three‐dimensional probe called the ‘disector’ has been employed to evaluate true dihedral angles. This probe, combined with automated image processing, introduces precision as well as efficiency, overcomes the disadvantages of the two‐dimensional probe and is far less tedious and less complicated than serial sectioning. It is shown that the technique is relatively simple and therefore can be applied to obtain a significantly large and accurate statistical sample of true dihedral angles. The application of this method is demonstrated by evaluating the triple junction geometry and the associated relative grain boundary energies in polycrystalline 316L austenitic stainless steel.
Possible common biomarkers from FTIR microspectroscopy of cervical cancer and melanoma
Authors: MORDECHAI S. , SAHU R. K. , HAMMODY Z. , MARK S. , KANTAROVICH K. , GUTERMAN H. , PODSHYVALOV A. , GOLDSTEIN J. , ARGOV S.
Published: July 2004

Detection of malignancy at early stages is crucial in cancer prevention and management. Fourier transform infrared (FTIR) spectroscopy has shown promise as a non‐invasive method with diagnostic potential in cancer detection. Studies were conducted with formalin‐fixed biopsies of melanoma and cervical cancer by FTIR microspectroscopy (FTIR‐MSP) to detect common biomarkers, which occurred in both types of cancer distinguishing them from the respective non‐malignant tissues. Both types of cancer are diagnosed on skin surfaces. The spectra were analysed for changes in levels of biomolecules such as RNA, DNA, phosphates and carbohydrate (glycogen). Whereas carbohydrate levels showed a good diagnostic potential for detection of cervical cancer, this was not the case for melanoma. However, variation of the RNA/DNA ratio as measured at I(1121)/I(1020) showed similar trends between non‐malignant and malignant tissues in both types of cancer. The ratio was higher for malignant tissues in both types of cancer.
Negation of lexical have in conversational English
Authors: Nelson Gerald
Published: July 2004

Abstract:  The lexical verb have may be negated in a very wide variety of ways, e.g. I don’t have (any) money/I haven’t any money/I haven’t got any money/I have no money/I’ve got no money. Previous research has shown that the choice of negation pattern distinguishes very clearly between American English (AmE) and British English (BrE). American speakers show a 90 per cent preference for I have no money, while British speakers show an overall preference for I haven’t got any money, though they also frequently use the other available patterns.

In this paper I examine the negation of lexical have in corpus data from Great Britain, New Zealand, India, Hong Kong, and Singapore. My initial hypothesis is that the three Outer Circle varieties (Hong Kong, India, Singapore) should tend to follow one or other of the two dominant “models,” that is, they should follow either the American preference or the very different British preference. In fact, the results do not support this hypothesis. The Outer Circle varieties are shown to form a very distinct group of their own, in terms of the negation of lexical have.
What do we mean by bias, Englishes, Englishes in testing, and English language proficiency?
Authors: Brown James Dean
Published: July 2004

The authors of “Whose norms? International proficiency tests in English”(Davies, Hamp‐Lyons and Kemp, 2003) raise important issues and suggest interesting avenues for future research. As is usually true when a new line of research emerges, defining terms is fundamental. Four sets of terms seem to me to need clarification here: bias; Englishes(in general); Englishes in testing; and English language proficiency.
Specific Language Impairments in Children
Authors: Joanisse Marc F.
Published: July 2004

Theories of specific language impairment (SLI) in children turn on whether this deficit stems from a grammar-specific impairment or a more general speech-processing deficit. This issue parallels a more general question in cognitive neuroscience concerning the brain bases of linguistic rules. This more general debate frequently focuses on past-tense verbs, specifically, whether regular verbs (bake-baked) are encoded as rules, and whether irregular forms (take-took) are processed differently. Children with SLI have difficulties with past tenses, so SLI could represent an impairment to rules. An alternative theory explains past-tense deficits in SLI as resulting from a phonological deficit. Evidence for this theory has been obtained from connectionist models of past-tense impairments and from behavioral studies of language- and reading-impaired children. The data suggest that SLI is not an impairment to linguistic rules, that past-tense impairments can be explained as resulting from a perceptual deficit, and that a single processing mechanism is ideally suited to account for these children's difficulties.
The Macedonian–Albanian political frontier: the re‐articulation of post‐Yugoslav political identities
Authors: Adamson Kevin , Jović Dejan
Published: July 2004

Abstract. The article examines the re‐articulation of national identity in Macedonia since its independence in 1992. Both ethnic Macedonian and ethnic Albanian political identities have been engaged in a complex process of redefinition. Two ethnic groups had previously been strongly influenced by the Marxist paradigm and its Yugoslav official interpretation. During the 1990s, the elements of the old paradigm were combined with elements of the new – liberal democratic – concepts of nationhood. While some of the concepts developed within the old Yugoslav framework are still in use, the new liberal‐democratic political paradigm finds it difficult to include them into an official discourse on nationhood. At the same time, introduction of the concepts inherent to the liberal‐democratic paradigm has disturbed the fragile balance achieved through the old Yugoslav narrative. In new circumstances, the ethnic Macedonians transformed themselves from the ‘constitutive nation’ to ‘majority’. However, the ethnic Albanians found it more difficult to accept the status of ‘minority’, which was once (in Yugoslav Marxist narrative) considered to be politically incorrect. Thus, they insist on being recognised as a ‘nation’, equal to ethnic Macedonians. In its essence, the conflict in Macedonia is – to a large extent – a conflict between two different concepts of what is Macedonia and who are Macedonians. The questions posed are: is the minority (ethnic Albanians) part of the nation? Could two nations exist peacefully within one state? The article maps out differences between two different discourses on the identity of the new Macedonian state.
Gaeumannomyces graminis, the take‐all fungus and its relatives
Authors: FREEMAN JACQUELINE , WARD ELAINE
Published: July 2004

Take‐all, caused by the fungus Gaeumannomyces graminis var. tritici, is the most important root disease of wheat worldwide. Many years of intensive research, reflected by the large volume of literature on take‐all, has led to a considerable degree of understanding of many aspects of the disease. However, effective and economic control of the disease remains difficult. The application of molecular techniques to study G. graminis and related fungi has resulted in some significant advances, particularly in the development of improved methods for identification and in elucidating the role of the enzyme avenacinase as a pathogenicity determinant in the closely related oat take‐all fungus (G. graminis var. avenae). Some progress in identifying other factors that may be involved in determining host range and pathogenicity has been made, despite the difficulties of performing genetic analyses and the lack of a reliable transformation system.
Feeding cell development by cyst and root‐knot nematodes involves a similar early, local and transient activation of a specific auxin‐inducible promoter element
Authors: KARCZMAREK ANETA , OVERMARS HEIN , HELDER JOHANNES , GOVERSE ASKA
Published: July 2004

To study the role of the phytohormone auxin in nematode feeding cell induction and early development, the transcriptional regulation of the artificial auxin‐responsive promoter element DR5 was monitored in Arabidopsis thaliana roots infected with the cyst nematode Heterodera schachtii or the root‐knot nematode Meloidogyne incognita. For both nematode species, a specific and strong activation of DR5::gusA was observed inside the initial feeding cells at 18 h post inoculation, pointing to an increase in the perceived auxin concentration. This high expression was maintained until 3–5 days post inoculation and subsequently the GUS staining was reduced. Cyst and root‐knot nematodes are distantly related and the feeding sites they induce are highly dissimilar. In this respect, the similarities between the two nematode‐induced DR5 activation patterns in A. thaliana roots are remarkable. A transient and local increase in auxin perception could be due to an accumulation or to an increased sensitivity. Based on previously published data, a local auxin accumulation seems to be the more probable explanation. The observed early and localized increase of the perceived IAA concentration in the initial feeding structure underlines that this phytohormone could be an important clue in feeding cell induction by plant parasitic nematodes.
The Nep1‐like proteins—a growing family of microbial elicitors of plant necrosis
Authors: PEMBERTON CLARE L. , SALMOND GEORGE P. C.
Published: July 2004

A novel family of microbial elicitors of plant necrosis has been identified. Designated Nep1‐like proteins (NLPs), after the first family member isolated, they range from 24 to 26 kDa and are found in a variety of taxonomically unrelated micro‐organisms. These include several fungi and oomycetes, as well as Gram‐positive and Gram‐negative bacteria. Some NLPs induce a hypersensitive‐like response in plants, although the basis for initiation of this response remains unclear. Similarly, the cellular role of such highly conserved proteins is undetermined. It is not clear whether the NLPs are dedicated elicitors of plant defences or whether this induction occurs as a result of another activity.
Pathophysiological characteristics of patients with non‐erosive reflux disease differ from those of patients with functional heartburn
Authors: Frazzoni M. , De Micheli E. , Zentilin P. , Savarino V.
Published: July 2004

Background : Patients with endoscopy‐negative heartburn can be subdivided into non‐erosive reflux disease and functional heartburn on the basis of abnormal and normal, respectively, oesophageal acid exposure. Different pathophysiological characteristics could explain the reportedly low efficacy of proton pump inhibitors in functional heartburn.

Aim : To assess if non‐erosive reflux disease and functional heartburn are pathophysiologically distinguishable.

Methods : Oesophageal manometry and pH‐monitoring were performed in 145 patients with endoscopy‐negative heartburn, in 72 patients with erosive reflux disease, in 58 patients with complicated reflux disease, and in 60 controls.

Results : Patients with non‐erosive reflux disease (84 cases) and functional heartburn (61 cases) differed with regard to the prevalence of hiatal hernia (49% vs. 31%, P = 0.008), the mean lower oesophageal sphincter tone (18.5 vs. 28.4 mmHg, P < 0.05), and the number of upright diurnal acid refluxes lasting more than 5 min (3.6 vs. 0.37, P < 0.05). The results were very close in thenon‐erosive reflux disease, erosive reflux disease and complicated reflux disease groups, whilst patients with functional heartburn were indistinguishable from controls.

Conclusions : Pathophysiological characteristics typical of gastro‐oesophageal reflux disease are found in patients with non‐erosive reflux disease but not in patients with functional heartburn. This could explain the reportedly low efficacy of proton pump inhibitors in functional heartburn and suggests considering different management strategies.
The Visceral Sensitivity Index: development and validation of a gastrointestinal symptom‐specific anxiety scale
Authors: Labus J. S. , Bolus R. , Chang L. , Wiklund I. , Naesdal J. , Mayer E. A. , Naliboff B. D.
Published: July 2004

Background : Anxiety related to gastrointestinal sensations, symptoms or the contexts in which these may occur is thought to play a significant role in the pathophysiology as well as in the health outcomes of patients with irritable bowel syndrome.

Aim : To develop a valid and reliable psychometric instrument that measures gastrointestinal symptom‐specific anxiety.

Methods : External and internal expert panels as well as a patient focus group evaluated a large pool of potential item stems gathered from the psychological and gastrointestinal literature. Potential scale items were then administered to 96 patients diagnosed with irritable bowel syndrome along with a set of validating questionnaires. Final item selection was based upon rigorous empirical criteria and the psychometric properties of the final scale were examined.

Results : A final unidimensional 15‐item scale, the Visceral Sensitivity Index, demonstrated excellent reliability as well as good content, convergent, divergent and predictive validity.

Conclusions : The findings suggest that the Visceral Sensitivity Index is a reliable, valid measure of gastrointestinal symptom‐specific anxiety that may be useful for clinical assessment, treatment outcome studies, and mechanistic studies of the role of symptom‐related anxiety in patients with irritable bowel syndrome.
Pegylated interferon‐α2a kinetics during experimental haemodialysis: impact of permeability and pore size of dialysers
Authors: Barril G. , Quiroga J. A. , Sanz P. , Rodrìguez‐Salvanés F. , Selgas R. , Carreño V.
Published: July 2004

Background : Therapeutics in end‐stage renal disease (ESRD) patients undergoing haemodialysis (HD) has to consider potential drug clearance during the dialysis procedure. Pegylated interferon‐α (PEG‐IFN‐α), a middle‐size protein drug active against viral hepatitis, allows convenient once‐weekly dosing due to prolonged plasma half‐life.

Aim : To investigate the impact of permeability and dialyser pore size on PEG‐IFN‐α blood levels during experimental HD.

Methods : Polymethylmetacrylate (PMMA) membrane 1.6 m2 dialysers with three different permeabilities/pore sizes were selected.

Results : A 40 kDa PEG‐IFN‐α2a (PEGASYS) was not cleared (< 5%) through low‐flux/small pore size (25 Å;B3A) and high‐flux/middle‐large pore size (60 Å;BKP) dialysers, and was partially (≈15%) through intermediate permeability/large pore size (100 Å;BKF) dialysers. In contrast, unmodified 17 kDa IFN‐α2a(Roferon‐A) was removed (65%–95%) through BKP or BKF, but not B3A, PMMA dialysers. Moreover, 12 kDaPEG‐IFN‐α2b(PegIntron) was cleared (40%–80%) through PMMA dialysers with pore sizes ≥ 60 Å. When B3A or BKP were replaced every hour PEG‐IFN‐α2a plasma levels remained constant throughout three experimental‐HD‐sessions, but PEG‐IFN‐α2b was cleared partially every BKP replacement. Porosity differ among high‐flux dialysers. Neither PEG‐IFN‐α2a nor PEG‐IFN‐α2b were removed after three HD sessions through (27/31/33 Å) pore size polysulphone dialysers. Although PEG‐IFN‐α2a was not cleared through middle pore‐size (43 Å/AN69ST) polyacrylonitrile dialyser, PEG‐IFN‐α2b was partially removed.

Conclusions : The pharmacokinetics of Peg‐IFN‐α may vary in a patient on dialysis.
Targeted screening for hereditary haemochromatosis in high‐risk groups
Authors: DuBois S. , Kowdley K. V.
Published: July 2004

Patients with hereditary haemochromatosis are at risk for significant morbidity from iron overload as well as reduced life‐expectancy once cirrhosis is established. Although inexpensive, sensitive screening tests and effective therapy are available, there is continued debate regarding the utility of screening for this condition because of recent data suggesting that the homozygous haemochromatosis mutation (C282Y) is associated with low penetrance and mild expressivity when identified in population screening studies. In this review, we examine the published data related to general population screening for haemochromatosis, as well as the evidence for screening selected ‘high‐risk’ populations. We also suggest possible screening strategies based on the available evidence.
Seven‐day therapy for Helicobacter pylori in the United States
Authors: Vakil N. , Lanza F. , Schwartz H. , Barth J.
Published: July 2004

Background : The ideal duration of Helicobacter pylori treatment in the United States and whether eradication therapy is as successful in nonulcer dyspepsia as in peptic ulcer disease are controversial topics.

Aim : This study compared the efficacy of 3‐, 7‐ and 10‐day triple therapies with rabeprazole to a 10‐day omeprazole control triple therapy for the eradication of Helicobacter pylori in patients with and without peptic ulcer disease in the United States.

Methods : This was a multicentre, double‐blind, randomized, parallel‐group trial. A total of 803 patients with H. pylori infection (determined by [13C]urea breath test and rapid urease test or culture) received either rabeprazole 20 mg b.d., amoxicillin 1000 mg b.d., and clarithromycin 500 mg b.d. for 3, 7, or 10 days, or 10 days of omeprazole 20 mg b.d. with the same antibiotic regimen (control). H. pylori status was assessed by [13C]urea breath test ≥6 weeks after completing treatment.

Results : In intent‐to‐treat patients, the eradication percentages achieved for the rabeprazole‐based treatments were: 3‐day, 27% (95% confidence interval: 21%–34%); 7‐day, 77% (95% confidence interval: 71%–83%); and 10‐day, 78% (95% confidence interval: 72%–84%). The eradication percentage with the 10‐day omeprazole‐based treatment was 73% (95% confidence interval: 67%–79%). There was no statistically significant difference between the 7‐day rabeprazole‐based regimen and the 10‐day rabeprazole‐ and omeprazole‐based regimens.

Conclusions : Seven‐day therapy with rabeprazole, clarithromycin, and amoxicillin is similar in efficacy to 10‐day therapies and had similar efficacy in patients with and without ulcer disease.
Myeloma cells can directly contribute to the pool of RANKL in bone bypassing the classic stromal and osteoblast pathway of osteoclast stimulation
Authors: Lai F. P. L. , Cole‐Sinclair M. , Cheng W.‐J. , Quinn J. M. W. , Gillespie M. T. , Sentry J. W. , Schneider H.‐G.
Published: July 2004

The ratio of osteoprotegerin [OPG, tumour necrosis factor receptor superfamily, member 11b (TNFRSF11B)] to receptor activator of nuclear factor κB ligand [RANKL, tumour necrosis factor (ligand) superfamily, member 11 (TNFSF11)] in bone is critical for the regulation of bone remodelling. Myeloma cells can home to bone, triggering increased RANKL and decreased OPG expression by stromal cells, leading to osteolysis. Whether myeloma cells contribute directly to the pool of RANKL or OPG in bone has been contentious. Here we provide evidence of RANKL expression by reverse transcription polymerase chain reaction and in situ hybridization, demonstrating transcripts encoding both the membrane‐bound and secreted forms of RANKL in five human multiple myeloma cell lines (LP‐1, NCI‐H929, OPM‐2, RPMI8226, U266) and myeloma cells purified from bone marrow aspirates of myeloma patients. We demonstrated that RANKL encoding mRNAs are translated to protein by antibody detection of RANKL. In vitro assays showed that myeloma cells induced bone marrow derived mononuclear cells to differentiate into adherent tartrate‐resistant acid phosphatase positive multinucleated cells, indicative of the formation of functional osteoclasts. This differentiation could also be achieved with passaged myeloma media alone, implicating secreted products. Finally, we provide evidence that the differentiation observed is at least in part the result of myeloma cell expression of RANKL. We therefore conclude that myeloma cells can directly contribute to the pool of RANKL in bone.
Molecular basis of Japanese variants of pyrimidine 5′‐nucleotidase deficiency
Authors: Kanno Hitoshi , Takizawa Takenori , Miwa Shiro , Fujii Hisaichi
Published: July 2004

The type‐I isoform of pyrimidine 5′‐nucleotidase (P5N‐I) has an important role in the catabolism of pyrimidine mononucleotides during erythroid maturation. Two alternatively spliced forms of P5N‐I mRNA have been identified, and we found another alternatively spliced form in reticulocytes, which included an additional 87‐bp sequence. The sequence is located 6·2‐kb downstream of the exon 2 and 2·7‐kb upstream of the exon 3 sequence; consequently, the P5N‐I gene encodes 11 exons, which span approximately 48 kb. We identified five novel mutations in nine families with P5N‐I deficiency: two missense mutations (425C, 721C), one splice mutation (339C), one 1‐bp insertion (251‐insA‐252) and one 9‐bp deletion (del 192‐200). All patients were homozygous for each mutation. The mutant P5N‐I with 721C (G241R) had lower affinity for cytidine monophosphate, suggesting that Gly241 is important for substrate binding. Haplotype analysis showed that 721C, which had been identified in five unrelated families, was a founder mutation. The mutant P5N was then expressed in Cos‐7. The degradation of P5N with 425C (L142P) was significantly faster than a wild‐type control, and proteasome inhibitors restored the stability of L142P. These data suggest that L142P increases susceptibility to the degradation by the ubiquitin‐proteasome pathway.
In search of perfect motherhood for imperfect childhood – experiences of 22 Chinese mothers
Authors: Pun S. H. , Ma Joyce L. C. , Lai Kelly C. C.
Published: July 2004

Motherhood is subjectively defined and contextually based. This article describes and unfolds the experience of 22 Chinese mothers in Hong Kong in cultivating a sense of perfect motherhood for their children who have mental or behavioural problems that require psychiatric consultation. The mother, the significant others and the general public perceive the latter as a form of imperfection. The mothering experiences were full of frustrations, ambivalence, stresses and tensions, and were crystallized by the repeated theme of mother‐blaming and self‐blame by the mother. Despite the hardships, these Chinese mothers have gradually developed self‐confidence and pride, and have found ways to cope with the burden of care. Implications for practice are highlighted at the end of the paper.
Effectiveness of factor VIII infusions in haemophilia A patients with high responding inhibitors
Authors: Kasuda S. , Tanaka I. , Shima M. , Matsumoto T. , Sakurai Y. , Nishiya K. , Giles A. R. , Yoshioka A.
Published: July 2004

Summary. We report here the efficacy of factor VIII (FVIII) infusions in two haemophiliacs with inhibitors using clot waveform analysis on the MDA®II system, which was possible to detect very low levels of FVIII activity < 1.0 U dL−1. In the presence of type 1 inhibitors at the level of 6.2 (patient 1) and 14.4 (patient 2) Bethesda Units mL−1, 3.2 and 6.5 U dL−1of FVIII:C remained 30 min after the infusion of FVIII (100 U kg−1), respectively. Moreover, 0.9 U dL−1of FVIII:C remained 24 h after infusion in patient 2. In both cases, these changes were reflected by qualitative improvement in the aPTT clot waveform and quantitative changes in the minimum value of the second derivative of the aPTT waveform (Min2) that reflects clot acceleration. These results suggest that FVIII infusion may be continued with clinical benefit in some haemophiliacs with high responding inhibitors. Furthermore, the haemostatic response may be monitored accurately and efficiently by clot waveform analysis.
Plasma for fractionation: safety and quality issues
Authors: Farrugia A.
Published: July 2004

Summary.  Plasma may be procured for use as a therapeutic product or as a raw material for manufacture of other products, and may be collected as a by‐product of whole blood, or as a plasma donation from aphaeresis. When collected for fractionation, the quality and safety of the plasma are intimately linked to the quality and safety of the manufactured plasma derivatives. High quality plasma can be obtained either from whole blood or from plasmapheresis; quality can, however, be adversely affected by poor storage conditions after collection. Quality standards for plasma for fractionation are necessarily different than for plasma for transfusion and, with modern fractionation methods, certain quality aspects become less relevant. Similarly, the relevance of certain recent technological advances, such as nucleic acid testing (NAT), for maximizing the safety of plasma for fractionation are questionable, although their introduction through the linkage of recovered plasma to whole blood collection can improve the safety of fresh blood components. Viruses that are not screened for at blood banks may also be excluded from the plasma pool they are more clinically relevant when multiple products made from a pool may infect a large number of recipients, in contrast to components given to one or a small number of patients.
Sequential therapy with activated prothrombin complex concentrate and recombinant factor VIIa in patients with severe haemophilia and inhibitors
Authors: Schneiderman J. , Nugent D. J. , Young G.
Published: July 2004

Summary.  Patients with haemophilia and inhibitors have bleeding episodes that can be refractory to home therapy with either activated prothrombin complex concentrate (APCC) or recombinant factor VIIa (rFVIIa). Sequential therapy with these products has not been widely used because of concern regarding the possibility of thrombosis. This report describes the results of a retrospective chart review of five hospitalized children with severe haemophilia and inhibitors who have been treated with sequential doses of APCC and rFVIIa for refractory bleeding. These patients all had failed home therapy with APCC and rFVIIa alone. A total of 20 admissions were documented covering 170 hospital days, including 91 days of combination therapy. While being closely monitored in the hospital, they received alternating doses of APCC and rFVIIa every 6 h. Anywhere from one to three doses of rFVIIa were given every 2 h between APCC doses. Doses of APCC ranged from 35 to 80 U kg−1 dose−1, and doses of rFVIIa ranged from 80 to 225 mcg kg−1 dose−1. There was no clinical or laboratory evidence of thrombosis, thrombocytopenia, or disseminated intravascular coagulation (DIC). We found the combination of these factors to be safe and effective for patients with refractory bleeds. However, we recommend this aggressive therapy only in the inpatient setting with careful monitoring of the physical examination and frequent laboratory screening to assess for thrombosis and DIC, and without the concurrent use of antifibrinolytic medications.
Successful induction of immune tolerance with FIX recombinant in a patient with haemophilia B with inhibitor
Authors: Soto I. , Vicente J. M. , Corte J. R. , Urgellés M. F. , Pinto C. R.
Published: July 2004

Summary.  We describe successful induction of immune tolerance in a 12‐month‐old patient with severe haemophilia B and low‐titre and low response antifactor IX inhibitor at 5 Bethesda Unit (BU). Immune tolerance induction (ITI) was started with recombinant factor (rFIX) at 40–50 U kg−1 day−1 (1000 U three times per week) and 2 years after the beginning of therapy the inhibitor had been reduced to undetectable values. As complications, which appeared during the ITI, the patient presented minor haemorrhagic complications, which remitted following the administration of rFVIIa, and recurring infections of the catheter, requiring the withdrawal and subsequent placement of a new catheter.
Increased width of the apical periodontal membrane space in endodontically treated teeth may represent favourable healing
Authors: Halse A. , Molven O.
Published: July 2004

Aim  To examine cases showing increased width of the periodontal space (IW) at a long‐term follow‐up examination and to determine whether this finding could be explained by endodontic or nonendodontic factors.

Methodology  A series of intraoral radiographs was obtained from 131 patients 20–27 years after root canal treatment. The same individuals had been examined 10 years earlier, and radiographs taken immediately after treatment were also available. Fourteen roots (5.6%) demonstrated increased width of the apical periodontal space at the end of the study period. These were subjected to further analysis in an attempt to disclose possible explanatory factors.

Results  Two of the 14 cases had reduced marginal bone levels interpreted as the origin of the IW. In three cases, overextended root filling material present 10 years earlier had disappeared and the persistent IW was interpreted as representing a remodelling process. In six cases the findings were explained as being caused by physical and anatomical factors that represented healing without complete re‐formation of the apical periodontal structures, or both. Three cases were judged as unfavourable, on the basis of lacking progress in healing, unsatisfactory obturation of the apical portion of the root canal or dentine resorption close to the apical end of the root filling.

Conclusion  Most of the 14 IW cases examined after 20–27 years could be explained by reduced marginal bone support, or by physical and anatomical factors or they might represent incomplete reformation of the typical apical morphology and were thereby recorded as favourable outcomes.
Delayed‐relaxed response explained by hyperactivation of RelE
Authors: Christensen Susanne K. , Gerdes Kenn
Published: July 2004

Escherichia coli encodes two rel loci, both of which contribute to the control of synthesis of macromolecules during amino acid starvation. The product of relA (ppGpp synthetase I) is responsible for the synthesis of guanosine tetraphosphate, ppGpp, the signal molecule that exerts stringent control of stable RNA synthesis. The second rel locus, relBE, was identified by mutations in relB that confer a so‐called ‘delayed‐relaxed response’ characterized by continued RNA synthesis after a lag period of ≈ 10 min after the onset of amino acid starvation. We show here that the delayed‐relaxed response is a consequence of hyperactivation of RelE. As in wild‐type cells, [ppGpp] increased sharply in relB101 relE cells after the onset of starvation, but returned rapidly to the prestarvation level. RelE is a global inhibitor of translation that is neutralized by RelB by direct protein–protein interaction. Lon protease activates RelE during amino acid starvation by degradation of RelB. We found that mutations in relB that conferred the delayed‐relaxed phenotype destabilized RelB. Such mutations confer severe RelE‐dependent inhibition of translation during amino acid starvation, indicating hyperactivation of RelE. Hyperactivation of RelE during amino acid starvation was shown directly by measurement of RelE‐mediated cleavage of tmRNA. The RelE‐mediated shutdown of translation terminated amino acid consumption and explains the rapid restoration of the ppGpp level observed in relB mutant cells. Restoration of the prestarvation level of ppGpp, in turn, allows for the resumption of stable RNA synthesis seen during the delayed‐relaxed response.
Intracellular effectors regulating the activity of the Lactococcus lactis CodY pleiotropic transcription regulator
Authors: Petranovic Dina , Guédon Eric , Sperandio Brice , Delorme Christine , Ehrlich Dusko , Renault Pierre
Published: July 2004

CodY is a pleiotropic transcriptional regulator conserved in low‐G+C Gram‐positive bacteria. Two distinct signals have been shown independently to influence the activity of this regulator: the level of intracellular GTP in Bacillus subtilis and the level of intracellular branched‐chain amino acids (BCAA) isoleucine, leucine and valine in Lactococcus lactis. Measurement of BCAA and GTP levels in several environmental conditions showed that L. lactis CodY responded to the intracellular BCAA concentrations but not to physiological fluctuations in intracellular GTP. In addition, we demonstrated that CodY from L. lactis did not respond to intracellular GTP even when complementing CodY activity in B. subtilis. However, L. lactis CodY activity could still be modulated in B. subtilis by adding a rich nitrogen source to the growth media. This finding suggests that only BCAA are sensed by L. lactis CodY, whereas both GTP and BCAA signals may be integrated by B. subtilis CodY. The difference in the function of CodY from B. subtilis and L. lactis seems to reflect the difference in the physiology of these two bacteria.
Identification of a 67‐amino‐acid region of the Plasmodium falciparum variant surface antigen that binds chondroitin sulphate A and elicits antibodies reactive with the surface of placental isolates
Authors: Gamain Benoit , Smith Joseph D. , Avril Marion , Baruch Dror I. , Scherf Artur , Gysin Jurg , Miller Louis H.
Published: July 2004

The complications of malaria in pregnancy are caused by the massive sequestration of parasitized erythrocytes (PE) in the placenta. Placental isolates of Plasmodium falciparum are unusual in that they do not bind the primary microvasculature receptor CD36 but instead bind chondroitin sulphate A (CSA). Pregnant mothers develop antibodies that recognize placental variants worldwide, suggesting that a vaccine against malaria in pregnancy is possible. Some members of the Duffy binding‐like γ (DBL‐γ) domain of the large and diverse P. falciparum erythrocyte membrane protein‐1 (PfEMP‐1) family, when expressed on Chinese hamster ovary (CHO) cells, bind CSA. To characterize better the molecular requirements for DBL‐γ adhesion to CSA, we determined the binding of various DBL‐γ domains. Most DBL‐γ did not bind CSA, and no conserved region was identified that strictly differentiated binders from non‐binders. Structure–function analysis of the FCR3‐CSA DBL‐γ domain localized the minimal CSA binding region to a 67‐residue fragment. This region was partially conserved among some binding sequences. Serum from a rabbit immunized with the minimal domain reacted with CSA‐binding parasite lines, but not with non‐CSA‐adherent PE lines that adhered to CD36 and other receptors. The identification of a minimal binding region from a highly variable cytoadherent family may have application for a vaccine against malaria in pregnancy.
Interactions underlying assembly of the Escherichia coli AcrAB–TolC multidrug efflux system
Authors: Touzé Thierry , Eswaran Jeyanthy , Bokma Evert , Koronakis Eva , Hughes Colin , Koronakis Vassilis
Published: July 2004

The major Escherichia coli multidrug efflux pump AcrAB–TolC expels a wide range of antibacterial agents. Using in vivo cross‐linking, we show for the first time that the antiporter AcrB and the adaptor AcrA, which form a translocase in the inner membrane, interact with the outer membrane TolC exit duct to form a contiguous proteinaceous complex spanning the bacterial cell envelope. Assembly of the pump appeared to be constitutive, occurring in the presence and absence of drug efflux substrate. This contrasts with substrate‐induced assembly of the closely related TolC‐dependent protein export machinery, possibly reflecting different assembly dynamics and degrees of substrate responsiveness in the two systems. TolC could be cross‐linked independently to AcrB, showing that their large periplasmic domains are in close proximity. However, isothermal titration calorimetry detected no interaction between the purified AcrB and TolC proteins, suggesting that the adaptor protein is required for their stable association in vivo. Confirming this view, AcrA could be cross‐linked independently to AcrB and TolC in vivo, and calorimetry demonstrated energetically favourable interactions of AcrA with both AcrB and TolC proteins. AcrB was bound by a polypeptide spanning the C‐terminal half of AcrA, but binding to TolC required interaction of N‐ and C‐terminal polypeptides spanning the lipoyl‐like domains predicted to present the intervening coiled‐coil to the periplasmic coils of TolC. These in vivo and in vitro analyses establish the central role of the AcrA adaptor in drug‐independent assembly of the tripartite drug efflux pump, specifically in coupling the inner membrane transporter and the outer membrane exit duct.
Channels in microbes: so many holes to fill
Authors: Kung Ching , Blount Paul
Published: July 2004

Among players in neurobiology, ion channels are the demigods that underlie all our senses, behaviour and intelligence. In animals, these ‘gated pores’ detect ligands, voltage, heat or stretch forces and emit electric or ionic signals. Patch clamp and genome sequencing now show that nearly all microbes also have these ‘smart’ molecules. Microbial channel proteins have yielded crystal structures so dear to neuroscientists. However, their natural roles in microbial physiology remain largely unknown. The intellectual and technical schisms between ‘neuro’ and ‘micro’ biology must be bridged before we know how we became so smart, and whether microbes are just as smart.
Low temperature effects on photosynthesis and growth of grapevine
Authors: HENDRICKSON L. , BALL M. C. , WOOD J. T. , CHOW W. S. , FURBANK R. T.
Published: July 2004

Growth and photosynthesis of grapevine (Vitis vinifera L.) planted on two sloping cool climate vineyards were measured during the early growth season. At both vineyards, a small difference in mean minimum air temperature (1–3 °C) between two microsites accumulated over time, producing differences in shoot growth rate. The growth rates of the warmer (upper) microsite were 34–63% higher than the cooler (lower) site. Photosynthesis measurements of both east and west canopy sides revealed that the difference in carbon gain between the warmer and cooler microsites was due to low temperatures restricting the photosynthetic contribution of east‐facing leaves. East‐facing leaves at the warmer microsite experienced less time at suboptimal temperature while being exposed to high irradiance, contributing to an average 10% greater net carbon gain compared to the east‐facing leaves at the cooler microsite. This chilling‐induced reduction in photosynthesis was not due to net photo‐inhibition. Further analysis revealed that CO2‐ and light‐saturated photosynthesis of grapevines was restricted by stomatal closure from 15 to 25 °C and by a limitation of RuBP regeneration and/or end‐product limitation from 5 to 15 °C. Changes in photosynthetic carboxylation efficiency implied that Rubisco activity may also play a regulatory role at all temperatures. This restriction of total photosynthetic carbon gain is proposed to be a major contributor to the temperature dependence of growth rate at both vineyards during the early season growth period.
The GHOST terminal oxidase regulates developmental programming in tomato fruit
Authors: BARR J. , WHITE W. S. , CHEN L. , BAE H. , RODERMEL S.
Published: July 2004

The tomato ghost and Arabidopsis immutans variegation mutants define orthologous genes for a chloroplast quinol oxidase that bears similarity to the mitochondrial alternative oxidase. In this study tomato fruit ripening was used as a model to explore the function of this oxidase. It was found that a lack of GH impacts the biogenesis of chloroplasts and chromoplasts, as well as pericarp tissue morphogenesis, during the ripening process. Most ripening‐related genes are expressed normally during gh fruit development, but late in the process there is a specific repression of mRNA accumulation from carotenoid biosynthetic genes, perhaps due to retrograde (plastid‐to‐nucleus) signalling pathways mediated by lycopene. Metabolic profiling revealed that wild‐type red‐ripe fruit accumulate multiple isomers of phytoene, phytofluene, and ζ‐carotene, but that the composition of these isomers is different in ripe gh fruit. Profiling  also  revealed  that  there  is  an  enhanced  ratio  of β‐carotene to lycopene in the ripe gh fruit. This explains the orange colour of these fruit, and is consistent with the proposal that the β‐cyclization step plays a pivotal role in regulating the partitioning of pathway intermediates. The total carotenoid content of the mutant fruit is significantly higher than normal due to an accumulation of phytoene. Phytoene accumulation, together with an up‐regulation of GH transcripts during ripening, supports the notion that GH plays a central role in carotenogenesis in chromoplasts. However, GH mRNAs are also expressed in tissues and organs other than fruit. This is consistent with the possibility that GH mediates quinol oxidation from a variety of pathways that converge on plastoquinol in plastid membranes.
Short‐term effects of fertilization on loblolly pine (Pinus taedaL.) physiology
Authors: GOUGH C. M. , SEILER J. R. , MAIER C. A.
Published: July 2004

Fertilization commonly increases biomass production in loblolly pine (Pinus taedaL.). However, the sequence of short‐term physiological adjustments allowing for the establishment of leaf area and enhanced growth is not well understood. The effects of fertilization on photosynthetic parameters, root respiration, and growth for over 200 d following the application of diammonium phosphate were intensively investigated in an effort to establish a relative sequence of events associated with improved growth. Root respiration, foliar nitrogen concentration [N]f, and light‐saturated net photosynthesis (Asat) temporarily increased following fertilization.Asatwas correlated positively with [N]fwhen non‐fertilized and fertilized treatments were pooled (R2 = 0.47). Increased photosynthetic capacity following fertilization was due to both improved photochemical efficiency and capacity and enhanced carboxylation capacity of Rubisco. Positive effects of fertilization on growth were observed shortly afterAsatincreased. Fertilized seedlings had 36.5% more leaf area and 36.5% greater total dry weight biomass at 211 d following fertilization. It is concluded that fertilization temporarily increased photosynthetic capacity, which resulted in a pool of photo‐assimilate used to build leaf area. The N from fertilizer initially invested in photosynthetic structures and enzymes probably re‐translocated to newly developing foliage, explaining the reduction in [N]fandAsatthat was observed after peak levels were achieved following fertilization.