bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinically relevant drug interactions with antiepileptic drugs
Authors: Perucca Emilio
Published: October 2005

Some patients with difficult‐to‐treat epilepsy benefit from combination therapy with two or more antiepileptic drugs (AEDs). Additionally, virtually all epilepsy patients will receive, at some time in their lives, other medications for the management of associated conditions. In these situations, clinically important drug interactions may occur. Carbamazepine, phenytoin, phenobarbital and primidone induce many cytochrome P450 (CYP) and glucuronyl transferase (GT) enzymes, and can reduce drastically the serum concentration of associated drugs which are substrates of the same enzymes. Examples of agents whose serum levels are decreased markedly by enzyme‐inducing AEDs, include lamotrigine, tiagabine, several steroidal drugs, cyclosporin A, oral anticoagulants and many cardiovascular, antineoplastic and psychotropic drugs. Valproic acid is not enzyme inducer, but it may cause clinically relevant drug interactions by inhibiting the metabolism of selected substrates, most notably phenobarbital and lamotrigine. Compared with older generation agents, most of the recently developed AEDs are less likely to induce or inhibit the activity of CYP or GT enzymes. However, they may be a target for metabolically mediated drug interactions, and oxcarbazepine, lamotrigine, felbamate and, at high dosages, topiramate may stimulate the metabolism of oral contraceptive steroids. Levetiracetam, gabapentin and pregabalin have not been reported to cause or be a target for clinically relevant pharmacokinetic drug interactions. Pharmacodynamic interactions involving AEDs have not been well characterized, but their understanding is important for a more rational approach to combination therapy. In particular, neurotoxic effects appear to be more likely with coprescription of AEDs sharing the same primary mechanism of action.
The BRAFV600E mutation is not associated with poor prognostic factors in Korean patients with conventional papillary thyroid microcarcinoma
Authors: Kim Tae Yong , Kim Won Bae , Song Ja Young , Rhee Yoon Soo , Gong Gyungyub , Cho Yong Mee , Kim Sang Yoon , Kim Seong Chul , Hong Suck Joon , Shong Young Kee
Published: October 2005

Background  The BRAFV600E mutation, the most common genetic alteration reported in papillary thyroid carcinoma, has been associated with poor prognostic factors.

Aim  To determine whether the presence of the BRAFV600E mutation is associated with poor prognosis in Korean patients with conventional papillary thyroid microcarcinoma (micro‐PTC).

Patients and methods  DNA was extracted from paraffin‐embedded thyroid tumour specimens taken from 60 patients with conventional micro‐PTC, as well as from nine patients with follicular variant papillary carcinoma, six with nodular hyperplasia, four with follicular carcinoma (including one with Hürthle cell carcinoma), four with follicular adenoma (including two with Hürthle cell adenoma) and one each with medullary carcinoma, poorly differentiated carcinoma and anaplastic carcinoma. The presence of the BRAFV600E mutation was determined by polymerase chain reaction (PCR) amplification of exon 15 followed by direct sequencing.

Results  The BRAFV600E mutation was detected in tumour samples from 31 of 60 conventional micro‐PTC patients (52%), but was not detected in patients with other types of thyroid tumours. The age distribution, tumour size, extrathyroid extension, multifocality and staging did not differ significantly between patients with and without the BRAFV600E mutation.

Conclusion  In Korean patients with conventional micro‐PTC, the presence of the BRAFV600E mutation was not significantly associated with prognostic factors.
Hyporesponsiveness to erythropoietic therapy due to chronic inflammation
Authors: Macdougall I. C. , Cooper A. C.
Published: October 2005

The anaemia associated with chronic renal failure is multi‐factorial. Although a relative erythropoietin deficiency is a major factor, it has also been recognized in recent times that uraemia is a chronic inflammatory state, and thus patients with renal failure also develop anaemia due to mechanisms associated with chronic inflammation. Thus, patients with chronic renal failure have activation of various immune cells, both monocytes and T‐cells. These mononuclear cells have also been shown to release pro‐inflammatory cytokines such as IL‐1, IL‐6, TNF‐alfa and interferon gamma. These cytokines, particularly TNF‐alfa and interferon gamma, are known to cause significant suppression of erythropoiesis. The exact molecular mechanism for this effect is not yet clear, but interferon gamma is an important stimulator of apoptosis in various cell types, including erythroid progenitor cells. This effect may be potentiated by other cytokines such as TNF‐alfa, and this might then antagonise the anti‐apoptotic action of erythropoietin on erythroid progenitors cells, thus reducing responsiveness to exogenous erythropoietic therapy. Chronic renal failure is also associated with increased hepcidin production which may also exacerbate the anaemia by inducing a functional iron deficiency in such patients.
Benefits and risks of iron therapy for chronic anaemias
Authors: Weiss G. , Gordeuk V. R.
Published: October 2005

Iron is used widely for the treatment of anaemias with iron‐restricted erythropoiesis. This intervention can be both beneficial and detrimental depending on the type of the underlying process. While in iron deficiency anaemia (IDA), the most frequent anaemia in the world, iron is the therapy of choice, this intervention can be harmful in the anaemia of chronic disease or anaemia associated with renal failure, the most common anaemias in hospitalized adult patients in Western countries. Iron is able to negatively affect cell‐mediated immune effector mechanisms directed against invading microorganisms and tumour cells while at the same time, as an essential nutrient, it can stimulate the proliferation of these unwanted cells. In addition, iron catalyses the formation of toxic radicals leading to tissue damage or the promotion of cardiovascular events. Thus, it is essential to correctly diagnose the precise cause of anaemia and to consider the benefits and hazards of targeted iron therapy.
Arterial structural and functional alterations in uraemia
Authors: Guérin A. P. , Marchais S. J. , Metivier F. , London G. M.
Published: October 2005

Epidemiological and clinical studies have shown that cardiovascular disease in patients with end‐stage renal disease (ESRD) is frequently related to damage of large conduit arteries. Arterial disease is responsible for the high incidence of ischaemic heart disease, peripheral artery diseases, left ventricular hypertrophy and congestive heart failure. The vascular complications in ESRD are ascribed to two different but associated mechanisms, namely atherosclerosis and arteriosclerosis. Whereas the former principally affects the conduit function with ischaemic lesions being the most characteristic consequence, the latter primarily disturbs the dampening function of large arteries. Arteriosclerosis in ESRD patients is characterized by diffuse dilation and wall hypertrophy of large conduit arteries and stiffening of arterial walls. These changes represent a clinical form of an accelerated ageing process. The main clinical characteristics due to arterial stiffening are isolated increase in systolic blood pressure with normal or lower diastolic pressure resulting in an increased pulse pressure. The consequences of these alterations are: (i) an increased left ventricular afterload with development of left ventricular hypertrophy and increased myocardial oxygen demand; and (ii) altered coronary perfusion and subendocardial blood flow distribution. Epidemiological studies have identified arterial remodelling and stiffening as independent predictors of overall and cardiac mortality in ESRD patients.
The National Anaemia Action Council, Inc.: the primary North American resource for anaemia education and research
Authors: Nissenson A. R.
Published: October 2005

The mission of the National Anaemia Action Council, Inc. (NAAC) is to raise the awareness of health care professionals and the public regarding the prevalence, symptoms, consequences and under‐treatment of anaemia. This editorial illustrates the goals and objectives of NAAC, and depicts past, present and future educational and research projects and findings. NAAC resources in anaemia‐related education and research are described in detail.
Diabetic nephropathy and anaemia
Authors: Ritz E. , Haxsen V.
Published: October 2005

Anaemia is a frequent complication of diabetic nephropathy. It has only recently been recognised that in diabetic patients anaemia is seen not only in preterminal renal failure, but also frequently in patients with only minor derangement of renal function. At any level of glomerular filtration rate (GFR) anaemia is more frequent and severe in diabetic compared to nondiabetic patients. A major cause of anaemia is an inappropriate response of erythropoietin to anaemia. Additional factors are iron deficiency and iatrogenic factors, e.g. ACE inhibitor treatment. When serum creatinine is still normal, the erythropoietin concentration is predictive of more rapid loss of glomerular function. When serum creatinine is elevated, the haemoglobin values are predictive of the rate of progression. It is currently under investigation whether reversal of anaemia attenuates the rate of progression. Because most of the late complications of diabetes (retinopathy, neuropathy, heart disease, peripheral arterial disease) involve ischaemic tissue damage, it would be intuitively plausible that treatment with human recombinant erythropoietin should be beneficial, but definite evidence for this hypothesis is currently not available.
Combined cord blood stem cells and gene therapy enhances angiogenesis and improves cardiac performance in mouse after acute myocardial infarction
Authors: Chen H. K. , Hung H. F. , Shyu K. G. , Wang B. W. , Sheu J. R. , Liang Y. J. , Chang C. C. , Kuan P.
Published: October 2005

Background  Gene and stem cell therapies hold promise for the treatment of ischaemic cardiovascular disease. However, combined stem cell and angiogenic growth factor gene therapy for acute ischaemic myocardium has not been previously reported. This study hypothesized that combined stem cell and gene therapy would not only augment new vessels formation but also improve myocardial function in acute ischaemic myocardium.

Methods  Human angiopoietin‐1 (Ang1) cDNA and VEGF165 cDNA were ligated into AAV vector. The purified CD34+ cells were obtained from human umbilical cord blood samples. Cord blood CD34+ cells were transduced with AAV vector encoding either the human Ang1 (AAV‐Ang1) or VEGF165 (AAV‐VEGF) cDNA alone, or both (AAV‐Ang1 plus VEGF). Immediately after ligation of the left anterior descending coronary artery in male SCID mice, culture‐expanded CD34+ cells transduced with AAV‐Ang1, AAV‐VEGF or AAV‐Ang1 plus VEGF were injected intramyocardially at the left anterior free wall.

Results  Western blot showed that Ang1 and VEGF protein expressions were enhanced in the CD34+cells transduced with AAV‐Ang1 and AAV‐VEGF, respectively. Infarct size significantly decreased and capillary density significantly increased after treatment with CD34+/AAV‐Ang1 plus VEGF when compared with treatment by CD34+ only. Combined therapy with CD34+ and AAV‐Ang1, CD34+ and AAV‐VEGF, CD34+ and AAV‐Ang1 plus VEGF, all showed significantly higher cardiac performance in echocardiography than the therapy with CD34+ alone 4 weeks after myocardial infarction.

Conclusions  Combined therapy with human umbilical cord blood CD34+ cells and both Ang1 and VEGF genes reduced infarct size, attenuated the progression of cardiac dysfunction and increased capillary density in acute myocardial infarction in mice.
Recombinant human erythropoietin: effects on frataxin expression in vitro
Authors: Sturm B. , Stupphann D. , Kaun C. , Boesch S. , Schranzhofer M. , Wojta J. , Goldenberg H. , Scheiber‐Mojdehkar B.
Published: October 2005

Background  Friedreich's ataxia (FRDA) is a neurodegenerative disorder caused by decreased expression of the protein frataxin, recently described to be an iron chaperone for the assembly of iron‐sulphur clusters in the mitochondria, causing iron accumulation in mitochondria, oxidative stress and cell damage. Searching for compounds that could possibly influence frataxin expression, we found that the cytokine recombinant human erythropoietin (rhuEPO) significantly increases frataxin expression by a still unknown mechanism.

Materials and methods  Isolated lymphocytes from FRDA patients, isolated human cardiac cells (fibroblasts and myocytes) from patients undergoing heart transplantation and P19 mouse cells (neuronal typ), were incubated with different concentrations of rhuEPO, and immunoblot was carried out for the detection of frataxin.

Results  We show for the first time that the cytokine recombinant human erythropoietin (rhuEPO) can, additionally to its reported neuro‐ and cardioprotective properties, increase frataxin expression in vitro. We show that rhuEPO significantly increases frataxin expression in primary lymphocytes from patients with Friedreich's ataxia. Further we show that rhuEPO can also increase frataxin expression in many other cell types; among them the most affected cell types in FRDA such as neurones and cardiac cells.

Conclusions  Our results provide a scientific basis for further studies examining the effectiveness of this agent for the treatment of FRDA patients.
Four peptide hormones’ specific decrease (up to 97%) of human prostate carcinoma cells
Authors: Vesely B. A. , Alli A. A. , Song S. J. , Gower W. R. , Sanchez‐Ramos J. , Vesely D. L.
Published: October 2005

Background  Mortality from prostate cancer remains a significant problem with current treatment(s), with an expected 30 350 deaths from prostate cancer in 2005. Long‐acting natriuretic peptide, vessel dilator, kaliuretic peptide and atrial natriuretic peptide have significant anticancer effects in breast and pancreatic adenocarcinomas. Whether these effects are specific and whether they have anticancer effects in prostate adenocarcinoma cells has not been determined.

Materials and methods  These peptide hormones were evaluated to determine if they have specific anticancer effects in human prostate adenocarcinomas.

Results  Dose–response curves revealed a significant (P < 0·05) decrease in human prostate cancer number with each tenfold increase in the concentration from 1 µM to 1000 µM (i.e. 1 mM) of these four peptide hormones. There was a 97·4%, 87%, 88% and 89% (P < 0·001 for each) decrease in prostate cancer cells secondary to vessel dilator, long‐acting natriuretic peptide, kaliuretic peptide and atrial natriuretic peptide, respectively, at their 1‐mM concentrations within 24 h, without any proliferation in the 3 days following this decrease. These same hormones decreased DNA synthesis from 68% to 89% (P < 0·001). When utilized with their respective antibodies their ability to decrease prostate adenocarcinoma cells or inhibit their DNA synthesis was completely blocked. Western blots revealed that for the first time natriuretic peptide receptors (NPR) A‐ and C‐ were present in prostate cancer cells.

Conclusions  These results indicate that these peptide hormones’ anticancer effects are specific. Furthermore, they have very potent effects of eliminating up to 97% of prostate cancer cells within 24 h of treatment.
Cognitive and physiological effects of Omega‐3 polyunsaturated fatty acid supplementation in healthy subjects
Authors: Fontani G. , Corradeschi F. , Felici A. , Alfatti F. , Migliorini S. , Lodi L.
Published: October 2005

Background  It has been reported that Omega‐3 fatty acids may play a role in nervous system activity and that they improve cognitive development and reference memory‐related learning, increase neuroplasticity of nerve membranes, contribute to synaptogenesis and are involved in synaptic transmission. The aim of this study was to examine the effects of Omega‐3 supplementation on some cognitive and physiological parameters in healthy subjects.

Materials and methods  Subjects were tested at the beginning of the experiment and after 35 days. In this period they were supplemented with Omega‐3 polyunsaturated fatty acids. A group was supplemented with olive oil (placebo). Tests involving different types of attention were used, i.e. Alert, Go/No‐Go, Choice and Sustained Attention. For each test, the reaction time, the event‐related potentials by electroencephalogram (EEG) and the electromyography (EMG) of the forefinger flexor muscle were recorded. The Profile of Mood States test (POMS) was also administered.

Results  Blood analyses showed that after Omega‐3 supplementation the arachidonic acid/eicosapentaenoic acid ratio (AA/EPA) was strongly reduced. The mood profile was improved after Omega‐3 with increased vigour and reduced anger, anxiety and depression states. This was associated with an effect on reactivity with a reduction of reaction time in the Go/No‐Go and Sustained Attention tests. The latency of EMG activation was concomitantly reduced in the same tests plus Choice. An EEG frequency shift towards the theta and alpha band were recorded in all the tests after Omega‐3.

Conclusions  Omega‐3 supplementation is associated with an improvement of attentional and physiological functions, particularly those involving complex cortical processing. These findings are discussed in terms of the influence of Omega‐3 on the central nervous system.
Use of major‐ and trace‐element correlations to assess metal migration in sandy Luvisols irrigated with wastewater
Authors: Lamy I. , Van Oort F. , Dère C. , Baize D.
Published: October 2005

Soils irrigated with raw wastewater for many years accumulate trace metals, especially in the surface horizon. We do not know, however, to what extent the metals migrate down the profile and whether they accumulate in deeper horizons. We assessed the migration of trace metals in the suburbs of Paris where market gardens had been irrigated with wastewater for more than 100 years. Within a soil series, we compared metals in the polluted soils with those of a reference soil of the same kind. We sampled by horizons three representative Luvisol profiles in the irrigated area and a fourth profile selected as a reference just outside the irrigated area. Large quantities of Zn, Pb, Cd and Cu appeared to have accumulated in the topsoil, at the base of which concentrations diminished markedly. There appeared to be appreciable migration into the subsoil only at the most intensely irrigated site. By relating the total trace metal contents to the total iron contents we could detect anomalous values and identify migration into the subsurface horizons more sensitively. This approach was tested first with the endogenous metal Cr, then with metal pollutants. We showed that Zn, Pb, Cd and Cu migrated downwards in all three irrigated soils.
Comparing and evaluating process‐based ecosystem model predictions of carbon and water fluxes in major European forest biomes
Authors: Morales Pablo , Sykes Martin T. , Prentice I. Colin , Smith Pete , Smith Benjamin , Bugmann Harald , Zierl Bärbel , Friedlingstein Pierre , Viovy Nicolas , Sabaté Santi , Sánchez Anabel , Pla Eduard , Gracia Carlos A. , Sitch Stephen , Arneth Almut , Ogee Jerome
Published: October 2005

Process‐based models can be classified into: (a) terrestrial biogeochemical models (TBMs), which simulate fluxes of carbon, water and nitrogen coupled within terrestrial ecosystems, and (b) dynamic global vegetation models (DGVMs), which further couple these processes interactively with changes in slow ecosystem processes depending on resource competition, establishment, growth and mortality of different vegetation types. In this study, four models – RHESSys, GOTILWA+, LPJ‐GUESS and ORCHIDEE – representing both modelling approaches were compared and evaluated against benchmarks provided by eddy‐covariance measurements of carbon and water fluxes at 15 forest sites within the EUROFLUX project. Overall, model‐measurement agreement varied greatly among sites. Both modelling approaches have somewhat different strengths, but there was no model among those tested that universally performed well on the two variables evaluated. Small biases and errors suggest that ORCHIDEE and GOTILWA+ performed better in simulating carbon fluxes while LPJ‐GUESS and RHESSys did a better job in simulating water fluxes. In general, the models can be considered as useful tools for studies of climate change impacts on carbon and water cycling in forests. However, the various sources of variation among models simulations and between models simulations and observed data described in this study place some constraints on the results and to some extent reduce their reliability. For example, at most sites in the Mediterranean region all models generally performed poorly most likely because of problems in the representation of water stress effects on both carbon uptake by photosynthesis and carbon release by heterotrophic respiration (Rh).

The use of flux data as a means of assessing key processes in models of this type is an important approach to improving model performance. Our results show that the models have value but that further model development is necessary with regard to the representation of the some of the key ecosystem processes.
Demonstration of EGFR gene copy loss in colorectal carcinomas by fluorescence in situ hybridization (FISH): a surrogate marker for sensitivity to specific anti‐EGFR therapy?
Authors: Sauer T , Guren M G , Noren T , Dueland S
Published: October 2005

Aims : To investigate EGFR gene copy number heterogeneity in colorectal carcinomas compared with copy number of chromosome 7 and immunohistochemical expression of the EGFR protein.

Methods and results : Fluorescence in situ hybridization of the EGFR gene and CEP7 was carried out on paraffin‐embedded material from 48 rectal carcinomas combined with immunohistochemical detection of EGFR with a polymer detection kit. EGFR gene copy number had a range of 1.4–7.3 with a mean of 2.5. CEP7 copy number had a range of 1.5–6.1 with a mean of 2.5. The EGFR gene/CEP7 ratio ranged from 0.4 to 1.5 with a mean of 0.96. Most cases had a balanced EGFR gene/CEP7 ratio (37 cases = 77%). Copy gain was found in seven cases (15%) with a ratio of up to 1.5, consistent with gain of one EGFR gene copy in one chromosome. Copy loss was found in four cases (8%). All cases with EGFR gene copy loss were immunohistochemically positive.

Conclusions : Demonstration of EGFR gene copy loss might be a surrogate marker for EGFR mutation/deletion and could be used in a routine setting in pathology departments. Further studies are needed to determine whether this may be used to select patients that might benefit from specific anti‐EGFR therapy.
The MAPK–AP‐1/–Runx2 signalling axes are implicated in chondrosarcoma pathobiology either independently or via up‐regulation of VEGF
Authors: Papachristou D J , Papachristou G J , Papaefthimiou O A , Agnantis N J , Basdra E K , Papavassiliou A G
Published: October 2005

Aims : To investigate whether and how the JNK/ERK–AP‐1/–Runx2 signalling pathways and vascular endothelial growth factor (VEGF) are engaged in the pathogenesis of cartilaginous tumours. Chondrosarcoma is the third most common primary skeletal malignancy. Nevertheless, the molecular events underlying its pathogenesis remain elusive. JNK/ERK MAPKs and their downstream effectors, c‐Jun and c‐Fos (AP‐1), are involved in chondroblastic differention/proliferation. These proteins interact with the Runx2 transcription factor, which is also implicated in chondroblast biology. VEGF, a key angiogenic factor, is up‐regulated in human chondrosarcomas.

Methods and results : Normal cartilage and neoplastic cells from 45 chondrosarcomas and 21 enchondromas were investigated immunohistochemically. We evaluated the cellular levels of JNK2, p‐JNK2 (phosphorylated/activated JNK2), its main substrate, c‐Jun, pc‐Jun (phosphorylated/activated c‐Jun) and c‐Fos. Moreover, the levels of p‐ERK (phosphorylated/activated ERK), Runx2 and VEGF were assessed. Positive immunostaining for all proteins was observed in the majority of the examined chondrosarcomas and in a small fraction of enchondromas. The expression levels of all proteins were positively and significantly correlated with each other. These levels were substantially augmented in high‐grade compared with low‐grade chondrosarcomas and in low‐grade tumours compared with benign enchondromas, implying a potential use as molecular markers for prediction of high‐grade neoplasms.

Conclusions : The JNK/ERK–AP‐1/–Runx2 signal transduction ‘network’ is associated with chondroblastic malignant transformation and chondrosarcoma development, either separately or through coordinated induction of VEGF.
Molecular evidence supporting the neoplastic nature of odontogenic keratocyst: a laser capture microdissection study of 15 cases
Authors: Henley J , Summerlin D‐J , Tomich C , Zhang S , Cheng L
Published: October 2005

Aims : The bland histology of odontogenic keratocyst (OKC) belies its capacity for aggressive behaviour. Genetic alterations of OKC have not been well studied. We examined the frequency and pattern of allelic imbalance on five different chromosome regions from 15 patients with OKC.

Methods and results : Laser‐assisted microdissection was performed on formalin‐fixed paraffin‐embedded tissue. Polymerase chain reaction analysis of extracted DNA targeted five polymorphic DNA markers (D3S1285, D9S161, D11S1316, D13S290, and TP53) representing chromosome regions 3p14, 9p21, 11q23, 13q12.1 and 17p13, respectively. All 15 cases of OKC were informative at a minimum of three of five loci, with 11 informative on all five loci. Twelve of 15 cases (80%) demonstrated loss of heterozygosity (LOH). Seven cases (47%) showed LOH at more than two DNA loci. The frequency of LOH was 5/11 (45%) at D3S1285, 3/15 (20%) at D9S161, 4/14 (29%) at D11S1316, 8/14 (57%) at D13S290 and 3/15 (20%) at TP53.

Conclusions : The majority of OKCs harbour chromosomal abnormalities. This finding supports the supposition that OKCs are neoplastic. Furthermore, OKCs harbour allelic loss at some of the same loci identified in squamous cell carcinoma. This may aid in explaining the rare occurrence of squamous cell carcinoma arising in OKC.
Use of novel monoclonal antibodies to determine the expression and distribution of the hypoxia regulatory factors PHD‐1, PHD‐2, PHD‐3 and FIH in normal and neoplastic human tissues
Authors: Soilleux E J , Turley H , Tian Y M , Pugh C W , Gatter K C , Harris A L
Published: October 2005

Aims : The cellular response to hypoxia includes the hypoxia inducible factor (HIF)‐induced transcription of genes involved in diverse processes such as glycolysis, angiogenesis and the growth of experimental tumours. Regulation of the level of hypoxia inducible factors 1α and 2α (HIF‐1α and HIF‐2α) is a primary determinant of HIF activity. Recent biochemical and candidate gene approach studies have led to the discovery of three HIF‐regulatory prolyl hydroxylases, PHD‐1, ‐2 and ‐3 and an asparaginyl hydroxylase, also known as FIH (factor inhibiting HIF). In this study, we raised and characterized monoclonal antibodies against PHD‐1, PHD‐2, PHD‐3 and FIH.

Methods and results : Immunohistochemistry of normal tissues with these monoclonal antibodies demonstrated a wide distribution in epithelial cells, stromal cells and leucocytes, with cytoplasmic staining predominating over nuclear staining. A preliminary study of tumours showed variable staining in tumour, stromal and inflammatory cells. While all tumour types showed some positive staining with each antibody, the overall pattern suggested a slight decrease in the amount of staining seen with PHD‐1, ‐2 and ‐3 and an increase in FIH staining in neoplasia compared with corresponding normal tissues.

Conclusions : These monoclonal antibodies will allow further larger scale studies to determine the significance of PHD and FIH expression in neoplasia.
Effect of food ingredients and selected lipids on the physical properties of extruded edible films/casings
Authors: Liu Li , Kerry John F. , Kerry Joe P.
Published: October 2005

Films/casings produced using pectin (PN) and gelatin/sodium alginate blends (GSAB) containing 2.5 and 5% added corn oil (CO) or olive oil (OO) and non‐oil containing were manufactured using extrusion technology. Films/casings formed from PN and GSAB had great difference in properties of tensile strength, elongation, Young's modulus and puncture resistance. Quality and stability of films/casings were greatly enhanced on addition of oils. However, an oil addition of 2.5% was identified as optimal while 5% decreased the final films/casings quality. Oil addition increased (P < 0.05) films/casings thicknesses. Oil addition also decreased (P < 0.05) tensile strength, puncture resistance, Young's modulus and tear resistance of films/casings but increased their elongation properties, with the exception of converse effect of 2.5% CO on two films/casings. Olive oil and CO both reduced (P < 0.05) water vapour transfer rate (WVTR) of films/casings. However, no significant difference between films using different concentrations of oil was observed with respect to WVTR.
Muralytic activity of Micrococcus luteus Rpf and its relationship to physiological activity in promoting bacterial growth and resuscitation
Authors: Mukamolova Galina V. , Murzin Alexey G. , Salina Elena G. , Demina Galina R. , Kell Douglas B. , Kaprelyants Arseny S. , Young Michael
Published: October 2005

The culturability of several actinobacteria is controlled by resuscitation‐promoting factors (Rpfs). These are proteins containing a c. 70‐residue domain that adopts a lysozyme‐like fold. The invariant catalytic glutamate residue found in lysozyme and various bacterial lytic transglycosylases is also conserved in the Rpf proteins. Rpf from Micrococcus luteus, the founder member of this protein family, is indeed a muralytic enzyme, as revealed by its activity in zymograms containing M. luteus cell walls and its ability to (i) cause lysis of Escherichia coli when expressed and secreted into the periplasm; (ii) release fluorescent material from fluorescamine‐labelled cell walls of M. luteus; and (iii) hydrolyse the artificial lysozyme substrate, 4‐methylumbelliferyl‐β‐d‐N,N′,N′′‐triacetylchitotrioside. Rpf activity was reduced but not completely abolished when the invariant glutamate residue was altered. Moreover, none of the other acidic residues in the Rpf domain was absolutely required for muralytic activity. Replacement of one or both of the cysteine residues that probably form a disulphide bridge within Rpf impaired but did not completely abolish muralytic activity. The muralytic activities of the Rpf mutants were correlated with their abilities to stimulate bacterial culturability and resuscitation, consistent with the view that the biological activity of Rpf results directly or indirectly from its ability to cleave bonds in bacterial peptidoglycan.
The chaperone binding domain of SopE inhibits transport via flagellar and SPI‐1 TTSS in the absence of InvB
Authors: Ehrbar Kristin , Winnen Brit , Hardt Wolf‐Dietrich
Published: October 2005

Type III secretion systems (TTSS) are used by many Gram‐negative pathogens for transporting effector proteins into eukaryotic host cells. Two modes of type III effector protein transport can be distinguished: transport into the surrounding medium (secretion) and cell‐contact induced injection of effector proteins directly into the host cell cytosol (translocation). Two domains within the N‐terminal regions of effector proteins determine the mode of transport. The amino terminal ∼20 amino acids (N‐terminal secretion signal, NSS) mediate secretion. In contrast, translocation generally requires the NSS, the adjacent ∼100 amino acids (chaperone binding domain, CBD) and binding of the cognate chaperone to this CBD. TTSS are phylogenetically related to flagellar systems. Because both systems are expressed in Salmonella Typhimurium, correct effector protein transport involves at least two decisions: transport via the Salmonella pathogenicity island 1 (SPI‐1) but not the flagellar TTSS (= specificity) and translocation into the host cell instead of secretion into the surrounding media (= transport mode). The mechanisms guiding these decisions are poorly understood. We have studied the S. Typhimurium effector protein SopE, which is specifically transported via the SPI‐1 TTSS. Secretion and translocation strictly require the cognate chaperone InvB. Alanine replacement of amino acids 30–42 (and to some extent 44–54) abolished tight InvB binding, abolished translocation into the host cell and led to secretion of SopE via both, the flagellar and the SPI‐1 TTSS. In clear contrast to wild‐type SopE, secretion of SopEAla30‐42 and SopEAla44‐54 via the SPI‐1 and the flagellar export system did not require InvB. These data reveal a novel function of the CBD: the CBD inhibits secretion of wild‐type SopE via the flagellar and the SPI‐1 TTSS in the absence of the chaperone InvB. Our data provide new insights into mechanisms ensuring specific effector protein transport by TTSS.
Identification of the central quorum sensing regulator of virulence in the enteric phytopathogen, Erwinia carotovora: the VirR repressor
Authors: Burr Tom , Barnard Anne M. L. , Corbett Mark J. , Pemberton Clare L. , Simpson Natalie J. L. , Salmond George P. C.
Published: October 2005

In the Gram‐negative phytopathogen, Erwinia carotovora ssp. atroseptica (Eca) virulence depends on the production of a N‐(3‐oxohexanoyl)‐l‐homoserine lactone (OHHL) quorum sensing (QS) signal. This work identifies the elusive ‘missing link’ between QS and virulence in Erwinia. We have identified and characterized a novel regulator of virulence, VirR, in Eca and show that a virR mutation completely restores virulence factor production to an Eca mutant unable to synthesize OHHL. This effect of the virR mutation translates to a restoration of virulence to wild‐type levels and thus provides evidence that VirR acts to prevent the production of virulence factors at low cell density. We also show that, in Eca, transcription of virulence genes is controlled by OHHL and that this control is effected through the action of VirR. We also demonstrate that the VirR regulatory pathway is present and functional in both blackleg and soft rotting species of Erwinia.
Virulence and drug resistance roles of multidrug efflux systems of Salmonella enterica serovar Typhimurium
Authors: Nishino Kunihiko , Latifi Tammy , Groisman Eduardo A.
Published: October 2005

Drug efflux systems play a major role in resistance to a wide range of noxious compounds in several Gram negative species. Here, we report the drug resistance and virulence phenotypes of Salmonella mutants defective in either resistance‐nodulation‐division (RND)‐type systems and/or in drug efflux systems belonging to the major facilitator (MFS), multidrug and toxic compound extrusion (MATE), and ATP‐binding cassette (ABC) superfamilies. We determined that nine potential drug transporters contribute to drug resistance of Salmonella and found that the Salmonella‐specific MdsABC system conferred resistance to a variety of toxic compounds. The RND‐type MdsAB system could function with either MdsC, which is encoded in the same operon, or TolC as the outer membrane component. Although the Salmonella EmrAB, MdfA and MdtK are 90% identical in their amino acid sequences to their Escherichia coli homologues, the drug specificity of Salmonella transporters was different from that reported for equivalent E. coli transporters. Deletion of the macAB genes attenuated Salmonella virulence and a strain lacking all drug efflux systems was avirulent when mice were inoculated by the oral route. The promoter region of the macAB drug efflux system genes harbours a binding site for the response regulator PhoP, which functions to repress macAB transcription. The PhoP/PhoQ two‐component system is a major regulator of Salmonella virulence, which underscores the connection between drug efflux systems and virulence.
A small RNA inhibits translation of the histone‐like protein Hc1 in Chlamydia trachomatis
Authors: Grieshaber Nicole A. , Grieshaber Scott S. , Fischer Elizabeth R. , Hackstadt Ted
Published: October 2005

The chromatin of chlamydial elementary bodies (EBs) is stabilized by proteins with sequence homology to eukaryotic H1. These histone homologues, termed Hc1 and Hc2, are expressed only during the late stages of the chlamydial life cycle concomitant with the reorganization of reticulate bodies (RBs) into metabolically inactive EBs. Hc1 and Hc2 play a major role in establishment of nucleoid structure as well as in downregulation of gene expression as RBs differentiate back to EBs. The effects of Hc1 on gene expression patterns requires that chlamydiae strictly control Hc1 activity. Hc1 expression and activity are thus regulated transcriptionally as well as post‐transcriptionally. We describe here a small regulatory RNA (sRNA) that acts as an additional checkpoint to negatively regulate Hc1 synthesis. Coexpression of the sRNA with hctA, the gene that encodes Hc1, in Escherichia coli inhibited Hc1 translation but did not affect hctA mRNA transcription or stability. IhtA (inhibitor of hctA translation) was present only in purified RBs while Hc1 was present only in purified EBs. During infection IhtA, but not Hc1, was present in RBs and was downregulated while Hc1 was upregulated upon RB to EB differentiation. Thus, we propose that IhtA is part of a global regulatory circuit that controls differentiation of RBs to EBs during the chlamydial life cycle.
Hypothalamic‐pituitary gonadal axis and immune response imbalance during chronic filarial infections
Authors: Mavoungou Donatien , Poaty‐Mavoungou Virginie , Ongali Brice , Akoume Marie Yvonne , Maka Gontran , Mavoungou Elie
Published: October 2005

Bi‐directional relationships operate between the hypothalamic‐pituitary‐gonadal axis and the immune system. Cytokines, peptide hormones and their shared receptors/ligands are used as a common biological language for communication within and between the immune and neuroendocrine systems. Such communication suggests an immunoregulatory role for the brain and a sensory function for the immune system. We used a radioimmunoassay to measure the concentrations of steroid hormones (cortisol, testosterone, estradiol and progesterone) and pituitary hormones [follicle stimulating hormone (FSH), luteinizing hormone (LH) human chorionic gonadotropin (HCG) and prolactin] in peripheral blood plasma from 78 young Gabonese women with chronic filarial infections. We used an enzyme‐linked immunosorbent assay to determine the concentrations of four proinflammatory cytokines [tumor necrosis factor‐alpha (TNF‐α), gamma interferon (IFN‐γ), interleukin‐1 (IL‐1) and IL‐6] in the same plasma samples. Progesterone was unchanged and all other steroid hormone plasma concentrations were lower in microfilaremic women than in amicrofilaremic women. The concentration of LH was higher in amicrofilaremic women, whereas the prolactin concentration was higher in microfilaremics. The plasma concentrations of TNF‐α, IFN‐γ, IL‐1 and IL‐6 were higher in microfilaremic women. A strong negative correlation was found between the steroid and pituitary hormones and the pro‐inflammatory cytokines. Conversely, a strong positive correlation was found between prolactin and the same cytokines. These data provide first evidence of immune system and hormonal system disturbance during chronic filarial infections and suggest that the observed imbalance should be taken into account in the diagnosis and treatment of filarial infections.
Cytokine and Ig‐production by CG‐containing sequences with phosphorodiester backbone and dumbbell‐shape
Authors: Schmidt M. , Anton K. , Nordhaus C. , Junghans C. , Wittig B. , Worm M.
Published: October 2005

Background:  Immunostimulatory oligodeoxynucleotides (CpG‐ODN) usually contain phosphorothioate (PS) backbones for nucleotide protection, which may result in some nonspecific side‐effects like prolongation of coagulation time.

Objective:  The aim of the present study was to investigate the immunomodulatory potential of DNA molecules without PS backbones. Thus, we designed phosphorodiester (PO) molecules with a dumbbell‐like covalently‐closed structure (dSLIM‐30L1).

Methods:  We analyzed their effects on peripheral blood mononuclear cells (PBMC) from spontaneous high and low immunoglobulin (Ig)E producer (allergic and nonallergic donors) in comparison with linear CpG‐ODN (lin‐30L1) with PS backbones, using enzyme‐linked immunosorbent assay and flow cytometry.

Results:  We observed a decrease of spontaneous IgE levels in PBMC from high IgE producer of approximately 27% with both dSLIM‐30L1 and lin‐30L1. In addition, both molecules enhanced the production of IgA, IgM and IgG1/IgG2, but with a slightly different pattern. Both molecules stimulated the secretion of the TH1‐like cytokines interleukin (IL)‐2, interferon‐γ and IL‐12p40 and the pro‐inflammatory cytokine IL‐6. The immunomodulatory potential of dSLIM‐30L1 and lin‐30L1 was also effective in PBMC from nonallergic donors, as was confirmed for IL‐2, IL‐12p40, IgG1/IgG2 and IgM.

Conclusion:  Our data show an inhibition of IgE production but also enhancement of the inflammatory cytokine response in PBMC from allergic and nonallergic donors by covalently‐closed PO‐based dSLIM‐30L1 with a pattern similar to that of linear PS‐based lin‐30L1, while avoiding PS‐modifications and thus PS‐mediated side‐effects. Whether such molecules are useful for the treatment of allergic diseases will need further clarification by appropriate in vivo studies.
Dietary intake in sensitized children with recurrent wheeze and healthy controls: a nested case–control study
Authors: Murray C. S. , Simpson B. , Kerry G. , Woodcock A. , Custovic A.
Published: October 2005

Background:  The rising prevalence of asthma and allergic disease remains unexplained. Several risk factors have been implicated including diet, in particular poly‐unsaturated fats and antioxidant intake.

Methods:  A nested case–control study comparing the dietary intake of sensitized children with recurrent wheeze (age 3–5 years) and nonsensitized children who had never wheezed was carried out within an unselected population‐based cohort. Cases and controls were matched for age, sex, parental atopy, indoor allergen exposure and pet ownership. Dietary intake was assessed using a validated semi‐quantitative food frequency questionnaire and nutrient analysis program.

Results:  Thirty‐seven case–control pairs (23 male, mean age 4.4 years) participated. Daily total polyunsaturated fat intake was significantly higher in sensitized wheezers (g/day, geometric mean, 95% confidence intervals: 7.1, 6.4–7.9) compared with nonsensitized nonwheezy children (5.6, 5.0–6.3, P = 0.003). Daily omega‐3 and omega‐6 fat intakes were not significantly different between the two groups. No significant differences were found in intake of any antioxidant or antioxidant cofactors between the groups.

Conclusions:  Young sensitized wheezy children had a significantly higher total polyunsaturated fat intake compared with nonsensitized nonwheezy children. However, we were unable to distinguish a significant difference in specific poly‐unsaturated fat intakes. Otherwise the children in both groups had a very similar nutritional intake.
Comparison of X‐ray vs. gamma irradiation of CPDA‐1 red cells
Authors: Janatpour K. , Denning L. , Nelson K. , Betlach B. , MacKenzie M. , Holland P.
Published: October 2005

Background and Objectives  Transfusion‐associated graft‐versus‐host disease (TA‐GVHD) is a serious, potential complication of blood transfusion that is essentially prevented by blood product irradiation. Blood product irradiation is currently performed using gamma irradiation. X‐ray irradiation is an alternative that has certain advantages: an X‐ray irradiator is less expensive and does not have a radioactive source. However, the biochemical effects of X‐ray irradiation on red blood cells (RBCs) have not been well characterized. The primary purpose of our study was to compare the effects of X‐ray irradiation with gamma irradiation on RBC membrane permeability. A secondary purpose was to verify that X‐ray irradiation has the same effect on lymphocytes as gamma irradiation.

Materials and Methods  Ten RBC units were split into two portions; five units of each were then irradiated with either gamma or X‐ray at a dose of 35 Gy or 25 Gy, respectively. Free plasma haemoglobin and extracellular potassium levels were measured in each unit over time. Another unit was divided into three parts for lymphocyte studies; one part was not irradiated as a control and the other two received 25 Gy of irradiation via X‐ray or gamma irradiation, respectively.

Results  Minimally increased free plasma haemoglobin was found only in X‐ray‐irradiated units after 25 Gy compared to gamma‐irradiated units. However, extracellular potassium levels were not significantly different in these groups. Extracellular potassium levels were slightly higher only in gamma irradiated units at 35 Gy. X‐ray and gamma irradiation were equivalent in affecting lymphocyte function.

Conclusions  Small differences in RBC membrane permeability are found between gamma‐irradiated and X‐ray‐irradiated units. However, these differences are not likely to be clinically important.
Nurses’ moral agency in negotiating parental participation in care
Authors: O'Haire Sophie E , Blackford Jeanine C
Published: October 2005

The purpose of this study was to investigate the issues for nurses in facilitating parental participation in the care of the hospitalized child. A qualitative study informed by grounded theory was undertaken. Nine nurses were recruited from an acute, high‐dependency, 23‐bed paediatric cardiac/renal unit in Melbourne, Australia. Data collection involved individual semistructured interviews, hospital policies related to family‐centred care and a focus group interview. Constant comparative analysis was undertaken to develop an understanding of the data collected in the context of the nurses’ experiences and the environment in which they work. Moral agency was identified as the central phenomenon of the study. Causal conditions related to this included the child's best interests, disputes about care and nurses’ expectations. These causal conditions were seen to potentially lead to moral distress for the nurses. The coping mechanisms and strategies that affected moral agency have been identified.
Improving transfer from the intensive care unit: The development, implementation and evaluation of a brochure based on Knowles’ Adult Learning Theory
Authors: Mitchell Marion L , Courtney Mary
Published: October 2005

This paper describes the development, implementation and evaluation of a transfer brochure for family members of patients in an intensive care unit (ICU) to improve patient transfer to a general ward. When family members fail to understand information, they respond in ways that affect patient recovery. The brochure was designed within Knowles’ Adult Learning Theory framework and developed using a multidisciplinary team. A mixed design was used to collect data from families and nurses. Results indicate that the brochure helped nurses to address the individual family's issues during transfer from ICU. Furthermore, 95% of nurses (n = 33) recommended its introduction for all future transfers. Family members (n = 82) who received the brochure as part of their transfer were significantly more satisfied with all aspects of transfer than those who experienced ad hoc transfer methods (n = 80). These results provide strong support for Knowles’ Adult Learning Theory as an educational foundation for adult learning.