bluuebunny/crossref_metadata_2025 huggingface dataset:
An international sepsis survey: a study of doctors' knowledge and perception about sepsis
Authors: Poeze Martijn , Ramsay Graham , Gerlach Herwig , Rubulotta Francesca , Levy Mitchel
Published: October 2004

Background

To be able to diagnose and treat sepsis better it is important not only to improve the knowledge about definitions and pathophysiology, but also to gain more insight into specialists' perception of, and attitude towards, the current diagnosis and treatment of sepsis.

Methods

The study was conducted as a prospective, international survey by structured telephone interview. The subjects were intensive care physicians and other specialist physicians caring for intensive care unit (ICU) patients.

Results

The 1058 physicians who were interviewed (including 529 intensivists) agreed that sepsis is a leading cause of death on the ICU and that the incidence of sepsis is increasing, but that the symptoms of sepsis can easily be misattributed to other conditions. Physicians were concerned that this could lead to under-reporting of sepsis. Two-thirds (67%) were concerned that a common definition is lacking and 83% said it is likely that sepsis is frequently missed. Not more than 17% agreed on any one definition.

Conclusion

There is a general awareness about the inadequacy of the current definitions of sepsis. Physicians caring for patients with sepsis recognise the difficulty of defining and diagnosing sepsis and are aware that they miss the diagnosis frequently.
Prospective evaluation of an internet-linked handheld computer critical care knowledge access system
Authors: Lapinsky Stephen E , Wax Randy , Showalter Randy , Martinez-Motta J Carlos , Hallett David , Mehta Sangeeta , Burry Lisa , Stewart Thomas E
Published: October 2004

Introduction

Critical care physicians may benefit from immediate access to medical reference material. We evaluated the feasibility and potential benefits of a handheld computer based knowledge access system linking a central academic intensive care unit (ICU) to multiple community-based ICUs.

Methods

Four community hospital ICUs with 17 physicians participated in this prospective interventional study. Following training in the use of an internet-linked, updateable handheld computer knowledge access system, the physicians used the handheld devices in their clinical environment for a 12-month intervention period. Feasibility of the system was evaluated by tracking use of the handheld computer and by conducting surveys and focus group discussions. Before and after the intervention period, participants underwent simulated patient care scenarios designed to evaluate the information sources they accessed, as well as the speed and quality of their decision making. Participants generated admission orders during each scenario, which were scored by blinded evaluators.

Results

Ten physicians (59%) used the system regularly, predominantly for nonmedical applications (median 32.8/month, interquartile range [IQR] 28.3–126.8), with medical software accessed less often (median 9/month, IQR 3.7–13.7). Eight out of 13 physicians (62%) who completed the final scenarios chose to use the handheld computer for information access. The median time to access information on the handheld handheld computer was 19 s (IQR 15–40 s). This group exhibited a significant improvement in admission order score as compared with those who used other resources (P = 0.018). Benefits and barriers to use of this technology were identified.

Conclusion

An updateable handheld computer system is feasible as a means of point-of-care access to medical reference material and may improve clinical decision making. However, during the study, acceptance of the system was variable. Improved training and new technology may overcome some of the barriers we identified.
Place of death: hospital-based advanced home care versus conventional care
Authors: Ahlner-Elmqvist Marianne , Jordhøy Marit S , Jannert Magnus , Fayers Peter , Kaasa Stein
Published: October 2004

The purpose of this prospective nonrandomized study was to evaluate time spent at home, place of death and differences in sociodemographic and medical characteristics of patients, with cancer in palliative stage, receiving either hospital-based advanced home care (AHC), including 24-hour service by a multidisciplinary palliative care team or conventional hospital care (CC). Recruitment to the AHC group and to the study was a two-step procedure. The patients were assigned to either hospital-based AHC or CC according to their preferences. Following this, the patients were asked to participate in the study. Patients were eligible for the study if they had malignant disease, were older than 18 years and had a survival expectancy of 2-12 months. A total of 297 patients entered the study and 280 died during the study period of two and a half years, 117 in the AHC group and 163 in the CC group. Significantly more patients died at home in the AHC group (45%) compared with the CC group (10%). Preference for and referral to hospital-based AHC were not related to sociodemographic or medical characteristics. However, death at home was associated with living together with someone. Advanced hospital-based home care targeting seriously ill cancer patients with a wish to remain at home enable a substantial number of patients to die in the place they desire.
A prospective study of the roles, responsibilities and stresses of chaplains working within a hospice
Authors: Williams Mari Lloyd , Wright Michael , Cobb Mark , Shiels Chris
Published: October 2004

Spiritual care is an integral part of palliative care and if asked, most members of a palliative care team would state they address spiritual issues. The majority of hospices have support from a chaplain. This study was to determine the roles of chaplains within hospices and to look at their levels of stress. A questionnaire containing both open and closed questions was sent to chaplains working within hospices in the UK. The questionnaire enquired about number of sessions, specific roles of chaplain, whether they were members of the multidisciplinary team and their sources of internal support. Stress was measured on a 10-point Likert scale and the GHQ12. One hundred and fifteen questionnaires were returned, with a 72% response rate. The majority (62%) defined their denomination as Church of England and Free Church (24%); 71% of respondents had parish commitments in addition to their hospice role. Roles were predominantly defined as spiritual care of patients and staff (95%) and bereavement support of relatives (76%) and 75% regularly attended the multidisciplinary meetings. Senior medical and nursing staff and other chaplains were perceived as providing most support. Median Likert score for stressfulness was 5, and 23% scored at or above the threshold on the GHQ12 for identifiable psychological morbidity. Clear role definition was associated with less perceived stress whereas the provision of bereavement support was associated with statistically significant increased perceived stress. The role of a chaplain within a hospice is varied and this study suggests that the provision of training and formal support is to be recommended.
Integration of Positive and Negative Affective Stimuli
Authors: Douglas Olsen G. , Pracejus John W.
Published: October 2004

The research reported in this article examines how multiple affective stimuli of differing valence (i.e., both positive and negative) are integrated into an overall affective response. Prior research on the integration of positive and negative information has not focused on affect, and previous affect integration research has not considered both positive and negative feelings. Two experiments investigate how positive and negative affective stimuli combine to influence overall affective responses to an advertisement, content‐specific beliefs, and overall ad evaluations. Both recency and contrast effects were observed on positive affective response and positive beliefs, but comparable effects on negative affective response and negative beliefs were not evident. The results support a motivational influence of affect on judgments.
Akt Phosphorylation Is Not Sufficient for Insulin-Like Growth Factor-Stimulated Myogenin Expression but Must Be Accompanied by Down-Regulation of Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase Phosphorylation
Authors: Tiffin Nicki , Adi Saleh , Stokoe David , Wu Nan-Yan , Rosenthal Stephen M.
Published: October 2004

IGF-I has a unique biphasic effect on skeletal muscle differentiation. Initially, IGF-I inhibits expression of myogenin, a skeletal muscle-specific regulatory factor essential for myogenesis. Subsequently, IGF-I switches to stimulating expression of myogenin. The mechanisms that mediate this switch in IGF action are incompletely understood. Several laboratories have demonstrated that the phosphatidylinositol-3-kinase/Akt signaling pathway is essential for myogenic differentiation and have suggested that this pathway mediates IGF-I stimulation of myogenin mRNA expression, an early critical step in the differentiation process. These studies, however, did not address concurrent Akt and MAPK/ERK1/2 phosphorylation, the latter of which is also known to regulate myogenic differentiation. In the present study in rat L6E9 muscle cells, we have manipulated ERK1/2 phosphorylation with either an upstream inhibitor or activator and examined concurrent levels of Akt and ERK1/2 phosphorylation and of myogenin mRNA expression in response to treatment with IGF-I. We find that even in the presence of phosphorylated Akt, it is only when ERK1/2 phosphorylation is inhibited that IGF-I can stimulate myogenin mRNA expression. Thus, although Akt phosphorylation may be necessary, it is not sufficient for induction of myogenic differentiation by IGF-I and must be accompanied by a decrease in ERK1/2 phosphorylation.
Modification of the Terminal Residue of Apelin-13 Antagonizes Its Hypotensive Action
Authors: Lee Dennis K. , Saldivia Victor R. , Nguyen Tuan , Cheng Regina , George Susan R. , O’Dowd Brian F.
Published: October 2004

The apelin peptide is the endogenous ligand for the apelin G protein-coupled receptor. The distribution of the apelin peptides and receptor are widespread in the central nervous system and periphery, with reported roles in the hypothalamic-pituitary-adrenal axis, blood pressure regulation and as one of the most potent positive inotropic substances yet identified. In this report, we show that in native tissues preproapelin exists as a dimer. Dimeric preproapelin was reduced to monomers by dithiothreitol treatment, indicating disulfide linkages. To evaluate the role of the carboxyl-terminal phenylalanine in the hypotensive action of apelin-13, analogs were generated and tested for their role on blood pressure regulation. Injections of apelin-13 and apelin-12 (15 μg/kg) into spontaneously hypertensive rats lowered systolic and diastolic blood pressure to result in decreases of approximately 60% and 15% in mean arterial blood pressure, respectively. Apelin-13(13[d-Phe]) treatment did not differ from apelin-13 in either efficacy or duration of effect, whereas apelin-13(F13A) revealed a loss of function. However, concomitant administration of apelin-13(F13A) (30 μg/kg) blocked hypotensive effects of apelin-13 (15 μg/kg), which revealed that apelin-13(F13A) behaved as an apelin-specific antagonist.
Stimulation of Steroidogenesis in Immature Rat Leydig Cells Evoked by Interleukin-1α Is Potentiated by Growth Hormone and Insulin-Like Growth Factors
Authors: Colón Eugenia , Svechnikov Konstantin V. , Carlsson-Skwirut Christine , Bang Peter , Soder Olle
Published: October 2004

The cytokine IL-1α is produced constitutively by the intact testis, but its function in this organ remains largely unknown. In this study we examined cooperation between IL-1α and GH and IGFs with regard to stimulation of steroidogenesis by Leydig cells from 40-d-old rats in vitro. IL-1α alone stimulated testosterone (T) and dihydrotestosterone (DHT) production. GH, IGF-I, or IGF-II alone was without effect on T production, but they were found to elevate DHT release, albeit without an obvious dose-response effect. Costimulation with IL-1α and GH or with IL-1α and IGF-I or IGF-II elevated the rate of steroidogenesis (both T and DHT) above that observed with IL-1α alone. GH was found to increase the level of IGF-I in the cultured Leydig cells, an effect that was potentiated by IL-1α. The costimulatory effect of GH on steroidogenesis was abolished by treatment with picropodophyllin, a specific inhibitor of the IGF-I receptor, indicating that the action of GH is mediated via IGF-I. Moreover, cells costimulated with IL-1α and GH exhibited a marked decrease in the level of intact IGF-binding protein-3 in the culture medium due to the induction of proteolytic activity toward this binding protein. In contrast, secretion of IGF-binding protein-2 was increased by such costimulation. These findings suggest that the stimulation of steroidogenesis in Leydig cells evoked by GH and IGFs requires cooperation with IL-1α. This cooperation may play an important role in connection with postnatal Leydig cell maturation and steroidogenesis.
Stimulation of Catecholamine Synthesis by Environmental Estrogenic Pollutants
Authors: Yanagihara Nobuyuki , Toyohira Yumiko , Ueno Susumu , Tsutsui Masato , Utsunomiya Kensuke , Liu Minhui , Tanaka Kazumi
Published: October 2004

Environmental estrogenic pollutants are compounds that have been shown to have estrogenic effects on fetal development and reproductive systems. Less attention, however, has been paid to their influence on neuronal functions. We report here the effects of estrogenic pollutants on catecholamine synthesis in bovine adrenal medullary cells used as a model system of noradrenergic neurons. Treatment of cultured bovine adrenal medullary cells with p-nonylphenol and bisphenol A at 10 nm for 3 d stimulated [14C]catecholamine synthesis from [14C]tyrosine and tyrosine hydroxylase activity, an effect that was not inhibited by ICI 182,780, an antagonist of estrogen receptors. Significant effects of p-nonylphenol on [14C]catecholamine synthesis were observed at 0.1 nm, which is 45 times lower than that of the international regulatory standard (4.5 nm), and the maximum effects were around 10–100 nm. The concentrations (0.1–10 nm) used in the present study are similar to the range observed in rivers in the United States or Europe. On the other hand, short-term treatment of cells with 10 nmp-nonylphenol for 10 min also activated tyrosine hydroxylase, which was suppressed by U0126, an inhibitor of MAPK kinase. Furthermore, treatment of cells with p-nonylphenol for 5 min increased the phospho-p44/42MAPK in a concentration-dependent (1–1000 nm) manner, whereas p-nonylphenol (100 nm, 2 d) enhanced both levels of non-phospho- and phospho-p44/42MAPK. These findings suggest that short-term and long-term treatment of cells with estrogenic pollutants at environmental concentrations stimulates catecholamine synthesis and MAPK through an estrogen receptor-independent pathway.
The 17α and 17β Isomers of Estradiol Both Induce Rapid Spine Synapse Formation in the CA1 Hippocampal Subfield of Ovariectomized Female Rats
Authors: MacLusky Neil J. , Luine Victoria N. , Hajszan Tibor , Leranth Csaba
Published: October 2004

Previous studies have demonstrated that estradiol-17β and estradiol-17α both induce short-latency effects on spatial memory in rats, estradiol-17α being at least as potent as its 17β isomer. To determine whether the mechanisms underlying these behavioral responses might include effects on hippocampal synaptic plasticity, CA1 pyramidal spine synapse density (PSSD) was measured in ovariectomized rats within the first few hours after sc estrogen injection. PSSD increased markedly (by 24%) 4.5 h after the administration of 45 μg/kg estradiol-17β. The PSSD response was significantly greater (44% above control) 30 min after estradiol-17β injection and was markedly dose dependent; a 3-fold lower estradiol-17β dose (15 μg/kg) did not significantly affect CA1 PSSD at either 30 min or 4.5 h. Estradiol-17α was a more potent inducer of PSSD than estradiol-17β. Dose-response analysis determined an ED50 for the effect of estradiol-17α on PSSD of 8.92 ± 1.99 μg/kg, with a maximal response at 15 μg/kg. These results demonstrate that high doses of estradiol induce rapid changes in CA1 PSSD. CA1 spine synapse formation appears to be more sensitive to estradiol-17α than to estradiol-17β, paralleling previous data on the effects of these two steroids on spatial memory. Rapid remodeling of hippocampal synaptic connections may thus contribute to the enhancement of spatial mnemonic processing observed within the first few hours after estrogen treatment. The potency of estradiol-17α suggests that hormone replacement therapy using this steroid might be useful clinically in ameliorating the impact of low endogenous estrogen production on the development and progression of neurodegenerative disorders involving the hippocampus.
Live View of Gonadotropin-Releasing Hormone Containing Neuron Migration
Authors: Bless Elizabeth P. , Walker Heather J. , Yu Kwok W. , Knoll J. Gabriel , Moenter Suzanne M. , Schwarting Gerald A. , Tobet Stuart A.
Published: October 2004

Neurons that synthesize GnRH control the reproductive axis and migrate over long distances and through different environments during development. Prior studies provided strong clues for the types of molecules encountered and movements expected along the migratory route. However, our studies provide the first real-time views of the behavior of GnRH neurons in the context of an in vitro preparation that maintains conditions comparable to those in vivo. The live views provide direct evidence of the changing behavior of GnRH neurons in their different environments, showing that GnRH neurons move with greater frequency and with more changes in direction after they enter the brain. Perturbations of guiding fibers distal to moving GnRH neurons in the nasal compartment influenced movement without detectable changes in the fibers in the immediate vicinity of moving GnRH neurons. This suggests that the use of fibers by GnRH neurons for guidance may entail selective signaling in addition to mechanical guidance. These studies establish a model to evaluate the influences of specific molecules that are important for their migration.
The Orphan Nuclear Receptors NURR1 and NGFI-B Modulate Aromatase Gene Expression in Ovarian Granulosa Cells: A Possible Mechanism for Repression of Aromatase Expression upon Luteinizing Hormone Surge
Authors: Wu Yimin , Ghosh Sagar , Nishi Yoshihiro , Yanase Toshihiko , Nawata Hajime , Hu Yanfen
Published: October 2004

Ovarian granulosa cells play pivotal roles in many aspects of ovary functions including folliculogenesis and steroidogenesis. In response to FSH and LH, the elevation of intracellular cAMP level in granulosa cells leads to activation of multiple ovarian genes. Here, we report findings from a genome-wide study of the cAMP-responsive gene expression profiles in a human granulosa-like tumor cell line, KGN. The study identified 140 genes that are either activated or repressed by 2-fold or greater after stimulation by the adenylyl cyclase activator forskolin. The induction patterns of some cAMP-responsive genes were further analyzed by quantitative real-time PCR. Consistent with previous observations, the LH-responsive genes, such as the nuclear receptor 4A subfamily (NURR1, NGFI-B, and NOR-1), were rapidly but transiently induced, whereas the FSH-responsive gene CYP19 encoding aromatase was induced in a delayed fashion. Interestingly, ectopic expression of NURR1 or NGFI-B severely attenuated the cAMP-responsive activation of the ovary-specific aromatase promoter. Reduction of the endogenous NURR1 or NGFI-B by small interfering RNA significantly elevated aromatase gene expression. The cis-elements responsible for NURR1/NGFI-B-mediated repression were mapped to the minimal aromatase promoter sequence that confers camp responsiveness. Furthermore, the DNA-binding domain of NURR1 was required for the repression. Taken together, these results strongly suggest a causal relationship between the rapid decline of aromatase mRNA and induction of nuclear receptor subfamily 4A expression, which concomitantly occur upon LH surge at the later stages of ovarian follicular development.
The Role of the Locus Coeruleus in Corticotropin-Releasing Hormone and Stress-Induced Suppression of Pulsatile Luteinizing Hormone Secretion in the Female Rat
Authors: Mitchell J. C. , Li X. F. , Breen L. , Thalabard J.-C. , O’Byrne K. T.
Published: October 2004

Despite a wealth of evidence for CRH mediating stress-induced suppression of the hypothalamic GnRH pulse generator, and hence reproductive dysfunction, the site and mechanism of action remains elusive. The locus coeruleus (LC), a prominent noradrenergic brain stem nucleus, is innervated by CRH neurons, mediates several behavioral stress responses, and is implicated in the control of pulsatile LH secretion. The aim of this study was to test the hypothesis that LC CRH has a critical role in mediating stress-induced suppression of pulsatile LH secretion in the rat. Ovariectomized rats with 17β-estradiol or oil-filled sc capsules were implanted with bilateral LC and iv cannulae. Central administration of CRH (10 ng to 1 μg) resulted in a dose-dependent suppression of LH pulses, which was reversed by a CRH receptor antagonist (α-helical CRF9–41, 1 μg). The induction of c-fos expression in glutamic acid decarboxylase67 immunostained neurons in the preoptic area suggests activation of the secretion of γ-aminobutyric acid in response to intracoerulear administration of CRH; 17β-estradiol further increased the percentage of glutamic acid decarboxylase67-positive neurons that expressed fos and augmented suppression of LH pulses. Furthermore, intracoerulear administration of α-helical CRF9–41 completely blocked restraint stress-induced suppression of LH pulses, without affecting the inhibitory response to hypoglycemia. These results suggest that CRH innervation of the LC may play a pivotal, but differential, role in the normal physiological response of stress-induced suppression of the GnRH pulse generator and hence the reproductive system.
Glucagon-Like Peptide 2 Dose-Dependently Activates Intestinal Cell Survival and Proliferation in Neonatal Piglets
Authors: Burrin Douglas G. , Stoll Barbara , Guan Xinfu , Cui Liwei , Chang Xiaoyan , Holst Jens J.
Published: October 2004

Glucagon-like peptide 2 (GLP-2) is a gut hormone that stimulates mucosal growth in total parenteral nutrition (TPN)-fed piglets; however, the dose-dependent effects on apoptosis, cell proliferation, and protein synthesis are unknown. We studied 38 TPN-fed neonatal piglets infused iv with either saline or GLP-2 at three rates (2.5, 5.0, and 10.0 nmol·kg–1·d−1) for 7 d. Plasma GLP-2 concentrations ranged from 177 ± 27 to 692 ± 85 pm in the low- and high-infusion groups, respectively. GLP-2 infusion dose-dependently increased small intestinal weight, DNA and protein content, and villus height; however, stomach protein synthesis was decreased by GLP-2. Intestinal crypt and villus apoptosis decreased and crypt cell number increased linearly with GLP-2 infusion rates, whereas cell proliferation and protein synthesis were stimulated only at the high GLP-2 dose. The intestinal activities of caspase-3 and -6 and active caspase-3 abundance decreased, yet procaspase-3 abundance increased markedly with increasing infusion rate and plasma concentration of GLP-2. The GLP-2-dose-dependent suppression of intestinal apoptosis and caspase-3 activity was associated with increased protein kinase B and glycogen-synthase kinase-3 phosphorylation, yet the expression phosphatidylinositol 3-kinase was unaffected by GLP-2. Intestinal endothelial nitric oxide synthase mRNA and protein expression was increased, but only at the high GLP-2 dose. We conclude that the stimulation of intestinal epithelial survival is concentration dependent at physiological GLP-2 concentrations; however, induction of cell proliferation and protein synthesis is a pharmacological response. Moreover, we show that GLP-2 stimulates intestinal cell survival and proliferation in association with induction of protein kinase B and glycogen-synthase kinase-3 phosphorylation and Bcl-2 expression.
Intestinal Lipoprotein Overproduction, a Newly Recognized Component of Insulin Resistance, Is Ameliorated by the Insulin Sensitizer Rosiglitazone: Studies in the Fructose-Fed Syrian Golden Hamster
Authors: Lewis Gary F. , Uffelman Kristine , Naples Mark , Szeto Linda , Haidari Mehran , Adeli Khosrow
Published: October 2004

We investigated whether intestinal lipoprotein overproduction in a fructose-fed, insulin-resistant hamster model is prevented with insulin sensitization. Syrian Golden hamsters were fed either chow, 60% fructose for 5 wk, chow for 5 wk with the insulin sensitizer rosiglitazone added for the last 3 wk, or 60% fructose plus rosiglitazone. In vivo Triton studies showed a 2- to 3-fold increase in the large (Svedberg unit > 400) and smaller (Sf 100–400) triglyceride-rich lipoprotein particle apolipoprotein B48 (apoB48) but not triglyceride secretion with fructose feeding in the fasted state (P < 0.01) and partial normalization with rosiglitazone in fructose-fed hamsters. Ex vivo pulse-chase labeling of enterocytes confirmed the oversecretion of apoB48 lipoproteins with fructose feeding. Intestinal lipoprotein oversecretion was associated with increased expression of microsomal triglyceride transfer protein expression. With rosiglitazone treatment of fructose-fed hamsters, there was approximately 50% reduction in apoB48 secretion from primary cultured enterocytes and amelioration of the elevated microsomal triglyceride transfer protein mass and activity in fructose-fed hamsters. In contrast, in the postprandial state, the major differences between nutritional and drug intervention protocols were evident in triglyceride-rich lipoprotein triglyceride and not apoB48 secretion rates. The data suggest that intestinal lipoprotein overproduction can be ameliorated with the insulin sensitizer rosiglitazone.
Internal ribosome entry site drives cap-independent translation of reaper and heat shock protein 70 mRNAs in Drosophila embryos
Authors: HERNÁNDEZ GRECO , VÁZQUEZ-PIANZOLA PAULA , SIERRA JOSÉ M. , RIVERA-POMAR ROLANDO
Published: October 2004

Translation is a sensitive regulatory step during cellular stress and the apoptosis response. Under such conditions, cap-dependent translation is reduced and internal ribosome entry site (IRES)-dependent translation plays a major role. However, many aspects of how mRNAs are translated under stress remain to be elucidated. Here we report that reaper mRNA, a pro-apoptotic gene from Drosophila melanogaster, is translated in a cap-independent manner. In Drosophila mutant embryos devoid of the eukaryotic initiation factor 4E (eIF4E), reaper transcription is induced and apoptosis proceeds. In vitro translation experiments using wild-type and eIF4E mutant embryonic extracts show that reporter mRNA bearing reaper 5′ untranslated region (UTR) is effectively translated in a cap-independent manner. The 5′UTR of reaper exhibits a high degree of similarity with that of Drosophila heat shock protein 70 mRNA, and both display IRES activity. Studies of mRNA association to polysomes in embryos indicate that both reaper and heat shock protein 70 mRNAs are recruited to polysomes under apoptosis or thermal stress. Our data suggest that heat shock protein 70 and reaper, two antagonizing factors in apoptosis, use a similar mechanism for protein synthesis.
Adjunctive Granulocyte Colony-Stimulating Factor Therapy for Diabetic Foot Infections
Authors: Reed Kelly S , Pai Manjunath P
Published: October 2004

OBJECTIVE

To evaluate the role of granulocyte colony-stimulating factor (G-CSF) as adjunctive therapy for the treatment of diabetic foot infections in non-neutropenic patients.

DATA SOURCES

Clinical literature was accessed through MEDLINE (1965–April 2004). Key search terms included G-CSF, infection, and diabetes. In addition, relevant references from primary and secondary article bibliographies were extracted.

DATA SYNTHESIS

Three clinical trials evaluating G-CSF for diabetic foot infections were identified. These data demonstrated positive effects of G-CSF on improvement of foot infections and risk of amputations.

CONCLUSIONS

Controlled trials are necessary to validate the role of adjunctive G-CSF at reducing amputations in patients with diabetic foot infections.
Trivalent Intranasal Influenza Vaccine, Live
Authors: McCarthy Michelle W , Kockler Denise R
Published: October 2004

OBJECTIVE

To review human data on the efficacy, safety, and clinical use of trivalent intranasal influenza vaccine, live (TIIVL).

DATA SOURCES

A MEDLINE search (1966–3rd week of January 2004) using the terms influenza vaccine, intranasal administration, and FluMist was conducted. References from pertinent articles were also reviewed.

STUDY SELECTION AND DATA EXTRACTION

Studies conducted in humans and published in English were selected. Double-blind, controlled trials evaluating the efficacy and safety of TIIVL were evaluated.

DATA SYNTHESIS

Administration of TIIVL results in mucosal and humoral immunity to influenza. Results of clinical trials in children and adults have demonstrated that TIIVL reduces the incidence of influenza. In children, TIIVL was also associated with a decrease in febrile illness and febrile otitis media. In adults, reductions in workday absences and medical visits due to febrile upper respiratory tract illness were also documented. TIIVL is well tolerated, with rhinorrhea or nasal congestion and sore throat occurring more frequently than with placebo.

CONCLUSIONS

TIIVL is an alternative to intramuscular inactivated influenza vaccine in healthy individuals between 5 and 49 years of age. However, the vaccine is contraindicated in the majority of patient populations for whom annual influenza vaccination is recommended. THIS ARTICLE IS APPROVED FOR CONTINUING EDUCATION CREDIT ACPE UNIVERSAL PROGRAM NUMBER: 407-000-04-037-H01
Exocyclic DNA Adducts as Oxidative Stress Markers in Colon Carcinogenesis: Potential Role of Lipid Peroxidation, Dietary Fat and Antioxidants
Authors: Bartsch Helmut , Nair Jagadeesan , Owen Robert W.
Published: October 2004

Molecular pathways to colorectal cancer involve multiple genetic changes, whereby extensive oxyradical damage causes mutations in cancerrelated genes and leads to a cycle of cell death and regeneration. Besides direct oxidative DNAdamage, reactive oxygen and nitrogen species can induce etheno ()DNA adducts mainly via trans-4-hydroxy-2-nonenal, generated as the major aldehyde by lipid peroxidation (LPO) of ω-6 PUFAs. Patients with familial adenomatous polyposis (FAP) develop multiple colorectal adenomas. In affected tissues increased LPO could be triggered due to increased arachidonic acid metabolism as a result of elevated cyclooxygenases. Our studies demonstrated an increased DNA adduct level in affected colon epithelia of FAP patients. DNA adducts are promutagenic and can cause genomic instability that drives colorectal adenoma to malignancy. We have further investigated the potential chemopreventive properties of olive oil and its polyphenolic components. Mediterranean diet, of which olive oil is a major fatty acid source, has protective effects against human breast and colorectal cancers. Olive oil extracts and the newly identified lignan fractions showed high antioxidant capacity in vitro. As DNA adducts are biomarkers for oxidative stress and LPO induced DNA damage, they can verify the efficacy of newly identified antioxidants, e.g. from olive oil, as chemopreventive agents against colon carcinogenesis.
Functional Genomics of C190T Single Nucleotide Polymorphism in Human N-Acetyltransferase 2
Authors: Zhu Yuanqi , Doll Mark A. , Hein David W.
Published: October 2004

Nacetyltransferase 2 (NAT2) catalyzes Nacetylation and Oacetylation of many drugs and environmental carcinogens. Genetic polymorphisms in the NAT2 gene have been associated with differential susceptibility to cancers and drug toxicity from these compounds. Single nucleotide polymorphisms (SNPs) have been identified in the human NAT2 coding region. A new allele, NAT2*19, possessing the C190T (R64W) exchange, was recently identified. In order to understand the effect of this new SNP, recombinant NAT2*4 (reference) and NAT2*19 were expressed in yeast (Schizosaccharomyces pombe). The C190T (R64W) SNP in NAT2*19 caused substantial reduction in the NAT2 protein level and stability, but did not cause significant reduction in transformation efficiency or mRNA level. The enzymatic activities for Nacetylation of two arylamine carcinogens (2-aminofluorene, 4-aminobiphenyl), and a sulfonamide drug (sulfamethazine) were over 100-fold lower for NAT2 19 compared to reference NAT2 4. Kinetic studies showed a reduction in Vmax but no significant change in substrate Km. In addition, the SNP caused significant reduction in the Oacetylation of the Nhydroxy 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine. These results show that NAT2*19 possessing the C190T (R64W) SNP encodes a slow acetylator phenotype for both N and Oacetylation, due to a reduction in the amount and stability of the NAT2 19 allozyme.
Specificity Determinants in the Interaction of Apolipoprotein(a) Kringles with Tetranectin and LDL
Authors: Caterer N. R. , Graversen J. H. , Jacobsen C. , Moestrup S. K. , Sigurskjold B. W. , Etzerodt M. , Thøgersen H.C.
Published: October 2004

Lipoprotein(a) is composed of low density lipoprotein and apolipoprotein(a). Apolipoprotein(a) has evolved from plasminogen and contains 10 different plasminogen kringle 4 homologous domains [KIV(1 110)]. Previous studies indicated that lipoprotein(a) noncovalently binds the Nterminal region of lipoprotein B100 and the plasminogen kringle 4 binding plasma protein tetranectin. In this study recombinant KIV(2), KIV(7) and KIV(10) derived from apolipoprotein(a) were produced in E. coli and the binding to tetranectin and low density lipoprotein was examined. Only KIV(10) bound to tetranectin and binding was similar to that of plasminogen kringle 4 to tetranectin. Only KIV(7) bound to LDL. In order to identify the residues responsible for the difference in specificity between KIV(7) and KIV(10), a number of surfaceexposed residues located around the lysine binding clefts were exchanged. Ligand binding analysis of these derivatives showed that Y62, and to a minor extent W32 and E56, of KIV(7) are important for LDL binding to KIV(7), whereas R32 and D56 of KIV(10) are required for tetranectin binding of KIV(10).
The Primary Structure of Three Hemoglobin Chains from the Indigo Snake (Drymarchon corais erebennus, Serpentes): First Evidence for αD Chains and Two β Chain Types in Snakes
Authors: Stoeckelhuber M. , Gorr T. , Kleinschmidt T.
Published: October 2004

The hemoglobin of the indigo snake (Drymarchon corais erebennus, Colubrinae) consists of two components, HbA and HbD, in the ratio of 1:1. They differ in both their α and β chains. The amino acid sequences of both α chains (αA and αD) and one β chain (βI) were determined. The presence of an αDchain in a snake hemoglobin is described for the first time. A comparison of all snake β chain sequences revealed the existence of two paralogous β chain types in snakes as well, which are designated as βI and βII type. For the discussion of the physiological properties of Drymarchon hemoglobin, the sequences were compared with those of the human α and β chains and those of the closely related water snake Liophis miliaris where functional data are available. Among the heme contacts, the substitution αD58(E7)His->Gln is unusual but most likely without any effect. The residues responsible for the main part of the Bohr effect are the same as in mammalian hemoglobins. In each of the three globin chains only two residues at positions involved in the α1/_2 interface contacts, most important for the stability and the properties of the hemoglobin molecule, are substituted with regard to human hemoglobin. On the contrary, nine, eleven, and six α1/β1 contact residues are replaced in the αA, αD, βI chains, respectively.
Identification and Characterization of an IgG Binding Protein in the Secretion of the Rat Coagulating Gland
Authors: Wilhelm B. , Keppler C. , Henkeler A. , Schilli-Westermann M. , Linder D. , Aumüller G. , Seitz J.
Published: October 2004

A merocrine released protein (named 115k protein) was highly enriched from the secretion of the rat coagulating gland. The protein has a molecular mass of 115 kDa as calculated by SDSPAGE under reducing conditions. Furthermore, the 115 kDa protein is glycosylated, and carries Man, GlcNAc, Gal, Fuc and sialic acid residues. For identification, Nterminal amino acid and nucleotide sequence analyses were performed. The sequences obtained showed 86 to 100% identity with human and mouse IgGFc binding proteins. The functional capacity of IgG binding of the 115 kDa protein was shown by overlay experiments, indicating its membership in the IgG binding protein family.
X-ray structure of fumarylacetoacetate hydrolase family member Homo sapiens FLJ36880
Authors: Manjasetty Babu A. , Niesen Frank H. , Delbrück Heinrich , Götz Frank , Sievert Volker , Büssow Konrad , Behlke Joachim , Heinemann Udo
Published: October 2004

The human protein FLJ36880 belongs to the fumarylacetoacetate hydrolase family. The X-ray structure of FLJ36880 has been determined to 2.2 Å resolution employing the semi-automated high-throughput structural genomics approach of the Protein Structure Factory. FLJ36880 adopts a mixed β-sandwich roll fold and forms homodimers in crystals as well as in solution. One Mg2+ ion is bound to each subunit of the dimeric protein by coordination to three carboxylate oxygens and three water molecules. These metal binding sites are accessible from the same surface of the dimer, partly due to the disorder of the undecapeptide stretch D29 to L39. The overall structure and metal binding site of FLJ36880 bear clear similarities to the C-terminal domain of the bifunctional enzyme HpcE from Escherichia coli C, fumarylacetoacetate hydrolase from Mus musculus and to YcgM (Apc5008) from E. coli 1262. These similarities provide a framework for suggesting biochemical functions and evolutionary relationships of FLJ36880. It appears highly probable that the metal binding sites are involved in an enzymatic activity related to the catabolism of aromatic amino acids. Two point mutations in the active-site of FAH, responsible for the metabolic disease hereditary tyrosinemia type I (HTI) in humans, affect residues that are structurally conserved in FLJ36880 and located in the putative catalytic site.
Segregation of partly melted molecules: isolation of CpG islands by polyacrylamide gel electrophoresis
Authors: Shiraishi Masahiko , Oates Adam J. , Li Xu , Chuu Ying H. , Sekiya Takao
Published: October 2004

The technique of segregation of partly melted molecules (SPM) is a convenient and efficient method to isolate DNA fragments associated with CpG islands. The approach is conceptually simple and uses denaturant gradient gel electrophoresis to separate DNA molecules digested with restriction endonucleases. The SPM methodology has successfully been applied to the identification of genes from anonymous, unsequenced DNA fragments and CpG islands methylated in human cancer. In this article the theoretical background and practical application of the SPM method is reviewed.
Measurement of Water Turnover Using Deuterium Dilution: Impact of Periodically Varying Turnover Rates on Precision and Accuracy
Authors: Fusch Christoph
Published: October 2004

Water metabolism may be non-invasively investigated
using deuterium elimination from the body. Evaluation
of elimination constants (kD) and pool sizes (N) becomes
complicated when turnover rates vary periodically - a realistic scenario in free-living subjects.
Unfortunately, phase and frequency of periodic variations
are not a priori known and can therefore not be
used for a sampling protocol. This study investigates
the impact of periodical flux variations on the measurement
of water turnover. For two models of periodic
variation with identical apparent kD, data sets of
tracer concentrations in body water were generated
for two-hour intervals using a monoexponential decay
equation. Data were analyzed using the two- and the
multi-point method increasing observation periods
stepwise. Apparent values for kD, N and water turnover
(RH₂O) were compared with values originally used
in the model. Periodically varying turnover rates introduce
considerable errors for kD and RH₂O when the two-point
method is used. The multi-point method gives a
more robust estimate of RH₂O already after short observation
periods, however, due to slope-intercept
correlations, still tends to overestimate RH₂O. Errors
are more pronounced the more uneven flux rates are
distributed. Stepwise analysis of tracer enrichments
using the multi-point approach identifies periodical
variation of flux rates.
Evaluating Agreement between Two Analytical Methods in Clinical Chemistry
Authors: Magari Robert T.
Published: October 2004

A test and a reference analytical method are usually
compared for agreement based on paired data obtained
from several independent subjects. Bias between
two methods can be classified as constant and
proportional. Bias is modeled as an average bias between
two methods (constant bias) and proportional
bias related to individual measurements. A weighted
least square approach is followed for estimating the
parameters. Estimates of constant and proportional
biases are tested individually and their significance
can be used to explain the sources of disagreement between
two methods and help deciding a remedial
strategy. The proposed model can also be used to determine
an optimum combination of subjects and runs
per subject and facilitate the allocation of the resources.
Evaluation of Two Automated Enzyme Immunoassays for Detection of Antinuclear Antibodies
Authors: Bossuyt Xavier
Published: October 2004

We evaluated two enzyme immunoassays (EIA) for detection
of antinuclear antibodies (ANA) which became
recently available and which were designed for application
on a fully automated system; (i) the EIA-ANA
screen kit from Sigma Diagnostics (St. Louis, MO, USA)
applied on APTUS™, an automated EIA analyser from
Sigma Diagnostics and (ii) the Cobas® Core Hep-2 EIA-ANA
assay applied on the fully automated Cobas Core
immunochemistry analyzer from Roche Diagnostics
(Basel, Switzerland). The evaluation was done by an
analytic comparison of the automated systems to an
established indirect immunofluorescence (IF) method
performed on SSA transfected human epitheloid cell
substrate slides. Three hundred and thirty six samples
were tested with the Sigma EIA-ANA assay and 603
samples with the Cobas® Core Hep-2 EIA-ANA assay.
For both EIA systems, there was a trend of generally
increasing signal from the assay system with increasing
IF-ANA titers. With an IF-ANA reference range of
< 1:160, concordance between Sigma® EIA-ANA and IFANA
was 86% and concordance between Roche® EIA-ANA and IF-ANA was 85%. When compared to IF-ANA
with a reference range of < 1:160, sensitivity and specificity
of the EIA-ANA screen was 0.65 and 0.92, respectively,
for Sigma® EIA-ANA and 0.61 and 0.91, respectively,
for the Cobas® Core assay. The low sensitivity
observed with both methods is a major concern. Sigma
EIA-ANA screen revealed the presence of autoantibodies
in 23 of the 29 samples containing antibodies to extractable
nuclear antigens (ENA) and/or double
stranded DNA and the Cobas® Core Hep-2 EIA-ANA assay
revealed the presence of autoantibodies in 40 of the
43 samples containing antibodies to ENA and/or double
stranded DNA.
Performance Evaluation of Nine Hormone Assays on the Immulite 2000®Immunoassay System
Authors: Tello Francisca Llinares , Hernández Dolores Mirete
Published: October 2004

We evaluated the analytical performance of the Immulite
2000® immunoassay analyzer (Diagnostic Products
Corporation, Los Angeles, USA) based on a new
detection technology, electrochemical luminescence.
The evaluated analytes were thyrotropin, triiodothyronine,
free thyroxine, follitropin, lutropin, prolactin, cortisol,
estradiol and progesterone. We tested the assay
precision, linearity, recovery, and correlation with
comparison methods for these analytes. For most assays,
within-run and between-day imprecisions were
less than 8% and 10%, respectively. The linearity and
recovery were acceptable for all assays. The correlation
between the Immulite 2000® assays and comparison
methods showed satisfactory results.
Usefulness of EC4 Essential Criteria for Quality Systems of Medical Laboratories as Guideline to the ISO 15189 and ISO 17025 Documents
Authors: T.P.Jansen Rob , Kenny Desmond , Blaton Vic , Burnett David , Huisman Wim , Plebani Mario , Queraltó Josep M. , Zérah Simone , Lieshout Joep van
Published: October 2004

Many medical laboratories have made a start with the
introduction of quality management systems. However,
it is still not clear against which standards such
systems should be measured. The existing ISO and
CEN standards do not cover essential aspects of medical
laboratories. The publication of the EC4 Essential
Criteria has stimulated the development of the
ISO/Draft International Standard 15189. This standard
seems adequate for our type of laboratories. However,
it is not easy to read. The EC4 Essential Criteria could
well serve as a guide, covering additional aspects, e.g.
on total quality management and budget management
as required in the EFQM model, that are not (yet)
included in the ISO standard. In the present article the
EC4 Essential Criteria are cross-referenced with two
new international ISO standards, ISO/FDIS 15189 and
ISO/FDIS 17025, the latter being the successor of ISO
guide 25 and EN 45000. Both new ISO documents are
in compliance with the new ISO 9000:2000 standard.
Recommendations for Measurement of and Conventions for Reporting Sodium and Potassium by Ion-Selective Electrodes in Undiluted Serum, Plasma or Whole Blood
Authors: Burnett Robert W. , Covington Arthur K. , Fogh-Andersen Niels , Külpmann Wolf R. , Lewenstam Andrzej , Maas Anton H.J. , Müller-Plathe Oswald , Sachs Charles , Siggaard-Andersen Ole , VanKessel Antonius L. , Zijlstra Willem G.
Published: October 2004

Ion-selective electrodes (ISEs) respond to ion-activity
and therefore do not sense substance concentration directly. However, it is recognized that sodium and
potassium in plasma will continue to be expressed for
clinical purposes in terms of substance concentration
(mmol/l). A convention is proposed whereby for routine
clinical purposes results of ISE measurements
of sodium and potassium in undiluted plasma should
be reported in terms of substance concentration
(mmol/l). In specimens with normal concentrations of
plasma water, total CO2, lipids, protein and pH, the values
will concur with the total substance concentration
as determined for example by flame atomic emission
spectrometry (FAES) or ISE measurements on diluted
samples. In specimens with abnormal concentrations
of plasma water, the results will differ. However, under
these circumstances, measurements of sodium and
potassium by ISE in the undiluted sample will more
appropriately reflect the activity of sodium and potassium
and are therefore clinically more relevant than
the determination in diluted samples. Detailed recommendations
are made about practical procedures to
achieve this. The recommended name for this quantity
is the substance concentration of ionized sodium or
ionized potassium in plasma, as opposed to total
sodium or total potassium determined by, e.g. FAES,
or ISE measurements on diluted samples.