bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in Family Structure After a Renal Transplant
Authors: DE PARRA MARÍA LUISA VELASCO
Published: August 2004

Two girls (9 and 10 years old) and one male adolescent (19 years old) with chronic renal failure (CRF) were studied along with their families in order to evaluate their family structure before (4–5 weeks) and after (3–5 weeks) a renal transplant from a live donor. After the renal transplant there was a close association between the donor and the recipient that led to the creation of an alliance or the reinforcement of an alliance already present before the transplant. This may favor a family disruption or a better family equilibrium depending upon whether the donor ranked highest in the family hierarchy and was already in a rigid alliance with the patient before the transplant. It is suggested that the study and diagnosis of the family structure before the renal transplant may help to: (a) select the most suitable donor candidate from a biological as well as psychological point of view so as to allow a better possibility of success in the structural functioning of the family after the transplant; (b) predict changes that will appear within the family after the transplant, which in turn may alert the personnel to detect changes in the family structure as early as possible; and (c) offer more rapid therapeutic assistance to bring the family back to a functional level within its social framework.
Helping Mom Fail: A Case of a Stalemated Divorcing Process
Authors: ISAACS MARLA BETH
Published: August 2004

This article describes a systems approach to the problem of a child who had become symptomatic as a result of her parents' separation. Elements of the divorcing process that can create a symptomatic child are highlighted. Family members cope with separation and divorce by devising strategies for navigating the divorcing process. These strategies can mesh to create dysfunctional sequences in which family members get “stuck.” Case material illustrates the development of a stalemated divorcing process, in which a dysfunctional sequence keeps the child uncontrollable and mother incompetent. Family therapy breaks the stalemate and allows family members to proceed with appropriate developmental tasks.
On the Problems of Eclecticism: A Call for Epistemologic Clarification and Human‐Scale Theories
Authors: LIDDLE HOWARD ARTHUR
Published: August 2004

This paper addresses several contemporary theoretical issues in the family therapy field: clinicians' and students' selection of a theoretical orientation in light of the political struggles of competing schools of thought; problems of eclecticism for the practicing family therapist; the role of theory in clinical practice; and size and scope issues in theory‐building. Finally, the paper suggests a periodic activity, termed an Epistemologic Declaration, designed to help therapists clarify and refine their basic therapeutic beliefs.
Circular Questioning
Authors: PENN PEGGY
Published: August 2004

The plan of this paper is to explore the question: Does a model that includes the principles of double description, circularity, and coevolutionary change, all accounting for shifts in family coalitions over time and the emergence of problems in connection with these shifts, allow the family therapist to design better methods for the understanding and practice of family therapy?

Concepts of double description, coevolution, and circularity from Gregory Bateson's writing and the research of other scientists describe the translation of these ideas from pure epistemology to the pragmatics of family therapy. Circular questioning developed by the Milan Associates is presented as a practice method exemplifying how these notions of circularity and coevolutionary change — especially changes in family patterns — are used during actual family sessions.
A Case of the Blind Leading the “Blind”: Reframing a Physical Handicap as Competence
Authors: WEBB‐WOODARD LINDA , WOODARD BRENT
Published: August 2004

Structural and strategic family therapy inform the course of treatment of a family with a blind member. Alteration of the family's view of the problem through reframing the blindness as competence is reported.
A Note on Spinoza's Contribution to Systemic Therapy
Authors: KRAEMER SEBASTIAN
Published: August 2004

Writing 300 years ago the philosopher Spinoza anticipated some of the fundamental principles of systemic therapy, notably a respect for the remorseless logic of natural systems and the value of positive connotation. In the debate about the ethics of paradoxical methods, he offers a logical, and humble, defense.
The Therapeutic Advantage to the Family Therapist of Intervening in the Family‐Clinician System
Authors: HURLEY R. PETER
Published: August 2004

Attempting family therapy in the context of a family‐clinician system poses extreme difficulties for the therapist. He cannot be effective if he diagnoses only the family system. He must broaden his analysis to include other clinicians' transactions with the family. Once he accomplishes this perceptual leap, he can intervene in the family‐clinician system most effectively with paradoxical methods. He may find that a change in his own contribution to the system solves the family's problem.
Energy use in spawning Atlantic salmon
Authors: Hendry A. P. , Beall E.
Published: August 2004

Abstract –  We studied some of the factors that might influence energy use in spawning Atlantic salmon (Salmo salar). Single females were placed into an experimental channel with either one or three males, after which spawning was monitored continuously. Male status was confirmed using genetic parentage analysis. Daily fat loss was monitored with the Torry Fish Fatmeter and validated through biochemical analyses. Several comparisons were in the expected direction but not statistically significant and therefore require further study: daily fat loss appeared higher for dominant males relative to subordinate males and in the three‐male treatment relative to the one‐male treatment. Most of the variation among individuals remained unexplained, suggesting that several as yet unknown factors strongly influence fat loss in spawning salmon. A large and significant effect was that daily fat loss was higher for females than for males, a difference that might contribute to the shorter spawning duration typical of females.
Sex‐specific growth pattern during early life history in masu salmon, Oncorhynchus masou
Authors: Yamamoto T.
Published: August 2004

Abstract –  To examine whether or not sex‐specific growth occurs in fry of masu salmon Oncorhynchus masou, 90 half‐sibling fry chosen from three different classes of egg size (small, medium and large) and emergence time (period from fertilisation to first feeding; early, middle and late) were raised in a tank for 60 days. In all four replications, male fry grew faster than females. The difference in growth between sexes was greater among siblings hatched from larger eggs. Fast‐growing, young male masu salmon show a precocious form that tends to mature without migrating to sea. Slower growing males tend to show the anadromous form, which is only shown in females. Sex‐specific life histories may influence the sex‐specific growth patterns during early life history.
Effects of constant and cyclical thermal regimes on growth and feeding of juvenile cutthroat trout of variable sizes
Authors: Meeuwig M. H. , Dunham J. B. , Hayes J. P. , Vinyard G. L.
Published: August 2004

Abstract –  The effects of constant (12, 18, and 24 °C) and cyclical (daily variation of 15–21 and 12–24 °C) thermal regimes on the growth and feeding of Lahontan cutthroat trout (Oncorhynchus clarki henshawi) of variable sizes were examined. Higher constant temperatures (i.e., 24 °C) and more variable daily temperatures (i.e., 12–24 °C daily cycle) negatively affected growth rates. As fish mass increased (from 0.24 to 15.52 g) the effects of different thermal regimes on mass growth became more pronounced. Following 14 days exposure to the thermal regimes, feeding rates of individual fish were assessed during acute exposure (40 min) to test temperatures of 12, 18, and 24 °C. Feeding rate was depressed during acute exposure to 24 °C, but was not significantly affected by the preceding thermal regime. Our results indicate that even brief daily exposure to higher temperatures (e.g., 24 °C) can have considerable sublethal effects on cutthroat trout, and that fish size should be considered when examining the effects of temperature.
Effect of inhibiting melatonin biosynthesis on spatial memory retention and tau phosphorylation in rat
Authors: Zhu Ling Qiang , Wang Shao Hui , Ling Zhi Qun , Wang Dan Ling , Wang Jian‐Zhi
Published: August 2004

Abstract:  We have found recently that melatonin protects SH‐SY5Y neuroblastoma cells from calyculin A‐induced neurofilament impairment and neurotoxicity. In the present study, we further investigated the in vivo effect of inhibiting melatonin biosynthesis on spatial memory retention and tau phosphorylation in rats and the potential underlying mechanisms by using haloperidol, a specific inhibitor of 5‐hydroxyindole‐O‐methyltransferase, and a key enzyme in melatonin biosynthesis. We have found that injection of haloperidol into the lateral ventricle and into peritoneal cavity compromises spatial memory retention of rats and induces hyperphosphorylation of microtubule‐associated protein tau at tau‐1 (Ser199/Ser202) and PHF‐1 (Ser396/Ser404) epitopes. At mean time, the activity of protein phosphatase‐2A (PP‐2A), a deficit phosphatase in the Alzheimer's disease brain and superoxide dismutase decreases with an elevated level of malondialdehyde. Supplementation with melatonin by prior injection for 1 wk and reinforcement during the haloperidol administration significantly improves memory retention deficits, arrests tau hyperphosphorylation and oxidative stress, and restores PP‐2A activity. These results strongly support the involvement of decreased melatonin in Alzheimer‐like spatial memory impairment and tau hyperphosphorylation, and PP‐2A may play a role in mediating aberrant melatonin‐induced lesions.
Melatonin stimulates calmodulin phosphorylation by protein kinase C
Authors: Soto‐Vega Elena , Meza Isaura , Ramírez‐Rodríguez Gerardo , Benitez‐King Gloria
Published: August 2004

Abstract:  Calmodulin (CaM)‐dependent processes can be modulated by the availability of Ca+2, the subcellular distribution of both CaM and its target proteins, CaM antagonism, and post‐translational modifications such as CaM phosphorylation. Melatonin, the pineal secretory product synthesized during the dark phase of the photoperiod is an endogenous CaM antagonist. This indolamine causes CaM subcellular redistribution in epithelial MDCK and MCF‐7 cells, and selectively activates protein kinase C alpha (PKC α) in neuronal N1E‐115 cells. In the present work we have characterized the phosphorylation of CaM mediated by PKC α and its stimulation by melatonin in an in vitro reconstituted enzyme system. Additionally, the participation of MAPK and ERKs, downstream kinases of the PKC signaling pathway, was explored utilizing MDCK cell extracts as source of these kinases. Phosphorylation of CaM was characterized in the whole cells by MDCK cell metabolic labeling with [32P]‐orthoposhospate, and CaM separation by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis, as well as by immunocolocalization of phosphorylated threonine/serine residues and CaM in cultured cells incubated with melatonin. Our results show that melatonin increased CaM phosphorylation by PKC α with an EC50 of 10−8 m in the presence of the phorbol ester, phorbol‐12‐myristate‐13‐acetate (PMA) in the in vitro reconstituted enzyme system. An increase in phosphorylated CaM was also observed in cells cultured with melatonin, or PMA for 2 hr, while, PKC, MAPK, or ERK inhibitors abolished CaM phosphorylation elicited by melatonin in MDCK cell extracts. Our data show that melatonin can stimulate phosphorylation of CaM by PKC α in the in vitro reconstituted system and suggest that in MDCK cells this phosphorylation is accomplished by PKC. Modification of CaM by melatonin can be another route to inhibit CaM interaction with its target enzymes.
Melatonin reduces torsion–detorsion injury in rat ovary: biochemical and histopathologic evaluation
Authors: Turkoz Yusuf , Celik Onder , Hascalik Seyma , Cigremis Yılmaz , Hascalik Mehmet , Mizrak Bulent , Yologlu Saim
Published: August 2004

Abstract:  This experimental study was designed to determine the effects of melatonin on the levels of malondialdehyde (MDA), reduced glutathione (GSH), xanthine oxidase (XO) after adnexial torsion/detorsion (ischemia/reperfusion, I/R) of the ovaries of in rats. Forty adult albino rats were divided into five groups: sham operation, torsion, I/R plus saline, I/R plus melatonin and torsion plus melatonin. Rats in the sham‐operated group underwent a surgical procedure similar to the other groups but the adnexa was not occluded. Rats in the torsion group were killed after adnexal torsion for 3 hr. Melatonin and saline were injected intraperitoneally (10 mg/kg) 30 min before detorsion to the I/R plus melatonin group and I/R plus saline group respectively. After 3 hr of ovarian detorsion, the rats were killed and ovaries were removed. Melatonin was injected intraperitoneally (10 mg/kg) 30 min before torsion to the torsion plus melatonin group. After 3 hr of ovarian torsion, the rats were killed and ovaries were harvested. The tissue levels of MDA, GSH and XO were measured. MDA and XO levels in the I/R plus saline group increased significantly when compared with torsion and sham‐operated groups (P < 0.001). MDA and XO levels in the I/R plus melatonin group were lower than I/R plus saline and differences between the two groups were statistically significant (P < 0.001). GSH levels in the I/R plus saline group decreased significantly when compared with ischemia and sham‐operated groups (P < 0.001). GSH levels in the I/R plus melatonin treated rats were significantly higher than I/R plus saline and ischemia groups (P < 0.001). The tissue levels of XO, MDA and GSH were similar between ischemia and ischemia plus melatonin groups. Morphologically, polymorphonuclear neutrophil infiltration and vascular dilatation were obvious in the I/R‐damaged ovaries, and the changes also partially reversed by melatonin. This study demonstrates that melatonin protects the ovaries against oxidative damage associated with reperfusion following an ischemic insult.
The Semaphorin 4D Receptor Plexin-B1 Is a GTPase Activating Protein for R-Ras
Authors: Oinuma Izumi , Ishikawa Yukio , Katoh Hironori , Negishi Manabu
Published: August 2004

Plexins are cell surface receptors for semaphorin molecules, and their interaction governs cell adhesion and migration in a variety of tissues. We report that the Semaphorin 4D (Sema4D) receptor Plexin-B1 directly stimulates the intrinsic guanosine triphosphatase (GTPase) activity of R-Ras, a member of the Ras superfamily of small GTP-binding proteins that has been implicated in promoting cell adhesion and neurite outgrowth. This activity required the interaction of Plexin-B1 with Rnd1, a small GTP-binding protein of the Rho family. Down-regulation of R-Ras activity by the Plexin-B1-Rnd1 complex was essential for the Sema4D-induced growth cone collapse in hippocampal neurons. Thus, Plexin-B1 mediates Sema4D-induced repulsive axon guidance signaling by acting as a GTPase activating protein for R-Ras.
Multiplex Detection of RNA Expression in
Drosophila
Embryos
Authors: Kosman Dave , Mizutani Claudia M. , Lemons Derek , Cox W. Gregory , McGinnis William , Bier Ethan
Published: August 2004

We present a fluorescence-based, multiplex in situ hybridization method that permits the simultaneous detection of five differently labeled antisense RNA probes and up to seven different transcripts in a single
Drosophila
embryo. We also show that it should be possible to increase the number of detected transcripts substantially with nascent transcript multiplex fluorescent in situ hybridization. These multiplex methods fill a current technological gap between high-resolution in situ hybridization with one or two fluorescently labeled probes and low-resolution but genome-wide microarray RNA profiling and should be of great utility in establishing gene networks.
Surficial Deposits at Gusev Crater Along Spirit Rover Traverses
Authors: Grant J. A. , Arvidson R. , Bell J. F. , Cabrol N. A. , Carr M. H. , Christensen P. , Crumpler L. , Des Marais D. J. , Ehlmann B. L. , Farmer J. , Golombek M. , Grant F. D. , Greeley R. , Herkenhoff K. , Li R. , McSween H. Y. , Ming D. W. , Moersch J. , Rice J. W. , Ruff S. , Richter L. , Squyres S. , Sullivan R. , Weitz C.
Published: August 2004

The Mars Exploration Rover Spirit has traversed a fairly flat, rock-strewn terrain whose surface is shaped primarily by impact events, although some of the landscape has been altered by eolian processes. Impacts ejected basaltic rocks that probably were part of locally formed lava flows from at least 10 meters depth. Some rocks have been textured and/or partially buried by windblown sediments less than 2 millimeters in diameter that concentrate within shallow, partially filled, circular impact depressions referred to as hollows. The terrain traversed during the 90-sol (martian solar day) nominal mission shows no evidence for an ancient lake in Gusev crater.
Multiple Rare Alleles Contribute to Low Plasma Levels of HDL Cholesterol
Authors: Cohen Jonathan C. , Kiss Robert S. , Pertsemlidis Alexander , Marcel Yves L. , McPherson Ruth , Hobbs Helen H.
Published: August 2004

Heritable variation in complex traits is generally considered to be conferred by common DNA sequence polymorphisms. We tested whether rare DNA sequence variants collectively contribute to variation in plasma levels of highdensity lipoprotein cholesterol (HDL-C). We sequenced three candidate genes (
ABCA1, APOA1
, and
LCAT
) that cause Mendelian forms of low HDL-C levels in individuals from a population-based study. Nonsynonymous sequence variants were significantly more common (16% versus 2%) in individuals with low HDL-C (95th percentile). Similar findings were obtained in an independent population, and biochemical studies indicated that most sequence variants in the low HDL-C group were functionally important. Thus, rare alleles with major phenotypic effects contribute significantly to low plasma HDL-C levels in the general population.
Pancam Multispectral Imaging Results from the Spirit Rover at Gusev Crater
Authors: Bell J. F. , Squyres S. W. , Arvidson R. E. , Arneson H. M. , Bass D. , Blaney D. , Cabrol N. , Calvin W. , Farmer J. , Farrand W. H. , Goetz W. , Golombek M. , Grant J. A. , Greeley R. , Guinness E. , Hayes A. G. , Hubbard M. Y. H. , Herkenhoff K. E. , Johnson M. J. , Johnson J. R. , Joseph J. , Kinch K. M. , Lemmon M. T. , Li R. , Madsen M. B. , Maki J. N. , Malin M. , McCartney E. , McLennan S. , McSween H. Y. , Ming D. W. , Moersch J. E. , Morris R. V. , Dobrea E. Z. Noe , Parker T. J. , Proton J. , Rice J. W. , Seelos F. , Soderblom J. , Soderblom L. A. , Sohl-Dickstein J. N. , Sullivan R. J. , Wolff M. J. , Wang A.
Published: August 2004

Panoramic Camera images at Gusev crater reveal a rock-strewn surface interspersed with high- to moderate-albedo fine-grained deposits occurring in part as drifts or in small circular swales or hollows. Optically thick coatings of fine-grained ferric ironârich dust dominate most bright soil and rock surfaces. Spectra of some darker rock surfaces and rock regions exposed by brushing or grinding show near-infrared spectral signatures consistent with the presence of mafic silicates such as pyroxene or olivine. Atmospheric observations show a steady decline in dust opacity during the mission, and astronomical observations captured solar transits by the martian moons, Phobos and Deimos, as well as a view of Earth from the martian surface.
Initial Results from the Mini-TES Experiment in Gusev Crater from the Spirit Rover
Authors: Christensen P. R. , Ruff S. W. , Fergason R. L. , Knudson A. T. , Anwar S. , Arvidson R. E. , Bandfield J. L. , Blaney D. L. , Budney C. , Calvin W. M. , Glotch T. D. , Golombek M. P. , Gorelick N. , Graff T. G. , Hamilton V. E. , Hayes A. , Johnson J. R. , McSween H. Y. , Mehall G. L. , Mehall L. K. , Moersch J. E. , Morris R. V. , Rogers A. D. , Smith M. D. , Squyres S. W. , Wolff M. J. , Wyatt M. B.
Published: August 2004

The Miniature Thermal Emission Spectrometer (Mini-TES) on Spirit has studied the mineralogy and thermophysical properties at Gusev crater. Undisturbed soil spectra show evidence for minor carbonates and bound water. Rocks are olivinerich basalts with varying degrees of dust and other coatings. Dark-toned soils observed on disturbed surfaces may be derived from rocks and have derived mineralogy (Â±5 to 10%) of 45% pyroxene (20% Ca-rich pyroxene and 25% pigeonite), 40% sodic to intermediate plagioclase, and 15% olivine (forsterite 45% Â±5 to 10). Two spectrally distinct coatings are observed on rocks, a possible indicator of the interaction of water, rock, and airfall dust. Diurnal temperature data indicate particle sizes from 40 to 80 Î¼m in hollows to â¼0.5 to 3 mm in soils.
Clinical and Epidemiological Aspects of Infections Caused byFusariumSpecies: a Collaborative Study from Israel
Authors: Nir-Paz Ran , Strahilevitz Jacob , Shapiro Mervyn , Keller Nathan , Goldschmied-Reouven Anna , Yarden Oded , Block Colin , Polacheck Itzhack
Published: August 2004

Fusariuminfections are an important problem worldwide, commonly affecting immunocompromised individuals. We conducted a retrospective study in two Israeli tertiary medical centers of factors predisposing to infection byFusariumspp. and their influence on the epidemiology and clinical outcome of this infection.Fusariumspp. were isolated from 89 patients with a median age of 57 years. Sixty-eight patients were considered immunocompetent. Seven patients had disseminated disease, 34 had locally invasive disease, and 48 had superficial infection. Most infections were limited and occurred mainly in lower limbs. Factors associated with in-hospital mortality were chronic renal failure, hematological malignancy, immunosuppression, disseminated infection, and positive blood culture. Multivariate analysis showed that chronic renal failure, hematological malignancy, burns, and disseminated infection were independently associated with mortality. A surge in the frequency of infections was noticed during the summer for patients from rural areas, involving mainly the eyes and lungs. At one of the hospitals (in a mountainous area), there was an increase in the isolation rate over time.
Bartonella koehlerae
, a New Cat-Associated Agent of Culture-Negative Human Endocarditis
Authors: Avidor Boaz , Graidy Merav , Efrat Gabi , Leibowitz Cecilia , Shapira Gregory , Schattner Ami , Zimhony Oren , Giladi Michael
Published: August 2004

Bartonella koehlerae
is reported for the first time to be a human pathogen that causes culture-negative endocarditis. It is also shown that this species, isolated twice before from domestic cats, can be recovered as well from a stray cat population in Israel. This work follows a recent report of the same case in which the causative agent was misidentified as
B. henselae
, based on serology and PCR-restriction fragment length polymorphism (RFLP) analysis (A. Schattner, O. Zimhony, B. Avidor, and M. Gilad, Lancet 361:1786, 2003).
B. koehlerae
was identified in the valvular tissue of an endocarditis patient by DNA sequencing of the PCR products of two
Bartonella
genes: the genes for citrate synthase (
gltA
) and riboflavin synthase (
ribC
). The commonly used PCR-RFLP analysis of the TaqI-digested
gltA
PCR product did not distinguish between
B. koehlerae
and
B. quintana
or between
B. elizabethae
and
B. clarridgeiae
. PmlI digestion of the
gltA
amplification product failed to differentiate between
B. quintana
,
B. clarridgeiae
, and
B. elizabethae
. RFLP analysis of the heat shock protein (
htrA
) gene by TaqI digestion misidentified
B. koehlerae
as
B. henselae
. However, RFLP analysis of the
ribC
PCR product, digested with TaqI, was able to distinguish between the human endocarditis-associated
Bartonella
species tested,
B. henselae
,
B. quintana
,
B. elizabethae
, and
B. koehlerae
, as well as between the cat-associated
Bartonella
species,
B. henselae
and
B. clarridgeiae
. Given the expanding number of
Bartonella
species emerging as human pathogens, it is suggested that PCR-RFLP analysis for the diagnosis of
Bartonella
infections target several genes and be coupled with DNA sequencing to avoid species identification.
Clinical Evaluation of the Digene Hybrid Capture II Test and the COBAS AMPLICOR Monitor Test for Determination of Hepatitis B Virus DNA Levels
Authors: Yuan He-Jun , Yuen Man-Fung , Wong Danny Ka-Ho , Sum Simon Siu-Man , Lai Ching-Lung
Published: August 2004

The measurement of hepatitis B virus (HBV) DNA is important for the assessment of liver disease and treatment efficacy. Most commercially available assays for the determination of HBV DNA levels have limited linear ranges. This study was performed to evaluate the clinical performance of the Digene Hybrid Capture II (Digene HC II assay) and the COBAS AMPLICOR Monitor test (COBAS-AM assay), with special emphasis on anti-HBV e antigen (HBeAg)-positive patients with low HBV DNA levels. A total of 425 Chinese patients with chronic hepatitis B were recruited. A total of 107 patients were HBeAg positive, and 318 patients were HBeAg negative. The Digene HC II assay and the COBAS-AM assay had similar intra-assay and interassay variabilities. A total of 264 patients (62.1%) had HBV DNA levels undetectable by the Digene HC II assay, and 47 patients (11.1%) had HBV DNA levels undetectable by the COBAS-AM assay (
P
< 0.001). For the 161 patients with HBV DNA levels detectable by the Digene HC II assay, the HBV DNA levels obtained by the Digene HC II assay and by the COBAS-AM assay showed an excellent correlation (
r
= 0.95;
P
< 0.001). The linear ranges of the Digene HC II assay and the COBAS-AM assay marginally overlapped. Before HBV DNA levels could be determined by the COBAS-AM assay, predilution had to be performed for 158 of 161 patients (98.1%) with HBV DNA levels detectable by the Digene HC II assay and for 10 of 264 patients (3.8%) with HBV DNA levels undetectable by the Digene HC II assay. The cost for assaying each serum sample by using different strategies was calculated. The COBAS-AM assay was more sensitive than the Digene HC II assay and more suitable for monitoring low levels of HBV viremia.
Comparison of the BBL CHROMagar Staph aureus Agar Medium to Conventional Media for Detection of
Staphylococcus aureus
in Respiratory Samples
Authors: Flayhart Diane , Lema Clara , Borek Anita , Carroll Karen C.
Published: August 2004

Screening for
Staphylococcus aureus
has become routine in certain patient populations. This study is the first clinical evaluation of the BBL CHROMagar Staph aureus agar (CSA) medium (BD Diagnostics, Sparks, Md.) for detection of
S. aureus
in nasal surveillance cultures and in respiratory samples from cystic fibrosis (CF) patients.
S. aureus
colonies appear mauve on CSA. Other organisms are inhibited or produce a distinctly different colony color.
S. aureus
was identified from all media by slide coagulase, exogenous DNase, and mannitol fermentation assays. Susceptibility testing was performed using the agar dilution method. A total of 679 samples were evaluated. All samples were inoculated onto CSA. Nasal surveillance cultures were inoculated onto sheep blood agar (SBA) (BD Diagnostics), and samples from CF patients were inoculated onto mannitol salt agar (MSA) (BD Diagnostics). Of the 679 samples cultured, 200 organisms produced a mauve color on CSA (suspicious for
S. aureus
) and 180 were positive for
S. aureus
on SBA or MSA. Of 200 CSA-positive samples 191 were identified as
S. aureus
. Nine mauve colonies were slide coagulase negative and were subsequently identified as
Staphylococcus lugdunensis
(one),
Staphylococcus epidermidis
(three),
Staphylococcus haemolyticus
(one), and
Corynebacterium
species (four). CSA improved the ability to detect
S. aureus
by recovering 12
S. aureus
isolates missed by conventional media. Of the 192
S. aureus
isolates recovered, 122 were methicillin susceptible and 70 were methicillin resistant. Overall, the sensitivity and specificity of CSA in this study were 99.5 and 98%, respectively. There was no difference in the performance of the slide coagulase test or in susceptibility testing performed on
S. aureus
recovered from CSA compared to SBA or MSA. Our data support the use of CSA in place of standard culture media for detection of
S. aureus
in heavily contaminated respiratory samples.
Use of Fluorescent Amplified Fragment Length Polymorphism for Molecular Epidemiology of Leptospirosis in India
Authors: Vijayachari P. , Ahmed Niyaz , Sugunan A. P. , Ghousunnissa Sheikh , Rao K. Rajender , Hasnain Seyed E. , Sehgal Subhash C.
Published: August 2004

Nineteen isolates of leptospires recovered from patients during three epidemics that occurred at different places and different times in the Andaman Islands and eight isolates from sporadic cases were characterized using serological and molecular genetic techniques. Group sera and monoclonal antibodies were used for antigenic characterization, whereas fluorescent amplified fragment length polymorphism (FAFLP) was used for genotyping. Of the 27 isolates, 19 were identified as belonging to serogroup Grippotyphosa, 3 belonged to serogroup Australis, 2 belonged to serogroup Icterohaemorrhagiae, and 1 each belonged to serogroups Hebdomadis, Canicola, and Sejroe. Analysis of FAFLP data grouped these 27 isolates into two main clusters of genotypes. One of the clusters, populated by 19 isolates, included 16 outbreak isolates. Seven of these 19 isolates belonged to serovar Ratnapura, 10 belonged to serovar Valbuzzi, and 1 each belonged to serovar Grippotyphosa and serovar Saxkoebing. Of the 27 patients from whom isolates were obtained, 9 had severe illness, and 6 of these 9 patients had pulmonary involvement, 1 had pulmonary and hepatorenal involvement, and the remaining 2 had hepatorenal involvement alone. Two patients out of the nine severe cases died subsequently. The isolates from sporadic cases showed great genetic diversity and were also diverse antigenically. Perhaps the strains belonging to a dominant genotype (the outbreak-associated cluster) possessed epidemic potential and higher virulence with a greater predilection to cause pulmonary complications than strains belonging to other genetic backgrounds.
Molecular Epidemiology of Hospital-Associated and Community-Acquired
Clostridium difficile
Infection in a Swedish County
Authors: Norén T. , Åkerlund T. , Bäck E. , Sjöberg L. , Persson I. , Alriksson I. , Burman L. G.
Published: August 2004

All episodes of
Clostridium difficile
associated diarrhea (CDAD) diagnosed in a defined population of 274,000 including one tertiary and two primary hospitals and their catchment areas were studied during 12 months. The annual CDAD incidence in the county was 97 primary episodes per 100,000, and 78% of all episodes were classified as hospital associated with a mean incidence of 5.3 (range, 1.4 to 6.5) primary episodes per 1,000 admissions. The incidence among hospitalized individuals was 1,300-fold higher than that in the community (33,700 versus 25 primary episodes per 100,000 persons per year), reflecting a 37-fold difference in antibiotic consumption (477 versus 13 defined daily doses [DDD]/1,000 persons/day) and other risk factors. Three tertiary hospital wards with the highest incidence (13 to 36 per 1,000) had CDAD patients of high age (median age of 80 years versus 70 years for other wards,
P
< 0.001), long hospital stay (up to 25 days versus 4 days), or a high antibiotic consumption rate (up to 2,427 versus 421 DDD/1,000 bed days). PCR ribotyping of
C. difficile
isolates available from 330 of 372 CDAD episodes indicated nosocomial acquisition of the strain in 17 to 27% of hospital-associated cases, depending on the time interval between index and secondary cases allowed (2 months or up to 12 months), and only 10% of recurrences were due to a new strain of
C. difficile
(apparent reinfection). In other words, most primary and recurring episodes were apparently caused by the patient's endogenous strain rather than by one of hospital origin. Typing also indicated that a majority of
C. difficile
strains belonged to international serotypes, and the distribution of types was similar within and outside hospitals and in primary and relapsing CDAD. However, type SE17 was an exception, comprising 22% of hospital isolates compared to 6% of community isolates (
P
= 0.008) and causing many minor clusters and a silent nosocomial outbreak including 36 to 44% of the CDAD episodes in the three high-incidence wards.
New Genome Type of Adenovirus Serotype 4 Caused Nosocomial Infections Associated with Epidemic Conjunctivitis in Japan
Authors: Ariga Toshihide , Shimada Yasushi , Ohgami Kazuhiro , Tagawa Yoshitsugu , Ishiko Hiroaki , Aoki Koki , Ohno Shigeaki
Published: August 2004

Human adenovirus type 4 is one of the major serotypes isolated from patients with adenoviral conjunctivitis. In 2001 we encountered nosocomial infections with epidemic conjunctivitis in the ophthalmology ward of one hospital in Sapporo, which is in the northern part of Japan. Adenoviruses were isolated from the patients with this nosocomial infection and identified as adenovirus type 4 (AdV-4) by a neutralization test with serotype-specific antiserum. When the cleavage patterns of the isolates were compared with the full viral genome with BamHI and SmaI, the cleavage patterns of the isolates were shown to be different from those of AdV-4p and other previously known AdV-4 variants. The nucleotide sequences of the fiber gene of the isolates showed the highest homologies (94.3%) with AdV-4 among the nucleotide sequences available from GenBank and formed a monophyletic cluster along with the prototype strain of AdV-4. The isolates, however, were located in a different lineage from those of AdV-4p and the AdV-4 variant from the sporadic infections. We conclude that the nosocomial infection that appeared in 2001 was caused by a new genome type of AdV-4, which was designated AdV-4c.
Comparison of a New Lateral-Flow Chromatographic Membrane Immunoassay to Viral Culture for Rapid Detection and Differentiation of Influenza A and B Viruses in Respiratory Specimens
Authors: Cazacu Andreea C. , Demmler Gail J. , Neuman Mark A. , Forbes Betty A. , Chung Sooyoung , Greer Jewel , Alvarez Ana E. , Williams Robin , Bartholoma Nadine Y.
Published: August 2004

The performance of a new rapid lateral-flow chromatographic membrane immunoassay test kit for detection of influenza virus was evaluated and compared to that of viral culture in respiratory secretions collected from 400 adults and children seen at three large university hospitals during the recent 2003 influenza season. The rapid test provided results in 15 min, with excellent overall performance statistics (sensitivity, 94.4%; specificity, 100%; positive predictive value, 100%; negative predictive value, 97.5%). Both influenza A and B type viruses were reliably detected, with no significant difference in performance statistics noted by influenza virus type or by the center performing the test.
Recommended Test Panel for Differentiation of
Klebsiella
Species on the Basis of a Trilateral Interlaboratory Evaluation of 18 Biochemical Tests
Authors: Hansen Dennis S. , Aucken Hazel M. , Abiola Titi , Podschun Rainer
Published: August 2004

Klebsiella pneumoniae
and
Klebsiella oxytoca
are the two most frequently encountered
Klebsiella
species giving rise to infections in humans, but other
Klebsiella
species can also be found in clinical specimens:
Klebsiella ozaenae
,
Klebsiella rhinoscleromatis
,
Klebsiella terrigena
,
Klebsiella planticola
,
Klebsiella ornithinolytica
, and
Enterobacter aerogenes
(
Klebsiella mobilis
). However, many of these species are indistinguishable by the conventional methods employed routinely in the clinical microbiological laboratory. Several investigators have suggested various additional tests, but as yet there is no standardized test panel for identifying all
Klebsiella
species and subspecies. In the present study, performed in three national
Klebsiella
reference laboratories, we have evaluated a test panel consisting of 18 biochemical tests on 242 strains comprising all
Klebsiella
species and subspecies. The test panel was designed to identify organisms preliminarily identified as belonging to the genus
Klebsiella
on the basis of conventional methods or automated identification systems. With the described test panel it is possible to find one or more positive test results differentiating any
Klebsiella
species, except
Klebsiella rhinoscleromatis
, from its closest relative.
Comparison of Oligonucleotide Ligation Assay and Consensus Sequencing for Detection of Drug-Resistant Mutants of Human Immunodeficiency Virus Type 1 in Peripheral Blood Mononuclear Cells and Plasma
Authors: Ellis Giovanina M. , Mahalanabis Madhumita , Beck Ingrid A. , Pepper Gregory , Wright Amy , Hamilton Shannon , Holte Sarah , Naugler Willscott E. , Pawluk Diane M. , Li Chung-Chen , Frenkel Lisa M.
Published: August 2004

Drug-resistant mutants of human immunodeficiency virus type 1 (HIV-1) recede below the limit of detection of most assays applied to plasma when selective pressure is altered due to changes in antiretroviral treatment (ART). Viral variants with different mutations are selected by the new ART when replication is not suppressed or wild-type variants with greater replication fitness outgrow mutants following the cessation of ART. Mutants selected by past ART appear to persist in reservoirs even when not detected in the plasma, and when conferring cross-resistance they can compromise the efficacy of novel ART. Oligonucleotide ligation assay (OLA) of virus in plasma and peripheral blood mononuclear cells (PBMC) was compared to consensus sequence dideoxynucleotide chain terminator sequencing for detection of 91 drug resistance mutations that had receded below the limit of detection by sequencing of plasma. OLA of plasma virus detected 27.5% (95% confidence interval [CI], 19 to 39%) of mutant genotypes; consensus sequencing of the PBMC amplicon from the same specimen detected 23.1% (95% CI, 14 to 34%); and OLA of PBMC detected 53.8% (95% CI, 44 to 64%). These data suggest that concentrations of drug-resistant mutants were greater in PBMC than in plasma after changes in ART and indicate that the OLA was more sensitive than consensus sequencing in detecting low levels of select drug-resistant mutants.
Use of 16S rRNA, 23S rRNA, and
gyrB
Gene Sequence Analysis To Determine Phylogenetic Relationships of
Bacillus cereus
Group Microorganisms
Authors: Bavykin Sergei G. , Lysov Yuri P. , Zakhariev Vladimir , Kelly John J. , Jackman Joany , Stahl David A. , Cherni Alexey
Published: August 2004

In order to determine if variations in rRNA sequence could be used for discrimination of the members of the
Bacillus cereus
group, we analyzed 183 16S rRNA and 74 23S rRNA sequences for all species in the
B. cereus
group. We also analyzed 30
gyrB
sequences for
B. cereus
group strains with published 16S rRNA sequences. Our findings indicated that the three most common species of the
B. cereus
group,
B. cereus, Bacillus thuringiensis
, and
Bacillus mycoides
, were each heterogeneous in all three gene sequences, while all analyzed strains of
Bacillus anthracis
were found to be homogeneous. Based on analysis of 16S and 23S rRNA sequence variations, the microorganisms within the
B. cereus
group were divided into seven subgroups, Anthracis, Cereus A and B, Thuringiensis A and B, and Mycoides A and B, and these seven subgroups were further organized into two distinct clusters. This classification of the
B. cereus
group conflicts with current taxonomic groupings, which are based on phenotypic traits. The presence of
B. cereus
strains in six of the seven subgroups and the presence of
B. thuringiensis
strains in three of the subgroups do not support the proposed unification of
B. cereus
and
B. thuringiensis
into one species. Analysis of the available phenotypic data for the strains included in this study revealed phenotypic traits that may be characteristic of several of the subgroups. Finally, our results demonstrated that rRNA and
gyrB
sequences may be used for discriminating
B. anthracis
from other microorganisms in the
B. cereus
group.
Performance Characteristics of a Quantitative TaqMan Hepatitis C Virus RNA Analyte-Specific Reagent
Authors: Barbeau James M. , Goforth Jennifer , Caliendo Angela M. , Nolte Frederick S.
Published: August 2004

We determined the dynamic range, reproducibility, accuracy, genotype bias, and sensitivity of the TaqMan hepatitis C virus (HCV) analyte-specific reagent (ASR). Serum samples were processed using the MagNA Pure LC instrument and run on the COBAS TaqMan 48 analyzer. The performance characteristics of the ASR were also compared with those of the qualitative AMPLICOR and quantitative AMPLICOR MONITOR HCV tests. The ASR exhibited a ≥6-log
10
linear dynamic range and excellent reproducibility, with a mean coefficient of variation of 14%. HCV RNA concentration measured with the ASR agreed within an average of 0.42 log
10
(2.6-fold) of the labeled concentration with members of a standard reference panel. HCV genotypes 1 to 4 were amplified with similar efficiencies with the ASR. The ASR and AMPLICOR MONITOR viral load results were significantly correlated (
r
= 0.8898;
P
< 0.01), but the agreement was poor (mean difference, 0.45 ± 0.35 log
10
) for 72 HCV RNA-positive clinical samples. However, 98.9% agreement between the ASR and qualitative AMPLICOR test results was found with 60 positive and 29 negative samples. Limiting-dilution experiments demonstrated that the limits of detection for ASR and AMPLICOR tests were 84 and 26 IU/ml, respectively. The performance characteristics of the TaqMan HCV ASR are appropriate for all clinical applications of HCV RNA testing.
Multicenter Evaluation of Use of Penicillin and Ampicillin as Surrogates for In Vitro Testing of Susceptibility of Enterococci to Imipenem
Authors: Weinstein Melvin P. , Mirrett Stanley , Kannangara Saman , Monahan Jan , Harrell Lizzie J. , Wilson Alan C. , Reller L. Barth
Published: August 2004

Imipenem is approved by the U.S. Food and Drug Administration (FDA) for treatment of infections caused by
Enterococcus faecalis
. However, there are no NCCLS guidelines for testing susceptibility of enterococci against imipenem. To assess whether or not ampicillin or penicillin could be used as a surrogate for broth microdilution (BMD) testing of imipenem versus
Enterococcus
species, 633 strains of
E. faecalis
,
E. faecium
, and other enterococci isolated from blood cultures of patients at three geographically distinct university hospitals were tested by the NCCLS BMD and disk diffusion (DD) methods. Using FDA susceptibility breakpoints for imipenem and NCCLS breakpoints for penicillin and ampicillin, categorical agreement (CA) for penicillin-imipenem and ampicillin-imipenem tested with
E. faecalis
and
E. faecium
by BMD was ≥94% but was ≤90% for other enterococci. Using the DD method, CA for ampicillin-imipenem tested with
E. faecalis
and
E. faecium
was ≥98% and was 92% for other enterococci; CA for penicillin-imipenem was 91% for
E. faecalis
, 98% for
E. faecium
, and 87% for other enterococci. Further analysis showed that testing
E. faecalis
with ampicillin resulted in no false-susceptible (FS) or false-resistant (FR) results by BMD, no FS results by DD, and a single FR result by DD (0.2%), whereas testing with penicillin resulted in no FS results by BMD or DD and two FR results by BMD (0.4%). For
E. faecium
and other enterococci, the combination of FS and FR results was such that surrogate testing with penicillin or ampicillin appears not to be sufficiently reliable to be used clinically. We conclude that ampicillin is an accurate predictor of the in vitro activity of imipenem against
E. faecalis
.
Molecular Characteristics of Nosocomial and Native American Community-Associated Methicillin-ResistantStaphylococcus aureusClones from Rural Wisconsin
Authors: Shukla Sanjay K. , Stemper Mary E. , Ramaswamy Srinivas V. , Conradt Jennifer M. , Reich Robert , Graviss Edward A. , Reed Kurt D.
Published: August 2004

In central and northern Wisconsin methicillin-resistantStaphylococcus aureus(MRSA) was first detected in 1989. Over the next 10-year period, 581 MRSA isolates were collected, 17.2% of which came from patients who were treated at five Native American clinics. These isolates were typed by SmaI-macrorestricted pulsed-field gel electrophoresis (PFGE). The PFGE patterns clustered the isolates into six major clonal groups (MCGs), i.e., MCGs 1 to 6, and 19 minor clonal groups (mCGs). The 25 clonal groups were represented by 109 unique PFGE types. Sixty-five percent of the MCG-2 isolates were recovered from patients who were treated at Native American clinics. Ninety-four percent of the MCG-2 isolates harbored the staphylococcal cassette chromosomemec(SCCmec) IVa. These isolates also had PFGE profiles that were clonally related to the midwestern community-associated MRSA (CA-MRSA) strain, MW2. The representative isolates from MCG-2 had the multilocus sequence type allelic profile 1-1-1-1-1-1-1 and containedpvlgenes. They were also susceptible to various antibiotics, a finding consistent with the CA-MRSA phenotype. SCCmecIV was also present in othermCGs. Unlike MCG-2, isolates from the remaining five MCGs harbored SCCmecII and were resistant to multiple antibiotics, suggesting their nosocomial origin. The 19mCGs were represented by diverse SCCmectypes and three putative new variants referred to as SCCmecIb, IIa, and IIb.
Detection of
Haemophilus influenzae
Type b by Real-Time PCR
Authors: Marty Adrian , Greiner Oliver , Day Philip J. R. , Gunziger Sibylle , Mühlemann Kathrin , Nadal David
Published: August 2004

A real-time PCR assay targeting the capsulation locus of
Haemophilus influenzae
type b (Hib) was developed. The linear detection range was from 1 to 10
6
microorganisms per reaction mixture. No
H. influenzae
other than Hib or any other control bacteria typically found in the upper respiratory tract was detected.
Isolation of
Bartonella rattimassiliensis
sp. nov. and
Bartonella phoceensis
sp. nov. from European
Rattus norvegicus
Authors: Gundi Vijay A. K. B. , Davoust Bernard , Khamis Atieh , Boni Mickaël , Raoult Didier , La Scola Bernard
Published: August 2004

Thirty-three isolates of
Bartonella
spp., including 11 isolates not belonging to previously known species, were isolated from 66
Rattus norvegicus
subjects trapped in the city of Marseille, France. Based on seven different gene sequences, the 11 isolates were assigned to
Bartonella rattimassiliensis
sp. nov. and
Bartonella phoceensis
sp. nov.
Analysis of
p51
,
groESL
, and the Major Antigen P51 in Various Species of
Neorickettsia
, an Obligatory Intracellular Bacterium That Infects Trematodes and Mammals
Authors: Rikihisa Yasuko , Zhang Chunbin , Kanter Manuel , Cheng Zhihui , Ohashi Norio , Fukuda Takeo
Published: August 2004

The
p51
gene that encodes the major antigenic 51-kDa protein in
Neorickettsia risticii
was identified in strains of
Neorickettsia sennetsu
and the
Stellantchasmus falcatus
agent but not in
Neorickettsia helminthoeca
, suggesting that
p51
-based diagnosis would be useful to distinguish among them.
groESL
sequencing results delineated the phylogenic relationships among
Neorickettsia
spp.
Real-Time TaqMan PCR for Quantifying Oral Bacteria during Biofilm Formation
Authors: Suzuki Nao , Nakano Yoshio , Yoshida Akihiro , Yamashita Yoshihisa , Kiyoura Yusuke
Published: August 2004

A TaqMan PCR was developed for quantifying early colonizer microorganisms in dental biofilms. To design species-specific primers and TaqMan probes, genomic subtractive hybridization was used. This quantitative assay in combination with subtractive hybridization may be of value in the study of microbial ecosystems consisting of related species that are involved in the formation and etiology of biofilms.