bluuebunny/crossref_metadata_2025 huggingface dataset:
Pink bollworm moth (Lepidoptera: Gelechiidae) catches in the Imperial Valley, California from 1989 to 2003
Authors: CHU CHANG‐CHI , NATWICK ERIC T. , LÓPEZ RAUL LEÓN , DESSERT JOLENE R. , HENNEBERRY THOMAS J.
Published: December 2006

Abstract
We examined the patterns of male pink bollworm (PBW), Pectinophora gossypiella (Saunders), moth catches in gossyplure‐baited traps over a 15‐year period from 1989 to 2003 in the Imperial Valley, California, USA. Monitoring was conducted during periods when different pink bollworm areawide control strategies were being used. Numbers of male pink bollworm moths caught in gossyplure‐baited traps progressively decreased each year from 1990 to 1994 during short‐season cotton production. High numbers of male moths caught in traps from 1995 to 1997 may have been related to moth migrations from the large cotton acreages grown in the Mexicali Valley bordering the Imperial Valley. Transgenic Bollgard® (Bt) cotton was planted in 3% of the cotton area in 1996 and thereafter in 80%–94% of the cotton area from 1997 to 2003. Pink bollworm moth trap catches were significantly lower from 1998 to 2003 than catches in 1995 to 1997, except for 1999. The trapping results suggested that Bt cotton had significant input on reduction of pink bollworm populations, confirming results of other investigators and providing additional documentation on the benefits of the Bt cotton culture.
Comparative demography of Liriomyza sativae Blanchard (Diptera: Agromyzidae) on cucumber at seven constant temperatures
Authors: HAGHANI MOSTAFA , FATHIPOUR YAGHOUB , TALEBI ALI ASGHAR , BANIAMERI VALIOLLAH
Published: December 2006

Abstract
Reproduction and population parameters of vegetable leafminer, Liriomyza sativae Blanchard were measured on cucumber (Cucumis sativus L.) at seven constant temperatures (10, 15, 20, 25, 30, 35 and 40°C). No eggs were found at 10°C and flies died after exposure to 40°C. The significantly highest intrinsic rate of natural increase (rm), net reproductive rate (R0) and finite rate of increase (δ) of L. sativae were obtained at 25°C as 0.196, 52.452, and 1.216, respectively. The above‐mentioned parameters decreased at 15°C and 35°C and this reduction at 35°C was strong. Doubling time (DT) varied significantly with temperature. The shortest doubling time was obtained at 25°C. Mean generation time (T) decreased significantly with increasing temperature between 15°C and 35°C. Percentage of immature ages in the stable age distribution was more than 95% at all temperatures. Female longevity was greater than male at all temperatures. Liriomyza sativae lived for a long time at 15°C, whereas at 35°C had lower survival rates. The effect of temperature on reproduction, especially the intrinsic rate of increase of L. sativae would be useful for predicting its long‐term population fluctuation over several generations.
Feeding and oviposition preference of Helopeltis theivora (Hemiptera: Miridae) on tea in Northeast India
Authors: BHUYAN MANTU , BHATTACHARYYA PRANAB R.
Published: December 2006

Abstract
Feeding and oviposition preference studies of Helopeltis theivora Waterhouse, on leaves and stems of tea, Camellia sinensis L. (O) Kuntze revealed that the insect mostly preferred the second leaf for feeding in comparison to the first, and the third leaf and stems; while the fifth instar nymphs were the most voracious feeders among the life stages, producing the most and largest feeding lesions. The insect preferred to oviposit mainly on stems and very few eggs are found on leaves.
Synthesis andIn VitroEvaluation of Novel 2‐oxo‐1,2‐dihydroquinoline CB2 Receptor Inverse Agonists
Authors: Raitio Katri H. , Savinainen Juha R. , Nevalainen Tapio , Järvinen Tomi , Vepsäläinen Jouko
Published: December 2006

Aliphatic amides of 7‐methoxy‐2‐oxo‐8‐pentyloxy‐1,2‐dihydroquinoline‐3‐carboxylic acid 6 were synthesized and evaluated for their CB2 and CB1 receptor activities by a [35S]GTPS membrane‐binding assay. Compounds 7–18 produced CB2 inverse agonist activities with a variety of potencies (IC50: 1 nm–1 μm). By contrast, no considerable antagonism of CB1 receptor was observed by 1–10 μmconcentrations indicating that synthesized compounds were CB2 receptor selective.
Radiolabeling and in vitro and in vivo Characterization of [18F]FB‐[R8,15,21, L17]‐VIP as a PET Imaging Agent for Tumor Overexpressed VIP Receptors
Authors: Cheng Dengfeng , Yin Duanzhi , Li Gucai , Wang Mingwei , Li Shiqiang , Zheng Mingqiang , Cai Hancheng , Wang Yongxian
Published: December 2006

In an effort to develop a peptide‐based radiopharmaceutical for the detection of tumors overexpressed vasoactive intestinal peptide receptors with positron emission tomography, we have prepared a novel [R8,15,21, L17]‐VIP peptide for 18F‐labeling. This peptide inhibited 125I‐VIP binding to rats lung membranes with high affinity [half‐maximal inhibitory concentrations (IC50) of 0.12 nm]. Additionally, [R8,15,21, L17]‐VIP showed higher stability than native vasoactive intestinal peptide in vivo of mice. With N‐succinimidyl 4‐[18F] fluorobenzoate as labeling prosthetic group, [18F]FB‐[R8,15,21, L17]‐VIP was obtained in >99% radiochemical purity within 100 min in decay‐for‐corrected radiochemical yield of 33.6 ± 3% (n = 5) and a specific radioactivity 255 GBq/μmol at the end of synthesis. Stability of [18F]FB‐[R8,15,21, L17]‐VIP in vitro and in vivo were investigated. Biodistribution of this trace was carried out in mice with induced C26 colorectal tumor. Fast clearance of [18F]FB‐[R8,15,21, L17]‐VIP from non‐target tissues and specific uptakes by tumors realized higher tumor‐to‐muscle ratio (3.55) and tumor‐to‐blood ratio (2.37) 60 min postinjection. Clear difference was observed between the blocking and unblocking experiments in biodistribution and whole body radioautography. [18F]FB‐[R8,15,21, L17]‐VIP has demonstrated its potential for diagnosing tumors overexpressed vasoactive intestinal peptide receptors both in vitro and in vivo.
Influence of Generation 2–5 of PAMAM Dendrimer on the Inhibition of Tat Peptide/ TAR RNA Binding in HIV‐1 Transcription
Authors: Wang Wei , Guo Zhenpeng , Chen Yi , Liu Tao , Jiang Long
Published: December 2006

The special binding of Tat protein to TAR RNA leads to the transcription of HIV‐1 virus. In this study, the influence of 2–5 generation of PAMAM dendrimers on the inhibition of Tat protein/TAR RNA binding has been investigated. The absorption of PAMAM dendrimers on TAR RNA, fixed on a gold substrate through an avidin–biotion connection, was carried out by using a quartz crystal microbalance. Experimental result shows a Langmuir‐type isotherm could be used to describe this kind of binding, implying a specific and monolayer adsorption existed. The combination coefficient ()s can be calculated according to Langmuir Equation, having the order of G3 > G4 > G5 >Tat > G2, indicating that PAMAM G3, G4 and G5 having the possibility to be the inhibitors of HIV‐1 transcription. The migration time (Tmigra) of capillary electrophoretic technique has the same sequence as ()s. These two parameters could be used as simple and quantitative criteria for the selection of possible drugs from numerous candidates for HIV therapy in vitro.
4‐(Azolylphenyl)‐phthalazin‐1‐amines: Novel Inhibitors of VEGF Receptors I and II
Authors: Kiselyov Alexander S. , Semenov Victor V. , Milligan Daniel
Published: December 2006

Novel potent derivatives of phthalazine are described as ATP‐competitive inhibitors of vascular endothelial growth factor receptors I and II (VEGFR‐1/2). A number of compounds display VEGFR‐2 inhibitory activity reaching that of VatalanibTM3 (IC50 < 100 nm) in an HTRF enzymatic assay. Several derivatives also show good potential for the development as VEGFR‐2 specific inhibitors showing 15–20‐fold selectivity over VEGFR‐1.
Synthesis and Biological Activity of Seleno Sunflower Trypsin Inhibitor Analog
Authors: Guo Xin , Shi Jiahao , Tang Zhenyu , Cui Dafu , Zhang Youshang
Published: December 2006

Sunflower trypsin inhibitor (SFTI‐1) is a cyclic peptide with 14 amino acid residues and one disulfide bond. Its synthetic acyclic analog (aSFTI‐1) with N‐terminal Gly and C‐terminal Asp was still active. Here, we report the synthesis of seleno aSFTI‐1 with the disulfide bond of aSFTI‐1 replaced by diselenide bond. The formation of the diselenide bond from selenol was achieved in a single step without the aid of oxidizing agent. For comparison, aSFTI‐1 itself and aSFTI‐1 with its disulfide bond replaced by two serines ([Ser3,11] aSFTI‐1) were also synthesized. The trypsin inhibitory constants of seleno aSFTI‐1, aSFTI‐1 and [Ser3,11] aSFTI‐1 were determined as 6.50 × 10−9, 1.96 × 10−9 and 8.10 × 10−6 respectively, indicating that the disulfide bond is essential for the structure and function of aSFTI‐1, and seleno aSFTI‐1 is still active, although its inhibitory constant is reduced to 30% in comparison with that of aSFTI‐1.
Alternate Binding Mode of C‐terminal Phenethylamine Analogs of Gtα(340–350) to Photoactivated Rhodopsin
Authors: Anderson Matthew A. , Ogbay Benhur , Kisselev Oleg G. , Cistola David P. , Marshall Garland R.
Published: December 2006

The C‐terminus of the Gα‐subunit of transducin plays an important role in receptor recognition. Synthetic peptides corresponding to the last 11 residues of the subunit have been shown to stabilize the photoactivated form of rhodopsin, Rh*. The Rh*‐bound structure of the Gtα(340–350) peptide has been determined using transferred nuclear overhauser effect NMR. In that structure, we observed two interactions between Lys341 and Phe350, a cation–π interaction between the ɛ‐amine and the aromatic ring of Phe350 and a salt‐bridge between the ɛ‐amine and the C‐terminal carboxylate. A series of C‐terminal phenethylamine analogs of the Gtα(340–350) peptide were synthesized, lacking the C‐terminal carboxylate group, to investigate the forces that contribute to the stability of the Rh*‐bound conformation of the peptide. Rh*‐stabilization assay data suggest that the C‐terminal carboxylate is not necessary to maintain binding affinity. Transferred nuclear overhauser effect NMR experiments reveal that these C‐terminal phenethylamine peptides adopt an Rh*‐bound structure that is similar overall, but lacking some of the intramolecular interactions observed in the native Rh*‐bound Gtα(340–350) structure. These studies suggest that the binding site for Gtα(340–350) on Rh* is adaptable, and we propose that the charged carboxylate of Phe350 does not play a significant role in the interaction with Rh*, but helps stabilize the Rh*‐bound confirmation of the native peptide.
Environmental degradation problems in the Heihe River Basin, northwest China
Authors: Qi Shanzhong , Luo Fang
Published: December 2006

Environmental degradation in the Heihe River Basin of arid northwestern China is discussed in the paper. Under conditions of arid climate and unreasonable human activities, the processes of overuse of water resources by people in the basin have significant effects on the different kinds of environmental degradation, which include water environmental changes, land desertification and salinization, and vegetation degeneration. Furthermore, some proposals for environmental degradation research and control in the study site are put forward.
Perspectives of different parties involved in the selection of capital equipment for sewerage and wastewater‐associated infrastructure investment projects
Authors: Faram M. G. , Saker J. M. , Story V. M.
Published: December 2006

This paper presents a study of UK water industry capital equipment selection, focusing in particular on behaviour in relation to sewerage/combined sewer overflow (CSO) and wastewater‐associated investment activities. Valuable new insights into the perspectives of different parties, including water service providers (WSPs), design consultants and construction companies, are provided. Through analysis of over 200 questionnaires, equipment selection is confirmed to be multi‐organisational, with different parties dominating different roles. Among the selection criteria, service and quality are found to be valued as parameters worth paying for. Relationships and sustainability considerations are also found to be important, although views on whether sustainability benefits will be paid for are divided. While interparty alignment of views is generally good, there are occasional discrepancies at the individual water region level. It is suggested that optimisation might derive from recognising the significance of intangible components of the supplier offering, and managing relationships to allow these to be capitalised upon.
Canine keratoconjunctivitis sicca: disease trends in a review of 229 cases
Authors: Sanchez R. F , Innocent G , Mould J , Billson F. M
Published: December 2006

Objectives:To investigate the disease patterns in dogs with keratoconjunctivitis sicca referred to the University of Glasgow Small Animal Hospital.

Methods:A retrospective study of 229 cases was carried out.

Results:There were 44 breeds in the study, with four breeds, English cocker spaniels, cavalier King Charles spaniels, West Highland white terriers and shih‐tzus, making up 58 per cent of the cases. Among these four breeds, two breed‐dependent disease patterns, one chronic and one acute, were identified. English cocker spaniels and West Highland white terriers had a mean age at onset of clinical signs of five years and one month and five years and six months, respectively, with more females affected than males. Clinical signs consisted predominantly of conjunctival hyperaemia and mucopurulent discharge, with a relatively low incidence of ulcerative keratitis. In contrast, cavalier King Charles spaniels and shih‐tzus showed a more acute disease pattern with a biphasic age distribution at 0 to less than two years of age, and four to less than six and six to less than eight years of age, respectively, with more males affected than females and a significantly higher incidence of ulcerative keratitis in some cases resulting in corneal perforation.

ClinicalSignificance:The study reveals interbreed differences with respect to sex, age and risk of ulcerative keratitis which have not been detailed previously in a referral population.
Improved survival in a retrospective cohort of 28 dogs with insulinoma
Authors: Polton G. A. , White R. N. , Brearley M. J. , Eastwood J. M.
Published: December 2006

Objectives:To determine the survival times for a cohort of dogs with insulinoma and to describe the impact of medical therapy both in non‐surgical cases and in surgical cases following relapse.

Methods:A retrospective study of dogs with insulinoma is presented. The Kaplan‐Meier method is used to evaluate the survival characteristics of this population.

Results:Twenty‐eight dogs were included in the study. The median survival time for all dogs was 547 days. Nineteen patients underwent partial pancreatectomy. The median survival time for this group was 785 days and for those subsequently receiving prednisolone therapy on relapse it was 1316 days. Perioperative complications are discussed.

ClinicalSignificance:Survival times in this study exceed those in other studies published previously. In part, this is explained by an improved remission duration following surgery compared with previous reports. More striking though is the longevity of patients following institution of medical therapy. These data give strong objective support to the role of medical therapy in the management of canine insulinoma, including following relapse after surgically induced remission.
Multiple metastases of thyroid cancer in the cranium and pituitary gland in two dogs
Authors: Tamura S. , Tamura Y. , Suzuoka N. , Ohoka A. , Hasegawa T. , Uchida K.
Published: December 2006

Two dogs, a 14‐year‐old, female American Eskimo dog and a 14‐year‐old, male Maltese dog, were presented with thalamic syndromes, including lowered levels of consciousness, poor postural responses and presence of masses in the neck region. In both dogs, magnetic resonance imaging revealed multiple masses inside the cranium, including the pituitary gland. One dog died from status epilepticus two days after magnetic resonance imaging and the other died two months after magnetic resonance imaging from respiratory failure. These dogs were histopathologically diagnosed with multiple metastases of thyroid cancer occurring inside the cranium, including the pituitary gland. To the authors’ knowledge, this is the first time this tumour pattern has been reported in dogs, but it is possible that it is not uncommon.
Successful closure of feline axillary wounds by reconstruction of the elbow skin fold
Authors: Brinkley C. H.
Published: December 2006

This report describes the successful closure of five chronic feline axillary wounds. The aetiology was known to be forelimb entrapment in a neck collar in three cases and was suspected in the others. Each cat underwent a single surgical procedure during which the wound was debrided, the normal structure of the elbow skin fold was restored and the remaining skin defect was closed primarily. None of the cats had undergone any previous reconstruction attempts. No postoperative complications were observed and the wounds healed uneventfully. Cats have well‐developed elbow skin folds, allowing a wide range of limb motion to occur. Having a forelimb trapped in a neck collar not only creates a wound in the axilla but also disrupts the normal anatomy of the skin fold. This report demonstrates that restoring the elbow skin fold before closing the wound may improve the chances of a successful reconstruction at the first surgical intervention.
Extradural spinal juxtafacet (synovial) cysts in three dogs
Authors: Sale C. S. H. , Smith K. C.
Published: December 2006

Three dogs were presented for investigation of spinal disease and were diagnosed with extradural spinal juxtafacet cysts of synovial origin. Two dogs that were presented with clinical signs consistent with pain in the lumbosacral region associated with bilateral hindlimb paresis were diagnosed using magnetic resonance imaging. Both cysts were solitary and associated with the L6‐7 dorsal articulations; both the dogs had a transitional vertebra in the lumbosacral region. A third dog that was presented with progressive paraparesis localised to T3‐L3 spinal cord segments and compression of the spinal cord at T13‐L1 was diagnosed using myelography. A solitary multiloculated cyst was found at surgery. Decompressive surgery resulted in resolution of the clinical signs in all three dogs. Immunohistological findings indicated that one to two layers of vimentin‐positive cells consistent with synovial origin lined the cysts.
Possible pseudogout in two dogs
Authors: Forsyth S. F. , Thompson K. G. , Donald J. J.
Published: December 2006

Pseudogout, the acute form of calcium pyrophosphate deposition disease, is a common condition in elderly human beings and is characterised by the sudden onset of intense joint pain and synovitis. It is rarely identified in animals but was diagnosed in two dogs that presented with acute lameness and pyrexia. Cytology of the synovial fluid showed a mildly elevated cell count with both non‐degenerate neutrophils and mononuclear cells present. Many of the mononuclear cells and occasional neutrophils contained square or rhomboid‐shaped crystals that were variable in shape and size and weakly birefringent on examination under polarised light. Clinical signs resolved following treatment with prednisolone.
Efficacy and safety of cefovecin (Convenia™) for the treatment of urinary tract infections in dogs
Authors: Passmore C. A. , Sherington J. , Stegemann M. R.
Published: December 2006

Objectives: To determine the efficacy and safety of cefovecin (ConveniaTM; Pfizer Animal Health) in the treatment of urinary tract infections in dogs.

Methods: A multi‐centre, blinded, randomised study was conducted in 129 dogs with urinary tract infections. Cephalexin (RilexineTM) administered twice daily at 15 mg/kg bodyweight orally for 14 days was compared with a single, subcutaneous injection of cefovecin (ConveniaTM) in dogs. The primary efficacy parameter assessed was bacterial elimination of the pretreatment uropathogen.

Results: One hundred and twenty‐nine dogs were included in efficacy assessments. Escherichia coli was eliminated in 90·5 per cent of cefovecin‐treated dogs compared with 52·9 per cent of cephalexin‐treated dogs (P=0·0004). Overall cure rates for dogs with Escherichia coli infections were 79·1 per cent for cefovecin and 36·4 per cent for cephalexin‐treated dogs (P=0·0003). There were no suspected adverse drug reactions attributed to treatment with cefovecin or cephalexin.

Clinical Significance: Cefovecin was shown to be an effective and safe treatment for urinary tract infections.
Male tail streamer length does not predict apparent or genetic reproductive success in North American barn swallowsHirundo rustica erythrogaster
Authors: R. Neuman Colby , J. Safran Rebecca , J. Lovette Irby
Published: December 2006

In the socially monogamous barn swallow, previous studies of individuals in the European subspeciesHirundo r. rusticahave shown that a male's tail streamer length is under strong sexual selection and is positively associated with several measures of reproductive success, including a low probability of being cuckolded by other males. The prominence of these results has led to subsequent experimental and correlational investigations of individuals in the phenotypically divergent subspeciesH. r. erythrogasterin North America, where it has been shown that male tail streamer length is not as strongly associated with reproductive success as in European populations. We examined relationships between male tail streamer length and patterns of: (1) social mate selection and reproductive success, and (2) extra‐pair paternity in 265 progeny of 53 social fathers within a New York barn swallow population. Although tail streamers in this population were sexually dimorphic, male tail streamer length did not predict patterns of mate selection, seasonal reproductive success, or extra‐pair paternity. Moreover, in contrast to the strong positive relationships between paternity and male streamer length in European populations, summarized in this paper, we found no positive relationship between a male's paternity of young in the nest he is attending and his tail streamer length in our study population in New York. Our results further corroborate recent suggestions that the function of sexual signals varies geographically in this species, although we await additional experimental analyses on streamer lengths to understand the maintenance of sexual dimorphism in this trait.
Multiplex Assay for Simultaneous Measurement of Antibodies to Multiple
Plasmodium falciparum
Antigens
Authors: Fouda Genevieve G. , Leke Rose F. G. , Long Carole , Druilhe Pierre , Zhou Ainong , Taylor Diane Wallace , Johnson Armead H.
Published: December 2006

Antibodies to
Plasmodium falciparum
are classically measured using the enzyme-linked immunosorbent assay (ELISA). Although highly sensitive, this technique is labor-intensive when large numbers of samples must be screened against multiple antigens. The suspension array technology (SAT) might be an alterative to ELISA, as it allows measurement of antibodies against multiple antigens simultaneously with a small volume of sample. This study sought to adapt the new SAT multiplex system for measuring antibodies against nine malarial vaccine candidate antigens, including recombinant proteins from two variants of merozoite surface protein 1, two variants of apical merozoite antigen 1, erythrocyte binding antigen 175, merozoite surface protein 3, and peptides from the circumsporozoite protein, ring erythrocyte surface antigen, and liver-stage antigen 1. Various concentrations of the antigens were coupled to microspheres with different spectral addresses, and plasma samples from Cameroonian adults were screened by SAT in mono- and multiplex formats and by ELISA. Optimal amounts of protein required to perform the SAT assay were 10- to 100-fold less than that needed for ELISA. Excellent agreement was found between the single and multiplex formats (
R
≥ 0.96), even when two variants of the same antigen were used. The multiplex assay was rapid, reproducible, required less than 1 μl of plasma, and had a good correlation with ELISA. Thus, SAT provides an important new tool for studying the immune response to malaria rapidly and efficiently in large populations, even when the amount of plasma available is limited, e.g., in studies of neonates or finger-prick blood.
Bovine Immune Response to Shiga-Toxigenic
Escherichia coli
O157:H7
Authors: Hoffman Mark A. , Menge Christian , Casey Thomas A. , Laegreid William , Bosworth Brad T. , Dean-Nystrom Evelyn A.
Published: December 2006

Although cattle develop humoral immune responses to Shiga-toxigenic (Stx
+
)
Escherichia coli
O157:H7, infections often result in long-term shedding of these human pathogenic bacteria. The objective of this study was to compare humoral and cellular immune responses to Stx
+
and Stx
−
E. coli
O157:H7. Three groups of calves were inoculated intrarumenally, twice in a 3-week interval, with different strains of
E. coli
: a Stx2-producing
E. coli
O157:H7 strain (Stx2
+
O157), a Shiga toxin-negative
E. coli
O157:H7 strain (Stx
−
O157), or a nonpathogenic
E. coli
strain (control). Fecal shedding of Stx2
+
O157 was significantly higher than that of Stx
−
O157 or the control. Three weeks after the second inoculation, all calves were challenged with Stx2
+
O157. Following the challenge, levels of fecal shedding of Stx2
+
O157 were similar in all three groups. Both groups inoculated with an O157 strain developed antibodies to O157 LPS. Calves initially inoculated with Stx
−
O157, but not those inoculated with Stx2
+
O157, developed statistically significant lymphoproliferative responses to heat-killed Stx2
+
O157. These results provide evidence that infections with STEC can suppress the development of specific cellular immune responses in cattle, a finding that will need to be addressed in designing vaccines against
E. coli
O157:H7 infections in cattle.
Development of Novel Immunoglobulin G (IgG), IgA, and IgM Enzyme Immunoassays Based on Recombinant Puumala and Dobrava Hantavirus Nucleocapsid Proteins
Authors: Meisel Helga , Wolbert Anne , Razanskiene Ausra , Marg Andreas , Kazaks Andris , Sasnauskas Kestutis , Pauli Georg , Ulrich Rainer , Krüger Detlev H.
Published: December 2006

Human infections with Asian and European hantaviruses can result in hemorrhagic fever with renal syndromes of differing severities characterized by renal dysfunction and sometimes by pulmonary symptoms. For the serological detection of human infections by hantaviruses relevant for Europe, we developed monoclonal antibody capture immunoglobulin G (IgG) and IgA enzyme-linked immunosorbent assays (ELISAs) based on yeast-expressed nucleocapsid proteins of Puumala and Dobrava hantaviruses. Moreover, for diagnosis of acute infections, μ-capture IgM ELISAs were established with nucleocapsid proteins expressed in
Drosophila melanogaster
Schneider S2 cells. The cutoff values of the ELISAs were determined by investigation of up to 500 human anti-hantavirus-negative serum samples. The specificities of the Puumala and Dobrava virus-specific IgM, IgA, and IgG ELISAs were found to be 100%. The sensitivities of these ELISAs were determined to be 100% with panels of characterized anti-Puumala or anti-Dobrava virus-positive human serum samples. In most cases, Puumala and Dobrava virus infections could be differentiated by ELISA reactivity alone, i.e., endpoint titration with homologous and heterologous antigens.
Diagnosis of Paracoccidioidomycosis by Detection of Antigen and Antibody in Bronchoalveolar Lavage Fluids
Authors: Marques-da-Silva Silvia Helena , Colombo Arnaldo Lopes , Blotta Maria Heloisa Souza Lima , Queiroz-Telles Flávio , Balthazar Alípio Barbosa , Lopes José Daniel , de Camargo Zoilo Pires
Published: December 2006

Paracoccidioidomycosis (PCM) is a systemic infection caused by the fungus
Paracoccidioides brasiliensis
and is believed to be the leading cause of fungal pulmonary infection. In this study, we used an inhibition enzyme-linked immunosorbent assay to diagnose pulmonary PCM based on the detection of 43-kDa and 70-kDa molecules in bronchoalveolar lavage fluids. The results were compared with results obtained by classical methods for antibody detection.
Use of the Immunodominant 18-Kilodalton Small Heat Shock Protein as a Serological Marker for Exposure to
Mycobacterium ulcerans
Authors: Diaz Diana , Döbeli Heinz , Yeboah-Manu Dorothy , Mensah-Quainoo Ernestina , Friedlein Arno , Soder Nicole , Rondini Simona , Bodmer Thomas , Pluschke Gerd
Published: December 2006

While it is well established that proximity to wetlands is a risk factor for contracting Buruli ulcer, it is not clear what proportion of a population living in an area where the etiologic agent,
Mycobacterium ulcerans
, is endemic is actually exposed to this disease. Immunological cross-reactivity among mycobacterial species complicates the development of a specific serological test. Among immunodominant proteins recognized by a panel of anti-
M. ulcerans
monoclonal antibodies, the
M. ulcerans
homologue of the
M. leprae
18-kDa small heat shock protein (shsp) was identified. Since this shsp has no homologues in
M. bovis
and
M. tuberculosis
, we evaluated its use as a target antigen for a serological test. Anti-18-kDa shsp antibodies were frequently found in the sera of Buruli ulcer patients and of healthy household contacts but rarely found in controls from regions where the infection is not endemic. The results indicate that only a small proportion of
M. ulcerans
-infected individuals contract the clinical disease.
Clostridium perfringens
Antigens Recognized by Broiler Chickens Immune to Necrotic Enteritis
Authors: Kulkarni R. R. , Parreira V. R. , Sharif S. , Prescott J. F.
Published: December 2006

Little is known about immunity to necrotic enteritis (NE) in chickens. A recent study of broiler chickens showed that protection against NE was associated with infection-immunization with virulent but not with avirulent
Clostridium perfringens
.In the current study, six secreted antigenic proteins unique to virulent
C. perfringens
that reacted to serum antibodies from immune birds were identified by mass spectrophotometry; three of these proteins are part of the VirR-VirS regulon.
Use of Internally Controlled Real-Time Genome Amplification for Detection of Variola Virus and Other Orthopoxviruses Infecting Humans
Authors: Fedele C. G. , Negredo A. , Molero F. , Sánchez-Seco M. P. , Tenorio A.
Published: December 2006

Smallpox, once a devastating disease caused by
Variola virus
, a member of the
Orthopoxvirus
genus, was eradicated in 1980. However, the importance of variola virus infections has been stressed widely in the last few years, particularly following recent social events in the world. Today, variola virus is considered to be one of the most significant agents with potential use as a biological weapon. In this study we developed an internally controlled real-time PCR assay for rapid detection and simultaneous differentiation of variola virus from other orthopoxviruses. The assay is based on TaqMan 3′-minor groove binder (MGB) chemistry and uses generic primers, designed in highly conserved genomic regions of the
crmB
gene, and three TaqMan MGB probes designed to identify orthopoxviruses, variola virus, and an internal control. The results obtained suggest that the assay is rapid, sensitive, specific, and suitable for the generic detection of orthopoxviruses and the identification of variola virus and avoids false-negative results in a single reaction tube.
Neurologic Disease in Captive Lions (Panthera leo) with Low-Titer Lion Lentivirus Infection
Authors: Brennan Greg , Podell Michael D. , Wack Raymund , Kraft Susan , Troyer Jennifer L. , Bielefeldt-Ohmann Helle , VandeWoude Sue
Published: December 2006

Lion lentivirus (LLV; also known as feline immunodeficiency virus of lion,Panthera leo[FIVPle]) is present in free-ranging and captive lion populations at a seroprevalence of up to 100%; however, clinical signs are rarely reported. LLV displays up to 25% interclade sequence diversity, suggesting that it has been in the lion population for some time and may be significantly host adapted. Three captive lions diagnosed with LLV infection displayed lymphocyte subset alterations and progressive behavioral, locomotor, and neuroanatomic abnormalities. No evidence of infection with other potential neuropathogens was found. Antemortem electrodiagnostics and radiologic imaging indicated a diagnosis consistent with lentiviral neuropathy. PCR was used to determine a partial lentiviral genomic sequence and to quantify the proviral burden in eight postmortem tissue specimens. Phylogenetic analysis demonstrated that the virus was consistent with the LLV detected in other captive and free-ranging lions. Despite progressive neurologic signs, the proviral load in tissues, including several regions of the brain, was low; furthermore, gross and histopathologic changes in the brain were minimal. These findings suggest that the symptoms in these animals resulted from nonspecific encephalopathy, similar to human immunodeficiency virus, FIV, and simian immunodeficiency virus (SIV) neuropathies, rather than a direct effect of active viral replication. The association of neuropathy and lymphocyte subset alterations with chronic LLV infection suggests that long-term LLV infection can have detrimental effects for the host, including death. This is similar to reports of aged sootey mangabeys dying from diseases typically associated with end-stage SIV infection and indicates areas for further research of lentiviral infections of seemingly adapted natural hosts, including mechanisms of host control and viral adaptation.
Feasibility of Freeze-Dried Sera for Serological and Molecular Biological Detection of Hepatitis B and C Viruses
Authors: Ohishi Waka , Fujiwara Saeko , Suzuki Gen , Kishi Takeshi , Sora Misae , Matsuura Shinsuke , Hakoda Masayuki , Tatsukawa Yoshimi , Yamada Michiko , Chayama Kazuaki
Published: December 2006

We compared hepatitis B virus (HBV) surface antigen, anti-hepatitis C virus (HCV) antibody, and HCV RNA quantification in frozen and freeze-dried serum samples to assess the usefulness of freeze-dried sera for detection of HBV and HCV. The results indicated that freeze-dried sera as well as frozen sera can be useful for serological and molecular biological analyses of HBV and HCV.
Characterization of aBrucellasp. Strain as a Marine-Mammal Type despite Isolation from a Patient with Spinal Osteomyelitis in New Zealand
Authors: McDonald W. L. , Jamaludin R. , Mackereth G. , Hansen M. , Humphrey S. , Short P. , Taylor T. , Swingler J. , Dawson C. E. , Whatmore A. M. , Stubberfield E. , Perrett L. L. , Simmons G.
Published: December 2006

Naturally acquired infection of humans with a marine mammal-associatedBrucellasp. has only been reported once previously in a study describing infections of two patients from Peru. We report the isolation and characterization of a strain ofBrucellafrom a New Zealand patient that appears most closely related to strains previously identified from marine mammals. The isolate was preliminarily identified asBrucella suisusing conventional bacteriological tests in our laboratory. However, the results profile was not an exact match, and the isolate was forwarded to four international reference laboratories for further identification. The reference laboratories identified the isolate as eitherB. suisorB. melitensisby traditional bacteriological methods in three laboratories and by a molecular test in the fourth laboratory. Molecular characterization by PCR, PCR-restriction fragment length polymorphism, and DNA sequencing of thebp26gene; IS711; theompgenesomp25,omp31,omp2a, andomp2b; IRS-PCR fragments I, III, and IV; and five housekeeping gene fragments was conducted to resolve the discrepant identification of the isolate. The isolate was identified to be closely related to aBrucellasp. originating from a United States bottlenose dolphin (Tursiops truncatus) and common seals (Phoca vitulina).
Specific Primers for Rapid Detection ofMicrosporum audouiniiby PCR in Clinical Samples
Authors: Roque H. D. , Vieira R. , Rato S. , Luz-Martins M.
Published: December 2006

This report describes application of PCR fingerprinting to identify common species of dermatophytes using the microsatellite primers M13, (GACA)4, and (GTG)5. The initial PCR analysis rendered a specific DNA fragment forMicrosporum audouinii, which was cloned and sequenced. Based on the sequencing data of this fragment, forward (MA_1F) and reverse (MA_1R) primers were designed and verified by PCR to establish their reliability in the diagnosis ofM. audouinii. These primers produced a singular PCR band of 431 bp specific only to strains and isolates ofM. audouinii, based on a global test of 182 strains/isolates belonging to 11 species of dermatophytes. These findings indicate these primers are reliable for diagnostic purposes, and we recommend their use in laboratory analysis.
Predominance of Clones Carrying Panton-Valentine Leukocidin Genes among Methicillin-Resistant
Staphylococcus aureus
Strains Isolated in Japanese Hospitals from 1979 to 1985
Authors: Ma Xiao Xue , Ito Teruyo , Chongtrakool Piriyaporn , Hiramatsu Keiichi
Published: December 2006

We examined 97 strains of methicillin-resistant
Staphylococcus aureus
(MRSA) isolated between 1979 and 1985, the period of time when the appearance of MRSA strains increased, and we determined that these strains are distinct from the MRSA clones predominating in today's Japanese hospitals. Type IV staphylococcal cassette chromosome
mec
(SCC
mec
) strains were the most frequent, comprising 53.6% of all strains, followed by type I (22.7%) and type II (21.6%) SCC
mec
strains. Among the type IV SCC
mec
strains, the frequencies of two new subtypes, type IV.3 (IVc) and type IV.4 (IVd), were very high, comprising 38.1 and 10.3% of all strains, respectively. Forty-four of the 97 strains (45.3%) were Panton-Valentine leukocidin (PVL) positive. Among the PVL-positive strains, sequence type 30 (ST30)-SCC
mec
type IV strains producing type 4 coagulase were the most frequent. This is in striking contrast to the MRSA strains isolated in the 1990s, most of which were ST5-SCC
mec
type II strains producing type 2 coagulase and positive for the toxic shock syndrome toxin 1 gene. We also identified a new PVL-carrying prophage lysogenized in a type IV.3 SCC
mec
strain, 81/108. φ108PVL was distinct from the three extant PVL-carrying phages and was presumed to be carried by ST30-type IV.3 SCC
mec
strains isolated in Japan. These results provide genetic bases for the transition of MRSA clones in Japan that is commonly considered the transition from coagulase type 4 MRSA strains to coagulase type 2 MRSA strains. The results also suggested that MRSA strains that predominated between 1979 and 1985 were generated from PVL-positive methicillin-susceptible
S. aureus
strains through the integration of SCC
mec
elements.
Phylogenetic Analysis of “CandidatusMycoplasma turicensis” Isolates from Pet Cats in the United Kingdom, Australia, and South Africa, with Analysis of Risk Factors for Infection
Authors: Willi Barbara , Tasker Séverine , Boretti Felicitas S. , Doherr Marcus G. , Cattori Valentino , Meli Marina L. , Lobetti Remo G. , Malik Richard , Reusch Claudia E. , Lutz Hans , Hofmann-Lehmann Regina
Published: December 2006

Two hemotropic mycoplasmas have been recognized in cats,Mycoplasma haemofelisand “CandidatusMycoplasma haemominutum.” We recently described a third feline hemoplasma species, designated “CandidatusMycoplasma turicensis,” in a Swiss cat with hemolytic anemia. This isolate induced anemia after experimental transmission to two specific-pathogen-free cats and analysis of the 16S rRNA gene revealed its close relationship to rodent hemotropic mycoplasmas. The agent was recently shown to be prevalent in Swiss pet cats. We sought to investigate the presence and clinical importance of “CandidatusMycoplasma turicensis” infection in pet cats outside of Switzerland and to perform the molecular characterization of isolates from different countries. A “CandidatusMycoplasma turicensis”-specific real-time PCR assay was applied to blood samples from 426 United Kingdom (UK), 147 Australian, and 69 South African pet cats. The 16S rRNA genes of isolates from different countries were sequenced and signalment and laboratory data for the cats were evaluated for associations with “CandidatusMycoplasma turicensis” infection. Infections were detected in samples from UK, Australian, and South African pet cats. Infection was associated with the male gender, and “CandidatusMycoplasma haemominutum” andM. haemofeliscoinfection. Coinfected cats exhibited significantly lower packed cell volume (PCV) values than uninfected cats. Phylogenetic analyses revealed that some Australian and South African “CandidatusMycoplasma turicensis” isolates branched away from the remaining isolates. In summary, “CandidatusMycoplasma turicensis” infection in pet cats exists over a wide geographical area and significantly decreased PCV values are observed in cats coinfected with other feline hemoplasmas.
Phaeoacremonium krajdenii
, a Cause of White Grain Eumycetoma
Authors: Hemashettar B. M. , Siddaramappa B. , Munjunathaswamy B. S. , Pangi A. S. , Pattan Jayashree , Andrade A. T. , Padhye A. A. , Mostert Lizel , Summerbell R. C.
Published: December 2006

We describe the first case of white grain pedal eumycetoma caused by
Phaeoacremonium krajdenii
in a 41-year-old man from Goa, India. Based on histological examination of biopsy tissue showing serpentine granules, a culture of the granules yielding phaeoid fungal colonies, and morphological characteristics and sequence comparison of the partial β-tubulin gene with the ex-type isolate of
P. krajdenii
, the causal agent was identified as
P. krajdenii
.
Vulvovaginal-Swab or First-Catch Urine Specimen To Detect
Chlamydia trachomatis
in Women in a Community Setting?
Authors: Skidmore Sue , Horner Paddy , Herring Alan , Sell Joanne , Paul Ian , Thomas Jane , Caul E. Owen , Egger Matthias , McCarthy Anne , Sanford Emma , Salisbury Chris , Macleod John , Sterne Jonathan A. C. , Low Nicola
Published: December 2006

Screening for chlamydia in women is widely recommended. We evaluated the performance of two nucleic acid amplification tests for detecting
Chlamydia trachomatis
in self-collected vulvovaginal-swab and first-catch urine specimens from women in a community setting and a strategy for optimizing the sensitivity of an amplified enzyme immunoassay on vulvovaginal-swab specimens. We tested 2,745 paired vulvovaginal-swab and urine specimens by PCR (Roche Cobas) or strand displacement amplification (SDA; Becton Dickinson). There were 146 women infected with chlamydia. The assays detected 97.3% (95% confidence interval [CI], 93.1 to 99.2%) of infected patients with vulvovaginal-swab specimens and 91.8% (86.1 to 95.7%) with urine specimens. We tested 2,749 vulvovaginal-swab specimens with both a nucleic acid amplification test and a polymer conjugate-enhanced enzyme immunoassay with negative-gray-zone testing. The relative sensitivities obtained after retesting specimens in the negative gray zone were 74.3% (95% CI, 62.8 to 83.8%) with PCR and 58.3% (95% CI, 46.1 to 69.8%) with SDA. In community settings, both vulvovaginal-swab and first-catch urine specimens from women are suitable substrates for nucleic acid amplification tests, but enzyme immunoassays, even after negative-gray-zone testing, should not be used in screening programs.
Comparison of Mannitol Salt Agar and Blood Agar Plates for Identification and Susceptibility Testing of
Staphylococcus aureus
in Specimens from Cystic Fibrosis Patients
Authors: Sharp Susan E. , Searcy Cindy
Published: December 2006

Antimicrobial susceptibilities of
Staphylococcus aureus
strains can be determined accurately by using isolates from mannitol salt agar, and yellow isolates on mannitol salt agar at quantities of >1+ can be reported as
S. aureus
. These methods decrease the time to identification/antimicrobial susceptibility testing of
S. aureus
and decrease costs through eliminating additional testing.
Rapid Simultaneous Detection by Real-Time PCR of Cytomegalovirus UL97 Mutations in Codons 460 and 520 Conferring Ganciclovir Resistance
Authors: Göhring Katharina , Mikeler Elfriede , Jahn Gerhard , Hamprecht Klaus
Published: December 2006

Ganciclovir (GCV) resistance is an emerging problem for transplant recipients. A sensitive and rapid real-time PCR approach for simultaneous and semiquantitative detection of human cytomegalovirus (HCMV) UL97 mutations in codons 460/520 was established by LightCycler and confirmed by restriction fragment length polymorphism and sequencing. Results from HCMV laboratory strains were compared with results from 11 GCV-resistant clinical isolates.
Optimizing Antigen Cocktails for Detection ofMycobacterium bovisin Herds with Different Prevalences of Bovine Tuberculosis: ESAT6-CFP10 Mixture Shows Optimal Sensitivity and Specificity
Authors: Aagaard C. , Govaerts M. , Meikle V. , Vallecillo A. J. , Gutierrez-Pabello J. A. , Suarez-Güemes F. , McNair J. , Cataldi A. , Espitia C. , Andersen P. , Pollock J. M.
Published: December 2006

Bovine tuberculosis is a major problem in many countries; hence, new and better diagnostic tools are urgently needed. In this work, we have tested ESAT6, CFP10, PE13, PE5, MPB70, TB10.4, and TB27.4 for their potentials as diagnostic markers in field animals from Northern Ireland, Mexico, and Argentina, regions with low, medium, and high prevalences of bovine tuberculosis, respectively. At all three sites, ESAT6 and CFP10 were superior diagnostic antigens, while their combination performed even better at the two sites where the combination was tested, providing the best coverage for the detection of diseased populations. The high sensitivity in the skin test reactor groups, combined with the high specificity in the tuberculosis-free groups, indicated that a diagnosis could correctly be made for 85% of the infected animals, based on their responses to these two antigens. Furthermore, TB10.4, PE13, and PE5 have the potential to supplement ESAT6 and CFP10 in a future five-component diagnostic cocktail.