bluuebunny/crossref_metadata_2025 huggingface dataset:
Feeding habits of the exotic black bullheadAmeiurus melas(Rafinesque) in the Iberian Peninsula: first evidence of direct predation on native fish species
Authors: Leunda P. M. , Oscoz J. , Elvira B. , Agorreta A. , Perea S. , Miranda R.
Published: July 2008

The feeding ecology of the exotic invasive black bullheadAmeiurus melaswas conducted in the Iberian Peninsula for the first time. Dietary analysis based on the stomach contents of individuals caught in several Iberian basins was carried out as a first step to evaluate its potential threat for the native Iberian ichthyofauna. Aquatic macroinvertebrates (mainly Chironomidae) dominated the black bullhead’s diet in all size‐classes and sites, irrespective of natural riverine or artificial lentic habitats. Secondary prey items were responsible for the observed between‐sites (microcrustaceans in artificial lentic habitat; oligochaeta and caddisfly larvae in natural riverine habitats) and ontogenetic diet differences (from microcrustaceans to larger prey). These diet variations were also detected in trophic diversity values and feeding strategy plots. Black bullheads consumed plant material, terrestrial prey and co‐occurring fish species (native or exotic) and thus they could be considered as generalist or opportunistic, foraging on the most abundant and available prey. There was no positive relationship between black bullhead size (total length) and fish prey size, probably indicating piscivory on dead or dying vulnerable fishes as well as predation on smaller‐sized active fishes. The results showed that the black bullhead could negatively affect native Iberian ichthyofauna throughout direct predation and competition. Aspects of potential conservation and management implications of fishes resulting from the undesirable presence of the black bullhead in Iberian water bodies are discussed.
Larval development of Gobius xanthocephalus with genetic validation of larval identification
Authors: Monteiro J. , Borges R. , Robalo J. , Almada V. C. , Henriques S. , Gonçalves E. J.
Published: July 2008

Larval development of Gobius xanthocephalus with descriptions of the ontogeny of the main structures, changes in pigmentation patterns and allometric relationships are presented for the first time. No previous descriptions of G. xanthocephalus larvae are available and therefore field collected larvae were compared with larvae obtained under controlled conditions. The DNA of adults, juveniles and larvae was compared in order to validate identifications. The overall developmental sequence obtained agrees with those described for other gobies. Pigmentation patterns were, however, distinct from those of other sympatric Gobiidae allowing a clear identification of G. xanthocephalus larvae. Otolith microstructure analysis revealed a linear age and body length relationship, with an estimated larval growth rate of 0·28 mm day−1.
Myotube production in fast myotomal muscle is switched‐off at shorter body lengths in male than female Atlantic halibutHippoglossus hippoglossus(L.) resulting in a lower final fibre number
Authors: Hagen Ø , Vieira V. L. A , Solberg C , Johnston I. A
Published: July 2008

A sampling method is described to determine accurately the number of fast myotomal muscle fibres (NF) in a large flatfish species, the Atlantic halibutHippoglossus hippoglossus. An unusual feature of the fast myotomal muscle is the presence of internalized strips of slow muscle fibres. In fish of 1·5–3·5 kg (n= 24), the total cross‐sectional area (ATC) of fast muscle was 18% greater in the dorsal than ventral myotomal compartments (P< 0·05), whereas there was no significant difference between left‐ and right‐hand sides of the body. Due the bilateral asymmetry, muscle blocks (5 × 5 × 5 mm) were prepared to systematically sample each myotomal quadrant (dorsal, ventral, left‐ and right‐side) and the diameters of 150 fast fibres measured per block. Smooth non‐parametric probability functions were fitted to a minimum of 800 measurements of fibre diameter per quadrant (n= 5). There were no significant differences in the distribution of muscle fibre diameters between myotomal compartments and thereforeNFcould be estimated from a single quadrant. The number of blocks required to estimateNFwith a repeatability of ±2·5% increased from six at 300 g body mass to 17 at 96·5 kg, caused by variation within and between blocks. Gompertz curves were fitted to measurements of fibre number and fork length (LF). The estimated final fibre number was 8·96 × 105(7·99–9·94 × 105, 95% CI) for males and 1·73 × 106(1·56–1·90 × 106, 95% CI) for female fish. The estimatedLFfor cessation of fibre recruitment in the fast muscle of female fish (1775 mm) was almost twice that in males (810 mm), reflecting their greater ultimate body size.
Juvenile growth and aggression in diploid and triploid Chinook salmon Oncorhynchus tshawytscha (Walbaum)
Authors: Garner S. R. , Madison B. N. , Bernier N. J. , Neff B. D.
Published: July 2008

Multilocus heterozygosity, aggressive and feeding behaviour, plasma cortisol levels and growth rate were evaluated among three groups of juvenile Chinook salmon Oncorhynchus tshawytscha: diploid, triploid and mixed groups of diploid and triploid fish. There was no difference between diploid and triploid fish in measurements of heterozygosity calculated using seven microsatellite loci, and these measurements did not correlate with performance measurements including feeding rate and growth rate. Aggression trials that examined small groups of fish revealed that after 4 days together in tanks, triploid fish were significantly less aggressive during feeding than diploid fish or fish in mixed groups. At the end of the trials, however, plasma cortisol levels did not differ among the three groups. Thirty‐day growth trials in duplicate tanks of 60 fish revealed no difference in growth rate among diploid, triploid and mixed groups, but plasma cortisol levels were significantly lower in triploid fish than in either diploid fish or the mixed fish. Overall, independent of the above differences in aggressive behaviour and cortisol levels, these results suggest similar performance in diploid and triploid Chinook salmon, and thus provide support for the viability of triploid Chinook salmon culture in commercial aquaculture.
Reproductive strategy of leafscale gulper shark Centrophorus squamosus and the Portuguese dogfish Centroscymnus coelolepis on the Portuguese continental slope
Authors: Figueiredo I. , Moura T , Neves A , Gordo L. S
Published: July 2008

Recent data are presented on leafscale gulper shark Centrophorus squamosus and Portuguese dogfish Centroscymnus coelolepis collected during an extended sampling programme carried out at mainland Portuguese landing ports. Although there are some features common to all deep‐water squaliform sharks, the two species have different reproductive strategies: C. squamosus has a lower fecundity and larger total length (LT) at first maturity than C. coelolepis. Despite the scarcity of pregnant C. squamosus in the samples, the LT at birth was estimated at c. 440 mm. For C. coelolepis the results suggest the existence of a non‐annual reproductive cycle with two breeding seasons in each year.
Recovery of Telestes ukliva, an endemic species from the Cetina River, Croatia (Cypriniformes, Cyprinidae)
Authors: Zanella D. , Mrakovčić M. , Mustafić P. , ĆAleta M.
Published: July 2008

Telestes ukliva, endemic to the Cetina River in Croatia and reported as extinct in the 1990s, was rediscovered in 1997. Earlier authors misidentified T. ukliva as the alien Telestes muticellus and erroneously considered T. ukliva to be extinct.
Elevated red myotomal muscle temperatures in the most basal tuna species, Allothunnus fallai
Authors: Sepulveda C. A. , Dickson K. A. , Bernal D. , Graham J. B.
Published: July 2008

The present study tested the hypothesis that Allothunnus fallai can elevate its slow‐oxidative red myotomal muscle (RM) temperature. Measurements on 30 A. fallai (750–850 mm fork length) captured by hook and line off the coast of southern New Zealand revealed that RM temperatures are elevated by mean ±s.d. 8·1 ± 1·3° C (range 6·7–10·0° C) above the mean ±s.d. ambient sea surface temperature 15·3 ± 0·8° C (range 14·3 to 16·4° C). These data provide evidence that the vascular modifications to the central circulation of A. fallai act as a counter‐current heat exchanger and that RM heat conservation is a character state present in all extant tuna species.
Ejaculate expenditure and timing of gamete release in rainbow trout Oncorhynchus mykiss
Authors: Fitzpatrick J. L. , Liley N. R.
Published: July 2008

Using a novel methodology, natural ejaculate volumes of 14 male rainbow trout Oncorhynchus mykiss were compared when males were housed with one female (absence of a rival male) or with another male and one female (rival male present). Contrary to theoretical predictions, male ejaculate expenditure was not influenced by the presence of a rival male. Male gape duration was positively correlated with the volume of sperm ejaculated. Release of sperm by the male always preceded release of eggs by the female. Analysis of the timing and duration of ejaculation suggests that males may rely on the timing and proximity of gamete release to enhance fertilization success. These results are discussed in the context of sperm competition theory.
Characteristics of sperm of polyploid Prussian carp Carassius gibelio
Authors: Flajšhans M. , Rodina M. , Halačka K. , Vetešník L. , Gela D. , Lusková V. , Lusk S.
Published: July 2008

Wild‐captured 16 diploid, five triploid and one tetraploid Prussian carp Carassius gibelio males produced motile haploid, aneuploid (1·5n) and haploid to aneuploid (> 2n) sperm, respectively, in similar concentration and with the lowest percentage of live spermatozoa in sperm for the tetraploid male (mean ±s.d. 83·03 ± 1·76%; P < 0·05) compared to diploid and triploid males (97·37 ± 1·11% and 96·70 ± 1·45%; P > 0·05).
Two types of allograft‐induced cytotoxic macrophage, one against allografts and the other against syngeneic or allogeneic tumor cells
Authors: Yoneda Yukio , Tashiro‐Yamaji Junko , Kubota Takahiro , Yoshida Ryotaro
Published: July 2008

In the 1990s, based on the results of studies using β2M, CD4 or CD8 knockout mice, several groups reported that the main effector cells responsible for skin or organ allograft rejection were non‐T, non‐NK cells. Similarly, we demonstrated that in an animal model of transplantation of BALB/c (H‐2d) skin onto or Meth A (H‐2d) tumor cells into C57BL/6 (H‐2b) mice, AIM, which expressed iNOS, IL‐12, and IL‐18, were the main effector cells and also that they were cytotoxic against syngeneic tumor cells. Here, we examined whether the same population of macrophages could react with two distinct types of target cell. When BALB/c skin or Meth A tumor cells were transplanted into C57BL/6 mice, cytotoxic activity against the allograft was induced in the transplantation site on days 5–14 and was recovered in non‐adherent cells after a 20‐min incubation in a serum‐coated dish, suggesting the induction of a type of AIM (AIM‐1) in the transplantation site. The AIM‐1‐expressing receptors for H‐2DdKd antigens had no cytotoxic activity against syngeneic tumor cells. In contrast, AIM‐2, which were recovered in the fraction adherent to the serum‐coated dish, exhibited cytotoxic activities against various types of tumor cells, whereas they were inactive toward BALB/c skin. AIM expressed iNOS (AIM‐1 < AIM‐2), IL‐12 (AIM‐1 > AIM‐2), and IL‐18 (AIM‐2 alone) mRNAs. These results indicate that after allografting, two distinct types of cytotoxic AIM were induced in the transplantation site, one against the allografted skin or tumor (AIM‐1) and the other against allogeneic or syngeneic tumor cells (AIM‐2).
Protective effects of farnesol against oral candidiasis in mice
Authors: Hisajima Tatsuya , Maruyama Naho , Tanabe Yuko , Ishibashi Hiroko , Yamada Tsuyoshi , Makimura Koichi , Nishiyama Yayoi , Funakoshi Kengo , Oshima Haruyuki , Abe Shigeru
Published: July 2008

Farnesol is known as a quorum‐sensing molecule for Candida albicans and is recognized to play pathogenic roles in Candida infection. To assess the possible role of farnesol in mucosal C. albicans infection, the effects of farnesol treatment against experimental oral candidiasis in mice were examined. Prednisolone‐pretreated ICR mice were orally infected with C. albicans and 3, 24 and 30 hr later the animals were orally given farnesol. Forty‐eight hr later they were killed for observation. Farnesol treatment in a dose ranging between 1.125 and 9 μmol/mouse showed a protective effect against oral candidiasis in a dose‐dependent manner, at least as estimated by symptom scores of tongues. At 9 μmol/mouse it decreased bodyweight loss. Histological studies of 2.25 μmol/mouse farnesol‐treated animals indicated that farnesol suppressed mycelial growth of C. albicans on the surface of tongues, but microbiological study did not prevent the change of CFU of C. albicans cells not only on tongues but also in feces, kidneys and livers. These results suggest that farnesol has very characteristic roles in protection against mucosal candidiasis.
Synthetic fibril peptide promotes clearance of scrapie prion protein by lysosomal degradation
Authors: Okemoto‐Nakamura Yuko , Yamakawa Yoshio , Hanada Kentaro , Tanaka Keiko , Miura Masami , Tanida Isei , Nishijima Masahiro , Hagiwara Ken'ichi
Published: July 2008

Transmissible spongiform encephalopathies are infectious and neurodegenerative disorders that cause neural deposition of aggregates of the disease‐associated form of PrPSc. PrPSc reproduces by recruiting and converting the cellular PrPC, and ScN2a cells support PrPSc propagation. We found that incubation of ScN2a cells with a fibril peptide named P9, which comprises an intrinsic sequence of residues 167–184 of mouse PrPC, significantly reduced the amount of PrPSc in 24 hr. P9 did not affect the rates of synthesis and degradation of PrPC. Interestingly, immunofluorescence analysis showed that the incubation of ScN2a cells with P9 induced colocalization of the accumulation of PrP with cathepsin D‐positive compartments, whereas the accumulation of PrP in the cells without P9 colocalized mainly with lysosomal associated membrane proteins (LAMP)‐1‐positive compartments but rarely with cathepsin D‐positive compartments in perinuclear regions. Lysosomal enzyme inhibitors attenuated the anti‐PrPSc activity; however, a proteasome inhibitor did not impair P9 activity. In addition, P9 neither promoted the ubiquitination of cellular proteins nor caused the accumulation of LC3‐II, a biochemical marker of autophagy. These results indicate that P9 promotes PrPSc redistribution from late endosomes to lysosomes, thereby attaining PrPSc degradation.
Toxoplasma gondii‐derived heat shock protein 70 induces lethal anaphylactic reaction through activation of cytosolic phospholipase A2 and platelet‐activating factor via Toll‐like receptor 4/myeloid differentiation factor 88
Authors: Fang Hao , Mun Hye‐Seong , Kikumura Akitoshi , Sayama Yusuke , Norose Kazumi , Yano Akihiko , Aosai Fumie
Published: July 2008

Toxoplasma gondii‐derived heat shock protein 70 (T.g.HSP70) was proven to induce IFN‐γ‐dependent lethal anaphylactic reaction in T. gondii‐infected mice through an alternative PAF‐mediated pathway, but not the classical immunoglobulin (Ig)E‐dependent pathway. Although marked IFN‐γ production was observed by CD11b+, CD11c+, CD4+ and CD8+ splenocytes, CD11b+ and CD11c+ cells were shown to be the key effecter cells which generated pro‐inflammatory lipid such as PAF and caused T.g.HSP70‐induced anaphylactic reaction. In the present study, we found that the T.g.HSP70‐induced anaphylactic reaction was not observed in TLR 4‐deficient (−/−) mice, whereas it was observed in WT and TLR2−/− mice. The mRNA expression of PAF‐AH, the main enzyme for PAF degradation, increased in T. gondii‐infected WT and TLR2−/− but not in TLR4−/− mice after T.g.HSP70 injection. Furthermore, phosphorylation of cPLA2, which is the key enzyme for pro‐inflammatory lipid generation, was detected in CD11b+ splenocytes of WT and TLR2−/− mice but not in TLR4−/− mice. Subsequently, cPLA2 activation was suppressed by inhibiting the TLR4‐directed p38 and p44/42 MAPK pathways. However, T.g.HSP70‐induced anaphylactic reaction was observed in TRIF−/− mice, but not in MyD88−/− mice. These findings indicate the cPLA2 activated‐PAF production via TLR4/MyD88‐dependent, but not TRIF‐dependent, signaling pathway in T.g.HSP70‐induced anaphylactic reaction in T. gondii‐infected mice.
Polymerase chain reaction detection of bacterial 16S rRNA gene in human blood
Authors: Moriyama Kosei , Ando Chie , Tashiro Kosuke , Kuhara Satoru , Okamura Seiichi , Nakano Shuji , Takagi Yasumitsu , Miki Takeyoshi , Nakashima Yoshiyuki , Hirakawa Hideki
Published: July 2008

Bacterial 16S ribosomal RNA genes (rDNA) were detected in blood samples from two healthy individuals by PCR under conditions involving 30 cycles that did not produce any visible products from negative control saline. Even from control samples, PCR involving 35–40 cycles yielded visible bands. Major clones detected in the blood samples, but not in control, were the Aquabacterium subgroup, Stenotrophomonas subgroup, Budvicia subgroup, Serratia subgroup, Bacillus subgroup and Flavobacteria subgroup. No clone was located within the bacteroides‐clostridium‐lactobacillus cluster, which is indigenous to gastrointestinal flora.
Effect of potato suberin on Streptomyces scabies proteome
Authors: LAUZIER ANNIE , SIMAO‐BEAUNOIR ANNE‐MARIE , BOURASSA SYLVIE , POIRIER GUY G. , TALBOT BRIAN , BEAULIEU CAROLE
Published: July 2008

Two‐dimensional (2D) PAGE was used to detect proteins induced in Streptomyces scabies by potato suberin, a lipidic plant polymer. Nineteen up‐regulated proteins were excised from 2D gels and analysed by N‐terminal sequencing or tandem mass spectrometry (MS/MS). Four of the up‐regulated proteins could be linked to the bacterial response to stress (AldH, GroES, TerD and LexA). Specific metabolic pathways seemed to be activated in the presence of suberin, as shown by the increased expression of specific transporters and of enzymes related not only to glycolysis, but also to nucleotide and amino acid metabolism. Suberin also appeared to influence secondary metabolism as it also caused the overproduction of the BldK proteins that are known to be involved in differentiation and secondary metabolism.
A requirement for calcium in the caspase‐independent killing of Burkitt lymphoma cell lines by Rituximab
Authors: Daniels Ian , Turzanski Julie , Haynes Andrew P.
Published: July 2008

The therapeutic monoclonal antibody rituximab has previously been shown to kill B cells in a caspase‐independent manner. The signalling pathways underpinning this novel death pathway are unknown. The present study showed that rituximab treatment of Burkitt lymphoma cell lines induced a slow rise in intracellular calcium ([Ca2+]i). This rise was only witnessed in cell lines that were killed by antibody, suggesting a critical role for Ca2+in mediating rituximab‐driven caspase‐independent cell death. Inhibition of the two main intracellular store‐located Ca2+channels, i.e. the ryanodine and inositol‐1,4,5‐triphosphate receptor channels by dantrolene and xestospongen‐c respectively did not prevent the rise in Ca2+seen with rituximab or protect cells from subsequent death. In sharp contrast, inhibition of Ca2+entry via plasma membrane channels with (2‐aminoethoxy) diphenylborate or SKF‐96365 or complete chelation of extracellular Ca2+with ethyleneglycol bis (aminoethylether) tetra‐acetate inhibited the rise in [Ca2+]iand increased cell viability. Together, these data suggest that ligation of the CD20 receptor with rituximab allows a slow sustained influx of Ca2+from the external environment that under certain conditions can lead to cell death.
Reduced level of the spindle checkpoint protein BUB1B is associated with aneuploidy in colorectal cancers
Authors: Burum‐Auensen E. , DeAngelis P. M. , Schjølberg A. R. , Røislien J. , Mjåland O. , Clausen O. P. F.
Published: July 2008

Abstract.  Objectives: The majority of solid human malignancies demonstrate DNA aneuploidy as a consequence of chromosomal instability. We wanted to investigate whether Aurora A, Aurora B, BUB1B and Mad2 were associated with the development of aneuploidy in colorectal adenocarcinomas as suggested by several in vitro studies, and if their protein levels were related to alterations at the corresponding chromosomal loci. Materials and methods: Expression levels of these spindle proteins were investigated by immunohistochemistry using tissue micro‐arrays in a series of DNA aneuploid and diploid colorectal adenocarcinomas previously examined for genomic aberrations by comparative genomic hybridization. Results: All proteins were overexpressed in malignant tissues compared to controls (P < 0.001 for all). BUB1B level was significantly reduced in aneuploid compared to diploid cancers (P = 0.001), whereas expression of the other proteins was not associated with DNA ploidy status. High levels of Aurora A (P = 0.049) and low levels of Aurora B (P = 0.031) were associated with poor prognosis, but no associations were revealed between protein expression and genomic aberration. Conclusions: A significant reduction of BUB1B level was detected in aneuploid compared to diploid colorectal cancers, consistent with earlier studies showing that loss of spindle checkpoint function may be involved in development of DNA aneuploidy. Our data also show that spindle proteins are overexpressed in colorectal cancers, and that expression of the Aurora kinases is associated with prognosis in colorectal cancer.
Exogenous bone marrow cells do not rescue non‐irradiated mice from acute renal tubular damage caused by HgCl2, despite establishment of chimaerism and cell proliferation in bone marrow and spleen
Authors: Fang T.‐C. , Otto W. R. , Jeffery R. , Hunt T. , Alison M. R. , Cook H. T. , Wright N. A. , Poulsom R.
Published: July 2008

Abstract.  Objective: Various studies have shown that bone marrow stem cells can rescue mice from acute renal tubular damage under a conditioning advantage (irradiation or cisplatin treatment) favouring donor cell engraftment and regeneration; however, it is not known whether bone marrow cells (BMCs) can contribute to repair of acute tubular damage in the absence of a selection pressure for the donor cells. The aim of this study was to examine this possibility. Materials and methods: Ten‐week‐old female mice were assigned into control non‐irradiated animals having only vehicle treatment, HgCl2‐treated non‐irradiated mice, HgCl2‐treated non‐irradiated mice infused with male BMCs 1 day after HgCl2, and vehicle‐treated mice with male BMCs. Tritiated thymidine was given 1 h before animal killing. Results: Donor BMCs could not alleviate non‐irradiated mice from acute tubular damage caused by HgCl2, deduced by no reduction in serum urea nitrogen combined with negligible cell engraftment. However, donor BMCs could home to the bone marrow and spleen and display proliferative activity. This is the first report to show that despite no preparative myeloablation of recipients, engrafted donor BMCs can synthesize DNA in the bone marrow and spleen. Conclusions: Exogenous BMCs do not rescue non‐irradiated mice from acute renal tubular damage caused by HgCl2, despite establishment of chimerism and cell proliferation in bone marrow and spleen.
Living on an island: consequences of childhood abuse, attachment disruption and adversity in later life
Authors: Frederick John , Goddard Chris
Published: July 2008

This paper reports on a study which sought to obtain greater knowledge and understanding of the life circumstances of a group of people who had experienced adversity and abuse in childhood. A qualitative research strategy involving in‐depth interviews with individuals was used to help achieve the study's aim. The study found that interviewees detailed multiple risks to their development during childhood and adolescence, including experiences of child abuse, being placed in care and major grief and loss experiences. Most had left home in their mid‐teenage years and had little or no contact with their families of origin. Interviewees had very limited social networks and described difficulties in making and keeping friends. Many reported unstable and often violent personal relationships. As suggested by attachment theory, this study supports the notion of developmental continuities from adverse childhood experiences into later life, particularly in relation to personal and social relationships. Recommendations for a number of suitable methods of intervention and support are outlined.
Social Work Practices: silences and elisions in the plan to ‘transform’ the lives of children ‘looked after’ in England
Authors: Garrett Paul Michael
Published: July 2008

This paper critically examines the government's proposals for the reform of services for children in public care, or ‘looked after’, in England and, more specifically, the proposal to set up Social Work Practices (SWPs, see Note 1). These are envisaged as groups of social workers, commissioned by but independent of local authorities. The changes have been mapped out by a Green Paper Care Matters: Transforming the Lives of Children and Young People in Care published in October 2006, a subsequent working group report, and Care Matters: Time for Change, a White Paper published in June 2007. This was followed, in November, by the publication of the Children and Young Persons’ Bill. It is argued that there are, at least, two issues which could be focused on by those seeking to counter moves to introduce SWPs. First, the enhanced role being given to private sector providers because, it is maintained, public services provided by local authorities are unambiguously ‘failing’ children. Second, the fact that the views of parents and ‘looked after’ children are mostly excluded from the dominant and ‘official’ discourse on SWPs.
Dyadic Developmental Psychotherapy: an evidence‐based treatment for children with complex trauma and disorders of attachment
Authors: Becker‐Weidman Arthur , Hughes Daniel
Published: July 2008

Dyadic Developmental Psychotherapy (DDP) is a family therapy treatment based on Attachment Theory. It is an integration of several approaches, methods and techniques that have a strong evidence base. The approach was originally developed to treat children with disorders of attachment and has been shown to be effective. It has since been more broadly used to help families with a variety of difficulties, including complex trauma. This paper outlines the primary principles and components of DDP and the evidence to support the effectiveness of the components and, therefore, of DDP.
Immunoglobulin E recognition patterns to purified Kiwifruit (Actinidinia deliciosa) allergens in patients sensitized to Kiwi with different clinical symptoms
Authors: Palacin A. , Rodriguez J. , Blanco C. , Lopez‐Torrejon G. , Sánchez‐Monge R. , Varela J. , Jiménez M. A. , Cumplido J. , Carrillo T. , Crespo J. F. , Salcedo G.
Published: July 2008

Background
Green kiwifruit allergy is on the rise. However, no surveys testing purified major kiwi allergens have been carried out in a large population, including both kiwi‐sensitized [skin prick test (SPT)‐positive] and truly kiwi‐allergic patients.

Objective
To isolate major kiwifruit allergens, and to explore their relevance by in vitro and in vivo methods in a large kiwi‐sensitized and ‐allergic population.

Methods
A large group (n=92) of kiwi‐sensitized patients with different clinical symptoms were selected, and double‐blind, placebo‐controlled, food challenges to kiwi were performed in 52 of them. The three major IgE‐binding proteins from kiwifruit extracts were isolated and characterized by N‐terminal amino acid sequencing and molecular size and glycosylation analysis. The allergenic potency of the three kiwi allergens, and of avocado Pers a 1 as a model allergen associated with the latex‐fruit syndrome, was tested by specific IgE quantitation, immunodetection assays and SPTs.

Results
The isolated kiwifruit allergens were identified as actinidin Act d 1, glycosylated thaumatin‐like Act d 2 and a novel 40 kDa glycoprotein designated as Act d 3.02. Specific IgE to each of the three allergens was found in over 60% of sera from kiwi‐sensitized patients, and Act d 1 and Act d 2 induced positive SPT responses in over 50% of the tested patients. A significant link between IgE levels to Act d 1 and Act d 3 and anaphylaxis was uncovered. Avocado Pers a 1 showed an in vitro sensitization prevalence of around 45%, but a low in vivo reactivity.

Conclusion
Act d 1, Act d 2 and Act d 3 are major allergens in the population studied. Severe symptoms after kiwi ingestion are associated with high IgE levels to Act d 1 and Act d 3.
Induction of T‐helper type 2 cytokine release and up‐regulated expression of protease‐activated receptors on mast cells by recombinant American cockroach allergen Per a 7
Authors: Zhang Z. , Zhang H. , Yang H. , Zhang L. , Chen X. , Zheng X. , He S.
Published: July 2008

Background
The cockroach is a major source of indoor allergens, which cause perennial rhinitis and asthma. Per a 7 is one of the major allergens from the American cockroach, eliciting IgE reaction in over 50% of sera from allergic individuals. However, the actions of Per a 7 in the pathogenesis of allergic diseases remain poorly understood.

Objective
The aim of the study was to investigate the effects of Per a 7 on the expression of protease‐activated receptors (PARs) on murine mast cells and the release of T‐helper type 2 (Th2) cytokines from these cells.

Methods
Per a 7 gene was cloned and expressed in Escherichia coli. The purified recombinant Per a 7 (rPer a 7) was used to challenge murine mast cells P815 and the expression of PARs was determined by using real‐time quantitative RT‐PCR and flow cytometry. The release of IL‐4 and IL‐13 was detected with ELISA.

Results
rPer a 7 was solubly expressed in E. coli and the purified rPer a 7 reacted with 75% (six out of eight) of the sera from cockroach‐allergic patients. Following stimulation of rPer a 7, the expression of PAR‐1, PAR‐2, PAR‐3 and PAR‐4 mRNAs on P815 cells increased by 8.7‐, 6.8‐, 14.4‐ and 8.8‐fold, respectively, following 2‐ and 6‐h incubation periods, and the expression of PAR‐1, PAR‐2 and PAR‐4 proteins was enhanced following 16‐h incubation. rPer a 7 provoked approximately up to a 4.8‐ and 2.3‐fold increase in IL‐4 and IL‐13 release from P815 cells following 6‐ and 16‐h incubation periods.

Conclusion
rPer a 7 induced up‐regulation of PAR expression on P815 cells and provoked Th2 cytokines IL‐4 and IL‐13 secretion from these murine mast cells, which implicated that this major cockroach allergen is likely to contribute to the development of cockroach‐related allergic disease.
Toll‐like receptor 4 agonists adsorbed to aluminium hydroxide adjuvant attenuate ovalbumin‐specific allergic airway disease: role of MyD88 adaptor molecule and interleukin‐12/interferon‐γ axis
Authors: Bortolatto J. , Borducchi É. , Rodriguez D. , Keller A. C. , Faquim‐Mauro E. , Bortoluci K. R. , Mucida D. , Gomes E. , Christ A. , Schnyder‐Candrian S. , Schnyder B. , Ryffel B. , Russo M.
Published: July 2008

Background
Epidemiological and experimental data suggest that bacterial lipopolysaccharides (LPS) can either protect from or exacerbate allergic asthma. Lipopolysaccharides trigger immune responses through toll‐like receptor 4 (TLR4) that in turn activates two major signalling pathways via either MyD88 or TRIF adaptor proteins. The LPS is a pro‐Type 1 T helper cells (Th1) adjuvant while aluminium hydroxide (alum) is a strong Type 2 T helper cells (Th2) adjuvant, but the effect of the mixing of both adjuvants on the development of lung allergy has not been investigated.

Objective
We determined whether natural (LPS) or synthetic (ER‐803022) TLR4 agonists adsorbed onto alum adjuvant affect allergen sensitization and development of airway allergic disease. To dissect LPS‐induced molecular pathways, we used TLR4‐, MyD88‐, TRIF‐, or IL‐12/IFN‐γ‐deficient mice.

Methods
Mice were sensitized with subcutaneous injections of ovalbumin (OVA) with or without TLR4 agonists co‐adsorbed onto alum and challenged with intranasally with OVA. The development of allergic lung disease was evaluated 24 h after last OVA challenge.

Results
Sensitization with OVA plus LPS co‐adsorbed onto alum impaired in dose‐dependent manner OVA‐induced Th2‐mediated allergic responses such as airway eosinophilia, type‐2 cytokines secretion, airway hyper‐reactivity, mucus hyper production and serum levels of IgE or IgG1 anaphylactic antibodies. Although the levels of IgG2a, Th1‐affiliated isotype increased, investigation into the lung‐specific effects revealed that LPS did not induce a Th1 pattern of inflammation. Lipopolysaccharides impaired the development of Th2 immunity, signaling via TLR4 and MyD88 molecules and via the IL‐12/IFN‐γ axis, but not through TRIF pathway. Moreover, the synthetic TLR4 agonists that proved to have a less systemic inflammatory response than LPS also protected against allergic asthma development.

Conclusion
Toll‐like receptor 4 agonists co‐adsorbed with allergen onto alum down‐modulate allergic lung disease and prevent the development of polarized T cell‐mediated airway inflammation.
Prior SO2 exposure promotes airway inflammation and subepithelial fibrosis following repeated ovalbumin challenge
Authors: Cai C. , Xu J. , Zhang M. , Chen X‐D. , Li L. , Wu J. , Lai H‐W. , Zhong N‐S.
Published: July 2008

Background
Exposure to allergens or air pollutants often leads to asthma exacerbations associated with aggravation of airway inflammation. Although, repeated allergen challenge often induces chronic allergic airway inflammation (CAAI) and airway remodelling, yet, the effects of brief exposure to air pollutants such as SO2 on development of CAAI and airway remodelling remain to be clarified.

Objective
The aim of the experiment was to investigate the effects of acute neutrophilic airway inflammation induced by brief exposure to SO2 on development of CAAI and subepithelial fibrosis (SEF) in a murine model of asthma.

Methods
Acute airway inflammation was induced by brief exposure to 50 p.p.m. SO2 (1 h/d, 3 days). CAAI and SEF in BALB/c mice were induced by repeated challenge with ovalbumin (OVA) for 5 or 9 weeks with or without prior exposure to SO2. Bronchoalveolar lavage fluid (BALF) eosinophilia as index of CAAI, BALF endothelin‐1 (ET‐1) and TGF‐β1 levels, morphometric evaluation of fibrotic area beneath subbasement membrane and lung hydroxyproline content (Hyp) as indexes of SEF were monitored.

Results
Exposure to SO2 led to acute neutrophilic inflammation and epithelial sloughing with profound elevation of BALF ET‐1. Repeated OVA challenge resulted in CAAI and SEF along with elevation of Hyp, increase of fibrotic area beneath subbasement membrane and elevation of BALF TGF‐β1. Preceding SO2 exposure exaggerated BALF eosinophilia, facilitated and enhanced SEF with more significant elevation of BALF ET‐1 and TGF‐β1 levels compared with OVA‐challenged mice without prior exposure to SO2. The increase of Hyp was positively correlated with elevation of BALF TGF‐β1 during CAAI (r=0.842, P<0.01).

Conclusion
This data demonstrated that SEF developed in parallel with severity and time course of CAAI following repeated OVA challenge. SO2‐induced acute epithelial injury and neutrophilic inflammation could enhance CAAI and promote SEF, probably through overexpression of ET‐1 and TGF‐β1.
Exposure to environmental bacteria may have differing effects on tumour necrosis factor‐α and interleukin‐6‐producing capacity in infancy
Authors: Lappalainen M. H. J. , Roponen M. , Hyvärinen A. , Nevalainen A. , Laine O. , Pekkanen J. , Hirvonen M.‐R.
Published: July 2008

Background
Our previous study showed an association between increased concentration of endotoxin in house dust and elevated IFN‐γ responses in neonates. The impact of other microbial agents on immune responses in infancy is poorly known.

Objective
To examine whether stimulated cytokine responses of mothers and their children are associated with concentrations of other microbial markers in addition to endotoxin in house dust samples.

Methods
Mitogen‐stimulated production of IFN‐γ, IL‐4, IL‐6 and TNF‐α was measured in cord blood and in peripheral blood of mothers (n=29) and their children (n=29) 3 months after birth. Gas chromatography mass spectrometric analysis was applied to measure the concentrations of ergosterol (marker of fungal biomass), muramic acid (indicating the presence of Gram‐positive bacteria) and 3‐hydroxy fatty acids (C10:0–C14:0, indicating the presence of Gram‐negative bacteria) in house dust. Endotoxin was determined with Limulus assay.

Results
Significant mother‐to‐child correlations were observed in stimulated production of TNF‐α and IL‐6 3 months after birth. 3‐hydroxy fatty acid (C10:0–C14:0) levels in bed dust were inversely associated with the production of TNF‐α and IL‐6 in blood samples of mothers and their 3‐month‐old children. High concentrations of muramic acid in floor dust were related to increased production of TNF‐α and IL‐6 at the age of 3 months. In contrast to endotoxin, none of the other microbial markers were significantly associated with enhanced IFN‐γ‐producing capacity from birth to 3 months.

Conclusions
Exposure to Gram‐negative bacteria and their components may be associated with down‐regulated immune responses in early infancy, indicated as an impaired production of pro‐inflammatory cytokines following mitogen stimulation. Gram‐positive bacteria and their constituents seem to have opposite effects. Of the measured markers, exposure to bioactive endotoxin appears to have the strongest impact on T‐helper type 1 responses.
Supplementation with Lactobacillus rhamnosus or Bifidobacterium lactis probiotics in pregnancy increases cord blood interferon‐γ and breast milk transforming growth factor‐β and immunoglobin A detection
Authors: Prescott S. L. , Wickens K. , Westcott L. , Jung W. , Currie H. , Black P. N. , Stanley T. V. , Mitchell E. A. , Fitzharris P. , Siebers R. , Wu L. , Crane J. ,   the Probiotic Study Group
Published: July 2008

Background
This study explored the effects of maternal probiotic supplementation on immune markers in cord blood (CB) and breast milk.

Methods
CB plasma and breast milk samples were collected from a cohort of women who had received daily supplements of either 6 × 109 CFU/day Lactobacillus rhamnosus HN001 (n=34), 9 × 109 CFU/day Bifidobacterium lactis HN019 (n=35) or a placebo (n=36) beginning 2–5 weeks before delivery and continuing for 6 months in lactating women. CB plasma and breast milk (collected at 3–7 days, 3 months and 6 months postpartum) were assayed for cytokines (IL‐13, IFN‐γ, IL‐6, TNF‐α, IL‐10, TGF‐β1) and sCD14. Breast milk samples were also assayed for total IgA.

Results
Neonates of mothers who received a probiotic had higher CB IFN‐γ levels (P=0.026), and a higher proportion had detectable blood IFN‐γ levels, compared with the placebo group (P=0.034), although levels were undetectable in many infants. While this pattern was evident for both probiotics, when examined separately only the L. rhamnosus HN001 group showed statistically significant higher IFN‐γ levels (P=0.030) compared with the placebo group. TGF‐β1 levels were higher in early breast milk (week 1) from the probiotic groups (P=0.028). This was evident for the B. lactis HN019 group (P=0.041) with a parallel trend in the L. rhamnosus HN001 group (P=0.075). Similar patterns were seen for breast milk IgA, which was more readily detected in breast milk from both the B. lactis HN019 (P=0.008) and the L. rhamnosus HN001 group (P=0.011). Neonatal plasma sCD14 levels were lower in the B. lactis HN019 group compared with the placebo group (P=0.041).

Conclusion
The findings suggest that supplementation with probiotics in pregnancy has the potential to influence fetal immune parameters as well as immunomodulatory factors in breast milk.
The effect of lubricant contamination on ThinPrep® (Cytyc) cervical cytology liquid‐based preparations
Authors: Holton T. , Smith D. , Terry M. , Madgwick A. , Levine T.
Published: July 2008

Objectives:  To assess the extent of lubricant use by smear‐takers and the effect of lubricant contamination of ThinPrep® processed cervical cytology samples.

Methods:  All primary care smear‐takers were sent a questionnaire on lubricant type and frequency of use. Fifty cervical cytology samples were then contaminated with incremental amounts of K‐Y® jelly, 50 samples contaminated with incremental amounts of Aquagel® and ten non‐contaminated vials were processed using the ThinPrep® T2000 processor followed by Papanicolaou staining. The morphological appearances of lubricant contamination were described microscopically and formal cell counts performed on all slides.

Results:  Seventy of 94 (74.5%) primary care smear‐takers indicated lubricant use of whom 9/70 (12.8%) used Aquagel® and 61/70 (87.2%) used K‐Y® jelly. K‐Y® jelly appeared as mucoid blue deposits in the slide background whereas Aquagel® appeared as pink stringy background material. Cell counting showed a significant difference between Aquagel® and K‐Y® jelly contaminated slides compared to the original non‐contaminated preparations for all fields and the average fields (P < 0.001) with a significantly higher count for the original non‐contaminated slides than the lubricant contaminated groups.

Conclusion:  Lubricant contamination of ThinPrep® cervical cytology samples may result in reduced cellularity of the subsequent slide. This study provides evidence‐based data to support British Society for Clinical Cytology recommendations for no lubricant use when taking cervical samples.
Immunocytochemical panel for distinguishing carcinoma cells from reactive mesothelial cells in pleural effusions
Authors: Wu G‐P. , Zhang S‐S. , Fang C‐Q. , Liu S‐L. , Wang E‐H.
Published: July 2008

Objective:  The aim of this study was to evaluate the individual and combined diagnostic utility of carcinoembryonic antigen (CEA), cytokeratin 19 fragments (CK19) and HBME‐1 in pleural effusions of patients with lung cancer.

Study design:  CEA, CK19 and HBME‐1 were detected by immunocytochemistry in pleural effusions from patients with lung cancer (86 cases) and without lung cancer (40 cases).

Results:  CEA and CK19 expression were significantly higher in the carcinoma cell group and in three subgrouped as adenocarcinoma (AC), squamous cell carcinoma (SCC) and small cell lung cancer than in the mesothelial cell group, whereas HBME‐1 expression was lower in the former group (P < 0.01). In the subgrouped tumours, CEA expression was higher in AC than in SCC (P < 0.05), whereas HBME‐1 expression was higher in SCC than in AC (P < 0.01). Used alone, CK19 had the highest sensitivity (95.3%) and accuracy (93.7%), whereas CEA had the highest specificity (97.5%). When combinations of antibodies were evaluated together and membrane staining with HBME‐1 taken as a negative outcome, CK19 and HBME‐1 gave a high diagnostic performance: sensitivity of 100.0% and accuracy of 95.2% respectively.

Conclusion:  A panel of CEA, CK19 and HBME‐1 monoclonal antibodies proved to be suitable for distinguishing carcinoma cells from reactive mesothelial cells in pleural effusions.
Paraganglioma‐like medullary thyroid carcinoma: fine needle aspiration cytology features with histological correlation
Authors: Ryska A. , Cap J. , Vaclavikova E. , Dvorakova S. , Bendlova B. , Hovorkova E. , Kohout A.
Published: July 2008

Objectives:  Two cases of an extremely rare paraganglioma‐like variant of medullary thyroid carcinoma (MTC) are reported.

Methods:  The patients were a 65‐year‐old male (case 1) and a 14‐year‐old female (case 2). Unilateral thyroid nodule and homolateral cervical lymphadenopathy was present in case 1; bilateral thyroid nodules were seen in case 2. Fine needle aspiration cytology (FNAC) was performed from thyroid nodules (in both cases) and from a cervical lymph node (in case 1).

Results:  The cytological smears contained predominantly ovoid to spindled epithelial cells arranged in cohesive three‐dimensional clusters with sharp margins; isolated individual cells were seen only rarely. No colloid or other material was present in the background. The tumour cells showed significant nuclear atypia with occasional bizarre and/or binucleated cells. The nuclear chromatin was coarse and granular, sometimes with grooves and intranuclear inclusions. The cytoplasm was inconspicuous. Polygonal or triangular cells, amyloid and azurophillic cytoplasmic granules were absent in both cases. Calcitonin expression was demonstrated in case 2. Histological examination confirmed the paraganglioma variant of MTC in both cases.

Mutation of RET proto‐oncogene in exon 16 (Met918Thr) – germline in case 2 and somatic in case 1 was detected by sequencing of DNA in both cases.

Conclusions:  This is the first description of cytological findings in the paraganglioma‐like variant of MTC. Despite its rarity, it can be reliably diagnosed by FNAC if material for immunocytochemistry is obtained.
Could nuclear matrix protein 22 (NMP22) play a role with urine cytology in screening for bladder cancer? – experience at Kuwait University
Authors: Kapila K. , Kehinde E. O. , Anim J. T. , Mojiminiyi O. A. , Vinsu A. , George S. S. , Al‐Maghrebi M. , Al‐Mulla F.
Published: July 2008

Objectives:  This prospective study was undertaken to evaluate nuclear matrix protein (NMP22) compared to urine cytology in the detection of bladder cancer and also to determine whether indexing suspicious cytology to NMP22 could enhance the clinical utility of cytology.

Methods:  Cytological findings of voided urine collected prior to a cystoscopic biopsy were correlated with urine NMP22 assay in 46 patients attending the urology clinic in Mubarak Al‐Kabeer Hospital. The patients were clinically categorized into newly diagnosed cases of transitional cell carcinoma (TCC), recurrent TCC, TCC in remission and controls.

Results:  Using histological diagnosis as the gold standard the sensitivity and specificity of NMP22 were 78% and 43% respectively and of cases with malignant urine cytology were 30% and 87% respectively. If suspicious and malignant cytology were combined as positive results the sensitivity increased significantly to 87% while the specificity decreased but not significantly to 74%. Suspicious or malignant cytology enhanced by positive NMP22 gave a sensitivity of 70% and specificity of 87% neither of which was significantly different from cytology alone. There were three false positive cases on cytology and 13 false positive cases on NMP22 assay. There were three false negative cytology and five false negative NMP22 cases but only one was false negative for both, resulting in a high sensitivity (96%) but low specificity (30%) if either positive NMP22 or malignant or suspicious cytology was taken as a positive result.

Conclusion:  Combining NMP22 with malignant or suspicious cytological result improved sensitivity for the detection of bladder cancer but with a major decrease in specificity, suggesting a potential role in screening rather than diagnosis.