bluuebunny/crossref_metadata_2025 huggingface dataset:
Atopic dermatitis in young children is associated with impaired interleukin‐10 and interferon‐γ responses to allergens, vaccines and colonizing skin and gut bacteria
Authors: Dunstan J. A. , Hale J. , Breckler L. , Lehmann H. , Weston S. , Richmond P. , Prescott S. L.
Published: October 2005

Background
A significant proportion of children with food allergy and more severe forms of atopic dermatis (AD) go on to develop persistent forms of allergic disease such asthma. Defining immune dysregulation in these children will be of great value in understanding disease pathogenesis.

Objective
In this study we characterized the immune responses of young infants (6–18 months of age) with moderate‐to‐severe AD (a modified SCORAD25) and compared these (n=53) with responses of non‐allergic children with no history of dermatitis or sensitization of the same age (n=20).

Methods
Mononuclear cell cytokine responses to allergens (egg ovalbumin (OVA), β‐lactoglobulin (BLG), house dust mite (HDM)), vaccines (tetanus toxoid (TT), diphtheria toxoid (DT)), intestinal flora (heat‐killed Lactobacillus species (HKLB)), heat‐killed Staphylococcus aureus (HKSA), S. aureus enterotoxin B (SEB) and mitogen (phytohaemaglutinin (PHA)) were compared in children with AD with unaffected children.

Results
Children with AD had significantly lower spontaneous (unstimulated) production of regulatory cytokine IL‐10 (P<0.001), as well as IFN‐γ (P<0.001) and TNF‐α (P<0.001) compared with the unaffected children. After allowing for differences in baseline levels IL‐10 responses to virtually all stimuli (food allergens (P=0.003), vaccines P=0.01, intestinal flora (heat‐killed Lactobacillus species (HKLB), P=0.005) and skin flora (heat‐killed Staphylococcus aureus (HKSA), P=0.003)) were also significantly attenuated in children with AD. The only exception was HDM, to which responses were stronger in children with AD [P=0.05]. Although there were no significant correlations between HDM IgE and HDM cytokine responses at this age, T‐helper type 2 (Th2) IL‐5 (P=0.014) and IL‐13 (P=0.004) responses to HDM were significantly more frequent in the children with AD. However, while children with AD showed significantly attenuated Th1 IFN‐γ responses to food allergens (OVA, P=0.007 and BLG, P<0.001) and vaccines (DT, P=0.008 and TT, P<0.001), these children showed no difference in Th1 IFN‐γ responses to HDM or microbial agents (HKSA and HKLB).

Conclusion
A increase in propensity for Th2 responses to aeroallergens in children with AD is associated with early impaired production of IL‐10 regulatory cytokine to a broad range of environmental stimuli including foods, intestinal flora, S. aureus, and vaccines.
The squamous cell carcinoma antigens as relevant biomarkers of atopic dermatitis
Authors: Mitsuishi K. , Nakamura T. , Sakata Y. , Yuyama N. , Arima K. , Sugita Y. , Suto H. , Izuhara K. , Ogawa H.
Published: October 2005

Background
Although it is thought that both Th1‐ and Th2‐type inflammations are involved in the pathogenesis of atopic dermatitis (AD), it is controversial which immune response is more involved in regulating the clinical severity of AD. We recently found that the squamous cell carcinoma antigens 1 (SCCA1) and SCCA2 are novel biomarkers of bronchial asthma, downstream of IL‐4 and IL‐13.

Objective
We examined whether SCCA1 and SCCA2 could also serve as biomarkers of AD, reflecting its Th2‐type immune responses, and whether the expression level of SCCA was correlated with clinical severity of AD.

Method
We compared the expression of SCCA1 and SCCA2 at the mRNA and protein levels in both involved and uninvolved skin of AD patients and in normal control skin. We next analysed induction of SCCA by IL‐4 or IL‐13 in keratinocytes. Finally, we compared the serum level of SCCA with laboratory parameters reflecting Th2‐type inflammation and clinical severity in AD patients.

Results
SCCA1 and SCCA2 were highly expressed in involved skin of AD patients, compared with their uninvolved skin, at both mRNA and protein levels. SCCA protein was dominantly expressed in suprabasal keratinocytes in the epidermis of AD patients. Either IL‐4 or IL‐13, but not IFN‐γ or TNF, induced production of SCCA in keratinocytes. These result suggest that SCCA is induced in AD skin, probably due to direct actions of IL‐4 and/or IL‐13 on keratinocytes. Serum levels of SCCA were well correlated with eosinophil numbers and serum lactate dehydrogenase levels, and weakly with serum IgE levels, in AD patients. Furthermore, serum levels of SCCA were strongly correlated with clinical severity.

Conclusions
Th2‐type inflammation dominantly regulates the clinical severity of AD, and SCCA is a relevant biomarker of AD, reflecting both Th2‐type inflammation and clinical severity.
Dysfunction of the mitotic:meiotic switch as a potential cause of neoplastic conversion of primordial germ cells
Authors: ADAMAH D. J. B. , GOKHALE P. J. , EASTWOOD D. J. , RAJPERT DE‐MEYTS E. , GOEPEL J. , WALSH J. R. , MOORE H. D. , ANDREWS P. W.
Published: October 2005

Germ cell tumours (GCT) are thought to arise as the result of a defect in early development, probably shortly after arrival of the migrating primordial germ cells (PGC) in the genital ridge when, if in a male genital ridge, the germ cells arrest in mitosis, but in a female genital ridge they enter meiosis. We suggest that dysfunction of the mitotic:meiotic switch, with cells aberrantly co‐expressing functions pertinent to both states, might provide the genetic instability that could initiate tumour development. If this hypothesis is correct, GCT could arise because of disruption in the function of any one of a number of different genes involved in controlling mitosis and meiosis, rather than being dependent upon a single prominent susceptibility gene. The Notch signalling system is one candidate system for controlling the switch and we have identified expression of Notch2 and Notch4 in seminomas and carcinoma in situ. Thus those two members of the Notch family are candidates for proto‐oncogenes that could play a role in GCT development. We have also identified a human homologue of the synaptonemal complex protein, SCP3, and have found its apparently aberrant expression in some established EC cell lines. One possibility is that abnormal regulation of such proteins involved in the synaptonemal complex could also lead to genetic instability in PGC and so also initiate tumour development.
Oyster shucking technologies: past and present
Authors: Martin Daniel E. , Hall Steven G.
Published: October 2005

A review of oyster shucking technologies from the nineteenth century to the present day is presented, comparing advantages and disadvantages of various mechanical, thermal, pressure and other technologies.
Determinants of the streptococcal surface glycoprotein GspB that facilitate export by the accessory Sec system
Authors: Bensing Barbara A. , Takamatsu Daisuke , Sullam Paul M.
Published: October 2005

GspB is a large cell‐surface glycoprotein expressed by Streptococcus gordonii M99 that mediates binding of this organism to human platelets. This adhesin is glycosylated in the cytoplasm, and is then transported to the cell surface via an accessory Sec system. To assess the structural features of GspB that are needed for export, we examined the effects of altering the carbohydrate moieties or the polypeptide backbone of GspB. Truncated, glycosylated variants of GspB were exported exclusively via the accessory Sec pathway. When glycosylation was abolished, the GspB variants were still exported by this pathway, but minor amounts could also be transported by the canonical Sec system. GspB variants with in‐frame insertions or deletions in the N‐terminus were not secreted, indicating that this domain is necessary for export. However, the N‐terminus is not sufficient for the transport of heterologous proteins, because C‐terminal fusion of passenger proteins to this domain hindered export. In contrast, fusion of GspB to a canonical signal peptide resulted in the efficient export of non‐glycosylated forms of the fusion protein via the canonical Sec pathway, whereas glycosylated forms could not be exported. Thus, the carbohydrate moieties and the atypical signal sequence of GspB interfere with export via the canonical pathway, and direct GspB towards the accessory Sec system.
Fur regulates acid resistance in Shigella flexneri via RyhB and ydeP
Authors: Oglesby Amanda G. , Murphy Erin R. , Iyer Vishwanath R. , Payne Shelley M.
Published: October 2005

Shigella flexneri requires iron for survival, and the genes for iron uptake and homeostasis are regulated by the Fur protein. Microarrays were used to identify genes regulated by Fur and to study the physiological effects of iron availability in S. flexneri. These assays showed that the expression of genes involved in iron acquisition and acid response was induced by low‐iron availability and by inactivation of fur. A fur null mutant was acid sensitive in media at pH 2.5, and acid sensitivity was also observed in the wild‐type strain grown under iron‐limiting conditions. Acid resistance of the fur mutant in minimal medium was restored by addition of glutamate during acid challenge, indicating that the glutamate‐dependent acid resistance system was not defective. Inactivation of ryhB, a small regulatory RNA whose expression is repressed by Fur, restored acid resistance in the fur mutant, while overexpressing ryhB increased acid sensitivity in the wild‐type strain. RyhB‐regulated genes were identified by microarray analysis. The expression of one of the RyhB‐repressed genes, ydeP, which encodes a putative oxidoreductase, suppressed acid sensitivity in the fur mutant. Furthermore, an S. flexneri ydeP mutant was defective for both glutamate‐independent and glutamate‐dependent acid resistance. The repression of ydeP by RyhB may be indirect, as real time polymerase chain reaction (PCR) experiments indicated that RyhB negatively regulates evgA, which encodes an activator of ydeP. These results demonstrate that the acid sensitivity defect of the S. flexneri fur mutant is due to repression of ydeP by RyhB, most likely via repression of evgA.
Characterization of a Methanosarcina acetivorans mutant unable to translate UAG as pyrrolysine
Authors: Mahapatra Anirban , Patel Asmita , Soares Jitesh A. , Larue Ross C. , Zhang Jun Kai , Metcalf William W. , Krzycki Joseph A.
Published: October 2005

The methyltransferases initiating methanogenesis from trimethylamine, dimethylamine and monomethylamine possess a novel residue, pyrrolysine. Pyrrolysine is the 22nd amino acid, because it is encoded by a single amber (UAG) codon in methylamine methyltransferase transcripts. A dedicated tRNACUA for pyrrolysine, tRNAPyl, is charged by a pyrrolysyl‐tRNA synthetase with pyrrolysine. As the first step towards the genetic analysis of UAG translation as pyrrolysine, a 761 base‐pair genomic segment in Methanosarcina acetivorans containing the pylT gene (encoding tRNAPyl) was deleted and replaced by a puromycin resistance cassette. The ΔppylT mutant lacks detectable tRNAPyl, but grows as wild‐type on methanol or acetate. Unlike wild‐type, the ΔppylT strain cannot grow on any methylamine, nor use monomethylamine as sole nitrogen source. Wild‐type cells, but not ΔppylT, have monomethylamine methyltransferase activity during growth on methanol. Immunoblot analysis indicated monomethylamine methyltransferase was absent in ΔppylT. The phenotype of ΔppylT reveals the deficiency in methylamine metabolism expected of a Methanosarcina species unable to decode UAG codons as pyrrolysine, but also that loss of pylT does not compromise growth on other substrates. These results indicate that in‐depth genetic analysis of UAG translation as pyrrolysine is feasible, as deletion of pylT is conditionally lethal depending on growth substrate.
An Extract of the Mushroom Agaricus blazei Murill Administered Orally Protects Against Systemic Streptococcus pneumoniae Infection in Mice
Authors: Bernardshaw S. , Johnson E. , Hetland G.
Published: October 2005

The aim was to investigate the antibacterial effect of the biologically active and edible mushroom Agaricus blazei Murill (AbM). A water extract of AbM or PBS control was administered orally before or with challenge to NIH/OlaHsd mice, experimentally infected intraperitoneally with the moderately virulent Streptococcus pneumoniae serotype 6B. End points were bacteraemia and survival rate. The AbM extract, protected against systemic S. pneumoniae 6B infection in the mice. It was most effective when given 24 h before inoculation but did also have protective effects when given together with challenge compared with control. The lack of antibiotic effect on pneumococci in vitro and increased levels of cytokines MIP‐2 and TNF‐α in the serum of mice receiving AbM extract, indicated that the protective effect of AbM was due to the involvement of the native immune system. This is the first report of anti‐infection effects of AbM in vivo. Our results suggest that AbM extract may be useful as additional prophylactic and possibly therapeutic treatment against bacterial and possibly other infections in humans.
Human IgG but not IgM Antibodies can Protect Mice from the Challenge with Live O6 Escherichia coli
Authors: Pontes G. N. , Massironi S. G. , Arslanian C. , Palmeira P. , Carneiro‐Sampaio M. M. S. , Nagao A. T.
Published: October 2005

We evaluated the ability of human anti‐lipopolysaccharide (LPS) O6 immunoglobulin G (IgG) and IgM antibodies to protect mice challenged with Escherichia coli serotype O6:K2ac. Purified whole IgG, commercial gammaglobulin, whole IgM‐effluent, pool of normal human serum (NHS), agammaglobulinaemic serum (test groups) or phosphate‐buffered saline (control group) was injected into adult male 18 h before a challenge with viable O6 E. coli. The mortality rate was assessed over a period of 72 h. To determine the opsonic and phagocytic activity of the antibody isotypes, we incubated peritoneal macrophages from the control and test groups collected at different times after challenge with the live bacteria with acridine orange for fluorescent analysis. Tumour necrosis factor (TNF)‐α and interleukin (IL)‐6 were quantified in serum of both the test and control groups. All mice that received commercial gammaglobulin or NHS survived. Purified whole IgG (containing 1.1 mg/l of anti‐LPS O6 IgG antibodies) protected 87.5% of the animals tested in this experiment, while whole IgM‐enriched effluent with 1.5 mg/l of anti‐LPS O6 IgM antibodies protected only 12.5%. The agamma serum showed no protective capacity compared with PBS (serving as control). The minimal concentration of anti‐LPS O6 IgG antibodies able to protect 50% of animals was 0.137 mg/l of purified whole IgG. Whole IgM‐enriched effluent showed no protective capacity independently of the concentration tested (0.048–17.0 mg/l of anti‐LPS O6 IgM antibodies). Fluorescent analysis of peritoneal macrophages from animals pretreated with purified whole IgG showed no bacteria at 8 h after the challenge. By contrast, whole IgM effluent showed an increasing number of live bacteria at the same time. Mice that had received whole IgM effluent (1.5 mg/l of anti‐LPS O6 IgM antibodies) before the challenge with LPS O6 presented 20.5 µg/l of IL‐6 and 1.5 µg/l of TNF‐α. Serum from animals pretreated with purified IgG did not present any detectable pro‐inflammatory cytokine. Our findings suggest that IgG but not IgM antibodies protect animals from a challenge with E. coli O6 serotype.
Two Swedish founder MSH6 mutations, one nonsense and one missense, conferring high cumulative risk of Lynch syndrome
Authors: Cederquist K , Emanuelsson M , Wiklund F , Golovleva I , Palmqvist R , Grönberg H
Published: October 2005

Lynch syndrome, or hereditary non‐polyposis colorectal cancer (HNPCC), is a cancer susceptibility syndrome caused by germline mutations in mismatch‐repair genes, predominantly MLH1, MSH2 and MSH6. A majority of the mutations reported are truncating, but for MSH6, missense mutations constitute over one third. Few have been proven pathogenic in functional studies or shown to segregate in families. In this study, we show segregation of the putative pathogenic MSH6 missense mutation c.1346T>C p.Leu449Pro with microsatellite instability‐high Lynch syndrome‐related tumours lacking MSH6 expression in a large 17th century pedigree. Another large family with the MSH6 nonsense c.2931C>G, p.Tyr977X mutation is similar in tumour spectra, age of onset and cumulative risk. These MSH6 families, despite their late age of onset, have a high lifetime risk of all Lynch syndrome‐related cancers, significantly higher in women (89% by age 80) than in men (69%). The gender differences are in part explained by high endometrial (70%) and ovarian (33%) cancer risks added upon the high colorectal cancer risk (60%). The several occurrences of breast cancer are not due to the MSH6 mutations. These findings are of great importance for counselling, management and surveillance of families with MSH6 mutations.
Exercise‐induced bronchoconstriction in children with atopic eczema
Authors: Caffarelli Carlo , Bacchini Pier Luigi , Gruppi Laura , Bernasconi Sergio
Published: October 2005

Non‐specific bronchial hyper‐responsiveness has been reported in most of the eczematous children even in the absence of asthmatic symptoms. We therefore investigated the occurrence of exercise‐induced bronchoconstriction (EIB) in children with atopic eczema (AE) and the predictors of EIB. Fifty‐five children referred to the paediatric clinic for AE and a control group of 17 healthy children were recruited. They all carried out a physical examination and skin prick test (SPT) both to inhalant and food allergens, prior to the exercise challenge test. Their parents filled a questionnaire on atopic diseases. They underwent exercise challenge test that consisted in free running for 6 min. Spirometric measurements were carried out before running and till 11 min after. Exercise challenge test was positive in 13 (23%) children with AE. None of the children in the control group had a positive exercise challenge test [OR (95% CI) = 1.31 (1.13–1.51); p = 0.030]. Sixteen (29%) eczematous children had a history of EIB. Such history was not reliable for identifying children who had a positive exercise test. Twenty‐nine (52%) children with AE had asthma. Allergic rhinitis affected 33 (60%) of eczematous children and allergic conjunctivitis 28 (50%). EIB was not related to any history of asthma, allergic rhinitis, allergic conjunctivitis, severity of eczema or SPT results. Our study shows that EIB is common in children with AE. Our data also indicate that screening by medical history and physical examination is not a sensitive marker of EIB. This may explain why EIB is often ignored in eczematous children.
Etiology of atopy in infancy: The KOALA Birth Cohort Study
Authors: Kummeling Ischa , Thijs Carel , Penders John , Snijders Bianca E. P. , Stelma Foekje , Reimerink Johan , Koopmans Marion , Dagnelie Pieter C. , Huber Machteld , Jansen Margje C. J. F. , De Bie Rob , Van Den Brandt Piet A.
Published: October 2005

The aim of the KOALA Birth Cohort Study in the Netherlands is to identify factors that influence the clinical expression of atopic disease with a main focus on lifestyle (e.g., anthroposophy, vaccinations, antibiotics, dietary habits, breastfeeding and breast milk composition, intestinal microflora composition, infections during the first year of life, and gene–environment interaction). The recruitment of pregnant women started in October 2000. First, participants with ‘conventional lifestyles’ (n = 2343) were retrieved from an ongoing prospective cohort study (n = 7020) on pregnancy‐related pelvic girdle pain. In addition, pregnant women (n = 491) with ‘alternative lifestyles’ with regard to child rearing practices, dietary habits (organic, vegetarian), vaccination schemes and/or use of antibiotics, were recruited through organic food shops, anthroposophic doctors and midwives, Steiner schools, and dedicated magazines. All participants were enrolled between 14 and 18 wk of gestation and completed an intake questionnaire on family history of atopy and infant care intentions. Documentation of other relevant variables started in the pregnant mother and covered the first and third trimester as well as early childhood by repeated questionnaires at 14–18, 30, and 34 wk of gestation and 3, 7, 12, and 24 months post‐partum. A subgroup of participants, including both conventional and alternative lifestyles, was asked to consent to maternal blood sampling, breast milk and a faecal sample of the infant at 1 month post‐partum, capillary blood at age 1 yr, venous blood and observation of manifestation of atopic dermatitis during home visits at the age of 2 yr (using the UK working party criteria and the severity scoring of atopic dermatitis index), and buccal swabs for DNA isolation from child–parent trios. From the start, ethical approval and informed consent procedures included gene–environment interaction studies. Follow‐up at 3 and 7 months post‐partum was completed with high response rates (respectively 90% and 88% in the conventional group, and 97% and 97% in the alternative group). The home visits at 2 yr of age will be completed in 2005. Preliminary results show that we have succeeded in recruiting a large population with various lifestyle choices with a fairly large contrast with regard to dietary habits (including organic foods, vegetarian diet), vaccination schemes and/or use of antibiotics. We have also been able to collect a large number of faecal samples (n = 1176) and capillary blood samples at age 1 yr (n = 956). Furthermore, a large proportion of the participants have consented with genetic studies. Mid 2006 we expect to report our first results on the relationship between the various exposures in early life and childhood atopy. An outline of the focus and design of the KOALA Birth Cohort Study is presented.
Selection and characterization of a nickel‐tolerant cell line from tobacco (Nicotiana tabacum cv. bright yellow‐2) suspension culture
Authors: Saito Akihiro , Higuchi Kyoko , Hirai Midori , Nakane Rie , Yoshiba Masaaki , Tadano Toshiaki
Published: October 2005

To investigate the cellular/molecular mechanisms of nickel (Ni) tolerance in high‐biomass‐producing plants, we selected a Ni‐tolerant (NIT) cell line from tobacco (Nicotiana tabacum L. cv. bright yellow‐2) suspension culture. Examination of response to various abiotic stresses showed that the NIT cells acquired remarkable tolerance to excess Ni, Cu, and Al but not to the other stresses. That the NIT cells accumulated approximately 2 mM Ni under the 700 µM Ni condition suggests that the NIT cells can hyperaccumulate Ni and possess internal detoxification mechanisms for Ni. Two different sizes of putative β‐d‐xylosidase were constitutively secreted by the NIT cells, and the NIT cells showed greater elongation than the wild‐type (WT) cells. Oxalate, citrate, 2‐oxoglutarate, and glutamate contents were constitutively two‐ or three‐fold higher in the NIT cells than in the WT cells, and histidine content was increased by up to three‐fold in the NIT cells exposed to 700 µM Ni compared to those in the WT cells. Newport green DCF diacetate (NPG), a Ni‐specific fluorescence indicator, enabled the visualization of the subcellular localization of Ni in both WT and NIT cells. The NPG fluorescence was localized mainly in the vacuoles regardless of the WT or NIT cells under excess Ni conditions. The transport of Ni‐organic acids and/or Ni‐histidine complex into the vacuoles might be an important mechanism contributing to the high Ni tolerance of and the hyperaccumulation of Ni by the NIT cells.
Mono‐carboxylic acids and their salts inhibit wound‐induced phenolic accumulation in excised lettuce (Lactuca sativa) leaf tissue
Authors: Saltveit Mikal E. , Choi Young‐Jun , Tomás‐Barberán Francisco A.
Published: October 2005

Wounding, as during excision and preparation of lettuce (Lactuca sativaL.) leaf tissue for salads, induces the synthesis and accumulation of phenolic compounds that participate in subsequent reactions that cause tissue browning. Exposure of excised 5‐mm mid‐rib segments of romaine lettuce leaf tissue to vapors of mono‐carboxylic acids or aqueous solutions of mono‐carboxylic acids or their salts inhibited wound‐induced phenolic accumulation (WIPA) and subsequent tissue browning. The decline in phenolic content followed a quadratic curve with increasing concentration, reaching a maximum inhibition after 60 min of 74 ± 8% for 50 mMsodium acetate (2 carbons, C2) and 91 ± 4% for 20 mMsodium decanoate (capric acid, C10). Respiration (i.e. carbon dioxide production) was unaffected by concentrations of formic, acetic, or propionic acids that reduced wound‐induced phenolic content or that increase ion leakage from the tissue into an isotonic mannitol solution. However, WIPA was suppressed up to 70% at concentrations (20 mMacetate) that did not increase ion leakage over that of water controls. Various acetate salts (i.e. ammonium, calcium, magnesium, and sodium) all produced the same level of inhibition. The effectiveness of the compounds increased with increasing number of carbons in the molecule from 1 to 10, and was unaffected by whether the carbons were a straight chain or branched or whether the treatment was delayed by up to 6 h. The effectiveness of butyrate (C4) in reducing WIPA (27% reduction at 20 mM) was less than that predicted from the response of the two adjacent mono‐carboxylates similarly applied: propionate (C3) (62%) and valerate (C5) (73%). It appears that, unlike the n‐alcohols, mono‐carboxylates are not interfering with the synthesis or propagation of a wound signal but are interfering with subsequent steps in the production and accumulation of wound‐induced phenolic compounds.
Immunohistochemical Identification of Amyloid, Using an Anti‐human Serum Amyloid P Component (SAP) Antibody, is Possible in Ruminants but not in Dogs and Cats
Authors: Hartig W. , Leifsson P. S. , Nielsen O. L.
Published: October 2005

Amyloidosis represents a heterogenous group of diseases that have in common the deposition of fibrils composed of proteins of beta‐pleated sheet structure, a structure which can be specifically identified by histochemistry using the Congo red or similar stains. Amyloid consists primarily of the amyloid fibrils but also of the amyloid P component (AP). This component, which is identical with the serum counterpart (SAP), is found in all types of human amyloid, and immunohistochemical identification of AP has been proposed as an adjunct to the universal, type‐independent diagnosis of human amyloidosis. In the present study of animal amyloidosis, we compared the amyloid‐specific Congo red stain with an immunohistochemical protocol using an anti‐human SAP antibody for the identification of amyloid in formalin fixed tissue samples. The species and types of amyloidoses investigated were: (i) seven cows, one yak (Bos grunniens), and one sheep affected with amyloidosis of presumed AA type, (ii) one dog with a pancreatic endocrine tumour producing amyloid of presumed AIAPP type, (iii) two cats with presumed AIAPP‐amyloidosis of the islets of Langerhans, one cat with presumed AA‐amyloidosis, and one cat with an amyloid‐producing odontogenic tumour. Intense immunostaining co‐localized with amyloid, identified by its congophilia and green birefringence, using a protocol without any antigen retrieval in each of the seven cows, the yak and the sheep. The method seemed more sensitive in the ruminants than the Congo red stain, but was unable to detect amyloid in the dog and the cats regardless of the application of various antigen retrieval protocols. However, specific identification of amyloid still rests on the Congo red method or similar histochemical techniques.
The Effect of Different Isolation Procedures on Canine Leucocyte Populations and on Lectin‐induced Lymphocyte Proliferation
Authors: De Bruin T. , De Rooster H. , Van Bree H. , Cox E.
Published: October 2005

Proliferation assays performed on peripheral blood mononuclear cells (PBMC) are commonly used in experimental and clinical immunology. A prerequisite for an in vitro assay is the ability to obtain relatively pure populations of mononuclear cells from whole blood, as contaminating polymorphonuclear cells may affect the proliferation of lymphocytes. Purification of canine leucocytes from whole blood is associated with difficulties in obtaining pure lymphocytes in high yields. The aim of this study was to optimize the lymphocyte purification from canine whole blood in terms of total cell recovery and purity, while not influencing the proliferation capacity of the isolated cells. To acquire optimal isolation of canine lymphocytes several density gradient media of different densities and osmolalities were examined. For optimal phagocyte removal, pre‐treatment of whole blood with carbonyl iron/arabic gum and/or adherence to fibrinogen pre‐coated polystyrene tissue flasks were examined. Lectin‐induced proliferation was used as measurement of cell activity of the obtained cell fractions after the different separation procedures. Canine blood pre‐treated with carbonyl iron/arabic gum followed by density gradient centrifugation with medium ‘G’ (density: 1.079 g/cm3, osmolality: 256 mOsm) and adherence to pre‐coated polystyrene tissue flask obtained the best PBMC cultures with a median lymphocyte purity of 88% and a median yield of recovered lymphocytes of 54%. This culture also resulted in the highest proliferation and subsequently the highest stimulation index upon lectin stimulation.
Effect of Insemination Volume on Uterine Contractions and Inflammatory Response and on Elimination of Semen in the Mare Uterus – Scintigraphic and Ultrasonographic Studies
Authors: Sinnemaa L. , Järvimaa T. , Lehmonen N. , Mäkelä O. , Reilas T. , Sankari S. , Katila T.
Published: October 2005

The effect of artificial insemination (AI) volume on uterine contractility and inflammation and on elimination of semen in the reproductive tract of mares was examined for 4 h after AI using two methods, scintigraphy and ultrasonography. The same doses were used in both methods: 2 and 100 ml of skim milk‐extended frozen semen. In the scintigraphic study, the number of reproductively normal mares was four per group and in the ultrasonographic study five per group. For scintigraphy, the semen was radiolabelled with technetium‐99m. The static scintigrams were acquired immediately before and 30, 60, 120, 180 and 240 min after AI. The activities in the vagina and uterus were calculated and the values for sperm that had been discharged from the mare were obtained by subtracting the counts for the uterus and vagina from the total radioactivity. The dynamic scintigrams were taken continuously for the first 30 min after AI and in 5‐min periods immediately after having acquired the static scintigrams. The uterine contractions were counted. In the ultrasonographic study, the mares were scanned before AI and at 5, 10, 15, 20, 25, 30, 60, 120, 150, 180, and 240 min after AI, for at least 1 min each time. The examinations were videotaped and contractions counted per minute. More contractions were observed with the ultrasonographic method than with the scintigraphic method. No difference was present in the number of contractions between the groups, except in the ultrasonographic study at 4 h, when the mares inseminated with 100 ml showed more contractions than did the mares inseminated with 2 ml. The intraluminal fluid was sampled with a tampon and by uterine lavage 4 h after AI in the ultrasonographic study. The numbers of polymorphonuclear leukocytes and spermatozoa were counted, but the differences between the groups were not significant. Under our experimental conditions and with the number of mares examined, the volume of the AI dose had an insignificant effect on contractility – with the exception at 4 h – and inflammatory reaction and on semen elimination in the uterus.
Influence of Partus Induction on the pH Value in the Blood of Newborn Piglets
Authors: Wehrend A. , Stratmann N. , Failing K. , Bostedt H.
Published: October 2005

Partus induction is an important integrative component of reproduction management in the swine industry. In this context, the question is to what extent the application of uterotonic substances may influence the vitality of piglets. The blood pH value is used as a laboratory objective diagnostic parameter of neonatal acidosis. This parameter of the acid–base balance was examined in piglets born spontaneously at the 115th to 116th day of gestation (group 1, n = 57) and after two different partus induction protocols starting at the 114th day of gestation (group 2: combination of 175 μg of cloprostenol and 20 IU of oxytocin, n = 55; group 3: 175 μg of cloprostenol and 35 μg of carbetocin, n = 56). Initial blood samples were taken from each piglet at birth and 2 h later. The results show that the different protocols have no negative influence on the initial pH value and the compensation of neonatal acidosis until the second hour post‐natum.
Immunohistochemical Observations on TSH Secreting Cells in Pituitary Glands of Goat Kids with Congenital Goitre
Authors: Ozmen O. , Haligur M.
Published: October 2005

Pituitary glands of normal‐termed stillborn goat kids with congenital goitre and normal‐termed stillborn goat kids without congenital goitre were examined macroscopically, histopathologically and immunohistochemically. Thyroid glands of these animals were also examined grossly and microscopically. The pituitary glands of kids with goitre were larger than those of normal kids, and on histopathological examination there was hyperplasia of the acidophil cells in the ventral part of the glands. However, it was impossible to distinguish thyroid stimulating hormone (TSH)‐secreting cells from other acidophil cells in sections stained with haematoxylin and eosin (HE). Red granules were observed in the cytoplasm of these hyperplastic cells in periodic acid‐Schiff (PAS)‐stained sections. Sections were also immunostained with an antibody against TSH using the streptavidin–biotin peroxidase technique. Immunohistochemistry revealed TSH‐secreting cells to have increased in number in the pituitary glands of kids with congenital goitre because of the extensive proliferation when compared with those of normal kids. The present study indicated that the presence of multiple fetuses (twins or triplets) may be a predisposing factor for congenital goitre.
Enhanced Non‐esterified Fatty Acids and Corticosterone in Blood Plasma of Chickens Treated with Insulin are Significantly Depleted by Reverse T3: Minor Changes in Hypoglycaemia
Authors: Niezgoda J. , Bobek S. , Wrońska‐Fortuna D.
Published: October 2005

Previously, it has been observed that dexamethasone or adrenaline‐induced hyperlipaemia in blood of chicken was significantly reduced after administration of reverse triiodothyronine (rT3). The present experiment was performed on chicken to determine the altered circulating non‐esterified fatty acids (NEFA) induced by physiologically enhanced endogenous corticosterone and catecholamines may also be influenced by rT3. Rise of both hormones were induced by insulin administration. Changes in circulating glucose, corticosterone and catecholamines were additionally measured. Following insulin injection blood glucose fell on the average by 32.7% below control at 2 h of the experiment. Additional treatment with rT3(rT3 + insulin group) gradually attenuated this decrease and at 4 and 6 h of the experiment it was 17.1% and 12.9% below control, respectively, suggesting on slight inhibition by rT3of insulin‐stimulated glucose utilization. Exposure to insulin significantly increased NEFA levels to about 670% above control group. Additional treatment with rT3reduced this increase to 309% of control, suggesting inhibition of lipolysis by rT3. Similar alterations were observed in plasma corticosterone levels. Insulin treatment peaked the corticosterone levels maximally by 507.6% above control. Additional treatment with rT3abolished this rise in the averages to 194.2% above control, possibly by interaction of rT3with hypothalamo‐adrenal axis. Insulin injection increased plasma catecholamines on the average by 21.5% and 53.4% for adrenaline and noradrenaline respectively. Supplementary treatment with rT3intensified this rise by 55.6% and 71.6% respectively. The obtained results suggest on inhibitory effect of rT3on hypoglycaemia, hyperlipaemia and plasma corticosterone concentrations in chickens treated with insulin. Contrary to this, rT3enhanced the rise of plasma catecholamines due to insulin treatment. The obtained data favour the assumption that hypometabolic properties of rT3depends mainly upon reduced supply of NEFA as a result of restricted lipolysis and to a lesser extent upon the supply of glucose.
Osseous Lesion of the Calcaneus Following the Use of Shock Wave Therapy in a Horse
Authors: Diakakis N. , Patsikas M. N. , Dessiris A. K.
Published: October 2005

An 8‐year‐old Dutch warmblood gelding was presented with a mechanical lameness (2/10) because of the presence of a soft tissue injury on the top of the right tuber calcanei. Plain radiographs of the tarsus demonstrated the presence of soft tissue swelling caudal to the right tuber calcanei, without osseous involvement, and ultrasonography revealed excessive scar tissue within and around the superficial digital flexor tendon. Extra‐corporeal shock wave therapy was applied on the right hock to decrease the amount of scar tissue. One month after the therapy the lameness was greater (3/10) and a marginal increase in the size of the swelling was found. Periosteal new bone formation associated with an ill‐defined radiolucent area and two bony fragments were detected radiographically at the caudo‐proximal aspect of the right tuber calcanei. A blister containing oil of croton, camphor, pine and thyme, turpentine and cantharides was applied on the right calcaneus. Twenty days after blister application, the size of the swelling had been reduced by 50% and the degree of lameness had also been decreased (1/10). On clinical re‐evaluation 6 months after treatment, the degree of lameness was stable (1/10) and flexion test of the limb was negative.
Microsporum gypseumisolated from a feline case of dermatophytosis
Authors: Kano R. , Yasuda K. , Nakamura Y. , Hasegawa A.
Published: October 2005

Summary.The 1‐ to 2‐month‐old female cross‐breed cat presented with alopecia, erythema and many crusts were present on the tail. Microscopic examination of crusts from the tail disclosed epithelial debris, exudate, mycelium, and arthrospores.Microsporum gypseumwhich is rarely isolated from cats as a causative agent of dermatophytsis was cultured from the crusts on a 1/10 Sabouraud glucose agar at 27°C for 1 week. The isolate ofM. gypseumfrom the cat was examined by random amplification of polymorphic DNA (RAPD), chitin synthase 1 gene (CHS1) sequence and mating experiments. The RAPD band patterns of the clinical isolate ofM. gypseumwas identical to those of tester strains ofArthroderma gypseum. Nucleotide sequence analysis of the CHS1 gene fragments from the isolate and a tester strain ofA. gypseumshowed 100% similarity. The mating experiments on the clinical isolate ofM. gypseumcompletely agreed with the results from RAPD and CHS1 gene sequence. The isolate from the cat was confirmed to beA. gypseum(–) mating type, which was consistent with the result of mycological examination by molecular analyses.
Case Report. Successfully treated subcutaneous infection with Sporothrix schenckii in Turkey
Authors: Koç A. N. , Ukşal Ü. , Oymak O.
Published: October 2005

Summary. Sporotrichosis is rare in Turkey. We report a 48‐year‐old man who had subcutaneous sporotrichosis caused by Sporothrix schenckii that was successfully treated with short‐term itraconazole and potassium iodide. The isolate was susceptible to itraconazole also in vitro. Short‐term itraconazole and potassium iodide should be agents of choice for treatment of subcutaneous sporotrichosis. However, treatment is controversial both in choice of agent used and in duration of therapy.
Case Report. Disseminated tinea of the verrucous type due to Epidermophyton floccosum
Authors: Qiangqiang Z. , Limo Q. , Qixian Q.
Published: October 2005

Summary. A case is presented of an 8‐year‐old boy suffering from disseminated verrucous lesions with some purulent drainage on his right foot, shank, thigh, scrotum, penis, perianal area and abdomen. Epidermophyton floccosum was isolated from these lesions. The patient was treated with oral ketoconazole for 4 months and the lesions resolved completely. There was no evidence of relapse during a 12‐month follow‐up.
Oral colonization by Candida spp. among AIDS household contacts
Authors: Milan Eveline P. , Kallás E. G. , Costa P. R. de O. , Da Matta D. A. , Lopes Colombo A.
Published: October 2005

Summary. This study was designed to investigate the oral yeast colonization rate of household contacts of AIDS patients. Sixty‐four AIDS household contacts were sequentially enrolled along with 103 HIV‐negative blood bank donors (control group). Samples were obtained by swabbing the oral mucosa. Yeast isolates were identified by classical methods and antifungal susceptibility testing was performed according to NCCLS microbroth assay. Candida spp. was recovered from the oral cavity of 33% of the AIDS household contacts, in contrast with 14% of the control group (P = 0.003 or P = 0.04 after adjusting for oral prosthesis use). Candida albicans was the most frequently isolated species in both groups. All of the isolates were susceptible to fluconazole, itraconazole and ketoconazole. In conclusion, we were able to demonstrate a higher colonization rate in the AIDS household contacts group compared with the control group. No resistant isolates to antifungal drugs was observed. We suggest that the contact with AIDS patients may play a role as a risk factor for developing oral colonization by Candida spp.
Identification of Sporothrix schenckii based on sequences of the chitin synthase 1 gene
Authors: Kano R. , Nakamura Y. , Watanabe S. , Tsujimoto H. , Hasegawa A.
Published: October 2005

Summary. Sporothrix schenckii is pathogenic to human and animals. To detect S. schenckii in the tissue, we designed specific oligonucleotide primers based on the chitin synthase 1 gene. Amplification products were selectively obtained from S. schenckii DNA. Polymerase chain reaction analysis with the primer pair S2–R2 was able to detect 10 pg genomic DNA of S. schenckii with ethidium bromide staining. This detection system will be useful as a microbiological tool for the diagnosis of human and animal sporotrichosis.
Mucormycoses
Authors: Eucker J. , Sezer O. , Graf Barbara , Possinger K.
Published: October 2005

Summary. Over recent years the clinical importance of mucormycosis has significantly increased. Most frequently mucormycosis occurs in neutropenic patients with haematological diseases. It is caused by fungi of the order Mucorales. The clinical patterns of the disease produced by different genera or species of Mucorales are virtually identical. Rhizopus, Absidia, Rhizomucor and Mucor are the organisms most commonly isolated from patients who suffer from mucormycosis. Diagnosis of mucormycosis is difficult as it is based on culture methods or microscopy of clinical specimens. The diagnosis is often only made after a delay or even post‐mortem. Therapy includes surgical intervention if possible and is based on systemic amphotericin B (conventional or liposomal).
Case Report. Lymphatic vessel‐type sporotrichosis: immunohistochemical evaluation and cytokine expression pattern
Authors: Tanuma H. , Asai T. , Abe M. , Nishiyama S. , Katsuoka K.
Published: October 2005

Summary. A 57‐year‐old male carpenter living in Sagamihara, Kanagawa Prefecture, visited the Department of Dermatology of Kitasato University Hospital because an ulcer which appeared in his left forearm around May 1992 had spread gradually. An oval, shallow ulcer measuring 39 mm × 18 mm was found on the flexor aspect of the left forearm. Histopathological examination showed partial ulceration on the epidermis and marked cell infiltration throughout the entire dermal layer with an abscess in the centre and granulomatous reactions around it. PAS‐positive spores were present between infiltrating cells and in giant cells in abscess and in granulomatous reactions. The skin lesion rapidly disappeared after beginning treatment with 125 mg day−1 terbinafine and only a slight redness remained 14 weeks after starting the treatment. At this time the culture was negative. We conducted immunohistochemical examinations of the affected skin before, during and after starting treatment with terbinafine and studied local expression of cytokines at the affected lesion.
Larval head anatomy ofHeterogynis penella(Zygaenoidea, Heterogynidae), and a general discussion of caterpillar head structure (Insecta, Lepidoptera)
Authors: Vegliante F.
Published: October 2005

The internal head anatomy (and the peculiar integumental structure of the epicranial notch region) ofHeterogynis penellalarvae are described; special attention is paid to the skeleto‐muscular and nervous systems and to the cephalic glands. Transverse ligaments connect the apodemes of the mandibular adductor muscles of both sides and the anterior maxillo‐labial articulations of both sides. The two ligaments are linked to each other by a thin, apparently acellular membrane. An accessory, trilobed mandibular gland is present. A putative stretch receptor, connecting the oblique dorsal cibarial dilators of both sides, is described for the first time in a lepidopterous larva and its importance in assessing the homology of these muscles is discussed. The presence of cibarial sensilla, previously predicted in other caterpillars on the basis of behavioural experiments and observations of the nerve pattern, is confirmed. The structural diversity of larval head anatomy in ditrysian Lepidoptera is discussed, with particular emphasis on the innervation of the corpora cardiaca and corpora allata and of the sensilla of the head capsule.
Breeding success of the colonist species Bubulcus ibis (Linnaeus, 1758) and four native species
Authors: Petry Maria Virginia , Fonseca Vanda Simone Da Silva
Published: October 2005

This work evaluated the breeding success of Bubulcus ibis (Ardeidae), Egretta thula, Nycticorax nycticorax, Phimosus infuscatus and Plegadis chihi in Rio Grande do Sul during two consecutive nesting seasons. Mean clutch size for B. ibis was 2.59 in 1998/1999 and 2.49 in 1999/2000. The species with both the smallest and largest clutch was P. infuscatus, with 2.04 in 1999/2000, and 3.16 in 1998/1999. The highest mean number of nestlings per nest was 2.86 for P. chihi in 1998/1999 and the lowest was 1.85 for B. ibis in 1998/1999 and P. infuscatus in 1999/2000. In both nesting seasons all the species had major success, laying three eggs and raising two nestlings, exept for B. ibis. The breeding success for P. chihi varied from 1.54 young/nest in 1998/1999 to 0.4 in 1999/2000, whereas B. ibis ranged from 1.16 in the first period to 0.99 in the second.