bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of Maspin in Mammary Gland Tumors of the Dog
Authors: de los Monteros A. Espinosa , Millán M. Y. , Ramírez G. A. , Ordás J. , Reymundo C. , de las Mulas J. Martín
Published: May 2005

Maspin is a serine protease inhibitor that inhibits tumor invasion and metastasis in human breast cancer and is consistently expressed by mammary myoepithelial cells (MECs). To analyze the value of maspin as a marker of the MEC layer of the normal and tumoral canine mammary gland, the immunohistochemical expression of maspin was studied in formalin-fixed tissues from 55 benign and malignant tumors (40 tumors also contained the surrounding normal mammary gland) using a commercially available monoclonal antibody. Periacinar and periductal MECs of all 40 normal mammary glands were stained by the anti-human maspin monoclonal antibody, and immunoreactivity was observed in the nucleus and cytoplasm of these cells. In addition, maspin was found in 53 (98%) of the tumors studied, reacting with the MECs in 100% of benign tumors and 93% of malignant tumors and to the epithelial cells of 16% of benign and 73% of malignant tumors. In the MEC compartment, immunoreactivity was observed in the cytoplasm of hypertrophic MECs, fusiform MECs, stellate MECs, rounded (myoepithelial) cells, and chondroblasts. In the epithelial cell compartment, immunoreactivity was observed in the cytoplasm of cells with and without squamous differentiation. Stromal myofibroblasts were unreactive. Maspin appears to be a very sensitive marker of the normal and neoplastic myoepithelium that, contrary to smooth muscle differentiation markers, does not stain stromal myofibroblasts. In addition, a subset of neoplastic epithelial cells reacted with the maspin antibody. The relationship between maspin expression in different cellular compartments of canine mammary carcinomas and the biologic aggressiveness of the disease remains to be elucidated.
Acute and Chronic Gas Bubble Lesions in Cetaceans Stranded in the United Kingdom
Authors: Jepson P. D. , Deaville R. , Patterson I. A. P. , Pocknell A. M. , Ross H. M. , Baker J. R. , Howie F. E. , Reid R. J. , Colloff A. , Cunningham A. A.
Published: May 2005

The first evidence suggestive of in vivo gas bubble formation in cetacea, including eight animals stranded in the UK, has recently been reported. This article presents the pathologic findings from these eight UK-stranded cetaceans and two additional UK-stranded cetacean cases in detail. Hepatic gas-filled cavitary lesions (0.2-6.0 cm diameter) involving approximately 5–90% of the liver volume were found in four (two juvenile, two adult) Risso's dolphins ( Grampus griseus), three (two adult, one juvenile) common dolphins ( Delphinus delphis), an adult Blainville's beaked whale ( Mesoplodon densirostris), and an adult harbour porpoise ( Phocoena phocoena). Histopathologic examination of the seven dolphin cases with gross liver cavities revealed variable degrees of pericavitary fibrosis, microscopic, intrahepatic, spherical, nonstaining cavities (typically 50–750 μm in diameter) consistent with gas emboli within distended portal vessels and sinusoids and associated with hepatic tissue compression, hemorrhages, fibrin/organizing thrombi, and foci of acute hepatocellular necrosis. Two common dolphins also had multiple and bilateral gross renal cavities (2.0–9.0 mm diameter) that, microscopically, were consistent with acute ( n = 2) and chronic ( n = 1) arterial gas emboli-induced renal infarcts. Microscopic, bubblelike cavities were also found in mesenteric lymph node ( n = 4), adrenal ( n = 2), spleen ( n = 2), pulmonary associated lymph node ( n = 1), posterior cervical lymph node ( n = 1), and thyroid ( n = 1). No bacterial organisms were isolated from five of six cavitated livers and one of one cavitated kidneys. The etiology and pathogenesis of these lesions are not known, although a decompression-related mechanism involving embolism of intestinal gas or de novo gas bubble (emboli) development derived from tissues supersaturated with nitrogen is suspected.
Epithelial-Stromal Tumor of the Seminal Vesicles in the Transgenic Adenocarcinoma Mouse Prostate Model
Authors: Tani Y. , Suttie A. , Flake G. P. , Nyska A. , Maronpot R. R.
Published: May 2005

The transgenic adenocarcinoma mouse prostate (TRAMP) model, designed for researching human prostatic cancer, was genetically engineered to harbor a transgene composed of the simian virus 40 Large-T/small-t antigen promoted by the rat probasin gene. In addition to prostatic neoplasms, the TRAMP mouse develops tumors in the seminal vesicles. This study was conducted to evaluate the pathology and histogenesis of TRAMP seminal vesicle neoplasms. Tissues of accessory sex organs harvested from 72 TRAMP mice of various ages (11-40 weeks of age) were fixed in neutral buffered formalin and stained with hematoxylin and eosin, desmin, 5-bromo-2′-deoxyuridine (BrdU, treated animals only), and SV40 Large-T antigen (SV40-Tag). In the seminal vesicles, we found neoplastic stromal cells that emerged multicentrically just beneath the epithelium, densely packed between the epithelium and the smooth muscle layer These stromal cells frequently exhibited mitotic figures and showed BrdU incorporation and SV40-Tag protein expression in the nuclei and immunopositivity for desmin. The proliferative mesenchymal cells were lined by cuboidal to columnar epithelium. Some of the larger papillary, polypoid lesions exhibited a phyllodes pattern resembling that seen in mixed epithelial-stromal tumors of the breast, prostate, and seminal vesicles of humans. Although the epithelium was negative for SV40-Tag and showed only occasional incorporation of BrdU, it clearly participated in the biphasic proliferation, forming papillary, cystic, and tubuloglandular structures. No conclusive evidence of malignancy (invasion or metastasis) was identified. Our recommended diagnosis of this lesion in the seminal vesicles is epithelial-stromal tumor.
Malignant Seminoma with Metastasis, Sertoli Cell Tumor, and Pheochromocytoma in a Spotted Dolphin (Stenella frontalis) and Malignant Seminoma with Metastasis in a Bottlenose Dolphin (Tursiops truncatus)
Authors: Estep J. S. , Baumgartner R. E. , Townsend F. , Pabst D. A. , Mclellan W. A. , Friedlaender A. , Dunn D. G. , Lipscomb T. P.
Published: May 2005

Seminoma with metastasis was diagnosed in a spotted dolphin ( Stenella frontalis) and an Atlantic bottlenose dolphin ( Tursiops truncatus). Sertoli cell tumor and pheochromocytoma were also diagnosed in the spotted dolphin. The spotted and bottlenose dolphins were adult males that stranded and died on the coasts of northwest Florida and southeast North carolina, respectively. Neoplasia is infrequently reported in cetaceans. This is the first report of seminoma, Sertoli cell tumor, and pheochromocytoma in a dolphin, the first report of three distinct neoplasms in a dolphin, and one of the few reports of malignant neoplasia in dolphins.
Low-grade Glial Tumor with Features of Astroblastoma in a Dog
Authors: Cowart J. R. , Schulman F. Y. , Mena H.
Published: May 2005

A 12-year-old, neutered, male Belgian Malinois/Great Dane cross dog presented with a 5-month history of weakness and lack of endurance followed by acute onset of rear limb ataxia. At autopsy, a 9 x 16 mm, multilobular, firm, white to tan, expansile mass was found in the cerebellum. Mild dilatation of the lateral ventricles was also noted. Histologically, there was a well-demarcated glial neoplasm composed of medium-sized astrocytic elements that had homogeneous cytoplasm, sometimes with globular eosinophilic inclusions, irregular peripherally located nuclei with a single nucleolus, and short cytoplasmic processes. Prominent peri-vascular pseudorosettes with cellular processes in contact with blood vessels were present. Some blood vessels exhibited hyalinized walls. Mitotic figures were not observed. Immunohistochemically, neoplastic cells expressed glial fibrillary acidic protein and vimentin. These features are consistent with an astroblastoma. This is the first clinicopathologic correlation and detailed description of a low-grade glial tumor with features of astroblastoma in a dog.
Cutaneous Angiomatosis in a Young Dog
Authors: Kim Y. , Reinecke S. , Malarkey D. E.
Published: May 2005

A 1-year-old, spayed, female, mixed-breed dog had two reddish-purple cutaneous lesions, one on the right dorsal antebrachium and the other on the right shoulder. The lesions consisted of approximately 13 x 3 cm and 15 x 10 cm, irregular, patchy regions of 0.5-3.0 cm, circular, sometimes raised, reddish-purple swellings resembling ecchymoses. The lesion on the antebrachium had been noticed since the dog was adopted at 6 months of age and appeared to have increased in size over an 11-week period, at which time skin punch biopsy revealed an infiltrative pattern of well-differentiated blood vessels leading to an interpretation that the lesion was a well-differentiated hemangiosarcoma. The second lesion was revealed when the dog had its fur shaved in that area during surgical preparation to excise the antebrachial lesion. No other skin lesions were found on the dog. Microscopically, there was a widely disseminated and infiltrative-like pattern of benign-appearing small blood vessels, which were throughout the superficial and deep dermis and subcutis. Although the disseminated nature suggested malignancy, the histologic appearance of well-differentiated small blood vessels and nonprogressive clinical features indicate that the lesions were benign. The dog has been followed for 6 years and to date has no evidence of progression of the antebrachial lesion or shoulder lesion. To the authors' knowledge, this is the first report of a congenital angiomatosis-like lesion in a young dog, with extensive involvement of the forelimb.
Gastritis-associated Adenocarcinoma and Intestinal Metaplasia in a Syrian Hamster Naturally Infected with Helicobacter Species
Authors: Nambiar P. R. , Kirchain S. , Fox J. G.
Published: May 2005

The objective of this study was to evaluate stomachs of 2-year-old Syrian hamsters that were naturally colonized by multiple Helicobacter species including Helicobacter aurati. A previous report on 7- to 12-month-old Syrian hamsters described chronic gastritis and intestinal metaplasia, a putative preneoplastic lesion in the stomach, without cancer. This report describes an invasive adenocarcinoma at the pyloric-duodenal junction in one of nine hamsters at a site of helicobacter-associated inflammation and marked intestinal metaplasia. Ceca of nine of nine animals were culture positive and polymerase chain reaction positive for Helicobacter spp. Furthermore, immunohistochemical analysis of the stomach using a H. pylori polyclonal antibody detected positive-staining bacteria within the pyloric region of three of nine hamsters including the neoplastic glands. However, argyrophilic bacteria were demonstrated only within the stomach of the hamster with gastric adenocarcinoma. This is a first report of gastric adenocarcinoma in helicobacter-infected hamsters. Syrian hamsters appear suitable as potential model for studying development of helicobacter-associated gastric adenocarcinomas.
Intestinal Stromal Tumors in a Simian Immunodeficiency Virus-infected, Simian Retrovirus-2 Negative Rhesus Macaque (Macaca mulatta)
Authors: Bielefeldt-Ohmann H. , Barouch D. H. , Bakke A. M. , Bruce A. G. , Durning M. , Grant R. , Letvin N. L. , Ryan J. T. , Schmidt A. , Thouless M. E. , Rose T. M.
Published: May 2005

Multifocal submucosal stromal tumors were diagnosed in a 5.5-year-old rhesus macaque ( Macaca mulatta) experimentally infected with simian immunodeficiency virus, strain SIVsmE660, and CD4+ T cell depleted. The animal was negative for simian retroviruses, SRV-1, -2, and -5. Polymerase chain reaction analysis of DNA from tumor and spleen tissue revealed abundant, preferential presence of retroperitoneal fibromatosis herpesvirus, the macaque homologue of the Kaposi sarcoma-associated herpesvirus (human herpesvirus-8), in the tumors. This was corroborated by demonstration of viral latent nuclear antigen-1 in the nuclei of a majority of the spindeloid tumor cells. Low levels of an additional macaque herpesvirus, rhesus rhadinovirus, were also detected in the spleen and tumor tissues. The spindeloid cells labeled positively for vimentin and CD117 but were negative for CD31, CD68, desmin, and smooth muscle cell actin. Collectively, these findings suggest a relation to but not absolute identity with simian mesenchymoproliferative disorders (MPD) or typical gastrointestinal stromal tumors (GISTs).
Radiochemical separation of actinides from environmental samples for determination with DF-ICP-MS and alpha spectrometry
Authors: Ayranov Marin , Krähenbühl Urs , Sahli Hans , Röllin Stefan , Burger Mario
Published: May 2005

The purpose of this study was to develop an optimised sample preparation method for the determination of Pu, Am, and Cm in environmental samples, soils, and sediments, and to compare the sensitivities of DF-ICP-MS, alpha spectrometry, and liquid scintillation alpha spectrometry. Borate fusion was used for sample material digestion. Two methods based on TOPO liquid–liquid extraction and combination of TEVA and TRU extraction chromatography showed highest yields of 92.7% for Pu, 86.8% for Am, and 87.6% for Cm. The detection limits were between 0.014–0.034 Bq/kg for 239Pu, 0.014–0.093 Bq/kg for 240Pu, and 0.011–1.20 Bq/kg for 241Am. Analytical results for 238Pu, 239Pu, 240Pu, 241Pu and 241Am and their isotope ratios in certified reference materials (IAEA-135 and IAEA-300) indicated very good accuracy of the methods proposed.
Microscale characterization of uranium(VI) silicate solids and associated neptunium(V)
Authors: Douglas Matthew , Clark Sue B. , Friese Judah I. , Arey Bruce W. , Buck Edgar C. , Hanson Brady D. , Utsunomiya Satoshi , Ewing Rodney C.
Published: May 2005

The uranium(VI) silicate phases uranophane, Ca[(UO2)(SiO3OH)]2·5H2O, and sodium boltwoodite, Na[(UO2)(SiO3OH)]·1.5H2O, were synthesized in the presence of small, variable quantities (0.5–2.0 mol % relative to U) of pentavalent neptunium (Np(V), as NpO2
+), to investigate the nature of its association with these U(VI) solid phases. Solids were characterized by X-ray powder diffraction (XRD), gamma spectrometry (GS), scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDS), and transmission electron microscopy (TEM) with electron energy-loss spectroscopy (EELS). Neptunium concentration was determined in the bulk solid phases by GS and was found to range from 780–15800 μg/g. In some cases, Np distributions between the aqueous and solid phases were monitored, and 78–97% of the initial Np was associated with the isolated solid. Characterization of individual crystallites by TEM/EELS suggests the Np is associated with the U(VI) phase. No discrete Np phases, such as Np oxides, were observed. Because the U(VI) silicates are believed to be important solubility-controlling solids on a geologic timescale, these results suggest that the partitioning of the minor actinides to these solids must be considered when assessing the performance of a waste repository for spent nuclear fuel.
Crystal structure of new uranyl oxalates (NH4)2[UO2(C2O4)2· H2O]·2H2O and (NH4)2-x(N2H5)x[(UO2)2(C2O4)3]·3H2O (x=0 and x=1). Comparison with other uranyl oxalates
Authors: Chapelet-Arab B. , Nowogrocki G. , Abraham F. , Grandjean S.
Published: May 2005

Two new ammonium uranyl oxalates (NH4)2-[UO2(C2O4)2H2O]·2H2O (1) and (NH4)2[(UO2)2(C2O4)3]· 3H2O (2) and one mixed ammonium hydrazinium uranyl oxalate (NH4)(N2H5)[(UO2)2(C2O4)3]·3H2O (3) have been prepared. The crystal structures of the three compounds have been determined by single-crystal X-ray diffraction. The crystal structures were solved by the Patterson method and Fourier difference techniques, and refined by a least square method on the basis of F2for all unique reflections. Crystallographic data: 1, hexagonal, space group P6122,a=8.143(2) Å,c=36.179(18) Å,Z=6,R1= 0.0268 for 105 parameters with 1223 reflections withI≥ 2σ(I); 2, monoclinic, space groupP21/n,a=8.705(3) Å,b= 19.454(6) Å,c=5.5105(18) Å, β=101.741(5)°,Z=2,R1= 0.0314 for 139 parameters with 2059 reflections withI≥ 2σ(I); 3, monoclinic, space groupP21/n,a=8.958(3) Å,b=19.601(7) Å,c= 11.007(4) Å, β=112.928(5)°,Z=4,R1=0.0363 for 131 parameters with 3346 reflections withI≥ 2σ(I). The crystal structure of 1 contains [UO2(C2O4)2· H2O]2−anions, NH4+cations and water molecules. In the anion, the linear uranyl group is coordinated by two bidentate oxalate anions and one water molecule giving a pentagonal bipyramidal environment for the uranium atom. The structures of 2 and 3 consist of two-dimensional networks built from uranyl and oxalate ions forming hexagonal rings. The oxalate ions are tetradentate, each bridging two uranyl ions which are coordinated by three oxalate groups to give distorted hexagonal bipyramidal environments for the uranium atoms. In 3, the ammonium and hydrazinium ions are ordered in successive layers. The various possible coordinations for the uranyl cation and the great versatility of the oxalate anion are magnificently illustrated in the monovalent ion-uranyl double oxalates series and other structures are expected by the diversification of the synthesis methods and experimental conditions.
Extraction of U(VI) with unsymmetrical N-methyl-N-decylalkylamide in toluene
Authors: Cui Yu , Sun Guoxin , Zhang Zhenwei , Li Yexin , Sun Sixiu
Published: May 2005

A number of structurally related new unsymmetrical monoamides with different substituting groups, N-methyl-N-decyloctylamide (MDOA), N-methyl-N-decyldecanamide (MDDA), and N-methyl-N-decyldodecanamide (MDDOA), were synthesized. The extraction of U(VI) from nitric acid medium by the synthesized monoamides diluted with toluene was investigated. The effect of nitric acid concentration, sodium nitrate concentration, extractant concentration and temperature on the distribution ratio was studied. The stoichiometry of the main extracted species confirms to UO2(NO3)2·2A (A, amidic extractant). The values of the thermodynamic parameters were determined and the extraction reactions were found exothermic. The IR spectral study was also made of the extracted species.
Speciation of Tc(IV) in chloride solutions by capillary electrophoresis
Authors: Liu Xiaolan , Poineau Frédéric , Fattahi Massoud , Grambow Bernd , Vichot L.
Published: May 2005

A method for speciation of Tc(IV) species (TcCl6
2− and TcCl5(H2O)−) in chloride solutions, using capillary electrophoresis (CE) technique was developed. The proposed method has overcome the difficulties of unstable oxidation states analysis by shortening their travel time in the capillary. TcCl6
2− and TcCl5(H2O)− were thus separated without being hydrolyzed and polymerized, and their UV/Vis spectra were recorded. With a 1 M HCl/NaCl buffer solution (pH=1), the electrophoretic mobilities were determined as 5.47×10−4 cm2/Vs for TcCl6
2− and 2.13×10−4 cm2/Vs for TcCl5(H2O)− at 25 °C. The total analysis time for one run is 12 minutes.
Seasonality of growth response to GH therapy in prepubertal children with idiopathic growth hormone deficiency
Authors: Land C , Blum W F , Stabrey A , Schoenau E
Published: May 2005

Objective: Longitudinal growth of children exhibits seasonal variation. In both healthy children and in children with growth hormone (GH) deficiency (GHD) receiving GH therapy, growth rate is maximal during spring and summer. In the present study, we analyzed the growth response to GH therapy in children with GHD as a function of the season when therapy was started.

Subjects and methods: Anthropometric measurements and biochemical analyses of GH secretion status and bone formation were longitudinally assessed in a cohort of 52 prepubertal children with GHD (14 girls, mean age 7.6 years) who were treated with a fixed dose of GH (0.025 mg/kg/day).

Results: Auxological assessments over the 2-year observation period revealed a significantly better growth response to GH therapy in children who started therapy between the spring and summer (group 1) compared with children who started in the autumn or winter (group 2). The difference was largest in the initial 3-month treatment period (35%; P < 0.01). The initial better gain in height of group 1 was sustained during the study period. Baseline peak GH levels during stimulation tests and insuin-like growth factor-I levels did not differ between the two groups. However, group 1 had significantly higher bone resorption and formation markers, either at the start or shortly after initiation of GH treatment. This suggests that children with GHD have higher bone turnover during spring and early summer, irrespective of GH therapy.

Conclusions: In summary, this study suggests that the season of GH initiation is a determinant of the initial growth response to GH replacement in prepubertal children with GHD.
Thyrotrophin-releasing hormone receptor 1 and prothyrotrophin-releasing hormone mRNA expression in the central nervous system are regulated by suckling in lactating rats
Authors: Fjeldheim Åse-Karine , Høvring Per Ivar , Løseth Ole-Petter , Johansen Per Wiik , Glover Joel C , Matre Vilborg , Olstad Ole Kristoffer , Reppe Sjur , Gordeladze Jan O , Walaas S Ivar , Gautvik Kaare M
Published: May 2005

Background: The accepted function of the hypothalamic peptide, thyrotrophin-releasing hormone (TRH), is to initiate release of thyrotrophin (TSH) from the pituitary. A physiological role for TRH in lactating rats has not yet been established.

Methods: Tissues were prepared from random-cycling and lactating rats and analysed using Northern blot, real time RT-PCR and quantitativein situhybridisation.

Results: This study demonstrates that TRH receptor 1 (TRHR1) mRNA expression is up-regulated in the pituitary and in discrete nuclei of the hypothalamus in lactating rats, while proTRH mRNA expression levels are increased only in the hypothalamus. The results were corroborated by quantitativein situanalysis of proTRH and TRHR1. Bromocriptine, which reduced prolactin (PRL) concentrations in plasma of lactating and nursing rats, also counteracted the suckling-induced increase in TRHR1 mRNA expression in the hypothalamus, but had an opposite effect in the pituitary. These changes were confined to the hypothalamus and the amygdala in the brain.

Conclusions: The present study shows that the mechanisms of suckling-induced lactation involve region-specific regulation of TRHR1 and proTRH mRNAs in the central nervous system notably at the hypothalamic level. The results demonstrate that continued suckling is critical to maintain plasma prolactin (PRL) levels as well as proTRH and TRHR1 mRNA expression in the hypothalamus. Increased plasma PRL levels may have a positive modulatory role on the proTRH/TRHR1 system during suckling.
Effect of long-term continuous methimazole treatment of hyperthyroidism: comparison with radioiodine
Authors: Azizi F , Ataie L , Hedayati M , Mehrabi Y , Sheikholeslami F
Published: May 2005

Objective: To investigate the long-term effects of continuous methimazole (MMI) therapy.

Design and methods: Five hundred and four patients over 40 years of age with diffuse toxic goiter were treated with MMI for 18 months. Within one year after discontinuation of MMI, hyperthyroidism recurred in 104 patients. They were randomized into 2 groups for continuous antithyroid and radioiodine treatment. Numbers of occurrences of thyroid dysfunction and total costs of management were assessed during 10 years of follow-up. At the end of the study, 26 patients were still on continuous MMI (group 1), and of 41 radioiodine-treated patients (group 2), 16 were euthyroid and 25 became hypothyroid. Serum thyroid and lipid profiles, bone mineral density, and echocardiography data were obtained.

Results: There was no significant difference in age, sex, duration of symptoms and thyroid function between the two groups. No serious complications occurred in any of the patients. The cost of treatment was lower in group 1 than in group 2. At the end of 10 years, goiter rate was greater and antithyroperoxidase antibody concentration was higher in group 1 than in group 2. Serum cholesterol and low density lipoprotein-cholesterol concentrations were increased in group 2 as compared with group 1; relative risks were 1.8 (1.12–2.95, P < 0.02) and 1.6 (1.09–2.34, P < 0.02) respectively. Bone mineral density and echocardiographic measurements were not different between the two groups.

Conclusion: Long-term continuous treatment of hyperthyroidism with MMI is safe. The complications and the expense of the treatment do not exceed those of radioactive iodine therapy.
Defect of a subpopulation of natural killer immune cells in Graves’ disease and Hashimoto’s thyroiditis: normalizing effect of dehydroepiandrosterone sulfate
Authors: Solerte Sebastiano Bruno , Precerutti Sara , Gazzaruso Carmine , Locatelli Eleonora , Zamboni Mauro , Schifino Nicola , Bonacasa Roberto , Rondanelli Mariangela , Taccani Davide , Ferrari Ettore , Fioravanti Marisa
Published: May 2005

Background: The study of the natural killer (NK) immune compartment could provide important findings to help in the understanding of some of the pathogenetic mechanisms related to autoimmune thyroid diseases (Graves’ disease (GD) and Hashimoto’s thyroiditis (HT)). Within this context, it was suggested that alterations in NK cell cytotoxicity (NKCC) and NK production of cytokines might occur in subjects with GD and HT, whereas the normalization of NK functions could potentially contribute to the prevention of the onset or the progression of both diseases.

Objective: Due to the hypothesis of alterations in NK in autoimmune thyroid diseases, we were interested to evaluate NKCC in GD and HT patients and to modulate NK function and secretory activity with cytokines and dehydroepiandrosterone sulfate (DHEAS) in an attempt to normalize NK cell defect.

Design: We studied 13 patients with recent onset Graves’ disease, 11 patients with Hashimoto’s thyroiditis at first diagnosis and 15 age-matched healthy subjects.

Methods: NK cells were concentrated at a density of 7.75 × 106cells/ml by negative immunomagnetic cell separation and validated by FACScan as CD16 + /CD56 + cells. NK cells were incubated with interleukin-2 (IL-2) and interferon-β (IFN-β) and co-incubated with DHEAS at different molar concentrations for measuring NKCC and the secretory pattern of tumor necrosis factor-α (TNF-α) from NK cells.

Results: Lower spontaneous, IL-2- and IFN-β-modulated NKCC was demonstrated in GD and HT patients compared with healthy subjects (P< 0.001). A decrease in spontaneous and IL-2-modulated TNF-α release from NK cells was also found in both groups of patients (P< 0.001). The co-incubation of NK cells with IL-2/IFN-β + DHEAS at different molar concentrations (from 10−8to 10−5M/ml/NK cells) promptly normalized NKCC and TNF-α secretion in GD and HT patients.

Conclusions: A functional defect of a subpopulation of NK immune cells, involving both NKCC and the secretory activity, was demonstrated in newly-diagnosed GD and HT patients. This defect can be reversed by a dose-dependent treatment with DHEAS. The impairment of NK cell activity in autoimmune thyroid diseases could potentially determine a critical expansion of T/B-cell immune compartments leading to the generation of autoantibodies and to the pathogenesis of thyroid autoimmunity.
Health status in patients with sub-clinical hypothyroidism
Authors: Razvi Salman , Ingoe Lorna E , McMillan Carolyn V , Weaver Jolanta U
Published: May 2005

Objective: Sub-clinical hypothyroidism (SCH) is a common disorder. People with this condition may have symptoms which could affect their perception of health. Therefore, the perceived health status of people with SCH was assessed and compared with population-matched norms.

Design: A prospective cross-sectional survey.

Methods: Seventy-one adults with SCH, age range 18–64 years were studied. Perceived health status was measured by the Short Form-36 (SF-36) version 2 questionaire, which has been validated in a UK population setting. The SF-36 has eight scales measuring physical functioning, role physical, bodily pain, general health, vitality, social functioning, role emotional and mental health. Their SF-36 scores were compared with UK normative data after matching for age and sex and are reported as z-scores.

Results: Scores of all eight SF-36 scales were significantly lower in people with SCH compared with the normative population. A negative score (compared with zero of the normative population) indicates worse health status. The most significantly impaired aspects of health status were vitality and role limitations due to physical problems (role physical scale) with z-scores (95% confidence intervals) of −1.01 (−0.74 to −1.29) and −0.73 (−0.43 to −1.04) respectively. Thyroid autoimmunity did not influence the results.

Conclusion: Perceived health status is significantly impaired in people with SCH when compared with UK normative population scores. This needs to be taken into consideration by clinicians when managing patients with this disease.
Systemic administration of pituitary adenylate cyclase-activating polypeptide maintains beta-cell mass and retards onset of hyperglycaemia in beta-cell-specific calmodulin-overexpressing transgenic mice
Authors: Tsunekawa Shin , Miura Yoshitaka , Yamamoto Naoki , Itoh Yuji , Ariyoshi Yoh , Senda Takao , Oiso Yutaka , Niki Ichiro
Published: May 2005

Objective: Pituitary adenylate cyclase-activating polypeptide (PACAP) has been shown to play an important role in the regulation of islet function. We investigated its effects in beta-cell-specific calmodulin-overexpressing diabetic (CaMTg) mice, in which we consider that apoptosis of beta cells is the primary defect leading to basal hyperglycaemia.

Methods: CaMTg mice were treated with continuous s.c. infusions of PACAP from 2 to 4 weeks after birth, and were evaluated against littermate non-transgenic (nTg) and saline-treated CaMTg mice as to plasma glucose levels, insulin content, islet function and morphological features.

Results: Remarkable and progressive hyperglycaemia was observed in CaMTg mice, and PACAP treatment blunted this elevation. Insulin secretion from isolated islets demonstrated an impaired response to glucose in CaMTg mice, and PACAP treatment did not cause any improvement. The total pancreatic insulin content in CaMTg mice decreased significantly to 19.1% of that in nTg mice. PACAP treatment of CaMTg mice increased the content to 158% of the value in saline-treated CaMTg mice. The insulin content in isolated islets from CaMTg mice also decreased to 15.9% of that in nTg mice, while PACAP treatment caused a doubling of the value. Immunohistochemical investigation revealed that the insulin-positive islet area was markedly smaller in CaMTg mice and that PACAP treatment significantly expanded the insulin-positive islet area.

Conclusions: These findings indicate that PACAP treatment retards the onset of hyperglycaemia in CaMTg mice by maintaining beta-cell mass and PACAP treatment may potentially be a therapeutic measure for preventing beta-cell exhaustion during hyperglycaemia.
Metformin administration improves endothelial function in women with polycystic ovary syndrome
Authors: Diamanti-Kandarakis E , Alexandraki K , Protogerou A , Piperi C , Papamichael C , Aessopos A , Lekakis J , Mavrikakis M
Published: May 2005

Objective: The aim of this study was to investigate the endothelial status in young women with polycystic ovary syndrome (PCOS), using a simple and easily reproducible hemodynamic method combined with a biological marker and to evaluate the effect of metformin treatment on these parameters.

Design: Descriptive clinical trial.

Methods: Forty young women, 20 with PCOS and 20 normal women of similar age and body mass index were studied. Metformin (1700 mg daily) was administered for 6 months to the PCOS group. The endothelium status and the metabolic and hormonal profile were studied in both groups, as well as after metformin, by flow-mediated dilatation (FMD) on the brachial artery and by measurements of plasma endothelin-1 (ET-1) levels.

Results: FMD was impaired in the PCOS group when compared with controls (3.24±0.71% vs 8.81±1.07% respectively,P< 0.0001), but this difference normalized after metformin treatment (PCOSpost-metforminvs controls: 8.17±1.26 vs 8.81±1.07%,P= 0.70) since the values significantly improved after metformin treatment (PCOSpre-metforminvs PCOSpost-metformin: 3.24±0.71 vs 8.17±1.26%,P= 0.003). ET-1 levels were significantly higher in the PCOS women compared with the control group (7.23±0.50 vs 4.99±0.69 fmol/l,P= 0.01), they improved significantly after metformin treatment (PCOSpre-metforminvs PCOSpost-metformin: 7.23±0.50 vs 3.57±0.60 fmol/l,P< 0.0001) and their difference compared with the control group was reversed (PCOSpost-metforminvs controls: 3.57±0.60 vs 4.99±0.69 fmol/l,P= 0.13). Metformin administration improved hyperandrogenemia. However, in this study, mathematical methods used to assess insulin resistance failed to show any detected alteration after treatment with metformin.

Conclusions: PCOS women were found to exhibit endothelial dysfunction compared with controls, which was reversed 6 months after metformin administration.
Fishing and Natural Mortality of Adult Largemouth Bass in a Tropical Reservoir
Authors: Waters D. Scott , Noble Richard L. , Hightower Joseph E.
Published: May 2005

Largemouth bass Micropterus salmoides, an intensively studied sport fish of temperate North America, has been introduced into tropical regions, but with little evaluation of fishing or natural mortality rates. Previous research in Puerto Rico suggested that annual mortality rates are high, but whether the extended spawning season (nearly 6 months annually), high angler harvest, or a combination was the cause of the elevated mortality was unresolved. Using ultrasonic telemetry, we tracked 44 adult largemouth bass over an 18‐month period to quantify patterns of natural, fishing, and total mortality. Using a general capture–recapture model, we estimated the annual instantaneous rates of fishing (0.584, SE = 0.164) and natural mortality (0.310, SE = 0.122). Natural mortality varied seasonally and generally increased during periods of spawning activity; however, fishing mortality was fairly consistent throughout the study. Given the relatively high fishing mortality rate, restrictions on harvest could probably be used to alter the age and size distribution of this population.
A Genetic Assessment of Ohio River Walleyes
Authors: White Matthew M. , Kassler Todd W. , Philipp David P. , Schell Scott A.
Published: May 2005

Walleyes Sander vitreus from nine Ohio River pools were examined by means of two independent restriction fragment length polymorphism (RFLP) analyses of mitochondrial DNA, allozyme analysis of nuclear DNA, and microsatellite analysis of nuclear DNA. The primary objective of this project was to document genetic variation and population structuring in walleyes among the nine pools analyzed. Both RFLP analyses revealed the presence of a distinct haplotype that was most abundant in walleyes collected from two tailwaters in the upper Ohio River (river miles 54 and 84), suggesting a remnant native population. The allozyme analysis also revealed differentiation between walleyes with the two haplotypes, even though there were no fixed differences. The microsatellite analysis indicated highly significant structuring among the nine pools analyzed. A lack of microsatellite differentiation between the two haplotypes detected in the same pool suggests that the fish are freely interbreeding. If the unique haplotype is from a remnant native stock of walleyes in the Ohio River, it could serve as a genetic marker to establish a broodstock for stocking walleyes in the upper Ohio River drainage. The second objective of the project was to evaluate the accuracy of field identifications of walleyes, saugers S. canadensis, and their hybrid, saugeyes, and to determine the direction of hybridization. Twenty‐seven percent of the fish identified as walleyes were misidentified (10% were FX hybrids and 17% F1 hybrids). All F1 hybrids were from walleye females and sauger males.
Population Viability Analysis of Atlantic Salmon in Maine, USA
Authors: Legault Christopher M.
Published: May 2005

Populations of Atlantic salmon Salmo salar in eight rivers of Maine, USA, are listed as endangered under the U.S. Endangered Species Act. This listing has required the creation of measurable and objective delisting criteria. One component of these criteria is the determination of the abundance levels associated with recovered populations. Population viability analysis (PVA) was chosen as the method to estimate these recovery levels because it (1) formalizes the combination of information available on the species. (2) quantifies the uncertainty in the population parameters and evaluates the impact of this uncertainty on the probability of extinction, and (3) allows examination of potential management strategies. A PVA specific to the Atlantic salmon populations in Maine was conducted based on the life history characteristics of the endangered populations (i.e., multiple ages of return from sea, kelting, river‐specific habitat limitations, and use of stocking as a recovery tool). The model was verified by using data from a single river and by comparing the results with those of a simpler PVA model. Projections for the eight rivers were conducted for a range of future conditions and management strategies to produce extinction probabilities that managers could use to set recovery criteria. Results beyond extinction risk, such as population age or stage structure, are also shown to be important for management consideration.
Effects of Surgically Implanted Transmitter Weights on Growth and Swimming Stamina of Small Adult Westslope Cutthroat Trout
Authors: Zale Alexander V. , Brooke Carrie , Fraser William C.
Published: May 2005

The generally accepted 2% ratio of transmitter weight to body weight constrains or precludes telemetry studies examining the timing and location of spawning of small adult westslope cutthroat trout Oncorhynchus clarkii lewisi in headwater streams. We empirically determined effects of surgically implanted dummy transmitters ranging in weight from 1 to 5 g on the swimming stamina and growth of small (81.3–206.9 g) adult westslope cutthroat trout in the laboratory to establish acceptable transmitter weights for field studies on this species. Mean growth rates and swimming stamina were not significantly different among treatments, including controls. No precipitous decline or threshold beyond which performance deteriorated markedly was observed. Data collected using telemetered westslope cutthroat trout implanted with transmitters less than about 4% of body weight should therefore approximate information about untelemetered individuals without significant bias. However, we also detected subtle effects on growth within this transmitter weight range related to individual transmitter–body weight ratios (0.5–5.3% initially), and there were indications that swimming stamina was affected similarly. Each 1% increase in transmitter– body weight ratio elicited an 11.6% decrease in growth and a possible 5.6% decrease in swimming stamina at 6 weeks postimplantation. Therefore, transmitter selection should weigh the costs of increased transmitter weight on fish performance against the benefits of longer transmission durations. In the case in which transmitter weights approaching 4% of body weight are necessary to complete a study and slight decreases in performance are not expected to affect findings materially, such weights may be acceptable. In other cases, researchers should choose the lightest possible transmitters that allow study goals to be achieved and not automatically select transmitters weighing 4% of body weight.
Heparan Sulfate of Perlecan Is Involved in Glomerular Filtration
Authors: Morita Hiroyuki , Yoshimura Ashio , Inui Kiyoko , Ideura Terukuni , Watanabe Hideto , Wang Ling , Soininen Raija , Tryggvason Karl
Published: May 2005

Perlecan is a heparan sulfate proteoglycan and a major component of the glomerular basement membrane. To understand the role of heparan sulfate chains of perlecan in glomerular filtration, detailed analyses were performed of the kidneys of

Hspg2

Δ3/Δ3 mice, whose perlecan lacks heparan sulfate attachment sites in N-terminal domain I. Macroscopic, histologic, and electron microscopic observations, as well as immunohistochemical and immunoelectron microscopic analyses using specific antibodies against perlecan and agrin core proteins, revealed no significant abnormalities in these mice under physiologic conditions. Polyethyleneimine staining demonstrated no significant changes in charge density in the glomerular basement membrane. Transcripts of other heparan sulfate proteoglycans, agrin, and collagen type XVIII, as well as perlecan, were expressed at similar levels to those in the wild-type littermates. Approximately 40% of the perlecan synthesized by

Hspg2

Δ3/Δ3 fibroblasts was substituted with heparin sulfate and 60% was substituted with chondroitin sulfate. All of the perlecan synthesized by wild-type fibroblasts contained heparin sulfate, indicating an altered substitution of glycosaminoglycans on

Hspg2

Δ3/Δ3 perlecan. Immunostaining indicated that the level of chondroitin sulfate was actually increased in the

Hspg2

Δ3/Δ3 glomerular basement membrane. When administered intraperitoneally with BSA,

Hspg2

Δ3/Δ3 mice exhibited remarkable proteinuria. These findings suggest that heparan sulfate chains of perlecan play an important role in glomerular filtration, especially of a large amount of protein.
Integration of Embryonic Stem Cells in Metanephric Kidney Organ Culture
Authors: Steenhard Brooke M. , Isom Kathryn S. , Cazcarro Patricia , Dunmore Judy H. , Godwin Alan R. , St. John Patricia L. , Abrahamson Dale R.
Published: May 2005

Many stages of nephrogenesis can be studied using cultured embryonic kidneys, but there is no efficient technique available to readily knockdown or overexpress transgenes for rapid evaluation of resulting phenotypes. Embryonic stem (ES) cells have unlimited developmental potential and can be manipulated at the molecular genetic level by a variety of methods. The aim of this study was to determine if ES cells could respond to developmental signals within the mouse embryonic day 12 to embryonic day 13 (E12 to E13) kidney microenvironment and incorporate into kidney structures. ROSA26 ES cells were shown to express β-galactosidase ubiquitously when cultured in the presence of leukemia inhibitory factor to suppress differentiation. When these cells were microinjected into E12 to E13 metanephroi and then placed in transwell organ culture, ES cell-derived, β-galactosidase-positive cells were identified in epithelial structures resembling tubules. On rare occasions, individual ES cells were observed in structures resembling glomerular tufts. Electron microscopy showed that the ES cell-derived tubules were surrounded by basement membrane and had apical microvilli and junctional complexes. Marker analysis revealed that a subset of these epithelial tubules bound

Lotus tetragonolobus

and expressed α1 Na+/K+ ATPase. ES cells were infected before injection with a cytomegalovirus promoter-green fluorescence protein (GFP) adenovirus and GFP expression was found as early as 18 h, persisting for up to 48 h in cultured kidneys. This ES cell technology may achieve the objective of obtaining a versatile cell culture system in which molecular interventions can be used

in vitro

and consequences of these perturbations on the normal kidney development program

in vivo

can be studied.
Importance of Baseline Distribution of Proteinuria in Renal Outcomes Trials: Lessons from the Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) Study
Authors: Zhang Zhongxin , Shahinfar Shahnaz , Keane William F. , Ramjit Denise , Dickson Tania Z. , Gleim Gilbert W. , Mogensen Carl Erik , de Zeeuw Dick , Brenner Barry M. , Snapinn Steven M.
Published: May 2005

A key issue in the analysis of outcome trials is the adjustment for baseline covariates that influence the primary outcome. Imbalance of an important covariate between treatment groups at baseline is of considerable concern if one treatment group is favored over another with respect to the hypothesis testing outcome. With the use of the Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) study database as an example, the influence of baseline proteinuria on the primary composite endpoint, ESRD, and ESRD or death after adjusting for baseline proteinuria as a continuous covariate was examined. Increasing baseline proteinuria was associated with increased risk for renal events, confirming that proteinuria is an important covariate for renal outcomes. When the randomization was stratified according proteinuria <2000 mg/g or ≥2000 mg/g, within the higher proteinuria stratum (≥2000 mg/g), patients in the losartan group had a higher baseline mean proteinuria value. When the imbalance was adjusted, an increase in the magnitude and the significance of the risk reduction with losartan for each outcome was observed. No apparent interaction between treatment effect and baseline proteinuria was found, and there was no heterogeneity in the treatment response in patients with different baseline proteinuria levels. After proteinuria was adjusted as a continuous variable, greater treatment effects were observed in the RENAAL study. This effect was due solely to the imbalance in baseline proteinuria. Considering the importance of proteinuria as a risk factor, adjustment for baseline proteinuria as a continuous covariate should be prespecified in the design and analysis of clinical trials involving renal outcomes, even when patients are stratified on the basis of level of proteinuria.
Detection and Localization of Proteinuria by Dynamic Contrast-Enhanced Magnetic Resonance Imaging Using MS325
Authors: Zhang Yantian , Choyke Peter L. , Lu Huiyan , Takahashi Hideko , Mannon Roslyn B. , Zhang Xiaojie , Marcos Hani , Li King C.P. , Kopp Jeffrey B.
Published: May 2005

After renal transplantation, persistent glomerular disease affecting the native kidneys typically causes albuminuria, at least for a period of time, making it difficult to determine in a noninvasive fashion whether proteinuria originates in the native kidneys or the renal allograft. To address this problem, dynamic contrast-enhanced magnetic resonance imaging (MRI) using gadolinium (Gd)-based albumin-bound blood pool contrast agent (MS325) to localize proteinuria was investigated. Glomerular proteinuria was induced in Sprague-Dawley rats by intravenous injection of puromycin aminonucleoside (PAN), whereas control rats received physiologic saline vehicle. Both groups of animals underwent a 40-min dynamic contrast-enhanced MRI using radio frequency spoiled gradient echo imaging sequence after injection of Gd-labeled MS325. Contrast uptake and clearance curves for cortex and medulla were determined from acquired MR images. Compared with controls, proteinuric rats exhibited significantly lower elimination rate constants. The use of gadopentetate dimeglumine (Gd-DTPA) as a contrast agent showed smaller and less specific differences between proteinuric and control groups. In rats with one proteinuric kidney (PAN-treated) and one normal kidney (transplanted from a normal rat), MRI using MS325 was able to differentiate between the two kidneys. The results suggest that MRI with an albumin-bound blood pool contrast agent may be a useful noninvasive way to localize proteinuria. If this technique can be successfully applied in human patients, it may allow for the localization of proteinuria after kidney transplant and thereby provide a noninvasive way to detect disease affecting the renal allograft.