Disease modifying effect of statins in dextran sulfate sodium model of mouse colitis

Objective: We evaluated the disease modifying effect of simvastatin and atorvastatin in Dextran Sulfate Sodium (DSS) model of colitis. Materials and methods: Thirty, 8-week old female Swiss-Webster mice were separated into 5 groups (n = 6/group). Colitis was induced by feeding 4 % DSS solution for 7 days. Following discontinuation of DSS, over the next 7 days, the groups orally received simvastatin (20 mg/kg/day), atorvastatin (60 mg/kg/day), vehicle only (0.75 % methylcellulose), subcutaneous 30 μg injections of anti-TNFα monoclonal antibody or intraperitoneal anti-mouse apolipoprotein A-I antibody respectively. Disease activity Index (DAI) was determined daily by a blinded investigator. Results: The mean reduction in DAI scores from day 7 to day 14 for anti-TNFα group, simvastatin and atorvastatin group were 74 %, 76 % and 64 %, respectively as compared to 41 % reduction in vehicle and anti-apolipoprotein A-I antibodytreated groups. Conclusions: This finding suggests that statins may have the ability to modify the disease activity in the DSS model of colitis and the disease modifying effect is comparable to anti-mouse TNFα treatment in this model. Received 23 October 2006; returned for revision 27 February 2007; accepted by I. Ahnfelt-R?nne 16 August 2007     

Chromosome pairing in allotetraploid hybrids of <Emphasis Type="Italic">Festuca pratensis</Emphasis> × <Emphasis Type="Italic">Lolium perenne</Emphasis> revealed by genomic <Emphasis Type="Italic">in situ</Emphasis> hybridization (GISH)

Genomic in situ hybridization (GISH) was used to make a detailed study of chromosome pairing at metaphase I (MI) of meiosis in six F(1) hybrid plants of the allotetraploid Festuca pratensis x Lolium perenne (2n = 4x = 28; genomic constitution FpFpLpLp). The mean chromosome configurations for all hybrids analysed were 1.13 univalents + 11.51 bivalents + 0.32 trivalents + 0.72 quadrivalents, and the mean chiasma frequency was 21.96 per cell. GISH showed that pairing was predominantly intragenomic, with mean numbers of L. perenne (Lp/Lp) and F. pratensis (Fp/Fp) bivalents being virtually equal at 5.41 and 5.48 per cell, respectively. Intergenomic pairing between Lolium and Festuca chromosomes was observed in 33.3% of Lp/Fp bivalents (0.62 per cell), in 79.7% of trivalents - Lp/Lp/Fp and Lp/Fp/Fp (0.25 per cell), and in 98.4% of quadrivalents - Lp/Lp/Fp/Fp and Lp/Lp/Lp/Fp (0.71 per cell). About 4.0% of the total chromosome complement analysed remained as univalents, an average 0.68 Lp and 0.45 Fp univalents per cell. It is evident that in these hybrids there is opportunity for recombination to take place between the two component genomes, albeit at a low level, and this is discussed in the context of compromising the stability of Festulolium hybrid cultivars and accounting for the drift in the balance of the genomes over generations. We speculate that genotypic differences between hybrids could permit selection for pairing control, and that preferences for homologous versus homoeologous centromeres in their spindle attachments and movement to the poles at anaphase I could form the basis of a mechanism underlying genome drift.     

Topical acetone treatment induces neurogenic oedema on the sensitized mouse ear: an <Emphasis Type="Italic">in vivo</Emphasis> study using transient receptor potential vanilloid 1 (TRPV1) receptor knockout mice

Objective: The participation of sensory neurons and transient receptor potential vanilloid 1 (TRPV1) receptors in phorbol 12-myristate 13-acetate (PMA)-induced nerve-sensitizing effect was examined. Materials and methods: PMA dissolved in acetone and acetone were applied to the ears of TRPV1 receptor knockout and wild-type mice. Different groups of animals received ibuprofen, anti-interleukin-1 beta (IL-1β) antibody, resiniferatoxin (RTX) or capsaicin pretreatment. Ear thickness, myeloperoxidase activity and IL-1β content of the ears were determined. Histological evaluation was performed. Results: PMA exerted potentiating action on contralateral acetone-induced ear oedema, which was inhibited by ibuprofen, topical capsaicin desensitization of the acetone-treated ear as well as by systemic RTX pretreatment. Neither the lack of TRPV1 receptors nor anti-IL-1β antibody prevented sensitizing effect. Conclusions: The TRPV1 receptor-independent potentiating action of PMA on contralateral acetone-induced ear oedema is mediated via capsaicin-sensitive afferents and prostanoids are involved. IL-1β is not essential in this process. Received 3 April 2007; returned for revision 31 May 2007; accepted by G. Geislinger 8 June 2007     

Clinical dermatitis in a southern brown bandicoot (<Emphasis Type="Italic">Isoodon obesulus</Emphasis>) associated with the mite <Emphasis Type="Italic">Sarcoptes scabiei</Emphasis>

A wild-caught adult male southern brown bandicoot (Isoodon obesulus) was presented for investigation of a pruritic skin condition that consisted of crusting, deep fissures, lichenification, alopecia, scale, and erythema, and affected the caudal dorsum, caudal, and medial thighs, distal hind limbs, tail, forepaws, and parts of the thorax. Examination of superficial and deep skin scrapings revealed large numbers of mites, which were identified as Sarcoptes scabiei. To the authors’ knowledge, infestation with this mite has not previously been reported in bandicoots.     

Inflammatory fibroid polyp of the small bowel with a mutation in exon 12 of <Emphasis Type="Italic">PDGFRα</Emphasis>

Inflammatory fibroid polyp (IFP) is a benign reactive uncommon submucosal lesion of the gastrointestinal tract, the small intestine being the most common site of origin. Histologically, IFPs are characterized by spindle cells, a heavy inflammatory infiltrate including eosinophils and onion-sheet-like formation of lesional cells around blood vessels. We present a case report of an IFP harboring an activation mutation in the PDGFRα gene. The lesion was positive for CD34, PDGFRα, and p-PDGFRα immunostaining but was negative for c-KIT and desmin. After a sequencing analysis of KIT and PDGFRα, a mutation consisting of an in-frame deletion of codons 567-571 and a missense mutation in codon 566 (S566R) of PDGFRα was observed. This mutation could activate key cellular pathways with involvement in the pathogenesis of this entity. We concluded that more studies are necessary in order to clarify if this finding is a biologically distinct behavior or, on the contrary, represents a specific feature of the IFP.     

<Emphasis Type="Italic">In Vivo</Emphasis> Effect of Selective Macrophage Suppression on the Development of Intrahepatic Cholestasis in Mice

We studied the role of selective suppression of liver Kupffer cells (gadolinium chloride, 14 mg/kg intravenously) in the development of intrahepatic cholestasis in CBA/C57Bl/6 mice after intraperitoneal injection of α-naphthylisothiocyanate in a single dose of 200 mg/kg. Pretreatment with gadolinium chloride increased the severity of cholestasis and signs of liver damage. Gadolinium accumulation in the liver peaked after 24 h and was accompanied by a decrease in activities of cathepsin D and cathepsin B and concentration of matrix metalloprotease-2. Our results confirm the hypothesis that normal function of Kupffer cells and extracellular matrix plays an important role in cholestasis. Administration of gadolinium chloride serves as a convenient model to study the side effects, toxicity, and safety of lanthanides as nanoparticles. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 146, No. 10, pp. 376–380, October, 2008     

Chromosome homology between mouse and three Muridae species, <Emphasis Type="Italic">Millardia meltada</Emphasis>, <Emphasis Type="Italic">Acomys dimidiatus</Emphasis> and <Emphasis Type="Italic">Micromys minutus</Emphasis>, and conserved chromosome segments in murid karyotypes

Comparative chromosome painting with mouse (Mus musculus, MMU) chromosome-specific DNA probes was performed for three Muridae species, the Indian soft-furred field rat (Millardia meltada), the spiny mouse (Acomys dimidiatus) and the harvest mouse (Micromys minutus). All probes except for the Y probe were successfully hybridized to the chromosomes of all species, and homologous chromosome segments between mouse and the three species were identified at the molecular level. Comparison of our data with the published data of six other genera (Mus, Rattus, Apodemus, Otomys, Rhabdomys and Cricetulus) of the Muridae suggested that the associations MMU1b/17a, 2b/13a, 5b/11a, 7/19, 10b/17b, 10c/17c, 11b/16a, 12/17d and 13b/15, and the single painted chromosomes and chromosome segments MMU3, 4, 5a, 8a, 8b, 16b, 18 and X were probably contained by the ancestral karyotype of the Muridae, and have been strongly conserved throughout murid evolution.     

Histopathologic and electron microscopic characterization of cutaneous leishmaniasis in <Emphasis Type="Italic">Tatera indica</Emphasis> and <Emphasis Type="Italic">Gerbillus</Emphasis> spp. infected with <Emphasis Type="Italic">Leishmania major</Emphasis>

Cutaneous leishmaniasis (CL) is a zoonotic disease transmitted between rodents and canines, mainly by Phlebotomus sand flies and man. In southern Iran, the incidence of this protozoan disease has doubled over the last decade. The present study deals with histopathological features of CL in Tatera indica and Gerbillus spp. that participate in the epidemiology of CL in southern Iran. Thirty-two trapped rodents were evaluated for any Leishmania infection using enzyme electrophoresis and polymerase chain reaction and were concomitantly studied for any histopathological changes. Histopathological studies showed that bone marrow was the tissue of choice for light and electron microscopic study of Leishmania, demonstrating the macrophages infected with the amastigote form of the parasite. This is the first report of the histopathological detection of L. major in naturally infected T. indica and Gerbillus spp in the Larestan region.     

C-<Emphasis Type="Italic">erb</Emphasis>B-2 oncogene and <Emphasis Type="Italic">P21</Emphasis><Superscript><Emphasis Type="Italic">WAF/CIP1</Emphasis></Superscript> tumor suppressor gene expression as prognostic factors in canine mammary adenocarcinomas

To evaluate the use of c-erbB-2 oncogene and p21 ^WAF1/CIP1 suppressor gene products as the prognosis markers for canine mammary tumors, expression of these gene products were examined immunohistochemically using tumor tissues and clinical data from 96 dogs with malignant mammary tumors. Semiquantitative data was compared with histopathological grades, proliferating cell nuclear antigen (PCNA)-positive index, and clinicopathological matters. The expression c-erbB-2 protein was found in the cellular membrane and cytoplasm of neoplastic epithelial cells, and the positive index had no significant relation to the histopathological features and PCNA-positive index, except for the individual age of affected dogs (P < 0.05). The product of p21 ^WAF1/CIP1 was mostly found in cytoplasm and occasionally in the nucleus of neoplastic cells. The quantitative data had significant association to the malignancy grade and size of tumors (P < 0.05). However, that had no significant relationship to the PCNA-positive index. The present study concluded that both gene products could not apply as the direct markers to evaluate the prognosis of canine mammary tumors. The detection of c-erbB-2 product may be partly beneficial to the differential diagnosis of epithelial type of mammary cancer. The use of p21 ^WAF1/CIP1 product in prognosis of canine mammary cancer needs further investigation.     

Haematology and Serum Biochemistry of the Tammar Wallaby, <Emphasis Type="BoldItalic">Macropus eugenii</Emphasis>

Haematological and serum biochemical data have been collected from a total of 104 tammar wallabies maintained in a captive population at Macquarie University, NSW, Australia. These data have been analysed with respect to the effects of age, sex and seasonality. Age-related effects were apparent for both haematological parameters and serum biochemistry. Erythrocyte parameters, including haemoglobin, total erythrocyte count, haematocrit, macrocyte volume index and mean corpuscular haemaglobin, but not the leucocyte parameters, total leucocyte count, neutrophils, lymphocytes, monocytes, eosinophils, platelets and maximum packed volume, differed between the sexes. Seasonal effects were apparent in both leucocyte and erythrocyte parameters. Serum parameters of total protein and bilirubin were higher in autumn whereas alkaline phosphatase values were higher in summer. These data will serve as an effective management tool for assessing and monitoring the health status of individuals, particularly those in captivity. Correspondence and offprint requests to: E. M. Deane, Division of Environmental and Life Sciences, Macquarie University, North Ryde, NSW 2109, Australia. Tel: 61 2 9850 8418; Fax: 61 2 9850 9671; e-mail: edeane@els.mq.edu.au     

Comparison of <Emphasis Type="Italic">SYBR Green I</Emphasis> and <Emphasis Type="Italic">TaqMan</Emphasis> real-time PCR formats for the analysis of <Emphasis Type="Italic">her2</Emphasis> gene dose in human breast tumors

We compared two technologies of real-time PCR (with the use of fluorescent SYBR Green I dye and specific TaqMan probe) for quantification of the dose of her2 gene in breast tumors. The maximum increase in the gene dose in TaqMan and SYBR Green I analyses was 10-and 5-fold, respectively. In was found that TaqMan and SYBR Green I technologies allow detection of the matrix in amounts corresponding to 1–100 and 2.5–40.0 ng genomic DNA, respectively. Tenfold increase in the gene dose leads to incorrect evaluation of multiplication ratio in the SYBR Green I analysis. These results suggest that TaqMan technology is more preferable for correct evaluation of her2 gene dose. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 2, pp. 201–205, February, 2008     

Comparative Nuclear Magnetic Resonance Studies of Diffusional Water Permeability of Red Blood Cells from Different Species. XII. Dog (<Emphasis Type="BoldItalic">Canis familiaris</Emphasis>) and Cat (<Emphasis Type="BoldItalic">Felis domestica</Emphasis>)

The diffusional water permeability (P _d ) of dog and cat red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition with p-chloromercuribenzene sulphonate (PCMBS). The values of P _d were in the case of cat RBC ∼3.0 × 10⁻³ cm/s at 15 °C, 3.5 × 10⁻³ cm/s at 20 °C, 4.2 × 10⁻³ cm/s at 25 °C, 4.4 × 10⁻³ cm/s at 30 °C and 5.9 × 10⁻³ cm/s at 38 °C. In case of dog RBC the values of P _d were higher ∼3.8 × 10⁻³ cm/s at 15 °C, 4.6 × 10⁻³ cm/s at 20 °C, 5.0 × 10⁻³ cm/s at 25 °C, 5.9 × 10⁻³ cm/s at 30 °C and 7.9 × 10⁻³ cm/s at 37 °C. Systematic studies of the effect of PCMBS on water diffusion indicated that in the case of dog RBCs the maximal inhibition was reached in 15–30 min with 1 mm PCMBS, whereas in the case of cat RBCs in 60 min with 1 mm PCMBS or in 30 min with 2 mm PCMBS. The values of maximal inhibition in the case of cat RBC were in the range of 55–60% at 15 °C, 60–68% at 20 °C and 25 °C, 50–60% at 30 °C and 50–55% at 37 °C. In the case of dog RBC the corresponding values were higher, 75–80% at 15 °C, 70–80% at 20 °C and 25 °C, 65–70% at 30 °C and 55–60% at 37 °C. The basal permeability to water was estimated to be ∼1 × 10⁻³ cm/s −2 × 10⁻³ cm/s in the range of temperatures of 25–37 °C. The activation energy of water diffusion E _a,d was ∼19 kJ/mol for the dog RBC and ∼23 kJ/mol for the cat RBC. After incubation with PCMBS the values of E _a,d increased, reaching 40 kJ/mol in conditions of maximal inhibition of water exchange. The membrane polypeptide electrophoretic pattern of dog and cat RBCs has been compared with its human counterpart. Dog and cat RBCs contained higher amounts of spectrin (band 1 and 2) and lower amounts of bands 4.4, 4.2, band 5 and band 7 compared to human RBCs. Band 4.9 was decreased only in the cat RBCs, whereas band 6 was decreased only in the dog RBCs. Correspondence and offprint requests to: Gheorghe Benga, Department of Cell and Molecular Biology, ‘Iuliu Haţieganu’ University of Medicine and Pharmacy, 6 Pasteur St, 3400 Cluj-Napoca, Romania. Tel:/Fax: 40–64–194373; e-mail: GBenga@personal.ro; gbenga@umfcluj.ro     

Catalog of 86 single-nucleotide polymorphisms (SNPs) in three uridine diphosphate glycosyltransferase genes: <Emphasis Type="Italic">UGT2A1</Emphasis>, <Emphasis Type="Italic">UGT2B15</Emphasis>, and <Emphasis Type="Italic">UGT8</Emphasis>

We report here three high-density maps of variations found among 48 Japanese individuals in three uridine diphosphate glycosyltransferase (UGT) genes, UGT2A1, UGT2B15, and UGT8. A total of 86 single-nucleotide polymorphisms (SNPs) were identified through systematic screening of genomic regions containing these genes: 8 in 5′ flanking regions, 7 in coding regions, 67 in introns, 3 in 3′ untranslated regions, and 1 in a 3′ flanking region. We also discovered 14 variations of other types. Of the 86 SNPs, 63 (73%) were considered to be novel on the basis of comparison of our data with the Database of SNPs (dbSNP) of the National Center for Biotechnology Information. Among the seven SNPs identified in exonic sequences, five were non-synonymous changes that would result in amino-acid substitutions. The collection of SNPs derived from this study will serve as an additional resource for studies of complex genetic diseases and responsiveness to drug therapy. Received: June 12, 2002 / Accepted: June 13, 2002     

Use of cross-species <Emphasis Type="Italic">in-situ</Emphasis> hybridization (ZOO-FISH) to assess chromosome abnormalities in day-6 <Emphasis Type="Italic">in-vivo</Emphasis>- or <Emphasis Type="Italic">in-vitro</Emphasis>-produced sheep embryos

Causes of chromosomal differences such as mosaicism between embryos developed in vivo and in vitro may be resolved using animal models to compare embryos generated in vivo with those generated by different production systems. The aims of this study were: (1) to test a ZOO-FISH approach (using bovine painting probes) to detect abnormal chromosome make-up in the sheep embryo model, and (2) to examine the extent of chromosome deviation in sheep embryos derived in vivo and in vitro. Cytogenetic analysis was performed on day 6 in-vivo and in-vitro derived sheep embryos using commercially available bovine chromosome painting probes for sex chromosomes X–Y and autosomes 1–29. A total of 8631 interphase and metaphase nuclei were analyzed from 49 in-vitro-derived and 51 in-vivo-derived embryos. The extent of deviation from normal ovine chromosome make-up was higher (p < 0.05) in in-vitro-produced embryos relative to in-vivo-derived embryos (65.3% vs. 19.6% respectively) mainly due to diploid–polyploid mosaicism. Polyploid cells ranged from 3n to 8n with tetraploids most predominant among non-diploid cells. The proportions of polyploid cells per mixoploid embryo in in-vitro-produced embryos ranged from 1.4% to 30.3%, in contrast to less than 10% among the in-vivo-derived embryos. It was concluded that in-vitro-derived embryos are vulnerable to ploidy change compared to their in-vivo counterparts. The application of ZOO-FISH to domestic animal embryos is an effective approach to study the chromosome complement of species for which DNA probes are unavailable.     

A frameshift mutation of the chloroplast <Emphasis Type="Italic">mat</Emphasis>K coding region is associated with chlorophyll deficiency in the <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> virescent mutant <Emphasis Type="Italic">Wogon</Emphasis>-Sugi

Wogon-Sugi has been reported as a cytoplasmically inherited virescent mutant selected from a horticultural variety of Cryptomeria japonica. Although previous studies of plastid structure and inheritance indicated that at least some mutations are encoded by the chloroplast genome, the causative gene responsible for the primary chlorophyll deficiency in Wogon-Sugi, has not been identified. In this study, we identified this gene by genomic sequencing of chloroplast DNA and genetic analysis. Chloroplast DNA sequencing of 16 wild-type and 16 Wogon-Sugi plants showed a 19-bp insertional sequence in the matK coding region in the Wogon-Sugi. This insertion disrupted the matK reading frame. Although an indel mutation in the ycf1 and ycf2 coding region was detected in Wogon-Sugi, sequence variations similar to that of Wogon-Sugi were also detected in several wild-type lines, and they maintained the reading frame. Genetic analysis of the 19 bp insertional mutation in the matK coding region showed that it was found only in the chlorophyll-deficient sector of 125 full-sibling seedlings. Therefore, the 19-bp insertion in the matK coding region is the most likely candidate at present for a mutation underlying the Wogon-Sugi phenotype. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Meiotic recombination in <Emphasis Type="Italic">Caenorhabditis elegans</Emphasis>

The faithful segregation of homologous chromosomes during meiosis is dependent on the formation of physical connections (chiasma) that form following reciprocal exchange of DNA molecules during meiotic recombination. Here we review the current knowledge in the Caenorhabditis elegans meiotic recombination field. We discuss recent developments that have improved our understanding of the crucial steps that must precede the initiation and propagation of meiotic recombination. We summarize the pathways that impact on meiotic prophase entry and the current understanding of how chromosomes reorganize and interact to promote homologous chromosome pairing and subsequent synapsis. We pay particular attention to the mechanisms that contribute to meiotic DNA double-strand break (DSB) formation and strand exchange processes, and how the C. elegans system compares with other model organisms. Finally, we highlight current and future areas of research that are likely to further our understanding of the meiotic recombination process.     

Evaluation of hematological values in indigenous chickens infected with <Emphasis Type="Italic">Plasmodium gallinaceum</Emphasis> and <Emphasis Type="Italic">Aegyptianella pullorum</Emphasis>

Plasmodium, Haemoproteus, and leukocytozoon are the most important hematozoa in birds, which have been reported in different areas of the world. The present study was undertaken to find which blood protozoans exist in indigenous chickens in Shiraz, southern Iran and to evaluate hematological parameters in birds infected with hematozoas. Plasmodium and Aegyptianella were the two parasites found in 740 blood samples examined from indigenous chickens of which 29 (3.91%) were positive for Aegyptianella pullorum, 106 (14.32%) for Plasmodium gallinaceum, and 12 (1.62%) for A. pullorum and Plasmodium gallinaceum together. There was no significant difference between hematological parameters of non-infected and naturally infected chickens with Plasmodium gallinaceum, A. pullorum, and both (P > 0.05). Low infection of indigenous chickens with A. pullorum, Plasmodium gallinaceum, and both had no significant effects on hematological parameters (P > 0.05), which is probably due to low parasitemia rate and immunity against these two parasites.     

<Emphasis Type="Italic">Methionine synthase A2756G</Emphasis> polymorphism may predict ulcerative colitis and <Emphasis Type="Italic">methylenetetrahydrofolate reductase C677T</Emphasis> pancolitis,in Central China

Background  

The association of genetic polymorphisms related to metabolism of homocysteine with inflammatory bowel disease has been evidenced in Crohn disease and remains an open question in ulcerative colitis. We evaluated the association of the polymorphisms of MTHFR, MTR, MTRR and TCN2 genes with ulcerative colitis in Central China.