bluuebunny/crossref_metadata_2025 huggingface dataset:
Complete sequence of a duck astrovirus associated with fatal hepatitis in ducklings
Authors: Fu Yu , Pan Meng , Wang Xiaoyan , Xu Yongliang , Xie Xiaoyu , Knowles Nick J. , Yang Hanchun , Zhang Dabing
Published: April 2009

Duck astroviruses (DAstVs) are known to cause duck viral hepatitis; however, little is known regarding their molecular biology. Here, we report the complete sequence of a DAstV associated with a recent outbreak of fatal hepatitis in ducklings in China. Sequence analyses indicated that the genome of DAstV possessed a typical astrovirus organization and also exhibited two unique features. The polyadenylated genome comprised 7722 nt, which is the largest among astroviruses sequenced to date. The ORF2 of DAstV was not in the same reading frame as either ORF1a or ORF1b, which was distinct from all other astroviruses. Sequence comparisons and phylogenetic analyses revealed that DAstV was more closely related to turkey astrovirus (TAstV) type 2, TAstV-3 and TAstV/MN/01 (a possible new TAstV serotype) than to TAstV-1 or other astroviruses. These findings suggest that astroviruses may transmit across ducks and turkeys.
Herpes simplex virus type 2 tegument proteins contain subdominant T-cell epitopes detectable in BALB/c mice after DNA immunization and infection
Authors: Muller William J. , Dong Lichun , Vilalta Adrian , Byrd Benjamin , Wilhelm Kai M. , McClurkan Christopher L. , Margalith Michal , Liu Chao , Kaslow David , Sidney John , Sette Alessandro , Koelle David M.
Published: April 2009

Cytotoxic T cells are important in controlling herpes simplex virus type 2 (HSV-2) reactivation and peripheral lesion resolution. Humans latently infected with HSV-2 have cytotoxic T cells directed against epitopes present in tegument proteins. Studies in mice of immunity to HSV have commonly focused on immunodominant responses in HSV envelope glycoproteins. These antigens have not proved to be an effective prophylactic vaccine target for most of the human population. The murine immune response against HSV tegument proteins has not been explored. We analysed cellular responses in BALB/c mice directed against the tegument proteins encoded byUL46, UL47andUL49and against the envelope glycoprotein gD after DNA vaccination or HSV-2 infection. After DNA vaccination, the splenocyte T-cell response to overlapping peptides fromUL46andUL47was more than 500 gamma interferon spot-forming units per 106responder cells. Peptide truncation studies, responder cell fractionation and major histocompatibility complex binding studies identified several CD8+and CD4+epitopes. Cellular responses to tegument protein epitopes were also detected after HSV-2 infection. Tegument proteins are rational candidates for further HSV-2 vaccine research.
Treatment with respiratory syncytial virus G glycoprotein monoclonal antibody or F(ab′)2 components mediates reduced pulmonary inflammation in mice
Authors: Miao Congrong , Radu Gertrud U. , Caidi Hayat , Tripp Ralph A. , Anderson Larry J. , Haynes Lia M.
Published: April 2009

Therapeutic treatment with a non-neutralizing monoclonal antibody (mAb) (131-2G) specific to respiratory syncytial virus (RSV) G glycoprotein mediates virus clearance and decreases leukocyte trafficking and interferon gamma (IFN-γ) production in the lungs of RSV-infected mice. Its F(ab′)2 component only mediates decreased leukocyte trafficking and IFN-γ production without reducing virus replication. Thus, this mAb has two independent actions that could facilitate treatment and/or prevention of RSV infection by reducing both virus replication and virus-induced pulmonary inflammation.
Analysis of promoter activity of selected Cotesia plutellae bracovirus genes
Authors: Choi Jae Young , Kwon Soo-Jin , Roh Jong Yul , Yang Tae-Jin , Li Ming Shun , Park Beom-Seok , Kim Yonggyun , Woo Soo-Dong , Jin Byung Rae , Je Yeon Ho
Published: April 2009

In a previous study, we cloned 27 discrete genome segments ofCotesia plutellaebracovirus (CpBV) and provided the complete nucleotide sequences and annotation. Seven putative coding regions were predicted from one of the largest segments, CpBV-S30. The activity of promoters associated with six predicted ORFs from this segment were investigated using both transient and baculovirus expression assays with enhanced green fluorescent protein as a reporter gene. CpBV promoters showed activity earlier than thepolyhedrinpromoter and the activity of some of these promoters was superior to that of theAutographa californicamultiple nucleopolyhedrovirus (AcMNPV)ie-1promoter in the baculovirus expression assays. The promoter of ORF3004 showed the highest level of activity in insect cells, exhibiting 24 % of the activity obtained with the AcMNPVpolyhedrinpromoter in Sf9 cells. InSpodoptera exigualarvae, the ORF3006 promoter showed the highest activity, with about 35 % of the activity measured with thepolyhedrinpromoter. In addition, analysis of the ORF3006 promoter revealed that the region between −382 and −422 from the translation start point was critical for activity of this promoter. These results suggest that the CpBV-S30 promoters characterized here could be useful tools in a variety of biotechnological applications, such as gene expression analyses and insecticide development.
Analysis of transcripts from predicted open reading frames of Musca domestica salivary gland hypertrophy virus
Authors: Salem Tamer Z. , Garcia-Maruniak Alejandra , Lietze Verena-U. , Maruniak James E. , Boucias Drion G.
Published: April 2009

TheMusca domesticasalivary gland hypertrophy virus (MdSGHV) is a large dsDNA virus that infects and sterilizes adult houseflies. The transcriptome of this newly described virus was analysed by rapid amplification of cDNA 3′-ends (3′-RACE) and RT-PCR. Direct sequencing of 3′-RACE products revealed 78 poly(A) transcripts containing 95 of the 108 putative ORFs. An additional six ORFs not amplified by 3′-RACE were detected by RT-PCR. Only seven of the 108 putative ORFs were not amplified by either 3′-RACE or RT-PCR. A series of 5′-RACE reactions were conducted on selected ORFs that were identified by 3′-RACE to be transcribed in tandem (tandem transcripts). In the majority of cases, the downstream ORFs were detected as single transcripts as well as components of the tandem transcripts, whereas the upstream ORFs were found only in tandem transcripts. The only exception was the upstream ORF MdSGHV084, which was differentially transcribed as a single transcript at 1 and 2 days post-infection (days p.i.) and as a tandem transcript (MdSGHV084/085) at 2 days p.i. Transcriptome analysis of MdSGHV detected splicing in the 3′ untranslated region (3′-UTR) and extensive heterogeneity in the polyadenylation signals and cleavage sites. In addition, 23 overlapping antisense transcripts were found. In conclusion, sequencing the 3′-RACE products without cloning served as an alternative approach to detect both 3′-UTRs and transcript variants of this large DNA virus.
Comparative inference of illegitimate recombination between rice and sorghum duplicated genes produced by polyploidization
Authors: Wang Xiyin , Tang Haibao , Bowers John E. , Paterson Andrew H.
Published: April 2009

Whole-genome duplication produces massive duplicated blocks in plant genomes. Sharing appreciable sequence similarity, duplicated blocks may have been affected by illegitimate recombination. However, large-scale evaluation of illegitimate recombination in plant genomes has not been possible previously. Here, based on comparative and phylogenetic analysis of the sequenced genomes of rice and sorghum, we report evidence of extensive and long-lasting recombination between duplicated blocks. We estimated that at least 5.5% and 4.1% of rice and sorghum duplicated genes have been affected by nonreciprocal recombination (gene conversion) over nearly their full length after rice–sorghum divergence, while even more (8.7% and 8.1%, respectively) have been converted over portions of their length. We found that conversion occurs in higher frequency toward the terminal regions of chromosomes, and expression patterns of converted genes are more positively correlated than nonconverted ones. Though converted paralogs are more similar to one another than nonconverted ones, elevated nucleotide differences between rice–sorghum orthologs indicates that they have evolved at a faster rate, implying that recombination acts as an accelerating, rather than a conservative, element. The converted genes show no change in selection pressure. We also found no evidence that conversion contributed to guanine-cytosine (GC) content elevation.
Systemic NK4 gene therapy inhibits tumor growth and metastasis of melanoma and lung carcinoma in syngeneic mouse tumor models
Authors: Kishi Yuko , Kuba Keiji , Nakamura Takahiro , Wen Jinhua , Suzuki Yoshinori , Mizuno Shinya , Nukiwa Toshihiro , Matsumoto Kunio , Nakamura Toshikazu
Published: April 2009

Hepatocyte growth factor (HGF) promotes malignant development of cancer cells by enhancing invasion and metastasis. NK4, a competitive antagonist for HGF, is a bifunctional molecule that acts as a HGF antagonist and angiogenesis inhibitor. Although successful tumor inhibition by NK4 gene expression in tumor models has been demonstrated, the effects of systemic NK4 gene introduction are yet to be addressed. Here we show that systemic administration of a replication‐defective adenovirus expressing NK4 (Ad.NK4) inhibits tumor growth and lung metastasis of B16F10 melanoma and Lewis lung carcinoma in syngeneic mice. Single tail‐vein injection of Ad.NK4 achieved therapeutic levels of NK4 in the circulation and in multiple organs. Despite NK4 expression that was highest in the liver, toxicity in the liver was minimal. Ad.NK4‐mediated growth inhibition was associated with decreased blood vessel density and increased apoptosis in tumor tissues, which suggests that NK4 suppressed tumor growth as an angiogenesis inhibitor. Metastasis of B16F10 melanoma and Lewis lung carcinoma cells to the lung was potently inhibited by systemic Ad.NK4‐administration. Our results demonstrated that the adenovirus‐mediated induction of high levels of circulating NK4 significantly inhibited in vivo tumor growth and distant metastasis without obvious side effects. NK4 gene therapy is thus a safe and promising strategy for the treatment of cancer patients, and further validation in clinical trials is needed. (Cancer Sci 2009; 100: 1351–1358)
Involvement of G‐patch domain containing 2 overexpression in breast carcinogenesis
Authors: Lin Meng‐Lay , Fukukawa Chikako , Park Jae‐Hyun , Naito Kie , Kijima Kyoko , Shimo Arata , Ajiro Masahiko , Nishidate Toshihiko , Nakamura Yusuke , Katagiri Toyomasa
Published: April 2009

Through analysis of the detailed genome‐wide gene expression profiles of 81 breast tumors, we identified a novel gene, G‐patch domain containing 2 (GPATCH2), that was overexpressed in the great majority of breast cancer cases. Treatment of breast cancer cells MCF‐7 and T47D with siRNA against GPATCH2 effectively suppressed its expression, and resulted in the growth suppression of cancer cells, suggesting its essential role in breast cancer cell growth. We found an interaction of GPATCH2 protein with hPrp43, an RNA‐dependent ATPase. Their interaction could significantly enhance the ATPase activity of hPrp43 and induce a growth‐promoting effect on mammalian cells. Because northern blot analyses of normal human organs implied GPATCH2 to be a novel cancer/testis antigen, targeting GPATCH2 or inhibition of the interaction between GPATCH2 and hPrp43 could be a promising novel therapeutic strategy of breast cancer. (Cancer Sci 2009)
Evaluation of diquat against an acute experimental infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus (Rafinesque)
Authors: Darwish A M , Mitchell A J
Published: April 2009

A trial was performed to evaluate the efficacy of diquat (6,7‐dihydrodipyrido[1,2‐a:2′,1′‐c]pyrazinediium dibromide) against an acute experimental infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus. Diquat is an Environmental Protection Agency‐approved herbicide and has the potential to be legally and practically used against columnaris. Channel catfish were challenged, by cutaneous abrasion, and waterborne exposure to F. columnare and treated once at 22‐h post‐challenge with 2.5, 5.0, 10.0 and 15 mg L−1 of diquat active ingredient for 6 h. At the conclusion of the trial, 21‐day post‐challenge, diquat at 5.0, 10.0 and 15 mg L−1 significantly (P < 0.05) reduced the mortality of infected fish from 95% in the challenged non‐treated fish to 68%, 59% and 49%, respectively. In vitro, the minimum inhibitory concentration (MIC) of 23 isolates of F. columnare was assayed. The majority of the isolates had an MIC value of 5 μg mL−1 (15 of the 23 isolates). Infected fish exhibited acute clinical signs similar to a natural infection. The skin had severe ulcerative necrotizing dermatitis and the muscles had severe necrotizing myositis. The gills had severe multifocal necrotizing branchitis. The results demonstrate that diquat would reduce mortalities caused by an acute columnaris infection.
Detection and identification of aquatic mycobacteria in formalin‐fixed, paraffin‐embedded fish tissues
Authors: Pourahmad F , Thompson K D , Adams A , Richards R H
Published: April 2009

The isolation of mycobacteria from field samples is problematic, and isolation of the bacterium is sometimes not even attempted. The detection of mycobacteria through traditional histology using formalin‐fixed, paraffin‐embedded (FFPE) tissues is neither sensitive nor specific. However, detection of mycobacterial DNA from FFPE specimens, suspected of being infected with mammalian mycobacteriosis, is a routine clinical procedure. In the present study, a polymerase chain reaction (PCR)‐based method was used to detect and identify mycobacteria in FFPE specimens sampled from fish suspected of being infected with fish mycobacteriosis. A total of 45 fish tissue samples, comprising of 12 tissue samples obtained from experimentally infected fish and the remainder from fish naturally infected with mycobacteria, were analysed using a PCR protocol which amplifies a fragment of the mycobacterial 65 kDa heat‐shock protein (hsp65) gene. PCR‐restriction enzyme analysis and/or sequencing were employed to further analyse the PCR amplicons. The PCR results were compared with those obtained by histology and culture. Mycobacterial DNA was detected in 34 of the 45 samples examined, of which 16 samples (47%) showed granulomatous reactions on histological examination. Using histology as the gold standard, no false‐negative PCR results were obtained. Also, considering the presence or absence of granulomas as a diagnostic criterion, the sensitivity and specificity of PCR in 42 of the FFPE tissues were 16/16 (100%) and 8/26 (∼30.8%), respectively. Corresponding microbiological cultures were available for 15 cases, of which 13 were pure Mycobacterium cultures. Of these, 13 were PCR positive (100% sensitivity and 50% specificity). The PCR‐based methods used here proved sensitive, specific and rapid for the detection of mycobacteria in routinely processed paraffin wax‐embedded and formalin‐fixed histological samples, and the results of the study suggest that this method has potential use in retrospective epidemiological studies.
Experimental study of the susceptibility of Atlantic cod, Gadus morhua (L.), to infection with an IPNV strain pathogenic for Atlantic salmon, Salmo salar L
Authors: Urquhart K , Bowden T J , Buckett B‐E , Garcia J , Fryer R J , Ellis A E
Published: April 2009

Intraperitoneal (IP) injection, cohabitation and immersion routes of infection were used to determine if Atlantic cod, Gadus morhua (L.), of 1 and 3 g are susceptible to infectious pancreatic necrosis (IPN). Mortalities of cod injected IP were significantly higher when challenged with infectious pancreatic necrosis virus (IPNV) than with phosphate buffered saline. This is the first report of Atlantic cod mortalities caused by IPNV. Fish challenged by cohabitation had significantly higher mortalities than the controls, but mortalities of Atlantic cod challenged with IPNV by immersion were not significantly different from controls. Titres of IPNV in the tissues of infected fish were sometimes very high (range 102–1010 infectious units per gram of tissue) suggesting virus replication and titres of fish that died were generally higher than those of fish which survived. However, the relatively low mortality rates when challenged by cohabitation and immersion (20% and 17%, respectively), compared to the IP injection challenge (100%) suggest that 1 and 3 g cod have low susceptibility to IPN when challenged by more natural routes. These data strongly suggest that the cause of death of experimentally challenged cod was IPNV and further histological evidence for this came from 1 g cod challenged IP with IPNV in which the pancreas showed severe necrosis and heavy immunostaining for IPNV coincidentally with the peak of mortalities.
The Anorectic Effect of Neuropeptide AF is Associated with Satiety‐Related Hypothalamic Nuclei
Authors: Cline M. A. , Newmyer B. A. , Smith M. L.
Published: April 2009

Neuropeptide AF (NPAF), a member of the RFamide family, is encoded by the same gene as neuropeptide FF (NPFF), which causes short‐term anorexia. However, reports on the role of NPAF on appetite‐related process are lacking. Thus, i.c.v. injections of 4.0, 8.0 and 16.0 nmol NPAF were administered to chicks to observe its effect on food and water intake. Chicks treated with 8.0 and 16.0 nmol i.c.v. NPAF decreased both their food and water intake. Additionally, all doses of NPAF injected caused a similar reduction in whole blood glucose concentration 180 min after injection. In a second experiment, chicks that received i.c.v. NPAF had an increased number of c‐Fos immunoreactive cells in the dorsomedial, paraventricular (magnocellular and parvicellular parts) and ventromedial nuclei. The arcuate nucleus and lateral hypothalamic area were not affected. In a third experiment, NPAF‐treated chicks exhibited fewer feeding pecks and spent less time perching, whereas they spent an increased time in deep rest. Other behaviours, including exploratory pecking, escape attempts, defecations, distance moved, and time spent standing, sitting and preening, were not affected by NPAF injection. We conclude that NPAF causes anorectic effects that are associated with the hypothalamus.
Interleukin‐6 Gene Knockout Influences Energy Balance Regulating Peptides in the Hypothalamic Paraventricular and Supraoptic Nuclei
Authors: Benrick A. , Schéle E. , Pinnock S. B. , Wernstedt‐Asterholm I. , Dickson S. L. , Karlsson‐Lindahl L. , Jansson J.‐O.
Published: April 2009

Interleukin (IL)‐6 is a pro‐inflammatory cytokine that also affects metabolic function because IL‐6 depleted (IL‐6−/−) mice develop late‐onset obesity. IL‐6 appears to act in the central nervous system, presumably in the hypothalamus, to increase energy expenditure that appears to involve stimulation of the sympathetic nervous system. In the present study, we explored possible central mechanisms for the effects exerted by IL‐6 on body fat. Therefore, we measured the effects of IL‐6 depletion in IL‐6−/− mice on expression of key hypothalamic peptide genes involved in energy balance by the real time polymerase chain reaction. Additionally, co‐localisation between such peptides and IL‐6 receptor α was investigated by immunohistochemistry. IL‐6 deficiency decreased the expression of several peptides found in the paraventricular nucleus (PVN), which is a nucleus that has been attributed an adipostatic function. For example, corticotrophin‐releasing hormone (CRH), which is reported to stimulate the sympathetic nervous system, was decreased by 40% in older IL‐6−/− mice. Oxytocin, which is reported to prevent obesity, was also decreased in older IL‐6−/− animals, as was arginine vasopressin (AVP). The IL‐6 receptor α was abundantly expressed in the PVN, but also in the supraoptic nucleus, and was shown to be co‐expressed to a high extent with CRH, AVP, oxytocin and thyrotrophin‐releasing hormone. These data indicate that depletion of endogenous IL‐6, a body fat suppressing cytokine, is associated with the decreased expression of CRH and oxytocin (i.e. energy balance regulating peptides) as well as AVP in the PVN. Because IL‐6 receptor α is co‐expressed with CRH, oxytocin and AVP, IL‐6 could stimulate the expression of these peptides directly.
Ovarian‐Steroid Modulation of Locus Coeruleus Activity in Female Rats: Involvement in Luteinising Hormone Regulation
Authors: Szawka R. E. , Rodovalho G. V. , Monteiro P. M. , Carrer H. F. , Anselmo‐Franci J. A.
Published: April 2009

The noradrenergic nucleus locus coeruleus (LC) has been reported to regulate luteinising hormone (LH) secretion in female rats. Both oestrogen and progestin receptors have been demonstrated in LC neurones, suggesting that these cells are possibly responsive to variations in circulating levels of ovarian steroids. We therefore evaluated changes in the activity of LC neurones during the oestrous cycle and after ovarian‐steroid treatment in ovariectomised (OVX) rats, as determined by immunoreactivity to Fos‐related antigens (FRA), which comprises all of the known members of the Fos family. Effects of ovarian steroids on the firing rate of LC neurones were also determined in a slice preparation. The number of FRA/tyrosine hydroxylase (TH)‐immunoreactive (ir) neurones in the LC increased from 14.00–16.00 h on pro‐oestrus, coinciding with the onset of the LH surge and rise in plasma progesterone. FRA immunoreactivity was unaltered during dioestrus. Oestradiol‐treated OVX rats (OVX+E) displayed marked reduction in FRA/TH‐ir neurones in LC compared to oil‐treated OVX rats. Accordingly, oestradiol superfusion significantly reduced the spontaneous firing rate of LC neurones in slices from OVX rats. Compared to OVX+E, oestradiol‐treated rats injected with progesterone at 08.00 h (OVX+EP) exhibited higher number of FRA/TH‐ir neurones in the LC at 10.00 h and 16.00 h, and great amplification of the LH surge. Bath application of progesterone significantly increased the spontaneous firing rate of OVX+E LC neurones. Our data suggest that ovarian steroids may physiologically modulate the activity of LC neurones in females, with possible implications for LH secretion. Moreover, oestradiol and progesterone appear to exert opposite and complementary effects (i.e. whereas oestradiol inhibits, progesterone, after oestradiol priming, stimulates LC activity).
Biliary excretion of technetium‐99m‐sestamibi in wild‐type dogs and in dogs with intrinsic (ABCB1‐1Δ mutation) and extrinsic (ketoconazole treated) P‐glycoprotein deficiency
Authors: COELHO J. C. , TUCKER R. , MATTOON J. , ROBERTS G. , WAITING D. K. , MEALEY K. L.
Published: April 2009

P‐glycoprotein (P‐gp), the product of ABCB1 gene, is thought to play a role in the biliary excretion of a variety of drugs, but specific studies in dogs have not been performed. Because a number of endogenous (ABCB1 polymorphisms) and exogenous (pharmacological P‐gp inhibition) factors can interfere with normal P‐gp function, a better understanding of P‐gp’s role in biliary drug excretion is crucial in preventing adverse drug reactions and drug–drug interactions in dogs. The objectives of this study were to compare biliary excretion of technetium‐99m‐sestamibi (99mTc‐MIBI), a radio‐labelled P‐gp substrate, in wild‐type dogs (ABCB1 wild/wild), and dogs with intrinsic and extrinsic deficiencies in P‐gp function. Dogs with intrinsic P‐gp deficiency included ABCB1 mut/mut dogs, and dogs with presumed intermediate P‐gp phenotype (ABCB1 mut/wild). Dogs with extrinsic P‐gp deficiency were considered to be ABCB1 wild/wild dogs treated with the P‐gp inhibitor ketoconazole (5 mg/kg PO q12h × 9 doses). Results from this study indicate that ABCB1 mut/mut dogs have significantly decreased biliary excretion of99mTc‐MIBI compared with ABCB1 wild/wild dogs. Treatment with ketoconazole significantly decreased biliary excretion of99mTc‐MIBI in ABCB1 wild/wild dogs. P‐gp appears to play an important role in the biliary excretion of99mTc‐MIBI in dogs. It is likely that concurrent administration of a P‐gp inhibitor such as ketoconazole will decrease P‐gp‐mediated biliary excretion of other substrate drugs as well.
Depleted intestinal goblet cells and severe pathological changes in SCID mice infected with Heligmosomoides polygyrus
Authors: HASHIMOTO K. , UCHIKAWA R. , TEGOSHI T. , TAKEDA K. , YAMADA M. , ARIZONO N.
Published: April 2009

To determine the role of T cells and mast cells in intestinal pathology and immune expulsion of intestinal nematodes, worm burdens, goblet cell responses and villus structures were analysed in T‐ and B‐cell‐deficient severe combined immunodeficiency (SCID) mice, athymic nu/nu mice and mast cell deficient W/Wv mice after infection with the nematode Heligmosomoides polygyrus. SCID and nu/nu mice showed significantly higher worm burdens at week 9 post‐infection compared with the wild‐type controls. SCID and nu/nu mice showed compromised goblet cell hyperplasia and/or Muc 2 expression, indicating that both events are T‐cell dependant. On the other hand, the SCID mice showed increased pathology (villus atrophy and crypt hyperplasia) and increased numbers of proliferating cell nuclear antigen positive cells compared to the wild‐type controls. W/Wv mice, conversely, were able to expel the worms normally, had normal goblet cell hyperplasia, and did not demonstrate the changes in mucosal architecture seen in SCID mice, confirming that a normal mast cell response is not necessarily required for these changes. These results suggest that a functional T‐cell response, but not a mast cell response, is necessary for anti‐parasite responses, goblet cell function, and maintaining normal mucosal architecture.
Inhibitory role of toll‐like receptors agonists in Plasmodium yoelii liver stage development
Authors: CHEN J. , XU W. , ZHOU T. , DING Y. , DUAN J. , HUANG F.
Published: April 2009

It is well known that innate immune plays an important role in controlling the development of Plasmodium liver stage. However, little is known about the role of toll‐like receptors (TLR) signalling in the pre‐erythrocytic immunity against Plasmodium. Here, we found that pre‐treatment with individual TLR agonist pam3CSK4 (TLR2), poly(I:C) (TLR3), LPS (TLR4) and CpG (TLR9) could decrease significantly the liver malaria parasite load in mice for 58%, 63%, 75% and 88% respectively. Moreover, no parasitaemia was observed within 14 days in CpG group mice challenged with 100 sporozoites. At 24 h prior to CpG injection, administration of gadolinium chloride (GdCl3) led to the rebound of liver Plasmodium load through inhibiting selectively Kupffer cells (KC) phagocytosis capacity but failed to neutralize completely CpG‐induced immunity against malaria liver stage. Compared with the control, pre‐treatment of CpG up‐regulated hepatic pro‐inflammatory cytokines IL‐12, IFN‐γ and TNF‐α, but down‐regulated anti‐inflammatory cytokines IL‐10 and TGF‐β. Hence, our data demonstrated the inhibitory role of diverse TLR agonists in the Plasmodium development during pre‐erythrocytic stage. As the most robust agonist, CpG might inhibit the development of Plasmodium liver stage through regulation of intrahepatic inflammatory cytokines and enhancement of KC cells phagocytosis capacity.
Weed community characteristics and crop performance: a neighbourhood approach
Authors: POLLNAC F W , MAXWELL B D , MENALLED F D
Published: April 2009

The presence of weeds in crop fields often causes yield reductions. However, the effects of weed diversity have not been fully examined. This study tested the hypothesis that, holding density constant, increased weed species richness would decrease the effects of competition on spring wheat target plant performance. Measurements were taken from circular neighbourhoods (16.5 cm radius) with a single spring wheat plant surrounded by combinations of Setaria viridis, Avena fatua, Kochia scoparia, Thlaspi arvense and spring wheat plants, representing all combinations of neighbour species at four density levels. Using regression models, we found that species richness had no significant direct effect on spring wheat biomass, yield, or relative growth rate and that there were no significant neighbour species interaction terms. For weedy target plants, the presence of negative interaction terms suggested that increasing species richness had negative effects on growth of individual weed species. Additional analysis suggested that increased species richness may limit competitive ability of dominant weeds. Although we found no evidence of a direct effect of weed species richness on crop performance, increased weed species richness had no negative effect on spring wheat performance. Further, species richness of the weed community appears to influence weedy plant performance, which may offer a future opportunity to influence crop performance.
Demography of Salsola australis populations in the agricultural region of south‐west Australia
Authors: BORGER C P D , SCOTT J K , WALSH M , POWLES S B
Published: April 2009

Salsola australis is a widespread annual weed of broad‐scale cropping and pasture systems throughout Australia, and is of concern in the wheat belt of Western Australia. The population dynamics of S. australis was monitored at four sites in the Lake Grace district of Western Australia. At all sites, two to three cohorts emerged in summer and autumn or winter. While initial seedling density varied, density‐dependent mortality ensured that the total number of senesced plants at each site was not significantly different. Estimated seed production ranged from 95 to 19 596 seeds per plant, although seed viability was low (approximately 9% for seeds that were shed and 15% for seeds that were retained on the mature plants). Total annual seed production per unit area was highly variable within sites but was not significantly different between sites. These data will facilitate the development of improved control strategies for S. australis.
Assessment of management options for Salsola australis in south‐west Australia by transition matrix modelling
Authors: BORGER C P D , SCOTT J K , RENTON M , WALSH M , POWLES S B
Published: April 2009

A matrix model of the life cycle of Salsola australis was constructed, based on population ecology data collected from the district of Lake Grace, Western Australia. The model was used to assess potential control strategies for this summer annual weed within the Western Australian broad acre grain cropping system. The population growth rate (λ) of S. australis in the absence of weed control strategies was 1.49 and was virtually unaffected by the dormant seedbank. However, λ increased to 8.21 if it was assumed that a constant number of seed immigrated into the area in question from neighbouring populations of S. australis, through farm‐scale seed dispersal. As a result, effective weed management depended on reducing seed dispersal. The model determined that burning all senesced, mobile plants in late autumn, combined with herbicide control of the largest cohorts of S. australis in summer and autumn, reduced population growth rate to 0.79. This control strategy resulted in a 66.1% chance of the population becoming extinct over 25 years. Management strategies are proposed based on the results of the models and further research is required to validate their effectiveness and practicality in the field.
Discovery of a novel HLA‐Cw*08 allele, Cw*0817
Authors: Redman H. , Marlowe M. , Hanavan P. , Lake D. F. , Endres R. O.
Published: April 2009

This report describes the discovery and characterization of the HLA‐Cw*0817 allele.
Identification of B*9550, a novel B*15 allele, in a Mexican bone marrow donor from Veracruz State
Authors: Alaez C. , Munguía A. , Flores A. H. , Gomez K. , Gorodezky C.
Published: April 2009

Human leukocyte antigen‐B*9550 is a novel allele identified in a Mexican Mestizo bone marrow donor from Veracruz.
Resistance to broomrape in wild lentils (Lens spp.)
Authors: Fernández‐Aparicio M. , Sillero J. C. , Rubiales D.
Published: April 2009

Orobanche crenata, Orobanche aegyptiaca and Orobanche foetida constitute serious threats to grain legumes in the Mediterranean Basin. Lentil can be severely infected by O. crenata. It can also be damaged although with less virulence by O. aegyptiaca, and can only be slightly infected by O. foetida. Resistance breeding is hampered by scarcity of proper sources of resistance in cultivated lentil and of a reliable and practical screening procedure. A germplasm collection of 23 wild Lens spp. accessions was screened for resistance to O. crenata under field conditions. A wide range of responses was observed, from complete resistance to susceptibility. The higher levels of resistance were observed in accessions of Lens ervoides, Lens odemensis and Lens orientalis. Resistance of selected accessions proved to be mainly because of early hampered tubercle formation, with no observed necrosis of established tubercles. No differences were observed in levels of induction of broomrape germination, but necrosis of broomrape radicles was significant in some cases, particularly in the L. odemensis–O. foetida interaction. Escape because of the reduced root biomass diminishing the chance of contact between host and parasite was also observed in L. odemensis.
Ploidy manipulation in guinea grass (Panicum maximum Jacq., Poaceae) utilizing a Hybridization‐supplemented Apomixis‐components Partitioning Approach (HAPA)
Authors: Kaushal P. , Agrawal A. , Malaviya D. R. , Siddiqui S. A. , Roy A. K.
Published: April 2009

Ploidy manipulations are achieved by utilizing unreduced gametes, somatic chromosome doubling or haploidization. Apomixis, the asexual mode of reproduction through seeds, involves two of these phenomena viz., apomeiosis (unreduced embryo‐sac formation) and parthenogenesis (fertilization‐independent embryogenesis). The two components when partitioned may yield high frequencies of triploids and haploids, and thus, serve as a tool to manipulate ploidy levels when appropriately supplemented with hybridization schemes. Utilizing this Hybridization‐supplemented Apomixis‐components Partitioning Approach (HAPA), the largest ploidy series in a crop plant was generated in guinea grass (Panicum maximum Jacq.). Eighty‐nine diverse guinea grass accessions were characterized for their reproductive capacities to identify tetraploid (2n = 4x = 32) accession(s) with high apospory but reduced parthenogenetic capacity, thereby producing a high frequency of hexaploid seeds. Seeds from accession IG 04‐164 were utilized to produce a ploidy series by appropriately combining partitioned apomixis components and hybridizations in two seasonal cycles. We successfully obtained plants representing ploidies 3x, 4x, 5x, 6x, 8x and 9x, all derived from a single 4x progenitor. Production methodology, reproducibility and utilization of HAPA in cytogenetic and molecular studies are discussed.
Use of SSR markers to assess sexual vs. apomictic origin and ploidy level of breeding progeny derived from crosses of apple proliferation‐resistant Malus sieboldii and its hybrids with Malus × domestica cultivars
Authors: Bisognin C. , Seemüller E. , Citterio S. , Velasco R. , Grando M. S. , Jarausch W.
Published: April 2009

To obtain apple rootstocks resistant to apple proliferation and suitable to modern fruit growing, 24 cross‐combinations were performed over a 5‐year period using Malus sieboldii and its hybrids as donors of the resistance trait and standard apple rootstock Malus × domestica genotypes as donors of agronomic value. Breeding with these genotypes was achieved despite different degrees of apomixis and polyploidy. Sets of five to six locus‐specific microsatellite markers were identified for characterizing each progeny. Supported by flow cytometry these markers were applied to infer mode of reproduction, genomic constitution and ploidy level. Microsatellites allele composition identical to the maternal parent was revealed in 1668 of 3032 seedlings indicating seed formation through apomixis. Complete genetic recombination was found in 398 seedlings. The remaining hybrids displayed a higher ploidy than that of the parental plants which was consistent with the fertilization of unreduced egg cells. Thus, for each cross‐combination, microsatellite loci were identified which enabled a reliable prediction of the ploidy level. They can now be applied in routine screening to distinguish sexual from apomictic progeny.
A national cohort study of long‐course preoperative radiotherapy in primary fixed rectal cancer in Denmark
Authors: Bülow S. , Jensen L. H. , Altaf R. , Harling H. , Jensen M. , Laurberg S. , Lindegaard J. C. , Muhic A. , Vestermark L. ,   on behalf of the Danish Colorectal Cancer Group
Published: April 2009

Objective  Preoperative radiotherapy has been shown to enable a fixed rectal cancer to become resectable which in turn may result in long‐time survival. In this study, we analysed the outcome of long‐course preoperative radiotherapy in fixed rectal cancer in a national cohort including all Danish patients registered with primary inoperable rectal cancer and treated in the period May 2001 to December 2005.

Method  The study was based on surgical and demographic data from a continuously updated and validated national database. In addition, retrospective data were retrieved from all departments of radiotherapy concerning technique of radiotherapy, dose and fractionation and use of concomitant chemotherapy. Outcome was determined by actuarial analysis of local control, disease‐free survival and overall survival.

Results  A total of 258 patients with fixed rectal cancer received long‐course radiotherapy (> 45 Gy). The median age at diagnosis was 66 years (range: 32–85) and 185 (72%) patients were male. The resectability rate was 80%, and a R0 resection was obtained in 148 patients (57% of all patients and 61% of those operated). The 5‐year local recurrence rate for all patients was 5% (95% CI: 3–7%), and the actuarial distant recurrence rate was 41% (95% CI: 35–47%). The cumulative 5‐year disease‐free survival was 27% (95% CI: 22–32%) and overall 5‐year survival was 34% (95% CI: 29–39%).

Conclusions  This study is the first population‐based report on outcome of preoperative long‐course radiotherapy in a large unselected patient group with clinically fixed rectal cancer. Most patients could be resected with the intention of cure and one in three was alive after 5 years.
Diverting loop ileostomy after restorative proctocolectomy: predictors of poor outcome and poor quality of life
Authors: Scarpa M. , Ruffolo C. , Boetto R. , Pozza A. , Sadocchi L. , Angriman I.
Published: April 2009

Aim  Diverting loop ileostomy is used to minimize the impact of anastomotic complication after restorative proctocolectomy (RPC). However, the ileostomy itself may have complications and therefore affect quality of life (QOL). The aim of this study was to analyse the predictors of complications of the ileostomy formation and closure and of the QOL of these patients.

Method  Forty‐four consecutive patients who underwent RPC were enrolled. Records of the ileostomy follow‐up were retrieved from a prospectively collected database and QOL was assessed with the Stoma‐QOL questionnaire. Ileostomy site coordinates were measured. Univariate and multivariate analysis were performed.

Results  In this series, three patients experienced peristomal herniae, two ileostomy stenosis, seven ileostomy retraction and fourteen peristomal dermatitis. Emergency surgery was the only predictor of parastomal hernia (P = 0.017). Stenosis correlated with the distance from the umbilicus (τ = 0.24, P = 0.021). Use of standard rod and retraction were independent predictors of peristomal dermatitis (P = 0.049 and P = 0.001). Stoma‐QOL was directly correlated to the age of the patients and to the occurrence of parastomal hernia (P = 0.001 and P = 0.021, respectively). After stoma closure, two patients reported wound sepsis and seven suffered obstructive episodes.

Conclusion  The predictors of negative outcome after construction of a diverting loop ileostomy after RPC were urgent surgery, use of standard rod, the distance of the stoma site from the umbilicus, parastomal herniae and the older age of patients.
The number of lymph nodes in colon cancer specimens: what do the numbers really mean?
Authors: McPartland S. , Hyman N. , Blaszyk H. , Osler T.
Published: April 2009

Aim  Stage‐specific survival for colon cancer is improved when more lymph nodes are identified in the surgical specimen. This association is typically attributed to staging effect, but may instead be a surrogate for tumour biology.

Method  We retrospectively studied a cohort of 48 consecutively treated patients with Stage II colon cancer who underwent complete resection between January 2000 and December 2002. Archived H&E slides were reviewed for lymphocytic infiltration at the leading edge, presence and degree of sinus histiocytosis in the largest node and the presence of lymph node hyperplasia.

Results  The mean number of lymph nodes identified was 14.1 ± 9.4. T stage was strongly associated with the number of nodes identified (P = 0.01) and the presence of a significant degree of sinus histiocytosis approached statistical significance (P = 0.077). No statistically significant relationship existed between number of lymph nodes in a specimen and tumour location (P = 0.44), grade (P = 0.56) or lymphovascular invasion (P = 0.64).

Conclusions  T stage is highly associated with the number of nodes found in a colon cancer specimen; a significant degree of sinus histiocytosis may also be predictive. Finding more nodes may be a surrogate for tumour or host‐related factors that impact prognosis.
Economic evaluation of the treatment of grade II haemorrhoids: a comparison of stapled haemorrhoidopexy and rubber band ligation
Authors: McKenzie L. , De Verteuil R. , Cook J. , Shanmugam V. , Loudon M. , Watson A. J. M. , Vale L.
Published: April 2009

Objectives  Haemorrhoidal disease is a common condition causing considerable distress to individuals and significant cost to healthcare services. This paper explored the cost‐effectiveness of stapled haemorrhoidopexy (SH) compared with the non‐surgical intervention, rubber band ligation (RBL), for grade II symptomatic circumferential haemorrhoids.

Method  An economic evaluation alongside a randomized controlled trial conducted between October 2002 and February 2005. Adults were recruited and randomized to either SH or RBL. The same surgeon performed all procedures and investigators were blinded until analyses were completed. Primary outcomes measured at 52 weeks were cumulative costs to the NHS, clinical diagnosis of recurrence and quality adjusted life years (QALYs).

Results  Sixty symptomatic men and women with confirmed clinical diagnosis of grade II symptomatic haemorrhoids were randomized. Loss to follow‐up was up to 10% at 52 weeks. The mean cost for SH was greater than RBL (mean difference: £1483, 95% CI: 1339–1676); disease recurrence was lower (OR = 0.18, 95% CI: 0.03–0.86); and there was no evidence of a statistically significant difference in QALYs (−0.014, 95% CI: –0.076 to 0.051). SH was associated with a modest incremental cost per recurrence avoided at 12 months follow‐up (£4945). Based on current data, it was considered highly unlikely to be cost‐effective in terms of incremental cost per QALY.

Conclusions  There is insufficient evidence about the cost‐effectiveness of SH for grade II haemorrhoids to recommend its routine use in place of RBL. Further information is needed from larger trials with a longer‐term follow‐up to inform subsequent economic evaluation.
What is the natural history of internal rectal prolapse?
Authors: Wijffels N. A. , Collinson R. , Cunningham C. , Lindsey I.
Published: April 2009

Aim  The nature and clinical significance of internal rectal prolapse is controversial. Its natural history is unclear. Longitudinal cohort studies show rare progression to external prolapse but lack adequate follow‐up. We aimed to study the relationship of age to various stages of internal rectal prolapse using the Oxford Rectal Prolapse Grade (ORPG) and evaluate the influence of sex and vaginal delivery on this relationship.

Method  Internal rectal prolapsed (IRP) diagnosed at proctography and external rectal prolapse were graded using the ORPG. Age, sex and obstetric history were documented. Mean age of each prolapse grade (1–5) was analysed and regression analysis performed for age and prolapse. Subgroup analyses were made for males, and females with (V+) and without (V0) history of vaginal delivery.

Results  Sixty males (11%) and 471 females (89%) were studied. The difference in the mean ages of each group was statistically significant (grade 1,38.6; grade 2, 52.1; grade 3, 56.0; grade 4, 60.3 and grade 5, 66.5, P < 0.0001). On average male (8.7 years) and V0‐group (8.0 years) were younger than V+ group (95% CI difference 4.5–12.9 years, P < 0.0001, and 3.8–12.2 years, P < 0.0001, respectively). Males and V0‐group had weaker correlation between age and prolapse grade (r = 0.16 and r = 0.17, respectively, vs 0.41), and a faster prolapse progression rate than the V+ group.

Conclusion  These data demonstrate a strong relationship between age and prolapse grade, supporting the view of internal rectal prolapse as a precursor to external prolapse in the spectrum of rectal prolapse disease.
Single incision laparoscopic sigmoid colon resections without visible scar: a novel technique
Authors: Brunner W. , Schirnhofer J. , Waldstein‐Wartenberg N. , Frass R. , Weiss H.
Published: April 2009

Objective  On the way to ‘no‐scar’ techniques we developed a novel method for colorectal resection utilizing three intraumbilical trocars which results in a nonvisible postoperative scar.

Method  Two female patients (Age: 56a, 42a) underwent laparoscopic colorectal resection for diverticulitis and infiltrating endometriosis of the rectosigmoid colon, respectively. The entire operation was carried out transumbilically following the standardized principles of colorectal resection.

Results  The operative time was 110 and 180 min, respectively. No intraoperative adverse events or significant perioperative complication was noticed. The specimen measured 22 and 18 cm in length respectively. Estimated blood loss was minimal in both cases. Oral diet was resumed on postoperative day one. Patients were discharged on postoperative day 7 and day 6, respectively. At follow‐up, patients presented with an optimal cosmetic result without apparent scarring.

Conclusion  For the first time, a novel laparoscopic technique for sigmoid colon resection utilizing a single intraumbilical approach is presented. This new method allows further reduction of the surgical trauma and obviates any visible scar.