Comparative genomics of mutualistic viruses of Glyptapanteles parasitic wasps

Comparative genome analysis of two endosymbiotic polydnaviruses from Glyptapanteles parasitic wasps reveals new insights into the evolutionary arms race between host and parasite

A founder MLH1 mutation in families from the districts of Modena and Reggio-Emilia in northern Italy with hereditary non-polyposis colorectal cancer associated with protein elongation and instability.

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Th Inducing POZ-Kruppel Factor (ThPOK) Is a Key Regulator of the Immune Response since the Early Steps of Colorectal Carcinogenesis

We purposed to evaluate the role of Th inducing POZ-Kruppel Factor (ThPOK), a transcriptional regulator of T cell fate, in tumour-induced immune system plasticity in colorectal carcinogenesis. The amounts of CD4+, CD8+ and CD56+ and ThPOK+ cells infiltrate in normal colorectal mucosa (NM), in dysplastic aberrant crypt foci (microadenomas, MA), the earliest detectable lesions in colorectal carcinogenesis, and in colorectal carcinomas (CRC), were measured, and the colocalization of ThPOK with the above-mentioned markers of immune cells was evaluated using confocal microscopy. Interestingly, ThPOK showed a prominent increase since MA. A strong colocalization of ThPOK with CD4 both in NM and in MA was observed, weaker in carcinomas. Surprisingly, there was a peak in the colocalization levels of ThPOK with CD8 in MA, which was evident, although to a lesser extent, in carcinomas, too. In conclusion, according to the data of the present study, ThPOK may be considered a central regulator of the earliest events in the immune system during colorectal cancer development, decreasing the immune response against cancer cells

Livestock-associated methicillin-resistant Staphylococcus aureus responsible for human colonization and infection in an area of Italy with high density of pig farming

BACKGROUND: Livestock-Associated MRSA (LA-MRSA) belonging to ST398 lineage, common among pigs and other animals, emerged in Central and Northern Europe, becoming a new risk factor for MRSA among farm workers. Strains belonging to ST398 can be responsible for human colonization and infection, mainly in areas with high livestock-farming. The aim of this study was to investigate the occurrence of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) human colonization and infections in an area of the Lombardy Region (Italy), the Italian region with the highest density of pig farming. METHODS: In the period March-April 2010, 879 nasal swabs were taken from subjects at admission to a local hospital serving an area of the Lombardy Region devoted to agriculture and farming. In the period March 2010-February 2011, all MRSA strains from community-acquired infection (CAI) observed in the same hospital, were collected. Molecular characterization of the isolates included SCCmec typing, spa typing and multilocus sequence typing (MLST). RESULTS: Out of 879 nasal swabs examined, 9 (1%) yielded MRSA. Five strains were assigned to sequence type (ST)398 (spa t899, 3 isolates; t108 and t2922, 1 isolate each) and were therefore categorized as LA-MRSA. The other 4 isolates were likely of hospital origin. No strains were positive for Panton-Valentine Leukocidin genes. Twenty MRSA isolates were detected from CAI, 17 were from skin and soft-tissue infections and 3 from other infections. An MRSA isolate from otitis externa was t899/ST398 and PVL-negative, hence categorized as LA-MRSA. Four isolates were assigned to t127/ST1. Eight strains were PVL-positive community acquired (CA)-MRSA and belonged to different clones, the most frequent being ST8. CONCLUSIONS: In an area of Italy with high density of pig farming, LA-MRSA is able to colonize the population and rarely to produce infections. Typical CA-MRSA is more common than LA-MRSA among CAI

Phlebotominae: vectors of leishmaniasis in the provinces of Santa Fe and Entre Ríos, Argentina

Fil: Salomón, Oscar D. ANLIS Dr.C.G.Malbrán. Centro Nacional de Diagnóstico e Investigación en Endemo-Epidemias; Argentina.Fil: Mocarbel, Nicolas J. Ministerio de Salud, Santa Fe; Argentina.Fil: Pedroni, Elena. Ministerio de Salud, Entre Rios; Argentina.Fil: Colombo, Javier. Ministerio de Salud, Santa Fe; Argentina.Fil: Sandillu, Monica. Ministerio de Salud, Entre Rios; Argentina.La transmisión de leishmaniasis tegumentaria (LT) se incrementó desde 1985 en 9 provincias argentinas. Santa Fe y Entre Ríos en dicho período no notificaron casos de transmisión autóctona comprobada, sin embargo en el año 2003 ocurrió un brote epidémico en Bella Vista, Corrientes, localidad que se encuentra en un área con continuidad ecológica y contigüidad geográfica con ambas provincias. Por ello, para determinar el riesgo potencial de transmisión de LT en las áreas próximas y al sur de Bella Vista se realizaron capturas de Phlebotominae en febrero del 2004, colectándose sobre las márgenes del río Paraná en Santa Fe (El Rabón, Villa Ocampo, Cayastá) y en Entre Ríos (La Paz, La Celina-Villa Urquiza) 860 ejemplares de Lutzomyia neivai (99.5%) y Lu. migonei (0.5 %), ambas especies con capacidad vectorial para Leishmania (V.) braziliensis. En Tartagal, Santa Fe, las capturas fueron consistentes con el paisaje de «chaco» residual: 7 ejemplares de Lu. nerivai, Lu. migonei y Lu. cortelezzii. Se destaca el riesgo potencial de transmisión epidémica de LT en estas provincias, especialmente por la tropicalización progresiva hacia el sur de la selva en galeria paranaense. Se recomiendan actividades de vigilancia clínica y vectorial. (EN) The transmission of tegumentary leishmaniasis (TL) has increased in 9 provinces of Argentina since 1985. Santa Fe and Entre Ríos did not record in this period autochtonous probed cases: however, an epidemic outbreak took place in 2003 in Bella Vista, Corrientes, located in an area with ecological continuity and contiguous to both provinces. In order to evaluate the potential risk of transmission of LT, Phlebotominae were captured at locations close to and southern from Bella Vista during February 2004. The traps located on the shores of Parana river in Santa Fe (El Rabón, Villa Ocampo, Cayastá), and Entre Ríos (La Paz. La Celina-Villa Urquiza) captured 860 individuals of Lutzomyia neivai (99.5%) and Lu. migonei (0.5 %), both species with vectorial capacity for Leishmania (V.) braziliensis. In Tartagal, Santa Fe, the captures were consistent with the residual «chaco» landscape, 7 individuals of Lu. nerivai, Lu. migonei and Lu. cortelezzii. The risk of LT epidemic transmission in these provinces is highlighted, mainly due to the progressive southern tropicalization of the paranaense gallery forest. Clinical and entomological surveillance is recommended

Genome-wide analysis of peptidase content and expression in a virulent and attenuated Babesia bovis strain pair

Bovipain-2 (in red), a Babesia bovis cysteine protease, is released into the host erythrocyte. Bovipain-2 is expressed in both virulent and attenuated B. bovis. DAPI stains the nuclei of two B. bovis. [Display omitted] ► Genome sequence of the virulent T2Bo strain reveals 66 peptidases which constitute 2% of the annotated coding sequences. ► Each of the peptidases identified in the virulent parent was represented in the attenuated daughter strain. ► Microarrays analyses of transcript levels for all 66 peptidase-encoding genes were investigated. ► Our study discusses our findings in regards to peptidase involvement in in vivo attenuation of B. bovis. Identifying virulence determinants in Apicomplexan parasites remains a major gap in knowledge for members within this phylum. We hypothesized that peptidases would segregate with virulence between a virulent parent Babesia bovis strain and an attenuated daughter strain derived by rapid in vivo passage. Using the complete genome sequence of the virulent T2Bo strain, 66 peptidases were identified and active sites confirmed. The presence, sequence identity and expression levels were tested for each of the 66 peptidases in the virulent parent and attenuated daughter T2Bo strains using whole genome, targeted sequencing approaches and microarrays analyses. Quantitative PCR revealed that there was no significant difference in peptidase expression between the virulent and attenuated strains. We conclude that while peptidases may well play a required role in B. bovis pathogenesis, neither loss of peptidase gene content nor reduced gene expression underlies the loss of virulence associated with in vivo passage and attenuation

Comparative transcriptome analysis of geographically distinct virulent and attenuated Babesia bovis strains reveals similar gene expression changes through attenuation

Background: Loss of virulence is a phenotypic adaptation commonly seen in prokaryotic and eukaryotic pathogens. This mechanism is not well studied, especially in organisms with multiple host and life cycle stages such as Babesia, a tick-transmitted hemoparasite of humans and animals. B. bovis, which infects cattle, has naturally occurring virulent strains that can be reliably attenuated in vivo. Previous studies suggest the virulence loss mechanism may involve post-genomic modification. We investigated the transcriptome profiles of two geographically distinct B. bovis virulent and attenuated strain pairs to better understand virulence loss and to gain insight into pathogen adaptation strategies. Results: Expression microarray and RNA-sequencing approaches were employed to compare transcriptome profiles of two B. bovis strain pairs, with each pair consisting of a virulent parental and its attenuated derivative strain. Differentially regulated transcripts were identified within each strain pair. These included genes encoding for VESA1, SmORFs, undefined membrane and hypothetical proteins. The majority of individual specific gene transcripts differentially regulated within a strain were not shared between the two strains. There was a disproportionately greater number of ves genes upregulated in the virulent parental strains. When compared with their attenuated derivatives, divergently oriented ves genes were included among the upregulated ves genes in the virulent strains, while none of the upregulated ves genes in the attenuated derivatives were oriented head to head. One gene family whose specific members were consistently and significantly upregulated in expression in both attenuated strains was spherical body protein (SBP) 2 encoding gene where SBP2 truncated copies 7, 9 and 11 transcripts were all upregulated. Conclusions: We conclude that ves heterodimer pair upregulation and overall higher frequency of ves gene expressions in the virulent strains is consistent with the involvement of this gene family in virulence. This is logical given the role of VESA1 proteins in cytoadherence of infected cells to endothelial cells. However, upregulation of some ves genes in the attenuated derivatives suggests that the consequence of upregulation is gene-specific. Furthermore, upregulation of the spherical body protein 2 gene family may play a role in the attenuated phenotype. Exactly how these two gene families may contribute to the loss or gain of virulence is discussed.EEA RafaelaFil: Pedroni, Monica J. Washington State University. College of Veterinary Medicine. Department of Veterinary Microbiology & Pathology. Program of Genomics; Estados UnidosFil: Sondgeroth, Kerry S. Washington State University. College of Veterinary Medicine. Department of Veterinary Microbiology & Pathology. Program of Genomics; Estados UnidosFil: Gallego-Lopez, Gina M. Washington State University. College of Veterinary Medicine. Department of Veterinary Microbiology & Pathology. Program of Genomics; Estados UnidosFil: Echaide, Ignacio Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Laboratorio de Inmunología y Parasitología; ArgentinaFil: Lau, Audrey OT. Washington State University. College of Veterinary Medicine. Department of Veterinary Microbiology & Pathology. Program of Genomics; Estados Unidos. Washington State University. College of Veterinary Medicine. Paul G. Allen School for Global Animal Health; Estados Unido

Characterization of acyl carrier protein and LytB in Babesia bovis apicoplast

Investigation of type II fatty acid and isoprenoid biosyntheses in Babesia resulted in the identification of two major components within the apicoplastic lumen. [Display omitted] ► This study illustrates a four membrane babesid apicoplast. ► Babesia bovis apicoplast resides adjacent to the nucleus. ► Acyl carrier protein and LytB are transcribed and translated in Babesia bovis. ► Isoprenoid biosynthesis likely exists in Babesia bovis. ► Type II fatty acid biosynthesis may not be present in Babesia bovis. The apicoplast is a highly specialized organelle that mediates required functions in the growth and replication of apicomplexan parasites. Despite structural conservation of the apicoplast among different parasite genera and species, there are also critical differences in the metabolic requirements of different parasites and at different stages of the life cycle. To specifically compare apicoplast pathways between parasites that have both common and unique stages, we characterized the apicoplast in Babesia bovis, which has only intraerythrocytic asexual stages in the mammalian host, and compared it to that of Plasmodium falciparum, which has both asexual intraerythrocytic and hepatic stages. Specifically focusing on the type II fatty acid (FASII) and isoprenoid (MEP) biosynthesis pathways, we searched for pathway components and retention of active sites within the genome, localized key components [acyl carrier protein (ACP) and 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (LytB)] to the apicoplast, and demonstrated that the N-terminal bipartite signals of both proteins are required and sufficient for trafficking to the apicoplast lumen. Using specific pharmacologic inhibition, we demonstrated that MEP biosynthesis may be disrupted and its presence is required for intraerythrocytic growth of B. bovis asexual stages, consistent with the genomic pathway analysis and with its requirement in the asexual erythrocytic stages of P. falciparum. In contrast, FASII biosynthesis may or may not be present and specific drug targets did not have any inhibitory effect to B. bovis intraerythrocytic growth, which is consistent with the lack of requirement for P. falciparum intraerythrocytic growth. However, genomic analysis revealed the loss of FASII pathway components in B. bovis whereas the pathway is intact for P. falciparum but regulated to be expressed when needed (hepatic stages) and silent when not (intraerythrocytic stages). The results indicate specialized molding of apicoplast biosynthetic pathways to meet the requirements of individual apicomplexan parasites and their unique intracellular niches

Genotypic diversity of merozoite surface antigen 1 of Babesia bovis within an endemic population

Investigation of the population dynamics of pathogens contributes to a better understanding of disease transmission. Herein, the extent of Babesia bovis genotypic strain diversity within a defined cohort is reported. Multiple genetically distinct strains of a pathogen circulate and compete for dominance within populations of animal reservoir hosts. Understanding the basis for genotypic strain structure is critical for predicting how pathogens respond to selective pressures and how shifts in pathogen population structure can lead to disease outbreaks. Evidence from related Apicomplexans such as Plasmodium, Toxoplasma, Cryptosporidium and Theileria suggests that various patterns of population dynamics exist, including but not limited to clonal, oligoclonal, panmictic and epidemic genotypic strain structures. In Babesia bovis, genetic diversity of variable merozoite surface antigen (VMSA) genes has been associated with disease outbreaks, including in previously vaccinated animals. However, the extent of VMSA diversity within a defined population in an endemic area has not been examined. We analyzed genotypic diversity and temporal change of MSA-1, a member of the VMSA family, in individual infected animals within a reservoir host population. Twenty-eight distinct MSA-1 genotypes were identified within the herd. All genotypically distinct MSA-1 sequences clustered into three groups based on sequence similarity. Two thirds of the animals tested changed their dominant MSA-1 genotypes during a 6-month period. Five animals within the population contained multiple genotypes. Interestingly, the predominant genotypes within those five animals also changed over the 6-month sampling period, suggesting ongoing transmission or emergence of variant MSA-1 genotypes within the herd. This study demonstrated an unexpected level of diversity for a single copy gene in a haploid genome, and illustrates the dynamic genotype structure of B. bovis within an individual animal in an endemic region. Co-infection with multiple diverse MSA-1 genotypes provides a basis for more extensive genotypic shifts that characterizes outbreak strains

Increased expression of cyclin-dependent kinase inhibitor 2 (CDKN2A) gene product P16INK4A in ovarian cancer is associated with progression and unfavourable prognosis

Paraffin sections from 190 epithelial ovarian tumours, including 159 malignant and 31 benign epithelial tumours, were analysed immunohistochemically for expression of cyclin-dependent kinase inhibitor 2 (CDKN2A) gene product p16INK4A (p16). Most benign tumours showed no p16 expression in the tumour cells, whereas only 11% of malignant cancers were p16 negative. A high proportion of p16-positive tumour cells was associated with advanced stage and grade, and with poor prognosis in cancer patients. For FIGO stage 1 tumours, a high proportion of p16-positive tumour cells was associated with poorer survival, suggesting that accumulation of p16 is an early event of ovarian tumorigenesis. In contrast to tumour cells, high expression of p16 in the surrounding stromal cells was not associated with the stage and grade, but was associated with longer survival. When all parameters were combined in multivariate analysis, high p16 expression in stromal cells was not an independent predictor for survival, indicating that low p16 expression in stromal cells is associated with other markers of tumour progression. High expression of p16 survival in the stromal cells of tumours from long-term survivors suggests that tumour growth is limited to some extent by factors associated with p16 expression in the matrix