Asynchrony between pronuclear development and protein synthetic changes in zygotes of the mouse derived from oocytes aged postovulation in vivo and fertilized in vivo

The pattern of proteins synthesized by one-cell embryos derived from unaged oocytes and oocytes aged postovulation in vivo was analyzed by means of 35S-methionine labeling and gel electrophoresis. The oocytes were obtained after ovulation induction by an injection of luteinizing hormone-releasing hormone (LHRH) at proestrus or after a superovulation procedure. The analysis was performed in unfertilized aged and unaged secondary oocytes and in zygotes derived from them. The patterns of proteins synthesized by secondary oocytes from all experimental groups were very similar: The oocytes showed a predominant synthesis of 35 kDa proteins. Zygotes from aged as well as unaged LHRH-induced oocytes also showed a predominant synthesis of one group of polypeptides with a relative molecular weight of about 35 kDa. The proteins of the 35 kDa protein complex migrated in an upper (u), middle (m), or lower (l) band in 10% polyacrylamide sodium dodecyl sulfate (SDS) gels. The u- and m-band 35 kDa proteins were shown to be synthesized by secondary oocytes. Early pronuclear zygotes from unaged LHRH-induced oocytes synthesized u- and m- but no l-band 35 kDa proteins. In contrast, part (38%, n = 47) of the early pronuclear zygotes from aged LHRH-induced oocytes did synthesize the l-band 35 kDa proteins. Late pronuclear zygotes (LPZ) from aged as well as unaged oocytes synthesized predominantly the l-band 35 kDa proteins. However, although only 5.8% (n = 51) of LPZ from unaged oocytes synthesize m- and l-band 35 kDa proteins, these bands of proteins are present in 25% (n = 24) of the LPZ from aged oocytes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Development of enterobacterium-specific oligonucleotide probes based on the surface-exposed regions of outer membrane proteins

Outer membrane proteins of members of the family Enterobacteriaceae consist of conserved membrane-spanning segments and hypervariable, surface-exposed regions. We demonstrate that the hypervariable DNA segments corresponding to the surface-exposed regions of these proteins can be used to develop specific DNA probes for the identification of members of the family Enterobacteriaceae.     

Feed Conversion,Survival and Development,and Composition of Four Insect Species on Diets Composed of Food By-Products

A large part of the environmental impact of animal production systems is due to the production of feed. Insects are suggested to efficiently convert feed to body mass and might therefore form a more sustainable food and/or feed source. Four diets were composed from by-products of food manufacturing and formulated such as to vary in protein and fat content. These were offered to newly hatched Argentinean cockroaches, black soldier flies, yellow mealworms, and house crickets. The first two species are potentially interesting as a feed ingredient, while the latter two are considered edible for humans. Feed conversion efficiency, survival, development time, as well as chemical composition (nitrogen, phosphorus, and fatty acids), were determined. The Argentinean cockroaches and the black soldier flies converted feed more efficiently than yellow mealworms, and house crickets. The first two were also more efficient than conventional production animals. On three of the four diets yellow mealworms and house crickets had a feed conversion efficiency similar to pigs. Furthermore, on the most suitable diet, they converted their feed as efficiently as poultry, when corrected for edible portion. All four species had a higher nitrogen-efficiency than conventional production animals, when corrected for edible portion. Offering carrots to yellow mealworms increased dry matter- and nitrogen-efficiency and decreased development time. Diet affected survival in all species but black soldier flies, and development time was strongly influenced in all four species. The chemical composition of Argentinean cockroaches was highly variable between diets, for black soldier flies it remained similar. The investigated species can be considered efficient production animals when suitable diets are provided. Hence, they could form a sustainable alternative to conventional production animals as a source of feed or food.     

Copy number alterations and allelic ratio in relation to recurrence of rectal cancer

Background

In rectal cancer, total mesorectal excision surgery combined with preoperative (chemo)radiotherapy reduces local recurrence rates but does not improve overall patient survival, a result that may be due to the harmful side effects and/or co-morbidity of preoperative treatment. New biomarkers are needed to facilitate identification of rectal cancer patients at high risk for local recurrent disease. This would allow for preoperative (chemo)radiotherapy to be restricted to high-risk patients, thereby reducing overtreatment and allowing personalized treatment protocols. We analyzed genome-wide DNA copy number (CN) and allelic alterations in 112 tumors from preoperatively untreated rectal cancer patients. Sixty-six patients with local and/or distant recurrent disease were compared to matched controls without recurrence. Results were validated in a second cohort of tumors from 95 matched rectal cancer patients. Additionally, we performed a meta-analysis that included 42 studies reporting on CN alterations in colorectal cancer and compared results to our own data.

Results

The genomic profiles in our study were comparable to other rectal cancer studies. Results of the meta-analysis supported the hypothesis that colon cancer and rectal cancer may be distinct disease entities. In our discovery patient study cohort, allelic retention of chromosome 7 was significantly associated with local recurrent disease. Data from the validation cohort were supportive, albeit not statistically significant, of this finding.

Conclusions

We showed that retention of heterozygosity on chromosome 7 may be associated with local recurrence in rectal cancer. Further research is warranted to elucidate the mechanisms and effect of retention of chromosome 7 on the development of local recurrent disease in rectal cancer.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1550-0) contains supplementary material, which is available to authorized users.     

FGF signalling through RAS/MAPK and PI3K pathways regulates cell movement and gene expression in the chicken primitive streak without affecting E-cadherin expression

Background  

FGF signalling regulates numerous aspects of early embryo development. During gastrulation in amniotes, epiblast cells undergo an epithelial to mesenchymal transition (EMT) in the primitive streak to form the mesoderm and endoderm. In mice lacking FGFR1, epiblast cells in the primitive streak fail to downregulate E-cadherin and undergo EMT, and cell migration is inhibited. This study investigated how FGF signalling regulates cell movement and gene expression in the primitive streak of chicken embryos.     

Gastric cancers of Western European and African patients show different patterns of genomic instability

Background

Infection with H. pylori is important in the etiology of gastric cancer. Gastric cancer is infrequent in Africa, despite high frequencies of H. pylori infection, referred to as the African enigma. Variation in environmental and host factors influencing gastric cancer risk between different populations have been reported but little is known about the biological differences between gastric cancers from different geographic locations. We aim to study genomic instability patterns of gastric cancers obtained from patients from United Kingdom (UK) and South Africa (SA), in an attempt to support the African enigma hypothesis at the biological level.

Methods

DNA was isolated from 67 gastric adenocarcinomas, 33 UK patients, 9 Caucasian SA patients and 25 native SA patients. Microsatellite instability and chromosomal instability were analyzed by PCR and microarray comparative genomic hybridization, respectively. Data was analyzed by supervised univariate and multivariate analyses as well as unsupervised hierarchical cluster analysis.

Results

Tumors from Caucasian and native SA patients showed significantly more microsatellite instable tumors (p < 0.05). For the microsatellite stable tumors, geographical origin of the patients correlated with cluster membership, derived from unsupervised hierarchical cluster analysis (p = 0.001). Several chromosomal alterations showed significantly different frequencies in tumors from UK patients and native SA patients, but not between UK and Caucasian SA patients and between native and Caucasian SA patients.

Conclusions

Gastric cancers from SA and UK patients show differences in genetic instability patterns, indicating possible different biological mechanisms in patients from different geographical origin. This is of future clinical relevance for stratification of gastric cancer therapy.

     

Assessing non-target effects and host feeding of the exotic parasitoid <Emphasis Type="Italic">Apanteles taragamae</Emphasis>, a potential biological control agent of the cowpea pod borer <Emphasis Type="Italic">Maruca vitrata</Emphasis>

Apanteles taragamae Viereck is a larval parasitoid introduced in Benin for classical biological control of the cowpea pod borer Maruca vitrata Fabricius. In the laboratory, we evaluated the effects of A. taragamae on non-target herbivore species, and on another parasitoid of M. vitrata, i.e. the egg-larval parasitoid Phanerotoma leucobasis Kriechbaumer. Furthermore, we addressed the host feeding behaviour of A. taragamae. The host specificity of A. taragamae was assessed by offering six other lepidopteran species to the wasp. The competitive ability of A. taragamae was studied by providing the wasp with one- and two-days-old M. vitrata larvae that had hatched from eggs previously parasitized by P. leucobasis. Controls consisted of eggs and larvae offered only to P. leucobasis and A. taragamae, respectively. None of the other six lepidopteran species was successfully parasitized by A. taragamae. The larval parasitoid A. taragamae outcompeted the egg-larval parasitoid P. leucobasis when offered two-days-old host larvae. Competition between the two parasitoid species did not significantly affect one-day-old host larvae that were less suitable to A. taragamae. Host feeding by A. taragamae did not affect survival of one-day-old or two-days-old M. vitrata larvae. However, the percentage parasitism of two-days-old larvae was significantly reduced when exposed to female A. taragamae wasps that had been starved during 48 h. The data are discussed with regard to host specificity, host feeding patterns and to factors underlying the outcome of intrinsic competition between parasitoid species.     

Intimal lining layer macrophages but not synovial sublining macrophages display an IL-10 polarized-like phenotype in chronic synovitis

Introduction

Synovial tissue macrophages play a key role in chronic inflammatory arthritis, but the contribution of different macrophage subsets in this process remains largely unknown. The main in vitro polarized macrophage subsets are classically (M1) and alternatively (M2) activated macrophages, the latter comprising interleukin (IL)-4 and IL-10 polarized cells. Here, we aimed to evaluate the polarization status of synovial macrophages in spondyloarthritis (SpA) and rheumatoid arthritis (RA).

Methods

Expression of polarization markers on synovial macrophages, peripheral blood monocytes, and in vitro polarized monocyte-derived macrophages from SpA versus RA patients was assessed by immunohistochemistry and flow cytometry, respectively. The polarization status of the intimal lining layer and the synovial sublining macrophages was assessed by double immunofluorescence staining.

Results

The expression of the IL-10 polarization marker cluster of differentiation 163 (CD163) was increased in SpA compared with RA intimal lining layer, but no differences were found in other M1 and M2 markers between the diseases. Furthermore, no significant phenotypic differences in monocytes and in vitro polarized monocyte-derived macrophages were seen between SpA, RA, and healthy controls, indicating that the differential CD163 expression does not reflect a preferential M2 polarization in SpA. More detailed analysis of intimal lining layer macrophages revealed a strong co-expression of the IL-10 polarization markers CD163 and cluster of differentiation 32 (CD32) but not any of the other markers in both SpA and RA. In contrast, synovial sublining macrophages had a more heterogeneous phenotype, with a majority of cells co-expressing M1 and M2 markers.

Conclusions

The intimal lining layer but not synovial sublining macrophages display an IL-10 polarized-like phenotype, with increased CD163 expression in SpA versus RA synovitis. These differences in the distribution of the polarized macrophage subset may contribute to the outcome of chronic synovitis.     

Effect of Maruca vitrata (Lepidoptera: Crambidae) host plants on life‐history parameters of the parasitoid Apanteles taragamae (Hymenoptera: Braconidae)

Abstract The effect of four host plant species of the herbivore Maruca vitrata Fabricius (Lepidoptera: Crambidae) on development time, longevity, fecundity and sex ratio of the parasitoid Apanteles taragamae Viereck (Hymenoptera: Braconidae) was investigated under laboratory conditions. The larvae were parasitized when in the second instar. Maruca vitrata larvae were fed with flowers of four legumes, that is, Vigna unguiculata (cowpea), Sesbania rostrata, Lonchocarpus sericeus and Pterocarpus santalinoides, or an artificial diet both before and after parasitization. The parasitoid did not develop in hosts feeding on L. sericeus or V. unguiculata at 25°C, or on P. santalinoides at 25°C or 29°C. Apanteles taragamae had the shortest development time on artificial diet at both 25°C and 29°C while the longest development time was recorded on L. sericeus at 29°C. Female wasps took longer to develop compared to males at the two temperatures, regardless of the feeding substrate of their host. The longevity of the wasps at 25°C varied among feeding substrates, but not at 29°C. Survival rate of parasitized larvae depends on the feeding substrate. Moreover, infection of host larvae with Maruca vitrata multi‐nucleopolyhedrovirus (MaviMNPV) killed larger proportions of parasitized larvae at 25°C than at 29°C, which was likely caused by the difference in parasitoid developmental rate. The proportion of female parasitoids was lowest on L. sericeus. The daily fecundity showed a nonlinear trend regardless of the feeding substrate, indicating that A. taragamae is a pro‐ovigenic species. The data support the slow growth–high mortality hypothesis.