Protocol for the production of viable bimaternal mouse embryos

A reliable nuclear transfer method was first reported in 1983; it provided definite evidence that parthenogenetic embryos are lethal at early postimplantation in mammals. Subsequently, nuclear transfer has been extensively used as an important and versatile tool for investigating embryo and somatic-cell cloning and nucleo-cytoplasmic interactions. Further development of this technique has enabled the generation of bimaternal embryos containing two haploid sets of maternal genomes from female germ cells of different origins. By using a 2-d nuclear transfer system for oocyte reconstruction, viable mice can be produced solely from maternal genomes, without the participation of the paternal genome. This oocyte reconstruction system, as described in this protocol, could provide valuable guidelines for exploring the potential endowments of gametes and for conferring novel properties to them.     

Cartilage link protein interacts with neurocan, which shows hyaluronan binding characteristics different from CD44 and TSG-6

The interaction of neurocan with hyaluronan was qualitatively characterized with alkaline phosphatase fusion proteins secreted by mammalian cells. The wild type neurocan hyaluronan binding domain fused to alkaline phosphatase bound to immobilized hyaluronan under physiological as well as moderately hypertonic conditions, whereas its ability to bind to immobilized chondroitin sulfate dropped rapidly with increasing salt concentration. Strong hyaluronan binding ability was still evident when in both link modules within the hyaluronan binding domain a basic amino acid was mutated, which is well conserved among link modules of hyaluronan binding proteins. A strong enhancement of the binding of neurocan to immobilized hyaluronan was observed after preincubation of the immobilized hyaluronan with cartilage link protein. Moreover, this preincubation mediated also the binding of a fusion protein representing only the immunoglobulin module of neurocan linked to alkaline phosphatase, which showed no binding to immobilized hyaluronan alone. The interaction of the neurocan immunoglobulin module with link protein could also be shown by overlay blot analysis. These observations suggest that the hyaluronan binding characteristics of paired link modules are different from those of single link modules, and that the reported temporal co-expression of cartilage link protein and of neurocan in developing brain implicates the possibility of a cooperative function of these molecules.     

Knockout of the SREBP system increases production of the polyketide FR901512 in filamentous fungal sp. No. 14919 and lovastatin in Aspergillus terreus ATCC20542

Applied Microbiology and Biotechnology - In the production of useful microbial secondary metabolites, the breeding of strains is generally performed by random mutagenesis. However, because random...     

A case of infanticide in a recently found gelada population in Arsi, Ethiopia

There have been no reports of infanticide in wild gelada baboons and it has been argued that infanticide is not necessary in geladas, since the birth interval of female gelada can be shortened after takeover of a unit by a new leader male without infanticide. However, we observed an instance of infanticide in a newly-found wild gelada population in the Arsi Region of Ethiopia. After a leader male of the unit was severely wounded by a leopard attack, he was quite weakened. The second male of the unit, a young adult male, became the leader of the unit three weeks later, but the former leader continued to stay in the unit as a second male. After a week, two other adult males joined the unit which, therefore, came to include four adult males. The infanticide took place nine days later. The perpetrator was one of the immigrant males and he showed great interest in the mother of the unweaned victim infant. Although the perpetrator copulated with her after the infanticide, the usurper was found to own all three adult females after two weeks following the infanticide; i.e. the perpetrator could not own any female. The wounded former leader showed conspicuous protective behavior towards the victim's mother and the dead infant. One possible explanation for the occurrence of infanticide in this population of geladas is as follows. Gelada males in this area may be able to join units more easily to form multi-male units but then have shorter tenure in the units. Facing the unstable condition of units, they may sometimes engage in infanticide to increase their breeding opportunities, even before becoming a leader.     

Sociological and demographic characteristics of a recently found arsi gelada population in ethiopia

A new population of gelada baboons isolated from other known gelada populations living on the opposite side of the Rift Valley was found in 1989 in the Arsi region of Ethiopia (Mori &Belay, 1990). Subsequent observations were conducted on the new gelada population in three study periods of 16 to 55 days over three consecutive years, 1994–1996. When we compare this population with other well studied northern populations, densities were lower, band size was smaller and the ratio of juveniles to adult females was lower. Based onOhsawa andDunbar's (1984) criteria these results suggest that this population exploits the harshest environment among known gelada populations. This may be due to the severe effect of the dry season at low altitude in the study site. Band size was small in the study site. Association rates of units were low, and each unit behaved more independently than those of northern populations. Age-sex composition of units changed drastically between consecutive study periods, indicating unstable unit structure. Moreover, there seems to be a tendency in this population whereby several adult males easily invade and are incorporated into a unit but the unit is expected to thereafter divide. The phenomenon can be explained by two factors: (1) Each unit fed rather independently in this area, where units fed intensively on both plateau and the slope of the cliff; and (2) Males may be incorporated in a unit for defense against leopards. As the units were vary small in size and unstable as compared with the Semien gelada units, the traditional concept of a stable gelada unit formed by blood related females, does not seem to fit this population.     

Time-dependent attachment mechanism of bacterial pathogen during ice-ice infection in Kappaphycus alvarezii (Gigartinales, Rhodophyta)

The mechanism of infection by Vibrio sp. P11 promoting the ice-ice disease in Kappaphycus alvarezii was investigated in vitro. Its intensity of infection differs from that of another ice-ice promoter (Cytophaga sp. P25) by promoting the disease much faster. However, when secondary infection by other bacteria starts, its ability to compete with these bacteria gradually diminishes, whereas, infection by P25, although not displaying such drastic effects as P11, shows consistent competitive ability against other bacteria. Time-series infection experiments with application of polyclonal antibodies to specifically detect Vibrio sp. P11 revealed that this bacterium has a high affinity for the seaweed especially when the latter is stressed. It promotes the disease after a rapid increase in cell density of up to 10⁷ g⁻¹ (wet wt.) in the first 24 h. This bacterial cell build-up may take only 1–2 h on stressed thalli, but takes about 24 h on non-stressed thalli. Build-up is not sustainable in non-stressed thalli as high density is usually followed by a sudden decline in cell number believed to result from an algal defence against potential pathogens. Inoculation of the bacterium on thalli incubated in continuous culture system extends the time of bacterial attachment due to laminar flow and, possibly, competition by existing bacteria on the seaweed surface and in ambient seawater medium. Motility-driven cell attachment by this bacterium is suggested as an important factor for infection. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of lactoferrin and lactoperoxidase‐containing food on the oral microbiota of older individuals

The oral microbiota influences health and disease states. Some gram‐negative anaerobic bacteria play important roles in tissue destruction associated with periodontal disease. Lactoferrin (LF) and lactoperoxidase (LPO) are antimicrobial proteins found in saliva; however, their influence on the whole oral microbiota currently remains unknown. In this randomized, double‐blinded, placebo‐controlled study, the effects of long‐term ingestion of LF and LPO‐containing tablets on the microbiota of supragingival plaque and tongue coating were assessed. Forty‐six older individuals ingested placebo or test tablets after every meal for 8 weeks. The relative abundance of bacterial species was assessed by 16S rRNA gene high‐throughput sequencing. Most of the bacterial species in supragingival plaque and tongue coating that exhibited significant decreases in the test group were gram‐negative bacteria, including periodontal pathogens. Decreases in the total relative abundance of gram‐negative organisms in supragingival plaque and tongue coating correlated with improvements in assessed variables related to oral health, such as oral malodor and plaque accumulation. Furthermore, there was significantly less microbiota diversity in supragingival plaque at 8 weeks in the test group than in the placebo group and low microbiota diversity correlated with improvements in assessed variables related to oral health. These results suggest that LF and LPO‐containing tablets promote a shift from a highly diverse and gram‐negative‐dominated to a gram‐positive‐dominated community in the microbiota of supragingival plaque and tongue coating. This microbial shift may contribute to improvements in oral health, including oral malodor and state of the gingiva.

     

Further characterization of the PW peptide family that inhibits neuron differentiation in <Emphasis Type="Italic">Hydra</Emphasis>

From an evolutionary point of view, Hydra has one of the most primitive nervous systems among metazoans. Two different groups of peptides that affect neuron differentiation were identified in a systematic screening of peptide signaling molecules in Hydra. Within the first group of peptides, a neuropeptide, Hym-355, was previously shown to positively regulate neuron differentiation. The second group of peptides encompasses the PW family of peptides that negatively regulate neuron differentiation. In this study, we identified the gene encoding PW peptide preprohormone. Moreover, we made the antibody that specifically recognizes LPW. In situ hybridization and immunohistochemical analyses showed that the PW peptides and the gene encoding them were expressed in ectodermal epithelial cells throughout the body except for the basal disk. The PW peptides are produced by epithelial cells and are therefore termed “epitheliopeptides.” Together with Hym-355, the PW family peptides mediate communication between neurons and epithelial cells and thereby maintain a specific density of neurons in Hydra. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Toshio Takahashi, Osamu Koizumi equally contributed to this study.