Characterization of a Strain of Mycoplasma hominis Lacking 120 kDa Membrane Protein Isolated from Vero Cell Culture

A strain of Mycoplasma hominis lacking a major membrane protein of 120 kDa was isolated from a Vero cell culture. This strain showed very slow growth rate and formed nipple-less colonies on agar medium.     

Serologic Relationship of Coxsackie A16 Viruses from the Epidemic of Hand-Foot-and-Mouth Disease in Japan, 1970, to the Prototype Strain

In 1970, a great outbreak of hand-foot-and-mouth disease (HFMD) due to Coxsackie A16 virus (Cox A16) occurred throughout Japan. When serologic relationship between the viruses isolated during the epidemic and the prototype strain of the serotype was examined by the tube neutralization test, the crude new isolates were found to be poorly neutralized by both an antiserum against the prototype strain and those against the isolates, although they were neutralized significantly in the plaque reduction test. However, about 2% of virus in crude suspensions of the isolates remained unneutralized even in the plaque reduction test and this fraction could be eliminated by filtering the virus materials through a 100 mμ Millipore filter. Therefore, the difficulty in neutralization of the isolates by the tube method could be accounted for by the presence of aggregated viruses. Even when filtered viruses were used, the reciprocal neutralization kinetic studies revealed a significant serological difference between the isolates and the prototype strain. Such serological properties of the isolates were not influenced by the cell types used for virus isolation or passages. All the results suggest that the Cox A16 isolates from the epidemic of HFMD in Japan, 1970, are serologically different from the prototype strain.     

Establishment and proteomic characterization of a novel cell line,NCC-UPS2-C1, derived from a patient with undifferentiated pleomorphic sarcoma

Undifferentiated pleomorphic sarcoma (UPS) is an aggressive mesenchymal malignancy requiring novel therapeutic approaches to improve clinical outcome. Patient-derived cancer cell lines are an essential tool for investigating molecular mechanisms underlying cancer initiation and development; however, there is a lack of patient-derived cell lines of UPS available for research. The objective of this study was to develop a patient-derived cell model of UPS. A cell line designated NCC-UPS2-C1 was established from the primary tumor tissue of an 84-yr-old female patient with UPS. The short tandem repeat pattern of NCC-UPS2-C1 cells was identical to that of the original tumor and distinct from that of any other cell lines deposited in public cell banks. NCC-UPS2-C1 cells were maintained as a monolayer culture for over 80 passages during 30 mo and exhibited spindle-like morphology, continuous growth, and ability for spheroid formation and invasion. Proteomic profiling using mass spectrometry and functional treemap analysis revealed that the original tumor and the derived NCC-UPS2-C1 cells had similar but distinct protein expression patterns. Our results indicate that a novel UPS cell line was successfully established and could be used to study UPS development and effects of anti-cancer drugs. However, the revealed difference between proteomes of the original tumor and NCC-UPS2-C1 cells should be further investigated to determine the appropriate applications of this cell line in UPS research.     

Atomic structure of the clamp loader small subunit from Pyrococcus furiosus.

In eukaryotic DNA replication, replication factor-C (RFC) acts as the clamp loader, which correctly installs the sliding clamp onto DNA strands at replication forks. The eukaryotic RFC is a complex consisting of one large and four small subunits. We have determined the crystal structure of the clamp loader small subunit (RFCS) from Pyrococcus furiosus. The six subunits, of which four bind ADP in their canonical nucleotide binding clefts, assemble into a dimer of semicircular trimers. The crescent-like architecture of each subunit formed by the three domains resembles that of the delta' subunit of the E. coli clamp loader. The trimeric architecture of archaeal RFCS, with its mobile N-terminal domains, involves intersubunit interactions that may be conserved in eukaryotic functional complexes.     

Effects of plant hybridization on the structure and composition of a highly rich community of cynipid gall wasps: the case of the oak hybrid complex <Emphasis Type="Italic">Quercus magnoliifolia</Emphasis> x <Emphasis Type="Italic">Quercus resinosa</Emphasis> in Mexico

The richness and composition of herbivore communities can be influenced by the genetic variation of host plants. Hybrid plant populations are ideal to test these effects because they usually harbor high genetic variation and display a mosaic of phenotypic characters. The goal of this study was to examine the effect of hybridization between two Mexican white oaks, Q. magnoliifolia and Q. resinosa, on the composition and diversity of the associated cynipid gall wasp community. We used eight nuclear microsatellite markers to genotype 150 oak individuals sampled at three different altitudes at the Tequila volcano and conducted monthly samplings of galls in each individual over the course of 2 years. A Bayesian assignment analysis indicated genetic admixture between the two oak species at the study site and allowed classifying individuals as Q. magnoliifolia, Q. resinosa or hybrids. Gall morphospecies richness was significantly higher in the hybrids, intermediate in Q. magnoliifolia and lower in Q. resinosa. Overall, 48 different gall morphospecies were found, with 21 of them being shared among the three groups of plants, 13 between two groups of plants, and 14 were unique to one group of plants, with eight of these being found in hybrids. Several of the shared galls showed differences in abundance among plant groups. Therefore, genetic structure in this oak complex significantly influences the diversity and composition of the associated gall wasp community, and hybrid individuals are probably acting as potential sinks and bridges for the colonization of plant hosts by these highly specialized insect species.     

Scaffold Protein Ahk1, Which Associates with Hkr1, Sho1, Ste11, and Pbs2, Inhibits Cross Talk Signaling from the Hkr1 Osmosensor to the Kss1 Mitogen-Activated Protein Kinase

In the budding yeast Saccharomyces cerevisiae, osmostress activates the Hog1 mitogen-activated protein kinase (MAPK), which regulates diverse osmoadaptive responses. Hkr1 is a large, highly glycosylated, single-path transmembrane protein that is a putative osmosensor in one of the Hog1 upstream pathways termed the HKR1 subbranch. The extracellular region of Hkr1 contains both a positive and a negative regulatory domain. However, the function of the cytoplasmic domain of Hkr1 (Hkr1-cyto) is unknown. Here, using a mass spectrometric method, we identified a protein, termed Ahk1 (Associated with Hkr1), that binds to Hkr1-cyto. Deletion of the AHK1 gene (in the absence of other Hog1 upstream branches) only partially inhibited osmostress-induced Hog1 activation. In contrast, Hog1 could not be activated by constitutively active mutants of the Hog1 pathway signaling molecules Opy2 or Ste50 in ahk1Δ cells, whereas robust Hog1 activation occurred in AHK1⁺ cells. In addition to Hkr1-cyto binding, Ahk1 also bound to other signaling molecules in the HKR1 subbranch, including Sho1, Ste11, and Pbs2. Although osmotic stimulation of Hkr1 does not activate the Kss1 MAPK, deletion of AHK1 allowed Hkr1 to activate Kss1 by cross talk. Thus, Ahk1 is a scaffold protein in the HKR1 subbranch and prevents incorrect signal flow from Hkr1 to Kss1.