Noxa, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis

A critical function of tumor suppressor p53 is the induction of apoptosis in cells exposed to noxious stresses. We report a previously unidentified pro-apoptotic gene, Noxa. Expression of Noxa induction in primary mouse cells exposed to x-ray irradiation was dependent on p53. Noxa encodes a Bcl-2 homology 3 (BH3)-only member of the Bcl-2 family of proteins; this member contains the BH3 region but not other BH domains. When ectopically expressed, Noxa underwent BH3 motif-dependent localization to mitochondria and interacted with anti-apoptotic Bcl-2 family members, resulting in the activation of caspase-9. We also demonstrate that blocking the endogenous Noxa induction results in the suppression of apoptosis. Noxa may thus represent a mediator of p53-dependent apoptosis.     

Production of trout offspring from triploid salmon parents

Many salmonids have become at risk of extinction. For teleosts whose eggs cannot be cryopreserved, developing techniques other than egg cryopreservation to save genetic resources is imperative. In this study, spermatogonia from rainbow trout were intraperitoneally transplanted into newly hatched sterile triploid masu salmon. Transplanted trout spermatogonia underwent spermatogenesis and oogenesis in male and female recipients, respectively. At 2 years after transplantation, triploid salmon recipients only produced trout sperm and eggs. With use of these salmon as parents, we successfully produced only donor-derived trout offspring. Thus, by transplanting cryopreserved spermatogonia into sterile xenogeneic recipients, we can generate individuals of a threatened species.     

Vesicle endocytosis requires dynamin-dependent GTP hydrolysis at a fast CNS synapse

Molecular dependence of vesicular endocytosis was investigated with capacitance measurements at the calyx of Held terminal in brainstem slices. Intraterminal loading of botulinum toxin E revealed that the rapid capacitance transient implicated as "kiss-and-run" was unrelated to transmitter release. The release-related capacitance change decayed with an endocytotic time constant of 10 to 25 seconds, depending on the magnitude of exocytosis. Presynaptic loading of the nonhydrolyzable guanosine 5'-triphosphate (GTP) analog GTPgS or dynamin-1 proline-rich domain peptide abolished endocytosis. These compounds had no immediate effect on exocytosis, but caused a use-dependent rundown of exocytosis. Thus, the guanosine triphosphatase dynamin-1 is indispensable for vesicle endocytosis at this fast central nervous system (CNS) synapse.     

Interplay of rotational, relaxational, and shear dynamics in solid ⁴He

Using a high-sensitivity torsional oscillator (TO) technique, we mapped the rotational and relaxational dynamics of solid helium-4 ((4)He) throughout the parameter range of the proposed supersolidity. We found evidence that the same microscopic excitations controlling the torsional oscillator motions are generated independently by thermal and mechanical stimulation. Moreover, a measure for the relaxation times of these excitations diverges smoothly without any indication for a critical temperature or critical velocity of a supersolid transition. Finally, we demonstrated that the combined temperature-velocity dependence of the TO response is indistinguishable from the combined temperature-strain dependence of the solid's shear modulus. This implies that the rotational responses of solid (4)He attributed to supersolidity are associated with generation of the same microscopic excitations as those produced by direct shear strain.     

DNA methylation analysis on satellite I region in blastocysts obtained from somatic cell cloned cattle

Many observations have been made on cloned embryos and on adult clones by somatic cell nuclear transfer (SCNT), but it is still unclear whether the progeny of cloned animals is presenting normal epigenetic status. Here, in order to accumulate the information for evaluating the normality of cloned cattle, we analyzed the DNA methylation status on satellite I region in blastocysts obtained from cloned cattle. Embryos were produced by artificial insemination (AI) to non‐cloned or cloned dams using semen from non‐cloned or cloned sires. After 7 days of AI, embryos at blastocyst stage were collected by uterine flushing. The DNA methylation levels in embryos obtained by using semen and/or oocytes from cloned cattle were similar to those in in vivo embryos from non‐cloned cattle. In contrast, the DNA methylation levels in SCNT embryos were significantly higher (P < 0.01) than those in in vivo embryos from non‐cloned and cloned cattle, approximately similar to those in somatic cells used as donor cells. Thus, this study provides useful information that epigenetic status may be normal in the progeny of cloned cattle, suggesting the normality of germline cells in cloned cattle.     

Mortierella wolfii keratomycosis in a horse

Purpose To describe a case of superficial keratomycosis caused by Mortierella wolfii (M. wolfii) in a horse. Methods A thoroughbred filly was presented with painful right eye of 2 days’ duration. A superficial corneal ulcer was observed ventrally together with multifocal punctuate opacities axially. Samples were collected by swabbing and scraping the ulcerated lesion and submitted for microbiologic and cytologic examination. Results Microscopic evaluation of debrided corneal tissue revealed the presence of nonseptate fungal hyphae, and culture of a corneal swab yielded fungal growth. Medical treatment with topical antifungal, antibiotic and autogenous serum and systemic anti‐inflammatory resolved the problem within 2 weeks. Conclusions Cytologic evaluation of a corneal scraping was useful to make a clinical diagnosis of keratomycosis. Based on the mycological characteristics, the fungus isolated from the corneal lesion was identified as M. wolfii. To the authors’ knowledge, this is the first case report of equine keratomycosis associated with this fungus, although the organism is known to infect various organs of cattle.     

Distribution of <Emphasis Type="Italic">tomato chlorotic dwarf viroid</Emphasis> in floral organs of tomato

In situ hybridization was used to analyze the distribution pattern of Tomato chlorotic dwarf viroid (TCDVd) in floral organs of tomato plants. Following TCDVd invasion of floral organs, it became localized only in sepals at an early developmental stage, then reached other floral organs at the flower opening stage, with the exception of part of the placenta and ovules. When distribution of TCDVd was compared with that of Potato spindle tuber viroid (PSTVd), TCDVd was not detected in the outer integument around the embryo sac even though PSTVd was able to invade there, suggesting that such specific distribution might reflect the frequent occurrence of viroid disease on crops caused by PSTVd-seed transmission.     

HPLC fluorometric determination of natural phytoplankton phycocyanin and its usefulness as cyanobacterial biomass in highly eutrophic shallow lake

A method for HPLC determination of phycocyanin in phytoplankton samples using gelchromatography with a fluorescence detector was developed to examine the use of phycocyanin as an index of the appearance and progress of cyanobacterial blooms in highly eutrophic lake. At least two types of phycocyanin with different molecular weights, each spectroscopically different from phycoerythrin and allophycocyanin, were found in natural phytoplankton samples. Changes in phycocyanin concentrations were clearly coupled to changes in chlorophyll-a concentrations during June to October while cyanobacterial blooms were occurring. The chlorophyll-a to phycocyanin relationship was linear at chlorophyll-a concentrations of less than 250 μg L^?1. The relationship between cyanobacterial carbon and phycocyanin concentration was also linear, suggesting that phycocyanin content may be a useful index of cyanobacterial biomass in highly eutrophic lakes where large cyanobacterial blooms occur.     

Effects of Ca2+ and sulfhydryl reductant on the polymerization of soybean glycinin catalyzed by mammalian and microbial transglutaminases

Two types of transglutaminases (TGases), Ca(2+)-dependent TGase derived from guinea pig liver (GTGase) and Ca(2+)-independent TGase derived from a variant of Streptoverticillium mobaraense (MTGase), were used to study the cross-linking of soybean 11S globulin (glycinin). The effects of sulfhydryl reductant (dithiothreitol, DTT) and Ca(2+) on the conformation and TGase-catalyzed polymerization of glycinin were investigated. The conformational change of glycinin was probed by spectral methods. The degree of cross-linking and the polymer (aggregate) formation were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and dynamic light scattering, respectively. Addition of DTT stimulated the TGase-catalyzed cross-linking reactions without destroying the secondary and tertiary structure of glycinin but did not influence the polymer or aggregate formation. It was found that Ca(2+) caused the formation of larger size polymers at lower concentrations, while it suppressed the polymerization at higher concentrations. In addition, the cross-linking behaviors of glycinin were shown to be different between MTGase- and GTGase-catalyzed systems.