Methane and soil CO2 production from current‐season photosynthates in a rice paddy exposed to elevated CO2 concentration and soil temperature

Quantification of rhizodeposition (root exudates and root turnover) represents a major challenge for understanding the links between above‐ground assimilation and below‐ground anoxic decomposition of organic carbon in rice paddy ecosystems. Free‐air CO₂ enrichment (FACE) fumigating depleted ¹³CO₂ in rice paddy resulted in a smaller ¹³C/¹²C ratio in plant‐assimilated carbon, providing a unique measure by which we partitioned the sources of decomposed gases (CO₂ and CH₄) into current‐season photosynthates (new C) and soil organic matter (old C). In addition, we imposed a soil‐warming treatment nested within the CO₂ treatments to assess whether the carbon source was sensitive to warming. Compared with the ambient CO₂ treatment, the FACE treatment decreased the ¹³C/¹²C ratio not only in the rice‐plant carbon but also in the soil CO₂ and CH₄. The estimated new C contribution to dissolved CO₂ was minor (ca. 20%) at the tillering stage, increased with rice growth and was about 50% from the panicle‐formation stage onwards. For CH₄, the contribution of new C was greater than for heterotrophic CO₂ production; ca. 40–60% of season‐total CH₄ production originated from new C with a tendency toward even larger new C contribution with soil warming, presumably because enhanced root decay provided substrates for greater CH₄ production. The results suggest a fast and close coupling between photosynthesis and anoxic decomposition in soil, and further indicate a positive feedback of global warming by enhanced CH₄ emission through greater rhizodeposition.     

Cytoplasmic Location of α1A Voltage-Gated Calcium Channel C-Terminal Fragment (Cav2.1-CTF) Aggregate Is Sufficient to Cause Cell Death

The human α_1A voltage-dependent calcium channel (Ca_v2.1) is a pore-forming essential subunit embedded in the plasma membrane. Its cytoplasmic carboxyl(C)-tail contains a small poly-glutamine (Q) tract, whose length is normally 4∼19 Q, but when expanded up to 20∼33Q, the tract causes an autosomal-dominant neurodegenerative disorder, spinocerebellar ataxia type 6 (SCA6). A recent study has shown that a 75-kDa C-terminal fragment (CTF) containing the polyQ tract remains soluble in normal brains, but becomes insoluble mainly in the cytoplasm with additional localization to the nuclei of human SCA6 Purkinje cells. However, the mechanism by which the CTF aggregation leads to neurodegeneration is completely elusive, particularly whether the CTF exerts more toxicity in the nucleus or in the cytoplasm. We tagged recombinant (r)CTF with either nuclear-localization or nuclear-export signal, created doxycyclin-inducible rat pheochromocytoma (PC12) cell lines, and found that the CTF is more toxic in the cytoplasm than in the nucleus, the observations being more obvious with Q28 (disease range) than with Q13 (normal-length). Surprisingly, the CTF aggregates co-localized both with cAMP response element-binding protein (CREB) and phosphorylated-CREB (p-CREB) in the cytoplasm, and Western blot analysis showed that the quantity of CREB and p-CREB were both decreased in the nucleus when the rCTF formed aggregates in the cytoplasm. In human brains, polyQ aggregates also co-localized with CREB in the cytoplasm of SCA6 Purkinje cells, but not in other conditions. Collectively, the cytoplasmic Ca_v2.1-CTF aggregates are sufficient to cause cell death, and one of the pathogenic mechanisms may be abnormal CREB trafficking in the cytoplasm and reduced CREB and p-CREB levels in the nuclei.     

Identification of epidermal growth factor in mouse abdominal effusion

Epidermal growth factor (mEGF)-like immunoreactive material(s) was identified in mouse abdominal effusion (approximately 2.1 ng/mg protein) by our enzyme immunoassay (EIA) for mEGF. This material(s) and mEGF from the submaxillary glands of male mice were virtually equivalent with respect to the molecular weight and the antigenicity. Also, on isoelectric focusing analysis, the mEGF-like material(s) identified in abdominal effusion gave a major peak at pH 4.2 and a minor one at pH 4.5. These results demonstrate that the mEGF-like material(s) found in abdominal effusion is a molecule identical to mouse submaxillary gland EGF. Further we found that sialoadenectomy did not cause a marked decrease in the level of mEGF in abdominal effusion, suggesting that the source of mEGF found in abdominal effusion is other than the submaxillary glands.     

Kinematic analysis of bipedal locomotion of a Japanese macaque that lost its forearms due to congenital malformation

Spontaneously acquired bipedal locomotion of an untrained Japanese monkey (Macaca fuscata) is measured and compared with the elaborated bipedal locomotion of highly trained monkeys to assess the natural ability of a quadrupedal primate to walk bipedally. The subject acquired bipedalism by himself because of the loss of his forearms and hands due to congenital malformation. Two other subjects are performing monkeys that have been extensively trained for bipedal posture and locomotion. We videotaped their bipedal locomotion with two cameras in a lateral view and calculated joint angles (hip, knee, and ankle) and inertial angle of the trunk from the digitized joint positions. The results show that all joints are relatively more flexed in the untrained monkey. Moreover, it is noted that the ankle is less plantar flexed and the knee is more flexed in mid-to-late stance phase in the untrained monkey, suggesting that the trunk is not lifted up to store potential energy. In the trained monkeys, the joints are extended to bring the trunk as high as possible in the stance phase, and then stored potential energy is exchanged for kinetic energy to move forward. The efficient inverted pendulum mechanism seems to be absent in the untrained monkeys locomotion, implying that acquisition of such efficient bipedal locomotion is not a spontaneous ability for a Japanese monkey. Rather, it is probably a special skill that can only be acquired through artificial training for an inherently quadrupedal primate.This revised version was published online in April 2005 with corrections to the cover date of the issue.     

A vector system for genomic FLAG epitope-tagging in Schizosaccharomyces pombe

The fission yeast Schizosaccharomyces pombe is a popular model organism to study various cellular processes, although research tools available for S. pombe are relatively inadequate. To facilitate genetic and biochemical investigation in S. pombe, we report here a system of vectors for genomic FLAG epitope-tagging. These vectors enable us to amplify gene-targeting fragments for integration into specific loci of the S. pombe genome. All vectors in this report were designed to express FLAG epitope-tagged proteins from their endogenous genomic loci. Vectors for N-terminal FLAG epitope-tagging allow us to control protein expression levels using the wild-type nmt1 promoter, its weaker derivatives, and the urg1 promoter. These vectors are available with various antibiotic markers including kanMX6, hphMX6, natMX6 and bleMX6, and the his3(+) marker. Vectors for C-terminal FLAG epitope-tagging were designed to express FLAG-fusion proteins under the control of their native promoters at their own genomic loci, allowing us to characterize protein functions under physiological conditions. These vectors are available with kanMX6, hphMX6, nat-MX6 and bleMX6 markers. The series of vectors described in this report should prove useful for protein studies in fission yeast.     

Estimating the Functional Axis of the Primate Foot Using the Distribution of Plantar Muscles

Morton (American Journal of Physical Anthropology 5, 305–336, 1922) used the longest metatarsal, which he assumed functions as a lever during locomotion, to define the functional axis of the primate foot. In humans and apes, the functional foot axis lies on the second digit, whereas that of nonhominoid anthropoids is mostly on the third digit, suggesting that a medial shift of the functional axis occurred during primate foot evolution. Myological observations support this idea; the dorsal interossei of the human foot are arranged around the second digit, whereas those of nonhominoid anthropoids are around the third digit. However, it is still unclear when, why, and how such a change in foot musculature occurred. In addition, there is inconsistency among the limited number of studies that have examined foot musculature in apes. We examined modifications in the interosseous muscles of the chimpanzee, gibbon, spider monkey, and Japanese macaque in terms of the shift in the functional foot axis. We found that the dorsal interossei are arranged around the third digit; this is true even in the chimpanzee, whose functional axis based on metatarsal length lies on the second digit. This suggests that the change in the arrangement of the interosseous muscles phylogenetically lagged behind the shift of the osteological axis. Our results also indicate that the dorsal interossei are composite muscles consisting of the deep short flexors and the intermetatarsal abductors. We postulate that changes in the contributions of these 2 components to the formation of dorsal interossei likely occurred in the hominin lineage, resulting in the medial shift of the myological axis. The medial shift of the functional foot axis may have started with the elongation of the second metatarsal in the hominoid ancestors’ lineage, and was completed on the rearrangement of the interosseous muscles.     

Palmar and Plantar Pressure While Walking on a Horizontal Ladder and Single Pole in <Emphasis Type=Italic>Macaca fuscata</Emphasis>

To investigate biomechanical function in the hand and foot during quadrupedal locomotion in nonhuman primates, physical anthropologists and primatologists measure the pressure under them. We collected hand and foot pressure data while a Japanese macaque (Macaca fuscata), a semiterrestrial anthropoid, walked on 2 different simulated arboreal substrates, a horizontal ladder and a single pole, to explore differences in hand and foot use between the 2 substrates. The ladder rungs were perpendicular to the craniocaudal axis of the subject, and the pole was parallel to the subject’s craniocaudal axis. We tested the hypothesis that the pole was a more challenging substrate for the macaque than the ladder. Focusing on a diagonal sequence, diagonal couplets gait, we calculated gait characteristics and computed mean peak-pressure images of the hand and foot for each substrate from individual peak images via translation registration. We found several substrate differences that supported the hypothesis. The Japanese macaque walked at significantly slower speeds when traveling on the pole than on the ladder. Slower travel speed on the pole suggests that the Japanese macaque needed a wider support base to maintain balance on this substrate. Mean peak-pressure images suggest that the ladder invoked a more stepping-like behavior, but the pole invoked a more grasping-like behavior, especially of the foot. We show that the hand and foot use of the Japanese macaque would be adaptable to biomechanical challenges posed by different substrates.     

Roles of ChlR1 DNA helicase in replication recovery from DNA damage

The ChlR1 DNA helicase is mutated in Warsaw breakage syndrome characterized by developmental anomalies, chromosomal breakage, and sister chromatid cohesion defects. However, the mechanism by which ChlR1 preserves genomic integrity is largely unknown. Here, we describe the roles of ChlR1 in DNA replication recovery. We show that ChlR1 depletion renders human cells highly sensitive to cisplatin; an interstrand-crosslinking agent that causes stalled replication forks. ChlR1 depletion also causes accumulation of DNA damage in response to cisplatin, leading to a significant delay in resolution of DNA damage. We also report that ChlR1-depleted cells display defects in the repair of double-strand breaks induced by the I-PpoI endonuclease and bleomycin. Furthermore, we demonstrate that ChlR1-depeleted cells show significant delays in replication recovery after cisplatin treatment. Taken together, our results indicate that ChlR1 plays an important role in efficient DNA repair during DNA replication, which may facilitate efficient establishment of sister chromatid cohesion.