Ser-Gln sites of SOG1 are rapidly hyperphosphorylated in response to DNA double-strand breaks.

The DNA damage response system (DDR) is crucial in addressing DNA double-strand breaks (DSBs), which pose a severe threat to genomic integrity. The SOG1 transcription factor is a master regulator of the Arabidopsis thaliana DDR. We previously showed that hyperphosphorylation of five Ser-Gln sites of SOG1 is the molecular switch to activate the DDR. In this study, we determined that SOG1 is hyperphosphorylated within 20 minutes following DSB-inducing treatment, followed by activation of several SOG1 target genes. Using SOG1 phosphorylation mutants, we demonstrated that although the hyperphosphorylation sites remain unchanged over time, the amount of hyperphosphorylation gradually increases. These observations suggest that rapid SOG1 hyperphosphorylation is limited by the amount of active kinases. ABBREVIATIONS: SOG1, suppressor of gamma response; ATM, Ataxia telangiectasia mutated; ATR, ATM and Rad3-related.

Contrasting size evolution in marine and freshwater diatoms.

Diatoms are key players in the global carbon cycle and most aquatic ecosystems. Their cell sizes impact carbon sequestration and energy transfer to higher trophic levels. We report fundamental differences in size distributions of marine and freshwater diatoms, with marine diatoms significantly larger than freshwater species. An evolutionary game theoretical model with empirical allometries of growth and nutrient uptake shows that these differences can be explained by nitrogen versus phosphorus limitation, nutrient fluctuations and mixed layer depth differences. Constant and pulsed phosphorus supply select for small sizes, as does constant nitrogen supply. In contrast, intermediate frequency nitrogen pulses common in the ocean select for large sizes or the evolutionarily stable coexistence of large and small sizes. Size-dependent sinking interacts with mixed layer depth (MLD) to further modulate optimal sizes, with smaller sizes selected for by strong sinking and shallow MLD. In freshwaters, widespread phosphorus limitation, together with strong sinking and shallow MLD produce size distributions with smaller range, means and upper values, compared with the ocean. Shifting patterns of nutrient limitation and mixing may alter diatom size distributions, affecting global carbon cycle and the structure and functioning of aquatic ecosystems.

Directionality of DNA replication fork movement strongly affects the generation of spontaneous mutations in Escherichia coli.

Using a pair of plasmids carrying the rpsL target sequence in different orientations to the replication origin, we analyzed a large number of forward mutations generated in wild-type and mismatch-repair deficient (MMR(-)) Escherichia coli cells to assess the effects of directionality of replication-fork movement on spontaneous mutagenesis and the generation of replication error. All classes of the mutations found in wild-type cells but not MMR(-) cells were strongly affected by the directionality of replication fork movement. It also appeared that the directionality of replication-fork movement governs the directionality of sequence substitution mutagenesis, which occurred in wild-type cells at a frequency comparable to base substitutions and single-base frameshift mutations. A very strong orientation-dependent hot-spot site for single-base frameshift mutations was discovered and demonstrated to be caused by the same process involved in sequence substitution mutagenesis. It is surprising that dnaE173, a potent mutator mutation specific for sequence substitution as well as single-base frameshift, did not enhance the frequency of the hot-spot frameshift mutation. Furthermore, the frequency of the hot-spot frameshift mutation was unchanged in the MMR(-) strain, whereas the mutHLS-dependent mismatch repair system efficiently suppressed the generation of single-base frameshift mutations. These results suggested that the hot-spot frameshift mutagenesis might be initiated at a particular location containing a DNA lesion, and thereby produce a premutagenic replication intermediate resistant to MMR. Significant numbers of spontaneous single-base frameshift mutations are probably caused by similar mechanisms.

Processing of shape defined by disparity in monkey inferior temporal cortex.

Neurons in the monkey inferior temporal cortex (IT) have been shown to respond to shapes defined by luminance, texture, or motion. In the present study, we determined whether IT neurons respond to shapes defined solely by binocular disparity, and if so, whether signals of disparity and other visual cues to define shape converge on single IT neurons. We recorded extracellular activity from IT neurons while monkeys performed a fixation task. Among the neurons that responded to at least one of eight random-dot stereograms (RDSs) containing different disparity-defined shapes, 21% varied their responses to different RDSs. Responses of most of the neurons were positively correlated between two sets of RDSs, which consisted of different dot patterns but defined the same set of eight shapes, whereas responses to RDSs and their monocular images were not correlated. This indicates that the response modulation for the eight RDSs reflects selectivity for shapes (or their component contours) defined by disparity, although responses were also affected by dot patterns per se. Among the neurons that showed selectivity for shapes defined by luminance or disparity, 44% were activated by both cues. Responses of these neurons to luminance-defined shapes and those to disparity-defined shapes were often positively correlated to each other. Furthermore the stimulus rank, which was determined by the magnitude of responses to shapes, generally matched between these cues. The same held true between disparity and texture cues. The results suggest that the signals of disparity, luminance, and texture cues to define the shapes converge on a population of single IT neurons to produce the selectivity for shapes.

Cervico-mediastinal bronchogenic cyst occurring in the prenatal period: report of a case.

We experienced a case of cervico-mediastinal bronchogenic cyst in which a cervical cystic mass was detected by prenatal ultrasonography. On prenatal ultrasound, a unilocular, well-defined and hypoechoic mass was detected in the fetal neck. The baby was born by a normal vaginal delivery at 40 weeks of gestation, and had no respiratory distress. Radiological investigations demonstrated a cyst in the cervico-mediastinal region, which displaced the trachea to the left. At the age of 32 days, an elective resection was easily performed through a right inferior collar incision after first aspirating the contents of the cyst. Prenatal sonography showing abnormal findings is effective for identifying cysts in the perinatal period and allows for the timely resection of such cysts before respiratory distress occurs.

Disparity selectivity of neurons in monkey inferior temporal cortex.

The inferior temporal cortex (IT) of the monkey, a final stage in the ventral visual pathway, has been known to process information on two-dimensional (2-D) shape, color, and texture. On the other hand, the dorsal visual pathway leading to the posterior parietal cortex has been known to process information on location in space. Likewise, neurons selective for binocular disparity, which convey information on depth, have been found mainly in areas along the dorsal visual pathway. Here, we report that many neurons in the IT are also selective for binocular disparity. We recorded extracellular activity from IT neurons and found that more than half of the neurons changed their response depending on the disparity added. The change was not attributed to monocular responses or eye movements. Most neurons selective for disparity were "near" or "far" cells; they preferred either crossed or uncrossed disparity, and only a small population was tuned to zero disparity. Disparity-selective neurons were also selective for shape. Most preferred the same type of disparity irrespective of the shape presented. Disparity preference was also invariant for the fronto-parallel translation of the stimuli in most of the neurons. Finally, nearby neurons exhibited similar disparity selectivity, suggesting the existence of a functional module for processing of binocular disparity in the IT. From the above and our recent findings, we suggest that the IT integrates shape and binocular disparity information, and plays an important role in the reconstruction of three-dimensional (3-D) surfaces.

The neural basis of perceptual and conceptual word priming--a PET study.

Positron emission tomography scans were obtained in 13 normal subjects during perceptual and conceptual word priming tasks with the aim to investigate the neural system specific to the two priming conditions. In the prescan phase, subjects were primed perceptually or conceptually with two separate procedures, while in the scan phase, they performed the same stem completion task. Therefore we could compare the results of the two priming tasks in a direct manner. A fixation control task and a baseline task (completion of stems that did not correspond to previously seen words) were also given. A specific blood flow decrease was found in the left inferior temporal cortex in the perceptual word priming condition and in the left superior temporal / inferior parietal cortex in the conceptual word priming condition. Each blood flow change may reflect transient changes in the cortical areas subserving the processing of the perceptual and conceptual components of word priming.

DNA replication errors produced by the replicative apparatus of Escherichia coli.

It has been hard to detect forward mutations generated during DNA synthesis in vitro by replicative DNA polymerases, because of their extremely high fidelity and a high background level of pre-existing mutations in the single-stranded template DNA used. Using the oriC plasmid DNA replication in vitro system and the rpsL forward mutation assay, we examined the fidelity of DNA replication catalyzed by the replicative apparatus of Escherichia coli. Upon DNA synthesis by the fully reconstituted system, the frequency of rpsL-mutations in the product DNA was increased to 1.9x10(-4), 50-fold higher than the background level of the template DNA. Among the mutations generated in vitro, single-base frameshifts predominated and occurred with a pattern similar to those induced in mismatch-repair deficient E. coli cells, indicating that the major replication error was slippage at runs of the same nucleotide. Large deletions and other structural alterations of DNA appeared to be induced also during the action of the replicative apparatus.

ADAM family proteins in the immune system.

CD156 is a member of a family proteins characterized by a disintegrin and a metalloprotease domain (ADAM). These molecules are phylogenically conserved but have individual roles in a variety of cells. Here, Shunsuke Yamamoto and colleagues discuss data suggesting that ADAM family proteins have important roles in the immune system.

Which method of flicker perimetry is most effective for detection of glaucomatous visual field loss?

PURPOSE: The authors compared the efficacy of two different forms of flicker perimetry: temporal modulation perimetry (TMP), which measures contrast thresholds for a fixed temporal frequency, and critical flicker frequency (CFF), which measures the highest frequency for which flicker is detected at a fixed contrast. METHODS: The authors compared 16 patients with early to moderate glaucomatous visual field loss with 16 age-matched normal controls. Flicker stimuli consisted of 2 degrees diameter targets of 2 seconds in duration, presented in 44 locations throughout the central 30 degrees visual field. Flicker was presented within a cosine envelope to avoid temporal transients. For TMP, contrast sensitivity thresholds were measured for 8-Hz sinusoidal flicker; CFF thresholds were measured for a stimulus of 100% contrast. RESULTS: The results indicate that TMP and CFF produced similar test-retest reliability in normals. CFF had slightly better reliability in glaucoma patients. Receiver operating characteristic analysis revealed that TMP could provide better separation of normals and glaucoma patients than did CFF. Similar findings were obtained when the thresholds for both procedures were converted to Z scores. CONCLUSIONS: Both methods of flicker perimetry testing provide acceptable test-retest reliability, and both can distinguish normal subjects from glaucoma patients. However, TMP is more effective in separating normal subjects from glaucoma patients than CFF, suggesting that TMP is the method of choice for detecting glaucomatous damage using flicker perimetry.

Structure of the murine CD156 gene, characterization of its promoter, and chromosomal location.

The murine cell surface antigen mCD156 is a glycoprotein that is expressed in monocytic cell lines and consists of a metalloprotease domain, a disintegrin domain, a cysteine-rich domain, and an epidermal growth factor-like domain in the extracellular region. The mCD156 gene is composed of 24 exons and 23 introns and spans approximately 14 kilobases. The first exon encodes most of the signal peptide sequence, and the transmembrane region is encoded by a single exon (19). In contrast, the other regions are composed of multiple exons. Of these, exons 7-12 and 12-15 encode a metalloprotease domain and a disintegrin domain, respectively. Sequence analysis of the 5'-flanking DNA revealed many potential regulatory motifs. Chloramphenicol acetyltransferase analysis demonstrated that nucleotides at positions -183, -334, and -623 contained cis-acting enhancing elements in a mouse monocytic cell line, aHINS-B3. Nucleotides at positions -183 and -390 contained elements responsible for lipopolysaccharide (LPS) inducibility, although several other 5'-flanking regions were also involved in LPS responsiveness. Regions -202, -507, and -659 play a role in interferon-gamma inducibility. Some of the potential regulatory motifs and other unknown cis elements may be involved in the constitutive expression, and LPS and interferon-gamma inducibilities. The mCD156 gene was mapped to chromosome 7, region F3-F4.

CD156 (human ADAM8): expression, primary amino acid sequence, and gene location.

The murine cell-surface antigen MS2 [ADAM 8; mouse CD156 (mCD156)] is a transmembrane glycoprotein that is highly expressed in monocytic lineages. mCD156 consists of a long exterior region containing domains strikingly similar to those of hemorrhagic snake venom proteins. cDNA for human CD156 was isolated from cDNA libraries from the human macrophage cell line THP-1 and from human granulocytes. The CD156 cDNA detected mRNA from human macrophage cell lines, granulocytes, monocytes, and B cell but not T cell lines. The nucleotide sequence of the CD156 cDNA showed 65.6% homology with that of mCD156, and its amino acid sequence had high homology with hemorrhagic snake venom proteins and other related mammalian proteins. The CD156 gene was mapped to human chromosome 10q26.3.

Disseminated Trichosporon infection. A case report and immunohistochemical study.

An autopsy case of disseminated Trichosporon beigelii infection in a patient with acute promyelocytic leukemia is presented. We diagnosed the T beigelii infection with immunoperoxidase method using our rabbit antiserum to T beigelii. The diagnosis of T beigelii infection is difficult, because of its close resemblance to Candida species in both clinical features and histopathologic findings. We could consistently identify T beigelii in tissue sections with the present immunohistologic method. We conclude that the immunoperoxidase method using antiserum to T beigelii is very useful to diagnose T beigelii infection.

Mapping of a gene determining tuberous sclerosis to human chromosome 11q14-11q23.

Tuberous sclerosis (TSC) is a dominantly inherited disorder characterized by hamartomas and hamartias in one or more organs, most often in skin, brain, and kidneys. Analysis of the basic genetic defect in tuberous sclerosis would be greatly expedited by definitive determination of the chromosomal location of the TSC gene or genes. We have carried out genetic linkage studies in 15 TSC families, using 34 polymorphic markers including protein markers and DNA markers. Pairwise lod scores were calculated using LIPED, and multipoint analyses were carried out using MENDEL. In the pairwise linkage analysis, using a penetrance value of 90%, a significant positive lod score was obtained with MCT128.1 (D11S144), 11q22-11q23, Zmax 3.26 at theta = 0.08. The tyrosinase probe TYR (11q14-11q22) gave a maximum lod score of 2.88 at theta = 0. In the multipoint analyses the most likely order is (TYR,TSC)-MCT128.1-HHH172. Homogeneity analysis was carried out using the USERM9 subprogram of MENDEL, which conducts the admixture test of C. Smith (1963, Ann. Hum. Genet. 27: 175-182). This test provided no evidence for genetic heterogeneity (that is, non-11-linked families) in this data set.

Growth rate of myelin figures of egg-yolk phosphatidylcholine.

The growth length of myelin figures was measured in detail using a video tape recording system of optical microscopy. In the beginning stage of growth, the growth process was not adapted to the diffusion-limited process of lipid molecules, which has been recently proposed. Another interpretation for the growth mechanism was proposed, where the growth results from swelling. The initial growth rate measured was in good agreement with the estimated value in consideration with the water flux in the first approximation of the lipid concentration gradient.

Emergency medical services and the pediatric patient: are the needs being met?

Emergency medical systems are being developed throughout the United States primarily to deal with myocardial infarction and trauma. These programs often fail to recognize the special needs of the critically ill child. Data collected in Los Angeles County from the LA County Trauma Surveys, Mobile Intensive Care Unit Rescue Reports, and Base Station Hospitals demonstrate that children represent approximately 10% of the paramedic calls. The calls are for medical problems as well as trauma. These data suggest that children have a higher death rate in the field than adults, and deaths occur more commonly in areas where there are no pediatric centers. Children are often secondarily transferred from emergency departments to other centers for definitive care. This study suggests that the needs of children in the prehospital setting are not being met.