Human B1 Cell Frequency: Isolation and Analysis of Human B1 Cells

Controversy over the frequency of human B1 cells in normal individuals has arisen as different labs have begun to employ non-uniform techniques to study this population. The phenotypic profile and relative paucity of circulating human B1 cells place constraints on methodology to identify and isolate this population. Multiple steps must be optimized to insure accurate enumeration and optimal purification. In the course of working with human B1 cells we have developed a successful strategy that provides consistent analysis of B1 cells for frequency determination and efficient isolation of B1 cells for functional studies. Here we discuss issues attendant to identifying human B1 cells and outline a carefully optimized approach that leads to uniform and reproducible data

Systems biology analysis of drivers underlying hallmarks of cancer cell metabolism.

Malignant transformation is often accompanied by significant metabolic changes. To identify drivers underlying these changes, we calculated metabolic flux states for the NCI60 cell line collection and correlated the variance between metabolic states of these lines with their other properties. The analysis revealed a remarkably consistent structure underlying high flux metabolism. The three primary uptake pathways, glucose, glutamine and serine, are each characterized by three features: (1) metabolite uptake sufficient for the stoichiometric requirement to sustain observed growth, (2) overflow metabolism, which scales with excess nutrient uptake over the basal growth requirement, and (3) redox production, which also scales with nutrient uptake but greatly exceeds the requirement for growth. We discovered that resistance to chemotherapeutic drugs in these lines broadly correlates with the amount of glucose uptake. These results support an interpretation of the Warburg effect and glutamine addiction as features of a growth state that provides resistance to metabolic stress through excess redox and energy production. Furthermore, overflow metabolism observed may indicate that mitochondrial catabolic capacity is a key constraint setting an upper limit on the rate of cofactor production possible. These results provide a greater context within which the metabolic alterations in cancer can be understood

Hurricane Florence Flooding in Georgetown County: A Qualitative Explanation of the Interactions of Estuary and Tidal River

This paper examines data from 18 USGS gauges in the lower Pee Dee Basin in an effort to explain the behavior of the flooding following Hurricane Florence (2018) in Georgetown County, South Carolina. Despite record or near-record flooding in all the tributaries to the Winyah Bay estuary, water levels near the city of Georgetown were well below predicted heights. Floodplain storage in the lower Great Pee Dee, Lynches, and Little Pee Dee River valleys stored over 1.2 million acre-feet of floodwaters, delaying peak stage near Bucksport for five days and reducing peak flow into the Winyah Bay tidal river/estuary system by nearly 50%. An unknown amount of flow from the Winyah Bay tidal river/estuary system flowed through the Atlantic Intracoastal Water Way to Little River rather than through Winyah Bay. The resulting freshwater flow to Winyah Bay only moved the point of tidal stagnation (where upstream tidal flow balances downstream freshwater flow) to near Georgetown. Since the city of Georgetown was near the point of stagnation, water level there was driven by ocean tidal height rather than river flood stage. The lack of discharge data from the tidal rivers in Georgetown County prevents evaluation of the importance of each of these factors and will limit efforts to make quantitative predictions of future flooding in the county

Methodological Issues in Multistage Genome-Wide Association Studies

Because of the high cost of commercial genotyping chip technologies, many investigations have used a two-stage design for genome-wide association studies, using part of the sample for an initial discovery of ``promising'' SNPs at a less stringent significance level and the remainder in a joint analysis of just these SNPs using custom genotyping. Typical cost savings of about 50% are possible with this design to obtain comparable levels of overall type I error and power by using about half the sample for stage I and carrying about 0.1% of SNPs forward to the second stage, the optimal design depending primarily upon the ratio of costs per genotype for stages I and II. However, with the rapidly declining costs of the commercial panels, the generally low observed ORs of current studies, and many studies aiming to test multiple hypotheses and multiple endpoints, many investigators are abandoning the two-stage design in favor of simply genotyping all available subjects using a standard high-density panel. Concern is sometimes raised about the absence of a ``replication'' panel in this approach, as required by some high-profile journals, but it must be appreciated that the two-stage design is not a discovery/replication design but simply a more efficient design for discovery using a joint analysis of the data from both stages. Once a subset of highly-significant associations has been discovered, a truly independent ``exact replication'' study is needed in a similar population of the same promising SNPs using similar methods.Comment: Published in at http://dx.doi.org/10.1214/09-STS288 the Statistical Science (http://www.imstat.org/sts/) by the Institute of Mathematical Statistics (http://www.imstat.org

Suicide: A Constitutional Right?

Recent advocacy of a right to suicide raises the question of whether statutes which penalize the assistance of suicide and the widespread practice of intervention to stop suicide attempts are unconstitutional. This article considers the claim that the right of privacy or substantive due process encompasses autonomy to do whatever affects no one else, and concludes that the Supreme Court has recognized only those forms of autonomy that it has deemed to be rooted in the traditions and history of our society. After outlining the attitudes toward suicide at various stages in Western Civilization, focusing on Anglo- American history and particularly on the legal tradition, the authors conclude that the predominant approach has been to discourage suicide and to penalize its assistance, while, at least in American history, to treat rather than condemn the suicide attempter. In their view, suicide has not historically been treated as a fundamental right. The article then relates psychological and sociological evidence that those who attempt suicide are normally ambivalent, usually do so for reasons other than a settled desire to die, and are predominantly the victims of mental disorder. Finally, the authors argue that societal sanctions for suicide would lead to manipulation by others and social pressure that would induce many unstable individuals, who would otherwise be helped, to commit suicide. For these reasons, the authors regard recognition of a right to suicide as unjustified and undesirable

Nucleotide sequence of the luxA gene of Vibrio harveyi and the complete amino acid sequence of the alpha subunit of bacterial luciferase

The nucleotide sequence of the 1.85-kilobase EcoRI fragment from Vibrio harveyi that was cloned using a mixed-sequence synthetic oligonucleotide probe (Cohn, D. H., Ogden, R. C., Abelson, J. N., Baldwin, T. O., Nealson, K. H., Simon, M. I., and Mileham, A. J. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 120-123) has been determined. The alpha subunit-coding region (luxA) was found to begin at base number 707 and end at base number 1771. The alpha subunit has a calculated molecular weight of 40,108 and comprises a total of 355 amino acid residues. There are 34 base pairs separating the start of the alpha subunit structural gene and a 669-base open reading frame extending from the proximal EcoRI site. At the 3' end of the luxA coding region there are 26 bases between the end of the structural gene and the start of the luxB structural gene. Approximately two-thirds of the alpha subunit was sequenced by protein chemical techniques. The amino acid sequence implied by the DNA sequence, with few exceptions, confirmed the chemically determined sequence. Regions of the alpha subunit thought to comprise the active center were found to reside in two discrete and relatively basic regions, one from around residues 100-115 and the second from around residues 280-295