A physical domain of herpes simplex virus ICP8 is expressed and active in Escherichia coli.

In this report, we describe a series of procedures to assay the function of fusion genes in Escherichia coli and the specific application to the carboxy-terminal third of the herpes simplex virus type 1 (HSV-1) DNA-binding protein ICP8. E. coli cells containing the cloned HSV-1 BamHI G fragment with the HSV-1 BamHI-G-V site, map unit 0.388, nearest the tet promoter in pBR322 synthesized an active product containing a portion of ICP8. The new product induced phenotypic alterations in recipient hosts that were measurable and stable yet limited to the stability of the plasmid. The corresponding cloned DNA from the characterized HSV-1 DNA-binding protein mutant tsHA1 exhibited a predictable temperature-sensitive phenotype. Screening procedures based on the loss of induction of the parental plasmid-induced phenotype in E. coli cells allowed us to select additional mutations. One of these, which conferred a phenotype different from that of tsHA1, was transferred to the viral genome by marker transfer techniques. We suggest that any mutant could be isolated in any sequence, provided that the wild-type coding sequences induce alterations in E. coli cells. The observed alterations should have relevance in determining the mode of action of the protein in its normal environment.     

B cells in patients with X-linked agammaglobulinemia

X-linked agammaglobulinemia (XLA) has been described as a disorder in which pre-B cells fail to differentiate into B cells. However, a small number of B cells have been seen occasionally in patients with this disorder. Because the phenotype of these cells might be helpful in defining the site of the defect in XLA, immunofluorescent staining techniques were used to characterize the B cells that can be found in patients with XLA. Surface IgM-positive B cells could be detected in the peripheral circulation of all seven patients studied. These B cells constituted a very small percentage of the total lymphocytes (0.01 to 0.3% compared with 3.2 to 13.7% in controls) and differed in phenotype from control B cells. They were much more brightly stained for surface IgM (p less than 0.001) and less brightly stained for Ia (p less than 0.01). This phenotype is similar to that described for immature B cells in the mouse. Over 80% of the patients' B cells expressed surface IgD, and all expressed the B cell marker B1, but only 35% expressed the B cell marker B2. This B cell marker, which is the C3d receptor and the Epstein-Barr virus receptor, is expressed later in ontogeny than B1 and can be detected on over 80% of control B cells. All B cells expressed either kappa or lambda light chain. These findings indicate that the defect in differentiation of pre-B cells into B cells is not absolute in patients with XLA. The immature phenotype of the B cells additionally suggests that there may be a block in the maturation of B cells at more than one stage of differentiation in this disorder.     

Burn Injury Reduces Neutrophil Directional Migration Speed in Microfluidic Devices

Thermal injury triggers a fulminant inflammatory cascade that heralds shock, end-organ failure, and ultimately sepsis and death. Emerging evidence points to a critical role for the innate immune system, and several studies had documented concurrent impairment in neutrophil chemotaxis with these post-burn inflammatory changes. While a few studies suggest that a link between neutrophil motility and patient mortality might exist, so far, cumbersome assays have prohibited exploration of the prognostic and diagnostic significance of chemotaxis after burn injury. To address this need, we developed a microfluidic device that is simple to operate and allows for precise and robust measurements of chemotaxis speed and persistence characteristics at single-cell resolution. Using this assay, we established a reference set of migration speed values for neutrophils from healthy subjects. Comparisons with samples from burn patients revealed impaired directional migration speed starting as early as 24 hours after burn injury, reaching a minimum at 72–120 hours, correlated to the size of the burn injury and potentially serving as an early indicator for concurrent infections. Further characterization of neutrophil chemotaxis using this new assay may have important diagnostic implications not only for burn patients but also for patients afflicted by other diseases that compromise neutrophil functions.     

Critical manifolds,travelling waves,and an example from population genetics

A generalized Morse index theory is used to study travelling waves in a natural selection-migration model for a diploid organism when the selective strength is weak.     

The Invisibility of Disability: Using Dance to Shake from Bioethics the Idea of ‘Broken Bodies’

Complex social and ethical problems are often most effectively solved by engaging them at the messy and uncomfortable intersections of disciplines and practices, a notion that grounds the InVisible Difference project, which seeks to extend thinking and alter practice around the making, status, ownership, and value of work by contemporary dance choreographers by examining choreographic work through the lenses of law, bioethics, dance scholarship, and the practice of dance by differently‐abled dancers. This article offers a critical thesis on how bioethics has come to occupy a marginal and marginalizing role in questions about the differently‐abled body. In doing so, it has rendered the disabled community largely invisible to and in bioethics. It then defends the claim that bioethics – as a social undertaking pursued collaboratively by individuals from different disciplines – must take much better notice of the body and the embodied individual if it is to better achieve its ends, which include constructing a moral and just society. Finally, this article considers how the arts, and specifically dance (and here dance by differently‐abled dancers), provides us with rich evidence about the body and our ability to respond positively to normally ‘othered’ bodies. It concludes that greater attention to empirical evidence like that being generated in InVisible Difference will help to expand the reach and significance of bioethics, and thereby its relevance to (and consciousness of) important questions about the status of bodies and bodily differences, which must be considered as central to its ambitions.     

Inhibition of sperm transport and fertilization in superovulating ewes

In Experiment 1, all ewes were treated with follicle stimulating hormone (FSH-P) to induce superovulation. Ewes came into natural estrus or were treated with prostaglandin F(2)alpha (PGF(2)alpha) or 6-methyl-17-acetoxyprogesterone (MAP) to regulate the time of estrus. The ewes were mated during estrus and necropsied 3 h after mating. Regulation of estrus with either compound reduced the number of sperm recovered from the cervix, uterus, and oviducts and increased the proportions of sperm recovered from the cervix and uterine body that were immotile, dead, or had disrupted membranes. In Experiment 2, all ewes were in natural estrus. They either ovulated naturally or were superovulated, and ewes in each group were necropsied at 3 or 23 h after mating. Superovulation reduced the number of sperm in oviducts, uterus, and anterior segments of the cervix at both time intervals and increased the proportions of sperm that were immotile, dead, or had disrupted membranes. In Experiment 3, of 3x2 design, ewes were in either natural estrus or estrus regulated with PGF(2)alpha or with MAP; they ovulated naturally or were superovulated. Ewes were necropsied 3 d after mating and ova were examined. Both regulation of estrus and superovulation reduced the proportion of ova that were fertilized and reduced the number of accessory sperm attached to fertilized ova.     

Primary neoplasms of the facial nerve

A series of 30 primary facial nerve tumors is reviewed. Most of them were benign (n = 26); there were four malignant tumors. Neoplasms originating within the temporal bone were found to have preoperative facial paralysis in 84 percent of cases; the extracranial tumors had a 35 percent incidence of preoperative facial paralysis. All tumors in this series were treated surgically--by means of a middle fossa or transmastoid approach for the intratemporal group of tumors; the extracranial tumors were removed by the technique of parotid tumor surgery with complete facial nerve dissection. All the patients with preoperative facial weakness required facial nerve transection. Facial paralysis was rehabilitated with nerve grafts, hypoglossal crossover, or muscle transfers. Because "normal" facial expression is still not attainable following repair of complete facial nerve transection, an early diagnosis, hopefully prior to total neurotmesis, is essential. All patients with unexplained facial weakness, especially that which is progressive and persistent, should have the entire course of the facial nerve investigated for the possibility of treatable etiology.