Production of Macrophage Migration Inhibition Factors by Virus-Infected Cell Cultures

Macrophage migration inhibitory (MIF-like) activity was demonstrated in the supernatant fluids from cultures of African green monkey kidney cells (BGM) infected with mumps virus or Newcastle disease virus. We could detect no such activity in noninfected cultures. The virus-induced activity reported here is not due to nonspecific cytotoxic material released by dead or dying cells, and it does not require cell replication for its production. Preliminary estimates of molecular weight by Sephadex G-100 chromatography revealed a broad band of activity associated with the 45,000 and 65,000 markers. These are significantly smaller than previously reported chemotactic substances from virus-infected cultures, and thus appear to represent different cell products. These MIF-like factors may be produced concomitantly with interferon. However, ultraviolet irradiation of appropriate duration abolishes the ability of viruses to induce substances with MIF-like activity while preserving the ability to induce interferon. This strongly suggests that interferon is not the agent responsible for the macrophage migration inhibition effect. The functional properties of these various cell products induced by virus infection suggest that they all may play a role in the response to virus infection in vivo.     

Detection of chromosomes and estimation of aneuploidy in human spermatozoa using fluorescence in-situ hybridization

The development and application of fluorescence in-situ hybridization (FISH) has opened the way for comprehensive studies on numerical chromosome abnormalities in human spermatozoa. FISH can be rapidly applied to large numbers of spermatozoa and thus overcomes the major limitation of karyotyping spermatozoa after penetration of zona-free hamster oocytes. The simultaneous hybridization of two or more chromosome-specific probes to spermatozoa and subsequent detection of the bound probes using different fluorescent detection systems enables two or more chromosomes to be localized simultaneously in the same spermatozoon and provides a technique for undertaking reasonable estimates of aneuploidy. The most commonly used probes are those which bind to the centromeric region of specific chromosomes. Most studies to date have concentrated on estimating aneuploidy in spermatozoa from normospermic men, although reports are beginning to appear on aneuploidy in spermatozoa from subfertile and infertile men. Multi- probe FISH studies have generally reported disomy (hyperhaploidy) estimates of 0.05-0.2% per chromosome. There is preliminary evidence that some chromosomes such as X, Y and 21 are predisposed towards higher rates of non-disjunction during spermatogenesis. There are also suggestions of inter-donor variability in aneuploidy frequencies for specific chromosomes, although this requires confirmation in larger studies. While FISH is clearly a powerful technique that has many applications in reproductive medicine, it must also be realized that it does have limitations and the technology itself is still evolving and has yet to be fully validated on spermatozoa.      

Interference between velocity-dependent and position-dependent force-fields indicates that tasks depending on different kinematic parameters compete for motor working memory

Humans demonstrate motor learning when exposed to changes in the dynamics of movement or changes in the visuomotor map. However, when two opposing dynamic transformations are learned in succession, the memory of the first is overwritten by learning of the second; the same is true for two opposing visuomotor rotations. This retrograde interference is not seen for all combinations of transformations, however. When a dynamic transformation is learned subsequent to a visuomotor rotation, the presence or absence of interference appears to depend crucially on the structure of the dynamic task: a force-field dependent on the position of the hand produces interference, whereas an inertial load applied lateral to the hand does not. To explain these results, it has been hypothesized that two transformations can be learned without interference if they depend on two different kinematic parameters of movement (such as position and velocity of the hand). Here we demonstrate, contrary to this hypothesis, interference between a dynamic transformation that depends on the position of the hand and one that depends on its velocity. However, the interference was found to be incomplete, supporting the view that the ability to retain motor memories for different tasks depends on the degree to which their representations conflict in working memory.     

Differential regulation of mast cell cytokines by both dexamethasone and the p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580

Activated mast cells generate multiple cytokines but it is not known if these can be differentially regulated by pharmacological agents. We report here that the glucocorticoid dexamethasone (DEX) preferentially inhibited Ag-induced expression of IL-4 and IL-6 mRNA relative to TNF-alpha mRNA in RBL-2H3 cells. Likewise, the drug more readily inhibited release of IL-4 than TNF-alpha protein. SB203580, an inhibitor of p38 mitogen-activated protein kinase (MAPK), enhanced Ag-induced TNF-alpha mRNA expression without affecting IL-4 or IL-6 mRNA. At the protein level, SB203580 exerted little effect on TNF-alpha release but inhibited IL-4 release; notably, the ratio of TNF-alpha : IL-4 increased markedly with the concentration of SB203580, confirming the differential regulation of these cytokines. PD98059, an inhibitor of MAPK kinase (MEK), a component of the p44/42 MAPK pathway, partially inhibited Ag-induced expression of mRNA for all three cytokines while cyclosporin A inhibited Ag-induced IL-4 and IL-6 mRNA more readily than TNF-alpha mRNA. Ag activation of the cells led to phosphorylation of p38 and p44/42 MAPK but this was not influenced by DEX. In conclusion, mast cell cytokines can be differentially regulated pre- and post-translationally by DEX and SB203580 but there does not appear to be a direct mechanistic link between the actions of these two drugs.     

Control of grasp stability in humans under different frictional conditions during multidigit manipulation

Control of grasp stability under different frictional conditions has primarily been studied in manipulatory tasks involving two digits only. Recently we found that many of the principles for control of forces originally demonstrated for two-digit grasping also apply to various three-digit grasps. Here we examine the control of grasp stability in a multidigit task in which subjects used the tips of the thumb, index, and middle finger to lift an object. The grasp resembled those used when lifting a cylindrical object from above. The digits either all contacted the same surface material or one of the digits contacted a surface material that was more, or less, slippery than that contacted by the other two digits. The three-dimensional forces and torques applied by each digit and the contact positions were measured along with the position and orientation of the object. The distribution of forces among the digits strongly reflected constraints imposed by the geometric relationship between the object's center of mass and the contact surfaces. On top of this distribution, we observed changes in force coordination related to changes in the combination of surface materials. When all digits contacted the same surface material, the ratio between the normal force and tangential load (F(n):L ratio) was similar across digits and scaled to provide an adequate safety margin against slip. With different contact surfaces subjects adapted the F(n):L ratios at the individual digits to the local friction with only small influences by the friction at the other two digits. They accomplished this by scaling the normal forces similarly at all digits and changing the distribution of load among the digits. The surface combination did not, however, influence digit position, tangential torque, or object tilting systematically. The change in load distribution, rather, resulted from interplay between these factors, and the nature of this interplay varied between trials. That is, subjects achieved grasp stability with various combinations of fingertip actions and appeared to exploit the many degrees of freedom offered by the multidigit grasp. The results extend previous findings based on two-digit tasks to multidigit tasks by showing that subjects adjust fingertip forces at each digit to the local friction. Moreover, our findings suggest that subjects adapted the load distribution to the current frictional condition by regulating the normal forces to allow slips to occur early in the lift task, prior to object lift-off.     

In ovarian neoplasms, BRAF, but not KRAS, mutations are restricted to low-grade serous tumours

Genes of the RAF family, which mediate cellular responses to growth signals, encode kinases that are regulated by RAS and participate in the RAS/RAF/MEK/ERK/MAP-kinase pathway. Activating mutations in BRAF have recently been identified in melanomas, colorectal cancers, and thyroid and ovarian tumours. In the present study, an extensive characterization of BRAF and KRAS mutations has been performed in 264 epithelial and non-epithelial ovarian neoplasms. The epithelial tumours ranged from adenomas and borderline neoplasms to invasive carcinomas including serous, mucinous, clear cell, and endometrioid lesions. It is shown that BRAF mutations in ovarian tumours occur exclusively in low-grade serous neoplasms (33 of 91, 36%); these included serous borderline tumours (typical and micropapillary variants), an invasive micropapillary carcinoma and a psammocarcinoma. KRAS mutations were identified in 26 of 91 (29.5%) low-grade serous tumours, 7 of 49 (12%) high-grade serous carcinomas, 2 of 6 mucinous adenomas, 22 of 28 mucinous borderline tumours, and 10 of 18 mucinous carcinomas. Of note, two serous borderline tumours were found to harbour both BRAF and KRAS mutations. The finding that at least 60% of serous borderline tumours harbour mutations in two members of the ERK-MAP-kinase pathway (BRAF 36%, KRAS 30%) compared with 12% of high-grade serous carcinomas (BRAF 0%, KRAS 12%) indicates that the majority of serous borderline tumours do not progress to serous carcinomas. Furthermore, no BRAF mutations were detected in the other 173 ovarian tumours in this study.