A direct requirement of nuclear factor-kappa B for suppression of apoptosis in ventricular myocytes.

Nuclear factor-kappa B (NF-kappa B) is a ubiquitously expressed cellular factor regulated by the cytoplasmic factor inhibitor protein kappa B alpha (I kappa B alpha). Activation of NF-kappa B by cytokines, including tumor necrosis factor-alpha (TNF-alpha), requires the phosphorylation and degradation of I kappa B alpha. An anti-apoptotic role for NF-kappa B has recently been suggested. In the present study, we ascertained whether death-promoting signals and apoptosis mediated by TNF-alpha are suppressed by NF-kappa B in postnatal ventricular myocytes. Stimulation of myocytes with TNF-alpha resulted in a 12.1-fold increase (P < 0.01) in NF-kappa B-dependent gene transcription and DNA binding compared with controls. This was accompanied by a corresponding increase in the NF-kappa B target protein A20 as determined by Western blot analysis. Vital staining revealed that TNF-alpha was not cytotoxic to myocytes and did not provoke apoptosis. Adenovirus-mediated delivery of a nonphosphorylatable form of I kappa B alpha to inactivate NF-kappa B prevented TNF-alpha-stimulated NF-kappa B-dependent gene transcription and nuclear NF-kappa B DNA binding. Importantly, myocytes stimulated with TNF-alpha and defective for NF-kappa B activation resulted in a 2.2-fold increase (P < 0.001) in apoptosis. To our knowledge, the data provide the first indication that a functional NF-kappa B signaling pathway is crucial for suppressing death-promoting signals mediated by TNF-alpha in ventricular myocytes.

The effects of caffeine on near point plus acceptance.

Forty-two university students participated in a study of reading with +0.50 sphere or yokes prism base down for overconvergence and base up for underconvergence. Sixty-four percent of the group improved with a mean of +10.79 wpm. The level of coffee consumption of each student was recorded. A negative correlation existed between the intake of caffeine and reading improvement, r = -.81, p less than .001. Two students reduced their coffee consumption and transformed their loss in words per minute to a significant gain. A double-blind protocol was used in this study. A placebo lens (plano) controlled for subject and investigator bias. The relationship between caffeine and accommodation was investigated by means of a monocular test through a +4.D sphere. The location of the blur point was indicative of the state of tonicity of the ciliary system. Pre- and post-tests of the blur point for 14 subjects who were tested before and after consuming 150 mg of caffeine showed a significant change. The experimental group showed an increase of -0.52D while the control group showed a change of +0.36D. r = .781, p less than .01.

Stability of bovine adrenal medulla cells in culture.

The functional stability of primary cultures of adrenal medulla cells was investigated. Isolated cells were prepared by treatment of bovine adrenal glands with collagenase followed by purification on Percoll density gradients and were maintained in Dulbecco's medium containing 10% fetal calf serum. Within 12 h after plating on plastic culture dishes, the cells became firmly attached and exhibited good survival for periods of time up to 3 weeks, as indicated by their morphology using light and electron microscopy, by maintenance of their content of catecholamines, tyrosine hydroxylase, dopamine-beta-hydroxylase, and phenylethanolamine N-methyltransferase, and their ability to respond to secretagogues. During the first 10 days to 2 weeks in culture there was little or no change in any of these parameters. During the 3rd week there were progressive losses of catecholamine and enzyme activities and increased vacuolization of medullary cells. The cells synthesized protein and RNA with no apparent loss in activities over the period studied, but did not incorporate [3H]thymidine into PCA-precipitable material. The cells responded to secretagogues and secretory antagonists similarly to isolated perfused adrenal glands. The studies described here demonstrate that primary cultures of adrenal medulla cells provide an excellent experimental system for obtaining more detailed information on stimulus-secretion coupling and other functional aspects of the adrenal medulla.

Long-Term Activity Recording in small Aquatic Animals.

Figure 1 of the paper " Long-term activity in small aquatic animals" by A. A. Heusner and J. T. Enright (28 Oct. 1966, p. 532) is ambiguous to the extent that the two negative terminals might be interpreted as a common point in the circuit. In fact, these two points must be kept separate for the proper operation of the instrument.

Release of catecholamines and specific protein from adrenal glands.

Rabbit antiserums directed against a purified protein obtained from the catecholamine storage vesicles of the adrenal medulla of the cow were used to study the mechanism of secretion from the adrenal medulla. During secretion evoked by acetylcholine in isolated perfused adrenal glands, catecholamines and the specific intravesicular protein were released in relative amounts comparable to those present in the intact gland. Thus catecholamines are apparently released from the storage vesicles by a process resembling reverse pinocytosis.