Internalization of near-infrared fluorescent dyes within isolated macrophage populations.

The development and application of microsensor technology has enhanced the ability of scientists to further understand various biological activities, such as changes in the intracellular environment after injury or toxic exposure. NIR microsensor technology may be useful in detecting the cellular injuries or adverse changes during the early onset period, allowing for the administration of therapies to initiate recovery. The development and use of Infrared (IR) and near infrared (NIR) dyes as biological micro-sensors due to their advanced spectral characteristics may be helpful. Three of the more useful NIR dye characteristics include the ability to minimize background interference by extraneous biological matrices, the ability to exhibit optimal molar absorptivity and quantum yields, and the ability to maintain normal cellular activity. Thus, the current study was designed to investigate the ability of selected NIR micro-sensor dyes to undergo cellular internalization, demonstrate intracellular NIR fluorescent signaling, and maintain normal cellular activity. The results demonstrate that the selected NIR micro-sensor dyes undergo cellular internalization. The presence of the dyes within the cells did not affect cell viability. In addition, these dyes demonstrate changes in absorbance and fluorescence after the immune cells were challenged with a stimulant. Moreover, critical cellular functions, such as tumor necrosis factor release and superoxide production were not compromised by the internalization of the fluorescent dyes. These data suggest that selected NIR micro-sensor dyes can undergo intracellular internalization within isolated macrophages without adversely affecting various parameters of normal cellular activity.

Laparoscopic evaluation and management of a child with ambiguous genitalia, ectopic spleen, and Meckel's diverticulum.

Laparoscopy may be helpful in the evaluation and management of the child with intersex. Laparoscopic excision of dysgenetic gonads may be necessary due to the increased risk of malignancy. Residual müllerian duct structures are removed to prevent symptomatic complications at puberty. We present the case of a laparoscopic evaluation and management of a 46XY child with ambiguous genitalia, as well as the laparoscopic management of two unrelated anomalies discovered at the time of surgery.

Neurokinin induced inositol phosphate production in guinea pig bladder.

PURPOSE: To examine second messenger pathways involved in neurokinin induced bladder contractions. MATERIALS AND METHODS: Neurokinin induced changes in inositol phosphate production and in adenylyl cyclase activity are measured in the guinea pig bladder. RESULTS: Substance P, substance P methyl ester, neurokinin A, and neurokinin B each increase [3H]-inositol phosphate production in the guinea pig bladder. Substance P (10(-6) M) increases [3H]-inositol trisphosphate levels within 30 sec. Substance P and neurokinin A have an additive effect on inositol phosphate production, however substance P (10(-5) M) or neurokinin A (10(-5) M) induced inositol phosphate production is less than that induced by carbachol (10(-5) M). Neurokinin B and to a lesser extent neurokinin A inhibit forskolin-activated adenylyl cyclase activity. CONCLUSIONS: These data are compatible with neurokinin-induced inositol phosphate production being coupled to increases in contractile force of the guinea pig urinary bladder, however more than one second messenger pathway may be involved.

Effect of carbachol and norepinephrine on phosphatidyl inositol hydrolysis and cyclic AMP levels in guinea pig urinary tract.

Muscarinic cholinergic and adrenergic agonist-induced changes in [3H]-phosphatidyl inositol (PI) hydrolysis and cyclic AMP (cAMP) levels were measured in guinea pig ureter, urethra and bladder dome. In the ureter, carbachol, norepinephrine and phenylephrine rapidly increased PI hydrolysis and basal cAMP levels, but did not decrease forskolin-stimulated cAMP levels. In the bladder dome, norepinephrine and phenylephrine produced a rapid but transitory increase in PI hydrolysis, but did not affect forskolin-stimulated cAMP levels. Carbachol produced a rapid and sustained increase in PI hydrolysis and also, at high concentrations, decreased forskolin-stimulated cAMP levels. In the urethra, norepinephrine and carbachol rapidly decreased forskolin-stimulated cAMP levels and later increased PI hydrolysis. Our data suggest that the predominant second messenger system in the ureter, dome, or urethra is more dependent on the tissue than on the agonist. These tissue-specific, agonist-induced rapid changes in second messenger levels may help coordinate the contraction-relaxation phenomena necessary for urinary tract function.