Assessment of students' attitudes toward breastfeeding.

A self-administered survey questionnaire was distributed to 346 high school and 244 college students in Alabama to explore their perceptions of breastfeeding. Only 135 acknowledged having been breastfed. Embarrassment was perceived as a major barrier to breastfeeding; less than half thought breastfeeding should be done publicly. However, respondents had generally positive attitudes about breastfeeding. They intended to support breastfeeding of their own child; thought that breastfeeding was more healthful than bottle-feeding and more convenient; and that breastfeeding is not obscene nor does it make a woman less attractive. Over half received breastfeeding information from home, school, and television. Further, both high school and college students supported breastfeeding education in schools. These findings suggest that although fears of embarrassment is a major barrier to breastfeeding, the students showed overall positive attitudes about breastfeeding despite the region's low breastfeeding rate. Breastfeeding promotional programs should address the stigma of embarrassment associated with breastfeeding.

Discordant hepatic expression of the cell division control enzyme p34cdc2 kinase, proliferating cell nuclear antigen, p53 tumor suppressor protein, and p21Waf1 cyclin-dependent kinase inhibitory protein after WY14,643 ([4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid) dosing to rats.

The hepatocarcinogen and peroxisome proliferator WY14,643 ([4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid) was examined for its ability to induce changes in the intracellular protein expression of hepatic p34cdc2 kinase (CDK1), proliferating cell nuclear antigen (PCNA), p53 tumor suppressor protein, and p21Waf1 CDK inhibiting protein. Young adult male rats were administered 45 mg-kg/day WY14,643 intraperitoneally for 1, 2, 3, 4, or 5 days or fed diets containing 0% or 0.08% WY14,643 for 1, 2, 3, or 4 weeks. WY14,643 dosing increased concentrations of hepatic proteins of 34- and 37-kDa molecular mass, which were identified through immunoprecipitation as CDK1 and PCNA, respectively. Gel filtration of the hepatic S9 fractions determined by enzyme-linked immunosorbent assay confirmed the increased expression of CDK1 and PCNA immunoreactivity in livers from WY14,643-treated rats. Also, gel filtration revealed that the native CDK1 and PCNA in hepatic S9 from WY14,643-treated rats chromatographed as a major peak with an apparent molecular mass of 70 and 76 kDa, respectively. Immunoblotting of the 70-kDa fraction with anti-CDK1 revealed a single band of molecular mass of 34 kDa. Thus, the CDK1 in the major immunoreactive peak of WY14,643-treated rat liver S9 seems to exist as a heterodimer or homodimer. Immunohistochemistry of formalin-fixed liver demonstrated a cytosolic localization of immunoreactive CDK1 and nuclear localization of immunoreactive PCNA in proliferating cells of WY14,643-treated rat livers. WY14,643 increased hepatic CDK1 content by 1.9-6.3-fold through postdosing days 1-5. Hepatic PCNA content was increased 1.9-5-fold over the same period. In the 4-week feeding study, CDK1 and PCNA expression were increased at all weekly time points by an average of 15-50-fold, respectively. Furthermore, the dietary administration of 0.08% WY14,643 resulted in sustained, overexpression of hepatic p53 tumor suppressor protein from week 1 through week 4 and of p21Waf1 CDK inhibitory protein from week 3 to week 4.

Discordant expression of the cyclin-dependent kinases and cyclins in rat liver following acute administration of the hepatocarcinogen [4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio] acetic acid (WY14,643).

Cellular proliferation is an essential aspect of chemical carcinogenesis. At the core of cell cycle regulation is a family of serine/threonine protein kinases termed cyclin-dependent kinases (cdk). Cdk activity, which directs progression through the cell cycle, is dependent upon cdk binding to the appropriate, phase-specific cyclin proteins. Alterations in hepatic cdk1, cdk2, cdk4, cdk5, and cyclin protein expression were determined in response to acute dosing of the prototypic peroxisome proliferator and hepatocarcinogen [4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio] acetic acid (WY14,643). Intraperitoneal dosing of 45 mg WY14,643/kg daily for 4 days to young, male rats produced dramatic increases in hepatic protein expression of all cdk analyzed as well as cyclins B, D2, D3, and proliferating cell nuclear antigen (PCNA). The largest relative increases, 6.1-, 2.8-, 11-, 83-, and 7.9-fold, were seen with cdk1, cdk4, cyclin B, cyclin D3, and PCNA, respectively. Increases of only 1.8-, 2-, 1.6-, and 1.4-fold were noted, respectively, for cdk2, cdk5, cyclin D2, and cyclin E. Analysis of gel filtration fractions indicated that PCNA co-eluted with cdk1 from the WY14,643-treated rats as a 70-80 kDa molecular complex. In contrast, cdk4, cdk5 and D cyclins migrated as much larger complexes with an estimated MW of approximately 180-190 kDa.

Detection of cytochrome b5 from the house-fly, Musca domestica: comparison of immunological and spectrophotometric methods.

Spectrophotometric assay of microsomal cytochrome b5 in house-flies produces different results depending on whether sodium dithionite or NADH is used as the reducing agent and whether or not detergent is present. Microsomes assayed for cytochrome b5 with dithionite in the presence of detergent gave the highest values, followed by dithionite alone, NADH plus detergent, and then NADH alone. Isopropanol treatment of microsomes extracted cytochrome b5 free of spectrophotometrically interfering cytochrome P-450. Studies using immunoblotting and rocket immunoelectrophoresis with polyclonal antisera raised against the purified cytochrome b5 showed that isopropanol treatment quantitatively extracted cytochrome b5.

Anti-P450lpr antiserum inhibits specific monooxygenase activities in LPR house fly microsomes.

Monooxygenase activity in microsomes from the LPR strain of house fly (Musca domestica L.) was inhibited by anti-P450lpr, and antiserum specific for house fly cytochrome P450lpr. Anti-P450lpr did not inhibit house fly cytochrome P450 reductase or rat cytochrome P450 monooxygenase assays, consistent with specific inhibition of P450lpr. Anti-P450lpr inhibited the ability of cytochrome P450 reductase to reduce carbon monoxide treated LPR microsomal cytochrome P450, up to 49% of the total, showing that inhibition of cytochrome P450 reduction is the major mechanism of inhibition. Anti-P450lpr inhibited 98% of methoxyresorufin-O-demethylase activity and all the benzo(a)pyrene hydroxylase activity in LPR microsomes, but none of the pentoxyresorufin-O-dealkylase activity. The antiserum partially inhibited ethoxyresorufin-O-dealkylase and ethoxycoumarin-O-dealkylase activity. These results demonstrate that methoxyresourfin-O-demethylase activity and benzo(a)pyrene hydroxylase activity are characteristic substrates for P450lpr activity in the LPR strain of house fly.

Rapid isolation of a neurohormone from mosquito heads by high-performance liquid chromatography.

Methods were developed for the isolation of the egg development neurosecretory hormone, EDNH, from heads of the mosquito Aedes aegypti. This hormone stimulates ecdysone production by ovaries. Methods used for the successful isolation of insulin-like peptides from vertebrate tissues were modified to develop a four-step procedure involving extraction in acidified ethanol, precipitation by neutralization, followed by sequential separation on size-exclusion, ion-exchange and reversed-phase high-performance liquid chromatography columns.

Expression of cytochrome P-450lpr is developmentally regulated and limited to house fly.

Expression of house fly cytochrome P-450lpr was examined using immunoblotting in male and female adult LPR house flies, mixed sex adult house flies at 12 different ages, larvae, and pupae. P-450lpr was expressed in both male and female adult house flies. P-4501pr was clearly present in all adult stages examined, was barely detectable in pupae, and could not be detected in larvae. Thus, cytochrome P-450lpr is developmentally regulated and present in both sexes of house fly. Expression of cytochrome P-450, immunologically homologous to house fly cytochrome P-4501pr, was examined in other species using immunoblot analysis. Eleven animal species were tested in the orders Diptera, Hymenoptera, Lepidoptera, Orthoptera, Acari, and Rodentia, using microsomes in some species from both induced and noninduced animals or insecticide-resistant and susceptible strains. P-450lpr appears to be restricted to house flies, as none of these species contained cytochrome P-450 that reacted with antiserum to cytochrome P-450lpr.

Egg maturation and ecdysiotropic activity in extracts of mosquito (Aedes aegypti) heads.

Blood-fed, decapitated female Aedes aegypti mosquitoes matured eggs when injected with an extract of heads, but non-blood-fed females did not. The response to head extract was optimal when the head was allowed to remain for 2 hr after feeding. A dose response to injected egg development neurosecretory hormone (EDNH) was observed in vivo that was similar to in vitro dose responses previously reported. Blood-fed decapitated females responded equally well to boiled or unboiled head extract. When blood-fed decapitated females were injected with head extract, ecdysteroid levels increased. Partial purification of head extract using high-pressure liquid chromatography yielded a fraction at 34% acetonitrile that showed egg maturation activity in vivo when injected into blood-fed decapitated females, and ecdysiotropic activity when incubated in vitro with ovaries. In addition, a fraction at 30% acetonitrile was found that showed activity in vivo but not in vitro and may be a precursor. Occasionally, the fraction at 37% acetonitrile showed activity in the in vitro assay but had little activity in vivo and may be a metabolite. These results suggest that the same hormone was being assayed in vivo and in vitro and is EDNH.