Chronic combined hyperandrogenemia and western-style diet in young female rhesus macaques causes greater metabolic impairments compared to either treatment alone.

STUDY QUESTION: Does developmental exposure to the combination of hyperandrogenemia and western-style diet (WSD) worsen adult metabolic function compared to either treatment alone? SUMMARY ANSWER: Young female rhesus macaques treated for 3 years, beginning at menarche, with combined testosterone (T) and WSD have increased weight gain and insulin resistance compared to controls and animals treated with either T or WSD alone. WHAT IS KNOWN ALREADY: Hyperandrogenemia is a well-established component of polycystic ovary syndrome (PCOS) and can be observed in peripubertal girls, indicating a potential pubertal onset of the disease. Obesity is often associated with hyperandrogenemia in peripubertal girls, and overweight girls appear to be at higher risk for the development of PCOS later in life. STUDY DESIGN, SIZE, DURATION: Juvenile (2.5- year old) female rhesus macaques were divided into four groups (n = 10/group): control animals receiving cholesterol implants and a control diet with 15% of calories derived from fat (C), animals receiving T implants (mean serum levels: 1.35 ± 0.01 ng/ml) and a control diet (T), animals receiving a cholesterol implant and a WSD with 36% of calories derived from fat (WSD) and animals receiving a T implant and a WSD (T + WSD). Animals were maintained on the treatments for 36 months and were 5.5 years old at study completion. PARTICIPANTS/MATERIALS, SETTING, METHODS: Metabolic testing consisted of body measurements including weight, dual-energy X-ray absorptiometry scans, activity monitoring, and glucose tolerance testing at zero months and at least once every 12 months for the remainder of the study. Indirect calorimetry and serum hormone assays were performed following 36 months of treatment. MAIN RESULTS AND THE ROLE OF CHANCE: Body weight and fat mass gain were significantly increased in T + WSD at 24 and 36 months of treatment compared to the other three groups. Log transformed fasting insulin and Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) were significantly increased in T + WSD animals at 3 years of treatment compared to all other groups. T-treatment caused a greater rate of decline in activity after 18 months, while food intake and metabolic rate were largely unaffected by treatments. LIMITATIONS REASONS FOR CAUTION: Variability was present in the metabolic parameters measured; however, this is similar to the heterogeneity observed in human populations. WIDER IMPLICATIONS OF THE FINDINGS: Chronic hyperandrogenemia beginning at puberty may exacerbate metabolic dysfunction in women consuming a WSD and account for the increased rates of obesity and insulin resistance observed in PCOS patients. Counseling of female patient populations with elevated androgens about the potential benefit of consuming a lower fat diet could improve long-term metabolic health outcomes. STUDY FUNDING/COMPETING INTEREST(S): Eunice Kennedy Shriver National Institute of Child Health & Human Development P50HD071836 and Oregon National Primate Center Grant P51 OD011092. The authors have no competing conflict of interests to disclose.

Induction of cytotoxic T cell responses in newborn mice by DNA immunization.

Cytotoxic T cells (CTL) play a critical role in controlling viral infections. Infection of neonatal NFSIN mice with a high dose of Cas-Br-M murine leukemia virus, a neuropathogenic type C retrovirus, results in virus-induced neurologic disease and in their failure to generate a protective CTL response. Cas-Br-M-specific CTL are necessary in the protection of neonatal mice from Cas-Br-M-induced neurologic disease. Here we demonstrate that intramuscular inoculation of newborn mice with naked DNA expressing the full length Cas-Br-M genome induces a virus-specific CTL-mediated response. This CTL response is mediated by CD8+ T cells, is long lasting and, when transferred to susceptible neonatal recipients, protects them from Cas-induced neurologic disease. We also provide evidence that the intramuscular inoculation of neonates with plasmid DNA encoding only env sequences induces a dose-dependent CTL response in the absence of an anti-MuLV antibody response.

Ultraviolet-light-inactivated Cas-Br-M murine leukemia virus induces a protective CD8+ cytotoxic T lymphocyte response in newborn mice.

Newborn NFS/N mice are susceptible to the neurological disease induced by infection with Cas-Br-M murine leukemia virus (Cas), and do not develop a protective cytotoxic T cell (CTL)-mediated response to Cas infection. Here we demonstrate that whole UV light-inactivated Cas (UV-Cas), inoculated in newborn NFS/N mice, induced a strong, Cas-specific CTL response detectable 2 weeks postinoculation and persisting in vivo for > or = 36 weeks. The magnitude of the UV-Cas-induced splenic CTL response, mediated by CD8+ T cells, inversely correlated with the level of proviral cas env sequences detectable in the spleen of the UV-Cas-inoculated mice, as revealed by PCR amplification of tissue DNA. The transfer of UV-Cas-primed splenocytes, with Cas-specific CTL activity, protected 100% of recipient newborn mice from the development of neurological disease induced by infection with live Cas, for more than 28 weeks, and reduced the level of viral replication in the recipients.

Preparation of environmental samples for the determination of polycyclic aromatic hydrocarbons by thin-layer chromatography.

An evaluation of extraction procedures, liquid-liquid distribution systems, Sep-Pak cartridges, liquid-solid chromatography using silica, alumina and chemically modified silica packings (acid-base treated ethylammonium nitrate and picric acid impregnated), macroreticular resins and gel permeation columns for the analysis of polycyclic aromatic hydrocarbons (CPAHs) in environmental samples by thin-layer chromatography is discussed. For particulate samples solvent extraction using a Soxhlet apparatus or ultrasonication was found to be preferable to sublimation and liquid-liquid distribution between hexane and dimethyl sulfoxide followed by silica gel column chromatography was the preferred method for sample clean-up. Using this procedure enabled six PAHs (anthracene, fluoranthene, benz[a]anthracene, perylene, pyrene, and coronene) to be determined quantitatively in urban air particulate, diesel engine exhaust particulate, laboratory ventilator dust, household dust, river water, and tea samples. The PAHs were identified by coincidence of retention between the sample and standards in the same chromatographic system and by adequate agreement with standards for their normalized emission response ratios. The two-point calibration method was used for quantitation. Good agreement for the concentration of PAHs in the air particulate and diesel particulate extracts with published data using gas chromatography-mass spectrometry and high-performance liquid chromatography was found.

Development in an estuarine fouling community: The influence of early colonists on later arrivals.

Experiments were performed to determine if earlier colonists inhibited, enhanced, or were necessary for establishment of later colonists during development of an estuarine fouling community at Lewes, Delaware. We determined the significance of earlier stages on the successional process by functionally removing early colonizing species. Since settlement of sessile invertebrates onto our experimental test plates was seasonal, we were able to accomplish functional removal of early colonists by putting out clean test panels after these species had ceased settling. Comparisons between panels initially submerged at three different times in 1974 and 1975, and between panels put out at one-month intervals throughout the study (to describe seasonal settlement patterns) allowed us to determine interactions between adult populations of earlier colonists and colonizing individuals of later arriving species.The dominant sessile species in our system and their times of settlement were: a barnacle (Balanus improvisus) - April through June, a polychaete (Hydroides dianthus) - July and August, a tunicate (Molgula manhatensis) - June through October, a hydroid (Tubularia crocea) - July through October, and a mussel (Mytilus edulis) - November through April. All successional series eventually came to be dominated by M. edulis, and it persisted as the dominant for over a year.A variety of species interactions were observed. M. edulis inhibited colonization by all other dominants and B. improvisus partially inhibited settlement of M. manhattensis. The presence of adult M. manhattensis had no influence on summer settlement of T. crocea, but the hydroids enhanced settlement of tunicates in the fall. During both years of our study, larger settlements of mussels were noted on panels harboring tunicates and hydroids than on bare surfaces. H. dianthus, on the other hand, became established only on bare substrates, and colonization was almost totally inhibited by other dominants.Development in our fouling community did not conform to any single model of community development presented to date. Instead, components of several models were observed within our relatively simple (in terms of number of species) system. For example, facilitation (enhancement of later colonists by earlier ones) and inhibition (resistance of earlier colonists to invasion by later colonists) were both observed. However, we found no evidence earlier colonists were essential for establishment of the next developmental stage. In fact, inhibitory interactions appeared to be much more prevalent than facilitative interactions. The former may also have more profound effects on community development since they more often determine eventual species compositions.