Central effects of ethanol interact with endogenous mu-opioid activity to control isolation-induced analgesia in maternally separated infant rats.

Endogenous opioid activity plays an important role in ethanol consumption and reinforcement in infant rats. Opioid systems are also involved in mediation and regulation of stress responses. Social isolation is a stressful experience for preweanling rats and changes the effects of ethanol through opioid-dependent mechanisms. The present study assessed effects of intracisternal (i.c.) administration of a selective mu-opioid antagonist (CTOP) and i.p. administration of a nonspecific opioid antagonist (naloxone) on voluntary intake and behavior in socially isolated 12-day-old (P12) pups treated with 0.5 g/kg ethanol. Voluntary intake of 0.1% saccharin or water, locomotion, rearing activity, paw licking and grooming were assessed during short-term isolation from littermates (STSI; 8-min duration). Thermal nociceptive reactivity was measured before and after this intake test, with normalized differences between pre- and post-test latencies of paw withdrawal from a hot plate (49°C) used as an index of isolation-induced analgesia (IIA). Results indicated several effects of social isolation and ethanol mediated through the mu-opioid system. Effects of low dose ethanol (0.5 g/kg) and voluntary consumption of saccharin interacted with endogenous mu-opioid activity associated with STSI. Blockade of mu-opioid receptors on saccharin consumption and paw licking-grooming affected intoxicated animals. Low dose ethanol and ingestion of saccharin blunted effects of CTOP on rearing behavior and nociceptive reactivity. Central injections of CTOP stimulated paw licking and grooming dependent on ethanol dose and type of fluid ingested. Ethanol selectively increased saccharin intake during STSI in females, naloxone and CTOP blocked ethanol-mediated enhancement of saccharin intake. We suggest that enhancement of saccharin intake by ethanol during STSI is the product of synergism between isolation-induced mu-opioid activity that increases the pup's sensitivity to appetitive taste stimulation and the anxiolytic effects of 0.5 g/kg ethanol that decreases behaviors otherwise competing with independent ingestive activity.

Access to syringes for HIV prevention for injection drug users in St. Petersburg, Russia: syringe purchase test study.

BACKGROUND: The HIV epidemic in Russia is concentrated among injection drug users (IDUs). This is especially true for St. Petersburg where high HIV incidence persists among the city's estimated 80,000 IDUs. Although sterile syringes are legally available, access for IDUs may be hampered. To explore the feasibility of using pharmacies to expand syringe access and provide other prevention services to IDUs, we investigated the current access to sterile syringes at the pharmacies and the correlation between pharmacy density and HIV prevalence in St. Petersburg. METHODS: 965 pharmacies citywide were mapped, classified by ownership type, and the association between pharmacy density and HIV prevalence at the district level was tested. We selected two districts among the 18 districts--one central and one peripheral--that represented two major types of city districts and contacted all operating pharmacies by phone to inquire if they stocked syringes and obtained details about their stock. Qualitative interviews with 26 IDUs provided data regarding syringe access in pharmacies and were used to formulate hypotheses for the pharmacy syringe purchase test wherein research staff attempted to purchase syringes in all pharmacies in the two districts. RESULTS: No correlation was found between the density of pharmacies and HIV prevalence at the district level. Of 108 operating pharmacies, 38 (35%) did not sell syringes of the types used by IDUs; of these, half stocked but refused to sell syringes to research staff, and the other half did not stock syringes at all. Overall 70 (65%) of the pharmacies did sell syringes; of these, 49 pharmacies sold single syringes without any restrictions and 21 offered packages of ten. CONCLUSIONS: Trainings for pharmacists need to be conducted to reduce negative attitudes towards IDUs and increase pharmacists' willingness to sell syringes. At a structural level, access to safe injection supplies for IDUs could be increased by including syringes in the federal list of mandatory medical products sold by pharmacies.

PBOV1 is a human de novo gene with tumor-specific expression that is associated with a positive clinical outcome of cancer.

PBOV1 is a known human protein-coding gene with an uncharacterized function. We have previously found that PBOV1 lacks orthologs in non-primate genomes and is expressed in a wide range of tumor types. Here we report that PBOV1 protein-coding sequence is human-specific and has originated de novo in the primate evolution through a series of frame-shift and stop codon mutations. We profiled PBOV1 expression in multiple cancer and normal tissue samples and found that it was expressed in 19 out of 34 tumors of various origins but completely lacked expression in any of the normal adult or fetal human tissues. We found that, unlike the cancer/testis antigens that are typically controlled by CpG island-containing promoters, PBOV1 was expressed from a GC-poor TATA-containing promoter which was not influenced by CpG demethylation and was inactive in testis. Our analysis of public microarray data suggests that PBOV1 activation in tumors could be dependent on the Hedgehog signaling pathway. Despite the recent de novo origin and the lack of identifiable functional signatures, a missense SNP in the PBOV1 coding sequence has been previously associated with an increased risk of breast cancer. Using publicly available microarray datasets, we found that high levels of PBOV1 expression in breast cancer and glioma samples were significantly associated with a positive outcome of the cancer disease. We also found that PBOV1 was highly expressed in primary but not in recurrent high-grade gliomas, suggesting the presence of a negative selection against PBOV1-expressing cancer cells. Our findings could contribute to the understanding of the mechanisms behind de novo gene origin and the possible role of tumors in this process.

µ-Opioid blockade reduces ethanol effects on intake and behavior of the infant rat during short-term but not long-term social isolation.

Numerous findings in adult and infant rats have shown that the endogenous opioid system is involved in control of ethanol consumption and its reinforcing effects. Opioid systems are also involved in reactivity to social isolation with several factors (age, duration, and type of isolation) affecting this modulation. The present study investigated the effects of a selective mu-opioid antagonist CTOP (0, 0.1, 0.5mg/kg), ethanol (0, 0.5 g/kg), and the interaction of the two drugs on the behavioral consequences of two types of social isolation given to preweanling rats: 1) short-term social isolation from littermates (STSI, duration 8 min) and 2) relatively long-term (5h) isolation (LTSI) from the dam and littermates. Voluntary intake of saccharin, locomotion, rearing activity, paw licking, and grooming were assessed during an 8-min. intake test. Thermal nociceptive reactivity was also measured before and after the testing session with normalized differences in pre- and post-test latencies of paw withdrawal from a hot plate (49°C) used as an index of isolation-induced analgesia (IIA). The results indicate that pharmacological blockade of mu-opioid receptors by CTOP substantially attenuated ethanol's anxiolytic effects on the developing rat's reactions to social isolation. Some of these stress-attenuating effects of CTOP were observed only in animals exposed to short-term isolation (STSI) but not in pups isolated for 5h (LTSI). Ethanol selectively increased saccharin intake during STSI in females and CTOP blocked this effect. Ethanol decreased the magnitude of analgesia associated with STSI but had no effect on pain reactivity during LTSI. CTOP by itself did not affect IIA or saccharin intake in sober animals. The findings of the present experiments indicate that the anxiolytic effects of 0.5 g/kg ethanol on pups exposed to STSI are modulated by endogenous opioid activity.

The role of social isolation in ethanol effects on the preweanling rat.

The present experiments investigated the effects of acute ethanol exposure on voluntary intake of 0.1% saccharin or water as well as behavioral and nociceptive reactivity in 12-day-old (P12) rats exposed to differing levels of isolation. The effects of ethanol emerged only during short-term social isolation (STSI) with different patterns observed in males and females and in pups exposed to saccharin or water. The 0.5g/kg ethanol dose selectively increased saccharin intake in females, decreased rearing activity in males and attenuated isolation-induced analgesia (IIA) in all water-exposed pups. Ingestion of saccharin decreased IIA, and the 0.5g/kg ethanol dose further reduced IIA. The 1.0g/kg ethanol dose, administered either intragastrically or intraparentionally, also decreased IIA in P12 females, but not in P9 pups. A significant correlation between voluntary saccharin intake and baseline nociceptive reactivity was revealed in saline injected animals, saccharin intake was inversely correlated with behavioral activation and latency of reaction to noxious heat after 0.5g/kg ethanol in females. The 0.5g/kg ethanol dose did not affect plasma corticosterone (CORT) measured 5h after maternal separation or 20min after ethanol injection. Female pups CORT level was inversely correlated with magnitude of IIA that accompanied the first episode of STSI (pretest isolation) 1.5-2h before CORT measurement. The present findings suggest that the anxiolytic properties of ethanol are responsible for enhancement of saccharin intake during STSI. Furthermore, differential reactivity of P12 males and females to STSI plays an important role in ethanol effects observed at this age.

Pharmacological effects of ethanol on ingestive behavior of the preweanling rat.

The present study was designed to test the hypothesis that sensitivity of ingestive behavior of infant rat to the pharmacological effects of ethanol changes between postnatal (P) days 9 and 12. The intake of 0.1% saccharin and water, general motor activity, and myoclonic twitching activity were assessed following administration of three doses of ethanol (0, 0.25, and 0.5 g/kg) while fluids were free available to the animals. The 0.5 g/kg dose of ethanol attenuated saccharin intake in P9 pups and enhanced saccharin intake in P12 rats. On P12 some sex-related differences emerged at 0.5 g/kg of ethanol, with saccharin intake being higher in females than in their male counterparts. Taste reactivity probe revealed that 0.5 g/kg of ethanol increased taste responsiveness to saccharin on P12 but only to infusions presented at a high rate. The results of the present study indicate that ontogenetic changes in sensitivity to the effects of ethanol on ingestive behavior occur during the second postnatal week, with P9 animals being more sensitive to the inhibitory (sedative) effects on saccharin intake and P12 rats being more sensitive to the stimulatory effects of ethanol. We suggest that acute ethanol enhanced saccharin intake via sensitization of oral response to appetitive taste stimulation.

Ethanol, saccharin, and quinine: early ontogeny of taste responsiveness and intake.

BACKGROUND: Rat pups demonstrate high levels of immediate acceptance of ethanol during the first 2 weeks of postnatal life. Given that the taste of ethanol is most likely perceived by infant rats as a combination of sweet and bitter, high intake of ethanol early in ontogeny may be associated with age-related enhanced responsiveness to the sweet component of ethanol taste, as well as with ontogenetic decreases in sensitivity to its bitter component. Therefore, the present study compared responsiveness to ethanol and solutions with bitter (quinine) and sweet (saccharin) taste in terms of intake and palatability across the first 2 weeks of postnatal life. METHODS: Characteristic patterns of responsiveness to 10% (v/v) ethanol, 0.1% saccharin, 0.2% quinine, and water in terms of taste reactivity and fluid intake were assessed in rat pups tested on postnatal day (P) 4, 9, or 12 using a new technique of on-line monitoring of fluid flow through a two-channel intraoral cannula. Taste reactivity included analysis of ingestive and aversive responses following six intraoral infusions of the test fluids. This taste reactivity probe was followed by the intake test, in which animals were allowed to voluntarily ingest fluids from an intraoral cannula. RESULTS: Pups of all ages showed more appetitive responses to saccharin and ethanol than to water or quinine. No age-related differences were apparent in taste responsiveness to saccharin and ethanol. However, the age-related pattern of ethanol intake drastically differed from that of saccharin. Intake of saccharin increased from P4 to P9 and decreased substantially by P12, whereas intake of ethanol gradually increased from P4 to P12. Intake of ethanol was significantly lower than intake of saccharin on P9, whereas P12 pups took in more ethanol than saccharin. CONCLUSION: The findings of the present study indicate ontogenetic dissociations between taste reactivity to ethanol and saccharin and intake of these solutions, and suggest that high acceptance of ethanol early in ontogeny may not be associated with its orosensory properties but rather with the pharmacological effects of ethanol.

Taste differentiation in the context of suckling and independent, adultlike ingestive behavior.

The present study compared intake of sweet (saccharin), bitter (quinine), and neutral (water) tastants available either in the context of suckling behavior through a surrogate nipple or independent adultlike feeding through an intraoral cheek cannula in 3-hr-old newborn rats lacking any suckling experience and 24-hr-old rats with regular experience with the dam's nipple. The new technique of online monitoring of fluid flow was applied for assessment of the temporal patterns of ingestion. Newborn and 1-day-old rats tested in the context of suckling behavior showed extremely low intake of quinine, moderate intake of water, and high intake of saccharin. In the same way, they demonstrated low intake of quinine and high intake of saccharin in the context of independent feeding, but intake of water was also high and comparable to that of saccharin. Suckling rats attained higher efficacy of fluid extraction from nipple than pups drinking from cannula. The differential responsiveness to quinine, saccharin, and water in suckling rats was also manifested through attachment behavior, with pups spending less time on the nipple providing quinine and more time on the nipple with saccharin than on the nipple providing water. These results suggest that neonates show taste differentiation as early as 3 hr after birth, and that this taste differentiation is more pronounced in the context of suckling behavior than in the context of adultlike, independent ingestion.

Repetitive exposures to a surrogate nipple providing nutritive and non-nutritive fluids: effects on suckling behavior of the newborn rat.

Responsiveness to a surrogate nipple providing water, 0.1% saccharin, 10% sucrose, pedialyte, or milk was tested in naïve-to-suckling newborn rats during six 10-min exposures, one every 1.5 h over a 7.5 h period. Across a succession of exposures, newborn rats repeatedly attached to and ingested milk from a surrogate nipple, yielding significant body weight gain and increased concentration of blood plasma glucose. Initially, pups ingested considerable amounts of saccharin and sucrose, but then dramatically decreased their consumption of these fluids across the experimental sessions. Intake of milk was significantly higher than that of all other substances. Blood glucose concentration in pups treated with water, saccharin, sucrose, and pedialyte did not differ significantly from that of non-treated pups. The present data suggest a potential contribution of a fluid's palatability and nutritive value in the persistence and efficacy of diet intake for neonatal rats in the context of suckling behavior.

Oral compression activity on a surrogate nipple in the newborn rat: nutritive and nonnutritive sucking.

Newborn rats, 3 hr after birth and before any experience in suckling, were exposed for 10 min to a surrogate nipple providing milk. One hour later, they were exposed to an empty nipple for another 10-min period. The basic characteristics of oral behavior (oral compression activity, OCA) were assessed by recording intranipple pressure during the pups' first attachment to a nipple. The peculiarities of milk-induced changes of OCA were examined with three modes of milk delivery (milk infusions, and intermittent and continuous milk deliveries). The pattern of OCA exerted by the newborn pups on a surrogate nipple consisted of rhythmic oscillations within a frequency range of 0.4 to 1.0 Hz superimposed on slow (frequency < 0.2 Hz), irregular intranipple pressure fluctuations. Oral behavior during the first minute after oral capture of the nipple differed significantly from that during any subsequent 1-min interval in terms of frequency content of OCA. The pattern of OCA changes induced by milk infusions or intermittent milk delivery included an abrupt rise in intranipple pressure, accompanied or followed by a burst of fast nipple compressions (bites). Our data suggest that newborn rats attached to a surrogate nipple demonstrate patterns of oral behavior that simulate, in terms of basic frequency characteristics, patterns of nutritive and nonnutritive suckling observed in more mature pups on the maternal nipple.