Prior exposure to nutritive and artificial sweeteners differentially alters the magnitude and persistence of sucrose-conditioned flavor preferences in BALB/c and C57BL/6 inbred mouse strains.

Murine genetic variance affects sucrose's ability to condition flavor preferences (CFP) relative to saccharin. Whereas BALB/c mice display robust sucrose- and fructose-CFP, C57BL/6 mice only display sucrose-CFP. Prior exposure to sucrose or saccharin solutions alters subsequent food choice responsiveness. The present study examined whether pre-exposure for one month to 10% sucrose or 0.2% saccharin altered subsequent sucrose-CFP in male and female BALB/c and C57BL/6 mice. Two weeks later, food-restricted mice were exposed to 10 CFP training trials with uniquely flavored 16% sucrose and 0.2% saccharin solutions. Two-bottle choice tests of the flavors mixed in saccharin followed for 4 weeks. Male mice weighed more than females across all conditions, and male BALB/c, but not C57BL mice consumed more 85 sucrose than females. No other notable sex differences were observed. BALB/c mice consumed more sucrose during pre-exposure and one-bottle training than C57BL/6 mice. Although the magnitudes of sucrose-CFP were comparable in two-bottle choice tests in water-exposed BALB/c and C57BL/6 mice, sucrose- and saccharin-exposed BALB/c mice displayed significantly greater sucrose-CFP preferences relative to C57BL/6 counterparts. These data indicate murine genetic variance in the effects of prior exposure to nutritive or non-nutritive sweeteners upon the magnitude of adult sugar-CFP.

Functional germline variants in driver genes of breast cancer.

PURPOSE: Germline mutations in tumour suppressor genes cause various cancers. These genes are also somatically mutated in sporadic tumours. We hypothesized that there may also be cancer-related germline variants in the genes commonly mutated in sporadic breast tumours. METHODS: After excluding the well-characterized breast cancer (BC) genes, we screened 15 novel genes consistently classified as BC driver genes in next-generation sequencing approaches for single nucleotide polymorphisms (SNPs). Altogether 40 SNPs located in the core promoter, 5'- and 3'-UTR or which were nonsynonymous SNPs were genotyped in 782 Swedish incident BC cases and 1,559 matched controls. After statistical analyses, further evaluations related to functional prediction and signatures of selection were performed. RESULTS: TBX3 was associated with BC risk (rs2242442: OR = 0.76, 95% CI 0.64-0.92, dominant model) and with less aggressive tumour characteristics. An association with BC survival and aggressive tumour characteristics was detected for the genes ATR (rs2227928: HR = 1.63; 95% CI 1.00-2.64, dominant model), RUNX1 (rs17227210: HR = 3.50, 95% CI 1.42-8.61, recessive model) and TTN (rs2303838: HR = 2.36; 95% CI 1.04-5.39; rs2042996: HR = 2.28; 95% CI 1.19-4.37, recessive model). According to the experimental ENCODE data all these SNPs themselves or SNPs in high linkage disequilibrium with them (r 2 ≥ 0.80) were located in regulatory regions. RUNX1 and TTN showed also several signatures of positive selection. CONCLUSION: The study gave evidence that germline variants in BC driver genes may have impact on BC risk and/or survival. Future studies could discover further germline variants in known or so far unknown driver genes which contribute to cancer development.

"C.R.E.A.T.E."-ing Unique Primary-Source Research Paper Assignments for a Pleasure and Pain Course Teaching Neuroscientific Principles in a Large General Education Undergraduate Course.

A large (250 registrants) General Education lecture course, Pleasure and Pain, presented basic neuroscience principles as they related to animal and human models of pleasure and pain by weaving basic findings related to food and drug addiction and analgesic states with human studies examining empathy, social neuroscience and neuroeconomics. In its first four years, the course grade was based on weighted scores from two multiple-choice exams and a five-page review of three unique peer-reviewed research articles. Although well-registered and well-received, 18% of the students received Incomplete grades, primarily due to failing to submit the paper that went largely unresolved and eventually resulted in a failing grade. To rectify this issue, a modified version of the C.R.E.A.T.E. (Consider, Read, Elucidate hypotheses, Analyze and interpret data, Think of the next Experiment) method replaced the paper with eight structured assignments focusing on an initial general-topic article, the introduction-methods, and results-discussion of each of three related peer-review neuroscience-related articles, and a final summary. Compliance in completing these assignments was very high, resulting in only 11 INC grades out of 228 students. Thus, use of the C.R.E.A.T.E. method reduced the percentage of problematic INC grades from 18% to 4.8%, a 73% decline, without changing the overall grade distribution. Other analyses suggested the students achieved a deeper understanding of the scientific process using the C.R.E.A.T.E. method relative to the original term paper assignment.

Muscarinic and nicotinic cholinergic receptor antagonists differentially mediate acquisition of fructose-conditioned flavor preference and quinine-conditioned flavor avoidance in rats.

Rats display both conditioned flavor preference (CFP) for fructose, and conditioned flavor avoidance (CFA) following sweet adulteration with quinine. Previous pharmacological analyses revealed that fructose-CFP expression was significantly reduced by dopamine (DA) D1 or D2 antagonists, but not NMDA or opioid antagonists. Fructose-CFP acquisition was significantly reduced by DA D1, DA D2 or NMDA antagonists, but not opioid antagonists. Quinine-CFA acquisition was significantly enhanced and prolonged by DA D1, NMDA or opioid, but not DA D2 antagonists. Cholinergic interneurons and projections interact with DA systems in the nucleus accumbens and ventral tegmental area. Further, both muscarinic and nicotinic cholinergic receptor signaling have been implicated in sweet intake and development of food-related preferences. Therefore, the present study examined whether systemic administration of muscarinic (scopolamine: SCOP) or nicotinic (mecamylamine: MEC) cholinergic receptor antagonists mediated fructose-CFP expression, fructose-CFP acquisition and quinine-CFA acquisition. For fructose-CFP expression, rats were trained over 10 sessions with a CS+ flavor in 8% fructose and 0.2% saccharin and a CS- flavor in 0.2% saccharin. Two-bottle choice tests with CS+ and CS- flavors mixed in 0.2% saccharin occurred following vehicle, SCOP (0.1-10mg/kg) and MEC (1-8mg/kg). For fructose-CFP acquisition, six groups of rats received vehicle, SCOP (1 or 2.5mg/kg), MEC (4 or 6mg/kg) or a limited intake vehicle control 0.5h prior to 10 CS+ and CS- training sessions followed by six 2-bottle CS+ and CS- choice tests in 0.2% saccharin. For quinine-CFA acquisition, five groups of rats received vehicle, SCOP (1 or 2.5mg/kg) or MEC (4 or 6mg/kg) 0.5h prior to 8 one-bottle CS- (8% fructose+0.2% saccharin: FS) and CS+ (fructose+saccharin+quinine (0.030%: FSQ) training sessions followed by six 2-bottle CS- and CS+ choice tests in fructose-saccharin solutions. Fructose-CFP expression was significantly reduced by SCOP (2.5-10mg/kg: 65-68%) and MEC (4-8mg/kg: 67-73%) relative to vehicle (89-90%), that occurred only when antagonist doses reduced total saccharin intake but in which CS+ intake was still significantly higher than CS- intake. Fructose-CFP acquisition was eliminated by SCOP at doses of 1 (40-54%) and 2.5 (45-58%)mg/kg, and was accompanied by a failure to observe CS+ and CS- intake differences during testing relative to vehicle (85-92%) and limited control (74-88%) conditions. In contrast, MEC failed to alter fructose-CFP acquisition. Quinine-CFA acquisition was significantly enhanced and prolonged by MEC at 4 (18-24%) and 6 (11-13%) mg/kg relative to vehicle (34-48%). In contrast, SCOP failed to alter quinine-CFA acquisition. These data implicate the cholinergic receptor system in mediating acquisition (learning) of sugar-induced preferences and quinine-induced aversions with muscarinic receptor signaling controlling the former and nicotinic receptor signaling controlling the latter.

Intraoperative changes in blood pressure, heart rate, plasma vasopressin, and urinary noradrenalin during elective ovariohysterectomy in dogs: repeatability at removal of the 1st and 2nd ovary.

OBJECTIVE: To investigate the physiologic reactions after removal of 1st ovary and whether this is repeated during removal of the 2nd ovary in elective ovariohysterectomy. STUDY DESIGN: Prospective study. ANIMALS: Dogs (n = 10). METHODS: Dogs were premedicated with acepromazine, carprofen, and methadone and anesthetized with propofol and isoflurane. Blood pressure, heart rate, and end-tidal isoflurane concentration were measured every minute. The effects of various events during surgery on physiologic variables were analyzed using mixed linear models. Blood and urine samples were collected before anesthesia, before incision, before and after removal of ovaries with a 15 minute pause between ovary removal, and after abdominal closure. Plasma vasopressin and urinary noradrenalin and creatinine concentrations were analyzed. RESULTS: The magnitude of blood pressure increase at removal of the 1st ovary was greater than for the 2nd ovary because of an elevation in baseline. Similarly, the heart rate increased at the removal of the 1st ovary but not at removal of the 2nd ovary. Plasma vasopressin concentration increased at removal of both ovaries. Urinary noradrenalin/creatinine ratio increased at anesthesia, removal of both ovaries, and was elevated at closure of the abdomen. End-tidal isoflurane concentration did not change. Blood pressure and vasopressin concentrations changed in parallel using z-scores for comparison. CONCLUSIONS: Peak values for blood pressure, heart rate, plasma vasopressin concentration, and urinary noradrenalin/creatinine ratio did not differ between removals of the ovaries. Relative changes differed between repeated noxious stimuli, which should be considered in evaluation of methods at ovary removal.

Physiological and behavioral responses to different watering intervals in lactating camels (Camelus dromedarius).

During drought periods camels are watered at long intervals, but effects on body fluid homeostasis of lactating camels are not known. It was hypothesized that camels store water after drinking and minimize water losses by diurnal variation in body temperature, changes in behavior, and release of vasopressin. The aim was to find a sustainable watering interval for lactating camels. Seven lactating camels were studied in a cross-over trial in which they were watered once daily (W1), every fourth day (W4), every eighth day (W8), or after 16 days (W16) with a 5-day interval between treatments. When offered water every fourth or eighth days, the camels drank sufficient amounts to cover their needs for subsequent days, but after 16 days of dehydration they did not drink enough to compensate the body weight loss. Rectal temperature fell at night and the camels searched shade during daytime minimizing evaporative fluid losses. Plasma osmolality and sodium concentration were elevated after 4 days of water deprivation and plasma protein and vasopressin concentrations after 8 days. Milk production decreased during the last week of W16. Plasma aldosterone concentration was elevated upon rehydration after W16, indicating sodium deficiency. In conclusion, lactating camels stored water after drinking and reduced water losses by staying in shade, keeping body temperature low, and releasing plasma vasopressin. However, serious dehydration was observed during W8, and after 16 days of water deprivation recovery took a long time. A watering interval between 4 and 7 days seems advisable under similar environmental conditions.

The group rehabilitation helped me adjust to a new life: experiences shared by persons with an acquired brain injury.

PRIMARY OBJECTIVE: The aim of this study was to describe how persons with acquired brain injury experience an out-patient group rehabilitation programme and how the programme had contributed to their everyday lives. DESIGN AND METHOD: Qualitative interviews with 11 men and women with an acquired brain injury who had participated in an out-patient group rehabilitation programme were performed. Data was analysed with qualitative content analysis. FINDINGS: The findings formed the theme 'The group rehabilitation helped me adjust to a new life' that revealed experiences related to the content and outcome of the programme, as well as the process they went through during the programme. The participants described how the rehabilitation gave them the tools they needed to change their everyday lives, especially in relation to improved knowledge and learning new routines and habits. They perceived their rehabilitation as a long-term, individual, but also collaborative process, where professionals as well as family and friends had crucial roles. CONCLUSION: Learning how persons with acquired brain injury experience participation in a group rehabilitation programme can help to unravel parts of the 'black box of rehabilitation' and can support professionals to better understand the effective components of such programmes.

Pleasure and pain: teaching neuroscientific principles of hedonism in a large general education undergraduate course.

In a large (250 registrants) general education lecture course, neuroscience principles were taught by two professors as co-instructors, starting with simple brain anatomy, chemistry, and function, proceeding to basic brain circuits of pleasure and pain, and progressing with fellow expert professors covering relevant philosophical, artistic, marketing, and anthropological issues. With this as a base, the course wove between fields of high relevance to psychology and neuroscience, such as food addiction and preferences, drug seeking and craving, analgesic pain-inhibitory systems activated by opiates and stress, neuroeconomics, unconscious decision-making, empathy, and modern neuroscientific techniques (functional magnetic resonance imaging and event-related potentials) presented by the co-instructors and other Psychology professors. With no formal assigned textbook, all lectures were PowerPoint-based, containing links to supplemental public-domain material. PowerPoints were available on Blackboard several days before the lecture. All lectures were also video-recorded and posted that evening. The course had a Facebook page for after-class conversation and one of the co-instructors communicated directly with students on Twitter in real time during lecture to provide momentary clarification and comment. In addition to graduate student Teaching Assistants (TAs), to allow for small group discussion, ten undergraduate students who performed well in a previous class were selected to serve as discussion leaders. The Discussion Leaders met four times at strategic points over the semester with groups of 20-25 current students, and received one credit of Independent Study, thus creating a course within a course. The course grade was based on weighted scores from two multiple-choice exams and a five-page writing assignment in which each student reviewed three unique, but brief original peer-review research articles (one page each) combined with expository writing on the first and last pages. A draft of the first page, collected early in the term, was returned to each student by graduate TAs to provide individual feedback on scientific writing. Overall the course has run three times at ful or near enrollment capacity despite being held at an 8:00 AM time slot. Student-generated teaching evaluations place it well within the normal range, while this format importantly contributes to budget efficiency permitting the teaching of more required small-format courses (e.g., freshman writing). The demographics of the course have changed to one in which the vast majority of the students are now outside the disciplines of neuroscience or psychology and are taking the course to fulfill a General Education requirement. This pattern allows the wide dissemination of basic neuroscientific knowledge to a general college audience.

Prognostic impact of polymorphisms in the MYBL2 interacting genes in breast cancer.

MYBL2 is a transcription factor, which regulates the expression of genes involved in cancer progression. In this study, we investigated whether putative functional variants in genes regulating MYBL2 (E2F1, E2F3 and E2F4) or in genes, which are regulated by MYBL2 (BCL2, BIRC5, COL1A1, COL1A2, COL5A2, ERBB2, CLU, LIN9 and TOP2A) affect breast cancer (BC) susceptibility and clinical outcome. Twenty-eight SNPs were genotyped in a population-based series of 782 Swedish BC cases and 1,559 matched controls. BC-specific survival analysis of BIRC5 suggested that carriers of the minor allele of rs8073069 and rs1042489 have a worse survival compared with the major homozygotes (HR 2.46, 95% CI 1.39-4.36 and HR 1.81, 95% CI 1.01-3.25, respectively). The poor survival was observed especially in women with aggressive tumours. Multivariate analysis supported the role of rs8073069 as an independent prognostic marker. For BCL2, minor allele carriers of rs1564483 were more likely to have hormone receptor-positive tumours than the major homozygotes. Another SNP in BCL2, rs4987852, was associated with tumour stages II-IV and histologic grade 3. In CLU, the minor allele carriers of rs9331888 were more likely to have tumours with regional lymph node metastasis and stages II-IV than the major homozygotes. In conclusion, our study suggests a role of genetic variation in BIRC5, BCL2 and CLU as progression and prognostic markers for BC, supporting previous studies based on the expression of the genes.

Genetic variation in ALCAM and other chromosomal instability genes in breast cancer survival.

Chromosomal instability is a hallmark of many cancers and it has a potential to predict clinical outcome of a cancer patient. We hypothesized that genes whose expression status differs between chromosomal stable and unstable breast tumors represent target genes for the identification of genetic variants predicting breast cancer (BC) risk, disease progression, and survival. We used a published list of 38 genes associated with chromosomal instability as a basis for searching potentially functional and informative tagging single nucleotide polymorphisms (SNPs). As a result, 33 SNPs in 16 genes were genotyped in a population-based series of 783 Swedish BC cases. Two SNPs in the ALCAM gene associated with BC-specific survival. For rs1044243, the HR was 4.35 (95% CI 1.34-14.18), and for rs1157, the HR was 3.42 (95% CI 1.32-8.83) for the homozygous carriers of the minor alleles. For the minor allele carriers of CCL18 SNP rs14304, we observed a significant association with aggressive tumor characteristics: large tumor size (OR 1.53, 95% CI 1.10-2.14), positive lymph node metastasis (OR 1.75, 95% CI 1.02-3.00), and high stage (OR 1.37, 95% CI 1.02-1.85). In a Polish population consisting of 506 familial/early onset BC cases, no association with event-free survival for the ALCAM SNPs nor any association with tumor characteristics for the CCL18 SNP were observed, suggesting either a chance finding in the Swedish population or population-based or etiological differences between sporadic and familial/early onset BC.

Clinical pharmacology of methadone in dogs.

OBJECTIVE: To investigate the pharmacokinetics and effects of methadone on behaviour and plasma concentrations of cortisol and vasopressin in healthy dogs. STUDY DESIGN: Randomized, cross-over, experimental trial. ANIMALS: Nine adult dogs (beagle and beagle cross breeds), four males and five females. METHODS: Methadone hydrochloride, 0.4 mg kg(-1), was administered intravenously (IV) and subcutaneously (SC) with a crossover design. Drug and hormone analyses in plasma were performed using Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry and radioimmunoassay respectively. Behavioural data were collected using a standardized protocol. RESULTS: After IV administration, the plasma concentration of methadone at 10 minutes was 82.1 +/- 9.2 ng mL(-1) (mean +/- SD), the terminal half-life was 3.9 +/- 1.0 hours, the volume of distribution 9.2 +/- 3.3 L kg(-1) and plasma clearance 27.9 +/- 7.6 mL minute(-1) kg(-1). After SC administration, time to maximal plasma concentration was 1.26 +/- 1.04 hours and maximal plasma concentration of methadone was 23.9 +/- 14.4 ng mL(-1), the terminal half-life was 10.7 +/- 4.3 hours and bioavailability was 79 +/- 22%. Concentrations of both cortisol and vasopressin were increased for an hour following IV methadone. The observed behavioural effects of methadone were decreased licking and swallowing and an increase in whining after SC administration. The latter finding is notable as it can be misinterpreted as pain when methadone is used as an analgesic. CONCLUSION AND CLINICAL RELEVANCE: When methadone was administered by the SC route, the half-life was longer, but the individual variation in plasma concentrations was greater compared with IV administration. Increased frequency of whining occurred after administration of methadone and may be a drug effect and not a sign of pain. Cortisol and vasopressin concentrations in plasma may not be suitable for evaluating analgesia after methadone treatment.

Plasma vasopressin, oxytocin, estradiol, and progesterone related to water and sodium excretion in normal pregnancy and gestational hypertension.

OBJECTIVE: To investigate associations between plasma oxytocin and vasopressin concentrations and renal water and sodium excretion during normal pregnancy in comparison with gestational hypertension. DESIGN: A prospective open trial conducted in the 12th, 24th, and 36th weeks of gestation. SETTINGS: Seven antenatal clinics in Sweden. PARTICIPANTS: Thirty-seven normotensive women, 15 women with gestational hypertension, and five women with mild preeclampsia. MAIN OUTCOME MEASURES: Hormones were analyzed with radioimmunoassay. Albumin, osmolality, sodium, and urea were analyzed by routine methods. RESULTS: Blood pressure was elevated in the hypertensive women and body mass index in mild preeclampsia from week 12. Renal sodium excretion did not differ between groups or weeks and mean renal free water clearance was negative. In normotensive women, the vasopressin concentration was 1.1+/-0.2 (week 12) and 0.7+/-0.1 pmol/L (week 36: p = 0.053). In hypertensive women, vasopressin concentration was 1.7+/-1.0 pmol/L, week 12, and 0.7+/-0.1 pmol/L in week 36 (ns). In normotensive women, oxytocin concentration increased from 23+/-1 pmol/L in week 12 to 48+/-3 pmol/L in week 36 (p<0.001). Corresponding values in hypertensive women were 36+/-11 (week 12) and 55+/-5 pmol/L (week 36: ns). In all groups, plasma estradiol concentration increased. Plasma progesterone increased until week 24 in normotensive and hypertensive women with further increase in normotensive women. CONCLUSIONS: The low plasma vasopressin and increasing plasma oxytocin concentrations with unchanged water and sodium excretion indicate that oxytocin assists vasopressin in concentrating urine during pregnancy.

Murine genetic variance in muscarinic cholinergic receptor antagonism of sucrose and saccharin solution intakes in three inbred mouse strains.

Nutritive (e.g., sucrose) and non-nutritive (e.g., saccharin) sweeteners stimulate intake in inbred mouse strains. BALB/c, SWR and C57BL/6 mice differ in the ability of dopamine (DA) D1 (SCH23390) and opioid (naltrexone) receptor antagonism to alter sucrose intake. Whereas SCH23390 comparably reduced cumulative sucrose intake in all three strains, naltrexone reduced cumulative sucrose intake maximally in C57/BL/6 mice, in intermediate fashion in BALB/c mice, but not in SWR mice. Whereas cumulative saccharin intake was reduced by DA D1 receptor antagonism in BALB/c and SWR mice, naltrexone was more potent in SWR relative to BALB/c mice. The present study first examined whether SCH23390 (50-1600nmol/kg) and naltrexone (0.01-5mg/kg) altered saccharin intake in C57BL/6 mice. Given that scopolamine (SCOP), a muscarinic cholinergic receptor antagonist, reduces sweet intake in outbred rats, a second experiment examined whether SCOP (0.1-10mg/kg) altered 0.2% saccharin and 10% sucrose intakes in BALB/c, SWR and C57BL/6 mice. Cumulative saccharin intake was significantly reduced by SCH23390 (200-1600nmol/kg; ID40=175nmol/kg) and naltrexone (0.1-5mg/kg; ID40>5mg/kg) in C57BL/6 mice. Cumulative sucrose intake was significantly reduced following SCOP in C57BL/6 (0.1-10mg/kg; ID40=2.32mg/kg) and BALB/c (2.5-10mg/kg; ID40=0.52mg/kg) mice. In contrast, SWR mice (ID40=41.61mg/kg) only displayed transient (15min) reductions in sucrose intake following SCOP (2.5-10mg/kg). Cumulative saccharin intake was significantly reduced following SCOP in C57BL/6 and BALB/c mice (0.1-10mg/kg; ID40<0.1mg/kg). In contrast, SWR mice (ID40=2.28mg/kg) displayed smaller significant reductions in saccharin intake following SCOP (0.1-10mg/kg). These data indicate that although both nutritive and non-nutritive sweet intakes are governed by muscarinic cholinergic receptor signaling, this process is subject to murine genetic variance with greater sensitivity observed in C57BL/6 and BALB/c relative to SWR inbred mouse strains.

Baclofen differentially mediates fructose-conditioned flavor preference and quinine-conditioned flavor avoidance in rats.

Rats display both fructose-conditioned flavor preference (CFP) and quinine conditioned flavor avoidance (CFA). Dopamine (D1 and D2), muscarinic and nicotinic, but not NMDA or opioid receptor antagonists reduced fructose-CFP expression. Dopamine D1, dopamine D2, muscarinic or NMDA, but not opioid or nicotinic receptor antagonists reduced fructose-CFP acquisition. Dopamine D1, NMDA, nicotinic or opioid, but not dopamine D2 or muscarinic receptor antagonists enhanced quinine-CFA acquisition. Baclofen (BAC), a GABAB receptor agonist, alternately enhances or reduces feeding under specific conditions. The present study examined whether systemic BAC administration mediated fructose-CFP expression and acquisition or quinine-CFA acquisition. Fructose-CFP expression studies trained rats with one flavor (CS+) in 8% fructose and 0.2% saccharin and a second (CS-) flavor in 0.2% saccharin, followed by vehicle (VEH) and BAC (0.5-5 mg/kg) preceding 2-bottle (CS+, CS-) 0.2% saccharin choice tests. Fructose-CFP acquisition studies administered VEH or BAC (3 or 5 mg/kg) prior to CS+ and CS- training sessions followed by six 2-bottle (CS+, CS-) 0.2% saccharin choice tests. Quinine-CFA acquisition studies administered VEH or BAC (3 or 5 mg/kg) prior to CS- (8% fructose+0.2% saccharin) and CS+ (fructose+saccharin+0.030% quinine) training sessions followed by six 2-bottle (CS-, CS+) fructose+saccharin choice tests. BAC (3 mg/kg) minimally (66%) reduced fructose-CFP expression. BAC failed to alter fructose-CFP acquisition. Quinine-CFA acquisition was enhanced by the 5 mg/kg BAC dose (15-25%) relative to VEH (34-48%). These data implicate GABAB receptor signaling in acquisition of quinine avoidance with minimal or no effects upon fructose preferences.

Muscarinic, nicotinic and GABAergic receptor signaling differentially mediate fat-conditioned flavor preferences in rats.

Rats display conditioned flavor preferences (CFP) for fats. Previous studies demonstrated that whereas expression of an already-acquired corn oil (CO)-CFP was mildly reduced by dopamine (DA) D1, DA D2, NMDA or opioid receptor antagonists, the acquisition or learning of CO-CFP was eliminated by NMDA antagonists, and significantly reduced by DA D1 and D2, but not opioid antagonists. Previous studies of fructose-CFP demonstrated that muscarinic (scopolamine) and nicotinic (mecamylamine) cholinergic receptor antagonists and GABAB (baclofen) receptor agonism reduced the expression of this acquired response, and that scopolamine, but not mecamylamine or baclofen eliminated the acquisition or learning of this response. The present study examined scopolamine, mecamylamine or baclofen effects upon expression or acquisition of CO-CFP. For expression, rats were trained over 10 sessions with CS+ (3.5% CO) and CS- (0.9% CO) flavored solutions without drugs. Two-bottle choice tests with CS+ and CS- flavors in 0.9% CO examined preferences following vehicle, scopolamine (1-10mg/kg), mecamylamine (1-8mg/kg) and baclofen (1.5-5mg/kg). In acquisition, eight groups of rats received vehicle, scopolamine (1, 2.5mg/kg), mecamylamine (4, 6mg/kg), baclofen (3, 5mg/kg) or a limited intake vehicle control 0.5h prior to all 10 CS+ and CS- training sessions followed by six 2-bottle CS+ and CS- choice tests in 0.9% CO. CO-CFP expression (percent CS+ intake) was significantly but marginally reduced by scopolamine (70%), mecamylamine (85%) and baclofen (74%) relative to vehicle (98%). CO-CFP acquisition was eliminated (41%) by scopolamine relative to vehicle (88%) and limited control (98%) conditions. Neither mecamylamine nor baclofen altered CO-CFP acquisition. Thus, the muscarinic cholinergic receptor system is essential for acquisition (learning) of both fat-induced and sugar (fructose)-induced preferences. In contrast, muscarinic, nicotinic and GABAB receptors were minimally involved in the expression (maintenance) of fat- and fructose-CFP.

Impact of functional germline variants and a deletion polymorphism in APOBEC3A and APOBEC3B on breast cancer risk and survival in a Swedish study population.

PURPOSE: The C → T mutation signature caused by APOBEC family members contributes to the development of breast cancer (BC). Also overexpression of APOBEC3B and a ~29.5-kb deletion polymorphism between APOBEC3A and APOBEC3B have been associated with increased BC risk. METHODS: We investigated in a population-based study, with 782 Swedish BC cases and 1559 controls, associations between potentially functional germline variants in APOBEC3A or APOBEC3B gene and BC risk and survival. Additionally, we identified deletion polymorphism carriers and explored possible associations with BC. RESULTS: No evidence of association between any germline variant, including the deletion polymorphism, and BC risk or survival was observed. Only APOBEC3A promoter polymorphism rs5757402 was associated with low stage (OR = 0.69, 95 % CI 0.50-0.96, dominant model). CONCLUSION: The reported association between the deletion polymorphism and BC risk was not confirmed in the Swedish population, nor did any genotyped germline variant show any association with BC risk or survival.

Fluid balance in ruminants: adaptation to external and internal challenges.

Ruminants are widespread in hot, arid regions. This demands adaptation to large circadian temperature fluctuations and recurrent periods of food and water shortage. Pregnancy and lactation add to the demands on the adaptive mechanisms due to the greater need for food, water, and electrolytes. The blood volume increases to meet the requirements of the fetoplacental unit and the mammary glands. Unlike urine, the milk cannot be concentrated by antidiuretic hormone (vasopressin). During water deprivation, lactating animals therefore become dehydrated more rapidly than nonlactating animals. Nevertheless, desert-adapted lactating ruminants endure frequent periods of water deprivation without incurring bad health. For the offspring living in hot and dry conditions, it is an advantage that the milk is not concentrated, even if the mother has a high antidiuretic hormone concentration to enable her to concentrate the urine. Since ruminants are prey, they need to drink rapidly when they get access to water. The forestomach allows the animals to store water in the reticulorumen. There is no danger of water intoxication even if they drink to satisfaction in a couple of minutes after having lost as much as 30% of their body weight.

Dopamine receptor signaling in the medial orbital frontal cortex and the acquisition and expression of fructose-conditioned flavor preferences in rats.

Systemic dopamine (DA) D1 (SCH23390: SCH) and D2 (raclopride: RAC) antagonists blocked fructose-conditioned flavor preference (CFP) acquisition and expression. Fructose-CFP acquisition was eliminated by medial prefrontal cortex (mPFC) SCH and mPFC or amygdala (AMY) RAC. Fructose-CFP expression was reduced following SCH or RAC in AMY or nucleus accumbens (NAc). The present study examined fructose-CFP acquisition and expression following SCH and RAC in the medial orbital frontal cortex (MOFC), another ventral tegmental area DA target. For fructose-CFP acquisition, five groups of rats received vehicle, SCH (24 or 48 nmol) or RAC (24 or 48 nmol) in the MOFC 0.5h prior to 8 training sessions with one flavor (CS+/Fs) mixed in 8% fructose and 0.2% saccharin, and another flavor (CS-/s) mixed in 0.2% saccharin. In six 2-bottle choice tests in 0.2% saccharin, similar fructose-CFP preferences occurred in groups trained with vehicle (76-77%), SCH24 (69-78%), SCH48 (70-74%) and RAC48 (85-92%). RAC24-trained rats displayed significant CS+ preferences during the first (79%) and third (71%), but not second (58%) test pair. For fructose-CFP expression, rats similarly trained with CS+/Fs and CS- solutions received 2-bottle choice tests following MOFC injections of SCH or RAC (12-48 nmol). CS+ preference expression was significantly reduced by RAC (48 nmol: 58%), but not SCH relative to vehicle (78%). A control group receiving RAC in the dorsolateral prefrontal cortex displayed fructose-CFP expression similar to vehicle. These data demonstrate differential frontal cortical DA mediation of fructose-CFP with mPFC D1 and D2 signaling exclusively mediating acquisition, and MOFC D2 signaling primarily mediating expression.

Inbreeding and homozygosity in breast cancer survival.

Genome-wide association studies (GWASs) help to understand the effects of single nucleotide polymorphisms (SNPs) on breast cancer (BC) progression and survival. We performed multiple analyses on data from a previously conducted GWAS for the influence of individual SNPs, runs of homozygosity (ROHs) and inbreeding on BC survival. (I.) The association of individual SNPs indicated no differences in the proportions of homozygous individuals among short-time survivors (STSs) and long-time survivors (LTSs). (II.) The analysis revealed differences among the populations for the number of ROHs per person and the total and average length of ROHs per person and among LTSs and STSs for the number of ROHs per person. (III.) Common ROHs at particular genomic positions were nominally more frequent among LTSs than in STSs. Common ROHs showed significant evidence for natural selection (iHS, Tajima's D, Fay-Wu's H). Most regions could be linked to genes related to BC progression or treatment. (IV.) Results were supported by a higher level of inbreeding among LTSs. Our results showed that an increased level of homozygosity may result in a preference of individuals during BC treatment. Although common ROHs were short, variants within ROHs might favor survival of BC and may function in a recessive manner.

From wild wolf to domestic dog: gene expression changes in the brain.

Despite the relatively recent divergence time between domestic dogs (Canis familiaris) and gray wolves (Canis lupus), the two species show remarkable behavioral differences. Since dogs and wolves are nearly identical at the level of DNA sequence, we hypothesize that the two species may differ in patterns of gene expression. We compare gene expression patterns in dogs, wolves and a close relative, the coyote (Canis latrans), in three parts of the brain: hypothalamus, amygdala and frontal cortex, with microarray technology. Additionally, we identify genes with region-specific expression patterns in all three species. Among the wild canids, the hypothalamus has a highly conserved expression profile. This contrasts with a marked divergence in domestic dogs. Real-time PCR experiments confirm the altered expression of two neuropeptides, CALCB and NPY. Our results suggest that strong selection on dogs for behavior during domestication may have resulted in modifications of mRNA expression patterns in a few hypothalamic genes with multiple functions. This study indicates that rapid changes in brain gene expression may not be exclusive to the development of human brains. Instead, they may provide a common mechanism for rapid adaptive changes during speciation, particularly in cases that present strong selective pressures on behavioral characters.