The effects of electronic nicotine vapor on voluntary alcohol consumption in female and male C57BL/6 J mice.

SIGNIFICANCE: Alcohol drinking and nicotine vaping often co-occur and dependence on both substances is common. However, the impact of nicotine vaping on alcohol consumption is not fully understood. METHODS: We examined the effects of nicotine vaping on ethanol drinking in female and male C57BL/6 J mice using an electronic nicotine delivery system and intermittent access two-bottle choice (IA-2BC) drinking. Mice were exposed to electronic nicotine vapor (3%) or propylene glycol/vegetable glycerol (PG/VG) control for 3 h sessions daily for 4 weeks and voluntary alcohol consumption was monitored. Nicotine vapor exposure was stopped and voluntary alcohol drinking was measured for a 2 week abstinence period. We also examined the effects of alcohol and nicotine on locomotion, temperature, and nicotine metabolism. RESULTS: Following acute nicotine vapor exposure, alcohol drinking was increased in males but not in females. Thermoregulation was disrupted following nicotine vapor exposure and voluntary drinking. Male and female mice displayed increased locomotor activity immediately following chronic nicotine vapor exposure, and an anxiolytic effect was seen in males. In nicotine vapor abstinence, female mice displayed increased alcohol consumption. Locomotor activity and anxiolytic effects remained elevated in male but not female mice. Female mice displayed higher levels of serum nicotine and hydroxycotinine, suggesting impaired metabolism following chronic drinking and nicotine vapor exposure. CONCLUSION: Collectively, these results suggest that while both male and female ethanol-drinking mice experience the stimulatory effects of nicotine vapor, only in males is there a parallel increase in ethanol drinking and only females display impairments in nicotine metabolism after drinking.

A bright γ-ray flare interpreted as a giant magnetar flare in NGC 253.

Soft γ-ray repeaters exhibit bursting emission in hard X-rays and soft γ-rays. During the active phase, they emit random short (milliseconds to several seconds long), hard-X-ray bursts, with peak luminosities1 of 1036 to 1043 erg per second. Occasionally, a giant flare with an energy of around 1044 to 1046 erg is emitted2. These phenomena are thought to arise from neutron stars with extremely high magnetic fields (1014 to 1015 gauss), called magnetars1,3,4. A portion of the second-long initial pulse of a giant flare in some respects mimics short γ-ray bursts5,6, which have recently been identified as resulting from the merger of two neutron stars accompanied by gravitational-wave emission7. Two γ-ray bursts, GRB 051103 and GRB 070201, have been associated with giant flares2,8-11. Here we report observations of the γ-ray burst GRB 200415A, which we localized to a 20-square-arcmin region of the starburst galaxy NGC 253, located about 3.5 million parsecs away. The burst had a sharp, millisecond-scale hard spectrum in the initial pulse, which was followed by steady fading and softening over 0.2 seconds. The energy released (roughly 1.3 × 1046 erg) is similar to that of the superflare5,12,13 from the Galactic soft γ-ray repeater SGR 1806-20 (roughly 2.3 × 1046 erg). We argue that GRB 200415A is a giant flare from a magnetar in NGC 253.

Rapid spectral variability of a giant flare from a magnetar in NGC 253.

Magnetars are neutron stars with extremely strong magnetic fields (1013 to 1015 gauss)1,2, which episodically emit X-ray bursts approximately 100 milliseconds long and with energies of 1040 to 1041 erg. Occasionally, they also produce extremely bright and energetic giant flares, which begin with a short (roughly 0.2 seconds), intense flash, followed by fainter, longer-lasting emission that is modulated by the spin period of the magnetar3,4 (typically 2 to 12 seconds). Over the past 40 years, only three such flares have been observed in our local group of galaxies3-6, and in all cases the extreme intensity of the flares caused the detectors to saturate. It has been proposed that extragalactic giant flares are probably a subset7-11 of short γ-ray bursts, given that the sensitivity of current instrumentation prevents us from detecting the pulsating tail, whereas the initial bright flash is readily observable out to distances of around 10 to 20 million parsecs. Here we report X-ray and γ-ray observations of the γ-ray burst GRB 200415A, which has a rapid onset, very fast time variability, flat spectra and substantial sub-millisecond spectral evolution. These attributes match well with those expected for a giant flare from an extragalactic magnetar12, given that GRB 200415A is directionally associated13 with the galaxy NGC 253 (roughly 3.5 million parsecs away). The detection of three-megaelectronvolt photons provides evidence for the relativistic motion of the emitting plasma. Radiation from such rapidly moving gas around a rotating magnetar may have generated the rapid spectral evolution that we observe.

Observation of inverse Compton emission from a long γ-ray burst.

Long-duration γ-ray bursts (GRBs) originate from ultra-relativistic jets launched from the collapsing cores of dying massive stars. They are characterized by an initial phase of bright and highly variable radiation in the kiloelectronvolt-to-megaelectronvolt band, which is probably produced within the jet and lasts from milliseconds to minutes, known as the prompt emission1,2. Subsequently, the interaction of the jet with the surrounding medium generates shock waves that are responsible for the afterglow emission, which lasts from days to months and occurs over a broad energy range from the radio to the gigaelectronvolt bands1-6. The afterglow emission is generally well explained as synchrotron radiation emitted by electrons accelerated by the external shock7-9. Recently, intense long-lasting emission between 0.2 and 1 teraelectronvolts was observed from GRB 190114C10,11. Here we report multi-frequency observations of GRB 190114C, and study the evolution in time of the GRB emission across 17 orders of magnitude in energy, from 5 × 10-6 to 1012 electronvolts. We find that the broadband spectral energy distribution is double-peaked, with the teraelectronvolt emission constituting a distinct spectral component with power comparable to the synchrotron component. This component is associated with the afterglow and is satisfactorily explained by inverse Compton up-scattering of synchrotron photons by high-energy electrons. We find that the conditions required to account for the observed teraelectronvolt component are typical for GRBs, supporting the possibility that inverse Compton emission is commonly produced in GRBs.

Together Eating & Activity Matters (TEAM): results of a pilot randomized-clinical trial of a spousal support weight loss intervention for Black men.

Objectives: The objective of this study is to evaluate the feasibility, participation, preliminary efficacy and retention in a couples-based weight loss intervention among Black men. Design setting participants: Two-arm pilot randomized clinical trial in an academic clinical setting. Forty self-identified Black men and their female cohabitating partners (n = 80) aged 18 to 65 years with body mass index from 25 to 45 kg/m2 were randomized using computer generated tables to allocate treatments. Intervention: Participants were randomized to a standard behavioural weight loss (Standard) programme or the Standard programme plus partner involvement (Enhanced). Both interventions focused on calorie reduction, physical activity and self-monitoring to facilitate weight loss. Enhanced included couples skills training and couple's communication components. Main outcome and measures: Changes in weight from baseline to 3 months among men. Partner weight loss (secondary). Results: Forty Black couples (men mean [SD] age, 47.4[11] years; body mass index, 35.0[6.1]), were recruited. Retention was 100% of the men and 98% of female partners. Attendance at group sessions was 63-73%. Between groups, mean (SD) weight changes among men were -3.4[.04] and -4.7[5.9] kg (p = 0.57) and among women -0.23[4.46] and -2.47[3.62] kg (p = 0.09), in the standard and enhanced groups. Conclusions: Weight losses from an intervention enhanced by partner involvement and an intervention with no partner involvement were not different. Treatment choice can be based on preference rather than outcome as both treatments are effective in producing clinically significant percent weight loss.Trial registration Clinical Trials NCT02458053.

Effectiveness of Diabetes Prevention Program translations among African Americans.

The Diabetes Prevention Program (DPP) demonstrated risk reduction for incident diabetes through weight loss among all participants, including African Americans. Several DPP translations have been conducted in less controlled settings, including primary care practices and communities; however, there is no detailed compilation of how effective these translations have been for African Americans. This systematic literature review evaluated DPP translations from 2003 to 2012. Eligible records were retrieved using a search strategy of relevant databases and gray literature. Retrieved records (n=1,272) were screened using a priori criteria, which resulted in 21 full-text studies for review. Seventeen studies were included in the full-text qualitative synthesis. Seven studies had 100% African American samples and 10 studies had mixed samples with African American subgroups. African American participants' average weight loss was roughly half of that achieved in the DPP intervention. However, with few higher-quality studies, small sample sizes and differences in intervention designs and implementation, comparisons across interventions were difficult. The suboptimal effectiveness of DPP translations among African American adults, particularly women, signals the need for enhancements to existing evidence-based interventions and more high-quality research that includes other at-risk African American subgroups such as men and younger adults of lower socioeconomic status.

The first pulse of the extremely bright GRB 130427A: a test lab for synchrotron shocks.

Gamma-ray burst (GRB) 130427A is one of the most energetic GRBs ever observed. The initial pulse up to 2.5 seconds is possibly the brightest well-isolated pulse observed to date. A fine time resolution spectral analysis shows power-law decays of the peak energy from the onset of the pulse, consistent with models of internal synchrotron shock pulses. However, a strongly correlated power-law behavior is observed between the luminosity and the spectral peak energy that is inconsistent with curvature effects arising in the relativistic outflow. It is difficult for any of the existing models to account for all of the observed spectral and temporal behaviors simultaneously.

Delayed developmental changes in neonatal vocalizations correlates with variations in ventral medial hypothalamus and central amygdala development in the rodent infant: effects of prenatal cocaine.

While variations in neonatal distress vocalizations have long been shown to reflect the integrity of nervous system development following a wide range of prenatal and perinatal insults, a paucity of research has explored the neurobiological basis of these variations. To address this, virgin Sprague-Dawley rats were bred and divided into three groups: [1] untreated, [2] chronic-cocaine treated (30 mg/kg/day, gestation days (GDs) 1-20); or [3] chronic saline treated (2 mg/kg/day, GDs 1-20). Pregnant dams were injected with Bromodeoxyuridine (10 mg/kg) on GDs 13-15 to label proliferating cells in limbic regions of interest. Ultrasonic vocalizations (USVs) were recorded on postnatal days (PNDs) 1, 14, and 21, from one male and female pup per litter. Variations in acoustic properties of USVs following cocaine-exposure were age and sex-dependent including measures of total number, total duration and amplitude of USVs, and percent of USVs with at least one harmonic. Following USV testing brains were stained with standard fluorescent immunohistochemistry protocols and examined for variations in neuronal development and if variations were associated with acoustic characteristics. Limbic region developmental differences following cocaine-exposure were sex- and age-dependent with variations in the ventral medial hypothalamus and central amygdala correlating with variations in vocalizations on PND 14 and 21. Results suggest maturation of the ventral medial hypothalamus and central amygdala may provide the basis for variations in the sound and production of USVs. As vocalizations may serve as a neurobehavioral marker for nervous system integrity, understanding the neurobiological basis of neonatal vocalizations may provide the basis for early intervention strategies in high-risk infant populations.

Incidence of rib fractures after stereotactic body radiotherapy for peripheral lung lesions: clinical experience and dose response estimation.

PURPOSE/OBJECTIVES: Late complications of SBRT include radiation related rib fractures. We estimate the incidence of rib fracturesas a function of maximum absorbed rib dose after stereotactic body radiotherapy (SBRT) for early stage lung cancer. MATERIALS/METHODS: Of 23 patients treated with image guided SBRT (60 Gy in 5 fractions) between 2003 and 2006, 4 developed pathological rib fractures near the SBRT planning target volume (PTV). Both planned maximum dose and maximum Fraction-size equivalent dose (FED) to the combined rib volume lying within the prescription isodose volume was determined and a probit dose response model was fitted to the observed rib fracture data for each. RESULTS: 17 patients were evaluated, all with a minimum of 15 months follow-up. Median followup was 43 months (range 15-60 months). The median time to rib fracture was 26.5 months (range 15-34 months). The maximum rib dose ranged from 23.8-74.7 Gy (median 57.8 Gy) in 5 fractions. Dose was a significant predictor of rib fracture (p=0.02), with a D50 (123FED50) estimate of 66.71 Gy (73.52 Gy). The steepness of the dose-response curve was quantified by the m and γ50 value, estimated at m = 0.1663 and γ50 = 2.39 for the maximum dose probit dose response model and at m = 0.2747 and γ50 = 1.45 for the maximum 123FED50 probit dose response model. CONCLUSIONS: Maximum rib dose should be carefully considered in SBRT with appropriate risk counseling of patients whose maximum rib dose exceeds a dose of 50 Gy in 5 fractions or a maximum 123FED50 of 43.1 Gy, estimated to be associated with a 6.6 % risk of rib fractures. Hence, the inclusion of ribs as an "organ at risk" in intensity modulated radiotherapy (IMRT) planning should be considered as a way to reduce the likelihood of rib fractures.

Initial clinical experience with stereotactic lung radiotherapy, based on a biological model-driven prescription method.

OBJECTIVES: A patient specific nomogram based biological dose selection (NBDS) model may allow for selection of a safe and effective dose schedule to treat early peripheral stage non-small cell lung cancer (NSCLC) with stereotactic body radiotherapy (SBRT). We report the initial clinical outcomes testing these concepts. METHODS: 23 patients with stage IA/B NSCLC were treated with SBRT. All patients had a prescription isodose volume/residual lung volume ratio of 2-3%, permitting a wide range of dose fractionation schemes under the NBDS-model that should yield a sufficiently low grade 2 or higher pneumonitis rate that the resultant long-term grade 3 or higher complication rate would be <20%. Based on the predications of the patient specific NBDS-model all patients could be safely treated using a modal prescription of 60 Gy in 5 fractions over 10 calendar days, with a median normalized tissue dose (NTD) of 122.4 Gy10. Kaplan-Meier analysis was performed to assess local control, overall, cause-specific and disease free survival. Toxicities and response rates were analyzed. RESULTS: Median follow-up was 43.2 months for all living patients. Analysis of 20 evaluable lesions demonstrated a major response rate of 80%. 3 year actuarial overall, cause-specific, and disease free survival, were 60, 79, and 55%, respectively. 3 year actuarial local control was 89%. Grade 2 or higher acute pulmonary toxicity was observed in 5 patients. The 1, 2 and 3-year actuarial incidence of grade 2 or higher pulmonary toxicity was 15, 27 and 27% (95% CI = 5 48%), with corresponding grade 3 incidence of 4, 10, and 10%. No grade 3 or higher non-pulmonary side-effects were observed. CONCLUSIONS: SBRT using a biological model-based fractionation scheme yields local control and survival rates comparable to other series that treat to higher NTDs; the pulmonary toxicity rate and grades are within the model-predicted parameters, but further follow-up is necessary for long-term validity of the model.

On the use of hyperpolarized helium MRI for conformal avoidance lung radiotherapy.

We wanted to illustrate the feasibility of using hyperpolarized helium magnetic resonance imaging (HPH-MRI) to obtain functional information that may assist in improving conformal avoidance of ventilating lung tissue during thoracic radiotherapy. HPH-MRI images were obtained from a volunteer patient and were first fused with a proton density-weighted (PD(w)) MRI to provide corresponding anatomic detail; they were then fused with the treatment planning computed tomography scan of a patient from our treatment planning database who possessed equivalent thoracic dimensions. An optimized treatment plan was then generated using the TomoTherapy treatment planning system, designating the HPH-enhancing regions as ventilation volume (VV). A dose-volume histogram compares the dosimetry of the lungs as a paired organ, the VV, and the lungs minus the VV. The clinical consequences of these changes was estimated using a bio-effect model, the parallel architecture model, or the local damage (f(dam)) model. Model parameters were chosen from published studies linking the incidence of grade 3+ pneumonitis, with the dose and volume irradiated. For two hypothetical treatment plans of 60 Gy in 30 fractions delivered to a right upper-lobe lung mass, one using and one ignoring the VV as an avoidance structure, the mean normalized total dose (NTD(mean)) values for the lung subvolumes were: lungs = 12.5 Gy3 vs. 13.52 Gy3, VV = 9.94 Gy3 vs. 13.95 Gy3, and lungs minus VV = 16.69 Gy3 vs. 19.16 Gy3. Using the f(dam) values generated from these plans, one would predict a reduction of the incidence of grade 3+ radiation pneumonitis from 12%-4% when compared with a conventionally optimized plan. The use of HPH-MRI to identify ventilated lung subvolumes is feasible and has the potential to be incorporated into conformal avoidance treatment planning paradigms. A prospective clinical study evaluating this imaging technique is being developed.

Clinical outcome analysis in "high-risk" versus "low-risk" patients eligible for national surgical adjuvant breast and bowel B-39/radiation therapy oncology group 0413 trial: five-year results.

PURPOSE: To report the local control and overall survival outcomes after lumpectomy followed by accelerated partial breast irradiation in high-risk patients as defined by the current inclusion criteria for the National Surgical Adjuvant Breast and Bowel B-39/Radiation Therapy Oncology Group 0413 Intergroup trial. METHODS AND MATERIALS: Between 2000 and 2005, 273 women with early-stage breast cancer were treated using either multicatheter interstitial brachytherapy (n=247) or MammoSite (n=26). Patients received 32-34 Gy in 8-10 twice-daily fractions using high-dose-rate 192Ir brachytherapy. All patients met the initial inclusion criteria for the Intergroup trial and were separated into two groups: high-risk patients (representing the cohort that remained eligible for the Intergroup trial) who satisfied one or more of the "high-risk" criteria (age<50 years, estrogen receptor negative, and/or positive lymph nodes; n=90), and low-risk patients who comprised the remainder of the cohort (n=183). The outcomes of the two cohorts were analyzed and compared. RESULTS: The median follow-up of the entire cohort was 48.5 months. No significant difference was found in outcomes at 5 years between the low- and high-risk groups, with a local control rate of 97.8% vs. 93.6%, crude local recurrence rate of 2.2% (n=4) vs. 4.4% (n=4), and overall survival rate of 92.1% vs. 89.5%, respectively. CONCLUSION: At 5 years, no statistically significant difference was found in outcomes for patients deemed to be at greater risk in the current National Surgical Adjuvant Breast and Bowel B-39/Radiation Therapy Oncology Group 0413 Intergroup trial. These clinical data support the inclusion of this "high-risk" population in this important ongoing study.

Photobleaching-based flow measurement in a commercial capillary electrophoresis chip instrument.

For microfluidic analytical instruments, a facile, fast, and accurate instrument test is highly demanded. The test includes the quantitative verification of the relationship between pressure drop and flow velocity for the hydrodynamic pump, between the electric voltage and electroosmotic flow (EOF) for the high-voltage supply, and the chip quality. The key point for the test is the measurement of the flow velocity. However, most currently available velocimetries cannot be directly used without any instrumental modification or adding extra instruments. We applied a recently developed Laser Induced Fluorescence Photobleaching Anemometer (LIFPA) for the instrument test through measuring fluid flow velocity in a microfluidic instrument with optical measurement without any modification and extra instrument. We have successfully used the method to test Caliper HTS 250 System from Caliper Life Sciences (Hopkinton, MA) with its own light source and detector. The experimental result demonstrates that this single-point method of measuring flow velocity can be easily used for accurate test of a microfluidic instrument in less than 10 min at extremely low cost without any modification and extra instrument.

Effects of mGlu1-receptor blockade on ethanol self-administration in inbred alcohol-preferring rats.

The Group I family of metabotropic glutamate receptors includes subtype 1 (mGlu1) and subtype 5 (mGlu5) receptors. This family of receptors has generated interest as potential targets for different areas of therapeutic development, including intervention for alcohol and drug abuse. Most of this interest is driven by findings showing involvement of mGlu5 receptors in the regulation of drug self-administration; however, studies examining the role of mGlu1 receptors in drug self-administration are limited. The purpose of this work was to examine the role of mGlu1-receptor antagonism in the maintenance of ethanol self-administration and the self-administration of an alternate nondrug reward, sucrose. Male alcohol-preferring inbred rats were trained to self-administer ethanol (15% vol/vol) versus water on a concurrent schedule of reinforcement, and the effect of the mGlu1-receptor antagonist JNJ16259685 (0.1-1.0mg/kg intraperitoneal [IP]) was evaluated on self-administration. The rats were then trained to self-administer sucrose (0.4% wt/vol) versus water, and the same dose range of JNJ16259685 was tested. Locomotor activity was tested in a separate assessment to evaluate potential nonspecific motor effects of the antagonist. Ethanol self-administration was dose dependently reduced by JNJ16259685. This reduction was likely due to a motor impairment as the lowest effective dose (0.1mg/kg) significantly reduced locomotor behavior. Sucrose self-administration was reduced by the highest JNJ16259685 dose (1.0mg/kg), and this reduction was also likely due to a motor impairment. Interestingly, ethanol self-administration was more sensitive to mGlu1-receptor antagonism than sucrose self-administration as lower JNJ16259685 doses reduced ethanol-reinforced responding and motor behavior. Together, these results suggest that mGlu1 receptors do not play a specific role in modulating ethanol self-administration or the self-administration of an alternate nondrug reward (i.e., sucrose).

Deletion of the 5-HT(3A)-receptor subunit blunts the induction of cocaine sensitization.

Serotonin (5-HT) receptors are classified into seven groups (5-HT(1-7)), comprising at least 14 structurally and pharmacologically distinct receptor subtypes. Pharmacological antagonism of ionotropic 5-HT(3) receptors has been shown to modulate both behavioral and neurochemical aspects of the induction of sensitization to cocaine. It is not known, however, if specific molecular subunits of the 5-HT(3) receptor influence the development of cocaine sensitization. To address this question, we studied the effects of acute and chronic intermittent cocaine administration in mice with a targeted deletion of the gene for the 5-HT(3A)-receptor subunit (5-HT(3A)-/-). 5-HT(3A) (-/-) mice showed blunted induction of cocaine-induced locomotor sensitization as compared with wild-type littermate controls. 5-HT(3A) (-/-) mice did not differ from wild-type littermate controls on measures of basal motor activity or response to acute cocaine treatment. Enhanced locomotor response to saline injection following cocaine sensitization was observed equally in 5-HT(3A) (-/-) and wild-type mice suggesting similar conditioned effects associated with chronic cocaine treatment. These data show a role for the 5-HT(3A)-receptor subunit in the induction of behavioral sensitization to cocaine and suggest that the 5-HT(3A) molecular subunit modulates neurobehavioral adaptations to cocaine, which may underlie aspects of addiction.

Are we influencing outcome in oropharynx cancer with intensity-modulated radiotherapy? An inter-era comparison.

PURPOSE: To analyze the outcome in all oropharynx cancer patients treated at the University of Wisconsin during 1995-2005 and highlight the methodologic challenge in comparing outcome after intensity-modulated radiotherapy (IMRT) with that of historical controls. METHODS AND MATERIALS: Outcomes were compared in 195 oropharynx cancer patients after definitive radiotherapy with curative intent in the pre-IMRT era (pre-IMRT, n = 105), after IMRT (IMRT+, n = 52) or after non-IMRT techniques during the IMRT era (IMRT-, n = 38). RESULTS: With a median follow-up of 30.4 months, the 3-year overall survival rate in IMRT+, IMRT-, and pre-IMRT patients was 88.2%, 81.1%, and 67.7%, respectively; and for locoregional control was 96.1%, 78.1%, and 81.1%. Patients from the IMRT era more frequently received concurrent chemotherapy (67% vs. 6%, p < 0.001) and underwent adjuvant neck dissection (52% vs. 29%, p = 0.002). Patients with T3-4 disease and bilateral neck disease were significantly less likely to receive IMRT. Cox regression analysis identified IMRT as a significant prognostic factor (p = 0.04); however, after including T stage in the model, IMRT lost independent significance (p = 0.2). Analysis of a potential effect of IMRT on Grade 3+ mucositis or skin reaction was also hampered by the change in other treatment characteristics. CONCLUSIONS: Outcomes in oropharynx cancer have improved at our institution since the introduction of IMRT. However, multiple factors have contributed to this improvement, and presentation of IMRT outcomes without the full context of historical and contemporary controls may yield data that overstate outcome after IMRT.

Increased response to morphine in mice lacking protein kinase C epsilon.

The protein kinase C (PKC) family of serine-threonine kinases has been implicated in behavioral responses to opiates, but little is known about the individual PKC isozymes involved. Here, we show that mice lacking PKCepsilon have increased sensitivity to the rewarding effects of morphine, revealed as the expression of place preference and intravenous self-administration at very low doses of morphine that do not evoke place preference or self-administration in wild-type mice. The PKCepsilon null mice also show prolonged maintenance of morphine place preference in response to repeated testing when compared with wild-type mice. The supraspinal analgesic effects of morphine are enhanced in PKCepsilon null mice, and the development of tolerance to the spinal analgesic effects of morphine is delayed. The density of mu-opioid receptors and their coupling to G-proteins are normal. These studies identify PKCepsilon as a key regulator of opiate sensitivity in mice.

A chemogenomic approach to discovering target-selective drugs.

An approach is described for identifying scaffolds with predefined selectivity profiles directly from a high-quality high-throughput screening data set, and monitoring that selectivity profile throughout the drug discovery process. This approach results in reduced drug development timelines from "hit" to "lead" stage, and should reduce liabilities for a molecule advancing through the development process.

Anomalous functional organization of barrel cortex in GAP-43 deficient mice.

Growth associated protein 43 (GAP-43), found only in the nervous system, regulates the response of neurons to axon guidance signals. It is also critical for establishing normal somatotopy. Mice lacking GAP-43 (KO) show aberrant pathfinding by thalamocortical afferents, and do not form cortical whisker/barrels. GAP-43 heterozygous (HZ) mice show more subtle deficits--delayed barrel segregation and enlarged barrels at postnatal day 7. Here, we used cortical intrinsic signal imaging to characterize adult somatotopy in wildtype (WT), GAP-43 KO, and HZ mice. We found clear foci of activation in GAP-43 KO cortex in response to single-whisker stimulation. However, the KO spatial activation patterns showed severe anomalies, indicating a loss of functional somatotopy. In some cases, multiple foci were activated by single whiskers, while in other cases, the same cortical zone was activated by several whiskers. The results are consistent with our previous findings of aberrant pathfinding and clustering by thalamocortical afferent axons, and absence of barrel patterning. Our findings indicate that cortex acts to cluster afferents from a given whisker, even in the absence of normal topography. By contrast, single-whisker stimulation revealed normal adult topographic organization in WT and HZ mice. However, we found that functional representations of adult HZ barrels are larger than those found in WT mice. Since histological HZ barrels recover normal dimensions by postnatal day 26, the altered circuit function in GAP-43 HZ cortex could be a secondary consequence of the rescue of barrel dimensions.

Use-dependent plasticity in barrel cortex: intrinsic signal imaging reveals functional expansion of spared whisker representation into adjacent deprived columns.

We used optical imaging of intrinsic cortical signals, elicited by whisker stimulation, to define areas of activation in primary sensory cortex of normal hamsters and hamsters subjected to neonatal follicle ablation at postnatal day seven (P7). Follicle ablations were unilateral, and spared either C-row whiskers or the second whisker arc. This study was done to determine if the intrinsic cortical connectivity pattern of the barrel cortex, established during the critical period, affects the process of representational plasticity that follows whisker follicle ablation. Additionally, we tested the ability to monitor such changes in individual cortical whisker representations using intrinsic signal imaging. Stimulation of a single whisker yielded peak activation of a barrel-sized patch in the somatotopically appropriate location in normal cortex. In both row and arc-spared animals, functional representations corresponding to spared follicles were significantly stronger and more oblong than normal. The pattern of activation differed in the row-sparing and arc-sparing groups, in that the expansion was preferentially into deprived, not spared areas. Single whisker stimulation in row-spared cases preferentially activated the corresponding barrel arc, while stimulation of one whisker in arc-spared cases produced elongated activation down the barrel row. Since whisker deflection normally has a net inhibitory effect on neighboring barrels, our data suggest that intracortical inhibition fails to develop normally in deprived cortical columns. Because thalamocortical projections are not affected by follicle ablation after P7, we suggest that the effects we observed are largely cortical, not thalamocortical.