Alternatively, splenocytes were cultured

in the presence or absence of peptides VNHRFTLV or TsKb-20 and the expression of surface CD107a, http://www.selleckchem.com/products/PF-2341066.html IFN-γ and TNF-α by ICS. In infected mice, administration of FTY720 resulted in 2.52- or 3.05-fold increases in the frequency of IFN-γ-secreting cells from the LNs specific for VNHRFTLV or TsKb-20, respectively, as detected using the ELISPOT assay (Fig. 6). In contrast, this increase in the frequency IFN-γ-secreting peptide-specific cells in the LN was accompanied by a significant decrease of immune responses of splenic lymphocytes. Immune responses were initially determined by the frequency of IFN-γ-producing cells as measured by the ELISPOT assay (Fig. 7A). The frequency of IFN-γ-producing cells found in the spleen after FTY720 administration was reduced by 74.55% or 100% upon stimulation with peptides VNHRFTLV or TsKb-20, respectively (Fig. 7A). Subsequently, we estimated the immune response by the detection of peptide-specific CD8+ cells that mobilized CD107a to their surface and expressed IFN-γ and TNF-α upon exposure to the peptides in vitro. The frequency of CD8+ cells that were CD107a+, IFN-γ+

or TNF-α+ was reduced by 74.61% or 84.15% after stimulation Reverse Transcriptase inhibitor with VNHRFTLV or TsKb-20, respectively ( Fig. 7B). The reduction substantially affected all the different subpopulations of CD8+ cells ( Fig. 7C). The proportions of each population did not change significantly in the cells collected from infected mice that were administered or not with FTY720 ( Fig. 7D). To evaluate the influence of restricting T-cell re-circulation on the outcome of infection, we also monitored the parasitemia levels and survival of

mice that were and were not subjected to FTY720 over the course of infection. We found that drug exposure resulted in increased parasitemia and accelerated mortality of Morin Hydrate infected mice (Fig. 8A and B, respectively). Therefore, we concluded that lymphocyte re-circulation is indeed important for the acquired protective immune response in this mouse model of acute infection. We then sought to test the same hypothesis by applying a distinctly different approach. In this case, we used highly susceptible A/Sn mice that were genetically vaccinated by priming with plasmid pIgSPCl.9 followed by a booster immunization with AdASP-2. We previously showed that this heterologous prime-boost regimen reproducibly conferred protective immunity against a lethal challenge with T. cruzi [25]. Immunity was mediated by CD8+ T cells as depletion of these T cells renders these mice completely susceptible to infection. These CD8+ T cells are specific for the ASP-2 H-Kk restricted epitopes TEWETGQI, PETLGHEI or YEIVAGYI [31]. Prior to challenge, these mice exhibit a strong immune response to all three epitopes [31]. Following infection (s.c.), some of these vaccinated mice were subjected to FTY720. We then monitored the parasitemia levels and survival.

Seven important factors have convinced authorities to prioritise prevention: declining life expectancy, rising disease risk, impending cost burden, broad social impact, inequity of risk, cost effectiveness, and efficacy. 1. The life expectancy at birth of Australians is very good (84 years for females, 79 years for males), ranking third internationally (AIHW 2010). Life expectancy in Australia this website rose from 59/55 years early in

the twentieth century to 70/65 years by mid-century due to better management of infectious disease and better hygiene and living standards. However, mid-century life expectancy plateaued and actually declined for males due to chronic lifestyle diseases especially cardiovascular disease. Improved tertiary management of chronic disease has continued the increase in life expectancy since then. But once again there is downward pressure on life expectancy, with estimates

that the impact of obesity alone is equivalent to a 2-year decline in life expectancy at a population level (D’Arcy and Smith, 2008). Tobacco smoking, alcohol consumption, low fruit and vegetable intake, high body mass, and physical inactivity account for an estimated 27% of the total Australian health burden (Begg et al 2007) through pathways to cancer, chronic obstructive pulmonary disease, heart disease, stroke, accidents, suicide, diabetes, and Wortmannin solubility dmso other disorders (AIHW 2010). Further, these risk behaviours often cluster

together (NPHP 2001). 1. Tobacco is smoked by only about 19% of Australian adults now Anacetrapib (AIHW 2010), but this and the legacy of prior higher rates means it accounts for ~8% of the total health burden in Australia (Begg et al 2007). The preventive guideline is to avoid smoking. Despite advances in tertiary care, the health of populations in affluent countries is declining. The impending cost burden of dealing with lifestyle-related health disorders will overwhelm current health service delivery models. Therefore we must prioritise prevention now to optimise the health of the population. Currently there is a window of opportunity created by government urgency to reform health systems and support other preventive initiatives to reduce the impending disease burden. Physiotherapists could play a major role in preventive health – but if we don’t there are many other groups who will take on this vital role for our society. A desire to help people live healthier, happier, and more functional lives by reducing the burden of disease and injury is a driving motivation to enter the physiotherapy profession and to remain a physiotherapist. As a profession we have long promoted the notion to ‘move well, stay well’.

Accordingly, empirical studies investigating emotion regulation have grown exponentially over the last two decades,

reflecting mounting interest within the field (Gross, 2013). Despite the broad scientific interest in understanding how emotions are regulated, however, the notion that stress may be detrimental to emotional control has been relatively overlooked within this literature. Consequently, the effects of stress on the capacity to flexibly control emotional responses have remained largely unexplored. The studies reviewed here offer some initial insight into understanding how acute stress exposure affects the inhibition and control of conditioned fear. The research discussed in this review used Pavlovian fear conditioning as selleck inhibitor a basis for understanding the effects of stress on the regulation of fear. Since the neural circuitry underlying fear learning is highly

conserved across species, we can use CP-673451 chemical structure animal models as a basis for understanding how stress may influence this circuitry in humans as well. Our investigation of extinction and cognitive regulation reveals robust effects of stress impairing the persistent inhibition of fear, presumably by altering prefrontal cortex function. Although less is known concerning the impact of stress on the persistent fear reduction observed with avoidance and reconsolidation, it is possible these fear regulation techniques are less vulnerable to the negative consequences of stress since they rely less on the inhibitory mechanisms involved in extinction and cognitive regulation. It is important to note that the behavioral and neural research covered in this review focused mainly on brief exposure to stress, rather

than chronic exposure. Although the immediate effects of acute stress can exert detrimental effects on the brain regions critical to the regulation of fear responses, chronic exposure to stress can trigger to more systemic neuroendocrine changes. For example, chronic stress can lead to dysfunctional regulation of the HPA-axis, resulting in a flattened diurnal cycle of cortisol release, such as that seen in depressives and PTSD (Young et al., 1994; Yehuda, 2009). It can also lead to more profound structural Dynein and functional changes in brain regions critical to autonomic and HPA-axis related regulation (i.e., amygdala and hippocampus) that can lead to suppression of synaptic plasticity and neurogenesis in these regions (see McEwen, 2003 for review). Collectively, chronic stress produces what has referred to as allostatic load, creating an overwhelming demand on the neural circuits that mediate appropriate responses and recovery from stress. Fear learning and regulation is a prominent model for describing the pathogenesis of anxiety disorders and stress-related psychopathology.

Different strategies have been applied for linking antibodies with DNA templates, like streptavidin bridge combined with biotinylated antibody and biotinylated DNA template, or chemically conjugated antibody-DNA complexes ( Lind and Kubista,

2005 and Niemeyer et al., 2007). The amount of DNA amplified during PCR corresponds to the amount of target structure recognized by the antibody, ubiquitin-Proteasome degradation and can be detected by electrophoresis ( Zhou et al., 1993) or by ELISA, utilizing digoxigenin- or biotin-labeled PCR products ( Niemeyer et al., 1997 and Smrž and Dráber, 2003). Later, immunodetection was combined with real-time PCR and used for quantification of vascular endothelial grow factor ( Sims et al., 2000). The method was further modified in such a way that both protein detection

and real-time PCR were performed in the same well of the TopYield strip ( Niemeyer et al., 2007). Furthermore, a gold nanoparticle (Au-NP)-based bio-bar code assay for ultrasensitive detection of proteins has been developed. The assay utilizes Au-NPs functionalized with both thiolated single-strand DNA oligonucleotide and an antibody to the target antigen ( Nam et al., 2003, Nam et al., 2004 and Georganopoulou et al., 2005). Finally, PCR assays based on antibody- and oligonucleotide-functionalized Au-NPs were used for detection of Hantaan virus nucleocapsid protein ( Chen et al., 2009) and respiratory syncytial viruses ( Perez et al., 2011). Although the assays showed high sensitivity for virus detection, they required two sets of wells (for immunodetection and PCR) and therefore were not suitable for high throughput screening and were fraught with high risk of contamination. Here MG-132 research buy we tested

the suitability of functionalized Au-NPs-based iPCR (Nano-iPCR) for detection of low concentrations of cytokines in cell culture supernatants, and changes in cytokine concentration in aging cultures of BMMCs. We defined the conditions for simplified detection of cytokines by Nano-iPCR, and compared the performance of assays based on antibodies anchored either directly on the plastic surface or through extravidin. The assays were carried out in PCR polypropylene wells or wells of TopYield polycarbonate strips which allow more efficient binding of antibodies. We further compared Nano-iPCR with iPCR and PFKL ELISA; outline of the assays is shown in Fig. 1. For these comparisons we utilized identical immunoreagents in all assays. The data indicate that Nano-iPCR offers a sensitive, rapid and robust assay for detection of low concentrations of cytokines in complex biological fluids. Advantages and drawbacks of different assays are discussed. Rabbit anti-murine IL-3 and rabbit anti-murine SCF polyclonal antibodies and their biotinylated forms, recombinant (r) murine IL-3 and rSCF were all obtained from PeproTech (London, UK). Colloidal Au-NPs (30 nm), containing approximately 2 × 1011 Au-NPs/ml, were obtained from BBInternational (Cardiff, UK).

We conducted western blot analysis to examine the protein level of ASK1 (Fig. 2A) and VEGF (Fig. 2B), which is known to play important roles in vascular permeability following OGD/R. This data shows the protein level in various reperfusion time points (reperfusion 0 min, 30 min, 1 h, and 3 h) after OGD (Fig. 2). VEGF protein expression was significantly increased

at reperfusion 0 min after OGD. VEGF protein level was augmented from reperfusion 0 min Gefitinib to 30 min. However, they were gradually decreased from reperfusion 1–3 h after OGD (Fig. 2A). Western blotting was also performed to evaluate ASK1 expression in OGD/R injured bEND.3.cells (Fig. 2B). The protein level of ASK1 was highly augmented after hypoxia injury and especially peaked at reperfusion 30 min after OGD. ASK1 protein level was gradually decreased in bEND.3.cells from reperfusion 1–3 h after OGD. This result suggests that ASK1 may be associated with the expression of VEGF in brain endothelial cells after cerebral ischemia. Also, ASK1 and VEGF may activate at the similar Cyclopamine in vivo time point after cerebral

ischemia. To examine whether ASK1 directly affects the expression of VEGF in brain endothelial cells during OGD/R injury, we treated ASK1 inhibitor (NQDI-1) in bEND.3.cells before OGD/R injury. Fig. 3 shows that inhibition of ASK1 activity using NQDI-1 reduced the protein level of phosphorylation-ASK1 and VEGF compared to the OGD/R group at reperfusion 30 min after hypoxia injury (Fig. 3A and B). Our data suggest that ASK1 might play an important role in VEGF expression in brain endothelial cells after hypoxic injury. Furthermore, ASK1 may modulate the expression of VEGF at reperfusion early time point after OGD. To investigate whether ASK1 inhibition affects vascular permeability in

animal brain, we measured brain edema at reperfusion 24 h after MCAO injury using TTC staining (Fig. 4A). White areas in brain are damaged brain areas due to ischemia (Fig. 4A). The graph shows the percentage of the ipsilateral hemisphere compared with the contralateral hemisphere both in the MCAO and si-ASK+MCAO groups (Fig. 4B). The percentage of brain edema in the MCAO group was >20% whereas the percentage of brain edema after si-ASK1 treatment was <10%. Brain edema (%) was significantly DOK2 reduced in the si-ASK1+MCAO group compared with the MCAO group. Our results indicate that the inhibition of ASK1 reduced brain edema formation after ischemic brain injury. Considering this finding, the inhibition of ASK1 may be a useful strategy for reducing brain edema. Cresyl violet staining was performed at reperfusion 24 h after MCAO injury to histologically assess the extent of ischemia-induced damage in the striatum and cortex (Fig. 5). In the NON group (without MCAO injury, without ASK1-siRNA treatment), intact cellular structure was observed in both the cortex and striatum.

, 2010a). Making better choices concerning food acquisition, based on individual knowledge about food and healthiness, continues to be a challenge, due to the great diversity of food products available nowadays. It is essential to emphasize the importance of updating specific food legislation, once this is a highly changeable industry and consumers are increasingly

demanding ROCK inhibitor for newness. Also, a more uniform legislation would certainly contribute for globalization. In the present study, the improvement of the guava mousses’ nutritional values was possible, particularly regarding the fat content, once the vast majority of modified mousses had a considerable reduction in this nutrient content through the substitution of fat milk for inulin and/or whey protein concentrate. Also, the addition of inulin and FOS in these mousses was decisive for the contribution regarding dietary fibre. Based on the results of this and the previous studies of this research group with guava mousses, MF–I–WPC selleck compound was the formulation that fit the most of desirable features: improvement of energy, total and saturated fat, protein and dietary fibre content, good viability of L. acidophilus during storage conditions (refrigeration and freezing) and survival of this microorganism in the simulated gastrointestinal fluids, besides presenting texture and sensorial acceptability comparable to control mousse

4-Aminobutyrate aminotransferase MF. The authors wish to thank to Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (Projects 06/51297-0, 05/51317-8, 04/13597-6, 04/05972-1, 08/55061-6, and 09/07160-8), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Conselho Nacional de Desenvolvimento

Científico e Tecnológico (CNPq) for financial support and scholarships; and Christian Hansen, Clariant, Danisco, Kienast & Kratschmer, Orafti and Purac Sínteses companies for providing part of the material resources employed in this study. The authors gratefully acknowledge Alexandre Mariani Rodrigues for his technical assistance and Alexandra Tavares de Melo for her useful advice and valuable comments on food legislation and claims. “
“Free radicals, reactive oxygen species (ROS) and reactive nitrogen species (RNS), are constantly produced by cells during normal and pathological energy metabolism. Both ROS and RNS have been associated with many diseases and degenerative processes in aging (Halliwell, 2000). Almost all organisms are well protected against free radical damage by antioxidant enzymes such as superoxide dismutase and catalase. However, these systems are frequently insufficient to totally prevent the damage, resulting in diseases and accelerated aging. Natural products obtained through the diet, such as tocopherol, ascorbic acid, carotenoids and phenolic compounds with antioxidant activity can be useful to reduce oxidative damage in the human body.

Greenberger Thomas Gremmel Weihua Guan Prajwal Gurung Kirk Hamilton Joshua Hare David Harris Daniel Hayes Chuan He Steffen Heeg Britney Helling Norah Henry Eli Hershkovitz Helen Heslop Jeffrey Hodgin Mimi Hu R. Stephanie Huang H.David Humes Warrick Inder Allan Jaffe Manu Jain Edward N. Janoff Craig Jefferies Sonata Jodele Duncan Johnstone

Michelle Kahlenberg Ravi Kalhan Nigel S. Key Farrah Kheradmand Seong-Kyu Kim Michael King Petra Kleinbongard Hon Wai Koon Sean Koppe Krishnan Koyamangalath Lucia Kucerova Yoshiki Kusama Richard Lafayette Luigi Laghi James Lane Irene Lang Benjamin Laskin Rodrigo Leal Andrew Leask Emilia Lecuona Joshua Leonard Kevin Leslie Edward Lesnefsky Maciej Lesniak Paul ATM/ATR inhibitor Lewis Yi Li ES Lianidou Weei-Chin Lin Shing-Jong Lin De Lin Marc Lippman Wei Liu Sumei Liu Gang Liu Fei Liu Dakai Liu Emil Lou Alessandro Lugli Malcom MK-2206 manufacturer Macleod Meena Madhur Lars Maegdefessel Patrudu Makena Deepak Malhotra Sunil Mallanna Massimo Mangino A.J. Marian Cary Mariash Philipp Mario Caroline Marshall George Martin James Martins Philip Mason Biji Mathew Sandra McAllister Kim McBride Susanna McColley Akira

Meguro Farrell Mendelsohn Steven Mentzer Jordan Metcalf Martha Mims

SALVATORE MINISOLA Abhisek Mitra Nicholas Mitsiades Markus Mohaupt Aaron Mohs Zahra Montazeri Daniel Musher Roland Nau Georges Nemer Paul Ney Dennis E. Niewoehner Timothy B Niewold Shuji Ogino Jill Ohar Gil Omenn Giuseppe Orlando Carl Orringer Tadeusz Osadnik John O’Toole Gavin Oudit Ratnasari Padang Vasantha Padmanabhan Udai Pandey Francisco Pan-Montojo Ralph J. Panos Choul Yong Park Linda Partridge Subramaniam Pennathur Maikel Peppelenbosch Maria Pereira Francisco Campos Pérez Eileen Pernot Phillip K. Peterson Richard Phipps Massimo Pietropaolo Irina Pinchuk Edoxaban Graham Poage Catherine Poh Michael Polymenis Bogdan Popescu Kailash Prasad Josef Prchal Vasu Punj Edward Purdue Hershel Raff Nalini Rajamannan Narayan Ramakrishna Nithya Ramnath Toralf Reimer Jun Ren Robert Roberts Leonardo Roever Sharon Rosenberg Myrna Rosenfeld Ann Rosenthal Catharine Ross Charles Rosser David Roth Anita Sabichi Joshua Safer Hiroshi Saito Nathan Sandbo Paul Sanders Robert Sargis Akinori Sato Amr Sawalha Amnon Schlegel Paul Schmidt Bryan Schneider Andreas Schwingshackl Sudhir V.

MB9), the a*ph(λ) spectrum of the surface water was flat in the blue-green region. The surface Chl a concentrations on the NS transect were generally lower (< 5 μg l− 1) and a*ph(λ) values were high (≥ 0.003 m2(mg TChl a)− 1) at most of the stations ( Figure 7). At the stations where the marker pigment for diatoms was high (> 0.5 μg l− 1), very low ā*ph(λ) values (≥ 0.003 m2(mg TChl a)− 1) were recorded. The blue-red peak ratios (a*ph(440) : a*ph(675)), a reflection of accessory pigment absorption as well as of pigment packaging, showed a value of 1.45 ± 0.56 (mean ± SD)

on the NS transect and 1.56 ± 0.38 on the EW transect. Comparatively low ratios (< 1.2) of a*ph(440) : a*ph(675) at stn. MB12 could be associated with the combined effect of packaging and relatively elevated ratios of accessory pigments like fucoxanthin, peridinin and diadinoxanthin. click here The marked absorption peaks at 455 nm and 653 nm at almost all stations can also be attributed to a high ratio of Chl b to TChl a. It was observed that the fourth derivative of the absorption spectra was useful for identifying pigment peaks (Figure 8). At most stations Chl a absorption maxima were found around wavelengths 440 and 675 nm, while accessory pigments displayed their absorption peaks in the 490–550 nm regions.

The stations on the north-south transect showed small peaks in the 560–618 nm region, which could be accounted BMS-354825 in vitro for by degradation products and Chl c. Fucoxanthin peaks could be identified in the 521–530 nm region in the stations towards the northern

part of the bay. The diadinoxanthin peak was detectable at 425–500 nm at most stations. Phycoerythrobilin was suggested check details for the peak at 548 nm. Smaller peaks were observed in the 589–594 nm region and also at 627 and 647 nm. The regression of the chlorophyll absorption maxima at the red region with the chlorophyll a concentration showed a good correlation for chlorophyll a (r2 = 0.71, n = 39). The a*ph values recorded in the present study are typical of eutrophic waters, and such values are similar for a diatom dominated condition ( Prézelin & Boczar 1986). The inverse correlation observed in this study between chlorophyll-specific absorption and Chl a concentration (y = − 291.65 + 8.5873; r2 = 0.268, n = 29) is well documented in many previous studies ( Prieur and Sathyendranath, 1981, Bricaud et al., 1995, Bricaud et al., 2004, Cleveland, 1995, Ciotti et al., 1999 and Sathyendranath et al., 2001). There was a pronounced variation in the values of a*ph(440) at the centre of the bloom patch and beyond it. At stn. MB9, where the highest chlorophyll concentration was observed at the surface, the value of a*ph(λ) was very low, varying by two orders of magnitude with respect to the nearby stations (stn. MB7, MB8 and MB13) (see Figure 7).

Therefore, the structure of cellulosic biomass must be pretreated prior to enzymatic hydrolysis to make cellulose more accessible to enzymatic Selleckchem Bortezomib conversion [29] and [11]. Various physical, chemical, physico-chemical and biological pretreatment methods have been well-investigated for ethanol production from lignocellulosic biomass [36], [16] and [35].

The purpose of the pretreatment is mainly to increase the accessibility of the enzymes to cellulose the by solubilisation of hemicelluloses or/and lignin, and by decreasing the degree of polymerization and cellulose fibre crystallinity [12]. Moreover, adding surfactants has also improved the effectiveness of the cellulose hydrolysis [3] and [10]. To improve the rate of enzymatic hydrolysis, researchers have focused on the study of multiple enzymatic hydrolysis process parameters, including substrate concentration, and reaction conditions such as hydrolysis time, pH, temperature and addition

of surfactants [35]. Optimal parameters are highly dependant on the physico-chemical structure of the digested biomass, and different pretreatment methods will produce substantially different biomass. Pretreatment in a twin-screw extruder can be used (among other things) to hydrolyze and remove the hemicellulose fraction [23], [24] and [7]. However, the effect of xylose removal via extrusion pretreatment, EGFR inhibitor along with other process parameters on the enzymatic hydrolysis of corncobs, has not yet been systematically characterized. In the present study, two differently extruded corncobs with 7% xylose removal and 80% xylose removal, GPX6 respectively, were used as a source of enzymatic hydrolysis. The characteristics of these two materials were examined by SEM and XRD. A face-centered

central composite design was used to study the combined effects of various enzymatic hydrolysis process variables (enzyme loading, surfactant addition, and hydrolysis time) with these two extruded corncobs (7% xylose removal, 80% xylose removal). Corncobs were obtained from local farmers in Chatham, ON, Canada. Corncobs were cleaned and ground to the particle size of 0.5–1 cm3 and moisture was adjusted to 50% dry matter. Corncobs were then fed into a continuous steam explosion pretreatment reactor (GreenField Ethanol, Chatham). The reactor was set at a temperature of 205 °C with pH 4.8 in a system pressurized with saturated steam. The overall retention time of the corncobs during pretreatment was 5 min. Hemicellulose was hydrolyzed to xylose or xylo-oligosaccharides under these conditions. The pressure of the reactor was rapidly released to atmospheric pressure, thus the pressurized corncobs were flashed into a cyclone separator, which increased the accessible surface area of the fibres for the enzymes.

However, this does not necessarily mean that all of these cells belong to the spinoparabrachial tract, since some of the labelling may result IDO inhibitor from uptake of tracer by fibres passing through the injection site. For example, the projection from lamina I to the PAG passes through the rostral part of the parabrachial area (Bernard et al., 1995 and Feil and Herbert, 1995), and although there is a dense terminal arborisation within the LPb it is possible that some axons pass through this region without contributing to this arborisation. If this is the case, then some spino-PAG neurons would not belong to the spinoparabrachial tract, but may be retrogradely labelled

from the LPb. Spinothalamic axons from lamina I ascend near the parabrachial area and are located approximately 500 μm lateral to the external lateral nucleus of the LPb (J.F. Bernard, personal communication). Although these axons are likely to have been included in the LPb injections in several of the

present series of experiments, this should not alter the interpretation, because our previous finding that virtually all spinothalamic lamina I neurons were labelled from LPb was obtained from cases in which the LPb injections did not extend into this region (Al-Khater and Todd, 2009). The uptake of tracer by fibres of passage is unlikely learn more to have contributed to the labelling from the dorsal medulla, as these injections Demeclocycline were located a considerable distance from the main ascending bundle of axons from lamina I, which is in the ventrolateral part of the brainstem at this level (Mehler, 1969, Zemlan

et al., 1978 and Slugg and Light, 1994). However, it causes a significant problem for interpreting the labelling that results from injections of tracer into the CVLM, as we have reported previously (Spike et al., 2003). Although tracer injections into CVLM cannot be used to identify supraspinal targets, they are useful because they can label a very high proportion of lamina I projection neurons in both enlargements. Our previous estimate that there were ∼ 400 lamina I projection neurons on each side in the L4 segment of the rat was based on counts of cells retrogradely labelled from LPb, CVLM and PAG (Spike et al., 2003), and we have since demonstrated that all spinothalamic lamina I cells at this level are included in the population labelled from LPb (see above). Since nearly all lamina I neurons that project to the dorsal medulla are also labelled from LPb, this provides further support for the reliability of our estimate. The present results, together with those of Al-Khater and Todd (2009) suggest that virtually all lamina I projection neurons in C7 can also be labelled from LPb.