One study also assessed the effect of viral load (VL) on sperm parameters and found a negative correlation with sperm motility and morphology [14]. Our early analysis again suggested a more consistent effect, with a significant

positive correlation observed between CD4 cell count and sperm concentration, total count, progressive motility and post-preparation concentration and a significant negative correlation with normal sperm morphology of both raw and post-preparation samples. At the numbers then available, no correlation was observed between VL, years since diagnosis, use of antiretrovirals or duration of antiretroviral use and any sperm parameter [18]. The aim of the present study was to present a decade of data from the SWP programme in the UK to demonstrate the effect of markers of HIV disease progression and treatment on seminal parameters. The pretreatment Pexidartinib manufacturer work-up SRT1720 chemical structure has been discussed fully elsewhere [19]. In brief, a full fertility and sexual health screen is performed

on both partners to define the optimum treatment modality, exclude HIV coinfection and treat any genital lesions or infections that may increase the risk of viral transmission [20]. Our recommendations are that all patients should receive careful preconceptual counselling, both together and individually, before embarking on treatment [21], where the nature and risks of sperm washing, the impact of possible treatment failure, the issues involved in coping with a child when one parent is

HIV positive, and the possibility of having to cope as a single parent are discussed. In particular, it is mandatory that both partners understand sperm washing to be a risk-reduction method and not a risk-free method as, technically, the virus could still be present in the washed sample at a titre below the detection limit of the HIV assay. Although there have been no reports of seroconversion in the female partner when semen has been correctly processed in the 3315 cycles published thus far by the Centre for Reproductive PAK6 Assisted Techniques for HIV in Europe (CREAThE) network [22], the possibility of viral infection of the woman and subsequent child still exists, and the alternative risk-free option of donor insemination should be discussed and appropriate consent obtained from both partners, including confirmation of this information. Raw and post-preparation semen parameters from 439 samples used for cycles of IUI were correlated with markers of HIV disease (CD4 cell count and VL), use of HAART, duration of disease and duration of HAART. HIV history was confirmed using a questionnaire at the initial visit and the most recent CD4 cell count and VL, as well as the medication history, were confirmed at the time of the production of a sample for treatment.

The membranes were counterstained using corresponding donkey anti-guinea pig (1 : 5000; Jackson Immunoresearch, West Grove, PA, USA), goat anti-rabbit or anti-mouse (both 1 : 3000; Bio-Rad Laboratories, Hercules, CA, USA) horseradish peroxidase conjugates. For stripping between the immunoblot procedures, membranes were rinsed and incubated in Restore Western Blot Stripping Buffer (Thermo Scientific, Rockford, IL, USA) according to the manufacturer’s instructions. For visualization of the proteins, the membranes were exposed to the enhanced chemiluminescence detection system Lumigen PS-3 (1 : 40; GE Healthcare, Buckinghamshire, UK). No immunopositive bands were observed

when immunoblotting was performed with anti-CB1 antibodies pre-absorbed with the antigene peptide (5 μg/mL; Frontier Science, Japan). For immunoprecipitation, ~2.0 mg of total protein from mouse embryo (E16.5) brain mitochondrial fractions

(prepared Selleck Dasatinib as above) was incubated overnight at +4 °C with 3 μL of made-in-guinea pig anti-CB1 sera (Frontier Science, Japan). Thirty microliters of a 1 : 1 slurry of protein A-sepharose (GE Healthcare, Buckinghamshire, UK) in phosphate-buffered saline was then added and antibody-bound protein was collected during a 2-h incubation at +4 °C. Seliciclib The Sepharose beads were washed four times in 500 μL phosphate-buffered saline containing protease inhibitor cocktail (1 : 500; Calbiochem, La Jolla, CA, USA). The beads and bound protein were loaded in mini gel and separated using electrophoresis as above. The gel was then stained with SimplyBlue colloidal Coomassie (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s instructions. The ~40-kDa band was cut from the gel and destained

in three washes of acetic acid : methanol : H2O (10 : 50 : 40) solution. The sample was submitted for in-gel tryptic digestion, followed by liquid chromatography, quadrupole/time-of-flight tandem mass spectrometry and peptide mass database searching (Keck Facility, Yale University, New Haven, CT, USA). Mouse neuroblastoma 2A cells were cultured in Dulbecco’s D-MEM/F12 medium containing 9% fetal bovine serum (all from Sigma-Aldrich, St Louis, MO, USA). For transfections, we cloned full-length SLP-2 from E14.5 embryo brain cDNA into pIRES2-EGFP (Clontech, Mountain View, CA, USA); transfections with pEGFP Thymidylate synthase (Clontech, Mountain View, CA, USA) were used as negative controls. Newly passaged cells at about 70–80% confluency were starved of serum overnight and transfected with 5 μg SLP-2 DNA using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s guidelines. After 24 h, cells were washed in phosphate-buffered saline, and immediately scraped and lysed in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) containing protease (Roche, Indianapolis, IN, USA) and phosphatase (Sigma-Aldrich, St Louis, MO, USA) inhibitor cocktails.

Assuming that bodies returned for cremation represented 57.4%,14 then this suggests a low death rate in the order of 12 deaths per 100,000 visits. A large proportion of deaths (20%) were caused by trauma, of which the majority was accidental. Accidental deaths among travelers have been observed to be increasing http://www.selleckchem.com/products/ipilimumab.html in US citizens and it has been argued that pretravel advice tends to focus on infectious disease risk as opposed to risks that cause injury.22 Personal preparedness and planning is important in increasing safety and decreasing the risk of accidents among travelers who due to unfamiliarity with local conditions

or changed personal behavior are at increased risk of death due to drowning21,25 and car accidents22,26,27; children may be particularly vulnerable.25 In terms of Scottish travelers, it is interesting to note the high proportion of deaths due to selleck screening library circulatory causes (52%), although the proportion is less in this study than that observed by Paixao and colleagues24 at 69%. In that study it was proposed that, among the elderly, deaths abroad may have occurred in their home country had they not traveled. However, our observation that for death due to failure of the circulatory system among those aged 25 to 64, the age at death among those whose bodies were returned for cremation was younger

compared to that of the reference Scottish population, raises the possibility that this difference is linked to travel abroad. A number of factors related to travel abroad may detrimentally affect those with preexisting circulatory conditions including warm climate,28 the journey,29 and lifestyle changes,30 such as increased exertion or changes in diet and/or environmental factors.31 The relationship between age at death Interleukin-3 receptor from cardiovascular disease has been observed among US citizens abroad,32 where 49% of deaths were due to this cause, with the highest proportion of deaths occurring in Western Europe. Cardiovascular death rates among US citizens abroad were found to be higher than among those at home aged 35 to 44. Considering

that many of the travelers died in Southern Europe where the incidence of cardiovascular mortality is much lower than that of Scotland,33 it would be interesting to study at which stage of the journey deaths due to failure in the circulatory system occur. Couch29 noted in an analysis of sudden death due to coronary arteriosclerosis that incidence among visitors was four times that of the local population and suggested that stress due to changing time zones or travel may have contributed. In another study of ischemic heart disease among residents of New York City,34 it was observed that increased deaths due to ischemic heart disease were observed among visitors to that city, while residents away from the city were observed to have lower numbers than expected. This effect was again tentatively linked to stress associated with living in New York for both residents and visitors alike.

New evidence suggests that automatic imitation, otherwise known as ‘imitative compatibility’, shall be considered as a phenomenon that operates independently RO4929097 price from spatial compatibility. So far there are only a few investigations directly aimed at identifying the neural structures dedicated to this process. In the present study, we applied double-pulse transcranial magnetic stimulation (TMS) over the parietal opercula to further investigate the role of these regions in coding imitative compatibility. We found that a temporary disruption of parietal opercula caused the

reduction of the imitative compatibility relative to the sham condition. In particular, the TMS interference with the parietal opercula’s activity modulated the imitative compatibility but not the spatial compatibility, suggesting that these two processes are likely to be independent. “
“The pathological basis of neonatal hypoxia–ischemia (HI) brain damage is characterized by neuronal cell loss.

Oxidative stress is thought to be one of the main causes of HI-induced neuronal cell death. The p38 mitogen-activated protein kinase (MAPK) JAK inhibitor is activated under conditions of cell stress. However, its pathogenic role in regulating the oxidative stress associated with HI injury in the brain is not well understood. Thus, this study was conducted to examine the role of p38 MAPK signaling in neonatal HI brain injury using neonatal rat hippocampal slice cultures exposed to oxygen/glucose deprivation (OGD). Our results indicate that OGD led to a transient increase in p38 MAPK

activation that preceded increases in superoxide generation and neuronal death. This increase in neuronal cell death correlated with an increase in the activation of caspase-3 and the appearance of apoptotic neuronal cells. Pre-treatment of slice cultures with the p38 MAPK inhibitor, SB203580, or the expression of an antisense p38 MAPK construct only in neuronal cells, through a Synapsin I-1-driven adeno-associated virus vector, inhibited p38 MAPK activity and exerted a neuroprotective effect as demonstrated by decreases in OGD-mediated oxidative Sinomenine stress, caspase activation and neuronal cell death. Thus, we conclude that the activation of p38 MAPK in neuronal cells plays a key role in the oxidative stress and neuronal cell death associated with OGD. “
“When viewing the needle of a syringe approaching your skin, anticipation of a painful prick may lead to increased arousal. How this anticipation is reflected in neural oscillatory activity and how it relates to activity within the autonomic nervous system is thus far unknown. Recently, we found that viewing needle pricks compared with Q-tip touches increases the pupil dilation response (PDR) and perceived unpleasantness of electrical stimuli.

01]. Finally, we observed that more hepatotoxic events occurred during the first year of NNRTI therapy compared with the entire period after 1 year (6.6 vs. 2.8 events, respectively, per 100 person-years of treatment; P = 0.04). Long-term NNRTI use was not associated with a higher risk of clinically significant liver toxicity in patients who had been treated with NNRTI for at least 3 years. Following the introduction of highly active antiretroviral therapy SCH727965 chemical structure (HAART), the life expectancy of HIV-infected patients has increased dramatically. In view of the facts that

HAART is a life-long therapy and a successful regimen is intended to be used for many years, the long-term side effects of these antiretroviral drugs are receiving increasing attention. The nonnucleoside reverse transcriptase inhibitors (NNRTIs) efavirenz (EFV) and nevirapine (NVP) are frequently used as components of current antiretroviral regimens. However, NNRTIs are known for their potential to cause hepatotoxicity, which can lead to morbidity and therapy switches. Different studies have reported a cumulative

incidence of severe hepatotoxicity selleck chemicals llc varying from 1.4 to 15.6% in patients treated with NVP [1-5] and from 1.1 to 10% in patients treated with EFV [1-4]. However, the follow-up time in these studies was relatively short, up to 3 years. Data focusing on hepatotoxicity in long-term NNRTI use are scarce [6]. The aim Carnitine palmitoyltransferase II of this retrospective cohort analysis was to evaluate whether the incidence of hepatotoxicity increases with increasing duration of an NNRTI regimen. All HIV-infected patients under follow-up at our clinic until 1 November 2009, who had been receiving an NNRTI-containing HAART regimen for ≥ 3 years, were identified. Patients were included in the analysis if they had continuously used the same NNRTI for a minimum of three years and if at least one serum alanine transaminase (ALT) value per year was available throughout the treatment period. The control group consisted of patients who had exclusively received a protease inhibitor

(PI)-based regimen for at least 3 years and for whom ALT data were available. Demographic, pharmacological and laboratory data at the start of therapy were retrieved from the clinical database and patient records. Patients were considered to have a hepatitis B virus (HBV) infection when HBV DNA and/or the HBV surface antigen (HBsAg) was found at baseline. Hepatitis C virus (HCV) infection was defined as the detection of HCV RNA by polymerase chain reaction. Patients for whom baseline ALT was unknown and those with acute viral hepatitis during NNRTI treatment were excluded from the analysis. Hepatotoxicity was graded according to the modified toxicity scale of the AIDS Clinical Trials Group [1]. Serum ALT values were used rather than serum aspartate aminotransferase (AST) or cholestatic liver enzymes, as ALT is considered to be a more specific marker for liver damage [7].

“
“Serine hydroxymethyltransferase

(SHMT) is a key enzyme in cellular one-carbon pathway and has been studied in many living organisms from bacteria to higher plants and mammals. However, biochemical and molecular characterization of SHMT from photoautotrophic microorganisms remains a challenge. Here, we isolated the SHMT gene from a halotolerant cyanobacterium Aphanothece halophytica (ApSHMT) and expressed it in Escherichia coli. Purified recombinant ApSHMT protein exhibited catalytic reactions for dl-threo-3-phenylserine as well as for l-serine. Catalytic reaction for l-serine was strongly inhibited by NaCl, but not to that level with glycine betaine. Overexpression of ApSHMT in E. coli resulted in the increased accumulation of glycine and serine. Choline and glycine betaine

levels were also significantly Venetoclax find more increased. Under high salinity, the growth rate of ApSHMT-expressing cells was faster compared to its respective control. High salinity also strongly induced the transcript level of ApSHMT in A. halophytica. Our results indicate the importance of a novel pathway; salt-induced ApSHMT increased the level of glycine betaine via serine and choline and conferred the tolerance to salinity stress. Serine is an essential amino acid, and that plays important roles in a variety of biological processes including metabolism, purine and pyrimidine biosynthesis, and generation of activated one-carbon (C-1) unit

(Beaudin et al., 2011). Through serine hydroxymethyltransferase (SHMT), serine associates with glycine metabolism via the glycine decarboxylase complex (GDC). SHMT is a pyridoxal 5′-phosphate (PLP)-dependent many enzyme catalyzing the interconversion of serine and tetrahydrofolate (THF) to glycine and N5, N10-methylene-THF (Schirch et al., 1985). In mammals, SHMT has been shown to be involved in de novo biosynthesis of thymidylate (Anderson & Stover, 2009). Disruption of SHMT increases the risk of neural tube defects (Anderson & Stover, 2009; Beaudin et al., 2011). In prokaryotes such as Escherichia coli, 15% of all carbon atoms assimilated from glucose is estimated to pass through the glycine–serine pathway (Wilson et al., 1993). In plants, SHMT cooperates with the GDC to mediate photorespiratory glycine–serine interconversion (Voll et al., 2005; Bauwe et al., 2010). In cyanobacteria, the SHMT gene was suggested to be essential for cell survival because the complete segregation of SHMT gene could not be generated (Hagemann et al., 2005). Although the enzyme activity of SHMT from a cyanobacterium Synechocystis sp. PCC 6803 has been determined (Eisenhut et al., 2006), molecular properties of cyanobacterial SHMT remain largely unknown. Here, we report on the molecular and biochemical characterization of a putative ApSHMT gene from a halotolerant cyanobacterium Aphanothece halophytica (hereafter called A.

Skin samples of the injected areas were biopsied under local anesthesia

12 and 24 h after injection. All of the animal experiments were approved by the animal research committee at Tokyo University of Agriculture and Technology. Formalin-fixed, paraffin-embedded skin samples were subjected to histopathological analysis. Frozen skin samples were subjected to immunofluorescence analysis for Dsg1 and Dsg3 using a human pemphigus foliaceus serum containing anti-Dsg1 IgG (Amagai et al., 1994, 1999; Ishii et al., 1997) and an AK15 mouse monoclonal antibody against Dsg3 (Tsunoda et al., 2003) (kind gifts from Dr Masayuki Amagai, Keio University School of Medicine, Tokyo, Japan), SB431542 order respectively. The anti-Dsg1 IgG serum and the AK15 monoclonal antibody were detected with fluorescein isothiocyanate-conjugated goat anti-human IgG (MP Biomedicals, Solon, OH) and Alexa Fluor 546 goat anti-mouse IgG (Invitrogen Corp., Carlsbad, CA), respectively. Secreted forms of the recombinant proteins representing the entire extracellular domain of canine Dsg1 (cDsg1) and Dsg3 (cDsg3), fused with the hinge region of human IgG1, an E tag and a His tag at their carboxyl termini, were produced using the baculovirus-expression

system as described previously (Nishifuji et al., 2003a, b). Five insect cell selleck chemicals supernatants containing recombinant cDsg1 or cDsg3 were mixed with 10 μg of purified new ORF protein or PBS alone, and incubated at 37 °C for 12 h. Immunoblotting Cyclooxygenase (COX) with rabbit anti-E-tag polyclonal antibody (Bethyl Laboratories, Montgomery, TX) was performed to detect intact and/or degraded cDsg proteins. PCR was performed with the primers 5′-gcggcatgcctaaaacatatgatgaagccgaa-3′ (forward primer) and 5′-tctctatttacattcagagag-3′ or 5′-tctggatccatcttctgattcagctctttttttcaaa-3′ (reverse primers) to amplify two partial regions of the orf gene. The PCR products were resolved by electrophoresis through a 1.2% (w/v) agarose gel,

and visualized by the application of the SYBR safe DNA gel stain (Invitrogen Corp.). Nucleotide sequence data obtained in this study are available in the DDBJ, EMBL and GenBank nucleotide sequence databases under accession number AB569087. During genome sequencing analysis of S. pseudintermedius strain MS5134, an orf with significant homology to a previously reported ET gene was identified. This orf consisted of 843 bp and was predicted to encode a protein of 279 amino acid residues, including a putative signal peptide in the first 32 amino acids (Fig. 1). The mature protein derived from an orf consisting of 247 amino acid residues with an N-terminal sequence beginning with KTYDEAEIIKK, and a predicted molecular weight and pI of 26.9 kDa and 5.86, respectively. The deduced amino acid sequence of the orf was compared with previously isolated ETs including S. aureus ETs (ETA, ETB and ETD), S.

15), LGN excitatory to L4 inhibitory (P = 0.0619), and TRN inhibitory to LGN excitatory (P = 0.3). The number of neurons in each area is shown in Table 2. The model contained a total of 46 926 neurons and approximately 43 million synapses. Simple and extended versions of the Izhikevich model were used to govern the dynamics of the spiking neurons in this simulation. beta-catenin inhibitor The computational efficiency of these point neurons (single compartment) makes them ideal for large-scale simulations. Izhikevich neurons are also highly realistic and are able to reproduce at least 20 different firing modes seen in the brain, which

include: spiking, bursting, rebound spikes and bursts, subthreshold oscillations, resonance, spike frequency adaptation, spike threshold variability, and bistability of resting and spiking states (Izhikevich, 2004). Inhibitory and excitatory neurons in the cortex were modeled using the simple Izhikevich model, which are described by the following equations Raf inhibitor (Izhikevich, 2003): (2) where v is the membrane potential, u is the recovery variable, I is the input current, and a, b, c and d are parameters chosen based on the neuron type. For regular spiking, excitatory neurons, we set a = 0.01, b = 0.2, c = −65.0 and d = 8.0 (see Fig. 4). For fast-spiking, inhibitory neurons, we set a = 0.1, b = 0.2, c = −65.0 and d = 2.0 (Fig. 4). GABAergic and cholinergic neurons in the BF were modeled as simple

Izhikevich inhibitory and excitatory neurons, respectively. LGN and TRN neurons were modeled using the extended version of the Izhikevich neuron model to account for the bursting and tonic modes of activity, which these neurons have been shown to exhibit (Izhikevich & Edelman, 2008). The equations governing these neurons are given as: (5) The equations for this extended model are similar to the previous model, except they include additional parameters, such as: membrane capacitance (C), resting potential (vr) and instantaneous

threshold potential (vt). For LGN neurons, parameters were set to: a = 0.1, c = −60, d = 10, C = 200, vr = −60 and vt = −50. For TRN neurons, parameters were set to: a = 0.015, c = −55, d = 50, C = 40, vr = −65 and vt = −45 (Izhikevich & Edelman, 2008). To simulate PDK4 the switch between bursting and tonic mode, the b parameter, which is related to the excitability of the cell, was changed depending upon membrane potential, v. Specifically, if v < −65, b was set to 70 and the neuron would be in bursting mode (Fig. 4; bottom, right). If v > −65, b was set to 0 and the neuron would be in tonic mode (Fig. 4; bottom, left). The synaptic input, I, driving each neuron was dictated by simulated AMPA, NMDA, GABAA and GABAB conductances (Izhikevich & Edelman, 2008; Richert et al., 2011). The conductance equations used are well established and have been described in Dayan & Abbott (2001) and Izhikevich et al. (2004). The total synaptic input seen by each neuron was given by: (7) where v is the membrane potential and g is the conductance.

3C. To discern the most stable pattern of cluster assignment across subjects, we applied the spectral clustering algorithm to the consensus matrices and computed the modified silhouette. Figure 3F plots the modified silhouette values, and suggests that, across subjects, the most stable pattern of cluster assignment is for K = 4. Qualitatively, the surface maps for the solutions computed on the basis of the consensus matrix are highly STA-9090 order similar to those computed on the basis of the group-average η2 matrix (Fig. 4), and the VI metric demonstrates that

the best similarity between the clustering solutions is for K = 2 : 4 (Fig. 3G). On the basis of the clustering analyses, we concluded that K = 4 represented the most favorable solution (see Fig. 4). Qualitatively, the four clusters were located in the superior part of the inferior frontal gyrus, bordering the inferior

frontal sulcus (Cluster 1), the lateral pars opercularis Veliparib supplier and pars triangularis (Cluster 2), inferior precentral cortex (Cluster 3) and a fourth region extending medially within the Sylvian fissure from the inferior-most tip of the ventral premotor cortex and the pars opercularis towards the anterior insula (Cluster 4). To verify these clusters as functionally distinct regions of ventrolateral frontal cortex, we examined the RSFC associated with four spherical seed ROIs of 4-mm radius, centered on the centers-of-mass of each of the clusters of the group-average

K = 4 spectral clustering solution. Figure 5 shows the group-level (Z > 2.3; cluster significance P < 0.05, corrected) RSFC for each of the four Meloxicam clusters, as well as direct comparisons between clusters. The pattern of RSFC observed for Cluster 2, which includes the central parts of the pars opercularis and pars triangularis, is very similar to those observed for ROIs based in BAs 44 and 45 (compare Cluster 2 in Fig. 5 with BA 44 and 45 in Fig. 1). Similarly, the pattern of RSFC for Cluster 3, which includes the inferior part of the precentral gyrus, is consistent with that for the ROI based in BA 6 (compare Cluster 3 in Fig. 5 with BA 6 in Fig. 1). The voxels in Cluster 1 probably separate from the rest of the large ventrolateral frontal region of interest that was defined for the clustering analysis by virtue of the fact that they are located along the inferior frontal sulcus on the border with the middle frontal gyrus, which would include voxels of areas 8 and 9/46v in the upper bank of the inferior frontal sulcus and adjacent middle frontal gyrus. Specifically, Cluster 1 exhibited RSFC with almost all of the inferior frontal gyrus, anterior to and including the inferior precentral sulcus, dorsal BA 6 and BA 8 in the middle frontal gyrus, the intraparietal sulcus, and the caudal middle and inferior temporal cortex. The comparison Cluster 1 > Cluster 2 (Fig.

Executive functioning involves the complex cognitive abilities to plan and execute multi-step tasks and process new information and is thought to be impaired

in chronic HIV infection as a result of widespread synaptodendritic injury to frontal-striato-thalamo-cortical brain circuits [17]. Such repair of this synaptodendritic injury may not occur immediately after controlling HIV viraemia with cART, and may explain the observation we have made in our study that executive function improvements occurred later than improvements in the other cognitive domains assessed. Cysique and colleagues have recently described changes in NC function over a 1-year period in 37 HIV-infected www.selleckchem.com/products/ABT-263.html subjects commencing cART, and, similar to our study findings, they observed peak improvements in NC function to occur after 24–36 weeks of therapy [15] with prolonged improvements observed over a 1-year period. However, allocation of cART within this cohort was based on clinician choice, restricting the interpretation of such observations to discern differences between different cART regimens. Also, not all subjects were naïve to cART and all subjects had documented

NC function impairment at baseline. Unlike our study, these Tanespimycin order factors limited the relevance of these observations to HIV-infected neuro-asymptomatic subjects, who represent the majority of HIV-infected subjects commencing cART for the first time. While we have attributed improvements in NC function to the effects of commencing cART, we cannot fully account for confounding factors which may also have resulted in improvements in NC function over the study period. A control arm

within our study allocating subjects not to receive antiretroviral therapy would have strengthened our observations if no improvements 17-DMAG (Alvespimycin) HCl in NC function were observed in subjects allocated to this arm. However, such an approach would not be ethical or feasible as individuals selected to enter the study clinically required to commence antiretroviral therapy. Furthermore, cognitive function is likely to decline over time, rather than improve, and this decline has been reported to be greater in HIV-infected subjects [18], strengthening the argument that the improvements in NC function observed are secondary to commencing cART. Lastly, a learning effect may account for improvements in NC performance. However, all subjects undertook a practice NC test during the study screening period prior to the study baseline test used in our analysis in order to minimize effects of learning on the study results [10], and such effects would not explain the differences in improvements we observed between the study treatment arms.