Coxiella burnetii NMII infections were initiated as described and fixed at 0, 8, 16, 24, 48, 96, and 168 hpi with 4% paraformaldehyde, 0.05% Tween-20 in phosphate-buffered saline for 15 min at room temperature. Indirect immunofluorescent antibody (IFA) analysis was performed by dual staining as described previously (Morgan et al., 2010). Micrograph images were captured via a Nikon DS FI1 camera on a Nikon Eclipse TE 2000-S microscope at × 400 magnification, with nis-elements f 3.00 software. A magnification of × 400 was used as opposed to × 600, as used previously (Morgan et al., 2010). Micrograph capture settings were uniform for all images. Using a modification of a method used previously in C.

burnetii studies that uses relative pixel ratios in sample quantitation (Zamboni et al., 2001), each micrograph image was analyzed using imagej version RG7422 1.42n (Wayne Rasband, NIH) software. Five fields of view from each time sampled (three biological samples of each) were digitally captured. The matching Alexa Fluor® 555 and Alexa Fluor® 488 images were stacked (paired) and converted to gray scale (8 bit). No fewer than five regions of interest (ROI) were blindly selected from each field of view of the 555 nm grayscale images. This provided at least 75 ROIs for each time point analyzed. Saturated regions of an image were not selected and ROI size varied

Epacadostat clinical trial depending on the PV size. The pixel densities within the identical ROIs from each stacked image were then measured

as published previously (Collins, 2007). The mean pixel densities were then compared to obtain the 488 : 555 ratio for each ROI. These individual ratios were then averaged (≥75 individual ratios/time point) to determine the relative expression of IcmT to whole C. burnetii NMII. The final 488 : 555 (IcmT : C. Histidine ammonia-lyase burnetii) ratio for each time point was then divided into the 0 hpi ratio to obtain the final IcmT relative expression levels. The statistical significance between each time point was evaluated using single-factor anova with a 95% confidence interval using ms excel 2007 (Microsoft). The C. burnetii T4BSS RI gene linkage map suggests that three groups of transcriptionally linked genes exist (see Fig. 1a). These include: (1) icmXCBU1651icmW, (2) icmVdotACBU1647, and (3) CBU1646dotBdotCdotDicmSicmT. To demonstrate transcriptional linkage between the genes within each group, RT-PCR analysis was performed using oligonucleotide primers (see Table 1) designed to span intergenic sequences and/or adjoining ORFs. The diamond-ended lines in Fig. 1a indicate the position of primers and DNA products that would result from RT-PCR amplification. Using total RNA harvested from Vero cells infected with C. burnetii NMII as a template, amplification products were observed (Fig. 1b) for each linkage region: (1) icmW–icmX, (2) icmV–dotA, dotA–CBU1647, and (3) icmT–dotD, dotD–dotB, dotB–CBU1646 (Fig. 1b).

Coxiella burnetii NMII infections were initiated as described and fixed at 0, 8, 16, 24, 48, 96, and 168 hpi with 4% paraformaldehyde, 0.05% Tween-20 in phosphate-buffered saline for 15 min at room temperature. Indirect immunofluorescent antibody (IFA) analysis was performed by dual staining as described previously (Morgan et al., 2010). Micrograph images were captured via a Nikon DS FI1 camera on a Nikon Eclipse TE 2000-S microscope at × 400 magnification, with nis-elements f 3.00 software. A magnification of × 400 was used as opposed to × 600, as used previously (Morgan et al., 2010). Micrograph capture settings were uniform for all images. Using a modification of a method used previously in C.

burnetii studies that uses relative pixel ratios in sample quantitation (Zamboni et al., 2001), each micrograph image was analyzed using imagej version learn more 1.42n (Wayne Rasband, NIH) software. Five fields of view from each time sampled (three biological samples of each) were digitally captured. The matching Alexa Fluor® 555 and Alexa Fluor® 488 images were stacked (paired) and converted to gray scale (8 bit). No fewer than five regions of interest (ROI) were blindly selected from each field of view of the 555 nm grayscale images. This provided at least 75 ROIs for each time point analyzed. Saturated regions of an image were not selected and ROI size varied

GSK 3 inhibitor depending on the PV size. The pixel densities within the identical ROIs from each stacked image were then measured

as published previously (Collins, 2007). The mean pixel densities were then compared to obtain the 488 : 555 ratio for each ROI. These individual ratios were then averaged (≥75 individual ratios/time point) to determine the relative expression of IcmT to whole C. burnetii NMII. The final 488 : 555 (IcmT : C. Alanine-glyoxylate transaminase burnetii) ratio for each time point was then divided into the 0 hpi ratio to obtain the final IcmT relative expression levels. The statistical significance between each time point was evaluated using single-factor anova with a 95% confidence interval using ms excel 2007 (Microsoft). The C. burnetii T4BSS RI gene linkage map suggests that three groups of transcriptionally linked genes exist (see Fig. 1a). These include: (1) icmXCBU1651icmW, (2) icmVdotACBU1647, and (3) CBU1646dotBdotCdotDicmSicmT. To demonstrate transcriptional linkage between the genes within each group, RT-PCR analysis was performed using oligonucleotide primers (see Table 1) designed to span intergenic sequences and/or adjoining ORFs. The diamond-ended lines in Fig. 1a indicate the position of primers and DNA products that would result from RT-PCR amplification. Using total RNA harvested from Vero cells infected with C. burnetii NMII as a template, amplification products were observed (Fig. 1b) for each linkage region: (1) icmW–icmX, (2) icmV–dotA, dotA–CBU1647, and (3) icmT–dotD, dotD–dotB, dotB–CBU1646 (Fig. 1b).

Third, if two CYC202 price conditionally dependent findings are entered, only the one with the highest positive likelihood ratio is accounted for. Finally, at every step, the sum of all probabilities is reset at 100%. At any time during the consultation, the user can also ask the help of the tutor module that lists the relevant findings to explore, or suggests step-by-step further testing, with reassessment

of the case each time a new finding is entered (“wizard” button). The tutor will not end the case before the probability of a diagnosis is considered high enough by the system (over the treatment threshold), before all relevant excluders for this disease have PKC inhibitor been exhausted, and before

competing dangerous and treatable diagnoses are sufficiently excluded. Following this study and coinvestigator’s suggestions, KABISA TRAVEL has been upgraded recently, but with no major modifications. The software is now freely accessible at www.kabisa.be: KABISA V; setting “Travel clinic”; module “Expert. A first single-center retrospective study has evaluated the KABISA TRAVEL in 54 febrile travelers presenting at a Belgian emergency ward and demonstrated that 93% of the cases were correctly diagnosed.12 The present study intended to assess prospectively the diagnostic accuracy of the KABISA TRAVEL in different European settings dealing with travel-related pathology, and to compare it to travel physicians’ performances. Secondary objectives were to evaluate the clinical utility of the KABISA TRAVEL software and (-)-p-Bromotetramisole Oxalate the specific contribution of the tutor. From December 2007 to April 2009, travelers with fever after a stay in the tropics were included prospectively in a multicenter trial conducted in 10 referral travel clinics located in the Netherlands, Italy, Spain, and Belgium (nine tertiary referral hospitals with

travel clinics and one outpatient referral travel clinic). Anonymous data from all collaborating centers were centralized and analyzed at the Institute of Tropical Medicine, Antwerp, Belgium. We prospectively enrolled patients of any age presenting at one of the study centers with ongoing fever occurring within 3 months after a stay in the tropics. Ongoing fever was defined by an axillary temperature of 38°C or higher, documented by the patient or a physician whenever in the past 3 days before the first consultation. Tropics and subtropics corresponded to all countries at least partly situated between the 35°-northern and 35°-southern latitude, except the United States, European countries, Japan, and Australia. The study patients were clinically managed by each coinvestigator (all of them being physicians with expertise in travel medicine) according to the usual standard of care in each site/country.

The spyder output, which consists

of a text file summarizing the location of indels and substitutions, was used to identify locations where degeneracies could be introduced to compensate for common mismatches. A second analysis using RDP Probe Match was used to evaluate the new primer and verify that it did not compromise specificity (Table 4). oligocalc confirmed primer quality, including a suitable GC content, and the absence of self-complementarity, hairpins, and 3′- primer–primer complementarity (data not shown). The most substantial improvements http://www.selleckchem.com/products/pf-562271.html were for the primers targeting Alphaproteobacteria (Alf28f), Gammaproteobacteria (Gamma395f), Bacteriodetes (CFB555f), Firmicutes (Firm350f), and Archaea (A571F) resulting in 22%, 42%, 15%, 18%, and 26% increases in coverage, respectively, while nonspecific mismatches remained low (0.03–2.56%) (Table 4). Analysis of primers designed using arb and primrose (i.e. those designed by Muhling et al., 2008) by spyder indicated that these primers could be improved without sacrificing specificity by adding targeted degeneracies (Table 4). This may be because current databases are more comprehensive and/or that arb does not include a feature for including degeneracies in primer design (Muhling et al., 2008). spyder also identified improvements (5.9% increase) of the commonly

used eubacterial primer F27, which is the forward primer used along with R1492 for the Human Microbiome

Project Sanger sequencing libraries CX-5461 price (Turnbaugh et al., 2007). The F27 primer was also the forward primer of choice in the recent survey of the microbiota of the oral cavity of healthy adults in which over 10 000 full-length 16S rRNA gene sequences were analyzed (Bik et al., 2010). Methocarbamol In the majority of cases, spyder determined that only the forward or the reverse primer of a standard set could be improved. The lack of nonspecific hits associated with the improved primer indicates that it may be beneficial to use a comprehensive universal or alternate primer to complete the pair in the event that the current primer pair possesses differential coverage. Adding degeneracies is a common method for improving primers; however, it is possible that too many degenerate sites will diminish the primers target specificity. As such, other methods to increase mismatch tolerance should also be considered such as using long primers (25+ bases long), increasing dNTP concentrations, MgCl2, and annealing time, as well as using annealing temperatures below the Tm of the primers (Kwok et al., 1994). PCR cycle number should also be minimized along with the pooling of multiple PCR products to reduce the high variation that is inherent in the early stages of multitemplate PCR (Brooks et al., 2007).

In contrast, HU-210 administration to N-methyl-D-aspartate receptor knockdown mice was ineffective

at promoting striatal ERK1/2 inactivation. Genetic deletion of other potential ERK1/2 mediators, the dopamine D2 receptors or β-arrestin-1 or -2, did not affect the HU-210-induced modulation of ERK1/2 signaling in the striatum. These results support the hypothesis that dopamine D1 receptors and N-methyl-D-aspartate receptors CFTR modulator act in an opposite manner to regulate striatal CB1 cannabinoid receptor signal transduction. “
“In a three-dimensional (3D) world most saccades are made towards visual targets that are located at different distances. We previously demonstrated that gaze shifts within 3D space consist of two stages: a target saccade followed by a corrective saccade during gaze fixation that directs the eyes

to the physical target location. We proposed that, by accurately positioning the eyes on the visual object, the visual system maintains an orderly representation of the visual world. In this study we used a double saccade experiment to assess the function of corrective saccades in humans. We found that, when a corrective eye movement occurred during fixation on the first target point, the direction of the second saccade towards the next target point was accurate. When a corrective saccade was absent, a directional error of the second target saccade was observed. This finding, which cannot be explained by current models of eye movement control, supports the idea of a two-step model in saccade phosphatase inhibitor library programming. We suggest that the motor system sends a corollary discharge when programming a corrective saccade for maintaining an orderly representation of the visual world. In conclusion, our results indicate that corrective saccades have a role in programming target saccades within 3D space. “
“Surround inhibition is a physiological mechanism that is hypothesised to improve contrast between signals in the central nervous system. In the human motor

system, motor surround inhibition (mSI) can be assessed using transcranial magnetic Cyclic nucleotide phosphodiesterase stimulation (TMS). We evaluated whether it is possible to modulate mSI, using a paradigm able to induce plastic effects in primary motor cortex (M1). Fifteen healthy volunteers participated in the experiments. To assess mSI, we delivered single pulses at rest and at the onset of a right thumb abduction. TMS pulses over abductor digiti minimi (ADM; surround muscle) hotspot were delivered when EMG activity in right abductor pollicis brevis (APB; active muscle) > 100 μV was detected. Paired associative stimulation (PAS) was delivered using peripheral median nerve electric stimulation and TMS over APB M1 area at an interstimulus interval of 21.5 ms for the real PAS (PAS21.5) and 100 ms for the sham PAS (PAS100). To verify the effect of PAS21.

In addition, overexpression of glycerol-3-phosphate dehydrogenase (glpD) and glycerol-3-phosphate acyltransferase (plsB) involved in energy metabolism (Spoering et al., 2006) and overexpression of relA involved in ppGpp synthesis (Korch et al., 2003) also caused increased persister Bleomycin manufacturer formation. We have recently identified a new persister gene phoU, previously identified as a repressor of phosphate uptake system pstSCAB, in E. coli using a transposon mutagenesis approach (Li & Zhang, 2007). Mutation in PhoU leads to a generalized higher susceptibility than the parent strain to a diverse range of antibiotics and stresses and a defect in persister formation.

Microarray studies indicated that PhoU mutant surprizingly expressed high levels of energy metabolism genes, transporter genes and flagella and chemotaxis genes, which suggests that PhoU is a global repressor for cellular metabolism and its inactivation leads to a hyperactive metabolic state

(Li & Zhang, 2007). We thus proposed a new model of persister formation based on PhoU as a global negative regulator, which suppresses the cellular metabolism of the bacteria by downregulating or shutting down the genes or proteins involved in energy production, membrane transport, AZD9291 ic50 etc., to facilitate persister formation (Li & Zhang, 2007). However, persisters are not homogeneous (Zhang, 2004) and are most likely mediated by multiple mechanisms. To shed further light on possible new persister mechanisms, in this study, taking advantage of our successful experience of screening a transposon mutant library (Li & Zhang, 2007), we screened the E. coli Keio deletion mutant library and identified two new persister genes sucB and ubiF involved in energy metabolism, whose inactivation caused a defect in persister survival as demonstrated by higher susceptibility to different antibiotics and stresses than the parent strain. Ampicillin, norfloxacin, gentamicin, trimethoprim, tetracycline,

kanamycin and chloramphenicol were obtained from Sigma-Aldrich Chemical Co., and their stock solutions were freshly prepared, filter-sterilized and used at appropriate concentrations pentoxifylline as indicated. Escherichia coli K-12 parent strain BW25113 and its isogenic deletion mutant library of the Keio collection (Baba et al., 2006) were used for the genetic screens to identify mutants with a defect in persister survival after antibiotic exposure. Escherichia coli cells were routinely grown in Luria–Bertani (LB) medium. For sucB and ubiF mutants, 30 μg mL−1 kanamycin was used, and for complementation of sucB and ubiF mutants, 30 μg mL−1 kanamycin and 30 μg mL−1 chloramphenicol were added. M9 minimal medium with a final concentration of 0.4% glucose and 0.05% magnesium sulfate was used as a nutrient-limiting medium. Saline (0.9% sodium chloride) was used as a condition for the starvation experiment. M9 minimal medium at pH 3.0 and 5.

In addition, overexpression of glycerol-3-phosphate dehydrogenase (glpD) and glycerol-3-phosphate acyltransferase (plsB) involved in energy metabolism (Spoering et al., 2006) and overexpression of relA involved in ppGpp synthesis (Korch et al., 2003) also caused increased persister Sirolimus formation. We have recently identified a new persister gene phoU, previously identified as a repressor of phosphate uptake system pstSCAB, in E. coli using a transposon mutagenesis approach (Li & Zhang, 2007). Mutation in PhoU leads to a generalized higher susceptibility than the parent strain to a diverse range of antibiotics and stresses and a defect in persister formation.

Microarray studies indicated that PhoU mutant surprizingly expressed high levels of energy metabolism genes, transporter genes and flagella and chemotaxis genes, which suggests that PhoU is a global repressor for cellular metabolism and its inactivation leads to a hyperactive metabolic state

(Li & Zhang, 2007). We thus proposed a new model of persister formation based on PhoU as a global negative regulator, which suppresses the cellular metabolism of the bacteria by downregulating or shutting down the genes or proteins involved in energy production, membrane transport, Vincristine concentration etc., to facilitate persister formation (Li & Zhang, 2007). However, persisters are not homogeneous (Zhang, 2004) and are most likely mediated by multiple mechanisms. To shed further light on possible new persister mechanisms, in this study, taking advantage of our successful experience of screening a transposon mutant library (Li & Zhang, 2007), we screened the E. coli Keio deletion mutant library and identified two new persister genes sucB and ubiF involved in energy metabolism, whose inactivation caused a defect in persister survival as demonstrated by higher susceptibility to different antibiotics and stresses than the parent strain. Ampicillin, norfloxacin, gentamicin, trimethoprim, tetracycline,

kanamycin and chloramphenicol were obtained from Sigma-Aldrich Chemical Co., and their stock solutions were freshly prepared, filter-sterilized and used at appropriate concentrations fantofarone as indicated. Escherichia coli K-12 parent strain BW25113 and its isogenic deletion mutant library of the Keio collection (Baba et al., 2006) were used for the genetic screens to identify mutants with a defect in persister survival after antibiotic exposure. Escherichia coli cells were routinely grown in Luria–Bertani (LB) medium. For sucB and ubiF mutants, 30 μg mL−1 kanamycin was used, and for complementation of sucB and ubiF mutants, 30 μg mL−1 kanamycin and 30 μg mL−1 chloramphenicol were added. M9 minimal medium with a final concentration of 0.4% glucose and 0.05% magnesium sulfate was used as a nutrient-limiting medium. Saline (0.9% sodium chloride) was used as a condition for the starvation experiment. M9 minimal medium at pH 3.0 and 5.

In

contrast, the glial scar, evaluated by glial fibrillary acidic protein staining, showed its highest intensity 21 Selleck Tacrolimus days post-injury in both models. The number of apoptotic oligodendrocytes, detected by CC1/caspase-3 co-labeling, was increased in both models in all evaluated regions. Finally, the numbers of OPCs, evaluated with the markers Tcf4 and Olig2, were increased from day 2 (Olig2) or day 7 (Tcf4) post-injury (P ≤ 0.05). Our results indicate that TBI induces oligodendrocyte apoptosis and widespread myelin loss, followed by a concomitant increase in the number of OPCs. Prevention of myelin loss and oligodendrocyte death may represent novel therapeutic targets for TBI. “
“Working memory (WM) performance in humans can be improved by structuring and organizing the material to be remembered. For visual and verbal information, this process of structuring has been associated with the involvement of a prefrontal–parietal network, but for non-verbal auditory material, the brain areas that facilitate WM for structured information have remained elusive. Using functional magnetic resonance imaging, this study compared neural correlates underlying encoding and rehearsal of auditory WM for structured and unstructured material.

Musicians and non-musicians performed a WM task on five-tone sequences that were either tonally structured (with all tones Selleck Obeticholic Acid belonging to one tonal key) or tonally unstructured (atonal) sequences. Functional differences were observed for musicians (who are experts in the music domain), but not for non-musicians – The right

pars orbitalis was activated more strongly in musicians during the encoding of unstructured (atonal) vs. structured (tonal) sequences. In addition, data for musicians showed that a lateral (pre)frontal–parietal network (including the right premotor cortex, right inferior precentral sulcus and left intraparietal sulcus) was activated during WM rehearsal of structured, as compared with unstructured, sequences. Our findings indicate that this network plays a role in strategy-based WM for non-verbal auditory information, corroborating previous results Aldehyde dehydrogenase showing a similar network for strategy-based WM for visual and verbal information. “
“Parkinson’s disease is most commonly modelled via unilateral infusion of the neurotoxin 6-hydroxydopamine (6-OHDA) in the rat, but recent work has been aimed to translate the reproducibility and reliability of the model to the mouse. Here we present the effects of unilateral 6-OHDA lesions to either the medial forebrain bundle or the substantia nigra (SN) in mice, which were trained on a lateralised choice reaction time (RT) task.

To validate these observations, we experimentally converted the mutL gene between the wild-type and 6bpΔmutL in S. typhimurium and inspected the bacterial mutability status. When 6bpΔmutL was converted

to mutL, the originally highly mutable Salmonella strains regained genetic stability; when mutL was converted to 6bpΔmutL, the mutability was elevated 100-fold. Interestingly, Nutlin-3a manufacturer mutL cells were found to grow out of 6bpΔmutL cells; the new mutL cells eventually replaced the original 6bpΔmutL population. As conversion between mutL and 6bpΔmutL may occur readily during DNA replication, it may represent a previously unrecognized mechanism to modulate bacterial mutability at the population level, allowing bacteria to respond rapidly to changing environments while minimizing the risks associated with persistent hypermutability. Previously, we observed the peculiar phenomenon of genome diversification, i.e. mutants of Salmonella check details typhimurium LT7 stored at room temperature kept changing the physical structure of the genome (Liu et al., 2003). We have since concentrated on identifying the genetic basis responsible for this phenomenon, with a focus on mismatch repair (MMR) genes including mutL, mutS and mutH.

Our initial work showed that all screened S. typhimurium LT7 mutants had intact mutS and mutH genes, but a deletion was found in mutL; this genotype was designated 6bpΔmutL (Gong et al., 2007). MutL has been suggested to function as a master coordinator or molecular matchmaker in the MMR system. It has a weak ATPase Demeclocycline function, binds to DNA, interacts directly with MutS, MutH and UvrD, and is required for initiation as well as subsequent steps in MMR processes (Ban & Yang, 1998; Hall et al., 1998; Ban et al., 1999; Spampinato & Modrich, 2000). The deleted sequence that we identified in mutL is one of three tandem 6-bp

repeats, GCTGGC GCTGGC GCTGGC. Similar repeats were reported in Escherichia coli, although they were presented as G CTGGCG CTGGCG CTGGCG (Shaver & Sniegowski, 2003). The sequence (G)CTGGC GCTGGC GCTGGC C in Salmonella and (G) CTGGCG CTGGCG CTGGCG in E. coli both code for the amino acid sequence LALALA, which lies in a region of MutL that forms the lid of the ATP-binding pocket (Ban & Yang, 1998; Ban et al., 1999; Yang, 2000; Yang et al., 2000). In our case, the deletion of one of the 6-bp repeats will lead to one of the three LA sets missing in the ATP lid structure of MutL and may thus impair ATPase activity. As the repeat structure would facilitate deletion or duplication via slipped-strand mispairing (Streisinger et al., 1966; Levinson & Gutman, 1987), one can imagine that MMR may cease functioning when the 6-bp repeats of mutL decrease or increase one or more copies and resume functioning when the copy number again becomes three, both by ‘errors’ during replication.

11 and 15%, respectively; P=0.0001), heterosexual (56, 16 and 20%, respectively; P=0.0001) and Black African (45, 9 and 13%, respectively;

P=0.0001) than either late starters or ideal starters. As would be expected from the way the groups were defined, there was a significantly shorter time between first presentation and starting HAART for late presenters compared with the other two groups (medians 0.1, 4.9 and 3.3 years, respectively; P=0.0001). Median follow-up after starting see more HAART was slightly longer among late starters (median 3.6 years) than either late presenters (3.4 years) or ideal starters (2.9 years; P=0.0001). In terms of initial regimen, the majority of patients in all groups started two NRTIs with an NNRTI (Table 1). The proportions of late presenters, late starters and ideal starters commencing a boosted PI-based regimen were 21, 19 and 17%, respectively (P=0.003). Patient disposition at 48 and 96 weeks is described in Figure 1. By 48 weeks, 86.4, 88.0 and 80.4% of late presenters, late starters and ideal starters remained under follow-up, respectively (P=0.0001). Most were receiving antiretroviral therapy (81.1% of all patients and 95.3% of those remaining under follow-up, with no major differences between late presenters and late starters) and 81.7 and 84.9% of those

on antiretroviral therapy had a viral load and CD4 cell count measurement, respectively, Avelestat (AZD9668) BYL719 within the week 40–56 window. By 96 weeks, 82.2, 83.2 and 81.5% of late presenters, late starters and ideal starters, respectively, who were alive and under follow-up at week 48 remained under follow-up (P=0.63). Again, most were on antiretroviral therapy (77.8% of those under

follow-up at week 48; 94.6% of those alive and under follow-up at week 96), and 81.0 and 83.1% of those on antiretroviral therapy at this time had a viral load and CD4 cell count in the week 88–104 window, respectively. Proportions with viral suppression to <50 copies/mL at 48 weeks were 82.4% for late presenters, 85.5% for late starters and 89.3% for ideal starters (P=0.0001). By multivariable analysis (adjusted for gender, mode of infection, ethnicity, age, calendar year, AIDS status and initial regimen), the difference between virological outcomes in late presenters and late starters was not significant at week 48, although there was a marginally nonsignificant difference in virological outcome between late starters and ideal starters (Table 2); by 96 weeks, the differences were further reduced and remained nonsignificant. The median CD4 cell count increase at 48 weeks was significantly lower for late presenters (161 cells/μL) than for late starters (206 cells/μL); while there remained a significant difference in CD4 count increase between the two groups at 96 weeks, this difference was reduced.