Topology of the C-Terminal Tail of HIV-1 gp41: Differential Exposure of the Kennedy Epitope on Cell and Viral Membranes

The C-terminal tail (CTT) of the HIV-1 gp41 envelope (Env) protein is increasingly recognized as an important determinant of Env structure and functional properties, including fusogenicity and antigenicity. While the CTT has been commonly referred to as the “intracytoplasmic domain” based on the assumption of an exclusive localization inside the membrane lipid bilayer, early antigenicity studies and recent biochemical analyses have produced a credible case for surface exposure of specific CTT sequences, including the classical “Kennedy epitope” (KE) of gp41, leading to an alternative model of gp41 topology with multiple membrane-spanning domains. The current study was designed to test these conflicting models of CTT topology by characterizing the exposure of native CTT sequences and substituted VSV-G epitope tags in cell- and virion-associated Env to reference monoclonal antibodies (MAbs). Surface staining and FACS analysis of intact, Env-expressing cells demonstrated that the KE is accessible to binding by MAbs directed to both an inserted VSV-G epitope tag and the native KE sequence. Importantly, the VSV-G tag was only reactive when inserted into the KE; no reactivity was observed in cells expressing Env with the VSV-G tag inserted into the LLP2 domain. In contrast to cell-surface expressed Env, no binding of KE-directed MAbs was observed to Env on the surface of intact virions using either immune precipitation or surface plasmon resonance spectroscopy. These data indicate apparently distinct CTT topologies for virion- and cell-associated Env species and add to the case for a reconsideration of CTT topology that is more complex than currently envisioned

The effects of repeated testing, simulated malingering, and traumatic brain injury on high-precision measures of simple visual reaction time

Simple reaction time (SRT), the latency to respond to a stimulus, has been widely used as a basic measure of processing speed. In the current experiments, we examined clinically-relevant properties of a new SRT test that presents visual stimuli to the left or right hemifield at varying stimulus onset asynchronies. Experiment 1 examined test-retest reliability in participants who underwent three test sessions at weekly intervals. In the first test, log-transformed (log-SRT) z-scores, corrected for the influence of age and computer-use, were well predicted by regression functions derived from a normative population of 189 control participants. Test-retest reliability of log-SRT z-scores was measured with an intraclass correlation coefficient (ICC = 0.83) and equaled or exceeded those of other SRT tests and other widely used tests of processing speed that are administered manually. No significant learning effects were observed across test sessions. Experiment 2 investigated the same participants when instructed to malinger during a fourth testing session: 94% showed abnormal log-SRT z-scores, with 83% producing log-SRT z-scores exceeding a cutoff of 3.0, a degree of abnormality never seen in full-effort conditions. Thus, a log-SRT z-score cutoff of 3.0 had a sensitivity (83%) and specificity (100%) that equaled or exceeded that of existing symptom validity tests. We argue that even expert malingerers, fully informed of the malingering-detection metric, would be unable to successfully feign impairments on the SRT test because of the precise control of SRT latencies that would be required. Experiment 3 investigated 26 patients with traumatic brain injury (TBI) tested more than one year post-injury. The 22 patients with mild TBI showed insignificantly faster SRTs than controls, but a small group of four patients with severe TBI showed slowed SRTs. Simple visual reaction time is a reliable measure of processing speed that is sensitive to the effects of malingering and traumatic brain injury

The Effects of Repeated Testing, Simulated Malingering, and Traumatic Brain Injury on Visual Choice Reaction Time

Choice reaction time (CRT), the time required to discriminate and respond appropriately to different stimuli, is a basic measure of attention and processing speed. Here, we describe the reliability and clinical sensitivity of a new CRT test that presents lateralized visual stimuli and adaptively adjusts stimulus onset asynchronies (SOAs) using a staircase procedure. Experiment 1 investigated the test-retest reliability in three test sessions at weekly intervals. Performance in the first test session was accurately predicted from age and computer-use regression functions obtained in a previously studied normative cohort. Central processing time (CentPT), the difference between the CRTs and simple reaction time latencies measured in a separate experiment, accounted for 55% of CRT latency and more than 50% of CRT latency variance. Performance improved significantly across the three test sessions. High intraclass correlation coefficients (ICCs) were seen for CRTs (0.90), CentPTs (0.87), and an omnibus performance measure (0.81) that combined CRT and minimal SOA (mSOA) z-scores. Experiment 2 investigated performance in the same participants when instructed to feign symptoms of traumatic brain injury (TBI): 87% produced abnormal omnibus z-scores. Simulated malingerers showed greater elevations in simple than choice reaction times, and hence reduced CentPTs. Latency-consistency z-scores, based on the difference between the CRTs obtained and those predicted from CentPT latencies, discriminated malingering participants from controls with high sensitivity and specificity. Experiment 3 investigated CRT test performance in military veterans who had suffered combat-related TBI and symptoms of post-traumatic stress disorder and revealed small but significant deficits in performance. The results indicate that the new CRT test shows high test-retest reliability, can assist in detecting participants performing with suboptimal effort, and is sensitive to the effects of TBI on the speed and accuracy of visual processing

A Computerized Test of Design Fluency.

Tests of design fluency (DF) assess a participant's ability to generate geometric patterns and are thought to measure executive functions involving the non-dominant frontal lobe. Here, we describe the properties of a rapidly administered computerized design-fluency (C-DF) test that measures response times, and is automatically scored. In Experiment 1, we found that the number of unique patterns produced over 90 s by 180 control participants (ages 18 to 82 years) correlated with age, education, and daily computer-use. Each line in the continuous 4-line patterns required approximately 1.0 s to draw. The rate of pattern production and the incidence of repeated patterns both increased over the 90 s test. Unique pattern z-scores (corrected for age and computer-use) correlated with the results of other neuropsychological tests performed on the same day. Experiment 2 analyzed C-DF test-retest reliability in 55 participants in three test sessions at weekly intervals and found high z-score intraclass correlation coefficients (ICC = 0.79). Z-scores in the first session did not differ significantly from those of Experiment 1, but performance improved significantly over repeated tests. Experiment 3 investigated the performance of Experiment 2 participants when instructed to simulate malingering. Z-scores were significantly reduced and pattern repetitions increased, but there was considerable overlap with the performance of the control population. Experiment 4 examined performance in veteran patients tested more than one year after traumatic brain injury (TBI). Patients with mild TBI performed within the normal range, but patients with severe TBI showed reduced z-scores. The C-DF test reliably measures visuospatial pattern generation ability and reveals performance deficits in patients with severe TBI

The effects of repeat testing, malingering, and traumatic brain injury on visuospatial memory span

Spatial span tests such as the Corsi Block Test (CBT) and the spatial span test of the Wechsler Memory Scale are widely used to assess deficits in spatial working memory. We conducted three experiments to evaluate the test-retest reliability and clinical sensitivity of a new computerized spatial span test (C-SST) that incorporates psychophysical methods to improve the precision of spatial span measurement. In Experiment 1, we analyzed C-SST test-retest reliability in 49 participants who underwent three test sessions at weekly intervals. Intraclass correlation coefficients (ICC) were higher for a psychophysically derived mean span (MnS) metric (0.83) than for the maximal span and total correct metrics used in traditional spatial-span tests. Response times (ReTs) also showed high ICCs (0.93) that correlated negatively with MnS scores and correlated positively with response-time latencies from other tests of processing speed. Learning effects were insignificant. Experiment 2 examined the performance of Experiment 1 participants when instructed to feign symptoms of traumatic brain injury: 57% showed abnormal MnS z-scores. A MnS z-score cutoff of 3.0 correctly classified 36% of simulated malingerers and 91% of the subgroup of 11 control participants with abnormal spans. Malingerers also made more substitution errors than control participants with abnormal spans (sensitivity = 43%, specificity = 91%). In addition, malingerers showed no evidence of ReT slowing, in contrast to significant abnormalities seen on other malingered tests of processing speed. As a result, differences between ReT z-scores and z-scores on other processing speed tests showed very high sensitivity and specificity in distinguishing malingering and control participants with either normal or abnormal spans. Experiment 3 examined C-SST performance in a group of patients with predominantly mild traumatic brain injury (TBI): neither MnS nor ReT z-scores showed significant group-level abnormalities. The C-SST improves the reliability and sensitivity of spatial span testing, can accurately detect malingering, and shows that visuospatial working memory is largely preserved in patients with predominantly mild TBI

Age-related slowing of response selection and production in a visual choice reaction time task

Aging is associated with delayed processing in choice reaction time (CRT) tasks, but the processing stages most impacted by aging have not been clearly identified. Here, we analyzed CRT latencies in a computerized serial visual feature-conjunction task. Participants responded to a target letter (probability 40%) by pressing one mouse button, and responded to distractor letters differing either in color, shape, or both features from the target (probabilities 20% each), by pressing the other mouse button. Stimuli were presented randomly to the left and right visual fields and stimulus onset asynchronies (SOAs) were adaptively reduced following correct responses using a staircase procedure. In Experiment 1, we tested 1466 participants who ranged in age from 18 to 65 years. CRT latencies increased significantly with age (r = 0.47, 2.80 ms/year). Central processing time (CPT), isolated by subtracting simple reaction times (obtained in a companion experiment performed on the same day) from CRT latencies, accounted for more than 80% of age-related CRT slowing, with most of the remaining increase in latency due to slowed motor responses. Participants were faster and more accurate when the stimulus location was spatially compatible with the mouse button used for responding, and this effect increased slightly with age. Participants took longer to respond to distractors with target color or shape than to distractors with no target features. However, the additional time needed to discriminate the more target-like distractors did not increase with age. In Experiment 2, we replicated the findings of Experiment 1 in a second population of 178 participants (ages 18-82 years). CRT latencies did not differ significantly in the two experiments, and similar effects of age, distractor similarity, and stimulus-response spatial compatibility were found. The results suggest that the age-related slowing in visual CRT latencies is largely due to delays in response selection and production