Endovascular Brain-Computer Interfaces in Poststroke Paralysis.

Stroke is a leading cause of paralysis, most frequently affecting the upper limbs and vocal folds. Despite recent advances in care, stroke recovery invariably reaches a plateau, after which there are permanent neurological impairments. Implantable brain-computer interface devices offer the potential to bypass permanent neurological lesions. They function by (1) recording neural activity, (2) decoding the neural signal occurring in response to volitional motor intentions, and (3) generating digital control signals that may be used to control external devices. While brain-computer interface technology has the potential to revolutionize neurological care, clinical translation has been limited. Endovascular arrays present a novel form of minimally invasive brain-computer interface devices that have been deployed in human subjects during early feasibility studies. This article provides an overview of endovascular brain-computer interface devices and critically evaluates the patient with stroke as an implant candidate. Future opportunities are mapped, along with the challenges arising when decoding neural activity following infarction. Limitations arise when considering intracerebral hemorrhage and motor cortex lesions; however, future directions are outlined that aim to address these challenges.

Post-stimulus beta responses are modulated by task duration.

Modulation of beta-band neural oscillations during and following movement is a robust marker of brain function. In particular, the post-movement beta rebound (PMBR), which occurs on movement cessation, has been related to inhibition and connectivity in the healthy brain, and is perturbed in disease. However, to realise the potential of the PMBR as a biomarker, its modulation by task parameters must be characterised and its functional role determined. Here, we used MEG to image brain electrophysiology during and after a grip-force task, with the aim to characterise how task duration, in the form of an isometric contraction, modulates beta responses. Fourteen participants exerted a 30% maximum voluntary grip-force for 2, 5 and 10 s. Our results showed that the amplitude of the PMBR is modulated by task duration, with increasing duration significantly reducing PMBR amplitude and increasing its time-to-peak. No variation in the amplitude of the movement related beta decrease (MRBD) with task duration was observed. To gain insight into what may underlie these trial-averaged results, we used a Hidden Markov Model to identify the individual trial dynamics of a brain network encompassing bilateral sensorimotor areas. The rapidly evolving dynamics of this network demonstrated similar variation with task parameters to the 'classical' rebound, and we show that the modulation of the PMBR can be well-described in terms of increased frequency of beta events on a millisecond timescale rather than modulation of beta amplitude during this time period. Our results add to the emerging picture that, in the case of a carefully controlled paradigm, beta modulation can be systematically controlled by task parameters and such control can reveal new information as to the processes that generate the average beta timecourse. These findings will support design of clinically relevant paradigms and analysis pipelines in future use of the PMBR as a marker of neuropathology.

Modulation of post-movement beta rebound by contraction force and rate of force development.

Movement induced modulation of the beta rhythm is one of the most robust neural oscillatory phenomena in the brain. In the preparation and execution phases of movement, a loss in beta amplitude is observed [movement related beta decrease (MRBD)]. This is followed by a rebound above baseline on movement cessation [post movement beta rebound (PMBR)]. These effects have been measured widely, and recent work suggests that they may have significant importance. Specifically, they have potential to form the basis of biomarkers for disease, and have been used in neuroscience applications ranging from brain computer interfaces to markers of neural plasticity. However, despite the robust nature of both MRBD and PMBR, the phenomena themselves are poorly understood. In this study, we characterise MRBD and PMBR during a carefully controlled isometric wrist flexion paradigm, isolating two fundamental movement parameters; force output, and the rate of force development (RFD). Our results show that neither altered force output nor RFD has a significant effect on MRBD. In contrast, PMBR was altered by both parameters. Higher force output results in greater PMBR amplitude, and greater RFD results in a PMBR which is higher in amplitude and shorter in duration. These findings demonstrate that careful control of movement parameters can systematically change PMBR. Further, for temporally protracted movements, the PMBR can be over 7 s in duration. This means accurate control of movement and judicious selection of paradigm parameters are critical in future clinical and basic neuroscientific studies of sensorimotor beta oscillations. Hum Brain Mapp 37:2493-2511, 2016. © 2016 Wiley Periodicals, Inc.

Prolonged infrapatellar tendon vibration does not influence quadriceps maximal or explosive isometric force production in man.

PURPOSE: The influence of muscle/tendon vibration on maximal muscle performance is unclear. This study examined the effect of a prolonged tendon vibration stimulus on maximum voluntary contraction (MVC) and explosive voluntary contraction (EVC) performance. METHODS: Eighteen young healthy males (nine strength trained and nine untrained) completed a series of isometric unilateral knee extensions (EVCs, electrically evoked octet responses, MVCs, ramp contractions) pre and post two separate 30-min intervention trials; infrapatellar tendon vibration (80 Hz), and quiet sitting (control). H max and M max were measured at the start and end of each series of contractions, both pre- and post-intervention (i.e., at four time points). Knee extensor force and both quadriceps and hamstrings EMG were measured throughout each series of contractions. RESULTS: Vibration had no effect on either maximum force (ANOVA, trial × time interaction P = 0.92), explosive force (P ≥ 0.36), or the associated agonist EMG amplitude during these tasks (P ≥ 0.23). Octet responses were also unaffected by vibration (P ≥ 0.39). Conversely, post-intervention H max/M max was 60 % lower in the vibration trial vs. control, and remained 38 % lower at the end of the post-intervention measurements (t test, both P < 0.01). Individual H max/M max depression did not correlate to changes in either maximum or explosive force (Spearman's Rank, P ≥ 0.54), and training status had no influence on the effect of vibration. CONCLUSION: Prolonged infrapatellar tendon vibration depressed H-reflex amplitude, but did not affect either maximal or explosive isometric force production of the quadriceps.

Comparison of Hemodynamic Brain Responses Between Big Wave Surfers and Non-big Wave Surfers During Affective Image Presentation.

Background: Big wave surfers are extreme sports athletes who expose themselves to life-threatening risk when training and competing. Little is known about how and why extreme sports athletes choose to participate in their chosen sports. This exploratory study investigated potential neurophysiological and psychometric differences between big and non-big wave surfers. Methods: Thirteen big wave surfers (BWS) and 10 non-big wave surfers (CON) viewed a series of images from the International Affective Picture System (IAPS) while undergoing brain functional magnetic resonance imaging (fMRI). The Fear Schedule Survey-III, Arnett Inventory of Sensation Seeking, Discrete Emotions Questionnaire, and Positive and Negative Affect Schedule were also completed. Results: The BWS group demonstrated higher blood-oxygen level-dependent (BOLD) signal change in the insula, visual cortex, and periaqueductal gray, whereas the CON group displayed increased hypothalamus activation in response to high amplitude negative-valence (HAN) image presentation. Psychophysiological interaction (PPI) analyses found CON showed significant interactions between frontal and temporal cortical regions as well as between the hypothalamus and the insula, frontal, and temporal cortices during HAN image presentation that were not seen in BWS. No differences between groups were found in their responses to the questionnaires. Conclusion: Our findings demonstrate significant differences in brain activation between BWS and CON in response to the presentation of HAN IAPS images, despite no significant differences in scores on psychometric questionnaires.

Voice Biofeedback via Bone Conduction Headphones: Effects on Acoustic Voice Parameters and Self-Reported Vocal Effort in Individuals With Voice Disorders.

PURPOSE: This study explores sidetone amplification (amplified playback of one's own voice) provided via bone conduction in participants with voice disorders. The effects of bone conduction feedback on acoustic voice parameters and vocal effort ratings are examined. METHODS: Speech samples of 47 participants with voice disorders were recorded in three auditory feedback conditions: two with sidetone amplification delivered via bone conduction and one condition with no alteration of the feedback. After each task, the participants rated their vocal effort on a visual analog scale. The voice recordings were evaluated by a speech-language pathologist through the GRBAS scale and processed to calculate the within-participant centered sound pressure level (SPL) values, the mean pitch strength (PS), the time dose (Dt%), and cepstral peak prominence smoothed (CPPS). The effects of the feedback conditions on these acoustic parameters and vocal effort ratings were analyzed. RESULTS: The high sidetone amplification condition resulted in a statistically significant decrease in the within-participant centered SPL values and mean pitch strength across all participants. The feedback conditions had no statistically significant effects on the vocal effort ratings, time dose (Dt%), or CPPS. CONCLUSIONS: This study provides an evidence that bone conduction sidetone amplification contributes to a consistent adaptation in the within-participant centered SPL values (ΔSPL) in patients with vocal hyperfunction, glottal insufficiency, and organic/neurological laryngeal pathologies compared to conditions with no feedback.

Evaluating the clinical benefit of brain-computer interfaces for control of a personal computer.

Brain-computer interfaces (BCIs) enabling the control of a personal computer could provide myriad benefits to individuals with disabilities including paralysis. However, to realize this potential, these BCIs must gain regulatory approval and be made clinically available beyond research participation. Therefore, a transition from engineering-oriented to clinically oriented outcome measures will be required in the evaluation of BCIs. This review examined how to assess the clinical benefit of BCIs for the control of a personal computer. We report that: (a) a variety of different patient-reported outcome measures can be used to evaluate improvements inhow a patient feels, and we offer some considerations that should guide instrument selection. (b) Activities of daily living can be assessed to demonstrate improvements inhow a patient functions, however, new instruments that are sensitive to increases in functional independence via the ability to perform digital tasks may be needed. (c) Benefits tohow a patient surviveshas not previously been evaluated but establishing patient-initiated communication channels using BCIs might facilitate quantifiable improvements in health outcomes.

Safety and Tolerability of a Wearable, Vibrotactile Stimulation Device for Parkinson's Disease.

Background: Resting tremor is a cardinal symptom of Parkinson's disease (PD) that contributes to the physical, emotional, and economic burden of the disease. Objective: The goal of this study was to investigate the safety, tolerability, and preliminary effectiveness of a novel wearable vibrotactile stimulation device on resting tremor in individuals with PD. Methods: Using a randomized cross-over design, subjects received two different vibrotactile stimulation paradigms (high amplitude patterned and low amplitude continuous) on two separate laboratory visits. On each visit, resting tremor was video recorded for 10 min at baseline and while the vibrotactile stimulation was applied. Tremor severity was scored by a blinded clinician. Results: Both vibration paradigms were well safe and well tolerated and resulted in a reduction in resting tremor severity with a moderate effect size (n = 44, p < 0.001, r = 0.37-0.54). There was no significant difference between the two vibration paradigms (p = 0.14). Conclusion: Short durations of vibrotactile stimulation delivered via wearable devices were safe and well tolerated and may attenuate resting tremor severity in individuals with PD. The sample size as well as the potential preliminary effectiveness revealed by two arms of the study could not eliminate the potential for a placebo effect.

Music Augmented With Isochronic Auditory Beats or Vibrotactile Stimulation Does Not Affect Subsequent Ergometer Cycling Performance: A Pilot Study.

Methods to enhance the ergogenic effects of music are of interest to athletes of all abilities. The aim of this pilot study was to investigate the ergogenic effects of two commercially available methods of music augmentation: auditory beats and vibrotactile stimulation. Six male and five female cyclists/triathletes cycled for 7 minutes at three different intensities: a rate of perceived exertion (RPE) of 11 ("light"), RPE of 15 ("hard"), and a 7-minute time-trial. Before each 7-minute bout of cycling, participants listened to 10 minutes of self-selected music (MUS), or the same music with the addition of either isochronic auditory beats (ABS) or vibrotactile stimulation via SUBPACTM (VIB). MUS, ABS and VIB trials were performed in a randomized order. Power output was measured during cycling and felt arousal and feeling scores were recorded at timepoints throughout the protocol. The results found the augmented MUS interventions did not influence power output with no significant main effect of trial (p = 0.44, η2 = 0.09) or trial × cycling intensity interaction (p = 0.11, η2 = 0.20). Similarly, both felt arousal and feeling scores were unchanged between the MUS, ABS, and VIB trials (p > 0.05). In conclusion, this pilot study indicated an ineffectiveness of the ABS and VIB to affect subsequent 7-min cycling performance compared to self-selected MUS alone.

Singing In The Mask: Effects Of A Variably Occluded Face Mask On Singing.

OBJECTIVE: A limitation of traditional semi-occluded vocal tract exercises (SOVTE) is the single sustained vowel task that precludes co-articulated singing. This study investigated immediate effects of a variably occluded face mask (VOFM) on vocal effort, acoustic, and aerodynamic measures in sung low and high pitches of healthy singers. DESIGN: Single-group, pre-post intervention study. METHODS: The outlet ports of disposable anesthesia facemasks were fitted with plastic caps with two drilled openings sizes (9.6 mm, 6.4 mm). Twenty-three singers with no voice complaints provided baseline vocal effort, acoustic, and aerodynamic measures in high and low pitches. Participants trained in four conditions: two VOFM sizes (9.6 mm, 6.4 mm) in combination with the 20th and 80th percentile of the singer's pitch range. Participants were trained on three phonatory tasks: repeated consonant/vowel syllables, sung sentence, and sustained vowel. Vocal effort before and after training was compared using a visual-analog scale, while standardized mean differences captured acoustic and aerodynamic changes before and after training. RESULTS: Participants reported decreased vocal effort after VOFM training at all occlusion and pitch combinations. On average, consistent beneficial effect sizes were found in cepstral peak prominence (CPP) and cepstral spectral index of dysphonia (CSID) for all 4 occlusion-pitch combinations, and vocal intensity and mean estimated subglottal pressure increased for all 4 occlusion-pitch training combinations. Changes in mean phonatory airflow and resistance were less consistent. DISCUSSION: There was an immediate effect of decreased vocal effort in singing after VOFM training. Acoustic and aerodynamic effects were variable and modest. Future studies should explore changes in these outcomes after VOFM in singing voice therapy.

Parkinson's Disease With Visual Hallucinations Is Associated With Epileptiform Activity on EEG.

Background: Visual hallucinations (VHs) in Parkinson's disease (PD) are the cardinal symptoms which declare the onset of PD psychosis (PDP). The anthropomorphic and zoomorphic VHs of PD resemble those of Charles Bonnet syndrome and temporal lobe epilepsy. In both of these disorders electroencephalography (EEG) abnormalities have been described. We therefore sought to examine whether VHs in PD were associated with similar EEG abnormalities. Methods: This retrospective observational study searched the medical records of 300 PD patients and filtered for those containing clinical 20-min scalp EEGs. Remaining records were separated into two groups: patients with reported VHs and those without. The prevalence of epileptiform discharges in the EEGs of both groups was identified. Results: Epileptiform discharges were present in 5 of 13 (38.5%) PD patients with VHs; all localized to the temporal lobe. No epileptiform discharges were observed in the EEGs of the 31 PD patients without VHs. Conclusion: The significantly high incidence of temporal lobe epileptiform discharges in PD patients with VHs as compared to those without VHs lends to the possibility of an association visual cortex epileptogenic focus. Accordingly, for treatment-refractory patients, antiepileptic drugs might be considered, as in the case of Charles Bonnet syndrome, temporal lobe epilepsy and migraine with visual aura. Future prospective studies involving larger samples and multi-center cohorts are required to validate these observational findings.

Electrocorticography reveals thalamic control of cortical dynamics following traumatic brain injury.

The return of consciousness after traumatic brain injury (TBI) is associated with restoring complex cortical dynamics; however, it is unclear what interactions govern these complex dynamics. Here, we set out to uncover the mechanism underlying the return of consciousness by measuring local field potentials (LFP) using invasive electrophysiological recordings in patients recovering from TBI. We found that injury to the thalamus, and its efferent projections, on MRI were associated with repetitive and low complexity LFP signals from a highly structured phase space, resembling a low-dimensional ring attractor. But why do thalamic injuries in TBI patients result in a cortical attractor? We built a simplified thalamocortical model, which connotes that thalamic input facilitates the formation of cortical ensembles required for the return of cognitive function and the content of consciousness. These observations collectively support the view that thalamic input to the cortex enables rich cortical dynamics associated with consciousness.

Cardiac arrest after severe traumatic brain injury can be survivable with good outcomes.

BACKGROUND: Resuscitation for traumatic cardiac arrest (TCA) in patients with severe traumatic brain injury (sTBI) has historically been considered futile. There is little information on the characteristics and outcomes of these patients to guide intervention and prognosis. The purpose of the current study is to report the clinical characteristics, survival, and long-term neurological outcomes in patients who experienced TCA after sTBI and analyze the factors contributing to survival. METHODS: A retrospective review identified 42 patients with TCA from a total of 402 patients with sTBI (Glasgow Coma Scale (GCS) score ≤8) who were admitted to Stony Brook University Hospital, a level I trauma center, from January 2011 to December 2018. Patient demographics, clinical characteristics, survival, and neurological functioning during hospitalization and at follow-up visits were collected. RESULTS: Of the 42 patients, the average age was 45 years and 21.4% were female. Eight patients survived the injury (19.0%) to discharge and seven survived with good neurological function. Admission GCS score and bilateral pupil reactivity were found to be significant indicators of survival. The mean GCS score was 5.3 in survivors and 3.2 in non-survivors (p=0.020). Age, Injury Severity Score, or cardiac rhythm was not associated with survival. Frequent neuroimaging findings included subarachnoid hemorrhage, subdural hematoma, and diffuse axonal injury. DISCUSSION: TCA after sTBI is survivable and seven out of eight patients in our study recovered with good neurological function. GCS score and pupil reactivity are the best indicators of survival. Our results suggest that due to the possibility of recovery, resuscitation and neurosurgical care should not be withheld from this patient population. LEVEL OF EVIDENCE: Level IV, therapeutic/care management.

Modeling the return to consciousness after severe traumatic brain injury at a large academic level 1 trauma center.

OBJECTIVE: Severe traumatic brain injury (sTBI) carries significant morbidity and mortality. It remains difficult to counsel families on functional prognosis and plan research initiatives aimed at treating traumatic coma. In order to better address these problems, the authors set out to develop statistical models using retrospective data to identify admission characteristics that correlate with time until the return of consciousness, defined as the time to follow commands (TFC). These results were then used to create a TFC score, allowing for rapid identification of patients with predicted prolonged TFC. METHODS: Data were reviewed and collected from medical records of sTBI patients with Glasgow Coma Scale (GCS) motor subscores ≤ 5 who were admitted to Stony Brook University Hospital from January 2011 to July 2018. Data were used to calculate descriptive statistics and build binary logistic regression models to identify admission characteristics that correlated with in-hospital mortality and in-hospital command-following. A Cox proportional hazards model was used to identify admission characteristics that correlated with the length of TFC. A TFC score was developed using the significant variables identified in the Cox regression model. RESULTS: There were 402 adult patients who met the inclusion criteria for this study. The average age was 50.5 years, and 122 (30.3%) patients were women. In-hospital mortality was associated with older age, higher Injury Severity Score (ISS), higher Rotterdam score (head CT grading system), and the presence of bilateral fixed and dilated pupils (p < 0.01). In-hospital command-following was anticorrelated with age, ISS, Rotterdam score, and the presence of a single fixed and dilated pupil (p < 0.05). TFC was anticorrelated with age, ISS, Rotterdam score, and the presence of a single fixed and dilated pupil. Additionally, patients who sustained injuries from falls from standing height had a shorter average TFC. The 3 significant variables from the Cox regression model that explained the most variance were used to create a 4-point TFC score. The most significant of these characteristics were Rotterdam head CT scores, high impact traumas, and the presence of a single fixed and dilated pupil. Importantly, the presence of a single fixed and dilated pupil was correlated with longer TFC but no increase in likelihood of in-hospital mortality. CONCLUSIONS: The creation of the 4-point TFC score will allow clinicians to quickly identify patients with predicted prolonged TFC and estimate the likelihood of command-following at different times after injury. Discussions with family members should take into account the likelihood that patients will return to consciousness and survive after TBI.

An automated, electronic assessment tool can accurately classify older adult postural stability.

Current methods of balance assessment in the clinical environment are often subjective, time-consuming and lack clinical relevance for non-ambulatory older adults. The objective of this study was to develop a novel method of balance assessment that utilizes data collected using the Microsoft Kinect 2 to create a Berg Balance Scale score, which is completely determined by statistical methods rather than by human evaluators. 74 older adults, both healthy and balance impaired, were recruited for this trial. All participants completed the Berg Balance Scale (BBS) which was scored independently by trained physical therapists. Participants then completed the items of the "Modified Berg Balance Scale" in front of the Microsoft Kinect camera. Kinematic data collected during this measurement was used to train a feed-forward neural network that was used to assign a Berg Balance Scale score. The neural network model estimated the clinician-assigned BBS score to within a median of 0.93 points for the participants in our sample population (range: 0.02-5.69). Using low-cost depth sensing camera technology and a clinical protocol that takes less than 5 min to complete in both ambulatory and non-ambulatory older adults, the method outlined in this manuscript can accurately predict a participant's BBS score and thereby identify whether they are deemed a high fall risk or not. If implemented correctly, this could enable fall prevention services to be deployed in a timely fashion using low-cost, accessible technology, resulting in improved safety of older adults.

The tripartite associations between bacteriophage, Wolbachia, and arthropods.

By manipulating arthropod reproduction worldwide, the heritable endosymbiont Wolbachia has spread to pandemic levels. Little is known about the microbial basis of cytoplasmic incompatibility (CI) except that bacterial densities and percentages of infected sperm cysts associate with incompatibility strength. The recent discovery of a temperate bacteriophage (WO-B) of Wolbachia containing ankyrin-encoding genes and virulence factors has led to intensifying debate that bacteriophage WO-B induces CI. However, current hypotheses have not considered the separate roles that lytic and lysogenic phage might have on bacterial fitness and phenotype. Here we describe a set of quantitative approaches to characterize phage densities and its associations with bacterial densities and CI. We enumerated genome copy number of phage WO-B and Wolbachia and CI penetrance in supergroup A- and B-infected males of the parasitoid wasp Nasonia vitripennis. We report several findings: (1) variability in CI strength for A-infected males is positively associated with bacterial densities, as expected under the bacterial density model of CI, (2) phage and bacterial densities have a significant inverse association, as expected for an active lytic infection, and (3) CI strength and phage densities are inversely related in A-infected males; similarly, males expressing incomplete CI have significantly higher phage densities than males expressing complete CI. Ultrastructural analyses indicate that approximately 12% of the A Wolbachia have phage particles, and aggregations of these particles can putatively occur outside the Wolbachia cell. Physical interactions were observed between approximately 16% of the Wolbachia cells and spermatid tails. The results support a low to moderate frequency of lytic development in Wolbachia and an overall negative density relationship between bacteriophage and Wolbachia. The findings motivate a novel phage density model of CI in which lytic phage repress Wolbachia densities and therefore reproductive parasitism. We conclude that phage, Wolbachia, and arthropods form a tripartite symbiotic association in which all three are integral to understanding the biology of this widespread endosymbiosis. Clarifying the roles of lytic and lysogenic phage development in Wolbachia biology will effectively structure inquiries into this research topic.

Robotic Rehabilitation and Spinal Cord Injury: a Narrative Review.

Mobility after spinal cord injury (SCI) is among the top goals of recovery and improvement in quality of life. Those with tetraplegia rank hand function as the most important area of recovery in their lives, and those with paraplegia, walking. Without hand function, emphasis in rehabilitation is placed on accessing one's environment through technology. However, there is still much reliance on caretakers for many activities of daily living. For those with paraplegia, if incomplete, orthoses exist to augment walking function, but they require a significant amount of baseline strength and significant energy expenditure to use. Options for those with motor complete paraplegia have traditionally been limited to the wheelchair. While wheelchairs provide a modified level of independence, wheelchair users continue to face difficulties in access and mobility. In the past decade, research in SCI rehabilitation has expanded to include external motorized or robotic devices that initiate or augment movement. These robotic devices are used with 2 goals: to enhance recovery through repetitive, functional movement and increased neural plasticity and to act as a mobility aid beyond orthoses and wheelchairs. In addition, lower extremity exoskeletons have been shown to provide benefits to the secondary medical conditions after SCI such as pain, spasticity, decreased bone density, and neurogenic bowel. In this review, we discuss advances in robot-guided rehabilitation after SCI for the upper and lower extremities, as well as potential adjuncts to robotics.

Nuclear-mitochondrial epistasis and drosophila aging: introgression of Drosophila simulans mtDNA modifies longevity in D. melanogaster nuclear backgrounds.

Under the mitochondrial theory of aging, physiological decline with age results from the accumulated cellular damage produced by reactive oxygen species generated during electron transport in the mitochondrion. A large body of literature has documented age-specific declines in mitochondrial function that are consistent with this theory, but relatively few studies have been able to distinguish cause from consequence in the association between mitochondrial function and aging. Since mitochondrial function is jointly encoded by mitochondrial (mtDNA) and nuclear genes, the mitochondrial genetics of aging should be controlled by variation in (1) mtDNA, (2) nuclear genes, or (3) nuclear-mtDNA interactions. The goal of this study was to assess the relative contributions of these factors in causing variation in Drosophila longevity. We compared strains of flies carrying mtDNAs with varying levels of divergence: two strains from Zimbabwe (<20 bp substitutions between mtDNAs), strains from Crete and the United States (approximately 20-40 bp substitutions between mtDNAs), and introgression strains of Drosophila melanogaster carrying mtDNA from Drosophila simulans in a D. melanogaster Oregon-R chromosomal background (>500 silent and 80 amino acid substitutions between these mtDNAs). Longevity was studied in reciprocal cross genotypes between pairs of these strains to test for cytoplasmic (mtDNA) factors affecting aging. The intrapopulation crosses between Zimbabwe strains show no difference in longevity between mtDNAs; the interpopulation crosses between Crete and the United States show subtle but significant differences in longevity; and the interspecific introgression lines showed very significant differences between mtDNAs. However, the genotypes carrying the D. simulans mtDNA were not consistently short-lived, as might be predicted from the disruption of nuclear-mitochondrial coadaptation. Rather, the interspecific mtDNA strains showed a wide range of variation that flanked the longevities seen between intraspecific mtDNAs, resulting in very significant nuclear x mtDNA epistatic interaction effects. These results suggest that even "defective" mtDNA haplotypes could extend longevity in different nuclear allelic backgrounds, which could account for the variable effects attributable to mtDNA haplogroups in human aging.

The roles of positive and negative selection in the molecular evolution of insect endosymbionts.

The evolutionary rate acceleration observed in most endosymbiotic bacteria may be explained by higher mutation rates, changes in selective pressure, and increased fixation of deleterious mutations by genetic drift. Here, we explore the forces influencing molecular evolution in Blochmannia, an obligate endosymbiont of Camponotus and related ant genera. Our goals were to compare rates of sequence evolution in Blochmannia with related bacteria, to explore variation in the strength and efficacy of negative (purifying) selection, and to evaluate the effect of positive selection. For six Blochmannia pairs, plus Buchnera and related enterobacteria, estimates of sequence divergence at four genes confirm faster rates of synonymous evolution in the ant mutualist. This conclusion is based on higher dS between Blochmannia lineages despite their more recent divergence. Likewise, generally higher dN in Blochmannia indicates faster rates of nonsynonymous substitution in this group. One exception is the groEL gene, for which lower dN and dN/dS compared to Buchnera indicate exceptionally strong negative selection in Blochmannia. In addition, we explored evidence for positive selection in Blochmannia using both site-and lineage-based maximum likelihood models. These approaches confirmed heterogeneity of dN/dS among codon sites and revealed significant variation in dN/dS across Blochmannia lineages for three genes. Lineage variation affected genes independently, with no evidence of parallel changes in dN/dS across genes along a given branch. Our data also reveal instances of dN/dS greater than one; however, we do not interpret these large dN/dS ratios as evidence for positive selection. In sum, while drift may contribute to an overall rate acceleration at nonsynonymous sites in Blochmannia, variable selective pressures best explain the apparent gene-specific changes in dN/dS across lineages of this ant mutualist. In the course of this study, we reanalyzed variation at Buchnera groEL and found no evidence of positive selection that was previously reported.

Wolbachia interactions that determine Drosophila melanogaster survival.

We have recently described a mutualistic symbiosis in which Wolbachia bacteria were shown to improve the fitness of some Drosophila melanogaster stocks. Wolbachia did not extend longevity in all Drosophila genotypes, even though 16s rDNA sequences indicated that our Drosophila stocks were infected with the same Wolbachia strain. Here, we use reciprocal hybrid crosses between two Drosophila strains, one that lived longer with Wolbachia (Z53) and one that did not (Z2), to investigate the inheritance of the survival phenotype and its dependence on the host genotype, sex, and mating conditions. Wolbachia's positive effects were more apparent in hybrid flies than in parental flies, ruling out exclusive maternal inheritance or the dependence of the survival phenotype on Wolbachia strain differences. The Wolbachia survival effects were more apparent in single-sex cages, where courtship and mating were not permitted. In these cages, nearly all flies with Wolbachia lived longer than uninfected flies, even though strain Z2 showed no Wolbachia effect in mixed-sex mating cages. We used comparisons between single- and mixed-sex cages to estimate the cost of reproduction for both sexes. Our data suggest that Wolbachia infection may increase the inferred cost of reproduction, particularly in males. Wolbachia can even produce a positive survival effect almost as large as the negative survival effect associated with reproduction. We discuss the implications of our experiments for the study of insect symbioses.