Iain Anthony

The Neuropathology of HIV/AIDS

The introduction of Highly Active Anti-retroviral Therapy (HAART) has resulted in significant decreases in morbidity and mortality for subjects infected with HIV. The brain is a major target organ for HIV resulting in significant neuropathological changes in most HIV infected subjects and a wide range of clinical neurological symptoms including HIV associated dementia. In the pre-HAART era HIV associated dementia was a common complication of AIDS. However, since the introduction of HAART the incidence of HIV associated dementia has fallen, but the prevelance has actually risen due to the increasing number of infected subjects and increased life expectancy. HIV associated dementia correlates most closely with neuroinflammation rather than directly with viral load or HIV encephalitis. HIV related clinical and neuropathological disorders are more prevalent in drug abusers than in other risk groups. This review focuses on the shifting pathology observed in HIV infected subjects since the introduction of HAART, discussing the clinical manifestations of these and the influence of confounding factors such as drug abuse and Hepatitis C co-infection.

Reactivation and Mutation of Newly Discovered WU, KI, and Merkel Cell Carcinoma Polyomaviruses in Immunosuppressed Individuals

BACKGROUND Infection with the human polyomaviruses BK (BKV) and JC (JCV) is almost ubiquitous, asymptomatic, and lifelong. However, reactivation during immunosuppression, associated with mutations in the transcriptional control region (TCR) that up-regulates viral replication, can cause life-threatening disease. In this study, we investigated whether the recently discovered WU and KI polyomaviruses (WUPyV and KIPyV) and Merkel cell polyomavirus (MCPyV) could, like BKV and JCV, persist, mutate, and reactivate in immunodeficient subjects. METHODS Autopsy samples of lymphoid tissue from 42 AIDS-immunosuppressed subjects and 55 control samples were screened by polymerase chain reaction for all 5 polyomaviruses. TCR sequences from KIPyV and WUPyV recovered from both immunosuppressed and nonimmunosuppressed subjects were compared. RESULTS Combined polyomavirus detection frequencies were much higher for the immunosuppressed group, compared with the nonimmunosuppressed group (35.7% vs. 3.6%), with viral loads in lymphoid tissues ranging from < or = 8.4 x 10(5) to > 1.5 x 10(5) viral genome copies per 10(6) cells. MCPyV was recovered from only 1 HIV-negative study subject. TCR sequences from reactivated WUPyV and KIPyV variants showed a number of point mutations and insertions that were absent in viruses recovered from respiratory tract specimens obtained from nonimmunosuppressed subjects. CONCLUSIONS KIPyV and WUPyV show reactivation frequencies comparable to those of BKV and JCV during immunosuppression. TCR changes that potentially lead to transcriptional dysregulation may have pathogenic consequences equivalent in severity to those observed for JCV and BKV.

Accelerated Tau deposition in the brains of individuals infected with human immunodeficiency virus-1 before and after the advent of highly active anti-retroviral therapy

This study aims to investigate the influence of human immunodeficiency virus (HIV) infection on the neurodegenerative processes normally associated with ageing. We have looked for evidence of beta amyloid and hyperphosphorylated Tau deposition in HIV-infected subjects before and after the advent of highly active anti-retroviral therapy (HAART). In addition we have looked for evidence of axonal damage. We have compared these HIV-positive cases with age-matched controls and with older non-demented controls. We find no evidence of significant premature beta amyloid deposition in HIV-infected cases; however, we do observe elevated levels of hyperphosphorylated Tau in the hippocampus of many HIV-infected subjects, compared with age-matched controls. The greatest levels of hyperphosphorylated Tau are noted in HAART-treated subjects. Axonal damage marked by expression of beta amyloid pre-cursor protein (BAPP) was highly variable in all groups including control subjects. We surmise that HIV infection and/or the use of anti-retroviral therapy may predispose to accelerated neuroageing in the form of hyperphosphorylated Tau deposition in the hippocampus. Within the age groups studied these significant neuropathological changes remained subclinical and were not yet associated with cognitive impairment.

Neurobiology of multiple insults: HIV-1-associated brain disorders in those who use illicit drugs.

Despite two decades of research, certain aspects of HIV-related central nervous system (CNS) disease remain poorly understood. HIV targets microglia and macrophages within the CNS and enters the brain compartment early. However, HIV is there held in check apparently until the onset of significant immune compromise, when viral replication, microglial activation, neuronal damage, and cognitive impairment are likely to ensue. Illicit drug abuse continues to be a significant risk factor for HIV transmission worldwide. Whether HIV-related CNS disease is more prevalent or more severe in this risk group has long been debated. Drugs of abuse can of themselves cause immune suppression, blood–brain barrier breakdown, microglial activation, and neuronal injury. This review presents evidence that HIV associated CNS disorders are indeed accentuated in drug abusers. However, the advent of effective therapy has added a new dimension, which must be taken into consideration. Treated individuals are surviving much longer and HIV encephalitis and HIV-associated dementia have become much less common. However, more subtle forms of CNS damage are emerging. Examination of the brains of individuals who have been treated long term with highly active antiretroviral therapy (HAART) reveals a surprising degree of microglial activation, comparable at times to that seen formerly in milder cases of HIV encephalitis. In addition, these individuals show evidence of increased deposition of neurodegenerative proteins, particularly hyperphosphorylated tau. Similar observations have been made in young opiate abusers who are HIV negative. Taken together, these results suggest that neuroinflammation and neurodegeneration, which are clinically silent at present, may cause problems in the future in HAART-treated subjects.

Component rotational alignment in unexplained painful primary total knee arthroplasty.

BACKGROUND Rotational malalignment of the components in total knee arthroplasty (TKA) can be a factor in poor outcomes but has yet to be defined. This study compares the rotational alignment of components in a cohort of 56 patients with unexplained pain following total knee arthroplasty with a matched control cohort of 56 patients with cemented Nex Gen Legacy posterior stabilised (LPS) flex fixed bearing TKA between March 2006 and May 2010. The aim of the study was to define an acceptable limit of rotation in total knee replacement. METHODS Rotational alignment was calculated using the Berger protocol with post operative computerised tomography scanning. The alignment parameters measured were tibial and femoral component rotations and the combined component rotations and the component rotational mismatch. RESULTS The two cohorts were demographically matched. Excessive internal rotation of the components was defined using the tenth percentile of rotations in the control cohort. Values of excessive internal rotation were 5.8° of the tibial component, 3.9° of the femoral component, 8.7° of combined rotation and 5.6° of the component mismatch. No significant difference was identified in excessive external rotation in any of the parameters. A significant difference in the mean rotations between the two cohorts was identified with internal rotation of the components in the painful cohort and external rotation on the control cohort. CONCLUSIONS We identified internal rotation malalignment of the tibial (p=0.0003) and femoral (p=0.014) components individually as well as the combined component rotation (p=0.0003) and component rotation mismatch (p=0.0001) to be a factor in pain following TKA. External rotation of any of the component parameters was not identified to be a factor in painful TKA. This study adds to the understanding of rotational alignment in TKA and suggests limits of internal rotation alignment associated with painful Nex Gen Legacy posterior stabilised (LPS) flex fixed bearing TKA. LEVEL OF EVIDENCE Level III.

Predisposition to accelerated Alzheimer-related changes in the brains of human immunodeficiency virus negative opiate abusers

Cognitive impairment is a recognized effect of drug misuse, including the use of opiates. The pathological basis for this is unknown but the temporal and frontal cortices have been implicated. We have shown previously that deposits of hyperphosphorylated tau in drug user brains exceed those seen in age-matched controls. The present quantitative study of hyperphosphorylated tau and beta amyloid in drug user brains allows comparison with the related pathology in Alzheimers disease. Brains were obtained from the Edinburgh Medical Research Council Brain Banks, comprising 39 human immunodeficiency virus negative drug users, five subjects with Alzheimers disease and 37 age-matched, cognitively normal controls, all legally and ethically approved for research. Hyperphosphorylated tau positive (AT8, AT100) neuropil threads were significantly increased in the frontal and temporal cortex, and in the locus coeruleus, of drug users aged > 30 years (all P = 0.04). Under the age of 30 years, drug users showed a similar increase in neuropil threads compared with controls, but this reached significance only in the frontal cortex (P = 0.03). Immunopositivity for both three- and four-repeat tau was present in drug user brains. There was a direct relationship between the numbers of neuropil threads and of neurofibrillary tangles: neurofibrillary tangles were sparse in brains that had neuropil thread counts below 200 cm(2). Hyperphosphorylated tau positive neuropil threads increased at a faster rate in drug users than in controls and the levels of the phosphorylating enzyme, GSK-3, was raised in drug user brains. Beta amyloid (AB4, AB42 and 4G8) was raised in drug user brains (mainly as shadow plaques) but not significantly different from controls and there was no correlation between high beta amyloid and hyperphosphorylated tau in individual cases. Hyperphosphorylated tau levels correlated significantly (P = 0.038) with microglial activation in drug users but not in controls. The levels of hyperphosphorylated tau in drug users fell far short of those seen in Alzheimers disease but overlapped with those in elderly controls. We conclude that drug users show early Alzheimers disease-related brain pathology that may be the basis for cognitive impairment and that neuroinflammation is an early accompanying feature. This provides an opportunity to study the pathogenesis of tau pathology in the human brain.

Hyperphosphorylated tau and amyloid precursor protein deposition is increased in the brains of young drug abusers

Drug abuse is a major problem worldwide. The incidence of drug‐related deaths attributed to opiate abuse is increasing annually. Apart from routine examination, little is known of the neuropathology of drug abuse. We, and others, have shown previously that drug abuse is associated with microglial activation. We hypothesised that neuroinflammation might lead to premature neurodegeneration in drug abusers. We investigated the brains of young opiate abusers (n = 34, all < 40 years) for the presence of proteins associated with neurodegenerative diseases and compared them with the brains of age‐matched, non‐drug users (n = 16) all of whom died suddenly. Detailed immunohistochemical analysis of the hippocampus, brainstem and basal ganglia for hyperphosphorylated tau, β‐amyloid, β‐amyloid precursor protein (βAPP) and ubiquitin demonstrated an excess of AT8‐positive neurofibrillary tangles (NFT) in the drug abusers. These were not only more prevalent in the drug abusers than in controls (44%vs. 19%) but also involved more brain areas. In controls NFT were confined to the entorhinal cortex whereas in drug users they were also found in the subiculum, temporal neocortex, nucleus basalis of Meynert and the locus coeruleus. Virtually no amyloid plaques were present but βAPP positivity was again much more common in drug abusers than controls (73%vs. 20% in the brainstem and 59%vs. 23% in the temporal lobe). There is no suggestion that these drug abusers had displayed major cognitive impairment although detailed neuropsychological assessment is difficult in this subject group. Likely causes of hyperphosphorylated tau deposition in drug abuse include hypoxic‐ischaemic injury, microglial‐associated cytokine release and possibly drug‐associated neurotoxicity or hepatitis. Head injury, which is another major risk factor, does not appear to have contributed to our findings. Genetic factors also merit consideration. It is unclear at present how much of the hyperphosphorylated tau detected in these young drug abusers represents a transitory phenomenon.

The effects of illicit drugs on the HIV infected brain.

Evidence accumulating from clinical observations, neuroimaging and neuropathological studies suggests that illicit drug abuse accentuates the adverse effects of HIV on the central nervous system (CNS). Experimental investigation in cell culture models supports this conclusion. Injecting drug abuse is also a risk factor for the acquisition of HIV infection, the incidence of which continues to rise in intravenous drug users (IVDU) even in countries with access to effective therapy. In order to understand the interactions of drug abuse and HIV infection, it is necessary to examine the effects of each insult in isolation before looking for their combined effects. This review traces progress in understanding the pathogenesis of HIV related CNS disorders before the introduction of effective therapy and compares the state of our knowledge now that effective therapy has significantly modified disease progression. The additional impact of intravenous drug abuse on HIV-associated brain disease, then and now, is also reviewed. Predictions for the future are discussed, based on what is known at present and on recently emerging data.

Does drug abuse alter microglial phenotype and cell turnover in the context of advancing HIV infection

The aim of this study was to test the effects of drug abuse, in particular opiate abuse, on the phenotype and turnover of microglial cells within the brain in the context of advancing HIV infection. Basal ganglia and hippocampus sections were studied in 51 cases divided into six groups: HIV‐negative normal controls, HIV‐negative drug abusers, AIDS nondrug abusers, AIDS drug abusers, HIV encephalitis (HIVE) nondrug abusers and HIVE drug abusers. None of the cases studied had received highly active anti‐retroviral therapy (HAART). Microglial phenotypes were defined using CD14, CD16, CD68 and major histocompatibility class II (MHC II). Microglial turnover was assessed using terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) (DNA damage), BAX (proapoptotic marker), Fas (CD95) (proapoptotic), proliferating cell nuclear antigen (PCNA) (proliferation and DNA repair), Ki‐67 (cell proliferation) and BCL‐2 (antiapoptosis). We find increased expression of MHC II and CD16 in response to drug abuse. We also noted increased levels of TUNEL positivity in drug abusers compared to nondrug abusers, although conversely we found lower levels of BAX in those who had abused drugs. We find no evidence of microglial proliferation in any of our study groups, including HIVE, although HIV infection leads to increased expression of CD16, CD68 and MHC II. CD14 expression was restricted to perivascular microglia in all groups (including normal controls) apart from the two HIVE groups where some but not all cases also showed parenchymal expression of CD14. In contrast, CD16 was found in parenchymal microglia in all groups. Using high‐pressure antigen retrieval and tyramide signal amplification, we find moderately high levels of CD16 expression in the parenchyma of normal brains which is not normally observed using standard avidin/biotin complex (ABC) techniques. This suggests that a low basal expression of CD16 occurs in many resident microglial cells which may potentially be upregulated in HIV‐infected individuals. From these data, we suggest that not all the CD16+ parenchymal cells detected in AIDS brains (using ABC) represent influx of monocyte lineage cells from the blood. Finally the increased expression of MHC II and CD68 detected in drug abusers with HIVE compared to nondrug abusers with HIVE suggests that the combination of drug abuse and HIV infection has a greater deleterious effect on the brain than either individual insult on its own.

Improved Accuracy of Component Positioning with Robotic-Assisted Unicompartmental Knee Arthroplasty: Data from a Prospective, Randomized Controlled Study.

BACKGROUND Higher revision rates have been reported in patients who have undergone unicompartmental knee arthroplasty compared with patients who have undergone total knee arthroplasty, with poor component positioning identified as a factor in implant failure. A robotic-assisted surgical procedure has been proposed as a method of improving the accuracy of component implantation in arthroplasty. The aim of this prospective, randomized, single-blinded, controlled trial was to evaluate the accuracy of component positioning in unicompartmental knee arthroplasty comparing robotic-assisted and conventional implantation techniques. METHODS One hundred and thirty-nine patients were randomly assigned to treatment with either a robotic-assisted surgical procedure using the MAKO Robotic Interactive Orthopaedic Arm (RIO) system or a conventional surgical procedure using the Oxford Phase-3 unicompartmental knee replacement with traditional instrumentation. A postoperative computed tomographic scan was performed at three months to assess the accuracy of the axial, coronal, and sagittal component positioning. RESULTS Data were available for 120 patients, sixty-two who had undergone robotic-assisted unicompartmental knee arthroplasty and fifty-eight who had undergone conventional unicompartmental knee arthroplasty. Intraobserver agreement was good for all measured component parameters. The accuracy of component positioning was improved with the use of the robotic-assisted surgical procedure, with lower root mean square errors and significantly lower median errors in all component parameters (p < 0.01). The proportion of patients with component implantation within 2° of the target position was significantly greater in the group who underwent robotic-assisted unicompartmental knee arthroplasty compared with the group who underwent conventional unicompartmental knee arthroscopy with regard to the femoral component sagittal position (57% compared with 26%, p = 0.0008), femoral component coronal position (70% compared with 28%, p = 0.0001), femoral component axial position (53% compared with 31%, p = 0.0163), tibial component sagittal position (80% compared with 22%, p = 0.0001), and tibial component axial position (48% compared with 19%, p = 0.0009). CONCLUSIONS Robotic-assisted surgical procedures with the use of the MAKO RIO lead to improved accuracy of implant positioning compared with conventional unicompartmental knee arthroplasty surgical techniques. LEVEL OF EVIDENCE Therapeutic Level I. See Instructions for Authors for a complete description of levels of evidence.