Botulinum Toxin A Treatment in HIV Infected Patients-A Long-Term Observational Study.

OBJECTIVE: It is unknown whether interactions between HIV infection and the safety of botulinum toxin A (BTX) exist. METHODS: We studied eight patients with HIV infection who were treated with BTX every three months for up to nine years. All patients were on antiretroviral treatment. The efficacy and safety of BTX were evaluated. RESULTS: Indications for BTX treatment (including off-label use), dosage of BTX, and frequency of application did not differ as compared to non-HIV infected patients. BTX treatment was effective in all HIV infected patients during a long-term observation period without loss of efficacy and without clinically relevant side effects. Only one of the eight patients showed mild side effects due to BTX, and no clinical signs of antibody development were noted. We also observed no signs of interaction with antiretroviral treatment. CD4+ cell count and viral load remained stable during the observation period. CONCLUSIONS: We conclude that BTX treatment is safe and effective in the treatment of HIV infected patients who suffer also from a condition which can be treated by BTX. It is a therapeutic option in addition to oral medication for HIV infected patients.

ABO-Incompatible Living Donor Kidney Transplantation in a Human Immunodeficiency Virus-Positive Recipient From a Human Immunodeficiency Virus-Positive Donor: A Case Report.

A human immunodeficiency virus (HIV) infection is no longer an absolute contraindication for solid organ transplantation, yet such a setting is still challenging and little explored because of general reservations and medical difficulties. We describe a 51-year-old man with end-stage renal failure due to polycystic kidney disease who underwent an ABO-incompatible kidney transplantation from his 49-year-old male partner. Early postoperative course revealed an episode of suspected acute rejection, which was successfully managed with a steroid pulse. Both donor and recipient continued to have an undetectable viral load after adjusting antiretroviral medication to renal function. To the best of our knowledge, this is the first report of a successful ABO-incompatible living donor kidney transplantation from an HIV-positive donor in an HIV-positive recipient, and this case seems to be a valuable approach with favorable results.

Storage and Transportation of "HIV" RNA in Plasma Samples up to 45°C in a Lyophilized Stabilizer.

BACKGROUND: Blood or plasma samples from rural areas are often transported under suboptimal conditions to central laboratories. The negative influence of different storage temperatures during transportation as well as long transportation times on the stability of unprotected HIV RNA is well known. Therefore, the correct and reliable quantification of HIV RNA might be very difficult. A stabilization solution for the storage and transportation of plasma samples was developed which stabilizes RNA for seven days up to 45°C without viral load changes. METHODS: Blood samples from HIV positive individuals were collected into EDTA containing tubes. The isolated plasma samples in Germany were pipetted into pre-prepared RNA stabilization tubes and incubated for seven days at 45°C. HIV-1 RNA quantification was performed on a HIV-1 LCx m 2000 system from Abbott and a Qiagen/Artus HI Virus-1 RG RT-PCR Kit on a Rotor-Gene Q PCR machine. In addition, plasma samples were collected and tested using existing SOP for storage and transportation in Nigeria. Plasma samples were treated with and without stabilization solution and the AMPLICOR HIV-1 MONITORTM test was used to determine viral load. RESULTS: Seventy-four stabilized plasma samples were tested in Germany and results were compared to those tested unprotected within two hours. No significant changes of viral load were detected up to seven days and 45°C in case of stabilized samples. In contrast RNA of the same unprotected samples was no longer detectable after one day at 45°C. Additionally, 22 plasma samples were investigated on day zero and under field conditions in Nigeria without changes of the viral load after seven days under given temperature conditions. CONCLUSIONS: No cooling chain is necessary for the storage and/or transportation of plasma samples treated with the new RNA stabilization solution for up to seven days. The use of this solution to preserve plasma RNA will be very helpful in countries where the environmental temperature is higher than 30°C, thus addressing the problem of unreliable viral load results due to suboptimal storage or transportation conditions. Further, the costs of storage and transportation of samples for viral load quantification could be significantly reduced.

Control beliefs and health locus of control in Ugandan, German and migrated sub-Saharan African HIV infected individuals.

OBJECTIVES: Little is known about the influence of control beliefs on antiretroviral drug adherence in patients who migrated from sub-Saharan Africa to Europe. The aim of this study was to explore the differences in health locus of control and control beliefs between HIV infected patients from sub-Saharan Africa with and without a lifetime experience of migration. METHODS: A sample of 62 HIV infected consecutive patients referred to the HIV clinics at the University Hospital of Münster (Germany) and at the Rubaga Hospital Kampala (Uganda) were enrolled into this study. We compared three groups of patients: sub-Saharan African migrants, German patients, and local Ugandan patients. We used the German health and illness related control beliefs questionnaire (KKG), the Competence and control beliefs questionnaire (FKK), and the Powe Fatalism Inventory-HIV/AIDS-Version (PFI-HIV/AIDS-Version) and translated these scales into English and Luganda. In addition, the patients' sociodemographic, acculturation, clinical, and immunological data were registered. RESULTS: Significant results were shown in HIV related external locus of control between migrated sub-Saharan African and local Ugandan patients compared to German patients. General control beliefs showed no significant differences. In the PFI-HIV-Version, there was a significant difference between migrated sub-Saharan African and Ugandan patients compared to German patients. CONCLUSIONS: Our data suggest that the experience of migration does not influence the locus of control. Compared to German HIV patients, African patients in general showed a significantly higher external health locus of control which might have implications for drug adherence.

Neurocognitive decline in HIV patients is associated with ongoing T-cell activation in the cerebrospinal fluid.

OBJECTIVE: HIV-associated neurocognitive disorders (HAND) remain a challenge despite combination antiretroviral therapy (cART). Immune cell activation has been implicated to play a major role in the development of HAND. METHODS: In this study, we used multicolor flow cytometry on peripheral blood (PB) and cerebrospinal fluid (CSF) samples to determine the expression of HLA-DR and programmed death-1 (PD-1) on CD4+ and CD8+ T cells in patients with chronic HIV infection. Expression levels were correlated with HI virus load in PB and CSF, classification of HAND and severity of magnetic resonance imaging (MRI) signal abnormalities. RESULTS: In a cohort of 86 HIV patients we found that the grade of neurocognitive impairment and the severity of MRI signal abnormalities correlated with decreasing CD4/CD8-ratios and increased frequencies of HLA-DR expressing CD4+ and CD8+ T cells reaching the highest values in the CSF samples. Importantly, HLA-DR upregulation was still detectable in virologically suppressed HIV patients. Further, T-cell subpopulation analysis of 40 HIV patients showed a significant shift from naïve to effector memory (EM) T cells that was negatively correlated with the grade of neurocognitive impairment in the PB samples. Moreover, PD-1 was significantly increased on CD4+ memory T cells with highest levels on EM T cells in HIV patients with mild or severe neurocognitive alterations. INTERPRETATION: The CD4/CD8 ratio, the proportion of EM to naïve T cells and the immune activation profile of CD4+ and CD8+ T cells in PB and CSF might be useful parameters to monitor the efficacy of cART and to identify HIV patients at risk of further neurocognitive deterioration.

Capsaicin 8 % as a cutaneous patch (Qutenza™): analgesic effect on patients with peripheral neuropathic pain.

Evaluation of the analgesic effect after a single application of the capsaicin 8 % cutaneous patch (Qutenza™) in 37 patients suffering from painful, distal symmetric polyneuropathy (PNP) for an average of 5 years. Patients ranged from 40 to 78 years of age and 22 subjects were HIV-positive. Patients were observed 4 weeks prior to 12 weeks post administration. An evaluation of the therapeutic effect of capsaicin 8 % as a dermal patch in terms of pain reduction, change of sleeping behavior and social activities was performed and statistical analysis of data was conducted using non-parametric methods. Patients were selected according to clinical criteria. Numerical rating scale (NRS 0-10) was used to inquire pain intensity and a pain score was calculated using the painDETECT(©) questionnaire Freynhagen R (Curr Med Res Opin 22:1911-1920, [2006]). A significant reduction of pain was achieved for up to 12 weeks, with a maximum after 2-4 weeks post administration. After patient education and before application of capsaicin patch, a significant reduction of three levels on the NRS was observed. Symptoms of painful PNP decreased over the period of investigation and 8 patients reported a reduction of systemic pain medication. In patients with an HIV infection, a significant extension of sleep was achieved for 2, 4 and 8 weeks after application. Thus, the application of the capsaicin 8 % patch resulted in a significant relief of neuropathic pain, a prolongation of sleep, a reduction of oral pain medication and a resumption of social activities.

Cerebrovascular events in HIV-infected patients: an analysis of a cohort of 3203 HIV+ patients during the times of cART.

PURPOSE/AIM: Cerebrovascular events (CVE) in HIV infected patients have become an increasingly relevant neurological complication. Data about the prevalence and clinical features of CVE in HIV infected patients since the introduction of combined Anti-Retroviral Therapy (cART) are rare. METHODS: A retrospective study of HIV-infected patients with a CVE was performed from 2002 to 2011. During this time period 3203 HIV-infected patients were admitted to the University hospital of Münster, Germany. All patients had access to regular and long term treatment with cART. The clinical features were analyzed and the prevalence of ischemic stroke (IS), transient ischemic attack (TIA) and intracerebral bleeding (ICB) was calculated. RESULTS: The total prevalence of all CVE was at 0.6% (95% CI: 0.3, 0.8) (0.4% for IS (95% CI: 0.2, 0.6), 0.2% for TIA (95% CI: 0.0, 0.3) and 0.1% for ICB (95% CI: 0.0, 0.2)) and the crude annual incidence rate at 59 per 100.000 for all events. The median CD4 cell count was 405/µl (25th to 75th percentile: 251-568). The majority of patients had AIDS. The median age was at 49 years (25th to 75th percentile: 40-69). Some events were associated with HIV-associated vasculopathy or viral co-infections. Most patients presented with multiple vascular risk factors. CONCLUSION: The study confirms that CVE occur in HIV-infected patients with a good immune status and at a young age. HIV infection has to be considered in young stroke patients. The rate of CVE in this study was constant when comparing to the pre-cART era. HIV associated vasculopathy and viral co-infections need to be considered in the diagnostics of stroke.

CSF penetration by antiretroviral drugs.

Severe HIV-associated neurocognitive disorders (HAND), such as HIV-associated dementia, and opportunistic CNS infections are now rare complications of HIV infection due to comprehensive highly active antiretroviral therapy (HAART). By contrast, mild to moderate neurocognitive disorders remain prevalent, despite good viral control in peripheral compartments. HIV infection seems to provoke chronic CNS injury that may evade systemic HAART. Penetration of antiretroviral drugs across the blood-brain barrier might be crucial for the treatment of HAND. This review identifies and evaluates the available clinical evidence on CSF penetration properties of antiretroviral drugs, addressing methodological issues and discussing the clinical relevance of drug concentration assessment. Although a substantial number of studies examined CSF concentrations of antiretroviral drugs, there is a need for adequate, well designed trials to provide more valid drug distribution profiles. Neuropsychological benefits and neurotoxicity of potentially CNS-active drugs require further investigation before penetration characteristics will regularly influence therapeutic strategies and outcome.

Sleepiness and sleep quality in patients with HIV infection.

OBJECTIVES: Patients with HIV infection frequently complain of sleep disturbances and daytime sleepiness. Only few data on these problems evaluated by standardized measures is available. METHODS: A sample of 180 consecutive patients with HIV infection referred to the internal and to the neurological HIV clinics at the University of Münster was enrolled in this study. The data were compared to a sample of 120 age- and sex-matched control subjects. We used the Epworth Sleepiness Scale (ESS), the Pittsburgh Sleep Quality Index (PSQI), and the Beck's Depression Inventory (BDI). In addition, the clinical and immunological data of the patients were registered. RESULTS: All scores of the ESS, the PSQI, and the BDI were significantly increased in the HIV infected patients as compared to the control group. There were no significant correlations between any of the immune parameters and the scores. Only a higher BDI score was correlated with both the ESS score and the PSQI score. CONCLUSIONS: Patients with HIV infection and not using evavirenz show an increased daytime sleepiness and a decreased quality of sleep. These findings could not be related to the immunological state of the patients. The only specific factor influencing daytime sleepiness in HIV infected patients is probably treatment with HAART. The most important factor determining sleepiness and sleep quality in HIV infected patients is depression which was found to be independent from the immunological state and HAART of the patients.

An improved PCR method for detection of HIV-1 proviral DNA of a wide range of subtypes and recombinant forms circulating globally.

Proviral DNAs are being measured increasingly as a marker of the efficacy of highly active anti-retroviral therapy (HAART) and is accepted for the early diagnosis of perinatal HIV-1 infections. This requires a standardized test which enables the detection of a wide range of subtypes worldwide including O, N and circulating recombinant forms (CRFs). Based on a previous publication, a PCR - Test for HIV-1 provirus detection in peripheral blood mononuclear cells (PBMCs) was developed. Blood samples from 80 individuals infected with HIV-1 and 20 persons negative for HIV-1&2 from Africa and Germany were tested for the presence of HIV-1 provirus DNA. The primer system used enables the detection of proviral DNA despite the high concentrations of human DNA. The limit of detection was determined to be 5 copies per 10(5) cells. All 20 samples from persons negative for HIV were negative for HIV-1 proviral DNA while provirus DNA was amplified from 76 of the 80 (95%) samples from persons infected with HIV. The amplified products were detected by gel-electrophoresis, flow cytometry and real-time PCR. All three detection systems provided the same results.

Severe leukoencephalopathy with fulminant cerebral edema reflecting immune reconstitution inflammatory syndrome during HIV infection: a case report.

INTRODUCTION: Immune reconstitution inflammatory syndrome is a well-known complication in HIV-infected patients after initiation of highly active antiretroviral therapy resulting in rapid CD4+ cell count recovery and suppression of viral load. Generally, immune reconstitution inflammatory syndrome is based on opportunistic infections, but rare cases of immune reconstitution inflammatory syndrome inducing demyelinization of the nervous system have also been observed. CASE PRESENTATION: A 37-year-old African woman with HIV infection diagnosed at 13 years of age was admitted to the emergency department after experiencing backache, severe headache, acute aphasia and psychomotor slowing for one week. Nine weeks earlier, highly active antiretroviral therapy in this patient had been changed because of loss of efficacy, and a rapid increase in CD4+ cell count and decrease of HIV viral load were observed. Magnetic resonance imaging of the brain showed extensive white matter lesions, and analysis of cerebrospinal fluid revealed an immunoreactive syndrome. Intensive investigations detected no opportunistic infections. A salvage therapy, including osmotherapy, corticosteroids and treatment of epileptic seizures, was performed, but the patient died from brainstem herniation 48 hours after admission. Neuropathologic examination of the brain revealed diffuse swelling, leptomeningeal infiltration by CD8 cells and enhancement of perivascular spaces by CD8+ cells. CONCLUSION: Immune reconstitution inflammatory syndrome in this form seems to represent a severe autoimmunologic disease of the brain with specific histopathologic findings. This form of immune reconstitution inflammatory syndrome did not respond to therapy, and extremely rapid deterioration led to death within two days. Immune reconstitution inflammatory syndrome may also occur as severe leukoencephalopathy with fulminant cerebral edema during HIV infection with rapid immune reconstitution.

Severe aseptic leucoencephalopathy as immune reconstitution inflammatory syndrome in Caucasian and African patients.

Immune reconstitution inflammatory syndrome (IRIS) after HAART may become manifest in form of aseptic severe leucoencephalopathy. All HIV-1-positive patients in this case series had widespread laboratory tests and follow-up MRI in order to investigate the course and the underlying pathophysiology of IRIS-associated leucoencephalopathy. All patients were treated with corticosteroids, in spite of additional immunosuppression. Three patients were successfully treated with corticosteroids and survived up to now, one died. A neuropathological examination was performed showing massive aseptic intraparenchymal and perivascular invasion of cytotoxic CD8 cells. It is assumed that IRIS-associated leucoencephalopathy is based on other preconditions in Africans and Caucasians.

A new affordable flow cytometry based method to measure HIV-1 viral load.

Most commercially available assays for diagnosis of HIV infection have shown shortcomings in the detection and quantification of rare genotypes of the virus. Most of the assays do not detect subtype O (outlier) and/or N (nonmajor, nonoutlier) or new circulating recombinant forms (CRFs), which are becoming more important in sub-Saharan Africa. Furthermore, the commonly available tests require costly measuring devices and expensive test kits, which are not easily affordable for developing countries. This study was designed to explore solutions to the problem of viral load assays in developing countries. Two forward primers, digoxygenin (DIG) and dinitrophenol (DNP) labeled, and one biotin (BIO) labeled reverse primer were used to amplify both, the HIV-1-5'LTR (long terminal repeat) region and an internal standard sequence. The two polymerase chain reaction (PCR)-products were captured by anti-DIG and anti-DNP antibody coated microparticles. Flow cytometric analyses were carried out after labeling with streptavidin-R-phycoerythrine. The primer system used recognized all HIV-1 subtypes. A coamplified internal standard warranted the functionality of the PCR and allows reproducible viral load measurements. Two drawbacks of current viral load measurements are overcome by the flow cytometry based test described hereof. First, all known worldwide relevant HIV-1 subtypes including subtypes O, N, and new CRFs are quantifiable with high sensitivity (50 to >1 x 10(6) copies per PCR). Second, the cost per test can be reduced to less than 12 US$ instead of the current 50-100 US$. Additionally, the test described in this report offers the possibility to perform complete monitoring program (CD4 T-cell count, CD4% and viral load) for the first time, with the same device for HIV-infected persons.

Simplified volumetric flow cytometry allows feasible and accurate determination of CD4 T lymphocytes in immunodeficient patients worldwide.

The determination of CD4 cells is of crucial clinical importance for patients with AIDS. However, the high costs involved represent limitations for CD4 cell counting in developing countries. In order to provide an affordable technique, we introduced a simplified volumetric counting (SVC) technique without sample manipulations and investigated it in a multicentre study. Blood samples from 434 healthy donors and immunodeficient patients were tested in eight hospital laboratories in Europe, Africa and Asia. CD4 cell counts were compared using in-house flow cytometric methods and the SVC technique. The SVC method was performed on a low-cost flow cytometer (CyFlow SL, Partec, Münster, Germany) after 15 min antibody incubation without pre-analytic manipulations, such as washing or erythrocyte lysing procedures. Linear regression analysis demonstrated a correlation of r=0.942 (Europe), r=0.952 (Africa) and r=0.989 (Asia) between the SVC technique and the in-house methods. Bland Altman plot analysis of all patient data showed a mean bias between the two methods of +26 CD4 cells in favour of the SVC technique (measured range: 6-1905 cells/microl; median CD4 cell count: 388/microl). Three centres used the FACS-count technique (Becton-Dickinson, San José, Calif., USA) as an in-house method dispensing with pre-analytic manipulations. The comparison of SVC and FACS-count method revealed a mean bias of +32 CD4 cells/microl (median CD4 cell count: 349/microl). The accuracy of the SVC was tested on standards with known CD4 cell counts (n=6) and was shown to be 95.2%. The low-cost device and the simplified no-lyse, no-wash test procedure reduces the costs per determination and facilitates the use of flow cytometry in developing countries.

Prevention of AIDS dementia by HAART does not depend on cerebrospinal fluid drug penetrance.

The impact of the cerebrospinal fluid (CSF) penetrance properties of different highly active antiretroviral therapy (HAART) regimes on cognitive processing in AIDS dementia is still undetermined. We therefore designed a retrospective cross-sectional and prospective longitudinal analysis of event-related potentials in HIV-infected patients with different combinations of HAART or without antiretroviral treatment. A total of 353 consecutive patients without secondary CNS manifestation of HIV infection were enrolled in the cross-sectional study and 135 consecutive patients without secondary CNS manifestations of HIV infection were enrolled in the longitudinal study. HAART in different combinations (n = 306) or no antiretroviral treatment (n = 47) was given for at least 6 months in the retrospective cross-sectional study. HAART in different combinations (n = 110) or no antiretroviral treatment (n = 25) was given for 1 year in the prospective longitudinal study. We evaluated the latency and amplitude of the P3 component of visually evoked event-related potentials and mean choice reaction time as measures of cognitive processing. Patients receiving HAART had decreased P3 latencies as compared to those patients not receiving HAART but P3 latency and P3 amplitude were not correlated with the amount of CSF penetrance of the different HAART combinations in either statistical analysis. However, mean choice reaction time was significantly correlated with the amount of CSF penetrance. In HIV-infected patients, the CSF penetrance properties of HAART do not have any significant influence on cognitive processing as measured by event-related potentials.

Ischaemic cerebrovascular events in HIV infection: a cohort study.

Several case reports and series described ischaemic cerebrovascular events in HIV infection. However, the exact prevalence and the clinical features of these events are unknown. We performed a cohort study on 772 consecutive HIV infected patients and evaluated the rate of transient ischaemic attacks (TIA) and of completed stroke. A total prevalence of 1.9% for TIA (0.8%) and stroke (1.2%) was calculated resulting in an annual incidence rate of 216 per 100000. The prevalence was highest in the later stages of the infection. Stroke patients had a poorer immunological state than the TIA and the cohort patients. Probable (n = 3) and possible (n = 2) vasculitis and cardiogenic embolism (n = 2) could be detected as aetiology, the remaining patients had a cryptogenic event. Our data suggest that ischaemic cerebrovascular events are more common in HIV infected patients than in the general population and that a part of these events might be caused by HIV associated vasculitis or vasculopathy.

Impact of highly active antiretroviral therapy on cognitive processing in HIV infection: cross-sectional and longitudinal studies of event-related potentials.

Patients with HIV infection often complain of cognitive disturbances, which can be related to AIDS dementia or HIV-associated encephalopathy (HIVE). We investigated the impact of highly active antiretroviral therapy (HAART) in comparison with other therapeutic regimens on the progression of these cognitive disturbances as measured by visual event-related potentials (ERP). In a cross-sectional study, 214 patients without secondary neuromanifestation of their infection were divided into four groups with respect to their treatment status for 1 year before examination: (1) without antiretroviral treatment, (2) zidovudine monotherapy, (3) zidovudine in combination with didanosine, zalcitabine, or lamivudine, and (4) HAART. In a prospective longitudinal study, we divided 54 patients into three groups: (1) without antiretroviral treatment, (2) zidovudine monotherapy, and (3) HAART. Latencies of the P2, N2, and P3 components and the amplitude of the P3 component were evaluated. A significant negative correlation between CD4(+) lymphocyte cell count and P3 latency was found in all patients (p < 0.004). In the cross-sectional study, P3 latency was significantly decreased in the HAART group as compared with patients with no antiretroviral treatment (p < 0.01). During the 1-year period of the prospective longitudinal study, the P3 latency significantly increased in patients with no antiretroviral treatment (p < 0.05) and significantly decreased in patients with HAART (p < 0.05). In summary, these results suggest that HAART has an improving therapeutic effect on cognitive processing in HIV-infected patients and is superior to zidovudine monotherapy or dual antiretroviral treatment. Because prolongation of ERP might in part reflect HIVE, we conclude that this condition represents an indication for HAART.