Development and validation of a clinical respiratory disease scoring system for guiding treatment decisions in veal calves using a Bayesian framework.

Active infectious bovine respiratory disease (BRD) is an infection of the airways that needs to be diagnosed correctly so that appropriate treatment can be initiated. The simplest and most practical test to detect active BRD in dairy calves raised for veal is the detection and interpretation of clinical signs by producers or technicians. However, the clinical scoring system currently available for veal calves lacks sensitivity and specificity, contributing to economic losses and high use of antimicrobials. An accurate and reliable batch-level test to detect active BRD is essential to tailor antimicrobial use and reduce economic losses in veal calves. The objective of this study was therefore to develop and validate a new veal calf respiratory clinical scoring system (VcCRS), including reliable clinical signs (cough, ear droop or head tilt) and increased rectal temperature to detect active BRD in batches of veal calves housed individually, and to describe the accuracy of the scoring system for identifying batches of veal calves to treat. During 2017 to 2018, clinical examination, thoracic ultrasonography (TUS) and a haptoglobin concentration (Hap) were prospectively performed on 800 veal calves housed individually in Québec, Canada. Deep nasopharyngeal swabs were performed on 250 veal calves. A Bayesian latent class model accounting for imperfect accuracy of TUS and Hap was used to obtain weights for the clinical signs and develop the VcCRS. The VcCRS was then validated externally in 3 separate data sets. Finally, the applicability of the VcCRS at batch level was determined. We found that calves with 2 of the following findings-cough, unilateral or bilateral ear droop or head tilt, or increased rectal temperature ≥39.7°C-were considered positive and had a 31% chance of having active BRD. Without at least 2 of these 2 findings, a calf had a 100% chance of not having active BRD. At the batch level, we found that a batch with ≥3 positive calves among 10 calves sampled 2 wk after arrival at the fattening unit had a 94% chance of having an active BRD prevalence ≥10%. A batch with <3 positive calves had a 95% chance of not having an active BRD prevalence ≥10%. In this study, we developed a simple individual and batch-level score that is reliable across examiners and performs effectively in the detection of active BRD in veal calves. The implementation of this VcCRS in the veal calf industry would promote the elaboration of a protocol tailoring antimicrobial use.

Association between transfer of passive immunity and health disorders in multisource commingled dairy calves raised for veal or other purposes: Systematic review and meta-analysis.

Inadequate transfer of passive immunity (ITPI) in newborn dairy calves remains an important risk factor for mortality and morbidity. Most available studies are focused on calves delivered and raised on the same farms. This setting is far different from calves transported and commingled from different farms to be raised as veal or for other purposes. The aim of this systematic review and meta-analysis was to describe the association between ITPI and important health outcomes (mortality, bovine respiratory disease, and diarrhea) in multisource commingled dairy calves raised for veal or other purposes. We searched studies through CAB abstracts (via CAB direct), PubMed, and Web of Science (via ISI) databases until September 2, 2021. Observational studies and randomized trials written in English or French assessing ITPI association with any of the selected outcomes were included. Young dairy calves transported to commercial facilities and explicitly stated as being raised for veal production or not (then considered as "other") were our populations of interest. If raw or adjusted data were available for ≥5 studies for a given outcome of interest, then random effect meta-analysis models were used to investigate ITPI effects on this outcome. Nineteen studies were selected from 6,221 abstracts retrieved in the initial search. We observed significantly higher odds of mortality in calves with ITPI compared with those with successful transfer of passive immunity [odds ratio (OR) = 2.46; 95% confidence interval (CI): 1.43-4.22, n = 8 studies]. Calves with ITPI had higher odds of diarrhea (OR = 3.03; 95% CI: 1.2-7.62, n = 7 studies). A significant publication bias toward publishing studies with positive results was found in studies reporting on bovine respiratory disease (n = 5 studies), which revealed nonsignificant associations after correction of publication bias (OR = 1.40; 95% CI: 0.77-2.6). Heterogeneity could not be thoroughly investigated for mortality and diarrhea due to the limited number of studies. Therefore, the pooled estimates of the random models should be interpreted with caution despite their robustness to sensitivity analyses. In this study, we also observed that multiple definitions for transfer of passive immunity and outcomes were used in the literature. Moreover, the raising system definition was often limited. There seems, therefore, to be a need for standardized definitions of these parameters, as well as a better description of systems used for multisource commingled dairy calves raised for veal production or other production purposes.

Aspergillus fumigatus and aspergillosis: From basics to clinics.

The airborne fungus Aspergillus fumigatus poses a serious health threat to humans by causing numerous invasive infections and a notable mortality in humans, especially in immunocompromised patients. Mould-active azoles are the frontline therapeutics employed to treat aspergillosis. The global emergence of azole-resistant A. fumigatus isolates in clinic and environment, however, notoriously limits the therapeutic options of mould-active antifungals and potentially can be attributed to a mortality rate reaching up to 100 %. Although specific mutations in CYP 51A are the main cause of azole resistance, there is a new wave of azole-resistant isolates with wild-type CYP 51A genotype challenging the efficacy of the current diagnostic tools. Therefore, applications of whole-genome sequencing are increasingly gaining popularity to overcome such challenges. Prominent echinocandin tolerance, as well as liver and kidney toxicity posed by amphotericin B, necessitate a continuous quest for novel antifungal drugs to combat emerging azole-resistant A. fumigatus isolates. Animal models and the tools used for genetic engineering require further refinement to facilitate a better understanding about the resistance mechanisms, virulence, and immune reactions orchestrated against A. fumigatus. This review paper comprehensively discusses the current clinical challenges caused by A. fumigatus and provides insights on how to address them.

Evaluation of inter-rater agreement of the clinical signs used to diagnose bovine respiratory disease in individually housed veal calves.

In dairy calves raised for veal, typical clinical signs of bovine respiratory disease (BRD) are ocular discharge, nasal discharge, ear droop or head tilt, abnormal respiration, cough, and increased rectal temperature. Despite the existence of several clinical scoring systems, there are few studies on the variability of human recognition of individual BRD clinical signs. The objective of this study was therefore to assess the inter-rater agreement of BRD clinical signs in veal calves. We hypothesized that BRD clinical signs were not detected equally between veterinarians, technicians, and producers of the veal industry and that some clinical signs have higher inter-rater agreement than others. During 2017-2018, we prospectively recorded 524 videos of physical examinations of random veal calves from 48 different batches in Québec, Canada. A researcher, not involved in the inter-rater assessment, classified each video as presence/absence of each BRD clinical sign except rectal temperature. For each of the 5 clinical signs, 15 videos with and 15 videos without the clinical signs were randomly selected to avoid kappa paradoxes. Those 30 videos were then presented in a random order to experienced raters of BRD in veal calves: 6 veterinarians, 6 technicians, and 6 producers. The raters assessed the clinical signs using scores based on the Wisconsin and California scoring system with modifications (0 = absent, 1 = mild, 2 = moderate, 3 = severe for nasal discharge, ocular discharge, and ear droop or head tilt; and 0 = absent, 1 = moderate, 2 = severe for abnormal respiration and induced cough). We used median percentage agreement (Pa), median Cohen's kappa (κ), and Gwet's agreement coefficient 1 (AC1) to assess inter-rater agreement. The effect of scale combination was also tested to determine the optimal combination (4-scale 0/1/2/3 vs. 3-scale 0/1/2 vs. 2-scale 0/1,2,3; 0,1/2,3; or 0/1,2). The differences of inter-rater agreement between veterinarians, technicians, and producers were estimated by a Wilcoxon rank-sum test. The 2-scale combination (0,1/2,3 or 0/1,2) had the highest inter-rater agreement for all clinical signs. With this combination, induced cough was the clinical sign with the highest inter-rater agreement (Pa = 0.93; κ = 0.79; AC1 = 0.87) and abnormal respiration was the sign with the lowest inter-rater agreement (Pa = 0.77; κ = 0.20; AC1 = 0.74). According to Pa and AC1 values, the 2-scale inter-rater agreement of the 5 clinical signs was good (value > 0.6). According to κ, only ear droop or head tilt and induced cough had a substantial 2-scale inter-rater agreement (κ > 0.6). In general, the 2-scale inter-rater agreement was better among veterinarians than among technicians and producers, except for the ear droop/head tilt, where agreement was better among producers. We concluded that with severity scores assessed on a scale of 2 (0,1/2,3 or 0/1,2), the inter-rater agreement of BRD clinical signs was variable according to the sign in veal calves. BRD clinical signs were not detected equally between veterinarians, technicians, and producers of the veal industry. Future research could determine if this discrepancy could be improved by standardization training.

Hot under the collar: A 14-year association between temperature and violent behavior across 436 U.S. counties.

OBJECTIVES: Violent behavior is influenced by individual and societal characteristics, but the role of environmental factors is less understood. Our aims were to use national-level data to identify the association between criminal behavior and short-term temperature conditions, including the departure of daily temperatures from normal conditions. METHODS: We conducted a multi-stage hierarchical time-series model across 436 U.S. counties and 14-years representing 100.4 million people to investigate the association between daily mean temperature and daily mean temperatures departing from normal conditions with violent and non-violent crime counts. First-stage comparisons were made within counties to control for population and geographic heterogeneities, while a second stage combined estimates. We evaluated differences in risk based on county sociodemographic characteristics and estimated non-linear exposure-response relationships. RESULTS: We observed a total of 9.0 million violent crimes and 20.9 million non-violent property crimes between 2000 through 2013. We estimated that each 10 °C increase in daily temperature or daily departure from long-term normal temperatures were associated with 11.92% (95% PI: 11.57, 12.27) and 10.37% (95% PI: 10.05, 10.69) increase in the risk of violent crime, respectively. Similar, but lower in magnitude trends, were observed for property crime risks. We found that crime risk plateaus and decreases at high daily temperatures, but for temperatures departing from normal, the association with crime increased linearly. Seasonal variations showed that anomalously warm temperatures days during cool months had the greatest risk. CONCLUSIONS: Our study revealed an association between higher temperatures and high departure from normal temperatures with both violent and non-violent crime risk, regardless of community-type. However, our findings on seasonal and daily trends suggest that daily mean temperature may impact crime by affecting routine activities and behavior, as opposed to a temperature-aggression relationship. These results may advance public response and planning to prevent violent behavior.

Feasibility of Patient-Reported Outcomes Measurement Information System (PROMIS®) computerized adaptive tests in systemic lupus erythematosus outpatients.

Objective The aims of this study were to assess the feasibility of administering Patient-Reported Outcomes Measurement Information System (PROMIS®) computerized adaptive tests (CATs) to outpatients with systemic lupus erythematosus (SLE). Methods Adults with SLE were recruited during routine outpatient visits at an SLE Center of Excellence. Participants completed 14 PROMIS CATs and provided feedback on their experience. Differences in socio-demographic and clinical characteristics between participants and non-participants were evaluated. Results A total of 204 (86%) of 238 socioeconomically and racially diverse SLE patients completed PROMIS CATs. There were no significant differences between participants and non-participants. Time constraints were cited most frequently as reasons for non-participation. More than 75% of individuals submitted positive comments, including approval of the content and format of questions, and the survey's promotion of self-reflection. A minority of participants cited challenges, most often related to question phrasing (8%) and technical difficulties (6%). Conclusions The administration of PROMIS CATs was feasible and positively received in a diverse cohort of SLE outpatients. Neither socio-demographic nor disease characteristics were significant barriers to successful completion of PROMIS CATs. PROMIS CATs have great potential for efficiently measuring important patient-centered outcomes in routine clinical care of a wide range of SLE patients.

A randomised clinical trial of a metaphylactic treatment with tildipirosin for bovine respiratory disease in veal calves.

BACKGROUND: Bovine respiratory disease (BRD) is a major problem in veal calf rearing units. The objective of this randomised clinical trial was to assess the effectiveness of tildipirosin as a metaphylactic treatment in veal calves on the number of BRD treatments, lung consolidation on thoracic ultrasonography (TUS) and average daily gain (ADG). A total of 209 veal calves from a pre-weaning fattening unit were randomly allocated to receive one of two treatments (tildipirosin 4 mg/kg, subcutaneously, n = 109; placebo 0.9% saline, subcutaneously, n = 100) at day 12 after entry in the pre-weaned unit. The calves were followed for a 70-day period. Occurrence of mortality and BRD treatments were recorded during the pre-weaning period. At days 1, 12 and 30, TUS and clinical scores were performed and ADG was measured during the first and second months of feeding. RESULTS: The use of a metaphylactic treatment of tildipirosin 12 days after arrival of the veal calves was not associated with the number of BRD treatments performed by the producer, ultrasonographic lung consolidation or weight gain (P < 0,05). In this cohort of calves, the proportion of calves treated for BRD by the producer was low at 14% (29/209). However, 13% (26/209) of calves included in the study already had ultrasonographic lung consolidation lesions 12 days after their arrival, which was before treatment time, and 27% (56/209) had lung consolidation at day 30. CONCLUSION: In this study population with a low BRD prevalence, we were not able to detect any benefit of tildipirosin as a metaphylactic treatment of BRD at day 12 after arrival based on BRD treatments, TUS, and ADG.

Myelin basic protein induces inflammatory mediators from primary human endothelial cells and blood-brain barrier disruption: implications for the pathogenesis of multiple sclerosis.

AIM: Multiple sclerosis (MS) is an autoimmune disease of the central nervous system, characterized by demyelination of white matter, loss of myelin forming oligodendrocytes, changes in the blood-brain barrier (BBB) and leucocyte infiltration. Myelin basic protein (MBP) is a component of the myelin sheath. Degradation of myelin is believed to be an important step that leads to MS pathology. Transmigration of leucocytes across the vasculature, and a compromised BBB participate in the neuroinflammation of MS. We examined the expression and regulation of the chemokine (C-C motif) ligand 2 (CCL2) and the cytokine interleukin-6 (IL-6) in human endothelial cells (EC), a component of the BBB, after treatment with MBP. METHODS: EC were treated with full-length MBP. CCL2 and IL-6 protein were determined by ELISA. Western blot analysis was used to determine signalling pathways. A BBB model was treated with MBP and permeability was assayed using albumin conjugated to Evan's blue dye. The levels of the tight junction proteins occludin and claudin-1, and matrix metalloprotease (MMP)-2 were assayed by Western blot. RESULTS: MBP significantly induced CCL2 and IL-6 protein from EC. This induction was partially mediated by the p38 MAPK pathway as there was phosphorylation after MBP treatment. MBP treatment of a BBB model caused an increase in permeability that correlated with a decrease in occludin and claudin-1, and an induction of MMP2. CONCLUSION: These data demonstrate that MBP induces chemotactic and inflammatory mediators. MBP also alters BBB permeability and tight junction expression, indicating additional factors that may contribute to the BBB breakdown characteristic of MS.

Fine-mapping in African-American women confirms the importance of the 10p12 locus to sarcoidosis.

Sarcoidosis is a chronic granulomatous disease with a wide spectrum of symptoms. Genome-wide association studies in European populations have reported significant associations between sarcoidosis and single-nucleotide polymorphisms (SNPs) located in the intergenic region between the C10ORF67 and OTUD1 genes on chromosome 10p12, and the ANXA11 gene (chromosome 10q22). We carried out fine-mapping at 10p12 and 10q22 to assess associations of genetic variants in these regions with sarcoidosis risk in African-American women, based on 486 sarcoidosis cases and 943 age- and geography-matched controls in a nested case-control study within the Black Women's Health Study. There were no significant associations with variants of the ANXA11 gene (P=0.17). Haplotypic analyses of the C10ORF67-OTUD1 intergenic region revealed a strong inverse association of the variants rs1398024 and rs11013452 with sarcoidosis (odds ratio=0.52; P=0.01). Both SNPs are located inside an ∼300 kb low recombination region of chromosome 10p12, suggesting that both SNPs are tagging the same causal variant. Our top SNP (rs11013452) is located inside a smaller linkage disequilibrium block in HapMap YRI, further narrowing the position of the causal SNP to a region of ~8 kb on chromosome 10p12. The present findings confirm the potential importance of the 10p12 locus in the etiology of sarcoidosis.

Rad52 function prevents chromosome loss and truncation in Candida albicans.

RAD52 is required for almost all recombination events in Saccharomyces cerevisiae. We took advantage of the heterozygosity of HIS4 in the Candida albicans SC5314 lineage to study the role of Rad52 in the genomic stability of this important fungal pathogen. The rate of loss of heterozygosity (LOH) at HIS4 in rad52-ΔΔ strains was ∼10(-3) , at least 100-fold higher than in Rad52(+) strains. LOH of whole chromosome 4 or truncation of the homologue that carries the functional HIS4 allele was detected in all 80 rad52-ΔΔ His auxotrophs (GLH -GL lab His(-)) obtained from six independent experiments. Isolates that had undergone whole chromosome LOH, presumably due to loss of chromosome, carried two copies of the remaining homologue. Isolates with truncations carried centric fragments of broken chromosomes healed by de novo telomere addition. GLH strains exhibited variable degrees of LOH across the genome, including two strains that became homozygous for all the heterozygous markers tested. In addition, GLH strains exhibited increased chromosomal instability (CIN), which was abolished by reintroduction of RAD52. CIN of GLH isolates is reminiscent of genomic alterations leading to cancer in human cells, and support the mutator hypothesis in which a mutator mutation or CIN phenotype facilitate more mutations/aneuploidies.

Metabolism of formycin B by Leishmania amastigotes in vitro. Comparative metabolism in infected and uninfected human macrophages.

Formycin B is metabolized by cutaneous Leishmania amastigotes within cultured human macrophages to give formycin B 5'-monophosphate and formycin A 5'-mono-, di-, and triphosphates. Formycin A is also incorporated into RNA. The activity of formycin B against amastigotes was correlated with the levels of formycin A metabolites formed in the parasites. Uninfected macrophages also convert formycin B into the same products, but the levels are markedly lower than those seen in infected macrophages. The results suggest that a sufficient therapeutic index exists to warrant consideration of formycin B as an anti-leishmanial drug in humans.

Structure and expression of human germline VH transcripts.

The human VH5 family consists of two functional genes and one pseudogene. We have found a novel 1.2-kb VH5 gene transcript in normal fetal liver and cord blood and in transformed B lineage cells. VH5-positive cDNA clones were isolated from precursor B acute lymphoblastic leukemia, B chronic lymphoblastic leukemia, Epstein-Barr Virus-transformed B cell lines, and cord blood, and were identified as transcripts of unrearranged VH5 genes (germline transcripts). The cDNA clones were derived from both functional and pseudo-VH5 genes. Most germline transcripts appear to initiate at the normal VH promoter and are cleaved and polyadenylated at sites several hundred bases downstream of the VH5 coding region. Correct splicing of the leader intron was observed in all clones. In functional and pseudo-VH5 cDNAs, an open translational reading frame extends from the leader to a termination codon in the nonamer. Only limited polymorphisms were observed in the coding as well as flanking regions of the VH5 transcripts. Functional and pseudo-VH5 transcripts and previously identified murine germline VHJ558 transcripts are discussed.

Early human IgH gene assembly in Epstein-Barr virus-transformed fetal B cell lines. Preferential utilization of the most JH-proximal D segment (DQ52) and two unusual VH-related rearrangements.

We have analyzed the phenotypic characteristics and IgH gene rearrangements in a panel of EBV-transformed B lineage cell lines from human fetal liver and bone marrow. Some lines contained only populations of immature, Ig- Be cells, while others contained mixed populations of mature and immature B cells. The majority of identifiable IgH rearrangements involved joining of the most JH-proximal D segment, DQ52, to various JH segments, implying that DQ52 is a preferred target for initial DJH rearrangements. Three other rearrangements involving VH-related sequences were also characterized. Two involved VHDJH joining using VH3 genes, although one of these had a very unusual DJH structure. The third consisted of inverted 3' signal sequences and flanking regions of a VH4 gene appended to a JH. The mechanisms by which the later rearrangement could have occurred and its potential physiological significance are discussed.

Tumor necrosis factor alpha induces adhesion molecule expression on human fetal astrocytes.

Leukocyte adhesion molecules on endothelial cells of the blood-brain barrier may participate in the entry of leukocytes into the central nervous system. Because astrocytes are also a component of the blood-brain barrier and have been associated with inflammation, we studied the ability of astrocytes to express leukocyte adhesion molecules using Northern blot and immunocytochemical techniques. Astrocytes treated with the proinflammatory cytokine tumor necrosis factor alpha (TNF) expressed messenger RNA for the adhesion molecules E-selectin, vascular cell adhesion molecule 1, and intercellular adhesion molecule 1, as well as their corresponding proteins. In addition, TNF-treated astrocytes expressed a monocyte adhesion protein identified by our laboratory, recognized by the monoclonal antibody IG9. These results indicate that under inflammatory conditions in the central nervous system, such as multiple sclerosis and acquired immune deficiency syndrome, astrocyte expression of adhesion molecules may facilitate the migration of leukocytes and contribute to the disease process.

Comparison between thoracic ultrasonography and thoracic radiography for the detection of thoracic lesions in dairy calves using a two-stage Bayesian method.

Infectious bronchopneumonia is a lower respiratory tract disease with major economic consequences in dairy calves. Thoracic radiography (TR) and thoracic ultrasonography (TUS) are two imaging diagnostic procedures available in bovine medicine for identifying thoracic lesions. However, no study has investigated whether one of these tests is superior to the other or if they provide comparable results for the detection of thoracic lesions in calves. The objective of this study was therefore to estimate and to compare the performances of TUS and TR for the detection of thoracic lesions in dairy calves. A prospective cross-sectional study was performed in a hospital setting. A total of 50 calves (≥7 days old; ≤100 kg; standing; pCO2 ≥ 53 mmHg; any reason of presentation) were enrolled. Every calf underwent TUS and TR. Only calves with thoracic lesions on TUS and/or TR were controlled by thoracic computed tomography (CT) (the gold standard). Calves without lesions were not controlled by CT. A two-stage Bayesian framework was used. The sensitivities (Se) and specificities (Sp) of both tests individually and used in series or parallel were estimated. The Se and Sp of TUS were 0.81 (95 % BCI (Bayesian Credible Interval): 0.65; 0.92) and 0.90 (95 % BCI: 0.81; 0.96), respectively. The Se and Sp of TR were 0.86 (95 % BCI: 0.62; 0.99) and 0.89 (95 % BCI: 0.67; 0.99), respectively. This study did not reveal any differences between both tests. Using TUS and TR in series was more specific than using both tests in parallel. The performances of TUS alone were not different from the performances of both tests in series or in parallel. In conclusion, TUS and TR were equivalent in detecting thoracic lesions in this study. Using TUS alone allowed an accurate detection of thoracic lesions in dairy calves. Further studies enrolling a larger sample (> 400 calves) and allowing adequate power to be achieved would be necessary to confirm these results.

HIV Neuropathogenesis in the Presence of a Disrupted Dopamine System.

Antiretroviral therapy (ART) has transformed HIV into a chronic condition, lengthening and improving the lives of individuals living with this virus. Despite successful suppression of HIV replication, people living with HIV (PLWH) are susceptible to a growing number of comorbidities, including neuroHIV that results from infection of the central nervous system (CNS). Alterations in the dopaminergic system have long been associated with HIV infection of the CNS. Studies indicate that changes in dopamine concentrations not only alter neurotransmission, but also significantly impact the function of immune cells, contributing to neuroinflammation and neuronal dysfunction. Monocytes/macrophages, which are a major target for HIV in the CNS, are responsive to dopamine. Therefore, defining more precisely the mechanisms by which dopamine acts on these cells, and the changes in cellular function elicited by this neurotransmitter are necessary to develop therapeutic strategies to treat neuroHIV. This is especially important for vulnerable populations of PLWH with chemically altered dopamine concentrations, such as individuals with substance use disorder (SUD), or aging individuals using dopamine-altering medications. The specific neuropathologic and neurocognitive consequences of increased CNS dopamine remain unclear. This is due to the complex nature of HIV neuropathogenesis, and logistical and technical challenges that contribute to inconsistencies among cohort studies, animal models and in vitro studies, as well as lack of demographic data and access to human CNS samples and cells. This review summarizes current understanding of the impact of dopamine on HIV neuropathogenesis, and proposes new experimental approaches to examine the role of dopamine in CNS HIV infection. Graphical abstract HIV Neuropathogenesis in the Presence of a Disrupted Dopamine System. Both substance abuse disorders and the use of dopaminergic medications for age-related diseases are associated with changes in CNS dopamine concentrations and dopaminergic neurotransmission. These changes can lead to aberrant immune function, particularly in myeloid cells, which contributes to the neuroinflammation, neuropathology and dysfunctional neurotransmission observed in dopamine-rich regions in HIV+ individuals. These changes, which are seen despite the use antiretroviral therapy (ART), in turn lead to further dysregulation of the dopamine system. Thus, in individuals with elevated dopamine, the bi-directional interaction between aberrant dopaminergic neurotransmission and HIV infection creates a feedback loop contributing to HIV associated neurocognitive dysfunction and neuroHIV. However, the distinct contributions and interactions made by HIV infection, inflammatory mediators, ART, drugs of abuse, and age-related therapeutics are poorly understood. Defining more precisely the mechanisms by which these factors influence the development of neurological disease is critical to addressing the continued presence of neuroHIV in vulnerable populations, such as HIV-infected older adults or drug abusers. Due to the complexity of this system, understanding these effects will require a combination of novel experimental modalities in the context of ART. These will include more rigorous epidemiological studies, relevant animal models, and in vitro cellular and molecular mechanistic analysis.

Tunneling nanotubes (TNT) are induced by HIV-infection of macrophages: a potential mechanism for intercellular HIV trafficking.

Cell to cell communication is essential for the organization/coordination of multicellular systems and cellular development. Cellular communication is mediated by soluble factors, including growth factors, neurotransmitters, cytokines/chemokines, gap junctions, and the recently described tunneling nanotubes (TNT). TNT are long cytoplasmatic bridges that enable long range directed communication between cells. The proposed function for TNT is the cell-to-cell transfer of large cellular structures such as vesicles and organelles. We demonstrate that HIV-infection of human macrophages results in an increased number of TNT, and show HIV particles within these structures. We propose that HIV "highjacks" TNT communication to spread HIV through an intercellular route between communicated cells, contributing to the pathogenesis of AIDS.