Plasma phosphorylated tau 217 and phosphorylated tau 181 as biomarkers in Alzheimer's disease and frontotemporal lobar degeneration: a retrospective diagnostic performance study

Background: Plasma tau phosphorylated at threonine 217 (p-tau217) and plasma tau phosphorylated at threonine 181 (p-tau181) are associated with Alzheimer's disease tau pathology. We compared the diagnostic value of both biomarkers in cognitively unimpaired participants and patients with a clinical diagnosis of mild cognitive impairment, Alzheimer's disease syndromes, or frontotemporal lobar degeneration (FTLD) syndromes. / Methods: In this retrospective multicohort diagnostic performance study, we analysed plasma samples, obtained from patients aged 18–99 years old who had been diagnosed with Alzheimer's disease syndromes (Alzheimer's disease dementia, logopenic variant primary progressive aphasia, or posterior cortical atrophy), FTLD syndromes (corticobasal syndrome, progressive supranuclear palsy, behavioural variant frontotemporal dementia, non-fluent variant primary progressive aphasia, or semantic variant primary progressive aphasia), or mild cognitive impairment; the participants were from the University of California San Francisco (UCSF) Memory and Aging Center, San Francisco, CA, USA, and the Advancing Research and Treatment for Frontotemporal Lobar Degeneration Consortium (ARTFL; 17 sites in the USA and two in Canada). Participants from both cohorts were carefully characterised, including assessments of CSF p-tau181, amyloid-PET or tau-PET (or both), and clinical and cognitive evaluations. Plasma p-tau181 and p-tau217 were measured using electrochemiluminescence-based assays, which differed only in the biotinylated antibody epitope specificity. Receiver operating characteristic analyses were used to determine diagnostic accuracy of both plasma markers using clinical diagnosis, neuropathological findings, and amyloid-PET and tau-PET measures as gold standards. Difference between two area under the curve (AUC) analyses were tested with the Delong test. / Findings: Data were collected from 593 participants (443 from UCSF and 150 from ARTFL, mean age 64 years [SD 13], 294 [50%] women) between July 1 and Nov 30, 2020. Plasma p-tau217 and p-tau181 were correlated (r=0·90, p<0·0001). Both p-tau217 and p-tau181 concentrations were increased in people with Alzheimer's disease syndromes (n=75, mean age 65 years [SD 10]) relative to cognitively unimpaired controls (n=118, mean age 61 years [SD 18]; AUC=0·98 [95% CI 0·95–1·00] for p-tau217, AUC=0·97 [0·94–0·99] for p-tau181; pdiff=0·31) and in pathology-confirmed Alzheimer's disease (n=15, mean age 73 years [SD 12]) versus pathologically confirmed FTLD (n=68, mean age 67 years [SD 8]; AUC=0·96 [0·92–1·00] for p-tau217, AUC=0·91 [0·82–1·00] for p-tau181; pdiff=0·22). P-tau217 outperformed p-tau181 in differentiating patients with Alzheimer's disease syndromes (n=75) from those with FTLD syndromes (n=274, mean age 67 years [SD 9]; AUC=0·93 [0·91–0·96] for p-tau217, AUC=0·91 [0·88–0·94] for p-tau181; pdiff=0·01). P-tau217 was a stronger indicator of amyloid-PET positivity (n=146, AUC=0·91 [0·88–0·94]) than was p-tau181 (n=214, AUC=0·89 [0·86–0·93]; pdiff=0·049). Tau-PET binding in the temporal cortex was more strongly associated with p-tau217 than p-tau181 (r=0·80 vs r=0·72; pdiff<0·0001, n=230). / Interpretation: Both p-tau217 and p-tau181 had excellent diagnostic performance for differentiating patients with Alzheimer's disease syndromes from other neurodegenerative disorders. There was some evidence in favour of p-tau217 compared with p-tau181 for differential diagnosis of Alzheimer's disease syndromes versus FTLD syndromes, as an indication of amyloid-PET-positivity, and for stronger correlations with tau-PET signal. Pending replication in independent, diverse, and older cohorts, plasma p-tau217 and p-tau181 could be useful screening tools to identify individuals with underlying amyloid and Alzheimer's disease tau pathology. / Funding: US National Institutes of Health, State of California Department of Health Services, Rainwater Charitable Foundation, Michael J Fox foundation, Association for Frontotemporal Degeneration, Alzheimer's Association

Biomarkers for Clinical and Incipient Tuberculosis: Performance in a TB-Endemic Country

Simple biomarkers are required to identify TB in both HIV(-)TB(+) and HIV(+)TB(+) patients. Earlier studies have identified the M. tuberculosis Malate Synthase (MS) and MPT51 as immunodominant antigens in TB patients. One goal of these investigations was to evaluate the sensitivity and specificity of anti-MS and -MPT51 antibodies as biomarkers for TB in HIV(-)TB(+) and HIV(+)TB(+) patients from a TB-endemic setting. Earlier studies also demonstrated the presence of these biomarkers during incipient subclinical TB. If these biomarkers correlate with incipient TB, their prevalence should be higher in asymptomatic HIV(+) subjects who are at a high-risk for TB. The second goal was to compare the prevalence of these biomarkers in asymptomatic, CD4(+) T cell-matched HIV(+)TB(-) subjects from India who are at high-risk for TB with similar subjects from US who are at low-risk for TB.Anti-MS and -MPT51 antibodies were assessed in sera from 480 subjects including PPD(+) or PPD(-) healthy subjects, healthy community members, and HIV(-)TB(+) and HIV(+)TB(+) patients from India. Results demonstrate high sensitivity (approximately 80%) of detection of smear-positive HIV(-)TB(+) and HIV(+)TB(+) patients, and high specificity (>97%) with PPD(+) subjects and endemic controls. While approximately 45% of the asymptomatic HIV(+)TB(-) patients at high-risk for TB tested biomarker-positive, >97% of the HIV(+)TB(-) subjects at low risk for TB tested negative. Although the current studies are hampered by lack of knowledge of the outcome, these results provide strong support for the potential of these biomarkers to detect incipient, subclinical TB in HIV(+) subjects.These biomarkers provide high sensitivity and specificity for TB diagnosis in a TB endemic setting. Their performance is not compromised by concurrent HIV infection, site of TB and absence of pulmonary manifestations in HIV(+)TB(+) patients. Results also demonstrate the potential of these biomarkers for identifying incipient subclinical TB in HIV(+)TB(-) subjects at high-risk for TB

Transcriptional Reprogramming in Nonhuman Primate (Rhesus Macaque) Tuberculosis Granulomas

In response to Mtb infection, the host remodels the infection foci into a dense mass of cells known as the granuloma. The key objective of the granuloma is to contain the spread of Mtb into uninfected regions of the lung. However, it appears that Mtb has evolved mechanisms to resist killing in the granuloma. Profiling granuloma transcriptome will identify key immune signaling pathways active during TB infection. Such studies are not possible in human granulomas, due to various confounding factors. Nonhuman Primates (NHPs) infected with Mtb accurately reflect human TB in clinical and pathological contexts.We studied transcriptomics of granuloma lesions in the lungs of NHPs exhibiting active TB, during early and late stages of infection. Early TB lesions were characterized by a highly pro-inflammatory environment, expressing high levels of immune signaling pathways involving IFNgamma, TNFalpha, JAK, STAT and C-C/C-X-C chemokines. Late TB lesions, while morphologically similar to the early ones, exhibited an overwhelming silencing of the inflammatory response. Reprogramming of the granuloma transcriptome was highly significant. The expression of approximately two-thirds of all genes induced in early lesions was later repressed.The transcriptional characteristics of TB granulomas undergo drastic changes during the course of infection. The overwhelming reprogramming of the initial pro-inflammatory surge in late lesions may be a host strategy to limit immunopathology. We propose that these host profiles can predict changes in bacterial replication and physiology, perhaps serving as markers for latency and reactivation

N-Terminal Gly224–Gly411 Domain in Listeria Adhesion Protein Interacts with Host Receptor Hsp60

Listeria adhesion protein (LAP) is a housekeeping bifunctional enzyme consisting of N-terminal acetaldehyde dehydrogenase (ALDH) and C-terminal alcohol dehydrogenase (ADH). It aids Listeria monocytogenes in crossing the epithelial barrier through a paracellular route by interacting with its host receptor, heat shock protein 60 (Hsp60). To gain insight into the binding interaction between LAP and Hsp60, LAP subdomain(s) participating in the Hsp60 interaction were investigated.Using a ModBase structural model, LAP was divided into 4 putative subdomains: the ALDH region contains N1 (Met(1)-Pro(223)) and N2 (Gly(224)-Gly(411)), and the ADH region contains C1 (Gly(412)-Val(648)) and C2 (Pro(649)-Val(866)). Each subdomain was cloned and overexpressed in Escherichia coli and purified. Purified subdomains were used in ligand overlay, immunofluorescence, and bead-based epithelial cell adhesion assays to analyze each domain's affinity toward Hsp60 protein or human ileocecal epithelial HCT-8 cells.The N2 subdomain exhibited the greatest affinity for Hsp60 with a K(D) of 9.50±2.6 nM. The K(D) of full-length LAP (7.2±0.5 nM) to Hsp60 was comparable to the N2 value. Microspheres (1 µm diameter) coated with N2 subdomain showed significantly (P<0.05) higher binding to HCT-8 cells than beads coated with other subdomains and this binding was inhibited when HCT-8 cells were pretreated with anti-Hsp60 antibody to specifically block epithelial Hsp60. Furthermore, HCT-8 cells pretreated with purified N2 subdomain also reduced L. monocytogenes adhesion by about 4 log confirming its involvement in interaction with epithelial cells.These data indicate that the N2 subdomain in the LAP ALDH domain is critical in initiating interaction with mammalian cell receptor Hsp60 providing insight into the molecular mechanism of pathogenesis for the development of potential anti-listerial control strategies

New approaches in the diagnosis and treatment of latent tuberculosis infection

With nearly 9 million new active disease cases and 2 million deaths occurring worldwide every year, tuberculosis continues to remain a major public health problem. Exposure to Mycobacterium tuberculosis leads to active disease in only ~10% people. An effective immune response in remaining individuals stops M. tuberculosis multiplication. However, the pathogen is completely eradicated in ~10% people while others only succeed in containment of infection as some bacilli escape killing and remain in non-replicating (dormant) state (latent tuberculosis infection) in old lesions. The dormant bacilli can resuscitate and cause active disease if a disruption of immune response occurs. Nearly one-third of world population is latently infected with M. tuberculosis and 5%-10% of infected individuals will develop active disease during their life time. However, the risk of developing active disease is greatly increased (5%-15% every year and ~50% over lifetime) by human immunodeficiency virus-coinfection. While active transmission is a significant contributor of active disease cases in high tuberculosis burden countries, most active disease cases in low tuberculosis incidence countries arise from this pool of latently infected individuals. A positive tuberculin skin test or a more recent and specific interferon-gamma release assay in a person without overt signs of active disease indicates latent tuberculosis infection. Two commercial interferon-gamma release assays, QFT-G-IT and T-SPOT.TB have been developed. The standard treatment for latent tuberculosis infection is daily therapy with isoniazid for nine months. Other options include therapy with rifampicin for 4 months or isoniazid + rifampicin for 3 months or rifampicin + pyrazinamide for 2 months or isoniazid + rifapentine for 3 months. Identification of latently infected individuals and their treatment has lowered tuberculosis incidence in rich, advanced countries. Similar approaches also hold great promise for other countries with low-intermediate rates of tuberculosis incidence

Does the fluence map editing in electronic tissue compensator improve dose homogeneity in bilateral field plan of head and neck patients?

The purpose of this study was to evaluate the effect of fluence map editing in electronic tissue compensator (ETC) on the dose homogeneity for head and neck cancer patients. Treatment planning using 6-MV X-rays and bilateral field arrangement employing ETC was carried out on the computed tomography (CT) datasets of 20 patients with head and neck cancer. All the patients were planned in Varian Eclipse three-dimensional treatment planning system (3DTPS) with dynamic multileaf collimator (DMLC). The treatment plans, with and without fluence editing, was compared and the effect of pre-editing and post-editing the fluence maps in the treatment field was evaluated. The skin dose was measured with thermoluminescent dosimeters (TLDs) and was compared with the skin dose estimated by TPS. The mean percentage volume of the tissue receiving at least 107% of the prescription dose was 5.4 (range 1.5-10; SD 2.4). Post-editing fluence map showed that the mean percentage volume of the tissue receiving at least 107% of the prescription dose was 0.47 (range 0.1-0.9; SD 0.3). The mean skin dose measured with TLD was found to be 74% (range 71-80%) of the prescribed dose while the TPS showed the mean skin dose as 85% (range 80-90%). The TPS overestimated the skin dose by 11%. Fluence map editing thus proved to be a potential tool for improving dose homogeneity in head and neck cancer patients planned with ETC, thus reducing the hot spots in the treatment region as well. The treatment with ETC is feasible with DMLC and does not take any additional time for setup or delivery. The method used to edit the fluence maps is simple and time efficient. Manual control over a plan is essential to create the best treatment plan possible

Multileaf collimator transmission from the first Hi-Art II helical tomotherapy machine in India

The purpose of this study was to measure the multileaf collimator (MLC) transmission from the first Hi-Art II tomotherapy machine installed at the Advanced Center for Treatment, Research, and Education in Cancer (ACTREC). The MLC transmission was measured with an A1SL ion chamber and the radiographic extended dose range (EDR2) film in virtual water slabs at 1.5-cm depth with a source-to-surface distance of 85 cm. The MLC transmission was measured for 30 s with all leaves open and for 360 s with all leaves closed. The movable jaws were set to the calibration field size of 5 x 40 cm at isocenter. The MLC transmission was found to be 0.3% with the ion chamber and 0.32% with the film. Thus, the MLC transmission value was found well within the manufacturer tolerance of 0.5%. MLC can safely be used for the beam modulation during intensity-modulated radiotherapy (IMRT) to deliver accurate doses to the patients

Characterization of metal oxide field-effect transistors for first helical tomotherapy Hi-Art II unit in India

Purpose : To characterize metal oxide semiconductor field-effect transistors (MOSFETs) for a 6-MV photon beam with a first helical tomotherapy Hi-Art II unit in India. Materials and Methods : Standard sensitivity MOSFETs were first calibrated and then characterized for reproducibility, field size dependence, angular dependence, fade effects, and temperature dependence. The detector sensitivity was estimated for static as well as rotational modes for three jaw settings (1.0 cm 7 40 cm, 2.5 cm 7 40 cm, and 5 cm 7 40 cm) at 1.5-cm depth with a source-to-axis distance (SAD) of 85 cm in virtual water slabs. The A1SL ion chamber and thermoluminescence dosimeters (TLDs) were used to compare the results. Results : No significant difference was found in the detector sensitivity for static and rotational procedures. The average detector sensitivity for static procedures was 1.10 mV/cGy (SD 0.02) while it was 1.12 mV/cGy (SD 0.02) for rotational procedures. The average detector sensitivity found was the same within the experimental uncertainty for static and rotational dose deliveries. The MOSFET reading was consistent and its reproducibility was excellent (+0.5%) while there was no significant dependence of field size. The angular dependence of less than 1.0% was observed. There was negligible fading effect of the MOSFET. The MOSFET response was found independent of temperature in the range 18\ub0-30\ub0. The ion chamber readings were assumed to be a reference for the estimation of the MOSFET calibration factor. The ion chamber and the TLD were in good agreement (+2%) with each other. Conclusion : This study deals only with the measurements and calibration performed on the surface of the phantom. MOSFET was calibrated and validated for phantom surface measurements for a 6-MV photon beam generated by a tomotherapy machine. The sensitivity of the detector was the same for both modes of treatment delivery with tomotherapy. The performance of the MOSFET was validated for and satisfactory for the helical tomotherapy Hi-Art II unit. However, MOSFET may be used for in vivo surface dosimetry only after it is calibrated under the conditions replicating as much as possible the manner in which the dosimeter will be used clinically

On the transit dose from motorized wedge treatment in Equinox-80 telecobalt unit

Purpose: To estimate the transit dose from motorized wedge (MW) treatment in Equinox-80 telecobalt machine. Materials and Methods: Two plans were generated in Eclipse treatment planning system with universal wedge (UW) and MW each for 10x10 cm 2 . The transit dose was measured with 0.6 cc cylindrical ion chamber and thermoluminescent dosimeters (TLD) chips at a depth of 5 cm with source to axis distance (SAD) 80 cm. Results: The measured dose with ion chamber was in well agreement with the calculated dose from Eclipse within ± 2%. The planned dose was 100 cGy while the measured absorbed dose with ion chamber for 15°, 30°, 45° and 60° MW treatment was found to be 100.94, 101.04, 100.72 and 99.33 cGy respectively. For 15°, 30°, 45° and 60° UW treatment, the measured absorbed dose was 99.33, 97.67, 97.77 and 99.57 cGy respectively. Similarly the measured absorbed dose with TLD was within ± 3% with the planned dose for universal wedge (UW) and MW. From the experimental measurements, it was found that there was no significant contribution of transit dose during MW treatment. Conclusion: The actual measurements carried out with ion chamber in Equinox-80 machine for UW and MW revealed no variation between the doses delivered. The doses were comparable for both UW and MW treatments. The results from TLD measurements additionally confirmed no variation between the doses delivered with UW and MW. It was also demonstrated that the observed excess or less transit dose with MW does not have any significant clinical impact. This assured the safe dose delivery with MW

On the transit dose from motorized wedge treatment in Equinox-80 telecobalt unit

Purpose: To estimate the transit dose from motorized wedge (MW) treatment in Equinox-80 telecobalt machine. Materials and Methods: Two plans were generated in Eclipse treatment planning system with universal wedge (UW) and MW each for 10x10 cm 2 . The transit dose was measured with 0.6 cc cylindrical ion chamber and thermoluminescent dosimeters (TLD) chips at a depth of 5 cm with source to axis distance (SAD) 80 cm. Results: The measured dose with ion chamber was in well agreement with the calculated dose from Eclipse within \ub1 2%. The planned dose was 100 cGy while the measured absorbed dose with ion chamber for 15\ub0, 30\ub0, 45\ub0 and 60\ub0 MW treatment was found to be 100.94, 101.04, 100.72 and 99.33 cGy respectively. For 15\ub0, 30\ub0, 45\ub0 and 60\ub0 UW treatment, the measured absorbed dose was 99.33, 97.67, 97.77 and 99.57 cGy respectively. Similarly the measured absorbed dose with TLD was within \ub1 3% with the planned dose for universal wedge (UW) and MW. From the experimental measurements, it was found that there was no significant contribution of transit dose during MW treatment. Conclusion: The actual measurements carried out with ion chamber in Equinox-80 machine for UW and MW revealed no variation between the doses delivered. The doses were comparable for both UW and MW treatments. The results from TLD measurements additionally confirmed no variation between the doses delivered with UW and MW. It was also demonstrated that the observed excess or less transit dose with MW does not have any significant clinical impact. This assured the safe dose delivery with MW