Retrospective analysis of random and systematic errors in radiation therapy of head and neck cancer patients and its clinical predictive implications with VMAT treatment

Background: The accuracy of radiotherapy is based on the matching of 2D portal/CBCT image with a reference image. The aim of this study is to determine the random and systematic setup errors (in cm) in radiotherapy of head and neck cancer patients and to derive the setup margin and its clinical implications.Methods: Author retrospectively reviewed the records of 25 head and neck cancer (HNC) patients treated with radiotherapy between Dec 2017 and July 2018. After immobilization, setup accuracy was assessed by registration of XVI image with planning reference image using Elekta XVI image guidance system and the isocenter correction was applied. For each patient 10 CBCT image sets were taken. The translational errors in X, Y and Z directions were used to estimate systematic (Σ) and random (σ) errors and to derive the final setup margin by using van Herk’s formula (2.5Σ + 0.7σ).Results: The mean translational errors ranges from -0.23 cm to 0.32 cm in Lateral (X), -0.15 to 0.16 cm in Longitudinal (Y) and -0.11 to 0.17 cm in vertical (Z) directions. The Mean and SD for systematic errors 0.21±0.13, 0.11±0.18, 0.14±0.11 and random error (in cm) are -0.03±0.33, 0.00±0.21 and 0.05±0.30 in X, Y and Z axis respectively. The final total margin for CTV to PTV including setup margin in the X, Y and Z directions (in cm) were 0.56, 0.61, and 0.47 respectively.Conclusion: Thus, the precise immobilization techniques are very important to reduce the setup margins, and the number of CBCTs during head and neck radiotherapy treatment

Quantifying the infectiousness of post-kala-azar dermal leishmaniasis towards sandflies

Background In the Indian subcontinent, visceral leishmaniasis (VL) incidence is on track to reach elimination goals by 2020 in nearly all endemic districts. Although not included in official targets, previous data suggest post-kala-azar dermal leishmaniasis (PKDL) patients can act as an infection reservoir. Methods We conducted xenodiagnosis on 47 PKDL patients and 15 VL patients using laboratory-reared Phlebotomus argentipes. In direct xenodiagnosis, flies were allowed to feed on the patient’s skin for 15 minutes. For indirect xenodiagnosis, flies were fed through a membrane on the patient’s blood. Five days later, blood-fed flies were dissected and examined by microscopy and/or PCR. A 3-mm skin snip biopsy (PKDL) or venous blood VL) was processed by quantitative PCR. Results Twenty-seven PKDL patients (57.4%) had positive results by direct and/or indirect xenodiagnosis. Direct was significantly more sensitive than indirect xenodiagnosis (55.3% vs 6.4%, p 1 log10 unit higher than those with negative results (2.88 vs 1.66, p<0.0001). In a multivariable model, parasite load, nodular lesions and positive skin microscopy were significantly associated with positive xenodiagnosis. Blood parasite load was the strongest predictor for VL. Compared to VL, nodular PKDL was more likely and macular PKDL less likely to result in positive xenodiagnosis, but neither difference reached statistical significance. Conclusions Nodular and macular PKDL, and VL, can be infectious to sand flies. Active PKDL case detection and prompt treatment should be instituted and maintained as an integral part of VL control and elimination programs

Validation of the COSMIC Radio Occultation Data over Gadanki (13.48°N, 79.2°E): A Tropical Region

Constellation Observing System for Meteorology Ionosphere and Climate (COSMIC), consisting of six Low Earth Orbit (LEO) Global Position System (GPS) receivers, on board the Formosat Satellite 3 (FORMOSAT-3) is providing dense observations of density, refractivity, temperature and water vapor profiles of the neutral atmosphere since middle of July 2006. Special radiosonde (Väisälä) campaign was conducted at Gadanki (13.48°N, 79.18°E), a tropical site in India, during July 2006 to March 2007 to validate these meteorological parameters. Co-located Nd: YAG Rayleigh lidar was also operated during the overpass of COSMIC and is utilized to validate the temperatures in the height range of 30 to 40 km. Atotal of 142 overpasses occurred during the above mentioned period within 300 km distance from Gadanki out of which 41 overpasses occurred within a time difference of ±4 hours of radiosonde launch. In addition, 18 overpasses occurred within the time difference of ±4 hours of lidar operation. A detailed comparison has been made with all these overpasses for the refractivity, temperature and water vapor obtained from COSMIC. The water vapor comparison has shown generally a good agreement with a mean difference of 5 - 10% below 6 - 7 km. Although there is a colder bias between COSMIC and radiosonde, a very good comparison in temperature is also found between 10 and 27 km with a mean difference of less than 1 K (RMS difference is only 0.64 K). There exists a large difference in temperature of about 8 K between 30 and 40 km (between COSMIC and lidar). Possible reasons for these large differences are given. There was one event that occurred just over Gadanki for which a detailed comparison has been made with special emphasis on water vapor retrievals. Sensitivity test is also done on the fractional difference in N for the event that occurred on 24 July 2006 between COSMIC (1D-var) and radiosonde and found that pressure plays a key role than temperature in determining the refractivity

Reflections on the CLIVAR Early Career Scientists Symposium 2016

We present a summary report of the CLIVAR Early Career Scientists Symposium, a three-day event associated with the CLIVAR Open Science Conference held in Qingdao, China during September 2016. The Symposium aimed to capture the ideas of early career researchers on pressing science priorities, imminent challenges, and emerging opportunities to help guide the future evolution of CLIVAR. We identified the need for improving process-based understanding and predictability of regional climate variability and change, moving toward seamless predictions, and improving and expanding global observations. We emphasize the need for increasingly open science, including universal access to data, code, and publications as well as opportunities for international cooperation and exchange. As the next generation of climate scientists, we are dedicated to overcome the challenges outlined in this summary and are looking forward to advancing CLIVAR???s mission and activities

Evaluation of Rapid Extraction Methods Coupled with a Recombinase Polymerase Amplification Assay for Point-of-Need Diagnosis of Post-Kala-Azar Dermal Leishmaniasis

To detect Post-kala-azar leishmaniasis (PKDL) cases, several molecular methods with promising diagnostic efficacy have been developed that involve complicated and expensive DNA extraction methods, thus limiting their application in resource-poor settings. As an alternative, we evaluated two rapid DNA extraction methods and determined their impact on the detection of the parasite DNA using our newly developed recombinase polymerase amplification (RPA) assay. Skin samples were collected from suspected PKDL cases following their diagnosis through national guidelines. The extracted DNA from three skin biopsy samples using three different extraction methods was subjected to RPA and qPCR. The qPCR and RPA assays exhibited highest sensitivities when reference DNA extraction method using Qiagen (Q) kit was followed. In contrast, the sensitivity of the RPA assay dropped to 76.7% and 63.3%, respectively, when the boil & spin (B&S) and SpeedXtract (SE) rapid extraction methods were performed. Despite this compromised sensitivity, the B&S-RPA technique yielded an excellent agreement with both Q-qPCR (k = 0.828) and Q-RPA (k = 0.831) techniques. As expected, the reference DNA extraction method was found to be superior in terms of diagnostic efficacy. Finally, to apply the rapid DNA extraction methods in resource-constrained settings, further methodological refinement is warranted to improve DNA yield and purity through rigorous experiments

Evaluation of Loopamp™ Leishmania Detection Kit and Leishmania Antigen ELISA for Post-Elimination Detection and Management of Visceral Leishmaniasis in Bangladesh

With reduced prevalence of visceral leishmaniasis (VL) in the Indian subcontinent (ISC), direct and field deployable diagnostic tests are needed to implement an effective diagnostic and surveillance algorithm for post-elimination VL control. In this regard, here we investigated the diagnostic efficacies of a loop-mediated isothermal amplification (LAMP) assay (Loopamp™ Leishmania Detection Kit, Eiken Chemical CO., Ltd, Japan), a real-time quantitative PCR assay (qPCR) and the Leishmania antigen ELISA (CLIN-TECH, UK) with different sampling techniques and evaluated their prospect to incorporate into post-elimination VL control strategies. Eighty clinically and rK39 rapid diagnostic test confirmed VL cases and 80 endemic healthy controls were enrolled in the study. Peripheral blood and dried blood spots (DBS) were collected from all the participants at the time of diagnosis. DNA was extracted from whole blood (WB) and DBS via silica columns (QIAGEN) and boil & spin (B&S) methods and tested with qPCR and Loopamp. Urine was collected from all participants at the time of diagnosis and was directly subjected to the Leishmania antigen ELISA. 41 patients were followed up and urine samples were collected at day 30 and day 180 after treatment and ELISA was performed. The sensitivities of the Loopamp-WB(B&S) and Loopamp-WB(QIA) were 96.2% (95% CI 89·43-99·22) and 95% (95% CI 87·69-98·62) respectively. The sensitivity of Loopamp- DBS(QIA) was 85% (95% CI 75·26- 92·00). The sensitivities of the qPCR-WB(QIA) and qPCR-DBS(QIA) were 93.8% (95% CI 86·01-97·94) and 72.5% (95% CI 61·38-81·90) respectively. The specificity of all molecular assays was 100%. The sensitivity and specificity of the Leishmania antigen ELISA were 97.5% (95% CI 91·47-99·70) and 91.95% (95% CI 84·12-96·70) respectively. The Leishmania antigen ELISA depicted clinical cure at day 180 in all the followed-up cases. Efficacy and sustainability identify the Loopamp-WB(B&S) and the Leishmania antigen ELISA as promising and minimally invasive VL diagnostic tools to support VL diagnostic and surveillance activities respectively in the post-elimination era

Evaluation of Loopamp™ Leishmania Detection Kit and Leishmania Antigen ELISA for Post-Elimination Detection and Management of Visceral Leishmaniasis in Bangladesh

With reduced prevalence of visceral leishmaniasis (VL) in the Indian subcontinent (ISC), direct and field deployable diagnostic tests are needed to implement an effective diagnostic and surveillance algorithm for post-elimination VL control. In this regard, here we investigated the diagnostic efficacies of a loop-mediated isothermal amplification (LAMP) assay (Loopamp™ Leishmania Detection Kit, Eiken Chemical CO., Ltd, Japan), a real-time quantitative PCR assay (qPCR) and the Leishmania antigen ELISA (CLIN-TECH, UK) with different sampling techniques and evaluated their prospect to incorporate into post-elimination VL control strategies. Eighty clinically and rK39 rapid diagnostic test confirmed VL cases and 80 endemic healthy controls were enrolled in the study. Peripheral blood and dried blood spots (DBS) were collected from all the participants at the time of diagnosis. DNA was extracted from whole blood (WB) and DBS via silica columns (QIAGEN) and boil & spin (B&S) methods and tested with qPCR and Loopamp. Urine was collected from all participants at the time of diagnosis and was directly subjected to the Leishmania antigen ELISA. 41 patients were followed up and urine samples were collected at day 30 and day 180 after treatment and ELISA was performed. The sensitivities of the Loopamp-WB(B&S) and Loopamp-WB(QIA) were 96.2% (95% CI 89·43-99·22) and 95% (95% CI 87·69-98·62) respectively. The sensitivity of Loopamp-DBS(QIA) was 85% (95% CI 75·26- 92·00). The sensitivities of the qPCR-WB(QIA) and qPCR-DBS(QIA) were 93.8% (95% CI 86·01-97·94) and 72.5% (95% CI 61·38-81·90) respectively. The specificity of all molecular assays was 100%. The sensitivity and specificity of the Leishmania antigen ELISA were 97.5% (95% CI 91·47-99·70) and 91.95% (95% CI 84·12-96·70) respectively. The Leishmania antigen ELISA depicted clinical cure at day 180 in all the followed-up cases. Efficacy and sustainability identify the Loopamp-WB(B&S) and the Leishmania antigen ELISA as promising and minimally invasive VL diagnostic tools to support VL diagnostic and surveillance activities respectively in the post-elimination era

Gauging the skin resident Leishmania parasites through a loop mediated isothermal amplification (LAMP) assay in post-kala-azar dermal leishmaniasis

Despite the availability of highly sensitive polymerase chain reaction (PCR)-based methods, the dearth of remotely deployable diagnostic tools circumvents the early and accurate detection of individuals with post-kala-azar dermal leishmaniasis (PKDL). Here, we evaluate a design-locked loop-mediated isothermal amplification (LAMP) assay to diagnose PKDL. A total of 76 snip-skin samples collected from individuals with probable PKDL (clinical presentation and a positive rK39 rapid diagnostic test (RDT)) were assessed by microscopy, qPCR, and LAMP. An equal number of age and sex-matched healthy controls were included to determine the specificity of the LAMP assay. The LAMP assay with a Qiagen DNA extraction (Q-LAMP) showed a promising sensitivity of 72.37% (95% CI: 60.91–82.01%) for identifying the PKDL cases. LAMP assay sensitivity declined when the DNA was extracted using a boil-spin method. Q-qPCR showed 68.42% (56.75–78.61%) sensitivity, comparable to LAMP and with an excellent agreement, whereas the microscopy exhibited a weak sensitivity of 39.47% (28.44–51.35%). When microscopy and/or qPCR were considered the gold standard, Q-LAMP exhibited an elevated sensitivity of 89.7% (95% CI: 78.83–96.11%) for detection of PKDL cases and Bayesian latent class modeling substantiated the excellent sensitivity of the assay. All healthy controls were found to be negative. Notwithstanding the optimum efficiency of the LAMP assay towards the detection of PKDL cases, further optimization of the boil-spin method is warranted to permit remote use of the assay