Macros to Quantify Exosome Release and Autophagy at the Neuromuscular Junction of Drosophila Melanogaster

Automatic quantification of image parameters is a powerful and necessary tool to explore and analyze crucial cell biological processes. This article describes two ImageJ/Fiji automated macros to approach the analysis of synaptic autophagy and exosome release from 2D confocal images. Emerging studies point out that exosome biogenesis and autophagy share molecular and organelle components. Indeed, the crosstalk between these two processes may be relevant for brain physiology, neuronal development, and the onset/progression of neurodegenerative disorders. In this context, we describe here the macros “Autophagoquant” and “Exoquant” to assess the quantification of autophagosomes and exosomes at the neuronal presynapse of the Neuromuscular Junction (NMJ) in Drosophila melanogaster using confocal microscopy images. The Drosophila NMJ is a valuable model for the study of synapse biology, autophagy, and exosome release. By use of Autophagoquant and Exoquant, researchers can have an unbiased, standardized, and rapid tool to analyze autophagy and exosomal release in Drosophila NMJ.Code available at: https://github.com/IreneSaMi/Exoquant-Autophagoquant

A Tool for Rapid Assessment of Functional Outcomes in Patients with Head and Neck Cancer

Head and neck cancer (HNC) and its treatment can lead to various functional impairments. We developed and validated an instrument for rapid physician-rated assessment of basic functional outcomes in HNC patients. HNC-relevant functional domains were identified through a literature review and assigned to verbal ratings based on observable criteria. The instrument draft was subjected to systematic expert review to assess its face and content validity. Finally, the empirical validity, reliability, and responsiveness of the expert-adapted Functional Integrity in Head and Neck Cancer (HNC-FIT) scales were assessed in healthy controls and in HNC patients. A matrix of the 6 functional domains of oral food intake, respiration, speech, pain, mood, and neck and shoulder mobility was created, each with 5 verbal rating levels. Face and content validity levels of the HNC-FIT scales were judged to be adequate by 17 experts. In 37 control subjects, 24 patients with HNC before treatment, and in 60 HNC patients after treatment, the HNC-FIT ratings in the 3 groups behaved as expected and functional domains correlated closely with the outcome of corresponding scales of the EORTC-HN35-QoL questionnaire, indicating good construct and criterion validity. Interrater reliability (rICC) was ≥0.9 for all functional domains and retest reliability (rICC) was ≥0.93 for all domains except mood (rICC = 0.71). The treatment effect size (eta-square) as a measure of responsiveness was ≥0.15 (p < 0.01) for fall domains except for breathing and neck and shoulder mobility. The median HNC-FIT scale completion time was 1 min 17 s. The HNC-FIT scale is a rapid tool for physician-rated assessment of functional outcomes in HNC patients with good validity, reliability, and responsiveness.

Routine Monitoring of Instrument Stability in a Milk Testing Laboratory With ASCA: A Pilot Study

Mid-infrared spectroscopy has been developed as a reliable and rapid tool for routine analysis of fat, protein, lactose and other components in liquid milk. However, variations within and between FTIR instruments, even within the same milk testing laboratory, present a challenge to the accuracy of measurement of particularly minor components in the milk, such as individual fatty acids or proteins. In this study we have used Analysis of variance–Simultaneous Component Analysis (ASCA), to monitor the spectral variation between and within each of four different FOSS FTIR spectrometers over each week in an independent milk testing laboratory over 4 years, between August 2017 and March 2021 (223 weeks). On everyday of each week, spectra of the same pilot milk sample were recorded approximately every hour on each of the four instruments. Overall, variations between instruments had the largest effect on spectral variation over each week, making a significant contribution every week. Within each instrument, day-to-day variations over the week were also significant for all but two of the weeks measured, however it contributed less to the variance overall. At certain times other factors not explained by weekday variation or inter-instrument variation dominated the variance in the spectra. Examination of the scores and loadings of the weekly ASCA analysis allowed identification of changes in the spectral regions affected by drifts in each instrument over time. This was found to particularly affect some of the fatty acid predictions.

Experimental Research on Deep-And-Narrow Micromilled Grooves Using a Self-Fabricated PCD Micro-Cutter

Deep-and-narrow micro-grooves are the common functional structures of miniature parts. The fabrication of the micromilled grooves with high quality and accuracy is the essential guarantee of the causative performance for these miniature parts, and micromilling is the most versatile process to machine such micro-grooves. However, micromilling technology is a highly tool-dependent process, and the commercial carbide micromilling cutter has shown obvious deficiencies in terms of rapid tool wear and inferior machined quality during the machining process. In this paper, a polycrystalline diamond (PCD) micromilling cutter with a large-aspect-ratio (LAR) was designed and prepared by the self-proposed hybrid fabrication method of laser and precision grinding. Micromilling experiments on oxygen-free copper were conducted, and the carbide micromilling cutter was selected in the comparative experiments. The variations of milling forces and specific energy were analyzed through the parameter experiments. Then, the surface quality, machined accuracy and tool wear were further investigated. Results showed that the PCD micromilling cutter with an aspect ratio of 3.25 was successfully manufactured by the proposed hybrid method. The self-fabricated PCD micromilling cutter presented remarkable superiority in terms of the surface quality, machined accuracy, and tool wear when preparing deep-and-narrow micro-grooves. Finally, a satisfactory micromilled groove with an aspect ratio of 2.5 was achieved with the self-fabricated LAR PCD cutter under the optimized conditions.

Sugarcane smut, caused by Sporisorium scitamineum, a major disease of sugarcane – a contemporary review

Sugarcane smut caused by the fungus Sporisorium scitamineum is one of the major diseases of sugarcane worldwide, causing significant losses in productivity and profitability of this perennial crop. Teliospores of this fungus are air-borne, can travel long distances and remain viable in hot and dry conditions for more than six months. The disease is easily recognised by its long ‘whip’-like sorus produced on the apex or side shoots of sugarcane stalks. Each sorus can release up to 100 million teliospores in a day; the spores are relatively small (≤7.5 µ), light and can survive in harsh environmental conditions. The air-borne teliospores are the primary mode of smut spread around the world and across cane-growing regions. The most effective method of managing this disease is via resistant varieties. Due to the complex genomic makeup of sugarcane, selection for resistant traits is difficult in sugarcane breeding programs. In recent times, the application of molecular markers as a rapid tool of discarding susceptible genotypes early in the selection program has been investigated. Large effect resistance loci have been identified and have the potential to be utilised for marker-assisted selection to increase the frequency of resistant breeding lines in breeding programs. Recent developments in “omics” technologies (genomics, transcriptomics, proteomics, and metabolomics) have contributed to our understanding and provided insights into the mechanism of resistance and susceptibility. This knowledge will further our understanding of smut and its interactions with sugarcane genotypes, and aid in the development of durable resistant varieties.

Check-All-That-Apply (CATA) Test to Investigate the Consumers’ Perception of Olive Oil Sensory Properties: Effect of Storage Time and Packaging Material

Sensory changes during shelf-life of oils have been mostly studied by descriptive methods, while consumer-based approaches have been poorly explored. This study assessed the variations in consumers’ liking and sensory perception of extra virgin olive oil (EVOO) and olive oil (OO) packaged in glass, polyethylene terephthalate and tinplate. After 2, 10 and 19 months of storage, oil perception was investigated with consumers (n = 50) performing both a liking test and a check-all-that-apply (CATA) test. No significant effect of the packaging material on consumers’ response was found, whereas storage time negatively affected the sensory properties of and acceptability of OOs and EVOOs from the 10th month of storage. The CATA test results revealed the sensory changes in oils over 19 months, mainly described as a decrease in pungency for EVOO and a decrease in herbaceous and ripe fruitiness in OO. The CATA technique combined with the liking test allowed the drivers of liking (“olive” for OO and “green fruitiness” for EVOO) and disliking (“bitter” and “pungent” for EVOO) to be identified. In conclusion, the sensory approach based both on CATA technique and liking test seems promising as a rapid tool to evaluate the changes in sensory properties perceivable during the shelf-life of oils.