Integrated approach to functional analysis of an ERBB2 variant of unknown significance detected by a cancer gene panel test

Purpose Dealing with variants of unknown significance (VUS) is an important issue in the clinical application of NGS-based cancer gene panel tests. We detected a novel ERBB2 extracellular domain VUS, c.1157A > G p.(E401G), in a cancer gene panel test. Since the mechanisms of activation by ERBB2 extracellular domain (ECD) variants are not fully understood, we aimed to clarify those mechanisms and the biological functions of ERBB2 E401G. Methods ERBB2 E401G was selected as VUS for analysis because multiple software tools predicted its pathogenicity. We prepared ERBB2 expression vectors with the E401G variant as well as vectors with S310F and E321G, which are known to be activating mutations. On the basis of wild-type ERBB2 or mutant ERBB2 expression in cell lines without ERBB2 amplification or variants, we evaluated the phosphorylation of human epidermal growth factor receptor 2 and related proteins, and investigated with molecular dynamics (MD) simulation the mechanisms conferred by the variants. The biological effects of ERBB2 E401G were also investigated, both in vitro and in vivo. Results We found that ERBB2 E401G enhances C-terminal phosphorylation in a way similar to S310F. MD simulation analysis revealed that these variants maintain the stability of the EGFR-HER2 heterodimer in a ligand-independent manner. Moreover, ERBB2 E401G-transduced cells showed an increased invasive capacity in vitro and an increased tumor growth capacity in vivo. Conclusion Our results provide important information on the activating mechanisms of ERBB2 extracellular domain (ECD) variants and illustrate a model workflow integrating wet and dry bench processes for the analysis of VUS detected with cancer gene panel tests.

Exploitation of Yeasts with Probiotic Traits for Kefir Production: Effectiveness of the Microbial Consortium

Kefir is a fermented milk made by beneficial lactic acid bacteria and yeasts inoculated as grains or free cultures. In this work, five yeast strains with probiotic aptitudes belonging to Candida zeylanoides, Yarrowia lipolytica, Kluyveromyces lactis, and Debaryomyces hansenii species were assessed in a defined consortium, in co-culture with a commercial strain of Lactobacillus casei, in order to evaluate the yeasts’ fermentation performance during kefir production, using different milks. The concentration of each yeast was modulated to obtain a stable consortium that was not negatively affected by the bacteria. Furthermore, all yeasts remained viable for five weeks at 4 °C, reaching about 8.00 Log CFU in 150 mL of kefir, a volume corresponding to a pot of a commercial product. The yeasts consortium showed a suitable fermentation performance in all milks, conferring peculiar and distinctive analytical and aromatic properties to the kefirs, confirmed by a pleasant taste. Overall, the panel test revealed that the cow’s and sheep’s kefir were more appreciated than the others; this evaluation was supported by a distinctive fermentation by-products’ content that positively influences the final aroma, conferring to the kefir exalted taste and complexity. These results allow us to propose the yeasts consortium as a versatile and promising multistarter candidate able to affect industrial kefir with both recognizable organoleptic properties and probiotic aptitudes.

Drenching Bovine Colostrum, Quercetin or Fructo-Oligosaccharides Has No Effect on Health or Survival of Low Birth Weight Piglets

The introduction of hyperprolific sows has resulted in more low birth weight (LBW) piglets, accompanied by higher mortality. A possible strategy to enhance the resilience and survival of LBW piglets is oral supplementation (drenching) of bioactive substances. This study evaluated the supplementation of bovine colostrum, short-chain fructo-oligosaccharides (scFOS) or quercetin that were dissolved separately in a milk replacer. The study was divided into two sub-experiments. First, the milk replacer was compared with a sham drenched group. Secondly, each dissolved compound was compared with the milk replacer. The LBW piglets, defined as weighing between (mean litter birth weight −1*SD) and (mean litter birth weight −2.5*SD), were randomly allocated to the different treatments and drenched once a day for seven days. On day 1, 3, 9, 24 and 38, piglets were weighed and scored for skin lesions. Blood samples were collected on day 9 and 38 and analyzed to determine glucose, non-esterified fatty acids, urea, immunoglobulin G, insulin-like growth factor 1, and a standard blood panel test. There was no difference between sham drenched piglets and piglets that were drenched with milk replacer regarding any of the parameters. No effect was observed between the milk replacer group and any of the bioactive compounds either, except a higher mortality within the scFOS group. In conclusion, this study showed that drenching the evaluated bioactive compounds, in the used dosages, did not improve LBW piglets’ resilience or survival and more research is required to determine the effect of scFOS on small piglets.

Design of a radiant panel test at reduced scale for the high throughput development of artificial turf structures

Artificial turf structures are increasingly used in closed areas and have to comply with the European fire standard for building products (EN ISO 13501-1). The main test to evaluate the fire performance of flooring products is the EN ISO 9239-1 radiant panel test. The test principle is to determine the critical heat flux of floorings exposed to a forced ignition and a specific heat flux profile. As large amounts of material are needed to perform the test, the development of a radiant panel test at reduced scale was considered. The experimental design methodology was implemented to mimic the heat flux profile. The fire performance of artificial turf structures was evaluated at both scales and the results were compared. The burnt lengths of the specimens and thus the critical heat flux are similar for both scales. Thus, the downscaled device could advantageously be used for high throughput development of artificial turf structures.

ACT-8 Current status and prospects for the treatment of malignant glioma using cancer gene panel tests

Introduction: The cancer gene panel test was covered by insurance in June 2019. Our institution started the test in May 2020 and has experienced 10 cases, so we will report on the current status and future prospects. Methods: The subjects were 10 patients who underwent the cancer gene panel test using FoundationOne CDx. Results:The cases included 8 glioblastomas, an anaplastic astrocytoma, and an anaplastic oligodendroglioma. The total number of tumor mutational burden (TMB) was judged to be low in all cases, and the microsatellite instability test (MSI) showed no instability in all cases (MSI-Stable). The total number of genetic changes detected was 11 ± 5.0, oncogene mutations were 5.3 ± 2.4, and gene mutations of unknown relevance to cancer were 5.7 ± 2.8. Major oncogene mutations were IDH1 mutation in 4 cases, ATRX mutation in 2 cases, TP53 mutation in 6 cases, and BRAF V600E mutation in 1 case. Based on the test results, a 25-year-old man with BRAF V600E mutation was initiated into the NCCH1901 study (Patient-Proposed Healthcare Services). A case with IDH1 mutation (47-year-old male) entered a phase I clinical trial of a mutant IDH1 inhibitor. It is estimated that the chance of finding an appropriate drug by cancer gene panel test is about 10–20%. However, in cases that are resistant to standard treatment, the benefits can be expected if the drugs associated with the cancer gene panel test can be used. Conclusions: Although Malignant gliomas are often TMB-low and MSI-stable and the response rate to molecular-targeted drugs and other therapies is not high, there are some cases that can be salvaged by performing the cancer gene panel test. It is suggested that the active use of cancer gene panel test may contribute to the development of new drugs with high response rates and the improvement of prognosis.

Formulation of a Sugar-Free Cereal-Based Bar with Improved Nutritional Value: Biological and Sensory Analysis

Cereal bars are eaten regularly due to their nutritive values and ease of consumption. Furthermore, new cereal bars were recently introduced to serve the growing market of functional food. In this work, we aimed at developing an innovative sugar-free cereal bar with high protein and fibre content, showing functional effects and with improved sensory properties over cognate commercial bars. We developed an innovative recipe by substituting honey with a xanthan gum/erythritol mixture as binder, and by manufacturing bars in a conventional industrial apparatus. The new formula was characterised by high content of proteins (27.7%) and fibres (8.7%), whereas carbohydrates resulted drastically reduced (26.7%). Panel test analysis showed that bars hold satisfactory sensory properties (aroma, flavour and taste), whereas some texture parameters were positively changed, as hardness and adhesiveness were significantly reduced. In vitro challenge experiments showed that the phenolic components isolated from the bar matrix significantly increased the levels of glutathione and glutathione reductase in Caco-2 cells, a model of enterocytes (P<0.05). In conclusion, we produced a sugar-free cereal bar on an industrial scale with antioxidant properties, and with improved nutritional and texture parameters in comparison with a conventional product.

Research on Real-time Sensing Technology of Aerospace Interactive Behavior Based on Infrared Array Force Tactile

Aerospace robots can replace astronauts to complete dangerous work in the expanded space. How to efficiently and reliably interact with robots in complex and harsh space environment has become an urgent problem and technical challenge. Based on the infrared array force tactile technology, an infrared point force touch detection system is developed to realize the intention sensing of aerospace human-robot interaction. Based on multipoint positioning technology combined with image method and global combined optimal tracking method, the control software is designed, and the effective human-robot interaction in the process of cooperation is realized. Finally, the touch panel test effect of the system is given. This research can provide a theoretical basis for the practical application of aerospace human-robot interaction.

Impact of immunological, hematological and biochemical markers on discordant partners of Hepatitis B infection in Enugu State, Eastern Nigeria

Globally, hepatitis B infection is a serious health problem as its morbities are difficult to handle with the commonly used antivirals. This study therefore aims to evaluate the serological, biochemical and hematological markers of Hepatitis B virus infection in a discordant partner in an attempt to aid proper diagnosis, management and control of the infection. This is a cross-sectional study consisting of 150 subjects recruited using a simple random sampling technique. HBsAg was screened using a rapid ELISA diagnostic strip and rescreened later for confirmation using Ichroma Fluorescence Immunoassay (FIA). HBV Serological markers were determined using HBV 5 panel test, hematological parameters were assessed using Mindray BC 10 automated counter, while liver enzymes were estimated using COBAS 111 analyzer. Statistical analysis was performed using Graph Pad Prism. This study demonstrated a varying percentage detection rates of HBV serological markers in both groups: Discordant and Concordant partners (HBsAg- 62%, HBsAb- 4.2%, HBeAg- 2.5%, HBeAb- 50%, HBcAb-60.8%), and Control partners(HBsAg-0%, HBsAb-23.3%, HBeAg-0%, HBeAb- 0%,and HBcAb- 6.7%). The liver enzymes showed significant mean values (P<0.001) for both ALT and AST positive partners when compared to their negative counterparts. Hematological parameters, only hemoglobulin showed a significant mean (P<0.001) on male subjects as against females, while white blood cell and platelets were not statistically significant. There is a need for hepatitis B panel test inclusion in the routine investigation for Hepatitis B viral infection which will contribute immensely in proper clinical management and control of the infection in conjunction with vaccination, while periodic assessment of liver enzymes will ensure proper management of chronic hepatitis B infections.

Comparison of the Chemical and Sensorial Evaluation of Dark Chocolate Bars

As it mimics olfactory perception, headspace analysis is frequently used for examination of products like chocolate, in which aroma is a key feature. Chemical analysis by itself, however, only provides half the picture, as final consumer’s perception cannot be compared to that of a Gas Chromatography-Mass Spectrometry (GC-MS) port, but rather to a panel test assessment. The aim of the present study was the evaluation of combined chemical (by means of headspace solid-phase microextraction and GC-MS) and panel test data (by means of a sensory evaluation operated by 6 untrained panelists) obtained for 24 dark chocolate bars to assess whether these can discriminate between bars from different brands belonging to different commercial segments (hard discount, HD; supermarket, SM; organic bars, BIO). In all samples, with the only exception of one supermarket bar (in which esters exhibited the highest relative abundance), pyrazines were detected as the most abundant chemical class (HD: 56.3–74.2%; BIO: 52.0–76.4%; SM: 31.2–88.9%). Non-terpene alcohols, aldehydes, and esters followed as quantitatively relevant groups of compounds. The obtained data was then subjected to hierarchical cluster (HCA) and principal component (PCA) analysis. The statistical distribution of samples obtained for the chemical data did not match that obtained with panelists’ sensorial data. Moreover, although an overall ability of grouping samples of the same commercial origin was evidenced for hard discount and supermarket bars, no sharp grouping was possible.