Efficacy of Eicosapentaenoic Acid (EPA) Containing Protein Supplement in Preventing Weight Loss in Head and Neck Cancer Patients Undergoing Curative Radiotherapy: Retrospective Observations with Historical Controls.

Weight loss is a major issue in Head and Neck cancer (HNC) patients undergoing curative radiotherapy. The principal objective of the study was to observe whether eicosapentaenoic acid (EPA) containing protein supplement was effective in mitigating the weight loss during in hospitalized HNC undergoing curative radiotherapy. A retrospective study was performed based on clinical, treatment and nutritional data of 53 patients received EPA containing supplement during their curative radiotherapy from October 2014 to January 2015 and was compared with 88 historical control group of patients (October 2013 to June 2014) who had indigenous protein rich diet planned by the dietician in the period immediately before the implementation of providing EPA containing protein supplement to the patients. The data was stratified based on gender, age, weight, treatment modalities, stage and site of cancer and analysed using unpaired t test. A p value of < 0.05 was considered significant. The results indicate there was no significant difference in the patient tumor and clinical details. The results indicate that the percent change in weight loss was less in the EPA cohorts when calculated from weight (P < 0.006) and Body Mass Index (BMI) perspective (P < 0.003). Detail analysis suggested that beneficial effects were more in males (P < 0.01), people affected with oral cancer (P < 0.02), people below the age of 40 (P < 0.001), and in people with early stage cancer (P < 0.003). Cumulatively all these results suggest that administering EPA containing protein supplement was effective in arresting weight loss in HNC patients undergoing curative radiotherapy.

Link between Blood Cell-Associated Inflammatory Indices and Chemotherapy-Induced Hyperglycemia in Women Affected with Breast Cancer: Clinical Studies.

Krishna PrasadBackground Development of treatment-induced hyperglycemia/diabetes is a considerable problem in women undergoing chemotherapy for breast cancer. In this study, baseline levels of blood cell-associated inflammatory indices (BCAII) were analyzed to understand their role in the development of treatment-induced hyperglycemia and diabetogenesis. Materials and Methods This was a retrospective study, and information on women who were normoglycemic and nondiabetic and of women who were diabetic at the beginning of the treatment were collected from files. Demographic, pathology-related details, and complete blood profile were noted. Neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), platelet-to-lymphocyte ratio (PLR), and systemic inflammatory index (SII) which indicate BCAII were calculated. Demographic details were subjected to frequency and percentage, while blood parameters were subjected to one-way analysis of variance followed by post hoc Bonferroni's multiple comparison tests. A p -value of <0.05 was considered significant. Results The results indicated that a significant difference in levels of total count ( p < 0.035), neutrophil, lymphocyte, and platelets ( p < 0.001) were observed. Regarding BCAII, when compared with women who were normoglycemic at the end of treatment, NLR, dNLR, PLR, and SII were significantly high for people who were known diabetics at the beginning of treatment ( p < 0.001). The dNLR ( p = 0.0008), PLR ( p < 0.001), and SII ( p < 0.001) were significant for people who developed secondary hyperglycemia/diabetes, while only dNLR was significant for people who progressed from normal to prediabetes stage ( p = 0.049) Conclusion To the best of the authors' knowledge, this is the first study that indicates difference in baseline BCAII and development of treatment-induced hyperglycemia/diabetes indicating that underlying low levels of inflammation may contribute to diabetogenesis in women affected with breast cancer.

Usefulness of walkathon in improving knowledge on cancer: A cross-sectional study with the participants.

AIM: Globally, walkathon, a type of community event involving walking a determined distance to raise awareness and/or collect donations for a social cause is gaining a lot of public attention. This study was conducted to understand the usefulness of walkathon in creating cancer awareness among the participants. MATERIALS AND METHODS: This was a pre- and post-test study and was carried out on World Cancer Day on February 4, 2019 in Mangalore, Karnataka, India. The study subjects were the walkathon participants willing to be volunteers for the study. Differences in the answers were calculated by ascertaining the difference between the pretest and posttest using McNemar, Chi-square, or Fisher's exact test. A P < 0.05 was considered statistically significant. RESULTS: Pretest scores indicated most volunteers were aware of the role of tobacco and alcohol in cancer, while most were unaware of the importance of fruits and consumption of vegetables and sedentary lifestyle in the development of cancer. Posttest showed a significant increase in knowledge gain (P = 0.003 to <0.001). CONCLUSIONS: For the first time, this study shows Walkathon to be helpful in improving cancer knowledge in the participants. Walkathon is an inexpensive and joyful way to educate people and warrants further study on a range of medical and social themes affecting society to establish its usefulness in public health education.

Evaluating the persistence and stability of a DNA-barcoded microbial system in a mock home environment.

Recent advancements in engineered microbial systems capable of deployment in complex environments have enabled the creation of unique signatures for environmental forensics operations. These microbial systems must be robust, able to thrive in specific environments of interest and contain molecular signatures, enabling the detection of the community across conditions. Furthermore, these systems must balance biocontainment concerns with the stability and persistence required for environmental forensics. Here we evaluate the stability and persistence of a recently described microbial system composed of germination-deficient Bacillus subtilis and Saccharomyces cerevisiae spores containing nonredundant DNA barcodes in a controlled simulated home environment. These spore-based microbial communities were found to be persistent in the simulated environment across 30-day periods and across multiple surface types. To improve the repeatability and reproducibility in detecting the DNA barcodes, we evaluated several spore lysis and sampling processes paired with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) -CRISPR-associated proteins (Cas) detection (Sherlock). Finally, having optimized the detectability of the spores, we demonstrate that we can detect the spores transferring across multiple material types. Together, we further demonstrate the utility of a recently described microbial forensics system and highlight the importance of independent validation and verification of synthetic biology tools and applications. Graphical Abstract.

Variability in cell-free expression reactions can impact qualitative genetic circuit characterization.

Cell-free expression systems provide a suite of tools that are used in applications from sensing to biomanufacturing. One of these applications is genetic circuit prototyping, where the lack of cloning is required and a high degree of control over reaction components and conditions enables rapid testing of design candidates. Many studies have shown utility in the approach for characterizing genetic regulation elements, simple genetic circuit motifs, protein variants or metabolic pathways. However, variability in cell-free expression systems is a known challenge, whether between individuals, laboratories, instruments, or batches of materials. While the issue of variability has begun to be quantified and explored, little effort has been put into understanding the implications of this variability. For genetic circuit prototyping, it is unclear when and how significantly variability in reaction activity will impact qualitative assessments of genetic components, e.g. relative activity between promoters. Here, we explore this question by assessing DNA titrations of seven genetic circuits of increasing complexity using reaction conditions that ostensibly follow the same protocol but vary by person, instrument and material batch. Although the raw activities vary widely between the conditions, by normalizing within each circuit across conditions, reasonably consistent qualitative performance emerges for the simpler circuits. For the most complex case involving expression of three proteins, we observe a departure from this qualitative consistency, offering a provisional cautionary line where normal variability may disrupt reliable reuse of prototyping results. Our results also suggest that a previously described closed loop controller circuit may help to mitigate such variability, encouraging further work to design systems that are robust to variability. Graphical Abstract.

Determining the Binding Kinetics of Peptide Macrocycles Using Bio-Layer Interferometry (BLI).

Bio-Layer Interferometry (BLI) enables the detection and characterization of molecular interactions in real-time without the hassle and interference of labeling. The affinity constant (KD) obtained in the BLI analysis is an excellent indicator of quality of biomolecules such as antibodies, aptamers, peptides, etc. This method was used to analyze a peptide macrocycle against the Abrax protein, but can be used for any peptide macrocycle/analyte system.

Indian Indigenous Fruits as Radioprotective Agents: Past, Present and Future.

Ionising radiation has been an important modality in cancer treatment and its value is immense when surgical intervention is risky or might debilitate/adversely affect the patient. However, the beneficial effect of radiation modality is negated by the damage to the adjacent healthy tissue in the field of radiation. Under these situations, the use of radioprotective compounds that can selectively protect normal tissues against radiation injury is considered very useful. However, research spanning over half a century has shown that there are no ideal radioprotectors available. The United States Food and Drug Administration (FDA or USFDA) approved amifostine, or WR-2721 (Walter Reed-2721) [chemically S-2-(3-aminopropyl-amino) ethyl phosphorothioic acid] is toxic at their optimal concentrations. This has necessitated the need for agents that are safe and easily acceptable to humans. BACKGROUND: Dietary agents with beneficial effects like free radical scavenging, antioxidant and immunomodulatory effects are being recognized as useful and have been investigated for their radioprotective properties. Studies in these lines have shown that the fruits of Aegle marmelos (stone apple or bael), Emblica officinalis or Phyllanthus emblica (Indian gooseberry/amla), Eugenia jambolana or Syzygium jambolana (black plum/jamun), Mangifera indica (mango) and Grewia asiatica (phalsa or falsa) that are originally reported to be indigenous to India have been investigated for their usefulness as radioprotective agents. OBJECTIVE: The objective of this review is to summarize the beneficial effects of the Indian indigenous fruits, stone apple, mango, Indian gooseberry, black plum, and phalsa, in mitigating radiation-induced side effects, emphasize the underlying mechanism of action for the beneficial effects and address aspects that merit detail investigations for these fruits to move towards clinical application in the near future. METHODS: The authors data-mined Google Scholar, PubMed, Embase, and the Cochrane Library for publications in the field from 1981 up to July 2020. The focus was on the radioprotection and the mechanism responsible for the beneficial effects, and accordingly, the articles were collated and analyzed. RESULTS: This article emphasizes the usefulness of stone apple, mango, Indian gooseberry, black plum, and phalsa as radioprotective agents. From a mechanistic view, reports are suggestive that the beneficial effects are mediated by triggering free radical scavenging, antioxidant, anti-mutagenic and anti-inflammatory effects. CONCLUSION: For the first time, this review addresses the beneficial effects of mango, Indian gooseberry, black plum, stone apple and phalsa as radioprotective agents. The authors suggest that future studies should be directed at understanding the selective radioprotective effects with tumor-bearing laboratory animals to understand their usefulness as radioprotective drug/s during radiotherapy and as a food supplement to protect people from getting exposed to low doses of radiation in occupational settings. Phase I clinical trial studies are also required to ascertain the optimal dose and the schedule to be followed with the standardized extract of these fruits. The most important aspect is that these fruits, being a part of the diet, have been consumed since the beginning of mankind, are non-toxic, possess diverse medicinal properties, have easy acceptability, all of which will help take research forward and be of benefit to patients, occupational workers, agro-based sectors and pharma industries.

Cell-Free Protein Systems from Yersinia pestis are Functional and Growth-Temperature Dependent.

Cellular lysates capable of transcription and translation have become valuable tools for prototyping genetic circuits, screening engineered functional parts, and producing biological components. Here we report that lysates derived from Yersinia pestis CO92- are functional and can utilize both the E. coli σ70 and the bacteriophage T7 promoter systems to produce green fluorescent protein (GFP). Because of the natural lifestyle of Y. pestis, lysates were produced from cultures grown at 21 °C, 26 °C, and 37 °C to mimic the infection cycle. Regardless of the promoter system the GFP production from 37 °C was the most productive and the 26 °C lysate was the least. When reactions are initiated with 5 nM of DNA, the GFP output of the 37 °C lysate is comparable with the productivity of other non-E. coli systems. The data we present demonstrate that, without genetic modification to enhance productivity, cell-free extracts from Y. pestis are functional and dependent on the temperature at which the bacterium was grown.

Blind Spot: A Braille Patterned Novel Multiplex Lateral Flow Immunoassay Sensor Array for the Detection of Biothreat Agents.

Lateral flow immunoassays (LFIs) are simple, point-of-care diagnostic devices used for detecting biological agents or other analytes of interest in a sample. LFIs are predominantly singleplex assays, interrogating one target analyte at a time. There is a need for multiplex LFI devices, e.g., a syndromic panel to differentiate pathogens causing diseases exhibiting similar symptoms. Multiplex LFI devices would be especially valuable in instances where sample quantity is limiting and reducing assay time and costs is critical. There are limitations to the design parameters and performance characteristics of a multiplex LFI assay with many horizontal test lines due to constraints in capillary flow dynamics. To address some of the performance issues, we have developed a spot array multiplex LFI using Braille format (hence called Blind Spot) and a sensor, MACAW (Modular Automated Colorimetric Analyses Widget), that can analyze and interpret the results. As a proof of concept, we created a multiplex toxin panel, for detecting three toxins, using two letter codes for each. The results indicated that the six-plex, triple toxin assay performs as well as singleplex assays. The sensor-based calls are better compared to human interpretation in discriminating and interpreting ambiguous test results correctly especially at lower antigen concentrations and from strips with blemishes.

Generating Biologically Stable TNA Aptamers that Function with High Affinity and Thermal Stability.

Aptamers are often prone to nuclease digestion, which limits their utility in many biomedical applications. Here we describe a xeno-nucleic acid system based on α-l-threofuranosyl nucleic acid (TNA) that is completely refractory to nuclease digestion. The use of an engineered TNA polymerase permitted the isolation of functional TNA aptamers that bind to HIV reverse transcriptase (HIV RT) with KD's of ∼0.4-4.0 nM. The aptamers were identified using a display strategy that provides a powerful genotype-phenotype linkage. The TNA aptamers remain active in the presence of nuclease and exhibit markedly higher thermal stability than monoclonal antibodies. The combined properties of biological stability, high binding affinity, and thermal stability make TNA aptamers a powerful system for the development of diagnostic and therapeutic agents.

The gastric emptying study with oatmeal: reference range and reproducibility as a function of age and sex.

UNLABELLED: This study evaluated the reference range and reproducibility of the gastric emptying study with oatmeal as a function of age and sex. METHODS: Twenty-four healthy subjects, 12 men and 12 women, categorized into 3 age groups, 20-40, 40-60, and 60-80 y, were studied twice, 1 d apart, with instant oatmeal labeled with (99m)Tc-sulfur colloid. Imaging was performed in the upright position using the left anterior oblique (LAO), right posterior oblique (RPO), anterior, and posterior projections. One-minute digital images acquired every 15 min for 60 min were used to calculate a simple half-time of emptying. RESULTS: A strong correlation was found among half-times of gastric emptying calculated from the anterior projection, LAO projection, anterior-posterior geometric mean, and LAO-RPO geometric mean (P < 0.01). A significant inverse correlation was found between increasing age and decreasing half-time of emptying in men and women (P < 0.05). A reference range of 10-60 min is suggested for 20- to 40-y-old patients, 10-40 min for 40- to 60-y-olds, and 10-30 min for 60- to 80-y-olds. Half-times of emptying tended to be longer for women than for men (not statistically significant). There was a large variation between the first and second studies, with a trend toward decreasing variation with increasing age in both men and women. In repeated studies, a reference range of variation of up to 30 min is suggested for 20- to 40-y-old patients, up to 20 min for 40- to 60-y-olds, and up to 15 min for 60- to 80-y-olds. CONCLUSION: The reference range for half-time of gastric emptying with instant oatmeal decreases with increasing age in both men and women. Test-retest variation is relatively large and tends to decrease with increasing age in both men and women. Data from either the LAO projection or the anterior-posterior geometric mean are acceptable for calculating the half-time of gastric emptying.