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The Analyst Jun 2022Malaria was regarded as the most devastating infectious disease of the 21st century until the COVID-19 pandemic. Asexual blood staged parasites (ABS) play a unique role...
Malaria was regarded as the most devastating infectious disease of the 21st century until the COVID-19 pandemic. Asexual blood staged parasites (ABS) play a unique role in ensuring the parasite's survival and pathogenesis. Hitherto, there have been no spectroscopic reports discriminating the life cycle stages of the ABS parasite under physiological conditions. The identification and quantification of the stages in the erythrocytic life cycle is important in monitoring the progression and recovery from the disease. In this study, we explored visible microspectrophotometry coupled to machine learning to discriminate functional ABS parasites at the single cell level. Principal Component Analysis (PCA) showed an excellent discrimination between the different stages of the ABS parasites. Support Vector Machine Analysis provided a 100% prediction for both schizonts and trophozoites, while a 92% and 98% accuracy was achieved for predicting control and ring staged infected RBCs, respectively. This work shows proof of principle for discriminating the life cycle stages of parasites in functional erythrocytes using visible microscopy and thus eliminating the drying and fixative steps that are associated with other optical-based spectroscopic techniques.
Topics: Animals; COVID-19; Erythrocytes; Humans; Life Cycle Stages; Machine Learning; Malaria; Malaria, Falciparum; Microspectrophotometry; Pandemics; Parasites; Plasmodium falciparum
PubMed: 35611958
DOI: 10.1039/d2an00274d -
Animal Reproduction Science Jul 2021Freshwater unionid mussel diversity is decreasing because of species extirpation or extinction. While little can be done to recover lost species, there is an opportunity...
Freshwater unionid mussel diversity is decreasing because of species extirpation or extinction. While little can be done to recover lost species, there is an opportunity to develop techniques to save other species. This can be facilitated through gene banking and assisted reproduction. Unfortunately, limited information is available on mussel reproduction, especially relating to sperm quality. Objectives, therefore, were to quantify seasonal changes in sperm concentration and morphology for two unionid mussels, Ligumia subrostrata and Lampsilisstraminea, measure intraspecific heterogeneity for sperm morphometry, and develop an efficient method to quantify sperm concentration using a microspectrophotometer. There were no differences in sperm concentration when cells were extracted from the center or at a half centimeter on either side of the visceral mass, during the spawning season. There was a seasonal change in sperm concentration, such that concentration for L. subrostrata ranged from 1.1 × 10 to 19.60 × 10 cells/mL with there being the largest counts between 26 September to 7 November. L. straminea sperm concentration was greatest (20.0 × 10 cells/mL) on 13 September and subsequently decreased. Sperm were uniflagellated and SEM results for L. subrostrata and L. straminea showed mean head length and width (mid-spawning) were 3.38 ± 0.04 μm and 1.61 ± 0.01 μm and 3.37 ± 0.04 μm and 1.61 ± 0.01 μm, respectively. There were close (R ≥ 0.85) quadratic associations between hemocytometer counts and absorbance (300, 600, 700 nm). These results provide baseline information to further investigate sperm quality, fertilizing capacity, and cryopreservation for freshwater mussels.
Topics: Animals; Male; Microspectrophotometry; Seasons; Species Specificity; Spermatozoa; Unionidae
PubMed: 34051647
DOI: 10.1016/j.anireprosci.2021.106768 -
Annals of Biomedical Engineering Dec 2019Melanoma is the deadliest type of skin cancer with its prevalence on the rise. Recently, the melanocyte stem cells in hair follicles have been identified as the possible...
Melanoma is the deadliest type of skin cancer with its prevalence on the rise. Recently, the melanocyte stem cells in hair follicles have been identified as the possible origin of melanoma upon exposure to ultraviolet radiation (UVR) through skin. It is hypothesized that colourless vellus hair (predominant in childhood) can serve as an alternative pathway in transmitting these ultraviolet (UV) photons to the stem cells. To investigate this, we have used the CRAIC microspectrophotometer to investigate the optical properties of 'vellus-like' hairs and terminal hairs of different colours using UV-VIS-NIR light sources. It was found that the average attenuation coefficient of 'vellus-like' hair is significantly lower than that of terminal hair in the UVA (p < 0.0001) and UVB (p < 0.001) wavelength ranges. Next, the optical properties of hairs are applied to simulations for examining their influence on UV transmission into the skin. The results show that the presence of vellus hair would increase the solar UV transmission to the melanocyte stem cell layer significantly. The findings explain why children are particularly vulnerable to sun exposure and the positive correlation found between the incidence of melanoma in adults' bodies and the number of vellus hairs in these areas.
Topics: Adult; Child; Child, Preschool; Female; Hair; Humans; Infant; Male; Melanoma; Middle Aged; Skin; Skin Neoplasms; Spectrophotometry; Ultraviolet Rays; Young Adult
PubMed: 31290037
DOI: 10.1007/s10439-019-02315-z -
PloS One 2019Small parasitoid wasps are abundant and extremely diverse, yet their colors have not been analyzed. One of the more common color patterns observed in these wasps is a...
Small parasitoid wasps are abundant and extremely diverse, yet their colors have not been analyzed. One of the more common color patterns observed in these wasps is a black-orange-black pattern, which is especially common among neotropical species of Scelionidae ranging in size from 2 to 10 mm. Due to the methodological challenges involved in extracting and analyzing pigments from small-sized insects, other methods for examining colors need to be explored. In this work, we propose the use of microspectrophotometry in combination with statistical analysis methods in order to 8 study the spectral properties in such cases. We examined 8 scelionid genera and 1 genus from a distantly related family (Evaniidae), all showing the black-orange-black pattern. Functional Data Analysis and statistical analysis of Euclidean distances for color components were applied to study color differences both between and within genera. The Functional Data Analysis proved to be a better method for treating the reflectance data because it gave a better representation of the physical information. Also, the reflectance spectra were separated into spectral color component contributions and each component was labeled according to its own dominant wavelength at the maximum of the spectrum: Red, Green and Blue. When comparing spectral components curves, the spectral blue components of the orange and black colors, independent of the genera being compared, result almost identical, suggesting that there is a common compound for the pigments. The results also suggest that cuticle from different genera, but with the same color might have a similar chemical composition. This is the first time that the black and orange colors in small parasitoid wasps has been analyzed and our results provide a basis for future research on the color patterns of an abundant but neglected group of insects.
Topics: Algorithms; Animals; Color; Models, Biological; Pigmentation; Pigments, Biological; Spectrum Analysis; Wasps
PubMed: 31647807
DOI: 10.1371/journal.pone.0218061 -
Marine Drugs Apr 2021Sulfated galactans (SG) isolated from red alga have been reported to inhibit the growth of cholangiocarcinoma (CCA) cells, which was similar to the epidermal growth... (Comparative Study)
Comparative Study
Sulfated galactans (SG) isolated from red alga have been reported to inhibit the growth of cholangiocarcinoma (CCA) cells, which was similar to the epidermal growth factor receptor (EGFR)-targeted drug, cetuximab. Herein, we studied the anti-cancer potency of SG compared to cetuximab. Biological studies demonstrated SG and cetuximab had similar inhibition mechanisms in CCA cells by down-regulating EGFR/ERK pathway, and the combined treatment induced a greater inhibition effect. The molecular docking study revealed that SG binds to the dimerization domain of EGFR, and this was confirmed by dimerization assay, which showed that SG inhibited ligand-induced EGFR dimer formation. Synchrotron FTIR microspectroscopy was employed to examine alterations in cellular macromolecules after drug treatment. The SR-FTIR-MS elicited similar spectral signatures of SG and cetuximab, pointing towards the bands of RNA/DNA, lipids, and amide I vibrations, which were inconsistent with the changes of signaling proteins in CCA cells after drug treatment. Thus, this study demonstrates the underlined anti-cancer mechanism of SG by interfering with EGFR dimerization. In addition, we reveal that FTIR signature spectra offer a useful tool for screening anti-cancer drugs' effect.
Topics: Antineoplastic Agents; Bile Duct Neoplasms; Cell Line, Tumor; Cell Movement; Cetuximab; Cholangiocarcinoma; ErbB Receptors; Galactans; Humans; Microspectrophotometry; Molecular Docking Simulation; Protein Binding; Protein Multimerization; Signal Transduction; Spectroscopy, Fourier Transform Infrared; Sulfur Compounds; Synchrotrons
PubMed: 33946151
DOI: 10.3390/md19050258 -
The Analyst Mar 2020The use of nanoparticles (NP) as dose enhancers in radiotherapy (RT) is a growing research field. Recently, the use of NP has been extended to charged particle therapy...
Study of the intracellular nanoparticle-based radiosensitization mechanisms in F98 glioma cells treated with charged particle therapy through synchrotron-based infrared microspectroscopy.
The use of nanoparticles (NP) as dose enhancers in radiotherapy (RT) is a growing research field. Recently, the use of NP has been extended to charged particle therapy in order to improve the performance in radioresistant tumors. However, the biological mechanisms underlying the synergistic effects involved in NP-RT approaches are not clearly understood. Here, we used the capabilities of synchrotron-based Fourier Transform Infrared Microspectroscopy (SR-FTIRM) as a bio-analytical tool to elucidate the NP-induced cellular damage at the molecular level and at a single-cell scale. F98 glioma cells doped with AuNP and GdNP were irradiated using several types of medical ion beams (proton, helium, carbon and oxygen). Differences in cell composition were analyzed in the nucleic acids, protein and lipid spectral regions using multivariate methods (Principal Component Analysis, PCA). Several NP-induced cellular modifications were detected, such as conformational changes in secondary protein structures, intensity variations in the lipid CHx stretching bands, as well as complex DNA rearrangements following charged particle therapy irradiations. These spectral features seem to be correlated with the already shown enhancement both in the DNA damage response and in the reactive oxygen species (ROS) production by the NP, which causes cell damage in the form of protein, lipid, and/or DNA oxidations. Vibrational features were NP-dependent due to the NP heterogeneous radiosensitization capability. Our results provided new insights into the molecular changes in response to NP-based RT treatments using ion beams, and highlighted the relevance of SR-FTIRM as a useful and precise technique for assessing cell response to innovative radiotherapy approaches.
Topics: Animals; Cell Line, Tumor; Gadolinium; Light; Lipids; Metal Nanoparticles; Microspectrophotometry; Nucleic Acid Conformation; Nucleic Acids; Principal Component Analysis; Protein Conformation; Proteins; Radiation-Sensitizing Agents; Rats; Silver; Synchrotrons
PubMed: 31993615
DOI: 10.1039/c9an02350j -
Planta Sep 2020EgPHI-1 is a member of PHI-1/EXO/EXL protein family. Its overexpression in tobacco resulted in changes in biomass partitioning, xylem fiber length, secondary cell wall...
EgPHI-1 is a member of PHI-1/EXO/EXL protein family. Its overexpression in tobacco resulted in changes in biomass partitioning, xylem fiber length, secondary cell wall thickening and composition, and lignification. Here, we report the functional characterization of a PHOSPHATE-INDUCED PROTEIN 1 homologue showing differential expression in xylem cells from Eucalyptus species of contrasting phenotypes for wood quality and growth traits. Our results indicated that this gene is a member of the PHI-1/EXO/EXL family. Analysis of the promoter cis-acting regulatory elements and expression responses to different treatments revealed that the Eucalyptus globulus PHI-1 (EgPHI-1) is transcriptionally regulated by auxin, cytokinin, wounding and drought. EgPHI-1 overexpression in transgenic tobacco changed the partitioning of biomass, favoring its allocation to shoots in detriment of roots. The stem of the transgenic plants showed longer xylem fibers and reduced cellulose content, while the leaf xylem had enhanced secondary cell wall thickness. UV microspectrophotometry of individual cell wall layers of fibers and vessels has shown that the transgenic plants exhibit differences in the lignification of S2 layer in both cell types. Taken together, the results suggest that EgPHI-1 mediates the elongation of secondary xylem fibers, secondary cell wall thickening and composition, and lignification, making it an attractive target for biotechnological applications in forestry and biofuel crops.
Topics: Cell Wall; Cellulose; Eucalyptus; Gene Expression Regulation, Plant; Indoleacetic Acids; Lignin; Phylogeny; Plant Proteins; Plant Shoots; Plants, Genetically Modified; Promoter Regions, Genetic; Nicotiana; Xylem
PubMed: 32880001
DOI: 10.1007/s00425-020-03450-x -
Forensic Science International Aug 2021This paper describes the involvement of our laboratory in a Western Australian 'cold-case' investigation that spanned 24 years. The investigation was widely considered...
This paper describes the involvement of our laboratory in a Western Australian 'cold-case' investigation that spanned 24 years. The investigation was widely considered to be the largest in Australian history. During the investigative phase, our laboratory was tasked with the collection of trace evidence of all types in connection with a sexual assault and two homicides that were suspected to be related. Textile fibres represented the vast majority of trace evidence recovered. A much greater quantity of fibres (>10,800) was collected than would be typical for a routine case, as fibres of any colour or type were potentially of investigative value. The investigation was unprecedented in its scale, and presented numerous challenges in terms of evidence recovery, analysis, interpretation, reporting, and provision of testimony. A textile fibre microspectrophotometric (MSP) database was developed specifically for the interpretation of data in connection with the investigation. The database currently contains over 25,000 normalised and first derivative spectra of casework, validation and reference textile fibres. A fibre comparison strategy was devised, involving the identification of preliminary fibre groups on the basis of corresponding/similar MSP spectra, and verification of these groups via brightfield and fluorescence comparison microscopy. A potential link to an automotive source was identified for one of the homicide victims during the investigative phase. After identification of a suspect, a total of 98 fibres recovered from victims and from a seized motor vehicle were found to correspond in properties to six different fibre types from known textile sources in connection with the suspect. A highly publicised criminal trial was held, and textile fibre evidence provided a major contribution to the trial findings, in which the accused was found guilty of two homicides.
Topics: Australia; Databases, Factual; Forensic Sciences; Homicide; Humans; Microspectrophotometry; Spectroscopy, Fourier Transform Infrared; Textiles
PubMed: 34271326
DOI: 10.1016/j.forsciint.2021.110894 -
Science & Justice : Journal of the... Jan 2024In 2017 Cooktown resident Donna Steele was murdered and the case remained unsolved for more than a year. The forensic evidence from the investigation included two...
In 2017 Cooktown resident Donna Steele was murdered and the case remained unsolved for more than a year. The forensic evidence from the investigation included two lengths of synthetic twine and a new protocol for the comparison of twine was developed to enhance the potential value of any evidence. The method was developed using 23 samples of similar twine collected across Australia. Traditional methods of physical and microscopic comparisons and polymer analysis by infrared spectroscopy were retained. Micro-spectrophotometry was used as an objective assessment of colour and was able to identify five groupings within the background samples. Measurements of hydrogen and carbon stable isotopic composition provided further delineation of the background samples. Combining traditional methods with micro-spectrophotometry and stable isotope measurements, the two case samples were found to be distinct from the background population and were indistinguishable when compared to each other.
Topics: Humans; Australia; Carbon Isotopes; Spectrophotometry, Infrared
PubMed: 38182309
DOI: 10.1016/j.scijus.2023.11.006 -
Forensic Science International Sep 2020In their paper "The evaluation of evidence for microspectrophotometry data using functional data analysis", in FSI 305, Aitken et al. present a likelihood-ratio (LR)...
In their paper "The evaluation of evidence for microspectrophotometry data using functional data analysis", in FSI 305, Aitken et al. present a likelihood-ratio (LR) system for their data. We show the values generated by this system cannot be interpreted as LRs: they are ill-calibrated and should be interpreted as discriminating scores. We demonstrate how to transform the scores to well-calibrated LRs using a post-hoc calibrating step. Also, we address criticisms of calibration posited by Aitken et al. We conclude by noting that ill-calibrated LR-values are misleadingly small or large. Therefore calibration should be measured and, if necessary, corrected for. The corrected LR-values (instead of the discriminating scores) can be used to update the prior odds in Bayes rule.
PubMed: 32663721
DOI: 10.1016/j.forsciint.2020.110388