De novo assembly and annotation of the Patagonian toothfish (Dissostichus eleginoides) genome.

BACKGROUND: Patagonian toothfish (Dissostichus eleginoides) is an economically and ecologically important fish species in the family Nototheniidae. Juveniles occupy progressively deeper waters as they mature and grow, and adults have been caught as deep as 2500 m, living on or in just above the southern shelves and slopes around the sub-Antarctic islands of the Southern Ocean. As apex predators, they are a key part of the food web, feeding on a variety of prey, including krill, squid, and other fish. Despite its importance, genomic sequence data, which could be used for more accurate dating of the divergence between Patagonian and Antarctic toothfish, or establish whether it shares adaptations to temperature with fish living in more polar or equatorial climes, has so far been limited. RESULTS: A high-quality D. eleginoides genome was generated using a combination of Illumina, PacBio and Omni-C sequencing technologies. To aid the genome annotation, the transcriptome derived from a variety of toothfish tissues was also generated using both short and long read sequencing methods. The final genome assembly was 797.8 Mb with a N50 scaffold length of 3.5 Mb. Approximately 31.7% of the genome consisted of repetitive elements. A total of 35,543 putative protein-coding regions were identified, of which 50% have been functionally annotated. Transcriptomics analysis showed that approximately 64% of the predicted genes (22,617 genes) were found to be expressed in the tissues sampled. Comparative genomics analysis revealed that the anti-freeze glycoprotein (AFGP) locus of D. eleginoides does not contain any AFGP proteins compared to the same locus in the Antarctic toothfish (Dissostichus mawsoni). This is in agreement with previously published results looking at hybridization signals and confirms that Patagonian toothfish do not possess AFGP coding sequences in their genome. CONCLUSIONS: We have assembled and annotated the Patagonian toothfish genome, which will provide a valuable genetic resource for ecological and evolutionary studies on this and other closely related species.

Surviving winter on the Qinghai-Tibetan Plateau: Pikas suppress energy demands and exploit yak feces to survive winter.

The Qinghai-Tibetan Plateau, with low precipitation, low oxygen partial pressure, and temperatures routinely dropping below -30 °C in winter, presents several physiological challenges to its fauna. Yet it is home to many endemic mammalian species, including the plateau pika (Ochotona curzoniae). How these small animals that are incapable of hibernation survive the winter is an enigma. Measurements of daily energy expenditure (DEE) using the doubly labeled water method show that pikas suppress their DEE during winter. At the same body weight, pikas in winter expend 29.7% less than in summer, despite ambient temperatures being approximately 25 °C lower. Combined with resting metabolic rates (RMRs), this gives them an exceptionally low metabolic scope in winter (DEE/RMRt = 1.60 ± 0.30; RMRt is resting metabolic rate at thermoneutrality). Using implanted body temperature loggers and filming in the wild, we show that this is achieved by reducing body temperature and physical activity. Thyroid hormone (T3 and T4) measurements indicate this metabolic suppression is probably mediated via the thyroid axis. Winter activity was lower at sites where domestic yak (Bos grunniens) densities were higher. Pikas supplement their food intake at these sites by eating yak feces, demonstrated by direct observation, identification of yak DNA in pika stomach contents, and greater convergence in the yak/pika microbiotas in winter. This interspecific coprophagy allows pikas to thrive where yak are abundant and partially explains why pika densities are higher where domestic yak, their supposed direct competitors for food, are more abundant.

Experimental Evaluation of Pedestrian-Induced Multiaxial Gait Loads on Footbridges: Effects of the Structure-to-Human Interaction by Lateral Vibrating Platforms.

The introduction of resistant and lightweight materials in the construction industry has led to civil structures being vulnerable to excessive vibrations, particularly in footbridges exposed to human-induced gait loads. This interaction, known as Human-Structure Interaction (HSI), involves a complex interplay between structural vibrations and gait loads. Despite extensive research on HSI, the simultaneous effects of lateral structural vibrations with fundamental frequencies close to human gait frequency (around 1.0 Hz) and wide amplitudes (over 30.0 mm) remain inadequately understood, posing a contemporary structural challenge highlighted by incidents in iconic bridges like the Millennium Bridge in London, Solferino Bridge in Paris, and Premier Bridge in Cali, Colombia. This paper focuses on the experimental exploration of Structure-to-Human Interaction (S2HI) effects using the Human-Structure Interaction Multi-Axial Test Framework (HSI-MTF). The framework enables the simultaneous measurement of vertical and lateral loads induced by human gait on surfaces with diverse frequency ranges and wide-amplitude lateral harmonic motions. The study involved seven test subjects, evaluating gait loads on rigid and harmonic lateral surfaces with displacements ranging from 5.0 to 50.0 mm and frequency content from 0.70 to 1.30 Hz. A low-cost vision-based motion capture system with smartphones analyzed the support (Tsu) and swing (Tsw) periods of human gait. Results indicated substantial differences in Tsu and Tsw on lateral harmonic protocols, reaching up to 96.53% and 58.15%, respectively, compared to rigid surfaces. Normalized lateral loads (LL) relative to the subject's weight (W0) exhibited a linear growth proportional to lateral excitation frequency, with increased proportionality constants linked to higher vibration amplitudes. Linear regressions yielded an average R2 of 0.815. Regarding normalized vertical load (LV) with respect to W0, a consistent behavior was observed for amplitudes up to 30.0 mm, beyond which a linear increase, directly proportional to frequency, resulted in a 28.3% increment compared to rigid surfaces. Correlation analyses using Pearson linear coefficients determined relationships between structural surface vibration and pedestrian lateral motion, providing valuable insights into Structure-to-Human Interaction dynamics.

Oral cavity and oropharyngeal carcinoma disparities in age and survival in Indigenous and non-Indigenous populations of Queensland.

OBJECTIVES: To investigate the risk and prognosis of oral squamous cell carcinoma (SCC) between Indigenous and non-Indigenous populations of Queensland. MATERIALS AND METHODS: Retrospective analysis of data from the Queensland Cancer Registry (QCR) between the years 1982-2018. Main outcome measures were age at diagnosis and cumulative survival to compare the risk and prognosis of oral SCC between the populations. RESULTS: 9424 patients with self-declared ethnicity were identified with oral SCC from the QCR, with a male to female ratio of 2.56:1. Of these patients, 9132 were non-Indigenous (96.9%) and 292 Indigenous (3.1%). Indigenous people were significantly younger at diagnosis (mean (SD) age 54.3 (10.1) years), compared to 62.0 (12.1) years in non-Indigenous people. Mean survival in the full cohort was 4.3 years (SD: 5.6), with Indigenous people presenting a significant shorter mean survival of 2.0 years (SD: 3.5) when compared with 4.4 years (SD: 5.7) in non-Indigenous people (p < 0.001). CONCLUSIONS: Indigenous Australians are diagnosed at a significantly younger age and present with worse survival and poorer prognosis. Due to missing variables in the Queensland Cancer Registry, it is not possible in the current study to ascertain the scientific or social reasons behind these disparities. CLINICAL RELEVANCE: Results from this study can inform public policy and raise awareness in Queensland regarding disparity in oral cancer prognosis.

Oral cancer in Australia: Rising incidence and worsening mortality.

BACKGROUND: Oral cancer, predominantly squamous cell carcinoma (SCC), is a lethal and deforming disease of rising incidence. Although largely preventable by eliminating harmful tobacco and alcohol risk factor behaviour, 5-year survival rates remain around 50%, primarily due to late presentation of advanced stage disease. Whilst low socio-economic status, regional and remote location and indigenous status are associated with head and neck cancer in general, detailed incidence and demographic data for oral SCC in Australia are limited. This study aimed to characterise the Queensland population at risk of oral SCC development. METHODS: Following ethical approval, the Queensland Cancer Register (QCR) dataset was analysed to determine patterns of incidence, anonymised patient demographics, clinical presentation and outcome data for oral SCC cases diagnosed between 1982 and 2018. RESULTS: Data from 9887 patients were obtained. Mean age at diagnosis was 64.55 years, with a male-to-female ratio of 2.51:1; males were diagnosed at a younger age (p < 0.001). At study census date, 59% of patients had died, with females demonstrating longer mean survival (p < 0.001). Clinicopathological data confirmed that SCC most commonly arose from tongue sites (49%) and, whilst tumours were predominantly moderately differentiated in nature (63%), patients with poorly differentiated carcinomas exhibited shortest survival times (p < 0.05). Over the 36-year study period, the number of diagnoses increased 4.49-fold, whilst the number of deaths increased 19.14-fold. CONCLUSION: Oral SCC poses a significant and growing healthcare problem in Queensland. In the absence of national screening, characterising the high-risk oral SCC population facilitates pragmatic opportunities to raise disease awareness, to deliver targeted screening and effective primary prevention strategies, and to provide early interventional treatment intervention to reduce disease mortality and morbidity.

Saliva-based cell-free DNA and cell-free mitochondrial DNA in head and neck cancers have promising screening and early detection role.

BACKGROUND: Cell-free DNA (cfDNA) and cell-free mitochondrial DNA (cf-mtDNA) have been postulated as potential diagnostic and prognostic biomarkers for different human malignancies. Early detection of head and neck malignancies is fundamental for optimal patient management. This study, therefore, aimed to assess the utility of saliva-based liquid biopsy as a noninvasive source of cfDNA and cf-mtDNA for detecting head and neck cancer (HNSCC). METHODS: One hundred thirty-three patients diagnosed with either oral leukoplakia (OLK) or HNSCC were compared with 137 healthy volunteers. An unstimulated whole saliva sample was collected from each participant. The absolute copy numbers of salivary cf-mtDNA and cfDNA were quantified using Multiplex Quantitative PCR. Two diagnostic indices based on the investigated molecules were assessed for their ability to differentiate between different diagnostic categories. RESULTS: The median scores of cfDNA and cf-mtDNA were statistically significantly higher among HNSCC patients (p < 0.05), revealing area under the curve values of 0.758 and 0.826, respectively. The associated accuracy for this test in discriminating HNSCC from other diagnostic categories was 77.37% for the cfDNA-based index and 80.5% for the cf-mtDNA-based index. The median score of cfDNA was statistically significantly higher for patients with severe epithelial dysplasia (OED) compared to those with epithelial keratosis with no OED and mild OED. However, there was no significant difference between controls and OLK individuals. CONCLUSION: cfDNA and cf-mtDNA showed potential for use as precision medicine tools to detect HNSCC. Further multi-centre prospective studies are warranted to assess the prognostic utility of these molecules.

Predicting oral cancer survival-Development and validation of an Asia-Pacific nomogram.

BACKGROUND: Nomograms are graphical calculating devices that predict response to treatment during cancer management. Oral squamous cell carcinoma (OSCC) is a lethal and deforming disease of rising incidence and global significance. The aim of this study was to develop a nomogram to predict individualized OSCC survival using a population-based dataset obtained from Queensland, Australia and externally validated using a cohort of OSCC patients treated in Hong Kong. METHODS: Clinico-pathological data for newly diagnosed OSCC patients, including age, sex, tumour site and grading, were accessed retrospectively from the Queensland Cancer Registry (QCR) in Australia and the Clinical Data Analysis and Reporting System (CDARS) in Hong Kong. Multivariate Cox proportional hazard regression was used to construct overall survival (OS) and cancer-specific survival (CSS) prediction models. Nomograms were internally validated using 10-fold cross validation, and externally validated against the Hong Kong dataset. RESULTS: Data from 9885 OSCC patients in Queensland and 465 patients from Hong Kong were analysed. All clinico-pathological variables significantly influenced survival outcomes. Nomogram calibration curves demonstrated excellent agreement between predicted and actual probability for Queensland patients. External validation in the Hong Kong population demonstrated slightly poorer nomogram performance, but predictive power remained strong. CONCLUSION: Based upon readily available data documenting patient demographic and clinico-pathological variables, predictive nomograms offer pragmatic aid to clinicians in individualized treatment planning and prognosis assessment in contemporary OSCC management.

Clinical utility of nailfold capillaroscopy.

Nailfold capillaroscopy (NFC) is a simple noninvasive microscopic technique used to identify characteristic morphological abnormalities in the nailfold capillaries. The presence of this microvasculopathy appears to be of fundamental importance in the pathological processes that underlie the scleroderma spectrum disorders (including dermatomyositis and antisynthetase myositis). This review discusses the different methodologies and techniques in performing NFC and stresses the diagnostic utility achieved with simple 'bedside' techniques utilising the ophthalmoscope, dermatoscope or smart phone. Recent advances in reporting abnormal microvascular patterns and vascular metrics (e.g. capillary density and dropout) are discussed. The aetiopathogenesis of the microvasculopathy is currently unknown but its close association with Raynaud Phenomena and specific autoantibodies together with recent observations from sequential NFC allows speculations on its possible mechanism. Finally, future developments in the use of NFC as a possible biomarker in the management of the scleroderma spectrum disorders are discussed, with a recommendation that NFC becomes more widely available, particularly in rheumatological, immunological and dermatological practice. NFC provides a clinically accessible window on the pathologic process fundamental to scleroderma-related disease.

The Impact of Vaccination on Incidence and Outcomes of SARS-CoV-2 Infection in Patients with Kidney Failure in Scotland.

BACKGROUND: Patients with kidney failure requiring KRT are at high risk of complications and death following SARS-CoV-2 infection, with variable antibody responses to vaccination reported. We investigated the effects of COVID-19 vaccination on the incidence of infection, hospitalization, and death from COVID-19 infection. METHODS: The study design was an observational data linkage cohort study. Multiple health care datasets were linked to ascertain all SARS-CoV-2 testing, vaccination, hospitalization, and mortality data for all patients treated with KRT in Scotland from the start of the pandemic over a period of 20 months. Descriptive statistics, survival analyses, and vaccine effectiveness were calculated. RESULTS: As of September 19, 2021, 93% (n=5281) of the established KRT population in Scotland had received two doses of an approved SARS-CoV-2 vaccine. Over the study period, there were 814 cases of SARS-CoV-2 infection (15.1% of the KRT population). Vaccine effectiveness rates against infection and hospitalization were 33% (95% CI, 0 to 52) and 38% (95% CI, 0 to 57), respectively. Within 28 days of a SARS-CoV-2-positive PCR test, 9.2% of fully vaccinated individuals died (7% patients on dialysis and 10% kidney transplant recipients). This compares to <0.1% of the vaccinated general Scottish population admitted to the hospital or dying due to COVID-19 during that period. CONCLUSIONS: These data demonstrate that a primary vaccine course of two doses has limited effect on COVID-19 infection and its complications in patients with KRT. Adjunctive strategies to reduce risk of both COVID-19 infection and its complications in this population are urgently required.

Estimating heritability using family-pooled phenotypic and genotypic data: a simulation study applied to aquaculture.

Estimating heritability based on individual phenotypic and genotypic measurements can be expensive and labour-intensive in commercial aquaculture breeding. Here, the feasibility of estimating heritability using within-family means of phenotypes and allelic frequencies was investigated. Different numbers of full-sib families and family sizes across ten generations with phenotypic and genotypic information on 10 K SNPs were analysed in ten replicates. Three scenarios, representing differing numbers of pools per family (one, two and five) were considered. The results showed that using one pool per family did not reliably estimate the heritability of family means. Using simulation parameters appropriate for aquaculture, at least 200 families of 60 progeny per family divided equally in two pools per family was required to estimate the heritability of family means effectively. Although application of five pools generated more within- and between- family relationships, it reduced the number of individuals per pool and increased within-family residual variation, hence, decreased the heritability of family means. Moreover, increasing the size of pools resulted in increasing the heritability of family means towards one. In addition, heritability of family mean estimates were higher than family heritabilities obtained from Falconer's formula due to lower intraclass correlation estimate compared to the coefficient of relationship.

The clinical utility of contemporary oral epithelial dysplasia grading systems.

INTRODUCTION: Clinical management of oral potentially malignant disorders relies on accurate histopathological assessment of the presence and grade of oral epithelial dysplasia. While adjunctive laboratory tests have provided useful prognostic information, none are in widespread clinical use. This study was performed to assess the clinical utility of two contemporary oral epithelial dysplasia grading systems. METHODS: Patients were identified from a clinical database. Oral epithelial dysplasia grading was performed by three oral and maxillofacial pathologists blinded to clinical outcome using the WHO 2017 system and a binary classification. The primary outcome measure was the development of oral squamous cell carcinoma, termed 'malignant transformation'. RESULTS: One hundred thirty-one cases satisfied the inclusion criteria, of which 23 underwent malignant transformation. There was substantial inter-rater agreement between the study pathologists for both grading systems, measured using kappa statistics (κ = 0.753 - 0.784). However, there was only moderate agreement between the consensus WHO 2017 dysplasia grade for the study against the original grade assigned by a pool of six pathologists in the context of the clinical service (κ = 0.491). Higher grade categories correlated with an increased risk of developing cancer using both grading systems. CONCLUSION: This study demonstrates that the WHO 2017 and binary grading systems are reproducible between calibrated pathologists and that consensus reporting is likely to improve the consistency of grading. The WHO and binary systems were prognostically comparable. We recommend that institutions implement consensus oral epithelial dysplasia grading and prospectively audit the effectiveness of risk stratifying their patients with oral potentially malignant disorders. (249 words).

A comparison of two methods for the detection of circulating tumour cells in patients with oral cavity cancer.

BACKGROUND: Circulating tumour cells (CTCs) detected in patient blood samples are relevant as diagnostic and prognostic markers offering insights into tumour behaviour and guiding treatment of cancer at an individualised level. The aim of this study was to ascertain the feasibility of detecting CTCs in oral squamous cell carcinoma (OSCC) using two different methods so as to determine the optimal method for the study of this cancer. METHODS: Comparison of the numbers of CTCs, circulating tumour micro-emboli (CTMs) and circulating tumour endothelial cells (CTECs), was undertaken in forty clinical samples of oral squamous cell carcinoma (OSCC) determined by filtration (ISET® ) and in situ fluorescent immunostaining (i-FISH, Cytelligen® ) immunostaining and in situ hybridisation. RESULTS: i-FISH detected CTCs in 80% of samples compared with 40% of samples analysed by microfiltration. i-FISH detected CTCs in a further 40% of samples in which microfiltration did not detect CTCs. No CTC clusters were detected by microfiltration while i-FISH detected CTM in 12.5% of samples. i-FISH analysis detected CTECs in 20/40 samples. CONCLUSION: These results highlight significant differences in detection of CTCs, CTM and CTECs between i-FISH and microfiltration when applied to OSCC samples, suggesting that technologies capable of detecting circulating aneuploid cells more accurately detect CTCs. i-FISH also detected CTM and CTEC not detected using ISET® . With proven prognostic relevance in adenocarcinomas, accurate enumeration of CTCs, CTMs and CTECs may be a clinically useful tool in the management of OSCC and may aid in the reduction of false-negative diagnoses.

Performance of a simplified scoring system for risk stratification in oral cancer and oral potentially malignant disorders screening.

BACKGROUND: Impact and efficiency of oral cancer and oral potentially malignant disorders screening are most realized in "at-risk" individuals. However, tools that can provide essential knowledge on individuals' risks are not applied in risk-based screening. This study aims to optimize a simplified risk scoring system for risk stratification in organized oral cancer and oral potentially malignant disorders screening. METHODS: Participants were invited to attend a community-based oral cancer and oral potentially malignant disorders screening program in Hong Kong. Visual oral examination was performed for all attendees and information on sociodemographic characteristics as well as habitual, lifestyle, familial, and comorbidity risk factors were obtained. Individuals' status of those found to have suspicious lesions following biopsy and histopathology were classified as positive/negative and this outcome was used in a multiple logistic regression analysis with variables collected during screening. Odds ratio weightings were then used to develop a simplified risk scoring system which was validated in an external cohort. RESULTS: Of 979 participants, 4.5% had positive status following confirmatory diagnosis. A 12-variable simplified risk scoring system with weightings was generated with an AUC, sensitivity, and specificity of 0.82, 0.71, and 0.78 for delineating high-risk cases. Further optimization on the validation cohort of 491 participants yielded a sensitivity and specificity of 0.75 and 0.87 respectively. CONCLUSIONS: The simplified risk scoring system was able to stratify oral cancer and oral potentially malignant disorders risk with satisfactory sensitivity and specificity and can be applied in risk-based disease screening.

Efficacy of hypermethylated DNA biomarkers in saliva and oral swabs for oral cancer diagnosis: Systematic review and meta-analysis.

OBJECTIVES: This study aims to determine the diagnostic test accuracy (DTA) of hypermethylated DNA biomarkers in saliva and oral swabs for oral squamous cell carcinoma (OSCC) detection from the prevalidation studies available. MATERIALS AND METHODS: Electronic database searching of PubMed, EMBASE, Cochrane Library, Scopus, Web of Science, and LILACS was conducted to identify relevant articles that were published between January 1, 2000, and August 1, 2020. RESULTS: Meta-analysis was conducted based on 11 of 20 studies selected for review. Included studies had high bias concerns on the QUADAS-2 study assessment tool. We found that salivary and oral swab hypermethylation markers had better specificity than sensitivity for oral cancer detection. Summary sensitivity and specificity (95% CI) of hypermethylation panels were 86.2% (60-96.2) and 90.6% (85.9-93.9) while for individual markers, summary sensitivity and specificity (95% CI) were 70% (56.9-80.5) and 91.9% (80.3-96.9), respectively. Respective positive and negative likelihood ratios for combined markers were 9.2 (5.89-14.36) and 0.15 (0.05-0.5), and 8.61 (3.39-21.87) and 0.33 (0.22-0.49) for single-application biomarkers. CONCLUSION: DNA hypermethylation biomarkers especially in combination have acceptable DTA that warrants further optimization with rigorous biomarker evaluation methods for conclusive determination of their efficacy.

Oral Cancer Awareness and Individuals' Inclination to Its Screening and Risk Prediction in Hong Kong.

Assessing the baseline knowledge status and expectations of the target population of any health promotion and secondary prevention program is essential to the success of such intervention. To obtain this information about the Hong Kong population a priori to implementing these preventive strategies for oral cancer in addition to determining the willingness of potential screening participants to take risk-profiling assessments, a cross-sectional survey was conducted between November 2019 and March 2020. A total of 964 residents between the ages 18 and 86 years were invited to participate in this study across the three geographical areas in Hong Kong. Most participants self-reported being aware of oral cancer (86.3%), although the proportion of those with substantial knowledge on salient risk factors and early identifiable signs were very low (2.9%). Age and level of education were the only demographic characteristics associated with the knowledge status. The proportion of participants willing to attend community screening and partake in risk profiling assessment was high (83.9% and 80.9% respectively). Willingness to attend community screening was directly associated with respondents' self-reported oral cancer awareness status (OR: 1.9, 95% CI: 1.22-2.96). Also, we observed that those participants who were willing to attend screening are more inclined to take risk prediction assessments that those not willing to attend. These findings have showcased the need to intensify health promotion via personal skills development to encourage early disease presentation and will assist in the planning of these programs accordingly in the Hong Kong population.

Predicting the clinical outcome of oral potentially malignant disorders using transcriptomic-based molecular pathology.

BACKGROUND: This study was undertaken to develop and validate a gene expression signature that characterises oral potentially malignant disorders (OPMD) with a high risk of undergoing malignant transformation. METHODS: Patients with oral epithelial dysplasia at one hospital were selected as the 'training set' (n = 56) whilst those at another hospital were selected for the 'test set' (n = 66). RNA was extracted from formalin-fixed paraffin-embedded (FFPE) diagnostic biopsies and analysed using the NanoString nCounter platform. A targeted panel of 42 genes selected on their association with oral carcinogenesis was used to develop a prognostic gene signature. Following data normalisation, uni- and multivariable analysis, as well as prognostic modelling, were employed to develop and validate the gene signature. RESULTS: A prognostic classifier composed of 11 genes was developed using the training set. The multivariable prognostic model was used to predict patient risk scores in the test set. The prognostic gene signature was an independent predictor of malignant transformation when assessed in the test set, with the high-risk group showing worse prognosis [Hazard ratio = 12.65, p = 0.0003]. CONCLUSIONS: This study demonstrates proof of principle that RNA extracted from FFPE diagnostic biopsies of OPMD, when analysed on the NanoString nCounter platform, can be used to generate a molecular classifier that stratifies the risk of malignant transformation with promising clinical utility.

Mesophyll conductance in two cultivars of wheat grown in glacial to super-elevated CO2 concentrations.

Mesophyll conductance (gm) is an important factor limiting photosynthesis. However, gm response to long-term growth in variable [CO2] is not well understood, particularly in crop plants. Here, we grew two cultivars of wheat (Halberd and Cranbrook), known to differ in gm under current environmental conditions, in four [CO2] treatments: glacial (206 µmol mol-1), pre-industrial (344 µmol mol-1), current ambient (489 µmol mol-1), and super-elevated (1085 µmol mol-1), and two water treatments (well-watered and moderate water limitation), to develop an evolutionary and future climate perspective on gm control of photosynthesis and water-use efficiency (WUE). In the two wheat genotypes, gm increased with rising [CO2] from glacial to ambient [CO2], but declined at super-elevated [CO2]. The responses of gm to different growth [CO2] also depend on water stress; however, the specific mechanism of gm response to [CO2] remains unclear. Although gm and gm/gsc (mesophyll conductance/stomatal conductance) were strongly associated with the variability of photosynthetic rates (A) and WUE, we found that plants with higher gm may increase A without increasing gsc, which increased WUE. These results may be useful to inform plant breeding programmes and cultivar selection for Australian wheat under future environmental conditions.

Statistical profiling of oral cancer and the prediction of outcome.

BACKGROUND: The global burden of oral squamous cell carcinoma (OSCC) remains formidable. Identifying factors predictive of aggressive tumour behaviour, disease progression and reduced survival time may assist in early identification of "high-risk" patients and appropriately target combination cancer therapies. METHODS: A retrospective review of 467 OSCC patients treated over a 19-year period facilitated detailed clinico-pathological database analysis and determination of clinical outcome categories based upon time to progressive disease (loco-regional tumour recurrence and/or distant metastasis), overall death and OSCC-related death (death directly attributable to OSCC). Odds ratio (OR) and hazard ratio (HR) statistical measures were used to investigate relationships between patient demographics and clinico-pathological tumour features with clinical outcome. RESULTS: Older age at presentation (P = .002) and a history of previous non-head and neck cancer (P = .010) increased the risk of overall death. OR for progressive disease development (P = .008) and OSCC-related death (P = .019) was most significant for buccal tumours. HR confirmed advanced-stage disease increased the risk of progressive disease (P < .001), overall death (P < .001) and OSCC-related death (P < .001). Positive resection margins were associated with a higher risk of OSCC-related death (P = .023). Significantly lower risks of progressive disease development (P = .002) and OSCC-related death (P = .012) were seen in patients undergoing neck dissection, whilst combination chemoradiotherapy reduced HR for overall death (P < .001) and OSCC-related death (P = .011). CONCLUSION: Statistical profiling of OSCC clinico-pathological data identifies significant influences on clinical outcome. This study adds evidence to the hypothesis that buccal SCC displays aggressive tumour behaviour and poor clinical outcome.

Multiple tumour recurrence in oral, head and neck cancer: Characterising the patient journey.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the 15th most common cause of cancer-related mortality worldwide and approximately one oral cancer-related death occurs for every two new diagnoses. Death-due-to-disease is usually ascribed to inoperable primary tumours, treatment complications, second primary tumours arising due to field cancerization, or locoregional recurrence and distant metastases. METHODS: A retrospective review of OSCC patients treated over a 19-year period, betweenOctober 1st , 2000 and October 1st , 2019. Patient demographic records were collected from consecutively treated adult patients with clinical subtypes corresponding to ICD-10 C00-C06, C09 and C10 were retrieved from the database. Patients who had suffered three or more recurrences after diagnosis of the primary tumour are defined as multiple-recurrent patients. RESULTS: A total of 467 OSCC patients were treated during the study period. One hundred and fifty-five patients developed recurrent OSCC, amongst which 22 were designated as multiple cases. The time between initial OSCC diagnosis and first tumour recurrence varied from 3 to 276 months. Nine of the 22 multiple patients (41%) were diagnosed with buccal mucosal SCC as the primary tumour, which is significantly higher than the average prevalence (or 4.4, 95% CI (1.8, 10.8), p < 0.001) for buccal tumours within the cohort. All patients were treated initially by surgical tumour excision. There were no demonstrable differences in adjuvant chemo-radiotherapy regimes in any of the study groups. CONCLUSION: Multiple OSCC development may occur either synchronously or metachronously during the course of oral cancer disease and poses an important management problem in contemporary oncology practice.

Dysplasia and DNA ploidy to prognosticate clinical outcome in oral potentially malignant disorders.

BACKGROUND: Oral potentially malignant disorders are a clinical conundrum as there are no reliable methods to predict their behaviour. We combine conventional oral epithelial dysplasia grading with DNA ploidy analysis to examine the validity of this approach to risk assessment in a cohort of patients with known clinical outcomes. METHODS: Sections from diagnostic biopsies were assessed for oral epithelial dysplasia using the WHO grading system, and DNA ploidy analysis was performed using established methods. Patients reviewed for a minimum of 5 years who did not develop oral squamous cell carcinoma were classified as "non-transforming" cases. Patients that developed oral squamous cell carcinoma ≥ 6 months after the initial diagnostic biopsy were classified as having "malignant transformation." RESULTS: Ninety cases were included in the study. Seventy cases yielded informative DNA ploidy results. Of these 70 cases, 31 progressed to cancer. Oral epithelial dysplasia grading and DNA ploidy status were both significantly associated with clinical outcome (P < 0.05). Severe dysplasia had a hazard ratio of 3.50 (CI: 1.46, 8.45; P = 0.005) compared to cases with mild dysplasia. Aneuploidy had a hazard ratio of 2.09 (CI: 1.01, 4.32; P = 0.046) compared to cases with a diploid/tetraploid status. Receiver operating characteristic analysis gave an area under the curve of 0.617 for DNA ploidy status and 0.688 when DNA ploidy status was combined with dysplasia grading. CONCLUSION: Our findings suggest that combining dysplasia grading with DNA ploidy status has clinical utility which could be used to develop novel management algorithms.