Tγδ LGLL identifies a subset with more symptomatic disease: analysis of an international cohort of 137 patients.

Tγδ large granular lymphocyte leukemia (LGLL) is a rare variant of T-cell LGLL (T-LGLL) that has been less investigated as compared with the more frequent Tαß LGLL, particularly in terms of frequency of STAT3 and STAT5b mutations. In this study, we characterized the clinical and biological features of 137 patients affected by Tγδ LGLL; data were retrospectively collected from 1997 to 2020 at 8 referral centers. Neutropenia and anemia were the most relevant clinical features, being present in 54.2% and 49.6% of cases, respectively, including severe neutropenia and anemia in ∼20% of cases each. Among the various treatments, cyclosporine A was shown to provide the best response rates. DNA samples of 97 and 94 cases were available for STAT3 and STAT5b mutation analysis, with 38.1% and 4.2% of cases being mutated, respectively. Clinical and biological features of our series of Tγδ cases were also compared with a recently published Tαß cohort including 129 cases. Though no differences in STAT3 and STAT5b mutational frequency were found, Tγδ cases more frequently presented with neutropenia (P = .0161), anemia (P < .0001), severe anemia (P = .0065), and thrombocytopenia (P = .0187). Moreover, Vδ2- cases displayed higher frequency of symptomatic disease. Overall, Tγδ cases displayed reduced survival with respect to Tαß cases (P = .0017). Although there was no difference in STAT3 mutation frequency, our results showed that Tγδ LGLL represents a subset of T-LGLL characterized by more frequent symptoms and reduced survival as compared with Tαß LGLL.

ACE2 and TMPRSS2 SARS-CoV-2 infectivity genes: deep mutational scanning and characterization of missense variants.

The human angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) proteins play key roles in the cellular internalization of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the coronavirus responsible for the coronavirus disease of 2019 (COVID-19) pandemic. We set out to functionally characterize the ACE2 and TMPRSS2 protein abundance for variant alleles encoding these proteins that contained non-synonymous single-nucleotide polymorphisms (nsSNPs) in their open reading frames (ORFs). Specifically, a high-throughput assay, deep mutational scanning (DMS), was employed to test the functional implications of nsSNPs, which are variants of uncertain significance in these two genes. Specifically, we used a 'landing pad' system designed to quantify the protein expression for 433 nsSNPs that have been observed in the ACE2 and TMPRSS2 ORFs and found that 8 of 127 ACE2, 19 of 157 TMPRSS2 isoform 1 and 13 of 149 TMPRSS2 isoform 2 variant proteins displayed less than ~25% of the wild-type protein expression, whereas 4 ACE2 variants displayed 25% or greater increases in protein expression. As a result, we concluded that nsSNPs in genes encoding ACE2 and TMPRSS2 might potentially influence SARS-CoV-2 infectivity. These results can now be applied to DNA sequence data for patients infected with SARS-CoV-2 to determine the possible impact of patient-based DNA sequence variation on the clinical course of SARS-CoV-2 infection.

Mycobacterium tuberculosis-induced miR-155 subverts autophagy by targeting ATG3 in human dendritic cells.

Autophagy is a primordial eukaryotic pathway, which provides the immune system with multiple mechanisms for the elimination of invading pathogens including Mycobacterium tuberculosis (Mtb). As a consequence, Mtb has evolved different strategies to hijack the autophagy process. Given the crucial role of human primary dendritic cells (DC) in host immunity control, we characterized Mtb-DC interplay by studying the contribution of cellular microRNAs (miRNAs) in the post-transcriptional regulation of autophagy related genes. From the expression profile of de-regulated miRNAs obtained in Mtb-infected human DC, we identified 7 miRNAs whose expression was previously found to be altered in specimens of TB patients. Among them, gene ontology analysis showed that miR-155, miR-155* and miR-146a target mRNAs with a significant enrichment in biological processes linked to autophagy. Interestingly, miR-155 was significantly stimulated by live and virulent Mtb and enriched in polysome-associated RNA fraction, where actively translated mRNAs reside. The putative pair interaction among the E2 conjugating enzyme involved in LC3-lipidation and autophagosome formation-ATG3-and miR-155 arose by target prediction analysis, was confirmed by both luciferase reporter assay and Atg3 immunoblotting analysis of miR-155-transfected DC, which showed also a consistent Atg3 protein and LC3 lipidated form reduction. Late in infection, when miR-155 expression peaked, both the level of Atg3 and the number of LC3 puncta per cell (autophagosomes) decreased dramatically. In accordance, miR-155 silencing rescued autophagosome number in Mtb infected DC and enhanced autolysosome fusion, thereby supporting a previously unidentified role of the miR-155 as inhibitor of ATG3 expression. Taken together, our findings suggest how Mtb can manipulate cellular miRNA expression to regulate Atg3 for its own survival, and highlight the importance to develop novel therapeutic strategies against tuberculosis that would boost autophagy.

Circulating miRNA-375 as a potential novel biomarker for active Kaposi's sarcoma in AIDS patients.

The aim of this study was to identify circulating microRNAs (miRNAs) that could be used as biomarkers in patients at risk for or affected by AIDS-Kaposi's sarcoma (KS). Screening of 377 miRNAs was performed using low-density arrays in pooled plasma samples of 10 HIV/human herpesvirus 8 (HHV8)-infected asymptomatic and 10 AIDS-KS patients before and after successful combined antiretroviral therapy (cART). MiR-375 was identified as a potential marker of active KS, being the most down-regulated in AIDS-KS patients after cART and the most up-regulated in naïve AIDS-KS patients compared to naïve asymptomatic subjects. Validation on individual plasma samples confirmed that miR-375 levels were higher in AIDS-KS compared to asymptomatic patients, decreased after cART-induced remission in most AIDS-KS patients and increased in patients with active KS. In asymptomatic patients miR-375 was up-regulated after cART in both screening and validation. Statistical analyses revealed an association between miR-375 changes and CD4 cell counts, which could explain the discordant cases and the opposite trend between asymptomatic and AIDS-KS patients. These data suggest that circulating miR-375 might be a good indicator of active AIDS-KS. Moreover, changes in miR-375 levels may have a prognostic value in HIV/HHV8-infected patients undergoing treatment. Further large-scale validation is needed.

Early Intervention to Improve Sucking in Preterm Newborns: A Systematic Review of Quantitative Studies.

BACKGROUND: Premature birth is associated with feeding difficulties due to inadequate coordination of sucking, swallowing, and breathing. Nonnutritive sucking (NNS) and oral stimulation interventions may be effective for oral feeding promotion, but the mechanisms of the intervention effects need further clarifications. PURPOSE: We reviewed preterm infant intervention studies with quantitative outcomes of sucking performance to summarize the evidence of the effect of interventions on specific components of sucking. METHODS: PubMed, CINAHL, MEDLINE, EMBASE, and PSYCOLIST databases were searched for English language publications through August 2017. Studies were selected if they involved preterm infants, tested experimental interventions to improve sucking or oral feeding skills, and included outcome as an objective measure of sucking performance. Specific Medical Subject Headings (MeSH) terms were utilized. RESULTS: Nineteen studies were included in this review: 15 randomized, 1 quasi-randomized, and 3 crossover randomized controlled trials. Intervention types were grouped into 6 categories (i) NNS, (ii) NNS with auditory reinforcement, (iii) sensorimotor stimulation, (iv) oral support, (v) combined training, and (vi) nutritive sucking. Efficiency parameters were positively influenced by most types of interventions, though appear to be less affected by trainings based on NNS alone. IMPLICATIONS FOR PRACTICE: These findings may be useful in the clinical care of infants requiring support to achieve efficient sucking skills through NNS and oral stimulation interventions. IMPLICATIONS FOR RESEARCH: Further studies including quantitative measures of sucking performance outcome measures are needed in order to best understand the needs and provide more tailored interventions to preterm infants.

A coordinate deregulation of microRNAs expressed in mucosa adjacent to tumor predicts relapse after resection in localized colon cancer.

Up to 20% of colorectal cancer (CRC) node-negative patients develop loco-regional or distant recurrences within 5 years from surgery. No predictive biomarker able to identify the node-negative subjects at high risk of relapse after curative treatment is presently available.Forty-eight localized (i.e. stage I-II) colon cancer patients who underwent radical tumor resection were considered. The expression of five miRNAs, involved in CRC progression, was investigated by qRT-PCR in both tumor tissue and matched normal colon mucosa.Interestingly, we found that the coordinate deregulation of four miRNAs (i.e. miR-18a, miR-21, miR-182 and miR-183), evaluated in the normal mucosa adjacent to tumor, is predictive of relapse within 55 months from curative surgery.Our results, if confirmed in independent studies, may help to identify high-risk patients who could benefit most from adjuvant therapy. Moreover, this work highlights the importance of extending the search for tissue biomarkers also to the tumor-adjacent mucosa.

Reconstruction of gene regulatory modules from RNA silencing of IFN-α modulators: experimental set-up and inference method.

BACKGROUND: Inference of gene regulation from expression data may help to unravel regulatory mechanisms involved in complex diseases or in the action of specific drugs. A challenging task for many researchers working in the field of systems biology is to build up an experiment with a limited budget and produce a dataset suitable to reconstruct putative regulatory modules worth of biological validation. RESULTS: Here, we focus on small-scale gene expression screens and we introduce a novel experimental set-up and a customized method of analysis to make inference on regulatory modules starting from genetic perturbation data, e.g. knockdown and overexpression data. To illustrate the utility of our strategy, it was applied to produce and analyze a dataset of quantitative real-time RT-PCR data, in which interferon-α (IFN-α) transcriptional response in endothelial cells is investigated by RNA silencing of two candidate IFN-α modulators, STAT1 and IFIH1. A putative regulatory module was reconstructed by our method, revealing an intriguing feed-forward loop, in which STAT1 regulates IFIH1 and they both negatively regulate IFNAR1. STAT1 regulation on IFNAR1 was object of experimental validation at the protein level. CONCLUSIONS: Detailed description of the experimental set-up and of the analysis procedure is reported, with the intent to be of inspiration for other scientists who want to realize similar experiments to reconstruct gene regulatory modules starting from perturbations of possible regulators. Application of our approach to the study of IFN-α transcriptional response modulators in endothelial cells has led to many interesting novel findings and new biological hypotheses worth of validation.

A circulating miRNA assay as a first-line test for prostate cancer screening.

BACKGROUND: Prostate cancer (PCa) screening currently relies on prostate-specific antigen (PSA) testing and digital rectal examination. However, recent large-scale studies have questioned the long-term efficacy of these tests, and biomarkers that accurately identify PCa are needed. METHODS: We analysed the levels of circulating microRNAs (miRNAs) in patients with elevated PSA who were diagnosed with either localised PCa (n=36) or benign prostatic hyperplasia (BPH, n=31) upon biopsy. Real-time RT-PCR with Taqman probes was used to measure plasma levels of miRNAs. To circumvent problems associated with circulating miRNA quantitation, we computed the expression ratios of upregulated and downregulated miRNAs. RESULTS: The miR-106a/miR-130b and miR-106a/miR-223 ratios were significantly different between the biopsy-positive and BPH groups (P<0.0001), and yielded statistical power values that were >0.99. Both miRNA ratios were highly sensitive and more specific than PSA in discriminating localised PCa from BPH. Receiver operating characteristic curve analysis revealed area under curve values of 0.81 (miR-106a/miR-130b) and 0.77 (miR-106a/miR-223). CONCLUSIONS: Testing for circulating miR-106a/miR-130b and miR-106a/miR-223 ratios may reduce the costs and morbidity of unnecessary biopsies and is feasible for large-scale screening, as it requires measuring only three miRNAs.

Small noncoding RNAs in cells transformed by human T-cell leukemia virus type 1: a role for a tRNA fragment as a primer for reverse transcriptase.

UNLABELLED: The present study employed mass sequencing of small RNA libraries to identify the repertoire of small noncoding RNAs expressed in normal CD4(+) T cells compared to cells transformed with human T-cell leukemia virus type 1 (HTLV-1), the causative agent of adult T-cell leukemia/lymphoma (ATLL). The results revealed distinct patterns of microRNA expression in HTLV-1-infected CD4(+) T-cell lines with respect to their normal counterparts. In addition, a search for virus-encoded microRNAs yielded 2 sequences that originated from the plus strand of the HTLV-1 genome. Several sequences derived from tRNAs were expressed at substantial levels in both uninfected and infected cells. One of the most abundant tRNA fragments (tRF-3019) was derived from the 3' end of tRNA-proline. tRF-3019 exhibited perfect sequence complementarity to the primer binding site of HTLV-1. The results of an in vitro reverse transcriptase assay verified that tRF-3019 was capable of priming HTLV-1 reverse transcriptase. Both tRNA-proline and tRF-3019 were detected in virus particles isolated from HTLV-1-infected cells. These findings suggest that tRF-3019 may play an important role in priming HTLV-1 reverse transcription and could thus represent a novel target to control HTLV-1 infection. IMPORTANCE: Small noncoding RNAs, a growing family of regulatory RNAs that includes microRNAs and tRNA fragments, have recently emerged as key players in many biological processes, including viral infection and cancer. In the present study, we employed mass sequencing to identify the repertoire of small noncoding RNAs in normal T cells compared to T cells transformed with human T-cell leukemia virus type 1 (HTLV-1), a retrovirus that causes adult T-cell leukemia/lymphoma. The results revealed a distinct pattern of microRNA expression in HTLV-1-infected cells and a tRNA fragment (tRF-3019) that was packaged into virions and capable of priming HTLV-1 reverse transcription, a key event in the retroviral life cycle. These findings indicate tRF-3019 could represent a novel target for therapies aimed at controlling HTLV-1 infection.

Estimating the Severity of Obstructive Sleep Apnea Using ECG, Respiratory Effort and Neural Networks.

OBJECTIVE: wearable sensor technology has progressed significantly in the last decade, but its clinical usability for the assessment of obstructive sleep apnea (OSA) is limited by the lack of large and representative datasets simultaneously acquired with polysomnography (PSG). The objective of this study was to explore the use of cardiorespiratory signals common in standard PSGs which can be easily measured with wearable sensors, to estimate the severity of OSA. METHODS: an artificial neural network was developed for detecting sleep disordered breathing events using electrocardiogram (ECG) and respiratory effort. The network was combined with a previously developed cardiorespiratory sleep staging algorithm and evaluated in terms of sleep staging classification performance, apnea-hypopnea index (AHI) estimation, and OSA severity estimation against PSG on a cohort of 653 participants with a wide range of OSA severity. RESULTS: four-class sleep staging achieved a κ of 0.69 versus PSG, distinguishing wake, combined N1-N2, N3 and REM. AHI estimation achieved an intraclass correlation coefficient of 0.91, and high diagnostic performance for different OSA severity thresholds. CONCLUSIONS: this study highlights the potential of using cardiorespiratory signals to estimate OSA severity, even without the need for airflow or oxygen saturation (SpO2), traditionally used for assessing OSA. SIGNIFICANCE: while further research is required to translate these findings to practical and unobtrusive sensors, this study demonstrates how existing, large datasets can serve as a foundation for wearable systems for OSA monitoring. Ultimately, this approach could enable long-term assessment of sleep disordered breathing, facilitating new avenues for clinical research in this field.

Maximum a posteriori detection of heartbeats from a chest-worn accelerometer.

Objective. Unobtrusive long-term monitoring of cardiac parameters is important in a wide variety of clinical applications, such as the assesment of acute illness severity and unobtrusive sleep monitoring. Here we determined the accuracy and robustness of heartbeat detection by an accelerometer worn on the chest.Approach. We performed overnight recordings in 147 individuals (69 female, 78 male) referred to two sleep centers. Two methods for heartbeat detection in the acceleration signal were compared: one previously described approach, based on local periodicity, and a novel extended method incorporating maximumaposterioriestimation and a Markov decision process to approach an optimal solution.Main results. The maximumaposterioriestimation significantly improved performance, with a mean absolute error for the estimation of inter-beat intervals of only 3.5 ms, and 95% limits of agreement of -1.7 to +1.0 beats per minute for heartrate measurement. Performance held during posture changes and was only weakly affected by the presence of sleep disorders and demographic factors.Significance. The new method may enable the use of a chest-worn accelerometer in a variety of applications such as ambulatory sleep staging and in-patient monitoring.

Evidence-Based Lifestyle Guidelines and Self-Management Strategies Utilized by Women with Polycystic Ovary Syndrome.

Polycystic ovary syndrome (PCOS) is a complex endocrine disorder, affecting 13% of reproductive-aged women. While lifestyle management is the first-line treatment for improving complications, women experience challenges with implementation. This cross-sectional study aims to identify the types and sources of dietary and physical activity (PA) interventions implemented by women with PCOS and understand how they use self-management strategies to support lifestyle change. An online questionnaire was disseminated via a consumer-based PCOS website (May 2015-2016). Women (n = 1167) were aged 18-45 years and primarily born within the United States (70%). A quarter or less of women (diet 25%, PA 14%) sought lifestyle advice from health professionals (medical clinicians or dietitians) compared to over half (diet 59%, PA 67%) using alternative sources, namely from online platforms. While only 33% and 16% of women reported following formal dietary or PA guidelines, respectively, 57% had implemented a 'special diet' to manage their condition, many of which were inconsistent with evidence-based practice in PCOS. Participants also displayed a low level of engagement with important self-management behaviors, including goal setting and positive self-talk. These findings suggest that online information may promote inaccurate and ineffective lifestyle advice and emphasize the need to increase engagement with qualified health professionals.

Contactless Camera-Based Sleep Staging: The HealthBed Study.

Polysomnography (PSG) remains the gold standard for sleep monitoring but is obtrusive in nature. Advances in camera sensor technology and data analysis techniques enable contactless monitoring of heart rate variability (HRV). In turn, this may allow remote assessment of sleep stages, as different HRV metrics indirectly reflect the expression of sleep stages. We evaluated a camera-based remote photoplethysmography (PPG) setup to perform automated classification of sleep stages in near darkness. Based on the contactless measurement of pulse rate variability, we use a previously developed HRV-based algorithm for 3 and 4-class sleep stage classification. Performance was evaluated on data of 46 healthy participants obtained from simultaneous overnight recording of PSG and camera-based remote PPG. To validate the results and for benchmarking purposes, the same algorithm was used to classify sleep stages based on the corresponding ECG data. Compared to manually scored PSG, the remote PPG-based algorithm achieved moderate agreement on both 3 class (Wake-N1/N2/N3-REM) and 4 class (Wake-N1/N2-N3-REM) classification, with average κ of 0.58 and 0.49 and accuracy of 81% and 68%, respectively. This is in range with other performance metrics reported on sensing technologies for wearable sleep staging, showing the potential of video-based non-contact sleep staging.

Anti-VEGF therapy selects for clones resistant to glucose starvation in ovarian cancer xenografts.

BACKGROUND: Genetic and metabolic heterogeneity are well-known features of cancer and tumors can be viewed as an evolving mix of subclonal populations, subjected to selection driven by microenvironmental pressures or drug treatment. In previous studies, anti-VEGF therapy was found to elicit rewiring of tumor metabolism, causing marked alterations in glucose, lactate ad ATP levels in tumors. The aim of this study was to evaluate whether differences in the sensitivity to glucose starvation existed at the clonal level in ovarian cancer cells and to investigate the effects induced by anti-VEGF therapy on this phenotype by multi-omics analysis. METHODS: Clonal populations, obtained from both ovarian cancer cell lines (IGROV-1 and SKOV3) and tumor xenografts upon glucose deprivation, were defined as glucose deprivation resistant (GDR) or glucose deprivation sensitive (GDS) clones based on their in vitro behaviour. GDR and GDS clones were characterized using a multi-omics approach, including genetic, transcriptomic and metabolic analysis, and tested for their tumorigenic potential and reaction to anti-angiogenic therapy. RESULTS: Two clonal populations, GDR and GDS, with strikingly different viability following in vitro glucose starvation, were identified in ovarian cancer cell lines. GDR clones survived and overcame glucose starvation-induced stress by enhancing mitochondrial oxidative phosphorylation (OXPHOS) and both pyruvate and lipids uptake, whereas GDS clones were less able to adapt and died. Treatment of ovarian cancer xenografts with the anti-VEGF drug bevacizumab positively selected for GDR clones that disclosed increased tumorigenic properties in NOD/SCID mice. Remarkably, GDR clones were more sensitive than GDS clones to the mitochondrial respiratory chain complex I inhibitor metformin, thus suggesting a potential therapeutic strategy to target the OXPHOS-metabolic dependency of this subpopulation. CONCLUSION: A glucose-deprivation resistant population of ovarian cancer cells showing druggable OXPHOS-dependent metabolic traits is enriched in experimental tumors treated by anti-VEGF therapy.

Relationships between self-management strategies and physical activity and diet quality in women with polycystic ovary syndrome.

OBJECTIVE: To determine the relationships of self-management strategies and physical activity (PA) and diet quality in women with PCOS. METHODS: An online cross-sectional study involving women (n = 501), 18-45 years in the general Australian community with a self-reported PCOS diagnosis. The self-management and lifestyle behaviour questionnaires were completed between August 2017 and March 2018. RESULTS: Implementation of PA related self-management strategies increased the odds of meeting PA recommendations [Odds ratio (OR): 2.929 (95%CI: 2.172, 3.951), p < 0.001] but had no association with body mass index (BMI) [OR: 0.-0.984 (95%CI: -1.010, 0.959), p = 0.217] nor perception of self weight [OR: 1.382 (95% CI: 0.700, 2.725), p = 0.352]. Nutrition related self-management strategies were inversely associated with BMI [OR: - 0.115 (95%CI: -7.159, -0.980), p = 0.010] but had no association with diet quality [OR: 0.183 (95%CI: -2.328, 2.800), p = 0.855], energy intake [OR: - 0.092 (95%CI: -1204.443, 527.496) p = 0.438] or weight [OR: - 0.034 (95%CI: -4.020, 1.930), p = 0.491]. CONCLUSIONS: PA self-management strategies were associated with meeting PA recommendations. Nutrition strategies were associated with lower BMI but not diet quality, energy intake or weight in women with PCOS. PRACTICE IMPLICATIONS: Other behaviour change determinants (e.g. education, skills and self-efficacy) should be considered when designing a PCOS lifestyle programme in conjunction with self-management strategies.

Estimation of respiratory rate and effort from a chest-worn accelerometer using constrained and recursive principal component analysis.

Objective. Measurement of respiratory rate and effort is useful in various applications, such as the diagnosis of sleep apnea and early detection of patient deterioration in medical conditions, such as infections. A chest-worn accelerometer may be an easy and non-intrusive method, provided it is accurate and robust. We investigate the use of a novel method that can perform under realistic sleeping conditions such as variable sensor positions and body posture.Approach. Twenty subjects (aged 46-65 years) wore an accelerometer on the chest and a respiratory impedance plethysmography band as a reference. The subjects underwent an experimental protocol lasting approximately 90 min, under various postures and with different sensor positions. We used a novel, constrained, and recursive form of principal component analysis (PCA) to estimate the respiratory effort signal robustly. To obtain an estimate for the respiratory rate, first, multiple estimates were aggregated into a single frequency. Subsequently, a quality index was determined, such that unreliable estimates could be identified, and a trade-off could be made between coverage (percentage of time that the quality index is above a threshold) and limits of agreement.Main results. Results were determined over all recorded data, including changes in sensor position and posture. For respiratory effort, it was found that recursive and constrained computation of PCA reduced the estimation error significantly. For respiratory rate, a relation between coverage and limits of agreement was determined. If a minimum coverage of 80% was required, the limits of agreement could be kept below 1.45 breaths per minute. If the limits of agreement were constrained to 0.2 breaths per minute, a mean coverage of 5% was still attainable.Significance. We have shown that chest-worn accelerometery can be a robust and accurate method for measurement of respiratory features under realistic conditions.

Genetic Perturbation of Pyruvate Dehydrogenase Kinase 1 Modulates Growth, Angiogenesis and Metabolic Pathways in Ovarian Cancer Xenografts.

Pyruvate dehydrogenase kinase 1 (PDK1) blockade triggers are well characterized in vitro metabolic alterations in cancer cells, including reduced glycolysis and increased glucose oxidation. Here, by gene expression profiling and digital pathology-mediated quantification of in situ markers in tumors, we investigated effects of PDK1 silencing on growth, angiogenesis and metabolic features of tumor xenografts formed by highly glycolytic OC316 and OVCAR3 ovarian cancer cells. Notably, at variance with the moderate antiproliferative effects observed in vitro, we found a dramatic negative impact of PDK1 silencing on tumor growth. These findings were associated with reduced angiogenesis and increased necrosis in the OC316 and OVCAR3 tumor models, respectively. Analysis of viable tumor areas uncovered increased proliferation as well as increased apoptosis in PDK1-silenced OVCAR3 tumors. Moreover, RNA profiling disclosed increased glucose catabolic pathways-comprising both oxidative phosphorylation and glycolysis-in PDK1-silenced OVCAR3 tumors, in line with the high mitotic activity detected in the viable rim of these tumors. Altogether, our findings add new evidence in support of a link between tumor metabolism and angiogenesis and remark on the importance of investigating net effects of modulations of metabolic pathways in the context of the tumor microenvironment.

Corrigendum to "Genetic perturbation of IFN-α transcriptional modulators in human endothelial cells uncovers pivotal regulators of angiogenesis" [Comput Struct Biotechnol J. 2020 Dec 2;18:3977-3986. doi: https://doi.org//10.1016/j.csbj.2020.11.048. eCollection 2020].

[This corrects the article DOI: 10.1016/j.csbj.2020.11.048.].

A Markov chain probability model of glucose tolerance in post gestational diabetes follow up study.

Women with gestational diabetes mellitus (GDM) are at increased risk of developing type 2 diabetes (T2DM). However, the degree of risk and the timing of progression from normal to a pre-diabetic or diabetic state have not been clearly quantified. In this study we analyzed data from a longitudinal study on a group of women with a history of GDM, that were inserted in an oral glucose tolerance test (OGTT) annual screening program and followed up for 5 years after partum. A three state Markov chain model was proposed to represent the dynamics of changes between metabolic states. We used the data to empirically estimate the one-year transition parameters of the model and make predictions about the possibility that women with normal glucose tolerance or impaired glucose metabolism just after partum will develop overt T2DM in three or five years. Results show that subjects with an impaired glucose metabolism few months after partum will hardly (10%) be in the same state after three years. Women with normal glucose tolerance after partum will have a high probability (0.80) to be in the same state three years after.