Endogenous Control Research Articles

Survey of Xylella fastidiosa Infection in Prunus Germplasm in Gainesville, FL, USA

The pathogen Xylella fastidiosa is a xylem-restricted, gram-negative bacterium that is known to cause diseases of many cultivated plant species. Recent outbreaks of X. fastidiosa diseases in Europe have brought attention to the impact of this pathogen, especially to perennial crops. Among the Prunus genus, X. fastidiosa is known to have a wide range of hosts, including plum, almond, peach, cherry, and apricot. Infected trees have reduced fruit quality, possibly resulting in unmarketable fruits, as well as reduced lifespan. There are no curative management options for X. fastidiosa diseases in Prunus; therefore, development of resistant or tolerant cultivars through breeding represents an efficient option to reduce the impact of this pathogen. In this context, the main objective of this study was to determine the occurrence of X. fastidiosa in germplasm of the Stone Fruit Breeding Program at the University of Florida located in Gainesville, FL, USA, under natural infection conditions. A total of 43 individuals representing 10 different genotypic groups within the Prunus genus were tested for the presence of X. fastidiosa. Additionally, we report a novel and easy sampling method using sawdust collected from tree trunks for the detection of this pathogen in Prunus and the development of an endogenous control for improving the diagnosis of this pathogen using real-time polymerase chain reactions. Our results showed a high incidence of X. fastidiosa in the germplasm tested, with more than 65% of the samples positive for the presence of the bacterial pathogen. However, X. fastidiosa was not detected in most of the P. mume samples tested, whereas almost all the P. mume × P. armeniaca hybrids were positive. Negative individuals were also identified in P. avium, P. campanulata, P. umbellata, and P. salicina × P. ceracifera. These trees have been planted in the field, exposed to natural infection for 4 to 11 years, and are considered to show field resistance. Finally, primers and probes based on the Prunus COX gene developed in this study can be used as an internal amplification control to enhance the interpretation of results of X. fastidiosa detection assays using sawdust samples.

Anastomotic stenosis of bioengineered trachea grafts is driven by transforming growth factor β1-induced signaling, proinflammatory macrophages, and delayed epithelialization

ObjectiveAnastomotic stenosis caused by hypertrophic granulation tissue often develops in response to orthotopically implanted bioengineered tracheal grafts. To determine mechanisms responsible for the development and persistence of this granulation tissue, we looked for changes in gene expression from tissue specimens from the graft-native interface. MethodsRNA was isolated from paraffin-embedded tissue samples of the anastomotic sites of orthotopically implanted bioengineered tracheal grafts of 9 animals. Tissue samples were binned into 3 groups based on degree of stenosis: no stenosis (<5%), mild stenosis (25%-50%), and moderate and severe stenosis (≥75%). Sections of healthy trachea tissue were used as control. The expression levels of ∼200 genes related to wound healing, plus several endogenous controls, were measured with a pathway-focused predesigned primer array. ResultsExpression of ARG2, IL4, RPL13 A, TGFBR3, and EGFR decreased, whereas expression of RUNX2 was increased in stenotic wounds compared with nonstenotic tissue. Based on the cell types present in the trachea and wound healing, this expression profile indicates a lack of M2 anti-inflammatory macrophages, absent epithelial cells, and transforming growth factor β1-induced signaling. ConclusionsThese findings represent a significant step for tracheal tissue engineering by identifying several key mechanisms present in stenotic granulation tissue. Further research must be conducted to determine what modifications of the graft substrate and which coadministered therapeutics can be used to prevent the development of hypertrophic granulation tissue.

Evolution of molecular switches for regulation of transgene expression by clinically licensed gluconate.

Synthetic biology holds great promise to improve the safety and efficacy of future gene and engineered cell therapies by providing new means of endogenous or exogenous control of the embedded therapeutic programs. Here, we focused on gluconate as a clinically licensed small-molecule inducer and engineered gluconate-sensitive molecular switches to regulate transgene expression in human cell cultures and in mice. Several switch designs were assembled based on the gluconate-responsive transcriptional repressor GntR from Escherichia coli. Initially we assembled OFF- and ON-type switches by rewiring the native gluconate-dependent binding of GntR to target DNA sequences in mammalian cells. Then, we utilized the ability of GntR to dimerize in the presence of gluconate to activate gene expression from a split transcriptional activator. By means of random mutagenesis of GntR combined with phenotypic screening, we identified variants that significantly enhanced the functionality of the genetic devices, enabling the construction of robust two-input logic gates. We also demonstrated the potential utility of the synthetic switch in two in vivo settings, one employing implantation of alginate-encapsulated engineered cells and the other involving modification of host cells by DNA delivery. Then, as proof-of-concept, the gluconate-actuated genetic switch was connected to insulin secretion, and the components encoding gluconate-induced insulin production were introduced intotype-1 diabetic miceas naked DNA via hydrodynamic tail vein injection. Normoglycemia was restored, thereby showcasing the suitability of oral gluconate to regulate in situ production of a therapeutic protein.

Identification and expression pattern of aluminium-responsive genes in roots of rice genotype with reference to Al-sensitivity

Aluminium (Al) is the third most abundant element in the Earth's crust. Globally, acidic soil occupies 30–40% of ice-free land areas; Al toxicity is a major threat to crops. The first symptom of Al toxicity is the inhibition of root growth followed by poor root hair development, swollen root apices, necrosis of leaves and reduced yield. Although Rice (Oryza sativa) is an Al toxicity tolerant crop, it shows considerable variations among rice genotypes to Al exposure. Therefore, it is pertinent to understand Al toxicity and underlying mechanisms for Al tolerance in Rice. In the present study, 63 rice genotypes screened under Al stress showed significant variations of root growth. Expression stability of endogenous control genes (ECGs) revealed sulphite reductase (SR) as the most stable ECG that can be used as a reference gene for quantitative real-time PCR (qRT-PCR). Expression patterns of Al-responsive genes suggest genes associated with cytoskeletal dynamics, metabolism, and ion transporter could play significant roles in Al adaptation and tolerance in rice. The results showed Motodhan, Vietnam-1, Yimyu and N-861 as Al-toxicity tolerant, while Lespah, RCPL-13, VL-31329, and UPR2919-141-1 as most Al-sensitive genotypes among the studied rice lines cultivated in North-East India.

The microRNA-144/451 cluster in plasma-derived microvesicles and erythrocytes in patients with history of pulmonary embolism

Chronic thromboembolic disease (CTED) and chronic thromboembolic pulmonary hypertension (CTEPH) are the complications that comprise a serious problem for patients with history of pulmonary embolism (PE). Erythrocytes, extracellular microvesicles (EMVs) and miRNAs play a substantial role in the procoagulant states. The aim. To analyze the levels of miR-144-3р, miR-451a, and miR-451b in blood plasma-derived EMVs and erythrocytes in patients with history of PE and in the control group. Materials and Methods. 18 patients with history of PE (13 CTEPH, 5 CTED) and 8 controls were enrolled into the study. All the participants had undergone clinical and biochemical blood tests as well as the coagulogram. We used flow cytometry to assess plasma-derived EMVs (CD9, CD41, CD45, CD235a, CD105). We measured the expression of miR-144-3р, miR-451a, miR-451b by real-time PCR with endogenous control (miR-152-3p) and five exogenous quality controls. Results. The levels of miR-144-3р and miR-451a in patients were lower than in controls, both in EMVs (р = 0.030; р = 0.065) and in erythrocytes (р = 0.023;р = 0.086). In female patients, the levels of miR-144-3р and miR-451a in CTEPH were lower than in CTED (р = 0.087; р = 0.031). Mir-451b in EMVs has not been detected, while in erythrocytes its levels have not differed between the groups. In patients, the levels of miR-144-3р and miR-451a directly correlated with each other both in EMVs (р = 0.004) and in erythrocytes (р = 0.042). In all the participants, the levels of miR-144-3р and miR-451a in EMVs directly correlated with those in erythrocytes (р = 0.002; р = 0.078). The number of erythrocyte-derived EMVs correlated with miR-451a levels both in EMVs (R = 0.472; p = 0.065) and in erythrocytes (R = –0.829; p = 0.011). The level of miR-451a in EMVs correlated with blood plasma levels of factor VIII and fibrinogen (R = 0.584; p = 0.022 and R= –0.489; p = 0.047), and with the International Normalized Ratio (R = 0.894; p = 0.041). Conclusion. The microRNA-144/451 cluster may influence both the hemostasis system and the risk of post-thromboembolic complications development. In the present study, miR-144-3р and miR-451a showed themselves as protective factors in relation to both the development of PE and severity of post-thromboembolic complications.

POSTСOLITIS ALTERATIONS IN NOCICEPTIVE PROPERTIES OF NEURONS IN THE RAT RAPHE MAGNUS AND DORSAL RAPHE NUCLEI

Dysfunction of the brain serotonergic system is thought to play a leading role in the pathogenesis of chronic abdominal pain and comorbid somatic hyperalgesia, which disturb a significant proportion of patients with digestive tract diseases, even in remission. However, the specific changes in the serotonergic structures nociceptive properties that can be initiated by organic pathology and persist after its resolution remain unclear. The aim of our neurophysiological study on anesthetized rats – healthy and recovered from colitis – was to identify the alterations in the raphe magnus (RMg) and dorsal raphe (DR) nuclei neuronal responses to visceral (colorectal distension) and somatic (squeezing of the tail) noxious stimulations that persist after resolution of intestinal inflammation. It has been shown that both nuclei contain different groups of nociceptive neurons: 1) responding with activation only to colorectal distension (visceral); 2) excited only by tail squeezing (somatic); 3) reacting with excitation to the both irritations (general); 4) responding with discharge inhibition to any of the stimulations (inhibited). Compared with healthy animals, in RMg of colitis-exposed rats the number of the inhibited cells was increased and the total proportion of excited nociceptive neurons was reduced. Distension of the inflammation-undergone intestine caused enhanced RMg neuronal inhibition, whereas squeezing of the pathology-unaffected tail led to increased excitation of the RMg selective somatic and general nociceptive cells. In turn, in the DR of postcolitis rats the inhibited neuron proportion was reduced, while the increased population of excited neurons included fewer visceral and more somatic selective cells. This was accompanied by an increase in the selective reactions of the latter to somatic pain stimuli and by an increase in non-selective DR neuron excitation by visceral and somatic pain signals. The identified neuronal alterations can contribute to the postcolitis impairment of the studied raphe nuclei functions in the endogenous control of visceral and somatic pain sensitivity.

Spring arrival of the common cuckoo at breeding grounds is strongly determined by environmental conditions in tropical Africa.

Failure to adapt migration timing to changes in environmental conditions along migration routes and at breeding locations can result in mismatches across trophic levels, as occurs between the brood parasitic common cuckoo Cuculus canorus and its hosts. Using satellite tracking data from 87 male cuckoos across 11 years, we evaluate why the cuckoo has not advanced its arrival to the UK. Across years, breeding ground arrival was primarily determined by timing of departure from stopover in West Africa before northward crossing of the Sahara. Together with high population synchrony and low apparent endogenous control of this event, this suggests that a seasonal ecological constraint operating here limits overall variation in breeding grounds arrival, although this event was itself influenced by carry-over from timing of arrival into tropical Africa. Between-year variation within individuals was, in contrast, mostly determined by northward migration through Europe, probably due to weather conditions. We find evidence of increased mortality risk for (a) early birds following migration periods positively impacting breeding grounds arrival, and (b) late birds, possibly suffering energy limitation, after departure from the breeding grounds. These results help identify areas where demands of responding to global change can potentially be alleviated by improving stopover quality.

Prolactin reduces the kainic acid-induced increase in intracellular Ca2+ concentration, leading to neuroprotection of hippocampal neurons

The aim of this study was to determine the effect of prolactin (PRL) on intracellular calcium (Ca2+) concentration and its neuroprotective role in a model of kainic acid (KA) excitotoxicity in primary cultures of hippocampal neurons. Cell viability and intracellular Ca2+ concentrations were determined by MTT and Fura-2 assays, respectively, either after induction by KA as an agonist or after treatment with NBQX antagonist alone or in combination with PRL administration. Expression of ionotropic glutamatergic receptors (iGluRs) subunits in neuronal cells was determined by RT-qPCR. Dose-response treatments with KA or glutamate (Glu), the latter used as endogenous agonist control, induced a significant increase in neuronal intracellular Ca2+ concentration followed by a significant decrease in hippocampal neuronal viability. Administration of PRL induced a significant increase in neuronal viability after treatment with KA. Furthermore, administration of PRL decreased intracellular Ca2+ concentrations induced by KA treatment. Independent administration of the AMPAR-KAR antagonist reversed cell death and reduced intracellular Ca2+ concentration in a similar manner as PRL. Additionally, mRNA expression of AMPAR, KAR and NMDAR subtypes were detected in hippocampal neurons; however, no significant changes in iGluRs subunit expression were observed due to excitotoxicity or PRL treatment. The results suggest that PRL inhibits the increase in intracellular Ca2+ concentration induced by KA, leading to neuroprotection.

Sap flow responses in pioneer and late-successional tree species in secondary tropical montane forests of Eastern Himalaya, India.

The interactive role of life-history traits and environmental forcing on plant-water relations is crucial for understanding species response to climate change but remains poorly understood in secondary tropical montane forests (TMFs). Comparing contrasting life-history traits (pioneer vs late-successional species) in a biodiverse Eastern Himalayan secondary TMF, we investigated sap flow responses in co-occurring pioneer species, Symplocos racemosa (n=5) and Eurya acuminata (n=5), and late-successional species, Castanopsis hystrix (n=3), using modified Granier's Thermal Dissipation probes. The fast-growing pioneers S. racemosa and E. acuminata) had 2.1- and 1.6-times higher sap flux density than the late-successional C. hystrix, respectively, and exhibited characteristics of long-lived pioneer species. Significant radial and azimuthal variability in sap flow (V) between species was observed and attributed to life history traits and the canopy's access to sunlight. Nocturnal V (1800-0500hr) was 13.8 % of daily V and is attributed to stem recharge for evening V (1800-2300hr) and to endogenous stomatal controls for pre-dawn V (0000-0500hr). Both the shallow-rooted pioneer species exhibited midday depression in V attributed to photosensitivity and diel moisture stress response. In contrast, deep-rooted C. hystrix transpired unaffected across the dry season likely accessing groundwater. Thus, the secondary broadleaved TMFs, with the dominance of shallow-rooted pioneers, are more prone to the negative impacts of drier and warmer winters than primary forests, which are dominated by deep-rooted species. The study provides an empirical understanding of life-history traits and microclimate modulating plant-water use in widely distributed secondary TMFs in Eastern Himalaya and highlights their vulnerability against warmer winters and reduced snowfall due to climate change.

Effect of Voluntary Exercise on Endogenous Pain Control Systems and Post-traumatic Headache in Mice

Traumatic brain injury (TBI) can cause acute and chronic pain along with motor, cognitive, and emotional problems. Although the mechanisms are poorly understood, previous studies suggest disruptions in endogenous pain modulation may be involved. Voluntary exercise after a TBI has been shown to reduce some consequences of injury including cognitive impairment. We hypothesized, therefore, that voluntary exercise could augment endogenous pain control systems in a rodent model of TBI. For these studies, we used a closed-head impact procedure in male mice modeling mild TBI. We investigated the effect of voluntary exercise on TBI-induced hindpaw nociceptive sensitization, diffuse noxious inhibitory control failure, and periorbital sensitization after bright light stress, a model of post-traumatic headache. Furthermore, we investigated the effects of exercise on memory, circulating markers of brain injury, neuroinflammation, and spinal cord gene expression. We observed that exercise significantly reduced TBI-induced hindpaw allodynia and periorbital allodynia in the first week following TBI. We also showed that exercise improved the deficits associated with diffuse noxious inhibitory control and reduced bright light stress-induced allodynia up to 2 months after TBI. In addition, exercise preserved memory and reduced TBI-induced increases in spinal BDNF, CXCL1, CXCL2, and prodynorphin expression, all genes previously linked to TBI-induced nociceptive sensitization. Taken together, our observations suggest that voluntary exercise may reduce pain after TBI by reducing TBI-induced changes in nociceptive signaling and preserving endogenous pain control systems. PerspectiveThis article evaluates the effects of exercise on pain-related behaviors in a preclinical model of traumatic brain injury (TBI). The findings show that exercise reduces nociceptive sensitization, loss of diffuse noxious inhibitory control, memory deficits, and spinal nociception-related gene expression after TBI. Exercise may reduce or prevent pain after TBI.

The pennate diatom Pseudo-nitzschia multistriata as a model for diatom life cycles, from the laboratory to the sea.

Phytoplankton dynamics are regulated by external cues, such as light and nutrients, as well as by biotic interactions and endogenous controls linked to life cycle characteristics. The planktonic pennate diatom Pseudo-nitzschia multistriata, with a heterothallic mating system with two opposite mating types (MTs), represents a model for the study of diatom life cycles. P. multistriata is a toxic species, able to produce the neurotoxin domoic acid. First described in Japan in 1993, it was detected at the long-term monitoring station MareChiara (Gulf of Naples, Italy) in 1995. Since then, P. multistriata has been reported from several worldwide coastal sites. A large body of knowledge has been produced on its ecology, genetic diversity, and life cycle characteristics. The availability of these data, the ecological relevance of the Pseudo-nitzschia genus, and its controllable life cycle with a short generation time made it an ideal species to develop a genetic model system for diatoms. To enable functional studies, a 59 Mb genome sequence and several transcriptomic data were produced, and genetic transformation was optimized. These tools allowed the discovery of the first mating-type determining gene for diatoms. Gene expression studies and metabolomics analyses defined genes and molecules underpinning different phases of the process of sexual reproduction. This model system, developed to explore the genetics of diatom life cycles, offers the opportunity to parallel experimental observations in the laboratory using in situ meta-omics analyses along space and time, empowering knowledge on the biology and ecology of the genus.

The start-up's roadmap to private equity financing: substituting discounts with a premium in valuation for growth

PurposeAccess to financing is a key success factor for start-ups. High failure rates, long payback periodse and asymmetries lead to conservative pricing and valuation discounts. The authors examine financial marketing and contingent factors, as enablers of a “patent premium” by private equity (PE) investors targeting start-ups in their growth and expansion stages.Design/methodology/approachDrawing from the contingency, innovation and signaling theories, the authors collect patent records for Italian start-ups in which a higher than 30% stake was acquired by PE investors during the period 2014–2020. The authors apply a generalized linear model with a logit link and robust clustered error to test the key relationships and control for endogeneity with a Heckman two-stage selection model.FindingsFindings indicate start-ups’ access to financing is significantly impacted by marketing constructs adopted in the operation. Innovation alone does not suffice to determine a valuation premium, unless contingent on the promotion of its product, the placement -investors targeted-of the equity, brand equity levers of previous ownership and marketing competence backing the deal.Originality/valueThe authors provide new insights in the marketing-finance interface, highlighting levers that reassure investors and enable monetizing innovation in start-ups that are still privately held. The authors bridge a gap in literature that has mainly focused on venture capital and innovation financing in the open market, as well as a significant gap regarding the marketing design of private equity placements.

Do foreign institutional investors influence corporate climate change disclosure quality? International evidence

AbstractResearch Question/IssueWe examine the association between foreign institutional ownership and climate change disclosure quality from 2006 to 2018 across 34 countries. We find that firms with a higher level of foreign institutional ownership demonstrate better quality climate change disclosures, whereas domestic institutional ownership has immaterial impacts on such disclosures. We utilize a difference‐in‐differences (DiD) analysis using a firm's addition to the Morgan Stanley Capital International (MSCI) index as an exogenous shock to control for endogeneity. Our findings are robust to various other endogeneity controls. We also establish evidence on an indirect effect of climate change disclosure quality in mediating the positive association between foreign institutional investors and firm valuation.Research Findings/InsightsWe find that the positive association between foreign institutional ownership and climate change disclosure quality is more pronounced for (1) firms domiciled in stakeholder‐orientated countries, (2) firms domiciled in countries that adopt emission trading schemes, and (3) firms with a greater level of information asymmetry. Additionally, our results are more robust when foreign investors are domiciled in countries that care more about the environment.Theoretical/Academic ImplicationsOur study contributes to climate change disclosures, corporate governance, and international business literature by showing that foreign rather than domestic institutional investors contribute to improved corporate climate change disclosure quality in their portfolio firms.Practitioner/Policy ImplicationsOur study urges regulators to increase their market oversight, especially in firms with less foreign institutional ownership. This is required because such firms are prone to exhibiting poorer accountability for their climate risk management practices, and their disclosures are bereft of effective external monitoring mechanisms.

Small Business Property Tax Reductions and Firm Productivity

We investigate the productivity spillovers from the UK government’s decision to use extensive property tax reductions as a key instrument to stimulate innovation in smaller businesses and drive local growth. To capture the complex interaction and clustering of hierarchical effects, we apply non-parametric Random Effects Expectation Maximisation algorithm that complements more standard econometric estimators, namely matching to control for endogeneity and control functions to estimate total factor productivity. These approaches enabled us to incorporate various contextual configurations in comparing the recipients of these reductions to non-recipients with regard to productivity, in which the UK has experienced a considerably worse performance than its peers since the great recession. Contrary to policy assumptions and business community expectations, we show that generic tax reductions, when significant, are mostly associated with lower productivity and thus have been unsuitably chosen as a policy mechanism to stimulate productivity growth. We further show how instruments that are not built for causality could be beneficial for policy evaluation.

Strategies for data normalization and missing data imputation and consequences for potential diagnostic microRNA biomarkers in epithelial ovarian cancer.

MicroRNAs (miRNAs) are small non-coding RNA molecules regulating gene expression with diagnostic potential in different diseases, including epithelial ovarian carcinomas (EOC). As only a few studies have been published on the identification of stable endogenous miRNA in EOC, there is no consensus which miRNAs should be used aiming standardization. Currently, U6-snRNA is widely adopted as a normalization control in RT-qPCR when investigating miRNAs in EOC; despite its variable expression across cancers being reported. Therefore, our goal was to compare different missing data and normalization approaches to investigate their impact on the choice of stable endogenous controls and subsequent survival analysis while performing expression analysis of miRNAs by RT-qPCR in most frequent subtype of EOC: high-grade serous carcinoma (HGSC). 40 miRNAs were included based on their potential as stable endogenous controls or as biomarkers in EOC. Following RNA extraction from formalin-fixed paraffin embedded tissues from 63 HGSC patients, RT-qPCR was performed with a custom panel covering 40 target miRNAs and 8 controls. The raw data was analyzed by applying various strategies regarding choosing stable endogenous controls (geNorm, BestKeeper, NormFinder, the comparative ΔCt method and RefFinder), missing data (single/multiple imputation), and normalization (endogenous miRNA controls, U6-snRNA or global mean). Based on our study, we propose hsa-miR-23a-3p and hsa-miR-193a-5p, but not U6-snRNA as endogenous controls in HGSC patients. Our findings are validated in two external cohorts retrieved from the NCBI Gene Expression Omnibus database. We present that the outcome of stability analysis depends on the histological composition of the cohort, and it might suggest unique pattern of miRNA stability profiles for each subtype of EOC. Moreover, our data demonstrates the challenge of miRNA data analysis by presenting various outcomes from normalization and missing data imputation strategies on survival analysis.

Influence of the environment on the early development of attentional control

The control of visual attention is key to learning and has a foundational role in the development of self-regulated behavior. Basic attention control skills emerge early in life and show a protracted development along childhood. Prior research suggests that attentional development is influenced by environmental factors in early and late childhood. Although, much less information is available about the impact of the early environment on emerging endogenous attention skills during infancy. In the current study we aimed to test the impact of parental socioeconomic status (SES) and home environment (chaos) in the emerging control of orienting in a sample of typically-developing infants. A group of 142 (73 female) 6-month-old infants were longitudinally tested at 6, 9 (n = 122; 60 female) and 16–18 (n = 91; 50 female) months of age using the gap-overlap paradigm. Median saccade latency (mdSL) and disengagement failure (DF) were computed as dependent variables for both overlap and gap conditions. Also, composite scores for a Disengagement Cost Index (DCI) and Disengagement Failure Index (DFI) were computed considering mdSL and DF of each condition, respectively. Families reported SES and chaos in the first and last follow-up sessions. Using Linear Mixed Models with Maximum Likelihood estimation (ML) we found a longitudinal decrease in mdSL in the gap but not in the overlap condition, while DF decreased with age independently of the experimental condition. Concerning early environmental factors, an SES index, parental occupation and chaos at 6 months were found to show a negative correlation with DFI at 16–18 months, although in the former case it was only marginally significant. Hierarchical regression models implementing ML showed that both SES and chaos at 6 months significantly predicted a lower DFI at 16–18 months. Results show a longitudinal progression of endogenous orienting between infancy and toddlerhood. With age, an increased endogenous control of orienting is displayed in contexts where visual disengagement is facilitated. Visual orienting involving attention disengagement in contexts of visual competition do not show changes with age. Moreover, these attentional mechanisms of endogenous control seem to be modulated by early experiences of the individual with the environment.

What to Do if the qPCR Test for SARS-CoV-2 or Other Pathogen Lacks Endogenous Internal Control? A Simple Test on Housekeeping Genes.

Some of the products for the molecular diagnosis of infections do not have an endogenous internal control, and this is necessary to ensure that the result is not a false negative. The aim of the project was to design a simple low-cost RT-qPCR test that can confirm the expression of basic metabolism proteins, thus confirming the quality of genetic material for molecular diagnostic tests. Two successful equivalent qPCR assays for the detection of the GADPH and ACTB genes were obtained. The course of standard curves is logarithmic, with a very high correlation coefficient R2 within the range of 0.9955-0.9956. The reaction yield was between 85.5 and 109.7%, and the detection limit (LOD) with 95% positive probability was estimated at 0.0057 ng/µL for GAPDH and 0.0036 ng/µL for ACTB. These tests are universal because they function on various types of samples (swabs, cytology, etc.) and can complement the diagnosis of SARS-CoV-2 and other pathogens, as well as potentially oncological diagnostics.

The Role of Graphene Oxide in the Inhibition of Migration and Invasion of Cancer Cells by Destroying Actin Cytoskeleton and via Inhibiting the Activities of ETC Complexes

Abstract: Cancer is one of the most important threats to public health. Cancer is characterized by cell proliferation that has eluded central endogenous control mechanisms. Cervical cancer is the third most common cancer among women, followed by skin cancer and breast cancer, the first and second most common causes, respectively. In developing countries, cervical cancer is usually the most common cancer in women and may account for 25% of all female cancers. Over the years, the diagnosis and treatment of cervical cancer have made rapid progress, resulting in a decline in the mortality and morbidity of the disease. Unfortunately, although surgery and radiotherapy have effective treatment options for early cervical cancer, poor prognosis is still a challenge in the treatment of metastatic cervical cancer. Therefore, it is very important to reveal the mechanism of cervical cancer and explore new therapies against tumor invasiveness. At present, it is reported that nanomaterials are used in the detection and treatment of various types of malignant tumors due to their different targeting effects in many fields, such as imaging, immune detection, chemotherapy, radiotherapy and immunotherapy. The cytotoxicity and inhibitory effects of graphene oxide (GO) on tumor invasion and metastasis were studied in recent studies using the human cervical cancer Hela cell line, as well as the probable mechanisms and signaling pathways involved. Here we collect the last reports, with focus on the role of GO in the inhibition of migration and invasion of cancer cells with the goal of exposing possible potential pathways to provide new insights for specific treatment of cancer.

How does the quality of care for type 2 diabetic patients benefit from GPs-nurses' teamwork? A staggered difference-in-differences design based on a French pilot program.

In many countries, policies have explicitly encouraged primary care teams and inter-professional cooperation and skill mix, as a way to improve both productive efficiency gains and quality improvement. France faces barriers to developing team working as well as new and more advanced roles for health care professionals including nurses. We aim to estimate the impact of a national pilot experiment of teamwork between general practitioners (GPs) and advance practitioners nurses (APN)-who substitute and complement GPs-on yearly quality of care process indicators for type two diabetes patients (T2DP). Implemented by a not-for-profit meso-tier organisation and supported by the Ministry of Health, the pilot relied on the voluntary enrolment of newly GPs from 2012 to 2015; the staffing and training of APNs; skill mixing and new remuneration schemes. We use latent-response formulation models, control for endogeneity and selection bias by using controlled before-after and quasi-experimental design combining coarsened exact matching-prior to the treatment, at both GPs (435 treated vs 973 control) and T2DP levels -, with intention to treat (ITT; 18,310 in each group) and per protocol (PP, 2943 in each group) perspectives, as well as difference-in-differences estimates on balanced panel claims data from the National Health Insurance Fund linked to clinical data over the period 2010-2017. We show evidence of a positive and significant positive impact for T2DP followed-up by newly enrolled GPs in the pilot compared to the pretreatment period and the control group. The effect magnitudes were larger for PP than for ITT subsamples.

Selection of endogenous control and identification of significant microRNA deregulations in cervical cancer.

Cervical cancer causes approximately 350,000 deaths each year. The availability of sensitive and specific diagnostic tests to detect cervical cancer in its early stages is essential to improve survival rates. In this study, we compared two strategies for selecting endogenous controls: miRNA profiling by small-RNA sequencing and a commercially available microfluidic card with 30 recommended endogenous controls preloaded by the manufacturer. We used the RefFinder algorithm and coefficient of variation to select endogenous controls. We selected the combination of miR-181a-5p and miR-423-3p as the most optimal normalizer. In the second part of this study, we determined the differential expression (between tumor/non-tumor groups) of microRNA in cervical cancer FFPE tissue samples. We determined the comprehensive miRNA expression profile using small-RNA sequencing technology and verified the results by real-time PCR. We determined the relative expression of selected miRNAs using the 2-ΔΔCt method. We detected statistically significant upregulation of miR-320a-3p, miR-7704, and downregulation of miR-26a-5p in the tumor group compared to the control group. The combination of these miRNAs may have the potential to be utilized as a diagnostic panel for cervical cancer. Using ROC curve analysis, the proposed panel showed 93.33% specificity and 96.97% sensitivity with AUC = 0.985. We proposed a combination of miR-181a-5p and miR-423-3p as optimal endogenous control and detected potentially significant miRNAs (miR-320a-3p, miR-7704, miR-26a-5p). After further validation of our results, these miRNAs could be used in a diagnostic panel for cervical cancer.