Far Red-Shifted CdTe Quantum Dots for Multicolour Stimulated Emission Depletion Nanoscopy.

Stimulated emission depletion (STED) nanoscopy is a widely used nanoscopy technique. Two-colour STED imaging in fixed and living cells is standardised today utilising both fluorescent dyes and fluorescent proteins. Solutions to image additional colours have been demonstrated using spectral unmixing, photobleaching steps, or long-Stokes-shift dyes. However, these approaches often compromise speed, spatial resolution, and image quality, and increase complexity. Here, we present multicolour STED nanoscopy with far red-shifted semiconductor CdTe quantum dots (QDs). STED imaging of the QDs is optimized to minimize blinking effects and maximize the number of detected photons. The far-red and compact emission spectra of the investigated QDs free spectral space for the simultaneous use of fluorescent dyes, enabling straightforward three-colour STED imaging with a single depletion beam. We use our method to study the internalization of QDs in cells, opening up the way for future super-resolution studies of particle uptake and internalization.

Hadamard-transform spectral acquisition with an acousto-optic tunable filter in a broadband stimulated Raman scattering microscope.

We present a novel configuration for high spectral resolution multiplexing acquisition based on the Hadamard transform in stimulated Raman scattering (SRS) microscopy. The broadband tunable output of a dual-beam femtosecond laser is filtered by a fast, narrowband, and multi-channel acousto-optic tunable filter (AOTF). By turning on and off different subsets of its 8 independent channels, the AOTF generates the spectral masks given by the Hadamard matrix. We demonstrate a seamless and automated operation in the Raman fingerprint and CH-stretch regions. In the presence of additive noise, the spectral measurements using the multiplexed method show the same signal-to-noise ratio of conventional single-wavenumber acquisitions performed with 4 times longer integration time.

Dental age estimation in children affected by juvenile rheumatoid arthritis.

Dental root calcification has proven to be a reliable biological evidence to estimate chronological age of children. The development of structures usually examined in the age estimation forensic practice (e.g. skeleton, teeth) is supposed to be influenced by diseases and nutritional, environmental, ethnic, and ultimately even socioeconomic factors. This research aims to study the age estimation in children affected by juvenile rheumatoid arthritis (JRA) with and without steroids treatment and compared with healthy subjects. MATERIAL AND METHODS: Dental age estimations based on 752 OPGs, 420 girls and 332 boys, aged from 3.3 to 15.99 years, were provided by applying Demirjian and Willems' original methods. Of the whole sample, 103 individuals were affected by JRA and 40 received a continuous corticosteroid therapy, over 1 year long. CONCLUSIONS: Willems' and Demirjian's original methods, as methods commonly applied to estimate age for sub-adults with unremarkable medical history, can be used for medico-legal purposes to children affected by JRA. Willems' method tended to underestimate age while Demirjian's method resulted to be prone to overestimation for both healthy and JRA-affected children. JRA showed to have no influence on root calcification process even in children that received steroid treatment for 1 year or longer.

Predictors of Severe Neurologic Injury on Ultrasound Scan of the Head and Risk Factor-based Screening for Infants Born Preterm.

OBJECTIVE: To identify risk factors for severe neurologic injury (intraventricular hemorrhage grade 3 or greater and/or periventricular leukomalacia) diagnosed by ultrasound scan of the head among infants born at 300-326 weeks of gestation and compare different screening strategies. STUDY DESIGN: This was a retrospective cohort study of infants born at 300-326 weeks or >326 weeks of gestation with a birth weight <1500 g admitted to neonatal intensive care units in the Canadian Neonatal Network from 2011 to 2016. Stepwise logistic regression analysis was used to identify significant risk factors and calculate aORs and 95% CIs. Risk factor-based screening strategies were compared. RESULTS: The rate of severe neurologic injury was 3.1% among infants screened (285/9221). Significant risk factors included singleton birth (aOR 1.96, 95% CI 1.35-2.85), 5-minute Apgar <7 (aOR 1.81, 95% CI 1.30-2.50), mechanical ventilation on day 1 (aOR 2.65, 95% CI 1.88-3.71), and treatment with vasopressors on day 1 (aOR 3.23, 95% CI 2.19-4.75). Risk categories were low (no risk factor, 1.2%, 25/2137), moderate (singleton with no other risk factor: 1.8%, 68/3678), and high (≥1 risk factor among 5-minute Apgar <7, receipt of vasopressors or mechanical ventilation on day 1: 5.6%, 192/3408). Screening moderate- to high-risk infants identified 91% (260/285) of infants with severe neurologic injury and would require screening fewer infants (1647 infants per year) than screening all infants <33 weeks of gestation (2064 infants screened per year, 93% [265/285] of cases identified). CONCLUSIONS: Risk factor-based ultrasound scan of the head screening among infants born at 30-32 weeks of gestation could help optimize resources better than gestational age based screening.

Segmented Bayesian calibration approach for estimating age in forensic science.

Forensic age estimation is receiving growing attention from researchers in the last few years. Accurate estimates of age are needed both for identifying real age in individuals without any identity document and assessing it for human remains. The methods applied in such context are mostly based on radiological analysis of some anatomical districts and entail the use of a regression model. However, estimating chronological age by regression models leads to overestimated ages in younger subjects and underestimated ages in older ones. We introduced a full Bayesian calibration method combined with a segmented function for age estimation that relied on a Normal distribution as a density model to mitigate this bias. In this way, we were also able to model the decreasing growth rate in juveniles. We compared our new Bayesian-segmented model with other existing approaches. The proposed method helped producing more robust and precise forecasts of age than compared models while exhibited comparable accuracy in terms of forecasting measures. Our method seemed to overcome the estimation bias also when applied to a real data set of South-African juvenile subjects.

Real and predicted mortality under health spending constraints in Italy: a time trend analysis through artificial neural networks.

BACKGROUND: After 2008 global economic crisis, Italian governments progressively reduced public healthcare financing. Describing the time trend of health outcomes and health expenditure may be helpful for policy makers during the resources' allocation decision making process. The aim of this paper is to analyze the trend of mortality and health spending in Italy and to investigate their correlation in consideration of the funding constraints experienced by the Italian national health system (SSN). METHODS: We conducted a 20-year time-series study. Secondary data has been extracted from a national, institution based and publicly accessible retrospective database periodically released by the Italian Institute of Statistics. Age standardized all-cause mortality rate (MR) and health spending (Directly Provided Services - DPS, Agreed-Upon Services - TAUS, and private expenditure) were reviewed. Time trend analysis (1995-2014) through OLS and Multilayer Feed-forward Neural Networks (MFNN) models to forecast mortality and spending trend was performed. The association between healthcare expenditure and MR was analyzed through a fixed effect regression model. We then repeated MFNN time trend forecasting analyses on mortality by adding the spending item resulted significantly related with MR in the fixed effect analyses. RESULTS: DPS and TAUS decreased since 2011. There was a mismatch in mortality rates between real and predicted values. DPS resulted significantly associated to mortality (p < 0.05). In repeated mortality forecasting analysis, predicted MR was found to be lower when considering the pre-constraints health spending trend. CONCLUSIONS: Between 2011 and 2014, Italian public health spending items showed a reduction when compared to prior years. Spending on services directly provided free of charge appears to be the financial driving force of the Italian public health system. The overall mortality was found to be higher than the predicted trend and this scenario may be partially attributable to the healthcare funding constraints experienced by the SSN.

Transferring Health Big Data within the European Legal Framework: What Role for National Healthcare Services?

The main objective of this article is to describe the legal principles governing the selection by European public authorities, such as National Health Services (NHS) of third parties, when entering into agreements for the transfer of health data. According to Directive 2003/98/EC, and in light of the provisions of the Treaties of the European Union, the choice as to how a public authority makes its data available to third parties needs to be transparent, non-discriminatory and may not in any case benefit a specific company at the expense of others. For this reason, we maintain that a hypothetical agreement by which a public authority grants exclusive access to a large amount of health data to a private company selected with non-transparent criteria appears highly questionable. We advocate that the NHS should adopt more appropriate data policies aimed at promoting the sustainability of the NHS, following the legal framework analysed in this article.

Analyzing the 20-year declining trend of hospital length-of-stay in European countries with different healthcare systems and reimbursement models.

The study aims to investigate the last 20-year (2000-2019) of hospital length of stay (LOS) trends and their association with different healthcare systems (HS) among 25 European countries. A panel dataset was created using secondary data from Eurostat and Global Burden of Disease study databases, with dependent and control variables aggregated at the national level over a period of 20 years. A time trend analysis was conducted using a weighted least squares model for panel data to investigate the association between LOS, HS models [National Health Service (NHS), National Health Insurance, Social Health Insurance (SHI), and Etatist Social Health Insurance], healthcare reimbursement schemes [Prospective Global Budget (PGB), Diagnosis Related Groups (DRG), and Procedure Service Payment (PSP)], and control variables. The study showed a reduction of average LOS from 9.20 days in 2000 to 7.24 in 2019. SHI was associated with a lower LOS compared to NHS (b = - 0.6327, p < 0.05). Both DRG (b = 1.2399, p < 0.05) and PSP (b = 1.1677, p < 0.05) reimbursement models were positively associated with LOS compared to PGB. Our results confirmed the downward trend of LOS in the last 20 years, its multifactorial nature, and the influence of the SHI model of HS. This could be due to the financial incentives present in fee-for-service payment models and the role of competition in creating a market for healthcare services. These results offer insight into the factors influencing healthcare utilization and can inform the design of more effective, efficient, and sustainable HS.

Combining Bayesian Calibration and Copula Models for Age Estimation.

Accurately estimating and predicting chronological age from some anthropometric characteristics of an individual without an identity document can be crucial in the context of a growing number of forced migrants. In the related literature, the prediction of chronological age mostly relies upon the use of a single predictor, which is usually represented by a dental/skeletal maturity index, or multiple independent ordinal predictor (stage of maturation). This paper is the first attempt to combine a robust method to predict chronological age, such as Bayesian calibration, and the use of multiple continuous indices as predictors. The combination of these two aspects becomes possible due to the implementation of a complex statistical tool as the copula. Comparing the forecasts from our copula-based method with predictions from an independent model and two single predictor models, we showed that the accuracy increased.

A step-by-step researcher's guide to the use of an AI-based transformer in epidemiology: an exploratory analysis of ChatGPT using the STROBE checklist for observational studies.

Objective: This study aims at investigating how AI-based transformers can support researchers in designing and conducting an epidemiological study. To accomplish this, we used ChatGPT to reformulate the STROBE recommendations into a list of questions to be answered by the transformer itself. We then qualitatively evaluated the coherence and relevance of the transformer's outputs. Study design: Descriptive study. Methods: We first chose a study to be used as a basis for the simulation. We then used ChatGPT to transform each STROBE checklist's item into specific prompts. Each answer to the respective prompt was evaluated by independent researchers in terms of coherence and relevance. Results: The mean scores assigned to each prompt were heterogeneous. On average, for the coherence domain, the overall mean score was 3.6 out of 5.0, and for relevance it was 3.3 out of 5.0. The lowest scores were assigned to items belonging to the Methods section of the checklist. Conclusions: ChatGPT can be considered as a valuable support for researchers in conducting an epidemiological study, following internationally recognized guidelines and standards. It is crucial for the users to have knowledge on the subject and a critical mindset when evaluating the outputs. The potential benefits of AI in scientific research and publishing are undeniable, but it is crucial to address the risks, and the ethical and legal consequences associated with its use.

Optimizing timing and frequency of head ultrasound screening for severe brain injury among preterm infants born <32 weeks' gestation.

OBJECTIVE: To develop a head ultrasound (HUS) screening protocol for infants born <32 weeks gestational age (GA) that accurately identifies severe brain injury (SBI) while minimizing resource use. STUDY DESIGN: Retrospective cohort study of infants born <32 weeks GA, admitted to a level 3 neonatal intensive care unit between 2011 and 2017. Timing and results of each HUS were reviewed. SBI was defined as intraventricular hemorrhage grade ≥3 and/or periventricular leukomalacia. Logistic regression models were used to identify risk factors and evaluate the predictive value of HUS at different time points during hospitalization. RESULTS: Of 651 included infants, 71 (11%) developed SBI. Risk factors for SBI were GA at birth <29 weeks (adjusted odds ratio (aOR) 2.87, 95% confidence interval (CI) 1.50-5.48), vasopressors on admission (aOR 3.08, 95%CI 1.38-6.88) and mechanical ventilation on admission (aOR 2.50, 95%CI 1.33-4.68). Infants were classified into three risk groups based on these risk factors, and combinations of 1-5 HUS time points were evaluated to determine the optimal number and timing of HUS for each group. The optimal number of screening HUS ranged from 1 for low-risk to 2 for high-risk infants. Adopting a screening protocol using the number and timing of HUS optimized by risk group could reduce the total number of HUS performed by 40% and the median number of HUS per infant from 3 (IQR 2-4) to 2 (IQR 1-3) (p < .01). CONCLUSIONS: Implementation of a risk factor-based HUS screening protocol can help reduce resource use while maintaining high sensitivity for detecting SBI.

Association between Economic Growth, Mortality, and Healthcare Spending in 31 High-Income Countries.

This study aims to investigate the association between gross domestic product (GDP), mortality rate (MR) and current healthcare expenditure (CHE) in 31 high-income countries. We used panel data from 2000 to 2017 collected from WHO and OECD databases. The association between CHE, GDP and MR was investigated through a random-effects model. To control for reverse causality, we adopted a test of Granger causality. The model shows that the MR has a statistically significant and negative effect on CHE and that an increase in GDP is associated with an increase of CHE (p < 0.001). The Granger causality analysis shows that all the variables exhibit a bidirectional causality. We found a two-way relationship between GDP and CHE. Our analysis highlights the economic multiplier effect of CHE. In the debate on the optimal allocation of resources, this evidence should be taken into due consideration.

Variation of plasma citrulline as a predictive factor for weaning off long-term parenteral nutrition in children with neonatal short bowel syndrome.

BACKGROUND & AIMS: Long-term parenteral nutrition (PN) is the mainstay of the therapeutic strategy in intestinal failure (IF) due to neonatal short bowel syndrome (SBS). Our aim was to identify prognostic factors for PN weaning and to assess if measuring plasma citrulline concentrations over time could account for the intestinal adaptation in progress. METHODS: This retrospective study included children with neonatal SBS with surgical measurement of the residual bowel length and repeated plasma citrulline assessments during a 4-year follow-up. The degree of IF was assessed by the PN dependency index (PN caloric intake/Resting energy expenditure). The analysis was carried out according to SBS anatomical groups: end-jejunostomy (type 1), jejuno-colic (type 2) and jejuno-ileal anastomosis (type 3). RESULTS: Fifty-five patients (8 type 1, 27 type 2, 20 type 3) were included. None of the patients with SBS type 1, 11 (41%) with type 2 and 11 (55%) with type 3 were weaned off during the follow-up period. Plasma citrulline levels significantly increased with time in patients who were finally weaned off PN; conversely, the levels did not consistently increase in patients who were still on PN at the end of the study period. There was an inverse relationship between plasma citrulline levels and the PN dependency index. The increasing citrulline levels had a positive effect on the probability of weaning, 2.7 times higher for each point increase in citrulline. No significant effect of age and residual bowel length at baseline was found. CONCLUSION: The increased plasma citrulline level over time in addition to the SBS anatomical type is a reliable marker for subsequent PN weaning. The prediction of PN weaning assessed solely by the residual bowel length or a single measurement of citrulline is insufficient and should also take into account the anatomical type of SBS and repeated measurements of plasma citrulline levels.

Rivaroxaban for extended antithrombotic prophylaxis after laparoscopic surgery for colorectal cancer. Design of the PRO-LAPS II STUDY.

BACKGROUND: The clinical benefit of extending prophylaxis for venous thromboembolism (VTE) beyond hospital discharge after laparoscopic surgery for cancer is undefined. Extended prophylaxis with rivaroxaban is effective in reducing post-operative VTE after major orthopedic surgery without safety concern. METHODS: PROLAPS II is an investigator-initiated, randomized, double-blind study aimed at assessing the efficacy and safety of extended antithrombotic prophylaxis with rivaroxaban compared with placebo after laparoscopic surgery for colorectal cancer in patients who had received antithrombotic prophylaxis with low molecular-weight heparin for 7 ± 2 days (NCT03055026). Patients are randomized to receive rivaroxaban (10 mg once daily) or placebo for 3 weeks (up to day 28 ± 2 from surgery). The primary study outcome is a composite of symptomatic objectively confirmed VTE, asymptomatic ultrasonography-detected DVT or VTE-related death at 28 ± 2 days from laparoscopic surgery. The primary safety outcome is major bleeding defined according to the International Society of Thrombosis and Haemostasis. Symptomatic objectively confirmed VTE, asymptomatic ultrasonography-detected DVT, major bleeding or death by day 28 ± 2 and by day 90 from surgery are secondary outcomes. Assuming an 8% event rate with placebo and 60% reduction in the primary study outcome with rivaroxaban, 323 patients per group are necessary to show a statistically significant difference between the study groups. DISCUSSION: The PROLAPS II is the first study with an oral anti-Xa agent in cancer surgery. The study has the potential to improve clinical practice by answering the question on the clinical benefit of extending prophylaxis after laparoscopic surgery for colorectal cancer.

Mirror Neurons in the Macaque Monkey: A Critical Review.

In a number of experimental studies carried out in Italy in the early 1990s, it was observed that a set of neurons from the macaque monkey premotor cortex is activated both when a specific action is performed and when the same action is simply observed. These neurons were given the evocative name of "mirror neurons". In this article, the conclusions from these studies and subsequent elaborations are reviewed in order to examine their soundness in theoretical terms.

Treatment of splenic flexure colon cancer: a comparison of three different surgical procedures: Experience of a high volume cancer center.

Extended right or left hemicolectomy are the most common surgical treatments for splenic flexure colon cancer. Extended resection (including distal pancreasectomy and/or splenectomy), has been often indicated for the treatment for the splenic flexure cancer, because the lymphatic drainage at this site is poorly defined and assumed as heterogeneous. Between January 2006 and May 2016, 103 patients with splenic flexure colon cancer were enrolled in the study. We evaluated the clinicopathological findings and outcomes of all patients and associated them to the different surgical treatment. Out of 103 selected cases an extended right hemicolectomy was performed in 22 (21.4%) patients, an extended left hemicolectomy in 24 (23.3%) patients, a segmental resection of the splenic flexure in 57 (55.3%) patients; the combined resection of adjacent organs showing tumor adherence was carried out in 11 (10.7%) patients. The tumor infiltrated near organs (T4) in 5 patients. No significant differences in complications were found among the three groups. In all groups no differences were found in the total number of harvested lymphnodes. After a median follow-up of 42 months, 30 recurrences and 19 deaths occurred (12 for tumor progression). There was no difference in overall and progression free survival among the three different surgical treatments. According to our results, the partial resection of splenic flexure was not associated with a worse prognosis and it was leading for a satisfactory oncological outcome. It is our opinion that the extended surgery is seldomly indicated to cure splenic flexure cancer.

Health Expenditure and All-Cause Mortality in the 'Galaxy' of Italian Regional Healthcare Systems: A 15-Year Panel Data Analysis.

BACKGROUND: The sustainability of healthcare systems is a topic of major interest. During periods of economic instability, policy makers typically reallocate resources and execute linear cuts in different areas of public spending, including healthcare. OBJECTIVES: The aim of this paper was to examine whether and how per capita public healthcare expenditure (PHE) in the Italian regions was related to the all-cause mortality rate (MR) between 1999 and 2013 and to determine which expenditure item most affected mortality in the short and very short term. METHODS: We conducted a pooled cross-sectional time series study. Secondary data were extracted from 'Health for All', a database released periodically by the Italian National Institute of Statistics. PHE is subdivided into directly provided services (DPS), pharmaceutical care, general practitioner care, specialist medical care, privately delivered hospital care, other privately delivered medical services, and psychiatric support and rehabilitation. We used a fixed-effects regression to assess the effects of PHE items on the MR after controlling for a number of socioeconomic and supply variables. RESULTS: Higher spending on DPS was associated with a lower MR. Other expenditure variables were not significantly associated with the MR. CONCLUSIONS: The results highlight the importance of medical services and goods provided directly by public services (i.e. hospital-based general and specialized wards and offices, emergency departments, etc.). DPS represents the driving force of the system and should be considered a determinant of the health of the Italian population. Our results suggest that the context and financing methods of a healthcare system should be carefully analysed before linear cuts are made or resources are reallocated.