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Abstract Background The clinical potential of Raman spectroscopy is well established but has yet to become established in routine oncology workflows. One barrier slowing clinical adoption is a lack of evidence demonstrating that data taken on one spectrometer transfers across to data taken on another spectrometer to provide consistent diagnoses. Methods We investigated multi-centre transferability using human oesophageal tissue. Raman spectra were taken across three different centres with different spectrometers of the same make and model. By using a common protocol, we aimed to minimise the difference in machine learning performance between centres. Results 61 oesophageal samples from 51 patients were interrogated by Raman spectroscopy at each centre and classified into one of five pathologies. The overall accuracy and log-loss did not significantly vary when a model trained upon data from any one centre was applied to data taken at the other centres. Computational methods to correct for the data during pre-processing were not needed. Conclusion We have found that when using the same make and model of spectrometer, together with a common protocol, across different centres it is possible to achieve system transferability without the need for additional computational instrument correction.

In situ cryo-electron tomography (cryo-ET) is the most current, state-of-the-art technique to study cell machinery in its hydrated near-native state. The method provides ultrastructural details at sub-nanometer resolution for many components within the cellular context. Making use of recent advances in sample preparation techniques and combining this method with correlative light and electron microscopy (CLEM) approaches have enabled targeted molecular visualization. Nevertheless, the implementation has also added to the complexity of the workflow and introduced new obstacles in the way of streamlining and achieving high throughput, sample yield, and sample quality. Here, we report a detailed protocol by combining multiple newly available technologies to establish an integrated, high-throughput, optimized, and streamlined cryo-CLEM workflow for improved sample yield.Key features• PRIMO micropatterning allows precise cell positioning and maximum number of cell targets amenable to thinning with cryo focused-ion-beam–scanning electron microscopy.• CERES ice shield ensures that the lamellae remain free of ice contamination during the batch milling process.• METEOR in-chamber fluorescence microscope facilitates the targeted cryo focused-ion-beam (cryo FIB) milling of these targets.• Combining the three technologies into one cryo-CLEM workflow maximizes sample yield, throughput, and efficiency.Graphical overview

Extracellular vesicles (EVs) are a heterogeneous population of lipid bilayer membrane-bound vesicles which encapsulate bioactive molecules, such as nucleic acids, proteins, and lipids. They mediate intercellular communication through transporting internally packaged molecules, making them attractive therapeutics carriers. Over the last decades, a significant amount of research has implied the potential of EVs servings as drug delivery vehicles for nuclear acids, proteins, and small molecular drugs. However, several challenges remain unresolved before the clinical application of EV-based therapeutics, including lack of specificity, stability, biodistribution, storage, large-scale manufacturing, and the comprehensive analysis of EV composition. Technical development is essential to overcome these issues and enhance the pre-clinical therapeutic effects. In this review, we summarize the current advancements in EV engineering which demonstrate their therapeutic potential.

Abstract Background Most of the existing research on supervised consumption services (SCS) is focused on injection drug use. Less is known about the applicability of SCS for people who consume drugs orally, intranasally, or through inhalation. This is problematic because people who use drugs through modes other than injection are also at risk of overdose death and other harm, and experience barriers accessing health and social services. We aimed to describe existing SCS models that accommodate these alternate routes of drug consumption, and synthesize available information on characteristics of program participants. Methods We conducted a systematic scoping review of 9 peer-reviewed and 13 grey literature databases on SCS that incorporate non-injection routes of consumption. We screened 22,882 titles, and excluded 22,843 (99.8%) articles. We ultimately included 39 (0.2%) full-text articles; 28 (72%) of these articles explicitly identified SCS that permit alternate routes of consumption and 21 (54%) discussed characteristics of participants who consume drugs through non-injection routes. Data on study characteristics, terms and definitions, and site and program participant characteristics were extracted and double-coded. Extracted data were analyzed using descriptive statistics and narrative synthesis. Results Included articles describe 48 SCS that permit non-injection routes of consumption, most of which were located in Germany. The majority of these SCS were legally sanctioned and had models of care that were largely comparable to supervised injection services. Notable differences included physical infrastructure such as ventilated rooms or outdoor areas to accommodate inhalation, and shorter time limits on non-injection drug consumption episodes. Program participants engaging in non-injection forms of consumption were typically men over the age of 30 and structurally vulnerable (e.g., experiencing homelessness or unstable housing). Conclusions Extant academic and grey literature indicates that site characteristics and demographics of program participants of SCS that permit non-injection routes of consumption largely reflect those of supervised injection services. Further research on the range of existing SCS that incorporate non-injection routes of consumption is needed to ensure high quality service provision, and improved health outcomes for people who consume drugs via oral, intranasal, and inhalation routes.

Introduction: Perinatal asphyxia is one of the main causes of morbidity and mortality in newborns. It generates high costs, both social and economic, and presents modifiable risk factors.Objective: To determine the biological and psychosocial factors and risk behaviors associated with the development of perinatal asphyxia (Sarnat II-III) in newborns from low socioeconomic status in a tier III university hospital in the city of Cali, Colombia.Materials and Methods: With a case and control design, 216 patients were studied (54 cases/162 controls) (1 case/3 matched controls). The cases were defined as newborns with modified or severe perinatal asphyxia (Sarnat II-III) between 2012 and 2014, with gestational age ≥ 36 weeks, with neurological signs not attributable to other causes, multiorgan compromise, advanced reanimation, and presence of a sentinel event. For the analysis, conditional logistic regression models were developed to evaluate association (OR), considering that the cases and controls had been paired by the birth and gestational age variables.Results: The final model showed that, from the group of biological variables, meconium amniotic fluid was identified as a risk factor (OR 15.28, 95%CI 2.78–83.94). Induction of labor lowered the risk of perinatal asphyxia by 97% (OR 0.03, 95%CI 0.01–0.21), and monitoring of fetal heart rate was associated with lower odds by 99% (OR 0.01, 95%CI 0.00–0.31) of developing perinatal asphyxia in the newborn. Regarding social variables, the lack of social support was identified as a risk factor for the development of perinatal asphyxia (OR 6.44, 95%CI 1.16–35.66); in contrast, secondary education lowered the odds of developing perinatal asphyxia by 85% when compared with pregnant women who only had primary school education (OR 0.15, 95%CI 0.03–0.77).Conclusion: Assessment of biological and psychosocial factors and social support is important in pregnant women to determine the risk of developing perinatal asphyxia in a low-income population.

Background: Extracellular vesicles (EVs) are attracting interest as a new class of drug delivery vehicles due to their intrinsic nature of biomolecular transport in the body. We previously demonstrated that EV surface modification with tissue-specific molecules accomplished targeted EV-mediated DNA delivery. Methods: Here, we describe reliable methods for (i) generating EGFR tumor-targeting EVs via the display of high-affinity monobodies and (ii) in vitro measurement of EV binding using fluorescence and bioluminescence labeling. Monobodies are a well-suited class of small (10 kDa) non-antibody scaffolds derived from the human fibronectin type III (FN3) domain. Results: The recombinant protein consists of the EGFR-targeting monobody fused to the EV-binding domain of lactadherin (C1C2), enabling the monobody displayed on the surface of the EVs. In addition, the use of bioluminescence or fluorescence molecules on the EV surface allows for the assessment of EV binding to the target cells. Conclusions: In this paper, we describe methods of EV engineering to generate targeted delivery vehicles using monobodies that will have diverse applications to furnish future EV therapeutic development, including qualitative and quantitative in vitro evaluation for their binding capacity.