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Summary: Single-cell spatial analysis of proteins is rapidly becoming increasingly important in revealing biological insights. Here, we present a protocol for automated high-plex multi-slide immunofluorescence staining and imaging of human head and neck cancer formalin-fixed paraffin-embedded (FFPE) sections using PhenoCycler-Fusion 2.0 technology. We describe steps for preparing human head and neck cancer FFPE tissues, staining with a panel of immunophenotyping markers, and Flow Cell assembly. We then detail procedures for setting up for a PhenoCycler-Fusion run, post-run Flow Cell removal, and downstream analyses.For complete details on the use and execution of this protocol, please refer to Jhaveri et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Morocco is currently facing significant challenges due to the ever–changing climate, with its critical water sources crucial for agriculture, economy, and daily life being greatly affected. In order to thoroughly understand the impact of climate change on the Ghis–Nekor watershed, an in–depth study spanning 38 years (1978–2016) was conducted. This involved examining meteorological data from three stations and utilizing advanced indices such as SPI, RDI, and DI. The findings of this study reveal prominent shifts in precipitation patterns, indicating a vulnerability in the region. While there was a general increase in annual rainfall during the specified time period, a sharp decline was observed post–2008. Further analysis of drought confirmed the presence of persistent dry spells and recurring episodes, highlighting the urgent need for effective water management strategies. These crucial findings must be considered by decision–makers for successful climate adaptation, emphasizing the crucial role played by this study in mitigating the effects of climate change.

Abstract Introduction Adverse events (AEs) that occur in hospitals remain a challenge worldwide, and especially in intensive care units (ICUs) where they are more likely to occur. Monitoring of AEs can provide insight into the status and advances of patient safety. This study aimed to examine the AEs reported during the 20 months after the implementation of the AE reporting system. Methods We conducted a retrospective analysis of a voluntary ICU AE reporting system. Incidents were reported by the staff from ten ICUs in the Sahloul University Hospital (Tunisia) between February 2020 and September 2021. Results A total of 265 reports were received, of which 61.9% were deemed preventable. The most frequently reported event was healthcare-associated infection (30.2%, n = 80), followed by pressure ulcers (18.5%, n = 49). At the time of reporting, 25 patients (9.4%) had died as a result of an AE and in 51.3% of cases, the event had resulted in an increased length of stay. Provider-related factors contributed to 64.2% of the events, whilst patient-related factors contributed to 53.6% of the events. As for criticality, 34.3% of the events (n = 91) were unacceptable (c3) and 36.3% of the events (n = 96) were ‘acceptable under control’ (c2). Conclusions The reporting system provided rich information on the characteristics of reported AEs that occur in ICUs and their consequences and may be therefore useful for designing effective and evidence-based interventions to reduce the occurrence of AEs.

Abstract Cell annotation is a crucial methodological component to interpreting single cell and spatial omics data. These approaches were developed for single cell analysis but are often biased, manually curated and yet unproven in spatial omics. Here we apply a stemness model for assessing oncogenic states to single cell and spatial omic cancer datasets. This one-class logistic regression machine learning algorithm is used to extract transcriptomic features from non-transformed stem cells to identify dedifferentiated cell states in tumors. We found this method identifies single cell states in metastatic tumor cell populations without the requirement of cell annotation. This machine learning model identified stem-like cell populations not identified in single cell or spatial transcriptomic analysis using existing methods. For the first time, we demonstrate the application of a ML tool across five emerging spatial transcriptomic and proteomic technologies to identify oncogenic stem-like cell types in the tumor microenvironment.

Background. COVID-19 is the major cause of serious acute respiratory complications, leading to strong perturbation of cytokine release and high levels of interleukin-6. In this work, we assessed the efficacy and safety of Tocilizumab in Tunisian patients with severe COVID-19. Materials and Methods. We enrolled in this retrospective study 255 critically ill patients in the intensive care unit of the military hospital of Tunis. These patients were categorized into a standard group (n = 148) and a Tocilizumab group (n = 107). Medical records were gathered and analyzed between September, 2020 and September, 2021. Results. No significant difference was detected in clinical and biological parameters at admission between the two studied groups. Interestingly, we revealed here a significant improvement in respiratory parameters as well as biological findings such as hemoglobin (p=0.001), platelets (p=0.005), CRP (p=0.0001), fibrinogen (p=0.0001), and creatinine (p=0.002). Regarding the Simplified Acute Physiology Score II and 28-day mortality, we showed a significant decrease of these two parameters following 48 hr of drug administration (p=0.0001, respectively). No significant difference between the two groups regarding complications except the renal failure and the need for renal replacement therapy, which are higher in the standard group as compared to patients treated with Tocilizumab (p=0.02; p=0.0001, respectively). Conclusions. Treatment with TCZ seems to be safe and effective in reducing the mortality rate in severe COVID-19 patients.

Spring black stem and leaf spot, caused by Phoma medicaginis, is an issue in annual Medicago species. Therefore, in this study, we analyzed the response to P. medicaginis infection in a collection of 46 lines of three annual Medicago species (M. truncatula, M. ciliaris, and M. polymorpha) showing different geographic distribution in Tunisia. The reaction in the host to the disease is explained by the effects based on plant species, lines nested within species, treatment, the interaction of species × treatment, and the interaction of lines nested within species × treatment. Medicago ciliaris was the least affected for aerial growth under infection. Furthermore, the largest variation within species was found for M. truncatula under both conditions. Principal component analysis and hierarchical classification showed that M. ciliaris lines formed a separate group under control treatment and P. medicaginis infection and they are the most vigorous in growth. These results indicate that M. ciliaris is the least susceptible in response to P. medicaginis infection among the three Medicago species investigated here, which can be used as a good candidate in crop rotation to reduce disease pressure in the field and as a source of P. medicaginis resistance for the improvement of forage legumes.

Introduction: Healthcare activities are generally associated with the production of healthcare waste, a large part of which is assimilated to household waste (packaging, kitchen waste, green waste, etc.) and another category of waste which may have a risk to health and the environment given its nature and typology. This category of waste at risk includes waste at risk of infection such as stinging, sharp waste (needles, blade, scalpel, etc.), and waste at chemical risk. Poor management of hospital waste is a problem in most countries and especially in developing countries. We aimed to determine the health and environmental impacts of the poor management of healthcare waste. Methods: We carried out a systematic review of the French and English literature on the scientific research sites Medline/PubMed and Embase. This research was carried out over 3 months (April–June 2020). The search strategy was used by combining keywords and Boolean operators: Health, Health impact assessment, Hospitals, Medical waste, Waste disposal facilities, Environment, Environment/Epidemiology, Hospital waste, impact, workplace, Environment hazards, Healthcare works, Waste management. Results: It has been clear that the current management of healthcare waste is not capable of adequately preserving human health and environmental contamination from infection. The surveys analyzed showed that if incineration is properly treated, it would be an appropriate treatment method to deal with healthcare waste. However, exposure to pollutants produced by the incineration is still a public health problem. If incineration is seen as a practical solution for dealing with healthcare waste, low-temperature incinerators should be banned and replaced by modern incinerators equipped with air pollution control units. These problems are typical for any developing country which does not have the means to purchase incinerators which are more protective for the environment and equipped with the latest technologies. Conclusions: Thus, autoclaving and microwaves are considered better alternatives for treating healthcare waste. However, these methods are generally not adequate for the disposal of pathological, radioactive, laboratory, and chemotherapy wastes. Therefore, the specific management of healthcare waste is a major concern due to the potentially high risks for human health and the environment.

In the context of the challenges imposed by climate change, which exerts constant pressure on water security in Morocco, the Moulouya River, one of the main sources of surface water, frequently undergoes spatiotemporal variations in its quality. This situation is particularly worrying in the upstream part of the river, where the old Zaida mine is located, which is now inactive and leaves significant residues in the form of slag heaps in direct contact with the waters of the Moulouya River. To assess the impact of this abandoned mining waste, eight (08) water sampling stations (S1 to S8) were chosen to analyse the concentrations of two (02) essential metals, namely, lead and zinc. The results revealed lead concentrations of approximately 0.13 mg/L on average during the dry period and 0.09 mg/L during the humid period. The average zinc concentration was 0.29 mg/L during the dry season and 0.32 mg/L during the humid season. The maximum concentrations were recorded at the stations closest to the aforementioned mine, in particular, S1, S2 and S3. Other maximum concentrations were observed at station S6. These results highlighted the significant impact of mining slag heaps exposed to the open sky on the physico-chemical and metallic quality of Moulouya waters. This finding highlights the urgency of continued monitoring of this vital water resource to protect it against potential risks of contamination.

Acute necrotic pancreatitis is an emergency of evolution and is often unpredictable because of the potentially life-threatening complications it can cause. We report a unique case of a 56-year-old woman hospitalized for acute necrotic pancreatitis. The evolution of the latter was characterized by the occurrence of two very rare complications, of which the clinical presentations were atypical. The first complication was a gastroduodenal pseudoaneurysm compressing the main biliary tract and causing obstructive jaundice, which evolved well following percutaneous embolization. The second complication was a giant 20 cm pancreatic pseudocyst revealed by obstructive jaundice secondary to biliary compression, which progressed well following surgical treatment.