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Candida auris is an emerging fungal pathogen notable for its resistance to multiple antifungals and ability to survive in various environments. Understanding the interactions between C. auris and environmental protozoa, such as Acanthamoeba castellanii, could provide insights into fungal adaptability and pathogenicity. Two C. auris isolates (MMC1 and MMC2) were co-cultured with A. castellanii to examine interaction dynamics, survival, stress responses, growth, virulence, biofilm formation, and antifungal susceptibility. The association of C. auris-A. castellanii varied with a multiplicity of infection (MOI), with MMC2 exhibiting higher association rates at increased MOI than MMC1. Both isolates survived distinctly within A. castellanii , as the MMC1 showed an initial decline and subsequent increase in viability, while MMC2 maintained higher viability for up to 24 h, decreasing afterward. Both isolates exhibited accelerated growth when recovered from A. castellanii . The MMC2 isolate displayed increased resistance to oxidative, osmotic, and thermal stresses upon interaction with A. castellanii , whereas MMC1 showed limited changes. Exposure to A. castellanii also influenced the expression of virulence factors differently, with MMC1 increasing phospholipase and peptidase, while MMC2 upregulated phytase, esterase, hemolysin, and siderophores. Upon contact with A. castellanii , MMC2 enhanced biofilm formation, unlike MMC1. Both isolates increased ergosterol upon interactions, enhancing susceptibility to amphotericin B. However, both isolates were more tolerant to itraconazole and caspofungin, particularly MMC2, which showed differential expression of ergosterol biosynthesis enzymes and increased cell wall polysaccharides. This study reveals that interactions with A. castellanii modulate C. auris physiology and virulence, contributing to its environmental adaptability and resistance to antifungals., Importance: Candida auris has emerged as a critical public health concern due to its resistance to multiple antifungal drugs and ability to survive on surfaces under harsh conditions, mainly due to biofilm formation. The precise origin of this emerging pathogen still awaits elucidation, but interactions with environmental protozoa may have helped C. auris to develop such virulence and resistance traits. In this work, we precisely characterize the interactions of C. auris with the free-living amoeba Acanthamoeba castellanii and how these protozoa may alter the fungal behavior in terms of virulence, thermotolerance, biofilm formation capacity, and drug resistance. It may be essential to understand the various interactions C. auris could perform in the environment, directly impacting the outcome of human infections under the One Health approach.

Background and Purpose: Although there is extensive evidence about the safety of monoclonal antibodies against calcitonin gene-related peptide (anti-CGRP mAbs) in combination with traditional drugs, scarce data are available on the safety of their combination with other mAbs. This study aimed to evaluate the 6-month effectiveness and tolerability of anti-CGRP mAbs in combination with other mAbs for different diseases., Methods: Patients included in the Italian Headache Registry and treated concomitantly with an anti-CGRP mAb and another mAb were included. Effectiveness outcomes for migraine included reduction from baseline of monthly headache days (MHDs), Migraine Disability Assessment (MIDAS) score, Headache Impact Test-6 (HIT-6) scores, and Patients' Global Impression of Change (PGIC) scale. Adverse events (AEs) were recorded., Results: Thirty-eight patients were included. In 27 patients (71.1%), the anti-CGRP mAb was added to a previously ongoing mAb. Nine patients (23.7%) discontinued one of the two mAbs before the end of treatment (seven discontinued the anti-CGRP mAb and two the other mAb). One patient discontinued for AEs. Anti-CGRP mAbs were discontinued due to ineffectiveness (n = 5, 55.5%) and one each (11.1%) for clinical remission and lost to follow-up. MHDs significantly decreased from baseline to 3 months (p < 0.0001) and 6 months (p < 0.001), as did the MIDAS and the HIT-6 scores at 3 and 6 months (p < 0.001). For anti-CGRP mAbs, 27.4% of patients reported PGIC ≥ 5 at 3 months and 48.3% at 6 months. Mild AEs associated with introduction of a second mAb were detected in six patients (15.8%)., Conclusions: In this real-world study, anti-CGRP mAbs showed safety and effectiveness when administered concomitantly with other mAbs., (© 2024 The Author(s). European Journal of Neurology published by John Wiley & Sons Ltd on behalf of European Academy of Neurology.)

We conducted a comprehensive comparative analysis of extracellular vesicles (EVs) from two Acanthamoeba castellanii strains, Neff (environmental) and T4 (clinical). Morphological analysis via transmission electron microscopy revealed slightly larger Neff EVs (average = 194.5 nm) compared to more polydisperse T4 EVs (average = 168.4 nm). Nanoparticle tracking analysis (NTA) and dynamic light scattering validated these differences. Proteomic analysis of the EVs identified 1,352 proteins, with 1,107 common, 161 exclusive in Neff, and 84 exclusively in T4 EVs. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) mapping revealed distinct molecular functions and biological processes and notably, the T4 EVs enrichment in serine proteases, aligned with its pathogenicity. Lipidomic analysis revealed a prevalence of unsaturated lipid species in Neff EVs, particularly triacylglycerols, phosphatidylethanolamines (PEs), and phosphatidylserine, while T4 EVs were enriched in diacylglycerols and diacylglyceryl trimethylhomoserine, phosphatidylcholine and less unsaturated PEs, suggesting differences in lipid metabolism and membrane permeability. Metabolomic analysis indicated Neff EVs enrichment in glycerolipid metabolism, glycolysis, and nucleotide synthesis, while T4 EVs, methionine metabolism. Furthermore, RNA-seq of EVs revealed differential transcript between the strains, with Neff EVs enriched in transcripts related to gluconeogenesis and translation, suggesting gene regulation and metabolic shift, while in the T4 EVs transcripts were associated with signal transduction and protein kinase activity, indicating rapid responses to environmental changes. In this novel study, data integration highlighted the differences in enzyme profiles, metabolic processes, and potential origins of EVs in the two strains shedding light on the diversity and complexity of A. castellanii EVs and having implications for understanding host-pathogen interactions and developing targeted interventions for Acanthamoeba -related diseases.IMPORTANCEA comprehensive and fully comparative analysis of extracellular vesicles (EVs) from two Acanthamoeba castellanii strains of distinct virulence, a Neff (environmental) and T4 (clinical), revealed striking differences in their morphology and protein, lipid, metabolites, and transcripts levels. Data integration highlighted the differences in enzyme profiles, metabolic processes, and potential distinct origin of EVs from both strains, shedding light on the diversity and complexity of A. castellanii EVs, with direct implications for understanding host-pathogen interactions, disease mechanisms, and developing new therapies for the clinical intervention of Acanthamoeba -related diseases., Competing Interests: The authors declare no conflict of interest.

The importance of humoral immunity to fungal infections remains to be elucidated. In cryptococcosis, patients that fail to generate antibodies against antigens of the fungus Cryptococcus neoformans are more susceptible to the disease, demonstrating the importance of these molecules to the antifungal immune response. Historically, antibodies against C. neoformans have been applied in diagnosis, therapeutics, and as important research tools to elucidate fungal biology. Throughout the process of generating monoclonal antibodies (mAbs) from a single B-cell clone and targeting a single epitope, several immunization steps might be required for the detection of responsive antibodies to the antigen of interest in the serum. This complex mixture of antibodies comprises the polyclonal antibodies. To obtain mAbs, B-lymphocytes are harvested (from spleen or peripheral blood) and fused with tumor myeloma cells, to generate hybridomas that are individually cloned and specifically screened for mAb production. In this chapter, we describe all the necessary steps, from the immunization to polyclonal antibody harvesting, hybridoma generation, and mAb production and purification. Additionally, we discuss new cutting-edge approaches for generating interspecies mAbs, such as humanized mAbs, or for similar species in distinct host backgrounds., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Introduction: EASIER is a multicenter, observational, cross-sectional study investigating the consumption of healthcare resources, including healthcare professional (HCP) active working time, the costs associated with the current natalizumab intravenous (IV) administration, and the potential impact of the adoption of subcutaneous (SC) route., Methods: The EASIER study has three parts: (1) time and motion study to measure healthcare resources and working time needed for natalizumab IV administration using a digital data collection tool operated directly by HCPs; (2) HCP structured questionnaire-based estimation of the potential impact of natalizumab SC vs. IV administration; and (3) patient survey on the burden of natalizumab administration., Results: Nine Italian multiple sclerosis (MS) centers measured 404 IV natalizumab administration procedures and administered 26 HCP questionnaires and 297 patient questionnaires. Patients had a mean of 52 (range 1-176) previous IV administrations and spent a mean (median, IQR) of 152 (130, 94-184) minutes in the center per each IV procedure, with IV infusion covering 50% of the total. Including patient travel time, an average of 5 h was dedicated to each IV administration. Active working time by HCP amounted to 29 min per IV administration procedure, 70% of which by nursing staff. With adoption of the SC route, HCPs estimated a 50% reduction in patient procedure time and 55% lower HCP active working time. This translated into a 63% cost reduction for the MS center per natalizumab administration procedure., Conclusions: SC natalizumab administration will consistently reduce consumption of patient and HCP times per procedure and associated costs., (© 2023. The Author(s).)

Introduction: The progressive loss of skeletal muscle mass, strength, and function that frequently occurs as people get older is referred to as sarcopenia. Elderly musculoskeletal aging, sarcopenia, and obesity are all intimately connected. Our study's aim is to investigate the prevalence of sarcopenia in a real cohort of patients over 65 with musculoskeletal conditions referring to a Rehabilitation Unit. The secondary aim of our study is to investigate associations between sarcopenia and alterations in nutritional status and Body Mass Index (BMI). Finally, quality of life and global health has been investigated in our population., Materials and Methods: From January 2019 to January 2021, 247 patients over 65 years old with musculoskeletal concerns were enrolled and participated in an observational study. As outcome measures, the Mini Nutritional Assessment (MNA), the 12-Item Short Form Health Survey (SF-12), and the Cumulative Illness Rating Scale Severity Index (CIRS-SI) were used. Additionally, measurements of total skeletal muscle mass (SMM) and appendicular muscle mass (ASMM) using bioelectrical impedance analysis, as well as a hand grip strength test of the non-dominant hand were taken. The Mid Upper Arm Circumference (MUAC) and the Calf Circumference (CC) were measured and recorded as further indications of possible sarcopenia., Results: A percentage of 46.1% of subjects with overt sarcopenia was found and 10.1% showed a severe sarcopenia. Patients with severe sarcopenia showed significantly lower values of BMI and MNA. Additionally, sarcopenic patients showed significantly lower values in MNA when compared to non-sarcopenic patients. Considering SF-12, only the physical score revealed slight significant differences. In particular, patients affected by probable or severe sarcopenia presented a lower value than non-sarcopenic patients. Concerning MUAC and CC, severe sarcopenic patients showed significant lower values for both the body parts., Conclusion: Our study considers a cohort of real-life elderly subjects with musculoskeletal concerns and shows that these subjects are highly susceptible to sarcopenia. Therefore, rehabilitation for elderly patients with musculoskeletal concerns requires to be customized and multidisciplinary. Future research should further investigate these aspects in order to enable the early identification of sarcopenia and the formulation of customized rehabilitative programs. ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Maccarone, Coraci, Bernini, Sarandria, Valente, Frigo, Dionyssiotis and Masiero.)

Functional imaging experimental designs measuring fatigue, defined as a subjective lack of physical and/or mental energy characterizing a wide range of neurologic conditions, are still under development. Nineteen right-handed healthy subjects (9 M and 10 F, mean age 43.15 ± 8.34 years) were evaluated by means of functional magnetic resonance imaging (fMRI), asking them to perform explicit, first-person, mental imagery of fatigue-related multisensory sensations. Short sentences designed to assess the principal manifestations of fatigue from the Multidimensional Fatigue Symptom Inventory were presented. Participants were asked to imagine the corresponding sensations (Sensory Imagery, SI). As a control, they had to imagine the visual scenes (Visual Imagery, VI) described in short phrases. The SI task (vs. VI task) differentially activated three areas: (i) the precuneus, which is involved in first-person perspective taking; (ii) the left superior temporal sulcus, which is a multisensory integration area; and (iii) the left inferior frontal gyrus, known to be involved in mental imagery network. The SI fMRI task can be used to measure processing involved in mental imagery of fatigue-related multisensory sensations., (© 2022 The Authors. Human Brain Mapping published by Wiley Periodicals LLC.)

Background: Efficiency of care chain response and hospital reactivity were and are challenged for stroke acute care management during the pandemic period of coronavirus disease 2019 (COVID-19) in North-Eastern Italy (Veneto, Friuli-Venezia-Giulia, Trentino-Alto-Adige), counting 7,193,880 inhabitants (ISTAT), with consequences in acute treatment for patients with ischemic stroke., Methods: We conducted a retrospective data collection of patients admitted to stroke units eventually treated with thrombolysis and thrombectomy, ranging from January to May 2020 from the beginning to the end of the main first pandemic period of COVID-19 in Italy. The primary endpoint was the number of patients arriving to these stroke units, and secondary endpoints were the number of thrombolysis and/or thrombectomy. Chi-square analysis was used on all patients; furthermore, patients were divided into two cohorts (pre-lockdown and lockdown periods) and the Kruskal-Wallis test was used to test differences on admission and reperfusive therapies., Results: In total, 2536 patients were included in 22 centers. There was a significant decrease of admissions in April compared to January. Furthermore, we observed a significant decrease of thrombectomy during the lockdown period, while thrombolysis rate was unaffected in the same interval across all centers., Conclusions: Our study confirmed a decrease in admission rate of stroke patients in a large area of northern Italy during the lockdown period, especially during the first dramatic phase. Overall, there was no decrease in thrombolysis rate, confirming an effect of emergency care system for stroke patients. Instead, the significant decrease in thrombectomy rate during lockdown addresses some considerations of local and regional stroke networks during COVID-19 pandemic evolution., (© 2021. The Author(s).)

Although antibiotic-induced dysbiosis has been demonstrated to exacerbate intestinal inflammation, it has been suggested that antibiotic prophylaxis may be beneficial in certain clinical conditions such as acute pancreatitis (AP). However, whether broad-spectrum antibiotics, such as meropenem, influence the dissemination of multidrug-resistant (MDR) bacteria during severe AP has not been addressed. In the currently study, a mouse model of obstructive severe AP was employed to investigate the effects of pretreatment with meropenem on bacteria spreading and disease outcome. As expected, animals subjected to biliopancreatic duct obstruction developed severe AP. Surprisingly, pretreatment with meropenem accelerated the mortality of AP mice (survival median of 2 days) when compared to saline-pretreated AP mice (survival median of 7 days). Early mortality was associated with the translocation of MDR strains, mainly Enterococcus gallinarum into the blood stream. Induction of AP in mice with guts that were enriched with E. gallinarum recapitulated the increased mortality rate observed in the meropenem-pretreated AP mice. Furthermore, naïve mice challenged with a mouse or a clinical strain of E. gallinarum succumbed to infection through a mechanism involving toll-like receptor-2. These results confirm that broad-spectrum antibiotics may lead to indirect detrimental effects during inflammatory disease and reveal an intestinal pathobiont that is associated with the meropenem pretreatment during obstructive AP in mice.

Unlabelled: Increasing recent evidence favors paracetamol use for patent ductus arteriosus (PDA) closure in preterms. Our study aims were (1) to assess efficacy and safety of intravenous (i.v.) paracetamol for PDA closure in a 23-32-week preterm population, as "first-line" (when traditional ibuprofen treatment was contraindicated) or "rescue" treatment (after ibuprofen failed), and (2) to identify predictors of PDA closure. The cumulative efficacy of consecutive cycles of i.v. paracetamol on PDA closure was confirmed after both "first-line" and "rescue" treatment, the overall PDA closure rates being, respectively, 56.7 and 61.1 % (p = 0.7624) after two cycles and 63.3 and 77.8 % (p = 0.2959) after three cycles. No toxicity was apparent after either "first-line" or "rescue" i.v. paracetamol treatment. On multivariate analysis, gestational age (GA) emerged as an independent predictor of PDA closure in the "first-line" i.v. paracetamol treatment group, while clinical risk index for babies (CRIB) score (a patient risk index based on birth weight, GA at birth, sex, patient's temperature on admission, and maximum base excess in first 12 h of life) was an independent predictor of PDA closure failure in the "rescue" group., Conclusion: I.V. paracetamol proved effective in our study population. Randomized control trials (RCTs) are warranted to further investigate the efficacy and safety of i.v. paracetamol for PDA closure in preterms., What Is Known: • Oral paracetamol has been judged as effective as oral ibuprofen for PDA closure in the preterm. • To date, only a handful of non-randomized studies exist to support the effectiveness of i.v. paracetamol in PDA closure. What is New: • Our observations confirm the clinical efficacy of i.v. paracetamol for PDA closure in a very low birth weight (VLBW)/extremely low birth weight (ELBW) preterm population. • Gestational age and CRIB score emerge as independent predictors of PDA closure.

Cecropia species are widely used in traditional medicine by its anti-diabetic, anti-hypertensive and anti-inflammatory properties. In the present study, we investigated the neuroprotective and antioxidant effects of the crude aqueous extract from Cecropia pachystachya leaves in a rat model of mania induced by ketamine. The results indicated that ketamine treatment (25 mg/kg i.p., for 8 days) induced hyperlocomotion in the open-field test and oxidative damage in prefrontal cortex and hippocampus, evaluated by increased lipid peroxidation, carbonyl protein formation and decreased total thiol content. Moreover, ketamine treatment reduced the activity of the antioxidant enzymes superoxide dismutase and catalase in hippocampus. Pretreatment of rats with C. pachystachya aqueous extract (200 and 400 mg/kg p.o., for 14 days) or with lithium chloride (45 mg/kg p.o., for 14 days, used as a positive control) prevented both behavioral and pro-oxidant effects of ketamine. These findings suggest that C. pachystachya might be a useful tool for preventive intervention in bipolar disorder, reducing the episode relapse and the oxidative damage associated with the manic phase of this disorder .

This article seeks to investigate associations between satisfaction with life and sociodemographic variables, health conditions, functionality, social involvement and social support among elderly caregivers and non-caregivers, as well as between satisfaction and the intensity of stress in the caregiver group. A sample of 338 caregivers was selected according to two items of the Brazilian version of the Elders Life Stress Inventory. A comparison-group of elderly non-caregivers was selected at random, with a similar gender, age and income profile. Data were derived from self-reported questionnaires and scales. Elderly caregivers with low levels of satisfaction and high levels of stress revealed more symptoms of insomnia, fatigue, diseases and worse IADL performance. Those with greater satisfaction and less stress revealed a good level of social support. Insomnia, depression and fatigue were associated with low satisfaction among caregivers, and with fatigue, depression and low social support among non-caregivers. It was considered relevant that instrumental, psychological and informative support can improve the quality of life and the quality of care provided by elderly caregivers, especially if they are affected by unfavorable health and psychosocial conditions and low satisfaction with life.

Bipolar disorder (BD) is a chronic and debilitating illness characterized by recurrent manic and depressive episodes. Our research investigates the protective effects of curcumin, the main curcuminoid of the Indian spice turmeric, in a model of mania induced by ketamine administration in rats. Our results indicated that ketamine treatment (25 mg/kg, for 8 days) induced hyperlocomotion in the open-field test and oxidative damage in prefrontal cortex (PFC) and hippocampus (HP), evaluated by increased lipid peroxidation and decreased total thiol content. Moreover, ketamine treatment reduced the activity of the antioxidant enzymes superoxide dismutase and catalase in the HP. Pretreatment of rats with curcumin (20 and 50 mg/kg, for 14 days) or with lithium chloride (45 mg/kg, positive control) prevented behavioral and pro-oxidant effects induced by ketamine. These findings suggest that curcumin might be a good compound for preventive intervention in BD, reducing the episode relapse and the oxidative damage associated with the manic phase of this disorder., (Copyright © 2013 Elsevier B.V. All rights reserved.)

There is increasing evidence that motor imagery involves at least in part central processes used in motor control. In order to deepen our understanding on the neural mechanisms underlying vegetative responses to real and imagined exercise, we determined cardioventilatory variables during actual or imagined treadmill walking on flat terrain at speeds of 2, 3.5 or 5 km/h, in a group of 14 healthy volunteers. During actual walking, as expected, a comparable intensity-dependent increase was found in ventilation, oxygen consumption, tidal volume and respiratory rate. Imagined walking led to a significant, albeit small (less than 10%), increase in ventilation and oxygen consumption, and to larger increases (up to 40%) in respiratory rate, which was paralleled by a non significant trend towards a decline of tidal volume. These results confirm and extend previous observations showing that motor imagery is accompanied by centrally induced changes in vegetative responses, and provide evidence for a differential control on respiratory rate and tidal volume.

Laparoscopic surgery requires a series of procedures, including intraperitoneal CO2 insufflation, which can cause cardiovascular and hemogasanalytic modifications, potentially able to impair cerebral perfusion. The aim of this study was to evaluate changes in cerebral blood flow velocity during laparoscopic cholecystectomy. Eighteen patients undergoing laparoscopic cholecystectomy were studied. Middle cerebral artery blood flow velocity was monitored using transcranial Doppler ultrasonography. Electrical bioimpedance was employed to measure cardiac output, stroke volume and to calculate derived parameters. End-tidal CO2, mean arterial blood pressure, end expiratory anesthetic concentration and O2 saturation were monitored non-invasively. Cerebral artery blood flow velocity increased significantly after CO2 insufflation (p < 0.05) and remained stable. The highest values were reached after CO2 desufflation. A significant reduction in stroke volume and cardiac output (p < 0.05) associated with increased vascular systemic resistances (p < 0.001) was observed soon after CO2 insufflation. The decrease in cardiac output and the increase in vascular systemic resistances remained significant throughout abdominal insufflation. Heart rate and mean arterial pressure remained substantially unchanged with the exception of a significant decrease (p < 0.001) before CO2 insufflation. There was no significant change in end-tidal CO2 during abdominal insufflation. These findings suggest that the cerebrovascular system can undergo adaptive changes during all phases of laparoscopic surgery. However, the extent of cardio- and cerebrovascular variation indicates the need for careful preliminary evaluation of cerebral hemodynamics in patients with vascular disorders before laparoscopic surgery.

Objectives: To assess the prognostic reliability of multimodality-evoked potentials and to evaluate the diagnostic implications and define the limits of these evoked potentials., Setting: An ICU in a university hospital., Design: Prospective clinical study., Patients: Seventy-three severely head-injured patients aged 10 to 75 yrs., Methods: Serial recording of brainstem auditory-evoked potentials and somatosensory-evoked potentials between days 1 and 21 after trauma. Comparison between evoked potential findings and prognosis, along with clinical data., Results: Considering the single recordings of both brainstem auditory-evoked potentials and somatosensory-evoked potentials, the accuracy of prognostication in predicting a bad outcome was good only for severely abnormal brainstem auditory-evoked potentials. Serial brainstem auditory-evoked potential recordings and simultaneous recordings of brainstem auditory-evoked potentials and somatosensory-evoked potentials proved to be good prognostic indices in predicting a favorable outcome. Brainstem auditory-evoked potentials correlated well with brainstem reflexes and with pupil asymmetries but did not correlate with Glasgow Coma Scale scores., Conclusions: Serial recording and the use of a multimodality approach provided the best prognostic capabilities. The main diagnostic implications were: a) the possibility of detecting brainstem compression by means of brainstem auditory-evoked potentials before the appearance of pupil abnormalities; b) the usefulness of brainstem auditory-evoked potentials in monitoring brainstem function in patients undergoing high-dose barbiturate therapy. The main limitations of evoked potentials were the occurrence of peripheral acoustic damage, the electromagnetic sources of artifacts in the ICU, and the administration of ototoxic drugs.