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Abstract Animals’ ability to orient and navigate relies on selecting an appropriate motor response based on the perception and integration of the environmental information. This is the case, for instance, of the optokinetic response (OKR) in Drosophila melanogaster, where optic flow visual stimulation modulates head movements. Despite a large body of literature on the OKR, there is still a limited understanding, in flies, of the impact on OKR of concomitant, and potentially conflicting, inputs. To evaluate the impact of this multimodal integration, we combined in D. melanogaster, while flying in a tethered condition, the optic flow stimulation leading to OKR with the simultaneous presentation of olfactory cues, based on repellent or masking compounds typically used against noxious insect species. First, this approach allowed us to directly quantify the effect of several substances and of their concentration on the dynamics of the flies’ OKR in response to moving gratings by evaluating the number of saccades and the velocity of the slow phase. Subsequently, this analysis was capable of easily revealing the actual effect, i.e. masking vs. repellent, of the compound tested. In conclusion, we show that D. melanogaster, a cost-affordable species, represents a viable option for studying the effects of several compounds on the navigational abilities of insects.

Abstract Influenza-like illness (ILI) patients co-detected with respiratory pathogens exhibit poorer health outcomes than those with single infections. To address the paucity of knowledge concerning the incidence of concurrent respiratory pathogens, their relationships, and the clinical differences between patients detected with single and multiple pathogens, we performed an in-depth characterization of the oropharyngeal samples of primary care patients collected in Genoa (Northwest Italy), during winter seasons 2018/19–2019/20. The apriori algorithm was employed to evaluate the incidence of viral, bacterial, and viral-bacterial pairs during the study period. The grade of correlation between pathogens was investigated using the Phi coefficient. Factors associated with viral, bacterial or viral-bacterial co-detection were assessed using logistic regression. The most frequently identified pathogens included influenza A, rhinovirus, Haemophilus influenzae and Streptococcus pneumoniae. The highest correlations were found between bacterial-bacterial and viral-bacterial pairs, such as Haemophilus influenzae-Streptococcus pneumoniae, adenovirus-Haemophilus influenzae, adenovirus-Streptococcus pneumoniae, RSV-A-Bordetella pertussis, and influenza B Victoria-Bordetella parapertussis. Viruses were detected together at significantly lower rates. Notably, rhinovirus, influenza, and RSV exhibited significant negative correlations with each other. Co-detection was more prevalent in children aged

Abstract Background Metabolic syndrome (MetS) is a cluster of medical conditions and risk factors correlating with insulin resistance that increase the risk of developing cardiometabolic health problems. The specific criteria for diagnosing MetS vary among different medical organizations but are typically based on the evaluation of abdominal obesity, high blood pressure, hyperglycemia, and dyslipidemia. A unique, quantitative and independent estimation of the risk of MetS based only on quantitative biomarkers is highly desirable for the comparison between patients and to study the individual progression of the disease in a quantitative manner. Methods We used NMR-based metabolomics on a large cohort of donors (n = 21,323; 37.5% female) to investigate the diagnostic value of serum or serum combined with urine to estimate the MetS risk. Specifically, we have determined 41 circulating metabolites and 112 lipoprotein classes and subclasses in serum samples and this information has been integrated with metabolic profiles extracted from urine samples. Results We have developed MetSCORE, a metabolic model of MetS that combines serum lipoprotein and metabolite information. MetSCORE discriminate patients with MetS (independently identified using the WHO criterium) from general population, with an AUROC of 0.94 (95% CI 0.920–0.952, p

Abstract The receptor-binding site of influenza A virus hemagglutinin partially overlaps with major antigenic sites and constantly evolves. In this study, we observe that mutations G186D and D190N in the hemagglutinin receptor-binding site have coevolved in two recent human H3N2 clades. X-ray crystallography results show that these mutations coordinately drive the evolution of the hemagglutinin receptor binding mode. Epistasis between G186D and D190N is further demonstrated by glycan binding and thermostability analyses. Immunization and neutralization experiments using mouse and human samples indicate that the evolution of receptor binding mode is accompanied by a change in antigenicity. Besides, combinatorial mutagenesis reveals that G186D and D190N, along with other natural mutations in recent H3N2 strains, alter the compatibility with a common egg-adaptive mutation in seasonal influenza vaccines. Overall, our findings elucidate the role of epistasis in shaping the recent evolution of human H3N2 hemagglutinin and substantiate the high evolvability of its receptor-binding mode.

Abstract Background Mindfulness practice has gained interest in the management of Chronic Migraine associated with Medication Overuse Headache (CM-MOH). Mindfulness is characterized by present-moment self-awareness and relies on attention control and emotion regulation, improving headache-related pain management. Mindfulness modulates the Default Mode Network (DMN), Salience Network (SN), and Fronto-Parietal Network (FPN) functional connectivity. However, the neural mechanisms underlying headache-related pain management with mindfulness are still unclear. In this study, we tested neurofunctional changes after mindfulness practice added to pharmacological treatment as usual in CM-MOH patients. Methods The present study is a longitudinal phase-III single-blind Randomized Controlled Trial (MIND-CM study; NCT03671681). Patients had a diagnosis of CM-MOH, no history of neurological and severe psychiatric comorbidities, and were attending our specialty headache centre. Patients were divided in Treatment as Usual (TaU) and mindfulness added to TaU (TaU + MIND) groups. Patients underwent a neuroimaging and clinical assessment before the treatment and after one year. Longitudinal comparisons of DMN, SN, and FPN connectivity were performed between groups and correlated with clinical changes. Vertex-wise analysis was performed to assess cortical thickness changes. Results 177 CM-MOH patients were randomized to either TaU group or TaU + MIND group. Thirty-four patients, divided in 17 TaU and 17 TaU + MIND, completed the neuroimaging follow-up. At the follow-up, both groups showed an improvement in most clinical variables, whereas only TaU + MIND patients showed a significant headache frequency reduction (p = 0.028). After one year, TaU + MIND patients showed greater SN functional connectivity with the left posterior insula (p-FWE = 0.007) and sensorimotor cortex (p-FWE = 0.026). In TaU + MIND patients only, greater SN-insular connectivity was associated with improved depression scores (r = -0.51, p = 0.038). A longitudinal increase in cortical thickness was observed in the insular cluster in these patients (p = 0.015). Increased anterior cingulate cortex thickness was also reported in TaU + MIND group (p-FWE = 0.02). Conclusions Increased SN-insular connectivity might modulate chronic pain perception and the management of negative emotions. Enhanced SN-sensorimotor connectivity could reflect improved body-awareness of painful sensations. Expanded cingulate cortex thickness might sustain improved cognitive processing of nociceptive information. Our findings unveil the therapeutic potential of mindfulness and the underlying neural mechanisms in CM-MOH patients. Trial Registration Name of Registry; MIND-CM study; Registration Number ClinicalTrials.gov identifier: NCT0367168; Registration Date: 14/09/2018

ABSTRACT: Objectives: In order to support policymakers in allocating resources, we aimed to assess vaccine effectiveness (VE) of inactivated influenza vaccines (IIVs) available for Italian adults in the 2023/2024 season. Methods: A hospital-based test-negative case-control study was conducted in Genoa between mid-October 2023 and mid-April 2024. Adult (≥18 years) inpatients with prescription of a polymerase chain reaction test for influenza were eligible. Results: Of 1,664 adults analyzed, most (82%) of which were ≥65 years, 114 (6.9%) tested positive for influenza A. Most (92%) cases were caused by subclades 6B.1A.5a.2a and 6B.1A.5a.2a.1 of the A(H1N1)pdm09 subtype. In older adults aged ≥65 years vaccination was effective at 51% (95% CI: 8%, 74%) against any influenza A and 49% (95% CI: 2%, 73%) against A(H1N1)pdm09. Compared with non-vaccinated older adults, VE point estimates for the adjuvanted and, especially, high-dose IIVs were higher than those for the standard-dose non-adjuvanted IIV. Conclusion: The 2023/2024 seasonal influenza vaccination proved moderately effective in preventing hospitalization for laboratory-confirmed influenza. Being more appropriate for older adults, local policymakers and vaccinating physicians should maximize adoption of the enhanced IIVs.

Commuter satisfaction with their chosen mode of transport significantly affects quality of life, well-being, and the sustainability of commuting practices. This study uses a fuzzy-hybrid TOPSIS approach, applied to data from seven functional urban areas in Central Europe. The method effectively creates a composite indicator for assessing satisfaction levels by considering factors such as socioeconomic conditions, geographical features, and mobility-related data. The research analyzes how satisfaction varies according to these factors. Results indicate that commuter satisfaction levels vary across different factors, including age, education, occupation, gender, travel distance, travel time, commuting cost, and income. Notably, those with shorter commuting times and lower expenses tend to report higher satisfaction, while unemployed and low-income commuters, as well as those using active modes of transport, often express lower satisfaction levels. Interestingly, bus riders find their trip more satisfactory than car drivers, but train commuters are the least satisfied of all. Overall, this study provides valuable insights into commuter satisfaction, informing the development of policies and strategies to improve the transport infrastructure and services and promote the choice of sustainable modes.

Abstract Background Methylation of serotonin-related genes has been proposed as a plausible gene-by-environment link which may mediate environmental stress, depressive and anxiety symptoms. DNA methylation is often measured in blood cells, but little is known about the association between this peripheral epigenetic modification and brain serotonergic architecture. Here, we evaluated the association between whole-blood-derived methylation of four CpG sites in the serotonin transporter (SLC6A4) and six CpG sites of the tryptophan hydroxylase 2 (TPH2) gene and in-vivo brain levels of serotonin transporter (5-HTT) and serotonin 4 receptor (5-HT4) in a cohort of healthy individuals (N = 254) and, for 5-HT4, in a cohort of unmedicated patients with depression (N = 90). To do so, we quantified SLC6A4/TPH2 methylation using bisulfite pyrosequencing and estimated brain 5-HT4 and 5-HTT levels using positron emission tomography. In addition, we explored the association between SLC6A4 and TPH2 methylation and measures of early life and recent stress, depressive and anxiety symptoms on 297 healthy individuals. Results We found no statistically significant association between peripheral DNA methylation and brain markers of serotonergic neurotransmission in patients with depression or in healthy individuals. In addition, although SLC6A4 CpG2 (chr17:30,236,083) methylation was marginally associated with the parental bonding inventory overprotection score in the healthy cohort, statistical significance did not remain after accounting for blood cell heterogeneity. Conclusions We suggest that findings on peripheral DNA methylation in the context of brain serotonin-related features should be interpreted with caution. More studies are needed to rule out a role of SLC6A4 and TPH2 methylation as biomarkers for environmental stress, depressive or anxiety symptoms.

Knee stiffness is a widely known and worrying condition in several postoperative knees. Less is known about native stiff knee. The aim of this manuscript is to summarize the available literature on native stiff knee epidemiology, classification and treatment. In 1989 stiff knee was defined as a knee with less than 50° of total range of motion. If range of motion is

Abstract Objective In elderly people loneliness represents a risk factor for dementia and may negatively impact on mental and physical health. The specific contribute of loneliness to cognitive and behavioral functioning have not yet been determined in amyotrophic lateral sclerosis (ALS). Our hypothesis was that loneliness may be related to motor dysfunction with a negative impact on cognitive and behavioral decline, possibly related to specific cortical involvement. Methods In 200 ALS patients (ALSpts) and 50 healthy controls (HCs) we measured loneliness, mood, and quality of life (QoL). ALSpts underwent comprehensive clinical, genetic, and neuropsychological assessment to define phenotypes. Seventy‐seven ALSpts performed 3T MRI scans to measure cortical thickness. Between‐group, partial correlation and regression analyses were used to examined clinical, neuropsychological, and cortical signatures of loneliness. Results Feelings of loneliness were documented in 38% of ALSpts (ALS/L+pts) and in 47% of HCs. In both groups loneliness was associated with anxiety (P

Abstract Most of the patients affected by neuronopathic forms of Mucopolysaccharidosis type II (MPS II), a rare lysosomal storage disorder caused by defects in iduronate-2-sulfatase (IDS) activity, exhibit early neurological defects associated with white matter lesions and progressive behavioural abnormalities. While neuronal degeneration has been largely described in experimental models and human patients, more subtle neuronal pathogenic defects remain still underexplored. In this work, we discovered that the axon guidance receptor Deleted in Colorectal Cancer (Dcc) is significantly dysregulated in the brain of ids mutant zebrafish since embryonic stages. In addition, thanks to the establishment of neuronal-enriched primary cell cultures, we identified defective proteasomal degradation as one of the main pathways underlying Dcc upregulation in ids mutant conditions. Furthermore, ids mutant fish-derived primary neurons displayed higher levels of polyubiquitinated proteins and P62, suggesting a wider defect in protein degradation. Finally, we show that ids mutant larvae display an atypical response to anxiety-inducing stimuli, hence mimicking one of the characteristic features of MPS II patients. Our study provides an additional relevant frame to MPS II pathogenesis, supporting the concept that multiple developmental defects concur with early childhood behavioural abnormalities.

The measurement of the meshing stiffness in gear pairs is a technological problem. Many studies have been conducted, but a few results are available. A tailored test bench was designed and realized to measure the Static Transmission Error in two mating gears to address this issue. The bench is capable of testing several kinds of gears, e.g., spur, helical, conical, and internal, and it measures the transmission error concerning the applied torque. The Static Transmission Error is due to the variable stiffness of the gear teeth during a mesh cycle. In this paper, a dynamical method for measuring gear mesh stiffness is presented. The tooth stiffness is estimated from the torsional modal behavior of the rotating parts of the test bench. The dynamics of the system are acquired using accelerometers and very precise encoders to measure the angular accelerations and displacements of rotating parts. The torsional mode shapes are identified; those that show a vibrational behavior of the gears that do not follow the transmission ratio’s sign of the mating kinematic condition are selected because they depend on the flexibility of the teeth. In such a way, the engagement stiffness is estimated from the natural frequencies of the selected mode-shapes and the known inertia of gears and shafts. The experimentally identified results are also compared with numerical values computed with a commercial software for mutual validation.

Abstract Objective The resting‐state functional connectome has not been extensively investigated in amyotrophic lateral sclerosis (ALS) spectrum disease, in particular in relationship with patients' genetic status. Methods Here we studied the network‐to‐network connectivity of 19 ALS patients carrying the C9orf72 hexanucleotide repeat expansion (C9orf72+), 19 ALS patients not affected by C9orf72 mutation (C9orf72−), and 19 ALS‐mimic patients (ALSm) well‐matched for demographic and clinical variables. Results When compared with ALSm, we observed greater connectivity of the default mode and frontoparietal networks with the visual network for C9orf72+ patients (P = 0.001). Moreover, the whole‐connectome showed greater node degree (P

IntroductionWhile pharmacists-led interventions in hypertension have proven effective in high-income countries, their implementation and impact in low- and middle-income countries (LMIC) remain limited. This study assessed the implementation and outcomes of the hypertension program FarmaTeCuida (FTC), which integrated community pharmacies into the public primary care level using information and communication technologies. The study took place during the pandemic in General Pueyrredón, Buenos Aires, Argentina, so modifications to the implementation strategy and expected outcomes were also analyzed.MethodsA mixed-methods study was conducted using the non-adoption, abandonment, scaling-up, dissemination, and sustainability (NASSS) conceptual model. Qualitative in-depth interviews were conducted with key stakeholders using snowball sampling until thematic saturation was achieved. The quantitative approach employed a quasi-experimental, prospective, longitudinal design in a cohort of hypertensive patients enrolled in the FTC program since October 2020 to March 2022. Adoption, access, adherence to follow-up, and blood pressure levels were assessed. Clinical outcomes were compared to a cohort of hypertensive patients attending primary health care centers (PHCCs) in 2021 but not enrolled in the FTC program. Routine data from this cohort was obtained from the municipal health information system (HIS).ResultsOut of 33 PHCCs, 23 adopted the FTC program, but only four collaborated with community pharmacies. A total of 440 patients were recruited, with 399 (91%) enrolled at PHCCs. Hypertension was detected in 63% (279/440) of cases at the first visit (113 were possible hypertensive patients; 26 new hypertensive patients and 140 already diagnosed). During follow-up, FTC identified 52 new hypertensive patients (12% out of 440). Reduction of systolic blood pressure (SBP) was observed in patients enrolled in both the FTC program and the comparison group over 60 days. In the multivariate analysis that included all hypertensive patient (FTC and HIS) we found strong evidence that for each month of follow up, SBP was reduced by 1.12 mmHg; however, we did not find any significant effect of the FTC program on SBP trend (interaction FTC*months has a p-value = 0.23). The pandemic was identified as the main reason for the program's underperformance; in addition we identified barriers related to technology, patient suitability, implementation team characteristics, and organizational factors.DiscussionOur study, grounded in the NASSS model, highlights the profound complexity of introducing innovative strategies in low- and middle-income settings. Despite substantial challenges posed by the pandemic, these obstacles provided valuable insights, identified areas for improvement, and informed strategies essential for reshaping the care paradigm for conditions like hypertension in resource-constrained environments.

La evaluación de pastizales en regiones áridas y semiáridas de la Patagonia argentina es una herramienta clave tanto para planificar el manejo ganadero como para realizar evaluaciones ambientales y seguir procesos complejos (e.g., desertificación e impacto del cambio climático). Mientras la mayoría de los métodos de evaluación de pastizales asignan esfuerzos a resolver el dato (parcelas de evaluación a campo) o a la extensión de la información (clasificaciones a escala de paisaje usando sensores remotos), la variación temporal de la vegetación concentra menos atención. Las razones de este déficit podrían responder al costo de realizar evaluaciones a campo en el largo plazo o a las cortas series de datos satelitales disponibles hasta hace dos décadas. Una ventaja actual es contar con series de datos satelitales de >20 años, lo cual posibilita estudiar la dinámica (i.e., la variabilidad temporal y espacio-temporal) de la productividad de la vegetación. Nuestro objetivo fue evaluar la aplicación de un método de clasificación funcional de la vegetación, analizando la complementación de información entre aspectos estructurales y funcionales de ambientes de pastizal a escala de un predio ganadero. Comparamos los resultados obtenidos por una clasificación de unidades de paisaje y fisionomía de la vegetación con los de una clasificación y caracterización de la dinámica de la productividad de la vegetación a partir del protocolo que denominamos Arquetipos Funcionales de Pastizal. Los resultados enfatizan la necesidad de incorporar la dinámica de la productividad vegetal como un factor clasificatorio y no como una variable dependiente de una clasificación estructural definida de manera previa. Discutimos la potencialidad y los pasos futuros en la integración de ambos abordajes a escala predial en sistemas ganaderos de Patagonia como herramienta para la evaluación complementaria estructural y funcional de pastizales, orientada a la planificación y al manejo adaptativo.

Objetivo. Analizar la existencia y actualización de las listas de medicamentos nacionales (LMN) y guías de práctica clínica (GPC) para el tratamiento de la diabetes en América Latina y el Caribe (ALC). Comparar los fármacos incluidos en las listas y guías de cada país, entre sí y con los de la Organización Mundial de la Salud (OMS). Métodos. Estudio de corte transversal. Se identificaron las LMN y GPC para diabetes en los sitios web de la Organización Panamericana de la Salud y de las autoridades sanitarias nacionales. Se relevaron los fármacos y se analizaron por grupo farmacológico según el cuarto nivel de la nomenclatura ATC. Se utilizó el puntaje F1 para evaluar la proximidad de las LMN con la lista modelo de medicamentos esenciales (LMME) de la OMS. Resultados. Del total de países, 87,2% cuentan con LMN, y 91% con GPC (78% y 45% actualizadas en los últimos 5 años, respectivamente). En comparación con los 6 grupos de hipoglucemiantes de la LMME, las LMN tenían una mediana (rango) de 6 (4-13) y un puntaje F1 de 0,80; esto indica una consonancia adecuada. Las GPC tenían una mediana (rango) de 12 (1-12) hipoglucemiantes frente a los 8 de las guías de la OMS. Las GPC tuvieron una mediana de 15 fármacos más que las respectivas LMN. Conclusiones. Si bien la mayoría de los países de ALC cuentan con LMN y GPC para diabetes, la falta de concordancia entre ellas limita su eficacia. Es necesario alinear los procesos y criterios de elaboración de estas dos herramientas de la política de medicamentos.

With the rapid increase of the world population, climate changes, and the slow expansion of cultivated areas, only precision agriculture (PA) can provide enough food or resources. PA requires flexible instruments for measuring the spectral signatures of the crops to understand their conditions. Unfortunately, the high initial costs of multispectral (MS) cameras reduce the implementation of PA in small farms, which constitute a large portion of arable land in Europe and contribute with social bonds, local know-how, and cultural legacy. With the objective to speed up the use of MS imaging, in this article, we present a novel low-cost imaging device consisting of an MS camera with nine bands and a thermal imager, whose price is several times lower than commercially available ones. This article describes the design and the calibration of the imaging device based on the off-the-shelf components: Raspberry Pi, dedicated quad camera kit, thermal core, and multiband optical filters. The spectral reconstruction accuracy has a high average R2 score of 0.986. Finally, images from multiple sensors are aligned using phase-only correlation and dense optical flow, providing a method that can be implemented on all platforms. The presented solution is open source, permitting one to modify and expand the capabilities of the described device and adapt to specific needs. Moreover, the device is flexible as the thermal camera can be removed to reduce the total system cost if its usage is not required. Even if its primary application is PA, the proposed solution can be used for other applications.

Abstract Background Despite hypothalamus has long being considered to be involved in the pathophysiology of cluster headache, the inconsistencies of previous neuroimaging studies and a limited understanding of the hypothalamic areas involved, impede a comprehensive interpretation of its involvement in this condition. Methods We used an automated algorithm to extract hypothalamic subunit volumes from 105 cluster headache patients (57 chronic and 48 episodic) and 59 healthy individuals; after correcting the measures for the respective intracranial volumes, we performed the relevant comparisons employing logist regression models. Only for subunits that emerged as abnormal, we calculated their correlation with the years of illness and the number of headache attacks per day, and the effects of lithium treatment. As a post-hoc approach, using the 7 T resting-state fMRI dataset from the Human Connectome Project, we investigated whether the observed abnormal subunit, comprising the paraventricular nucleus and preoptic area, shows robust functional connectivity with the mesocorticolimbic system, which is known to be modulated by oxytocin neurons in the paraventricular nucleus and that is is abnormal in chronic cluster headache patients. Results Patients with chronic (but not episodic) cluster headache, compared to control participants, present an increased volume of the anterior–superior hypothalamic subunit ipsilateral to the pain, which, remarkably, also correlates significantly with the number of daily attacks. The post-hoc approach showed that this hypothalamic area presents robust functional connectivity with the mesocorticolimbic system under physiological conditions. No evidence of the effects of lithium treatment on this abnormal subunit was found. Conclusions We identified the ipsilateral-to-the-pain antero-superior subunit, where the paraventricular nucleus and preoptic area are located, as the key hypothalamic region of the pathophysiology of chronic cluster headache. The significant correlation between the volume of this area and the number of daily attacks crucially reinforces this interpretation. The well-known roles of the paraventricular nucleus in coordinating autonomic and neuroendocrine flow in stress adaptation and modulation of trigeminovascular mechanisms offer important insights into the understanding of the pathophysiology of cluster headache.

Background and Objectives: The steady spread of dengue virus (DENV) poses a profound public health threat worldwide. Reverse transcription real-time polymerase chain reaction (RT2-PCR) has been increasingly recognized as a reference method for the diagnosis of acute dengue infection. The goal of this study was to assess the diagnostic accuracy of five different RT2-PCR kits for the detection of DENV in a historically processed set of sera samples. Materials and Methods: In this retrospective study, 25 sera samples from routinely processed unique adult patients with a known DENV status (previously tested in both molecular and serological assays) were tested in parallel using four conventional (RealStar Dengue PCR Kit 3.0, Clonit’ngo Zika, Dengue & Chikungunya, BioPerfectus Zika Virus/Dengue Virus/Chikungunya Virus Real Time PCR Kit and Novaplex Tropical fever virus) and one sample-to-result (STANDARD M10 Arbovirus Panel) RT2-PCR assays. Additionally, an end-point dilution analysis was conducted in quintuplicate on six serial dilutions of an RNA preparation obtained from a culture-grown DENV serotype 1 strain for a total of 150 tests. Results: The overall accuracy of the evaluated tests ranged from 84% to 100%. In particular, the sensitivity of three conventional RT2-PCR assays (RealStar, Clonit’ngo and Novaplex) was 100% (95% CI: 79.6–100%), while it was lower (73.3%; 95% CI: 48.1–89.1%) for the BioPerfectus kit. The sample-to-result STANDARD M10 panel performed comparatively well, showing a sensitivity of 92.9% (95% CI: 68.5–98.7%). No false positive results were registered in any assay. The end-point dilution analysis suggested that the RealStar kit had the lowest limit of detection. Conclusions: Available RT2-PCR kits for the detection of DENV are highly specific and generally sensitive and, therefore, their implementation in diagnostic pathways is advisable.

Background: Next-generation sequencing (NGS) is gradually replacing Sanger sequencing for HIV genotypic drug resistance testing (GRT). This work evaluated the concordance among different NGS-GRT interpretation tools in a real-life setting. Methods: Routine NGS-GRT data were generated from viral RNA at 11 Italian laboratories with the AD4SEQ HIV-1 Solution v2 commercial kit. NGS results were interpreted by the SmartVir system provided by the kit and by two online tools (HyDRA Web and Stanford HIVdb). NGS-GRT was considered valid when the coverage was >100 reads (100×) at each PR/RT/IN resistance-associated position listed in the HIVdb 9.5.1 algorithm. Results: Among 629 NGS-GRT, 75.2%, 74.2%, and 70.9% were valid according to SmartVir, HyDRA Web, and HIVdb. Considering at least two interpretation tools, 463 (73.6%) NGS-GRT had a valid coverage for resistance analyses. The proportion of valid samples was affected by viremia 10% showed fair concordance among different interpretation tools. Conclusion: This Italian survey on NGS resistance testing suggests that viremia levels and HIV subtype affect NGS-GRT coverage. Within the current routine method for NGS-GRT, only mutations with frequency >10% seem reliably detected across different interpretation tools.

IntroductionAssessing the treatment response of glioblastoma multiforme during immunotherapy (IT) is an open issue. Treatment response assessment maps (TRAMs) might help distinguish true tumor progression (TTP) and pseudoprogression (PsP) in this setting.MethodsWe recruited 16 naïve glioblastoma patients enrolled in a phase II trial consisting of the Stupp protocol (a standardized treatment for glioblastoma involving combined radiotherapy and chemotherapy with temozolomide, followed by adjuvant temozolomide) plus IT with dendritic cells. Patients were followed up till progression or death; seven underwent a second surgery for suspected progression. Clinical, immunological, and MRI data were collected from all patients and histology in case of second surgery. Patients were classified as responders (progression-free survival, PFS > 12 months), and non-responders (PFS ≤ 12), HIGH-NK (natural killer cells, i.e., immunological responders), and LOW-NK (immunological non-responders) based on immune cell counts in peripheral blood. TRAMs differentiate contrast-enhancing lesions with different washout dynamics into hypothesized tumoral (conventionally blue-colored) vs. treatment-related (red-colored).ResultsUsing receiver operating characteristic (ROC) curves, a threshold of −0.066 in VBlue/VCE (volume of the blue portion of tumoral area/volume of contrast enhancement) variation between values obtained in the MRI performed before PsP/TTP and at TTP/PSP allowed to discriminate TTP from PsP with a sensitivity of 71.4% and a specificity of 100%. Among HIGH-NK patients, at month 6 there was a significant reduction compared to baseline and month 2 in median “blue” volumes.DiscussionIn conclusion, in our pilot study TRAMs support the discrimination between tumoral and treatment-related enhancing features in immunological responders vs. non-responders, the distinction between PsP and TTP, and might provide surrogate markers of immunological response.

ABSTRACT: Objectives: Although respiratory syncytial virus (RSV) is a leading cause of acute respiratory infections (ARIs), it is unclear which of the case definitions that prompt swab collection predicts RSV best. We aimed to profile RSV-positive adults and to identify possible RSV case definitions. Methods: This individual-based pooled analysis was based on influenza-like illness (ILI) surveillance conducted among Italian outpatient adults. All samples were tested for influenza, RSV and other respiratory viruses. Results: RSV was detected in 5.2% of the 1240 ILI adults tested. The prevalence of fever/feverishness was significantly lower (83.3%) in individuals positive for RSV and those negative for both viruses (79.4%) than in influenza-positive subjects (96.2%). Conversely, 98.3% of RSV-positive adults reported cough. Compared with subjects who tested negative, the adjusted relative risk ratio of cough in RSV-positive subjects was much higher than in influenza-positive subjects (6.89 vs 2.79). Using ARI with cough as the RSV case definition increased specificity. Conclusion: As fever/feverishness is more common among influenza than RSV cases, ILI-based surveillance may underestimate RSV incidence in adult outpatients. While broad ARI definitions are useful for routine RSV surveillance, their low specificity may hamper vaccine effectiveness studies. The use of further ARI qualifiers like cough increases specificity.

The cactus moth, Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) is native to South America and has been used as a biocontrol agent of Opuntia (Cactaceae) in Australia and South Africa. Its invasion in North America has raised concerns for the native Opuntia in the USA and Mexico. We investigated the reproductive biology and rearing procedures of a host-specific potential biocontrol agent, Apanteles opuntiarum Martínez and Berta (Hymenoptera: Braconidae). Given the gregarious nature of the parasitoid larvae, we studied the morphology of the immature stages and evaluated evidence of polyembryony and superparasitism. We also investigated the effects of host exposure arena and host density on parasitism rates and wasp production. The morphological descriptions provide a basis for comparison with other species. Early larval instars of A. opuntiarum are similar to other microgastrine immature stages. However, the mature larva exhibits placoid sensilla in the epistomal region, a unique character not previously reported. We provide evidence that A. opuntiarum eggs are not polyembryonic; females frequently superparasitize and have an oviposition preference for larvae parasitized 1–2 d previously. Exposing larvae of C. cactorum to wasps while inside the cactus resulted in lower parasitism and fewer offspring from each host than exposing larvae in the arena without the cactus. Parasitism and mortality rates were higher at lower host densities, possibly due to reduced host group defensive behavior. These results suggest that preference for superparasitism, host defensive behavior, and interactions with the cactus may play an important role in per-host wasp production under laboratory conditions.

This article introduces the Lake Environmental Data Harvester (LED) System, a robotic platform designed for the development of an innovative solution for monitoring remote alpine lakes. LED is intended as the first step in creating portable robotic tools that are lightweight, cost-effective, and highly reliable for monitoring remote water bodies. The LED system is based on the Shallow-Water Autonomous Multipurpose Platform (SWAMP), a groundbreaking Autonomous Surface Vehicle (ASV) originally designed for monitoring wetlands. The objective of LED is to achieve the comprehensive monitoring of remote lakes by outfitting the SWAMP with a suite of sensors, integrating an IoT infrastructure, and adhering to FAIR principles for structured data management. SWAMP’s modular design and open architecture facilitate the easy integration of payloads, while its compact size and construction with a reduced weight ensure portability. Equipped with four azimuth thrusters and a flexible hull structure, SWAMP offers a high degree of maneuverability and position-keeping ability for precise surveys in the shallow waters that are typical of remote lakes. In this project, SWAMP was equipped with a suite of sensors, including a single-beam dual-frequency echosounder, water-quality sensors, a winch for sensor deployment, and AirQino, a low-cost air quality analysis system, along with an RTK-GNSS (Global Navigation Satellite System) receiver for precise positioning. Utilizing commercial off-the-shelf (COTS) components, a Multipurpose Data-Acquisition System forms the basis for an Internet of Things (IoT) infrastructure, enabling data acquisition, storage, and long-range communication. This data-centric system design ensures that acquired variables from both sensors and the robotic platform are structured and managed according to the FAIR principles.

IntroductionDuring thermogenesis, adipose tissue (AT) becomes more active and enhances oxidative metabolism. The promotion of this process in white AT (WAT) is called “browning” and, together with the brown AT (BAT) activation, is considered as a promising approach to counteract obesity and metabolic diseases. Transient receptor potential cation channel, subfamily M, member 2 (TRPM2), is an ion channel that allows extracellular Ca2+ influx into the cytosol, and is gated by adenosine diphosphate ribose (ADPR), produced from NAD+ degradation. The aim of this study was to investigate the relevance of TRPM2 in the regulation of energy metabolism in BAT, WAT, and liver during thermogenesis.MethodsWild type (WT) and Trpm2-/- mice were exposed to 6°C and BAT, WAT and liver were collected to evaluate mRNA, protein levels and ADPR content. Furthermore, O2 consumption, CO2 production and energy expenditure were measured in these mice upon thermogenic stimulation. Finally, the effect of the pharmacological inhibition of TRPM2 was assessed in primary adipocytes, evaluating the response upon stimulation with the β-adrenergic receptor agonist CL316,243.ResultsTrpm2-/- mice displayed lower expression of browning markers in AT and lower energy expenditure in response to thermogenic stimulus, compared to WT animals. Trpm2 gene overexpression was observed in WAT, BAT and liver upon cold exposure. In addition, ADPR levels and mono/poly-ADPR hydrolases expression were higher in mice exposed to cold, compared to control mice, likely mediating ADPR generation.DiscussionOur data indicate TRPM2 as a fundamental player in BAT activation and WAT browning. TRPM2 agonists may represent new pharmacological strategies to fight obesity.

Sirtuins (SIRTs) are classified as class III histone deacetylases (HDACs), a family of enzymes that catalyze the removal of acetyl groups from the ε-N-acetyl lysine residues of histone proteins, thus counteracting the activity performed by histone acetyltransferares (HATs). Based on their involvement in different biological pathways, ranging from transcription to metabolism and genome stability, SIRT dysregulation was investigated in many diseases, such as cancer, neurodegenerative disorders, diabetes, and cardiovascular and autoimmune diseases. The elucidation of a consistent number of SIRT–ligand complexes helped to steer the identification of novel and more selective modulators. Due to the high diversity and quantity of the structural data thus far available, we reviewed some of the different ligands and structure-based methods that have recently been used to identify new promising SIRT1/2 modulators. The present review is structured into two sections: the first includes a comprehensive perspective of the successful computational approaches related to the discovery of SIRT1/2 inhibitors (SIRTIs); the second section deals with the most interesting SIRTIs that have recently appeared in the literature (from 2017). The data reported here are collected from different databases (SciFinder, Web of Science, Scopus, Google Scholar, and PubMed) using “SIRT”, “sirtuin”, and “sirtuin inhibitors” as keywords.

This paper is focused on the design and development of the Porcospino Flex, a single-track robot inspired by nature and featuring a meta-material structure. In the earlier version of the Porcospino, the main body was composed of a chain of vertebrae and two end sections linked by flexible joints, but the excessive use of materials in 3D printing and the resulting weight of the robot posed challenges, ultimately leading to a decrease in its overall efficiency and performance. The Porcospino Flex is manufactured through the fused deposition modeling process using acrylonitrile butadiene styrene and thermoplastic polyurethane, featuring a singular meta-material structure vertebral column. The adoption of a lattice structure in the main body of the Porcospino Flex leads to a substantial increase in performance, reducing its weight from 4200 g to 3600 g. Furthermore, the decrease in weight leads to a reduction in material usage and waste, making a substantial contribution to the sustainability of the robot. The discussion focuses on the testing results of the Porcospino Flex prototype, highlighting the enhancements observed compared to its prior version.

Background: Pneumocytis jirovecii pneumonia (PJP) has high mortality rates in immunocompromised children, even though routine prophylaxis has decreased in incidence. The aim of this case series is to present the radiological and clinical pathway of PJP in a pediatric population. Description of Cases: All PJP cases in non-HIV/AIDS patients diagnosed at Istituto Giannina Gaslini Pediatric Hospital in Genoa (Italy) from January 2012 until October 2022 were retrospectively evaluated. Nine cases were identified (median age: 8.3 years), and of these, 6/9 underwent prophylaxis with trimethoprim/sulfamethoxazole (TMP/SMX; five once-a-week schedules and one three times-a-week schedule), while 3/9 did not receive this. PJP was diagnosed by real-time PCR for P. jirovecii-DNA in respiratory specimens in 7/9 cases and two consecutive positive detections of β-d-glucan (BDG) in the serum in 2/9 cases. Most patients (6/8) had a CT scan with features suggestive of PJP, while one patient did not undergo a scan. All patients were treated with TMP/SMX after a median time from symptoms onset of 3 days. In 7/9 cases, empirical TMP/SMX treatment was initiated after clinical suspicion and radiological evidence and later confirmed by microbiological data. Clinical improvement with the resolution of respiratory failure and 30-day survival included 100% of the study population. Discussion: Due to the difficulty in obtaining biopsy specimens, PJP diagnosis is usually considered probable in most cases. Moreover, the severity of the clinical presentation often leads physicians to start TMP/SMX treatment empirically. BDG proved to be a useful tool for diagnosis, and CT showed good accuracy in identifying typical patterns. In our center, single-day/week prophylaxis was ineffective in high-risk patients; the three-day/week schedule would, therefore, seem preferable and, in any case, should be started promptly in all patients who have an indication of pneumonia.

Although graph convolutional networks have found application in polarimetric synthetic aperture radar (PolSAR) image classification tasks, the available approaches cannot operate on multiple graphs, which hinders their potential to generalize effective feature representations across different datasets. To overcome this limitation and achieve robust PolSAR image classification, this paper proposes a novel end-to-end cross-level interaction graph U-Net (CLIGUNet), where weighted max-relative spatial convolution is proposed to enable simultaneous learning of latent features from batch input. Moreover, it integrates weighted adjacency matrices, derived from the symmetric revised Wishart distance, to encode polarimetric similarity into weighted max-relative spatial graph convolution. Employing end-to-end trainable residual transformers with multi-head attention, our proposed cross-level interactions enable the decoder to fuse multi-scale graph feature representations, enhancing effective features from various scales through a deep supervision strategy. Additionally, multi-scale dynamic graphs are introduced to expand the receptive field, enabling trainable adjacency matrices with refined connectivity relationships and edge weights within each resolution. Experiments undertaken on real PolSAR datasets show the superiority of our CLIGUNet with respect to state-of-the-art networks in classification accuracy and robustness in handling unknown imagery with similar land covers.

In 2022–23, the human monkeypox virus (MPXV) caused a global outbreak in several non-endemic countries. Here, we evaluated the diagnostic performance of four real-time qualitative PCR assays for the laboratory diagnosis of mpox (monkeypox) monkeypox disease. From July to August 2022, 27 positive and 10 negative specimens (lesion, crust and exudate swabs) were tested in the laboratory of the Hygiene Unit of the San Martino Hospital (Genoa, Italy) by using home-made real-time PCR to detect MPXV generic G2R_G DNA. According to the manufacturer’s instructions, we also retrospectively analyzed these specimens using RealCycler MONK-UX/-GX (Progenie Molecular), STANDARD M10 MPX/OPX (SD Biosensor), Novaplex MPXV (Seegene Inc.) and RealStar Orthopoxvirus PCR Kit 1.0 (Altona Diagnostics) assays, recognized as research-use-only tests. The diagnostic accuracy and sensitivity of these assays ranged from 97.3% (95% CI: 86.2–99.5%) to 100% (95% CI: 90.6–100%) and 96.3% (95% CI: 81.72–99.34%) to 100% (95% CI: 72.2–100%), respectively. The RealCycler MONK-UX and STANDARD M10 MPX/OPX did not detect one positive sample with a cycle threshold of 36. The overall specificity was 100% (95% CI: 72.2–100%), and Cohen’s Kappa values ranged from 1 (95% CI: 0.67–1) to 0.93 (95% CI: 0.61–1). As they are highly accurate, reliable and user-friendly, these tests should be recommended for the routine or rapid laboratory discrimination of mpox from other rash illnesses.

The effects of an early impoverished social or physical environment on vertebrate neural development and cognition has been known for decades. While existing studies have focused on the long-term effects, measuring adult cognitive phenotypes, studies on the effects of environmental complexity on the early stages of development are lacking. Zebrafish (Danio rerio) hatchlings are assumed to have minimal interaction with their environment and are routinely reared in small, bare containers. To investigate the effects of being raised under such conditions on development of behaviour and cognition, hatchlings housed for 10 days in either an enriched or a standard environment underwent two cognitive tasks. The results were mixed. Subjects of the two treatments did not differ in performance when required to discriminate two areas. Conversely, we found a significant effect in a number discrimination task, with subjects from impoverished condition performing significantly worse. In both experiments, larvae reared in impoverished environment showed a reduced locomotor activity. Given the effects that enrichment appears to exert on larvae, a third experiment explored whether hatchlings exhibit a spontaneous preference for more complex environments. When offered a choice between a bare setting and one with objects of different shapes and colors, larvae spent over 70% of time in the enriched sector. Deepening these effects of an early impoverished environment on cognitive development is crucial for the welfare of captive zebrafish populations and for enhancing the quality and reliability of studies on larval zebrafish.

To quantitatively evaluate the possible advantages of quantifying and differentiating various soft tissues using virtual monochromatic images (VMI) derived from different dual-energy computed tomography (DECT) technologies. This study involved four DECT scanners with different technologies. CIRS phantom images were acquired in single-energy (SECT) and DECT modes with each scanner. The analysis focused on five equivalent soft-tissue inserts: adipose, breast, liver, muscle, and bone (200 mg). The signal-to-noise ratio (SNR) was calculated for each equivalent soft-tissue insert. Finally, the contrasts of tissue pairs between DECT and SECT images were compared using Wilcoxon signed-rank tests adjusted for multiple comparisons. Average CT numbers and noise showed a significant difference pattern between DECT with respect to SECT for each CT scanner. Generally, energy levels of 70 keV or higher led to improved SNR in VMI for most of the equivalent soft-tissue inserts. However, energy levels of 40–50 keV showed significantly higher contrasts in most of the equivalent soft-tissue insert pairs. DECT images at low energies, especially at 40–50 keV, outperform SECT images in discriminating soft tissues across all four DECT technologies. The combined use of DECT images reconstructed at different energy levels provides a more comprehensive set of information for diagnostic and/or radiotherapy evaluation compared to SECT. Some differences between scanners are evident, depending on the DECT acquisition technique and reconstruction method.

The Yangtze River Basin (YZRB) and the Yellow River Basin (YRB), which are crucial for ecology and economy in China, face growing challenges to ecosystem service (ES) functions due to global population growth, urbanization, and climate change. This study assessed the spatiotemporal dynamics of ESs in the YZRB and the YRB between 2001 and 2021, comprehensively encompassing essential aspects such as water yield (WY), carbon sequestration (CS), soil conservation (SC), and habitat quality (HQ) while also analyzing the trade-offs and synergies among these ESs at the grid cells. The GeoDetector was employed to ascertain individual or interactive effects of natural and anthropogenic factors on these ESs and their trade-offs/synergies. The results showed that (1) from 2001 to 2021, the four ESs exhibited significant spatial disparities in the distribution within two basins, with the overall trend of ESs mainly increasing. YZRB consistently exhibited substantially higher ES values than the YRB. (2) Complex trade-offs and synergies among these ESs were apparent in both basins, characterized by distinct spatial heterogeneity. The spatial relationships of WY–CS, WY–SC, CS–SC, and CS–HQ were mainly synergistic. (3) Precipitation, potential evapotranspiration, elevation, land use and land cover (LULC), and slope influenced ESs in both basins. Notably, interactive factors, particularly the interactions involving LULC and other factors, demonstrated more robust explanatory power for ESs and their trade-offs/synergies than individual drivers. These findings significantly affect the refined ecosystem management and sustainable development decision-making in large rivers or regions.