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Woody biomass represents an important source of carbon on earth, and its global recycling is highly dependent on Agaricomycetes fungi. White-rot Basidiomycetes are a very important group in this regard, as they possess a large and diverse enzymatic repertoire for biomass decomposition. Among these enzymes, the recently discovered lytic polysaccharide monooxygenases (LPMOs) have revolutionized biomass processing with their novel oxidative mechanism of action. The strikingly high representation of LPMOs in fungal genomes raises the question of their functional versatility. In this work, we studied an AA9 LPMO from the white-rot basidiomycete Pycnoporus sanguineus, PsAA9A. Successfully produced as a recombinant secreted protein in Pichia pastoris, PsAA9A was found to be a C1-specific LPMO active on cellulosic substrates, generating native and oxidized cello-oligosaccharides in the presence of an external electron donor. PsAA9A boosted cellulolytic activity of glysoside hydrolases from families GH1, GH5, and GH6.This study serves as a starting point towards understanding the functional versatility and biotechnological potential of this enzymatic family, highly represented in wood decay fungi, in Pycnoporus genus. KEY POINTS: • PsAA9A is the first AA9 from P. sanguineus to be characterized. • PsAA9A has activity on cellulose, producing C1-oxidized cello-oligosaccharides. • Boosting activity with GH1, GH5, and GH6 was proven.

Small-Cell Lung Cancer (SCLC) is an aggressive neuroendocrine malignancy with a poor prognosis. Here, we focus on the neuroendocrine SCLC subtypes, SCLC-A and SCLC-N, whose transcription addiction was driven by ASCL1 and NEUROD1 transcription factors which target E-box motifs to activate up to 40% of total genes, the promoters of which are maintained in a steadily open chromatin environment according to ATAC and H3K27Ac signatures. This leverage is used by the marine agent lurbinectedin, which preferentially targets the CpG islands located downstream of the transcription start site, thus arresting elongating RNAPII and promoting its degradation. This abrogates the expression of ASCL1 and NEUROD1 and of their dependent genes, such as BCL2, INSM1, MYC, and AURKA, which are responsible for relevant SCLC tumorigenic properties such as inhibition of apoptosis and cell survival, as well as for a part of its neuroendocrine features. In summary, we show how the transcription addiction of these cells becomes their Achilles's heel, and how this is effectively exploited by lurbinectedin as a novel SCLC therapeutic endeavor.

Background: In international cooperation projects that are carried out in less developed and developing countries, a large number of children with disabilities present cerebral palsy (CP). Orthopaedic disorders are frequent complications associated with this disorder. Their prevention and early intervention are essential to achieve an appropriate therapeutic approach for children with PC and to improve their quality of life. Objective: To describe the treatment approach that is currently used in international cooperation projects for the rehabilitation management of the orthopaedic disorders in children with cerebral palsy. Methods: This is an observational, descriptive, cross-sectional study, carried out by means of an online questionnaire to professionals in the field of Physiotherapy and Rehabilitation working in international cooperation projects. The inclusion criteria were professionals working in the rehabilitation field in development aid, humanitarian action or emergency projects that provided rehabilitation services, working with children with cerebral palsy from 0 to 18 years old. Results: Ninety-eight questionnaires were analysed. The average age of the participants was 33.2 years, they were mainly working in development cooperation projects (83.33%) that were implemented in rehabilitation centres and through community-based rehabilitation services (60%). The projects were located in countries all over the world but mainly on the Asian continent (71.4%). Physiotherapists and orthopaedic technicians (72.22%) were the main professionals working in these projects, followed by occupational therapists and social workers (55.56%). The results indicated that the orthopaedic disorders were very frequent in the sample (66.67%), with hip subluxation (50%), scoliosis (77.78%), kyphosis (61.1%), clubfoot (88.7%) and varus foot (61.11%) standing out. The most commonly used treatment approaches were positioning (88.89%) and the Bobath concept (83.33%). The technical aids that were used by the professionals were ankle foot orthosis (AFO) (94.44%), bracing (66.67%), standing frames (83.33%), moulded seats (100%), corner seats (93.75%) and adapted seats (92.85%). Conclusions: In international cooperation projects, the rehabilitation treatment of children with cerebral palsy is based on a holistic approach. This is reflected in the interventions that are carried out to treat their orthopaedic disorders and in locally produced devices, awareness raising and community education. However, the professionals surveyed considered that the aids or orthoses used are insufficient in the treatment and prevention of orthopaedic disorders in cerebral palsy.

Objective The study aimed to describe the cases of neurological disease related to the outbreak of enterovirus (EV) in three regions in Northern Spain during 2016. Materials and Methods Multicenter retrospective observational study. Clinical, radiological, and microbiological data were analyzed from patients younger than 15 years with confirmed EV-associated neurological disease admitted to 10 hospitals of Asturias, Cantabria, and Castile and Leon between January 1 and December 31, 2016. Results Fifty-five patients were included. Median age was 24 months (interquartile range = 18.5 months). Fifteen patients were classified as aseptic meningitis (27.3%). In total, 37 cases presented brainstem encephalitis (67.3%), 25 of them due to EV-A71 with excellent prognosis (84.6% asymptomatic 2 months following the onset). Three cases of acute flaccid myelitis (5.5%) by EV-D68 were reported and presented persistent paresis 2 months following the onset. Microbiological diagnosis by reverse transcriptase polymerase chain reaction was performed in all cases, finding EV in cerebrospinal fluid in meningitis, but not in brainstem encephalitis and acute flaccid myelitis, where EV was found in respiratory or rectal samples. Step therapy was administrated with intravenous immunoglobulin (IVIG; 32.7%), methylprednisolone (10%), and plasmapheresis (3.6%). Four patients received fluoxetine (7.3%). Twenty patients needed to be admitted to pediatric intensive care unit (36.4%). Conclusion Clinical, microbiological, and radiological diagnosis is essential in outbreaks of EV neurological disease, taking into account that it can be difficult to identify EV-A71 and EV-D68 in CSF, requiring throat or rectal samples. There is not specific treatment to these conditions and the efficacy and understanding of the mechanism of action of immune-modulatory treatment (IVIG, corticosteroids, and plasmapheresis) is limited.

Treatment of congenital cytomegalovirus infection is mandatory in cases with severe systemic and/or neurological involvement. However, some patients are paucisymptomatic, with very subtle systemic manifestations and/or minimal brain alterations. Current international guidelines do not clearly state whether these children should be treated, and this decision is not straightforward for clinicians. Of a small series of six infants with congenital cytomegalovirus infection admitted to our neonatal unit between 2015 and 2019, half showed paucisymptomatic neurological manifestations. In these cases, the determination of s2-microglobulin in cerebrospinal fluid and magnetic resonance imaging aided in the decision-making concerning the therapeutic approach to follow.

Chronic leg ulcers can have a major impact on the quality of life of patients. These wounds can be complex and hard to heal, as several factors may affect the outcome. Underlying conditions, bacterial growth and excess moisture may prevent wounds from healing. We describe the case of a patient with known chronic venous disease, who was admitted to our hospital for several complex, irregular and infected chronic venous ulcers in his lower legs. The management was frustrating for several months, until we began to use methacrylate powder dressing for his hard-to-heal wounds.

Dihydroceramide desaturase 1 (Des1) catalyzes the last step of ceramide synthesis de novo, thus regulating the physiologically relevant balance between dihydrosphingolipids and sphingolipids. Here we report on the configurational preference of Des1 towards isomeric Δ

A popular method to estimate proximal/distal temperature (TPROX and TDIST) consists in calculating a weighted average of nine wireless sensors placed on pre-defined skin locations. Specifically, TPROX is derived from five sensors placed on the infra-clavicular and mid-thigh area (left and right) and abdomen, and TDIST from four sensors located on the hands and feet. In clinical practice, the loss/removal of one or more sensors is a common occurrence, but limited information is available on how this affects the accuracy of temperature estimates. The aim of this study was to determine the accuracy of temperature estimates in relation to number/position of sensors removed. Thirteen healthy subjects wore all nine sensors for 24 hours and reference TPROX and TDIST time-courses were calculated using all sensors. Then, all possible combinations of reduced subsets of sensors were simulated and suitable weights for each sensor calculated. The accuracy of TPROX and TDIST estimates resulting from the reduced subsets of sensors, compared to reference values, was assessed by the mean squared error, the mean absolute error (MAE), the cross-validation error and the 25th and 75th percentiles of the reconstruction error. Tables of the accuracy and sensor weights for all possible combinations of sensors are provided. For instance, in relation to TPROX, a subset of three sensors placed in any combination of three non-homologous areas (abdominal, right or left infra-clavicular, right or left mid-thigh) produced an error of 0.13°C MAE, while the loss/removal of the abdominal sensor resulted in an error of 0.25°C MAE, with the greater impact on the quality of the reconstruction. This information may help researchers/clinicians: i) evaluate the expected goodness of their TPROX and TDIST estimates based on the number of available sensors; ii) select the most appropriate subset of sensors, depending on goals and operational constraints.

BACKGROUND & AIMS: Sleep preparation/onset are associated with peripheral vasodilatation and a decrease in body temperature. The hyperdynamic syndrome exhibited by patients with cirrhosis may impinge on sleep preparation, thus contributing to their difficulties falling asleep. The aim of this study was the assessment of skin temperature, in relation to sleep-wake patterns, in patients with cirrhosis. METHODS: Fifty three subjects were initially recruited, and 46 completed the study. Of the final 46, twelve were outpatients with cirrhosis, 13 inpatients with cirrhosis, 11 inpatients without cirrhosis and 10 healthy volunteers. All underwent baseline sleep-wake evaluation and blood sampling for inflammatory markers and morning melatonin levels. Proximal/distal skin temperature and their gradient (DPG) were recorded for 24 hours by a wireless device. Over this period subjects kept a sleep-wake diary. RESULTS: Inpatients with cirrhosis slept significantly less well than the other groups. Inpatients and outpatients with cirrhosis had higher proximal temperature and blunted rhythmicity compared to the other groups. Inpatients with/without cirrhosis had higher distal temperature values and blunted rhythmicity compared to the other groups. Inpatients and outpatients with cirrhosis had significantly lower DPG values compared to the other groups, and DPG reached near-zero values several hours later. Significant correlations were observed between temperature and sleep-wake variables and inflammatory markers. CONCLUSIONS: Alterations of distal/proximal skin temperature, their gradient and their time-course were observed in patients with cirrhosis, which may contribute to their sleep disturbances. This article is protected by copyright. All rights reserved.

BACKGROUND: Clamping and cutting of the umbilical cord is the most prevalent of all operations, but the optimal timing of cord clamping is controversial, with different timings offering advantages and disadvantages. This study, for the first time, compares the influence of early and late cord clamping in correlation with oxidative stress and inflammation signaling, Because cord clamping timing may have a significant influence on placenta-to-infant blood transfer, thereby modifying oxygenation of maternal and fetal tissues, and on the transfer of inflammatory mediators throughout the placenta. METHODS: Sixty-four pregnant subjects were selected at the Gynecology and Obstetrics Services Department of the Clinico San Cecilio Hospital, Granada, Spain, based on disease-free women who experienced a normal course of pregnancy and a spontaneous, vaginal, single delivery. Half of the subjects had deliveries with early-clamped newborn infants (at 10 s), and the other half had late-clamped deliveries (at 2 min). RESULTS: Erythrocyte catalase activity was significantly greater in the late-clamped group than in the early-clamped group (P < .01 for the umbilical vein and P < .001 for the artery). The values for superoxide dismutase, total antioxidant status, and soluble tumor necrosis factor receptor II were all significantly higher in the late-clamped group compared with the early-clamped group (P < .01, P < .001, and P < .001, respectively). CONCLUSIONS: The results suggest a beneficial effect of late cord clamping, produced by an increase in antioxidant capacity and moderation of the inflammatory-mediated effects induced during delivery of term neonates.

Little is known about the pathogenesis of cerebral sinovenous thrombosis (CSVT) in the neonate. Although thrombophilia has been described as increasing the risk of CSVT in adults, it remains controversial in pediatric patients, and prospective case–control studies regarding neonatal CSVT are lacking. From 2008 to 2017, all 26 consecutive newborn infants ≥35 weeks of gestation diagnosed with neonatal CSVT, and their mothers, were tested for factor V Leiden (FV) G1691A, FII G20210A, and methylenetetrahydrofolate reductase C677T (MTHFR C677T) mutations. Eighty-five mother–infant pairs were recruited as controls. All infants except 1 with CSVT were suspected due to clinical symptoms, mainly seizures (22/25). Magnetic resonance imaging was performed in 24/26 infants. Heterozygous FV G1691A, FII G20210A, and homozygous MTHFR C677T mutations were present in 1/26, 3/26, and 3/20 infants with CSVT, respectively. FII (odds ratio: 10.96; 95% confidence interval [CI]: 1.09-110.35) and male sex (3.93; 95% CI: 1.43-10.76) were associated with CSVT. When FII G20210A analysis was adjusted for sex, the OR for FII G20210A was 6.70 (95% CI: 0.65-69.22). No differences were found for FV G1691A or homozygous MTHFR mutations between neonates with CSVT and their mothers, compared to controls.

Gender is a crucial determinant of life span, but little is known about gender differences in free radical homeostasis and inflammatory signaling. The aim of the study was to determine gender-related differences concerning oxidative stress and inflammatory signaling of healthy neonates and mothers. Fifty-six mothers with normal gestational course and spontaneous delivery were selected. Blood samples were collected from the mother (at the beginning of delivery and start of expulsive period) and from neonate (from umbilical cord vein and artery). The mothers of girls featured a higher total antioxidant status and lower plasma hydroperoxides than the mother of boys. Regarding the neonates, the girls featured a higher total antioxidant status and lower plasma membrane hydroperoxides in umbilical cord artery together with higher catalase, glutathione peroxidase, and superoxide dismutase activities. Lower levels of interleukin 6, tumor necrosis factor alpha, and prostaglandin E2 were observed in the mothers of girls and higher level of soluble tumor necrosis factor receptor II. In the neonates, lower levels of interleukin 6 and tumor necrosis factor alpha were observed in umbilical artery and higher soluble tumor necrosis factor receptor II in umbilical cord vein and artery of girls. An association between gender, oxidative stress, and inflammation signaling exists, leading to a renewed interest in the neonate’s sex as a potential risk factor to several alterations.

Human papilloma virus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) has a better prognosis than it's HPV negative (HPV(-)) counterpart. This may be due to the higher numbers of tumor-infiltrating lymphocytes (TILs) in HPV positive (HPV(+)) tumors. RNA-Sequencing (RNA-Seq) was used to evaluate whether the differences in clinical behaviour simply reflect a numerical difference in TILs or whether there is a fundamental behavioural difference between TILs in these two settings. Thirty-nine HNSCC tumors were scored for TIL density by immunohistochemistry. After the removal of 16 TILlow tumors, RNA-Seq analysis was performed on 23 TILhigh/med tumors (HPV(+) n=10 and HPV(-) n=13). Using EdgeR, differentially expressed genes (DEG) were identified. Immune subset analysis was performed using Functional Analysis of Individual RNA-Seq/ Microarray Expression (FAIME) and immune gene RNA transcript count analysis. In total, 1,634 DEGs were identified, with a dominant immune signature observed in HPV(+) tumors. After normalizing the expression profiles to account for differences in B- and T-cell number, 437 significantly DEGs remained. A B-cell associated signature distinguished HPV(+) from HPV(-) tumors, and included the DEGs CD200, GGA2, ADAM28, STAG3, SPIB, VCAM1, BCL2 and ICOSLG; the immune signal relative to T-cells was qualitatively similar between TILs of both tumor cohorts. Our findings were validated and confirmed in two independent cohorts using TCGA data and tumor-infiltrating B-cells from additional HPV(+) HNSCC patients. A B-cell associated signal segregated tumors relative to HPV status. Our data suggests that the role of B-cells in the adaptive immune response to HPV(+) HNSCC requires re-assessment.

UNLABELLED Hyperammonaemia/mild hepatic encephalopathy (HE) can be simulated by the oral administration of a so-called amino acid challenge (AAC). This study sought to assess the effects of the AAC alone and in combination with either ammonia-lowering [L-ornithine-L-aspartate (LOLA)] or vigilance-enhancing medication (caffeine). Six patients with cirrhosis (5 males; 61.3 ± 9.2 years; 5 Child A, 1 Child B) and six healthy volunteers (5 males; 49.8 ± 10.6 years) were studied between 08:00 and 19:00 on Monday of three consecutive weeks. The following indices were obtained: hourly capillary ammonia, hourly subjective sleepiness, paper & pencil/computerized psychometry and wake electroencephalography (EEG) at 12:00, i.e. at the time of the maximum expected effect of the AAC. RESULTS On average, patients had worse neuropsychological performance and slower EEG than healthy volunteers in all conditions but differences did not reach significance. In healthy volunteers, the post-AAC increase in capillary ammonia levels was contained by both the administration of LOLA and of caffeine (significant differences between 10:00 and 14:00 h). The administration of caffeine also resulted in a reduction in subjective sleepiness and in the amplitude of the EEG on several frontal/temporal-occipital sites (p

BackgroundApple allergy manifests as two main groups of clinical entities reflecting different patterns of allergen sensitization: oral allergy syndrome (OAS) and generalized symptoms (GS).ObjectiveWe analysed the sensitization profile to a wide panel of different components of food allergens (rMal d 1, Mal d 2, rMal d 3, rMal d 4, rPru p 3, rBet v 1 and Pho d 2) for a population of Mediterranean patients with OAS and GS to apple.MethodsPatients (N = 81) with a history of apple allergy that could be confirmed by positive prick-prick test and/or double-blind-placebo-controlled food challenge (DBPCFC), were included. Skin prick test (SPT) and ELISA were performed using a panel of inhalant, fruit and nut allergens. ELISA and ELISA inhibition studies were performed in order to analyse the sensitization patterns.ResultsThirty-five cases (43.2%) had OAS and 46 (56.8%) GS. SPT showed a significantly higher number of positive results with peach, cherry and hazelnut in those with GS. ELISA showed a significantly high percentage of positive cases to rMal d 3, rMal d 4, rPru p 3 and Pho d 2 in patients with OAS and GS compared to controls, and to rBet v 1 in patients with OAS vs controls and between OAS and GS patients. Three different patterns of recognition were detected: positive to LTP (rMal d 3 or rPru p 3), positive to profilin (rMal d 4 and Pho d 2), or positive to both. There were also patients with rMal d 1 recognition who showed cross-reactivity to rBet v 1.ConclusionIn an apple allergy population with a high incidence of pollinosis different patterns of sensitization may occur. LTP is most often involved in those with GS. Profilin, though more prevalent in patients with OAS, has been shown to sensitise patients with both types of symptoms.

To assess the impact of massage versus ear acupuncture on behavior and participation in occupational therapy of dementia patients.We performed a controlled, randomized longitudinal trial approved by the Bioethics Commission of the University of Extremadura. One hundred twenty elderly subjects with dementia institutionalized in residential homes in Extremadura (Spain) received treatment based on massage and ear acupuncture over three months. Behavior alterations, sleep disturbance, and participation in rehabilitation and eating were assessed every month during the three months of intervention, and at one and two months of follow-up after the end of treatment. The assessment was performed through a structured questionnaire with closed format questions done by an occupational therapist not involved in the study.There was a statistically significant positive effect of massage and ear acupuncture (P0.001) on measured variables in the third month of intervention, which were maintained at two months after completing the treatment (P0.021), when compared to the control group.Massage therapy and ear acupuncture can improve behavior and sleep disturbances, and increase the participation in eating and rehabilitation organized in residential homes, in dementia patients.

This study describes an experience in the training of female shellfish gatherers in the fishing community in Ilha do Paty, São Francisco do Conde, within the context of actions to promote health and food safety. This is an intervention study with planning of activities by a multidisciplinary team. The activities were developed in five stages: survey of topics of interest; awareness; female shellfish gatherers' work routine; teaching of best practices in the processing of shellfish; and group conversation. The methodologies included exposure through dialogue, group dynamics, workshops, theater, work with images, directed activities and the distribution of educational materials. At the end, an evaluation by the participants regarding the different aspects was conducted. Overall there was satisfaction among the audience for most indicators, which may relate to both the use of audiovisual resources and strategies that permitted the exchange of experiences regarding the recognition of the importance of training by the target audience. The experiment demonstrates the potential of developing educational activities with the fishing communities, with results that contribute to the bolstering of the local supply chain, with the promotion of food safety and occupational health.

Food allergy caused by lipid transfer protein (LTP) from peach (Pru p 3) is frequently associated with sensitization to mugwort LTP (Art v 3). Although in vitro cross-reactivity is already well known, it has yet to be elucidated whether a pollen LTP can induce rhinitis.The aim of this study was to investigate whether mugwort LTP could elicit respiratory symptoms and whether a primary food LTP allergy could lead to a respiratory allergy.Patients with confirmed Pru p 3 allergy and control subjects were selected. Immediate responses to nasal allergen provocation tests (NAPTs) with Art v 3, Pru p 3, and mugwort were assessed by using the visual analog scale score, total nasal symptom score, and acoustic rhinometry. Tryptase and cysteinyl leukotriene (cysLT) levels were measured in nasal lavage fluid. Immunoblotting, ELISAs, and ELISA inhibition assays were also performed.Fifteen patients and 9 control subjects were selected. NAPT results with Art v 3 and Pru p 3 showed significant changes in acoustic rhinometry, visual analog scale scores, total nasal symptom scores, and cysLT levels (P.001). Tryptase levels were only increased in NAPTs with Pru p 3. NAPTs with mugwort were used in those patients who were only sensitized to Art v 3, with similar results (P.05). No significant changes were detected in control subjects.The results demonstrated that a pollen LTP can elicit rhinitis in sensitized patients. Findings also suggest that a primary sensitization to Pru p 3 can lead to a respiratory allergy through cross-reactivity.

Objective. The main objective of this study was to systematically review all the scientific studies that analyse the effectiveness of mud pack therapy on patients diagnosed with knee OA. Methods. One hundred and fifteen publications were identified through an electronic and manual search and 20 of those studies were chosen based on the inclusion criteria: randomized clinical trials, systematic reviews or meta-analyses whose objective was to analyse the effect of mud pack therapy on perceived pain, function and quality of life, with a sample size of 520 subjects, published since 2000 and showing conclusive results. Results. In the studies that met the inclusion criteria, 12 analyse functionality, 17 perceived pain, 5 quality of life and all showed a significant improvement in the three analysed variables. The methodological quality of the studies had a moderate risk of bias. Conclusion. Mud pack therapy is considered an alternative and effective therapy in the clinical management of knee OA. Studies with better methodology are needed to prove its scope.

A 100-gram oral glucose tolerance test (OGTT) was performed on a selected group of normal women at each trimester of pregnancy to establish reference values for hyperglycemia and hypoglycemia. Ninety three OGTT were performed in the first trimester, 121 in the second trimester, and 98 in the last trimester. The fasting serum glucose did not differ significantly between the trimesters. The values at 60 and 120 minutes were significantly different for the fifth, fiftieth, and ninety-fifth percentiles between each trimester of pregnancy. For the 180-minute readings, the fifth and fiftieth percentiles were not significantly different between the first and second trimester of pregnancy, but the results of the third trimester were significantly higher than those of the other trimesters. The mean fasting insulinemia remained relatively constant during pregnancy. Insulin response to OGTT increased during the progression of the pregnancy. The interpretation of the glucose tolerance tests during pregnancy, either to detect gestational diabetes or hypoglycemia, should take these physiologic changes into account.