An algorithm for pharmacological treatment of mania during hospitalisation.

Current evidence for pharmacological treatment of mania during hospitalisation is insufficient as there are no larger well-designed randomised trials of comparative medical treatments of mania during inpatient stays. Moreover, there is considerable variation in pharmacological medication in clinical practice during hospitalisation for mania. Based on a hospital data overview, a systematic search of the literature and a three-day consensus meeting, this narrative review proposed an algorithm for optimised pharmacological treatment of mania during hospitalisation and its subsequent scientific evaluation.

Implementing cognitive screenings for outpatients with bipolar disorder following optimised treatment in a specialised mood disorder clinic.

Bipolar disorder (BD) is often accompanied by persistent cognitive impairment. However, screening for cognitive impairment in the clinic is challenged by a lack of consensus on screening procedures. This study assesses cognitive impairment prevalence and screening feasibility in alignment with the International Society for Bipolar Disorder Targeting Cognition Task Force recommendations. Between January 2022 and May 2023, 136 newly diagnosed BD outpatients were assessed with the Screen for Cognitive Impairment in Psychiatry after 15-20 months of specialised care at the Copenhagen Affective Disorder Clinic. Cognitive impairment patterns and associations with cognitive complaints, perceived stress, and functioning were examined. Most screened patients (73 %) achieved full or partial remission, with 51 % being cognitively normal, 38 % showing global impairments, and 11 % displaying selective impairments. Among remitted patients, 56 % were cognitively normal, while 31 % and 13 % exhibited global or selective impairments, respectively. Both objectively impaired patient groups reported more subjective cognitive difficulties than those who were cognitively normal. The globally impaired group also demonstrated poorer functioning, more depressive symptoms and lower quality of life than cognitively normal patients. Across all patients, lower cognitive performance correlated with more cognitive complaints, lower functioning, lower quality of life, and more depressive symptoms. Cognitive screenings were relatively easily implementable, involving only a 1.5 h session including mood ratings, feedback and cognitive strategy discussion. The study highlights the clinical relevance and feasibility of cognitive screenings in BD patients, emphasizing the need for tailored interventions given frequent cognitive impairment in clinically stable individuals.

Convergent and discriminant validity of the Minimal Eating Observation Form - version II: a cross-sectional study.

BACKGROUND: The Minimal Eating Observation Form - Version II (MEOF-II) is a brief and easy to use screening tool for eating difficulties, that is psychometrically robust. The aim of this study was to explore convergent (measuring similar constructs) and discriminant (measuring somewhat different constructs) validity of the MEOF-II to other validated dysphagia specific, activity and participation related instruments. METHODS: In this cross-sectional study, participants (n = 100, mean age 72, n = 42 women), diagnosed with either chronic pulmonary disease, Parkinson´s disease, Multiple Sclerosis, or stroke were recruited from rehabilitation centres. Patient-reported outcomes and clinical-rated assessments, capturing eating ability in general and swallowing in specific, included: The Dysphagia Handicap Index (DHI), the 4-question test (4QT), the Minimal Eating Observation Form - II, the Volume - Viscosity Swallow Test (V-VST), Flexible Endoscopic Evaluation of Swallowing (FEES) documented according to the Penetration-Aspiration Scale (PAS). Type of oral intake was documented using the Functional Oral Intake Scale (FOIS). Activities in daily living was assessed with Barthel index (BI). Spearman's correlation coefficient was used to analyze associations. The MEOF-II total score was hypothesised to have moderate correlations (r ≥ 0.3) with the other assessments, besides with PAS and FOIS (weak correlations, r < 0.3). RESULTS: In total 78 participants had any type of eating difficulties (MEOF-II), 69 reported dysphagia (4QT), 62 had dysphagia according to V-VST, 29 showed evidence of penetration/aspiration (PAS), and 31 participants had decreased oral intake ability (FOIS). The MEOF-II total score had moderate correlations with DHI, BI, 4QT, V-VST volume, and weak correlations with V-VST dysphagia and viscosity, PAS, and FOIS. Comparing a prior hypothesised correlation strengths against empirical findings showed that 83% of the hypothesised correlations were correct. CONCLUSIONS: The MEOF-II is a holistic and objective screening tool that can indicate the need for further assessment and corresponds well with the persons' subjective experiences. MEOF-II does not specifically assess the risk for penetration/aspiration.

Investigation of the SARS-CoV-2 post-vaccination antibody response in Canadian farmed mink.

Currently, SARS-CoV-2 have been detected in farmed mink in 13 different countries. Due to the high susceptibility and transmissibility among mink, great concerns of mink serving as a reservoir to generate novel variants with unknown virulence and antigenic properties arose. These concerns have consequently resulted in entire mink productions being culled and banned. This study investigates the post-vaccination antibody response in the Canadian farmed mink vaccinated with a commercial Index spike protein-based vaccine, approved for use in cats, and compares the antibody response to that observed post infection in Danish farmed mink. Blood samples were obtained from 50 mink at the Canadian Centre for Fur Animal Research (CCFAR), Dalhousie University (Truro, Canada). The sera were initially analyzed for antibodies by enzyme-linked immunosorbent assay (ELISA), and selected sera was subsequently tested in a virus neutralization tests. The levels of neutralizing antibodies were evaluated for an ancestral D614G strain and a recent circulating SARS-CoV-2 variant of concern (Omicron BA.4). The results revealed that the vaccine induced a strong antibody response in mink by reaching antibody titer levels of up to 1:12800 in the ELISA. Moreover, high levels of neutralizing antibodies were obtained, and despite the great level of genetic differences between the ancestral and Omicron BA.4 strains, the vaccinated mink showed high levels of cross-reacting neutralizing antibodies. Interestingly, the antibody levels towards SARS-CoV-2 in the Canadian vaccinated mink were significantly higher than observed in recently SARS-CoV-2 infected Danish mink and equal to anamnestic responses following re-infection. In conclusion, the vaccine used in the Canadian farmed mink was able to induce a strong and broad-reacting antibody response in mink, which could limit the spread of SARS-CoV-2 in farmed mink and thereby reduce the risk of mink serving as a SARS-CoV-2 reservoir for human infections.

Are patients with eosinophilic esophagitis treated at an academic hospital comparable to a patient from a population-based cohort? Not in Denmark.

Background and Aim: Little are known about differences in eosinophilic esophagitis (EoE) patients in the general population compared with patients treated at academic hospitals. This might affect the generalizability of study results. The aims of the study were to compare clinical features, and complications of EoE between patients from a population-based cohort (DanEoE) and patients from an academic hospital cohort in Copenhagen (EoE-Cph). Methods: The DanEoE cohort is a population- and register-based cohort including all 236 adult EoE patients diagnosed in the North Denmark Region in 2007-2017 previously described in detail. The new EoE-Cph cohort consists of 245 consecutively referred adult patients to a dedicated EoE center in an Academic Hospital in the Danish capital in 2013-2020. Data were collected from medical registries and medical files. Results: Patients in the academic cohort were at symptom debut 12 (SD 16) years younger (P = 0.001). At the time of diagnosis they were 5.4 (SD 15) years younger (P < 0.001). Where Gastro-esophageal reflux disease (GORD) was present in one-third of the population-based cohort, this was only observed in 14% of the EoE-Cph group (P < 0.05). Food bolus obstruction before diagnosis was 24% less common in the EoE-Cph patients (P < 0.001). Conclusion: Results indicated that EoE patients referred to a Danish EoE center is a selected subgroup with disease debut at a younger age, less comorbid GORD, and rarely food bolus obstruction before diagnosis. This suggests that study results from academic hospitals might not have generalizability to the average EoE patient in a population.

Adrenocorticotropic hormone secretion in response to anticipatory stress and venepuncture: The role of menstrual phase and oral contraceptive use.

INTRODUCTION: Oral contraceptives (OCs) are primarily known for their effects on the reproductive system, but they can also impact the hypothalamic-pituitary-adrenal (HPA) axis. The present study aimed to compare plasma adrenocorticotropic hormone (ACTH) responses to the anticipatory stress of participating in a scientific experiment and venepuncture in OC users versus naturally cycling (NC) women, with a focus on variations throughout the menstrual cycle. METHODS: We recruited 131 young women (average age 20.5) and obtained blood samples to measure plasma ACTH concentrations immediately after venepuncture and again after 15 min of group activities designed to facilitate interpersonal attachment and stress-buffering. RESULTS: ACTH levels decreased in 70% of all participants throughout the group activities. A two-way repeated measures ANOVA highlighted a significant interaction between time and OC use, indicating differential changes in ACTH levels during social interaction between OC users and NC women. Further, the post-hoc analysis revealed that a period of stress-buffering group activities significantly decreased ACTH levels in NC women during menstrual and secretory phases, but not during the proliferative phase. In contrast, OC users did not display a decrease during group activities, regardless of the phase. CONCLUSION: This study underscores the influence of OC use on stress regulation, demonstrating that OCs not only modulate reproductive functions but also impact ACTH stress reactivity. Additionally, it emphasizes the importance of considering hormonal contraceptive use and menstrual cycle phases when assessing female stress responses.

Export of defensive glucosinolates is key for their accumulation in seeds.

Plant membrane transporters controlling metabolite distribution contribute key agronomic traits1-6. To eliminate anti-nutritional factors in edible parts of crops, the mutation of importers can block the accumulation of these factors in sink tissues7. However, this often results in a substantially altered distribution pattern within the plant8-12, whereas engineering of exporters may prevent such changes in distribution. In brassicaceous oilseed crops, anti-nutritional glucosinolate defence compounds are translocated to the seeds. However, the molecular targets for export engineering of glucosinolates remain unclear. Here we identify and characterize members of the USUALLY MULTIPLE AMINO ACIDS MOVE IN AND OUT TRANSPORTER (UMAMIT) family-UMAMIT29, UMAMIT30 and UMAMIT31-in Arabidopsis thaliana as glucosinolate exporters with a uniport mechanism. Loss-of-function umamit29 umamit30 umamit31 triple mutants have a very low level of seed glucosinolates, demonstrating a key role for these transporters in translocating glucosinolates into seeds. We propose a model in which the UMAMIT uniporters facilitate glucosinolate efflux from biosynthetic cells along the electrochemical gradient into the apoplast, where the high-affinity H+-coupled glucosinolate importers GLUCOSINOLATE TRANSPORTERS (GTRs) load them into the phloem for translocation to the seeds. Our findings validate the theory that two differently energized transporter types are required for cellular nutrient homeostasis13. The UMAMIT exporters are new molecular targets to improve nutritional value of seeds of brassicaceous oilseed crops without altering the distribution of the defence compounds in the whole plant.

No gender differences in patients with eosinophilic oesophagitis.

INTRODUCTION: Gender difference in the incidence of eosinophilic oesophagitis (EoE) is well-known as more men than women are affected. However, knowledge of gender differences is lacking for most other aspects of EoE. In this population-based adult EoE cohort, the aim was to study if gender differences exist with respect to 1) clinical phenotype, 2) treatment response and 3) complications. METHODS: This was a retrospective, registry-based DanEoE cohort study of 236 adult patients with EoE (178 adult men and 58 adult women) diagnosed in 2007-2017 in the North Denmark Region. Medical registries were searched for patient records and pathology reports. RESULTS: No statistically or clinically significant differences were recorded in the phenotype regarding symptoms reported, macroscopic or histological findings at diagnosis (all p > 0.3). A comparable number of men and women were followed up symptomatically and histologically (all p > 0.3). More men than women reported "no symptoms" on proton pump inhibitor (56% men versus 39% women, p = 0.04), although the histological response was similar between genders (p = 0.4). The proportions of food bolus obstructions and dilations were comparable (all p > 0.4). CONCLUSION: This study found very few gender differences. Results suggest that men and women with EoE may receive the same treatment. FUNDING: none. TRIAL REGISTRATION: not relevant.

H2 O2 feeding enables LPMO-assisted cellulose saccharification during simultaneous fermentative production of lactic acid.

Simultaneous saccharification and fermentation (SSF) is a well-known strategy for valorization of lignocellulosic biomass. Because the fermentation process typically is anaerobic, oxidative enzymes found in modern commercial cellulase cocktails, such as lytic polysaccharide monooxygenases (LPMOs), may be inhibited, limiting the overall efficiency of the enzymatic saccharification. Recent discoveries, however, have shown that LPMOs are active under anoxic conditions if they are provided with H2 O2 at low concentrations. In this study, we build on this concept and investigate the potential of using externally added H2 O2 to sustain oxidative cellulose depolymerization by LPMOs during an SSF process for lactic acid production. The results of bioreactor experiments with 100 g/L cellulose clearly show that continuous addition of small amounts of H2 O2 (at a rate of 80 µM/h) during SSF enables LPMO activity and improves lactic acid production. While further process optimization is needed, the present proof-of-concept results show that modern LPMO-containing cellulase cocktails such as Cellic CTec2 can be used in SSF setups, without sacrificing the LPMO activity in these cocktails.

Effect of nitrification inhibitor (DMPP) on nitrous oxide emissions from agricultural fields: Automated and manual measurements.

Nitrogen fertilisation contributes significantly to the atmospheric increase of nitrous oxide (N2O). Application of nitrification inhibitors (NIs) is a promising strategy to mitigate N2O emissions and improve N-use efficiency in agricultural systems. This study investigated the effect of NI, 3,4-dimethylpyrazol phosphate (DMPP) on N2O mitigation from spring barley and spring oilseed rape. Manual and automatic chamber methodologies were used to capture spatial and temporal variability in N2O emissions. In a second experiment, we study the effect of N fertiliser levels without NI (0 %, 50 %, 100 %, 150 % and 200 % of recommended amount of N fertiliser), as well as 100 % of N with NI on N2O emissions in spring barley. The automated chamber measurements showed dynamics of N2O changes throughout the season, including positive and negative peaks that were unobservable with manual chambers due to low temporal resolution. Although not significant, application of NI tended to reduce N2O emissions. The reduction was on average 16 % in spring barley and 58 % in spring oilseed rape in manual chamber measurements. However, N2O reduction was 108 % in continuous automatic chamber measurements in spring barley. The N2O EFs for the growing season were very low (0.025 % to 0.148 %), with a greater reduction in EF in spring oilseed rape (76 %) than in spring barley (32 %) with NI application. A positive correlation (R = 80 %) was observed between N fertiliser levels and N2O emissions. Crop yield and crop N uptake were not significantly affected by the use of NI. This study highlighted that NI can reduce N2O emissions, but the reduction effects are plot, crop and microclimate specific. Long-term experiments with continuous plot-scale measurements are needed to capture and optimise N2O mitigation effect of NIs across wide variability in soils and microclimates in agroecosystems.

Aptamer-Targeted Drug Delivery for Staphylococcus aureus Biofilm.

Treatment of Staphylococcus aureus biofilm infections using conventional antibiotic therapy is challenging as only doses that are sublethal to the biofilm can be administered safely to patients. A potential solution to this challenge is targeted drug delivery. In this study, we tailored an aptamer-targeted liposomal drug delivery system for accumulation and delivery of antibiotics locally in S. aureus biofilm. In our search for a suitable targeting ligand, we identified six DNA aptamers that bound to S. aureus cells in biofilms, and we demonstrated that one of these aptamers could facilitate accumulation of liposomes around S. aureus cells inside the biofilm. Aptamer-targeted liposomes encapsulating a combination of vancomycin and rifampicin were able to eradicate S. aureus biofilm upon 24 h of treatment in vitro. Our results point to that aptamer-targeted drug delivery of antibiotics is a potential new strategy for treatment of S. aureus biofilm infections.

Growth-enhanced salmon modify stream ecosystem functioning.

Use of fast-growing domesticated and/or genetically modified strains of fish is becoming increasingly common in aquaculture, increasing the likelihood of deliberate or accidental introductions into the wild. To date, their ecological impacts on ecosystems remain to be quantified. Here, using a controlled phenotype manipulation by implanting growth hormone in juvenile Atlantic salmon (Salmo salar), we found that growth-enhanced fish display changes in several phenotypic traits known to be important for ecosystem functioning, such as habitat use, morphology and excretion rate. Furthermore, these phenotypic changes were associated with significant impacts on the invertebrate community and key stream ecosystem functions such as primary production and leaf-litter decomposition. These findings provide novel evidence that introductions of growth-enhanced fish into the wild can affect the functioning of natural ecosystems and represent a form of intraspecific invasion. Consequently, environmental impact assessments of growth-enhanced organisms need to explicitly consider ecosystem-level effects.

Modelling the downstream longitudinal effects of frequent hydropeaking on the spawning potential and stranding susceptibility of salmonids.

Hydropower plant operating conditions are expected to change to be more in tandem with intermittent power production so as to meet the requirements of the Paris Agreement, which in turn may negatively impact ecological conditions downstream of the hydropower plants. The current study investigates how highly flexible hydropower operating conditions may impact several salmonid species (European grayling, Atlantic salmon and brown trout) in the River Umeälven, a major river in northern Sweden; specifically, how changes in hydropeaking frequency may affect the area of the downstream watercourse that is hydraulically suitable for spawning (potential spawning area) and how changes in spill gate closing time may affect the propensity to stranding. River hydrodynamics were modeled using the open-source solver Delft3D, with a range of hydropeaking frequencies (from 10 to 60 starts and stops per day) and a range of spill gate closing times from (1-30 min). Increasing the hydropeaking frequency caused a reduction in potential spawning area, but also a reduction in dewatering of potential spawning area at low flows. Increasing spill gate closing time caused a decrease in propensity to stranding. Effects were dependent on both species and life-stage, and declined longitudinally with distance downstream from the spillway outlet. The modelling approach used here provides an effective method for predicting likely outcomes of flexible hydropower operating conditions, taking into account fish species and life-stages present and watercourse characteristics.

Translational challenges of remote ischemic conditioning in ischemic stroke - a systematic review.

Remote ischemic conditioning (RIC) has well-established cardioprotective effects in preclinical studies and promising results in preclinical stroke research. Effective translation from preclinical studies to clinical trials has yet to be accomplished, perhaps because of the use of multiple applications of RIC (e.g., pre-, per-, or post-conditioning) in preclinical studies by both invasive and non-invasive protocols, some of which not clinically applicable. Our systematic review conformed to PRISMA guidelines and addressed differences in clinically relevant RIC applications and outcomes between preclinical and clinical studies. We retrieved a total of 30 studies (8 human; 22 animal) that met the inclusion criteria of testing clinically relevant procedures; namely, non-invasive and per- or post-conditioning protocols. Per-conditioning was applied in 6 animal and 3 human studies, post-conditioning was applied in 16 animal and 5 human studies, and both conditioning methods were applied in 2 animal studies. Application of RIC varied between human and animal studies regarding initiation, duration, repetition, and number of limbs included. Study designs did not systematically apply blinding, randomization, or placebo controls. On only a few occasions did preclinical studies include animals with clinically relevant comorbidities. Clinical trials were challenged by not completing the intended number of RIC cycles or addressing this deficit in the data analysis. Consistency and transferability of methods used for positive animal studies and subsequent human studies are essential for the optimal translation of results. Consensus on preclinical and clinical RIC procedures should be reached for a full understanding of the possible beneficial effects of RIC treatment in stroke.

Characteristics of existing healthcare workforce education in spiritual care related to childbirth: A systematic review identifying only two studies.

BACKGROUND: It has been argued that the beginning of life is one of the most significant, universally-shared life events, impacting parental health biologically, sociologically, psychologically and spiritually. In maternity care settings, only a few educational initiatives exist focusing on increasing competencies in spiritual care. OBJECTIVE: To explore the characteristics of content in existing under- and post-graduate education of healthcare professionals in spiritual care in the field of maternity care. METHODS: We conducted an integrative review, searching seven databases for studies describing the content of existing education in spiritual care in maternity care settings. RESULTS: From 235 studies assessed eligible and full text screened, only two were included, originating from the same project. The majority of existing studies about spiritual care focus on the perspective of women related to loss, sickness or bereavement, whereas research related to the field of maternity care is sparser. Furthermore, the perspective of the professional seems overlooked. CONCLUSION: There is a lack of research exploring the content and structure of educational initiatives related to spiritual care in maternity care. In order to strengthen spiritual care competencies in maternity care, for both women/partners and professionals, future research should investigate how education is planned and evaluated.

How Giving Birth Makes Sense: A Questionnaire Study on Existential Meaning-Making Among Mothers Giving Birth Preterm or at Term.

BACKGROUND: Women's reflections on existential meaning-making in relation to giving birth may seem indistinct in maternity services and have not been thoroughly explored in secular contexts. However, research suggests that childbirth accentuates spiritual and existential considerations and needs even in secular contexts highlighting the importance of care for such needs in maternity care practices. The objectives of this study were two-fold: Firstly, to explore how first-time mothers, living in a secular context, experience their first birth in relation to existential meaning-making. Secondly, to describe the relationship between existential meaning-making reflections and gestational week at birth. METHODS: A nationwide cross-sectional study in Denmark based on the questionnaire "Faith, existence and motherhood" was conducted in 2011. Eight core items related to birth experience informed this study. The cohort was sampled from the Danish Medical Birth Registry and consisted of 913 mothers having given birth 6-18 months previously. Twenty-eight per cent had given birth preterm (PT) and 72% had given birth at full-term (FT). A total of 517 mothers responded. RESULTS: In relation to the birth of their first child, both FT and PT mothers answered, that they had existential meaning-making reflections. The consent to the 8 items ranged from 17 to 73% among FT mothers and from 19 to 58% among PT mothers. Mothers who gave birth preterm mainly identified the negative aspects of birth, whereas mothers, who gave birth at full-term, to a higher degree identified positive aspects. CONCLUSIONS: Findings suggest that not only traumatic birth events accentuate existential reflections, but that even normal childbirth to most mothers is an existential event. However, the quality of existential reflections differs when comparing normal and traumatic birth. The study points towards change in education and organization of maternity care to better care for existential needs and reflections specific to every new mother and birthing woman.

Simple Quantitative Sensory Testing Reveals Paradoxical Co-existence of Hypoesthesia and Hyperalgesia in Diabetes.

Background: Diabetic neuropathy is characterized by the paradoxical co-existence of hypo- and hyperalgesia to sensory stimuli. The literature shows consistently sensory differences between healthy and participants with diabetes. We hypothesized that due to differences in pathophysiology, advanced quantitative sensory testing (QST) might reveal sensory discrepancies between type 1 (T1D) and type 2 diabetes (T2D). Furthermore, we investigated whether vibration detection thresholds (VDT) were associated with sensory response. Method: Fifty-six adults with T1D [43 years (28-58)], 99 adults with T2D [65 years (57-71)], and 122 healthy individuals [51 years (34-64)] were included. VDT, pressure pain detection thresholds (pPDT) and tolerance (pPTT), tonic cold pain (hand-immersion in iced water), and central pain mechanisms (temporal summation and conditioned pain modulation) were tested and compared between T1D and T2D. VDT was categorized into normal (< 18 V), intermediary (18-25 V), or high (> 25 V). Results: In comparison to healthy, analysis adjusted for age, BMI, and gender revealed hypoalgesia to tibial (pPDT): p = 0.01, hyperalgesia to tonic cold pain: p < 0.01, and diminished temporal summation (arm: p < 0.01; abdomen: p < 0.01). In comparison to participants with T2D, participants with T1D were hypoalgesic to tibial pPDT: p < 0.01 and pPTT: p < 0.01, and lower VDT: p = 0.02. VDT was not associated with QST responses. Conclusion: Participants with T1D were more hypoalgesic to bone pPDT and pPTT independent of lower VDT, indicating neuronal health toward normalization. Improved understanding of differentiated sensory profiles in T1D and T2D may identify improved clinical endpoints in future trials.

The vasopressor action of angiotensin II (ANG II) in ball pythons (Python regius).

Angiotensin II (ANG II) is part of the renin-angiotensin system (RAS) in vertebrates and exert vasoconstriction in all species studied. The present study examines the vasopressor effect of ANG II in the ball python (Python regius), and examines whether ANG II exert its effect through direct angiotensin receptors or through an activation of α-adrenergic receptors. The studies were conducted in snakes with chronic arterial catheters that had recovered from anesthesia. In addition to demonstrating a clear and pronounced dose-dependent rise in arterial blood pressure upon repeated injections of boluses with ANG II (0.001-1 µg/kg), we demonstrate that the pressor response persisted following α-adrenergic blockade using the α-adrenergic antagonist phentolamine (2.5 mg/kg). Unfortunately, it proved impossible to block the ANG receptors using losartan (1, 3 or even 10 mg/kg). The pressor response to ANG II was associated with a significant rise in heart rate at the higher dosages, pointing to a resetting of the barostatic mechanism for heart rate regulation. The responses were similar in fasting and digesting pythons despite the expected rise in baseline values for blood pressure and heart rate of the digesting snakes.

Enzymatic processing of lignocellulosic biomass: principles, recent advances and perspectives.

Efficient saccharification of lignocellulosic biomass requires concerted development of a pretreatment method, an enzyme cocktail and an enzymatic process, all of which are adapted to the feedstock. Recent years have shown great progress in most aspects of the overall process. In particular, increased insights into the contributions of a wide variety of cellulolytic and hemicellulolytic enzymes have improved the enzymatic processing step and brought down costs. Here, we review major pretreatment technologies and different enzyme process setups and present an in-depth discussion of the various enzyme types that are currently in use. We pay ample attention to the role of the recently discovered lytic polysaccharide monooxygenases (LPMOs), which have led to renewed interest in the role of redox enzyme systems in lignocellulose processing. Better understanding of the interplay between the various enzyme types, as they may occur in a commercial enzyme cocktail, is likely key to further process improvements.

Crystal Structures of Human C4.4A Reveal the Unique Association of Ly6/uPAR/α-neurotoxin Domain.

Ly6/uPAR/α-neurotoxin domain (LU-domain) is characterized by the presence of 4-5 disulfide bonds and three flexible loops that extend from a core stacked by several conversed disulfide bonds (thus also named three-fingered protein domain). This highly structurally stable protein domain is typically a protein-binder at extracellular space. Most LU proteins contain only single LU-domain as represented by Ly6 proteins in immunology and α-neurotoxins in snake venom. For Ly6 proteins, many are expressed in specific cell lineages and in differentiation stages, and are used as markers. In this study, we report the crystal structures of the two LU-domains of human C4.4A alone and its complex with a Fab fragment of a monoclonal anti-C4.4A antibody. Interestingly, both structures showed that C4.4A forms a very compact globule with two LU-domain packed face to face. This is in contrast to the flexible nature of most LU-domain-containing proteins in mammals. The Fab combining site of C4.4A involves both LU-domains, and appears to be the binding site for AGR2, a reported ligand of C4.4A. This work reports the first structure that contain two LU-domains and provides insights on how LU-domains fold into a compact protein and interacts with ligands.