Nitrous oxide emissions from soybean in response to drained and undrained soils and previous corn nitrogen management.

While corn (Zea mays L.)-soybean (Glycine max. Merr. L) is a predominant rotation system in the US Midwest the residual effect of nitrogen (N) fertilization to corn on the following year's soybean and N2O emissions under different soil drainage conditions has not been studied. Our objective was to quantify agronomic parameters and season-long N2O emissions from soybean as affected by N management (0-N and optimum N rate of 135 kg N ha-1 as single or split application) during the previous corn crop under drained and undrained systems. Urea was applied to corn, and residual N effects were measured on soybean the following year in a poorly drained soil with and without subsurface tile drainage. Drainage reduced N2O emissions in one of three growing seasons but had no effect on soybean yield or N removal in grain. Nitrogen management in the previous corn crop had no effect on soybean grain yield, N removal, or N2O emissions during the soybean phase. Even though soybean symbiotically fixes N and removes more N in grain than corn, N2O emissions were more than two times greater during the corn phase (mean = 1.83 kg N ha-1) due to N fertilization than during the soybean phase (mean = 0.80 kg N ha-1). Also, N2O emissions in the corn years were increased possibly due to decomposition of the previous year's soybean crop residue compared to corn residue decomposition in the soybean years. Tile drainage, especially where wet soil conditions are prevalent, is a viable option to mitigate agricultural N2O emissions.

Dataset on target chemical and bioassay analysis-Exploring contaminants of emerging concern in a low mountain river of central Germany.

Chemical pollution of the aquatic environment is nowadays characterised by increasing levels of anthropogenic organic compounds at low concentrations and is recognised as one of the main drivers of the deteriorated ecological state of European waterbodies. To improve the understanding of the impact of chemical pollution in surface waters, a combined approach of chemical and bioanalytical testing is considered necessary for effective ecologically oriented water management. For this dataset, six 25-L water samples were collected at six sampling sites along the Holtemme River in Central Germany using large-volume solid phase extraction. All samples were analysed by targeted high-resolution liquid chromatography-mass spectrometry (LC-MS) and a selected bioanalytical test battery using effect-based methods. These methods included cytotoxicity assessment, several mechanism-specific CALUXⓇ tests to identify endocrine and oxidative stress-related effects and the fish embryo acute toxicity test to investigate (sub)lethal effects in the model species Danio rerio. This approach provided a dataset that offers a longitudinal characterisation of the chemical pollution and ecotoxicological impacts. The combination of chemical analysis and effect-based analysis is valuable for future studies as it will help researchers, risk assessors and authorities to identify hot spots of chemical pollution, monitor environmental quality standards and recommend mitigation strategies.

Development of a Tag/Catcher-mediated capsid virus-like particle vaccine presenting the conserved Neisseria gonorrhoeae SliC antigen that blocks human lysozyme.

Virus-like particles (VLPs) are promising nanotools for the development of subunit vaccines due to high immunogenicity and safety. Herein, we explored the versatile and effective Tag/Catcher-AP205 capsid VLP (cVLP) vaccine platform to address the urgent need for the development of an effective and safe vaccine against gonorrhea. The benefits of this clinically validated cVLP platform include its ability to facilitate unidirectional, high-density display of complex/full-length antigens through an effective split-protein Tag/Catcher conjugation system. To assess this modular approach for making cVLP vaccines, we used a conserved surface lipoprotein, SliC, that contributes to the Neisseria gonorrhoeae defense against human lysozyme, as a model antigen. This protein was genetically fused at the N- or C-terminus to the small peptide Tag enabling their conjugation to AP205 cVLP, displaying the complementary Catcher. We determined that SliC with the N-terminal SpyTag, N-SliC, retained lysozyme-blocking activity and could be displayed at high density on cVLPs without causing aggregation. In mice, the N-SliC-VLP vaccines, adjuvanted with AddaVax or CpG, induced significantly higher antibody titers compared to controls. In contrast, similar vaccine formulations containing monomeric SliC were non-immunogenic. Accordingly, sera from N-SliC-VLP-immunized mice also had significantly higher human complement-dependent serum bactericidal activity. Furthermore, the N-SliC-VLP vaccines administered subcutaneously with an intranasal boost elicited systemic and vaginal IgG and IgA, whereas subcutaneous delivery alone failed to induce vaginal IgA. The N-SliC-VLP with CpG (10 µg/dose) induced the most significant increase in total serum IgG and IgG3 titers, vaginal IgG and IgA, and bactericidal antibodies.

Prevalence, causes, and clinical course of patients with surgical reintervention due to complications during cesarean section.

OBJECTIVE: The objective of the study is to identify the prevalence, causes, and clinical evolution of patients with surgical reintervention due to complications during cesarean section. MATERIALS AND METHODS: The file of the Toco-Surgical Unit of the Gynecological Obstetrics Hospital No. 3 of the National Medical Center "La Raza" Mexican Institute of Social Security was reviewed to know the total number of patients undergoing cesarean section from January to December 2019 and cases with reintervention due to complications during cesarean section were selected. Their general data, the cause of reintervention, stay in the intensive care unit (ICU), hospital stay, and mortality were studied. The data were analyzed with descriptive statistics using the statistical program SPSS version 20. RESULTS: It was found that 3371 patients underwent cesarean section, of which 1.60% (54 cases) underwent reoperation for the following reasons: Unpacking 27.79%, obstetric hemorrhage 20.37%, bleeding due to uterine atony 20.37%, hysterotomy commissure hematoma 18.52%, uterine infiltration 3.70%, vascular injury 3.70%, bladder injury 3.70%, and colonic injury 1.85%. The ICU stay was 3.79 ± 2.03 days, hospital stay was 13.67 ± 11.16 days, and mortality was 1.85%. CONCLUSION: The prevalence of reintervention was reduced, bleeding was the main cause, and the clinical evolution was satisfactory with low mortality.

OBJETIVO: Identificar la prevalencia, causas y evolución clínica de las pacientes con reintervención quirúrgica por complicaciones durante la cesárea. MATERIAL Y MÉTODOS: Se revisó el archivo de la Unidad de Toco-Quirúrgica del Hospital Ginecobstetricia No. 3 del Centro Médico Nacional "La Raza" Instituto Mexicano del Seguro Social para conocer el total de pacientes sometidas a cesárea desde enero hasta diciembre de 2019 y se seleccionaron los casos con reintervención por complicaciones durante la cesárea. Se estudiaron sus datos generales, la causa de reintervención, estancia en la Unidad de Cuidados Intensivos (UCI), estancia en hospital y la mortalidad. Los datos se analizaron con estadística descriptiva utilizando el programa estadístico SPSS versión 20. RESULTADOS: Se encontró que 3371 pacientes fueron sometidas a cesárea de las cuales 1.60% (54 casos) se reintervinieron por las siguientes causas: desempaquetamiento 27.79%, hemorragia obstétrica 20.37%, sangrado por atonía uterina 20.37%, hematoma de la comisura de histerotomía 18.52%, infiltración uterina 3.70%, lesión vascular 3.70%, lesión vesical 3.70% y lesión colónica 1.85%. La estancia en UCI fue 3.79 ± 2.03 días, estancia en hospital 13.67 ± 11.16 días y mortalidad 1.85%. CONCLUSIÓN: La prevalencia de reintervención fue reducida, el sangrado fue la principal causa y la evolución clínica resultó satisfactoria con baja mortalidad.

Immature stages of the limnephilid caddisfly Verger lutzi (Navás 1918) (Trichoptera: Limnephilidae): description and larval life-history traits in seasonal forested wetlands of Northwestern Patagonia.

The genus Verger Navás 1918 (Trichoptera: Limnephilidae), is a Neotropical genus exclusive to the Andean region, with most of its species distributed from Tierra del Fuego to north central Chile and Argentina. Currently, 20 species of the genus have been described from adults and only six larvae have been associated and diagnosed. The ecology of the species is completely unknown. In this work, we describe and illustrate the immature stages of Verger lutzi (Navás 1918) and study its breeding phenology, larval growth and adult emergence. The main characters that enable the separation of this species are the coloration the body, shape of the anterior ventral apotome, shape and distribution of metanotal setal areas, distribution of tracheal gills and number of accessory teeth in the anal prolegs. Reproduction occurs during the summer and females lay their eggs on the wetland basin formed by humid leaf litter and organic debris, covered by herbaceous vegetation. Larvae hatch after flooding and overwinter in the wetland. Larvae develop fast during spring and emergence begins in December before the wetland dries up. Description of the immature stages is helpful for use of macroinvertebrates biomonitoring, ecological studies and understanding their ecosystem services in seasonal-lentic habitats.

Evidence for operative treatment of talar osteochondral lesions: a systematic review.

Purpose: Operative treatment of talar osteochondral lesions is challenging with various treatment options. The aims were (i) to compare patient populations between the different treatment options in terms of demographic data and lesion size and (ii) to correlate the outcome with demographic parameters and preoperative scores. Methods: A systemic review was conducted according to the PRISMA guidelines. The electronic databases Pubmed (MEDLINE) and Embase were screened for reports with the following inclusion criteria: minimum 2-year follow-up after operative treatment of a talar osteochondral lesion in at least ten adult patients and published between 2000 and 2020. Results: Forty-five papers were included. Small lesions were treated using BMS, while large lesions with ACI. There was no difference in age between the treatment groups. There was a correlation between preoperative American Orthopaedic Foot and Ankle Society (AOFAS) score and change in AOFAS score (R = -0.849, P < 0.001) as well as AOFAS score at follow-up (R = 0.421, P = 0.008). Preoperative size of the cartilage lesion correlates with preoperative AOFAS scores (R= -0.634, P = 0.001) and with change in AOFAS score (R = 0.656, P < 0.001) but not with AOFAS score at follow-up. Due to the heterogeneity of the studies, a comparison of the outcome between the different operative techniques was not possible. Conclusion: Patient groups with bigger lesions and inferior preoperative scores did improve the most after surgery. Level of evidence: IV.

Leaf surface traits contributing to wettability, water interception and uptake of above-ground water sources in shrubs of Patagonian arid ecosystems.

BACKGROUND AND AIMS: The ecohydrological significance of leaf wetting due to atmospheric water in arid and semiarid ecosystems is not well understood. In these environments, the inputs of precipitation or dew formation resulting in leaf wetting have positive effects on plant functioning. However, its impact on plant water relations may depend on the degree of leaf surface wettability. In this study we evaluated leaf wettability and other leaf traits and its effects on foliar water uptake and canopy interception in plant species of a Patagonian steppe. We also studied how leaf traits affecting wettability vary seasonally from growing to dry season. METHODS: Contact angle of a water droplet with the leaf surface, water adhesion, droplet retention angle, stomatal density, cuticular conductance, canopy interception and maximum foliar water uptake were determined in six dominant shrub species. KEY RESULTS: All species increased leaf wettability during the dry season and most species were considered highly wettable. The leaf surface had very high capacity to store and retain water. We found a negative correlation between foliar water uptake and leaf hydrophilia. CONCLUSIONS: Despite the diversity of life forms, including cushion shrubs and tall shrubs, as well as phenological variability, all species converged in similar seasonal changes in leaf traits that favour wettability. Intercepted water by crowns and the extremely high capacity of retention of droplets on leaf surfaces can have a significant impact on eco-hydrological process in water limited ecosystems where most of water sources during the growing and the dry season may be small rainfall events or dew, which do not always increase soil water availability.

Antifungal susceptibility of Malassezia pachydermatis isolated from the external auditive conduct from dogs, in central Chile.

Background: External otitis is common in dogs, and one of the main agents involved is Malassezia pachydermatis, a yeast belonging to the otic microbiota. Empirical treatment can fail; therefore, it is essential to know the antifungal susceptibility profile to prescribe appropriate treatment, a fact scarcely reported in Chile. Aim: This study aimed to determine the antifungal sensitivity of M. pachydermatis isolated from the external auditory canal of dogs in central Chile. Methods: Ear swabs from 30 dogs (15 healthy and 15 with external otitis) were used. Samples were subjected to cytology and fungal culture. The antifungal susceptibility was determined according to clinical and laboratory standards institute (CLSI) document M44A-2 using the disk diffusion test from amphotericin B, Caspofungin, fluconazole, nystatin, clotrimazole, and voriconazole were used. Results: The prevalence of M. pachydermatis was 66.7% from 8 healthy dogs and 12 with otitis. While fungal culture was not associated with the patient's clinical condition (p = 0.24), the yeast count by cytology was significantly higher in dogs with otitis (p = 0.003). The strains were sensitive to all antifungals except for Caspofungin, where 55% of the strains were resistant. Conclusion: Malassezia pachydermatis is isolated more frequently in dogs with otitis, and the ear cytological examination is useful to differentiate colonized dogs versus dogs with otitis. In addition, most antifungals in vitro are active against this yeast, except Caspofungin, an antifungal used in human medicine. This situation should be further monitored in epidemiological programs to evaluate the possible impact on Chilean public health.

Evaluation of Radiographic Features Including Metatarsus Primus Elevatus in Hallux Rigidus.

The etiology of hallux rigidus remains a controversial issue in foot and ankle surgery, i.e., the relationship between metatarsus primus elevatus (MPE) and hallux rigidus. The purpose of this study was to evaluate several radiographic parameters including first metatarsal elevation in patients with hallux rigidus compared to a matched control group. A retrospective case control study was performed including 50 feet, 25 feet with and 25 feet without hallux rigidus. In the patients with hallux rigidus, the first metatarsal was more elevated than in the control group (8.3 ± 1.7 mm vs 3.0 ± 2.0 mm, p < .001) and in 60% of patients with hallux rigidus MPE was diagnosed, compared to zero patients in the control group (p < .001). The lateral 1 to 2 intermetatarsal angle was higher in patients with hallux rigidus (3.6 ± 2.5 vs -0.7 ± 2.8; p < .001). The first metatarsal declination angle was not different between the 2 groups. Intraclass correlation coefficient between 2 observers for measuring the first metatarsal elevation was 0.929 (p < .001). In the current study, increased elevation of the first metatarsal, a higher incidence of MPE and increased lateral 1 to 2 intermetatarsal angle were found in patients with hallux rigidus compared to the control group. These findings support the theory of an association between MPE and hallux rigidus. Further high reliability of first metatarsal elevation measurement was found in our study.

A plea for the integration of Green Toxicology in sustainable bioeconomy strategies - Biosurfactants and microgel-based pesticide release systems as examples.

A key aspect of the transformation of the economic sector towards a sustainable bioeconomy is the development of environmentally friendly alternatives for hitherto used chemicals, which have negative impacts on environmental health. However, the implementation of an ecotoxicological hazard assessment at early steps of product development to elaborate the most promising candidates of lowest harm is scarce in industry practice. The present article introduces the interdisciplinary proof-of-concept project GreenToxiConomy, which shows the successful application of a Green Toxicology strategy for biosurfactants and a novel microgel-based pesticide release system. Both groups are promising candidates for industrial and agricultural applications and the ecotoxicological characterization is yet missing important information. An iterative substance- and application-oriented bioassay battery for acute and mechanism-specific toxicity within aquatic and terrestrial model species is introduced for both potentially hazardous materials getting into contact with humans and ending up in the environment. By applying in silico QSAR-based models on genotoxicity, endocrine disruption, skin sensitization and acute toxicity to algae, daphnids and fish, individual biosurfactants resulted in deviating toxicity, suggesting a pre-ranking of the compounds. Experimental toxicity assessment will further complement the predicted toxicity to elaborate the most promising candidates in an efficient pre-screening of new substances.

Ankle Joint Pressure in Supination-External Rotation Injuries: A Biomechanical Study in an Unrestrained Cadaver Model.

BACKGROUND: Previous biomechanical studies simulating supination-external rotation (SER) IV injuries revealed different alterations in contact area and peak pressure. We investigated joint reaction forces and radiographic parameters in an unrestrained, more physiological setup. METHODS: Twelve lower leg specimens were destabilized stepwise by osteotomy of the fibula (SER II) and transection of the superficial (SER IVa) and the deep deltoid ligament (SER IVb) according to the Lauge-Hansen classification. Sensors in the ankle joint recorded tibio-talar pressure changes with axial loading at 700 N in neutral position, 10° of dorsiflexion, and 20° of plantarflexion. Radiographs were taken for each step. RESULTS: Three of 12 specimen collapsed during SER IVb. In the neutral position, the peak pressure and contact area changed insignificantly from 2.6 ± 0.5 mPa (baseline) to 3.0 ± 1.4 mPa (SER IVb) (P = .35) and from 810 ± 42 mm2 to 735 ± 27 mm2 (P = .08), respectively. The corresponding medial clear space (MCS) increased significantly from 2.5 ± 0.4 mm (baseline) to 3.9 ± 1.1 mm (SER IVb) (P = .028).The position of the ankle joint had a decisive effect on contact area (P = .00), center of force (P = .00) and MCS (P = .01). CONCLUSION: Simulated SER IVb injuries demonstrated radiological, but no biomechanical changes. This should be considered for surgical decision making based on MCS width on weightbearing radiographs. LEVELS OF EVIDENCE: Not applicable. Biomechanical study.

Arthroscopically Assisted Eaton-Littler Trapeziometacarpal Ligamentoplasty.

Background Eaton-Littler ligamentoplasty has proven its efficacy in the treatment of trapeziometacarpal (TMC) instability. Description of Technique In this article, we describe the arthroscopically assisted Eaton-Littler ligamentoplasty through two clinical cases. Patients and Methods Arthroscopy is used to accurately place the metacarpal bone tunnel and pass the slip of flexor carpi radialis tendon in the latter. This procedure avoids soft-tissue dissection and could improve the outcome of this well-known surgery. Results The two clinical cases showed good short-term outcome with a decrease of pain and good function. Conclusions Arthroscopy to assist Eaton-Littler TMC ligamentoplasty is technically feasible, allowing a less invasive surgery. A larger prospective controlled study with a longer term outcome is mandatory to assess the real advantages of this modified procedure.

The Utilization and Benefits of Telehealth Services by Health Care Professionals Managing Breast Cancer Patients during the COVID-19 Pandemic.

Telehealth is the delivery of many health care services and technologies to individuals at different geographical areas and is categorized as asynchronously or synchronously. The coronavirus disease 2019 (COVID-19) pandemic has caused major disruptions in health care delivery to breast cancer (BCa) patients and there is increasing demand for telehealth services. Globally, telehealth has become an essential means of communication between patient and health care provider. The application of telehealth to the treatment of BCa patients is evolving and increasingly research has demonstrated its feasibility and effectiveness in improving clinical, psychological and social outcomes. Two areas of telehealth that have significantly grown in the past decade and particularly since the beginning of the COVID-19 pandemic are telerehabilitation and teleoncology. These two technological systems provide opportunities at every stage of the cancer care continuum for BCa patients. We conducted a literature review that examined the use of telehealth services via its various modes of delivery among BCa patients particularly in areas of screening, diagnosis, treatment modalities, as well as satisfaction among patients and health care professionals. The advantages of telehealth models of service and delivery challenges to patients in remote areas are discussed.

Low-temperature acclimation and legacy effects of summer water deficits in olive freezing resistance.

Low temperatures and drought are the main environmental factors affecting plant growth and productivity across most of the terrestrial biomes. The objective of this study was to analyze the effects of water deficits before the onset of low temperatures in winter to enhance freezing resistance in olive trees. The study was carried out near the coast of Chubut, Argentina. Plants of five olive cultivars were grown outdoor in pots and exposed to different water deficit treatments. We assessed leaf water relations, ice nucleation temperature (INT), cell damage (LT50), plant growth and leaf nitrogen content during summer and winter in all cultivars and across water deficit treatments. Leaf INT and LT50 decreased significantly from summer to winter within each cultivar and between treatments. We observed a trade-off between resources allocation to freezing resistance and vegetative growth, such that an improvement in resistance to sub-zero temperatures was associated with lower growth in tree height. Water deficit applied during summer increased the amount of osmotically active solutes and decreased the leaf water potentials. This type of legacy effect persists during the winter after the water deficit even when treatment was removed by natural rainfalls.

A multi-biomarker study on Atlantic salmon (Salmo salar L.) affected by the emerging Red Skin Disease in the Baltic Sea.

For half a decade, the Atlantic salmon in the Baltic Sea has been facing severe health issues. Clinical signs like haemorrhage, erosions and ulcerative/necrotic skin conditions in returning adults have been reported from different Swedish rivers. These primary disease signs precede a secondary, terminal fungal infection. As initial investigations of the disease did not provide conclusive answers regarding the pathogenesis, this study was initiated to gain insight into a possible link between this so-called Red Skin Disease and anthropogenic influences. Therefore, returning salmon were caught in rivers along the Swedish coast and different tissues were sampled. The focus was put on the measurements of a battery of biomarkers as well as biochemical and haematological parameters, which were analysed using multivariate statistics. The main findings were a severe osmotic haemodilution, an immune response and an alteration of the carbohydrate metabolism in diseased fish. Furthermore, oxidative stress does not seem to be a likely factor in the pathogenesis. Concluding, certain changes in physiological parameters were shown to be indicative for the disease patterns, while others were ruled out as significant factors. Thus, this study contributes to the understanding of the Red Skin Disease and may act as a hypothesis generator for future studies.

A novel gonorrhea vaccine composed of MetQ lipoprotein formulated with CpG shortens experimental murine infection.

Bacterial surface lipoproteins are emerging as attractive vaccine candidates due to their biological importance and the feasibility of their large-scale production for vaccine manufacturing. The global prevalence of gonorrhea, resistance to antibiotics, and serious consequences to reproductive and neonatal health necessitate development of effective vaccines. Reverse vaccinology identified the surface-displayed L-methionine binding lipoprotein MetQ (NGO2139) and its homolog GNA1946 (NMB1946) as gonococcal and meningococcal vaccine candidates, respectively. Here, we assessed the suitability of MetQ for inclusion in a gonorrhea vaccine by examining MetQ conservation, its function inNeisseria gonorrhoeae (Ng) pathogenesis, and its ability to induce protective immune responses using a female murine model of lower genital tract infection. In-depth bioinformatics, phylogenetics and mapping the most prevalent Ng polymorphic amino acids to the GNA1946 crystal structure revealed remarkable MetQ conservation: ~97% Ng isolates worldwide possess a single MetQ variant. Mice immunized with rMetQ-CpG (n = 40), a vaccine containing a tag-free version of MetQ formulated with CpG, exhibited robust, antigen-specific antibody responses in serum and at the vaginal mucosae including IgA. Consistent with the activity of CpG as a Th1-stimulating adjuvant, the serum IgG1/IgG2a ratio of 0.38 suggested a Th1 bias. Combined data from two independent challenge experiments demonstrated that rMetQ-CpG immunized mice cleared infection faster than control animals (vehicle, p < 0.0001; CpG, p = 0.002) and had lower Ng burden (vehicle, p = 0.03; CpG, p < 0.0001). We conclude rMetQ-CpG induces a protective immune response that accelerates bacterial clearance from the murine lower genital tract and represents an attractive component of a gonorrhea subunit vaccine.

Medicinal Herbs Used in Traditional Management of Breast Cancer: Mechanisms of Action.

Background: Breast cancer is one of the principal causes of death among women and there is a pressing need to develop novel and effective anti-cancer agents. Natural plant products have shown promising results as anti-cancer agents. Their effectiveness is reported as decreased toxicity in usage, along with safety and less recurrent resistances compared with hormonal targeting anti-cancer agents. Methods: A literature search was conducted for all English-language literature published prior to June 2020. The search was conducted using electronic databases, including PubMed, Embase, Web of Science, and Cochrane Library. The search strategy included keywords such as breast cancer, herbs, anti-cancer biologically active components, clinical research, chemotherapy drugs amongst others. Results: The literature provides documented evidence of the chemo-preventative and chemotherapeutic properties of Ginseng, garlic (Allium sativum), Black cohosh (Actaea racemose), Tumeric (Curcuma longa), Camellia sinenis (green tea), Echinacea, Arctium (burdock), Flaxseed (Linum usitatissimum) and Black Cumin (Nigella sativa). Conclusions: The nine herbs displayed anti-cancer properties and their outcomes and mechanisms of action include inhibition of cell proliferation, angiogenesis and apoptosis as well as modulation of key intracellular pathways. However, more clinical trials and cohort human studies should be conducted to provide key evidence of their medical benefits.

Foliar water uptake in arid ecosystems: seasonal variability and ecophysiological consequences.

Foliar water uptake (FWU) has been reported for different species across several ecosystems types. However, little attention has been given to arid ecosystems, where FWU during dew formation or small rain events could ameliorate water deficits. FWU and their effects on leaf water potential (ΨLeaf) were evaluated in grasses and shrubs exploring different soil water sources in a Patagonian steppe. Also, seasonal variability in FWU and the role of cell wall elasticity in determining the effects on ΨLeaf were assessed. Eleven small rain events (< 8 mm) and 45 days with dew formation were recorded during the study period. All species exhibited FWU after experimental wetting. There was a large variability in FWU across species, from 0.04 mmol m-2 s-1 in species with deep roots to 0.75 mmol m-2 s-1 in species with shallow roots. Species-specific mean FWU rates were positively correlated with mean transpiration rates. The increase in ΨLeaf after leaf wetting varied between 0.65 MPa and 1.67 MPa across species and seasons. The effects of FWU on ΨLeaf were inversely correlated with cell wall elasticity. FWU integrated over both seasons varied between 28 mol m-2 in species with deep roots to 361 mol m-2 in species with shallow roots. Taking into account the percentage of coverage of each species, accumulated FWU represented 1.6% of the total annual transpiration of grasses and shrubs in this ecosystem. Despite this low FWU integrated over time compared to transpiration, wetting leaves surfaces can help to avoid larger water deficit during the dry season.

Bioengineering of Cytochrome P450 OleTJE: How Does Substrate Positioning Affect the Product Distributions?

The cytochromes P450 are versatile enzymes found in all forms of life. Most P450s use dioxygen on a heme center to activate substrates, but one class of P450s utilizes hydrogen peroxide instead. Within the class of P450 peroxygenases, the P450 OleTJE isozyme binds fatty acid substrates and converts them into a range of products through the α-hydroxylation, ß-hydroxylation and decarboxylation of the substrate. The latter produces hydrocarbon products and hence can be used as biofuels. The origin of these product distributions is unclear, and, as such, we decided to investigate substrate positioning in the active site and find out what the effect is on the chemoselectivity of the reaction. In this work we present a detailed computational study on the wild-type and engineered structures of P450 OleTJE using a combination of density functional theory and quantum mechanics/molecular mechanics methods. We initially explore the wild-type structure with a variety of methods and models and show that various substrate activation transition states are close in energy and hence small perturbations as through the protein may affect product distributions. We then engineered the protein by generating an in silico model of the double mutant Asn242Arg/Arg245Asn that moves the position of an active site Arg residue in the substrate-binding pocket that is known to form a salt-bridge with the substrate. The substrate activation by the iron(IV)-oxo heme cation radical species (Compound I) was again studied using quantum mechanics/molecular mechanics (QM/MM) methods. Dramatic differences in reactivity patterns, barrier heights and structure are seen, which shows the importance of correct substrate positioning in the protein and the effect of the second-coordination sphere on the selectivity and activity of enzymes.