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Purpose: Rice production in paddy soil is important for food security, and nitrogen fertilisation is important to achieve high yields. Digestate, the organic rest from biogas production can be a good fertilizer, but relatively little is known about its use in paddy soil, which is investigated here.Method: Nitrogen transformations and rice growth in soil waterlogged and at field capacity after application of digestate and digestate products were assessed in a pot experiment. Nitrogen transformations and nitrous oxide (N2O) emissions with the same digestates were also assessed in an incubation.Results: One of the tested digestates had as good fertilizer effect as urea. Nitrogen transformations went mostly as expected under given water status, but digestates were more affected by waterlogging than urea. Some instability appeared about 10 days after fertilizer application. Then nitrite levels were high, responsive microbial populations peaked and N2O emissions started. Whilst emissions after mineral fertilizer application (urea) were not affected by waterlogging, emission after application of one of the examined digestate was high at field capacity but almost zero under waterlogging. N2O emissions from all other digestate and water treatments were also low.Conclusion: Digestates have great potential as organic fertilizers in rice production, but N2O emissions under aerobic soil conditions are of concern and warrants further investigation and mitigation strategy.

Purpose: Soils are the largest store of carbon in the biosphere, but soils can also act as the key sources of greenhouse gases. Rice cultivation in paddy soil is anoxic, thus creating an optimal condition for methane production. Digestate, a potential biofertilizer, contains a microbial consortium adapted for methane production because it comes from biogas production where methane production is optimised. In this study, we explored digestate as fertiliser in paddy soil, focussing on the effects of carbon sequestration and methane emission.Method: A number of digestates and digestate products were incubated in the soils, both waterlogged and at field capacity. The effects of thermal treatment of the digestates were assessed to understand if the microbial community applied with the digestate played a role.Results: Carbon dioxide emission was 53% higher in the soil with a history of mineral fertiliser application, than in the soil with digestate application history under waterlogging, at field capacity it was 13% smaller. Methane emissions came later when the digestate was heat-treated, indicating that the microbial community in the digestate could change the timing of emissions, but not the amount. Otherwise, some digestates increased methane emissions, whilst others had little effect. The supply of available carbon appeared to be an important factor to explain differences.Conclusion: Digestate can increase carbon sequestration in paddy soil, although the interaction between waterlogging and soil history is not fully understood. The microbial community applied with the digestate can make methane emission start earlier, but it does not increase total emission.

Industrial biotechnology offers a potential ecological solution for PET recycling under relatively mild reaction conditions via enzymatic degradation, particularly using the leaf branch compost cutinase (LCC) quadruple mutant ICCG. To improve the efficient downstream processing of this biocatalyst after heterologous gene expression with a suitable production host, protein crystallization can serve as an effective purification/capture step. Enhancing protein crystallization was achieved in recent studies by introducing electrostatic (and aromatic) interactions in two homologous alcohol dehydrogenases (Lb/LkADH) and an ene reductase (NspER1-L1,5) produced with Escherichia coli. In this study, ICCG, which is difficult to crystallize, was utilized for the application of crystal contact engineering strategies, resulting in ICCG mutant L50Y (ICCGY). Previously focused on the Lys-Glu interaction for the introduction of electrostatic interactions at crystal contacts, the applicability of the engineering strategy was extended here to an Arg-Glu interaction to increase crystallizability, as shown for ICCGY T110E. Furthermore, the rationale of the engineering approach is demonstrated by introducing Lys and Glu at non-crystal contacts or sites without potential interaction partners as negative controls. These resulting mutants crystallized comparably but not superior to the wild-type protein. As demonstrated by this study, crystal contact engineering emerges as a promising approach for rationally enhancing protein crystallization. This advancement could significantly streamline biotechnological downstream processing, offering a more efficient pathway for research and industry.

Purpose Biogas residues, digestates, contain valuable nutrients and are therefore suitable as agricultural fertilizers. However, the application of fertilizers, including digestates, can enhance greenhouse gas (GHG) emissions. In this study different processes and post-treatments of digestates were analyzed with respect to triggered GHG emissions in soil. Methods In an incubation experiment, GHG emissions from two contrasting soils (chernozem and sandy soil) were compared after the application of digestate products sampled from the process chain of a food waste biogas plant: raw substrate, digestate (with and without bentonite addition), digestates after separation of liquid and solid phase and composted solid digestate. In addition, the solid digestate was sampled at another plant. Results The plant, where the solid digestate originated from, and the soil type influenced nitrous oxide (N2O) emissions significantly over the 38-day experiment. Composting lowered N2O emissions after soil application, whereas bentonite addition did not have a significant effect. High peaks of N2O emissions were observed during the first days after application of acidified, liquid fraction of digestate. N2O emissions were strongly correlated to initial ammonium (NH4+) content. Conclusion Fertilization with dewatered digestate (both fractions) increased N2O emission, especially when applied to soils high in nutrients and organic matter.

Our previous studies demonstrated that it is possible to perform the classification of both simulated pressed and actual vocally fatigued voice productions versus vocally healthy productions through the pattern recognition of sEMG signals obtained from subjects’ anterior neck. In these studies, the commonly accepted Vocal Fatigue Index factor 1 (VFI-1) was used for the ground-truth labeling of normal versus vocally fatigued voice productions. Through recent experiments, other factors with potential effects on classification were also studied, such as sEMG signal normalization, and data imbalance—i.e., the large difference between the number of vocally healthy subjects and of those with vocal fatigue. Therefore, in this paper, we present a much improved classification method derived from an extensive study of the effects of such extrinsic factors on the classification of vocal fatigue. The study was performed on a large number of sEMG signals from 88 vocally healthy and fatigued subjects including student teachers and teachers and it led to important conclusions on how to optimize a machine learning approach for the early detection of vocal fatigue.

Bisphosphonates (BPs) are selective inhibitors of osteoclasts, used for the treatment of bone disorders. The objective of this study is to investigate the possible effects of BPs on the tongue's mucosa. Specimens of the tongue of 20 female 12-month old Wistar rats were taken. Ten were used as control group, while in the remaining alendronate (Fosamax, Merck) was administered per os from 13 weeks. Observation of the harvested samples was made by Transmission Electron Microscopy (TEM). In the experimental group, focal alterations were observed to various extent in all specimens. The basement membrane was intact. Furthermore, an increase at the intercellular space was observed, predominantly at the middle layer, and the desmosomes were disorganized. In the lamina propria focal edema was observed. Investigation on the effect of BPs on the tongue's mucosa through TEM hasn't been documented in the past. According to our results, BPs seem to cause mild mucosal lesions on the tongue.

Background and objectives: Bisphosphonates represent selective inhibitors of excess osteoblastic bone resorption that characterizes all osteopathies, targeting osteoclasts and their precursors. Their long-term administration in postmenopausal women suffering from osteoporosis has resulted in neural adverse effects. The current study focuses on the research of possible alterations in the femoral nerve, caused by bisphosphonates. We hypothesized that bisphosphonates, taken orally (per os), may produce degenerative changes to the femoral nerve, affecting lower-limb posture and walking neuronal commands. Materials and Methods: In order to support our hypothesis, femoral nerve specimens were extracted from ten female 12-month-old Wistar rats given 0.05 milligrams (mg) per kilogram (kg) of body weight (b.w.) per week alendronate per os for 13 weeks and from ten female 12-month-old Wistar rats given normal saline that were used as a control group. Specimens were studied using immunohistochemistry for selected antibodies NeuN (Neuronal Nuclear Protein), a protein located within mature, postmitotic neural nucleus, and cytosol and Sox10 (Sex-determining Region Y (SRY)—High-Motility Group (HMG)—box 10). The latter marker is fundamental for myelination of peripheral nerves. Obtained slides were examined under a light microscope. Results: Samples extracted from rats given alendronate were more Sox10 positive compared to samples of the control group, where the marker’s expression was not so intense. Both groups were equally NeuN positive. Our results are in agreement with previous studies conducted under a transmission electron microscope. Conclusions: The suggested pathophysiological mechanism linked to histological alterations described above is possibly related to toxic drug effects on Schwann and neuronal cells. Our hypothesis enhances the existing scientific evidence of degenerative changes present on femoral nerve following bisphosphonates administration, indicating a possible relationship between alendronate use and neuronal function.

Communication problems (eg, dysphonia, dysfluency and language and articulation disorders), swallowing disorders (dysphagia and globus), cough and upper airway symptoms, resulting from functional neurological disorder (FND), are commonly encountered by speech and language professionals. However, there are few descriptions in the literature of the most effective practical management approaches. This consensus document aims to provide recommendations for assessment and intervention that are relevant to both adults and young people. An international panel of speech and language professionals with expertise in FND were approached to take part. Participants responded individually by email to a set of key questions regarding best practice for assessment and interventions. Next, a video conference was held in which participants discussed and debated the answers to these key questions, aiming to achieve consensus on each issue. Drafts of the collated consensus recommendations were circulated until consensus was achieved. FND should be diagnosed on the basis of positive clinical features. Speech and language therapy for FND should address illness beliefs, self-directed attention and abnormal movement patterns through a process of education, symptomatic treatment and cognitive behavioural therapy within a supportive therapeutic environment. We provide specific examples of these strategies for different symptoms. Speech and language professionals have a key role in the management of people with communication and related symptoms of FND. It is intended that these expert recommendations serve as both a practical toolkit and a starting point for further research into evidence-based treatments.

Background Bisphosphonates (BPs) are forceful inhibitors of osteoclast-mediated bone resorption. Long-term BP use is associated with multiple rare but severe adverse effects. The objective of this study was to investigate the possible effects of BPs in the structure of femoral nerve. Specimens from the femoral nerve of ten female 12-month old Wistar rats were used as control group and ten female 12-month old Wistar rats to which Alendronate (Fosamax, Merck) was administered per os for 13 weeks, were used as research group. Samples were observed under a Transmission Electron Microscope. G ratio measurements and statistical analysis with SPSS program were also performed. Results The control group showed no major changes of the nerve’s histologic image, with the exception of some spots of thickness of the nerve myelin sheath. The research group showed major morphological changes which varied from partial disorganization or thickening of the myelin to severe myelin thickening and axon strangulation. A statistically significant difference of the G ratio between the two groups was observed. Conclusions The reported values (found in literature) for the morphologic measurements of the femoral nerve in Wistar rats are not complying with the ones we found in our study. There was a significant reduction of all three variables (the mean axon like diameter, the myelin thickness, G ratio) studied in the femoral nerve of the research group in contrast to control group. Our study demonstrates a possible correlation between alendronate administration and femoral nerve’s function, nevertheless due to the small specimen further research is needed.

Purpose: To examine the proposal that introversion predictably influences extralaryngeal and vocal behavior in vocally healthy individuals compared with individuals with extraversion and whether differences are of a nature that may support a risk hypothesis for primary muscle tension dysphonia. Method: Fifty-four vocally healthy female adults between the ages of 18 and 35 years were divided into 2 groups: introversion (n = 27) and extraversion (n = 27). All participants completed a psychological stress reactivity experiment. Before, during, and after the stressor (public speaking), participants were assessed on extralaryngeal muscle activity (surface electromyography: submental, infrahyoid; control site: tibialis anterior), perceived vocal effort, and vocal acoustics (fundamental frequency and intensity). Results: Participants in the introversion group exhibited significantly greater infrahyoid muscle activity throughout the protocol and during perceived stress than participants in the extraversion group. For both groups, perceived vocal effort significantly increased during stress, and acoustic measures significantly decreased. Infrahyoid muscle activity during the stress phase was significantly correlated with introversion and Voice Handicap Index scores but not with vocal effort scores. Conclusions: The data provided evidence of distinct differences in extralaryngeal behavior between introverts and extraverts. The findings are consistent with the trait theory of voice disorders (Roy & Bless, 2000).