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OaPAC is a recently discovered blue-light-using flavin adenosine dinucleotide (BLUF) photoactivated adenylate cyclase from the cyanobacterium Oscillatoria acuminata that uses adenosine triphosphate and translates the light signal into the production of cyclic adenosine monophosphate. Here, we report crystal structures of the enzyme in the absence of its natural substrate determined from room-temperature serial crystallography data collected at both an X-ray free-electron laser and a synchrotron, and we compare these structures with cryo-macromolecular crystallography structures obtained at a synchrotron by us and others. These results reveal slight differences in the structure of the enzyme due to data collection at different temperatures and X-ray sources. We further investigate the effect of the Y6W mutation in the BLUF domain, a mutation which results in a rearrangement of the hydrogen-bond network around the flavin and a notable rotation of the side chain of the critical Gln48 residue. These studies pave the way for picosecond–millisecond time-resolved serial crystallography experiments at X-ray free-electron lasers and synchrotrons in order to determine the early structural intermediates and correlate them with the well studied picosecond–millisecond spectroscopic intermediates.

Students learn not only directly from their teachers and books, but also by using their computers, tablets, and phones. Monitoring these learning environments creates new opportunities for teachers to track students' progress. In particular, this article is based on gathering real-time events as students interact with learning tools and materials in electronic devices, both in and out of class. Our study shows that the analysis of these events can provide teachers with week-by-week predictions of their students' final grades and help them to identify, at an early stage, those students at risk of failing. A blended environment university course in which students are expected to work autonomously out of class, but also attend face-to-face lessons was used as the case study. Results show that predictions are reasonably accurate even during the first weeks of the course.

ABSTRACTIntroduction Globally, diarrheal disease continues to be the second leading cause of death among under-five children, especially in developing countries. This study aimed to determine the prevalence of diarrhea and associated risk factors among under-five attending Robe Didea General Hospital (RDGH) in Arsi Robe Town, Oromia Regional State, Ethiopia, from October 1 to December 30 2020.Methods A hospital-based cross-sectional study was conducted among under-five children who attended the pediatrics department of RDGH. Systematic random sampling was employed to select the study subjects. An interviewer-administered structured questionnaire was used for data collection and the analysis used SPSS version 21. Bivariate and multivariable logistic regression analyses were employed to determine the association between risky factors and diarrheal prevalence, and a p value

ABN (Abierto Basado en Números--Open Calculation Based on Numbers) is a method for teaching basic arithmetic operations in primary education that has become popular in recent years and that is based on the decomposition of numbers through manipulative materials that encourage mental calculation. Currently there is limited number of tools that can be used to support the ABN method and so this article presents the design and development of two tools that facilitate learning with this method, a physical device, ABENEARIO-P, and a virtual device (web application), ABENEARIO-V, that complements the physical device. In addition, a study of the use of these tools was carried out with 80 learners (ages 7 and 9) and 9 teachers with a focus on ABENEARIO-V. The results of this study showed a positive evaluation of the tool by both learners and teachers, an adequate time to complete the mathematical operations assigned to learners and an improvement in performance as the tool was used. As a conclusion, it is important to provide adequate tools that can support teachers and learners in the practice with the ABN method as in the case of ABENEARIO-P and ABENEARIO-V. Limitations refer mainly to the context of the study, which was conducted at a time of severe social distance restrictions during the COVID-19 pandemic on touching physical devices or being able to gather a larger number of learners in the classroom.

The fluorescent glutamate indicator iGluSnFR enables imaging of neurotransmission with genetic and molecular specificity. However, existing iGluSnFR variants exhibit low in vivo signal-to-noise ratios, saturating activation kinetics and exclusion from postsynaptic densities. Using a multiassay screen in bacteria, soluble protein and cultured neurons, we generated variants with improved signal-to-noise ratios and kinetics. We developed surface display constructs that improve iGluSnFR's nanoscopic localization to postsynapses. The resulting indicator iGluSnFR3 exhibits rapid nonsaturating activation kinetics and reports synaptic glutamate release with decreased saturation and increased specificity versus extrasynaptic signals in cultured neurons. Simultaneous imaging and electrophysiology at individual boutons in mouse visual cortex showed that iGluSnFR3 transients report single action potentials with high specificity. In vibrissal sensory cortex layer 4, we used iGluSnFR3 to characterize distinct patterns of touch-evoked feedforward input from thalamocortical boutons and both feedforward and recurrent input onto L4 cortical neuron dendritic spines.

Student outcomes depend on the implementation of intervention practice elements, which are the specific behaviors that front-line implementers deliver as part of their direct interactions with students. Tier 1 PBIS is a well established practice in schools but existing guidance on practice elements is often conflated with guidance on organizational systems that support implementation. The goal of this study was to distill the specific practice elements of Tier 1 PBIS from the established core features using an expert consensus process. A panel of nine Tier 1 PBIS research and practice experts participated in three iterative rounds of information gathering and feedback within an electronic Delphi study that structured the consensus-building process. The expert panel reached consensus by distilling Tier 1 PBIS into 16 distinct practice elements organized into five thematic domains. The thematic domains from the present study align with existing literature on core Tier 1 PBIS features but the practices in each domain clearly define the specific, expected actions of frontline implementers. Implications are discussed in terms of how the identified practice elements may be used to inform intervention fidelity monitoring and targeted implementation supports for front-line implementers.

Check-in/Check-out (CICO) is a widely implemented evidence-based program for supporting students with at-risk levels of social and emotional behavior concerns. It is comprised of several core features described in the previous literature, including "practice elements," which are the specific actions that are delivered directly to students, and "implementation components," which are actions that support the implementation by adults. Practice elements and implementation components are both important to implementation but have been combined and conflated in descriptions of CICO implementation. Well-defined and differentiated practice elements could provide improved clarity in communicating implementation expectations to frontline implementers as well as support future research into essential active ingredients and measures of front-line intervention fidelity. The purpose of the present study was to distill, differentiate, and operationally define the student-facing practice elements of CICO. A panel of research experts and practice experts participated in a three-round modified e-Delphi process that led to the identification and operational definition of 19 discreet practice elements organized into five domains. Results are discussed in terms in implications for future development of measures of commitment and intervention fidelity, future research into active ingredients of CICO, and in terms of how well-defined practice elements can improve communication of implementation expectations for front-line implementers of CICO such as teachers.

Abstract Background Disparities in child healthcare service utilization are unacceptably high in Ethiopia. Nevertheless, little is known about underlying barriers to accessing child health services, especially among low socioeconomic subgroups and in remote areas. This study aims to identify barriers to equity in the use of child healthcare services in Ethiopia. Methods Data were obtained from 20 key- informant interviews (KII) and 6 focus group discussions (FGD) with mothers and care givers. This study was conducted in Oromia Region, Arsi Zone, Zuway Dugda District from June 1–30, 2023. The study participants for this research were selected purposively. The information was collected based on the principle of saturation after sixteen consecutives interview were conducted. Both KII and FGD were audio-recorded and complementary notes were taken to record observations about the participants’ comments and their interactions. Each interview and FGD data were transcribed word-for-word in the local Afaan Oromo and Amaharic languages and then translated to English language. Finally, the data were analyzed thematically using NVivo 14 software and narrated in the linked pattern of child health service utilization. Results This study identified six major themes which emerged as barriers to healthcare utilization equity for caregivers and their -under-five children. Barriers related to equity in low level of awareness regarding need, low socioeconomic status, geographical inaccessibility, barriers related to deficient healthcare system, community perception and cultural restrictions, and barriers of equity related to political instability and conflict. The most commonly recognized barriers of equity at the community level were political instability, conflict, and a tremendous distance to a health facility. Transportation challenges, poor functional services, closure of the health facility in working hours, and lack of proper planning to address the marginalized populations were identified barriers of equity at organizational or policy level. Conclusion This study showed that inequity in child healthcare utilization is an important challenge confronting Ethiopia. To achieve equity, policy makers and planners need to change health policy and structure to be pro-poor. It is also necessary to improve the healthcare system to increase service utilization and access for impoverished women, individuals with lower levels of education, and residents of isolated rural areas. Furthermore, context specific information pertaining to cultural barriers and political ecology are required.

Abstract The main protease (Mpro) of SARS-CoV-2 is critical for viral function and a key drug target. Mpro is only active when reduced; turnover ceases upon oxidation but is restored by re-reduction. This suggests the system has evolved to survive periods in an oxidative environment, but the mechanism of this protection has not been confirmed. Here, we report a crystal structure of oxidized Mpro showing a disulfide bond between the active site cysteine, C145, and a distal cysteine, C117. Previous work proposed this disulfide provides the mechanism of protection from irreversible oxidation. Mpro forms an obligate homodimer, and the C117-C145 structure shows disruption of interactions bridging the dimer interface, implying a correlation between oxidation and dimerization. We confirm dimer stability is weakened in solution upon oxidation. Finally, we observe the protein’s crystallization behavior is linked to its redox state. Oxidized Mpro spontaneously forms a distinct, more loosely packed lattice. Seeding with crystals of this lattice yields a structure with an oxidation pattern incorporating one cysteine-lysine-cysteine (SONOS) and two lysine-cysteine (NOS) bridges. These structures further our understanding of the oxidative regulation of Mpro and the crystallization conditions necessary to study this structurally.

Heroda Gebru,1 Negussie Deyessa,2 Girmay Medhin,3 Helmut Kloos4 1Ethiopian Institute of Water Resources, Addis Ababa University, Addis Ababa, Ethiopia; 2School of Public Health, College of Health Sciences, Addis Ababa University, Addis Ababa, Ethiopia; 3Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia; 4Department of Epidemiology and Biostatistics, University of California, San Francisco, CA, USACorrespondence: Heroda Gebru, Email herodagebru13@gail.comBackground: The recurrence of intestinal parasitic infections (IPIs) can lead to different problems that can be transferred from generation to generation. Sanitation and hygienic practices have vital role in the parasitic reinfection. In poor hygienic and sanitation condition children may live in a continuous cycle of infection and reinfection.Objective: To assess childhood IP reinfection and its association with sanitation and hygienic practice in eastern Ethiopia.Methods: A population-based case-control design was used in this study. Data were collected from 75 reinfected cases and 147 unmatched controls. Fecal specimens were observed for parasites using direct smear and formol ether techniques. Epi-Info and SPSS (the statistical package for social science) were used for data entry and analysis, respectively. Logistic regression analysis was conducted to identify significant associations (P< 0.05) between variables.Results: The overall IP reinfection rate within 24 weeks after treatment was 33.8% (75/222), with a 95% CI=27.7%– 40.5%. The frequency of intestinal protozoa was 18%, and for helminths was 15.8%. Children who swam in a polluted water had 3.7 times greater odds of IP reinfection than children who did not swim (P =0 .01, 95% CI: 1.4– 10.0). Children who regularly bathed in streams and children who bathed both at home and in streams were found to have 12.6 times and 5.8 times higher odds of IP reinfection than children who bathed regularly at home (P=0.002, 95% CI:2.5– 64.8) and (P = 0.042, 95% CI:1.1– 31.3), respectively. Children in households that owned domestic animals had 4.5 times higher odds of IP reinfection than the reference group (P = 0.013, 95% CI: 1.3– 12.5).Conclusion: IP reinfection rates were significantly associated with habits of swimming in a polluted water, places of bathing, and ownership of domestic animals. Therefore, efforts should be made considering such factors to minimize IP reinfection in the area.Keywords: Ethiopia, hygiene, reinfection, sanitation

Educational content has become a key element for improving the quality and effectiveness of teaching. Many studies have been conducted on user and knowledge modeling using machine-learning algorithms in smart-learning environments. However, few studies have focused on content modeling to estimate content indicators based on student interaction. This study presents a systematic literature review of content modeling using machine learning algorithms in smart learning environments. Two databases were used: Scopus and Web of Science (WoS), with studies conducted until August 2023. In addition, a manual search was performed at conferences and in relevant journals in the area. The results showed that assessment was the most used content in the papers, with difficulty and discrimination as the most common indicators. Item Response Theory (IRT) is the most commonly used technique; however, some studies have used different traditional learning algorithms such as Random Forest, Neural Networks, and Regression. Other indicators, such as time, grade, and number of attempts, were also estimated. Owing to the few studies on content modeling using machine learning algorithms based on interactions, this study presents new lines of research based on the results obtained in the literature review.

Introduction: It is known that cognitive deficits are a core feature of schizophrenia and that in the general population, prior beliefs significantly influence learning and reasoning processes. However, the interaction of prior beliefs with cognitive deficits and their impact on performance in schizophrenia patients is still poorly understood. This study investigates the role of beliefs and cognitive variables (CVs) like working memory, associative learning, and processing speed on learning processes in individuals with schizophrenia. We hypothesize that beliefs will influence the ability to learn correct predictions and that first-episode schizophrenia patients (FEP) will show impaired learning due to cognitive deficits. Methods: We used a predictive-learning task to examine how FEP (n = 23) and matched controls (n = 23) adjusted their decisional criteria concerning physical properties during the learning process when predicting the sinking behavior of two transparent containers filled with aluminum discs when placed in water. Results: On accuracy, initial differences by group, trial type, and interaction effects of these variables disappeared when CVs were controlled. The differences by conditions, associated with differential beliefs about why the objects sink slower or faster, were seen in patients and controls, despite controlling the CVs' effect. Conclusions: Differences between groups were mainly explained by CVs, proving that they play an important role than what is assumed in this type of task. However, beliefs about physical events were not affected by CVs, and beliefs affect in the same way the decisional criteria of the control or FEP patients' groups.

Potential benefits of learning analytics (LA) for improving students' performance, predicting students' success, and enhancing teaching and learning practice have increasingly been recognized in higher education. However, the adoption of LA in higher education institutions (HEIs) to date remains sporadic and predominantly small in scale due to several socio-technical challenges. To better understand why HEIs struggle to scale LA adoption, it is needed to untangle adoption challenges and their related factors. This paper presents the findings of a study that sought to investigate the associations of adoption factors with challenges HEIs face in the adoption of LA and how these associations are compared among HEIs at different scopes of adoption. The study was based on a series of semi-structured interviews with senior managers in HEIs. The interview data were thematically analysed to identify the main challenges in LA adoption. The connections between challenges and other factors related to LA adoption were analysed using epistemic network analysis (ENA). From senior managers' viewpoints, ethical issues of informed consent and resistance culture had the strongest links with challenges of learning analytic adoption in HEI; this was especially true for those institutions that had not adopted LA or who were in the initial phase of adoption (i.e., preparing for or partially implementing LA). By contrast, among HEIs that had fully adopted LA, the main challenges were found to be associated with centralized leadership, gaps in the analytic capabilities, external stakeholders, and evaluations of technology. Based on the results, we discuss implications for LA strategy that can be useful for institutions at various stages of LA adoption, from early stages of interest to the full adoption phase.

Reversibly photoswitchable fluorescent proteins are essential markers for advanced biological imaging, and optimization of their photophysical properties underlies improved performance and novel applications. Here we establish a link between photoswitching contrast, one of the key parameters that dictate the achievable resolution in nanoscopy applications, and chromophore conformation in the non-fluorescent state of rsEGFP2, a widely employed label in REversible Saturable OpticaL Fluorescence Transitions (RESOLFT) microscopy. Upon illumination, the cis chromophore of rsEGFP2 isomerizes to two distinct off-state conformations, trans1 and trans2, located on either side of the V151 side chain. Reducing or enlarging the side chain at this position (V151A and V151L variants) leads to single off-state conformations that exhibit higher and lower switching contrast, respectively, compared to the rsEGFP2 parent. The combination of structural information obtained by serial femtosecond crystallography with high-level quantum chemical calculations and with spectroscopic and photophysical data determined in vitro suggests that the changes in switching contrast arise from blue- and red-shifts of the absorption bands associated to trans1 and trans2, respectively. Thus, due to elimination of trans2, the V151A variants of rsEGFP2 and its superfolding variant rsFolder2 display a more than two-fold higher switching contrast than their respective parent proteins, both in vitro and in E. coli cells. The application of the rsFolder2-V151A variant is demonstrated in RESOLFT nanoscopy. Our study rationalizes the connection between structural and photophysical chromophore properties and suggests a means to rationally improve fluorescent proteins for nanoscopy applications.

Background: Cytopathological evaluation of thyroid fine-needle aspiration biopsy (FNAB) specimens can fail to raise preoperative suspicion of medullary thyroid carcinoma (MTC). The Afirma RNA-sequencing MTC classifier identifies MTC among FNA samples that are cytologically indeterminate, suspicious, or malignant (Bethesda categories III-VI). In this study we report the development and clinical performance of this MTC classifier. Methods: Algorithm training was performed with a set of 483 FNAB specimens (21 MTC and 462 non-MTC). A support vector machine classifier was developed using 108 differentially expressed genes, which includes the 5 genes in the prior Afirma microarray-based MTC cassette. Results: The final MTC classifier was blindly tested on 211 preoperative FNAB specimens with subsequent surgical pathology, including 21 MTC and 190 non-MTC specimens from benign and malignant thyroid nodules independent from those used in training. The classifier had 100% sensitivity (21/21 MTC FNAB specimens correctly called positive; 95% confidence interval [CI] = 83.9-100%) and 100% specificity (190/190 non-MTC FNAs correctly called negative; CI = 98.1-100%). All positive samples had pathological confirmation of MTC, while all negative samples were negative for MTC on surgical pathology. Conclusions: The RNA-sequencing MTC classifier accurately identified MTC from preoperative thyroid nodule FNAB specimens in an independent validation cohort. This identification may facilitate an MTC-specific preoperative evaluation and resulting treatment.

Objective: In patients undergoing elective thoracic surgery, frailty is associated with worse outcomes. However, the magnitude by which frailty influences outcomes of urgent thoracic surgery (UTS) is unknown. Methods: We identified patients admitted with a UTS condition from January to September 2017 in the National Readmissions Database. UTS conditions were classified as esophageal perforation, hemo/pneumothorax, rib fracture, and obstructed hiatal hernia. Outcome of interest was mortality within 90 days of index admission. Frailty score was calculated using a deficit accumulation method. Cox proportional hazard modeling was used to calculate a hazard ratio for each combination of UTS disease type and frailty score, adjusted for sex, insurance payor, hospital size, and hospital and patient location, and was compared with the effect of frailty on elective lung lobectomy. Results: We identified 107,487 patients with a UTS condition. Among UTS conditions overall, increasing frailty elements were associated with increased mortality (hazard ratio, 2270; 95% CI, 1463-3523; P

This research examines the implications of the nyikok tradition in Lampung Pepadun customary society on the fulfillment of children's rights and its relevance to the development of Islamic family law in Indonesia. This type of research is field research, which is conducted in the Lampung Pepadun indigenous community in Lampung Province. This type of research is field research, which is conducted in the Lampung Pepadun Customary Society in Lampung Province. Data sources in this research are primary and secondary data sources. Data analysis in this research uses qualitative methods. Analyzed inductive thinking approach. The results of the study that Nyikok in Lampung Pepadun cultural customs has a good purpose. Based on the Receptio A Contrario analysis, nyikok has compatibility with Islamic law, so that its practice can be applied by the indigenous people of Lampung Pepadun, because it has positive implications in fulfilling children's rights, on the other hand, it is also to protect the inherited property of the deceased father, to continue the breed so that the balance is not broken and to maintain the kinship system that has been established. The nyikok tradition has relevance to Islamic family law in Indonesia, which is relevant to KHI Article 14, Article 2 (1) of Law No. 1 of 1974 concerning Marriage and then the existence of nyikok in the context of child protection and care is relevant to Article 45 of Law No. 1 of 1974 concerning Marriage, that parents have an obligation to maintain and educate children. Keyword: Nyikok; Lampung Pepadun; Children's Rights; Indonesian Family Law.

ABSTRACTIntroduction Tuberculosis remains a major global health problem and ranks alongside the human immunodeficiency virus (HIV) as a leading cause of mortality worldwide. This study investigated the treatment outcome of tuberculosis and predictors of unsuccessful treatment outcome of tuberculosis (TB) patients enrolled in Arsi-Robe Hospital, central Ethiopia between January 2013 and December 2017.Methods An institution-based retrospective study was conducted on patients who had all forms of TB such as smear positive tuberculosis (PTB+), smear negative tuberculosis (PTB-), and extrapulmonary tuberculosis (EPTB) in the DOTS clinic. A multivariable logistic regression model was employed to identify predictors of unsuccessful treatment outcome and a P

Effective pain management in laboratory animals is crucial for both animal welfare and the reliability of scientific research. We retrospectively examined the effects of carprofen as post-operative analgesia in Sprague Dawley rats following stereotactic surgery. Our data indicate that administering carprofen twice daily (5 mg/kg), as currently recommended by Die Gesellschaft für Versuchstierkunde/Society for Laboratory Animal Science (GV-SOLAS), led to adverse effects such as reduced food and water intake, disrupted fecal excretion, and abdominal bloating consistent with peritonitis. Continued administration exacerbated these symptoms, with post-mortem findings of intestinal obstructions and ulcers. However, when the frequency was reduced to once daily, such adverse symptoms were not observed. These results are based on incidental data collected from various neuroscientific experiments, resulting in small and uneven sample groups across various experimental cohorts. The inherent imbalances in these groups present challenges for statistical interpretation. While the findings suggest that less frequent carprofen use may reduce adverse effects, the surgical interventions and concurrent use of other drugs in these experiments likely exacerbated these outcomes. Further investigation into the interactions between carprofen, surgical stress, and other perioperative factors is needed to refine analgesia protocols in laboratory animals. Despite these limitations, these observations contribute to understanding analgesia protocols and may assist in improving animal welfare practices.