Miracle-dz/UniFAR-FTUs huggingface dataset:
One-Pot Production of l-threo-3-Hydroxyaspartic Acid Using Asparaginase-Deficient Escherichia coli Expressing Asparagine Hydroxylase of Streptomyces coelicolor A3(2)

ABSTRACT We developed a novel process for efficient synthesis of l-threo-3-hydroxyaspartic acid (l-THA) using microbial hydroxylase and hydrolase. A well-characterized mutant of asparagine hydroxylase (AsnO-D241N) and its homologous enzyme (SCO2693-D246N) were adaptable to the direct hydroxylation of l-aspartic acid; however, the yields were strictly low. Therefore, the highly stable and efficient wild-type asparagine hydroxylases AsnO and SCO2693 were employed to synthesize l-THA. By using these recombinant enzymes, l-THA was obtained by l-asparagine hydroxylation by AsnO followed by amide hydrolysis by asparaginase via 3-hydroxyasparagine. Subsequently, the two-step reaction was adapted to one-pot bioconversion in a test tube. l-THA was obtained in a small amount with a molar yield of 0.076% by using intact Escherichia coli expressing the asnO gene, and thus, two asparaginase-deficient mutants of E. coli were investigated. A remarkably increased l-THA yield of 8.2% was obtained with the asparaginase I-deficient mutant. When the expression level of the asnO gene was enhanced by using the T7 promoter in E. coli instead of the lac promoter, the l-THA yield was significantly increased to 92%. By using a combination of the E. coli asparaginase I-deficient mutant and the T7 expression system, a whole-cell reaction in a jar fermentor was conducted, and consequently, l-THA was successfully obtained from l-asparagine with a maximum yield of 96% in less time than with test tube-scale production. These results indicate that asparagine hydroxylation followed by hydrolysis would be applicable to the efficient production of l-THA.
Effects of handedness (left vs right) and cannabis abuse on intermanual coordination and negative symptoms in schizophrenic patients of the paranoid type

Intermanual coordination as an index of interhemispheric transfer and negative symptoms were investigated in 50 left- and 42 right-handed schizophrenic inpatients of the paranoid type, also including drug abusers. The primary objective was to show that there were higher values in intermanual coordination and fewer manifestations of negative symptoms in the left-handed compared to the right-handed patients. This assumption was based on previous studies. Most importantly, right- and left-handed patients showed a different behaviour in intermanual coordination, when the duration of illness was taken into consideration. Thus, long-term left-handed paranoid patients performed better in intermanual coordination and showed fewer manifestations of negative symptoms than did long-term right-handed patients. These results were true for the large group of all patients, and among them for the subgroup of patients without drug abuse. Consequently, higher scores in intermanual coordination in left-handed patients may be related to a better interhemispheric crosstalk resulting in less pronounced negative symptoms. Secondary objectives assessed by explorative data analysis included the effects of cannabis abuse. While cannabis abuse may be more prevalent in left-handed patients, its effects may be more pronounced in right-handed patients, scoring higher in intermanual coordination and lower in manifestations of negative symptoms.
Effect of PCB Surface Modifications on the EMC-to-PCB Adhesion in Electronic Packages

The characteristics of interfacial adhesion between epoxy molding compound (EMC) and printed circuit board (PCB) were investigated. The surface conditions of solder resist (SR) layers, which were used as an outer skin of PCB, were varied within the range that would be encountered in the manufacturing process and reliability test conditions. First, the number of times of plasma treatment on the SR surfaces and the delay time prior to EMC molding on them were considered to examine the surface cleaning process and the aging effect, respectively, on adhesion. Second, moisture on the surfaces of PCB prior to EMC molding and moisture absorption and desorption at the interface were considered to investigate the environmental effect on adhesion. An unsymmetric double cantilever beam test method was devised by modifying the conventional symmetrical double cantilever beam. As a result, the phase angle of fracture could be controlled to achieve stable crack propagation along the desired interface, which enabled valid adhesion energy to be measured. The adhesion energy increased with plasma treatment by over 50%, from 55 to 86 J/m2. The improved adhesion was attributed to the increased the polar groups on the SR surface due to plasma treatment, which helped enhanced chemical bonding between the EMC resin and the SR resin. However, excessive plasma was counterproductive as it weakened the SR surface and caused cohesive crack propagation to occur within the SR layer. Adhesion remained nearly constant for delay time up to several hours between plasma treatment and EMC molding. However, small degradation of adhesion was observed when the delay time was extended to 12 h. Moisture on and in the SR material before EMC molding had a significant effect on adhesion. Absorbed moisture at the interface decreased the adhesion. However, when the moisture was baked out, adhesion was recovered almost to the original reference.
Comparing the Tuberculin Skin Test and T-SPOT.TB Blood Test in Children

BACKGROUND: Interferon-γ–release assays (IGRAs) have been developed for the diagnosis of tuberculosis infection, but few data are available for children. There currently is no reference standard for the diagnosis of tuberculosis infection. OBJECTIVE: To compare the performance of 1 IGRA, the T-SPOT.TB assay with the tuberculin skin test (TST) in children with different epidemiologic risk factors for tuberculosis. METHODS: We conducted a prospective study of 210 patients referred to 3 pediatric tuberculosis clinics, including those with no risk factors for tuberculosis (low risk, n = 27), risk factors but no identifiable source case (intermediate risk, n = 78), contact with a known source case (high risk, n = 74), and active disease (n = 31). Children were tested with TST and T-SPOT.TB. Concordance analyses were performed, and assay outcomes were modeled by multivariate logistic regression. RESULTS: For 13 children with culture-confirmed tuberculosis disease, sensitivity of TST and T-SPOT.TB was 77% and 92%, respectively, and concordance was 69%. For high-risk children, concordance was 94% for BCG-unimmunized children and 88% for BCG-immunized children. For intermediate-risk children, concordance was 74% for BCG-unimmunized children and 33% for BCG-immunized children. For low-risk children, concordance was 74% for BCG-unimmunized children and 20% for BCG-immunized children. Multivariate analysis revealed that contact with a source case was associated with T-SPOT.TB result, but age and BCG immunization were not. CONCLUSIONS: T-SPOT.TB is comparable to the TST in the diagnosis of tuberculosis disease and identification of high-risk children with tuberculosis infection and is more specific than the TST in children who have received the BCG vaccine.
Presumed non-accidental injury with retinal haemorrhages – findings from a tertiary referral centre in the United Kingdom

Aim: To describe the clinical and demographic features of patients with retinal haemorrhages from presumed non-accidental injury (NAI) at a tertiary referral centre in Leeds over a 2-year period. Methods: All patients with retinal haemorrhages from presumed NAI between 1 January 2007 and 31 December 2008 were retrospectively identified from the hospital RetCam® (Clarity Medical System, Pleasanton, CA, USA) database. Case-notes, fundus photographs and radiological studies were retrieved for all patients and examined. Results: Over the study period, 14 infants had retinal haemorrhages secondary to presumed NAI. All were male with a mean age of 18 ± 15 weeks (range 2–47) and came from areas with a mean Index of Multiple Deprivation (IMD 2007) rank of 34 ± 27% (range 0.97–68). Seizure/collapse was the reason for presentation in 71% (10/14). Retinal haemorrhages were bilateral in 64% (9/14) and unilateral in 36% (5/14). They were single-layered in 71% (10/14) and multi-layered in 29% (4/14). Subdural haemorrhages were found in 93% (13/14) and were symmetrical in 77% (10/13). Skeletal survey was positive in 28% (4/14). Conclusions: In the context of presumed NAI, there is a strong association between presence of retinal haemorrhages and the likelihood of underlying subdural haemorrhage. In this region, male infants under 12 months, from deprived areas, appear to constitute a vulnerable group.
Mechanism of extracellular calcium regulation of intestinal epithelial tight junction permeability: Role of cytoskeletal involvement

Recent studies suggest that an abnormal increase in intestinal tight junction (TJ) permeability may be an important etiologic factor in number of diseases including Crohn's disease, NSAID‐associated enteritis, and various infectious diarrheal syndromes. The intracellular processes involved in regulation of intestinal epithelial TJ permeability, however, remain poorly understood. In this study, we used cultured Caco‐2 intestinal epithelial cells to examine the intracellular processes involved in extracellular Ca++ modulation of intestinal epithelial monolayer TJ barrier. Incubation of the filter‐grown Caco‐2 intestinal monolayers in Ca++‐free solution (CFS), consisting of modified Krebs‐buffer solution containing 0 mM Ca++ and 1 mM EGTA, resulted in a rapid drop in Caco‐2 epithelial resistance and increase in epithelial permeability to paracellular markers mannitol and inulin, indicating an increase in TJ permeability. The increase in Caco‐2 TJ permeability was rapidly reversed by the re‐introduction of Ca++ (1.8 mM) into the incubation medium. The CFS‐induced increase in Caco‐2 TJ permeability was associated with separation of the cytoplasmic and transmembrane TJ proteins, ZO‐1 and occludin, and formation of large intercellular openings between the adjoining cells. The CFS‐induced modulation of TJ barrier was associated with activation of myosin light chain kinase (MLCK) activity and centripetal retraction of peri‐junctional actin and myosin filaments. The inhibition of CFS‐induced activation of Caco‐2 MLCK with MLCK inhibitor (ML‐7) prevented the CFS‐induced retraction of actin and myosin filaments and the subsequent alteration of TJ barrier function and structure. Our results suggested that the CFS‐induced alteration of TJ proteins and functional increase in TJ permeability was mediated by Caco‐2 MLCK activation and the resultant contraction of the peri‐junctionally located actin‐myosin filaments. Consistent with the role of MLCK in this process, selected inhibitors of Mg++‐myosin ATPase and metabolic energy, but not protein synthesis inhibitors, also prevented the CFS‐induced retraction of actin and myosin filaments and the subsequent increase in TJ permeability. In conclusion, our results indicate that extracellular Ca++ is crucial for the maintenance of intestinal epithelial TJ barrier function. The removal of extracellular Ca++ from the incubation medium causes activation of Caco‐2 MLCK, which in turn leads to an increase in intestinal monolayer TJ permeability. Microsc. Res. Tech. 51:156–168, 2000. © 2000 Wiley‐Liss, Inc.
Response of abducens internuclear neurons to axotomy in the adult cat

The highly specific projection of abducens internuclear neurons on the medial rectus motoneurons of the oculomotor nucleus constitutes an optimal model for investigating the effects of axotomy in the central nervous system. We have analyzed the morphological changes induced by this lesion on both the cell bodies and the transected axons of abducens internuclear neurons in the adult cat. Axotomy was performed by the transection of the medial longitudinal fascicle. Cell counts of Nissl‐stained material and calretinin‐immunostained abducens internuclear neurons revealed no cell death by 3 months postaxotomy. Ultrastructural examination of these cells at 6, 14, 24, and 90 days postaxotomy showed normal cytological features. However, the surface membrane of axotomized neurons appeared contacted by very few synaptic boutons compared to controls. This change was quantified by measuring the percentage of synaptic coverage of the cell bodies and the linear density of boutons. Both parameters decreased significantly after axotomy, with the lowest values at 90 days postlesion (∼70% reduction). We also explored axonal regrowth and the possibility of reinnervation of a new target by means of anterograde labeling with biocytin. At all time intervals analyzed, labeled axons were observed to be interrupted at the caudal limit of the lesion; in no case did they cross the scar tissue to reach the distal part of the tract. Nonetheless, a conspicuous axonal sprouting was present at the caudal aspect of the lesion site. Structures suggestive of axonal growth were found, such as large terminal clubs, from which short filopodium‐like branches frequently emerged. Similar findings were obtained after parvalbumin and calretinin immunostaining. At the electron microscopy level, biocytin‐labeled boutons originating from the sprouts appeared surrounded by either extracellular space, which was extremely dilated at the lesion site, or by glial processes. The great majority of labeled boutons examined were, thus, devoid of neuronal contact, indicating absence of reinnervation of a new target. Altogether, these data indicate that abducens internuclear neurons survive axotomy in the adult cat and show some form of axonal regrowth, even in the absence of target connection. J. Comp. Neurol. 427:370–390, 2000. © 2000 Wiley‐Liss, Inc.
Diagnostic boundaries, reasoning and depressive disorder, II. Application of a probabilistic model to the OPCS general population survey of psychiatric morbidity in Great Britain

Background. Reliable prevalence and risk estimation of psychiatric disorder is a cornerstone to achieving objectives in public health psychiatry. Research strategies have increasingly depended, therefore, upon the progressive evolution and refinement of diagnostic approaches designed to reflect better current knowledge concerning prognosis, course and outcome but essentially the need to improve agreement between users of the various schemes. Methods. This paper contrasts a conventional with a probabilistic approach to the diagnosis of depression based upon the OPCS United Kingdom National survey of psychiatric morbidity. The probabilistic approach, while designed to mimic current diagnostic practice in relation to the depressive disorders, naturally includes provision for the allocation of respondents on a scale of diagnostic uncertainty according to the severity of their presenting condition. Results. Findings are reported arising from the application of the probabilistic method to three areas of research interest in public health psychiatry, namely; an evaluation of additivity of event exposure and depressive morbidity, secondly use of the approach for investigating psychosocial models of depressive disorder and thirdly for assessing the agreement between depressive disorder when classified according to competing diagnostic schemes. Conclusions. The results show application of the probabilistic approach to provide a firm basis for achieving gains in both the stability and precision of risk profile estimation for depressive conditions.
Altered Expression and Splicing of ESRP1 in Malignant Melanoma Correlates with Epithelial–Mesenchymal Status and Tumor-Associated Immune Cytolytic Activity

Epithelial tumors are thought to recapitulate an epithelial–mesenchymal transition during the transformation process. By assessing ESRP1 transcripts in melanomas, associations were found between more mesenchymal states, immune cytolytic activity, and checkpoint expression that may have implications for immunotherapy. Melanoma is one of the major cancer types for which new immune-based cancer treatments have achieved promising results. However, anti–PD-1 and anti–CTLA-4 therapies are effective only in some patients. Hence, predictive molecular markers for the development of clinical strategies targeting immune checkpoints are needed. Using The Cancer Genome Atlas (TCGA) RNAseq data, we found that expression of ESRP1, encoding a master splicing regulator in the epithelial–mesenchymal transition (EMT), was inversely correlated with tumor-associated immune cytolytic activity. That association holds up across multiple TCGA tumor types, suggesting a link between tumor EMT status and infiltrating lymphocyte activity. In melanoma, ESRP1 mainly exists in a melanocyte-specific truncated form transcribed from exon 13. This was validated by analyzing CCLE cell line data, public CAGE data, and RT-PCR in primary cultured melanoma cell lines. Based on ESRP1 expression, we divided TCGA melanoma cases into ESRP1-low, -truncated, and –full-length groups. ESRP1-truncated tumors comprise approximately two thirds of melanoma samples and reside in an apparent transitional state between epithelial and mesenchymal phenotypes. ESRP1 full-length tumors express epithelial markers and constitute about 5% of melanoma samples. In contrast, ESRP1-low tumors express mesenchymal markers and are high in immune cytolytic activity as well as PD-L2 and CTLA-4 expression. Those tumors are associated with better patient survival. Results from our study suggest a path toward the use of ESRP1 and other EMT markers as informative biomarkers for immunotherapy. Cancer Immunol Res; 4(6); 552–61. ©2016 AACR.
TLS and energy consumption on a mobile device: A measurement study

We report results from a measurement study on the role of the most popular end-to-end security protocol Transport Layer Security (TLS) in the energy consumption of a mobile device. We measured energy consumed by TLS transactions between a Nokia N95 and several popular Web services over WLAN and 3G network interfaces. Our detailed analysis corroborates some earlier results but also reveals, contrary to earlier studies, that the transmission and I/O energy, both in the TLS handshake and the record protocol, far exceed the required computational energy by the actual cryptographic algorithms and that with transactions larger than 500KB, the energy required to transmit the actual data clearly outranks the TLS energy overhead. In addition, we note that the energy consumption varies remarkably between measured services.
The short inflammatory bowel disease questionnaire is reliable and responsive to clinically important change in ulcerative colitis

OBJECTIVE:The Short Inflammatory Bowel Disease Questionnaire (SIBDQ) is a health-related quality of life (HRQoL) tool measuring physical, social, and emotional status (score 10–70, poor to good HRQoL). The SIBDQ has been predominantly used in trials for Crohn's disease, and further validation of the SIBDQ is desirable in ulcerative colitis (UC) patients. The primary objective was to further validate the SIBDQ by examining discriminant ability against measures of disease activity. The secondary objectives were to examine reliability and responsiveness to change.METHODS:UC patients attending hospital completed the SIBDQ and two activity indices. Patients’ disease status (remission, mild, moderate, or severe relapse) was determined subjectively by the patients and their physician.RESULTS:Scores were obtained for 69 events in 61 patients, mean age 47.8 yr (range 16–79). All classes of disease extent were represented. The mean SIBDQ score was 48.4 (13–70). The difference between mean score in patients in remission and relapse was −20.1 (95% CI =−25.1 to −15.1). The difference for remission and mild relapse was −14.6 (95% CI =−8.9 to −20.2). The correlation between SIBDQ and the activity indices were good, r = −0.83 and r = −0.61. Eight patients presented twice. Those with unchanged disease status showed no significant difference in the mean SIBDQ score. Patients whose disease status had deteriorated from remission to mild relapse, or from mild to moderate relapse demonstrated a mean reduction of 11.8 points (95% CI = 20.1–3.4). CONCLUSIONS:This study contributes to the validation of the SIBDQ as a HRQoL tool in UC. It is reproducible and responsive to changes in disease activity.
Impact of Pediatric Epilepsy on the Family: A New Scale for Clinical and Research Use

Summary: Purpose: There is no brief, global instrument available that specifically measures the psychosocial impact of pediatric epilepsy on the family. An 11‐item scale was created for parents' use to evaluate the influence of epilepsy on the major aspects of their family and child's life. This will be a helpful tool both in the clinical setting and as an outcome measure for research studies.
Experiences of using an Environmental Control System (ECS) for persons with high cervical spinal cord injury: the interplay between hassle and engagement

Abstract Purpose: Environmental Control Systems (ECS) have many benefits; however studies of personal experience of ECS use are scarce. This qualitative study explored the insiders’ experience of using an ECS. Method: An ECS starter-pack was compiled and trialled for an eight-week period with six persons with high spinal cord injuries (SCI) living in Ireland. Semi-structured interviews were subsequently completed with each person and analysed using Interpretative Phenomenological Analysis (IPA). Findings: Two major themes emerged: “Taking back a little of what has been lost”, and “Getting used to ECS” which is the focus of the current paper. This theme captured a dynamic interplay between the experience of “hassle” and “engagement” for new users of ECS. “Hassle” resulted from technological frustrations and the challenge of breaking familiar habits, while “Engagement” resulted from feeling good, having fun and being surprised. Conclusions: The complex interweaving of hassle and engagement experienced by new ECS users reflects the clinical experience of rehabilitation providers. The importance of overcoming initial hassle needs to be understood by clinicians and users in order to maximise the potential benefit of ECS. Non-use must be considered one reasonable outcome if based on realistic ECS trials. Implications for Rehabilitation People with high cervical SCI find ECS use to be engaging, enjoyable, and fun. Successful engagement with ECS involves overcoming “hassle”. For some, ECS may be too much hassle resulting in non-use. However, it is important that non-use is not necessarily considered a failure, provided it is based on real experience and well-informed choice. Realistic trials need to be made available early in rehabilitation in order to facilitate the establishment of ECS use as a habit.