Miracle-dz/UniFAR-FTUs huggingface dataset:
Comparison of family 12 glycoside hydrolases and recruited substitutions important for thermal stability

As part of a program to discover improved glycoside hydrolase family 12 (GH 12) endoglucanases, we have studied the biochemical diversity of several GH 12 homologs. The H. schweinitzii Cel12A enzyme differs from the T. reesei Cel12A enzyme by only 14 amino acids (93% sequence identity), but is much less thermally stable. The bacterial Cel12A enzyme from S. sp. 11AG8 shares only 28% sequence identity to the T. reesei enzyme, and is much more thermally stable. Each of the 14 sequence differences from H. schweinitzii Cel12A were introduced in T. reesei Cel12A to determine the effect of these amino acid substitutions on enzyme stability. Several of the T. reesei Cel12A variants were found to have increased stability, and the differences in apparent midpoint of thermal denaturation (Tm) ranged from a 2.5°C increase to a 4.0°C decrease. The least stable recruitment from H. schweinitzii Cel12A was A35S. Consequently, the A35V substitution was recruited from the more stable S. sp. 11AG8 Cel12A and this T. reesei Cel12A variant was found to have a Tm 7.7°C higher than wild type. Thus, the buried residue at position 35 was shown to be of critical importance for thermal stability in this structural family. There was a ninefold range in the specific activities of the Cel12 homologs on o‐NPC. The most and least stable T. reesei Cel12A variants, A35V and A35S, respectively, were fully active. Because of their thermal tolerance, S. sp. 11AG8 Cel12A and T. reesei Cel12A variant A35V showed a continual increase in activity over the temperature range of 25°C to 60°C, whereas the less stable enzymes T. reesei Cel12A wild type and the destabilized A35S variant, and H. schweinitzii Cel12A showed a decrease in activity at the highest temperatures. The crystal structures of the H. schweinitzii, S. sp. 11AG8, and T. reesei A35V Cel12A enzymes have been determined and compared with the wild‐type T. reesei Cel12A enzyme. All of the structures have similar Cα traces, but provide detailed insight into the nature of the stability differences. These results are an example of the power of homolog recruitment as a method for identifying residues important for stability.
Genetic diversity and dynamics of Plasmodium falciparum and P. vivax populations in multiply infected children with asymptomatic malaria infections in Papua New Guinea

We describe the dynamics of co-infections of Plasmodium falciparum and P. vivax in 28 asymptomatic children by genotyping these species using the polymorphic loci Msp2 and Msp3α, respectively. The total number of Plasmodium spp. infections detected using 3 day sampling over 61 days varied between 1 and 14 (mean 6·6). The dynamics of P. falciparum and P. vivax genotypes varied greatly both within and amongst children. Periodicity in the detection of P. falciparum infections is consistent with the synchronous replication of individual genotypes. Replication synchrony of multiple co- infecting genotypes was not detected. In 4-year-old children P. falciparum genotype complexity was reduced and episodes lasted significantly longer (median duration > 60 days) when compared to children aged 5–14 years (median duration 9 days). P. vivax genotype complexity was not correlated with age but the episode duration was also longer for this species in 4-year-olds than in older children but was not as long as P. falciparum episodes. Recurrence of P. falciparum and P. vivax genotypes over weeks was observed. We interpret these major fluctuations in the density of genotypes over time as the result of the mechanism of antigenic variation thought to be present in these Plasmodium species.
PSYCHOLOGY, PSYCHIATRY & BRAIN NEUROSCIENCE SECTION Original Research Articles Psychological Flexibility May Reduce Insomnia in Persons with Chronic Pain: A Preliminary Retrospective Study

Objective. Sleep disturbance is a common com- plaint in people with chronic pain, and is associated with a range of adverse outcomes including reports of greater pain and disability. Research into insom- nia with chronic pain sufferers has not yet examined the role of psychological flexibility, a process from acceptance and commitment therapy. We examined this role. Design. Participants in this study were 159 adult patients attending an assessment at a specialist pain center. They were mostly women, 63%, and the mean age was 43.8 years. Results. In preliminary analyses 79% of partici- pants met the criteria for significant insomnia based on their self-reported symptoms. As predicted, sig- nificant positive correlations were found between components of psychological flexibility, particularly acceptance of pain and values-based action, and all measures of sleep quality. In regression analyses, the components of psychological flexibility consid- ered together accounted for between 11% and 19% of variance across a range of measures of sleep quality. Conclusion. These results suggest the need to further develop treatment services for people with chronic pain and insomnia.
Real-time ultrasonic observation of combined spinal-epidural anaesthesia

Background and objective: The quality of combined spinal-epidural anaesthesia mainly depends on accurate identification of the epidural space. The real-time ultrasound control of the procedure for puncture was therefore evaluated. Methods: Thirty parturients scheduled for Caesarean section were randomized to three equal groups. Ten control patients received conventional combined spinal-epidural anaesthesia. Ten of the remaining patients received ultrasonic scans by an offline scan technique, and 10 received online imaging of the lumbar region during epidural puncture. The epidural space was identified and needle advancement was surveyed through the interspinal and flaval ligaments. The number of attempts to advance the needle to achieve a successful puncture was measured and compared, as well as the number of vertebral interspaces punctured before successful entry into the epidural space. Results: There was no difference between patient characteristics in the three groups. The visualization of the epidural structures and of the needle manipulations was very effective. In the ultrasound group, the reduction in the number of attempts at puncture was significant (P < 0.036). The number of interspaces necessary for puncture was reduced (P < 0.036) in the ultrasound online group compared with controls. The number of spinal needle manipulations was significantly reduced (P < 0.036). Conclusions: Real-time ultrasonic scanning of the lumbar spine is an easy procedure. It provides an accurate reading of the location of the needle tip and facilitates the performance of combined spinal-epidural anaesthesia.
Growth Hormone Induces Hepatic Production of Fibroblast Growth Factor 21 through a Mechanism Dependent on Lipolysis in Adipocytes*

Background: Both growth hormone (GH) and FGF21 are fasting-inducible hormones involved in modulation of lipolysis. Results: GH injection caused a rapid elevation of circulating FGF21 in mice, and such an effect was blocked by the lipolysis inhibitor niacin. FGF21 knock-out mice exhibited greater potency in GH-induced lipolysis. Conclusion: GH and FGF21 form a negative feedback loop to fine-tune lipolysis in adipocytes. Significance: This study provides a novel insight on hormonal control of lipolysis. Fibroblast growth factor (FGF) 21 and growth hormone (GH) are metabolic hormones that play important roles in regulating glucose and lipid metabolism. Both hormones are induced in response to fasting and exert their actions on adipocytes to regulate lipolysis. However, the molecular interaction between these two hormones remains unclear. Here we demonstrate the existence of a feedback loop between GH and FGF21 on the regulation of lipolysis in adipocytes. A single bolus injection of GH into C57 mice acutely increases both mRNA and protein expression of FGF21 in the liver, thereby leading to a marked elevation of serum FGF21 concentrations. Such a stimulatory effect of GH on hepatic FGF21 production is abrogated by pretreatment of mice with the lipolysis inhibitor niacin. Direct incubation of either liver explants or human HepG2 hepatocytes with GH has no effect on FGF21 expression. On the other hand, FGF21 production in HepG2 cells is significantly induced by incubation with the conditioned medium harvested from GH-treated adipose tissue explants, which contains high concentrations of free fatty acids (FFA). Further analysis shows that FFA released by GH-induced lipolysis stimulates hepatic FGF21 expression by activation of the transcription factor PPARα. In FGF21-null mice, both the magnitude and duration of GH-induced lipolysis are significantly higher than those in their wild type littermates. Taken together, these findings suggest that GH-induced hepatic FGF21 production is mediated by FFA released from adipose tissues, and elevated FGF21 in turn acts as a negative feedback signal to terminate GH-stimulated lipolysis in adipocytes.
Impaired Haemostasis by Intravenous Administration of a Gelatin-based Plasma Expander in Human Subjects

Summary The aim of this study was to investigate the effects of a gelatin-based plasma expander on blood coagulation and haemostasis in human subjects. Six healthy men were studied in a randomised, controlled cross-over study to investigate the effects of a 60 min intravenous infusion of either 1 l gelatin-based plasma substitute (Gelofusine) or 0.9% NaCl (control). The infusion of gelatin resulted in a 1.7 fold increase in bleeding time at 60 min and a 1.4 fold increase at 120 min, while saline had no effect (p <0.05). Aggregation studies revealed a significant impairment of ristocetin-induced platelet aggregation (p <0.05), associated with a substantial decrease of vWF:ag (–32% vs. –5%, p <0.05) and ristocetin co-factor (–29% vs. +1%, p <0.05) and without in vitro impairment of the platelet glycoprotein 1b receptor. Gelatin caused a decrease in thrombin-antithrombin complexes (–45% vs. –4%, p <0.05) and F1+2 (–40% vs. +1%, p <0.05). The decrease in circulating levels of vWF:ag, vWF R:Co, thrombin-antithrombin complexes and F1+ 2 was more than could be expected by the calculated plasma-dilution generated by Gelofusine. Our results demonstrated that the administration of a gelatin-based plasma substitute results in a significant impairment of primary haemostasis and thrombin generation. The defect in primary haemostasis appears to be related to a gelatin-induced reduction in von Willebrand factor, whereas the decreased thrombin generation may be due to the dilution of coagulation factors induced by Gelofusine.
Assessment of Sleep Complaints and Sleep-Disordered Breathing in a Consecutive Series of Obese Patients

Background/Objective: The prevalence of sleep-related complaints (SRC) and the frequency of sleep-disordered breathing (SDB) in obese patients has not been studied extensively. We investigated SRC and SDB in a group of obese persons as part of a preoperative workup for weight reduction (bariatric) surgery. Methods: All consecutive patients attending a weight-loss clinic for evaluation for bariatric surgery were asked to complete a questionnaire. The questionnaire consisted of a section on SRC and a validated general sleep questionnaire (Sleep Wake Experience List). The patients underwent sleep studies in which an Edentrace recorder registered heart rate, chest wall movements by impedance, airflow and oxygen saturation. Results: Fifty-one patients (14 men, 37 women) were evaluated. Mean body mass index (BMI) was 45 kg/m2 (range 33–61). Eighteen patients (35%) demonstrated SDB, defined as (a) an apnea/hypopnea index ≥5, and/or (b) more than 2% of registration time with an oxygen saturation below 90%. There was no difference between these 18 patients and patients who did not exhibit SDB in age, sex, BMI or SRC. Seven patients had SDB of a severity warranting closer investigation and perioperative monitoring. Conclusion: Both SRC and SDB are common in obese patients. Limited nocturnal respiratory monitoring is indicated as part of the preoperative workup for weight reduction surgery.
Comparative study of the sensitivity of different diagnostic methods for the laboratory diagnosis of Buruli ulcer disease.

BACKGROUND Several diagnostic laboratory methods are available for case confirmation of Buruli ulcer disease. This study assessed the sensitivity of various diagnostic tests in relation to clinical presentation of the disease, type of diagnostic specimen, and treatment history. METHODS Swab samples, 3-mm punch biopsy tissue specimens, and surgically excised tissue specimens from 384 individuals with suspected Buruli ulcer disease were obtained at 9 different study sites in Ghana and were evaluated with dry reagent-based polymerase chain reaction (PCR), microscopic examination, culture, and histopathological analysis. The study subjects presented with nonulcerative and ulcerative lesions and were divided into 3 treatment groups: (1) previously untreated patients scheduled for antimycobacterial treatment, (2) patients treated with surgery alone, and (3) patients treated with surgery in combination with previous antimycobacterial treatment. RESULTS Of 384 suspected cases of Buruli ulcer disease, 268 were confirmed by at least 1 positive test result. The overall sensitivity of PCR (85%) was significantly higher than that of microscopic examination (57%) and culture (51%). After data were stratified by treatment group, type of lesion, and diagnostic specimen type, analysis revealed that PCR of 3-mm punch biopsy tissue specimens (obtained from previously untreated nonulcerative lesions) and of swab samples (obtained from previously untreated ulcers) had the highest diagnostic sensitivity (94% and 90%, respectively). Although duration of the disease did not significantly influence the sensitivity of any test, previous antimycobacterial treatment was significantly associated with decreased sensitivity of PCR and culture. CONCLUSIONS Across all subgroups, PCR had the highest sensitivity. PCR assessment of 3-mm punch biopsy tissue specimens proved to be the best diagnostic tool for nonulcerative lesions, and PCR assessment of swab samples was the best diagnostic tool for ulcerative lesions. For monitoring of antimycobacterial treatment success within controlled trials, however, only culture is appropriate.
Calcium spike variability in cardiac myocytes results from activation of small cohorts of ryanodine receptor 2 channels

• Cardiac myocytes contract due to the upsurge in intracellular calcium concentration resulting from the activation of ryanodine receptor (RyR) channels residing at calcium releasing sites. • Calcium release can be observed as local calcium spikes that vary in amplitude, time course and frequency of occurrence due to reasons that are not well understood. • In this study we quantify the characteristics of calcium spikes in isolated myocytes, typify them by stochastic analysis and interpret them by mathematical modelling. • We show that the observable properties of calcium spikes are compatible with gating properties, calcium/magnesium sensitivity and distribution of RyR channels, if only a few out of the many RyRs clustered at the release sites activate at a single release event. • The proposed mechanism of stimulated calcium release clarifies the concepts of the structure and function of calcium releasing sites and of plasticity and dynamics of calcium signalling in cardiac development and disease.
Prognosis of Critical Limb Ischemia in Hemodialysis Patients After Isolated Infrapopliteal Balloon Angioplasty: Results From the Japan Below-the-Knee Artery Treatment (J-BEAT) Registry

Purpose To evaluate the long-term clinical results after isolated infrapopliteal balloon angioplasty for critical limb ischemia (CLI) in end-stage renal disease patients on hemodialysis. Methods Between April 2004 and October 2010, 406 CLI consecutive patients (275 men; mean age 71±11 years) who underwent balloon angioplasty for primary treatment of isolated infrapopliteal artery lesions in 465 limbs were enrolled in a multicenter, nonrandomized registry. The patients were classified into 2 groups, those on hemodialysis (242 patients with 283 limbs) and those not (164 patients with 182 limbs), for a retrospective comparative study of clinical outcomes [target extremity revascularization (TER), major amputation (MA), and survival] at an average 3.4±1.9 years. Results Freedom from TER at 5 years was lower in the hemodialysis patients (48.3% vs. 65.4% in non-hemodialysis patients, p<0.001); 9.9% of hemodialysis patients had undergone bypass surgery in contrast to 3.8% of non-hemodialysis patients (p=0.011). Freedom from MA was 77.1% in hemodialysis patients and 85.1% in non-hemodialysis patients at 5 years (p=0.058). Hemodialysis patients had significantly poorer survival (24.3% vs. 48.0%, p<0.001) and MA-free survival (20.8% vs. 42.9%, p<0.001) than non-hemodialysis patients at 5 years. Multivariate predictors of MA or all-cause death were non-ambulatory status (p<0.001), gangrene (p=0.036), and higher C-reactive protein levels (p=0.048). Conclusion Although hemodialysis patients have a higher TER rate compared to the general population, the long-term limb salvage rate after balloon angioplasty for isolated infrapopliteal lesions is acceptable; nevertheless, the MA-free survival rate is very low.
Identification and purification of proteins from germ cell‐conditioned medium (GCCM)

Germ cells are known to regulate Sertoli cell and testicutar function possibly through released factor(s) or via cell‐cell contact. However, the identities of many of these putative biological factors are not known. The aim of this study is to present a strategy to identify and purify germ cell‐derived proteins found in germ cell‐conditioned medium (GCCM) at a quantity sufficient to permit protein microsequencing. The purification scheme of a novel germ cell‐derived protein from GCCM designated GC‐26 is presented along with several germ cell proteins using a combination of high pressure liquid chromatography (HPLC) columns. The purity of GC‐26 and other germ cell proteins were confirmed by sodium dodecyl sulfate (SDS)‐polyacrylamide gel electrophoresis (PAGE) and silver staining. The identities of GC‐26, a 26‐kDa polypeptide, and other proteins were determined by direct protein microsequencing. These partial NH2‐terminal amino acid sequences were compared with the existing databases at Protein Identification Resource (PIR), GenBank, and BLAST. These analyses revealed that these proteins are unique. This strategy should be useful for the micropurification of proteins from other biological samples and/or fluids.
Prevalence and correlates of depression among patients with head and neck cancer: a systematic review of implications for research.

PURPOSE/OBJECTIVES To present a comprehensive summary of the existing research literature related to prevalence and correlates of depression in adult patients with head and neck cancer to establish a knowledge base for future research. DATA SOURCES Quantitative studies in English measuring depression or mood in adults with head and neck cancer published from 1986-2008. DATA SYNTHESIS A substantial body of knowledge exists regarding prevalence, correlates, and predictors of depression in patients with head and neck cancer. Prevalence rates of depression are high at diagnosis, during treatment, and in the first six months following treatment, and mild to moderate depression may continue for three to six years after diagnosis. Certain patient demographic characteristics (e.g., marital status, education), symptoms, and specific time points in the illness trajectory (e.g., time of treatment) are correlated with depression. Specific patient variables at diagnosis, such as depression, can predict depression at later time points. CONCLUSIONS Additional research should assess symptoms using consistent depression instruments or clinical interviews based on specific criteria in patients with head and neck cancer. Specifically, multisite studies should be conducted to increase sample sizes. Research related to symptom clusters and the effect of clusters on patients is needed. Longitudinal studies that examine depression and patient characteristics, symptoms, type of treatments, and the correlates of depression across the trajectory of illness are important. Replication of existing research using multiple patient and clinical characteristics to explore predictors of depression may reveal profiles for patients most at risk. IMPLICATIONS FOR NURSING This comprehensive summary of existing research literature related to the prevalence and correlates of depression among adult patients with head and neck cancer provides evidence-based information that can be used by oncology nurses in their practice.
Somatic PIK3CA mutations in seven patients with PIK3CA‐related overgrowth spectrum

Somatic mutations in PIK3CA cause many overgrowth syndromes that have been recently coined the “PIK3CA‐Related Overgrowth Spectrum. ” Here, we present seven molecularly confirmed patients with PIK3CA‐Related Overgrowth Spectrum, including patients with Congenital Lipomatous Overgrowth, Vascular Malformations, Epidermal Nevi, Scoliosis/Skeletal and Spinal syndrome, Klippel–Trenaunay syndrome, lymphatic malformation and two with atypical phenotypes that cannot be classified into existing disease categories. The literature on PIK3CA‐Related Overgrowth Spectrum, suggests that PIK3CA c.1258T>C; p.(Cys420Arg), c.1624G>A; p.(Glu542Lys), c.1633G>A; p.(Glu545Lys), c.3140A>G; p.(His1047Arg), and c.3140A>T; p.(His1047Leu) can be identified in approximately 90% of patients without brain overgrowth. Therefore, droplet digital polymerase chain reaction targeting these mutation hotspots could be used as the first‐tier genetic test on patients with PIK3CA‐Related Overgrowth Spectrum who do not have signs of overgrowth in their central nervous system. © 2017 Wiley Periodicals, Inc.