Miracle-dz/UniFAR-FTUs huggingface dataset:
Maintenance of alpha-helical structures by phenyl rings in the active-site tyrosine triad contributes to catalysis and stability of ketosteroid isomerase from Pseudomonas putida biotype B.

Ketosteroid isomerase (KSI) from Pseudomonas putida biotype B is a homodimeric enzyme catalyzing an allylic rearrangement of Delta5-3-ketosteroids at rates comparable with the diffusion-controlled limit. The tyrosine triad (Tyr14.Tyr55.Tyr30) forming a hydrogen-bond network in the apolar active site of KSI has been characterized in an effort to identify the roles of the phenyl rings in catalysis, stability, and unfolding of the enzyme. The replacement of Tyr14, a catalytic residue, with serine resulted in a 33-fold decrease of kcat, while the replacements of Tyr30 and Tyr55 with serine decreased kcat by 4- and 51-fold, respectively. The large decrease of kcat for Y55S could be due to the structural perturbation of alpha-helix A3, which results in the reorientation of the active-site residues as judged by the crystal structure of Y55S determined at 2.2 A resolution. Consistent with the analysis of the Y55S crystal structure, the far-UV circular dichroism spectra of Y14S, Y30S, and Y55S indicated that the elimination of the phenyl ring of the tyrosine reduced significantly the content of alpha-helices. Urea-induced equilibrium unfolding experiments revealed that the DeltaG(U)H2O values of Y14S, Y30S, and Y55S were significantly decreased by 11.9, 13.7, and 9.5 kcal/mol, respectively, as compared with that of the wild type. A characterization of the unfolding kinetics based on PhiU-value analysis indicates that the interactions mediated by the tyrosine triad in the native state are very resistant to unfolding. Taken together, our results demonstrate that the internal packing by the phenyl rings in the active-site tyrosine triad contributes to the conformational stability and catalytic activity of KSI by maintaining the structural integrity of the alpha-helices.
Association of Asthma Diagnosis, Severity, Symptoms, and Treatment With Risk of Preeclampsia

OBJECTIVE: Existing studies relating asthma and preeclampsia provide conflicting results, perhaps due to differences in study populations, varying definitions of asthma, and inadequate control for confounding, particularly asthma medication use. This prospective study examines associations between aspects of asthma (diagnosis, severity, symptoms, and medication use) and risk of preeclampsia. METHODS: A total of 1,708 pregnant women, of whom 656 had asthma diagnosis and 1,052 had no asthma diagnosis, were included in this analysis. Asthma symptoms, treatment, and severity were classified according to Global Initiative for Asthma guidelines. Hospital records were abstracted, and strict criteria were applied to classify women as preeclamptic based on National Heart, Lung, and Blood Institute guidelines. RESULTS: There were 568 of 656 women with diagnosed asthma and 353 of 1,052 women without asthma diagnosis who had symptoms or took asthma medication during pregnancy. Separate adjusted logistic regression models were run for different measures of asthma status: 1) asthma diagnosis; 2) overall Global Initiative for Asthma severity; 3) Global Initiative for Asthma symptom and treatment steps; and 4) Global Initiative for Asthma symptom step and medication type. Women at increased risk of preeclampsia were those classified as Global Initiative for Asthma symptom step 3/4 compared with no symptoms (odds ratio 3.36, 95% confidence interval 1.24–9.14) and theophylline users (odds ratio 1.16 for every dose/month increase in use, 95% confidence interval 1.02–1.33). In contrast, neither a history of physician-diagnosed asthma nor Global Initiative for Asthma treatment step was associated with preeclampsia status. CONCLUSION: Our findings suggest that women with moderate to severe asthma symptoms, regardless of asthma diagnosis or treatment, are at increased risk of preeclampsia compared with women with no symptoms. LEVEL OF EVIDENCE: II-2
Housebound versus nonhousebound patients with myalgic encephalomyelitis and chronic fatigue syndrome

Objectives The objective of this study was to examine individuals with myalgic encephalomyelitis and chronic fatigue syndrome who are confined to their homes due to severe symptomatology. The existing literature fails to address differences between this group, and less severe, nonhousebound patient populations. Methods Participants completed the DePaul Symptom Questionnaire, a measure of myalgic encephalomyelitis and chronic fatigue syndrome symptomology, and the SF-36, a measure of health impact on physical/mental functioning. ANOVAs and, where appropriate, MANCOVAS were used to compare housebound and nonhousebound patients with myalgic encephalomyelitis and chronic fatigue syndrome across areas of functioning, symptomatology, and illness onset characteristics. Results Findings indicated that the housebound group represented one quarter of the sample, and were significantly more impaired with regards to physical functioning, bodily pain, vitality, social functioning, fatigue, postexertional malaise, sleep, pain, neurocognitive, autonomic, neuroendocrine, and immune functioning compared to individuals who were not housebound. Discussion Findings indicated that housebound patients have more impairment on functional and symptom outcomes compared to those who were not housebound. Understanding the differences between housebound and not housebound groups holds implications for physicians and researchers as they develop interventions intended for patients who are most severely affected by this chronic illness.
How do year one students represent and solve word problems based on numbers parts-whole relationship?

Numbers parts-whole relationship is mainly represented by number bonds in the Malaysian year one mathematics textbooks. Seldom tasks are given to encourage students to create their own representations. The purpose of this study was to investigate how year one students represent and solve word problems based on numbers parts-whole relationship. On this account a cross sectional survey was conducted to collect data from a purposive sample of 42 year one students chosen based on enrollment and their mathematics performance in LINUS. Their numbers parts-whole representation was examined with the `Number Pairs Instrument' and a short interview. Results indicated that though the students have been taught about using representations for finding number pairs, they faced difficulty in producing accurate and complete representations by themselves to solve the word problems. This study shows that year one students need guidance and practice for creating efficient representations to solve parts-whole word problems which require multiple answers.
MDM2 Inhibits PCAF (p300/CREB-binding Protein-associated Factor)-mediated p53 Acetylation*

Our previous study shows that MDM2, a negative feedback regulator of the tumor suppressor p53, inhibits p300-mediated p53 acetylation. Because PCAF (p300/CREB-binding protein-associated factor) also acetylates and activates p53 after DNA damage, in this study we have examined the effect of MDM2 on PCAF-mediated p53 acetylation. We have found that MDM2 inhibited p53 acetylation by PCAF in vitro. In addition, when overexpressed, MDM2 inhibited PCAF-mediated p53 acetylation in cells. MDM2 interacted with PCAF both in vitro and in cells, as assessed using GST fusion protein interaction and immunoprecipitation assays, respectively. Consistent with the above results, MDM2 significantly repressed the activation of p53 transcriptional activity by PCAF without apparently affecting the level of p53. In addition, MDM2 co-resided with p53 at the p53-responsivemdm2 and p21 waf1/cip1 promoters, inhibiting expression of the endogenous p21 waf1/cip1 . These results demonstrate that MDM2 can inhibit PCAF-mediated p53 acetylation and activation.
β-Arrestins and Cell Signaling

Upon their discovery, beta-arrestins 1 and 2 were named for their capacity to sterically hinder the G protein coupling of agonist-activated seven-transmembrane receptors, ultimately resulting in receptor desensitization. Surprisingly, recent evidence shows that beta-arrestins can also function to activate signaling cascades independently of G protein activation. By serving as multiprotein scaffolds, the beta-arrestins bring elements of specific signaling pathways into close proximity. beta-Arrestin regulation has been demonstrated for an ever-increasing number of signaling molecules, including the mitogen-activated protein kinases ERK, JNK, and p38 as well as Akt, PI3 kinase, and RhoA. In addition, investigators are discovering new roles for beta-arrestins in nuclear functions. Here, we review the signaling capacities of these versatile adapter molecules and discuss the possible implications for cellular processes such as chemotaxis and apoptosis.
Expression pattern of aquaporin 4 and 5 in the middle ear of guinea pigs with secretory otitis media

Conclusions: Our study suggests that aquaporins 4 and 5 (AQP4 and AQP5) in the middle ear cavity may play a vital role in the homeostasis of the tubotympanum and in the course of the accumulation of the effusion in secretory otitis media (SOM). Objective: To explore the pathological change in water homeostasis in the middle ear in SOM and to observe the expression and regulation of AQP4 and AQP5 in the middle ear cavity in SOM. Materials and methods: Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting were used to detect AQP4 and AQP5 in the bullae of animal models of SOM and normal animals. The expression patterns of AQP4 and AQP5 in the SOM group were compared with those in the normal group. Results: RT-PCR and immunoblot analyses revealed that mRNAs encoding AQP4 and AQP5 were expressed in the middle ear membrane of the guinea pigs in both groups; AQP4 was also detected as 33 kDa protein in both groups. Quantitative analysis of RT-PCR and Western blotting revealed that expression of AQP4 and AQP5 was higher in the SOM group than in the control group.
Growth and development of Strongyloides venezuelensis Brumpt, 1934 in the albino rat

Growth and development of S. venezuelensis in percutaneously infected albino rats was studied and compared with S. ratti. The development of S. venezuelensis was found to last longer than that of S. ratti, the difference resulting mainly from the longer sojourn of the L3 larvae in skin and lungs. The growth rate was found to differ in each developmental stage, being higher in L4 than in L3 larvae. The adult females continued to increase in size for 6–7 days postexposure. The increase in body width, oesophagus length and tail length was found to be allometric bradyauxetic.
Vasopressin V2 receptor enhances gain of baroreflex in conscious spontaneously hypertensive rats.

The aim of the present study was to determine the receptor subtype involved in arginine vasopressin (AVP)-induced modulation of baroreflex function in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats using novel nonpeptide AVP V1- and V2-receptor antagonists. Baroreceptor heart rate (HR) reflex was investigated in both SHR and WKY rats which were intravenously administered the selective V1- and V2-receptor antagonists OPC-21268 and OPC-31260, respectively. Baroreflex function was assessed by obtaining alternate pressor and depressor responses to phenylephrine and sodium nitroprusside, respectively, to construct baroreflex curves. In both SHR and WKY rats baroreflex activity was tested before and after intravenous administration of vehicle (20% DMSO), OPC-21268 (10 mg/kg), and OPC-31260 (1 and 10 mg/kg). Vehicle did not significantly alter basal mean arterial pressure (MAP) and HR values or baroreflex function in SHR or WKY rats. The V1-receptor antagonist had no significant effect on resting MAP or HR values or on baroreflex parameters in both groups of rats, although this dose was shown to significantly inhibit the pressor response to AVP (5 ng iv; ANOVA, P < 0.05). In SHR but not WKY rats the V2-receptor antagonist significantly attenuated the gain (or slope) of the baroreflex curve (to 73 ± 3 and 79 ± 7% of control for 1 and 10 mg/kg, respectively), although AVP-induced pressor responses were also attenuated with the higher dose of the V2-receptor antagonist. These findings suggest that AVP tonically enhances baroreflex function through a V2 receptor in the SHR.
Fidelity to a behavioral intervention to improve goals of care decisions for nursing home residents with advanced dementia

Background/aims: Ensuring fidelity to a behavioral intervention implemented in nursing homes requires awareness of the unique considerations of this setting for research. The purpose of this article is to describe the goals of care cluster-randomized trial and the methods used to monitor and promote fidelity to a goals of care decision aid intervention delivered in nursing homes. Methods: The cluster randomized trial tested whether a decision aid for goals of care in advanced dementia could improve (1) the quality of communication and decision-making, (2) the quality of palliative care, and (3) the quality of dying for nursing home residents with advanced dementia. In 11 intervention nursing homes, family decision-makers for residents with advanced dementia received a two-component intervention: viewing a video decision aid about goals of care choices and then participating in a structured decision-making discussion with the nursing home care plan team, ideally within 3 months after the decision aid was viewed. Following guidelines from the National Institutes of Health Behavior Change Consortium, fidelity was assessed in study design, in nursing home staff training for intervention implementation, and in monitoring and receipt of the intervention. We also monitored the content and timing of goals of care discussions. Results: Investigators enrolled 151 family decision-maker/resident dyads in intervention sites; of those, 136 (90%) received both components of the intervention, and 92%–99% of discussions addressed each of four recommended content areas—health status, goals of care, choice of a goal, and treatment planning. A total of 94 (69%) of the discussions between family decision-makers and the nursing home care team were completed within 3 months. Conclusion: The methods we used for intervention fidelity allowed nursing home staff to implement a goals of care decision aid intervention for advanced dementia. Key supports for implementation included design features that aligned with nursing home practice, efficient staff training, and a structured guide for goals of care discussions between family decision-makers and staff. These approaches may be used to promote fidelity to behavioral interventions in future clinical trials.
Growth of Vibrio anguillarum in Salmon Intestinal Mucus

The physiological changes of Vibrio anguillarum in response to growth in salmon intestinal mucus were investigated. Growth, survival, and changes in protein expression during growth in media supplemented with mucus were compared to growth and starvation in the identical media without mucus. V. anguillarum exhibited a rapid decline in CFU following growth in mucus as the sole carbon source. No such decline was observed in Luria broth with a 2% NaCl concentration, in glucose-minimal broth (3M), or during starvation in a carbon-, nitrogen-, and phosphorus-free salt solution (NSS). The changes in protein expression during growth in mucus were examined by labeling cells with [(sup35)S]methionine and analyzing the labeled proteins by one- and two-dimensional gel electrophoresis and autoradiography. Comparison of [(sup35)S]methionine-labeled proteins from mucus-grown cells with 3M-grown cells and NSS-starved cells revealed four de novo mucus-inducible proteins (Mips). These Mips were localized in the membrane fraction of V. anguillarum. Additionally, at least one other membrane protein was found to have increased expression in response to growth in mucus.
ALLOZYME FREQUENCIES IN A LINEAR SERIES OF SONG DIALECT POPULATIONS

Evaluating the biological consequences of avian song dialects is important in at least two ways. First, where appropriate experimental evidence is available, we know that dialect markers in song are learned (Marler, 1970; Marler and Mundinger, 1971). Dialect systems thus offer an opportunity to determine if and how culturally transmitted vocal signals, a fundamental feature of many songbird societies, influence patterns of gene flow. Second, despite much fine descriptive work, few attempts have been made to connect the field of behavioral research on song dialects to that of evolutionary genetics. The central issue is whether or not population specific vocal signals (dialects) cause mating to take place primarily among members of the same dialect population. This question does not specify any particular behavioral mechanism that might be hypothesized nor should it suggest that gene migration is an all-or-none phenomenon. Gene flow between adjacent dialects may vary in amount or direction depending upon such possible influences as dialect discrimination affecting mate selection, recency of contact of the populations (Remington, 1968), availability of unoccupied breeding habitat, or major environmental disturbances. If song dialects reduce gene migration among dialect populations, however, then it is reasonable to predict that neighboring dialect populations may exhibit genetic differences (Templeton, 1980a). The divergence may occur because of founder effect, genetic drift, or because of selective differences in occupied habitats as when, for example, dialects occur along an environmental gradient (Endler, 1977). differential coadaptation of genomes may also be involved. Possible outcomes of the interaction of gene flow, environmental gradients, magnitude of selection pressures, and population size have only recently begun to yield to theoretical analysis (Endler, 1977; Slatkin, 1978; Lande, 1980) and we hope that our studies may be of some use in this regard. Our field data, while not constituting a test of this developing theory, may serve to underscore the significance of gene flow in population differentiation. The four song dialect populations of Whitecrowned Sparrows treated here extend over a total of only 35 km distance in an environment of apparent homogeneity in climate and vegetation. In view of the potentially high vagility of these birds, the possible consequences of discontinuities in gene flow caused by social structure may be more easily discovered. In this report we provide allozyme frequency data from four dialect populations occurring in a linear sequence along the central California coast. From these data we describe the nature and extent of genetic differentiation within and among the four song dialect populations to test the hypothesis of genetic differentiation of dialects.
A stable nanoparticulate DDA/MMG formulation acts synergistically with CpG ODN 1826 to enhance the CD4⁺ T-cell response.

AIM To combine the dimethyldioctadecyl ammonium/monomycoloyl glycerol (DDA/MMG) liposomal vaccine adjuvant with the Toll-like receptor (TLR) ligands poly(I:C) (TLR3), flagellin (TLR5) or CpG oligodeoxynucleotide 1826 (TLR9) and investigate their physicochemical properties as well as their CD4(+) T-cell-inducing capacity. MATERIALS & METHODS Formulations were investigated by dynamic light scattering and differential scanning calorimetry. Their CD4(+) T-cell induction with a tuberculosis antigen was analyzed by multiplex cytokine analysis, ELISA and intracellular cytokine staining. RESULTS DDA/MMG/CpG was the best combination for obtaining increased CD4(+) T-cell responses. However, coformulating CpG and DDA/MMG liposomes led to instability and the formulation was therefore optimized systematically using a design of experiment. CONCLUSION The nanoparticulate DDA/MMG/CpG adjuvant can be stabilized and synergistically enhances CD4(+) T-cell responses compared with DDA/MMG liposomes.
Decompression sickness Role of patent foramen ovale is limited

EDITOR, — Wilmshurst et al have reported an association between patent foramen ovale and some forms of decompression sickness.1 Our initial experience of performing contrast echocardiography in divers with decompression sickness, which we reported in a letter rather than a peer reviewed paper because we studied our subjects in an uncontrolled and non-blinded manner, failed to confirm this.2 J J W Sykes states that the link between a shunt and decompression sickness is controversial, giving a reference to our letter.3 Peter Wilmshurst, who criticises our findings,4 will recall that at the conclusion of …
Nature and mechanism of action of the CAMP protein of group B streptococci

The extracellular product of group B streptococci responsible for the CAMP reaction was purified to near homogeneity. It is a relatively thermostable protein having a molecular weight of 23,500 and an isoelectric pH of 8.3. It was found that the CAMP reaction could be simulated by substituting [14C]glucose-containing liposomes prepared from sphingomyelin, cholesterol, and dicetyl phosphate for sheep erythrocytes. In the belief that the liposome system is a valid model, the mechanism of the CAMP reaction was further investigated by using liposomes in which N-acylsphingosine (ceramide) was substituted for sphingomyelin. In this system disruption of liposomes, as measured by release of trapped [14C]glucose, was effected by CAMP protein alone. As judged from thin-layer chromatography, CAMP protein caused no reduction in the amount of ceramide present in ceramide-containing liposomes, nor were split products demonstrable. However, binding of CAMP protein to ceramide-containing liposomes could be shown. It is inferred that in sheep erythrocytes CAMP protein reacts nonenzymatically with membrane ceramide formed by the prior action of staphylococcal sphingomyelinase and that binding of CAMP protein to ceramide disorganizes the lipid bilayer to an extent that results in cell lysis.