Miracle-dz/UniFAR-FTUs huggingface dataset:
Ala Scanning of the Inhibitory Region of Cardiac Troponin I*

In skeletal and cardiac muscles, troponin (Tn), which resides on the thin filament, senses a change in intracellular Ca2+ concentration. Tn is composed of TnC, TnI, and TnT. Ca2+ binding to the regulatory domain of TnC removes the inhibitory effect by TnI on the contraction. The inhibitory region of cardiac TnI spans from residue 138 to 149. Upon Ca2+ activation, the inhibitory region is believed to be released from actin, thus triggering actin-activation of myosin ATPase. In this study, we created a series of Ala-substitution mutants of cTnI to delineate the functional contribution of each amino acid in the inhibitory region to myofilament regulation. We found that most of the point mutations in the inhibitory region reduced the ATPase activity in the presence of Ca2+, which suggests the same region also acts as an activator of the ATPase. The thin filaments can also be activated by strong myosin head (S1)-actin interactions. The binding of N-ethylmaleimide-treated myosin subfragment 1 (NEM-S1) to actin filaments mimics such strong interactions. Interestingly, in the absence of Ca2+ NEM-S1-induced activation of S1 ATPase was significantly less with the thin filaments containing TnI(T144A) than that with the wild-type TnI. However, in the presence of Ca2+, there was little difference in the activation of ATPase activity between these preparations.
Anti-activator ExsD Forms a 1:1 Complex with ExsA to Inhibit Transcription of Type III Secretion Operons*

The ExsA protein is a Pseudomonas aeruginosa transcriptional regulator of the AraC/XylS family that is responsible for activating the type III secretion system operons upon host cell contact. Its activity is known to be controlled in vivo through interaction with its negative regulator ExsD. Using a heterologous expression system, we demonstrated that ExsD is sufficient to inhibit the transcriptional activity of ExsA. Gel shift assays with ExsA- and ExsD-containing cytosolic extracts revealed that ExsD does not block DNA target sites but affects the DNA binding activity of the transcriptional activator. The ExsA-ExsD complex was purified after coproduction of the two partners in Escherichia coli. Size exclusion chromatography and ultracentrifugation analysis revealed a homogeneous complex with a 1:1 ratio. When in interaction with ExsD, ExsA is not able to bind to its specific target any longer, as evidenced by gel shift assays. Size exclusion chromatography further showed a partial dissociation of the complex in the presence of a specific DNA sequence. A model of the molecular inhibitory role of ExsD toward ExsA is proposed, in which, under noninducing conditions, the anti-activator ExsD sequesters ExsA and hinders its binding to DNA sites, preventing the transcription of type III secretion genes.
Social Lives of Medicines

Part I. Introduction: 1. An anthropology of materia medica Part II. The Consumers: 2. Mothers and children: the efficacies of drugs 3. Villagers and local remedies: the symbolic nature of medicines 4. Women in distress: medicines for control 5. Sceptical consumers: doubts about medicines Part III. The Providers: 6. Drug vendors and their market: the commodification of health 7. Pharmacists as doctors: bridging the sectors of health care 8. Injectionists: the attraction of technology 9. Prescribing physicians: medicines as communication Part IV. The Strategists: 10. Manufacturers: scientific claims, commercial aims 11. Health planners: making and contesting drug policy Part V. Conclusion: 12. Anthropologists and the sociality of medicines.
Molecular Determinants of the Interaction between Clostridium perfringens Enterotoxin Fragments and Claudin-3

Clostridium perfringens enterotoxin (CPE) binds to the extracellular loop 2 of a subset of claudins, e.g. claudin-3. Here, the molecular mechanism of the CPE-claudin interaction was analyzed. Using peptide arrays, recombinant CPE-(116–319) bound to loop 2 peptides of mouse claudin-3, -6, -7, -9, and -14 but not of 1, 2, 4, 5, 8, 10–13, 15, 16, 18–20, and 22. Substitution peptide mapping identified the central motif 148NPL150VP, supposed to represent a turn region in the loop 2, as essential for the interaction between CPE and murine claudin-3 peptides. CPE-binding assays with claudin-3 mutant-transfected HEK293 cells or lysates thereof demonstrated the involvement of Asn148 and Leu150 of full-length claudin-3 in the binding. CPE-(116–319) and CPE-(194–319) bound to HEK293 cells expressing claudin-3, whereas CPE-(116–319) bound to claudin-5-expressing HEK293 cells, also. This binding was inhibited by substitutions T151A and Q156E in claudin-5. In contrast, removal of the aromatic side chains in the loop 2 of claudin-3 and -5, involved in trans-interaction between claudins, increased the amount of CPE-(116–319) bound. These findings and molecular modeling indicate different molecular mechanisms of claudin-claudin trans-interaction and claudin-CPE interaction. Confocal microscopy showed that CPE-(116–319) and CPE-(194–319) bind to claudin-3 at the plasma membrane, outside cell-cell contacts. Together, these findings demonstrate that CPE binds to the hydrophobic turn and flanking polar residues in the loop 2 of claudin-3 outside tight junctions. The data can be used for the specific design of CPE-based modulators of tight junctions, to improve drug delivery, and as chemotherapeutics for tumors overexpressing claudins.
Incidence of heart failure in an urban population

The overall aim of the thesis was to study heart failure (HF) in an urban population, with reference to immigrant status, biological, life style and socioeconomic risk factors. The thesis is based on four papers. Paper I included inhabitants (N=114,917; aged 40-89 years) in the city of Malmo, Sweden, followed from November 1st, 1990 until December 31st, 2007. Paper II-IV used the population-based Malmo Diet and Cancer cohort (n=28,449; aged 45-73 years),followed from 1991-1996 until December 31st, 2008 or June 30th, 2009. Cases of HF were retrieved through the Swedish Hospital Discharge Register. Information on background characteristics in paper I was retrieved from the Population and Housing Census and the Swedish total population register. Participants in paper II-IV underwent sampling of peripheral venous blood, measurement of blood pressure and anthropometric measures and filled out a self-administered questionnaire. The risk of HF hospitalization was significantly higher among immigrants from Finland, Former Yugoslavia and Hungary compared to Swedish natives, after taking marital status, annual income and housing conditions into account. Furthermore, foreign-born had a significantly higher risk for HF hospitalization independently of hypertension, socioeconomic and several life-style risk factors. A significant interaction was seen between waist circumference and immigrant status on incident HF hospitalization; the increased HF risk was limited to immigrants with high waist circumference. Elevated body mass index, waist circumference, waist-hip ratio, body fat percentage and weight increased the risk of HF hospitalization in both sexes, independently of several sociodemographic, life style and biological factors. The joint exposure of high body mass index and high waist-hip ratio, or waist circumference, further increased the risk for HF hospitalization. In addition, the top quartile compared to the bottom quartile of red cell distribution width had a significantly higher risk for HF hospitalization after adjusting for other risk factors. In conclusion, there are substantial differences in risk of hospitalization due to HF among immigrants from different countries. Immigrant status was associated with risk of HF hospitalization independently of hypertension, socioeconomic and several life-style risk factors. Obesity is a risk factor for HF hospitalization, and the joint exposure to high body mass index and high waist-hip ratio or waist circumference further increased the HF risk. Red cell distribution width was found to be associated with long-term incidence of first hospitalization due to HF among middle-aged subjects. (Less)
Extra Corporeal Shock Wave Therapy Versus Local Corticosteroid Injection in the Treatment of Chronic Plantar Fasciitis, a Single Blinded Randomized Clinical Trial.

OBJECTIVES Plantar fasciitis is a self-limiting condition, but can be painful and disabling. Among the different treatments which exist, corticosteroid injections are effective and popular. Extracorporeal shock wave therapy (ESWT) is another treatment modality used for resistant conditions. In this study, the authors evaluated the efficacy of radial ESWT versus corticosteroid injections in the treatment of chronic plantar fasciitis. DESIGN Randomized clinical trial. SETTING Physical medicine and rehabilitation research center in a university hospital. SUBJECTS Forty patients with plantar fasciitis who did not respond to conservative treatment. METHODS Patients were allocated to radial ESWT with 2000 shock waves/session of 0.2 mJ/mm(2) (n = 20) or local methylprednisolone injections (n = 20). Pain in the morning and during the day based on a visual analog scale (VAS), functional abilities using the foot function index (FFI), and satisfaction were evaluated before treatment and at 4 and 8 weeks after treatment. RESULTS Patients (average age: 42.1± 8.20) received five sessions of ESWT or single steroid injection. Changes in the VAS in morning and during the day and the FFI throughout the study period were significant in both groups (P < 0.001). ESWT group had a higher reduction in VAS in morning and better function in FFI, but these changes were insignificant statistically [FFI decreased to 19.65 ± 21.26 points (67.4% improvement) in ESWT vs 31.50 ± 20.53 points (47.7%) in injection group at week 8, P = 0.072)]. Good or excellent results in the opinions of patients were achieved in 55% of ESWT and 30% of corticosteroid injection groups (P = 0.11). CONCLUSION Both interventions caused improvement in pain and functional ability 2 months after treatment. Although inter-group differences were not significant, the FFI was improved more with ESWT and patients were more satisfied with ESWT, thus shockwave therapy seems a safe alternative for management of chronic plantar fasciitis.
Identification and Functional Characterization of Protein Kinase A-catalyzed Phosphorylation of Potassium Channel Kv1.2 at Serine 449*

Vascular smooth muscle Kv1 delayed rectifier K+ channels (KDR) containing Kv1.2 control membrane potential and thereby regulate contractility. Vasodilatory agonists acting via protein kinase A (PKA) enhance vascule smooth muscle Kv1 activity, but the molecular basis of this regulation is uncertain. We characterized the role of a C-terminal phosphorylation site, Ser-449, in Kv1.2 expressed in HEK 293 cells by biochemical and electrophysiological methods. We found that 1) in vitro phosphorylation of Kv1.2 occurred exclusively at serine residues, 2) one major phosphopeptide that co-migrated with 449pSASTISK was generated by proteolysis of in vitro phosphorylated Kv1.2, 3) the peptide 445KKSRSASTISK exhibited stoichiometric phosphorylation by PKA in vitro, 4) matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectroscopy (MS) and MS/MS confirmed in vitro Ser-449 phosphorylation by PKA, 5) in situ phosphorylation at Ser-449 was detected in HEK 293 cells by MALDI-TOF MS followed by MS/MS. MIDAS (multiple reaction monitoring-initiated detection and sequencing) analysis revealed additional phosphorylated residues, Ser-440 and Ser-441, 6) in vitro 32P incorporation was significantly reduced in Kv1.2-S449A, Kv1.2-S449D, and Kv1.2-S440A/S441A/S449A mutant channels, but Kv1.2-S440A/S441A was identical to wild-type Kv1.2 (Kv1.2-WT), and 7) bath applied 8-Br-cAMP or dialysis with PKA catalytic subunit (cPKA) increased Kv1.2-WT but not Kv1.2-S449A current amplitude. cPKA increased Kv1.2-WT current in inside-out patches. Rp-CPT-cAMPS reduced Kv1.2-WT current, blocked the increase due to 8-Br-cAMP, but had no effect on Kv1.2-S449A. cPKA increased current due to double mutant Kv1.2-S440A/S441A but had no effect on Kv1.2-S449D or Kv1.2-S440A/S441A/S449A. We conclude that Ser-449 in Kv1.2 is a site of PKA phosphorylation and a potential molecular mechanism for Kv1-containing KDR channel modulation by agonists via PKA activation.
Reliability and Construct Validity of the Patient-Reported Outcomes Measurement Information System (PROMIS) Instruments in Women with Fibromyalgia

Objective The Patient-Reported Outcomes Measurement Information System (PROMIS) was developed to standardize measurement of clinically relevant patient-reported outcomes. This study evaluated the reliability and construct validity of select PROMIS static short-form (SF) instruments in women with fibromyalgia. Design Analysis of baseline data from the Fibromyalgia Activity Study with TENS (FAST), a randomized controlled trial of the efficacy of transcutaneous electrical nerve stimulation. Setting Dual site, university-based outpatient clinics. Subjects Women aged 20 to 67 years diagnosed with fibromyalgia. Methods Participants completed the Revised Fibromyalgia Impact Questionnaire (FIQR) and 10 PROMIS static SF instruments. Internal consistency was calculated using Cronbach alpha. Convergent validity was examined against the FIQR using Pearson correlation and multiple regression analysis. Results PROMIS static SF instruments had fair to high internal consistency (Cronbach α = 0.58 to 0.94, P < 0.05). PROMIS 'physical function' domain score was highly correlated with FIQR 'function' score (r = -0.73). The PROMIS 'total' score was highly correlated with the FIQR total score (r = -0.72). Correlations with FIQR total score of each of the three PROMIS domain scores were r = -0.65 for 'physical function,' r = -0.63 for 'global,' and r = -0.57 for 'symptom' domain. PROMIS 'physical function,' 'global,' and 'symptom' scores explained 58% of the FIQR total score variance. Conclusions Select PROMIS static SF instruments demonstrate convergent validity with the FIQR, a legacy measure of fibromyalgia disease severity. These results highlight the potential utility of select PROMIS static SFs for assessment and tracking of patient-reported outcomes in fibromyalgia.
Therapeutic Monoclonal Antibodies: Past, Present, and Future

In this chapter, an overview of the therapeutic antibody industry today, including the many commercial antibodies and Fc fusions and the rich clinical pipeline, is presented and analyzed. The long history of antibodies is given to bring context to the therapeutic antibody industry. This history includes serum therapy, the use of IVIG, and the evolution of those therapies into the development of the monoclonal Therapeutic Monoclonal Antibodies: From Bench to Clinic. Edited by Zhiqiang An Copyright # 2009 John Wiley & Sons, Inc. 3 antibody business as we know it today. The history of technologies that fostered the revolution of therapeutic antibody development in the 1990s is also described. Finally, the future of the therapeutic monoclonal antibody and Fc fusion business is presented along with opportunities and challenges facing the business and those who work in it. 1.
Comorbidity and Unmet Service Needs Among Medicaid-Enrolled Children With Identified Disabilities

The comorbidity of health and mental health problems among Medicaid-enrolled children and youth in Florida with a designated health or mental health disability was examined and compared to that of Medicaid-enrolled children with no disability. Slightly higher rates of health problems (42%) were found among children with mental health disabilities compared to the rate of mental health problems (35%) found among children with health disabilities.Within both groups, similar rates (~ 26%) of current comorbid health and mental health problems were found. Regarding health services, less than 1% to 11% of the children experiencing significant health problems were receiving no services, depending on disability group. Rates of unmet mental health needs were higher, ranging from 4% among children with no identified disability to 22% among children with an identified health disability.These data support the need for a comprehensive health policy emphasizing the assessment of all Medicaid-enrolled children with disabilities for comorbid health or mental health problems.
Ethical Dilemmas of Practicing Social Workers Around Psychiatric Medication

Abstract It is acknowledged that social workers in mental health and other settings routinely experience client-related ethical dilemmas. Further, there is wide recognition of the potential impact of ethical dilemmas on social work practice with clients who use psychotropic medication. Little is know empirically, however, about the experiences of practitioners with these dilemmas. This article describes the results of a national survey of practicing social workers regarding the nature of ethical dilemmas they face related to their work with clients on medication issues. The results make it clear that social workers regularly confront a variety of ethical dilemmas in this type of practice. Many of these dilemmas are related to ambiguities around the knowledge base of practice, appropriate roles of providers, and basic personal and professional values. The authors present implications of these findings for social work practice and further research.
Clinical information success in a maternity hospital

Sandra O'Connor (University Regional & Maternity Hospitals, Limerick, Ireland) Ita Richardson (Lero – the Irish Software Engineering Research Centre, University of Limerick) Jennifer Hogan (University Regional & Maternity Hospitals, Limerick, Ireland) Theresa O'Donnoghue (University Regional & Maternity Hospitals, Limerick, Ireland) Roy Philip (University Regional & Maternity Hospitals, Limerick, Ireland) Gerard Burke (University Regional & Maternity Hospitals, Limerick, Ireland) Abstract
Red blood cell transfusion need at diagnosis adversely affects survival in primary myelofibrosis—increased serum ferritin or transfusion load does not

Serum ferritin level at diagnosis was available in 185 patients with primary myelofibrosis (PMF); twenty‐two (12%) patients had serum ferritin >1,000 ng/mL and 32 (17%) were red blood cell (RBC) transfusion‐dependent. As expected, RBC transfusion need and increased serum ferritin displayed strong correlation (P < 0.0001); in addition, the latter but not the former correlated with advanced age (P < 0.0001). During median follow‐up of 28 months (range 0.5–231), peak serum ferritin levels exceeded 1,000 ng/mL in 41 (22%) patients. On multivariable analysis that included age as a covariate, RBC transfusion need at diagnosis (P < 0.0001), but not increased serum ferritin or transfusion load, predicted shortened survival. The prognostic relevance of RBC transfusion need was independent of the International Prognostic Scoring System and was also illustrated for leukemia‐free survival (P = 0.003). In PMF, the presence of a more severe erythropoietic defect, and not iron overload, has additional adverse prognostic value. Am. J. Hematol. , 2009. © 2009 Wiley‐Liss, Inc.
Genome Mining in Sorangium cellulosum So ce56

Myxobacteria, especially members of the genus Sorangium, are known for their biotechnological potential as producers of pharmaceutically valuable secondary metabolites. The biosynthesis of several of those myxobacterial compounds includes cytochrome P450 activity. Although class I cytochrome P450 enzymes occur wide-spread in bacteria and rely on ferredoxins and ferredoxin reductases as essential electron mediators, the study of these proteins is often neglected. Therefore, we decided to search in the Sorangium cellulosum So ce56 genome for putative interaction partners of cytochromes P450. In this work we report the investigation of eight myxobacterial ferredoxins and two ferredoxin reductases with respect to their activity in cytochrome P450 systems. Intriguingly, we found not only one, but two ferredoxins whose ability to sustain an endogenous So ce56 cytochrome P450 was demonstrated by CYP260A1-dependent conversion of nootkatone. Moreover, we could demonstrate that the two ferredoxins were able to receive electrons from both ferredoxin reductases. These findings indicate that S. cellulosum can alternate between different electron transport pathways to sustain cytochrome P450 activity.
Effects of Tendon Splitting on Experimentally-Induced Acute Equine Tendinitis

Summary The objective of this study was to compare the early phases of healing between surgically split and unsplit (control) tendons in a superficial digital flexor tendinitis model. Tendinitis was induced in both forelimbs of six horses by injection with collagenase. One tendon from each horse was randomly chosen to undergo tendon splitting. In six horses, the tendons were evaluated by ultrasonography before (week 0) and after (week 1) the onset of tendinitis and at weeks two, three, and four following tendon splitting. Three horses were euthanatized at the end of week four. Three horses were maintained for an additional month and evaluated at week eight before euthanasia. Following euthanasia, microangiographic and histologic evaluations were performed. Ultrasonographic evaluation revealed a significant (p <0.05) decrease in mean tendon lesion area in the split tendons when compared to the controls at weeks three, four, and eight. By week eight, all lesions had resolved in the split but not the control tendons. Mean lesion grade was lower in the split tendons at weeks two, three, four, and eight. Tendon area was similar (p >0.05) between groups at all periods after surgery. There was a more mature vascular pattern and greater peritendinous vascular response in the split tendons compared to control tendons. Histological examination under plain and polarized light revealed more normal collagen orientation and wave formation in the repair tissue in the split tendons at weeks four and eight. The fibroblast nuclei density was significantly (p <0.05) lower in the split tendons at week eight. These findings suggest that during the time period of this study, tendon splitting results in a more rapid decrease in lesion size and superior repair tissue organization when compared to controls. In a collagenase-induced tendinitis model, tendon splitting was shown to result in a greater reduction in tendon lesion size and superior repair tissue organization when compared to control tendons.
Modulation of M2‐type pyruvate kinase activity by the cytoplasmic PML tumor suppressor protein

The promyelocytic leukemia (PML) tumor suppressor protein accumulates in PML nuclear bodies (PML‐NBs), and can induce growth arrest, cellular senescence and apoptosis. PML has also been localized in the cytoplasm, although its function in this localization remains elusive. A general property of primary cancers is their high glycolytic rate which results from increased glucose consumption. However, the mechanism by which cancer cells up‐regulate glycolysis is not well understood. Here, we have shown that cytoplasmic PML (cPML) directly interacts with M2‐type pyruvate kinase (PKM2), a key regulator of carbon fate. PKM2 determines the proportion of carbons derived from glucose that are used for glycolytic energy production. Over‐expression of PML‐2KA mutant in the cytoplasm, which was generated by mutagenesis of the nuclear localization signals of PML, in MCF‐7 breast cancer cells suppressed PKM2 activity and the accumulation of lactate. PKM2 exists in either an active tetrameric form which has high affinity for its substrate phosphoenolpyruvate (PEP) or a less active dimeric form which has low affinity for its substrate. Over‐expression of PML‐2KA suppressed the activity of the tetrameric form of PKM2, but not the dimeric form. Our findings suggest that cPML plays a role in tumor metabolism through its interaction with PKM2.