Miracle-dz/UniFAR-FTUs huggingface dataset:
Anti-Human Immunodeficiency Virus Type 1 Activity of Novel 6-Substituted 1-Benzyl-3-(3,5-Dimethylbenzyl)Uracil Derivatives

ABSTRACT Nonnucleoside reverse transcriptase (RT) inhibitors (NNRTIs) are important components of current combination therapies for human immunodeficiency virus type 1 (HIV-1) infection. In screening of chemical libraries, we found 6-azido-1-benzyl-3-(3,5-dimethylbenzyl)uracil (AzBBU) and 6-amino-1-benzyl-3-(3,5-dimethylbenzyl)uracil (AmBBU) to be highly active and selective inhibitors of HIV-1 replication in vitro. To determine the resistance profiles of these compounds, we conducted a long-term culture of HIV-1-infected MT-4 cells with escalating concentrations of each compound. After serial passages of the infected cells, escape viruses were obtained, and they were more than 500-fold resistant to the uracil derivatives compared to the wild type. Sequence analysis was conducted for RT of the escape viruses at passages 12 and 24. The amino acid mutation Y181C in the polymerase domain of RT was detected for all escape viruses. Docking studies using the crystal structure of RT showed that AmBBU requires the amino acid residues Leu100, Val106, Tyr181, and Trp229 for exerting its inhibitory effect on HIV-1. Four additional amino acid changes (K451R, R461K, T468P, and D471N) were identified in the RNase H domain of RT; however, their precise role in the acquisition of resistance is still unclear. In conclusion, the initial mutation Y181C seems sufficient for the acquisition of resistance to the uracil derivatives AzBBU and AmBBU. Further studies are required to determine the precise role of each mutation in the acquisition of HIV-1 resistance.
Identification of Intron and Exon Sequences Involved in Alternative Splicing of Insulin Receptor Pre-mRNA*

The insulin receptor exists as two isoforms, A and B, that result from alternative splicing of exon 11 in the primary transcript. We have shown previously that the alternative splicing is developmentally and hormonally regulated. Consequently, these studies were instigated to identify sequences within the primary RNA transcript that regulate the alternative splicing. Minigenes containing exons 10, 11, and 12 and the intervening introns were constructed and transfected into HepG2 cells, which contain both isoforms of the insulin receptor. The cells were able to splice the minigene transcript to give both A (− exon 11) and B-like (+ exon 11) RNAs. A series of internal deletions within intron 10 were tested for their ability to give A and B RNAs. Intron 10 contained two sequences that modulated exon 11 inclusion; a 48-nucleotide purine-rich sequence at the 5′ end of intron 10 that functions as a splicing enhancer and causes an increase in exon 11 inclusion, and a 43-nucleotide sequence at the 3′ end of intron 10 upstream of the branch point sequence that favors skipping of exon 11. Increasing the length of the polypyrimidine tract at the 3′ end of intron 10 caused exon 11 to be spliced constitutively, indicating that a weak splice site is required for alternative splicing. Finally, point mutations, insertions, and deletions within exon 11 itself were able to regulate inclusion of the exon both positively and negatively.
Implementation of an inpatient smoking cessation programme in a Veterans Affairs facility.

AIMS AND OBJECTIVES To test the transportability and implementation of the Tobacco Tactics intervention using the Reach, Effectiveness, Adoption, Implementation and Maintenance framework, for inpatient units at the Jesse Brown Veterans Affairs Medical Center. BACKGROUND Smoking rates are high among veterans. While the Department of Veterans Affairs has standardised outpatient cessation clinics, inpatient cessation services, known to be efficacious, are only sporadically provided. DESIGN This was a phase 4, pre and postimplementation study of the Tobacco Tactics intervention. METHODS A unique convenience sample of inpatient veteran smokers was recruited both before (n = 54) and after (n = 50) implementation of the Tobacco Tactics programme. Participants completed baseline and 30-day follow-up surveys along with urine cotinine test kits. In addition, staff completed anonymous surveys during the preintervention period (n = 158) and two months after (n = 81) the Tobacco Tactics training. Bivariate analyses compared preintervention vs. postintervention patient and staff characteristics using Chi-square, Fisher's Exact or Student's t-test. p-values <0·05 were considered significant. RESULTS Patient-reported receipt of services and satisfaction was 10% higher in the postintervention compared to the preintervention group. Quit rates were 3% higher in the postintervention than in the preintervention group. The mean number of cigarettes smoked per day increased from 13 to 15 in the preintervention group, while the mean number of cigarettes smoked per day decreased from 14 to 9 in the postintervention group. Staff's confidence in their ability to provide cessation services improved greatly posttraining (p = 0·0017) as did self-reported delivery of cessation services (p = 0·0154). CONCLUSIONS With as little as one-hour training for nurses, the Tobacco Tactics intervention has the potential to be widely disseminated in the Department of Veterans Affairs. RELEVANCE TO CLINICAL PRACTICE The implementation of inpatient smoking interventions has the potential to improve quit rates and decrease morbidity and mortality in the Department of Veterans Affairs.
Nitric oxide synthase in human and rat lung: immunocytochemical and histochemical localization.

Nitric oxide synthase (NOS) produces nitric oxide, a mediator of potential importance in numerous physiologic and inflammatory processes in the lung. We localized constitutive NOS (c-NOS) and inducible NOS (i-NOS) within lung tissue by immunoperoxidase labeling with specific antibodies or by histochemical demonstration of the characteristic NADPH diaphorase activity of NOS. We analyzed human airway (n = 4) or parenchyma (n = 10) specimens obtained from uninvolved areas of surgical tumor resections. We also studied human fetal lung samples (n = 6) and normal or inflamed (16 h after intratracheal LPS instillation) rat lung tissue. Immunostaining with anti-c-NOS identified c-NOS antigen in rat lung nerves, endothelium, and airway epithelium. Normal or inflamed rat macrophages were not stained. Human nerve elements and large-vessel endothelium showed immunostaining with the anti-c-NOS, but no labeling of the airway or alveolar epithelium was seen. Immunostaining with anti-i-NOS showed strong labeling of rat macrophages after LPS treatment, in vivo or in vitro, while normals were negative. Human alveolar macrophages were occasionally positive for i-NOS, especially in areas of chronic inflammation, which also showed focal immunolabeling of endothelium. Uniform labeling of epithelium in large, cartilaginous airways was found with anti-i-NOS in both human bronchi and normal rat trachea samples, suggesting a constitutive role for a NOS that shares epitope(s) with or is highly homologous to the inducible, macrophage type of NOS. Histochemical staining for NADPH diaphorase activity was consistent with immunolocalization of NOS antigens. (ABSTRACT TRUNCATED AT 250 WORDS)
CD56 and PGP expression in acute myeloid leukemia: impact on clinical outcome.

BACKGROUND AND OBJECTIVES Overexpression of P-glycoprotein (PGP), a multidrug-related (MDR) protein, is one of the most important factors responsible for reduced drug sensitivity in acute myeloid leukemia (AML). Recently, we demonstrated that the presence of CD56 antigen, an isoform of the neural adhesion molecule, in AML cells is a negative independent prognostic factor for the achievement of complete remission (CR) and correlates with shorter survival. Since in our previous report we observed a more frequent PGP expression in CD56+ patients, we hypothesized that the reduced response to chemotherapy in this group of patients was due to increased PGP-mediated drug efflux. To confirm this hypothesis in this study PGP and CD56 expression on AML cells was correlated with other clinical and biological features and treatment response. DESIGN AND METHODS Immunophenotypic analysis, including evaluation of CD56 and PGP expression, was performed using multiparameter flow cytometry on fresh and/or cryopreserved blast cells, obtained after informed consent, from bone marrow and/or peripheral blood of 143 consecutive newly diagnosed AML cases at the time of diagnosis. Samples expressing CD56 in at least 15% or more cells were considered as positive (CD56+). PGP expression was expressed as a mean fluorescence index (MFI) i.e. as the ratio of sample mean fluorescence channel and the isotypic control mean fluorescence channel. RESULTS Overall results showed that 67/143 cases were PGP-/CD56-, 23/143 were PGP+ /CD56+, 40/143 were PGP+/CD56- and the remaining 13/143 were PGP-/CD56+. CD56+ and PGP+ on AML cells significantly reduced the CR rate (83% in the PGP-/CD56- group vs 60% in the PGP-/CD56+ group, 46% in the PGP+/CD56- group and 58% in the PGP+/CD56+ group, p = 0.002). In addition we observed a significantly higher proportion of total failures in patients expressing PGP or CD56 compared to in the group not expressing either (73% vs 27%, respectively; p = 0.0001). CD56 and PGP overexpression influenced the overall survival: in fact, the median survival of CD56+ and PGP+ patients ranged from 10 to 23 months, while the actuarial survival of CD56-/PGP- patients at 5 years is 52% (p = 0.023). INTERPRETATION AND CONCLUSIONS Our data underline the independent negative prognostic role of PGP and CD56 expression in acute myeloid leukemia. Since the mechanism by which CD56 reduces drug sensitivity is still unknown, further investigations are required.
A Phase 2 Study of Moderate Dose Interleukin-2 and Granulocyte-Macrophage Colony-Stimulating Factor in Patients With Metastatic or Unresectable Renal Cell Carcinoma

Interleukin-2 (IL-2) has been shown to produce durable complete remission in patients with renal cell carcinoma (RCC). A phase 2 study was conducted to evaluate the potential therapeutic synergy as well as the toxic side effects of the concurrent administration of IL-2 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in patients with advanced stage disease. Twenty-one patients with unresectable or metastatic RCC having an Eastern Oncology Cooperative Group performance status of 0 or 1 were enrolled. Six patients had received prior immunotherapy with interferon (IFN)-α, IFN-γ, and IL-12, whereas the remaining 15 subjects were previously untreated. Thirteen patients were assigned to a moderate-dose bolus of IL-2 at 72,000 IU/kg every 8 hours on days 1 through 5 and days 15 through 19, whereas 8 patients were given IL-2 as an intravenous continuous infusion at a dose of 5 MU/m2/d on days 1 through 5 and days 15 through 19. Subcutaneous GM-CSF at 125 μg/d on days 1 through 21 was administered concomitantly with IL-2. The median number of IL-2 bolus doses was 23 of a scheduled 28 (85%), whereas with the continuous infusion, 93% of planned IL-2 was given. All patients received 100% of GM-CSF doses. There were no complete or partial responses in this study. Of 13 patients treated in the bolus IL-2 arm, 10 had systemic progression of disease at 4 to 8 weeks, 1 developed metastasis in the brain at 4 weeks, and 2 had stable disease for 4 and 17 months. Among the 8 subjects treated with continuous infusion IL-2, 3 progressed with brain lesions at 3 to 8 weeks and 5 had stable disease at 6+, 7, 8+, 15+, and 17+ months. The median survival for the whole group was 10 months, with a range of 0.5 to 40+ months. There were no regimen-related deaths, and most of the observed toxicities were grade 1 and 2. Serious toxicities (grade 3 and 4) included anemia, atrial fibrillation, oliguria, abnormal liver function, and neurologic events like agitation or confusion. The combination of recombinant IL-2 and GM-CSF administered in the designed schedule and doses was not effective in patients with metastatic RCC and may even interfere with the therapeutic potential of moderate-dose IL-2 and increase its adverse events.
Changes in fetal prevalence and outcome for trisomies 13 and 18: a population-based study over 23 years

Objective. Changes in prenatal diagnosis and maternal age are likely to have an impact on live born prevalence of trisomies 13 and 18. We investigated trends in diagnosis, prevalence, and survival in these conditions. Methods. A population-based study of one UK health region in 1985–2007 using a well-established congenital abnormality register. Individual records were reviewed and live birth and maternal age data obtained. Results. Pregnancies with trisomies 13 and 18 increased from 0.08 to 0.23 per 1000 registered births and 0.20 to 0.65 per 1000 registered births, respectively. Prenatal diagnosis increased and was associated with high termination rates. Live born prevalence with trisomy 13 decreased from 0.05 to 0.03 per 1000 live births and with trisomy 18 from 0.16 to 0.10 per 1000 live births. Postnatal survival remains poor: one baby (3%) with trisomy 13 and four (6%) with trisomy 18 survived the first year. The percentage of mothers over 35 years increased from 6 to 15%. Conclusions. Changes in prenatal screening and maternal age have had dramatic effects on the live born prevalence of trisomies 13 and 18. Infant survival remains largely unchanged with the majority dying in the neonatal period.
Expression of the Na+/H+ exchanger regulatory protein family in genetically hypertensive rats

Objective To examine a possible involvement of a regulatory protein of Na+/H+ exchanger (NHE) in the increased renal NHE activity in spontaneously hypertensive rats (SHR), we investigated mRNA expression of inhibitory members of the NHE regulatory protein family, NHERF1 and NHERF2, in the kidney. Design Prehypertensive 4-week-old and hypertensive 11-week-old SHR and age-matched Wistar–Kyoto (WKY) rats were used to determine the changes in NHE activity and NHERF family expression in the kidney. Dahl salt sensitive (DS) and resistant rats were also used to examine whether these changes are specific for SHR. Methods mRNA expression in the kidney was quantified by RNase protection assay. The NHE activity in primary cultured proximal tubular cells was measured as Na-dependent pHi recovery rate by the NH4Cl prepulse technique with 2'7'-bis-(2-carboxyethyl)-5.6-carboxyfluorescein (BCECF). Results NHERF1 mRNA expression was significantly decreased in both prehypertensive and hypertensive SHR in comparison with age-matched WKY rats, whereas NHERF2 mRNA expression was significantly increased in SHR only in the hypertensive period. Antihypertensive treatment did not abolish these changes seen in control SHR. On the other hand, hypertensive DS rats fed a high-salt diet showed significant decreases in NHE activity and NHE3 mRNA expression compared with normotensive DS rats fed a low-salt diet, without significant changes in NHERF1 and NHERF2 mRNA expression. Conclusion These results suggest that decreased expression of NHERF1 may be related to the enhanced NHE activity in SHR and that these changes are likely to be genetically determined, whereas the increased NHERF2 expression may be induced as a compensatory mechanism.
Focal aggregation of voltage‐gated, Kv2.1 subunit‐containing, potassium channels at synaptic sites in rat spinal motoneurones

Delayed rectifier K+ currents are involved in the control of α‐motoneurone excitability, but the precise spatial distribution and organization of the membrane ion channels that contribute to these currents have not been defined. Voltage‐activated Kv2.1 channels have properties commensurate with a contribution to delayed rectifier currents and are expressed in neurones throughout the mammalian central nervous system. A specific antibody against Kv2.1 channel subunits was used to determine the surface distribution and clustering of Kv2.1 subunit‐containing channels in the cell membrane of α‐motoneurones and other spinal cord neurones. In α‐motoneurones, Kv2.1 immunoreactivity (‐IR) was abundant in the surface membrane of the soma and large proximal dendrites, and was present also in smaller diameter distal dendrites. Plasma membrane‐associated Kv2.1‐IR in α‐motoneurones was distributed in a mosaic of small irregularly shaped, and large disc‐like, clusters. However, only small to medium clusters of Kv2.1‐IR were observed in spinal interneurones and projection neurones, and some interneurones, including Renshaw cells, lacked demonstrable Kv2.1‐IR. In α‐motoneurones, dual immunostaining procedures revealed that the prominent disc‐like domains of Kv2.1‐IR are invariably apposed to presynaptic cholinergic C‐terminals. Further, Kv2.1‐IR colocalizes with immunoreactivity against postsynaptic muscarinic (m2) receptors at these locations. Ultrastructural examination confirmed the postsynaptic localization of Kv2.1‐IR at C‐terminal synapses, and revealed clusters of Kv2.1‐IR at a majority of S‐type, presumed excitatory, synapses. Kv2.1‐IR in α‐motoneurones is not directly associated with presumed inhibitory (F‐type) synapses, nor is it present in presynaptic structures apposed to the motoneurone. Occasionally, small patches of extrasynaptic Kv2.1‐IR labelling were observed in surface membrane apposed by glial processes. Voltage‐gated potassium channels responsible for the delayed rectifier current, including Kv2.1, are usually assigned roles in the repolarization of the action potential. However, the strategic localization of Kv2.1 subunit‐containing channels at specific postsynaptic sites suggests that this family of voltage‐activated K+ channels may have additional roles and/or regulatory components.
Drug use in primary open angle glaucoma: A prospective study at a tertiary care teaching hospital

Objective: To study drug use pattern in patients of primary open angle glaucoma (POAG) and to analyze the cost of different anti-glaucoma medications. Materials and Methods: This prospective study was carried in the glaucoma clinic of a tertiary care teaching hospital over a period of 9 months. The data collected for patients with POAG included the patient's demographic details and the drugs prescribed. Data were analyzed for drug use pattern and cost drugs used. Results: In a total 180 prescriptions (297 drugs) analyzed, most drugs (83.83%) were prescribed by topical route as eye drops. β blockers (93.88%) were found to be the most frequently prescribed for POAG. Timolol (82.22%) was the most frequently prescribed drug and timolol with acetazolamide (17.22%) was the most commonly prescribed drug combination. Fixed dose combinations constituted 26.66% of prescriptions. β blockers were found to be cheaper than other anti-glaucoma drugs while prostaglandins analogs were the costliest. Instructions about the route, frequency and duration of treatment were present in all prescriptions. However, instructions regarding instillation of eye drops were missing in all prescriptions.
The prognostic significance of pelvic lymph node morphology in carcinoma of the uterine cervix

Pelvic lymph nodes from 80 patients with primary invasive cervical cancer undergoing radical hysterectomy and pelvic lymphadenectomy were evaluated morphologically. Six to 25 (mean 14) lymph nodes from each patient were examined histologically and classified as follows: lymphocyte predominant, germinal center predominant, unstimulated, or lymphocyte depleted. Tumors were classified according to their primary cell type: large cell non‐keratinizing, keratinizing squamous cell, small cell, and adenocarcinoma. There was no direct relationship between lymph node morphology and patient age, lesion size, or tumor cell type. A lymphocyte predominant nodal pattern was associated with a statistically significant decrease in lymph node metastases, and tumor recurrence, and an increase in patient survival. In contrast, patients with a lymphocyte depleted nodal pattern had a high incidence of metastatic disease and tumor recurrence, and a decreased survival. Patients with lymph nodes showing an unstimulated or germinal center predominant pattern had an intermediate incidence of nodal metastases and tumor recurrence. These findings suggest that regional lymph nodal morphology may be of prognostic significance in patients with invasive carcinoma of the uterine cervix.
Promoting evidence-based practice: an organizational approach.

AIM To examine factors influencing the achievement of evidence-based practice. BACKGROUND This paper reports on the approach taken by a large teaching hospital in England to promote evidence-based practice. A summary of initiatives spanning a 5 year period is presented in order to set the scene for a recent survey of nurses to review progress made. Consideration is given to how the findings from the survey have been used to inform policy and practice. METHODS A survey by self-completed questionnaire was undertaken with a sample of clinical nurses (n = 330). The questionnaire examined the extent to which nurses utilised different sources of knowledge to inform their practice; perceived barriers to accessing evidence-based information and effecting change in practice; and a self-assessment of core skills necessary to underpin evidence-based practice. FINDINGS Nurses relied most heavily on experiential knowledge gained through their interactions with nursing colleagues, medical staff and patients to inform their practice. Organisational information in the form of policies and audit reports was drawn upon more frequently than research reports. Lack of time, resources and perceived authority to change practice influenced the extent to which nurses utilised formal sources of evidence. Whereas nurses were relatively well skilled at accessing and reviewing research evidence, they were less confident about their ability to change practice. CONCLUSION Health care organisations need to consider multiple strategies to facilitate and promote evidence-based practice. Managerial support, facilitation, and a culture that is receptive to change are essential.
Mediator complex association with constitutively transcribed genes in yeast

Mediator is a large, multisubunit complex that is essential for transcription of mRNA by RNA Pol II in eukaryotes and is believed to bridge transcriptional activators and the general transcription machinery. However, several recent studies suggest that the requirement for Mediator during transcriptional activation is not universal, but rather activator dependent, and may be indirect for some genes. Here we have investigated Mediator association with several constitutively transcribed genes in yeast by comparing a yeast strain that harbors a temperature-sensitive mutation in an essential Mediator subunit, Srb4, with its wild-type (WT) counterpart. We find modest association of Mediator with constitutively active genes and show that this association is strongly decreased in srb4 ts yeast, whereas association with a nontranscribed region or repressed gene promoters is lower and unaffected in the mutant yeast. The tail module of Mediator remains associated with ribosomal protein (RP) gene promoters in srb4 ts yeast, while subunits from the head and middle modules are lost. Tail module association at Rap1-dependent gene promoters is lost in rap1 ts yeast, indicating that Rap1 is required for Mediator recruitment at these gene promoters and that its recruitment occurs via the tail module. Pol II association is also rapidly and severely affected in srb4 ts yeast, indicating that Mediator is directly required for pol II association at constitutively transcribed genes. Our results are consistent with Mediator functioning as a general transcription factor in yeast.