Miracle-dz/UniFAR-FTUs huggingface dataset:
Expression of metallothionein isoforms in human chorioretinal complex

Purpose. To determine the relative expression of metallothionein isoforms and their differential induction by oxidative stress in cultured RPE cells and to localize the isoforms in the human chorioretinal complex. Methods. Total RNA was isolated from cultured human retinal pigment epithelial cells using TRI-Reagent. An “anchor-oligo-dT primer” was used for the synthesis of cDNA, reverse transcribed using avian reverse transcriptase and subsequently subjected to PCR analysis using oligonucleotides specific for metallothionein (MT) I, MT II, and MT III. The selected transcripts were then used to assess the expression of the above elements in fixed tissue sections by in situ hybridization. Cultured RPE cells were allowed to phagocytose bovine photoreceptor outer segments (ROS) or were treated with H 2 O 2 for 6 hours and then analyzed by RT-PCR or in situ hybridization to ascertain the effect of oxidative stress on metallothionein mRNA isoform expression. Results. Relative density analysis of amplified products demonstrate the presence of MT I, MT II and MT III in RPE cells, with an apparent relative expression MT II < MT I < MT III. Expression of MT I and MT II mRNA was increased by both phagocytosis and hydrogen peroxide, however MT III was not induced by either stress. In situ hybridization corroborated the findings of the RT-PCR analysis and showed that MTs were mainly localized in the RPE and the photoreceptor layer of the retina. Conclusions. The localization of MT and the response of MT to oxidative stress are consistent with a role for MT as an antioxidant in the RPE and retina. Studies are ongoing to determine the specific mechanisms of action of these antioxidants in RPE cells.
The Scc Spirochetal Coiled-Coil Protein Forms Helix-Like Filaments and Binds to Nucleic Acids Generating Nucleoprotein Structures

ABSTRACT The analysis of the genome of Leptospira spp. , a group of bacteria of the phylum of spirochetes with several unique evolutionary and morphological features, has allowed the identification of a gene encoding a coiled-coil protein, called Scc, which is completely unrelated to any other eukaryotic or prokaryotic protein. Since coiled-coil proteins are often key elements of the cytoskeleton, we analyzed the protein Scc, which is a 24-kDa protein composed of a N-terminal coiled-coil domain, a proline-rich intermediate domain, and an acidic tail. The gene scc is located in an operon which also contains the genes encoding the initiation factor IF3 and the two ribosomal proteins L20 and L35. In this study, we showed that the presence of the coiled-coil domain was responsible for the polymerization of Scc in helix-like structures, in an ATP-independent manner, in both Escherichia coli living cells and in vitro. Analysis of the Scc polymers by electron microscopy showed filaments with a width of 6 to 10 nm, similar to that of eukaryotic intermediate filaments. Scc was also found to bind both RNA and double-stranded DNA without detectable sequence specificity. By electron microscopy, we showed that Scc polymer assembly was affected by the presence of nucleic acids, giving rise to rod-shaped structures with a width ranging from 45 to 155 nm. Finally, Leptospira biflexa cells depleted in Scc form small colonies, but the morphology of their helicoidal cell body was not affected. These results provide the first insight into a unique DNA binding filament-forming coiled-coil protein that could play an important role in the subcellular architecture of the spirochetal microorganism.
Statistical analysis of multi‐subject fMRI data: Assessment of focal activations

A simple procedure for analyzing multi‐subject functional magnetic resonance imaging (fMRI) data is proposed. In the first step, a voxel‐wise t‐test across standardized z‐maps is performed to identify areas that are consistently activated across subjects. In the second step, for each area, individual mean z‐scores are calculated and subsequently subjected to an analysis of variance. An example is provided. J. Magn. Reson. Imaging 2000;11:61–64. © 2000 Wiley‐Liss, Inc.
Limited Sampling Strategies for Therapeutic Drug Monitoring of Linezolid in Patients With Multidrug-Resistant Tuberculosis

Introduction: Linezolid is a potential drug for the treatment of multidrug-resistant tuberculosis but its use is limited because of severe adverse effects such as anemia, thrombocytopenia, and peripheral neuropathy. This study aimed to develop a model for the prediction of linezolid area under the plasma concentration-time curve from 0 to 12 hours (AUC0-12h) by limited sampling strategy to enable individualized dosing. Patients and Methods: Fourteen patients with multidrug-resistant tuberculosis received linezolid twice daily as part of their antituberculosis treatment. Linezolid concentrations were determined at steady state by high-performance liquid chromatography tandem mass spectrometry before and at 1, 2, 4, 8, and 12 hours after dosing. Linezolid AUC0-12h population model and limited sampling models were calculated with MWPharm software. The correlation between predicted linezolid AUC0-12h and observed linezolid AUC0-12h was investigated by Bland-Altman analysis. Results: A total of 26 pharmacokinetic profiles were obtained. The median AUC0-12h was 51.8 (interquartile range, 41.8-65.9) mg*h/L at 300 mg and 123.8 (interquartile range, 100.9-152.5) mg*h/L at 600 mg, both twice daily. The most relevant model clinically for prediction of linezolid AUC0-12h used a linezolid trough concentration (r = 0.91, prediction bias = -2.9% and root mean square error = 15%). Discussion: The difference between choosing a trough concentration and two to three samples increased the correlation from 0.90 to 0.95 but appeared not clinically relevant because it did not result in different dosing advice. Conclusion: This study showed that linezolid AUC0-12h in patients with multidrug-resistant tuberculosis could be predicted accurately by a minimal sampling strategy and could be used to individualize the dose.
Ligand‐induced changes in the conformational stability and flexibility of glutamate dehydrogenase and their role in catalysis and regulation

Bovine glutamate dehydrogenase (GDH) is allosterically regulated and requires substrate‐induced subunit interactions for maximum catalytic activity. Steady‐state and presteady‐state kinetics indicate that the rate‐limiting step depends on the nature of the substrate and are likely associated with conformational fluctuations necessary for optimal hydride transfer. Deuterated glutamate shows a steady‐state isotope effect but no effect on the presteady‐state burst rate, demonstrating that conformational effects are rate limiting for hydride transfer while product release is overall rate limiting for glutamate. Guanidine hydrochloride unfolding, heat inactivation, and differential scanning calorimetry demonstrate the effects of alternative substrates, glutamate and norvaline, on conformational stability. Glutamate has little effect on overall stability, whereas norvaline markedly stabilizes the protein. Limited proteolysis demonstrates that glutamate had a variety of effects on local flexibility, whereas norvaline significantly decreased conformational fluctuations that allow protease cleavage. Dynamic light scattering suggests that norvaline stabilizes all interfaces in the hexamer, whereas glutamate had little effect on trimer–trimer interactions. The substrate glutamate exhibits negative cooperativity and complex allosteric regulation but has only minor effects on global GDH stability, while promoting certain local conformational fluctuations. In contrast, the substrate norvaline does not show negative cooperativity or allow allosteric regulation. Instead, norvaline significantly stabilizes the enzyme and markedly slows or prevents local conformational fluctuations that are likely to be important for cooperative effects and to determine the overall rate of hydride transfer. This suggests that homotropic allosteric regulation by the enzymatic substrate involves changes in both global stability and local flexibility of the protein.
A Sociological Approach to Police Officers’ Conflict Management Tools

Conflicts are inseparable part of human society all over the world. Although much contemporary conflicts seem to be about political, ethnic, or religious differences, in fact these conflicts generally have an economic, and a political basis. One of the most important nowadays issue is how people manage conflict situations, driven by interests, seemingly incompatible values and hostilities. Albanian State Police strategies vision is supporting new arrangements and innovation about police communication services. In addition, it appears necessary for police officers to be prepared for effective intervention and conflict resolution. The objective of this research is to examine conflict situations and relationships in police institution. The aim is to find out which appropriate methods of analysis or management strategies are better for solutions between parties of conflicts. The method used for the conduction of this work is a qualitative one, combined with analyses of data of a short survey process. Through the literature research and analyses of the data collected from the survey there are provided ways or manners of dealing and managing conflicts situations between police officers, people and police officers or police officers in front of people who conflict to each other. Analysis of the collected data have revealed in the conclusion that in order to perfectly resolve and manage conflict situations, it’s necessary to primary deeply analyze situations. This works in police officers case, too. As a main conclusion of this paper, each police officer who deals with a conflict situation has to structure their work plan and has to mention the main points like: description of the situation, identification of six phases of a conflict situation, deepening analysis based on the “mapping” method, following police procedures and finding a path for an adequate management and solution. Mapping method is used in terms of in-depth analysis of conflict situations to bring out better understanding of conflicts causes and parties.
DNA damage‐dependent NF‐κB activation: NEMO turns nuclear signaling inside out

Summary: The dimeric transcription factor nuclear factor κB (NF‐κB) functions broadly in coordinating cellular responses during inflammation and immune reactions, and its importance in the pathogenesis of cancer is increasingly recognized. Many of the signal transduction pathways that trigger activation of cytoplasmic NF‐κB in response to a broad array of immune and inflammatory stimuli have been elaborated in great detail. NF‐κB can also be activated by DNA damage, though relatively less is known about the signal transduction mechanisms that link DNA damage in the nucleus with activation of NF‐κB in the cytoplasm. Here, we focus on the conserved signaling pathway that has emerged that promotes NF‐κB activation following DNA damage. Post‐translational modification of NF‐κB essential modulator (NEMO) plays a central role in linking the cellular DNA damage response to NF‐κB via the ataxia telangiectasia mutated (ATM) kinase. Accumulating evidence suggests that DNA damage‐dependent NF‐κB activation may play significant biological roles, particularly during lymphocyte differentiation and progression of human malignancies.
Post‐training intrahippocampal infusion of nicotine prevents spatial memory retention deficits induced by the cyclo‐oxygenase‐2‐specific inhibitor celecoxib in rats

Recently, we demonstrated that intrahippocampal infusion of the cyclo‐oxygenase (COX)‐2‐specific inhibitor celecoxib impaired spatial memory retention in the Morris water maze. In the present work, we investigated the effects of nicotine, infused in the rat dorsal hippocampus several minutes after infusion of celecoxib, on memory retention in the Morris water maze. Rats were trained for 3 days; each day included two blocks, and each block contained four trials. Test trials were conducted 48 h after surgery. As expected, bilateral intrahippocampal infusion of celecoxib (19 µg/side; 0.1 m) increased escape latency and travel distance in rats, indicating significant impairment of spatial memory retention. We also examined the effects of bilateral infusion of nicotine (0.5, 1.0 and 2.0 µg/side) on memory retention. Infusion of 1 µg nicotine significantly decreased escape latency and travel distance but not swimming speed, compared with controls, suggesting memory retention enhancement by nicotine at this concentration. In separate experiments, bilateral infusion of nicotine, infused 5 min after 0.1 m (19 µg/side) celecoxib infusion, was associated with escape latency, travel distance and swimming speed profiles very similar to those in control animals. Brain tissue sections from several of these animals were subjected to immunohistochemical staining analysis with anti‐COX‐2 antibodies. Quantification analysis by optical density measurements showed that the celecoxib infusion reduced the immunoreactivity of COX‐2‐containing neurons in the CA1 area of the hippocampus compared with controls, although this reduction was not significant. However, infusion of a combination of celecoxib and nicotine significantly increased this immunoreactivity compared with levels in control and celecoxib‐infused groups. These results suggest that nicotine prevented or reversed the adverse effects of celecoxib on spatial memory retention and protected or restored the immunostaining pattern of COX‐2 neurons in the rat dorsal hippocampus.
Social media driven innovations: an exploratory study in China

As societies move into cyberspace, the exponential growth of Social Media (SM) is enabling business innovations. Yet little knowledge exists on the role SM plays in driving innovations in Chinese businesses. Addressing this gap, data collected through semi-structured interviews were analyzed using content analysis techniques. We find that SM is driving innovations in: (1) agile product testing/trials to eliminate or reduce re-work/waste, implicitly facilitating lean management, (2) development of payment and distribution channels including SM enabled Online-to-Offline business, (3) empowered integration of ‘wisdom of masses’ in product/service design and delivery processes, (4) farming seeds of consumer circles for marketing/sales and enhanced loyalty, (5) development of social media shop fronts, and (6) development of agile product design teams transcending organizational talent boundaries. Academically, new findings extend theories on innovation, particularly open innovations. Managerially, the findings provide strategies for business/IT executives to leverage SM to drive innovations, and policy makers in developing policies that facilitate SM-driven innovations.JEL Codes: O14, L86, N75
Neuronal expression of tachykinin‐related peptides and gene transcript during postembryonic development of Drosophila

The gene Dtk, encoding the prohormone of tachykinin‐related peptides (TRPs), has been identified from Drosophila. This gene encodes five putative tachykinin‐related peptides (DTK‐1 to 5) that share the C‐terminal sequence FXGXRamide (where X represents variable residues) as well as an extended peptide (DTK‐6) with the C‐terminus FVAVRamide). By mass spectrometry (MALDI‐TOF‐MS), we identified ion signals with masses identical to those of DTK‐1 to 5 in specific brain regions. We have analyzed the distribution of the Dtk transcript and peptides, by in situ hybridization and immunocytochemistry during postembryonic development of the central nervous system (CNS) of Drosophila. Antiserum against a cockroach TRP that cross‐reacts with the DTKs was used for immunocytochemistry. Expression of transcript and peptides was detected from first to third instar larvae, through metamorphosis to adult flies. Throughout postembryonic development, we were able to follow the strong expression of TRPs in a pair of large descending neurons with cell bodies in the brain. The number of TRP‐expressing neuronal cell bodies in the brain and ventral nerve cord increases during larval development. In the early pupa (stage P8), the number of TRP‐expressing cell bodies is lower than in the third instar larvae. The number drastically increases during later pupal development, and in the adult fly about 200 TRP‐expressing neurons can be seen in the CNS. The continuous expression of TRPs in neurons throughout postembryonic development suggests specific functional roles in both larval and imaginal flies and possibly also in some neurons during pupal development. J. Comp. Neurol. 464:180–196, 2003. © 2003 Wiley‐Liss, Inc.
The use of framing of affect or reason in communications in a services failure recovery scenario in education

The study of decision-making of people has its origins in economic theory, with a more rational approach. However, several studies have shown that decision-making follows also an emotional model. In great part, decisions are influenced by information we receive through communication framing. In education, students decisions are largely affected by the information they receive through communication issued by the service provider. In a scenario of service failure recovery the influence of emotional or rational messages is little studied. The motivation of this study is the absence of works relating attitudes to services when failure recovery occurs in higher education services, particularly when the framing of the communication signals some position to students, aiming to persuade them. The results showed that rational communication was more effective than emotional ones. It was also found that interpersonal influences tends to reduce positive responses from students to HEI communication strategy, though a moderation process.
Improving coverage and connectivity using harmony search algorithm in wireless sensor network

The aim of this paper has been to improve WSN's coverage and connectivity besides reduction in the number of sensor nodes deployed in the ROI (Region of Interest). Here, the coverage deals with maximizing the area covered by the sensing range of sensor in the ROI and the connectivity means that each and every sensor node should be under the communication range of at least one sensor node. To achieve this objective a Harmony Search algorithm has been utilized and simulation has been performed to obtain improved coverage and connectivity. Average Coverage ratio, Average Connectivity ratio and Average Sensing ratio has been considered as the performance metrics. It is observed that average coverage ratio and average connectivity ratio is maximum when communication range is double the sensing range. The average coverage ratio has been improved by 16.95% using proposed technique in comparison to the existing Harmony Search approach.
One-Year Experience with Day-of-Surgery Pregnancy Testing Before Elective Orthopedic Procedures

BACKGROUND: Elective surgery is generally postponed in pregnancy. A policy of testing for urine human chorionic gonadotropin (hCG) in all women of childbearing age on the day of surgery was initiated at an elective orthopedic surgery facility. This is a retrospective report of our 1 yr experience and the associated costs. METHODS: Records were reviewed from January 3, 2005, through January 2, 2006, to determine the number of urine hCG tests performed, and the disposition of all patients with a positive result. Costs were calculated using the charges for testing supplies and laboratory labor. RESULTS: During the first year of policy implementation, 2588 of 2595 women of childbearing age underwent urine hCG testing. Five patients had a positive result, and surgery was cancelled. Of these five, three were previously unrecognized pregnancies, one an unrecognized asymptomatic ectopic pregnancy, and one a false-positive result in a perimenopausal woman. Three other women had a “weak positive” urine result, followed by a negative serum hCG. Surgery proceeded in all three. The calculated cost was $5.03 per urine test, and $3273 for each true positive result. CONCLUSIONS: A policy of routinely performing urine hCG pregnancy tests in women of childbearing age on the day of surgery was effective in detecting unrecognized pregnancy. This resulted in a postponement of elective surgery in all cases. Of 2588 women tested, one had surgery postponed because of a false-positive result. The cost of $3273 per true positive test must be compared with the benefit.
IGF‐1 increases laminin, cyclin D1, and P21Cip1 expression in glomerular mesangial cells: An investigation of the intracellular signaling pathway and cell‐cycle progression

Insulin‐like growth factor (IGF)‐1 is accumulated in the diabetic kidney and is considered to be involved in the development of glomerular sclerosis. Here, we investigate IGF‐1 regulation of laminin, an extracellular matrix (ECM) component, and cyclin D1 and p21Cip1, cell‐cycle progression factor, expressions in glomerular mesangial cells. We show that IGF‐1 increases the level of laminin γ1 and β1 subunits ∼1.5‐ and 2.5‐fold, respectively, in a time‐dependent manner. IGF‐1 also stimulates protein kinase Akt/PKB phosphorylation at Thr 308, which correlates with its activity, up to 24 h. The Akt activation is coupled with Ser 9 phosphorylation of its downstream target, glycogen synthase kinase‐3beta (GSK‐3β), which inhibits its kinase activity. Laminin β1 is reduced significantly (P < 0.03) by inhibitors of Akt and p38MAPK whereas laminin γ1 is not affected. Surprisingly, IGF‐1 activates the expression of both cyclin D1 and cell‐cycle arrest factor, p21Cip1 parallely. Pharmacological inhibition of calcineurin by cyclosporin A blocks IGF‐1‐induced cyclin D1 and p21Cip1expression significantly (P < 0.05). IGF‐1 enhances cellular metabolic activity and viability of rat mesangial cells; however, they are arrested at the G1 phase of cell cycle as revealed by the FACS analysis. These results indicate that IGF‐1 mediates mesangial cell‐cycle progression, hypertrophy, and ECM protein synthesis. The Akt/GSK‐3β, p38MAPK, and calcineurin pathways may play an important role in IGF‐1 signaling, cell‐cycle regulation, and matrix gene expression in mesangial cells leading to the development of diabetic glomerulopathy. J. Cell. Biochem. 98: 208–220, 2006. © 2006 Wiley‐Liss, Inc.
A mutation within the C‐terminal domain of Sup35p that affects [PSI+] prion propagation

The epigenetic factor [PSI+] in the yeast Saccharomyces cerevisiae is due to the prion form of Sup35p. The N‐terminal domain of Sup35p (N), alone or together with the middle‐domain (NM), assembles in vitro into fibrils that induce [PSI+] when introduced into yeast cells. The Sup35p C‐terminal domain (C), involved in translation termination, is essential for growth. The involvement of Sup35p C‐terminal domain into [PSI+] propagation is subject to debate. We previously showed that mutation of threonine 341 within Sup35p C‐domain affects translation termination efficiency. Here, we demonstrate that mutating threonine 341 to aspartate or alanine results in synthetic lethality with [PSI+] and weakening of [PSI+] respectively. The corresponding Sup35D and Sup35A proteins assemble into wild‐type like fibrils in vitro, but with a slower elongation rate. Moreover, cross‐seeding between Sup35p and Sup35A is inefficient both in vivo and in vitro, suggesting that the point mutation alters the structural properties of Sup35p within the fibrils. Thus, Sup35p C‐terminal domain modulates [PSI+] prion propagation, possibly through a functional interaction with the N and/or M domains of the protein. Our results clearly demonstrate that Sup35p C‐terminal domain plays a critical role in prion propagation and provide new insights into the mechanism of prion conversion.