Miracle-dz/UniFAR-FTUs huggingface dataset:
Comparative analysis of the methods of drug and protein delivery for the treatment of cancer, genetic diseases and diagnostics

The methods of protein and drug delivery for the treatment of cancer, genetic diseases and diagnostics were summarized. The potential of protein transduction is discussed and the recent developments in the field are reviewed. An overview is provided of the non-viral delivery methods such as liposomes, polymer-based delivery, cell-penetrating peptides, bacterial secretion, cells, virosomes, physical methods including electroporation, microinjection, osmotic lysis, nanoparticles, sonoporation to locally inject therapeutic molecules. The characteristic properties of non-viral vectors and their use for the delivery of therapeutic molecules for the diagnosis and treatment of disorders and to target tumors are also discussed. The potential of the transduced peptides and proteins was used as new therapeutic compounds against infectious diseases, to complement deficiencies in specific genes, to specifically kill tumour cells, for gene therapy. The protein delivery vectors can enhance the transfection at low concentrations and help to develop future gene delivery systems with reduced toxicity. Vitamin B12, folic acid, biotin, and riboflavin are essential in the treatment of cancer. Ultrasound has a potential in the delivery of therapeutic agents. The new developing technologies of drug delivery and targeting offer the possibility to improve the therapeutic possibilities of the existing drugs and to develop novel therapeutics.
Regulation of NAD(P)H Oxidase by Associated Protein Disulfide Isomerase in Vascular Smooth Muscle Cells*

NAD(P)H oxidase, the main source of reactive oxygen species in vascular cells, is known to be regulated by redox processes and thiols. However, the nature of thiol-dependent regulation has not been established. Protein disulfide isomerase (PDI) is a dithiol/disulfide oxidoreductase chaperone of the thioredoxin superfamily involved in protein processing and translocation. We postulated that PDI regulates NAD(P)H oxidase activity of rabbit aortic smooth muscle cells (VSMCs). Western blotting confirmed robust PDI expression and shift to membrane fraction after incubation with angiotensin II (AII, 100 nm, 6 h). In VSMC membrane fraction, PDI antagonism with bacitracin, scrambled RNase, or neutralizing antibody led to 26-83% inhibition (p < 0.05) of oxidase activity. AII incubation led to significant increase in oxidase activity, accompanied by a 6-fold increase in PDI refolding isomerase activity. AII-induced NAD(P)H oxidase activation was inhibited by 57-71% with antisense oligonucleotide against PDI (PDIasODN). Dihydroethidium fluorescence showed decreased superoxide generation due to PDIasODN. Confocal microscopy showed co-localization between PDI and the oxidase subunits p22phox, Nox1, and Nox4. Co-immunoprecipitation assays supported spatial association between PDI and oxidase subunits p22phox, Nox1, and Nox4 in VSMCs. Moreover, in HEK293 cells transfected with green fluorescent protein constructs for Nox1, Nox2, and Nox4, each of these subunits co-immunoprecipitated with PDI. Akt phosphorylation, a known downstream pathway of AII-driven oxidase activation, was significantly reduced by PDIasODN. These results suggest that PDI closely associates with NAD(P)H oxidase and acts as a novel redox-sensitive regulatory protein of such enzyme complex, potentially affecting subunit traffic/assembling.
Medication Efficacy: The Impact of Variability in Defining and Measuring ADHD and Executive Functioning

Attention-deficit hyperactivity disorder (ADHD) is a heavily studied topic in neuropsychology and general psychology, psychiatry, and pediatrics. The Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition, Text Revision (DSM-IV) is used to diagnose ADHD, but its criteria are behaviorally defined, while ADHD is a neurological dysfunction. The DSM-IV diagnostic criteria do not differentiate between attention and inhibition or executive functions (EF). As well, there is little agreement in the field about what constitutes ADHD and EFs or how to measure or diagnose them, let alone rule them out. When rendering an ADHD diagnosis, the responsible clinician considers DSM-IV criteria along with other reasons why an individual may express symptoms of ADHD, for reasons other than ADHD. This article discusses postulated mechanisms of action for some medications prescribed for ADHD, the variance of medication outcomes in research, the variance of assessment measures used in identifying ADHD subjects for research, and flaws with DSM-IV–based diagnosis. An argument is posed that the field of neuropsychology needs a cohesive and agreed-upon definition of ADHD and EF and how to measure it to more exactly research the topic, accurately diagnose the disorder, and assist prescribing professionals to use medications with improved first-trial precision.
Debottlenecking recombinant protein production in Bacillus megaterium under large‐scale conditions—targeted precursor feeding designed from metabolomics

In the present work the impact of large production scale was investigated for Bacillus megaterium expressing green fluorescent protein (GFP). Specifically designed scale‐down studies, mimicking the intermittent and continuous nutrient supply of large‐ and small‐scale processes, were carried out for this purpose. The recombinant strain revealed a 40% reduced GFP yield for the large‐scale conditions. In line with extended carbon loss via formation of acetate and carbon dioxide, this indicated obvious limitations in the underlying metabolism of B. megaterium under the large‐scale conditions. Quantitative analysis of intracellular amino acids via validated fast filtration protocols revealed that their level strongly differed between the two scenarios. During cultivation in large‐scale set‐up, the availability of most amino acids, serving as key building blocks of the recombinant protein, was substantially reduced. This was most pronounced for tryptophan, aspartate, histidine, glutamine, and lysine. In contrast alanine was increased, probably related to a bottleneck at the level of pyruvate which also triggered acetate overflow metabolism. The pre‐cursor quantifications could then be exploited to verify the presumed bottlenecks and improve recombinant protein production under large‐scale conditions. Addition of only 5 mM tryptophan, aspartate, histidine, glutamine, and lysine to the feed solution increased the GFP yield by 100%. This rational concept of driving the lab scale productivity of recombinant microorganisms under suboptimal feeding conditions emulating large scale can easily be extended to other processes and production hosts. Biotechnol. Bioeng. 2012; 109:1538–1550. © 2012 Wiley Periodicals, Inc.
Surprising deficiency of CENP-B binding sites in African green monkey alpha-satellite DNA: implications for CENP-B function at centromeres

Centromeres of mammalian chromosomes are rich in repetitive DNAs that are packaged into specialized nucleoprotein structures called heterochromatin. In humans, the major centromeric repetitive DNA, alpha-satellite DNA, has been extensively sequenced and shown to contain binding sites for CENP-B, an 80-kDa centromeric autoantigen. The present report reveals that African green monkey (AGM) cells, which contain extensive alpha-satellite arrays at centromeres, appear to lack the well-characterized CENP-B binding site (the CENP-B box). We show that AGM cells express a functional CENP-B homolog that binds to the CENP-B box and is recognized by several independent anti-CENP-B antibodies. However, three independent assays fail to reveal CENP-B binding sites in AGM DNA. Methods used include a gel mobility shift competition assay using purified AGM alpha-satellite, a novel kinetic electrophoretic mobility shift assay competition protocol using bulk genomic DNA, and bulk sequencing of 76 AGM alpha-satellite monomers. Immunofluorescence studies reveal the presence of significant levels of CENP-B antigen dispersed diffusely throughout the nuclei of interphase cells. These experiments reveal a paradox. CENP-B is highly conserved among mammals, yet its DNA binding site is conserved in human and mouse genomes but not in the AGM genome. One interpretation of these findings is that the role of CENP-B may be in the maintenance and/or organization of centromeric satellite DNA arrays rather than a more direct involvement in centromere structure.
Adipose afferent reflex response to insulin is mediated by melanocortin 4 type receptors in the paraventricular nucleus in insulin resistance rats

Adipose afferent reflex (AAR) contributes to sympathetic activation and hypertension. Paraventricular nucleus (PVN) plays an important role in AAR and sympathetic outflow. The aim of the present study was to determine whether PVN mediates AAR response to insulin in a rat model of insulin resistance (IR).
Pharmacokinetics of repinotan in healthy and brain injured animals

Repinotan hydrochloride (repinotan) is a highly potent and selective 5‐HT1A full receptor agonist. The ability of repinotan to cross the blood–brain barrier (BBB) and penetrate into rat brain tissue was investigated, because rapid penetration into brain tissue is thought to be essential for neuroprotective efficacy. Intravenous (i.v.) repinotan was rapidly distributed into brain, and the distribution equilibrium between blood and brain was reached immediately after the start of infusion. Free concentrations of repinotan were identical in brain and plasma, indicating that repinotan crosses the BBB freely in both directions with diffusion as a driving force. The brain concentration of repinotan was determined by the free plasma concentration. Thus, the total plasma concentration of repinotan (sum of bound and unbound compound) is only indicative for the brain concentration as long as the unbound fraction remains constant. Metabolites of repinotan do not penetrate the BBB and are retained in the perfusing blood due to their increased polarity. The penetration of [14C] repinotan into ischemic areas of the brain was dependent on time. In studies using injured animals (pMCAO), high levels of [14C] repinotan could be detected in ischemic areas when the compound was administered up to 5 h post injury. [14C] repinotan radioactivity could no longer be detected in ischemic areas when administered 18 h after pMCA‐O. After the end of infusion, repinotan was rapidly and completely eliminated from rat brains. Elimination occurred in parallel from plasma and brain with half‐lives of about 1 h. In conclusion, repinotan rapidly and to a considerable extent penetrates into brain tissue of healthy and injured animals. Copyright © 2005 John Wiley & Sons, Ltd.
Low plasma atrial natriuretic peptide: a new piece in the puzzle of polycystic ovary syndrome.

CONTEXT It is believed that a dysfunction in adipose tissue plays an important role in the pathogenesis of polycystic ovary syndrome (PCOS). Natriuretic peptides are hormones that regulate cardiovascular and body fluid homeostasis and adipose tissue metabolism. Natriuretic peptide levels are reduced in individuals with obesity and diabetes. OBJECTIVE This study aimed to investigate whether natriuretic peptide levels are altered in women with PCOS and whether they correlate with adiponectin levels or insulin sensitivity markers. DESIGN AND SETTING This was a cross-sectional study at a referral center in a teaching hospital. PATIENTS OR OTHER PARTICIPANTS We evaluated 40 patients diagnosed with PCOS according to the Rotterdam criteria and 36 control women matched for age and body mass index. MAIN OUTCOME MEASURES We measured serum adiponectin, plasma atrial natriuretic peptide (ANP), and plasma brain natriuretic peptide using enzyme immunoassays in both groups. We evaluated metabolic markers, such as fasting glucose, insulin, total cholesterol, high-density lipoprotein, low-density lipoprotein, and triglycerides. In addition, we calculated the homeostasis model assessment for insulin resistance index (HOMA-IR) and the lipid accumulation product (LAP) index and tested the linear correlations between these metabolic indices and the plasma ANP and serum adiponectin concentrations. RESULTS ANP and adiponectin were reduced in the PCOS group compared with the control group (P = 0.010 and P = 0.014, respectively). The brain natriuretic peptide concentration did not differ between the two groups (P = 0.883). There was no correlation between ANP and any of the metabolic markers. In the control group, the serum adiponectin level was inversely correlated with BMI (P = 0.011), waist circumference (P = 0.021), insulin (P = 0.013), fasting glucose (P = 0.010), homeostasis model assessment for insulin resistance index (P = 0.007), and lipid accumulation product (P = 0.022). Remarkably, none of these correlations were observed in the women with PCOS. CONCLUSION Women with PCOS had lower ANP and adiponectin compared with controls matched for age and BMI. Thus, the mechanisms that affect ANP and adiponectin production and clearance may be altered in PCOS, regardless of adiposity. These hormones may be involved in the metabolic features of PCOS.
Urotensin II is a New Chemotactic Factor for UT Receptor-Expressing Monocytes1

Urotensin II (U-II), a vasoactive cyclic neuropeptide which activates the G protein-coupled receptor UT receptor, exerts various cardiovascular effects and may play a role in the pathophysiology of atherosclerosis. In this study, we report that the UT receptor is expressed and functional on human PBMC and rat splenocytes. PBMC surface expression of the UT receptor was mainly found in monocytes and NK cells, also in a minority of B cells, but not in T cells. Stimulation of monocytes with LPS increased UT receptor mRNA and protein expression. Cloning and functional characterization of the human UT receptor gene promoter revealed the presence of NF-κB-binding sites involved in the stimulation of UT receptor gene expression by LPS. Activation of the UT receptor by U-II induced chemotaxis with maximal activity at 10 and 100 nM. This U-II effect was restricted to monocytes. Analysis of the signaling pathway involved indicated that U-II-mediated chemotaxis was related to RhoA and Rho kinase activation and actin cytoskeleton reorganization. The present results thus identify U-II as a chemoattractant for UT receptor-expressing monocytes and indicate a pivotal role of the RhoA-Rho kinase signaling cascade in the chemotaxis induced by U-II.
Deficiency in APOBEC2 Leads to a Shift in Muscle Fiber Type, Diminished Body Mass, and Myopathy

The apoB RNA-editing enzyme, catalytic polypeptide-like (APOBEC) family of proteins includes APOBEC1, APOBEC3, and activation-induced deaminase, all of which are zinc-dependent cytidine deaminases active on polynucleotides and involved in RNA editing or DNA mutation. In contrast, the biochemical and physiological functions of APOBEC2, a muscle-specific member of the family, are unknown, although it has been speculated, like APOBEC1, to be an RNA-editing enzyme. Here, we show that, although expressed widely in striated muscle (with levels peaking late during myoblast differentiation), APOBEC2 is preferentially associated with slow-twitch muscle, with its abundance being considerably greater in soleus compared with gastrocnemius muscle and, within soleus muscle, in slow as opposed to fast muscle fibers. Its abundance also decreases following muscle denervation. We further show that APOBEC2-deficient mice harbor a markedly increased ratio of slow to fast fibers in soleus muscle and exhibit an ∼15–20% reduction in body mass from birth onwards, with elderly mutant animals revealing clear histological evidence of a mild myopathy. Thus, APOBEC2 is essential for normal muscle development and maintenance of fiber-type ratios; although its molecular function remains to be identified, biochemical analyses do not especially argue for any role in RNA editing.
Misrepresentation of horizontal space in left unilateral neglect

Background: Right-brain–damaged patients with left unilateral neglect are reported to misperceive the horizontal extension of contralesional stimuli as being shorter than that of ipsilesional stimuli. Objective: To investigate the functional and anatomic correlates of horizontal space misrepresentation. Methods: Eight right-brain–damaged patients with contralesional neglect and complete hemianopia (N+H+), nine right-brain–damaged patients with contralesional neglect and no visual field defect (N+H−), and five unilateral brain-damaged patients with contralesional complete hemianopia and no neglect (N−H+) reproduced a horizontal distance (10 cm) in the contralesional and ipsilesional hemispace. Results: N+H+ patients overextended the distance contralesionally and underextended the same distance ipsilesionally. N+H− and N−H+ patients reproduced equivalent distances contralesionally and ipsilesionally. Compared with N+H− patients, N+H+ patients had a greater ipsilesional shift when bisecting horizontal lines; however, these two groups of patients had comparable neglect severity on multiple-item cancellation tasks. In the N+H+ group the area of maximal overlapping of the lesion was in the posterior cerebral lobes. Conclusion: Complete contralesional hemianopia after posterior brain damage is an important factor in determining misrepresentation of horizontal space in patients with left unilateral neglect.
Rescue of photoreceptors from the damaging effects of constant light by midkine, a retinoic acid-responsive gene product.

PURPOSE To evaluate the protective effects of midkine (MK), the product of a retinoic acid-responsive gene, on constant light-induced retinal degeneration in albino Sprague-Dawley rats. METHODS Midkine, basic fibroblast growth factor (bFGF), MK plus heparin, or buffer controls were injected intravitreally 2 days before constant light exposure. After 7 days of continuous light exposure, the eyes were perfused with fixative, bisected along the vertical meridian, embedded in paraffin, and sectioned. The degree of retinal light damage was assessed for paraffin-embedded sections by cytologic analysis, by measuring the thickness of the outer nuclear layer (ONL), and by counting the number of macrophages. RESULTS After 1 week of constant light exposure, uninjected controls and those injected with phosphate-buffered saline (PBS) lost most of the photoreceptor inner and outer segments, and the thickness of the ONL was decreased. Eyes that were injected with MK or bFGF demonstrated a significant rescue in the photoreceptor layer with a two- to threefold increase in the ONL thickness. The number of macrophages in eyes injected with MK was significantly suppressed compared with controls. Those injected with bFGF had a 1.5-fold increase in number compared with controls. CONCLUSIONS Midkine has shown strong survival-promoting activity in constant light-induced retinal degeneration, and thus has a high degree of neurotrophic activity in vivo.
[Screening and identification of heterogeneous phenotype-associated genes in human bladder cancer].

OBJECTIVE To screen and identify heterogeneous phenotype-associated genes of human bladder transitional cell carcinoma. METHODS Subtractive cDNA libraries was established by means of suppression subtractive hybridization (SSH) on the basis of two human bladder transitional cell carcinoma cell lines (BLZ-211 and BLS-211) derived from the same patient, which had similar changes in chromosomes but different cell phenotypes (in terms of cell shape, susceptibility to 5-Fu and tumorigenic capacity). The positive clones in the library were selected for screening differentially expressed gene fragments by sequence analysis and blasting, and Northern blotting was performed to confirm the differentially expressed genes. RESULTS The subtractive forward and reverse cDNA libraries consisted of 168 and 206 white clones containing 200-700 bp inserts. After differential screening, 55 cDNA clones containing 35 different transcripts were obtained, among which 15 were identified by homology analysis as known genes (such as those coding for vimentin, keratin 7, dihydrodiol dehydrogenase and thioredoxin reductase), 11 as unknown genes, and 9 as new ESTs (GenBank dbEST database accession number DY505708, DY230447-8, DR008207). CONCLUSION SSH is a powerful method for identifying differentially expressed genes between different cell lines or clones, and characterization of the identified genes may provide useful information for understanding the genes responsible for different cell phenotypes.
Analysis of the P Wave in the Signal‐Averaged Electrocardiogram: Normal Values and Reproducibility

To apply p wave analysis of the signal‐averaged electrocardiogram (SAECG) clinically, the knowledge of normal values and the verification of highly reproducible measurements are essential. In 40 healthy volunteers (ages 23 ‐37 years), an SAECG of the p wave was performed and then repeated after 1 week and 1 month. In addition to Simson analysis (unidirectional and bidirectional filters), the data were filtered with finite impulse response (FIR) and least squares fit (LSQ) filters to obtain a better differentiation between the end of the p wave and QRS onset. All recordings were evaluated by two independent observers, and the following parameters were calculated: the duration of the p wave in the three unfiltered leads; and for the vector magnitude of the three leads x, y, and z: the duration of the p wave and the root mean square voltage of its last 30, 20, and 10 ms. The average p wave duration was significantly different among the various filter techniques: 135 ± 7 ms (unidirectional); 97 ± 8 ms (bidirectional); 109 ± 8.5 ms (FIR); and 126 ± 10 ms (LSQ) (P = 0.001 for each comparison). There was a good reproducibility of the data analyzed with the Simson method (R = 0.80–0.91). Filtering with the FIR and LSQ leads to a considerably greater variability.
Estimating incidence and prevalence of episodes of care in general practice

Objectives To develop methods for prospective registration and analysis of episodes of care (one or more contacts about the same health problem). To compare estimates of point prevalence and incidence of hypertension among a group of elderly patients by means of an elaborate coding of episodes and a new method based on routine coding of contact diagnoses. Design Prospective longitudinal cohort study. Setting General practice in Denmark. Subjects 31 GPs and 1722 patients 70+ years of age. Main outcome measures During the course one year, data were registered in the doctors’ electronic medical record systems. Registration included ICPC diagnoses and codes for diagnostic and therapeutic procedures. Individual contacts were linked into episodes of care by the recording physician. Prevalence and incidence were calculated from the GPs’ registration of episodes of care and from contact registration by means of a new method, “the waiting time distribution”. Results Estimates of incidence (2.1 per 100 patient years) and point prevalence (21.2 per 100 patients) of episodes of hypertension care could be calculated when the GPs actively linked contacts into episodes. Based on simple contact registration it was possible to calculate similar estimates. Conclusion A full registration of episodes of care is time-consuming and complicated. Incidence and point prevalence of chronic conditions, however, may be estimated from simple coding of contact diagnoses.