Dimethyl Fumarate Reduces Microglia Functional Response to Tissue Damage and Favors Brain Iron Homeostasis.
Dimethyl fumarate (DMF) is the only available approved drug for first line treatment of multiple sclerosis (MS), a lethal condition impairing central nervous system (CNS). To date, however, little is known of its mechanisms of action. Only recently, it has been suggested that DMF exerts neuroprotective effects acting as an immunomodulator and that it may alter the activation state of microglia cells, crucial in MS pathogenesis. However, DMF effects on microglia functions are still not well determined. Here, we examine the effects of DMF treatment on microglia functional activities, as phenotype, morphology, processes motility and rearrangement, migration, ATP response and iron uptake in mouse primary microglia culture and acute hippocampal slices. We found that DMF treatment reduces microglia motility, downregulating functional response to ATP, increases ferritin uptake and pushes microglia towards an anti-inflammatory phenotype, thus reducing its proinflammatory reactivity in response to tissue damage. These results highlight the effects of this compound on microglia functions and provide new insights on the mechanism of action of DMF in MS treatment.


Enterococcus faecium are associated with the modification of gut microbiota and shrimp post-larvae survival.
Probiotics are widely used to promote host health. Compared to mammals and terrestrial invertebrates, little is known the role of probiotics in aquatic invertebrates. In this study, eighteen tanks with eight hundred of shrimp post-larvae individuals each were randomly grouped into three groups, one is shrimps administered with E. faecium as probiotic (Tre) and others are shrimps without probiotic-treatment (CK1: blank control, CK2: medium control). We investigated the correlations between a kind of commercial Enterococcus faecium (E. faecium) powder and microbiota composition with function potentials in shrimp post-larvae gut. We sequenced the 16S rRNA gene (V4) of gut samples to assess diversity and composition of the shrimp gut microbiome and used differential abundance and Tax4Fun2 analyses to identify the differences of taxonomy and predicted function between different treatment groups. The ingested probiotic bacteria (E. faecium) were tracked in gut microbiota of Tre and the shrimps here showed the best growth performance especially in survival ratio (SR). The distribution of SR across samples was similar to that in PCoA plot based on Bray-Curits and two subgroups generated (SL: SR < 70%, SH: SR ≥ 70%). The gut microbiota structure and predicted function were correlated with both treatment and SR, and SR was a far more important factor driving taxonomic and functional differences than treatment. Both Tre and SH showed a low and uneven community species and shorted phylogenetic distance. We detected a shift in composition profile at phylum and genus level and further identified ten OTUs as relevant taxa that both closely associated with treatment and SR. The partial least squares path model further supported the important role of relevant taxa related to shrimp survival ratio. Overall, we found gut microbiota correlated to both shrimp survival and ingested probiotic bacteria (E. faecium). These correlations should not be dismissed without merit and will uncover a promising strategy for developing novel probiotics through certain consortium of gut microbiota.


Validation of Stroke Meaningful Use Measures in a National Electronic Health Record System.
The Meaningful Use (MU) program has increased the national emphasis on electronic measurement of hospital quality. To evaluate stroke MU and one VHA stroke electronic clinical quality measure (eCQM) in national VHA data and determine sources of error in using centralized electronic health record (EHR) data. Our study is a retrospective cross-sectional study of stroke quality measure eCQMs vs. chart review in a national EHR. We developed local SQL algorithms to generate the eCQMs, then modified them to run on VHA Central Data Warehouse (CDW) data. eCQM results were generated from CDW data in 2130 ischemic stroke admissions in 11 VHA hospitals. Local and CDW results were compared to chart review. We calculated the raw proportion of matching cases, sensitivity/specificity, and positive/negative predictive values (PPV/NPV) for the numerators and denominators of each eCQM. To assess overall agreement for each eCQM, we calculated a weighted kappa and prevalence-adjusted bias-adjusted kappa statistic for a three-level outcome: ineligible, eligible-passed, or eligible-failed. In five eCQMs, the proportion of matched cases between CDW and chart ranged from 95.4 %-99.7 % (denominators) and 87.7 %-97.9 % (numerators). PPVs tended to be higher (range 96.8 %-100 % in CDW) with NPVs less stable and lower. Prevalence-adjusted bias-adjusted kappas for overall agreement ranged from 0.73-0.95. Common errors included difficulty in identifying: (1) mechanical VTE prophylaxis devices, (2) hospice and other specific discharge disposition, and (3) contraindications to receiving care processes. Stroke MU indicators can be relatively accurately generated from existing EHR systems (nearly 90 % match to chart review), but accuracy decreases slightly in central compared to local data sources. To improve stroke MU measure accuracy, EHRs should include standardized data elements for devices, discharge disposition (including hospice and comfort care status), and recording contraindications.


Heterogeneity in subcutaneous adipose tissue morphology and metabolic complications in overweight and obese women.
The aim of this study was to assess morphological features of intact adipose tissue (AT) ex vivo from both subcutaneous (s.c.) abdominal and gluteal areas using a novel approach of multiphoton autofluorescence microscopy (MPAM) combined with second harmonic generation microscopy (SHGM), and to assess the relationship between morphological features in the two AT sites and insulin resistance to peripheral glucose disposal. This study was a cross-sectional evaluation of AT morphology feature and peripheral insulin resistance. Fourteen overweight/obese premenopausal women underwent body composition studies, hyperinsulinemic-euglycemic clamps, and needle biopsy of both the s.c. abdominal and gluteal AT areas. MPAM combined with SHGM was used to measure adipocyte maximal diameter and collagen fiber bundle thickness within a sampled image volume after three-dimensional visualization. Higher body mass index (BMI) was associated with larger adipocyte diameter in s.c. abdominal, but not gluteal, AT. Higher adipocyte diameter was associated with higher pericellular collagen thickness. Adipocyte diameter in s.c. abdominal, but not gluteal, AT was associated positively with leptin and negatively with adiponectin plasma levels and peripheral glucose disposal rate. The latter correlation was no longer significant after adjustment for collagen thickness. In overweight/obese premenopausal women, larger adipocyte diameter in s.c. abdominal, but not gluteal, AT associates with low plasma adiponectin and systemic insulin resistance, and suggests that increased collagen thickness (obesity-related scarring) could contribute to these findings.


Determination of digoxin clearance in Japanese elderly patients for optimization of drug therapy: a population pharmacokinetics analysis using nonlinear mixed-effects modelling.
Optimal use of digoxin in the elderly population requires information about the drug's pharmacokinetics and the influence of various factors on the drug's disposition. However, because of sampling restrictions, it is often difficult to perform traditional pharmacokinetic studies in elderly patients. This study was conducted to determine the apparent total clearance of digoxin from serum after oral administration (CL/F) and to establish the role of patient characteristics in estimating doses of digoxin for elderly patients (age ≥65 years), using routine therapeutic drug monitoring data. Analyses of the pharmacokinetics of digoxin were conducted using the nonlinear mixed-effects modelling (NONMEM®) software, a computer program designed to analyse pharmacokinetics in study populations by allowing pooling of data. Steady-state data (140 observations) obtained by routine therapeutic drug monitoring following repeated oral administration of digoxin in 94 hospitalized elderly patients (age ≥65 years) were analysed to establish the role of patient characteristics in estimating doses of digoxin for elderly patients. Estimates generated by NONMEM® indicated that digoxin CL/F was influenced by the demographic variables of total bodyweight (TBW), serum creatinine (SCr), age (AGE), presence of congestive heart failure (CHF), concomitant administration of the calcium channel antagonists (calcium channel blockers [CCBs]: verapamil, diltiazem or nifedipine), sex (SEX) and elderly clearance factor (trough serum concentration of digoxin; [C(trough)] θ). The full version of the final NONMEM® model was where CCB is 1 for concomitant administration of a CCB and is 0 otherwise; CHF is 1 for patients with CHF and is 0 otherwise; SEX is 0 for male and is 1 for female; and the elderly clearance factor C(trough)-0.180 is 1 for digoxin C(trough) <1.7 ng/mL. We developed a new model for elderly patient dosing of digoxin with good predictive performance. Clinical application of the findings of the present study to patient care may permit selection of an appropriate initial digoxin maintenance dose, thus enabling the clinician to achieve a desired therapeutic effect. However, the digoxin dosage regimen should be based on an appraisal of the individual patient's clinical need for the drug.


In Vitro Evaluation of Aortic Stent Graft Deployment Accuracy in the Distal Landing Zone.
Thoracic endovascular aortic repair technology focuses on accurately deploying the stent graft in the proximal landing zone in the proximal to distal direction. The aim of this in vitro study was to evaluate the accuracy of stent graft deployment in the distal landing zone. It was hypothesised that a reverse implantation mechanism (in a distal to proximal deployment direction, when the operator starts to open the endoprosthesis from distal to proximal), might enhance landing accuracy in the distal LZ. The aim was to investigate this hypothesis by implanting stent grafts into the 3D printed aortas with the currently available deployment mechanism. Based on two human patients' computed tomography angiography scans, two aortas were 3D printed at 1:1 scale: "straight" and "crooked" aortas with distal aortic tortuosity of 1.006 and 1.078, respectively. They were used in order to test three endoprostheses (E-vita THORACIC 3G, Relay Plus, Valiant Captivia) 10 times by implanting them in three ways: proximal landing in the aneurysm, proximal landing in another stent graft, reverse implantation (via simulated antegrade access). The aim was to land just above the target vessel's upper edge. The distance to the target vessel and wedge apposition were assessed under a direct view using caliper. The distance to the target vessel was 3 mm (IQR 0; 8) if the stent graft landed proximally in aneurysm, 2 mm (IQR 0; 5) if it landed proximally in another stent graft, and 0 mm (IQR 0; 0) when reverse implantation was applied. The distance to the target vessel measuring 5 mm or occurred in 45%, 30%, and 0%, respectively. Overall the median wedge apposition after stent graft implantation was 0 mm (IQR 0; 0) in the "straight" versus 18 mm (IQR 15; 20) in the "crooked" aorta (p < .001). Reverse stent graft deployment is associated with more accurate landing in the distal landing zone. Distal aortic tortuosity constitutes an important impediment to covering the distal LZ's entire circumference with a stent graft.


Corrigendum to "Effect of vermicompost on vegetative growth and nutrient status of acclimatized Grand Naine banana plants" [Heliyon 8 (10), October 2022, Article e10914].
[This corrects the article DOI: 10.1016/j.heliyon.2022.e10914.].


Erythrocyte aldose reductase protein: a clue to elucidate risk factors for diabetic neuropathies independent of glycemic control.
Prolonged hyperglycemia has been thought to be the primary cause of diabetic complications, however, some diabetic patients develop severe complications early in duration of diabetes, while some patients have no or mild complications even after prolonged hyperglycemia. To investigate the risk factors for diabetic severe neuropathy independent of glycemic control and duration of diabetes, erythrocyte aldose reductase was determined in 43 non-insulin-dependent diabetic patients by a two-site ELISA using recombinant human aldose reductase. Among 20 patients with severe neuropathy which was developed within 5 years of diagnosis, the level of erythrocyte aldose reductase protein was significantly higher than that of 23 patients with no or mild neuropathy who had more than 8 years duration of diabetes and prolonged hyperglycemia (11.9+/-5.7 vs. 8.3+/-1.3 ng/mgHb, P < 0.0001). There was a significant stability of the erythrocyte aldose reductase (AR) in the 40 diabetic patients during 1-4 years. The logistic regression analysis revealed that the maximum body mass index (BMI) in the past minus present BMI and the level of erythrocyte aldose reductase protein were the independent risk factors for diabetic severe neuropathy. The measurement of erythrocyte AR level may be useful for predicting severe neuropathy in non-insulin-dependent diabetes mellitus (NIDDM) patients.


[Etiologic analysis of globozoospermia: a 1-case report].
To investigate the etiologic factors of globozoospermia. Routine semen analysis, sperm DNA special staining and chromosomal karyotype detection of peripherical blood lymphocytes were performed in a globozoospermia patient. Round-headed spermatozoa were lack of acrosome and the acrosin activity was low. Meanwhile, there was an additional band located in the Y chromosomal short arm. Lack of acrosome, low acrosin activity and abnormality of chromosome may be the main reasons for globozoospermia.


Molecularly Imprinted Cryogels for the Selective Adsorption of Salicylic Acid.
In this study, molecularly imprinted cryogels were fabricated for selective adsorption of salicylic acid. Cryogelation was performed at - 20 °C using a cationic monomer N,N-dimethylaminoethyl methacrylate as a functional monomer for salicylic acid. The morphology, swelling behaviors, and chemical structures of the cryogels were investigated. The general structure and porosities of cryogels were compared with the traditional hydrogels using field emission scanning electron microscopy (FE-SEM). The adsorption performance of cryogels toward salicylic acid was studied to investigate the optimal adsorption conditions. Adsorption capacity of the imprinted cryogels was 1.95 and 7.51 times higher than those of non-imprinted and bare PHEMA cryogels, respectively, due to the specific binding sites toward salicylic acid. Molecularly imprinted cryogels exhibited significant stability and reusability by keeping more than 85% of their adsorption capacity after ten regeneration cycles. Considering the fabrication process, adsorption capacity, selectivity, and reusability of the imprinted cryogels, these new materials could be utilized as a promising alternative for selective adsorption of drug molecules.


Bleomycin-induced chromosomal aberrations and sister chromatid exchanges in Down lymphocyte cultures.
Peripheral blood lymphocytes from three patients with Down syndrome (DS; trisomy 21; aged 5-6 years) and three age-matched control children were studied for the induction of chromosomal aberrations and sister chromatid exchanges (SCEs). Cells in G0 were exposed to bleomycin (20-100 micrograms/ml) for 3 h, and then cultured in medium containing 5-bromodeoxyuridine and phytohemagglutinin for 66 h. By the sister chromatid differential staining method, chromosome analyses were performed on metaphase cells that had divided one, two, or three or more times after treatment. The results indicate that DS cells exposed to bleomycin are hypersensitive to the production of dicentric and ring chromosomes compared to normal cells. Bleomycin also led to a dose-related increase in the frequency of SCEs, but no difference was found between the SCE frequencies in DS or normal lymphocytes exposed to bleomycin.


Characteristics of Ethical Leadership: Themes Identification Through Convergent Parallel Mixed Method Design From the Pakistan Context.
The current approaches in identifying the characteristics of ethical leadership proceed mainly from a Western perspective based on virtue-driven moral philosophy (i.e., relativism) and frequently ignoring the Asian perspective of morality based on idealism. This study aimed to conduct parallel analysis in convergent design by using qualitative and quantitative methods to extract person-driven ethical leadership themes by considering the Asian context. Using the hypothetico-deductive method, 13 themes were extracted altogether, out of which 4 are new context-driven themes (i.e., altruism, encouragement, collective good, and spiritual transcendence as the emerging themes of ethical leadership in the Asian context).


Assessment of a new microsatellites panel for traceability in Italian inbreed pigs using parentage test
The origin of meat and meat products can be traced by verifying the identity of an offspring from its parents' genotypes. Although there are many microsatellite panels applicable to swine population, efficiency of parental testing decreases when the population consists of consanguineous animals. The aims of the present study were to develop a new microsatellite panel for traceability using parentage test in inbreed pig population and to assess how hybridization can influence the efficiency of parental testing. A new genotyping assay, based on 20‑microsatellite assay, was performed in 304 individuals consisting of related and unrelated animals. The results showed that the microsatellites used in this study display high level of polymorphism ensuring a parentage assignment of 100%. This genotyping panel can be a useful tool to test a 'parent‑to‑fork' traceability system based on 20 microsatellite loci and can overcome technical limitations in inbreed population.


Digital medicine and the curse of dimensionality.
Digital health data are multimodal and high-dimensional. A patient's health state can be characterized by a multitude of signals including medical imaging, clinical variables, genome sequencing, conversations between clinicians and patients, and continuous signals from wearables, among others. This high volume, personalized data stream aggregated over patients' lives has spurred interest in developing new artificial intelligence (AI) models for higher-precision diagnosis, prognosis, and tracking. While the promise of these algorithms is undeniable, their dissemination and adoption have been slow, owing partially to unpredictable AI model performance once deployed in the real world. We posit that one of the rate-limiting factors in developing algorithms that generalize to real-world scenarios is the very attribute that makes the data exciting-their high-dimensional nature. This paper considers how the large number of features in vast digital health data can challenge the development of robust AI models-a phenomenon known as "the curse of dimensionality" in statistical learning theory. We provide an overview of the curse of dimensionality in the context of digital health, demonstrate how it can negatively impact out-of-sample performance, and highlight important considerations for researchers and algorithm designers.


Steroid hormone regulation of C. elegans and Drosophila aging and life history.
In the last two decades it has become clear that hormones and gene mutations in endocrine signaling pathways can exert major effects on lifespan and related life history traits in worms, flies, mice, and other organisms. While most of this research has focused on insulin/insulin-like growth factor-1 signaling, a peptide hormone pathway, recent work has shown that also lipophilic hormones play an important role in modulating lifespan and other life history traits. Here we review how steroid hormones, a particular group of lipophilic hormones, affect life history traits in the nematode worm (Caenorhabditis elegans) and the fruit fly (Drosophila melanogaster), with a particular focus on longevity. Interestingly, a comparison suggests that parallel endocrine principles might be at work in worms and flies in these species and that steroid hormones interact with the gonad to affect lifespan.


Serum antibody immunoreactivity to equine zona protein after SpayVac vaccination.
Immunocontraception with porcine ZP (pZP) can be an effective means of fertility control in feral horses. Previous studies suggest that antibodies produced after pZP vaccination may both inhibit fertilization and cause follicular dysgenesis. Zonastat-H, PZP-22, and SpayVac are three pZP vaccines proposed for use in horses. Although all these vaccines contain the pZP antigen, variations in antigen preparation and vaccine formulation lead to differences in antigenic properties among them. Likewise, despite numerous efficacy and safety studies of Zonastat-H and PZP-22, the contraceptive mechanisms of SpayVac remain unclear. The preparation of pZP for SpayVac is thought to include more nonzona proteins, making it less pure than the other two vaccines. This may result in increased antigenicity of the vaccine. We therefore investigated the immunoreactivity of serum antibodies from SpayVac-vaccinated mares to equine zona protein. Western blot analyses revealed an immunoreactivity of these antibodies to protein isolated from mature equine oocytes, ZP, follicular tissues, and ovarian tissues. Immunohistochemical analyses were used to locate the binding of serum antibodies to the ZP of immature oocytes in ovarian stromal tissue. We also found serum antibodies from SpayVac-treated mares to be predominantly specific for zona protein 3. Collectively, our results suggest a model where serum antibodies produced in response to SpayVac vaccination are immunoreactive to equine zona protein in vitro. Our study lends insight into the contraceptive mechanisms underlying the infertility observed after SpayVac vaccination.


Gfi-1 promotes proliferation of human cervical carcinoma via targeting of FBW7 ubiquitin ligase expression.
The independent growth factor 1 (Gfi-1) is a transcription factor essential for several diverse hematopoietic functions and developments. However, the role and molecular mechanism of Gfi-1 in the development and progression of cervical cancer remains unclear. The present study investigates the relation of expression of Gfi-1 with prognoses in patients with cervical cancer. We used Western blot and reverse transcription polymerase chain reaction (RT-PCR) and the inhibition of proliferation and metastasis of cervical cancer cells in vitro. This study confirms that the expression of Gfi-1 in cervical cancer tissues was higher than that in adjacent normal tissues. The level of Gfi-1 mRNA in human cervical cancer tissues was significantly higher than that in normal tissues adjacent to cancer. Furthermore, overexpression of Gfi-1 promoted cell proliferation, colony formation, and migration of cervical cancer cells. The increased expression of Gfi-1 promotes the proliferation of cervical cancer cells targeting the tumor suppressor F-box and WD repeat domain containing 7 (FBW7). Clinically, our data suggest that overexpression of Gfi-1 is associated with poor prognosis in patients with cervical cancer. In a tumor xenograft model, knockdown of Gfi-1 inhibited the tumor growth of Hela cells in vivo. Our results reveal that Gfi-1 plays an important role in cervical cancer and Gfi-1/FBW7 axis serves as a potential therapeutic target for cervical cancer.


The efficacy of GABAergic precursor cells transplantation in alleviating neuropathic pain in animal models: a systematic review and meta-analysis.
Current therapies are quite unsuccessful in the management of neuropathic pain. Therefore, considering the inhibitory characteristics of GABA mediators, the present systematic review and meta-analysis aimed to determine the efficacy of GABAergic neural precursor cells on neuropathic pain management. Search was conducted on Medline, Embase, Scopus, and Web of Science databases. A search strategy was designed based on the keywords related to GABAergic cells combined with neuropathic pain. The outcomes were allodynia and hyperalgesia. The results were reported as a pooled standardized mean difference (SMD) with a 95% confidence interval (95% CI). Data of 13 studies were analyzed in the present meta-analysis. The results showed that administration of GABAergic cells improved allodynia (SMD = 1.79; 95% CI: 0.87, 271; P < 0.001) and hyperalgesia (SMD = 1.29; 95% CI: 0.26, 2.32; P = 0.019). Moreover, the analyses demonstrated that the efficacy of GABAergic cells in the management of allodynia and hyperalgesia is only observed in rats. Also, only genetically modified cells are effective in improving both of allodynia, and hyperalgesia. A moderate level of pre-clinical evidence showed that transplantation of genetically-modified GABAergic cells is effective in the management of neuropathic pain. However, it seems that the transplantation efficacy of these cells is only statistically significant in improving pain symptoms in rats. Hence, caution should be exercised regarding the generalizability and the translation of the findings from rats and mice studies to large animal studies and clinical trials.


The metabolic activation of 7-methylbenz(a)anthracene in mouse skin.
The metabolism of 7-methylbenz(a)anthracene by rat-liver preparations and by mouse skin has been studied using a combination of thin-layer and high pressure liquid chromatography and all five possible trans-dihydrodiols have been detected as metabolites but in different proportions. The roles of these dihydrodiols and of the related vicinal diol-epoxides in the metabolic activation of 7-methylbenz(a)anthracene in mouse skin has been studied using Sephadex LH-20 column chromatography. The results show that the hydrocarbon-nucleic acid products formed in mouse skin in vivo most probably arise from 3,4-dihydro-3,4-dihydroxy-7-methylbenz(a)anthracene 1,2-oxide which, on the basis of this and other evidence, appears to be the reactive intermediate involved in the metabolic activation of 7-methylbenz(a)anthracene in this tissue.


The topographical organization of motor processing: An ALE meta-analysis on six action domains and the relevance of Broca's region.
Action is a cover term used to refer to a large set of motor processes differing in domain specificities (e.g. execution or observation). Here we review neuroimaging evidence on action processing (N = 416; Subjects = 5912) using quantitative Activation Likelihood Estimation (ALE) and Meta-Analytic Connectivity Modeling (MACM) approaches to delineate the functional specificities of six domains: (1) Action Execution, (2) Action Imitation, (3) Motor Imagery, (4) Action Observation, (5) Motor Learning, (6) Motor Preparation. Our results show distinct functional patterns for the different domains with convergence in posterior BA44 (pBA44) for execution, imitation and imagery processing. The functional connectivity network seeding in the motor-based localized cluster of pBA44 differs from the connectivity network seeding in the (language-related) anterior BA44. The two networks implement distinct cognitive functions. We propose that the motor-related network encompassing pBA44 is recruited when processing movements requiring a mental representation of the action itself.


Genome-wide analysis reveals downregulation of miR-379/miR-656 cluster in human cancers.
MicroRNAs (miRNAs) are non-uniformly distributed in genomes and ~30% of the miRNAs in the human genome are clustered. In this study we have focused on the imprinted miRNA cluster miR-379/miR-656 on 14q32.31 (hereafter C14) to test their coordinated function. We have analyzed expression profile of >1000 human miRNAs in >1400 samples representing seven different human tissue types obtained from cancer patients along with matched and unmatched controls. We found 68% of the miRNAs in this cluster to be significantly downregulated in glioblastoma multiforme (GBM), 61% downregulated in kidney renal clear cell carcinoma (KIRC), 46% in breast invasive carcinoma (BRCA) and 14% in ovarian serous cystadenocarcinoma (OV). On a genome-wide scale C14 miRNAs accounted for 12-30% of the total downregulated miRNAs in different cancers. Pathway enrichment for the predicted targets of C14 miRNA was significant for cancer pathways, especially Glioma (p< 3.77x10⁻⁶, FDR<0.005). The observed downregulation was confirmed in GBM patients by real-time PCR, where 79% of C14 miRNAs (34/43) showed downregulation. In GBM samples, hypermethylation at C14 locus (p<0.003) and downregulation of MEF2, a crucial transcription factor for the cluster was observed which likely contribute to the observed downregulation of the entire miRNA cluster. We provide compelling evidence that the entire C14 miRNA cluster is a tumor suppressor locus involved in multiple cancers, especially in GBM, and points toward a general mechanism of coordinated function for clustered miRNAs.


A novel class of sugar-based ether-linked-dispirooxindolo-pyrrolidines/pyrrolizidines through [3+2]-cycloaddition of azomethine ylides.
An efficient one-pot synthesis of novel sugar based dispirooxindolo-pyrrolidines/pyrrolizidines has been accomplished by a [3+2]-cycloaddition. This method utilizes an azomethine ylide derived from isatin and sarcosine/l-proline, with an ether linked α-,β-unsaturated-β-C-glycosidic ketones as the dipolarophile. All these sugar-based heterocyclics were characterized by NMR ((1)H and (13)C) and elemental analysis. Antimicrobial studies of sugar based dispiroheterocyclic compound 10 shows excellent activity against different microbes.


Ethnomedicinal knowledge, belief and self-reported practice of local inhabitants on traditional antimalarial plants and phytotherapy.
This paper reveals the trend of knowledge and practice of traditional antimalarial plants (TAPs) to prevent/treat malaria. Stratified, systematic random sampling was adopted. The ethnomedicinal survey was conducted between January and March 2011 by involving the selected 371 household members on knowledge and practice of TAPs by administering a pre-tested questionnaire. Overall, 54.4% respondents had adequate awareness and usage custom of TAPs and 16 types of plant have been most commonly known and employed by the respondents to prevent/treat malaria. Leaves (57.2%) were most commonly used plant part to prepare traditional antimalarial phytotherapy remedies. Decoction was one of the most commonly used methods to administer TAPs. The chi-square test result revealed that a significant association is found between the usage custom of traditional antimalarial plants and gender (P-value=0.0282), age (P-value=0.0024), educational status (P-value=0.0295), and monthly income (P-value=0.0001), although not with the ethnicity (P-value=0.7933) of the respondents. TAPs usage is an integral part of the tradition and custom of the Ethiopians. However, nearly half of the respondents have had lack of awareness about TAPs and majority of them are reluctant in exercising either due to its ineffectiveness or its bitter taste. Therefore, further laboratory-based research is extremely imperative to identify their antiplasmodial activity and bioactive molecules which could pave the way to formulate the novel affordable as well as accessible potent antimalarials in the near future.


Serotonin 5HT2 receptor imaging in the human brain using positron emission tomography and a new radioligand, [18F]altanserin: results in young normal controls.
Changes in serotonin-2 receptors have been demonstrated in brain autopsy material from patients with various neurodegenerative and affective disorders. It would be desirable to locate a ligand for the study of these receptors in vivo with positron emission tomography (PET). Altanserin is a 4-benzoylpiperidine derivative with a high affinity and selectivity for S2 receptors in vitro. Dynamic PET studies were carried out in nine normal volunteers with high-specific activity (376-1,680 mCi/mumol) [18F]altanserin. Arterial blood samples were obtained and the plasma time-activity curves were corrected for the presence of labeled metabolites. Thirty minutes after injection, selective retention of the radioligand was observed in cortical areas, while the cerebellum, caudate, and thalamus had low radioactivity levels. Specific binding reached a plateau between 30 and 65 min postinjection at 1.8% of the injected dose/L of brain and then decreased, indicating the reversibility of the binding. The total/nonspecific binding ratio reached 2.6 for times between 50 and 70 min postinjection. The graphical analysis proposed by Logan et al. allowed us to estimate the binding potential (Bmax/KD). Pretreatment with ketanserin was given to three volunteers and brain activity remained uniformly low. An additional study in one volunteer showed that [18F]altanserin can be displaced from the receptors by large doses of ketanserin. At the end of the study, unchanged altanserin was 57% of the total plasma activity. These results suggest that [18F]altanserin is selective for S2 receptors in vivo as it is in vitro. They indicate that [18F]altanserin is suitable for imaging and quantifying S2 receptors with PET in humans.


Catastrophic Outcome Following Apparently Trivial Nondominant Transverse Sinus Injury During Resection of a Tentorial Meningioma: Case Report.
A 52-year-old man was admitted to our hospital with symptoms of raised intracranial pressure and cerebellar dysfunction caused by a medium-sized (4 cm in diameter) tentorial meningioma with an infratentorial extension. Preoperative magnetic resonance imaging showed that the tumor indented and possibly partially invaded the adjacent junction of the nondominant transverse and sigmoid sinuses. The contralateral dominant transverse sinus was fully patent. Total surgical removal of the lesion was done through the left retrosigmoid approach. During dissection of the meningioma, some bleeding from the venous sinus was noted, which was easily controlled by packing with hemostatic materials. The initial postoperative period was unremarkable, but approximately 48 h after surgery, acute clinical deterioration caused by hemorrhagic venous infarction of the left cerebellar hemisphere and brain stem developed and necessitated urgent reoperation for the evacuation of hematoma and brain decompression. Thereafter, the patient remained in a prolonged coma with a severe neurological deficit. After several years of extensive neurorehabilitation, he was able to walk with support but had a tracheostomy, required a feeding tube, and voided with a urinary catheter. Such a catastrophic outcome after an apparently trivial nondominant transverse sinus injury during resection of a tentorial meningioma raises the question whether reconstruction of the sinus wall with preservation of its patency might have prevented this complication in our patient.


Biochemical studies on pulmonary response to inhalation of methylene chloride.
Biochemical response to the toxic lung damage induced by inhalation of methylene chloride was studied. Significant increases in protein, hexose, sialic acid, lactate dehydrogenase, acid and alkaline phosphatase content were observed in the cell-free lavage effluents from lungs of exposed rats compared to the control animals. This was interpreted as increased cell damage accompanied by enhanced pulmonary secretions, perhaps of glycoproteins and mucins, as a result of inhalation toxicity.


Data supporting the results of the characterization of the phases and structures appearing during the synthesis process of Ba0.5Sr1.5Zn2-xNixFe12O22 by auto-combustion.
The data presented has to do with identifying the various phases arising during the synthesis of the Y-type hexaferrite series Ba0.5Sr1.5Zn2-xNixFe12O22 by auto-combustion that we deem important for their microstructural and magnetic properties. The data and the related analyses support the research paper "Ni-substitution effect on the properties of Ba0.5Sr1.5Zn2-xNixFe12O22 powders" [1]. Thus, the parameters are presented of the phases appearing after auto-combustion and after the initial annealing at 800 °C, namely, crystal cell and crystallite size. Also, additional data are provided obtained by EDS concerning the Ba:Sr:Zn:Ni:Fe ratio in Ba0.5Sr1.5Zn2-xNixFe12O22 (x = 0.8, 1, 1.5) samples synthesized at 1170 °C for 10 h. The data can be used as a reference in establishing how the phases distinguished during the initial process of auto-combustion affect the Ba0.5Sr1.5Zn2-xNixFe12O22 powders, which are candidates for room-temperature multiferroic materials. The data have not been published previously and are made available to permit critical or further analyses.


Trabecular bone score in patients with liver transplants after 1 year of risedronate treatment.
The aim of this study was to analyse the effect of risedronate on Trabecular Bone Score in liver transplant patients with low bone mass, during 1-year follow-up. In this retrospective cohort study, trabecular bone score (TBS) was calculated from dual X-ray absorptiometry images of the lumbar spine (LS), collected from a prospective randomized open-label 1-year trial performed in liver recipient patients. A total of 89 patients with osteopenia or osteoporosis were randomized to receive RIS plus calcium and vitamin D3 or calcium and vitamin D3. TBS was low in both groups at baseline, 6 and 12 months. Baseline TBS at the LS showed degraded microarchitecture in 22.8% of patients, partially degraded in 40.3%, and normal values in 36.8% of the patients. After 1 year of treatment, no difference in TBS was observed between both groups. No correlations were found between bone mineral density (BMD) and TBS values at any follow-up time point. No relationship was found between BMD, TBS or immunosuppressive drugs with incidental fracture. No significant effect in TBS was observed in liver transplant patients treated with RIS or calcium and vitamin D3 after 1 year of follow-up. In these patients, the clinical usefulness of this new tool should be established.


Inhibition of proliferation-linked signaling cascades with atractylenolide I reduces myofibroblastic phenotype and renal fibrosis.
Renal fibrosis is a frequent axis contributing to the occurrence of end-stage nephropathy. Previously, it has been reported that atractylenolide Ⅰ (ATL-1), a natural compound extracted from Atractylodes macrocephala, has anti-cancer and antioxidant effects. However, the renal anti-fibrotic effects of action remain unclear. In this study, the anti-fibrotic effects of ATL-1 were examined in fibroblasts, tubular epithelial cells (TECs) triggered by TGF-β1 in vitro, and using a unilateral ureteral obstruction (UUO) mouse model in vivo. We found that ATL-1 represses the myofibroblastic phenotype and fibrosis development in UUO kidneys by targeting the fibroblast-myofibroblast differentiation (FMD), as well as epithelial-mesenchymal transition (EMT). The anti-fibrotic effects of ATL-1 were associated with reduced cell growth in the interstitium and tubules, leading to suppression of the proliferation-linked cascades activity consisting of JAK2/STAT3, PI3K/Akt, p38 MAPK, and Wnt/β-catenin pathways. Besides, ATL-1 treatment repressed TGF-β1-triggered FMD and the myofibroblastic phenotype in fibroblasts by antagonizing the activation of proliferation-linked cascades. Likewise, TGF-β1-triggered excessive activation of the proliferation-linked signaling in TECs triggered EMT. The myofibroblastic phenotype was repressed by ATL-1. The anti-fibrotic and anti-proliferative effects of ATL-1 were linked to the inactivation of Smad2/3 signaling, partially reversing FMD, as well as EMT and the repression of the myofibroblastic phenotype. Thus, the inhibition of myofibroblastic phenotype and fibrosis development in vivo and in vitro through proliferation-linked cascades of ATL-1 makes it a prospective therapeutic bio-agent to prevent renal fibrosis.


Ovarian cancer ascites induces skeletal muscle wasting in vitro and reflects sarcopenia in patients.
Cachexia-associated skeletal muscle wasting or 'sarcopenia' is highly prevalent in ovarian cancer and contributes to poor outcome. Drivers of cachexia-associated sarcopenia in ovarian cancer remain elusive, underscoring the need for novel and better models to identify tumour factors inducing sarcopenia. We aimed to assess whether factors present in ascites of sarcopenic vs. non-sarcopenic ovarian cancer patients differentially affect protein metabolism in skeletal muscle cells and to determine if these effects are correlated to cachexia-related patient characteristics. Fifteen patients with an ovarian mass and ascites underwent extensive physical screening focusing on cachexia-related parameters. Based on computed tomography-based body composition imaging, six cancer patients were classified as sarcopenic and six were not; three patients with a benign condition served as an additional non-sarcopenic control group. Ascites was collected, and concentrations of cachexia-associated factors were assessed by enzyme-linked immunosorbent assay. Subsequently, ascites was used for in vitro exposure of C2C12 myotubes followed by measurements of protein synthesis and breakdown by radioactive isotope tracing, qPCR-based analysis of atrophy-related gene expression, and NF-κB activity reporter assays. C2C12 protein synthesis was lower after exposure to ascites from sarcopenic patients (sarcopenia 3.1 ± 0.1 nmol/h/mg protein vs. non-sarcopenia 5.5 ± 0.2 nmol/h/mg protein, P < 0.01), and protein breakdown rates tended to be higher (sarcopenia 31.2 ± 5.2% vs. non-sarcopenia 20.9 ± 1.9%, P = 0.08). Ascites did not affect MuRF1, Atrogin-1, or REDD1 expression of C2C12 myotubes, but NF-κB activity was specifically increased in cells exposed to ascites from sarcopenic patients (sarcopenia 2.2 ± 0.4-fold compared with control vs. non-sarcopenia 1.2 ± 0.2-fold compared with control, P = 0.01). Protein synthesis and breakdown correlated with NF-κB activity (rs = -0.60, P = 0.03 and rs = 0.67, P = 0.01, respectively). The skeletal muscle index of the ascites donors was also correlated to both in vitro protein synthesis (rs = 0.70, P = 0.005) and protein breakdown rates (rs = -0.57, P = 0.04). Ascites of sarcopenic ovarian cancer patients induces pronounced skeletal muscle protein metabolism changes in C2C12 cells that correlate with clinical muscle measures of the patient and that are characteristic of cachexia. The use of ascites offers a new experimental tool to study the impact of both tumour-derived and systemic factors in various cachexia model systems, enabling identification of novel drivers of tissue wasting in ovarian cancer.


Influence of rate of induction of hypoxia on the ventilatory response.
1. We have studied the influence of rate of induction of hypoxia on the duration of the phases of the respiratory cycle in seven conscious healthy subjects.2. Nine hypoxic procedures were performed on each subject, comprising three different rates of induction of hypoxia each at three different levels of P(CO2). Ventilation vs. arterial O(2) saturation plots were constructed and patterns of response were analysed in terms of ventilation vs. tidal volume and tidal volume vs. inspiratory duration (T(I)) and vs. expiratory duration (T(E)).3. The highest ventilatory responses were observed during steady-state hypoxia when P(CO2) was held at a level similar to the control mixed venous level; progressive hypoxia at this P(CO2) failed to stimulate ventilation to comparable levels. The ventilatory response was not influenced by rate of induction of hypoxia when P(CO2) was maintained at an end-tidal P(CO2) close to the control level.4. As tidal volume increased in response to hypoxia, T(I) and T(E) shortened. The progressive decrease in cycle duration occurred at all levels of P(CO2) and at all rates of induction of hypoxia, but was most marked under eucapnic conditions during steady state.5. It is concluded that steady-state hypoxia may produce a higher ventilatory response than progressive, non-steady-state hypoxia. Inspiratory time shortens as tidal volume increases irrespective of the rate of induction of hypoxia.


Activities of daily living predict periprocedural myocardial infarction and injury following percutaneous coronary intervention: a cross-sectional study.
In recent years, there has been growing interest in exploring the relationship between activities of daily living (ADL) and cardiovascular diseases. This retrospective cross-sectional study aimed to investigate the association of ADL measured by Barthel index (BI) with periprocedural myocardial infarction (PMI) and injury following percutaneous coronary intervention (PCI). Enrolled patients were stratified into impaired and unimpaired ADL groups according to their BI scores. Logistic regressions were conducted to explore the association of ADL on admission with periprocedural myocardial injury and infarction. Restricted cubic spline (RCS) curve and subgroup analysis were also performed. Totally, 16.4% of patients suffered from PMI; the mean age was 65.8 ± 10.4 years old. RCS analysis showed that the morbidity of periprocedural myocardial infarction and injury showed a downward tendency with increasing BI scores. Multivariable logistic regression analysis demonstrated that impaired ADL was an independent risk factor for periprocedural myocardial infarction (OR = 1.190, 95% CI [1.041, 1.360], P = 0.011) and injury (OR = 1.131, 95% CI [1.017, 1.257], P = 0.023). Subgroup analysis showed that the association between ADL and PMI was founded in several subgroups, while the association between ADL and periprocedural myocardial injury was founded only in BMI ≥ 24 kg/m2 subgroup. Impaired ADL at hospital admission was an independent risk factor for periprocedural myocardial infarction and injury among patients following PCI.


Neural networks of information processing in posttraumatic stress disorder: a functional magnetic resonance imaging study.
Neuroimaging studies report reduced medial prefrontal cortical (particularly anterior cingulate) but enhanced amygdala response to fear signals in posttraumatic Stress Disorder (PTSD). We investigated whether anterior cingulate-amygdala dysregulation in PTSD would generalize to salient, but nonthreat related signals. Individuals with PTSD (n = 14) and age and sex-matched nontraumatized controls (n = 14) completed an auditory oddball paradigm adapted to functional magnetic resonance imaging at a 1.5-T field strength. Controls displayed bilateral activation in ventral anterior cingulate and amygdala networks, and PTSD subjects showed bilateral dorsal anterior cingulate and amygdala activation to targets relative to nontargets. Compared to controls, PTSD subjects showed enhanced responses to targets in the dorsal and rostral anterior cingulate, and left amygdala. Whole-brain analyses confirmed the expected pattern of distributed prefrontal-parietal responses to targets in the oddball task. Greater activity in posterior parietal somatosensory regions was observed in PTSD. Our findings of enhanced anterior cingulate responses in PTSD contrast with reports of reduced activity for threat stimuli, suggesting that the latter may be specific to processing of threat-related content. Activation in rostral and dorsal anterior cingulate, left amygdala and posterior parietal networks in response to salient, nonthreatening stimuli may reflect generalized hypervigilance.


Increasing incidence of contact allergy to the new preservative 1,2-dibromo-2,4-dicyanobutane (methyldibromoglutaronitrile).
In 1991, only a low frequency (0.5%) of sensitization by the newly introduced preservative 1,2-dibromo-2,4-dicyanobutane (synonym: methyldibromoglutaronitrile; trade names: Euxyl K400, Merquat 2200, Tektamer 38) was encountered in the Netherlands. It has subsequently been increasingly used in our country as a preservative in moistened toilet tissues, and also in cosmetics such as body milk, lotions and creams. This prompted us to re-evaluate the frequency of sensitization. From January 1993 until July 1994, all consecutive patients attending for routine patch testing were also tested with 1,2-dibromo-2,4-dicyanobutane (0.1% petrolatum): one of 281 (0.3%), nine of 281 (3.2%) and six of 247 (2.4%) were found to be positive in the first and second halves of 1993 and the first half of 1994, respectively. In at least 12 of 16 patients this allergy was responsible for their presenting complaint (perianal and facial dermatitis). As 12 of 16 patients had intensively used moistened toilet tissues containing this preservative, this habit is presumably the cause of sensitization. Our data indicate that the frequency of sensitization is approaching that of allergy to methyl(chloro)isothiazolinone (Kathon CG; 1993: 2.3% in our hospital), a well-known allergen, which seems to have been partially replaced in cosmetics and toiletries by this new preservative.


Chagas Disease Control-Many Approaches to Prospect.
Chagas disease is an emerging and neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi, estimated to infect 8 to 10 million people worldwide, according to the World Health Organization [...].


Rapamycin enhances docetaxel-induced cytotoxicity in a androgen-independent prostate cancer xenograft model by survivin downregulation.
Docetaxel is a first-line treatment choice in castration-resistant prostate cancer (CRPC). However, the management of CRPC remains an important challenge in oncology. There have been many reports on the effects of rapamycin, which is an inhibitor of the mammalian target of rapamycin (mTOR), in the treatment of carcinogenesis. We assessed the cytotoxic effects of the combination treatment of docetaxel and rapamycin in prostate cancer cells. Furthermore, we examined the relationship between these treatments and survivin, which is a member of the inhibitory apoptosis family. Prostate cancer cells were cultured and treated with docetaxel and rapamycin. The effects on proliferation were evaluated with the MTS assay. In addition, we evaluated the effect on proliferation of the combination treatment induced knockdown of survivin expression by small interfering RNA transfection and docetaxel. Protein expression levels were assayed using western blotting. PC3 cells and xenograft growth in nude mice were used to evaluate the in vivo efficacy of docetaxel and its combination with rapamycin. In vitro and in vivo, the combination of rapamycin with docetaxel resulted in a greater inhibition of proliferation than treatment with rapamycin or docetaxel alone. In addition, in vitro and in vivo, rapamycin decreased basal surviving levels, and cotreatment with docetaxel further decreased these levels. Transfection siRNA against survivin enhanced the cytotoxicity of docetaxel in PC3 cells. The rapamycin-dependent enhancement of the cytotoxic effects of docetaxel was associated with the downregulation of survivin expression. Our results suggest that the combination of docetaxel and rapamycin is a candidate for the improved treatment of advanced prostate cancer.


Practical and convenient synthesis of N-heterocycles: stereoselective cyclization of N-alkenylamides with t-BuOI under neutral conditions.
[Structure: see text] tert-Butyl hypoiodite (t-BuOI) was found to be a powerful reagent for the cyclization of N-alkenylamides leading to a variety of N-heterocycles under extremely mild conditions. When N-alkenylsulfonamides were employed in the reaction, three- to six-membered saturated N-heterocycles were obtained in good to excellent yields with complete stereoselectivity. The method was applicable to the cyclization of alkenylbenzamide derivatives to afford N-, O- or N-, S-heterocycles.


Histopathologic changes in fallopian tubes subsequent to sterilization procedures.
Longitudinal serial and serial step sections of fallopian tubes from more than 100 patients, subsequent to tubal sterilization procedures, were examined. Thirteen of these patients had pregnancies following their sterilizations. The histologic findings at the previous surgical sites were compatible with what would be expected for a normal healing process. Evidence for a unique tubal epithelial process, as suggested by the terms "endosalpingiosis" or "recanalization," was lacking. Likewise, the author found no evidence of tuboperitoneal fistula formation and/or the subsequent occurrence of pregnancy secondary to localized endometriosis. The histologic notations of proximal luminal dilatation, plical attenuation, chronic inflammatory infiltrates with pseudopolyp formation, and the findings of plical thickening in the distal segment of remaining tube after an interruption type of procedure seem to be associated with the length of time from the sterilization procedure. These may be factors related to the apparent reduced success rate, with time, of microsurgical reanastomotic procedures.


Associations among physical activity, body mass index, and health-related quality of life by race/ethnicity in a diverse sample of breast cancer survivors.
Health-related quality of life (HRQOL), body mass index (BMI), and physical activity (PA) levels have all been associated with prognosis following breast cancer and may explain partially the higher mortality for breast cancer in certain racial/ethnic subgroups. In this study, associations between PA, BMI, and HRQOL by race were examined in a sample of breast cancer survivors. Measures of PA, BMI, and HRQOL as well as demographic and medical characteristics of women (N = 3013, 13% nonwhite) who participated in the Women's Healthy Eating and Living Study were assessed at baseline. Analysis of covariance was used to examine the relationship between PA and obesity with HRQOL outcomes. Statistical tests were 2-sided. African American women were less likely to meet guidelines for PA and more likely to be obese than women from other ethnic groups (P < .05). In adjusted models, women who met guidelines for PA reported significantly higher physical health composite (point differences ranged from 10.5 to 21.2 points, all P < .05) and vitality (point differences ranged from 9.9 to 16.5 points, all P < .05) scores than those who did not, regardless of race/ethnicity. Associations between obesity and HRQOL were mixed with fewer associations for Asian American and African American women and stronger associations for whites. Breast cancer survivors from racially and ethnically diverse populations have lower levels of PA and higher rates of obesity that are generally associated with poorer HRQOL. Culturally sensitive PA and weight loss interventions may improve these lifestyle characteristics and result in improved HRQOL.


Identification of potential repurposed drugs for treating endometriosis-associated infertility among women.
Endometriosis is the most common gynecological abnormality seen in 10-15% of women of reproductive age, causing infertility in ∼25% of cases, which calls for treatment. Thus, in this study, we have identified miRNAs and genes involved in endometriosis progression, leading to infertility, by performing gene expression analysis followed by pathway analysis and protein-protein networks study. Further, we have predicted repurposed small molecule drugs that will neutralize the regulatory effect of targeting miRNAs that induce sterility in endometriosis. This study predicted two transcription factors, FOXO1, and CREB1, targeted by miRNAs that can be modulated by the repurposed drugs, BRD-K55473186, and methylstat, respectively, for the treatment of infertility due to endometriosis. The former drug seems better and more effective than the other as it showed stronger binding at the active site of FOXO1. These findings provide the rationale for targeting miRNA-regulated transcriptional regulators controlling several biological processes to treat endometriosis and prevent the recurrence of implantation failure or infertility.


Illness perceptions and self-care behaviours in the first years of living with type 2 diabetes; does the presence of complications matter?
To assess illness perceptions, self-care behaviours and their relationship in recently diagnosed type 2 diabetes mellitus (T2DM) patients with and without diabetes-related complications. Cross-sectional survey among 192 recently diagnosed T2DM patients of whom 23% reported the presence of diabetes-related complications. Illness perceptions and self-care were assessed by the Revised Illness Perception Questionnaire (IPQ-R) and the revised Summary of Diabetes Self-Care Activities (SDSCA) measure. Generally, participating patients perceived T2DM as a chronic, but relatively controllable condition with minor consequences. In the presence of complications, however, T2DM was perceived as more unpredictable with more (serious) consequences and less controllable by self-care or medical treatment. Furthermore, engagement in exercise and foot care was reported more often by patients with complications. Self-care was related to certain illness perception dimensions, and interactions between perceptions and complications were found. T2DM patients in the first years of their illness are often recommended to make lifestyle changes in the absence of noticeable diabetes-related symptoms or complaints. As many T2DM patients do not seem to perceive their condition to be serious and postpone lifestyle changes until diabetes-related complications appear, a major challenge for professionals is to convince asymptomatic patients of the importance of self-care.


Dibasic amino acid interactions with Na+-independent transport system asc in horse erythrocytes. Kinetic evidence of functional and structural homology with Na+-dependent system ASC.
Amino acid transport in horse erythrocytes is regulated by three co-dominant allelomorphic genes coding for high-affinity transport activity (system asc1), low-affinity transport activity (system asc2) and transport-deficiency, respectively. The asc systems are selective for neutral amino acids of intermediate size, but unlike conventional system ASC, do not require Na+ for activity. In the present series of experiments we have used a combined kinetic and genetic approach to establish that dibasic amino acids are also asc substrates, systems asc1 and asc2 representing the only mediated routes of cationic amino acid transport in horse erythrocytes. Both transporters were found to exhibit a strong preference for dibasic amino acids compared with neutral amino acids of similar size. Apparent Km values (mM) for influx via system asc1 were L-lysine (9), L-ornithine (27), L-arginine (27), L-alanine (0.35). Corresponding Vmax estimates (mmol/l cells per h, 37 degrees C) were L-lysine (1.65), L-ornithine (2.15), L-arginine (0.54), L-alanine (1.69). Apparent Km values for L-lysine and L-ornithine influx via system asc2 were approximately 90 and greater than 100 mM, respectively, with Vmax values greater than 2 and greater than 1 mmol/l cells per h, respectively. Apparent Km and Vmax values for L-alanine uptake by system asc2 were 14 mM and 6.90 mmol/l cells per h. In contrast, L-arginine was transported by system asc2 with the same apparent Km as L-alanine (14 mM), but with a 77-fold lower Vmax. This dibasic amino acid was shown to cause cis- and trans-inhibition of system asc2 in a manner analogous to its interaction with system ASC, where the side-chain guanidinium group is considered to occupy the Na+-binding site on the transporter. Concentrations of extracellular L-arginine causing 50% inhibition of zero-trans L-alanine influx and half-maximum inhibition of L-alanine zero-trans efflux were 14 mM (extracellular L-alanine concentration 15 mM) and 3 mM (intracellular L-alanine concentration 15.5 mM), respectively. We interpret these observations as evidence of structural homology between the horse erythrocyte asc transporters and system ASC. Physiologically, intracellular L-arginine may function as an endogenous inhibitor of system asc2 activity.


Cypermethrin Induces the Activation of Rat Primary Microglia and Expression of Inflammatory Proteins.
Cypermethrin activates microglia, which is found to be decisive in neurodegeneration in the experimental rats. While the involvement of microglial activation in toxicant-induced neurodegeneration is reported, the effect of low concentration of cypermethrin on the expression of inflammatory proteins from the rat primary microglia is not yet properly understood. The study intended to delineate the effect of low concentration of cypermethrin on the expression and release of proteins from the microglia. Rat primary microglial cells were treated with cypermethrin to check the expression of inflammatory proteins. Cypermethrin-treated microglia conditioned media and cells were collected to measure the expression and release of inflammatory proteins. Cypermethrin augmented the protein kinase C-δ (PKC-δ), inducible nitric oxide synthase (iNOS), phosphorylated mitogen-activated protein kinase (MAPK) p38 and p42/44, matrix metalloproteinase (MMP)-3, and MMP-9 levels in the cell lysate and tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels in the microglia conditioned media. Pre-treatment with minocycline, a microglial activation inhibitor or rottlerin, a PKC-δ inhibitor, notably reduced the release of TNF-α in the conditioned media and expression of iNOS protein in the microglia. Minocycline reduced the expression of PKC-δ, phosphorylated p38 and p42/44 MAPKs, MMP-3, and MMP-9 proteins in the microglia. While cypermethrin-treated conditioned media induced the toxicity in the rat primary neurons, minocycline or rottlerin reduced the cypermethrin treated microglia conditioned media-induced toxicity. The outcomes of the present study suggest that cypermethrin activates microglia and releases TNF-α and IL-1β as well as up-regulates the expression of PKC-δ, iNOS, phosphorylated p38 and p42/44 MAPKs, MMP-3, and MMP-9 proteins, which could contribute to neurodegeneration.


Level of constraint in revision knee arthroplasty.
Revision total knee arthroplasty (TKA) in the setting of major bone deficiency and/or soft tissue laxity might require increasing levels of constraint to restore knee stability. However, increasing the level of constraint not always correlates with mid-to-long-term satisfactory results. Recently, modular components as tantalum cones and titanium sleeves have been introduced to the market with the goal of obtaining better fixation where bone deficiency is an issue; theoretically, satisfactory meta-diaphyseal fixation can reduce the mechanical stress at the level of the joint line, reducing the need for high levels of constraint. This article reviews the recent literature on the surgical management of the unstable TKA with the goal to propose a modern surgical algorithm for adult reconstruction surgeons.


Lanthanum as an intracellular stain for electron microscopy.
Results obtained after the normal aldehyde fixation of duodenal enterocytes for electron microscopy have been compared with results obtained when 0.1% Malachite Green or 10 mM lanthanum chloride had been added during aldehyde fixation. Sections were examined without further staining, and after counterstaining with lead citrate and uranyl acetate. In unstained sections, lanthanum-treated material showed improved contrast when compared to results from the other two methods. Also, after counterstaining, areas showing excellent contrast were much more frequent and more readily detected in the lanthanum-treated material. In the microvilli of enterocytes fixed in the presence of lanthanum, the plasmalemma-glycocalyx was defined more clearly and the results were more pleasing subjectively. When Malachite Green was present in the fixative, good contrast was observed more frequently than in routinely fixed tissues, but less often than in those treated with lanthanum. It is suggested that the addition of lanthanum chloride or Malachite Green to the fixative may prove useful in many ultrastructural studies.


Treatment of penis carcinoma with interstitially administered iridium; comparison with radium therapy.
The results of treatment of 30 patients with penis carcinoma cured with radium and 18 patients treated with 192Ir were compared. In both groups of patients, core assessed at 3 years was obtained for a similar percentage of patients (89% and 83%). After treatment with 192Ir, radiation necrosis was observed in 16%, and after treatment with radium in 50% of patients. In both groups a similar high percentage of patients (88%) with organs preserved after treatment was found. In group of patients treated with 192Ir the percentage of very good functional effects (69%) was much higher in the group of patients treated with radium (28%).


Affinity capillary electrophoresis in binding study of antithrombin to heparin from different sources.
Heparin, a highly sulfated polydispersed glycosaminoglycan (GAG), is the most widespread clinical anticoagulant; it binds antithrombin III (AT), a member of serine proteinases superfamily, accelerating its antagonist effect on blood coagulation. The binding interaction with AT is an important aspect in characterization of physicochemical properties of GAGs. With the aim at profiling several clinical and experimental heparin batches from different sources (porcine, bovine and ovine mucosa), a quantitative investigation of the binding heparin-AT, was undertaken by means of Affinity Capillary Electrophoresis (ACE). In dynamic-equilibrium ACE, the electrophoretic mobility of the receptor (AT), analyzed in a BGE containing the ligand (the considered GAG), is correlated to ligand concentration and binding constant. In particular, a 20mM sodium phosphate, pH 7.4 buffer (the BGE) was chosen as the neat medium and the experiments were carried out in a highly hydrophilic poly(vinyl alcohol) coated capillary (effective length 8.5 cm). The applied sample, consisting in the receptor AT (0.30 μM) and phenylacetic acid (PAA; 10.0 μM) used as a reference compound, was electrophoresed at each of the studied concentration levels of the ligand (heparin samples, 0.30-10.0 × 10(-7)M; heparan sulfate, 0.35-8.0 × 10(-5)M) supplemented to the BGE. The migration time ratio of PAA to AT was assumed as the chemical response to be correlated to the ligand concentration and the binding constant estimation was based on the application of a nonlinear regression method (rectangular hyperbola). Under these conditions, a number of heparin samples were analyzed and their binding constants (Kd) were found within 14.2 and 56.1 nM (SD ≤ ± 2.0; n=3; coefficient of determination r(2) ≥ 0.96). The good correlation of Kd values to the in-vitro activity (anti-factor Xa and anti-factor IIa), confirmed that the affinity for the target AT is an important feature of heparin samples and could be included among their physico-chemical characteristics.


The Missing Piece? A Case for Microglia's Prominent Role in the Therapeutic Action of Anesthetics, Ketamine, and Psychedelics.
There is much excitement surrounding recent research of promising, mechanistically novel psychotherapeutics - psychedelic, anesthetic, and dissociative agents - as they have demonstrated surprising efficacy in treating central nervous system (CNS) disorders, such as mood disorders and addiction. However, the mechanisms by which these drugs provide such profound psychological benefits are still to be fully elucidated. Microglia, the CNS's resident innate immune cells, are emerging as a cellular target for psychiatric disorders because of their critical role in regulating neuroplasticity and the inflammatory environment of the brain. The following paper is a review of recent literature surrounding these neuropharmacological therapies and their demonstrated or hypothesized interactions with microglia. Through investigating the mechanism of action of psychedelics, such as psilocybin and lysergic acid diethylamide, ketamine, and propofol, we demonstrate a largely under-investigated role for microglia in much of the emerging research surrounding these pharmacological agents. Among others, we detail sigma-1 receptors, serotonergic and γ-aminobutyric acid signalling, and tryptophan metabolism as pathways through which these agents modulate microglial phagocytic activity and inflammatory mediator release, inducing their therapeutic effects. The current review includes a discussion on future directions in the field of microglial pharmacology and covers bidirectional implications of microglia and these novel pharmacological agents in aging and age-related disease, glial cell heterogeneity, and state-of-the-art methodologies in microglial research.


Hop1 and the meiotic DNA-damage response.
During the DNA-damage response, adaptor proteins mediate signaling between the PI3K-like sensor kinases, ATM and ATR, and serine/threonine effector kinases. Carballo et al. (2008) now show that the chromosomal protein Hop1 mediates PI3K-like kinase signaling during the repair of DNA double-strand breaks (DSBs) in meiosis.


Somatotopic mapping of chordotonal organ neurons in a primitive ensiferan, the New Zealand tree weta Hemideina femorata: I. femoral chordotonal organ.
The femoral chordotonal organ (FCO) in orthopteran insects comprises several hundred sensory neurons, making it one of the most complex insect proprioceptors. The sensory neurons are suspended from the proximal femur, connecting distally to ligaments and to a needle-like apodeme extending from the proximal tibia. They monitor the position and movement of the tibia. To address how this complexity depends on evolutionary status and function, the morphology of the FCO neurons in the primitive orthopteran Hemideina femorata was investigated by staining small populations of identified afferents. As in crickets, the FCOs in all legs of the weta comprise partly fused ventral and dorsal scoloparia, with the former containing two groups of somata, the ventral group (VG) and the dorsal group (DG). However, the dendrites of the DG insert into thin connective tissue attached to the ventral side of the dorsal ligament, forming a "third scoloparium." The VG afferents terminate mainly in the motor association neuropils, whereas afferents from the dorsal scoloparium neurons terminate exclusively in the vibratory neuropil as do the afferents from the subgenual organ, a substrate vibration detector. Several afferents originating in the DG have extensive terminations in the motor association-, vibratory-, and auditory-processing neuropils, indicating lesser functional specialization than in the other groups. The evolutionary development of the FCO is discussed from a comparative viewpoint.


Kidney manifestations of pediatric Sjögren's syndrome.
Approximately 1% of all patients with Sjögren's syndrome (SS) are children. Unlike the adult form, in which sicca syndrome is the main presentation, in children, the most common clinical finding is recurrent enlargement of the salivary glands. In pediatric SS, extraglandular manifestations represent a significant feature and, among these, kidney manifestations are relevant. Kidney involvement is observed in 5-20.5% of children with SS, most frequently tubulointerstitial nephritis. This injury can lead to serious phenotypes, including distal kidney tubular acidosis with the development of severe hypokalemia, which can lead to ECG abnormalities, weakness, and hypokalemic periodic paralysis. Kidney implications in pediatric SS also include nephrolithiasis, nephrocalcinosis, and various types of glomerular damage, which often require immunosuppressive therapies. Laboratory findings are usually comparable to adults, including hyperglobulinemia and high rates of antinuclear antibodies (ANA, 63.6-96.2%), and anti-Ro/SSA (36.4-84.6%). The current classification criteria for SS are inaccurate for the pediatric population, and more specific criteria are needed to improve the diagnostic rate. Due to the rarity of the disease, strong recommendations for treatment are lacking, and several therapeutic strategies have been reported, mostly based on glucocorticoids and disease-modifying antirheumatic drugs, with different outcomes. The aim of this paper is to provide an overview of the kidney implications of pediatric SS based on the latest evidence of the medical literature.


Postoperative Catheter-Directed Thrombolysis Versus Systemic Anticoagulation for Acute Superior Mesenteric Venous Thrombosis.
Little data evaluate catheter-directed thrombolysis (CDT) therapy as a sequential treatment of emergent surgery for patients with acute superior mesenteric venous thrombosis (ASMVT). We compared the outcomes of ASMVT patients receiving CDT via superior mesenteric artery (SMA) with those who had systemic anticoagulation after emergent laparotomy. A single-center retrospective study of ASMVT patients receiving emergent laparotomy from May 2012 to April 2014 was performed. Patients in group I had postoperative systemic anticoagulation and patients in group II underwent postoperative CDT. The demography, etiology, imaging features, clinical outcomes, and complications were compared. Moreover, univariate analysis was performed to identify confounding variables of 30-day mortality. Thirty-two patients (20 males, mean age of 44.9 ± 10.6 years) were included, 17 in group I and 15 in group II. No significant differences of demographic data, etiology, baseline value, and perioperative comorbidity were found. The rate of complete thrombus removal was significantly higher in group II than group I (29.4% vs. 80.0%, P = 0.001). The second-look laparotomy and repeat bowel resection (58.8% vs. 13.3%, P = 0.002) were required in fewer patients in group II (20.0% vs. 70.6%, P = 0.001). The incidence of short-bowel syndrome (SBS; 41.2% vs. 6.7%, P = 0.001) and 30-day mortality (41.2% vs. 6.7%, P = 0.001) were lower in group II. The 1-year survival was also better in group II (52.9% vs. 93.3%, P = 0.014). The incidence of massive abdominal hemorrhage requiring blood transfusion and surgical intervention was 11.8% in group I and 20.0% in group II (P = 0.645). The age, serum D-dimer level, SBS, and postoperative CDT were significant risk factors of 30-day mortality in this study. For ASMVT patients receiving emergent surgery and intraoperative thrombectomy, the algorithm with postoperative CDT via SMA is associated with more favorable clinical outcome compared with systemic anticoagulation.


A continuous sirtuin activity assay without any coupling to enzymatic or chemical reactions.
Sirtuins are NAD(+) dependent lysine deacylases involved in many regulatory processes such as control of metabolic pathways, DNA repair and stress response. Modulators of sirtuin activity are required as tools for uncovering the biological function of these enzymes and as potential therapeutic agents. Systematic discovery of such modulators is hampered by the lack of direct and continuous activity assays. The present study describes a novel continuous assay based on the increase of a fluorescence signal subsequent to sirtuin mediated removal of a fluorescent acyl chain from a modified TNFα-derived peptide. This substrate is well recognized by human sirtuins 1-6 and represents the best sirtuin 2 substrate described so far with a kcat/KM-value of 176 000 M(-1)s(-1). These extraordinary substrate properties allow the first determination of Ki-values for the specific Sirt2 inhibitory peptide S2iL5 (600 nM) and for the quasi-universal sirtuin inhibitor peptide thioxo myristoyl TNFα (80 nM).


Ca2+/calmodulin is critical for brassinosteroid biosynthesis and plant growth.
Brassinosteroids are plant-specific steroid hormones that have an important role in coupling environmental factors, especially light, with plant growth and development. How the endogenous brassinosteroids change in response to environmental stimuli is largely unknown. Ca2+/calmodulin has an essential role in sensing and transducing environmental stimuli. Arabidopsis DWARF1 (DWF1) is responsible for an early step in brassinosteroid biosynthesis that converts 24-methylenecholesterol to campesterol. Here we show that DWF1 is a Ca2+/calmodulin-binding protein and this binding is critical for its function. Molecular genetic analysis using site-directed and deletion mutants revealed that loss of calmodulin binding completely abolished the function of DWF1 in planta, whereas partial loss of calmodulin binding resulted in a partial dwarf phenotype in complementation studies. These results provide direct proof that Ca2+/calmodulin-mediated signalling has a critical role in controlling the function of DWF1. Furthermore, we observed that DWF1 orthologues from other plants have a similar Ca2+/calmodulin-binding domain, implying that Ca2+/calmodulin regulation of DWF1 and its homologues is common in plants. These results raise the possibility of producing size-engineered crops by altering the Ca2+/calmodulin-binding property of their DWF1 orthologues.


Possible involvement of myeloperoxidase in lipid peroxidation.
1. Exposure of liposomes to the MPO-H2O2-Cl- system results in oxidation of lipids. Malondialdehyde and 4-hydroxynonenal are formed. 2. Oxidation of liposomes by stimulated rat neutrophils, assessed by malondialdehyde formation, is inhibited by KCN. This indicates involvement of MPO in the process. 3. The MPO-H2O2 system oxidizes mildly LDL but in the presence of chloride a propagation phase, with a rapid increase of conjugated diene formation, was observed.


Sensitive sequencing method for KRAS mutation detection by Pyrosequencing.
Both benign and malignant tumors represent heterogenous tissue containing tumor cells and non-neoplastic mesenchymal and inflammatory cells. To detect a minority of mutant KRAS alleles among abundant wild-type alleles, we developed a sensitive DNA sequencing assay using Pyrosequencing, ie, nucleotide extension sequencing with an allele quantification capability. We designed our Pyrosequencing assay for use with whole-genome-amplified DNA from paraffin-embedded tissue. Assessing various mixtures of DNA from mutant KRAS cell lines and DNA from a wild-type KRAS cell line, we found that mutation detection rates for Pyrosequencing were superior to dideoxy sequencing. In addition, Pyrosequencing proved superior to dideoxy sequencing in the detection of KRAS mutations from DNA mixtures of paraffin-embedded colon cancer and normal tissue as well as from paraffin-embedded pancreatic cancers. Quantification of mutant alleles by Pyrosequencing was precise and useful for assay validation, monitoring, and quality assurance. Our Pyrosequencing method is simple, robust, and sensitive, with a detection limit of approximately 5% mutant alleles. It is particularly useful for tumors containing abundant non-neoplastic cells. In addition, the applicability of this assay for DNA amplified by whole-genome amplification technique provides an expanded source of DNA for large-scale studies.


Polymerase chain reaction-restriction fragment length polymorphism of ribosomal internal transcribed spacer region analysis on polyacrylamide gel electrophoresis reveals two haplotypes coexisting in Myzus persicae.
Aphids are geographically and economically the most important group of phytophagous insects. They are responsible for various damage on cultivated plants. In our laboratory, we are interested in their ability to transmit phytoviruses such as Rhabdovirus, Cucumovirus, Potyvirus, and Caulimavirus which cause many crop diseases. The present work deals with the study of vectors as analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of internal transcribed spacer (ITS) regions. Electrophoresis of PCR-RFLP products using EcoRV, AluI and DraI restriction enzymes on 10% polyacrylamid gel reveals the coexistence of two different haplotypes in the same individual genome. This result seems to explain some features of aphid evolution.


Biosynthesis of two quercetin O-diglycosides in Escherichia coli.
Various flavonoid glycosides are found in nature, and their biological activities are as variable as their number. In some cases, the sugar moiety attached to the flavonoid modulates its biological activities. Flavonoid glycones are not easily synthesized chemically. Therefore, in this study, we attempted to synthesize quercetin 3-O-glucosyl (1→2) xyloside and quercetin 3-O-glucosyl (1→6) rhamnoside (also called rutin) using two uridine diphosphate-dependent glycosyltransferases (UGTs) in Escherichia coli. To synthesize quercetin 3-O-glucosyl (1→2) xyloside, sequential glycosylation was carried out by regulating the expression time of the two UGTs. AtUGT78D2 was subcloned into a vector controlled by a Tac promoter without a lacI operator, while AtUGT79B1 was subcloned into a vector controlled by a T7 promoter. UDP-xyloside was supplied by concomitantly expressing UDP-glucose dehydrogenase (ugd) and UDP-xyloside synthase (UXS) in the E. coli. Using these strategies, 65.0 mg/L of quercetin 3-O-glucosyl (1→2) xyloside was produced. For the synthesis of rutin, one UGT (BcGT1) was integrated into the E. coli chromosome and the other UGT (Fg2) was expressed in a plasmid along with RHM2 (rhamnose synthase gene 2). After optimization of the initial cell concentration and incubation temperature, 119.8 mg/L of rutin was produced. The strategies used in this study thus show promise for the synthesis of flavonoid diglucosides in E. coli.


Nanoemulsions as Ophthalmic Drug Delivery Systems
Nanoemulsions are liquid-in-liquid dispersion with a droplet size of about 100 nm. They have a transparent appearance, high rate of bioavailability, and increased shelf life. Nanoemulsions mainly consist of oil, water, surfactant, and cosurfactant and can be prepared by high- and low-energy methods. Diluted nanoemulsions are utilized for the delivery of ophthalmic drugs due to their capability to penetrate the deep layers of the ocular structure, provide a sustained release effect, and reduce the frequency of administration and side effects. These nanoemulsions are subjected to certain tests, such as safety, stability, pH profile, rheological studies, and so on. Cationic nanoemulsions are prepared for topical ophthalmic delivery of active ingredients from cationic agents to increase the drug residence time on the ocular surface, reducing their clearance from the ocular surface and improving drug bioavailability. This review article summarizes the main characteristics of nanoemulsions, ophthalmic nanoemulsions, and cationic nanoemulsions and their components, methods of preparation, and the evaluation parameters for ophthalmic nanoemulsions.


Long intracellular retention of 4'-thio-arabinofuranosylcytosine 5'-triphosphate as a critical factor for the anti-solid tumor activity of 4'-thio-arabinofuranosylcytosine.
4'-Thio-arabinofuranosylcytosine (T-araC) is a new cytosine analog, which exhibits excellent antitumor activity against various solid tumor xenografts in mice. T-araC is a structural analog of arabinofuranosylcytosine (araC), which is known to be marginally active against solid tumors. We have continued to study the biochemical pharmacology of T-araC in solid tumor cells to further characterize the mechanism of action of this new agent and to elucidate why these compounds show a profound difference in antitumor activity against solid tumors. AraC was a slightly more potent inhibitor of cell growth than T-araC when cells were continuously exposed to the drugs. However, T-araC was markedly more cytotoxic than araC when high concentrations of the compounds were given for short periods of time. Despite the fact that T-araC is a much poorer substrate, as compared to araC, for deoxycytidine kinase (the rate-limiting step in the formation of the triphosphates), similar intracellular concentrations of T-araC-5'-triphosphate (T-araCTP) and araCTP were formed in cells at these high, pharmacologically relevant concentrations due to similar Vmax's. The major difference in the metabolism of araC and T-araC was that the half-life of T-araCTP was tenfold longer than that of araCTP and much higher levels of T-araCTP were sustained in cells for long durations after exposure to T-araC. Inhibition of cytidine deaminase, deoxycytidylate deaminase, or DNA replication did not affect the half-life of either araCTP or T-araCTP. In addition, the rates of disappearance of the mono- and tri-phosphates of araC and T-araC in crude cell extracts were similar. These results indicated that these enzymes were not rate-limiting in the degradation of the respective triphosphates. However, the rate of phosphorylation of T-araC-5'-monophosphate (T-araCMP) in crude cell extracts was about tenfold greater than that of araCMP. The results of this work suggested that the longer intracellular retention of T-araCTP was responsible for the superior activity of T-araC against solid tumors in vivo, and that the greater activity of T-araCMP as a substrate of UMP/CMP kinase was responsible for the long intracellular half-life of T-araCTP.


Perturbation of 1JC,F Coupling in Carbon-Fluorine Bonds on Coordination to Lewis Acids: A Structural, Spectroscopic, and Computational Study.
A lithiated m-terphenyl ligand bearing fluorine atoms at the ortho positions of the flanking aryl rings was synthesized and characterized using single crystal X-ray diffraction, variable-temperature multinuclear NMR spectroscopy, and computational methods. Changes in 1JC,F on coordination to lithium as a spectroscopic observable parametrizing the strength of the C-F···Li interaction are described, and a general, qualitative relationship between C-F bond lengths, Δ1JC,F values, and the extent of C-F bond activation as a result of Lewis acid coordination is proposed.


Interferon Gamma Mediates Hematopoietic Stem Cell Activation and Niche Relocalization through BST2.
During chronic infection, the inflammatory cytokine interferon gamma (IFNγ) damages hematopoietic stem cells (HSCs) by disrupting quiescence and promoting excessive terminal differentiation. However, the mechanism by which IFNγ hinders HSC quiescence remains undefined. Using intravital 3-dimensional microscopy, we find that IFNγ disrupts the normally close interaction between HSCs and CXCL12-abundant reticular (CAR) cells in the HSC niche. IFNγ stimulation increases expression of the cell surface protein BST2, which we find is required for IFNγ-dependent HSC relocalization and activation. IFNγ stimulation of HSCs increases their E-selectin binding by BST2 and homing to the bone marrow, which depends on E-selectin binding. Upon chronic infection, HSCs from mice lacking BST2 are more quiescent and more resistant to depletion than HSCs from wild-type mice. Overall, this study defines a critical mechanism by which IFNγ promotes niche relocalization and activation in response to inflammatory stimulation and identifies BST2 as a key regulator of HSC quiescence. VIDEO ABSTRACT.


An Instrument for Measuring Critical Appraisal Self-Efficacy in Rheumatology Trainees.
Self-efficacy, the internal belief that one can perform a specific task successfully, influences behavior. To promote critical appraisal of medical literature, rheumatology training programs should foster both competence and self-efficacy for critical appraisal. This study aimed to investigate whether select items from the Clinical Research Appraisal Inventory (CRAI), an instrument measuring clinical research self-efficacy, could be used to measure critical appraisal self-efficacy (CASE). One hundred twenty-five trainees from 33 rheumatology programs were sent a questionnaire that included two sections of the CRAI. Six CRAI items relevant to CASE were identified a priori; responses generated a CASE score (total score range 0-10; higher = greater confidence in one's ability to perform a specific task successfully). CASE scores' internal structure and relation to domain-concordant variables were analyzed. Questionnaires were completed by 112 of 125 (89.6%) trainees. CASE scores ranged from 0.5 to 8.2. The six CRAI items contributing to the CASE score demonstrated high internal consistency (Cronbach's α = 0.95) and unidimensionality. Criterion validity was supported by the findings that participants with higher CASE scores rated their epidemiology and biostatistics understanding higher than that of peers (P < 0.0001) and were more likely to report referring to studies to answer clinical questions (odds ratio 2.47, 95% confidence interval 1.41-4.33; P = 0.002). The correlation of CASE scores with percentage of questions answered correctly was only moderate, supporting discriminant validity. The six-item CASE instrument demonstrated content validity, internal consistency, discriminative capability, and criterion validity, including correlation with self-reported behavior, supporting its potential as a useful measure of critical appraisal self-efficacy.


Nitric oxide synthases II and III and vascular endothelial growth factor are up-regulated in sciatic nerve after spiral cuff electrode implantation.
Nerve cuff electrodes, commonly used in functional electrical stimulation systems, induce local morphological changes that can affect nerve functionality. Nitric oxide (NO) and vascular endothelial growth factor (VEGF) have both neural and vascular effects. We investigated the time-dependent regulation of nitric oxide synthases (NOS) and of VEGF after implantation of spiral cuff electrode around rat sciatic nerve. NOSIII as well as VEGF were up-regulated in both epineurial and endoneurial compartments in cuff-implanted animals along with microvascular changes. Our results suggest that VEGF and NO are implied in morphological and functional alterations occurring in the early time after cuff implantation.


Ocozocoautla de espinosa virus and hemorrhagic fever, Mexico.
Arenavirus RNA was isolated from Mexican deer mice (Peromyscus mexicanus) captured near the site of a 1967 epidemic of hemorrhagic fever in southern Mexico. Analyses of nucleotide and amino acid sequence data indicated that the deer mice were infected with a novel Tacaribe serocomplex virus (proposed name Ocozocoautla de Espinosa virus), which is phylogenetically closely related to Tacaribe serocomplex viruses that cause hemorrhagic fever in humans in South America.


Ligand oligomerization state controls Tie2 receptor trafficking and angiopoietin-2-specific responses.
Angiopoietin 1 (Ang1) is an activating ligand for the endothelial receptor tyrosine kinase Tie2, whereas Ang2 acts as a context-dependent agonist or antagonist that has a destabilizing effect on the vasculature. The molecular mechanisms responsible for the versatile functions of Ang2 are poorly understood. We show here that Ang2, but not Ang1, induces Tie2 translocation to the specific cell-matrix contact sites located at the distal end of focal adhesions. The Ang2-specific Tie2 translocation was associated with distinct Tie2 activation and downstream signals which differed from those of Ang1, and led to impaired cell motility and weak cell-matrix adhesion. We demonstrate that the different oligomeric or multimeric forms of the angiopoietins induce distinct patterns of Tie2 trafficking; the lower oligomerization state of native Ang2 was crucial for the Ang2-specific Tie2 redistribution, whereas multimeric structures of Ang1 and Ang2 induced similar responses. The Ang2-specific Tie2 trafficking to cell-matrix contacts was also dependent on the cell substratum, α2β1-integrin-containing cell-matrix adhesion sites and intact microtubules. Our data indicate that the different subcellular trafficking of Tie2-Ang2 and Tie2-Ang1 complexes generates ligand-specific responses in the angiopoietin-Tie signaling pathway, including modulation of cell-matrix interactions.


SNF1-RELATED KINASE 1 and TARGET OF RAPAMYCIN control light-responsive splicing events and developmental characteristics in etiolated Arabidopsis seedlings.
The kinases SNF1-RELATED KINASE 1 (SnRK1) and TARGET OF RAPAMYCIN (TOR) are central sensors of the energy status, linking this information via diverse regulatory mechanisms to plant development and stress responses. Despite the well-studied functions of SnRK1 and TOR under conditions of limited or ample energy availability, respectively, little is known about the extent to which the 2 sensor systems function and how they are integrated in the same molecular process or physiological context. Here, we demonstrate that both SnRK1 and TOR are required for proper skotomorphogenesis in etiolated Arabidopsis (Arabidopsis thaliana) seedlings, light-induced cotyledon opening, and regular development in light. Furthermore, we identify SnRK1 and TOR as signaling components acting upstream of light- and sugar-regulated alternative splicing events, expanding the known action spectra for these 2 key players in energy signaling. Our findings imply that concurring SnRK1 and TOR activities are required throughout various phases of plant development. Based on the current knowledge and our findings, we hypothesize that turning points in the activities of these sensor kinases, as expected to occur upon illumination of etiolated seedlings, instead of signaling thresholds reflecting the nutritional status may modulate developmental programs in response to altered energy availability.


Can Gender Nouns Influence the Stereotypes of Animals?
Educating about animals in zoos and aquariums poses daily challenges for education teams, who must not only master biological content but also possess communication skills to adapt information for diverse ages and cultures. This research consists of two sequential studies designed to investigate the impact of grammatical genders on animal stereotypes and elicited emotions. In Study 1, four animals were independently chosen based on a set of predefined conditions, which were then used in Study 2. The second study explored whether the presence of grammatical genders in the Portuguese language influenced the perceived stereotypes of four animals (panda bear, giraffe, polar bear, and cheetah) using the Stereotype Content Model framework. For comparison, English-speaking participants were also surveyed, as English lacks grammatical genders. The results demonstrated that grammatical genders influenced the perceived gender, as well as, although only slightly, the warmth, competence, and elicited emotions of some animals. All animals under study were associated with the protective stereotype, regardless of the presence of grammatical gender. This study emphasizes the significance of subtle yet crucial elements in communication, such as grammatical genders, in shaping stereotypes and innate emotional associations concerning animals.


Pain catastrophizing as a predictor for postoperative pain and opiate consumption in total joint arthroplasty patients.
Pain catastrophizing has been suggested as a prospective risk factor for poor postoperative pain outcomes in total joint arthroplasty (TJA). However, results from the previous studies have been mixed and have not controlled for postoperative opiate analgesic intake. This study investigates pain catastrophizing and postoperative pain intensity in TJA patients, adjusting for analgesic intake. We hypothesized that "pain catastrophizers" would exhibit higher pain scores and increased analgesic requirements postoperatively. In this prospective cohort study, patients were defined as catastrophizers (PCS > 30), or non-catastrophizers (PCS ≤ 30). The primary outcome was the visual analog scale (VAS) pain score at 3-month follow-up. Secondary outcomes included length of stay (LOS) for the index hospitalization, total daily opiate analgesic intake, and VAS pain scores on postoperative days 0, 1, 2, and 3 through discharge. Multivariable regression was used to control for total daily morphine equivalent dose consumed during the stay in addition to other clinical and demographic factors. There were 87 patients in the "non-catastrophizing" and 36 in the "catastrophizing" groups. There was no clinically significant difference in VAS pain scores between groups at 3-month follow-up. Patients with a length of stay (LOS) ≥ 3 postoperative days differed in VAS pain scores ("non-catastrophizers" = 5.08 vs. "catastrophizers" = 7.13; p = 0.002) and were 2.4 times more likely to be catastrophizers than non-catastrophizers (p = 0.042). There were no differences in the remaining secondary outcomes. The pain catastrophizing scale is a poor predictor of postoperative pain at 3-month follow-up. However, it may be a risk factor for increased LOS.


[Analysis of gene variant in a Chinese pedigree with preaxial polydactyly].
To analyze the pathogenic variant of preaxial polydactyly in a Chinese Han pedigree and identify the cause of polydactyly. The peripheral blood DNA of the proband and her parents was extracted. The polydactyly-related genes were detected by trio whole exome sequencing, and the suspected pathogenic gene was screened out. Sanger sequencing was applied to other members of the pedigree. The results of gene sequencing showed that the LMBR1 gene had a heterozygous variant of c.423+4909(IVS5)C>T in 6 patients of the pedigree. The same variant was not detected in family members with normal phenotype. Based on the ACMG guidelines, c.423+4909(IVS5)C>T of the LMBR1 gene was predicted to be pathogenic (PM1+PM2+PP1-S(PS)+PP4+PP5). The heterozygous C>T variant at position 4909 of intron 5 of the LMBR1 gene probably underlies the disease in this pedigree.


Phylogeny of the stapes prosthesis.
To create the first ever stapes prosthesis phylogenetic tree for the evolution of the stapes prosthesis. Retrospective literature review, personal interviews. University Medical Center. Not applicable. Not applicable. Construction of a stapes prosthesis phylogenetic tree with branches capable of including all stapes prostheses. One hundred five different stapes prostheses were reviewed, starting with the first-ever prosthesis used in the first stapedectomy and continuing up to the present time. The stapes prosthesis family tree contains 4 main branches that allow for categorization of all the commercial prostheses currently used. Many examples of atavistic prostheses, single surgeon use, and dead-end characteristics exist. An overview of the complicated phylogenetic tree for stapes prostheses gives great perspective to the history of stapedectomy and insights into many characteristics that are useful for designing new stapes prostheses.