Clinical predictors of length of stay in adults with congenital heart disease.
Length of stay (LOS) is a major driver of inpatient care costs. To date, few studies have investigated risk factors associated with increased LOS in patients with adult congenital heart disease (ACHD). In the present work, we sought to address this knowledge gap. We conducted an analysis of the State Inpatient Databases from Arkansas, California, Florida, Hawaii, Nebraska and New York. We analysed data on admissions in patients with ACHD and constructed a series of hierarchical regression models to identify the clinical factors having the greatest effects on LOS. We identified 99 103 inpatient hospitalisations meeting criteria for inclusion. Diagnoses associated with the longest LOS were septicaemia (LOS=14.2 days in patients atrial septal defect, and 11.7 days among all other ACHD) and pericarditis, endocarditis and myocarditis (LOS=13.6 days and 10.0 days, respectively). When separated by underlying anatomy, the variables most consistently associated with longer LOS were bacterial infection, complications of surgeries or medical care, acute renal disease and anaemia. In the present study, we identified risk factors associated with longer LOS in ACHD. These data may be used to identify at-risk patients for targeted intervention to decrease LOS and thereby cost.


Renal haemodynamics and coronary atherosclerotic burden are associated in patients with hypertension and mild coronary artery disease.
Intrarenal hemodynamic alterations are independent predictors of cardiovascular events in different populations. It has been hypothesized that there is an association between renal hemodynamics and coronary atherosclerotic burden in patients with hypertension. Therefore, the present study examined the associations between renal hemodynamics, coronary atherosclerotic burden and carotid atherosclerotic disease. A total of 130 patients with hypertension aged between 30-80 years who had been referred for an elective coronary angiography were enrolled in the present study. A duplex ultrasound of the intrarenal vasculature was performed to evaluate the resistive index (RI), pulsatility index (PI) and acceleration time (AT). The carotid intima-media thickness was additionally assessed. A coronary angiography was performed to detect the atherosclerotic burden using the Gensini Score (GS). Based on the GS values, subjects were divided into quintiles (I: ≤9; II: 9-17; III: 17-30; IV: 30-44; and V: GS >44) as well as in subjects with mild (GS ≤30) or severe coronary disease (GS >30). A weak significant difference in PI was identified among quintiles (P=0.041), whereas, RI and AT did not differ significantly. PI was associated with GS in the group with low coronary atherosclerotic burden (GS ≤30; P=0.047), whereas, no association was detected in subjects with GS >30. This association remained following adjustment for age and left ventricular ejection fraction (P=0.025). In conclusion, renal vascular alterations were associated with coronary atherosclerotic burden in patients with hypertension with mild coronary disease.


Alzheimer's presenilin 1 mutations impair kinesin-based axonal transport.
Several lines of evidence indicate that alterations in axonal transport play a critical role in Alzheimer's disease (AD) neuropathology, but the molecular mechanisms that control this process are not understood fully. Recent work indicates that presenilin 1 (PS1) interacts with glycogen synthase kinase 3beta (GSK3beta). In vivo, GSK3beta phosphorylates kinesin light chains (KLC) and causes the release of kinesin-I from membrane-bound organelles (MBOs), leading to a reduction in kinesin-I driven motility (Morfini et al., 2002b). To characterize a potential role for PS1 in the regulation of kinesin-based axonal transport, we used PS1-/- and PS1 knock-inM146V (KIM146V) mice and cultured cells. We show that relative levels of GSK3beta activity were increased in cells either in the presence of mutant PS1 or in the absence of PS1 (PS1-/-). Concomitant with increased GSK3beta activity, relative levels of KLC phosphorylation were increased, and the amount of kinesin-I bound to MBOs was reduced. Consistent with a deficit in kinesin-I-mediated fast axonal transport, densities of synaptophysin- and syntaxin-I-containing vesicles and mitochondria were reduced in neuritic processes of KIM146V hippocampal neurons. Similarly, we found reduced levels of PS1, amyloid precursor protein, and synaptophysin in sciatic nerves of KIM146V mice. Thus PS1 appears to modulate GSK3beta activity and the release of kinesin-I from MBOs at sites of vesicle delivery and membrane insertion. These findings suggest that mutations in PS1 may compromise neuronal function by affecting GSK-3 activity and kinesin-I-based motility.


Crusading for the forgotten: Dr. Peter Bryce, public health, and prairie native residential schools.
This article examines the rise of the public health movement and its impact on native affairs in Canada at the turn of the century. It focuses on the efforts of Peter Bryce, Chief Medical Officer for the Departments of the Interior and Indian Affairs, to improve conditions in native schools and on reserves. In 1907 Bryce released his controversial Report on the Indian Schools of Manitoba and the North West Territories, revealing that 24 percent of all native residential school students had died of tuberculosis. Believing firmly that the state was responsible for promoting the health and welfare of its people, Bryce insisted that the federal government address this high death rate. His tireless crusade on behalf of the native population demonstrated the extent to which native affairs were influenced by the broader social, political, and economic agendas of the day, and anticipated the federal government's changing role in social services following World War I.


Distinct spatial Ca2+ signatures selectively activate different NFAT transcription factor isoforms.
Protein isoforms are widely expressed in biological systems. How isoforms that co-exist within the same sub-cellular domain are differentially activated remains unclear. Here, we compare the regulatory mechanism of two closely related transcription factor isoforms, NFAT1 and NFAT4, that migrate from the cytoplasm to the nucleus following the increase in intracellular Ca(2+) that accompanies the opening of store-operated Orai1/CRAC channels. We demonstrate that NFAT1 has a private line of communication with Orai1, activating in response to Ca(2+) microdomains near the open channels. By contrast, NFAT4 stimulation requires both local Ca(2+) entry and a nuclear Ca(2+) rise. We mapped differences in nuclear location to amino acids within the SP-3 motif of the NFAT regulatory domain. The different Ca(2+) dependencies enable agonists to recruit different isoform combinations as stimulus strength increases. Our study uncovers a mechanism whereby co-existing cytoplasmic transcription factor isoforms are differentially activated by distinct sub-cellular Ca(2+) signals.


The timing of the formation and usage of replicase clusters in S-phase nuclei of human diploid fibroblasts.
The sites of nascent DNA synthesis were compared with the distribution of the proliferating cell nuclear antigen (PCNA) in S-phase nuclei of human diploid fibroblasts (HDF) by two in vitro techniques. Firstly, proliferating fibroblasts growing in culture that had been synchronised at S-phase were microinjected with the thymidine analogue biotin-11-dUTP. The sites of incorporation of biotin into injected cells were compared with the distribution of PCNA by indirect immunofluorescence microscopy and laser scanning confocal microscopy (LSCM). In common with other studies, a progression of patterns for both biotin incorporation and PCNA localisation was observed. However, we did not always observe coincidence in these patterns, the pattern of biotin incorporation often resembling the expected, preceding distribution of PCNA. In nuclei in which the pattern of biotin incorporation appeared to be identical to the distribution of PCNA, LSCM revealed that not all of the sites of PCNA immunofluorescence were incorporating biotin at the same time. Secondly, nuclei which had been isolated from quiescent cultures of HDF were innoculated into cell-free extracts of Xenopus eggs which support DNA replication in vitro. Following innoculation into these extracts DNA replication was initiated in each nucleus. The sites of DNA synthesis were detected by biotin-11-dUTP incorporation and compared with the distribution of PCNA by indirect immunofluorescence. Only a single pattern of biotin incorporation and PCNA distribution was observed. PCNA accumulated at multiple discrete spots some 15 min before any biotin incorporation was observed. When biotin incorporation did occur, LSCM revealed almost complete coincidence between the sites of DNA synthesis and the sites at which PCNA was localised.


Conversion of Amazon rainforest to agriculture alters community traits of methane-cycling organisms.
Land use change is one of the greatest environmental impacts worldwide, especially to tropical forests. The Amazon rainforest has been subject to particularly high rates of land use change, primarily to cattle pasture. A commonly observed response to cattle pasture establishment in the Amazon is the conversion of soil from a methane sink in rainforest, to a methane source in pasture. However, it is not known how the microorganisms that mediate methane flux are altered by land use change. Here, we use the deepest metagenomic sequencing of Amazonian soil to date to investigate differences in methane-cycling microorganisms and their traits across rainforest and cattle pasture soils. We found that methane-cycling microorganisms responded to land use change, with the strongest responses exhibited by methane-consuming, rather than methane-producing, microorganisms. These responses included a reduction in the relative abundance of methanotrophs and a significant decrease in the abundance of genes encoding particulate methane monooxygenase. We also observed compositional changes to methanotroph and methanogen communities as well as changes to methanotroph life history strategies. Our observations suggest that methane-cycling microorganisms are vulnerable to land use change, and this vulnerability may underlie the response of methane flux to land use change in Amazon soils.


Effect of resiquimod 0.01% gel on lesion healing and viral shedding when applied to genital herpes lesions.
Resiquimod, a Toll-like receptor 7/8 agonist developed as a topical treatment to decrease recurrences of anogenital herpes, induces proinflammatory cytokines that may delay lesion healing. Adults with frequently recurring anogenital herpes were randomized within 24 h of onset of a recurrence to vehicle or resiquimod 0.01% gel two times per week for 3 weeks. Subjects underwent daily lesion assessments and sampling for herpes simplex virus DNA PCR for 21 days or until investigator-determined healing of lesion(s). Eighty-two subjects with a mean age of 39 +/- 10.5 years and a median of seven recurrences per year were enrolled in the study. The qualifying recurrence was positive by PCR for herpes simplex virus in 68% of subjects. No difference was observed between the vehicle (39 subjects) and resiquimod (43 subjects) groups with respect to time to healing (median of 7.0 days versus median of 6.5 days, respectively; Cox proportional hazard model ratio of 1.229; 95% confidence interval, 0.778 to 1.942; P = 0.376). The distributions of maximum severity scores for any investigator-assessed local skin signs and for subject-assessed local symptoms were similar between treatment groups (P = 0.807 and P = 0.103, respectively). For subjects with at least one positive PCR result, no difference was observed for time to cessation of viral shedding (median of 7 days versus median of 5 days for vehicle and resiquimod groups, respectively; Cox proportional hazard model ratio of 1.471; 95% confidence interval, 0.786 to 2.754; P = 0.227). Application of resiquimod 0.01% two times per week for 3 weeks did not delay the healing of genital herpes lesions or reduce acute viral shedding.


Suppression of microRNA-31 increases sensitivity to 5-FU at an early stage, and affects cell migration and invasion in HCT-116 colon cancer cells.
MicroRNAs (miRNAs) are endogenously expressed noncoding RNAs with important biological and pathological functions. Although several studies have shown that microRNA-31 (miR-31) is obviously up-regulated in colorectal cancer (CRC), there is no study on the functional roles of miR-31 in CRC. Anti-miR™ miRNA 31 inhibitor (anti-miR-31) is a sequence-specific and chemically modified oligonucleotide to specifically target and knockdown miR-31 molecule. The effect of anti-miR-31 transfection was investigated by real-time PCR. HCT-116p53+/+ and HCT-116p53-/-colon cancer cells were treated by anti-miR-31 with or without 5-fluorouracil (5-FU), cell proliferation was determined by MTT assay; apoptosis was detected by DAPI staining; cell cycle was evaluated by flow cytometry; colony formation, migration and invasion assays were performed to investigate the effect of suppression of miR-31 on the cell lines. Real-time PCR results showed that anti-miR-31 was efficiently introduced into the cells and reduced miR-31 levels to 44.1% in HCT-116p53+/+ and 67.8% in HCT-116p53-/-cell line (p = 0.042 and 0.046). MTT results showed that anti-miR-31 alone had no effect on the proliferation of HCT-116p53+/+ or HCT-116p53-/-. However, when combined with 5-FU, anti-miR-31 inhibited the proliferation of the two cell lines as early as 24 h after exposure to 5-FU (p = 0.038 and 0.044). Suppression of miR-31 caused a reduction of the migratory cells by nearly 50% compared with the negative control in both HCT-116p53+/+ and HCT-116p53-/-(p = 0.040 and 0.001). The invasive ability of the cells were increased by 8-fold in HCT-116p53+/+ and 2-fold in HCT-116p53-/- (p = 0.045 and 0.009). Suppression of miR-31 had no effect on cell cycle and colony formation (p > 0.05). Suppression of miR-31 increases sensitivity to 5-FU at an early stage, and affects cell migration and invasion in HCT-116 colon cancer cells.


Climate change and the decline of a once common bird.
Climate change is predicted to negatively impact wildlife through a variety of mechanisms including retraction of range. We used data from the North American Breeding Bird Survey and regional and global climate indices to examine the effects of climate change on the breeding distribution of the Rusty Blackbird (Euphagus carolinus), a formerly common species that is rapidly declining. We found that the range of the Rusty Blackbird retracted northward by 143 km since the 1960s and that the probability of local extinction was highest at the southern range margin. Furthermore, we found that the mean breeding latitude of the Rusty Blackbird was significant and positively correlated with the Pacific Decadal Oscillation with a lag of six years. Because the annual distribution of the Rusty Blackbird is affected by annual weather patterns produced by the Pacific Decadal Oscillation, our results support the hypothesis that directional climate change over the past 40 years is contributing to the decline of the Rusty Blackbird. Our study is the first to implicate climate change, acting through range retraction, in a major decline of a formerly common bird species.


The effect of catalysts and underlayer metals on the properties of PECVD-grown carbon nanostructures.
The growth behaviors and contact resistances of vertically aligned carbon nanotubes (CNTs) and carbon nanofibers (CNFs) grown on different underlayer metals are investigated. The average diameter, diameter distribution, density, growth rate and contact resistance exhibit strong correlation with the choice of catalyst/underlayer combination. These observations are analyzed in terms of interactions between the catalyst and the underlayer metal. The CNT via test structure has been designed and fabricated to make current-voltage measurements on single CNTs using a nanomanipulator under scanning electron microscopy (SEM) imaging. By analyzing the dependence of measured resistance on CNT diameter, the CNT-metal contact resistance can be extracted. The contact resistances between as-grown CNTs and different underlayer metals are determined. Relationships between contact resistances and various combinations of catalysts and underlayer metals are investigated.


Transplacental transfer of NMDA receptor antibodies in an infant with cortical dysplasia.
A confused and agitated 18-year-old woman presented to the emergency unit with orolingual movements, eye deviation, and a temperature of 38°C. The symptoms had begun 2 weeks prior to the admission when she developed a severe headache associated with pathologic laughing and intermittent episodes of upgaze deviation. A urine pregnancy test was positive and a transvaginal ultrasonography showed a 9-week-old fetus. An MRI of the brain was unremarkable and results of the CSF analysis were also unremarkable apart from a CSF pleocytosis (62 lymphocytes) and slightly elevated protein (55 mg/dL; normal range 0-45 mg/dL). Extensive microbiologic and serologic studies with CSF were all negative. She gradually lost consciousness, experienced respiratory failure, and was intubated. There were semirhythmic movements consisting of complex patterns of mouth opening, chewing, facial grimacing, synchronous flexion-extension, and supination-pronation limb movements, which persisted during the period of unresponsiveness. She also had generalized hyperreflexia, persistent hyperthermia, and a full bladder. Three EEGs showed diffuse slow waves with no epileptic discharges. A diagnosis of anti-NMDA receptor (NMDAR) encephalitis was made on clinical grounds and strongly positive serum NMDAR antibodies.


Cognitive inflexibility after prefrontal serotonin depletion.
Serotonergic dysregulation within the prefrontal cortex (PFC) is implicated in many neuropsychiatric disorders, but the precise role of serotonin within the PFC is poorly understood. Using a serial discrimination reversal paradigm, we showed that upon reversal, selective serotonin depletion of the marmoset PFC produced perseverative responding to the previously rewarded stimulus without any significant effects on either retention of a discrimination learned preoperatively or acquisition of a novel discrimination postoperatively. These results highlight the importance of prefrontal serotonin in behavioral flexibility and are highly relevant to obsessive-compulsive disorder, schizophrenia, and the cognitive sequelae of drug abuse in which perseveration is prominent.


Arsenic induces apoptosis by the lysosomal-mitochondrial pathway in INS-1 cells.
Recently, long term arsenic exposure was considered to be associated with an increased risk of diabetes mellitus. While a relation of cause-and-effect between apoptosis of pancreatic β-cells and arsenic exposure, the precise mechanisms of these events remains unclear. The aim of this study was to explore arsenic-induced pancreatic β-cell apoptosis and the mechanisms of through the possible link between lysosomal and the mitochondrial apoptotic pathway. After exposure to 10 μM of arsenic, the reactive oxygen species (ROS) level was significantly increased at 12 h, while the mitochondrial membrane potential was reduced at 24 h and the lysosomal membrane integrity was disrupted at 48 h. A significant increase in protein expression for cytochrome c was also observed using Western blot analysis after exposure to arsenic for 48 h. To further demonstrate that arsenic reduced the lysosomal membrane integrity, cells pretreated with NH4 Cl and exposed to arsenic harbored a lower fluorescence increase than cells that were only exposed to arsenic. In addition, apoptosis was mesured using Hoechst 33342/PI dual staining by microscopy and annexin V-FITC/propidium iodide dual staining by flow cytometry. The results show an increased uptake of the arsenic dose and the cells changed from dark blue to light blue, karyopyknosis, nuclear chromatin condensation, side set or fracture, and a correlation was found between the number of apoptotic cells and arsenic dose. The result of present study suggest that arsenic may induce pancreatic β-cell apoptosis through activation of the lysosome-mitochondrial pathway.


The role of extracellular and cytoplasmic splice domains of alpha7-integrin in cell adhesion and migration on laminins.
The major laminin-binding integrin of skeletal, smooth, and heart muscle is alpha7beta1-integrin, which is structurally related to alpha6beta1. It occurs in three cytoplasmic splice variants (alpha7A, -B, and -C) and two extracellular forms (X1 and X2) which are developmentally regulated and differentially expressed in skeletal muscle. Previously, we have shown that ectopic expression of the alpha7beta-integrin splice variant in nonmotile HEK293 cells specifically induced cell locomotion on laminin-1 but not on fibronectin. To investigate the specificity and the mechanism of the alpha7-mediated cell motility, we expressed the three alpha7-chain cytoplasmic splice variants, as well as alpha6A- and alpha6B-integrin subunits in HEK293 cells. Here we show that all three alpha7 splice variants (containing the X2 domain), as well as alpha6A and alpha6B, promote cell attachment and stimulate cell motility on laminin-1 and its E8 fragment. Deletion of the cytoplasmic domain (excluding the GFFKR consensus sequence) from alpha7B resulted in a loss of the motility-enhancing effect. On laminin-2/4 (merosin), the predominant isoform in mature skeletal muscle, only alpha7-expressing cells showed enhanced motility, whereas cells transfected with alpha6A and alpha6B neither attached nor migrated on laminin-2. Adhesion of alpha7-expressing cells to both laminin-1 and laminin-2 was specifically inhibited by a new monoclonal antibody (6A11) specific for alpha7. Expression of the two extracellular splice variants alpha7X1 and alpha7X2 in HEK293 cells conferred different motilities on laminin isoforms: Whereas alpha7X2B promoted cell migration on both laminin-1 and laminin-2, alpha7X1B supported motility only on laminin-2 and not on laminin-1, although both X1 and X2 splice variants revealed similar adhesion rates to laminin-1 and -2. Fluorescence-activated cell sorter analysis revealed a dramatic reduction of surface expression of alpha6-integrin subunits after alpha7A or -B transfection; also, surface expression of alpha1-, alpha3-, and alpha5-integrins was significantly reduced. These results demonstrate selective responses of alpha6- and alpha7-integrins and of the alpha7 splice variants to laminin-1 and -2 and indicate differential roles in laminin-controlled cell adhesion and migration.


Three-dimensional bio-printing.
Three-dimensional (3D) printing technology has been widely used in various manufacturing operations including automotive, defence and space industries. 3D printing has the advantages of personalization, flexibility and high resolution, and is therefore becoming increasingly visible in the high-tech fields. Three-dimensional bio-printing technology also holds promise for future use in medical applications. At present 3D bio-printing is mainly used for simulating and reconstructing some hard tissues or for preparing drug-delivery systems in the medical area. The fabrication of 3D structures with living cells and bioactive moieties spatially distributed throughout will be realisable. Fabrication of complex tissues and organs is still at the exploratory stage. This review summarize the development of 3D bio-printing and its potential in medical applications, as well as discussing the current challenges faced by 3D bio-printing.


Production of phytotoxic cationic α-helical antimicrobial peptides in plant cells using inducible promoters.
Synthetic linear antimicrobial peptides with cationic α-helical structures, such as BP100, have potent and specific activities against economically important plant pathogenic bacteria. They are also recognized as valuable therapeutics and preservatives. However, highly active BP100 derivatives are often phytotoxic when expressed at high levels as recombinant peptides in plants. Here we demonstrate that production of recombinant phytotoxic peptides in transgenic plants is possible by strictly limiting transgene expression to certain tissues and conditions, and specifically that minimization of this expression during transformation and regeneration of transgenic plants is essential to obtain viable plant biofactories. On the basis of whole-genome transcriptomic data available online, we identified the Os.hsp82 promoter that fulfilled this requirement and was highly induced in response to heat shock. Using this strategy, we generated transgenic rice lines producing moderate yields of severely phytotoxic BP100 derivatives on exposure to high temperature. In addition, a threshold for gene expression in selected tissues and stages was experimentally established, below which the corresponding promoters should be suitable for driving the expression of recombinant phytotoxic proteins in genetically modified plants. In view of the growing transcriptomics data available, this approach is of interest to assist promoter selection for specific purposes.


Rediscovery of Red Wolf Ghost Alleles in a Canid Population Along the American Gulf Coast.
Rediscovering species once thought to be extinct or on the edge of extinction is rare. Red wolves have been extinct along the American Gulf Coast since 1980, with their last populations found in coastal Louisiana and Texas. We report the rediscovery of red wolf ghost alleles in a canid population on Galveston Island, Texas. We analyzed over 7000 single nucleotide polymorphisms (SNPs) in 60 canid representatives from all legally recognized North American Canis species and two phenotypically ambiguous canids from Galveston Island. We found notably high Bayesian cluster assignments of the Galveston canids to captive red wolves with extensive sharing of red wolf private alleles. Today, the only known extant wild red wolves persist in a reintroduced population in North Carolina, which is dwindling amongst political and taxonomic controversy. Our rediscovery of red wolf ancestry after almost 40 years introduces both positive opportunities for additional conservation action and difficult policy challenges.


Meta-biomaterials.
Meta-biomaterials are designer biomaterials with unusual and even unprecedented properties that primarily originate from their geometrical designs at different (usually smaller) length scales. This concept has been primarily used in the context of orthopedic biomaterials with the ultimate aim of improving the bone tissue regeneration performance of implants and decreasing the risk of implant-associated infections. In this paper, we review the ways though which geometrical design at the macro-, micro-, and nanoscales combined with advanced additive manufacturing techniques (3D printing) could be used to create the unusual properties of meta-biomaterials. Due to their intended applications in orthopedics, metallic and hard polymeric biomaterials have received the most attention in the literature. However, the reviewed concepts are, at least in principle, applicable to a wide range of biomaterials including ceramics and soft polymers. At the macroscale, we discuss the concepts of patient-specific implants, deployable meta-implants, and shape-morphing implants. At the microscale, we introduce the concept of multi-physics meta-biomaterials while also covering the applications of auxetic meta-biomaterials for improving the longevity of orthopedic implants. At the nanoscale, the different aspects of the geometrical design of surface nanopatterns that simultaneously stimulate the osteogenic differentiation of stem cells and kill bacteria are presented. The concept of origami-based meta-biomaterials and the applications of self-folding mechanisms in the fabrication of meta-biomaterials are addressed next. We conclude with a discussion on the available evidence regarding the superior performance of meta-biomaterials and suggest some possible avenues for future research.


Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood.
We aimed to characterise the staphylococcal cassette chromosome mec (SCCmec) type, genetic relatedness, biofilm formation and composition, icaADBC genes detection, icaD expression, and antibiotic susceptibility of planktonic and biofilm cells of Staphylococcus hominis isolates from blood. The study included 67 S. hominis blood isolates. Methicillin resistance was evaluated with the cefoxitin disk test. mecA gene and SCCmec were detected by multiplex PCR. Genetic relatedness was determined by pulsed-field gel electrophoresis. Biofilm formation and composition were evaluated by staining with crystal violet and by detachment assay, respectively; and the biofilm index (BI) was determined. Detection and expression of icaADBC genes were performed by multiplex PCR and real-time PCR, respectively. Antibiotic susceptibilities of planktonic cells (minimum inhibitory concentration, MIC) and biofilm cells (minimum biofilm eradication concentration, MBEC) were determined by the broth dilution method. Eighty-five percent (57/67) of isolates were methicillin resistant and mecA positive. Of the mecA-positive isolates, 66.7% (38/57) carried a new putative SCCmec type. Four clones were detected, with two to five isolates each. Among all isolates, 91% (61/67) were categorised as strong biofilm producers. Biofilm biomass composition was heterogeneous (polysaccharides, proteins and DNA). All isolates presented the icaD gene, and 6.66% (1/15) isolates expressed icaD. This isolate presented the five genes of ica operon. Higher BI and MBEC values than the MIC values were observed for amikacin, vancomycin, linezolid, oxacillin, ciprofloxacin, and chloramphenicol. S. hominis isolates were highly resistant to methicillin and other antimicrobials. Most of the detected SCCmec types were different than those described for S. aureus. Isolates indicated low clonality. The results indicate that S. hominis is a strong biofilm producer with an extracellular matrix with similar composition of proteins, DNA and N-acetylglucosamine; and presents high frequency and low expression of icaD gene. Biofilm production is associated with increased antibiotic resistance.


Low rates of complications for carotid artery stenting are associated with a high clinician volume of carotid artery stenting and aortic endografting but not with a high volume of percutaneous coronary interventions.
Prior studies have demonstrated improved clinical outcomes for surgeons with a high-volume experience with certain open vascular operations. A high-volume experience with carotid artery stenting (CAS) improves clinical outcomes. Moreover, it is not known whether experience with other endovascular procedures, including percutaneous coronary interventions (PCIs), is an adequate substitute for experience with CAS. The goal of this study was to quantify the effect of increasing clinician volume of CAS, endovascular aneurysm repair (EVAR), and thoracic endovascular aortic aneurysm repair (TEVAR), and PCI on the outcomes for CAS. The Nationwide Inpatient Sample was analyzed to identify patients undergoing CAS for the years 2005 to 2009. Clinicians were stratified into tertiles of low-volume, medium-volume, and high-volume groups by annual volume of CAS, EVAR/TEVAR, and PCI. Multiple logistic regression analyses were used to examine the relationship between clinician volume and a composite outcome of the in-hospital stroke and death rate after CAS. Between 2005 and 2009, 56,374 elective CAS procedures were performed nationwide, with a crude in-hospital stroke and death rate of 3.22%. A median of nine CAS procedures (interquartile range, 3-20) were performed annually per clinician. As expected, stroke and death rates for CAS decreased with increasing volume of CAS performed by a clinician (low-volume vs medium-volume vs high-volume: 4.43% vs 2.89% vs 2.27%; P = .0001). Similar patterns were noted between clinicians' volume of EVAR/TEVAR (low-volume vs medium-volume vs high-volume: 4.58% vs 3.18% vs 2.16%; P = .0023). In contrast, increasing PCI volume was not associated with decreased stroke and death rates after CAS (low-volume vs medium-volume vs high-volume: 2.99% vs 3.18% vs 3.55%; P = .35). After adjusting for patient and hospital characteristics, clinician volume of CAS (odds ratio [OR], 0.84; 95% confidence interval [CI], 0.74-0.94; P = .003) and EVAR/TEVAR (OR, 0.85; 95% CI, 0.75-0.97; P = .020) remained significant predictors of stroke and death after CAS, whereas increasing clinician volume of PCI was associated with significantly increasing likelihood of stroke or death after CAS (OR, 1.025; 95% CI, 1.004-1.047; P = .019). The stroke and death rate for CAS to treat carotid stenosis is inversely affected by the number of CAS and EVAR/TEVAR procedures performed by a clinician. In contrast, a high-volume experience with PCI is not associated with improved outcomes after CAS.


Solving the puzzle of metastasis: the evolution of cell migration in neoplasms.
Metastasis represents one of the most clinically important transitions in neoplastic progression. The evolution of metastasis is a puzzle because a metastatic clone is at a disadvantage in competition for space and resources with non-metastatic clones in the primary tumor. Metastatic clones waste some of their reproductive potential on emigrating cells with little chance of establishing metastases. We suggest that resource heterogeneity within primary tumors selects for cell migration, and that cell emigration is a by-product of that selection. We developed an agent-based model to simulate the evolution of neoplastic cell migration. We simulated the essential dynamics of neoangiogenesis and blood vessel occlusion that lead to resource heterogeneity in neoplasms. We observed the probability and speed of cell migration that evolves with changes in parameters that control the degree of spatial and temporal resource heterogeneity. Across a broad range of realistic parameter values, increasing degrees of spatial and temporal heterogeneity select for the evolution of increased cell migration and emigration. We showed that variability in resources within a neoplasm (e.g. oxygen and nutrients provided by angiogenesis) is sufficient to select for cells with high motility. These cells are also more likely to emigrate from the tumor, which is the first step in metastasis and the key to the puzzle of metastasis. Thus, we have identified a novel potential solution to the puzzle of metastasis.


Information needs of United Kingdom primary care clinicians.
The scope of primary care makes knowledge management in this context very challenging, especially as access to information increases and the role of primary care within the NHS expands. This paper reviews the literature on the information needs of primary care clinicians to enable evidence-based decision making. Drawing on the literature, and using a specific example of a clinical informaticist service, some lessons are drawn on the role of information professionals in facilitating evidence-based health care. Literature review. Case study of clinical informaticist service. There are numerous barriers to clinicians engaging with evidence-based practice. However, providing evidence-based information to clinicians' questions in a user-friendly format has the potential to facilitate changes in practice. The successful implementation of information services requires attention to both academic and service dimensions, fostering closer working relationships between clinicians and information professionals.


Analysis of circulating tumor DNA to monitor metastatic breast cancer.
The management of metastatic breast cancer requires monitoring of the tumor burden to determine the response to treatment, and improved biomarkers are needed. Biomarkers such as cancer antigen 15-3 (CA 15-3) and circulating tumor cells have been widely studied. However, circulating cell-free DNA carrying tumor-specific alterations (circulating tumor DNA) has not been extensively investigated or compared with other circulating biomarkers in breast cancer. We compared the radiographic imaging of tumors with the assay of circulating tumor DNA, CA 15-3, and circulating tumor cells in 30 women with metastatic breast cancer who were receiving systemic therapy. We used targeted or whole-genome sequencing to identify somatic genomic alterations and designed personalized assays to quantify circulating tumor DNA in serially collected plasma specimens. CA 15-3 levels and numbers of circulating tumor cells were measured at identical time points. Circulating tumor DNA was successfully detected in 29 of the 30 women (97%) in whom somatic genomic alterations were identified; CA 15-3 and circulating tumor cells were detected in 21 of 27 women (78%) and 26 of 30 women (87%), respectively. Circulating tumor DNA levels showed a greater dynamic range, and greater correlation with changes in tumor burden, than did CA 15-3 or circulating tumor cells. Among the measures tested, circulating tumor DNA provided the earliest measure of treatment response in 10 of 19 women (53%). This proof-of-concept analysis showed that circulating tumor DNA is an informative, inherently specific, and highly sensitive biomarker of metastatic breast cancer. (Funded by Cancer Research UK and others.).


PPAR-γ agonists and their effects on IGF-I receptor signaling: Implications for cancer.
It is now well established that the development and progression of a variety of human malignancies are associated with dysregulated activity of the insulin-like growth factor (IGF) system. In this regard, promising drugs have been developed to target the IGF-I receptor or its ligands. These therapies are limited by the development of insulin resistance and compensatory hyperinsulinemia, which in turn, may stimulate cancer growth. Novel therapeutic approaches are, therefore, required. Synthetic PPAR-γ agonists, such as thiazolidinediones (TZDs), are drugs universally used as antidiabetic agents in patients with type 2 diabetes. In addition of acting as insulin sensitizers, PPAR-γ agonists mediate in vitro and in vivo pleiotropic anticancer effects. At least some of these effects appear to be linked with the downregulation of the IGF system, which is induced by the cross-talk of PPAR-γ agonists with multiple components of the IGF system signaling. As hyperinsulinemia is an emerging cancer risk factor, the insulin lowering action of PPAR-γ agonists may be expected to be also beneficial to reduce cancer development and/or progression. In light of these evidences, TZDs or other PPAR-γ agonists may be exploited in those tumors "addicted" to the IGF signaling and/or in tumors occurring in hyperinsulinemic patients.


The significance of walking from the perspective of people with Parkinson's disease.
Parkinson's disease (PD) is associated with progressive mobility and walking difficulties. Although these aspects have been found to be central from a patient perspective, the perceived significance of walking is less well understood. To explore the significance of walking as perceived and experienced by individuals with PD. Eleven persons with PD (seven men; median age, 71 years; median PD duration, 7 years) participated. Data were collected through semi-structured interviews, which were recorded and transcribed verbatim. Data were qualitatively analysed by systematic text condensation. The ability to walk had a complex and multifaceted impact on the participants, including physical, psychological and emotional aspects as well as on the ability to be active in daily life and to participate in society. The central role of coping strategies was prominent in filtering emotional reactions to physical changes, and when managing the activities and participation in everyday situations. The sense of unpredictability, uncertainty and loss of control were underlying phenomena in all categories. Furthermore, inability to manage walking difficulties had a negative impact on the participants' self-concept. The central meaning of being able to walk appears to be intimately linked to an individual's social identity, emotional well-being and integrity. Consequently, being able to walk independently was a prerequisite to an autonomous life and participation in society. This implies that rehabilitation and other mobility interventions also need to consider individual emotional, psychological, and social implications, and to facilitate appropriate compensatory and coping strategies.


Resimmune, an anti-CD3ε recombinant immunotoxin, induces durable remissions in patients with cutaneous T-cell lymphoma.
Resimmune is a second-generation recombinant immunotoxin composed of the catalytic and translocation domains of diphtheria toxin fused to two single chain antibody fragments reactive with the extracellular domain of CD3ε. We gave intravenous infusions of Resimmune 2.5 - 11.25 μg/kg over 15 minutes to 30 patients (25 with cutaneous T-cell lymphoma, 3 with peripheral T-cell lymphoma, 1 with T-cell large granular lymphocytic leukemia and 1 with T-cell prolymphocytic leukemia) in an inter-patient dose escalation trial. The most common adverse events were fever, chills, hypotension, edema, hypoalbuminemia, hypophosphatemia, and transaminasemia. Among the 25 patients with cutaneous T-cell lymphoma, there were nine responses for a response rate of 36% (95% CI, 18%-57%) including four complete remissions (16%, 95% CI, 5%-36%). The durations of the complete remissions were 72+, 72+, 60+ and 38+ months. There were five partial remissions lasting 3, 3, 3+, 6+ and 14 months. Of 17 patients with a modified skin weighted assessment tool score <50, 17 patients with stage IB/IIB, and 11 patients with both a score <50 and stage IB/IIB, nine (53%), eight (47%), and eight (73%) had responses, respectively. Further studies of Resimmune in patients with low tumor burden, stage IB-IIB cutaneous T-cell lymphoma are warranted. This trial is registered at clinicaltrials.gov as #NCT00611208.


Neuropsychological evidence that somatic stimuli are spatially coded according to multiple frames of reference in a stroke patient with tactile extinction.
A right-brain damaged patient with pure tactile extinction was asked to report series of single or double light, brief touches delivered to both hands, the thumb or the pinkie of a single hand, the sides of a single index. The stimulated hand was positioned palm up or palm down, in front of or behind the patient, in anatomic or crossed position. In double touch conditions, stimuli coded as left-sided were extinguished not only when delivered to both hands, but also when delivered on a single hand or a single finger. The findings suggest that tactile stimuli may be coded as left or right according to multiple body anchors that are dynamically scaled from the corporeal midline, to axes centered on a single hand or finger.


Direct plasma radioimmunoassay for rat amylin-(1-37): concentrations with acquired and genetic obesity.
Amylin (islet-associated polypeptide) is a 37-amino acid peptide that is cosecreted with insulin from the pancreatic beta-cell. Accurate measurement of its plasma levels is important for delineating the physiological range over which amylin acts. We describe a reproducible, highly specific, and sensitive radioimmunoassay for direct measurement of plasma amylin-(1-37). We measured changes in portal and systemic plasma amylin and insulin in three groups of anesthetized rats: lean young adult and old adult Wistar rats with acquired obesity, and Wistar fatty [WDF/TaFa (fa/fa)] rats, a model of genetic obesity and insulin resistance derived from the Wistar strain. Changes in response to fasting, feeding, and intravenous stimulation with glucose plus arginine were assessed. We find that the amylin-to-insulin ratio is constant in fasted or fed young and old rats because of proportionate increases in both entities with aging. In genetically obese Wistar rats, amylin and insulin levels are three- to tenfold higher than in lean young or obese old normal controls. Islet stimulation by feeding or intravenous glucose plus arginine resulted in a decreased amylin-to-insulin molar ratio in all groups. When normalized for the degree of islet stimulation, amylin-to-insulin ratios were significantly elevated in genetically obese vs. normal rats, both in the portal and systemic circulation. These results demonstrate that aging-related weight gain in normal rats is associated with moderate and proportional increases in amylin and insulin, whereas genetic obesity is characterized by elevated amylin and an increased amylin-to-insulin ratio. Implications for the pathogenesis of insulin resistance and obesity are discussed.


"Losing My Self": A Poet's Ironies and a Daughter's Reflections on Dementia.
What can we learn about the experience of dementia and about ways of being human when a poet describes her forgetting? My mother, the poet Shirley Kaufman, died in 2016 at the age of 93. She had dementia for many years. By her mid-80s, many of her poems probed the experience of forgetting, including questions about the nature of self, memory, and thought, and the location of the past and the future. Her experience of forgetting revealed in her ninth and last collection, the widely acclaimed Ezekiel's Wheels (2009), serves as my entry point for an exploration of the self as it becomes dislocated in time yet remains attuned to and consciously aware of that dislocation. That dual quality of my mother's experience is the central, ironic tension explored in this essay. An intimate ethnography, this essay connects my mother's late-life poetry with my own experience, both as a daughter who watched her mother's state of mind unfold and as an anthropologist with extensive experience studying aging, late-life identity, illness, and the culture of medicine. Through the insights of my mother's poetic language I show how dementia can expose ironic features of selfhood, communication, and life itself that are worth our discernment. My hope is that my analysis may focus our gaze on the inherent tension in this form of life and teach us something new about identity, memory, and what is shared among us, those with and without the condition we call dementia.


Generation of IgG aggregates by the myeloperoxidase-hydrogen peroxide system.
Incubation of native human 125I-IgG with polymorphonuclear neutrophil (PMN) peroxidase-containing granules or with purified myeloperoxidase (MPO) in the presence of H2O2 and a suitable hydrogen donor such as catechol generated large amounts of heavy IgG aggregates. Short-term incubation (15 to 60 min) of native 125I-IgG (400 microgram) with MPO-containing granules or with purified MPO (1.5 microgram) in the presence of H2O2 (0.036 to 0.36 mumol) and catechol (0.2 mumol) resulted in the generation of 8 to 100 microgram of heavy IgG aggregates (3 X 10(5) to 4 X 10(6) daltons). Aggregate formation was completely abolished by the omission of H2O2 or catechol, and by the addition of catalase, sodium azide, or cyanide. IgG aggregates were also generated with tyrosinase, tyrosine, and atmospheric oxygen. These results indicate that aggregation was due to MPO-H2O2-mediated oxidation of catechol to orthoquinone, which was deemed to be directly responsible for cross-linking by non-enzymic biochemical reactions. The IgG aggregates generated were shown to behave as typical immune complexes in that they consumed C, were detected by the solid-phase C1q and Raji cell assays, and were precipitable by monoclonal rheumatoid factor. This nonspecific oxidative protein-aggregation reaction may play an important role in the pathogenesis of tissue injury in acute and chronic inflammatory processes and in drug reactions. It could also provide an explanation for the frequent detection of circulating immune complex-like material in a large variety of acute and chronic inflammatory states.


Adaptations of Interferon Regulatory Factor 3 with Transition from Terrestrial to Aquatic Life.
Interferon regulatory factor 3 (IRF3) and IRF7 are closely related IRF members and the major factors for the induction of interferons, a key component in vertebrate innate immunity. However, there is limited knowledge regarding the evolution and adaptation of those IRFs to the environments. Two unique motifs in IRF3 and 7 were identified. One motif, GASSL, is highly conserved throughout the evolution of IRF3 and 7 and located in the signal response domain. Another motif, DPHK, is in the DNA-binding domain. The ancestral protein of IRF3 and 7 seemed to possess the DPHK motif. In the ray-finned fish lineage, while the DPHK is maintained in IRF7, the motif in IRF3 is changed to NPHK with a D → N amino acid substitution. The D → N substitution are also found in amphibian IRF3 but not in amphibian IRF7. Terrestrial animals such as reptiles and mammals predominantly use DPHK sequences in both IRF3 and 7. However, the D → N substitution in IRF3 DPHK is again found in cetaceans such as whales and dolphins as well as in marsupials. These observations suggest that the D → N substitutions in the IRF3 DPHK motif is likely to be associated with vertebrate's adaptations to aquatic environments and other environmental changes.


Prevalence and factors associated with low back pain among nurses at a regional hospital in KwaZulu-Natal, South Africa.
Low back pain (LBP) is a public health problem worldwide and is a common cause of work-related disorder among workers, especially in the nursing profession. Recruitment and retention of nurses is a challenge, and the nursing shortage has been exacerbated by the burden of occupational injuries such as LBP and related disabilities. The physiotherapy clinical records revealed that caseload of nurses presenting for the management of LBP was increasing. The prevalence and factors associated with LBP were unclear. A cross-sectional study design with an analytic component was implemented. Data were collected utilising a self-administered questionnaire to determine the prevalence and factors associated with LBP among nurses at a regional hospital. Bivariate analyses were performed to determine the factors associated with LBP. The point prevalence of current LBP in nurses was 59%. The highest prevalence was recorded among enrolled nurses (54%), respondents aged 30-39 (46%), overweight respondents (58%) and those working in obstetrics and gynaecology (49%). Bending (p = 0.002), prolonged position (p = 0.03) and transferring patients (p = 0.004) were strongly associated with LBP. Nurses with more than 20 years in the profession reported a high prevalence of LBP. The prevalence of LBP was higher among the participants who were on six-month rotations (76%) compared with those on yearly rotation (16%). A high proportion of nurses reported to have LBP. Occupational factors are strongly associated with LBP. Education programmes on prevention and workplace interventions are required in order to reduce occupational injuries.


Probiotic use in at-risk populations.
To inform health care providers about quality standards for manufacture of probiotic products being recommended for at-risk patient populations. Probiotics are used in a variety of clinical settings, sometimes in at-risk populations for therapeutic endpoints. Although probiotics might not be approved as drugs, they are sometimes used for the prevention or treatment of disease. In the United States, and many regions of the world, probiotic products are marketed as dietary supplements (not drugs) and are therefore subject to different manufacturing and quality control standards than approved drugs are. Health care providers need to be assured that probiotic products used in at-risk populations are safe for this use. Pharmacists should require certificates of analysis, which document quality standards, from manufacturers of products stocked in hospital formularies or other pharmacies dispensing to at-risk people. Although responsible manufacturers use stringent quality standards on their processes and finished products, using a third party to verify compliance with manufacturing and accuracy of product labeling adds assurance to end users that the product is of high quality. It is in patients' best interest to use probiotics in the prevention and treatment of conditions when the evidence is convincing. To protect high-risk patients, probiotic products should meet stringent microbiological standards. Product testing results should be available for review before recommending probiotic products to at-risk individuals. For products used in at-risk populations, manufacturers should provide this information or participate in a third-party verification program that certifies compliance.


Disruption of writing processes by the semanticity of background speech.
Previous studies have noted that writing processes are impaired by task-irrelevant background sound. However, what makes sound distracting to writing processes has remained unaddressed. The experiment reported here investigated whether the semanticity of irrelevant speech contributes to disruption of writing processes beyond the acoustic properties of the sound. The participants wrote stories against a background of normal speech, spectrally-rotated speech (i.e., a meaningless sound with marked acoustic resemblance to speech) or silence. Normal speech impaired quantitative (e.g., number of characters produced) and qualitative/semantic (e.g., uncorrected typing errors, proposition generation) aspects of the written material, in comparison with the other two sound conditions, and it increased the duration of pauses between words. No difference was found between the silent and the rotated-speech condition. These results suggest that writing is susceptible to disruption from the semanticity of speech but not especially susceptible to disruption from the acoustic properties of speech.


Chord-based versus voxel-based methods of electron transport in the skeletal tissues.
Anatomic models needed for internal dose assessment have traditionally been developed using mathematical surface equations to define organ boundaries, shapes, and their positions within the body. Many researchers, however, are now advocating the use of tomographic models created from segmented patient computed tomography (CT) or magnetic resonance (MR) scans. In the skeleton, however, the tissue structures of the bone trabeculae, marrow cavities, and endosteal layer are exceedingly small and of complex shape, and thus do not lend themselves easily to either stylistic representations or in-vivo CT imaging. Historically, the problem of modeling the skeletal tissues has been addressed through the development of chord-based methods of radiation particle transport, as given by studies at the University of Leeds (Leeds, U.K.) using a 44-year male subject. We have proposed an alternative approach to skeletal dosimetry in which excised sections of marrow-intact cadaver spongiosa are imaged directly via microCT scanning. The cadaver selected for initial investigation of this technique was a 66-year male subject of nominal body mass index (22.7 kg m(-2)). The objectives of the present study were to compare chord-based versus voxel-based methods of skeletal dosimetry using data from the UF 66-year male subject. Good agreement between chord-based and voxel-based transport was noted for marrow irradiation by either bone surface or bone volume sources up to 500-1000 keV (depending upon the skeletal site). In contrast, chord-based models of electron transport yielded consistently lower values of the self-absorbed fraction to marrow tissues than seen under voxel-based transport at energies above 100 keV, a feature directly attributed to the inability of chord-based models to account for nonlinear electron trajectories. Significant differences were also noted in the dosimetry of the endosteal layer (for all source tissues), with chord-based transport predicting a higher fraction of energy deposition than given by voxel-based transport (average factor of about 1.6). The study supports future use of voxel-based skeletal models which (1) permit nonlinear electron trajectories across the skeletal tissues, (2) do not rely on mathematical algorithms for treating the endosteal tissue layer, and (3) do not implicitly assume independence of marrow and bone trajectories as is the case for chord-based skeletal models.


[Relationship between lead levels in colostrum, dietary intake, and socioeconomic characteristics of puerperal women in Goiânia, Brazil].
To estimate the levels of lead in human colostrum and investigate whether they are correlated with dietary intake and socioeconomic factors in puerperal women. This cross-sectional study was carried out with 80 puerperal women from a maternity hospital in Goiânia, Brazil. Data were collected between July and December 2008. A questionnaire was used to collect socioeconomic and lifestyle information. A dietary intake frequency questionnaire was also applied. After that, colostrum samples were collected and analyzed using flame atomic absorption spectrometry to quantify lead levels. The correlation between lead levels, socioeconomic variables, and dietary intake was calculated. The mean concentration of lead in colostrum samples was 6.88 μg/L, and the median concentration was 4.65 μg/L. There was no association between socioeconomic aspects and lead levels. The intake of saturated fatty acids, eggs, pork meats, chicken, and chicken products (r = -0.26 and P = 0.020) was inversely correlated with the levels of lead in human colostrum, whereas the intake of squash (r = 0.26; P = 0.019) and eggplant (r = 0.27; P = 0.015) was positively correlated with the levels of lead. The correlation observed for cooked squash, eggs, pork and chicken meats, and chicken products was maintained after logistic regression adjustment. The presence of lead was detected in human colostrum in this sample, probably as a result of dietary exposure. In addition to a correlation between presence of lead and dietary intake, the present findings suggest that specific nutrients may result in increased or decreased levels of lead, without a direct relationship with socioeconomic factors.


Decreased tryptophan and increased kynurenine levels in mastocytosis associated with digestive symptoms.
The main metabolism pathway of tryptophan is protein formation, but it can also be metabolized into serotonin and kynurenine. Indoleamine 2,3-dioxygenase (IDO) is the enzyme that catalyzes the degradation of tryptophan into kynurenine. Mastocytosis is a heterogeneous disease characterized by mast cell accumulation in various tissues with 57% of patients having gastrointestinal involvement. We studied tryptophan metabolism in mastocytosis patients displaying or not gastrointestinal features and healthy subjects (n = 26 in each group). Mastocytosis patients with digestive symptoms displayed significantly increased kynurenine level and IDO activity as compared to healthy controls and mastocytosis patients without digestive symptoms. This could be linked to mast cell-mediated digestive inflammation among patients with mastocytosis. This work is the first focusing on kynurenine pathway in a mast cell disease and could help to understand the pathogenesis of digestive features in mastocytosis as well as in other mast cell-mediated diseases.


Rapamycin inhibits transforming growth factor beta 1 induced myofibroblast differentiation via the phosphorylated-phosphatidylinositol 3-kinase mammalian target of rapamycin signal pathways in nasal polyp-derived fibroblasts.
Rapamycin has antiproliferative and antifibrogenic effects in vitro and in vivo. The purpose of this study was to evaluate the effects of rapamycin on transforming growth factor (TGF) beta 1 induced myofibroblast differentiation (alpha smooth-muscle actin [SMA]), extracellular matrix production, and collagen contraction in nasal polyp-derived fibroblasts (NPDF). The underlying molecular mechanisms of rapamycin were also determined in NPDFs. NPDFs were grown in culture and transformed into myofibroblasts by using TGF beta 1 (5 ng/mL). For cytotoxicity evaluation, a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay was used. Expression levels of alpha SMA, phosphorylated phosphatidylinositol 3-kinase (PI3K), and phosphorylated mammalian target of rapamycin (mTOR) were determined by using Western blot, reverse transcription-polymerase chain reaction, and immunofluorescence staining. The total amount of collagen was analyzed by using the Sircol collagen assay, and contractile activity was measured with a collagen gel contraction assay. Silencing mTOR with mTOR-specific small interference RNA was determined by using reverse transcription-polymerase chain reaction. Whereas rapamycin (range, 0-400 nM) had no significant cytotoxic effects on TGF beta 1 induced NPDFs, it significantly reduced the expression levels of alpha-SMA in TGF beta 1 induced NPDFs in a dose-dependent manner. TGF beta 1 induced collagen production and collagen contraction were significantly inhibited by rapamycin treatment. Rapamycin also attenuated the TGF beta 1 induced activation of PI3K and mTOR, and its inhibitory effects were similar to those of mTOR silencing and a specific PI3K inhibitor. Rapamycin inhibited TGF beta 1 induced myofibroblast differentiation, extracellular matrix production, and collagen contraction through the PI3K/mTOR signal pathways in NPDFs.


Dementia risk in renal dysfunction: A systematic review and meta-analysis of prospective studies.
Renal dysfunction has been linked with increased risk for cognitive impairment and dementia, but studies are conflicting. For that reason, the aim of the present systematic review and meta-analysis is to summarize the best available evidence on the prospective association between potential markers of renal dysfunction and development of cognitive impairment or dementia. Medline, Embase, and Cochrane Database of Systematic Reviews were searched for potential publications until August 1, 2016. Studies were eligible if they fulfilled the following criteria: population-based study, prospective design, ≥100 participants, aged ≥45 years, ≥1 year follow-up, and cognition/dementia outcomes. Where appropriate, random effects meta-analyses were conducted yielding pooled odds ratios (OR) and 95% confidence intervals (CI). Twenty-two out of 8,494 abstracts fulfilled the eligibility criteria. Sufficient evidence was found for albuminuria, mixed results for estimated glomerular filtration rate (eGFR), insufficient support for cystatin C, and tentative evidence for serum creatinine and creatinine clearance. Meta-analyses of 5 studies representing 27,805 persons showed a 35% increased risk of cognitive impairment or dementia in those with albuminuria (OR 1.35, 95% CI 1.06-1.73, p = 0.015), whereas eGFR <60 mL/min/1.73 m2 showed no significant association (OR 1.28, 95% CI 0.99-1.65, p = 0.063). No meta-analyses could be done for serum creatinine, creatinine clearance, or cystatin C. The overall evidence for an association between renal dysfunction and cognitive impairment or dementia is modest. Evidence suggests that albuminuria is associated with higher odds of developing cognitive impairment or dementia.


Computed tomography attenuation values of ascites are helpful to predict perforation site.
To evaluate the effect of computed tomography (CT) attenuation values of ascites on gastrointestinal (GI) perforation site prediction. The CT attenuation values of the ascites from 51 patients with GI perforations were measured by volume rendering to calculate the mean values. The effect of the CT attenuation values of the ascites on perforation site prediction and postoperative complications was evaluated. Of 24 patients with colorectal perforations, the CT attenuation values of ascites were significantly higher than those in patients with perforations at other sites [22.5 Hounsfield units (HU) vs 16.5 HU, respectively, P = 0.006]. Colorectal perforation was significantly associated with postoperative complications (P = 0.038). The prediction rate of colorectal perforation using attenuation values as an auxiliary diagnosis improved by 9.8% compared to that of CT findings alone (92.2% vs 82.4%). The CT attenuation values of ascites could facilitate the prediction of perforation sites and postoperative complications in GI perforations, particularly in cases in which the perforation sites are difficult to predict by CT findings alone.


Surviving Sepsis Campaign: guidelines on the management of critically ill adults with Coronavirus Disease 2019 (COVID-19).
The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of a rapidly spreading illness, Coronavirus Disease 2019 (COVID-19), affecting thousands of people around the world. Urgent guidance for clinicians caring for the sickest of these patients is needed. We formed a panel of 36 experts from 12 countries. All panel members completed the World Health Organization conflict of interest disclosure form. The panel proposed 53 questions that are relevant to the management of COVID-19 in the ICU. We searched the literature for direct and indirect evidence on the management of COVID-19 in critically ill patients in the ICU. We identified relevant and recent systematic reviews on most questions relating to supportive care. We assessed the certainty in the evidence using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) approach, then generated recommendations based on the balance between benefit and harm, resource and cost implications, equity, and feasibility. Recommendations were either strong or weak, or in the form of best practice recommendations. The Surviving Sepsis Campaign COVID-19 panel issued 54 statements, of which 4 are best practice statements, 9 are strong recommendations, and 35 are weak recommendations. No recommendation was provided for 6 questions. The topics were: (1) infection control, (2) laboratory diagnosis and specimens, (3) hemodynamic support, (4) ventilatory support, and (5) COVID-19 therapy. The Surviving Sepsis Campaign COVID-19 panel issued several recommendations to help support healthcare workers caring for critically ill ICU patients with COVID-19. When available, we will provide new recommendations in further releases of these guidelines.


Proximal extension of backwash ileitis in ulcerative-colitis - associated colon cancer.
Extension of the inflammatory process into the terminal ileum in ulcerative colitis is termed backwash ileitis. Ulcerative colitis patients with pancolitis and backwash ileitis may be at increased risk of colorectal carcinoma. A case of ulcerative colitis-associated cancer with backwash ileitis, which extended proximally over time, is described. A 67-year-old man was diagnosed with pancolitis; despite prednisolone and mesalazine treatment, he showed no improvement. Although initial endoscopic examination showed no abnormalities in the terminal ileum, a repeat colonoscopy 4 months later revealed erythema, absence of the vascular pattern, mucosal friability, and erosions within 30 cm of the terminal ileum in a continuous fashion from the cecum, and a flat, elevated lesion was found in the transverse colon. On histopathologic examination, the patient had nonspecific, active, chronic inflammation of the ileal mucosa, consistent with backwash ileitis, and colonic adenocarcinoma. Despite additional treatment with leukocytapheresis, the backwash ileitis progressed, with increased severity of mucosal inflammation and extensive erosions, 20 cm proximal to the primarily affected site of the ileum on repeat colonoscopy 2 months later. The patient underwent a proctocolectomy and excision of the terminal ileum with an ileostomy. Histopathology of the surgical specimen revealed marked backwash ileitis and submucosal, well-differentiated adenocarcinoma. The patient has remained well for 5 years after surgery.


A SNAP coupled microresonator delay line.
A delay line fabricated of a chain of SNAP (Surface Nanoscale Axial Photonics) coupled microresonators is demonstrated. In contrast to resonant delay lines demonstrated to date, the slow light in this structure is enhanced by the 2R (Rotation + Reflection) effect realized due to the 3D propagation of light along the surface of a SNAP fiber. Here, the delay line coupled to a single input/output waveguide (i.e., operating in the reflection mode) is considered. Depending on the coupling parameters and loss, the delay time in this device is either proportional to the density of resonances averaged over the pulse spectrum or tends to zero. The delay line is fabricated of 20 coupled microresonators with the total length of 1.2 mm and footprint area of 0.05 mm2. It exhibits the record low insertion loss (< 3 dB), small speed of light (<c/250), and large (>1 ns) delay time along the 0.1 nm bandwidth achieved for the miniature microresonator delay lines. The feasibility of significant improvement of the SNAP delay line characteristics (larger delay time and bandwidth, smaller losses and dimensions, and anti-reflecting apodization) is discussed.


Proteogenomic analysis of Mycobacterium tuberculosis by high resolution mass spectrometry.
The genome sequencing of H37Rv strain of Mycobacterium tuberculosis was completed in 1998 followed by the whole genome sequencing of a clinical isolate, CDC1551 in 2002. Since then, the genomic sequences of a number of other strains have become available making it one of the better studied pathogenic bacterial species at the genomic level. However, annotation of its genome remains challenging because of high GC content and dissimilarity to other model prokaryotes. To this end, we carried out an in-depth proteogenomic analysis of the M. tuberculosis H37Rv strain using Fourier transform mass spectrometry with high resolution at both MS and tandem MS levels. In all, we identified 3176 proteins from Mycobacterium tuberculosis representing ~80% of its total predicted gene count. In addition to protein database search, we carried out a genome database search, which led to identification of ~250 novel peptides. Based on these novel genome search-specific peptides, we discovered 41 novel protein coding genes in the H37Rv genome. Using peptide evidence and alternative gene prediction tools, we also corrected 79 gene models. Finally, mass spectrometric data from N terminus-derived peptides confirmed 727 existing annotations for translational start sites while correcting those for 33 proteins. We report creation of a high confidence set of protein coding regions in Mycobacterium tuberculosis genome obtained by high resolution tandem mass-spectrometry at both precursor and fragment detection steps for the first time. This proteogenomic approach should be generally applicable to other organisms whose genomes have already been sequenced for obtaining a more accurate catalogue of protein-coding genes.


Evaluation of unintended effects in the composition of tomatoes expressing a human immunoglobulin A against rotavirus.
The production of neutralizing immunoglobulin A (IgA) in edible fruits as a means of oral passive immunization is a promising strategy for the inexpensive treatment of mucosal diseases. This approach is based on the assumption that the edible status remains unaltered in the immunoglobulin-expressing fruit, and therefore extensive purification is not required for mucosal delivery. However, unintended effects associated with IgA expression such as toxic secondary metabolites and protein allergens cannot be dismissed a priori and need to be investigated. This paper describes a collection of independent transgenic tomato lines expressing a neutralizing human IgA against rotavirus, a mucosal pathogen producing severe diarrhea episodes. This collection was used to evaluate possible unintended effects associated with recombinant IgA expression. A comparative analysis of protein and secondary metabolite profiles using wild type lines and other commercial varieties failed to find unsafe features significantly associated with IgA expression. Preliminary, the data indicate that formulations derived from IgA tomatoes are as safe for consumption as equivalent formulations derived from wild type tomatoes.


The effect of chronic cytomegalovirus infection on pneumococcal vaccine responses.
Immune function declines with age and has been associated with reduced vaccine responsiveness. Chronic infection with cytomegalovirus (CMV) has been proposed as a contributor to poorer responses in older adults. A pneumococcal vaccine has been recommended in the United Kingdom for those aged >65 years since 2003 to prevent pneumococcal disease. We evaluated the effect of age and CMV status on pneumococcal vaccine responses in 348 individuals aged 50-70 years. We found participant age to be associated with serotype-specific and functional antibody titers after pneumococcal vaccination, with a mean 6.2% (95% confidence interval, 2.9%-9.5%) reduction in postvaccination functional antibody titers per year. CMV status was not associated with serotype-specific immunoglobulin G concentrations or functional antibody titers after pneumococcal vaccination. However, CMV seropositivity was associated with higher levels of prevaccination functional antibody for 4 of 7 pneumococcal serotypes assessed. These data imply that CMV infection is not directly responsible for the decline in pneumococcal vaccine responses seen with age but suggest that CMV-seropositive individuals differ in their natural exposure to pneumococci or have altered mucosal immune responses after colonization with this organism.


Pleistocene climates in the atlantic and pacific oceans: a reevaluated comparison based on deep-sea sediments.
Variations of the Globorotalia menardii complex in cores from the Indian Ocean can be interpreted as indicating climatic changes that are opposite to trends exhibited by the total planktonic fauna. The questionable value of correlations between different water masses based on a single species can be shown by Neogloboquadrina dutertrei subcretacea in cores obtained off the coast of California and Baja California. This information, in addition to previous correlations between Quaternary cores of the Atlantic and Pacific oceans, indicates that the Pleistocene history of the two oceans was parallel.


All-fiber bidirectional optical modulator derives from the microfiber coated with ITO electrode.
A cheap, compact, and simply prepared all-fiber bidirectional optical modulator based on the Pockels effect of water and the band population effect was first, to the best of our knowledge, proposed and demonstrated. The transparent conductive oxide indium-tin-oxide (ITO) was coated on the surface of a nonadiabatic microfiber and first used as a modulating electrode on the microfiber. The device was realized by just submerging the microfiber in water. With supplying an electric field perpendicular to the interface between the microfiber and water, the refractive index was modulated in the electric double layer near the tapered region of the microfiber, under the Pockels effect of water. Subsequently, the interference spectrum was modulated. Meanwhile, the intensity of the light was modulated due to the band population effect in the space-charge layer. In this Letter, the proposed all-fiber optical modulator can realize simultaneous bidirectional modulation of the phase and intensity of output light. Experimentally, the maximum phase shift and the extinction ratio were 4.38 nm and 4.87 dB at 1550 nm, respectively. Significantly, the work used the Pockels effect of water and the band population effect to realize an all-fiber optical modulator, showing great potential in the optical phase modulators, optical switches, and electric field sensors.


Sustained increase in α5GABAA receptor function impairs memory after anesthesia.
Many patients who undergo general anesthesia and surgery experience cognitive dysfunction, particularly memory deficits that can persist for days to months. The mechanisms underlying this postoperative cognitive dysfunction in the adult brain remain poorly understood. Depression of brain function during anesthesia is attributed primarily to increased activity of γ-aminobutyric acid type A receptors (GABA(A)Rs), and it is assumed that once the anesthetic drug is eliminated, the activity of GABA(A)Rs rapidly returns to baseline and these receptors no longer impair memory. Here, using a murine model, we found that a single in vivo treatment with the injectable anesthetic etomidate increased a tonic inhibitory current generated by α5 subunit-containing GABA(A)Rs (α5GABA(A)Rs) and cell-surface expression of α5GABA(A)Rs for at least 1 week. The sustained increase in α5GABA(A)R activity impaired memory performance and synaptic plasticity in the hippocampus. Inhibition of α5GABA(A)Rs completely reversed the memory deficits after anesthesia. Similarly, the inhaled anesthetic isoflurane triggered a persistent increase in tonic current and cell-surface expression of α5GABA(A)Rs. Thus, α5GABA(A)R function does not return to baseline after the anesthetic is eliminated, suggesting a mechanism to account for persistent memory deficits after general anesthesia.


Relationships of brain white matter microstructure with clinical and MR measures in relapsing-remitting multiple sclerosis.
To assess the relationships of microstructural damage in the cerebral white matter (WM), as measured by diffusion tensor imaging (DTI), with clinical parameters and magnetic resonance imaging (MRI) measures of focal tissue damage in patients with multiple sclerosis (MS). Forty-five relapsing-remitting (RR) MS patients (12 male, 33 female; median age = 29 years, Expanded Disability Status Scale (EDSS) = 1.5, disease duration = 3 years) were studied. T2-lesion masks were created and voxelwise DTI analyses performed with Tract-Based Spatial Statistics (TBSS). T2-lesion volume (T2-LV) was significantly (P < 0.05, corrected) correlated with fractional anisotropy (FA) in both lesions and normal-appearing WM (NAWM). Relationships (P = 0.08, corrected) between increasing EDSS score and decreasing FA were found in the splenium of the corpus callosum (sCC) and along the pyramidal tract (PY). All FA associations were driven by changes in the perpendicular (to primary tract direction) diffusivity. No significant global and voxelwise FA changes were found over a 2-year follow-up. FA changes related to clinical disability in RR-MS patients with minor clinical disability are localized to specific WM tracts such as the sCC and PY and are driven by changes in perpendicular diffusivity both within lesions and NAWM. Longitudinal DTI measurements do not seem able to chart the early disease course in the WM of MS patients.


Innate responsiveness of CD8 memory T-cell populations nonspecifically inhibits allergic sensitization.
Infection or stimulation of the innate immune system by nonspecific microbial antigens is thought to educate the immune system to respond appropriately to allergens, preventing allergy. To determine the immunologic pathways that might explain how infection/microbial exposure inhibits allergic sensitization. Immunologic studies of non-antigen-specific functions of CD8 memory cells, their maturation in vivo, and their effects in a mouse asthma model, to test the hypothesis that CD8 memory is shaped by innate immunity in a way that can inhibit allergic disease. We found that CD8 memory T-cell (CD8 Tm) populations bridge innate and adaptive immunity by responding to either antigen or cytokines alone. CD8 Tm populations partially subvert the clonal selection process by activating their neighbors through induction of dendritic cell IL-12. Stimulation of innate or acquired immunity in the lung or gut causes expansion/maturation of CD8 Tm populations, which provide an early source of cytokines, enhance T(H)1 immunity, and inhibit allergic sensitization and airway inflammation/hyperresponsiveness in a non-antigen-specific fashion. CD8 T-cell-mediated immune memory is long-lived and can retain its capacity for rapid cytokine release in a nonantigen-specific fashion. This novel type of memory enhances T(H)1 over T(H)2 immunity and prevents allergic sensitization after exposure to environmental antigens or infection.


Wound healing potential of antibacterial microneedles loaded with green tea extracts.
This study evaluates the utility of an antibacterial microneedle composed of green tea (GT) extract and hyaluronic acid (HA), for the efficient delivery of GT. These microneedles have the potential to be a patient-friendly method for the conventional sustained release of drugs. In this study, a fabrication method using a mold-based technique to produce GT/HA microneedles with a maximum area of ~50mm(2) with antibacterial properties was used to manufacture transdermal drug delivery systems. Fourier transform infrared (FTIR) spectrometry was carried out to observe the potential modifications in the microneedles, when incorporated with GT. The degradation rate of GT in GT/HA microneedles was controlled simply by adjusting the HA composition. The effects of different ratios of GT in the HA microneedles were determined by measuring the release properties. In HA microneedles loaded with 70% GT (GT70), a continuous higher release rate was sustained for 72h. The in vitro cytotoxicity assays demonstrated that GT/HA microneedles were not generally cytotoxic to Chinese hamster ovary cells (CHO-K1), human embryonic kidney cells (293T), and mouse muscle cells (C2C12), which were treated for 12 and 24h. Antimicrobial activity of the GT/HA microneedles was demonstrated by ~95% growth reduction of gram negative [Escherichia coli (E. coli), Pseudomonas putida (P. putida), and Salmonella typhimurium (S. typhimurium)] and gram positive bacteria [Staphylococcus aureus (S. Aureus) and Bacillus subtilis (B. subtilis)], with GT70. Furthermore, GT/HA microneedles reduced bacterial growth of infected wound sites in the skin and improved wound healing process of skin in rat model.


Yoga-based pulmonary rehabilitation for the management of dyspnea in coal miners with chronic obstructive pulmonary disease: A randomized controlled trial.
Coal mine dust exposure causes chronic airflow limitation in coal miners resulting in dyspnea, fatigue, and eventually chronic obstructive pulmonary disease (COPD). Yoga can alleviate dyspnea in COPD by improving ventilatory mechanics, reducing central neural drive, and partially restoring neuromechanical coupling of the respiratory system. To evaluate the effectiveness of Integrated Approach of Yoga Therapy (IAYT) in the management of dyspnea and fatigue in coal miners with COPD. Randomized, waitlist controlled, single-blind clinical trial. Eighty-one coal miners (36-60 years) with stable Stages II and III COPD were recruited. The yoga group received an IAYT module for COPD that included asanas, loosening exercises, breathing practices, pranayama, cyclic meditation, yogic counseling and lectures 90 min/day, 6 days/week for 12 weeks. Measurements of dyspnea and fatigue on the Borg scale, exercise capacity by the 6 min walk test, peripheral capillary oxygen saturation (SpO2%), and pulse rate (PR) using pulse oximetry were made before and after the intervention. Statistically significant within group reductions in dyspnea (P < 0.001), fatigue (P < 0.001) scores, PR (P < 0.001), and significant improvements in SpO2% (P < 0.001) and 6 min walk distance (P < 0.001) were observed in the yoga group; all except the last were significant compared to controls (P < 0.001). Findings indicate that IAYT benefits coal miners with COPD, reducing dyspnea; fatigue and PR, and improving functional performance and peripheral capillary SpO2%. Yoga can now be included as an adjunct to conventional therapy for pulmonary rehabilitation programs for COPD patients.


Fluorine-induced apoptosis and lipid peroxidation in human hair follicles in vitro.
Fluoride is an essential trace element for human body; however, exposure to high amounts of fluoride has been documented to be correlated with an increasing risk of hair loss. To date, little is known about the mechanism(s) of how fluoride affects hair follicles. Here, we demonstrated that middle (1.0 mmol/L) and high (10.0 mmol/L) concentrations of sodium fluoride (NaF) significantly inhibited hair follicle elongation in vitro, but low NaF (0.1 mmol/L) showed little influence. Moreover, treatment with high levels of NaF resulted in a marked increase in terminal dUTP nick end labeling-positive cells in the outer layer of the outer root sheath, the dermal sheath, and the lower bulb matrix surrounding dermal papilla. Furthermore, the enhanced apoptosis was coupled with an increased oxidative stress manifested as higher malondialdehyde content. Additionally, the presence of selenium considerably antagonized the effects of middle NaF on hair follicles, with regard to either the suppression of hair growth or the induction of oxidative stress and apoptosis. In conclusion, exposure to high levels of fluoride compromises hair follicle growth and accelerate cell apoptosis in vitro. The toxicity of fluoride can be reduced by selenium, at least partially via the suppression of intracellular oxidative stress.


A polymorphism in type I deiodinase is associated with circulating free insulin-like growth factor I levels and body composition in humans.
The interaction between the GH-IGF-I axis and thyroid hormone metabolism is complex and not fully understood. T(4) stimulates IGF-I activity in animals in the absence of GH. On the other hand, GH replacement therapy results in an increase in serum T(3) and a decrease in T(4) and rT(3) levels, suggesting a stimulation of type I deiodinase (D1) activity. Recently, we demonstrated the association of two polymorphisms in D1 (D1a-C/T; T = 34%, and D1b-A/G; G = 10%) with serum iodothyronine levels. Haplotype alleles were constructed, suggesting a lower activity of the D1 haplotype 2 allele (aT-bA) and a higher activity of the haplotype allele 3 (aC-bG). In this study, we investigated whether genetic variations in D1 are associated with the IGF-I system. In 156 blood donors and 350 elderly men, the association of the D1 haplotype alleles with circulating IGF-I and free IGF-I levels was studied. In addition, potential associations with muscle strength and body composition were investigated in the elderly population. Finally, the relation between serum iodothyronine levels and IGF-I levels was studied. In blood donors, haplotype allele 2 was associated with higher levels of free IGF-I (302.9 +/- 22.9 vs. 376.3 +/- 19.1 pg/ml, P = 0.02). In elderly men, haplotype allele 2 also showed an allele dose increase in free IGF-I levels (P(trend) = 0.01) and an allele dose decrease in serum T(3) levels (P(trend) = 0.01), independent of age. Carriers of the D1a-T variant also had a higher isometric grip strength (P = 0.047) and maximum leg extensor strength (P = 0.07) as well as a higher lean body mass (P = 0.03). In blood donors, T(4) and free T(4) were negatively correlated with total IGF-I levels (R = -0.18, P = 0.03 and R = -0.24, P = 0.003), whereas T(3) to T(4) and T(3) to reverse T(3) ratios were positively correlated with total IGF-I (R = 0.31, P < 0.001 and R = 0.18, P = 0.03). Free IGF-I showed a negative correlation with T(4) (R = -0.26, P = 0.001) and T(4)-binding globulin (R = -0.31, P < 0.001) and a positive correlation with T(3) to T(4) ratio (R = 0.21, P = 0.01). In conclusion, a polymorphism that results in a decreased D1 activity is associated with an increase in free IGF-I levels. The pathophysiological significance of this association with IGF-I is supported by an increased muscle strength and muscle mass in carriers of the D1 haplotype 2 allele in a population of elderly men. The association of D1 haplotype allele 2 with serum T(3) levels in the elderly population suggests a relative increase in its contribution to circulating T(3) in old age.


Clinical Profile and Treatment of COVID-19 Patients: Experiences from an Ethiopian Treatment Center.
COVID-19 is not well studied in Africa. Understanding the clinical profile and management of COVID-19 will help to plan better prevention and treatment strategies taking the local context into consideration. In this study, we described the clinical profile, treatment used, and outcomes of COVID-19 patients in one of the COVID-19 treatment centers of Ethiopia, Boru Meda Hospital. An institution-based retrospective cross-sectional study was carried out using medical records of COVID-19 patients who were admitted to Boru Meda Hospital with a positive reverse transcription (RT)-PCR result from May 9, 2020 to September 20, 2020. All patients with a positive RT-PCR were admitted to the hospital, regardless of symptom and severity status. A total of 279 COVID-19 patients were included in the final analysis. The median age of patients was 28 years (interquartile range 23-40). The majority (69.5%) were male. Around a quarter (n = 73; 26.2%) of the patients were symptomatic, of which cough (n = 49; 67.1%) and fever (n = 32; 43.8%) were common symptoms. Among symptomatic patients, 48 (65.8%) were mild, four (5.5%) moderate, 12 (16.4%) severe, and nine (12.3%) were critical. The case fatality rate was 2.1%. Hypertension, age older than 25 years, and HIV/AIDS were significantly associated with symptomatic infection. In this study, most of the COVID-19 patients were asymptomatic. However, the proportion of severe and critical patients among those with symptoms was high. More studies are needed to assess the effect of HIV/AIDS on the severity and mortality of COVID-19.


Homopolymer mutational hot spots mediate herpes simplex virus resistance to acyclovir.
In the majority of cases, the mechanism underlying the resistance to acyclovir (ACV) of herpes simplex viruses (HSVs) is thymidine kinase (TK) deficiency. Plaque isolates from eight ACV-resistant (ACVr) clinical isolates from AIDS patients, of which five reactivated, were sequenced to determine the genetic lesion within the tk gene conferring resistance and whether this may have correlated with reactivation potential. Mutations were clustered within two homopolymer nucleotide stretches. Three plaque isolates (1737-14, 90-150-3, and 89-650-5) had insertion mutations within a stretch of 7 guanosines, while two isolates (89-063-1 and 89-353-1) had frameshift mutations within a stretch of 6 cytosines (a deletion and an insertion, respectively). Mutations resulted in premature termination codons, and the predicted 28- and 32-kDa truncated TK products were detected by Western blot analysis of virus-infected cell extracts. The repair of one homopolymer frameshift mutation (in isolate 1737-14) restored TK activity, demonstrating that this mutation is the basis of TK deficiency. Of the five reactivated isolates, four were TK deficient and contained frameshift mutations while the fifth retained TK activity because of its altered-TK or Pol- phenotype. These data demonstrate that the majority of ACVr clinical isolates contain frameshift mutations within two long homopolymer nucleotide stretches which function as hot spots within the HSV tk gene and produce nonfunctional, truncated TK proteins.


Evolutionary genomics of the cold-adapted diatom Fragilariopsis cylindrus.
The Southern Ocean houses a diverse and productive community of organisms. Unicellular eukaryotic diatoms are the main primary producers in this environment, where photosynthesis is limited by low concentrations of dissolved iron and large seasonal fluctuations in light, temperature and the extent of sea ice. How diatoms have adapted to this extreme environment is largely unknown. Here we present insights into the genome evolution of a cold-adapted diatom from the Southern Ocean, Fragilariopsis cylindrus, based on a comparison with temperate diatoms. We find that approximately 24.7 per cent of the diploid F. cylindrus genome consists of genetic loci with alleles that are highly divergent (15.1 megabases of the total genome size of 61.1 megabases). These divergent alleles were differentially expressed across environmental conditions, including darkness, low iron, freezing, elevated temperature and increased CO2. Alleles with the largest ratio of non-synonymous to synonymous nucleotide substitutions also show the most pronounced condition-dependent expression, suggesting a correlation between diversifying selection and allelic differentiation. Divergent alleles may be involved in adaptation to environmental fluctuations in the Southern Ocean.


Tyrosine kinase inhibitor therapy and metabolic remodelling in papillary thyroid cancer.
Targeted therapy is increasingly used to manage metastatic papillary thyroid cancer. The focus of the present study was to examine glucose metabolism and tumor responses for thyroid cancer xenografts expressing the glycolytic pathway modulators platelet-derived growth factor receptor (PDGFR) and BRAFV600E. Radiolabelled glucose derivative [18F]FDG was used to analyze the effects of PDGFR blockade with imatinib, BRAF blockade with vemurafenib, as well as combined PDGFR and BRAF blockade in vitro and in vivo with PET. Dynamic PET data was correlated with immunohistochemistry staining and kinetic analysis for facilitative glucose transporter 1 (GLUT1) and hexokinase-II (HK2). Vemurafenib decreased [18F]FDG uptake in BCPAP cells in vitro; however, it was increased by ~70% with imatinib application to BCPAP cells. This metabolic response to tyrosine kinase inhibition required BRAFV600E as it was not seen in cell lines lacking mutated BRAF (TPC1). In xenografts, imatinib therapy in BCPAP thyroid tumour-bearing mice significantly increased [18F]FDG uptake and retention (>30%) in BCPAP tumours with PDGFRβ or both (α+β) isoforms. Kinetic analysis revealed that the increased glucose uptake is a consequence of increased phosphorylation and intracellular trapping of [18F]FDG confirmed by an increase in HK2 protein expression and activity, but not GLUT1 activity. BRAF inhibition alone, or combined PDGFR and BRAF inhibition, reduced (~60%) [18F]FDG uptake in both types of BCPAP (β or α+β) tumours. In terms of tumour growth, combination therapy with imatinib and vemurafenib led to a near abolition of the tumors (~90% reduction), but single therapy for BCPAP with PDGFRα expression was much less effective. In summary, imatinib led to a paradoxical increase of [18F]FDG uptake in xenografts that was reversed through BRAFV600E inhibition. The present data show that metabolic reprogramming in thyroid cancer occurs as a consequence of BRAF-mediated upregulation of HK2 expression that may permit tumour growth with isolated blockade of upstream tyrosine kinase receptors.


Development of two-dimensional qualitative visualization method for isoflavones secreted from soybean roots using sheets with immobilized bovine serum albumin.
A visualization method for the qualitative evaluation of soybean isoflavones secreted from soybean roots by transferring them onto a sheet with immobilized bovine serum albumin (BSA) was developed. BSA was chemically bonded onto a glass microfiber filter. The fluorescence quenching resulting from the interaction of BSA with soybean isoflavones such as daidzein and daidzin was utilized. Fluorescence images before and after soybean roots were placed in contact with the sheets with immobilized BSA were taken with an electron-multiplying charge-coupled device camera. The fluorescence quenching in the images was visualized and analyzed. Soybean isoflavones were extracted from the sheets for quantitative analysis, and the correlation coefficient between the quenched fluorescence intensity per sheet and the total amount of soybean isoflavones was 0.78 (p < 0.01), indicating a high correlation. The quenched fluorescence intensity was lower in pumpkin roots, which do not secrete soybean isoflavone. It was found from analyzed images that soybean isoflavone is secreted in larger amounts from the basal region of the taproot and the tips of the lateral roots of soybean.


Hemoglobin redux: combining neutron and X-ray diffraction with mass spectrometry to analyse the quaternary state of oxidized hemoglobins.
Improvements in neutron diffraction instrumentation are affording the opportunity to re-examine the structures of vertebrate hemoglobins and to interrogate proton and solvent position changes between the different quaternary states of the protein. For hemoglobins of unknown primary sequence, structural studies of cyanomethemoglobin (CNmetHb) are being used to help to resolve sequence ambiguity in the mass spectra. These studies have also provided additional structural evidence for the involvement of oxidized hemoglobin in the process of erythrocyte senescence. X-ray crystal studies of Tibetan snow leopard CNmetHb have shown that this protein crystallizes in the B state, a structure with a more open dyad, which possibly has relevance to RBC band 3 protein binding and erythrocyte senescence. R-state equine CNmetHb crystal studies elaborate the solvent differences in the switch and hinge region compared with a human deoxyhemoglobin T-state neutron structure. Lastly, comparison of histidine protonation between the T and R state should enumerate the Bohr-effect protons.


The influence of micronutrients on cognitive function and performance.
There has been much media speculation (often sensationalist and conflicting) regarding the potential influence of micronutrients on cognitive function and performance. Our aim was to identify the micronutrients specifically implicated in cognitive function and to review the literature to identify original sources underlying the media coverage. Literature searches were carried out to identify recent clinical trials, reviews, editorials and meetings describing the biochemical and physiological role of individual micronutrients. No attempt was made to grade the evidence. The searches confirmed that the water-soluble vitamins (B group and C), together with the minerals, calcium, magnesium and zinc, are most relevant to cognitive performance. Clinical evidence revealed that marginal deficiencies of one or more of these micronutrients are not uncommon, even in the developed countries, and that such deficiencies may affect cognitive performance, especially in vulnerable groups such as the elderly and those individuals who are exposed to occupational pressures and a stressful lifestyle.