Classroom as Genome: Using the Tools of Genomics and Bioinformatics to Illuminate Classroom Observation Data.
Classroom observation protocols can provide an exceedingly rich form of data. However, this is a double-edged sword, as researchers often struggle to take full advantage of the detailed data outputs. In this essay, we introduce a new approach to the analysis of classroom observation data, termed "classroom as genome" (CAG). We illustrate how real-time classroom observation data and genomic data can be viewed as quite analogous, both conceptually and in terms of downstream analysis. We provide both abstract and concrete examples of how the tools of genomics and bioinformatics can be applied to classroom observation outputs. We also show how this philosophy of analysis allows for the layering of information from multiple observation protocols onto the same classroom data. The CAG approach enables biology education researchers to explore detailed patterns within observed classrooms in a highly scalable manner.


Phospholipase A2 enzymes, high-dose atorvastatin, and prediction of ischemic events after acute coronary syndromes.
Secretory phospholipase A2 (sPLA(2)) and lipoprotein-associated phospholipase A2 (Lp-PLA(2)) are enzyme biomarkers of increased cardiovascular risk and targets of emerging therapeutic agents. Their relationship to cardiovascular events in the setting of high-dose statin therapy compared with placebo in patients with acute coronary syndrome is not known. sPLA(2) and Lp-PLA(2) mass and activity were measured in 2587 patients in the Myocardial Ischemia Reduction With Acute Cholesterol Lowering (MIRACL) trial at baseline and after 16 weeks of treatment with atorvastatin 80 mg/d or placebo. Baseline levels of sPLA(2) and Lp-PLA(2) mass and activity were not associated with the primary efficacy measure of the trial of death, myocardial infarction, or unstable angina. However, in the overall cohort, baseline sPLA(2) mass predicted risk of death after multivariable adjustment (hazard ratio for 2-fold increase, 1.30; 95% confidence interval, 1.09-1.56; P=0.004). This association remained significant when examined separately in the placebo group but not in the atorvastatin group. Compared with placebo, atorvastatin reduced median sPLA(2) mass (-32.1% versus -23.1%), sPLA(2) activity (-29.5% versus -19.2%), Lp-PLA(2) mass (-35.8% versus -6.2%), and Lp-PLA(2) activity (-24.3% versus 5.4%; P<0.001 for all). Atorvastatin reduced the hazard of death associated with elevated sPLA(2) mass and activity by ≈50%. sPLA(2) mass independently predicts death during a 16-week period after acute coronary syndrome. High-dose atorvastatin significantly reduces sPLA(2) and Lp-PLA(2) mass and activity after acute coronary syndrome and mitigates the risk of death associated with sPLA(2) mass. Atorvastatin may exert antiinflammatory effects on phospholipases that contribute to its therapeutic benefit after acute coronary syndrome.


Medical students talking to patients.
Observations were made of 108 pre-clinical medical students interviewing patients carefully selected by general practitioner tutors, under strict supervision in an audio-visual studio. Videotape replay allowed immediate feed-back to the student, and this was supplemented by direct comment from the patient. The consultations were also used for small group teaching (by CCTV). Although most students viewed the exercise as threatening, they appeared to find it a helpful experience. The method uncovered students with major defects in communicating at the personal level. Detailed analyses of the content of the interviews are being carried out, and will be the subject of a separate communication.


Altered expression of ADAMs (A Disintegrin And Metalloproteinase) in fibrillating human atria.
ADAMs (A Disintegrin And Metalloproteinase) are ectoproteases that have recently been reported to be expressed in cardiac tissue. Although they are known to regulate cell-cell and cell-matrix interactions, their pathophysiological role in various cardiac diseases is unclear. The purpose of the present study was to determine whether structural remodeling of the atria during atrial fibrillation (AF) is associated with altered ADAM expression. Atrial tissue samples of 30 patients undergoing open-heart surgery were examined. Fifteen patients had persistent AF (> or =6 months), and the remaining 15 patients had no history of AF. ADAM9, ADAM10, and ADAM15 expression was analyzed quantitatively at the mRNA and protein levels. ADAM expression was localized by immunohistochemistry. ADAM expression was correlated with amounts of integrins beta1 and beta3. The amount of ADAM10 protein more than doubled during AF (82+/-15 versus 36+/-8 U; P<0.01). Amounts of ADAM15 protein (102+/-12 versus 40+/-6 U; P<0.01) and mRNA (24.0+/-5.6 versus 10.5+/-2.5 U; P<0.05) increased significantly during AF compared with sinus rhythm. ADAM9 protein was not detected in any sample. ADAM/integrin ratios showed an increase of 4- to 6-fold (P<0.05) in patients with AF who had significantly dilated atria (4.94+/-0.6 versus 4.3+/-0.7 cm; P<0.05). ADAM/integrin ratios correlated with atrial diameter. AF is associated with an increase in the expression of ADAM10 and ADAM15. Enhanced ADAM-dependent disintegrin and metalloproteinase activity may be a molecular mechanism that contributes to the dilation of fibrillating human atria.


Rainbow smelt: the unusual case of cryoprotection by sustained glycerol production in an aquatic animal.
Rainbow smelt flourish at -1.8 °C, the freezing point of sea water. An antifreeze protein contributes to freeze point depression but, more importantly, cryoprotection is due to an elevation in osmotic pressure, by the accumulation of glycerol. The lower the water temperature, the higher the plasma glycerol with levels recorded as high as 400 mmol l(-1). Glycerol freely diffuses out in direct relation to the glycerol concentration and fish may lose as much as 15% of their glycerol reserve per day. Glycerol levels decrease from a maximum in February/March while water temperature is still sub-zero. The decrease in glycerol may respond to a photoperiod signal as opposed to initiation which is triggered by low temperature. The initial increase in glycerol level is supported by liver glycogen but high sustained glycerol level is dependent upon dietary carbohydrate and protein. The metabolic pathways leading to glycerol involve flux from glycogen/glucose to the level of dihydroxyacetone phosphate (DHAP) via the initial part of glycolysis and from amino acids via a truncated gluconeogenesis again to the level of DHAP. DHAP in turn is converted to glycerol 3-phosphate (G3P) and then directly to glycerol. The key to directing DHAP to G3P is a highly active glycerol 3-P dehydrogenase. G3P is converted directly to glycerol via G3P phosphatase, the rate-limiting step in the process. The transition to glycerol production is associated with increased activities of enzymes at key loci in the top part of glycogenolysis/glycolysis. Curtailment of the final section of glycolysis may reside at the level of pyruvate oxidation with an inactivation of pyruvate dehydrogenase (PDH) driven by increased levels of PDH kinase. Enzymes associated with amino acid trafficking are elevated as is the pivotal enzyme phosphoenolpyruvate carboxykinase.


Identification of subgroups in the Danish population for targeted human papillomavirus vaccination efforts.
In the Danish childhood vaccination program, the human papillomavirus (HPV) vaccination coverage is lower than for other vaccines. To tailor a targeted HPV vaccination effort, we aimed to identify girls in Denmark with lower first dose HPV vaccination coverage than girls in general. A population-based retrospective cohort study was performed of girls born in 2001-2004, residing in Denmark in September 2019 (N = 128,351). Data from the Danish Vaccination Register was linked to sociodemographic data from the Danish Civil Registration System and Statistics Denmark. Cox's proportional hazard regression models were used to compare vaccination uptake rates between subgroups of girls. HPV vaccination coverage at 14 years of age varied widely by municipality (53.4-80.6%). Girls living with neither of their parents had a lower chance of being vaccinated compared to girls living with both their parents (HR 0.43; 95% CI 0.41-0.46), likewise for girls attending special need education compared with girls attending public schools (HR 0.50; 95% CI 0.42-0.59). The vaccination uptake among immigrants was lower compared to Danish-born girls (HR 0.51; 95% CI 0.49-0.54), especially among immigrant girls whose parents had not passed any Danish exams. Finally, girls who were DTaP-IPV revaccinated had a 50% greater chance of being HPV vaccinated compared to girls who were not (HR 1.61; 95% CI 1.58-1.64). To increase the HPV vaccination uptake, we recommend vaccination efforts targeting girls living without any of their parents, girls attending special need education, immigrants, and girls who are not DTaP-IPV revaccinated. When targeting immigrants, the effort should focus on disseminating sufficient and understandable information about the Danish childhood vaccination program to the parents.


An emergency department radiology audit procedure.
An audit was done to ascertain the concordance of radiographic interpretation by the emergency physician and the radiologist. In 8,021 radiographic procedures during the two-month study, concordance of interpretation was found in 97.6%. The primary area of interpretation difficulty was chest radiographs. Specifically, there was limited agreement (10 of 45 films, 22%) regarding the reading of inflammatory processes. Despite this, 40 of the 45 patients (89%) whose chest radiographs were reviewed received proper therapy. The findings in five radiographs were deemed to be significant enough to possibly lead to changes in management.


Volume conductor models in surface electromyography: applications to signal interpretation and algorithm test.
Simulation of surface electromyogram (EMG) has provided support for the development and testing of algorithms (e.g., for the extraction of amplitude or spectral properties of the EMG or for the estimation of the muscle fiber conduction velocity) and for the interpretation of experimental results. This is the second part of a pair of papers: the first explains the methods for the development of structure based models of surface EMG; this one provides a summary of interesting applications for the interpretation of signals and testing of algorithms. Simulations from some models already introduced in the literature are compared. Moreover, estimation of indexes from an innovative layered volume conductor including a subcutaneous tissue with variable thickness in space is studied.


Gene amplification and splice variants of 25-hydroxyvitamin D3 1,alpha-hydroxylase (CYP27B1) in glioblastoma multiforme--a possible role in tumor progression?
We report a gene amplification of 25-hydroxyvitamin D3 1,alpha-hydroxylase and the appearance of various splice variants in glioblastoma multiforme. Some protein products of the widespread expressed variants may exert a biological function. The analysis of amplification and overexpression of alternative transcripts will provide new insights in the regulation of the 25-hydroxyvitamin D3 1,alpha-hydroxylase expression and may have implications for prognosis in glioblastoma.


13C discrimination during CO2 assimilation by the terrestrial biosphere.
Estimates of the extent of the discrimination against13CO2 during photosynthesis (ΔA) on a global basis were made using gridded data sets of temperature, precipitation, elevation, humidity and vegetation type. Stomatal responses to leaf-to-air vapour mole fraction difference (D, leaf-to-air vapour pressure difference divided by atmospheric pressure) were first determined by a literature review and by assuming that stomatal behaviour results in the optimisation of plant water use in relation to carbon gain. Using monthly time steps, modelled stomatal responses toD were used to calculate the ratio of stomatal cavity to ambient CO2 mole fractions and then, in association with leaf internal conductances, to calculate ΔA. Weighted according to gross primary productivity (GPP, annual net CO2 asimilation per unit ground area), estimated ΔA for C3 biomes ranged from 12.9‰ for xerophytic woods and shrub to 19.6‰ for cool/cold deciduous forest, with an average value from C3 plants of 17.8‰. This is slightly less than the commonly used values of 18-20‰. For C4 plants the average modelled discrimination was 3.6‰, again slightly less than would be calculated from C4 plant dry matter carbon isotopic composition (yielding around 5‰). From our model we estimate that, on a global basis, 21% of GPP is by C4 plants and for the terrestrial biosphere as a whole we calculate an average isotope discrimination during photosynthesis of 14.8‰. There are large variations in ΔA across the globe, the largest of which are associated with the precence or absence of C4 plants. Due to longitudinal variations in ΔA, there are problems in using latitudinally averaged terrestrial carbon isotope discriminations to calculate the ratio of net oceanic to net terrestrial carbon fluxes.


Adjunctive analgesic therapy in veterinary medicine.
Adjunctive analgesic therapies are interventions for pain that involve agents or techniques other than the traditional analgesics (opioids, nonsteroidal anti-inflammatory drugs, and local anesthetics). Adjunctive therapies may be pharmacologic or nonpharmacologic in nature. The focus of this article is on pharmacologic interventions with potential utility as adjunctive analgesics in veterinary medicine. Pharmacology of selected agents, including medetomidine, ketamine, amantadine, gabapentin, systemic lidocaine, and pamidronate, is discussed in addition to evidence for their safety and efficacy and guidelines for their use in veterinary patients.


Optimization of psychopharmacotherapy for schizophrenia in a male, locked, non-acute unit serving for persistently ill patients over one year.
We describe real-world psychopharmacological treatment in a Japanese, male, closed psychiatric unit where clozapie was still unavailable. Fifty-five persistently-ill patients with schizophrenia (ICD-10), mean ± S.D. age: 57.5 ± 13.0 y.o., duration of illness and admissions: 30.9 ± 15.2 years and 20.7 ± 14.5 years, respectively) treated longitudinally were evaluated. The rule was to treat with a simplest possible psychotropic regimen without polypharmacy. Compared to the baseline, the number and dose of antipsychotics were reduced from 1.9 to 1.1 and 1012 mg/day to 607 mg/day, respectively. The number of total psychotropics was minimized from 4.7 to 2.1, with a simplified once or twice daily dosing. Overall, the CGI-Severity and FACT-Sz (global functioning) improved slightly from 5.8 to 5.5 and 28.7 to 32.6, respectively. Of note, no patients got worse in comparison with the baseline clinical presentation. Forty-four patients were successfully treated with a single antipsychotic; only seven needed two antipsychotics simultaneously while 36 had been treated with antipsychotic polypharmacy at baseline. Benzodiazepines (mostly lorazepam) and antiparkinsonian drugs were prescribed in 28 and only two, respectively. Nineteen needed adjunctive valproate (average blood levels: 99.3 ± 21.8 μg/mL) and nine used lithium (0.61 ± 0.26 mEq/L). Optimization of psychopharmacotherapy is still possible for difficult-to-treat patients and, while augmentation of an antipsychotic with mood stabilizers is frequently needed, antipsychotic polypharmacy should be exceptional.


Hidradenitis suppurativa (acne inversa): management of a recalcitrant disease.
Hidradenitis suppurativa is a chronic relapsing disorder of follicular occlusion that is often recalcitrant to therapy. Topical and systemic antibiotics, hormonal therapies, oral retinoids, immunosuppressant agents, and surgical treatment are some of the therapeutic alternatives used for this often recalcitrant and frequently troublesome disorder. This article reviews the pathophysiology of hidradenitis suppurativa, an evidence-based analysis of standard treatments, and recent advances in the therapy of this disorder.


Electrochemical sandwich-type thrombin aptasensor based on dual signal amplification strategy of silver nanowires and hollow Au-CeO2.
Signal amplification is crucial in electrochemical biosensor to obtain low detection limits. In this work, a highly sensitive sandwich-type thrombin aptasensor is constructed, based on dual signal amplification of uniform silver nanowires (AgNWs) and hollow Au-CeO2 nanocomposites. AgNWs are decorated on the ITO surface to immobilize amino functionalized thrombin capture apemeter 1 (Apt1). And Au nanoparticles (AuNPs) grown directly on the surface of hollow CeO2 microstructure are used to immobilize sulfydryl functionalized thrombin reporter apemeter 2 (Apt2). Thus, sandwich-type apatasensor has been successfully designed, due to the specific recognition between thrombin and the two kinds of aptamers. One of the signal amplifications is from the good conductivity of uniform AgNWs. Moreover, uniform AgNWs together with hollow Au-CeO2 exhibit the good catalytic performance for the reduction of H2O2, further resulting in significant electrochemical signal amplification. Because the electrochemical signal amplification is closely related to the thrombin concentration, differential pulse voltammetry is used to specifically detect thrombin. Under the optimized conditions, the proposed method has a good linear response ranged from 0.5 pM to 30 nM with a low detection limit of 0.25 pM (S/N = 3) for thrombin. The proposed thrombin aptasensor displays good selectivity, reproducibility and stability, providing a good platform for the ultrasensitive detection of thrombin.


Peptidomic analysis reveals proteolytic activity of kefir microorganisms on bovine milk proteins.
The microorganisms that make up kefir grains are well known for lactose fermentation, but the extent to which they hydrolyze and consume milk proteins remains poorly understood. Peptidomics technologies were used to examine the proteolytic activity of kefir grains on bovine milk proteins. Gel electrophoresis revealed substantial digestion of milk proteins by kefir grains, with mass spectrometric analysis showing the release of 609 protein fragments and alteration of the abundance of >1500 peptides that derived from 27 milk proteins. Kefir contained 25 peptides identified from the literature as having biological activity, including those with antihypertensive, antimicrobial, immunomodulatory, opioid and anti-oxidative functions. 16S rRNA and shotgun metagenomic sequencing identified the principle taxa in the culture as Lactobacillus species. The model kefir sample contained thousands of protein fragments released in part by kefir microorganisms and in part by native milk proteases.


A2A and A2B adenosine receptors: The extracellular loop 2 determines high (A2A) or low affinity (A2B) for adenosine.
A2A and A2B adenosine receptors (ARs) are closely related G protein-coupled receptor subtypes, which represent important (potential) drug targets. Despite their almost identical binding sites for adenosine, A2AARs are activated by low (nanomolar) adenosine concentrations, while A2BARs require micromolar concentrations. In the present study, we exchanged the extracellular loop 2 (ECL2) of the human A2AAR for that of the A2BAR. The resulting chimeric A2A(ECL2-A2B)AR was investigated in radioligand binding and cAMP accumulation assays in comparison to the wildtype A2AAR. While the ribose-modified adenosine analog N-ethylcarboxamidoadenosine (NECA) and its 2-substituted derivative CGS-21680 did not exhibit significant changes, adenosine showed dramatically reduced potency and affinity for the A2A(ECL2-A2B)AR mutant displaying similarly low potency as for the wt A2BAR. Supervised molecular dynamics simulation studies predicted a meta-binding site with high affinity for adenosine, but not for NECA, which may contribute to the observed effects.


Advances in microRNA experimental approaches to study physiological regulation of gene products implicated in CNS disorders.
The central nervous system (CNS) is a remarkably complex organ system, requiring an equally complex network of molecular pathways controlling the multitude of diverse, cellular activities. Gene expression is a critical node at which regulatory control of molecular networks is implemented. As such, elucidating the various mechanisms employed in the physiological regulation of gene expression in the CNS is important both for establishing a reference for comparison to the diseased state and for expanding the set of validated drug targets available for disease intervention. MicroRNAs (miRNAs) are an abundant class of small RNA that mediates potent inhibitory effects on global gene expression. Recent advances have been made in methods employed to study the contribution of these miRNAs to gene expression. Here we review these latest advances and present a methodological workflow from the perspective of an investigator studying the physiological regulation of a gene of interest. We discuss methods for identifying putative miRNA target sites in a transcript of interest, strategies for validating predicted target sites, assays for detecting miRNA expression, and approaches for disrupting endogenous miRNA function. We consider both advantages and limitations, highlighting certain caveats that inform the suitability of a given method for a specific application. Through careful implementation of the appropriate methodologies discussed herein, we are optimistic that important discoveries related to miRNA participation in CNS physiology and dysfunction are on the horizon.


The contribution of corporate initiatives to global renewable electricity deployment.
Climate change is gaining importance on the agenda of senior decision makers in the private sector. Hence, corporate renewable electricity (RE) procurement may become more relevant to the energy transition. RE100 is the largest corporate initiative to foster RE procurement with 315 corporate members as of 2021. Yet, the contribution of such initiatives to the energy transition remains unclear, because public reporting is aggregated on the global level. Here, we develop an approach to map the electricity procured by RE100 companies to jurisdictions worldwide, which allows estimating whether and where RE100 can have a transformative impact. We find that these companies source electricity in 129 jurisdictions, accounting for <1% of total electricity generation (RE and non-RE), thus dampening the hopes about the impact of RE100 on the global energy transition. RE100 companies procure 1.4% of available RE, exceeding 20% in nine jurisdictions. To increase its impact, RE100 should focus on interim targets and expansion. By 2030, stringent and frequent interim targets could lead to a cumulated additional 361 TWh of RE procured by RE100 companies, and a realistic membership expansion could lead to procurement of 7.7% of globally available RE by RE100 companies.


Evaluation of highly polar ionic liquid gas chromatographic column for the determination of the fatty acids in milk fat.
The SLB-IL111, a new ionic liquid capillary column for gas chromatography available from Supelco Inc., was recently shown to provide enhanced separation of unsaturated geometric and positional isomers of fatty acid (FAs) when it was compared to cyanopropylsiloxane (CPS) columns currently recommended for the analysis of fatty acid methyl esters (FAMEs). A 200 m SLB-IL111 capillary column, operated under a combined temperature and eluent flow gradient, was successfully used to resolve most of the FAs contained in milk fat in a single 80 min chromatographic separation. The selected chromatographic conditions provided a balanced, simultaneous separation of short-chain (from 4:0), long-chain polyunsaturated fatty acids (PUFAs), and most of the unsaturated FA positional/geometric isomers contained in milk fat. Among the monounsaturated fatty acids (MUFAs), these conditions separated t11-18:1 and t10-18:1 FAs, the two most abundant trans fatty acids (t-FA) contained in most dairy products. These t-FAs reportedly have different biological activities. The conjugated linoleic acid (CLA) isomers commonly found in dairy products were separated from each other, including t7,c9-18:2 from c9,t11-18:2, which eliminated the need for their complementary silver ion HPLC analysis. The application of the SLB-IL111 column provided a complementary elution profile of FAMEs to those obtained by CPS columns, allowing for a more comprehensive FA analysis of total milk fat. The FAMEs were identified by the use of available reference materials, previously synthesized and characterized reference mixtures, and prior separations of the milk fat FAMEs by silver ion chromatography based on the number/geometry of double bonds.


Type of feeding does not influence thyroid hormone levels in premature infants.
Thirty premature neonates (mean gestational age 31.8 +/- 3.90 weeks; mean birth weight 1569 +/- 236 g) were divided into three groups receiving breast milk, Robolact and Pre-Aptamil formula feeding. The infants with and without neonatal idiopathic respiratory distress syndrome (IRDS) were separately evaluated. No effect of the type of feeding and of a previously experienced IRDS on the blood T3-, T4-, and THS-levels could be demonstrated at the age of 6 weeks.


Ezetimibe--a new approach in hypercholesterolemia management.
Ezetimibe is the first agent used in hypercholesterolemia treatment known to lower intestinal cholesterol uptake that is able to inhibit NPC1L1 transport proteins in the brush boarder of enterocytes and macrophages. Furthermore, it demonstrates anti-inflammatory and immunomodulatory properties and influences the expression of certain antigens. The drug is rapidly absorbed from the gastrointestinal tract and is then glucuronidated to form the active metabolite. It also undergoes extensive enterohepatic circulation. Various genetic polymorphisms seem to influence the pharmacokinetics of ezetimibe with different effects. The drug also presents a complex impact on cytochrome P450 enzymes, as it is a metabolism-dependent inhibitor of CYP3A4. Ezetimibe does not demonstrate any clinically significant interactions with statins, fibrates, mipomersen sodium, levothyroxine or lopinavir. However, its effect in conjunction with cyclosporine is not neutral. The use of this cholesterol absorption inhibitor has been shown to be safe and effective among patients after cardiac, renal and liver transplants, as well as in HIV patients.


Primary amyloidosis of urinary bladder and ureter: cause of mural calcification.
Primary amyloidosis is seen in the absence of predisposing disease. It may affect most organs in the body, including the collecting system of the urinary tract. Submucosal calcifications have been described in the renal pelvis and may be pathognomonic for primary amyloidosis of the renal pelvis. We have been able to demonstrate similar submucosal calcifications in the ureter and bladder. The differential diagnosis of submucosal calcification of the distal ureter and bladder must include bilharziasis and tuberculosis. With the recognition of submucosal calcifications, diagnosis of the underlying disease may be readily made. An illustrative case is presented.


Effect of E23G/K, F36V, N37T, E119D, and E199G polymerase acidic protein substitutions on the replication and baloxavir susceptibility of influenza B viruses.
Baloxavir marboxil (baloxavir) is a highly effective, single-dose influenza therapeutic. Identification of molecular markers in the target polymerase acidic (PA) protein that can diminish baloxavir efficacy is an ongoing goal of the scientific community. In this study, we generated recombinant Victoria-lineage and Yamagata-lineage influenza B viruses (IBVs) containing 6 substitutions (E23G/K, F36V, N37T, E119D, and E199G) spanning the endonuclease domain of the PA. Although 5 of these PA substitutions negatively impacted in vitro polymerase activity and replication kinetics, particularly in the Victoria-lineage IBV background, viruses with E119D exhibited activity levels comparable to those of wild-type viruses. Moreover, only E119D moderately decreased the susceptibility of IBVs to baloxavir (resulting in ∼2.0-fold to 2.6-fold elevated EC50s); viruses with the other substitutions exhibited normal drug inhibition. These results show that E119D may reduce the baloxavir susceptibility of IBVs without compromising their replicative fitness. Overall, this study expands the molecular landscape of PA substitutions potentially affecting baloxavir efficacy against IBV.


Depression and cardiovascular disease.
Approximately one out of every five patients with cardiovascular disease (CVD) suffers from major depressive disorder (MDD). Both MDD and depressive symptoms are risk factors for CVD incidence, severity and outcomes. Great progress has been made in understanding potential mediators between MDD and CVD, particularly focusing on health behaviors. Investigators have also made considerable strides in the diagnosis and treatment of depression among patients with CVD. At the same time, many research questions remain. In what settings is depression screening most effective for patients with CVD? What is the optimal screening frequency? Which therapies are safe and effective? How can we better integrate the care of mental health conditions with that of CVD? How do we motivate depressed patients to change health behaviors? What technological tools can we use to improve care for depression? Gaining a more thorough understanding of the links between MDD and heart disease, and how best to diagnose and treat depression among these patients, has the potential to substantially reduce morbidity and mortality from CVD.


The capnogram as a guide to the efficacy of cardiac massage.
Recordings from three patients in cardiac arrest are shown to illustrate the importance of monitoring pulmonary perfusion by means of the capnogram as a continuous guide to the cardiac output achieved by cardiac massage and resuscitation. Such monitoring allows the performance to be adjusted to obtain the maximal result.


Metastatic brain tumor following negative second-look operation for ovarian carcinoma.
Three patients with stage III ovarian cancer had brain metastases following aggressive chemotherapy with Platinol and Adriamycin, and negative second-look surgery. The possibility of brain metastasis following complete control of intraperitoneal ovarian disease should be considered in patients with neurological symptoms.


Toll-like receptor 4 activation on human amniotic epithelial cells is a risk factor for pregnancy loss.
Maternal-fetal tolerance plays a fundamental role in the maintenance of pregnancy. However, this immunological tolerance can be influenced by intrauterine infections. Human amniotic epithelial cells (hAECs) have immunomodulatory effects and respond to invading pathogens through expressing various toll-like receptors (TLRs). We hypothesize that bacteria or bacterial products affect the immunosuppressive effects of hAECs through TLR stimulation. Here, we investigated how a successful pregnancy can be threatened by TLR4 activation on hAECs on lipopolysaccharide (LPS) engagement. hAECs were isolated from the amniotic membrane received from six healthy pregnant women. The immunophenotyping of hAECs was studied by flow cytometry. The isolated hAECs (4 × 105 cells/ml) were cultured in 24-well plates in the presence or absence of LPS (5 μg/ml). After 24, 48, and 72 h of incubation, the culture supernatants of hAECs were collected, and the levels of interleukin-5 (IL-5), IL-6, IL-1β, tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta 1 (TGF-β1), and prostaglandin E2 (PGE2) were measured by enzyme-linked immunosorbent assay. TLR4 activation showed a stimulatory effect on TGF-β1 production of hAECs (P < 0.001-0.05). PGE2 production of LPS-stimulated hAECs was significantly increased (P < 0.01-0.05). Moreover, TLR4 could induce TNF-α and IL-1β production of hAECs (P < 0.0001-0.01), while this effect was not observed on IL-6 production of hAECs. The IL-5 was produced at a very low level in two culture supernatants of hAECs, in which its production was independent of LPS effect. TLR4 activation by bacterial components on hAECs may be a potential risk factor for pregnancy complications.


The prevalence of binge eating disorder and borderline personality symptomatology among gastric surgery patients.
In this study, we examined the prevalence of binge eating disorder (BED) and borderline personality disorder (BPD) in a sample of 121 candidates seeking surgery for obesity. In this predominantly female sample (85.9%), according to the Questionnaire on Eating and Weight Patterns-Revised (QEWP-R), the prevalence of BED was 6.5%. As for the prevalence of BPD, 14.0% exceeded the clinical cut-off score on the Self-Harm Inventory (SHI), 14.0% exceeded the clinical cut-off score on the borderline personality scale of the Personality Diagnostic Questionnaire-4 (PDQ-4), and 7.4% exceeded the clinical cut-off score on the McLean Screening Inventory for Borderline Personality Disorder (MSI-BPD). Overall, 24.8% of the sample exceeded the clinical cut-off on at least one measure of BPD whereas only 3.3% exceeded the clinical cut-off on all three measures. In addition, there was a significant inverse relationship between the discrepancy between highest and lowest adult body mass index, and scores on the PDQ-4 and the MSI-BPD. The authors discuss the implications of these findings.


Core-shell Fe3O4-Au magnetic nanoparticles based nonenzymatic ultrasensitive electrochemiluminescence immunosensor using quantum dots functionalized graphene sheet as labels.
In this paper, a novel, low-cost electrochemiluminescence (ECL) immunosensor using core-shell Fe3O4-Au magnetic nanoparticles (AuMNPs) as the carriers of the primary antibody of carbohydrate antigen 125 (CA125) was designed. Graphene sheet (GS) with property of good conductivity and large surface area was a captivating candidate to amplify ECL signal. We successively synthesized functionalized GS by loading large amounts of quantum dots (QDs) onto the poly (diallyldimethyl-ammonium chloride) (PDDA) coated graphene sheet (P-GS@QDs) via self-assembly electrostatic reactions, which were used to label secondary antibodies. The ECL immunosensors coupled with a microfluidic strategy exhibited a wide detection range (0.005-50 U mL(-1)) and a low detection limit (1.2 mU mL(-1)) with the help of an external magnetic field to gather immunosensors. The method was evaluated with clinical serum sample, receiving good correlation with results from commercially available analytical procedure.


Fabrication of homogenous multilayered W films by multi-step sputtering deposition: a novel grain boundary enrichment strategy.
Using a multi-step deposition approach, we develop a strategy of homogeneous multilayered (HM) structure to enrich the grain boundary (GB) of sputtered W films. In comparison with the single-layered film, the HM W film is easily controllable for the film GB density. When decreasing the film modulation period t m from 160 nm to 7 nm, the GB density gradually increased from 0.065 nm(-1) to 0.275 nm(-1) without changing the phase structure of the films. Accordingly, the film's electrical resistivity and mechanical hardness, which are related to the GBs, changed from 40.1 μΩ · cm to 75.3 μΩ · cm and from 12.1 GPa to 16.2 GPa, respectively. Detailed analysis showed that the formation of an HM structure is related to the temperature evolution of the film growing surface during the multi-step sputtering process. This study could provide a general engineering approach to enrich film interfaces and allows for the development of thin films with novel microstructures.


Platelet extracellular vesicles in COVID-19: Potential markers and makers.
Platelets and platelet extracellular vesicles (pEV) are at the crossroads of coagulation and immunity. Extracellular vesicles are messengers that not only transmit signals between cells, but also provide information about the status of their cell of origin. Thus, pEVs have potential as both biomarkers of platelet activation and contributors to pathology. Coronavirus Disease-19 (COVID-19), caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a complex disease affecting multiple organs and is characterized by a high degree of inflammation and risk of thrombosis in some patients. In this review, we introduce pEVs as valuable biomarkers in disease with a special focus on their potential as predictors of and contributors to COVID-19.


Drug distribution in dog brain studied by positron emission tomography.
We used positron emission tomography to monitor the distribution of radioactivity in dog brain and muscle following i.v. administration of 11C-labelled antipyrine, imipramine, and quinidine. Twenty-five sequential scans of a transaxial slice of the head were performed within 90 min; radioactivity in plasma was measured in a gamma-counter. Following i.v. injection of [11C]antipyrine (50 mg kg-1; 9-68 mCi; n = 10), the decay of plasma activity was accompanied by rapid uptake in brain and variable uptake in muscle, immediately followed by a redistribution leading to equalization of the radioactivity in the tissues. Administration of [11C]imipramine (4 mg kg-1; 30-110 mCi; n = 8) was followed by a rapid build-up of a sustained gradient between high brain, and low plasma and muscle radioactivity. After i.v. injection of [11C]quinidine (1 mg kg-1; 11-87 mCi; n = 10), radioactivity in brain was low, with higher activity in plasma and muscle throughout the experiment. Positron emission tomography thus revealed for each drug a distinct pattern of distribution consistent with established properties of the compounds. This technique seems promising for the study of early drug distribution, notwithstanding certain limitations.


Inactivation of the Wip1 phosphatase inhibits mammary tumorigenesis through p38 MAPK-mediated activation of the p16(Ink4a)-p19(Arf) pathway.
Modulation of tumor suppressor activities may provide new opportunities for cancer therapy. Here we show that disruption of the gene Ppm1d encoding Wip1 phosphatase activated the p53 and p16 (also called Ink4a)-p19 (also called ARF) pathways through p38 MAPK signaling and suppressed in vitro transformation of mouse embryo fibroblasts (MEFs) by oncogenes. Disruption of the gene Cdkn2a (encoding p16 and p19), but not of Trp53 (encoding p53), reconstituted cell transformation in Ppm1d-null MEFs. In vivo, deletion of Ppm1d in mice bearing mouse mammary tumor virus (MMTV) promoter-driven oncogenes Erbb2 (also called c-neu) or Hras1 impaired mammary carcinogenesis, whereas reduced expression of p16 and p19 by methylation-induced silencing or inactivation of p38 MAPK correlated with tumor appearance. We conclude that inactivation or depletion of the Wip1 phosphatase with resultant p38 MAPK activation suppresses tumor appearance by modulating the Cdkn2a tumor-suppressor locus.


Met(104) is the CO-replaceable ligand at Fe(II) heme in the CO-sensing transcription factor BxRcoM-1.
Both Met(104) and Met(105) are involved, either directly or indirectly, in the redox mediated ligand switch of the heme-dependent transcription factor, RcoM-1. Recent studies of Burkholderia xenovorans RcoM identified Cys(94) as the thiolate ligand in the Fe(III) state of the heme cofactor. Upon reduction, a neutral donor replaces Cys(94) trans to His(74). Homology modelling implicated either Met(104) or Met(105) as the possible ligand in the Fe(II) state. We spectroscopically compared wild type (WT) RcoM-1 to three Met-to-Leu variants (M104L, M105L, and M104L/M105L) to identify which Met residue acts as the ligand. All proteins were isolated as admixtures of Fe(III) and Fe(II)-CO heme; oxidation by ferricyanide enables study of homogeneous oxidation and coordination states. Met(104) is the CO-replaceable Fe(II) heme ligand. The magnetic circular dichroism (MCD) spectrum of Fe(II) M105L resembled WT. M104L and M104L/M105L, however, showed spectra arising from the formation of a high-spin, five-coordinate species indicating the loss of the ligand. The electron paramagnetic resonance (EPR) spectra of WT Fe(III) RcoM-1, oxidized Fe(III) M104L, and as-isolated M105L exhibited narrow, rhombic low-spin signals typical of thiolate-bound hemes. In contrast, oxidized Fe(III) M105L and oxidized Fe(III) M104L/M105L revealed a broad, rhombic low-spin, six-coordinate signal indicative of replacement of the thiolate by a neutral ligand. Thus, we conclude that Met(105) is important to the stability of the Fe(III) heme pocket during oxidation.


Flecainide-induced pneumonitis: a case report.
We report a case of acute respiratory distress associated with a histological pattern of acute fibrinous and organizing pneumonia, and discuss the possible responsibility of flecainide therapy. A 61-year-old African woman developed a rapidly progressive dyspnea and required admission in the intensive care unit for orotracheal intubation and mechanical ventilation. Chest X-ray examination revealed bilateral infiltrates predominating in the basal part of both lungs. Lung computed tomography disclosed bilateral ground-glass opacities and septal thickening. After exclusion of the most common causes of infectious or immune pneumonia, a toxic origin was investigated and flecainide toxicity was considered. Lung biopsy was consistent with the unusual pattern of acute fibrinous and organizing pneumonia. Clinical and radiological improvement was noted after corticosteroid therapy, but the patient died from septic complications. Flecainide-induced lung injury has rarely been reported in the literature and remains a diagnosis of exclusion. The histological pattern of acute fibrinous and organizing pneumonia has been previously observed with amiodarone. There are no firm guidelines for the treatment of acute fibrinous and organizing pneumonia, but some patients may positively respond to corticosteroids.


Expression of infectious pancreatic necrosis virus (IPNV) VP2-VP3 fusion protein in Lactobacillus casei and immunogenicity in rainbow trouts.
Infectious pancreatic necrosis virus (IPNV) infects wild and cultured salmonids, causing high mortality in juvenile trouts and salmons. IPNV VP2-VP3 fusion gene was constructed by splicing overlap extension (SOE) PCR and inserted into Lactobacillus/Escherichia coli shuttle vectors (pPG1and pPG2) followed by transformation of Lactobacillus casei competent cell to yield two recombinant strains: Lc:PG1-VP2-VP3 (surface-displayed) and Lc:PG2-VP2-VP3 (secretory). Subsequently, juvenile rainbow trouts were inoculated with the recombinant strains via orogastric route. Our results demonstrated that Lactobacillus-derived VP2-VP3 fusion protein could induce production of serum IgM specific for IPNV with neutralizing activity in rainbow trouts. Statistical analyses of IgM levels showed that immunogenicity of Lc:PG1-VP2-VP3 was more powerful than that of Lc:PG2-VP2-VP3 (P<0.001) in rainbow trouts. This result has been confirmed by viral loads reduction analyzed by real-time RT-PCR in orogastrically immunized rainbow trouts after virus challenging. Comparing to trouts received Lactobacillus (control), rainbow trouts orogastrically dosed with Lc:PG1-VP2-VP3 resulted in ∼10-fold reduction in viral loads on day 10 post-virus challenging, and ∼4-fold did by Lc:PG2-VP2-VP3. Taken together, Lc:PG1-VP2-VP3 functions as novel mucosal vaccine against IPNV infection in rainbow trouts, which most likely come true.


Outcomes of patients with pre-existing disability managed by mobile stroke units: A sub-analysis of the BEST-MSU study.
Few data exist on acute stroke treatment in patients with pre-existing disability (PD) since they are usually excluded from clinical trials. A recent trial of mobile stroke units (MSUs) demonstrated faster treatment and improved outcomes, and included PD patients. To determine outcomes with tissue plasminogen activator (tPA), and benefit of MSU versus management by emergency medical services (EMS), for PD patients. Primary outcomes were utility-weighted modified Rankin Scale (uw-mRS). Linear and logistic regression models compared outcomes in patients with versus without PD, and PD patients treated by MSU versus standard management by EMS. Time metrics, safety, quality of life, and health-care utilization were compared. Of the 1047 tPA-eligible ischemic stroke patients, 254 were with PD (baseline mRS 2-5) and 793 were without PD (baseline mRS 0-1). Although PD patients had worse 90-day uw-mRS, higher mortality, more health-care utilization, and worse quality of life than non-disabled patients, 53% returned to at least their baseline mRS, those treated faster had better outcome, and there was no increased bleeding risk. Comparing PD patients treated by MSU versus EMS, 90-day uw-mRS was 0.42 versus 0.36 (p = 0.07) and 57% versus 46% returned to at least their baseline mRS. There was no interaction between disability status and MSU versus EMS group assignment (p = 0.67) for 90-day uw-mRS. PD did not prevent the benefit of faster treatment with tPA in the BEST-MSU study. Our data support inclusion of PD patients in the MSU management paradigm.


Genetic polymorphisms of glutathione S-transferase T1 (GSTT1) and M1 (GSTM1) in selected populations of Afghanistan.
Genetic polymorphisms in genes encoding glutathione S-transferase T1 (GSTT1, a member of class theta) and M1 (GSTM1, a member of class mu) have been defined. Previous studies have revealed that there was significant difference between populations for allelic frequency of several members of GSTs. In order to find the prevalence of null genotypes of GSTM1 and GSTT1 in Afghanis populations the present study was carried out. The total study subjects consisted of 656 unrelated healthy Afghanis refugees living in Fars province (southern Iran). From these 257, 217, 120, and 62 individuals were Pashtuns, Tajiks, Hazaras, and Uzbeks, respectively. Genetic polymorphisms for GSTT1 and GSTM1 were detected by multiplex PCR. The prevalence of null genotype of GSTM1 in Pashtuns, Tajiks, Hazaras, and Uzbeks was 42.4, 48.4, 52.5, and 40.3 %, respectively. There was no significant difference between these populations for the genotypic distribution of the GSTM1 polymorphism (χ(2) = 4.67, df = 3, P = 0.197). The frequency of GSTT1 null genotype in Pashtuns, Tajiks, Hazaras, and Uzbeks was 7.4, 25.3, 25.0, and 29.0 %, respectively. The observed difference between populations for prevalence of GSTT1 null genotype was statistically significant (χ(2) = 35.54, df = 3, P < 0.001). In comparison with European and Asian populations, Afghanistan populations like Iranian populations showed intermediate frequency for GSTT1 and GSTM1 null genotypes.


Small-scale distributional dynamics of the yellow water-lily and its herbivore Galerucella nymphaeae (Coleoptera: Chrysomelidae).
I followed the within-generation distribution of a chewing specialist herbivore, the water-lily beetle, on individually marked leaves of the yellow water-lily. Yellow water-lilies produced new leaves steadily throughout the growth season. Average leaf longevity was 3-4 weeks, much shorter than the developmental time of the beetle. The average egg-load of leaves was about 120. Leaf longevity was independent of egg density, but migration rate of the first instar larvae was density-dependent. Beetles occurred in every leaf, but consumed only a fraction (17%) of the available leaf area. However, this caused the leaf to lose its floating ability, so even this low rate of consumption made the leaf unavailable to herbivores. The herbivore population had to redistribute itself throughout the summer, escaping from the drowning leaves to fresh ones. No beetle could survive from egg to adult on a single leaf. The small-scale redistribution of the herbivore strongly affected the damage experienced by the host plant. In general, ability to redistribute depends on the dispersal ability of the herbivore, and thus migration ability may strongly affect the plant-herbivore interaction.


Comparative evaluation of the treatment efficacy of suberoylanilide hydroxamic acid (SAHA) and paclitaxel in ovarian cancer cell lines and primary ovarian cancer cells from patients.
In most patients with ovarian cancer, diagnosis occurs after the tumour has disseminated beyond the ovaries. In these cases, post-surgical taxane/platinum combination chemotherapy is the "gold standard". However, most of the patients experience disease relapse and eventually die due to the emergence of chemotherapy resistance. Histone deacetylase inhibitors are novel anticancer agents that hold promise to improve patient outcome. We compared a prototypic histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA), and paclitaxel for their treatment efficacy in ovarian cancer cell lines and in primary patient-derived ovarian cancer cells. The primary cancer cells were isolated from malignant ascites collected from five patients with stage III ovarian carcinomas. Cytotoxic activities were evaluated by Alamar Blue assay and by caspase-3 activation. The ability of SAHA to kill drug-resistant 2780AD cells was also assessed. By employing the cell lines OVCAR-3, SK-OV-3, and A2780, we established SAHA at concentrations of 1 to 20 microM to be as efficient in inducing cell death as paclitaxel at concentrations of 3 to 300 nM. Consequently, we treated the patient-derived cancer cells with these doses of the drugs. All five isolates were sensitive to SAHA, with cell killing ranging from 21% to 63% after a 72-h exposure to 20 microM SAHA, while four of them were resistant to paclitaxel (i.e., <10% cell death at 300 nM paclitaxel for 72 hours). Likewise, treatment with SAHA led to an increase in caspase-3 activity in all five isolates, whereas treatment with paclitaxel had no effect on caspase-3 activity in three of them. 2780AD cells were responsive to SAHA but resistant to paclitaxel. These ex vivo findings raise the possibility that SAHA may prove effective in the treatment of paclitaxel-resistant ovarian cancer in vivo.


New perspectives on the biology of fragile X syndrome.
Fragile X syndrome (FXS) is a trinucleotide repeat disorder caused by a CGG repeat expansion in FMR1, and loss of its protein product FMRP. Recent studies have provided increased support for the role of FMRP in translational repression via ribosomal stalling and the microRNA pathway. In neurons, particular focus has been placed on identifying the signaling pathways such as PI3K and mTOR downstream of group 1 metabotropic glutamate receptors (mGluR1/5) that regulate FMRP. New evidence also suggests that loss of FMRP causes presynaptic dysfunction and abnormal adult neurogenesis. In addition, studies on FXS stem cells especially induced pluripotent stem (iPS) cells and new sequencing efforts hold out promise for deeper understanding of the silencing process and mutation spectrum of FMR1.


Design, synthesis, and biological evaluation of a novel series of peripheral-selective noradrenaline reuptake inhibitors-Part 2.
Peripherally selective inhibition of noradrenaline reuptake is a novel mechanism for the treatment of stress urinary incontinence to overcome adverse effects associated with central action. Herein, we describe our medicinal chemistry approach to discover peripheral-selective noradrenaline reuptake inhibitors to avert the risk of P-gp-mediated DDI at the blood-brain barrier. We observed that steric shielding of the hydrogen-bond acceptors and donors (HBA and HBD) of compound 1 reduced the multidrug resistance protein 1 (MDR1) efflux ratio; however, the resulting compound 6, a methoxyacetamide derivative, was mainly metabolized by CYP2D6 and CYP2C19 in the in vitro phenotyping study, implying the risk of PK variability based on the genetic polymorphism of the CYPs. Replacement of the hydrogen atom with a deuterium atom in a strategic, metabolically hot spot led to compound 13, which was mainly metabolized by CYP3A4. To our knowledge, this study represents the first report of the effect of deuterium replacement for a major metabolic enzyme. The compound 13, N-{[(6S,7R)-7-(4-chloro-3-fluorophenyl)-1,4-oxazepan-6-yl]methyl}-2-[(2H(3))methyloxy]acetamide hydrochloride, which exhibited peripheral NET selective inhibition at tested doses in rats, increased urethral resistance in a dose-dependent manner.


An exploratory study on the effectiveness of "Calmare therapy" in patients with cancer-related neuropathic pain: A pilot study.
Calmare therapy (CT) has been suggested as a novel treatment for managing chronic pain. Recently, it was reported to show a positive therapeutic outcome for managing neuropathic pain condition. We performed an exploratory prospective study on the effectiveness of CT in patients with various types of cancer-related neuropathic pain (CNP). We performed an open-labeled, single-arm, exploratory study on the effectiveness of CT in patients with various types of cancer-related neuropathic pain (CNP). The primary endpoint was a comparison of the 11-point Numerical Rating Scale (NRS) pain score at one month with the baseline score in each patient. Brief Pain Inventory (BPI) and consumption of opioid were also evaluated during follow-up period. CT significantly decreased NRS pain score at one month from baseline (p < 0.001) in 20 patients with chemotherapy-induced peripheral neuropathy (n = 6), metastatic bone pain (n = 7), and post-surgical neuropathic pain (n = 7). It also improved overall BPI scores, decreased consumption of rescue opioid (p = 0.050), and was found satisfactory by a half of patients (n = 10, 50.0%). Our preliminary results suggest that CT may be considered for cancer patients with various types of CNP. Large studies are necessary to confirm our findings and ascertain which additional CNP show positive response to CT.


Peripheral GLP-1 gastroprotection against ethanol: the role of exendin, NO, CGRP, prostaglandins and blood flow.
The aim of this study was to investigate the effects of peripherally injected glucagon like peptide-1 (GLP-1) on ethanol-induced gastric mucosal damage and the mechanisms included in the effect. Absolute ethanol was administered through an orogastric cannula right after the injection of GLP-1 (1, 10, 100, 1000 or 10,000 ng/kg; i.p.). The rats were decapitated an hour later, the stomachs removed and the gastric mucosal damage scored. 1000 ng GLP-1 inhibited gastric mucosal damage by 45% and 10,000 ng GLP-1 by 60%. The specific receptor antagonist exendin-(9-39) (2500 ng/kg; i.p.), calcitonin gene related peptide (CGRP) receptor antagonist CGRP-(8-37) (10 microg/kg; i.p.), nitric oxide (NO) synthase inhibitor l-NAME (30 mg/kg; s.c.) and cyclooxygenase inhibitor indomethacin (5 mg/kg; i.p.) inhibited the preventive effect of GLP-1 on ethanol-induced gastric mucosal damage. GLP-1 also prevented the decrease in gastric mucosal blood flow caused by ethanol when administered at gastroprotective doses (1000 and 10,000 ng/kg; i.p.). In conclusion, GLP-1 administered peripherally prevents the gastric mucosal damage caused by ethanol in rats. CGRP, NO, prostaglandin and gastric mucosal blood flow are thought to play a role in this effect, mediated through receptors specific to GLP-1.


Therapeutic hypothermia for neonatal hypoxic-ischaemic encephalopathy had favourable outcomes at a referral hospital in a middle-income country.
This South African study documented the survival and neurodevelopmental outcomes of infants with hypoxic-ischaemic encephalopathy (HIE) after introducing cooling to a neonatal intensive care unit and identified early markers for neurodevelopmental outcome. We retrospectively reviewed infants that received cooling according to the Total Body Hypothermia trial protocol from 2008 to 2011. Infants were screened with the Bayley Scales of Infant and Toddler Development, Third Edition, at one year of age and underwent neurological and hearing assessments. Data on 99 infants with HIE showed that 45% of cases were moderate, 23% severe and 32% mild. An abnormal amplitude integrated electro-encephalogram (aEEG) background was documented in 45 cases within 24 hours. Magnetic resonance imaging (MRI) scans were consistent with HIE in all but one case. We reviewed 50 traceable survivors at one year. Development was significantly impaired in nine and 41 were normal or mildly impaired. A severely abnormal aEEG background, severe HIE and an abnormal MRI were associated with death and severe impairment. A good suck, mild HIE, primiparity and normal MRI were associated with good outcomes. Most infants with HIE survived without major impairment. Previously described predictors of neurodevelopmental outcome were good surrogate markers in this population.


P21-activated kinases in human cancer.
A balance between proliferation, differentiation, migration and death of cells is critical for the normal development of an organism. Perturbations of this balance can contribute to cancer development. The p21-activated serine/threonine kinases (Paks) play an important role in a variety of cellular functions including cell morphogenesis, motility, survival, angiogenesis, and mitosis. Paks were initially identified as an effector molecules of RHO GTPases; however, recent evidence that they can be activated in both GTPase-dependent and -independent manners expands our understanding of their physiologic functions. Paks play an important role in growth factor signaling, leading to cytoskeletal reorganization that subsequently influences growth factor-mediated cell migration and metastasis functions. Recent findings that Paks play a role in mitosis, nuclear receptor-signaling and deregulation of Pak in cancer cells suggest that these kinases play an important role in both normal development and cancer progression. In this review, we summarize the results of recent advances into the role of Paks in tumorigenesis and metastasis.


European Respiratory Society International Congress 2021: highlights from best-abstract awardees.
This article provides an overview of some of the highlights of the @EuroRespSoc Congress 2021 from the perspective of the best-abstract awardees of the ERS Assemblies @EarlyCareerERS @OrphaLung https://bit.ly/3JCjHYS.


Overcoming cloning problems by staining agarose gels with crystal violet instead of ethidium bromide in lactate dehydrogenase gene from Plasmodium vivax and Plasmodium falciparum.
In this study, lactate dehydrogenase gene from Plasmodium vivax has been tried to subclone into an expression vector. Some of the Plasmodium falciparum lactate dehydrogenase mutant genes have also been tried to clone and subclone into a vector, but we failed to clone or subclone either of the genes. DNA visualisation in electrophoretic gels typically requires UV radiation and the fluorecent dye ethidium bromide. A crystal violet-stained gel was run instead of an ethidium bromide gel and so avoided the use of UV radiation. This enabled us to clone or subclone both Plasmodium vivax lactate dehydrogenase gene and Plasmodium falciparum lactate dehydrogenase mutant genes into any desired vector.


PEAR1 regulates expansion of activated fibroblasts and deposition of extracellular matrix in pulmonary fibrosis.
Pulmonary fibrosis is a chronic interstitial lung disease that causes irreversible and progressive lung scarring and respiratory failure. Activation of fibroblasts plays a central role in the progression of pulmonary fibrosis. Here we show that platelet endothelial aggregation receptor 1 (PEAR1) in fibroblasts may serve as a target for pulmonary fibrosis therapy. Pear1 deficiency in aged mice spontaneously causes alveolar collagens accumulation. Mesenchyme-specific Pear1 deficiency aggravates bleomycin-induced pulmonary fibrosis, confirming that PEAR1 potentially modulates pulmonary fibrosis progression via regulation of mesenchymal cell function. Moreover, single cell and bulk tissue RNA-seq analysis of pulmonary fibroblast reveals the expansion of Activated-fibroblast cluster and enrichment of marker genes in extracellular matrix development in Pear1-/- fibrotic lungs. We further show that PEAR1 associates with Protein Phosphatase 1 to suppress fibrotic factors-induced intracellular signalling and fibroblast activation. Intratracheal aerosolization of monoclonal antibodies activating PEAR1 greatly ameliorates pulmonary fibrosis in both WT and Pear1-humanized mice, significantly improving their survival rate.


An algorithm to detect Chlamydia trachomatis by polymerase chain reaction on specimens extracted for enzyme immunoassay.
Amplification assays for detecting Chlamydia trachomatis have been shown to be more sensitive than enzyme immunoassay (EIA) by many investigators. In this study, we have developed an algorithm for performing PCR (COBAS AMPLICOR) on selected specimens extracted for EIA (ACCESS) with sample-to-cutoff (s/co) values between 0.25 and 0.99. Furthermore, we have shown that these specimens can be utilized for PCR without encountering any inhibition problems. In our investigation, 230 out of 6,558 urethral and cervical swabs submitted for C. trachomatis screening by EIA over a period of 9 months, had s/co values ranging between of 0.25 and 0.99. Ninety (39.1%) of these specimens tested positive by PCR. These specimens were stable and gave reproducible PCR results before and after storage for a period of 9 months. This testing algorithm offers an effective way of detecting C. trachomatis with selective use of PCR while increasing the sensitivity of the EIA screening system.


Interleukin-33 Exacerbates IgA Glomerulonephritis in Transgenic Mice Overexpressing B Cell Activating Factor.
The cytokine IL-33 is an activator of innate lymphoid cells 2 (ILC2s) in innate immunity and allergic inflammation. B cell activating factor (BAFF) plays a central role in B cell proliferation and differentiation, and high levels of this protein cause excess antibody production, including IgA. BAFF-transgenic mice overexpress BAFF and spontaneously develop glomerulonephritis that resembles human IgA nephropathy. We administered IL-33 or PBS to wild-type and BAFF-transgenic mice. After treating Rag1-deficient mice with IL-33, with or without anti-CD90.2 to preferentially deplete ILC2s, we isolated splenocytes, which were adoptively transferred into BAFF-transgenic mice. BAFF-transgenic mice treated with IL-33 developed more severe kidney dysfunction and proteinuria, glomerular sclerosis, tubulointerstitial damage, and glomerular deposition of IgA and C3. Compared with wild-type mice, BAFF-transgenic mice exhibited increases of CD19+ B cells in spleen and kidney and ILC2s in kidney and intestine, which were further increased by administration of IL-33. Administering IL-33 to wild-type mice had no effect on kidney function or histology, nor did it alter the number of ILC2s in spleen, kidney, or intestine. To understand the role of ILC2s, splenocytes were transferred from IL-33-treated Rag1-deficient mice into BAFF-transgenic mice. Glomerulonephritis and IgA deposition were exacerbated by transfer of IL-33-stimulated Rag1-deficient splenocytes, but not by ILC2 (anti-CD90.2)-depleted splenocytes. Wild-type mice infused with IL-33-treated Rag1-deficient splenocytes showed no change in kidney function or ILC2 numbers or distribution. IL-33-expanded ILC2s exacerbated IgA glomerulonephritis in a mouse model. These findings indicate that IL-33 and ILC2s warrant evaluation as possible mediators of human IgA nephropathy.


Exploiting new tools for iron bio-fortification of rice.
Rice is one of the most important staple crops and efficient iron (Fe) adsorption during growth not only improves rice yield, but also enriches this essential micronutrient in rice grains to address Fe deficiency in humans. In this article, we review updates on research into the molecular mechanisms regulating Fe uptake from soil and its transport from roots to shoots to seeds in rice plants. Understanding the regulation and expression of genes involved in Fe homeostasis will benefit the development of variants with enhanced Fe utilization to improve rice output and quality.


Fluorocarbon (FC-43) in augmentation of tissue oxygen and its experimental use for flap survival.
The tissue PO2 level in the skin of rats injected with fluorocarbon (FC-43) was measured by the microelectrode method and by a transcutaneous monitor. It was found that the tissue PO2 in the animals injected with FC-43 was two to three times higher than that of the control group. Neither the administration of FC-43 nor the inhalation of systemic oxygen improved flap survival in the rats.


Monitoring of changes in selenium concentration in goat milk during short-term supplementation of various forms of selenium.
The goal of the experiment was to monitor the changes in the selenium concentration in goat milk during short-term oral supplementation of three different forms of selenium. The experiment involved 24 lactating goats of white shorthaired breed. Group C was the control; group S received selenium in the form of selenium-enriched yeast, group L in the form of lactate, and group B in the form of proteinate. Individual selenium preparations were administered individually orally in 250 microg Se dose per animal for 20 days. After the beginning of selenium supplementation, the selenium concentration in milk during the first 5 days grew gradually in group S. Between days 7 and 20 of Se supplementation, the mean Se concentrations in milk in groups were 12.53 +/- 3.69 microg l(-1) (C), 25.90 +/- 6.30 microg l(-1) (S), 13.14 +/- 3.54 microg l(-1) (L), 11.70 +/- 3.69 microg l(-1) (B). Differences between group S and other groups (C, B, L) were highly significant (p < 0.0001). Based on our results, selenium in the form of lactate and proteinate was excreted into the milk similarly, but selenium in the form of yeast, which contains high amount of selenomethionine, was excreted by milk in the highest amounts.


[Analysis of C2ORF71 gene variant in a Chinese patient with retinitis pigmentosa].
To explore the genetic basis for a Chinese patient with retinitis pigmentosa (RP). Whole exome sequencing (WES) was carried out to screen potential variant in the proband. Candidate variants were determined by taking consideration of clinical phenotype. Sanger sequencing was used to verify the variant in the proband and his parents. The proband was found to harbor compound heterozygous variants of c.8G>A (p.Cys3Tyr) and c.958_959insA (p.Arg320Glnfs*29) in the C2ORF71 gene, which has derived from his father and mother, respectively. Both variants were unreported previously. Based on the ACMG guidelines, they were predicted to be likely pathogenic and pathogenic, respectively. The novel compound heterozygous variants of the C2ORF71 gene probably underlay the pathogenesis of RP in the proband. Above finding has enriched the spectrum of C2ORF71 gene mutations and facilitated genetic counseling for the family.


Components of the fibrinolytic system in the vitreous body in patients with vitreoretinal disorders.
To assess components of the fibrinolytic system in the vitreous humour and serum of patients with vitreoretinal disorders. Forty-three samples of vitreous humour and plasma of 43 patients undergoing pars plana vitrectomy for macular hole, macular pucker, retinal detachment or proliferative vitreoretinopathy were evaluated for their content of plasminogen, a2-antiplasmin, plasminogen-activatorinhibitor 1, plasmin-a2-antiplasmin-complex, tissue plasminogen activator, total protein, albumin, d-dimer. Patient groups were compared with each other using the U-test (Mann and Whitney) for non-parametric testing of two independent samples. The groups of retinal detachment and proliferative vitreoretinopathy had elevated vitreal levels of plasminogenactivator-inhibitor 1 (352.5 and 370.7 ng/mL vs. 1.86 and 56.6 ng/mL, P = non-significant), plasmin-a2-antiplasmincomplex (2416.5 and 1836.2 microg/L vs. 124.2 and 133.4 microg/L, P < 0.001), albumin (0.08 and 0.15 g/dL vs. 0.03 and 0.07 g/dL, P < 0.05) and d-dimer (4.76 and 1.64 microg/mL vs. 0.40 and 0.48 microg/mL, P = non-significant) when compared with patients with macular hole and macular pucker. There are significant differences in the vitreal concentration of components of the fibrinolytic system in patients with vitreoretinal disorders. Breakdown of the blood-retinal barrier and complex disease-specific mechanisms are thought to be responsible for an increase of components of the fibrinolytic system in the vitreous.


The penetration of Fascioloides magna miracidia into the snail host Fossaria bulimoides. A scanning electron microscope study.
Snails of the species Fossaria bulimoides were exposed to miracidia of Fascioloides magna for given periods of time. The course of penetration was followed by means of scanning electron microscopy. Topographical features of the miracidial morphology were studied; the apical organ, epidermal plates and their shedding, and body shapes during penetration. Preliminary data were obtained concerning the nature of the lesions formed by the miracidia. Inferences were drawn concerning the mode and duration of penetration.


Strain energy descriptions of biological swelling. II: Multiple fluid compartment models.
A series of multicompartmental, biphasic elastic tissue models is developed. In its most general form, the models consist of multiple tubular networks, each with an internal spring network. In addition, another spring network occupies the extratubular compartment. Strain energy functions are derived for the models, as well as expressions for the fluid pressures in each compartment arising from volume expansion or swelling. Calculations also show that the distribution of fluid among compartments is a significant determinant of tissue elasticity.


Heavy metal accumulation in lizards living near a phosphate treatment plant: possible transfer of contaminants from aquatic to terrestrial food webs.
We investigated the accumulation of heavy metals in Bosk's fringe-toed lizards (Acanthodactylus boskianus) living in Gabès region (southeastern Tunisia), in relation to habitat, diet, and distance from the Gabès-Ghannouche factory complex of phosphate treatment. More specifically, we compared the concentrations of cadmium, lead, and zinc in the stomach contents and samples of the liver, kidney, and tail from lizards living in four sites corresponding to different combinations of habitat (coastal dunes vs backshore) and distance from the factory complex (<500 vs 20 km). Examination of stomach contents showed that lizards living on the coastal dunes mainly feed on littoral amphipods, while those living in the backshore feed exclusively on terrestrial invertebrates. The concentrations of heavy metals in lizard tissues were overall positively correlated with those in the preys they ingested. Moreover, there was a general tendency towards increased concentrations of cadmium, lead, and zinc in the samples from lizards living on coastal dunes compared to those from the other sites, although some differences still lacked statistical significance. These results suggest that the highest contamination of lizards living on coastal dunes was probably related to the ingestion of contaminated amphipods. Thus, amphipods and Bosk's fringe-toed lizards seem to provide an important link between the marine and terrestrial food webs, with higher concentrations appearing to accumulate from materials released into the sea rather than the terrestrial environment. With regard to metal distribution among tissues, our results were overall in agreement with previous findings in other reptiles. In particular, cadmium was most concentrated in the liver samples, stressing once more the role of the liver as a storage organ of Cd. Moreover, high concentrations of the three assessed metals were found in the kidney samples, showing the role of the kidney as an active site of heavy metal accumulation.


Sensitivity and specificity of FDG PET-CT scan in detecting lymph node metastasis in operable periampullary tumours in correlation with the final histopathology after curative surgery.
18F-Fluorodeoxyglucose-positron emission tomography/computerised tomography (FDG-PET/CT) was investigated for evaluation of periampullary tumours and other gastrointestinal neoplasms. The aim of this study was to evaluate the utility of FDG-PET/CT for detection of lymph node metastasis in periampullary tumours by comparing the preoperative FDG-PET/CT scan finding with postoperative histopathology of lymph nodes. Study was done on 24 patients with diagnosis of periampullary carcinoma either proven or suspected on conventional radiology. Standard uptake value (SUV) were measured for lymph node areas with uptake in FDG-PET/CT and compared with histopathological lymph node status. For detection of lymph node metastasis, FDG-PET/CT with cutoff value SUV max ≥2.0 had a sensitivity of 71.4 % and specificity of 77.8 % and that for SUV max ≥2.5 and 2.8 were 57.1, 42.9 and 77.8, 77.8 %, respectively. The sensitivity and specificity of FDG-PET/CT at each lymph node groups were 72 and 89 % in peripancreatic area, 100 and 93 % in hepatoduodenal area and 100 and 100 % in aortocaval area at SUV max ≥2.0, respectively. At SUV max ≥2.5 the values were 57 and 89 % in peripancreatic area, 100 and 93 % in hepatoduodenal area and 100 and 93 % in aortocaval area. FDG-PET-CT has a possible role in detection of lymph node metastasis in periampullary carcinomas and may be used as a guide for possible lymphadenectomy during surgery and for prognostic purpose.


In vitro activities of fleroxacin, cefetamet, ciprofloxacin, ceftriaxone, trimethoprim-sulfamethoxazole, and amoxicillin-clavulanic acid against rare members of the family Enterobacteriaceae primarily of human (clinical) origin.
Fleroxacin and cefetamet were evaluated in vitro against 38 infrequently encountered species (250 strains) of the family Enterobacteriaceae and compared with four established compounds. For all the strains tested, the fleroxacin MIC was less than or equal to 0.5 mg/liter, and for 98% of strains the cefetamet MIC was less than or equal to 8 mg/liter; even though the two new compounds did not quite reach the activities (on a weight-by-weight basis) of ciprofloxacin and ceftriaxone, respectively, they nonetheless clearly surpassed trimethoprim-sulfamethaxazole and amoxicillin-clavulanic acid. The very potent new oral compounds tested in this study appear to be promising for the treatment of clinically relevant infections due to uncommon species of Enterobacteriaceae.


The U.K. Small Aneurysm Trial: design, methods and progress. The UK Small Aneurysm Trial participants.
To study the outcome of surgery versus ultrasound surveillance for small aortic aneurysms. Multicentre, random allocation, open trial. University and general hospitals across the UK. Patients aged 60-76 years with small asymptomatic abdominal aortic aneurysms (4-5.5 cm in diameter). The mortality rate, costs, usage of health service resources and quality of life of each treatment will be compared. From patients randomised to ultrasound surveillance, the natural history of small aneurysms will be determined. The design, methods and progress of this trial are reported here.


Genotoxic and clastogenic effects of monohaloacetic acid drinking water disinfection by-products in primary human lymphocytes.
The haloacetic acids (HAAs) are the second-most prevalent class of drinking water disinfection by-products formed by chemical disinfectants. Previous studies have determined DNA damage and repair of HAA-induced lesions in mammalian and human cell lines; however, little is known of the genomic DNA and chromosome damage induced by these compounds in primary human cells. The aim of this study was to evaluate the genotoxic and clastogenic effects of the monoHAA disinfection by-products in primary human lymphocytes. All monoHAAs were genotoxic in primary human lymphocytes, the rank order of genotoxicity and cytotoxicity was IAA > BAA >> CAA. After 6 h of repair time, only 50% of the DNA damage (maximum decrease in DNA damage) was repaired compared to the control. This demonstrates that primary human lymphocytes are less efficient in repairing the induced damage by monoHAAs than previous studies with mammalian cell lines. In addition, the monoHAAs induced an increase in the chromosome aberration frequency as a measurement of the clastogenic effect of these compounds. These results coupled with genomic technologies in primary human cells and other mammalian non-cancerous cell lines may lead to the identification of biomarkers that may be employed in feedback loops to aid water chemists and engineers in the overall goal of producing safer drinking water.


Configurations of fatty acyl chains in egg phosphatidylcholine-cholesterol mixed bilayers.
Based on the structural properties of cholesterol and egg phosphatidylcholine, and on the assumption that the van der Waals' type attactive interaction between the steroid nucleus and the fatty acyl chains provides a stabilizing force for the cholesterol-egg phosphatidylcholine complex, some specific orientation and configurations of the fatty acyl chains around the steroid nucleus in the interacting system are proposed in terms of an optimal packing. The proposed model suggests thathe saturated chains are largely facing the flattened (alpha) surface of the steroid nucleus of cholesterol, while the unsaturated chains can interact with both the alpha and beta surfaces of the steroid nucleus. It is also suggested that the angular methyl groups on the beta surface of the steroid nucleus lock the unsaturated fatty acyl chain in a relatively immobile configuration. Experimental evidence which provides support for the proposed stereochemical model is presented.


Manganese(II)-doped carbon dots as effective oxidase mimics for sensitive colorimetric determination of ascorbic acid.
A colorimetric assay is presented for the determination of ascorbic acid (AA). Manganese(II) doped carbon dots (Mn-CDs) were prepared by a convenient hydrothermal route and are shown to possess oxidase-like catalytic ability. They catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by dissolved oxygen to form a blue colored product (oxTMB). AA can reduce blue oxTMB to colorless TMB. The fading of the blue color (measured at 652 nm) can be applied for quantifying AA in the 50 to 2500 nM concentration range and with a 9 nM detection limit. The method was successfully used for the quantitation of AA in real samples. Graphical abstract Schematic representation of a colorimetric assay platform for the sensitive detection of ascorbic acid (AA) in view of inhibitory effect of AA on the 3,3',5,5'-tetramethylbenzidine (TMB) oxidation, benefitting from excellent oxidase-like catalytic activity of manganese(II) doped carbon dots (Mn-CDs).


Risk factors associated with surgical site infection and the development of short-term complications in macaques undergoing indwelling vascular access port placement.
Risk factors associated with surgical site infection (SSI) and the development of short-term complications in macaques undergoing vascular access port (VAP) placement are evaluated in this study. Records from 80 macaques with VAPs were retrospectively reviewed. Logistic regression was used to identify factors associated with short-term post-operative complications. The primary outcome was SSI, which occurred in 21.6% (52.6% in the first 12 months vs. 13% thereafter) of procedures. SSI was associated with major secondary complications including VAP removal (11.4%), wound dehiscence (5.7%), and mechanical catheter occlusion (5.7%). In multivariate modeling, only surgical program progress was a statistically significant predictor of SSI, while animal compliance had a slightly protective effect. Vascular access ports have a moderate risk of complications, provided the surgical program optimizes best practices. Under complex experimental conditions, VAPs represent an important refinement, both improving animals' overall well-being and environment and reducing stress.


STARE-HI--Statement on reporting of evaluation studies in Health Informatics.
Development of guidelines for publication of evaluation studies of Health Informatics applications. An initial list of issues to be addressed in reports on evaluation studies was drafted based on experiences as editors and reviewers of journals in Health Informatics and as authors of systematic reviews of Health Informatics studies, taking into account guidelines for reporting of medical research. This list has been discussed in several rounds by an increasing number of experts in Health Informatics evaluation during conferences and by using e-mail and has been put up for comments on the web. A set of STARE-HI principles to be addressed in papers describing evaluations of Health Informatics interventions is presented. These principles include formulation of title and abstract, of introduction (e.g. scientific background, study objectives), study context (e.g. organizational setting, system details), methods (e.g. study design, outcome measures), results (e.g. study findings, unexpected observations) and discussion and conclusion of an IT evaluation paper. A comprehensive list of principles relevant for properly describing Health Informatics evaluations has been developed. When manuscripts submitted to Health Informatics journals and general medical journals adhere to these aspects, readers will be better positioned to place the studies in a proper context and judge their validity and generalisability. It will also be possible to judge better whether papers will fit in the scope of meta-analyses of Health Informatics interventions. STARE-HI may also be used for study planning and hence positively influence the quality of evaluation studies in Health Informatics. We believe that better publication of both quantitative and qualitative evaluation studies is an important step toward the vision of evidence-based Health Informatics. This study is based on experiences from editors, reviewers, authors of systematic reviews and readers of the scientific literature. The applicability of the principles has not been evaluated in real practice. Only when authors start to use these principles for reporting, shortcomings in the principles will emerge.


Calciphylaxis in a patient with rheumatoid arthritis without renal failure and hyperparathyroidism: the possible role of long-term steroid use and protein S deficiency.
Calciphylaxis is a rare and life-threatening condition of progressive cutaneous necrosis secondary to small and medium-sized vessel calcification that is seen almost exclusively in patients with end-stage renal disease and hyperparathyroidism. We report a case of calciphylaxis that may very well be due to a long-term steroid use in a rheumatoid arthritis patient who had neither end-stage renal disease nor hyperparathyroidism. We also discuss the possible role of protein S deficiency as a contributing factor.


Influence factors of estimation errors for total corneal astigmatism using keratometric astigmatism in patients before cataract surgery.
To evaluate the influence factors of the estimation errors for total corneal astigmatism using keratometric astigmatism in patients preparing for cataract surgery. EYE and ENT Hospital of Fudan University, Shanghai, China. Prospective observational study. Eyes of patients preparing for cataract surgery were measured with Pentacam Scheimpflug imaging device. Keratometric astigmatism was obtained using the anterior corneal surface measurement and the keratometric index (1.3375) while neglecting the posterior corneal surface measurement. The Scheimpflug-measured total corneal astigmatism was derived by vector analysis of the astigmatism on both corneal surfaces. The study comprised 374 eyes of 374 patients 45 to 84 years old. The mean absolute error in magnitude and mean absolute error in angle comparing keratometric astigmatism with Scheimpflug-derived astigmatism was 0.18 ± 0.14 diopter (D) and 7.7 ± 11.0 degrees, respectively. The mean magnitude of the error vector was 0.24 ± 0.14 D. The error in magnitude was significantly larger in eyes with against-the-rule anterior astigmatism, while error in angle was larger in eyes with with-the-rule and oblique anterior astigmatism. Multiple regressions showed that 4 predictors (difference in anterior-posterior astigmatism axis, magnitude of posterior astigmatism, magnitude of keratometric astigmatism, and axial length [AL]) were significantly associated with the absolute error in magnitude. Predictors including the difference in the anterior-posterior astigmatism axis, magnitude of posterior astigmatism, magnitude of keratometric astigmatism, and age were significantly associated with the absolute error in angle and magnitude of the error vector. Neglecting posterior corneal astigmatism yielded significant estimation errors in total corneal astigmatism in patients preparing for cataract surgery. Estimation errors were significantly influenced by the difference in the anterior -posterior astigmatism axis, magnitude of posterior astigmatism, keratometric astigmatism, AL, and age. None of the authors has any conflicts of interest to disclose.


Endothelial function improvement in patients with familial hypercholesterolemia receiving PCSK-9 inhibitors on top of maximally tolerated lipid lowering therapy.
Treatment with protein convertase subtilisin kexin type 9 inhibitors (PCSK-9i) reduced cholesterol levels and cardiovascular events in patients with hypercholesterolemia. We assessed changes in lipid profile, oxidation markers and endothelial function in patients with familial hypercholesterolemia (FH) after a 12-week treatment with a PCSK-9i. Patients with FH starting a treatment with PCSK-9i were included. Total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), lipoprotein(a) (Lp(a)), small dense LDL (assessed by LDL score), 11-dehydro-thromboxane (11-TXB2), 8-isoprostaglandin-2alpha (8-iso-PGF2α), flow-mediated dilation (FMD) and reactive hyperaemia index (RHI) were evaluated before starting PCSK-9i treatment and after a 12-week treatment. Twenty-five subjects were enrolled (52% males, mean age 51.5 years). At the 12-week assessment, we observed a 38% median reduction in TC, 52% in LDL-C, 7% in Lp(a) and 46% in LDL score. In parallel, 11-TXB2 and 8-iso-PGF2α showed a reduction of 18% and 17%, respectively. FMD changed from 4.78% ± 2.27 at baseline to 10.6% ± 5.89 at 12 weeks (p < 0.001), with RHI changing from 2.37 ± 1.23 to 3.76 ± 1.36 (p < 0.001). A multivariate analysis showed that, after adjusting for potential confounders, change in LDL score was an independent predictor of changes in FMD (β = -0.846, p = 0.015) and in 8-iso-PGF2α (β = 0.778, p = 0.012). Small dense LDL reduction (assessed by LDL score) is related to changes in oxidation markers and endothelial function in patients with FH treated with PCSK-9i.


Perceived Unmet Need and Need-Related Distress of People Living With Dementia.
The unmet needs of people living with dementia have been shown to be multidimensional and impact well-being. However, there are a lack of studies examining variability of unmet needs and need-related distress from the person living with dementia's perspective. The current study (n = 12) examined the self-reported unmet needs and need-related distress of people with mild to moderate dementia. Seventy-five percent of participants (n = 9) identified at least one unmet need and 50% (n = 6) reported 10 or more unmet needs. "Finding and Arranging Services" and "Health Information" subscales had the highest reported average unmet needs. The most frequently reported unmet need-item was "getting information about your memory problems?" Participants reported variability in distress for both unmet and met needs. Continued research can provide beneficial information on the relationship between unmet needs, need-related distress, and outcomes of well-being for future interventions.


Humoral immunosuppressant activity of aflatoxin ingestion in rabbits measured by response to Mycobacterium bovis antigens using enzyme-linked immunosorbent assay and serum protein electrophoresis.
A total of 30 New Zealand white rabbits were divided into five equal groups. Animals in groups 1 and 3 were sensitized with bacillus Calmette-Guerin (BCG), and those in groups 2 and 4 with inactivated cells of Mycobacterium bovis (Sensitinogen). Group 1 and 2 rabbits were fed 2 ppm day-1 aflatoxin for 3 months. Group 5 served as control. Serum samples from animals in all groups were subjected to enzyme-linked immunosorbent assays (ELISAs) to determine antibody titre and to protein electrophoresis to determine immunoglobulin levels. The antibody titres and the immunoglobulin levels were significantly decreased in the aflatoxin-treated groups.


A systematic review and meta-analysis to evaluate the diagnostic accuracy of recognition of stroke in the emergency department (ROSIER) scale.
The present study aims to evaluate the performance and the clinical applicability of the Recognition of Stroke in the Emergency Department (ROSIER) scale via systematic review and meta-analysis. Electronic databases of Pubmed and Embase were searched between 1st January 2005 (when ROSIER developed) and 8th May 2020. Studies that evaluated the diagnostic accuracy of the ROSIER scale were included. The sensitivity, specificity, diagnostic odds ratio (DOR), and area under the curve (AUC) were combined using a bivariate mixed-effects model. Fagan nomogram was used to evaluate the clinical applicability of the ROSIER scale. A total of 14 studies incorporating 15 datasets were included in this meta-analysis. The combined sensitivity, specificity, DOR and AUC were 0.88 [95% confidence interval (CI): 0.83-0.91], 0.66 (95% CI: 0.52-0.77), 13.86 (95% CI, 7.67-25.07) and 0.88 (95% CI, 0.85-0.90), respectively. Given the pre-test probability of 60.0%, Fagan nomogram suggested the post-test probability was increased to 79% when the ROSIER was positive. In comparison, it was decreased to 22% when ROSIER was negative. Subgroup analysis showed that the pooled sensitivity of ROSIER in the European population was higher than that in Asia. In contrast, the pooled specificity was not significantly different between them. Moreover, results also suggested the male-to-female ratio ≤ 1.0 subgroup, prehospital setting subgroup, and other trained medical personnel subgroup had significantly higher sensitivity compared with their counterparts. At the same time, no significant differences were found in the pooled specificity between them. ROSIER is a valid scale with high clinical applicability, which has not only good diagnostic accuracy in Europe but also shows excellent performance in Asia. Moreover, the ROSIER scale exhibits good applicability in prehospital settings with other trained medical personnel.


Ataxia with vitamin E deficiency and severe dystonia: report of a case.
Mutation of the gene for alpha-tocopherol transfer protein causes ataxia with isolated vitamin E deficiency, a disorder usually stabilized or improved after vitamin E supplementation. Dystonia has rarely been described in ataxia with isolated vitamin E deficiency (AVED) patients. We present the case of a young boy with AVED, whose neurological and extra-neurological cardinal symptoms of the disease improved after vitamin E supplementation but who progressively developed generalized dystonia.


Correction: Neuronal Cell Fate Specification by the Convergence of Different Spatiotemporal Cues on a Common Terminal Selector Cascade.
[This corrects the article DOI: 10.1371/journal.pbio.1002450.].


Dietary patterns derived by factor analysis are associated with cognitive function among a middle-aged and elder Chinese population.
The aim of this study was to assess the association of empirically derived dietary patterns with cognitive function among a middle-aged and elder Chinese population. This study comprised 1676 Chinese adults ≥45 years of age, who participated in a Health Survey and completed semi-quantitative food frequency questionnaire (FFQ) and cognitive screening test in the city of Linyi.We performed factor analysis using the principal component analysis method to identify the major dietary patterns. Binary logistic regression models were used to estimate odds ratio(OR) and 95% confidence interval(CI) for cognitive impairment according to quartiles of each dietary pattern score. Three dietary patterns were identified, namely traditional Chinese, Western-style and grains-fruits-vegetable patterns. A total of 362 participants (101 men and 261 women) were classified as cognitive impaired. After controlling for potential confounders, participants in the highest quartile of the Western-style pattern had a greater OR for incident cognitive impairment, compared to participants in the lowest quartile. Compared with the lowest quartile of grains-fruits-vegetable pattern, the highest quartile had a lower OR for incident cognitive impairment. Conclusions: Our findings demonstrate that the Western-style pattern is associated with an elevated risk and the grains-fruits-vegetables pattern is associated with a decreased risk of cognitive impairment.


Cellular polyamines condense hyperphosphorylated Tau, triggering Alzheimer's disease.
Many gaps in our understanding of Alzheimer's disease remain despite intense research efforts. One such prominent gap is the mechanism of Tau condensation and fibrillization. One viewpoint is that positively charged Tau is condensed by cytosolic polyanions. However, this hypothesis is likely based on an overestimation of the abundance and stability of cytosolic polyanions and an underestimation of crucial intracellular constituents - the cationic polyamines. Here, we propose an alternative mechanism grounded in cellular biology. We describe extensive molecular dynamics simulations and analysis on physiologically relevant model systems, which suggest that it is not positively charged, unmodified Tau that is condensed by cytosolic polyanions but negatively charged, hyperphosphorylated Tau that is condensed by cytosolic polycations. Our work has broad implications for anti-Alzheimer's research and drug development and the broader field of tauopathies in general, potentially paving the way to future etiologic therapies.


Two abnormal clones in the bone marrow cells of a patient with paroxysmal nocturnal hemoglobinuria.
Paroxysmal nocturnal hemoglobinuria (PNH) is considered to be a clonal disorder, although most investigations have failed to show chromosomal abnormalities. The present patient suffered from PNH and exhibited in bone marrow cells two abnormal clones with 47 chromosomes in addition to cells with 46 chromosomes. One clone showed trisomy 9, a finding previously reported in leukemias and myeloproliferative disorders. Thus, PNH seems to be a clonal myeloproliferative disease.


Long-term survival and late development of bladder cancer in renal transplant patient with progressive multifocal leukoencephalopathy.
We describe a renal transplant recipient in whom progressive multifocal leukoencephalopathy and transitional cell carcinoma of the bladder developed. Despite these potentially fatal sequelae of chronic immunosuppression the patient remains free of recurrent disease.


Arachidonic acid and free fatty acids as second messengers and the role of protein kinase C.
In addition to serving as the precursor to a plethora of eicosanoids and other bioactive molecules, arachidonate may function as a bona fide second messenger. A number of studies have documented the ability of arachidonate to regulate the function of multiple targets in vitro systems. This has been particularly well established and studied with the activation of protein kinase C by arachidonate in a mechanism distinct from activation by diacylglycerol. In cells, arachidonate induces a number of activities, many of which may be independent of further metabolism to eicosanoids; suggesting possible direct action of arachidonate. This review summarizes the current state of knowledge on the possible second messenger function of arachidonate with specific emphasis on the regulation of protein kinase C.


Anxiety during a crisis: emotional effects of screening for neural tube defects.
Anxiety was assessed in prospective mothers undergoing screening for foetal neural tube defects. Anxiety was found to be extreme and only returned to normal levels when a definitely negative amniocentesis result was communicated to the woman. Anxiety was mitigated by social and family support but other life events had little effect on it.


The revision of 8th edition TNM stage criteria is more accurate in prediction postoperative survival for SCLC patients.
The 8th edition TNM lung cancer classification was enacted in January 2017. An important change is the old T2a was revised to new T2a and T2b. Whether a postoperative survival difference exists between the two stages is still undefined. The purpose of this study is to investigate the postoperative outcomes of small cell lung cancer (SCLC) patients and compare the survival difference between the two stages. Data of patients with SCLC who underwent surgery between October 2005 and August 2013 at our institute were reviewed. A total number of 194 patients were enrolled and were reclassified according to the 8th edition TNM stage. Survival was determined by Kaplan-Meier analysis, the log-rank test was used to compare survival differences. Surgical resection included 165 lobectomy or bilobectomy, 19 sublobectomy and 10 pneumonectomy. There were 159 patients received chemotherapy, 35 underwent surgery alone. The 5-year survival rate and median survival time (MST) of whole group was 34.5% and 45 months. For patients who received similar therapy (surgery plus chemotherapy) in N0 stage, the MST of T2a stage was 83 months, significantly longer than 62 months in T2b stage (P = 0.017). In the 8th edition TNM lung cancer stage criteria, the postoperative survival of T2a is superior to T2b for non-metastatic SCLC at N0 stage.


Structure of the O-antigen of Yersinia pseudotuberculosis O:4b.
The structure of the long-chain O-antigen of Yersinia pseudotuberculosis O:4b containing two uncommon deoxy sugars, tyvelose (3,6-dideoxy-d-arabino-hexose, Tyv) and 6-deoxy-d-manno-heptose (D-6dmanHep), was established by 1D and 2D NMR spectroscopy as alpha-Tyvp-(1-->3)-beta-D-6dmanHepp 1-->4 -->3)-alpha-D-Galp-(1-->3)-beta-D-GlcpNac-(1-->.