Inhibition of collagenase from Clostridium histolyticum by phosphoric and phosphonic amides.
Di- and tripeptides with sequences present in collagen that are known to occupy the S1' through S3' subsites at the active site of the collagenase from Clostridium histolyticum do not themselves inhibit this zinc protease. Thus glycylproline, glycylprolylalanine, and their C-terminal amides are not inhibitors. N alpha-Phosphorylglycylproline, N alpha-phosphorylglycyl-L-prolyl-L-alanine, and their C-terminal amides are weak inhibitors with IC50's (concentration causing half-maximal inhibition) of 4.6, 0.8, 3, and 1.5 mM, respectively. Extension of glycyl-L-prolyl-L-alanine to L-leucyl-glycyl-L-prolyl-L-alanine gives a tetrapeptide known to occupy the S1, S1', S2', and S3' subsites of collagenase when present in collagen but that still does not itself inhibit the enzyme. (Isoamylphosphonyl)glycyl-L-prolyl-L-alanine, a peptide containing a tetrahedral phosphorus atom at the position of the amide carbonyl carbon of the L-leucylglycyl amide bond of the parent tetrapeptide, inhibits collagenase with an IC50 of 16 microM, at least 1000-fold more potent than the parent peptide. Substitution of the two-carbon ethyl chain of alanine for the five-carbon isoamyl chain of leucine increases the IC50 to 46 microM. Substitution of the n-decyl chain for the isoamyl chain does not change the IC50. (Isoamylphosphonyl)glycyl-glycyl-L-proline contains a tripeptide that does not occupy the S1' through S3' subsites of collagenase when this peptide is present in collagen and thus has an IC50 of 4.4 mM. (Isoamylphosphonyl)glycyl-L-prolyl-L-alanine may be an analogue of the tetrahedral transition state for the hydrolysis of the natural collagen substrate.(ABSTRACT TRUNCATED AT 250 WORDS)


Ganglioside GM3 inhibits hepatoma cell motility via down-regulating activity of EGFR and PI3K/AKT signaling pathway.
Two related sublines derived from murine ascites hepatoma cell lines Hca-F25, which were selected for their markedly different metastatic potential to lymph nodes, were found to be distinct in their ganglioside patterns. The low metastatic cell line (HcaP) contained a major ganglioside GM3, whereas the high metastatic cell line (HcaF) contained a major ganglioside GM2. Suppression of GM3 by P4 enhanced the mobility and migration of the low metastatic HcaP cells in vitro. Increase in GM3 content in high metastatic HcaF cells by addition of exogenous GM3 inhibited the mobility and migration. These results suggested that the differences in lymphatic metastasis potential between these two cell lines could be attributed to the differences in their ganglioside compositions, and GM3 could suppress the motility and migration of these cells. Further, we investigated the mechanism by which GM3 suppressed the cell mobility and migration. The results showed that suppression of GM3 synthesis by P4 in low metastatic HcaP cells promoted PKB/Akt phosphorylation at Ser473 and Thr308, and phosphorylation of EGFR at the Tyr1173. In contrast, increase in GM3 content in high metastatic HcaF cells by addition of exogenous GM3 into the culture medium suppressed phosphorylation of PKB/Akt and EGFR at the same residues. Taken together, these results suggested that the mechanism of GM3-suppressed cell motility and migration may involve the inhibition of phosphorylation of EGFR and the activity of PI3K/AKT signaling pathway.


[Reflex sympathetic dystrophy syndrome associated with phenobarbital].
Reflex sympathetic dystrophy syndrome (RSDS) is clinically characterized by pain and edema of one or more extremities, trophic skin changes and vasomotor instability. Although the pathogenesis is unknown, it could be caused by an abnormal reflex of the sympathetic nervous system. Different studies haven't yet confirmed the classical division in three clinical phases (warm, of vasomotor instability and cold). Barbiturates are the precipitating event in 10-30% of cases. We describe the clinical features of a patient with RSDS associated with phenobarbital who needed corticosteroid treatment. The Technetium diphosphate bone scan (Tc 99m DPD) is very useful because there is an increased radionuclide uptake in the involved areas during the early phases of the disease and precedes in some weeks the radiologic signs. The Magnetic Resonance Imaging (MRI) may be useful because of the early signs it shows. The patient may develop contractures and atrophy of the involved extremities in spite of the indispensable withdrawal of the drug.


Integration of polarization-multiplexing and phase-shifting in nanometric two dimensional self-mixing measurement.
Integration of phase manipulation and polarization multiplexing was introduced to self-mixing interferometry (SMI) for high-sensitive measurement. Light polarizations were used to increase measuring path number and predict manifold merits for potential applications. Laser source was studied as a microwave-photonic resonator optically-injected by double reflected lights on a two-feedback-factor analytical model. Independent external paths exploited magnesium-oxide doped lithium niobate crystals at perpendicular polarizations to transfer interferometric phases into amplitudes of harmonics. Theoretical resolutions reached angstrom level. By integrating two techniques, this SMI outperformed the conventional single-path SMIs by simultaneous dual-targets measurement on single laser tube with high sensitivity and low speckle noise. In experimental demonstration, by nonlinear filtering method, a custom-made phase-resolved algorithm real-time figured out instantaneous two-dimensional displacements with nanometer resolution. Experimental comparisons to lock-in technique and a commercial Ploytec-5000 laser Doppler velocity meter validated this two-path SMI in micron range without optical cross-talk. Moreover, accuracy subjected to slewing rates of crystals could be flexibly adjusted.


Tecovirimat Resistance in Mpox Patients, United States, 2022-2023.
During the 2022 multinational outbreak of monkeypox virus (MPXV) infection, the antiviral drug tecovirimat (TPOXX; SIGA Technologies, Inc., https://www.siga.com) was deployed in the United States on a large scale for the first time. The MPXV F13L gene homologue encodes the target of tecovirimat, and single amino acid changes in F13 are known to cause resistance to tecovirimat. Genomic sequencing identified 11 mutations previously reported to cause resistance, along with 13 novel mutations. Resistant phenotype was determined using a viral cytopathic effect assay. We tested 124 isolates from 68 patients; 96 isolates from 46 patients were found to have a resistant phenotype. Most resistant isolates were associated with severely immunocompromised mpox patients on multiple courses of tecovirimat treatment, whereas most isolates identified by routine surveillance of patients not treated with tecovirimat remained sensitive. The frequency of resistant viruses remains relatively low (<1%) compared with the total number of patients treated with tecovirimat.


Antibiotic prescriptions in Italian hospitalised children after serial point prevalence surveys (or pointless prevalence surveys): has anything actually changed over the years?
Point prevalence surveys have been used in several studies to provide immediate and easily comparable information about antibiotic use and showed that about one third of hospitalised children had on ongoing antimicrobial prescription during their hospital admission. The aim of this study, as part of the Global Antimicrobial Resistance, Prescribing and Efficacy in Neonates and Children project, is to describe antimicrobial prescriptions among hospitalised children in four tertiary care hospitals in Italy to show if something has changed over the years. Four tertiary care Italian's hospitals joined three Point Prevalence Surveys (PPSs) in three different period of the year. All children under 18 years of age with an ongoing antimicrobial prescription, admitted on the participating wards at 8 o'clock in the morning of the selecting day were enrolled. A total of 1412 patients (475 neonates and 937 children) were admitted in the days of three PPSs. Overall, among the total admitted patients, 565 patients (40%) had an ongoing antimicrobial prescription in the days of the survey A total of 718 antibiotics were administered in the 485 admitted children and 133 in neonates. The most common indications for antibiotic therapy in children was Lower respiratory tract infections (244/718, 34%), while in neonates were prophylaxis for medical problems (35/133, 26.3%), newborn prophylaxis for newborn risk factors (29/133, 21.8%) and prophylaxis for surgical disease (15/133, 11.3%). Based on our results, it appears that nothing has changed since the last PPS and that the quality improved targets, underlyined in previous studies, are always the same. Serial PPSs can be part of AMS strategies but they are not sufficient alone to produce changes in clinical practice.


Nutritional influences on aging of Fischer 344 rats: II. Pathology.
The aim of this study was to explore the effects of nutritional manipulations on the occurrence and progression of age-related pathologic lesions in male Fischer 344 rats. The following nutritional regimens were studied: (a) ad libitum feeding, (b) food restriction initiated at 6 weeks of age, (c) food restriction initiated at 6 months of age, (d) food restriction limited to a period of early life (6 weeks to 6 months of age), (e) protein restriction without caloric restriction. The major age-related lesions observed were chronic nephropathy, cardiomyopathy, and neoplasia. Food restriction initiated at 6 months of age was as effective as food restriction initiated at 6 weeks of age in slowing the progression of chronic nephropathy and cardiomyopathy and in delaying the occurrence of neoplasia. Food restriction limited to early life was much less effective. Protein restriction in the absence of caloric restriction did not delay the occurrence of neoplasia, but it did retard chronic nephropathy and cardiomyopathy, although much less effectively than caloric restriction involving a similar level of protein restriction.


Suicide Intent Scale in the prediction of suicide.
To assess the predictive value of the Suicide Intent Scale in patients with high suicide risk. The secondary aim was to assess if the use of the factors of the Suicide Intent Scale may offer a better predictive value in suicide risk detection. Finally a shorter version of the scale was created after an item analysis. Eighty-one suicide attempters were assessed with the Beck's Suicide Intent Scale (SIS). All patients were followed up for cause of death. Receiver-operating characteristic (ROC) curves and tables were created to establish the optimal cut-off values for SIS and SIS factors to predict suicide. Seven patients committed suicide during a mean follow up of 9.5 years. The major finding was that mean SIS scores distinguished between suicides and survivors. The positive predictive value was 16.7% and the Area Under Curve (AUC) was 0.74. Only the planning subscale reached statistical significance. Four items were used to test a shorter version of the SIS in the suicide prediction. The positive predictive value was 19% and the AUC was 0.82. The Suicide Intent Scale is a valuable tool in clinical suicide risk assessment, a shorter version of the scale may offer a better predictive value.


Characterization and Comparative Evaluation of Milk Protein Variants from Pakistani Dairy Breeds.
The aim of study was to scrutinize the physicochemical and protein profile of milk obtained from local Pakistani breeds of milch animals such as Nilli-Ravi buffalo, Sahiwal cow, Kajli sheep, Beetal goat and Brela camel. Physicochemical analysis unveiled maximum number of total solids and protein found in sheep and minimum in camel. Buffalo milk contains the highest level of fat (7.45%) while camel milk contains minimum (1.94%). Ash was found maximum in buffalo (0.81%) and sheep (0.80%) while minimum in cow's milk (0.71%). Casein and whey proteins were separated by subjecting milk to isoelectric pH and then analyzed through sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The results showed heterogeneity among these species. Different fractions including αS1, αS2, κ-casein, β-casein and β-lactoglobulen (β-Lg) were identified and quantitatively compared in all milk samples. Additionally, this electrophoretic method after examining the number and strength of different protein bands (αS1, αS2, β-CN, α-LAC, BSA, and β-Lg, etc.), was helpful to understand the properties of milk for different processing purposes and could be successfully applied in dairy industry. Results revealed that camel milk was best suitable for producing allergen free milk protein products. Furthermore, based on the variability of milk proteins, it is suggested to clarify the phylogenetic relationships between different cattle breeds and to gather the necessary data to preserve the genetic fund and biodiversity of the local breeds. Thus, the study of milk protein from different breed and species has a wide range of scope in producing diverse protein based dairy products like cheese.


[The prospects for using marine algae].
Different aspects of utilization of the products of marine algae processing are discussed: phycocolloids, sterins, isoprenoids, etc. The natural resources of algae are decreased due to their intensive exploitation. Hence, the development of algal mariculture becomes very urgent.


Time-Restricted Inquiry-Based Learning Promotes Active Student Engagement in Undergraduate Zoology Laboratory.
Organizing a zoology laboratory for an undergraduate course is often a challenge, particularly in a limited-resource setting, due to the vast variety of topics to cover and the limited numbers of preserved specimens and permanent slides. In zoology, the class structure generally takes the form of a lecture demonstration followed by sample exhibition stations. This setting often fails to actively engage the majority of students in exploring the specimens. Here we propose an alternative organization of a zoology class lab format comprised of short guided-inquiry, time-restricted lab stations, and a freely structured follow-up project intended to increase attention and conceptual understanding of the lab topic. The lab is designed in two parts: a 10-minute in-class rotation portion, where small groups of students take turns investigating specimens following an instructor demonstration, and an after-class group assignment. We implemented the strategy for two years, and it is clear that our approach significantly increased students' active engagement in the class. The time-restricted scheme ensures all students participate despite limited resources, while the guided instructions keep the students focused on the topic. Furthermore, the team assignment portion, in particular the media creation aspect, promoted teamwork among group members.


Risk factors governing the development of cerebral vein and dural sinus thrombosis after craniotomy in patients with intracranial tumors.
OBJECTIVE Neurosurgical intervention may increase the risk of developing cerebral vein and dural sinus thrombosis (CVT). The clinical management of CVT in postoperative patients remains unclear. This retrospective study explores the disease occurrence, associated risk factors, and outcomes in patients with tumors who developed CVT after craniotomy. METHODS A retrospective analysis and review of patient records in those who had undergone cranial tumor removal within the authors' neurosurgical department was performed. In so doing, the authors identified a cohort of patients who developed CVT postoperatively. The study included patients who presented to the department between January 2004 and December 2013. RESULTS Of 2286 patients with intracranial lesions who underwent craniotomy, 35 (1.5%) went on to develop CVT. The authors identified the semisitting position (OR 7.55, 95% CI 3.73-15.31, p < 0.001); intraoperative sinus injury (OR 1.5, 95% CI 3.57-15.76, p < 0.001); and known CVT risk factors (OR 7.77, 95% CI 2.28-21.39, p < 0.001) as predictors of CVT development. Of note, 19 patients (54.3%) had good outcomes (modified Rankin Scale Score 0-1), whereas 9 patients (25.7%) had suffered dependency or death (modified Rankin Scale Score 4-6) at last follow-up. Intracerebral hemorrhage (OR 21.27, 95% CI 1.59-285.01, p = 0.02) and delayed delivery of an intermediate dose of low-molecular-weight heparin anticoagulation (OR 24.12, 95% CI 2.08-280.13, p = 0.01) were associated with unfavorable outcomes. CONCLUSIONS Only a minority of patients undergoing craniotomy for tumor removal develop CVT, and the majority of those who do develop CVT recover well. Early administration of an intermediate dose of low-molecular-weight heparin anticoagulation might be considered once CVT is diagnosed.


The Quebec Newborn Twin Study at 21.
This paper is a revised and updated edition of a previous description of the Quebec Newborn Twin Study (QNTS), an ongoing prospective longitudinal follow-up of a birth cohort of twins born between 1995 and 1998 in the greater Montreal area, Québec, Canada. The goal of QNTS is to document individual differences in the cognitive, behavioral, and social-emotional aspects of developmental health across childhood, their early genetic and environmental determinants, as well as their putative role in later social-emotional adjustment, school, health, and occupational outcomes. A total of 662 families of twins were initially assessed when the twins were aged 6 months. These twins and their family were then followed regularly. QNTS now has 16 waves of data collected or planned, including 5 in preschool. Over the last 24 years, a broad range of physiological, cognitive, behavioral, school, and health phenotypes were documented longitudinally through multi-informant and multimethod measurements. QNTS also entails extended and detailed multilevel assessments of proximal (e.g., parenting behaviors, peer relationships) and distal (e.g., family income) features of the child's environment. QNTS children and a subset of their parents have been genotyped, allowing for the computation of a variety of polygenic scores. This detailed longitudinal information makes QNTS uniquely suited for the study of the role of the early years and gene-environment transactions in development.


Mechanical CPR: Who? When? How?
In cardiac arrest, high quality cardiopulmonary resuscitation (CPR) is a key determinant of patient survival. However, delivery of effective chest compressions is often inconsistent, subject to fatigue and practically challenging.Mechanical CPR devices provide an automated way to deliver high-quality CPR. However, large randomised controlled trials of the routine use of mechanical devices in the out-of-hospital setting have found no evidence of improved patient outcome in patients treated with mechanical CPR, compared with manual CPR. The limited data on use during in-hospital cardiac arrest provides preliminary data supporting use of mechanical devices, but this needs to be robustly tested in randomised controlled trials.In situations where high-quality manual chest compressions cannot be safely delivered, the use of a mechanical device may be a reasonable clinical approach. Examples of such situations include ambulance transportation, primary percutaneous coronary intervention, as a bridge to extracorporeal CPR and to facilitate uncontrolled organ donation after circulatory death.The precise time point during a cardiac arrest at which to deploy a mechanical device is uncertain, particularly in patients presenting in a shockable rhythm. The deployment process requires interruptions in chest compression, which may be harmful if the pause is prolonged. It is recommended that use of mechanical devices should occur only in systems where quality assurance mechanisms are in place to monitor and manage pauses associated with deployment.In summary, mechanical CPR devices may provide a useful adjunct to standard treatment in specific situations, but current evidence does not support their routine use.


Electron paramagnetic resonance of the copper in dopamine beta-monooxygenase. Rapid reduction by ascorbate, the steady-state redox level, chelation with EDTA, and reactivation of the apoenzyme by added copper.
The water-soluble form of dopamine beta-monooxygenase from bovine adrenal medulla was studied. Addition of excess CuSO4 to purified enzyme preparations followed by extensive ultrafiltration against copper-free buffer at pH 7.0, gave preparations with about four copper atoms per enzyme tetramer of Mr 290 000. The enzyme-bound copper was shown by the rapid-freeze technique and electron paramagnetic resonance (EPR) to be reduced by ascorbate at a rate which was faster than the overall catalytic rate; about 10% of the copper was oxidized to Cu(II) during steady-state catalysis in the presence of excess ascorbate. These results support an electron-transfer function of the enzyme-bound copper during catalysis and indicate that the reduction by ascorbate is not the rate-limiting step. The enzyme-bound copper was rapidly chelated by EDTA at pH 7.0., and the apoenzyme thus obtained after dialysis revealed no EPR-detectable copper. Addition of CuSO4 to the apoenzyme gave an EPR spectrum similar to that of the native enzyme, and the apoenzyme was fully reactivated by the optimal concentration of CuSO4 in less than 2 s.


Occupational heat illness in Washington State, 1995-2005.
Little information exists describing the incidence of heat-related illness (HRI) among non-military working populations. An analysis of HRI cases utilizing workers' compensation data has not been previously reported. We used both ICD-9 and ANSI Z16.2 codes with subsequent medical record review to identify accepted Washington State Fund workers' compensation claims for HRI over the 11-year time period from 1995-2005. There were 480 Washington workers' compensation claims for HRI during the 11-year study period. NAICS industries with the highest workers' compensation HRI average annual claims incidence rate were Fire Protection 80.8/100,000 FTE, Roofing Construction 59.0/100,000 FTE, and Highway, Bridge and Street Construction 44.8/100,000 FTE. HRI claims were associated with high outdoor ambient temperatures. Medical risk factors for HRI were present in some cases. HRI cases occur in employed populations. HRI rates vary by industry and are comparable to those previously published for the mining industry.


The Modularity of Dysgraphia.
Research regarding dysgraphia, an impairment in writing, is attaining more attention in recent times. The existing studies on dysgraphia draw insights from cognitive, behavioural, neurological, and genetic fields of knowledge. However, these multiple studies on dysgraphia fail to illustrate how these cognitive, behavioural, neurological, and genetic systems interact and intersect in dysgraphia. Therefore, the studies could not offer a comprehensive understanding of dysgraphia. In order to fill this gap, the review attempts to study dysgraphia using the notion of modularity by accommodating insights from cognitive, behavioural, neurological, and genetic aspects of dysgraphia. Such a profound understanding could facilitate an early diagnosis and holistic intervention towards dysgraphia.


Sulfide binding is mediated by zinc ions discovered in the crystal structure of a hydrothermal vent tubeworm hemoglobin.
Key to the remarkable ability of vestimentiferan tubeworms to thrive in the harsh conditions of hydrothermal vents are hemoglobins that permit the sequestration and delivery of hydrogen sulfide and oxygen to chemoautotrophic bacteria. Here, we demonstrate that zinc ions, not free cysteine residues, bind sulfide in vestimentiferan hemoglobins. The crystal structure of the C1 hemoglobin from the hydrothermal vent tubeworm Riftia pachyptila has been determined to 3.15 A and revealed the unexpected presence of 12 tightly bound Zn(2+) ions near the threefold axes of this D(3) symmetric hollow sphere. Chelation experiments on R. pachyptila whole-coelomic fluid and purified hemoglobins reveal a role for Zn(2+) ions in sulfide binding. Free cysteine residues, previously proposed as sulfide-binding sites in vestimentiferan hemoglobins, are found buried in surprisingly hydrophobic pockets below the surface of the R. pachyptila C1 molecule, suggesting that access of these residues to environmental sulfide is restricted. Attempts to reduce the sulfide-binding capacities of R. pachyptila hemoglobins by addition of a thiol inhibitor were also unsuccessful. These findings challenge the currently accepted paradigm of annelid hemoglobin evolution and adaptation to reducing environments.


Impact of physicians' participation in non-interventional post-marketing studies on their prescription habits: A retrospective 2-armed cohort study in Germany.
Non-interventional post-marketing studies (NIPMSs) sponsored by pharmaceutical companies are controversial because, while they are theoretically useful instruments for pharmacovigilance, some authors have hypothesized that they are merely marketing instruments used to influence physicians' prescription behavior. So far, it has not been shown, to our knowledge, whether NIPMSs actually do have an influence on prescription behavior. The objective of this study was therefore to investigate whether physicians' participation in NIPMSs initiated by pharmaceutical companies has an impact on their prescription behavior. In addition, we wanted to analyze whether specific characteristics of NIPMSs have a differing impact on prescription behavior. In a retrospective 2-armed cohort study, the prescription behavior of 6,996 German physicians, of which 2,354 had participated in at least 1 of 24 NIPMSs and 4,642 were controls, was analyzed. Data were acquired between 6 October 2016 and 8 June 2018. Controls were matched by overall prescription volume and number of prescriptions of the drug studied in the NIPMS in the year prior to the NIPMS. Primary outcome was the relative rate of prescriptions of the drug studied in the NIPMS by participating physicians compared to controls during the NIPMS and the following year. Secondary outcomes were the proportion of prescriptions of the studied drug compared to alternative drugs used for the same indication, the revenue generated by these prescriptions, and the association between the marketing characteristics of the NIPMS and prescription habits. Of the 24 NIPMSs, the 2 largest drug groups studied were antineoplastic and immunomodulatory agents (7/24, 29.2%) and agents for the nervous system (4/24, 16.7%). Physicians participating in an NIPMS prescribed more of the studied drug during and in the year after the NIPMS, at a relative rate of 1.08 (95% CI 1.07-1.10; p < 0.001) and 1.07 (95% CI 1.05-1.09); p < 0.001), respectively. Participating physicians were more likely than controls to prescribe one of the studied drugs rather than alternative drugs used for the same indication (odds ratio 1.04; 95% CI 1.03-1.05). None of the marketing characteristics studied were significantly associated with prescription practices. The main limitation was the difficulty in controlling for confounders due to privacy laws, with a resulting lack of information regarding the included physicians, which was mainly addressed by the matching process. Physicians participating in NIPMSs prescribe more of the investigated drug than matching controls. This result calls the alleged non-interventional character of NIPMSs into question and should lead to stricter regulation of NIPMSs.


"All manner of gymnastic evolutions" for science: Dorothea Klumpke (1861-1942) and a life in astronomical research.
In this article, the extraordinary life of the astronomer Dorothea Klumpke (1861-1942) is described in detail for the first time, focussing on the four phases of her career, in which she researched various astronomical questions both as an amateur and as an employee of an observatory and as one half of a couple in science. For this reason, Klumpke's biography provides insights into the cornucopia of research approaches in astronomy at the time, in which professional and amateur astronomers explored the heavens in observatories, on field trips to exotic countries, in their own backyards, or aboard hot air balloons, using telescopes, gazing through the lenses of cameras and spectroscopes, or based on mathematical reasoning. By comparing her life to biographies of other contemporary women, including Klumpke's sisters, among them the famous neurologist Augusta Klumpke-Déjerine, the criteria that women had to fulfill in order to pursue an academic career in the long nineteenth century will be discussed at the same time. In this, particular attention will be paid to factors over which women themselves had no influence, also to show that before the middle of the twentieth century, many stars had to align in order to have such an unusual career as Dorothea Klumpke.


Demonstration of chlamydial inclusions in exfoliated cells.
Microscopic demonstration of chlamydial inclusions within cells offered the first laboratory procedure supporting the clinical diagnosis of chlamydial infection. The cytological test for the detection of chlamydial inclusions in genital tract infection, through not as sensitive as isolation in cell cultures, is still of diagnostic value. The present study discusses the collection of clinical specimens for microscopic examination, the preparation, fixation and staining of slides by Giemsa stain, iodine, and immunofluorescence.


Helicobacter pylori stimulates urokinase plasminogen activator receptor expression and cell invasiveness through reactive oxygen species and NF-kappaB signaling in human gastric carcinoma cells.
The gastric pathogen, helicobacter pylori (H. pylori), has been associated with the progression of gastric cancer. It was previously reported that H. pylori induced urokinase plasminogen activator receptor (uPAR) expression and stimulated cell invasiveness in human gastric cancer AGS cells. However, the precise mechanisms for how H. pylori upregulates uPAR are unclear. This study investigated the underlying signal pathways in H. pylori-induced uPAR in human gastric cancer AGS cells. The intracellular H2O2 content, as determined using H2O2-sensitive probe 2',7'-dichlorodihydrofluorescein, increased after the H. pylori treatment. N-acetyl cysteine (NAC), an antioxidant, prevented the H. pylori-induced production of H2O2 and uPAR expression. In addition, exogenous H2O2 was found to increase uPAR mRNA expression and its promoter activity. Site-directed mutagenesis of the potential NF-kappaB element in the uPAR promoter showed that the redox-sensitive transcription factor NF-kappaB was essential for H. pylori-induced uPAR expression. The expression of vectors encoding a mutated-type NF-kappaB-inducing kinase and I-kappaB, and a specific inhibitor of NF-kappaB (BAY11-7082) decreased the H. pylori-induced uPAR promoter activity. Chromatin immunoprecipitation and the electrophoretic mobility shift assay confirmed that H. pylori increased the DNA binding activity of NF-kappaB. With the aid of NAC and H2O2, it was determined that reactive oxygen species (ROS) is an upstream signaling molecule for activating the NF-kappaB induced by H. pylori. The enhanced AGS cell invasiveness by H. pylori was partially abrogated by an NAC and BAY11-7082 treatment. These results suggest that the ROS and NF-kappaB signaling pathway is important in H. pylori-induced uPAR expression and the increased cell invasiveness of human gastric cancer AGS cells.


Orthogonal invariant sets of the diffusion tensor and the development of a curvilinear set suitable for low-anisotropy tissues.
We develop a curvilinear invariant set of the diffusion tensor which may be applied to Diffusion Tensor Imaging measurements on tissues and porous media. This new set is an alternative to the more common invariants such as fractional anisotropy and the diffusion mode. The alternative invariant set possesses a different structure to the other known invariant sets; the second and third members of the curvilinear set measure the degree of orthotropy and oblateness/prolateness, respectively. The proposed advantage of these invariants is that they may work well in situations of low diffusion anisotropy and isotropy, as is often observed in tissues such as cartilage. We also explore the other orthogonal invariant sets in terms of their geometry in relation to eigenvalue space; a cylindrical set, a spherical set (including fractional anisotropy and the mode), and a log-Euclidean set. These three sets have a common structure. The first invariant measures the magnitude of the diffusion, the second and third invariants capture aspects of the anisotropy; the magnitude of the anisotropy and the shape of the diffusion ellipsoid (the manner in which the anisotropy is realised). We also show a simple method to prove the orthogonality of the invariants within a set.


Determinant and exchange algorithms for observation subset selection.
In many applications involving image reconstruction, signal observation time is limited. This emphasizes the requirement for optimal observation selection algorithms. A selection criterion using the trace of a matrix forms the basis of two existing algorithms, the Sequential Backward Selection and Sequential Forward Selection algorithms. Neither is optimal although both generally perform well. Here we introduce a trace row-exchange criterion to further improve the quality of the selected subset and introduce another observation selection criterion based upon the determinant of a matrix.


Stigma as a fundamental cause of population health inequalities.
Bodies of research pertaining to specific stigmatized statuses have typically developed in separate domains and have focused on single outcomes at 1 level of analysis, thereby obscuring the full significance of stigma as a fundamental driver of population health. Here we provide illustrative evidence on the health consequences of stigma and present a conceptual framework describing the psychological and structural pathways through which stigma influences health. Because of its pervasiveness, its disruption of multiple life domains (e.g., resources, social relationships, and coping behaviors), and its corrosive impact on the health of populations, stigma should be considered alongside the other major organizing concepts for research on social determinants of population health.


Does Acute Otitis Media in the First Month of Life Increase the Risk for Recurrent Otitis?
Acute otitis media (AOM) is a common childhood illness. The aim of this study was to assess whether AOM in the first month of life predicts recurrent AOM (rAOM) in early childhood. The medical records of all neonates with AOM and isolation of bacterial pathogen from middle-ear fluid during 2005-2010 were reviewed. Neonates without AOM admitted during the same period for neonatal fever workup were included as controls. Information regarding rAOM and possible risk factors were collected through a phone interview with the parents. A total of 84 neonates with AOM were enrolled; 25 (30%) had rAOM compared with 8/79 (10%) in the control group. Neonatal AOM increases 4-fold the odds of rAOM later in childhood (odds ratio = 4; 95% CI = 1.44-11.42; P = .008), independent of smoke exposure, numbers of siblings, AOM in siblings, breastfeeding, day care attendance, or use of pacifier. Neonatal AOM is a significant risk factor for rAOM during infancy.


New Horizons in Medical Research.
From the above it is seen that the U. S. Public Health Service Research Grants program represents a sincere and continuing effort to supply Federal funds for the support of necessary additional research in the fields of medical and related sciences without interposing any degree of government restriction, control, supervision, or regimentation. The program is a scientific one, scientific guidance of which lies wholly in the hands of scientists.


The arabidopsis DNA polymerase δ has a role in the deposition of transcriptionally active epigenetic marks, development and flowering.
DNA replication is a key process in living organisms. DNA polymerase α (Polα) initiates strand synthesis, which is performed by Polε and Polδ in leading and lagging strands, respectively. Whereas loss of DNA polymerase activity is incompatible with life, viable mutants of Polα and Polε were isolated, allowing the identification of their functions beyond DNA replication. In contrast, no viable mutants in the Polδ polymerase-domain were reported in multicellular organisms. Here we identify such a mutant which is also thermosensitive. Mutant plants were unable to complete development at 28°C, looked normal at 18°C, but displayed increased expression of DNA replication-stress marker genes, homologous recombination and lysine 4 histone 3 trimethylation at the SEPALLATA3 (SEP3) locus at 24°C, which correlated with ectopic expression of SEP3. Surprisingly, high expression of SEP3 in vascular tissue promoted FLOWERING LOCUS T (FT) expression, forming a positive feedback loop with SEP3 and leading to early flowering and curly leaves phenotypes. These results strongly suggest that the DNA polymerase δ is required for the proper establishment of transcriptionally active epigenetic marks and that its failure might affect development by affecting the epigenetic control of master genes.


Colon cancer presenting as Streptococcus gallolyticus infective endocarditis.
A 58-year-old Chinese man presented with a three-week history of fever. He had a background history of rheumatic heart disease, hypertension, and thalassaemia. He was found to have infective endocarditis of the aortic valve due to Streptococcus gallolyticus. During the hospital stay, he developed a few episodes of haematochaezia and was subsequently found to have colonic carcinomain- situ. He completed appropriate antibiotic treatment for his infective endocarditis and underwent a left hemicolectomy with primary anastomosis. The association between Streptococcus gallolyticus infective endocarditis and colonic neoplasm is well documented. This case report stresses the importance of performing routine colonoscopy to look for colonic neoplastic change in patients diagnosed to have Streptococcus gallolyticus infective endocarditis. The early diagnosis of the colonic neoplasm has enabled our patient to have a curative surgery without compromising his quality of life.


A novel Escherichia coli-derived mutation detected with the Big Blue cII mutant selectable assay.
Transgenic mouse mutation detection systems allow investigation of the origins and mechanisms of mutation associated with exogenous and endogenous mutagen exposures in individual tissues and cell types. In the past, selection assays for transgenic mutants have been contaminated with nonmurine-derived mutations and assay validation is critical to ensure murine in vivo origins of mutations. This is critical in studies of spontaneous mutations and extrapolation to endogenous mammalian genes. Herein, we provide one measure of the contribution of Escherichia coli (E. coli)-derived mutations to the Big Blue(R) cII transgene mutant selection assay. We report the first direct evidence of an E. coli-derived cII mutation identified among mutations recovered in the cII selective assay. An E. coli transposable 5 (Tn5) element IS50R inverted repeat (1,534 bp) was identified at base pair 414 in the cII transgene and the insertion generated a 9 bp target site duplication typical of this type of transposition. The bacterial transposition occurred only once in the assay of 25 x 10(6) plaque forming units and sequencing of 1,177 cII mutants. The observed frequency of this type of mutation is 4 x 10(-8) in retrieved lambda phage and 8.5 x 10(-4) in harvested cII mutants and thus a very rare occurrence in typical analyses of spontaneous in vivo mutations. Given that the frequency of transposition is equal to, or an order of magnitude higher, than the frequency of point mutations in E. coli, this article provides excellent validation for the murine origins of mutations detected using the cII mutant selection assay.


Phosphoproteomic Changes Induced by Cell-Derived Matrix and Their Effect on Tumor Cell Migration and Cytoskeleton Remodeling.
Increased fibrotic extracellular matrix (ECM) deposition promotes tumor invasion, which is the first step of the metastatic cascade. Yet, the underlying mechanisms are poorly understood as conventional studies of tumor cell migration are often performed in 2D cultures lacking the compositional and structural complexity of native ECM. Moreover, these studies frequently focus on select candidate pathways potentially overlooking other relevant changes in cell signaling. Here, we combine a cell-derived matrix (CDM) model with phosphotyrosine phosphoproteomic analysis to investigate tumor cell migration on fibrotic ECM relative to standard tissue culture plastic (TCP). Our results suggest that tumor cells cultured on CDMs migrate faster and in a more directional manner than their counterparts on TCP. These changes in migration correlate with decreased cell spreading and increased cell elongation. While the formation of phosphorylated focal adhesion kinase (pFAK)+ adhesion complexes did not vary between TCP and CDMs, time-dependent phosphoproteomic analysis identified that the SRC family kinase LYN may be differentially regulated. Pharmacological inhibition of LYN decreased tumor cell migration and cytoskeletal rearrangement on CDMs and also on TCP, suggesting that LYN regulates tumor cell migration on CDMs in combination with other mechanisms. These data highlight how the combination of physicochemically complex in vitro systems with phosphoproteomics can help identify signaling mechanisms by which the fibrotic ECM regulates tumor cell migration.


The effect of the spread of the new COVID-19 on the psychological and social adaptation of families of persons with disabilities in the Kingdom of Saudi Arabia.
Helping families of persons with disabilities in times of the spread of COVID-19 is a major step to ensure their social and psychological adaptation. This call comes from the fact that from a health perspective, persons with disabilities have less immunity and are more vulnerable to deceases and infections. Many families of persons with disabilities have also expressed their concerns regarding the safety of their disabled member/s. Therefore, this study aimed to identify the impact of the new COVID-19 on the psychological and social adjustment of families of persons with disabilities in the Kingdom of Saudi Arabia. The sample of the study was chosen randomly from the Asir region. The sample consisted of any member of the families of persons with disabilities (parents and siblings). Participants responded to a questionnaire, which was sent electronically via WhatsApp to the families of persons with disabilities. The questionnaire gathered demographic information about the disabled member from the family (age and level of disability). The questionnaire underwent statistical treatment to extract data for analysis. The results showed less ability for families of persons with disabilities to adapt psychologically and socially to the spread of COVID-19. The findings revealed the existence of fundamental differences in the levels of psychological and social adaptation among these families due to the level of disability of their member and due to the variable of age, and in favor of the age group of 11-15 years. The study recommended providing safe methods of communication with families of persons with disabilities to reduce the feeling of psychological isolation and loneliness as much as possible during the quarantine or curfew periods. This study explored the levels of psychological and social adaptation of families of persons with disabilities in the time of COVID-19. The spread of the virus created a crisis in different counties and millions of people were infected. Persons with disabilities are among the categories affected most by the infection of the virus due to their weak immunity and their health problems. Therefore, response plans need to take measures that cover the needs of persons with disabilities and their families.


Animal leptospirosis in Malaya: 1. Methods, zoogeographical background, and broad analysis of results.
In recent years leptospirosis has been shown to be an important cause of human febrile illness in Malaya. Studies were therefore undertaken to determine its animal reservoirs and the factors influencing spread of infection from them to man and domestic animals. This paper presents the board picture obtained. A wide range of animal species were trapped in forest localities, ricefield areas, areas of scrub and cultivation and in several towns and villages. The maintenance hosts of leptospirosis in Malaya appear to be mainly or entirely rats, although evidence of infection has been found throughout the animal kingdom. Some rat species have characteristics which suggest that they are better maintenance hosts than others. Evidence was found of practically every serogroup of leptospires infecting animals in Malaya. Altogether 104 strains were isolated and identified, and 155 animals were found to have serological evidence of infection. Of 1763 rodents examined, 194 had evidence of infection, and 41 of 1083 other animals. A serum survey of domestic animals showed the highest incidence of antibodies to be in goats and the lowest in oxen.


Animal transportation networks.
Many group-living animals construct transportation networks of trails, galleries and burrows by modifying the environment to facilitate faster, safer or more efficient movement. Animal transportation networks can have direct influences on the fitness of individuals, whereas the shape and structure of transportation networks can influence community dynamics by facilitating contacts between different individuals and species. In this review, we discuss three key areas in the study of animal transportation networks: the topological properties of networks, network morphogenesis and growth, and the behaviour of network users. We present a brief primer on elements of network theory, and then discuss the different ways in which animal groups deal with the fundamental trade-off between the competing network properties of travel efficiency, robustness and infrastructure cost. We consider how the behaviour of network users can impact network efficiency, and call for studies that integrate both network topology and user behaviour. We finish with a prospectus for future research.


Pharmacokinetics and efficacy of pemetrexed in patients with brain or leptomeningeal metastases.
Brain metastases (BM) and leptomeningeal metastases (LM) are devastating neurologic complications. Pemetrexed is a multi-targeted anti-folate agent approved for treatment of nonsquamous non-small cell lung cancer but has anti-tumor activity in other solid tumors. We performed two trials using pemetrexed in patients with BM and LM to assess CSF penetration and anti-tumor activity. Patients were treated with intravenous pemetrexed at doses of 500 (n = 3), 750 (n = 3), 900 (n = 12) or 1,050 mg/m(2) (n = 3) every 3 weeks. Neuro-imaging was done every 6 weeks. Matched CSF and plasma samples were obtained serially from three patients with Ommaya reservoirs; the remaining patients had a single paired collection. Twenty-one patients (15 women and six men) with median age of 50 years and median KPS of 90 were treated. Primary tumors included breast (13), lung (4), colorectal (1), endometrial (1), esophageal (1) and pinealoblastoma (1). Nine patients had prior whole brain RT and median number of prior chemotherapies was two including prior methotrexate in four patients. Median pemetrexed doses administered was three (range 1-14). Responses included one partial response, ten stable disease and ten progressive disease. Median time to progression and survival was 2.7 and 7.3 months; PFS six was 22 %. No major toxicities were seen. Pemetrexed distributed from the plasma to the CSF within 1-4 h with the resulting CSF concentrations < 5 % of plasma. Pemetrexed was tolerated in solid tumor patients with CNS metastases. Limited anti-tumor activity was seen, which might have been due to low CSF concentrations, although some patients displayed prolonged benefit.


Lon protease inactivation, or translocation of the lon gene, potentiate bacterial evolution to antibiotic resistance.
Previous work demonstrated that selection for Escherichia coli mutants with low antibiotic resistance frequently resulted in co-selection of lon mutations and that lon(-) mutants evolved higher-level resistance faster than a lon(+) strain. Here we show that lon mutation causes a very low multidrug resistance by inducing the AcrAB-TolC pump via stabilization of the acrAB transcriptional activators MarA and SoxS, which are substrates of the Lon protease. Fast evolution of lon(-) mutants towards higher resistance involves selection of frequent next-step mutations consisting of large duplications including acrAB and the mutated lon gene. Resistance results from the combined effects of acrAB duplication and lon mutation increasing dosage of efflux pump. In contrast, when acrAB duplication occurs as the first step mutation, increased Lon activity caused by lon(+) co-duplication mitigates the effect of acrAB duplication on resistance, and faster evolution towards higher resistance is not observed. As predicted, when the functional lon gene is relocated far from acrAB to prevent their co-duplication, first-step acrAB duplication confers higher resistance, which then allows selection of frequent next-step mutations and results in faster evolution towards higher resistance. Our results demonstrate how order of appearance of mutations and gene location can influence the rate of resistance evolution.


Herd health monitoring in dairy farms - discover metabolic diseases. An overview.
The transition period, 3 weeks before and 3 weeks after calving, is inevitably accompanied by a negative energy balance, which sometimes causes metabolic disturbances, such as ketosis. Subclinical ketosis (SCK) is defined as an increase in the β-hydroxybutyrate (BHB) concentration to ≥ 1.2 mmol/l in the blood. According to a recent study, a value of ≥ 0.7 mmol/l of non-esterified fatty acids (NEFA) in the blood indicates the potential development of the poor metabolic adaption syndrome (PMAS). With a herd prevalence of 21 %, and an incidence of approximately 40 % within the first 2 weeks after calving, SCK is a relevant herd health problem. The milk yield decreases in the first 2 weeks postpartum by 3-5.3 kg/d for each ketotic cow, and the total milk reduction through the whole lactation period of 305 days averages 112 kg (SD 89 kg). Although the cow does not display any clinical signs of ketosis at this stage, the risk of developing associated production diseases like retained placenta, metritis, displaced abomasum, lameness and clinical ketosis increases and the expected performance in terms of milk production will decrease. The herd health status deteriorates and the risk for early culling increases. Another impact factor is the financial aspect, which includes costs for early death, reduced milk production, reproduction losses, and associated production diseases. In the literature, the calculated costs per SCK case vary between $ 78 and $ 289. The gold standard diagnostic test for SCK is the photometric measurement of BHB in blood. This method is accurate, but results are delayed due to the required laboratory analysis. There are also some rapid cow-side tests, i. e. urine or milk strip tests available to identify ketotic cows. The common disadvantage of these methods is that they are not suitable for herd health monitoring because of the need to collect samples from each cow manually and the high rates of false negative results. However, Fourier transform infrared spectroscopy is suitable for herd health monitoring. It is already being used for the analysis of milk composition. This inexpensive, rapid and simple technique has a specificity of 83.8 % and a sensitivity of 82.4 %. Therefore, FTIR is an early and easy method for detecting ketotic cows, that could help reduce financial and performance losses associated with ketosis.


The diagnosis and management of ovarian hyperstimulation syndrome.
To review the clinical aspects of ovarian hyperstimulation syndrome and provide recommendations on its diagnosis and clinical management. These guidelines will assist in the early recognition and management of ovarian hyperstimulation. Early recognition and prompt systematic supportive care will help avert poor outcomes. Medline, Embase, and the Cochrane database were searched for relevant articles, using the key words "ovarian hyperstimulation syndrome" and "gonadotropins," and guidelines created by other professional societies were reviewed. The quality of evidence was rated using the criteria described in the Report of the Canadian Task Force on Preventive Health Care. Recommendations for practice were ranked according to the method described in that report (Table 1). 1. Once the diagnosis of ovarian hyperstimulation syndrome is made, disease severity should be classified as mild, moderate, severe, or critical. (III-B) 2. The physician prescribing gonadotropins should inform each woman of her personal risk for ovarian hyperstimulation syndrome. (III-A) 3. In areas where patients do not have ready access to physicians familiar with the diagnosis and management of ovarian hyperstimulation syndrome, the physician prescribing gonadotropins should ensure that women are made aware that they should contact a physician or a member of the team within the hospital unit who has relevant experience, should the need arise. (III-B) 4. Outpatient management is recommended for women with mild and moderate ovarian hyperstimulation syndrome. If outpatient management for more severe ovarian hyperstimulation syndrome is to be undertaken, the physician should ensure that the woman is capable of adhering to clinical instructions and that there is a system in place to assess her status every 1 to 2 days. (III-A) 5. Paracentesis should be performed in admitted patients with tense ascites to alleviate their discomfort. (II-2B) 6. Outpatient culdocentesis should be considered for the prevention of disease progression in moderate or severe ovarian hyperstimulation syndrome. (II-2B) 7. Women with severe and critical ovarian hyperstimulation syndrome should be admitted to hospital for intravenous hydration and observation. (III-A) 8. Intravenous hydration should be initiated with a crystalloid solution to prevent hemoconcentration and provide adequate end-organ perfusion. If end-organ perfusion is not maintained with a crystalloid solution, an alternate colloid solution should be administered. (II-2B) 9. Pain relief in admitted patients should be managed with acetaminophen and/or opioid analgesics. (III-B) Non-steroidal anti-inflammatory drugs with antiplatelet properties should not be used. (III-B) 10. Women with severe ovarian hyperstimulation syndrome should be considered for treatment with prophylactic doses of anticoagulants. (II-2B) 11. Critical ovarian hyperstimulation syndrome should be managed by a multidisciplinary team, according to the end organ affected. (III-C).


Multiple functional ECG signal is processing for wearable applications of long-term cardiac monitoring.
In this paper, an integrated electrocardiogram (ECG) signal-processing scheme is proposed. Using a systematic wavelet transform algorithm, this signal-processing scheme can realize multiple functions in real time, including baseline-drift removal, noise suppression, QRS detection, heart beat rate prediction and classification, and clean ECG reconstruction. Utilizing the novel low-cost hardware architecture, the proposed ECG signal-processing scheme is implemented in application-specific integrated circuits with 0.18 μ m CMOS technology. This ECG signal-processor chip achieves low area and power consumptions, and is highly suitable for wearable applications of long-term cardiac monitoring.


A novel hypothesis regarding the evolutionary origins of the immunoglobulin fold.
The recent cloning of two cDNAs (Clone 1 and Clone 5) that encode novel hypothetical proteins, combining an N-terminal Ig kappa-like domain with features that occur in microfibril-associated glycoproteins (MAGPs) and fibrinogen, raises the question of whether the Ig fold may have originated in association with functions that may be more primitive than soluble immunity. Pairwise alignments were performed to compare similarities of fibrinogen-beta, Clone 1 and an Ig kappa sequence. Clone 1 had two regions in its Ig domain with > 50% similarity to fibrinogen, while Ig kappa was virtually non-homologous to fibrinogen. This result suggests that Clone 1 is closer to their common ancestor. A neighbour-joining tree was computed, and it supported this interpretation. Three-dimensional modelling of the most highly conserved sequence revealed two antiparallel beta strands connected by a helix. These observations suggest that the ancestral gene for the immunoglobulin superfamily may have originated as a primitive sandwich-like fold, possibly used in matrix/cell communications.


The biosecurity of on-farm mortality composting.
Composting is a natural biological decomposition process that takes place under aerobic and thermophilic conditions. It can be used for the day-to-day management of mortalities on farms and for carcass disposal in emergency animal disease (EAD) outbreaks. In mortality composting, carcasses are placed in piles or bins together with supplemental carbon sources such as sawdust, litter, straw or wood shavings. Composting is particularly suitable for broiler-farm mortalities and litter. In the case of emergency disease outbreaks, composting can be conducted either inside or outside the poultry house following killing. Composting has been successfully employed for emergency disposal of carcasses in a few cases in North America, but research is lacking on the biosecurity of the process. Composting is a well-established pathogen reduction technology, but process management and heterogenous pile conditions pose particular challenges for validating the microbiological safety of mortality composting. This paper reviews the available information on the biosecurity of mortality composting, identifies potential sources of risk, and highlights emerging research needs. Reports to date of the use of composting in EAD outbreaks are also discussed.


Does an oral care protocol reduce VAP in patients with a tracheostomy?
Several studies have demonstrated that oral care with chlorhexidine gluconate (CHG) 0.12% solution reduces the incidence of ventilator-associated pneumonia (VAP) in mechanically ventilated patients with endotracheal tubes in the ICU. Minimal evidence shows the effectiveness of any oral care protocols in preventing VAP in mechanically ventilated patients with tracheostomies in a step-down or progressive care unit (PCU). To determine the effectiveness of an oral care protocol in reducing the VAP rate in mechanically ventilated patients with tracheostomies in the PCU. A 12-month prospective study was conducted on 75 mechanically ventilated patients who had tracheostomies. The oral care protocol consisted of tooth brushing with toothpaste and applying CHG 0.12% solution every 12 hours. At the conclusion of the study, the VAP rate in the study population was compared with the National Health and Safety Network (NHSN) report for 2009 benchmark of 1.5 per 1,000 ventilator days. After the oral care protocol was implemented in the PCU, the VAP rate was 1.1 per 1,000 ventilator days over 12 months, compared with the NHSN report for 2009 of 1.5 per 1,000 ventilator days. Tooth brushing with toothpaste and applying CHG 0.12% solution may be an effective oral care protocol to reduce the VAP rate in patients in PCUs with tracheostomies who are being mechanically ventilated.


Seasonality of endemic COVID-19.
Successive waves of infection by SARS-CoV-2 have left little doubt that this virus will transition to an endemic disease. Foreknowledge of when to expect seasonal surges is crucial for healthcare and public health decision-making. However, the future seasonality of COVID-19 remains uncertain. Evaluating its seasonality is complicated due to the limited years of SARS-CoV-2 circulation, pandemic dynamics, and varied interventions. In this study, we project the expected endemic seasonality by employing a phylogenetic ancestral and descendant state approach that leverages long-term data on the incidence of circulating HCoV coronaviruses. Our projections indicate asynchronous surges of SARS-CoV-2 across different locations in the northern hemisphere, occurring between October and January in New York and between January and March in Yamagata, Japan. This knowledge of spatiotemporal surges leads to medical preparedness and enables the implementation of targeted public health interventions to mitigate COVID-19 transmission.IMPORTANCEThe seasonality of COVID-19 is important for effective healthcare and public health decision-making. Previous waves of SARS-CoV-2 infections have indicated that the virus will likely persist as an endemic pathogen with distinct surges. However, the timing and patterns of potentially seasonal surges remain uncertain, rendering effective public health policies uninformed and in danger of poorly anticipating opportunities for intervention, such as well-timed booster vaccination drives. Applying an evolutionary approach to long-term data on closely related circulating coronaviruses, our research provides projections of seasonal surges that should be expected at major temperate population centers. These projections enable local public health efforts that are tailored to expected surges at specific locales or regions. This knowledge is crucial for enhancing medical preparedness and facilitating the implementation of targeted public health interventions.


Plant resistance to mechanical stress: evidence of an avoidance-tolerance trade-off.
External mechanical forces resulting from the pressure exerted by wind or water movement are a major stress factor for plants and may cause regular disturbances in many ecosystems. A plant's ability to resist these forces relies either on minimizing the forces encountered by the plant (avoidance strategy), or on maximizing its resistance to breakage (tolerance strategy). We investigated plant resistance strategies using aquatic vegetation as a model, and examined whether avoidance and tolerance are negatively correlated. We tested the avoidance-tolerance correlation across 28 species using a phylogenetically corrected analysis, after construction of a molecular phylogeny for the species considered. Different species demonstrated contrasting avoidance and tolerance and we demonstrated a significant negative relationship between the two strategies, which suggests an avoidance-tolerance trade-off. Negative relationships may result from costs that each strategy incurs or from constraints imposed by physical laws on plant tissues. The existence of such a trade-off has important ecological and evolutionary consequences. It would lead to constraints on the evolution and variation of both strategies, possibly limiting their evolution and may constrain many morphological, anatomical and architectural traits that underlie avoidance and tolerance.


In vivo kinetics of [F-18]MEFWAY: a comparison with [C-11]WAY100635 and [F-18]MPPF in the nonhuman primate.
[F-18]Mefway was developed to provide an F-18 labeled positron emission tomography (PET) neuroligand with high affinity for the serotonin 5-HT(1A) receptor to improve the in vivo assessment of the 5-HT(1A) system. The goal of this work was to compare the in vivo kinetics of [F-18]mefway, [F-18]MPPF, and [C-11]WAY100635 in the rhesus monkey. Each of four monkeys were given bolus injections of [F-18]mefway, [C-11]WAY100635, and [F-18]MPPF and scans were acquired with a microPET P4 scanner. Arterial blood was sampled to assay parent compound throughout the time course of the PET experiment. Time activity curves were extracted in the high 5-HT(1A) binding areas of the anterior cingulate cortex (ACG), mesial temporal cortex, raphe nuclei, and insula cortex. Time activity curves were also extracted in the cerebellum, which was used as a reference region. The in vivo kinetics of the radiotracers were compared based on the nondisplaceable distribution volume (V(ND) ) and binding potential (BP(ND) ). At 30 min, the fraction of radioactivity in the plasma due to parent compound was 19%, 28%, and 29% and cleared from the arterial plasma at rates of 0.0031, 0.0078, and 0.0069 (min⁻¹) ([F-18]mefway, [F-18]MPPF, [C-11]WAY100635). The BP(ND) in the brain regions were mesial temporal cortex: 7.4 ± 0.6, 3.1 ± 0.4, 7.0 ± 1.2, ACG: 7.2 ± 1.2, 2.1 ± 0.2, 7.9 ± 1.2; raphe nuclei: 3.7 ± 0.6, 1.3 ± 0.3, 3.3 ± 0.7; and insula cortex: 4.2 ± 0.6, 1.2 ± 0.1, 4.7 ± 1.0 for [F-18]mefway, [F-18]MPPF, and [C-11]WAY100635 respectively. In the rhesus monkey, [F-18]mefway has similar in vivo kinetics to [C-11]WAY100635 and yields greater than 2-fold higher BP(ND) than [F-18]MPPF. These properties make [F-18]mefway a promising radiotracer for 5-HT(1A) assay, providing higher counting statistics and a greater dynamic range in BP(ND).


Restrictive Spirometry Pattern Is Associated With Increased Arterial Stiffness in Men and Women.
There is growing evidence that pulmonary function impairment is related to cardiovascular events and death. Some studies have shown that the level of FVC is negatively related to arterial stiffness, but most studies were confined to men, and none of them examined the association of the presence of restrictive spirometry pattern with arterial stiffness. Therefore, this study aimed to investigate the association of restrictive spirometry pattern with arterial stiffness by sex. This study recruited 2,961 subjects after excluding those with (1) obstructive lung disease, as defined by history and pulmonary function test; (2) history of asthma, lung cancer, tuberculosis, coronary heart disease, stroke, or any pulmonary structural deformities; and (3) medications influencing BP, plasma glucose, lipid profile, and pulmonary function test. Restrictive spirometry pattern was diagnosed as an FVC < 80% of the predicted value and an FEV1/FVC ratio ≥ 70%. Increased arterial stiffness was defined as right brachial ankle pulse wave velocity (baPWV) ≥ 1,400 cm/s. In both men and women, FVC was negatively associated with the baPWV level. Restrictive spirometry pattern was positively associated with increased arterial stiffness in both men and women (men: OR, 2.16; 95% CI, 1.33-3.50; women: OR, 1.95; 95% CI, 1.02-3.72) after adjustment for other clinical variables. Both restrictive spirometry pattern and reduced FVC were associated with a higher risk of arterial stiffness, not only in men but also in women. Clinically, assessment of arterial stiffness might be considered in individuals with restrictive spirometry pattern.


[Hyperhomocysteinemia: risk factor for premature atheromatosis].
Hyperhomocysteinemia is an independent risk factor for atherosclerosis and cardiovascular disease. The cause of hyperhomocysteinemia is either an inborn metabolic defect or acquired. Main causes are either a defective homocysteine remethylation (thermolability of the enzyme 5,10-methylenetetrahydrofolate reductase) or nutritional deficiencies of B vitamins especially folic acid. The relative risk for myocardial infarction has been found of 3,1 in case of hyperhomocysteinemia. It is considered that a 5 microM/l homocysteine increment elevates vascular risk by as much as cholesterol increases of 20 mg/dl. B vitamins supplements are potentially useful.


Comparison of in vivo efficacy of different ocular lubricants in dry eye animal models.
To compare the efficacy of three types of ocular lubricants in protecting corneal epithelial cells in dry eye animal models. Ocular lubricants containing 0.1% or 0.3% sodium hyaluronate (SH), carboxymethylcellulose (CMC), or hydroxypropyl methylcellulose (HPMC) were tested. First, ocular lubricant containing 0.002% fluorescein was dropped onto the rabbit corneas. The fluorescein intensity as an index of retention was measured. Second, a rabbit dry eye model was made by holding the eye open with a speculum, and 50 μL of each ocular lubricant was dropped onto the cornea. After 3 hours, the corneas were stained with 1% methylene blue (MB), and the absorbance of MB was measured. Third, a rat dry eye model was treated with the ocular lubricants for 4 weeks, and the corneal fluorescein staining was scored. Eyes treated with physiological saline were used as controls. Finally, immunohistochemistry was used to analyze occludin, an epithelial barrier protein, in cultured human corneal epithelial cells pretreated with ocular lubricants and desiccated for 20 or 60 minutes. Our results showed that 0.3% SH had a significantly longer retention time than the other lubricants (all P < 0.01). The absorbance of MB was significantly lower in the 0.3% SH group. The corneas of rats exposed to 0.3% SH had significantly lower fluorescein staining scores. A significantly higher number of occludin-positive cells were found after exposure to 0.3% SH than other lubricants. Ocular lubricant containing 0.3% SH would be preferable to treat patients with dry eye syndrome.


Brain Regional Blood Flow and Working Memory Performance Predict Change in Blood Pressure Over 2 Years.
Hypertension is a presumptive risk factor for premature cognitive decline. However, lowering blood pressure (BP) does not uniformly reverse cognitive decline, suggesting that high BP per se may not cause cognitive decline. We hypothesized that essential hypertension has initial effects on the brain that, over time, manifest as cognitive dysfunction in conjunction with both brain vascular abnormalities and systemic BP elevation. Accordingly, we tested whether neuropsychological function and brain blood flow responses to cognitive challenges among prehypertensive individuals would predict subsequent progression of BP. Midlife adults (n=154; mean age, 49; 45% men) with prehypertensive BP underwent neuropsychological testing and assessment of regional cerebral blood flow (rCBF) response to cognitive challenges. Neuropsychological performance measures were derived for verbal and logical memory (memory), executive function, working memory, mental efficiency, and attention. A pseudo-continuous arterial spin labeling magnetic resonance imaging sequence compared rCBF responses with control and active phases of cognitive challenges. Brain areas previously associated with BP were grouped into composites for frontoparietal, frontostriatal, and insular-subcortical rCBF areas. Multiple regression models tested whether BP after 2 years was predicted by initial BP, initial neuropsychological scores, and initial rCBF responses to cognitive challenge. The neuropsychological composite of working memory (standardized beta, -0.276; se=0.116; P=0.02) and the frontostriatal rCBF response to cognitive challenge (standardized beta, 0.234; se=0.108; P=0.03) significantly predicted follow-up BP. Initial BP failed to significantly predict subsequent cognitive performance or rCBF. Changes in brain function may precede or co-occur with progression of BP toward hypertensive levels in midlife.


Central and peripheral nervous system complications.
Symptomatic neuropathy is a common manifestation of diabetes mellitus, and sensory, motor, or autonomic symptoms occur in approximately 10% of all diabetic patients. Animal models may be useful to study the metabolic and electrophysiologic abnormalities peculiar to diabetic neuropathy. Genetic animal models, including the Chinese hamster, ob/ob mouse, db/db mouse, BB-wistar rat, and SSDR rat or chemically induced or nutritional models of diabetes mellitus provide the potential to use animals to study human neuropathy; however, to date, few characteristics of human diabetic neuropathy have been clearly demonstrated in any of these animal models. Better characterization of the neuropathy of existing animal models with emphasis on evaluation over long periods of time is recommended. These studies should include a cross-disciplinary approach using biochemical, electrophysiologic and morphologic techniques. Specific future approaches to study diabetic neuropathy using chemical models is outlined in this chapter.


Human Papillomavirus Infection in Penile Cancer: Multidimensional Mechanisms and Vaccine Strategies.
Penile cancer (PC) is a rare male malignant tumor, with early lymph node metastasis and poor prognosis. Human papillomavirus (HPV) plays a key role in the carcinogenesis of PC. This review aims to summarize the association between HPV infection and PC in terms of virus-host genome integration patterns (the disrupted regions in the HPV and PC genome), genetic alterations, and epigenetic regulation (methylation and microRNA modification) occurring in HPV and PC DNA, as well as tumor immune microenvironment reprogramming. In addition, the potential of HPV vaccination strategies for PC prevention and treatment is discussed. Understanding of the HPV-related multidimensional mechanisms and the application of HPV vaccines will promote rational and novel management of PC.


Genetic variation of G4P[6] rotaviruses: evidence for novel strains circulating between the hospital and community.
One hundred forty-six fecal specimens collected between 2007 and 2008 from infants with acute gastroenteritis were screened for rotavirus by ELISA with VP6-specific antibody. One hundred twenty-three of the samples (84.2%) were confirmed to be positive for group A rotavirus (community-acquired, n = 90 [73.2%] and nosocomial, n = 33 [26.8%]), and were typed subsequently using RT-PCR and sequence analysis methods. Determination of G- and P-type combinations showed that G4P[6] (78.9%) was the most common strain, followed by G3P[8] (7.3%), G1P[8] (6.5%), G2P[4] (0.8%), G2P[6] (0.8%), G1P[6] (0.8%), and G9P[8] (0.8%) strains. Of the 97 G4P[6] strains, 62 (63.8%) were responsible for community-acquired cases and 35 (36.1%) were hospital-acquired cases. Phylogenetic analysis of the VP7 gene from the G4P[6] strains revealed that both the community-acquired and nosocomial strains were segregated to the human rotaviruses circulating world-wide, including the prototype vaccinal strain, ST3, which constituted a novel sublineage in lineage 1. Owing to the recent emergence of G4P[6] rotaviruses within the hospital, as well as in the community, the findings from this study are important since they provide new information concerning the community and nosocomial spread of rotaviruses.


A Decision Support System for Pathology Test Result Reviews in an Emergency Department to Support Patient Safety and Increase Efficiency.
The review of pathology test results for missed diagnoses in Emergency Departments is time-consuming, laborious, and can be inaccurate. An automated solution, with text mining and clinical terminology semantic capabilities, was developed to provide clinical decision support. The system focused on the review of microbiology test results that contained information on culture strains and their antibiotic sensitivities, both of which can have a significant impact on ongoing patient safety and clinical care. The system was highly effective at identifying abnormal test results, reducing the number of test results for review by 92%. Furthermore, the system reconciled antibiotic sensitivities with documented antibiotic prescriptions in discharge summaries to identify patient follow-ups with a 91% F-measure - allowing for the accurate prioritization of cases for review. The system dramatically increases accuracy, efficiency, and supports patient safety by ensuring important diagnoses are recognized and correct antibiotics are prescribed.


Unidirectional flux ratio for potassium ions in depolarized frog skeletal muscle.
Small bundles of frog skeletal muscle fibers were loaded with 305 mM K+ and 120 mM Cl-, and 42K+ tracer efflux and influx were measured as a function of external K+ concentration ([K+]o) at a resting potential of -2 mV. As [K+]o was lowered from 305 mM, efflux decreased along a markedly sigmoidal curve, reaching a constant nonzero value at low [K+]o. Influx varied linearly with [K+]o at low [K+]o and more steeply at higher [K+]o. The ratio of influx to efflux was described by the equation: influx/efflux = exp[-n(V - VK)F/RT] with n = 2 at high [K+]o, but the ratio approached this equation with n = 1 at low [K+]o. Efflux did not depend on [K+]o when the membrane potential was raised to +36 mV, whereas at low [K+]o decreasing the membrane potential to -19 mV further activated the efflux. The results are discussed in terms of an inwardly rectifying potassium channel with two or more activating sites within the membrane that bind K+ and are accessible from the external solution.


Applying a Dynamical Systems Model and Network Theory to Major Depressive Disorder.
Mental disorders like major depressive disorder can be modeled as complex dynamical systems. In this study we investigate the dynamic behavior of individuals to see whether or not we can expect a transition to another mood state. We introduce a mean field model to a binomial process, where we reduce a dynamic multidimensional system (stochastic cellular automaton) to a one-dimensional system to analyse the dynamics. Using maximum likelihood estimation, we can estimate the parameter of interest which, in combination with a bifurcation diagram, reflects the expectancy that someone has to transition to another mood state. After numerically illustrating the proposed method with simulated data, we apply this method to two empirical examples, where we show its use in a clinical sample consisting of patients diagnosed with major depressive disorder, and a general population sample. Results showed that the majority of the clinical sample was categorized as having an expectancy for a transition, while the majority of the general population sample did not have this expectancy. We conclude that the mean field model has great potential in assessing the expectancy for a transition between mood states. With some extensions it could, in the future, aid clinical therapists in the treatment of depressed patients.


Resveratrol-Encapsulated Glutathione-Modified Robust Mesoporous Silica Nanoparticles as an Antibacterial and Antibiofilm Coating Agent for Medical Devices.
The emergence of various lethal bacterial infections and their adherence to medical devices are major public health concerns. The increased bacterial exposure and titer are accompanied by the inappropriate use of antibiotics that sometimes lead to antibiotic resistance, and therefore, a drug-free antibacterial approach is required. Several nanoparticles (NPs) have been developed as antibacterial and antibiofilm coating agents, which can overcome different drug resistance mechanisms by inhibiting the important processes related to bacterial virulence potential. However, developing safe and biocompatible nanomaterials (NMs) for these applications has remained a major challenge due to their poorly understood mechanism of action. In this work, biogenic silica NPs were modified with glutathione (GSH) to form GSH@SNP (∼80 ± 15 nm) for targeting the bacterial cell surface and biofilm. GSH@SNP was loaded with resveratrol to obtain Res_GSH@SNP (∼124 ± 15 nm) that enhances the antibacterial activity of the NPs against Staphylococcus aureus and Escherichia coli by ∼51 and ∼49%, respectively, compared to GSH@SNP. Res_GSH@SNP is responsible for binding to the bacterial cell surface receptors that interrupt the cell membrane potential, leading to reactive oxygen species (ROS) generation, membrane disruption, and DNA damage and eventually resulting in antibacterial activity. Moreover, the antibiofilm activity of Res_GSH@SNP has been found to result from the interaction of the NPs with the abundant carbohydrates present on the biofilm surface. To check the practical utility of Res_GSH@SNP, these were further evaluated as an antibacterial and antibiofilm coating agent for urinary catheters and were found to be effective even after multiple washes. Res_GSH@SNP has been found to exhibit ∼80 ± 1.4% cytocompatibility toward fibroblast NIH-3T3 cells. Overall, this study is expected to pave the way for the development of biocompatible NP-based coating agents for medical devices.


Tricuspid valve regurgitation following temporary or permanent endocardial lead insertion, and the impact of cardiac resynchronization therapy.
While some studies indicate that permanent pacemaker implantation is associated with development of tricuspid regurgitation (TR), other studies indicate no association.Little is known about the impact of temporary lead insertion during ablation procedures, or whether therapy (CRT) prevents TR post-device implantation. We hypothesized that permanent, but nottemporary endocardial leads, are associated with development of TR, and that CRT would prevent (physiologic) TR. We performed a retrospective study of consecutive patients who underwent first device or radiofrequency catheter ablation over a 12-month period at a single, tertiary academic center who underwent pre- and post-procedure echocardiography. In the 89 patients in the device group, the degree of TR significantly increased ≥ 1 grade post-permanent lead implantation: 9 had less TR, 46 were unchanged, and 34 had more TR(p=0.005). TR increased in the 62 patients who underwent device implantation without CRT (p=0.005), but did not increase in the 27 patients with CRT (p=0.47). In the 66 patients in the ablation group, there was no significant change in TR post-ablation: 8 had less TR, 48 were unchanged, and 10 had more TR (p=0.31). Permanent endocardial lead implantation was associated with an increase in TR; however, patients who underwent device implantation with CRT did not have an increase in TR.Temporary lead insertion during ablation was not associated with changes in the degree of TR. A large, prospective study is needed to accurately define the incidence and exact mechanisms of permanent endocardial lead-related TR.


Inferring a district-based hierarchical structure of social contacts from census data.
Researchers have recently paid attention to social contact patterns among individuals due to their useful applications in such areas as epidemic evaluation and control, public health decisions, chronic disease research and social network research. Although some studies have estimated social contact patterns from social networks and surveys, few have considered how to infer the hierarchical structure of social contacts directly from census data. In this paper, we focus on inferring an individual's social contact patterns from detailed census data, and generate various types of social contact patterns such as hierarchical-district-structure-based, cross-district and age-district-based patterns. We evaluate newly generated contact patterns derived from detailed 2011 Hong Kong census data by incorporating them into a model and simulation of the 2009 Hong Kong H1N1 epidemic. We then compare the newly generated social contact patterns with the mixing patterns that are often used in the literature, and draw the following conclusions. First, the generation of social contact patterns based on a hierarchical district structure allows for simulations at different district levels. Second, the newly generated social contact patterns reflect individuals social contacts. Third, the newly generated social contact patterns improve the accuracy of the SEIR-based epidemic model.


Bioresorbable Materials on the Rise: From Electronic Components and Physical Sensors to In Vivo Monitoring Systems.
Over the last decade, scientists have dreamed about the development of a bioresorbable technology that exploits a new class of electrical, optical, and sensing components able to operate in physiological conditions for a prescribed time and then disappear, being made of materials that fully dissolve in vivo with biologically benign byproducts upon external stimulation. The final goal is to engineer these components into transient implantable systems that directly interact with organs, tissues, and biofluids in real-time, retrieve clinical parameters, and provide therapeutic actions tailored to the disease and patient clinical evolution, and then biodegrade without the need for device-retrieving surgery that may cause tissue lesion or infection. Here, the major results achieved in bioresorbable technology are critically reviewed, with a bottom-up approach that starts from a rational analysis of dissolution chemistry and kinetics, and biocompatibility of bioresorbable materials, then moves to in vivo performance and stability of electrical and optical bioresorbable components, and eventually focuses on the integration of such components into bioresorbable systems for clinically relevant applications. Finally, the technology readiness levels (TRLs) achieved for the different bioresorbable devices and systems are assessed, hence the open challenges are analyzed and future directions for advancing the technology are envisaged.


Impact of Space Research on Science and Technology: Space techniques have provided new tools for science and applications.
Science and technology are themselves a part of our modern culture. They contribute to the arts, furnish new media for human expression, provide better and new musical instruments and improved acoustics for enriched enjoyment of them, and give the artist new materials and techniques for his work. They enrich the humanities in furnishing man with a broader basis for understanding himself, his potentialities, and his place in nature. The historian has a better approach to his subject because of scientific methodology. The archeologist has a surer grasp on temporal relationships because of the tools that physics and chemistry give him for dating materials and identifying their sources. Slowly, an entire world absorbs the increased understanding of man and nature that science generates, and reflects that understanding in its literature, its social, political, and economic institutions, and its application to human daily living. Today, much of the world's population is aware of the world as a body of a solar system in a galaxy among millions of other galaxies. These people can conceive of man in a historical and cosmological perspective that did not exist in centuries past. As this understanding and these concepts spread to more and more of the world's people, it may be hoped that this common bond of understanding will not only give increased motivation to solve peaceably the problems that beset the world, but will also provide increased means for doing so.


Role of collagenase in mediating in vitro alveolar epithelial wound repair.
Type II pneumocytes are essential for repair of the injured alveolar epithelium. The effect of two MMP collagenases, MMP-1 and MMP-13 on alveolar epithelial repair was studied in vitro. The A549 alveolar epithelial cell line and primary rat alveolar epithelial cell cultures were used. Cell adhesion and cell migration were measured with and without exogenous MMP-1. Wound healing of a cell monolayer of rat alveolar epithelial cell after a mechanical injury was evaluated by time lapse video analysis. Cell adhesion on type I collagen, as well as cytoskeleton stiffness, was decreased in the presence of exogenous collagenases. A similar decrease was observed when cell adhesion was tested on collagen that was first incubated with MMP-1 (versus control on intact collagen). Cell migration on type I collagen was promoted by collagenases. Wound healing of an alveolar epithelial cell monolayer was enhanced in the presence of exogenous collagenases. Our results suggest that collagenases could modulate the repair process by decreasing cell adhesion and cell stiffness, and by increasing cell migration on type I collagen. Collagen degradation could modify cell adhesion sites and collagen degradation peptides could induce alveolar type II pneumocyte migration. New insights regarding alveolar epithelial cell migration are particularly relevant to investigate early events during alveolar epithelial repair following lung injury.


Use and endocytosis of iron-containing proteins by Entamoeba histolytica trophozoites.
Iron is essential for nearly all organisms; in mammals, it is part of proteins such as haemoglobin, and it is captured by transferrin and lactoferrin. Transferrin is present in serum, and lactoferrin is secreted by the mucosa and by neutrophils at infection sites, as a host iron-withholding response, sequestering iron away from invading microorganisms. Additionally, all cells contain ferritin, which sequesters iron when its intracellular levels are increased, detoxifying and preventing damage. Liver ferritin contains 50% of iron corporal reserves. During evolution, pathogens have evolved diverse strategies to obtain iron from their hosts in order to survive. The protozoan Entamoeba histolytica invades the intestinal mucosa, causing dysentery, and the trophozoites often travel to the liver producing hepatic abscesses; thus, intestine and liver proteins could be important iron supplies for E. histolytica. We found that E. histolytica trophozoites can grow in both ferrous and ferric iron, and that they can use haemoglobin, holo-transferrin, holo-lactoferrin, and ferritin as in vitro iron sources. These proteins supported the amoeba growth throughout consecutive passages, similarly to ferric citrate. By confocal microscopy and immunoblotting, iron-binding proteins were observed specifically bound to the amoeba surface, and they were endocytosed, trafficked through the endosomal/lysosomal route, and degraded by neutral and acidic cysteine-proteases. Transferrin and ferritin were mainly internalized through clathrin-coated vesicles, and holo-lactoferrin was mainly internalized by caveola-like structures. In contrast, apo-lactoferrin bound to membrane lipids and cholesterol, inducing cell death. The results suggest that in vivo trophozoites secrete products that can destroy enterocytes, erythrocytes, and hepatocytes, releasing transferrin, haemoglobin, ferritin, and other iron-containing proteins, which, together with lactoferrin derived from neutrophils and acinar cells, could be used as abundant iron supplies by amoebas.


Anti-obesity effects of α-cyclodextrin-stabilized 4-methylthio-3-butenyl isothiocyanate from daikon (Raphanus sativus var. longipinnatus) in mice.
4-Methylthio-3-butenyl isothiocyanate (MTBI) is a pungent bioactive constituent found in daikon. However, MTBI is immediately hydrolyzed to 3-hydroxy-methylene-2-thioxopyrrolidine in grated daikon. In this study, we evaluated whether MTBI in grated daikon complexed with α-cyclodextrin (αCD) has anti-obesity effects in mice. C57BL/6J mice were fed a normal diet (normal group), high-fat diet (HFD, control group), HFD with αCD (αCD group), or HFD with MTBI-αCD (MTBI-αCD group) for 16 weeks. The results showed that the final body weight, epididymal white adipose tissue weight, and plasma triglyceride and total cholesterol levels were significantly lower in the MTBI-αCD group than in the control group. The cell size in epididymal adipose tissue was significantly smaller and the accumulation of lipids in the liver was significantly lower in the MTBI-αCD group than in the control group. Furthermore, real-time polymerase chain reaction showed that the mRNA expression level of tumor necrosis factor-alpha was suppressed in the MTBI-αCD group. We also observed low superoxide dismutase activity in the MTBI-αCD group, possibly because MTBI-αCD has the potential to resist HFD-induced oxidative injury. In conclusion, MTBI-αCD exerted anti-inflammation and antioxidant effects to suppress lipid accumulation in epididymal adipose tissue and the liver. These effects then prevented HFD-induced obesity in mice.


Juvenile nasopharyngeal angiofibroma.
The juvenile nasopharyngeal angiofibroma, a locally invasive, non-metastasizing tumour of male adolescence, occurs sporadically throughout the world. Its histiogenesis remains uncertain and its management controversial. These facets of a troublesome and dangerous conditions are discussed and the experiences from handling 9 such tumours seen at Groote Schuur Hospital over a 4-year period (1976-1979) are presented. Surgical excision, using a wide field exposure and pre-operative systemic oestrogen medication, is recommended as the treatment of choice.


FADS1 genetic variability interacts with dietary α-linolenic acid intake to affect serum non-HDL-cholesterol concentrations in European adolescents.
Two rate-limiting enzymes in PUFA biosynthesis, Δ5- and Δ6-desaturases, are encoded by the FADS1 and FADS2 genes, respectively. Genetic variants in the FADS1-FADS2 gene cluster are associated with changes in plasma concentrations of PUFA, HDL- and LDL-cholesterol, and TG. However, little is known about whether dietary PUFA intake modulates these associations, especially in adolescents. We assessed whether dietary linoleic acid (LA) or α-linolenic acid (ALA) modulate the association between the FADS1 rs174546 polymorphism and concentrations of PUFA, other lipids, and lipoproteins in adolescents. Dietary intakes of LA and ALA, FADS1 rs174546 genotypes, PUFA levels in serum phospholipids, and serum concentrations of TG, cholesterol, and lipoproteins were determined in 573 European adolescents from the HELENA study. The sample was stratified according to the median dietary LA (≤9.4 and >9.4 g/d) and ALA (≤1.4 and >1.4 g/d) intakes. The associations between FADS1 rs174546 and concentrations of PUFA, TG, cholesterol, and lipoproteins were not affected by dietary LA intake (all P-interaction > 0.05). Similarly, the association between the FADS1 rs174546 polymorphism and serum phospholipid concentrations of ALA or EPA was not modified by dietary ALA intake (all P-interaction > 0.05). In contrast, the rs174546 minor allele was associated with lower total cholesterol concentrations (P = 0.01 under the dominant model) and non-HDL-cholesterol concentrations (P = 0.02 under the dominant model) in the high-ALA-intake group but not in the low-ALA-intake group (P-interaction = 0.01). These results suggest that dietary ALA intake modulates the association between FADS1 rs174546 and serum total and non-HDL-cholesterol concentrations at a young age.


Dense packing in the monodisperse hard-sphere system: a numerical study.
We report a numerical study of the close packing of monodisperse hard spheres. The close packings of hard spheres are produced by the Lubachesky-Stillinger (LS) compression algorithm and span the range from the disordered states to the ordered states. We provide quantitative evidence for the claim that the density and structural order of the arrested close packing can be determined by the compression rate, i.e., with slower rates producing denser and more ordered structures. Through deeply analyzing the structure of the resulting arrested close packings, a transition region has been identified in the plane of density and reciprocal compression rate, in between what have been historically thought of as amorphous and crystalline packings. We also find clear system size dependences in studying the structural properties of the packings from the disordered ones to the ordered ones. These detailed investigations, on the structure of the arrested close packings, may provide a link between the glassy states and the crystalline states in the hard spheres.


Transcending the challenge of evolving resistance mechanisms in Pseudomonas aeruginosa through β-lactam-enhancer-mechanism-based cefepime/zidebactam.
Multi-drug resistant (MDR) Pseudomonas aeruginosa harbor a complex array of β-lactamases and non-enzymatic resistance mechanisms. In this study, the activity of a β-lactam/β-lactam-enhancer, cefepime/zidebactam, and novel β-lactam/β-lactamase inhibitor combinations was determined against an MDR phenotype-enriched, challenge panel of P. aeruginosa (n = 108). Isolates were multi-clonal as they belonged to at least 29 distinct sequence types (STs) and harbored metallo-β-lactamases, serine β-lactamases, penicillin binding protein (PBP) mutations, and other non-enzymatic resistance mechanisms. Ceftazidime/avibactam, ceftolozane/tazobactam, imipenem/relebactam, and cefepime/taniborbactam demonstrated MIC90s of >128 mg/L, while cefepime/zidebactam MIC90 was 16 mg/L. In a neutropenic-murine lung infection model, a cefepime/zidebactam human epithelial-lining fluid-simulated regimen achieved or exceeded a translational end point of 1-log10 kill for the isolates with elevated cefepime/zidebactam MICs (16-32 mg/L), harboring VIM-2 or KPC-2 and alterations in PBP2 and PBP3. In the same model, to assess the impact of zidebactam on the pharmacodynamic (PD) requirement of cefepime, dose-fractionation studies were undertaken employing cefepime-susceptible P. aeruginosa isolates. Administered alone, cefepime required 47%-68% fT >MIC for stasis to ~1 log10 kill effect, while cefepime in the presence of zidebactam required just 8%-16% for >2 log10 kill effect, thus, providing the pharmacokinetic/PD basis for in vivo efficacy of cefepime/zidebactam against isolates with MICs up to 32 mg/L. Unlike β-lactam/β-lactamase inhibitors, β-lactam enhancer mechanism-based cefepime/zidebactam shows a potential to transcend the challenge of ever-evolving resistance mechanisms by targeting multiple PBPs and overcoming diverse β-lactamases including carbapenemases in P. aeruginosa.IMPORTANCECompared to other genera of Gram-negative pathogens, Pseudomonas is adept in acquiring complex non-enzymatic and enzymatic resistance mechanisms thus remaining a challenge to even novel antibiotics including recently developed β-lactam and β-lactamase inhibitor combinations. This study shows that the novel β-lactam enhancer approach enables cefepime/zidebactam to overcome both non-enzymatic and enzymatic resistance mechanisms associated with a challenging panel of P. aeruginosa. This study highlights that the β-lactam enhancer mechanism is a promising alternative to the conventional β-lactam/β-lactamase inhibitor approach in combating ever-evolving MDR P. aeruginosa.