Three-dimensional structure of the ternary complex between ribonuclease T1, guanosine 3',5'-bisphosphate and inorganic phosphate at 0.19 nm resolution.
The ternary complex formed between RNase T1, guanosine 3',5'-bisphosphate (3',5'-pGp) and Pi crystallizes in the cubic space group I23 with a = 8.706(1) nm. In a previous publication [Lenz, A., Heinemann, U., Maslowska, M. & Saenger, W. (1991) Acta Crystallogr. B47, 521-527], the structure of the complex (in which Pi was not located) was described at a resolution of 0.32 nm. This is now extended to 0.19 nm with newly grown, larger crystals. Refinement with restrained least-squares converged at R = 17.8% for 8027 reflections with [Fo[ > or = 1 sigma ([Fo[); the final model comprises 120 water molecules. 3',5'-pGp is bound to RNase T1 in the anti form, with guanine in the specific recognition site; the 3'-phosphate protrudes into the solvent, and the 5'-phosphate hydrogen bonds with Lys41 O and Asn43 N4. A tetrahedral anion assigned as Pi occupies the catalytic site and hydrogen bonds to the side chains of Tyr38, Glu58, Arg77 and His92. The overall polypeptide fold of RNase T1 in the cubic space group does not differ significantly from that in the orthorhombic space group P2(1)2(1)2(1) except for changes < or = 0.2 nm in loop regions 69-72 and 95-98.


Effects of prenatal ethanol exposure on hypothalamic-pituitary-adrenal function across the estrous cycle.
Rats prenatally exposed to ethanol (E) typically show increased hypothalamic-pituitary-adrenal (HPA) responses to stressors in adulthood. Importantly, prenatal ethanol may differentially alter stress responsiveness in male and female offspring, suggesting a role for the gonadal hormones in mediating the effects of ethanol on HPA activity. We investigated the role of ethanol-induced changes in hypothalamic-pituitary-gonadal (HPG) activity in the differential HPA regulation observed in E compared to control females across the estrous cycle. Peripheral hormones and changes in central neuropeptide mRNA levels were measured across the estrous cycle in adult female offspring from E, pair-fed (PF) and ad libitum-fed control (C) dams. Ethanol females showed normal estrous cyclicity (vaginal smears) but delayed sexual maturation (vaginal opening). Both HPG and HPA activity were differentially altered in E (and in some cases, PF) compared to control females as a function of estrous cycle stage. In relation to HPG activity, E and PF females had higher basal and stress estradiol (E(2)) levels in proestrus compared to other phases of the cycle, and decreased GnRH mRNA levels compared to C females in diestrus. Further, E females had greater variation in LH than PF and C females across the cycle, and in proestrus, only E females showed a significant LH increase following stress. In relation to HPA activity, both basal and stress CORT levels and overall ACTH levels were greater in E than in C females in proestrus. Furthermore, AVP mRNA levels were increased overall in E compared to PF and C females. These data demonstrate ethanol-induced changes in both HPG and HPA activity that are estrous phase-specific, and support the possibility that changes in HPA activity in E females may reflect differential sensitivity to ovarian steroids. E females appear to have an increased HPA sensitivity to E(2), and a possible shift toward AVP regulation of HPA activity. That PF were similar to E females on some measures suggests that nutritional effects of diet or food restriction played a role in mediating at least some of the changes observed.


Eating green. Consumers' willingness to adopt ecological food consumption behaviors.
Food consumption is associated with various environmental impacts, and consumers' food choices therefore represent important environmental decisions. In a large-scale survey, we examined consumers' beliefs about ecological food consumption and their willingness to adopt such behaviors. Additionally, we investigated in more detail how different motives and food-related attitudes influenced consumers' willingness to reduce meat consumption and to buy seasonal fruits and vegetables. We found consumers believed avoiding excessive packaging had the strongest impact on the environment, whereas they rated purchasing organic food and reducing meat consumption as least environmentally beneficial. Similarly, respondents appeared to be most unwilling to reduce meat consumption and purchase organic food. Taste and environmental motives influenced consumers' willingness to eat seasonal fruits and vegetables, whereas preparedness to reduce meat consumption was influenced by health and ethical motives. Women and respondents who preferred natural foods were more willing to adopt ecological food consumption patterns.


Two eyes in action.
Do relative binocular disparities guide our movements in depth? In order to find out we asked subjects to move a 'cursor' to a target within a simulated horizontal plane at eye height. They did so by moving a computer mouse. We determined how quickly subjects responded to the target jumping in depth. We found that it took subjects about 200 ms to respond to changes in binocular disparity. Subjects responded just as quickly if the cursor was temporarily only visible to one eye near the time that the target jumped in depth, and less vigorously, though just as quickly, if the cursor jumped rather than the target, so the fastest binocular responses cannot be based directly on the relative retinal disparity between the target and the cursor. Subjects reacted faster to changes in the target's height in the visual field than to changes in binocular disparity, but did not react faster to changes in image size. These results suggest that binocular vision mainly improves people's everyday movements by giving them a better sense of the distances of relevant objects, rather than by relative retinal disparities being used to directly guide the movement. We propose that relative disparities only guide parts of very slow movements that require extreme precision.


Comparison of clinical test and questionnaires for the evaluation of upper limb prosthetic use in children.
The aim of the present study was to find out if there was a correlation between an observational clinical test and a questionnaire for the evaluation of upper limb prosthetic use in children and to determine which one was better and easier to use in clinical practice. Twenty children who were patients of the children's prosthetic clinic at the Rehabilitation Institute in Ljubljana, Slovenia, and had a functional prosthesis were included in the study. The age appropriate subtest of the University of New Brunswick Test of Prosthetic Function (the UNB test) was assessed by an occupational therapist. Parents completed either the Child Amputee Prosthetics Project-Functional Status Inventory for Preschool children (CAPP-FSIP) or the Child Amputee Prosthetics Project-Functional Status Inventory (CAPP-FSI), depending on which was appropriate for the child's age. Information was limited to the upper extremity items. We found a significant correlation between UNB spontaneity and skill score (r = 0.956, p = 0.000) and also between the parental CAPP score and UNB test (UNB spontaneity--CAPP activities r = 0.634, p = 0.003; UNB spontaneity--CAPP prosthetic use r = 0.542, p = 0.014, UNB skill--CAPP activities r = 0.559, p = 0.010, UNB skill--CAPP prosthetic use r = 0.597, p = 0.005). We concluded that both instruments can be used for assessing upper limb prosthetic use in children but neither is an optimal choice.


Novel technique for thermal lens measurement in commonly used optical components.
The absorption of light in transmissive optics cause a thermally induced effect known as thermal lensing. This effect provokes an often undesired change of a laser beam transmitted by the optic. In this paper we present a measurement method that allows us to determine thermal lensing in commonly used optical components. The beam influenced by the thermal lens is expanded into the eigenmodes of an optical cavity, and its modal content is analyzed in the eigenbasis of the cavity. The measured quantity depends neither on beam parameters nor on the position of the optical component under investigation. This method allows, to our knowledge, for the first time the direct measurement of the mode conversion coefficient |ε(2)| of the thermal lens.


Severo Ochoa and the biomedical sciences in Spain under Franco, 1959-1975.
The influence of Severo Ochoa in the establishment of biochemistry and molecular biology in Spain is the central topic of this essay. From the time he was awarded the Nobel Prize in Physiology or Medicine in 1959, Ochoa's links with Spanish scientists and top authorities in education and science became instrumental to the development of these areas in the country of his birth. Ochoa's influence is analyzed through investigation of three "events": the reception of the award in Spain and some of its immediate consequences; his role in the VIth Meeting of the Federation of European Biochemical Societies, held in Madrid in 1969; and the international scientific symposium, held in Madrid and Barcelona, that celebrated his seventieth birthday in 1975. After an account of Ochoa's biography up to 1959, analysis of these events shows that Ochoa's influence cannot be understood without taking into account the political and scientific context of its reception.


Iodine nutrition status in Spain Needs for the future.
Although iodine nutrition in Spain has improved in recent years, the problem is not completely resolved. It is necessary that health institutions establish measures to ensure an adequate iodine nutrition of the population, especially among the highest risk groups (children and adolescents, women of childbearing age, pregnant women and nursing mothers). A low salt intake should be advised, but it should be iodized. It is also imperative that food control agencies establish effective control over adequate iodization of salt. Indicators on iodine nutrition should be included in future health surveys. The EUthyroid study and the Krakow Declaration on iodine nutrition provide an opportunity to set up a pan-European plan for the prevention of iodine deficiency that should be considered and used by health authorities.


Clinical and Pregnancy Outcomes of Coronavirus Disease 2019 Among Hospitalized Pregnant Women in the United States.
Pregnant women with coronavirus disease 2019 (COVID-19) may be at greater risk of poor maternal and pregnancy outcomes. This retrospective analysis reports clinical and pregnancy outcomes among hospitalized pregnant women with COVID-19 in the United States. The Premier Healthcare Database-Special Release was used to examine the impact of COVID-19 among pregnant women aged 15-44 years who were hospitalized and who delivered compared with pregnant women without COVID-19. Outcomes evaluated were COVID-19 clinical progression, including the use of supplemental oxygen therapy, intensive care unit admission, critical illness, receipt of invasive mechanical ventilation/extracorporeal membrane oxygenation, maternal death, and pregnancy outcomes, including preterm delivery and stillbirth. Overall, 473 902 hospitalized pregnant women were included, 8584 (1.8%) of whom had a COVID-19 diagnosis (mean age = 28.4 [standard deviation = 6.1] years; 40% Hispanic). The risk of poor clinical and pregnancy outcomes was greater among pregnant women with COVID-19 compared with pregnant women without a COVID-19 diagnosis in 2020; the risk of poor clinical and pregnancy outcomes increased with increasing age. Hispanic and Black non-Hispanic women were consistently observed to have the highest relative risk of experiencing poor clinical or pregnancy outcomes across all age groups. Overall, COVID-19 had a significant negative impact on maternal health and pregnancy outcomes. These data help inform clinical practice and counseling to pregnant women regarding the risks of COVID-19. Clinical studies evaluating the safety and efficacy of vaccines against severe acute respiratory syndrome coronavirus 2 in pregnant women are urgently needed.


Dynamic energy landscape view of coupled binding and protein conformational change: induced-fit versus population-shift mechanisms.
Allostery, the coupling between ligand binding and protein conformational change, is the heart of biological network and it has often been explained by two representative models, the induced-fit and the population-shift models. Here, we clarified for what systems one model fits better than the other by performing molecular simulations of coupled binding and conformational change. Based on the dynamic energy landscape view, we developed an implicit ligand-binding model combined with the double-basin Hamiltonian that describes conformational change. From model simulations performed for a broad range of parameters, we uncovered that each of the two models has its own range of applicability, stronger and longer-ranged interaction between ligand and protein favors the induced-fit model, and weaker and shorter-ranged interaction leads to the population-shift model. We further postulate that the protein binding to small ligand tends to proceed via the population-shift model, whereas the protein docking to macromolecules such as DNA tends to fit the induced-fit model.


Use of degenerate primers and touchdown PCR for construction of cDNA libraries.
Optimized construction of low-redundancy cDNA mini-libraries using low-stringency RT-PCR is described cDNAs are generated using arbitrary consensus-degenerate hybrid oligonucleotide primers and nanogram amounts of Schistosoma mansoni mRNA. A number of conditions such as temperature and salt concentration are combined to create balanced, low-stringency conditions that permit a normalized amplification of a diversified, random set of sequences. On average, 350 different sequences are obtained per mini-library, which represents a significantly higher diversity of messages per library when compared to previously published conditions (ie., 20-40 sequences/ mini-library). The optimized high-throughput approach described here is likely to help in the discovery of expressed genes in any complex organism.


The nucleoprotein of Marburg virus is target for multiple cellular kinases.
The nucleoprotein (NP) of Marburg virus is phosphorylated at serine and threonine residues in a ratio of 85:15, regardless of whether the protein is isolated from virions or from eukaryotic expression systems. Phosphotyrosine is absent. Although many potential phosphorylation sites are located in the N-terminal half of NP, this part of the protein is not phosphorylated. Analyses of phosphorylation state and phosphoamino acid content of truncated NPs expressed in HeLa cells using the vaccinia virus T7 expression system led to the identification of seven phosphorylated regions (region I*, amino acids 404-432; II*, amino acids 446-472; III*, amino acids 484-511; IV*, amino acids 534-543; V*, amino acid 549; VI*, amino acids 599-604; and VII*, amino acid 619) with a minimum of seven phosphorylated amino acid residues located in the C-terminal half of NP. All phosphothreonine residues and consensus recognition sequences for protein kinase CKII are located in regions I*-V*. Regions VI* and VII* contain only phosphoserine with three of four serine residues in consensus recognition motifs for proline-directed protein kinases. Mutagenesis of proline-adjacent serine residues to alanine or aspartic acid did not influence the function of NP in a reconstituted transcription/replication system; thus it is concluded that serine phosphorylation in the most C-terminal part of NP is not a regulatory factor in viral RNA synthesis.


Iron deficiency and marginal vitamin A deficiency affect growth, hematological indices and the regulation of iron metabolism genes in rats.
Iron deficiency and marginal vitamin A (VA) deficiency frequently coexist and affect billions of people, mostly children and women, worldwide. The effects of these micronutrient deficiencies alone and in combination on hematologic, biochemical and molecular indices of iron and VA status were investigated in a 2 x 2 randomized blocked study conducted in growing male Sprague-Dawley rats. From 3-8 wk of age, rats were fed one of four purified diets that were either adequate or restricted in iron (Fe) and adequate or marginal in VA: (+)Fe(+)VA, 20.3 and 0.367 micro g/g, respectively, denoted control diet; (-)Fe(+)VA, 3.34 and 0.405 micro g/g; (+)FeVA(m), 22.2 and 0.051 micro g/g; or (-)FeVA(m), 3.03 and 0.055 micro g/g. Weight-matched rats fed adequate micronutrients were included to control for possible confounding effects of Fe deficiency on growth and feed efficiency. Iron restriction reduced (P < 0.05) weight gain, feed efficiency, blood hemoglobin and hematocrit. Plasma and liver iron and plasma transferrin saturation were reduced by approximately 50%, whereas liver transferrin mRNA and protein and transferrin receptor mRNA were elevated, as was liver ferritin light-chain protein and light-chain mRNA. Liver heavy-chain ferritin mRNA, hemopexin, ceruloplasmin and cellular retinol-binding protein mRNA were not affected by iron or VA restriction. Although marginal VA deficiency did not exacerbate indices of poor iron status during iron deficiency, iron deficiency was associated with lower plasma retinol and elevated liver VA concentrations. These results are consistent with an impaired mobilization of liver retinol during iron deficiency as well as multiple alterations in iron metabolism.


Effect of alkyl substituents on photorelease from butyrophenone derivatives.
Photolysis of 1a yields 4a in argon-saturated methanol, whereas 1b is photostable. Laser flash photolysis of 1a in acetonitrile shows formation of biradical 2a (lambda(max) = 340 nm, tau = approximately 60 ns), which undergoes intersystem crossing to form Z-3a (lambda(max) = 380 nm, tau = 270 ns) and E-3a (lambda(max) = 380 nm, tau = 300 ms). Z-3a regenerates the starting material, whereas E-3b undergoes intramolecular lactonization to release the alcohol moiety and form 4a. Similar laser flash photolysis of 1b shows formation of biradical 2b (lambda(max) = 340 nm, tau = 1.9 micros in acetonitrile), which is longer-lived than 2a is. However, 2b only undergoes intersystem crossing to form Z-3b (lambda(max) = 380 nm, tau = 4.3 micros). Calculations demonstrate that intramolecular pseudo hydrogen bonding between the OH moiety and the radical centered on the isopropyl carbon in 2b and the bulkiness of the isopropyl group prevent the necessary rotation to form E-3b. In comparison, 2a does not form an intramolecular pseudo hydrogen bond between the methylene radical center and the OH group, and as a consequence, it undergoes intersystem crossing to form both E- and Z-3a.


Dexamethasone and lactoferrin induced PMN-MDSCs relieved inflammatory adverse events of anti-cancer therapy without tumor promotion.
In this era of immune checkpoint inhibitors, inflammatory adverse events of anti-cancer therapies continue to pose a major challenge. Glucocorticoids, as the mainstay, were limited by serious side effects. Glucocorticoids induce myeloid-derived suppressor cells (MDSCs), and lactoferrin-induced polymorphonuclear MDSCs (PMN-MDSCs) were shown to relieve inflammatory conditions. Combined treatment with dexamethasone (DXM) and lactoferrin increased the generation of PMN-MDSCs in vitro (DXM/lactoferrin PMN-MDSCs) compared to DXM or lactoferrin treatment alone. DXM/lactoferrin PMN-MDSCs were distinct from tumor PMN-MDSCs in vivo with regard to gene expression profiles. DXM upregulated the myeloid cell response to lactoferrin by inducing the lactoferrin receptor Lrp1. DXM/lactoferrin PMN-MDSCs presented anti-bacterial capability, increased PGE2 production, increased survival capability, and decreased tumor tissue homing. Transfer of DXM/lactoferrin PMN-MDSCs relieved cisplatin-induced acute kidney failure, bleomycin-induced interstitial pneumonia, and allergic pneumonitis effectively without promoting tumor development. Our study shows that DXM/lactoferrin PMN-MDSCs are a promising cell therapy for inflammatory adverse events of anti-cancer therapies.


Structural and functional characterization of HP0377, a thioredoxin-fold protein from Helicobacter pylori.
Maturation of cytochrome c is carried out in the bacterial periplasm, where specialized thiol-disulfide oxidoreductases provide the correct reduction of oxidized apocytochrome c before covalent haem attachment. HP0377 from Helicobacter pylori is a thioredoxin-fold protein that has been implicated as a component of system II for cytochrome c assembly and shows limited sequence similarity to Escherichia coli DsbC, a disulfide-bond isomerase. To better understand the role of HP0377, its crystal structures have been determined in both reduced and partially oxidized states, which are highly similar to each other. Sedimentation-equilibrium experiments indicate that HP0377 is monomeric in solution. HP0377 adopts a thioredoxin fold but shows distinctive variations as in other thioredoxin-like bacterial periplasmic proteins. The active site of HP0377 closely resembles that of E. coli DsbC. A reductase assay suggests that HP0377 may play a role as a reductase in the biogenesis of holocytochrome c553 (HP1227). Binding experiments indicate that it can form a covalent complex with HP0518, a putative L,D-transpeptidase with a catalytic cysteine residue, via a disulfide bond. Furthermore, physicochemical properties of HP0377 and its R86A variant have been determined. These results suggest that HP0377 may perform multiple functions as a reductase in H. pylori.


Molecular classification of endometrial carcinoma, is it the new era of precision medicine?
For many years endometrial cancer has been subdivided into oestrogen - dependent (type I) and oestrogen - independent (type II), according to classical Bokhman classification. Histopathological evaluation including type and grade of tumour, along with clinical factors have been considered as very important prognostic factors that impact treatment decision. However, histologically similar tumours may have different outcomes. Recent molecular findings and new histopathological parameters have given new concept on risk stratification. The Cancer Genome Atlas Research Network (TCGA) of tumours have brought new insights into endometrial cancer management. Four molecular subgroups have been described: POLE ultramutated (POLE mut), p53 mutant (p53abn), mismatch repair deficient (MMRd) and non-specific molecular profile (NSMP). This new subdivision has been recently introduced in the European risk stratification system.


Synthesis, characterization, and in vitro evaluation of new Ibuprofen polymeric prodrugs based on 2-hydroxypropyl methacrylate.
The present research work describes the synthesis and evaluation of new acrylic-type polymeric systems having degradable ester bonds linked to ibuprofen as materials for drug delivery. Ibuprofen was linked to 2-hydroxy-propyl methacrylate by an activated ester methodology in a one-pot procedure with a high yield. The resulting material was copolymerized with either 2-hydroxyethyl methacrylate or methyl methacrylate (in 1:3 mole ratios) by the free radical polymerization method, utilizing azoisobutyronitrile at 65-70 °C. The characterization of the resulting products by FTIR, (1)H NMR, (13)C NMR, DSC, and elemental analysis confirmed their synthesis successfully. Ibuprofen release from the obtained polymers was preliminarily evaluated at different buffered solutions (pH 1, 7.4, and 10) into dialysis bags to show the capacity of prodrugs to release the drug under hydrolytic conditions. Detection of hydrolysis by UV spectroscopy at selected intervals showed that the drug can be released by selective hydrolysis of the ester bond at the side of the drug moiety. The release profiles indicated that the hydrolytic behavior of polymers is strongly based on the polymer hydrophilicity and the pH value of the hydrolysis solution. The results suggest that these polymers could be useful in controlled release systems.


Intersegmental transfer of sensory signals in the stick insect leg muscle control system.
Intersegmental coordination during locomotion in legged animals arises from mechanical couplings and the exchange of neuronal information between legs. Here, the information flow from a single leg sense organ of the stick insect Cuniculina impigra onto motoneurons and interneurons of other legs was investigated. The femoral chordotonal organ (fCO) of the right middle leg, which measures posture and movement of the femur-tibia joint, was stimulated, and the responses of the tibial motoneuron pools of the other legs were recorded. In resting animals, fCO signals did not affect motoneuronal activity in neighboring legs. When the locomotor system was activated and antagonistic motoneurons were bursting in alternation, fCO stimuli facilitated transitions from flexor to extensor activity and vice versa in the contralateral leg. Following pharmacological treatment with picrotoxin, a blocker of GABA-ergic inhibition, the tibial motoneurons of all legs showed specific responses to signals from the middle leg fCO. For the contralateral middle leg we show that fCO signals encoding velocity and position of the tibia were processed by those identified local premotor nonspiking interneurons known to contribute to posture and movement control during standing and voluntary leg movements. Interneurons received both excitatory and inhibitory inputs, so that the response of some interneurons supported the motoneuronal output, while others opposed it. Our results demonstrate that sensory information from the fCO specifically affects the motoneuronal activity of other legs and that the layer of premotor nonspiking interneurons is a site of interaction between local proprioceptive sensory signals and proprioceptive signals from other legs.


Deposition of collagen VI in the extracellular matrix during mouse embryogenesis correlates with expression of the alpha 3(VI) subunit gene.
Collagen VI is a microfibrillar component of the extracellular matrix that is predicted to have an important structural role in matrix organization and a biological function in mediating cell-matrix interactions. Secreted collagen VI molecules are composed of three distinct subunits, the alpha 1(VI), alpha 2(VI), and alpha 3(VI) chains. To determine when, and in which tissues, collagen VI is likely to have a role in embryonic processes, we have analyzed the expression patterns of the three subunit chains during postimplantation mouse development by reverse transcriptase-PCR (RT-PCR), in situ hybridization, and immunofluorescence. No collagen VI protein could be detected in the mouse embryo until Day 11.5 of gestation, when low levels were localized within the mesoderm layer of the visceral yolk sac, the subepidermal matrix of branchial arches, and the vessel wall of the dorsal aorta. Levels of collagen VI mRNA and protein increased during the period from Days 12.5 to 14.5 in the visceral yolk sac, subepidermal mesenchyme, lung, gut, meninges, muscle, perichondrium, and vertebral column. The cartilage matrix of ribs and developing long bones was not stained with collagen VI antisera, but pericellular staining of chondrocytes was seen in both tissues. Low levels of collagen VI mRNA and protein were seen in the fetal liver except for the connective tissue of the liver capsule, which was highly stained. Collagen VI was first detected at significant levels in the developing heart on Day 14.5. These data demonstrate a tissue-specific onset of collagen VI synthesis and deposition in the extracellular matrix of developing mouse embryos at a much later stage of development than that reported for fibronectin or collagen I. Sensitive RT-PCR assays showed that alpha 1(VI) and alpha 2(VI) mRNAs were amplified from extracts of embryonic tissues as early as Day 7.5, while alpha 3(VI) mRNA was not detected until Day 10.5. Expression of the alpha 3(VI) gene immediately preceded the appearance of collagen VI protein in embryonic tissues. This correlation is consistent with the proposal that expression of alpha 3(VI) chains regulates the formation and secretion of collagen VI trimers and collagen VI matrix deposition during development.


Protein/polysaccharide-based scaffolds mimicking native extracellular matrix for cardiac tissue engineering applications.
Tissue engineering has emerged as a viable approach to treat disease or repair damage in tissues and organs. One of the key elements for the success of tissue engineering is the use of a scaffold serving as artificial extracellular matrix (ECM). The ECM hosts the cells and improves their survival, proliferation, and differentiation, enabling the formation of new tissue. Here, we propose the development of a class of protein/polysaccharide-based porous scaffolds for use as ECM substitutes in cardiac tissue engineering. Scaffolds based on blends of a protein component, collagen or gelatin, with a polysaccharide component, alginate, were produced by freeze-drying and subsequent ionic and chemical crosslinking. Their morphological, physicochemical, and mechanical properties were determined and compared with those of natural porcine myocardium. We demonstrated that our scaffolds possessed highly porous and interconnected structures, and the chemical homogeneity of the natural ECM was well reproduced in both types of scaffolds. Furthermore, the alginate/gelatin (AG) scaffolds better mimicked the native tissue in terms of interactions between components and protein secondary structure, and in terms of swelling behavior. The AG scaffolds also showed superior mechanical properties for the desired application and supported better adhesion, growth, and differentiation of myoblasts under static conditions. The AG scaffolds were subsequently used for culturing neonatal rat cardiomyocytes, where high viability of the resulting cardiac constructs was observed under dynamic flow culture in a microfluidic bioreactor. We therefore propose our protein/polysaccharide scaffolds as a viable ECM substitute for applications in cardiac tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 769-781, 2018.


[A study on the role of the transmembrane V to VI regions of the human luteinizing hormone receptor in its constitutive activation].
Gonadotropin (luteinizing hormone: LH, and follicle stimulating hormone: FSH) receptors are members of the seven-transmembrane (TM) receptor family. Recently, point mutations have been identified in the LH receptor gene, leading to constitutive activation (ligand-independent activation) of the receptor in patients with familial male-limited precocious puberty. Most of such point mutations were found in the coding region consisting of TM V, VI and the intracellular loop 3 (i3). Because the primary structures of gonadotropin receptors are highly conserved, in vitro mutagenesis was performed to introduce the amino acid substitution in the i3 domain (Asp-->Gly) of FSH receptor, corresponding to the point mutation leading to constitutive activation in LH receptor. However, the FSH receptor mutant showed almost similar basal cAMP production to wild type counterpart. For investigation of this difference in constitutive activation between two receptors, the author designed chimeric LH/FSH receptors with or without an amino acid substitution in the i3 (Asp-->Gly) to identify the region in the LH receptor which is important for its constitutive activation. Introduction of the amino acid substitution into a chimeric receptor containing only TM V to VI from the LH receptor led to major increases in ligand-independent cAMP production. Furthermore, the chimeric receptor with only TM V and VI derived from the LH receptor was constitutively activated by the amino acid substitution in the i3 from the FSH receptor. These results that interactions between TM V and VI of the FSH receptor are essential for maintaining the receptor conformation constrained, whereas interactions between these domains of the LH receptor are required for constitutive activation by the amino acid substitution in the i3.


[Expression of nerve growth factor and its receptor during osteoinduction of recombined human bone morphogenetic protein 2].
To identify the expression of nerve growth factor (NGF) and its high affinity receptor (tyrosine kinase receptor A, TrkA) during bone induction by recombined human bone morphogenetic protein 2 (rhBMP-2) by immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) and to discuss the role of NGF on the bone induction of the BMP. Thirty-six ICR mice were divided into the experimental group and the control group at random. rhBMP-2 /collagen sponge and collagen sponge were implanted into the right thigh muscle pouches of the mice in the experimental group and the control group, respectively. The tissues in the implanted site of the two groups were removed on the 7th, 14th and 21st day after the implantation. Histological, immunohistochemical and RT-PCR analyses were performed to detect osteoinductive effects of rhBMP-2 and the expression of NGF and TrkA. Gross observation showed that a solid lump was found in the right thigh in the experimental group on the 7th day and became harder on the 14th and 21st day, which was not found in the control group. rhBMP-2 /collagen sponge displayed a potent ability to induce bone formation, while immunostaining for NGF and TrkA was observed in the course of osteoinduction by rhBMP-2. On the 7th day in the experimental group, NGF positive immunostaining reached the peak in the stage of chondrogenesis and there were a large number of cells expressing NGF, including fibroblasts, chondroblasts, chondrocytes, and osteoblasts; then, there was a decrease in the number of the positive cells and in the intensity of immunostaining on the 14th and 21st day. Staining of TrkA was similar to that of NGF. The expression level of the mRNA of NGF during the course of bone induction peaked 7 days rhBMP-2/collagen is a kind of satisfactory osteoinductive after the implantation and then decreased. material, and many different kinds of cells induced by rhBMP-2 can express NGF and TrkA, which suggests that NGF may play an important role in the osteogenesis initiated by exogenous BMP through direct and indirect pathways.


Dynamics of Donor-Derived Cell-Free DNA at the Early Phase After Pediatric Kidney Transplantation: A Prospective Cohort Study.
Background: Donor-derived cell-free DNA (ddcfDNA) has been suggested as an indicator of allograft injury in adult and pediatric kidney transplantation (KTx). However, the dynamics of ddcfDNA in pediatric KTx have not been investigated. In addition, it has not been demonstrated whether donor-recipient (D/R) size mismatch affect ddcfDNA level. Methods: Pediatric KTx recipients with a single donor kidney were enrolled and followed up for 1 year. ddcfDNA, calculated as a fraction (%) in the recipient plasma, was examined longitudinally within 3 months post-transplant. D/R size mismatch degree was described as D/R height ratio. The 33rd percentile of D/R height ratio (0.70) was used as the cut-off to divide the patients into low donor-recipient height ratio group (<0.70) and high donor-recipient height ratio group (≥0.70). The dynamics of ddcfDNA were analyzed and the impact factors were explored. Stable ddcfDNA was defined as the first lowest ddcfDNA. ddcfDNA flare-up was defined as a remarkable elevation by a proportion of >30% from stable value with a peak value >1% during elevation. Results: Twenty-one clinically stable recipients were enrolled. The median D/R height ratio was 0.83 (0.62-0.88). It took a median of 8 days for ddcfDNA to drop from day 1 and reach a stable value of 0.67% (0.46-0.73%). Nevertheless, 61.5% patients presented ddcfDNA>1% at day 30. Besides, 81.0% (17/21) of patients experienced elevated ddcfDNA and 47.6% (10/21) met the standard of ddcfDNA flare-up. Donor-recipient height ratio was an independent risk factor for ddcfDNA flare-up (odds ratio = 0.469 per 0.1, 95% CI 0.237-0.925, p = 0.029) and low donor-recipient height ratio (<0.70) was found to increase the risk of flare-up occurrence (odds ratio = 15.00, 95% CI 1.342-167.638, p = 0.028). Conclusions: ddcfDNA rebounds in many stable pediatric KTx recipients without rejection. This may be induced by significant D/R size mismatch and may affect its diagnostic performance at the early phase after pediatric KTx in children.


Matrix metalloproteinase, vitamin A and methylprednisolone effects on experimentally induced amyloid arthropathy.
We evaluated the role of some matrix metalloproteinases (MMPs) in enhancing the effect of vitamin A and the inhibiting effect of methylprednisolone on amyloid arthropathy in brown layer chicks. We used 100 one-day-old Isa brown layer chicks. The chicks were allocated to one of four groups as follows: negative control group (I), vitamin A group (II), positive control group (III) and methylprednisolone group (IV). Amyloid arthropathy was induced by injections of complete Freund's adjuvant into the left intertarsal joints of the chicks. Serum vitamin A and tissue MMP (MMP-1, MMP-2, MMP-9) levels were measured and differences among the groups were investigated. Serum vitamin A rates (μg/dl) were: 63.57 ± 4.10, 47.13 ± 10.62, 53.26 ± 10.79, 98.48 ± 8.20 in groups I, II, III and IV, respectively (p < 0.001). MMP-1, MMP-2 and MMP-9 levels were evaluated in tissues from the chickens with amyloid arthropathy. Methylprednisolone significantly suppressed the release of MMP-1 and MMP-2, and increased the release of MMP-9 in birds with amyloid arthropathy. In addition, vitamin A significantly increased the release of MMP-1, MMP-2 and MMP-9.


Arthroscopic findings in the overhand throwing athlete: evidence for posterior internal impingement of the rotator cuff.
The purpose of this article is to describe the outcome of an arthroscopic examination and the pathology in symptomatic shoulders of 41 professional overhand throwing athletes. With the arm in the position of the relocation test, 100% of the subjects had either contact between the rotator cuff undersurface and the posterosuperior glenoid rim or osteochondral lesions. Other key findings included undersurface cuff fraying in 93%, posterosuperior labral fraying in 88%, and anterior labral fraying in 36% of the subjects. This study supports the concept of impingement of the posterior cuff undersurface with the posterosuperior glenoid rim in the overhand throwing athlete with shoulder pain.


Endoscopic endonasal transsphenoidal approach for pituitary adenomas: technical aspects and report of casuistic.
Analyse technical aspects, effectiveness and morbidity of the endoscopic endonasal transphenoidal approach for pituitary adenomas. From January 2005 to September 2008, 30 consecutive patients underwent endoscopic endonasal resection of pituitary adenomas with a follow up from 3 to 36 months. Their medical charts were retrospectively analysed. There were 18 women and 12 men, mean age 44 years (range 17-65 yr). Among the 30 patients, 23 had macroadenomas and 7 microadenomas. Twelve patients had non-functioning tumors, 9 had ACTH-secreting tumors, 8 had GH-secreting tumors and 1 prolactinoma. Complete resection and hormonal control was achieved in all microadenomas. Macroadenomas were completely removed in 6 patients, subtotal resection in 6 and partial resection in 11. Three patients had diabetes insipidus and 5 had CSF leaks treated with lumbar drainage. The endonasal endoscopic approach for pituitary tumors is effective and has low morbidity.


Effect of High-Flow Nasal Cannula on Expiratory Pressure and Ventilation in Infant, Pediatric, and Adult Models.
Heated and humidified high-flow nasal cannula (HFNC) is a widely used form of respiratory support; however, data regarding optimal flows for a given patient size or disease state are lacking. A comprehensive study of the physiologic effects of HFNC is needed to better understand the mechanisms of action. The objective of the current study was to quantify the effect of HFNC settings in age-specific, anatomically correct nasal airways and spontaneously breathing lung models. We hypothesized that there is an effect of flow on pressure and ventilation. Three-dimensionally printed upper airway models of a preterm neonate, term neonate, toddler, small child, and adult were affixed to the ASL 5000 test lung to simulate spontaneous breathing with age-appropriate normal ventilation parameters. CO2 was introduced to simulate profound hypercapneic respiratory failure with an end-tidal partial pressure of carbon dioxide (PETCO2 ) of 90 ± 1 mm Hg. Two commercially available HFNC systems were applied to the airway models, and PEEP, inspired CO2, and exhaled CO2 (PETCO2 ) were recorded for 6 min across a range of flow. Increasing HFNC flow provided a non-linear increase in PEEP in closed-mouth models, with maximum tested flows generating 6 cm H2O in the preterm neonate to 20 cm H2O in the small child. Importantly, PEEP decreased by approximately 50% in open-mouth models. Increasing HFNC flow improved expiratory CO2 elimination to a certain point, above which continued increases in flow had minimal additional effect. This change point ranged from 4 L/min in the preterm neonate to 10 L/min in the small child. These findings may help clinicians understand the effects of HFNC at different settings and may inform management guidelines for patients with respiratory failure.


Cholesterol screening in children: should obesity be a risk factor?
To determine whether obesity should be added to the current American Academy of Pediatrics (AAP) criteria for cholesterol screening in childhood, the charts of 99 children referred for evaluation of either hypercholesterolemia (n = 53) or obesity (n = 45) were reviewed. Compared with obese children, nonobese hypercholesterolemic subjects were younger (8.4 vs 11.4 years) and had lower mean body mass index and % ideal body weight. Frequency of elevated (> 90th percentile for age) total and low-density lipoprotein cholesterols were similar in both groups. Fifty-three of 65 children who met the current AAP criteria were hypercholesterolemic, however, 23/76 hypercholesterolemic children failed to satisfy these screening criteria. Thirty-six of 45 obese children had cholesterol levels > 90th percentile, suggesting increased risk for hypercholesterolemia in this group. If obesity was added to the AAP criteria, 66/80 hypercholesterolemic subjects would have been identified. These modified criteria, vs AAP standards, significantly improved both their sensitivity (70 vs 87%, p < 0.02) and negative predictive value (45 vs 30%, p < 0.02). Pending further studies in larger pediatric populations, these data indicate that obesity should be considered a risk factor for hypercholesterolemia in childhood, and we recommend modifying the AAP screening criteria to include obese children.


Using technology to engage and educate youth.
Given that information age technologies are as natural to the Net generation as breathing, it is time to provide learning experiences that maximize their use in schools. The authors argue that integrating technology into learning is central to creating the meaningful learning opportunities needed to engage and motivate youth today. To achieve this potential, however, technology must support a new paradigm of teaching and learning. This chapter discusses common elements and challenges to implementing pioneering high school reform models that promote one-to-one student-computer ratios, collaborative learning environments, problem- and project-based instruction, and interdisciplinary teaching.


Predictive value of thrombocytopaenia in the diagnosis of dengue infection in outpatient settings.
Thrombocytopaenia is often relied upon as an important criterion for the diagnosis of dengue infection among patients presenting with an acute non-specific febrile illness. This study was aimed to assess usefulness of thrombocytopaenia in the diagnosis of acute dengue virus infection. This was a clinic based prospective cohort study from May to November 2003. Consecutive patients presenting with acute non-specific febrile illness of less than two weeks were selected from two urban primary care centres. We did full blood count examination (FBC) on the day of visit and dengue serology on day five of illness for all patients enrolled. We repeated the FBC examination for patients who had initial normal platelet counts. Thrombocytopaenia was defined as platelet count < 150 X 10(9)/L. Eighty-seven patients enrolled in the study. Complete data was available for 73 patients. The prevalence of acute dengue virus infection was 27.6%. The sensitivity and specificity were 88% and 71% respectively. The likelihood of acute dengue infection in the presence of thrombocytopaenia was 2.52 and likelihood of not having dengue infection in normal platelet count patients was 5.22. Thrombocytopaenia has fair predictive value in diagnosing acute dengue virus infection. It was more useful to exclude than to diagnose dengue infection.


Caveolin-1 Promotes the Imbalance of Th17/Treg in Chronic Obstructive Pulmonary Disease by Regulating Hsp70 Expression.
To investigate whether the expression of Hsp70 is associated with Cav-1 in promoting the imbalance of Th17/Treg cells in COPD. The plasma Cav-1, Hsp70 expression were quantified by enzyme-linked immunosorbent assay (ELISA). The frequencies of circulating Th17, Treg cells and Th17/Treg ratio were determined by flow cytometry. Peripheral blood mononuclear cells (PBMCs) from subjects were transfected with Cav-1 or control plasmids and Hsp70 plasmid. We found that Cav-1 expression was lower but the levels of Hsp70 and Th17 cells were higher in COPD than in healthy control (HC). Hsp70 expressions were positively correlated with Cav-1 levels, Th17 cells, and Th17/Treg ratio in COPD but not in HC. Cav-1 over-expression resulted in an increase in Hsp70 and Th17 levels. Suppressing Hsp70 expressing by small interfering RNA (siRNA), the decline of Th17 frequency was observed in Cav-1-overexpressed PBMCs. Collectively, our results illuminate that Cav-1 contributes to the imbalance of Th17/Treg through potentially regulating Hsp70 expression.


Shuyu pills inhibit immune escape and enhance chemosensitization in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is characterized by dysregulation of the immune microenvironment and the development of chemoresistance. Specifically, expression of the programmed cell death protein 1 (PD-1)/programmed cell death 1 ligand 1 (PD-L1) axis, an immune checkpoint, may lead to tumour immune escape, resulting in disease progression. The latest research shows that tumour immune escape may be caused by the upregulation of PD-L1 mediated by hypoxia-inducible factor-1 alpha (HIF-1α), and simultaneous inhibition of HIF-1α and PD-L1 has the potential to enhance the host's antitumour immunity. Moreover, inhibition of the PD-1/PD-L1 axis may mitigate tumour chemoresistance. Shuyu pills (SYPs) contain immunity-enhancing and antitumour components, making them a potential HCC treatment. To investigate the efficacy of SYPs for HCC treatment via simultaneous HIF-1α and PD-L1 inhibition and the mechanism involved. A subcutaneous xenograft tumour model was first established in BALB/c nude mice by the subcutaneous injection of 1 × 107 SMMC-7721 cells. Male mice (male, 5 weeks old; n = 24) were then randomly divided into the following four groups (n = 6): Control (0.9% normal saline), SYP (200 mg/kg), SYP + cisplatin (DDP) (200 mg/kg + 5 mg/kg DDP weekly via intraperitoneal injection), and DDP (5 mg/kg cisplatin weekly via intraperitoneal injection). The dose of saline or SYPs for the indicated mouse groups was 0.2 mL/d via intragastric administration. The tumour volumes and body weights of the mice were measured every 2 d. The mice were euthanized by cervical dislocation after 14 d of continuous treatment, and the xenograft tissues were excised and weighed. Western blot assays were used to measure the protein expression of HIF-1α, PD-1, PD-L1, CD4+ T cells, and CD8+ T cells in HCC tumours from mice. Quantitative reverse transcription polymerase chain reaction was used for real-time quantitative detection of PD-1, PD-L1, and HIF-1α mRNA expression. An immunofluorescence assay was conducted to examine the expression of CD4+ T cells and CD8+ T cells. Compared to mice in the control group, those in the SYP and SYP + DDP groups exhibited reduced tumour volumes and tumour weights. Moreover, the protein and mRNA expression levels of the oncogene HIF-1α and that of the negative immunomodulatory factors PD-1 and PD-L1 were decreased in both the SYP and SYP + DDP groups, with the decrease effects being more prominent in the SYP + DDP group than in the SYP group (HIF-1α protein: Control vs SYP, P = 0.0129; control vs SYP + DDP, P = 0.0004; control vs DDP, P = 0.0152, SYP + DDP vs DDP, P = 0.0448; HIF-1α mRNA: control vs SYP, P = 0.0009; control vs SYP + DDP, P < 0.0001; control vs DDP, P = 0.0003, SYP vs SYP + DDP, P = 0.0192. PD-1 protein: Control vs SYP, P = 0.0099; control vs SYP + DDP, P < 0.0001, SPY vs SYP + DDP, P = 0.0009; SYP + DDP vs DDP, P < 0.0001; PD-1 mRNA: control vs SYP, P = 0.0002; control vs SYP + DDP, P < 0.0001; control vs DDP, P = 0.0003, SPY vs SYP + DDP, P = 0.0003; SYP + DDP vs DDP, P = 0.0002. PD-L1 protein: control vs SYP, P < 0.0001; control vs SYP + DDP, P < 0.0001; control vs DDP, P < 0.0001, SPY vs SYP + DDP, P = 0.0040; SYP + DDP vs DDP, P = 0.0010; PD-L1 mRNA: Control vs SYP, P < 0.0001; control vs SYP + DDP, P < 0.0001; control vs DDP, P < 0.0001, SPY vs SYP + DDP, P < 0.0001; SYP + DDP vs DDP, P = 0.0014). Additionally, the quantitative and protein expression levels of CD4+ T cells and CD8+ T cells were simultaneously upregulated in the SYP + DDP group, whereas only the expression of CD4+ T cells was upregulated in the SYP group. (CD4+ T cell quantitative: Control vs SYP + DDP, P < 0.0001, SYP vs SYP + DDP, P = 0.0005; SYP + DDP vs DDP, P = 0.0002. CD4+ T cell protein: Control vs SYP, P = 0.0033; Control vs SYP + DDP, P < 0.0001; Control vs DDP, P = 0.0021, SYP vs SYP + DDP, P = 0.0004; SYP + DDP vs DDP, P = 0.0006. Quantitative CD8+ T cells: Control vs SYP + DDP, P = 0.0013; SYP vs SYP + DDP, P = 0.0347; SYP + DDP vs DDP, P = 0.0043. CD8+ T cell protein: Control vs SYP + DDP, P < 0.0001; SYP vs SYP + DDP, P < 0.0001; SYP + DDP vs DDP, P < 0.0001). Finally, expression of HIF-1α was positively correlated with that of PD-1/PD-L1 and negatively correlated with the expression of CD4+ T cells and CD8+ T cells. SYPs inhibit immune escape and enhance chemosensitization in HCC via simultaneous inhibition of HIF-1α and PD-L1, thus inhibiting the growth of subcutaneous xenograft HCC tumours.


Dose titration of intramuscular interferon beta-1a reduces the severity and incidence of flu-like symptoms during treatment initiation.
Flu-like symptoms (FLS) are common side effects of interferon beta (IFNβ) therapy and can negatively affect the willingness of patients with multiple sclerosis to initiate therapy. Although dose titration is commonly used to reduce the severity and incidence of IFNβ-related FLS during treatment initiation, these benefits have not been confirmed in a well controlled study. The objective of this randomized, dose-blinded, parallel-group study was to assess the effect of dose titration on the severity and incidence of FLS during the initial 8 weeks of once-weekly intramuscular (IM) IFNβ-1a administration. Healthy volunteers were randomized 1:1:1 to one of three IM IFNβ-1a regimens: 3-week titration (weekly quarter-dose increments over 3 weeks to full dose [30 µg]); 6-week titration (biweekly quarter-dose increments over 6 weeks to full dose); or no titration (full dose over 8 weeks). At weekly clinic visits, the severity of each FLS was rated 1 hour pre-injection and 4-6 hours and 12-15 hours post-injection. Study endpoints included post-injection change in FLS severity and post-injection FLS incidence (percentage of subjects with a ≥2-point increase in total FLS severity score) at each time point. Clinicaltrials.gov identifier: NCT01119677. Of 234 subjects enrolled, 194 (83%) completed the study. At 8 weeks, FLS severity was significantly reduced at both post-injection time points with 3-week titration (76% reduction at 4-6 hours, p < 0.001; 37% reduction at 12-15 hours; p < 0.001) and 6-week titration (50% reduction at 4-6 hours, p < 0.001; 32% reduction at 12-15 hours; p = 0.002) compared with no titration. The incidence of FLS was also significantly reduced at both time points with both titration regimens. Safety profiles for both titration regimens were consistent with the current IM IFNβ-1a label. Study limitations included that there is currently no validated assessment tool for evaluating the severity of FLS, that the study enrolled healthy volunteers, that different proportions of females were randomized to the 3-week-titration group than to the 6-week and no-titration groups, and that evaluation of the potential impact of titration on symptoms occurring substantially later after injection was not part of the study protocol. Dose titration during initiation of IM IFNβ-1a reduces FLS severity and incidence in healthy volunteers compared with no titration.


Toward a Defined Role for Occupational Therapy in Foster Care Transition Programming.
Youth who age out of the foster care system and transition to adulthood face challenges that are exacerbated by a history of trauma, severed relationships, and instability of living and educational placements. A review of the literature demonstrates poor outcomes overall for this population. Occupational therapists are positioned to meet the needs that arise during this time; however, a review of emerging roles for occupational therapists is necessary to describe how occupational therapists can best fulfill gaps in current programming. Through a review of the literature and a preliminary mixed-methods study, this paper establishes a direction for the inclusion of occupational therapy for youth aging out of foster care using the Person Environment Occupation Performance (PEOP) model as a structure. Federal, state, and local organizations provide resources to assist transitioning foster youth. However, there is a lack of collaborative, individualized, and evidence-based approaches reporting good outcomes. Specific occupational therapy interventions are suggested to delineate our role with this high-risk population during transition to independent living: both novel interventions and additions to current evidence-based programming.


A mathematical framework for describing and analysing gene regulatory networks.
This paper presents a mathematical framework for describing and analysing gene regulatory networks by autonomous differential equations. It represents an improvement on existing frameworks in that it may handle a wider range of gene regulatory mechanisms. Gene regulatory networks are frequently threshold-dominated, i.e. genes are activated only when the concentration of certain gene products lie between definite thresholds. Here, the concept of regulatory domain is introduced to describe these regions in the phase space. To each regulatory domain is associated an indicator function whose value is 1 inside and 0 outside the domain. The indicator functions thus reflect the logical structure of the network. The sharp borders between the regulatory domains may be smoothed by replacing the logical step functions by continuous sigmoids or so-called logoid functions. A logoid function coincides with the step function outside a narrow interval around the threshold, and rises continuously from 0 to 1 inside it. Using logoids, the task of finding steady states is considerably simplified. A list of regions in phase space comprising all steady states lying close to a threshold is obtained by examining a certain type of matrix called the Logoid-Jacobian. In addition, this matrix leads to the conditions necessary for stability of the steady states. External signals may be conveniently incorporated in the form of Boolean variables. Thus the framework is well suited for studying gene regulatory networks both in single cells and multicellular systems.


Mapping epistemic cultures and learning potential of participants in citizen science projects.
The ever-widening scope and range of global change and interconnected systemic risks arising from people-environment relationships (social-ecological risks) appears to be increasing concern among, and involvement of, citizens in an increasingly diversified number of citizen science projects responding to these risks. We examined the relationship between epistemic cultures in citizen science projects and learning potential related to matters of concern. We then developed a typology of purposes and a citizen science epistemic-cultures heuristic and mapped 56 projects in southern Africa using this framework. The purpose typology represents the range of knowledge-production purposes, ranging from laboratory science to social learning, whereas the epistemic-cultures typology is a relational representation of scientist and citizen participation and their approach to knowledge production. Results showed an iterative relationship between matters of fact and matters of concern across the projects; the nexus of citizens' engagement in knowledge-production activities varied. The knowledge-production purposes informed and shaped the epistemic cultures of all the sampled citizen science projects, which in turn influenced the potential for learning within each project. Through a historical review of 3 phases in a long-term river health-monitoring project, we found that it is possible to evolve the learning curve of citizen science projects. This evolution involved the development of scientific water monitoring tools, the parallel development of pedagogic practices supporting monitoring activities, and situated engagement around matters of concern within social activism leading to learning-led change. We conclude that such evolutionary processes serve to increase potential for learning and are necessary if citizen science is to contribute to wider restructuring of the epistemic culture of science under conditions of expanding social-ecological risk.


Field-dependent Brownian relaxation dynamics of a superparamagnetic clustered-particle suspension.
The distinguishable Brownian relaxation dynamics of a clustered-particle system of superparamagnetic iron oxide nanoparticle suspension compared to that of a dispersed-particle system has been experimentally investigated through characterization of the frequency and field strength dependences of complex magnetic susceptibility. We confirmed that the application of low sinusoidal magnetic field strength enables cluster rotation instead of individual particle rotations. Furthermore, we found that the cluster rotation was altered to individual particle rotations in higher field strength, resulting in a shorter Brownian relaxation time, which suggests a change in the hydrodynamic volume. This evolutional relaxation behavior was associated with a change in the fitting parameter which satisfies the empirical model of relaxation and further represents the significance of interparticle interactions in defining the nonlinearity of the magnetization response.


Expression of vascular endothelial growth factor A isoforms is dysregulated in women with endometriosis.
Angiogenesis is a critical step in the development of ectopic lesions during endometriosis. Although total vascular endothelial growth factor (VEGF) A is elevated in the peritoneal fluid of women with endometriosis, there are contradictory reports on how levels of total endometrial VEGFA are altered in this disease. Furthermore, limited research is available on different VEGFA isoforms in women with endometriosis. Thus, the aim of the present study was to analyse levels of various VEGFA isoforms in women with and without endometriosis at different stages of the menstrual cycle. Quantitative polymerase chain reaction analysis showed that total VEGFA was highest during menstruation in endometriosis compared with controls (P=0.0373). VEGF121 and VEGF189 were similarly highest during menstruation in endometriosis compared with controls (P=0.0165 and 0.0154 respectively). The present study is also the first to identify the natural expression of VEGF111 in human tissue, which is also highest during menstruation in endometriosis (P=0.0464). This discovery of the natural production of VEGF111 in human endometrium, as well as the upregulation of VEGFA isoforms during menstruation in endometriosis, may shed further light on the development and progression of the disease, and improve our understanding of the regulation of endometrial angiogenesis.


Statistical Analysis of Tract-Tracing Experiments Demonstrates a Dense, Complex Cortical Network in the Mouse.
Anatomical tract tracing methods are the gold standard for estimating the weight of axonal connectivity between a pair of pre-defined brain regions. Large studies, comprising hundreds of experiments, have become feasible by automated methods. However, this comes at the cost of positive-mean noise making it difficult to detect weak connections, which are of particular interest as recent high resolution tract-tracing studies of the macaque have identified many more weak connections, adding up to greater connection density of cortical networks, than previously recognized. We propose a statistical framework that estimates connectivity weights and credibility intervals from multiple tract-tracing experiments. We model the observed signal as a log-normal distribution generated by a combination of tracer fluorescence and positive-mean noise, also accounting for injections into multiple regions. Using anterograde viral tract-tracing data provided by the Allen Institute for Brain Sciences, we estimate the connection density of the mouse intra-hemispheric cortical network to be 73% (95% credibility interval (CI): 71%, 75%); higher than previous estimates (40%). Inter-hemispheric density was estimated to be 59% (95% CI: 54%, 62%). The weakest estimable connections (about 6 orders of magnitude weaker than the strongest connections) are likely to represent only one or a few axons. These extremely weak connections are topologically more random and longer distance than the strongest connections, which are topologically more clustered and shorter distance (spatially clustered). Weak links do not substantially contribute to the global topology of a weighted brain graph, but incrementally increased topological integration of a binary graph. The topology of weak anatomical connections in the mouse brain, rigorously estimable down to the biological limit of a single axon between cortical areas in these data, suggests that they might confer functional advantages for integrative information processing and/or they might represent a stochastic factor in the development of the mouse connectome.


The recovery of touch DNA from RDX-C4 evidences.
RDX (Royal Demolition Explosive) is the organic compound with the formula (O2NNCH2)3. It is a white solid material without smell or taste, widely used as an explosive. It is more energetic explosive than TNT, and it was used widely in World War II. The estimated number of RDX-C4 cases in Bahrain ranged between the years 2015-2018 (May) with a total quantity of 370.72 KG in a total number of 38 cases. The effect of explosive RDX-C4 is very massive and can cause many causalities and fatalities among civilians and policemen. These cases consisted of adhesive film with tapes wrapped around RDX-C4 substance (Demolition Charge M112), black batteries, pipes, black bag contained RDX-C4, and in magnetic improvised explosive device (IED). Touch DNA recovery utilized different collection methods, such as nylon swabbing, tape lifting, and direct cutting of certain parts of the samples that were positive of RDX-C4 through DXR Raman Spectrometer. Samples were extracted and purified with magnetic beads chemistry and quantified. Low copy DNA extracts were subjected to a concentration step. DNA extracts were amplified and processed for detection to obtain reliable results using GlobalFiler Amplification PCR kit and run through ABI 3500xL Genetic Analyzer for fragment length determination. We have discovered that RDX-C4 cannot bind to the DNA nor to the solutions used in DNA typing. Thus, it does not cause DNA inhibition or degradation. From this point of view, we were successful in obtaining acceptable and fit results using advanced techniques. This study will be very useful and informative to assist the forensic community in terrorism case applications worldwide as terrorists do not respect geographical boundaries nor ethnicities of the victims, and the use of DNA profiling technology is the most suitable way to identify the terrorists and keep an end to their violence.


Are we following the WHO recommendations for surgical scrubbing?
Surgical site infections have an incidence as high as 10%. To reduce this, the World Health Organisation (WHO) recommends guidelines for surgical scrubbing. We assessed adherence of surgical staff scrub practice before gowning to current WHO recommendations. A prospective study conducted in operation theatres in a university hospital. Participants comprised of consultant surgeons, trainees and scrub nurses. Variables included staff grade, scrub order, length of scrub time and appropriate dress code. Data were collected anonymously and analysed. We assessed 303 surgical scrub-episodes (95 scrub nurses, 123 trainees and 85 consultant surgeons). Mean time for first scrub of the day was 239 s, SD = 99, 95% CI 218, 259 and range = 530 (scrub nurses = 297 s, trainees = 204 s, consultant surgeons = 202 s). Mean time for subsequent scrubs was 161 s, SD = 63, 95% CI 152, 170, range = 312 (scrub nurses = 184 s, trainees = 158 s, consultant surgeons = 143 s). Statistical significance was found between staff grades based on scrub time, with WHO breaches in remaining variables. No written local guidelines on hand scrubbing exist in the hospital and staff were unaware of WHO guidelines. In this study, mean scrub times were less than WHO guidelines with significant variation in practice across grades of staff. Clear written guidelines are needed.


Molecular dynamics flexible fitting: a practical guide to combine cryo-electron microscopy and X-ray crystallography.
Hybrid computational methods for combining structural data from different sources and resolutions are becoming an essential part of structural biology, especially as the field moves toward the study of large macromolecular assemblies. We have developed the molecular dynamics flexible fitting (MDFF) method for combining high-resolution atomic structures with cryo-electron microscopy (cryo-EM) maps, that results in atomic models representing the conformational state captured by cryo-EM. The method has been applied successfully to the ribosome, a ribonucleoprotein complex responsible for protein synthesis. MDFF involves a molecular dynamics simulation in which a guiding potential, based on the cryo-EM map, is added to the standard force field. Forces proportional to the gradient of the density map guide an atomic structure, available from X-ray crystallography, into high-density regions of a cryo-EM map. In this paper we describe the necessary steps to set up, run, and analyze MDFF simulations and the software packages that implement the corresponding functionalities.


The Strengths and Limitations of South Africa's Search for Apartheid-Era Missing Persons.
This article examines efforts to account for missing persons from the apartheid era in South Africa by family members, civil society organizations and the current government's Missing Persons Task Team, which emerged out of the Truth and Reconciliation Commission process. It focuses on how missing persons have been officially defined in the South African context and the extent to which the South African government is able to address the current needs and desires of relatives of the missing. I make two main arguments: that family members ought to have an active role in shaping the initiatives and institutions that seek to resolve the fate of missing people, and that the South African government ought to take a more holistic 'grave-to-grave' approach to the process of identifying, returning and reburying the remains of the missing.


Detection of sperm surface related antibodies by indirect immunofluorescence test on sperm suspensions. Indirect IFT on sperm suspensions.
An indirect immunofluorescence test (IIFT) using sperm suspensions was carried out on 20 sera with sperm agglutinins (SA) and on 25 negative controls. IIFT gave results highly correlated with the occurrence of sperm agglutinating activity. Moreover, a relation was found among class of Ig involved in IF reactivity, fluorescent stain pattern and type of sperm agglutinations. In all sera with "mixed" or "tail-tail" sperm agglutinating activity, IgG were involved in IF reactivity; the fluorescent stain constantly appeared in a granular pattern along the sperm tail and most often on the head surface too. In relation to high titres of "head-head" sperm agglutinating activity, IgM were involved in IF reactivity; here the fluorescent stain appeared to be localized on acrosomal surface. The results indicate that indirect IFT on sperm suspensions specifically detects sperm surface related antibodies.


Loading of the contralateral foot in peripheral vascular insufficiency below-knee amputees.
Seven ambulatory peripheral vascular insufficiency below-the-knee amputees underwent gait analysis to determine the local pressures on seven prominent plantar regions in their remaining limb. Patients occupied significantly less stance phase time on their remaining limbs. Peak, mean, and total pressures measured at each of the seven prominent areas was decreased as compared with laboratory age and sex matched controls. The results of this study suggest that the risk of contralateral limb amputation surgery is related to systemic peripheral vascular insufficiency or local factors, and not increased loading of the remaining contralateral "limb-at-risk."


Characterization of phenolic and other polar compounds in a lemon verbena extract by capillary electrophoresis-electrospray ionization-mass spectrometry.
In this study, a CE-MS method has been developed to carry out the qualitative characterization of a lemon verbena (Lippia citriodora) extract for the first time. The CE and ESI-MS parameters were optimized with respect to resolution, analysis time and peak shape in order to maximize the number of compounds detected and the sensitivity of their determination. The use of two different MS analyzers, TOF-MS and IT-MS, enabled the tentative identification of the major components of this extract. Thus, using this method, 16 compounds were determined. Some of them have been previously identified by HPLC methods, although four compounds were also found for the first time in lemon verbena such as asperuloside, tuberonic acid glucoside or 5'-hydroxyjasmonic acid 5'-O-glucoside, shanzhiside and ixoside. These results demonstrate that CE-MS generates data complementary to those obtainable by HPLC-MS and it is particularly suited to the analysis of plant metabolites.


Uncovering the roles of PINK1 and parkin in mitophagy.
Parkinson disease (PD) is the second most prevalent neurodegenerative disorder, and thus elucidation of the pathogenic mechanism and establishment of a fundamental cure is essential in terms of public welfare. Fortunately, our understanding of the pathogenesis of two types of recessive familial PDs--early-onset familial PD caused by dysfunction of the PTEN induced putative kinase 1 (PINK1) gene and autosomal recessive juvenile Parkinsonism (ARJP) caused by a mutation in the Parkin gene--has evolved and continues to expand.


A physiological manoeuvre to improve the positivity of the gastro-oesophageal reflux scintigraphic test.
One of the best examinations used routinely to detect gastro-oesophageal reflux (GOR) is scintigraphy and attempts are continuously being made to improve the performance of the test. We have tested a new manoeuvre to improve the results. Three hundred and twenty eight patients (190 males and 138 females, aged 1 month to 84 years, average 4.47 years) were studied after a 12 h fast period with 100 MBq of 99Tcm-phytate, 50 ml volume, directly delivered into the stomach in a mixture of orange juice. All patients were strongly suspected of having GOR. Conventional acquisition was done for 20 min, at a rate of one frame every 20 s. In the middle of the examination, small children were put in the upright position and held there for a few seconds or released in a way that allowed movement; adults were asked to stand up and walk a few metres. The test was then continued for the remaining 10 min. The global index of positivity was 64.6% (76 positive cases in the first half of the examination and 134 positive cases only after the manoeuvre). The mean of GOR episodes observed in the positive cases without the manoeuvre was 0.5 per patient, significantly different from the mean of 1.59 noted after the manoeuvre (P<0.01). In 28 cases the manoeuvre increased either the intensity or the frequency of reflux previously detected without it. We believe that this manoeuvre should be performed in all scintigraphic tests aiming to detect GOR. In the present series, the manoeuvre increased the frequency of GOR episodes.


Chronic disease self-management program for Chinese patients: a preliminary multi-baseline study.
This study reports the preliminary findings on the effects of the Chronic Disease Self-management Program on a group of Chinese participants who suffered from chronic diseases. A total of 23 participants were recruited in a multi-baseline study protocol. Their self-management behaviors, self-efficacy and health status were captured over three baseline assessments and one post-test assessment. The results indicated significant increases in the performance of stretching exercises, the management of cognitive symptoms and communication with physicians. Their self-efficacy in terms of these aspects was found to be significantly increased. However, changes in other aspects of self-management which required more special skills and coordination with outside agencies were not significant. The changes in the physical and mental statuses of the participants were also not significant. It was observed that the positive effects of the program could be attributed to traditional Chinese beliefs of 'self-discipline' and a welcoming response towards self-efficacy strategies. Further studies should adhere to standards of a randomized clinical trial and further examine the mechanisms underpinning the changes in self-management behaviors among Chinese people with chronic diseases.


Learning Outcomes with Linked Assessments - an Essential Part of our Regular Teaching Practice.
Setting up learning outcomes with linked assessments is a best practice in science education. In biology teaching, faculty are beginning to establish learning outcomes and assessments in the style of concept inventories. At a recent meeting of biology faculty who have designed concept inventories, the characteristics and uses of concept inventories were defined. Concept inventories used as pre- and post-measures of student learning provide a window into students' understanding of key concepts of a discipline and serve as a tool to motivate faculty toward evidence-based teaching habits. A movement for the development of a microbiology concept inventory is suggested.


Regulation of vascular endothelial growth factor receptor-2 expression in pancreatic cancer cells by Sp proteins.
Vascular endothelial growth factor receptor-2 (VEGFR2/KDR) is an important mediator of angiogenesis, and VEGFR2 mRNA is expressed in several pancreatic cancer cell lines. Deletion analysis of the VEGFR2 promoter in Panc-1, AsPC-1, and MiaPaCa-2 pancreatic cancer cells shows that the proximal region of the promoter is primarily responsible for VEGFR2 expression, and two GC-rich sites at -58 and -44 are critical elements in all three cell lines. Panc-1, AsPC-1, and MiaPaCa-2 cells also express Sp1, Sp3, and Sp4 proteins which bind to the GC-rich region of the VEGFR2 promoter in electrophoretic mobility shift and chromatin immunoprecipitation assays. RNA interference with small inhibitory RNAs for Sp1, Sp3, and Sp4 decreases VEGFR2 mRNA and reporter gene activity in transfection assays, confirming that VEGFR2 expression in pancreatic cancer cells is regulated by Sp proteins. These results suggest that VEGFR2 cannot only be targeted by receptor tyrosine kinase inhibitors but also by drugs that downregulate Sp proteins or block Sp-dependent transactivation.


Duration of premeiotic deoxyribonucleic acid synthesis and the stages of prophase I in rabbit oocytes.
To estimate the duration of oocyte DNA synthesis 36, 3-day-old female rabbits received 3, 6, 9, 12, 15, or 18 injections of tritiated thymidine (thy-(3)H) at hourly intervals. The ovaries, removed at 1, 10, or 20 days after the first injection, were radioautographed. Counts made of the number of silver grains associated with oocyte nuclei in meiotic Prophase I indicate that the duration of DNA synthesis is between 9 and 12 hr. To determine the length of the stages of meiotic Prophase I, a group of 2-3-day-old rabbits was given a single sub-cutaneous injection of thy-(3)H, and the ovaries were removed at hourly and/or daily intervals after treatment. The minimum duration of leptotene was 3 hr and the maximum duration probably was less than 8 hr. The maximum durations of zygotene, pachytene, and diplotene were estimated to be 44, 216, and 96 hr, respectively. The interval from the end of oogonial DNA synthesis to the beginning ofpremeiotic DNA synthesis (G(2) + Mitosis + G(1)) appeared to be less than 6 hr.


Fracture Resistance and Cusp Deflection of Lined or Non-lined Composite and Glass Hybrid Restorations Over Residual Demineralized Dentin.
To assess the fracture resistance (FR) and cusp deflection (CD) of lined or non-lined composite (CO) and glass hybrid (GH) restorations over residual demineralized dentin. In 48 extracted human premolars, artificial residual demineralized dentin was induced on pulpo-axial walls of standardized cavities. Various restorations were placed over this demineralized dentin: an experimental GH, a composite restoration (OptiBond FL+Tetric EvoCeram) without lining, or composite restorations with non-setting (Hypocal) or setting (Dycal) calcium hydroxide lining. After thermomechanical cycling, groups (n = 12) were compared regarding their CD and FR. CD did not differ significantly between groups. FR was significantly lower in teeth restored with GH (median: 238 N; 25th/75th percentiles: 191/287 N) than in those restored with lined or non-lined composites (median range: 517-569 N; p < 0.05/Mann-Whitney), which did not differ significantly from each other (p > 0.05). Within the conditions of this in vitro study, CH lining of pulpo-axial walls had only limited impact on CD and FR. GH showed the lowest FR and might not be optimal for restoring deep or extended cavitated lesions.


Embryonic stem cell research and therapy: the need for a common European legal framework.
The possibility of obtaining stem cells from human embryos has given rise to an intensive legal and ethical debate. In this paper, attention is paid to the normative disparity and ambiguity in Europe. An argument for the need for a minimum legal harmonization is made; and a prudent and flexible way to reach this successfully is suggested. Establishing a common legal framework seems to be the only way to guarantee true competitiveness for the European scientific community.


Increased secretion of insulin-like growth factor-binding proteins and decreased secretion of insulin-like growth factor-II by muscle from growth-retarded neonatal rats.
Insulin-like growth factors (IGFs) and IGF-binding proteins (IGFBPs) may be important factors in the control of neonatal growth. We have examined the production, in vitro, of IGFBPs and IGFs by hind-limb skeletal muscle from normal and small-for-gestational age (SGA) neonatal rats. Conditioned medium was collected from muscle strips after incubation at 37 degrees C for 2 h in Ham's F-12 medium. The conditioned medium was subjected to acid-gel permeation chromatography to separate IGFBPs from IGFs. The binding of 125I-labelled IGF-I to IGFBPs from both control and SGA muscle was displaced equipotently by IGF-I and IGF-II and not at all by insulin. IGFBPs from control and SGA muscles bound IGF-I with comparable affinities (Kd = 0.071 and 0.069 nmol/l respectively). When IGF-II was used as tracer, neither IGF-I nor insulin competed for binding. Western ligand blots of IGFBPs in conditioned media from both control and SGA muscles showed three bands of radioactivity at molecular masses equivalent to 24, 30 and 40 kDa. When the release of IGFBPs by muscle tissue in vitro was quantified by measuring the number of IGF-I binding sites in acid-fractionated medium it was apparent that the muscles from SGA pups secreted significantly more IGFBPs (39.3 +/- 7.5 fmol/mg muscle protein per 2 h) than the muscles from control pups (17.8 +/- 2.7 fmol/mg protein per 2 h; P less than 0.05). In contrast to the IGFBPs, more IGF activity was secreted by the muscles from the control pups (61.1 +/- 15.6 fmol/mg muscle protein per 2 h) than the muscles from the SGA pups (12.6 +/- 5.8 fmol/mg muscle protein per 2 h; P less than 0.05). Analysis of the IGF activity with assays specific for IGF-I and IGF-II showed that both SGA and control muscles secreted predominantly IGF-II with approximately 10% of the total IGF activity measurable as IGF-I. This differential secretion of IGFBPs and IGFs may be associated with the reduced growth potential of the SGA neonate.


Tunable Er-doped fiber ring laser with single longitudinal mode operation based on Rayleigh backscattering in single mode fiber.
A tunable and single longitudinal mode Er-doped fiber ring laser (SLM-EDFRL) is proposed and demonstrated based on Rayleigh backscattering (RBS) in single mode fiber-28e (SMF-28e). Theory and experimental study on formation of SLM from normal multi-mode ring laser is demonstrated. The RBS feedback in 660 m SMF-28e is the key to ensure SLM laser oscillation. This tunable SLM laser can be tuned over 1549.7-1550.18 nm with a linewidth of 2.5-3.0 kHz and a side mode suppression ratio (SMSR) of ~72 dB for electrical signal power. The tuning range is determined by the bandpass filter and gain medium used in the experiment. The laser is able to operate at S+C+L band.


Empirical antifungal therapy for patients with neutropenia and persistent fever: Systematic review and meta-analysis.
To assess the evidence for the current standard of practice of using empirical antifungal treatment in febrile neutropenic cancer patients. Systematic review and meta-analysis of randomised controlled trials comparing empirical or preemptive antifungal treatment with placebo, no intervention, or another antifungal. The primary outcomes were all-cause mortality and invasive fungal infections (IFI) (documented or probable). Relative risks (RR) with 95% confidence intervals (CI) were pooled. Six trials assessed the efficacy of empirical treatment compared to no treatment and one compared empirical to preemptive therapy. Empirical treatment did not decrease mortality significantly (RR 0.82, 95% CI 0.50-1.34), but significantly decreased IFIs (RR 0.25, 0.12-0.54). Twenty-three trials assessed the efficiency of different antifungals. All-cause mortality was lower with azoles compared to amphotericin B (AB) (RR 0.81, 0.65-1.01); IFI rates were not different while adverse events were less frequent with azoles (RR 0.40; 0.34-0.66). Liposomal AB was associated with lower mortality and IFIs than other AB formulations (RR 1.57, 1.10-2.23 and 1.48, 0.98-2.25, respectively). Caspofungin was associated with fewer adverse events, but otherwise comparable to liposomal AB. All trials included patients with haematological malignancies. Major limitations included per-protocol analysis, non-blinded design and inconsistent definitions of IFIs. Empirical antifungal treatment is associated with a lower rate of IFIs but no significant difference in overall mortality. The assessment of IFIs in these trials may have been biased, offering only weak support to standard practice. Azoles, liposomal amphotericin B or caspofungin should be preferred. Pre-emptive antifungal therapy should be considered and further investigated.


Sickle erythrocytes adhere to polymorphonuclear neutrophils and activate the neutrophil respiratory burst.
The vasoocclusive process in patients with sickle cell disease (SCD) is complex and involves interactions among sickle erythrocytes (SS-RBC), vascular endothelium, and plasma and cellular components. The role of neutrophils (PMN) in vasoocclusion has not been examined. Patients with SCD appear to have chronically activated PMN. Because the first step in PMN activation is particle recognition, we explored whether normal PMN recognize SS-RBC and whether this recognition results in PMN monolayers, significantly more SS-RBC adhered to the PMN than did normal erythrocytes (AA-RBC; P < .001). Preincubation of erythrocytes with autologous plasma significantly increased the adherence of SS-RBC to PMN but had no effect on AA-RBC (P < .001). When adhesion of density fractionated SS-RBC was performed, dense SS-RBC showed greater adherence to the PMN monolayers than did light SS-RBC (P < .001). To determine mechanisms of this adhesion, IgG and Arg-Gly-Asp-Ser (RGDS) receptor sites on PMN were saturated. IgG inhibited adherence of dense SS-RBC, whereas RGDS inhibited adherence in both fractions, although to a greater extent in the light fraction. We measured SS-RBC activation of PMN by incubating SS-RBC with 2', 7'-Dichloro-fluroescin Diacetate (DCF)-labeled PMN. Incubation of PMN with SS-RBC resulted in a significant increase in fluorescence compared to AA-RBC. We show here that PMN recognize SS-RBC through multiple mechanisms and that this recognition results in activation of PMN. These findings contribute to the understanding of vasoocclusive crisis in patients with SCD and may have therapeutic implications.


Photothermal Spectro-Microscopy as Benchmark for Optoplasmonic Bio-Detection Assays.
Optoplasmonic bio-detection assays commonly probe the response of plasmonic nanostructures to changes in their dielectric environment. The accurate detection of nanoscale entities such as virus particles, micelles and proteins requires optimization of multiple experimental parameters. Performing such optimization directly via analyte recognition is often not desirable or feasible, especially if the nanostructures exhibit limited numbers of analyte binding sites and if binding is irreversible. Here we introduce photothermal spectro-microscopy as a benchmarking tool for the characterization and optimization of optoplasmonic detection assays.


Gene expression analysis in the larval silk gland of the eri silkworm Samia ricini.
Insects produce silk for a range of purposes. In the Lepidoptera, silk is utilized as a material for cocoon production and serves to protect larvae from adverse environmental conditions or predators. Species in the Saturniidae family produce an especially wide variety of cocoons, for example, large, golden colored cocoons and those with many small holes. Although gene expression in the silk gland of the domestic silkworm (Bombyx mori L.) has been extensively studied, considerably fewer investigations have focused on members of the saturniid family. Here, we established expression sequence tags from the silk gland of the eri silkworm (Samia ricini), a saturniid species, and used these to analyze gene expression. Although we identified the fibroin heavy chain gene in the established library, genes for other major silk proteins, such as fibroin light chain and fibrohexamerin, were absent. This finding is consistent with previous reports that these latter proteins are lacking in saturniid silk. Recently, a series of fibrohexamerin-like genes were identified in the Bombyx genome. We used this information to conduct a detailed analysis of the library established here. This analysis identified putative homologues of these genes. We also found several genes encoding small silk protein molecules that are also present in the silk of other Lepidoptera. Gene expression patterns were compared between eri and domestic silkworm, and both conserved and nonconserved expression patterns were identified for the tested genes. Such differential gene expression might be one of the major causes of the differences in silk properties between these species. We believe that our study can be of value as a basic catalogue for silk gland gene expression, which will yield to the further understanding of silk evolution.


Ectopic pregnancy: common and some uncommon misdiagnoses.
Several diseases of the peritoneal cavity may present in manners that are difficult to distinguish from ectopic pregnancy. A careful history and a thorough physical examination are paramount to making the correct diagnosis. Diseases of the gastrointestinal and urinary tracts usually present in specific manner. The difficulty arises when the clinical presentations of these disease processes are atypical. Under such circumstances, other diagnostic modalities, including serial beta-hCG testing and pelvic ultrasound, are extremely useful in distinguishing ectopic pregnancy from other diseases that occur in the abdominal peritoneal cavity. Generous use of the laparoscope facilitates diagnostic exclusion of certain entities, decreases the frequency of misdiagnosis, prevents unnecessary surgical procedures, and reduces morbidity and mortality.


COVID-19 in Human, Animal, and Environment: A Review.
The medical authority in China, especially in Wuhan city, reported on December 2019 a large number of highly fatal, rapidly spreading viral pneumonia caused by an unknown coronavirus. The common history of all the patients was their visiting a Wuhan's whole food store, where live animals and seafood are sold. Irrespective of the efforts of the Chinese authorities, the virus spread rapidly all over the world by travelers, provoking widespread attention by the media and panic. Many previous coronavirus epidemics had been recorded, such as severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS), and the recently newly discovered epidemic is named coronavirus disease of 2019 (COVID-19). This disease is caused by SARS Coronavirus-2 (SARS-CoV-2), and this virus is antigenically related to the SARS virus (SARS-CoV), which had been detected in 2002, depending on clinical, serological, and molecular findings. There is rapid competition among the researchers to discover the source of the virus, understand the mechanism of the disease development, establish treatment strategies, and determine the factors affecting the incidence of infection and severity of the disease, and focus on the production of a vaccine. Coronaviruses are a group of single-stranded, positive-sense RNA genome viruses; its genome length varies from 26 to 32 kb. Coronavirus causes mild to severe respiratory disorders. In December 2019, several cases of pneumonia of unknown causes were found in Wuhan city, which is located in the Hubei province in China. Chinese health authorities investigated the problem and found that a new virus caused such infection and, using next-generation sequencing, found the 2019 novel coronavirus (2019-nCoV). It has been transferred from humans to humans and animals to humans (zoonotic). Coronaviruses cause multiple respiratory problems, varying from common cold to severe infections such as SARS. General symptoms of infection include fatigue, cough, and breathing problems such as shortness of breath, as described by World Health Organization. Serious cases may result in pneumonia, renal failure, and even death. We address current information about the new SARS Coronavirus-2 as well as the COVID-19 disease caused by it in this review.


Dowry Prohibition (Maintenance of Lists of Presents to the Bride and Bridegroom) Rules, 1985, 11 August 1985.
These Rules are made under Section 9 of the Dowry Prohibition Act, 1961, of India and provide, among other things, that the list of presents given to either the bride or bridegroom is to be prepared as soon as possible after the marriage; is to be in writing; is to contain a brief description of each gift, including value, name of the person presenting it, and how the person presenting it is related to either spouse; and is to be signed by both spouses.


Experimental infection of turbot, Scophthalmus maximus (L.), by Moritella viscosa, vaccination effort and vaccine-induced side-effects.
Moritella viscosa is the causative agent of winter ulcers in farmed salmonids and Atlantic cod in countries around the North Atlantic. The bacterium has also been isolated from various marine fish species. Bacterial diseases have been a limiting factor in farming of turbot, but M. viscosa has not so far been isolated. In this study, turbot was shown to be sensitive to M. viscosa infection in experimental challenges. Pathological changes in infected turbot were comparable with those previously described for winter ulcers in salmon. A multivalent commercial salmon vaccine, containing M. viscosa as one of five antigens and a mineral oil adjuvant, did not protect turbot against challenge 13 weeks post-vaccination. Weight gain of vaccinated turbot compared with controls was not reduced 7 weeks post-vaccination. Vaccination did not induce a specific anti-M. viscosa response, while elevated anti-M. viscosa antibody levels were detected both in vaccinated and unvaccinated fish 5 weeks post-challenge. The vaccine did, however, induce an antibody response against Aeromonas salmonicida, another vaccine component. Minor intra-abdominal adhesions were detected in vaccinated fish and fish injected with a mineral oil adjuvant. The measurement of various innate humoral immune parameters did not reveal significant differences between vaccinated and control groups.


Bilateral orbital infarction and retinal detachment in a previously undiagnosed sickle cell hemoglobinopathy African child.
Bone infarction involving the orbit in sickle cell disease is not common. Bilateral orbital infarction in a previously undiagnosed sickle cell hemoglobinopathy has not been previously reported. In this report, we present a case of an 11-year-old previously undiagnosed sickle cell disease Nigerian girl with severe acute bilateral orbital infarction and retinal detachment to highlight that hemoglobinopathy induced orbital infarction should be considered in African children with acute onset proptosis with or without previous history of sickle cell hemoglobinopathy.


Dietary intake of aluminum in a Spanish population (Canary Islands).
The aim of this study was to analyze the aluminum content in foods and beverages most commonly consumed by the Canary Island population to determine the dietary intake of this metal throughout the Canary Islands as a whole and in each of the seven islands (Gran Canaria, Lanzarote, Fuerteventura, Tenerife, La Palma, La Gomera, and El Hierro). Four hundred and forty samples were analyzed by ICP-OES. Estimated total intake of aluminum for the Canary population was 10.171 mg/day, slightly higher than the provisional tolerable weekly intake (PTWI; 10 mg/day for a person weighing 70 kg). Aluminum intake by age and sex of the Canary Island population was also determined and compared values from other populations, both national and international.


Virus-Receptor Interactions of Glycosylated SARS-CoV-2 Spike and Human ACE2 Receptor.
The SARS-CoV-2 betacoronavirus uses its highly glycosylated trimeric Spike protein to bind to the cell surface receptor angiotensin converting enzyme 2 (ACE2) glycoprotein and facilitate host cell entry. We utilized glycomics-informed glycoproteomics to characterize site-specific microheterogeneity of glycosylation for a recombinant trimer Spike mimetic immunogen and for a soluble version of human ACE2. We combined this information with bioinformatics analyses of natural variants and with existing 3D structures of both glycoproteins to generate molecular dynamics simulations of each glycoprotein both alone and interacting with one another. Our results highlight roles for glycans in sterically masking polypeptide epitopes and directly modulating Spike-ACE2 interactions. Furthermore, our results illustrate the impact of viral evolution and divergence on Spike glycosylation, as well as the influence of natural variants on ACE2 receptor glycosylation. Taken together, these data can facilitate immunogen design to achieve antibody neutralization and inform therapeutic strategies to inhibit viral infection.


Comparison of sleep characteristics during the first and second period of restrictive measures due to COVID-19 pandemic in Greece.
The first wave of coronavirus pandemic and concomitant restrictive measures affected sleep. We slept more than usual, but the quality was worse. As the pandemic continues, a second period of restrictive measures was initiated, and no data exist about their impact on sleep. The aim of this study was to compare quantity and quality of sleep between the two periods of restrictive measures, due to the Coronavirus Disease-19 (COVID-19) pandemic in Greece. A web-based survey using a short 13-item questionnaire was created and was distributed online. This included information about demographic and professional data, quantitative and qualitative characteristics of sleep, degree of abidance in lockdown measures, and data about COVID-19 infection or close contact with active confirmed cases. A total of 1,078 questionnaires were evaluated (first period, n=963; second period, n=115). Sleep duration was shorter during the second lockdown (mean difference -0.51h; 95% confidence interval, (CI), -0.82, -0.19, p=0.002). Compared to usual habits, sleep increased in first wave (mean difference +0.37h; 95% CI, 0.26, 0.47; p<0.001) and decreased in second wave (mean difference -0.35h; 95% CI, -0.60, -0.09; p=0.009). Regarding quality of sleep, less participants reported good quality of sleep during the second wave compared to the first (p=0.006). Finally, compliance to restriction measures was lower and the prevalence of confirmed illness or close contact with COVID-19 cases among participants was higher during the second period than during the first one (p<0.001 and p=0.028, respectively). Our data showed that sleep duration increased during the first lockdown and decreased in the second one, when compared to usual habits. Moreover, sleep quality progressively deteriorated, as the restrictive measures due to coronavirus pandemic continued.