Biological Evaluation of 99mTc-HYNIC-EDDA/tricine-(Ser)-D4 Peptide for Tumor Targeting.
D4 small peptide (Leu-Ala-Arg-Leu-Leu-Thr) was selected as an appropriate agent for specific targeting of epidermal growth factor receptor (EGFR). The aim of study was to investigate the 99mTc-labeled D4 peptide for non-small cell lung tumor targeting. HYNIC-(Ser)3-D4 peptide was labeled with 99mTc using mixture of tricine and ethylenediamine diacetic acid (EDDA) as co-ligands. The in vitro cellular uptake of radiolabeled peptide was evaluated by blocking test on human non-small cell lung cancer (A-549) cell line and its biodistribution was evaluated in A-549 xenografted nude mice. This conjugated peptide was labeled with 99mTc in high radiochemical purity and it was highly stable in buffer and serum. The un-blocked to blocked cellular radioactivity ratio was 4- fold that showed a specific binding of this radiolabeled peptide on A-549 cell. Animal biodistribution in A-549 xenografted nude mice showed rapid clearance from blood and other non-target organs. Tumor uptake values as %ID/g (percentage of injection dose per gram of tissue) were 2.47% and 1.30% at 1 and 4 h after injection. This study showed the 99mTc-EDDA/tricine-HYNIC-(Ser)3-D4 peptide had tumor targeting on the non-small cell lung tumor.


Immunophenotypic signature of primary glioblastoma multiforme: A case of extended progression free survival.
Glioblastoma-multiforme (GBM), the most aggressive glial tumor, has a worldwide age-adjusted incidence ranging from 0.59-3.69/100000 persons. Despite current multimodal-treatment approach, median-survival time and progression-free survival (PFS) remains short. Glioblastomas display a variety of molecular alterations, which necessitates determining which of these have a prognostic significance. This is a case of a 45-year-old patient who presented with progressive slurring of speech and features of raised intracranial pressure. Computed tomography (CT) scan revealed a large heterogeneously enhancing lesion in the left front-temporal-perisylvian region with solid, cystic areas, suggestive of malignant glioma. Partial tumor-excision was followed by concurrent chemo-radiotherapy. Histopathologically, the tumor was astrocytoma grade-IV. Patient had an extended PFS of 12 mo, with an overall survival of 26 mo. Primary-GBM was confirmed using molecular markers and the immunophenotypic signature was defined by evaluating systemic expression of human telomerase reverse transcriptase, interleukin-6, neutrophil-lymphocyte ratio, tissue inhibitor of metalloproteinases-1, human chitinase-3-like-protein-1 (YKL-40) and high mobility group-A1. Current findings suggest that this signature can identify worst outcomes, independent of clinical criteria.


Two organically templated niobium and zinconiobium fluorophosphates: low temperature hydrothermal syntheses of NbOF(PO4)2(C2H10N2)2 and Zn3(NbOF)(PO4)4(C2H10N2)2.
Two new niobium and zinconiobium fluorophosphates, NbOF(PO4)2(C2H10N2)2 (1) and Zn3(NbOF)(PO4)4-(C2H10N2)2 (2), have been prepared under hydrothermal conditions using ethylenediamine as a template. The structures were determined by single crystal diffraction to be triclinic, space group P1 (No. 2), a = 8.1075 (6) A, b = 9.8961 (7) A, c = 10.1420(8) A, alpha = 111.655(1) degrees, beta = 111.51(1) degrees, gamma = 93.206(1) degrees, V = 686.19(9) A3, and Z = 2 for 1 and orthorhombic, space group Fddd (No. 70), a = 9.1928(2) A, b = 14.2090(10) A, c = 32.2971 (6) A, V = 4218.66(12) A3, and Z = 8 for 2, respectively. Compound 1 is an infinite linear chain consisting of corner-sharing [Nb2P2] 4-MRs bridged at the Nb centers with organic amines situated between chains, and compound 2, containing the chains similar to that in 1, forms a zeotype framework with organic amines situated in the gismondine-type [4684] cavities. The topology of 2 was previously unknown with vertex symbol 4 x 4 x 4 x 4 x 8 x 8 (vertex 1), 4 x 4 x 4 x 82 x 8 x 8 (vertex 2), 4 x 4 x 8 x 8 x 82 x 82 (vertex 3), and 4 x 4 x 4 x 82 x 8 x 8 (vertex 4). The topological relationships between the 4-connected network of 2 and several reported (3,4)-connected networks were discussed.


International migration: new challenges to Europe.
This report, which was commissioned by the Secretary General of the Council of Europe, is an overview of the situation concerning current and future trends in international migration in Europe. The most important components of the current scene are identified as "(i) persisting unemployment despite the economic recovery in Europe, (ii) the continued internationalisation of the national societies in conjunction with overt tendencies to xenophobia, (iii) the strengthening of European political and economic integration, and (iv) the increasing pressure for immigration from the third world."


Utilizing 3-Dimensional Cardiac Models With Point-of-Care Ultrasound Video Tutorials to Improve Medical Student Education: A Double-Blinded Randomized Control Study.
Introduction Ultrasound is a rapidly expanding imaging modality that many medical schools are incorporating into a structured curriculum. Learning both anatomy and ultrasound imaging simultaneously is intuitively challenging. This double-blinded, randomized control study examined the effect of utilizing three-dimensional (3d) cardiac models within an ultrasound video tutorial in order to achieve improved cardiac ultrasound anatomy education. Methods Thirty-nine (39) first- and second-year medical students at a single medical school voluntarily participated. The control group watched a video tutorial on cardiac ultrasound anatomy while the experimental group watched a similar video tutorial that also included a 3d cardiac model. The effect was measured with a multiple-choice test that included a sub-analysis of ultrasound principles. The test was unique in that no text or context clues were provided on the reference images, further challenging anatomic identification. Results The findings of the study included a p-value of 0.73 for the ultrasound principles section and a p-value of 0.77 for the cardiac anatomy. There was no statistical difference in the primary outcome or in the subgroup analysis. Post-hoc analysis demonstrated the study was underpowered. Conclusions This study is the first of its kind to utilize an innovative testing method that holds promise for future research in regards to utilizing 3d models with ultrasound education. The study was underpowered, therefore no definitive conclusions about the utility of 3d cardiac models in the educational process can be ascertained.


Methyl 3-(4-chloro-phen-yl)-2-(1,3-dimethyl-2,5-dioxo-4-phenyl-imidazolidin-4-yl)-3-oxopropano-ate.
The title compound, C(21)H(19)ClN(2)O(5), is a tetra-substituted hydantoin derivative which contains an imidazolidine-2,4-dione core. The dihedral angle between the aromatic rings is 64.53 (14)°. In the crystal, weak inter-molecular C-H⋯O hydrogen bonding is found. An intra-molecular C-H⋯O inter-action also occurs.


Electrospun/3D-Printed Bicomponent Scaffold Co-Loaded with a Prodrug and a Drug with Antibacterial and Immunomodulatory Properties.
This work reports the construction of a bicomponent scaffold co-loaded with both a prodrug and a drug (BiFp@Ht) as an efficient platform for wound dressing, by combining the electrospinning and 3D-printing technologies. The outer component consisted of a chitosan/polyethylene oxide-electrospun membrane loaded with the indomethacin-polyethylene glycol-indomethacin prodrug (Fp) and served as a support for printing the inner component, a gelatin methacryloyl/sodium alginate hydrogel loaded with tetracycline hydrochloride (Ht). The different architectural characteristics of the electrospun and 3D-printed layers were very well highlighted in a morphological analysis performed by Scanning Electron Microscopy (SEM). In vitro release profile studies demonstrated that both Fp and Ht layers were capable to release the loaded therapeutics in a controlled and sustained manner. According to a quantitative in vitro biological assessment, the bicomponent BiFp@Ht scaffold showed a good biocompatibility and no cytotoxic effect on HeLa cell cultures, while the highest proliferation level was noted in the case of HeLa cells seeded onto an Fp nanofibrous membrane. Furthermore, the BiFp@Ht scaffold presented an excellent antimicrobial activity against the E. coli and S. aureus bacterial strains, along with promising anti-inflammatory and proangiogenic activities, proving its potential to be used for wound dressing.


Standardized Subannular Repair Improves Outcomes in Type IIIb Functional Mitral Regurgitation.
The major drawback of isolated annuloplasty in secondary mitral regurgitation (MR) is the reoccurrence of MR. We prospectively compared the results of isolated annuloplasty vs annuloplasty with simultaneous standardized subannular repair. The study comprised 101 patients with secondary type IIIb MR. Of these, 51 underwent annuloplasty plus standardized subannular repair with realignment of both papillary muscles (subannular repair) and 50 underwent isolated annuloplasty. The primary study end point was the reoccurrence of MR >2 at the 1-year follow-up. Secondary end points were survival, freedom from major adverse cardiac events, and residual leaflet tethering. Baseline characteristics were comparable in both groups. There was no significant difference in in-hospital mortality (P = .3). Although postrepair MR was comparable between the subannular repair and isolated annuloplasty subgroups, the residual leaflet tethering (tenting area, 127.6 ± 35.8 mm2 vs 166.3 ± 47.3 mm2, P = .02; posterior mitral leaflet angle, 19.2 ± 4.7 degrees vs 24.8 ± 5.2 degrees, P = .001; anterior mitral leaflet angle, 25.4 ± 5.8 degrees vs 34.1 ± 4.0 degrees, P = .001; and tenting height, 5.9 ± 1.4 mm vs 9.2 ± 2.2 mm, P = .001) were significantly increased in the isolated annuloplasty group (P < .001). At the 1-year follow-up, we found a significant difference between the groups in the freedom from MR >2 of 98% (50 of 51) for subannular repair vs 86.7% (39 of 45) for isolated annuloplasty (P = .045) and mortality of 0% (0 of 51) for subannular repair vs 10% (5 of 50) for isolated annuloplasty (P = .025). In secondary MR with reduced leaflet motion, the combination of annuloplasty and standardized subannular repair is associated with a significantly reduced MR reoccurrence, decreased residual leaflet tenting, and significantly improved 1-year outcome compared with annuloplasty alone.


Organization development in a psychiatric hospital: creating desirable changes.
The organization of the way in which hospitals and hospital staff provide a service to patients is obviously of critical importance to their effectiveness, yet it is clear that rigidities and inappropriate and ineffective procedures frequently intrude. It is commonly held that changing hospitals as organizations is difficult to acomplish, and indeed, reported attempts at such change reflect this. The project reported here was a successful attempt at changing a number of different aspects of the culture of a psychiatric hospital which included managerial practices and structure, aspects of patient care, multidisciplinary team work, and staff development. The present paper concentrates on some specific outcomes at ward level. The general pattern for bringing about change involves the collection of (valid) data and then feeding this back to the staff involved so that they can take appropriate action. The data discussed here concerned ward nursing staff's attitude to the 'climate' of the hospital, their job satisfaction and aspects of patient care. This was fed back to nursing, managerial and medical staff, and action plans were agreed to overcome the difficulties highlighted. Outcomes have included the production of ward and unit objectives and changes in treatment programmes and aspects of patient care on the wards.


Suspensory ligament release combined with acellular dermal matrix filler in infrapubic space: A new method for penile length augmentation.
Until now, no reliable method is recognised in treating buried penis. This study explored a new method of penile length augmentation using acellular dermal matrix filler in infrapubic space combined with liposuction and penile suspensory release. Patients with "small-sized penis" received penile length augmentation procedure including suprapubic liposuction, penile suspensory ligament release and insertion of folded acellular dermal matrix between corpora cavernosa and pubis symphysis. Their penile length from tip to skin was measured pre-operatively and post-operatively. The post-operative complications and patients' satisfaction were also recorded. Fifteen adult male patients were included with the mean age of 33.2 ± 4.6 years old and BMI of 28.9 ± 5.3 kg/m2 . The average amount of liposuction was 430 ± 90.0 ml. The average penile length measured pre-operatively and post-operatively (on table and 3 months afterwards) was 3.0 ± 1.3 cm, 7.3 ± 2.1 cm and 5.4 ± 1.8 cm. The penile length has significantly increased by 4.3 ± 1.6 cm (on table) and 2.4 ± 0.8 cm (3 months post-operatively; p < 0.05). The post-operative complications included oedema of penis, ecchymosis of lower abdomen and poor wound healing. No patient was dissatisfied with the appearance and function. The new method using acellular dermal matrix combined with liposuction and penile suspensory ligament release is safe and effective. The method could be applied to selected patients with buried penis.


[Rhabdomyolysis, disseminated intravascular coagulation and acute renal failure after severe narcotics intoxication (MDMA, THC, amphetamine)].
More often we are faced with the cases of young people (who are in a serious condition) who land in ICU, because of severe narcotics intoxication, which they took occasionally on the concerts, discotheques and social events. From 1997 we observed rapid increase of admission to hospitals due to amphetamine, MDMA (2,3-methylenedeoxymethamphetamine which is a main component of a tablet called ecstasy) and THC (9-d tetrahydrocannabinols which are a component of sunn hemps) intoxication and decrease of opioid's poisoning. 23 years old patient was admitted to ICU in critical condition after severe narcotics intoxication. Patient was deeply unconscious (GCS 3) with tetraplegia and high temperature (39.6 degrees C). He had endotracheal tube (artificially ventilated) and hypovolemic shock with circulatory insufficiency (blood pressure was supported by 3 catecholamines). We observed many petechias and ecchymoses which suggested vascular haemorrhagic diathesis. It was found that the patient had disseminated intravascular coagulation and rhabdomyolysis with acute renal failure which was treated by dialysis. After 26 days of intensive treatment the patient was conscious, he had also efficient circulatory and respiratory system but with slight improvement of neurological state. In this condition he was admitted on nephrology ward to continue the treatment and start rehabilitation. The presence of high concentration of amphetamine, MDMA and THC in blood, extreme dehydration and electrolytes disturbances caused rhabdomyolysis, DIC syndrome and acute renal failure which make the prognosis worse and complicate the treatment. Estimating probability of death of this patient in SAPS II scale (Simplified Acute Physiology Scale) he has bad prognosis (86 points gives 95% of death probability). The patient is alive (what is a big success), probably thanks to quick arrival to specialized medical centre and dialysis treatment which was started early.


Oral Versus Intragastric Inoculation: Similar Pathways of Trypanosoma cruzi Experimental Infection? From Target Tissues, Parasite Evasion, and Immune Response.
Currently, oral infection is the most frequent transmission mechanism of Chagas disease in Brazil and others Latin American countries. This transmission pathway presents increased mortality rate in the first 2 weeks, which is higher than the calculated mortality after the biting of infected insect vectors. Thus, the oral route of Trypanosoma cruzi infection, and the consequences in the host must be taken into account when thinking on the mechanisms underlying the natural history of the disease. Distinct routes of parasite entry may differentially affect immune circuits, stimulating regional immune responses that impact on the overall profile of the host protective immunity. Experimental studies related to oral infection usually comprise inoculation in the mouth (oral infection, OI) or gavage (gastrointestinal infection, GI), being often considered as similar routes of infection. Hence, establishing a relationship between the inoculation site (OI or GI) with disease progression and the mounting of T. cruzi-specific regional immune responses is an important issue to be considered. Here, we provide a discussion on studies performed in OI and GI in experimental models of acute infections, including T. cruzi infection.


Welfare reform: loss of supplemental security income (SSI) for children with disabilities.
To describe the changes the new welfare reform legislation, the Personal Responsibility and Work Opportunity Reconciliation Act of 1996, makes in children's eligibility for Supplemental Security Income (SSI) and the implications of the changes for low-income children with disabilities and their families. Low-income children and youth, 18 years of age and younger, who have one or more limitations in physical or mental function in performing age-appropriate skills. The tightened federal standards for determining whether children are disabled place thousands of children at risk for losing SSI cash benefits. This loss of necessary financial support has the potential to adversely affect the physical and psychological health of low-income children with disabilities and their families. The new law represents a step backward from the long awaited gains the 1990 Zebley v. Sullivan U. S. Supreme Court decision legislated for children with disabilities. Knowledge of the legal changes in children's SSI enables nurses to implement interventions that help low-income families maintain SSI eligibility for their children with disabilities and to advocate with the public and policymakers on behalf of this vulnerable population.


Kinetically Controlled Self-Assembly of Phosphorescent AuIII Aggregates and Ligand-to-Metal-Metal Charge Transfer Excited State: A Combined Spectroscopic and DFT/TDDFT Study.
Metallophilic interactions in d10-d10(AuI-AuI)/d8-d8(PtII-PtII, RhI-RhI, IrI-IrI) complexes have been widely studied for decades, and metal-metal (M-M) bonding character has been revealed in both the ground and excited states. These M-M closed-shell interactions are appealing driving forces for the self-assembly of supramolecular/polymeric systems, providing luminescent properties distinctly different from those of the corresponding monomer. However, reports on attractive interactions between two AuIII complex cations are scarce in the literature. Herein is described a series of pincer-type cationic AuIII complexes with different auxiliary ligands, among which the AuIII-allenylidene complex displays a close Au-Au contact of 3.367 Å between neighboring molecules in its X-ray crystal structure; AuIII-isocyanide complexes show a broad red-shifted absorption band and prominent phosphorescence upon aggregation that was influenced by an attractive AuIII-AuIII bonding interaction in the excited state; and AuIII-acetylene complexes can undergo living supramolecular polymerization upon varying the counteranion. The nature of the emissive excited state(s) of the AuIII aggregates is assigned to a mixture of major 3[π-π*] and minor 3LMMCT (ligand-to-metal-metal charge transfer) states based on combined spectroscopic and DFT/TDDFT studies. The morphology of the AuIII aggregates is highly dependent on the concentration and nature of the counteranion. A qualitative model has been applied to account for the concentration- and counteranion-dependent kinetics of the supramolecular polymerization process.


Identification of novel mutations of the WFS1 gene in Brazilian patients with Wolfram syndrome.
Wolfram syndrome (WS) is a rare, progressive, neurodegenerative disorder with an autosomal recessive pattern of inheritance. The gene for WS, WFS1, was identified on chromosome 4p16 and most WS patients carry mutations in this gene. However, some studies have provided evidence for genetic heterogeneity and the genotype-phenotype relationships are not clear. Our aim was to ascertain the spectrum of WFS1 mutations in Brazilian patients with WS and to examine the phenotype-genotype relationships in these patients. Clinical characterization and analyses of the WFS1 gene were performed in 27 Brazilian patients with WS from 19 families. We identified 15 different mutations in the WFS1 gene in 26 patients, among which nine are novel. All mutations occurred in exon 8, except for one missense mutation which was located in exon 5. Although we did not find any clear phenotype-genotype relationship in patients with mutations in exon 8, the homozygous missense mutation in exon 5 was associated with a mild phenotype: onset of diabetes mellitus and optic atrophy during adulthood with good metabolic control being achieved with low doses of sulfonylurea. Our data show that WFS1 is the major gene involved in WS in Brazilian patients and most mutations are concentrated in exon 8. Also, our study increases the spectrum of WFS1 mutations. Although no clear phenotype-genotype relationship was found for mutations in exon 8, a mild phenotype was associated with a homozygous missense mutation in exon 5.


Osteogenic differentiation of mouse embryonic stem cells: differential gene expression analysis by cDNA microarray and purification of osteoblasts by cadherin-11 magnetically activated cell sorting.
We have previously shown osteogenic differentiation of mouse embryonic stem (ES) cells and temporal enrichment with osteoblastic cells, by stimulation with serum-containing culture medium supplemented with beta-glycerophosphate, ascorbate, and dexamethasone. In our present study we have used similar culture conditions to further investigate osteogenic differentiation of mouse ES cells. Using reverse transcription-polymerase chain reaction (RT-PCR) we demonstrated the expression of genes associated with osteoblast differentiation including the bone matrix protein osteocalcin and the transcription factor Cbfa-1/runx2. Furthermore, results of cDNA microarray analysis, and subsequent RT-PCR analysis of differentiating ES cells after exposure to osteogenic stimuli, revealed a combination of upregulation of genes involved in osteoblast differentiation including osteopontin, HSP-47, and IGF-II coupled with downregulation of genes involved in differentiation of other phenotypes such as the neuroectoderm factor Stra-13. Finally, we have applied magnetically activated cell-sorting methods to ES cell cultures treated with osteogenic stimuli and, using an antibody to cadherin-11, have purified a subpopulation of cells with osteoblastic characteristics.


Atom number in magneto-optic traps with millimeter scale laser beams.
We measure the number of atoms N trapped in a conventional vapor-cell magneto-optic trap (MOT) using beams that have a diameter d in the range 1-5 mm. We show that the N is proportional to d(3.6) scaling law observed for larger MOTs is a robust approximation for optimized MOTs with beam diameters as small as 3 mm. For smaller beams, the description of the scaling depends on how d is defined. The most consistent picture of the scaling is obtained when d is defined as the diameter where the intensity profile of the trapping beams decreases to the saturation intensity. Using this definition, N scales as d(6) for d<2.3 mm but, at larger d, N still scales as d(3.6).


Alpha-gamma hybrid peptides that contain the conformationally constrained gabapentin residue: characterization of mimetics of chain reversals.
The crystal structures of four dipeptides that contain the stereochemically constrained gamma-amino acid residue gabapentin (1-(aminomethyl)cyclohexaneacetic acid Gpn) are described. The molecular conformation of Piv-Pro-Gpn-OH (1), reveals a beta-turn mimetic conformation, stabilized by a ten atom C[bond]H...O hydrogen bond between the Piv CO group and the pro S hydrogen of the Gpn CH(2)[bond]CO group. The peptides Boc-Gly-Gpn-OH (2), Boc-Aib-Gpn-OH (3), and Boc-Aib-Gpn-OMe (4) form compact, folded structures, in which a distinct reversal of polypeptide chain direction is observed. In all cases, the Gpn residue adopts a gauche,gauche (g,g) conformation about the C(gamma)[bond]C(beta) (theta(1)) and C(beta)[bond]C(alpha) (theta(2)) bonds. Two distinct Gpn conformational families are observed. In peptides 1 and 3, the average backbone torsion angle values for the Gpn residue are phi=98 degrees, theta(1)=-62 degrees, theta(2)=-73 degrees, and psi=79 degrees, while in peptide 2 and 4 the average values are phi=-103 degrees, theta(1)=-46 degrees, theta(2)=-49 degrees, and psi=-92 degrees. In the case of 1 and 3, an intramolecular nine-membered O[bond]H...O hydrogen bond is formed between the C[double bond]O of the preceding residue and the terminal carboxylic acid OH group. All four alpha-gamma dipeptide sequences yield compact folded backbone conformations; this suggests that the Gpn residue may be employed successfully in the design of novel folded structures.


Growth hormone, skeletal growth factors and osteoporosis.
Bone remodeling is regulated by systemic hormones and growth factors acting in concert to maintain normal bone mass. Growth hormone has important effects on bone metabolism and acts by stimulating the production of insulin-like growth factor (IGF) I in the liver. Conditions of growth hormone deficiency have decreased bone mass that could be modified by growth hormone administration. Skeletal cells synthesize a variety of growth factors with diverse properties and levels of regulation. IGF I and II seem to play a major role in the local regulation of bone remodeling, and other growth factors have important mitogenic properties. Skeletal growth factors may be regulated at the level of synthesis, activation, receptor binding or binding proteins. Growth factors such as IGF I could be considered for the systemic treatment of various bone disorders, or drugs could be developed to modify the synthesis or activity of the locally produced factors. Additional research should result in the development of novel approaches to the treatment of osteoporosis.


[Maintenance of the cranial cruciate ligament rupture in the dog using an intra-articular ("over-the-top") and an extra-articular ("fascia-doubling") method].
The present study compares the results of an intraarticular ("over-the-top") and an extraarticular (fascia lata imbrication) surgical method to treat the rupture of the cranial cruciate ligament in dogs. In a period of 18 months, 132 stifle joints of 125 dogs with a partial or complete rupture of the cranial cruciate ligament were examined according to a systematic protocol at the Clinic for Small Animals of the Hannover School of Veterinary Medicine, Hannover, Germany. Surgery was performed with either one or the other of the two surgical techniques. In all cases a " cleaning up " was carried out. If the medial meniscus was damaged, it was resected and the lesion was investigated. The imbrication technique of the lateral aspect of the fascia lata was easier and faster to perform. Using the "over-the-top" method, there were difficulties to guide the fascial strip through the intercondylar notch. After six months 88 patients were re-examined clinically and radiologically according to the same protocol as mentioned above. In the present study it was demonstrated that patients treated with the extraarticular method showed a higher rate (76.5%) of normal limb function after six months and walked earlier without lameness compared with dogs operated with the "over-the-top" technique (41.5%). The same results were valid for patients of giant breeds. Patients operated with the imbrication technique showed more arthrosis of the patellar tip. After six months stability, evaluated with the cranial drawer test and the tibia compression test was satisfying with both techniques.


Gene expression and integrated stress response in HepG2/C3A cells cultured in amino acid deficient medium.
The integrated stress response (ISR), a defense mechanism cells employ when under stress (e.g., amino acid deprivation), causes suppression of global protein synthesis along with the paradoxical increased expression of a host of proteins that are useful in combating various stresses. Genes that were similarly differentially expressed under conditions of either leucine- or cysteine-depletion were identified. Many of the genes known to contain an amino acid response element and to be induced in response to eIF2α phosphorylation and ATF4 heterodimer binding (ATF3, C/EBPβ, SLC7A1, SLC7A11, and TRIB3), as well as others shown to be induced downstream of eIF2α phosphorylation (C/EBPγ, CARS, SARS, CLCN3, CBX4, and PPP1R15A) were among the upregulated genes. Evidence for the induction of the ISR in these cells also included the increased phosphorylation of eIF2α and increased protein abundance of ATF4, ATF3, and ASNS in cysteine- and leucine-depleted cells. Based on genes highly differentially expressed in both leucine- and cysteine-deficient cells, a list of 67 downregulated and 53 upregulated genes is suggested as likely targets of essential amino acid deprivation in mammalian cells.


Epigenetic markers for newborn congenital heart defect (CHD).
Our objective was to determine whether there were significant differences in genome-wide DNA methylation in newborns with major congenital heart defect (CHD) compared to controls. We also evaluated methylation of cytosines in CpG motifs for the detection of these CHDs. Genome-wide DNA methylation analysis was performed on DNA from 60 newborns with various CHDs, including hypoplastic left heart syndrome, ventricular septal deficit, atrial septal defect, pulmonary stenosis, coarctation of the aorta and Tetralogy of Fallot, and 32 controls. Highly significant differences in cytosine methylation were seen in a large number of genes throughout the genome for all CHD categories. Gene ontology analysis of CHD overall indicated over-represented biological processes involving cell development and differentiation, and anatomical structure morphogenesis. Methylation of individual cytosines in CpG motifs had high diagnostic accuracy for the detection of CHD. For example, for coarctation one predictive model based on levels of particular cytosine nucleotides achieved a sensitivity of 100% and specificity of 93.8% (AUC = 0.974, p < 0.00001). Profound differences in cytosine methylation were observed in hundreds of genes in newborns with different types of CHD. There appears to be the potential for development of accurate genetic biomarkers for CHD detection in newborns.


Relation between bone mineral density and IL-17 serum levels in Serbian patients with early Rheumatoid arthritis.
Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial inflammation and destruction of joint cartilage and bone. Different cytokines play important role in the processes that cause articular destruction and extra-articular manifestations in RA. The contribution of cytokines representing the Th1 (INF-γ), Th2 (IL-4) and IL-17A to the pathogenesis of early RA and bone mineral density (BMD) loss in still poorly understood. Serum samples of 38 early RA patients were evaluated for erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), C-reactive protein (CRP), anti-cyclic citrullinated peptide antibodies (anti-CCP) and for the tested cytokines (IL-17A, IL-4 and INF-γ). BMD was evaluated by dualenergy X-ray absorptiometry (DXA). Disease activity score (DAS28) calculation was assessed for all patients. Control serum samples were obtained from 34 healthy volunteers. The levels of tested cytokines were significantly higher (IL-17A, p<0.001; INF-γ, P<0.001; IL-4, P<0.01) in patients with early RA, compared to the healthy controls. In early RA patients, strong correlation of serum IL-17A was found with DAS28, ESR and CRP. Also, a significant negative correlation was found between serum INF-γ levels and the DAS28 score. Significantly positive correlation of BMD values and CRP, DAS28 IL-17A were also demonstrated. DXA analysis revealed that the most common site for osteoporosis was the lumbar spine followed by the femoral neck. BMD values significantly correlated with CRP, DAS28 score and IL-17A serum levels. The mean serum IL-17A levels, in patients with early RA, corresponded with disease activity, severity and BMD loss, indicating the potential usefulness of serum IL-17A in defining the disease activity and bone remodeling.


[Initial Clinical Experiences Using Ocriplasmin for the Treatment of Vitreomacular Traction with or without a Macular Hole].
In vitreomacular traction (VMT), there is abnormal adhesion between the vitreous cortex and the retina, especially in the fovea. Symptoms of VMT include metamorphopsia and a decrease in visual acuity. Since 2013, ocriplasmin (Jetrea®) has been approved for treatment of symptomatic vitreomacular traction with or without macular holes (≤ 400 µm). We retrospectively examined twenty-three eyes of twenty-one patients who underwent intravitreal ocriplasmin treatment for symptomatic vitreomacular traction with or without macular holes. Best corrected visual acuity and central retinal thickness (CRT) were measured in advance and after ocriplasmin treatment. The numbers of resolved vitreomacular traction and closed macular holes were documented. Vitreomacular traction was resolved in eight of twenty-three eyes (34.8 %); in fifteen eyes (65.2 %) it was persistent and two of four macular holes were found closed. The average best corrected visual acuity was 0.39 ± 0.25 logMAR at baseline and 0.41 ± 0.24 logMAR at the first follow-up visit after injection (p = 0.613). The average CRT was 453.3 ± 172.7 µm at baseline, with a slight decrease to 412.0 ± 212 µm (p = 0.124). Intravitreal injection of ocriplasmin appears is an experimental therapy in patients with symptomatic vitreomacular traction. Patient selection seems to be critically important for the therapeutic outcome, whereas greater age, specific VMT morphology and missing chromatopsia seem to be negative predictors.


Do reciprocal interactions between cell stress proteins and cytokines create a new intra-/extra-cellular signalling nexus?
Cytokine biology began in the 1950s, and by 1988, a large number of cytokines, with a myriad of biological actions, had been discovered. In 1988, the basis of the protein chaperoning function of the heat shock, or cell stress, proteins was identified, and it was assumed that this was their major activity. However, since this time, evidence has accumulated to show that cell stress proteins are secreted by cells and can stimulate cellular cytokine synthesis with the generation of pro- and/or anti-inflammatory cytokine networks. Cell stress can also control cytokine synthesis, and cytokines are able to induce, or even inhibit, the synthesis of selected cell stress proteins and may also promote their release. How cell stress proteins control the formation of cytokines is not understood and how cytokines control cell stress protein synthesis depends on the cellular compartment experiencing stress, with cytoplasmic heat shock factor 1 (HSF1) having a variety of actions on cytokine gene transcription. The endoplasmic reticulum unfolded protein response also exhibits a complex set of behaviours in terms of control of cytokine synthesis. In addition, individual intracellular cell stress proteins, such as Hsp27 and Hsp90, have major roles in controlling cellular responses to cytokines and in controlling cytokine synthesis in response to exogenous factors. While still confusing, the literature supports the hypothesis that cell stress proteins and cytokines may generate complex intra- and extra-cellular networks, which function in the control of cells to external and internal stressors and suggests the cell stress response as a key parameter in cytokine network generation and, as a consequence, in control of immunity.


Elevated catalase expression in a fungal pathogen is a double-edged sword of iron.
Most fungal pathogens of humans display robust protective oxidative stress responses that contribute to their pathogenicity. The induction of enzymes that detoxify reactive oxygen species (ROS) is an essential component of these responses. We showed previously that ectopic expression of the heme-containing catalase enzyme in Candida albicans enhances resistance to oxidative stress, combinatorial oxidative plus cationic stress, and phagocytic killing. Clearly ectopic catalase expression confers fitness advantages in the presence of stress, and therefore in this study we tested whether it enhances fitness in the absence of stress. We addressed this using a set of congenic barcoded C. albicans strains that include doxycycline-conditional tetON-CAT1 expressors. We show that high basal catalase levels, rather than CAT1 induction following stress imposition, reduce ROS accumulation and cell death, thereby promoting resistance to acute peroxide or combinatorial stress. This conclusion is reinforced by our analyses of phenotypically diverse clinical isolates and the impact of stochastic variation in catalase expression upon stress resistance in genetically homogeneous C. albicans populations. Accordingly, cat1Δ cells are more sensitive to neutrophil killing. However, we find that catalase inactivation does not attenuate C. albicans virulence in mouse or invertebrate models of systemic candidiasis. Furthermore, our direct comparisons of fitness in vitro using isogenic barcoded CAT1, cat1Δ and tetON-CAT1 strains show that, while ectopic catalase expression confers a fitness advantage during peroxide stress, it confers a fitness defect in the absence of stress. This fitness defect is suppressed by iron supplementation. Also high basal catalase levels induce key iron assimilatory functions (CFL5, FET3, FRP1, FTR1). We conclude that while high basal catalase levels enhance peroxide stress resistance, they place pressure on iron homeostasis through an elevated cellular demand for iron, thereby reducing the fitness of C. albicans in iron-limiting tissues within the host.


Communication and computation in the central nervous system.
Some aspects of the communicational and computational features of the central nervous system (CNS) are presented. We have suggested the existence in the CNS of two types of electrochemical transmission, namely the wiring transmission (WT) and the volume transmission (VT), where WT is the classical type of neural transmission based on the physical substrate of neural chains, whereas VT consists of chemical signals diffusing in the extracellular fluid and reaching receptors on neuronal cells located far away from the signal release site. Experimental data supporting this hypothesis as well as the functional characteristics of both types of transmission are reported.


Comparison of microcoils and polyvinyl alcohol particles in selective microcatheter angioembolization of non variceal acute gastrointestinal hemorrhage.
To compare the efficacy of polyvinyl alcohol (PVA) particles with microcoils in angiembolisation of non variceal acute gastrointestinal haemorrhage. This is a retrospective cross-sectional study of patients who underwent transcatheter angioembolization from January, 1995 to December, 2013 at Aga Khan University Hospital, Karachi. Patients were divided into two groups on basis of use of either microcoils or PVA particles and compared in terms of technical success, clinical success, re-bleeding and ischemic complication rates. Chi (χ(2)) square and Fisher's exact tests were applied and a P-value of less than 0.05 was considered statistically significant. Fifty seven patients underwent angioembolization. Microcoil and PVA particles embolization was performed in 63% (36/57) and 35% (20/57) cases respectively. Technical success was achieved in all cases (100%). Clinical success rate was higher in microcoils group (92%) than PVA particles group (75%) with statistically significant P value (p=0.048). Ischemic complication was seen in one case (3%) in the microcoil group, while no such complications were seen in the PVA particles group. In angioembolization of non variceal acute gastrointestinal haemorrhage microcoils are better than Polyvinyl alcohol particles with higher clinical success and lower re-bleed rates.


Human-mediated emergence as a weed and invasive radiation in the wild of the CD genome allotetraploid rice species (Oryza, Poaceae) in the Neotropics.
The genus Oryza is being used as a model in plant genomic studies although there are several issues still to be resolved regarding the spatio-temporal evolution of this ancient genus. Particularly contentious is whether undated transoceanic natural dispersal or recent human interference has been the principal agent determining its present distribution and differentiation. In this context, we studied the origin and distribution history of the allotetraploid CD rice genome. It is endemic to the Neotropics but the genus is thought to have originated in the Paleotropics, and there is relatively little genetic divergence between some orthologous sequences of the C genome component and their Old World counterparts. Because of its allotetraploidy, there are several potential pitfalls in trying to date the formation of the CD genome using molecular data and this could lead to erroneous estimates. Therefore, we rather chose to rely on historical evidence to determine whether or not the CD genome was present in the Neotropics before the arrival of Columbus. We searched early collections of herbarium specimens and studied the reports of explorers of the tropical Americas for references to rice. In spite of numerous collectors traveling inland and collecting Oryza, plants determined as CD genome species were not observed away from cultivated rice fields until 1869. Various arguments suggest that they only consisted of weedy forms until that time. The spatio-temporal distribution of herbarium collections fits a simple biogeographical scenario for the emergence in cultivated rice fields followed by radiation in the wild of the CD genome in the Neotropics during the last four centuries. This probably occurred from species introduced to the Americas by humans and we found no evidence that the CD genome pre-existed in the Old World. We therefore propose a new evolutionary hypothesis for such a recent origin of the CD genome. Moreover, we exemplify how an historical approach can provide potentially important information and help to disentangle the timing of evolutionary events in the history of the Oryza genomes.


Modeling of hollow-fiber capillary reactor for the production of L-Alanine with coenzyme regeneration.
Continuous production of L-alanine with conjugated enzyme systems of alanine dehydrogenase (AlaDH) and lactate dehydrogenase (LDH) or alcohol dehydrogenase (ADH) was carried out with NAD regeneration in an ultrafiltration hollow-fiber capillary reactor (HFCR) which was proposed as a test bioreactor with very small scale. In the AlaDH/LDH system, pyruvate is the intermediate product for L-alanine so that an optimal point existed in pyruvate concentration for the production rate of L-alanine. NAD cycling number of 4850 and L-alanine productivity of 61.7 mmol/L h were obtained at the best condition. In the AlaDH/ADH system, however, the substrate inhibition in the AlaDH reaction by pyruvate should be considered and the best results of NAD cycling number and (L)-alanine productivity were 2700 and 13.5 mmol/L h, respectively. In consideration of concentration distribution and mixing in the axial direction on an HFCR, performance of the reactor was theoretically analyzed with a multistage stirred tank reactor model combined with the kinetic model based on all the elementary reactions involved. Although quantitative discrepancy existed in some cases, the present theoretical model could explain experimental results and is expected to be generally applicable to standard hollow fiber reactors.


Molecular mechanisms of cell-cell interaction in Dictyostelium discoideum.
During development of the cellular slime mold Dictyostelium discoideum, cells migrate in response to cAMP to form aggregates, which give rise to fruiting bodies consisting of two major cell types: spores and stalk cells. Multicellularity is achieved by the expression of two types of cell-cell adhesion sites. The EDTA-sensitive binding sites are expressed at the initial stage of development. At the aggregation stage, cells acquire EDTA-resistant binding sites, which are mediated by a cell-surface glycoprotein of Mr80,000 (gp80). gp80 is preferentially associated with cell surface filopodia, which are probably involved in the initiation of contact formation between cells. Covaspheres conjugated with gp80 bind specifically to aggregation-stage cells. The binding can be inhibited by precoating cells with an anti-gp80 monoclonal antibody, thus suggesting that gp80 mediates cell-cell binding via homophilic interaction. The structure of gp80 predicted from its cDNA sequence can be divided into three major domains: a membrane anchor, a hinge, and a globular region. An analysis of fusion proteins containing different gp80 segments shows that the cell-binding activity resides in the globular region. In the postaggregation stages, gp80 is replaced by other surface glycoproteins in maintaining cell-cell adhesion. One of them has a Mr of 150,000 (gp150). Anti-gp150 antibodies have no effect on aggregation-stage cells, but they disrupt cell-cell adhesion at subsequent stages. It becomes evident that the complex phenomena of cell adhesion and tissue organization involve the participation of a number of surface glycoproteins.


Accurate Thermochemistry for Main-Group Elements up to Xenon with the Wn-P34 Series of Composite Methods.
In the present study, we introduce the accurate Wn-P34 quantum chemistry composite methods with applicability to heavy p-block elements up to xenon. For a set of thermochemical properties for prototypical third- and fourth-row species and for a diverse set of small light-main-group species, they show accuracies of ∼3 kJ mol-1 or better. Overall, the Wn-P34 methods are comparable in accuracy to Wn, with a widened applicability to heavier elements. We have used Wn-P34 to compile the P34 set of accurate thermochemical values for heavy p-block species, and we have applied this set to assess a wide range of lower-cost methods. The results of our assessment show that the G4(MP2)-XK composite method provides adequate treatments for these species, but several widely used double-hybrid density functional theory (DH-DFT) methods show uncharacteristically large deviations. In contrast, we find it presently surprising that some pure and hybrid DFT methods such as TPSS and SCANh perform quite well. We hope that our findings and new tools would facilitate the application of computational chemistry for heavy elements, of which the properties are yet to be broadly explored.


EUS-guided Gastrojejunostomy Versus Laparoscopic Gastrojejunostomy: An International Collaborative Study.
Endoscopic ultrasound-guided gastrojejunostomy (EUS-GJ) with placement of a lumen-apposing metal stent is a minimally invasive and efficacious procedure for gastric outlet obstruction (GOO) patients not amenable to surgery. Laparoscopic gastrojejunostomy (Lap-GJ) has traditionally been the standard of care. No direct comparison between EUS-GJ and Lap-GJ has been described. Our aim was to compare the clinical outcomes, success rate, and adverse events (AE) of EUS-GJ with Lap-GJ. Patients with GOO from 4 academic centers in 3 countries were included. Technical success was defined as successful creation of a gastrojejunostomy. Clinical success was defined as the ability to tolerate a diet postprocedure. Data were collected on 54 patients. A total of 25 patients underwent EUS-GJ (male n=11, mean age 63.9 y) and 29 patients underwent Lap-GJ (male n=22, mean age 75.8 y). Technical success was achieved in 29 (100%) Lap-GJ group patients and 23 (88%) in the EUS-GJ group (P=0.11). AEs occurred in 41% (n=12) of patients in the Lap-GJ group and 12% (n=3) in the EUS-GJ group (P=0.0386). According to the Clavien-Dindo Classification, the Lap-GJ group AEs were grade I (n=4), grade II (n=5), grade III (n=2), and grade V (n=1); the EUS-GJ AEs were grade II (n=2) and grade V (n=1). Although the EUS-GJ group contained more complex patients, efficacy was similar between the groups. AEs were significantly lower in the EUS-GJ group. EUS-GJ is a safe and efficacious, minimally invasive option for patients with GOO.


Using anisotropic and narrowband ambient noise for continuous measurements of relative clock drift between independent acoustic recorders.
Relative clock drift between instruments can be an issue for coherent processing of acoustic signals, which requires data to be time-synchronized between channels. This work shows how cross correlation of anisotropic narrowband ambient noise allows continuous estimation of the relative clock drift between independent acoustic recorders, under the assumption that the spatial distribution of the coherent noise sources is stationary. This method is applied to two pairs of commercial passive acoustic recorders deployed up to 14 m apart at 6 and 12 m depth, respectively, over a period of 10 days. Occasional calibration signals show that this method allows time-synchronizing the instruments to within ±1 ms. In addition to a large linear clock drift component on the order of tens of milliseconds per hour, the results reveal for these instruments non-linear excursions of up to 50 ms that cannot be measured by standard methods but are crucial for coherent processing. The noise field displays the highest coherence between 50 and 100 Hz, a bandwidth dominated by what are believed to be croaker fish, which are particularly vocal in the evenings. Both the passive and continuous nature of this method provide advantages over time-synchronization using active sources.


Syntheses and structures of two new coordination polymers generated from a 4-aminotriazole-bridged organic ligand and CoII salts.
Organic ligands and counter-anions influence the coordination spheres of metal cations and hence the construction of coordination polymers (CPs). The specific bent geometries of five-membered heterocyclic triazole bridging organic ligands are capable of generating CPs with novel patterns not easily obtained using rigid linear ligands. A multidentate 4-aminotriazole-bridged organic ligand, namely 4-amino-3,5-bis(4,3'-bipyridyl-5'-yl)-4H-1,2,4-triazole (L) has been prepared and used to synthesize two CoII coordination polymers, namely poly[[[μ2-4-amino-3,5-bis(4,3'-bipyridyl-5'-yl)-4H-1,2,4-triazole-κ2N:N']bis(methanol-κO)cobalt(II)] bis(perchlorate)], {[Co(C22H16N8)2(CH3OH)2](ClO4)2}n, (I), and poly[[μ3-4-amino-3,5-bis(4,3'-bipyridyl-5'-yl)-4H-1,2,4-triazole-κ3N:N':N'']dichloridocobalt(II)], [CoCl2(C22H16N8)]n, (II), using CoX2 salts [X = ClO4 for (I) and Cl for (II)] under solvothermal conditions. Single-crystal X-ray structure analysis revealed that they both feature two-dimensional networks. Cobalt is located on an inversion centre in (I) and in a general position in (II). In (I), L functions as a bidentate cis-conformation ligand linking CoII ions, while it functions as a tridentate trans-conformation linker binding CoII ions in (II). In addition, O-H...N and N-H...O hydrogen bonds and C-H...π interactions exist in (I), while N-H...Cl and π-π interactions exist in (II), and these weak interactions play an important role in aligning the two-dimensional nets of (I) and (II) in the solid state. As the compounds were synthesized under the same conditions, the significant structural variations between (I) and (II) are believed to be determined by the different sizes and coordination abilities of the counter-anions. IR spectroscopy and diffuse reflectance UV-Vis spectra were also used to investigate the title compounds.


[Prevention of acute respiratory platinosis].
289 respiratory platinosis cases were studied to assess the effectiveness of the preventive treatment differentiated depending on the form (immune/non-immune) and gravity (mild, moderate and severe). The treatment was performed during excessive exposition to the salts of the platinum group metals, and proved to be highly effective (76.9% in acute and 100% in light cases) for the prevention of clinical manifestations of the disease.


[Clinical observations on the rheological and hemodynamic effects of some drugs acting on the microcirculation].
A clinical research was carried out on the rheological and haemodynamic effects of various drugs commonly employed in the treatment of certain disturbances of the microcirculation. In addition to low molecular weight dextrane, a comparison was also made of three other substances theoretically capable of interfering with the fundamental relationship between viscosity, volume and haemodynamics. Assessment of the clinical signs was made both via evaluation of the peripheral haemodynamics during rest and during post-ischaemic hyperaemia and through determination of changes in blood viscosity by means of an original capillary viscosimeter. Some conclusions of a certain speculative and practical interest are drawn from the research as a whole.


Antiviral Treatment Guidelines for Middle East Respiratory Syndrome.
Middle East respiratory syndrome (MERS) is an acute infectious disease of the respiratory system caused by the new betacoronavirus (MERS coronavirus, MERS-CoV), which shows high mortality rates. The typical symptoms of MERS are fever, cough, and shortness of breath, and it is often accompanied by pneumonia. The MERS-CoV was introduced to Republic of Korea in May 2015 by a patient returning from Saudi Arabia. The disease spread mostly through hospital infections, and by the time the epidemic ended in August, the total number of confirmed diagnoses was 186, among which 36 patients died. Reflecting the latest evidence for antiviral drugs in the treatment of MERS-CoV infection and the experiences of treating MERS patients in Republic of Korea, these guidelines focus on antiviral drugs to achieve effective treatment of MERS-CoV infections.


Metabolic risk factors and arterial stiffness in Indian children of parents with metabolic syndrome.
To investigate the possible association between metabolic syndrome (MS) and arterial stiffness in Indian children with parental MS status. A cross-sectional study was conducted in 140 overweight/obese and 60 normal-weight Indian children (mean age, 11.4 ± 2.8 years) along with one of their parents during 2008-2009. Data on weight, height, blood pressure, serum lipids, zinc, insulin, and glucose were collected. Intima media thickness (CIMT) and stiffness parameters were assessed in the right carotid artery. Physical activity and diet were assessed using structured questionnaires. Body composition was measured using dual-energy x-ray absorptiometry. A gradual increase in the percentage of MS children with an increasing number of MS components in parents was observed. Mean values for arterial stiffness, pulse wave velocity, and elastic modulus were significantly higher in MS children of MS parents than in MS children of normal parents (p < 0.05). A significant correlation was observed for lifestyle, metabolic, and arterial parameters among child-parent pairs (p < 0.05). Multiple logistic regression revealed that children's CIMT and arterial stiffness were significantly associated (p < 0.01) with their serum levels of triglycerides, high-density lipoprotein, and zinc, as well as with parental MS-CIMT. Parental MS status and lifestyle factors increase the risk of MS and arterial abnormalities in children.


System-Wide Action Team (SWAT)--a new level of collaboration.
A multidisciplinary System-Wide Action Team was coordinated by nursing leadership in response to increasing patient census and acuity. The primary objectives of the team were to develop mechanisms that would expedite patient's admissions and discharges, communicate key messages to all concerned staff, support each other, and communicate announcements about topics or urgent issues that relate to managing the hospital. The team has implemented improvement initiatives around diagnostic testing and scheduling, expediting the admission process, reducing discharge delays, and staffing to hospital census demands.


Cloning, expression, and analysis of the group 2 allergen from Dermatophagoides farinae from China.
To obtain the recombinant group 2 allergen product of Dermatophagoides farinae (Der f 2), the Der f 2 gene was synthesized by RT-PCR. The full-length cDNA comprised 441 nucleotides and was 99.3% identical to the reference sequence (GenBank AB195580). The cDNA was bound to vector pET28a to construct plasmid pET28a(+)-Der f 2, which was transformed into E. coli BL21 and induced by IPTG. SDS-PAGE showed a specific band of about 14kDa in the hole cell lysate. As estimated by chromatography, about 3.86 g of the recombinant product as obtained, which conjugated with serum IgE from asthmatic children. The protein had a signal peptide of 17 amino acids. Its secondary structure comprised an alpha helix (19.86%), an extended strand (30.82%), and a random coil (49.32%). The subcellular localization of this allergen was predicted to be at mitochondria. Furthermore, its function was shown to be associated with an MD-2-related lipid-recognition (ML) domain. The results of this study provide a solid foundation for large-scale production of the allergen for clinical diagnosis and treatment of allergic disorders.


High nuc DNA load in whole blood is associated with sepsis, mortality and immune dysregulation in Staphylococcus aureus bacteraemia.
Staphylococcus aureus bacteraemia is a disease with varying presentation, ranging from uncomplicated to life-threatening infections. In S. aureus bacteraemia, a high load of bacterial DNA in blood has been linked to mortality. We hypothesized that a high DNA load would also be linked to the presence of sepsis, and to high C-reactive protein (CRP) and lymphopaenia, indicating inflammation and immunosuppression. Twenty-seven patients with culture-proven S. aureus bacteraemia, 13 (48%) with sepsis and six (22%) non-survivors, were enrolled in a prospective study. Blood samples were collected on days 0, 1-2, 3-4, 6-8, 13-15 and 26-30, and subjected to droplet digital PCR targeting the nuc gene to determine the nuc DNA load. nuc DNA was detected on days 0-2 in 22 patients (81%), and on days 6-8 in three patients (all non-survivors). The nuc DNA load on days 1-2 was significantly elevated in patients with sepsis (median 2.69 versus 1.32 log10 copies/mL; p = .014) and in non-survivors (median 2.5 versus 1.0 log10 copies/mL; p = .033). Patients with a high nuc DNA load (>3.0 log10 copies/mL) on days 1-2 had significantly elevated CRP levels at all timepoints, and significantly decreased lymphocyte counts on days 0, 1-2, 13-15 and 26-30. Our results indicate that a high initial load of S. aureus DNA in blood is associated with sepsis, mortality and persistent immune dysregulation in S. aureus bacteraemia patients. Further studies are needed to define the role of bacterial DNA load monitoring in the management of S. aureus bacteraemia.


Genetic variant in 8q24 is associated with prognosis for gastric cancer in a Chinese population.
Single nucleotide polymorphisms (SNPs) located in long noncoding RNA CASC8 gene may influence the process of splicing and stability of messenger RNA conformation, resulting in the modification of its interacting partners. Genome-wide association studies have identified the SNP rs10505477 and SNP rs1562430 in CASC8 were associated with risk of the colorectal cancer (CRC) and breast cancer, respectively. In the present study, we genotyped the 940 surgically resected gastric cancer patients to explore the association between these two SNPs (e.g., rs10505477 and rs1562430) and survival of gastric cancer in a Chinese population. We found that the patients carrying rs10505477 GG genotype survived for a longer time than those with the GA and AA genotypes (log-rank P = 0.030). The similar result was also found in the dominant model (GA/AA vs GG, HR = 1.32, 95% CI = 1.08-1.63, log-rank P = 0.008). This risk effect was more pronounced among patients with tumor size ≤ 5 cm, diffuse-type gastric cancer, lymph node metastasis, no distant metastasis, and TNM stage III and IV. Furthermore, the area under the curve at five years was dramatically increased from 0.619 to 0.624 after adding the rs10505477 risk score to the traditional clinical risk score (TNM stage and lymph node metastasis). However, there was no association be found between the rs1562430 and the survival of gastric cancer. These findings suggested the SNP rs10505477 may contribute to the survival of gastric cancer and be a potential prognostic biomarker of gastric cancer.


The remarkable hydration of the antifreeze protein Maxi: a computational study.
The long four-helix bundle antifreeze protein Maxi contains an unusual core for a globular protein. More than 400 ordered waters between the helices form a nano-pore of internal water about 150 Å long. Molecular dynamics simulations of hydrated Maxi were carried out using the CHARMM27 protein forcefield and the TIP3P water model. Solvation in the core and non-core first hydration shell was analyzed in terms of water-water H-bond distance-angle distributions. The core had an increased population of low-angle H-bonds between water pairs relative to bulk water. Enhancement of low angle H-bonds was particularly pronounced for water pairs at the interfaces between apolar and polar regions inside the protein core, characteristic of the anchored clathrate solvation structure seen previously in the ice-nuclei binding surfaces of type I, type III, and beta-helical antifreeze proteins. Anchored clathrate type solvation structure was not seen in the exterior solvation shell except around residues implicated in ice binding. Analysis of solvation dynamics using water residence times and diffusion constants showed that exterior solvation shell waters exchanged rapidly with bulk water, with no difference between ice-binding and non-binding residues. Core waters had about ten-fold slower diffusion than bulk water. Water residence times around core residues averaged about 8 ps, similar to those on the exterior surface, but they tended to exchange primarily with other core water, resulting in longer, >40 ps residence times within the core. Preferential exchange or diffusion of the water along the long axis of the water core of Maxi was not detected.


Synthesis and Properties of 6-Aryl-4-azidocinnolines and 6-Aryl-4-(1,2,3-1H-triazol-1-yl)cinnolines.
An efficient approach towards the synthesis of 6-aryl-4-azidocinnolines was developed with the aim of exploring the photophysical properties of 6-aryl-4-azidocinnolines and their click reaction products with alkynes, 6-aryl-4-(1,2,3-1H-triazol-1-yl)cinnolines. The synthetic route is based on the Richter-type cyclization of 2-ethynyl-4-aryltriazenes with the formation of 4-bromo-6-arylcinnolines and nucleophilic substitution of a bromine atom with an azide functional group. The developed synthetic approach is tolerant to variations of functional groups on the aryl moiety. The resulting azidocinnolines were found to be reactive in both CuAAC with terminal alkynes and SPAAC with diazacyclononyne, yielding 4-triazolylcinnolines. It was found that 4-azido-6-arylcinnolines possess weak fluorescent properties, while conversion of the azido function into a triazole ring led to complete fluorescence quenching. The lack of fluorescence in triazoles could be explained by the non-planar structure of triazolylcinnolines and a possible photoinduced electron transfer (PET) mechanism. Among the series of 4-triazolylcinnoline derivatives a compound bearing hydroxyalkyl substituent at triazole ring was found to be cytotoxic to HeLa cells.


Bone Marrow Assessment in Liver Cirrhosis Patients with Otherwise Unexplained Peripheral Blood Cytopenia.
We performed a retrospective single-center analysis to investigate the diagnostic yield of bone marrow puncture in patients with liver cirrhosis and cytopenia. Liver cirrhosis patients receiving bone marrow aspiration or biopsy for the diagnostic work-up of otherwise unexplained peripheral blood cytopenia at our institution between 2004 and 2020 were enrolled in this study. We evaluated findings from cytologic, histologic and immunologic assessment and final diagnostic outcomes. A total of 118 patients with a median age of 55 years and a median Child-Pugh score of B (8 points) were enrolled. The main etiologies of liver cirrhosis were viral hepatitis (B and C) or chronic alcohol consumption. The majority of patients (60%) exhibited concurrent anemia, leukocytopenia and thrombocytopenia. Bone marrow assessment revealed normal, unspecific or reactive alterations in 117 out of 118 patients (99%). One patient was diagnosed with myelodysplastic syndrome. Our findings suggest that peripheral blood cytopenia in patients with liver cirrhosis is rarely associated with a primary bone marrow pathology.


Excessive oozing through the fabric of the branched Cryolife-Jotec Evita Open NEO hybrid prosthesis.
We report 3 cases of excessive oozing after aortic arch replacement using the frozen elephant trunk technique through the fabric of the branched Cryolife-Jotec Evita Open NEO hybrid prosthesis. After weaning from cardiopulmonary bypass, protamine administration as well as standard substitution of plasmatic and cellular coagulation adjusted to thrombelastometry, excessive oozing persisted leading to a complicated postoperative course in all 3 patients.


Charging dynamics of an individual nanopore.
Meso-porous electrodes (pore width « 1 µm) are a central component in electrochemical energy storage devices and related technologies, based on the capacitive nature of electric double-layers at their surfaces. This requires that such charging, limited by ion transport within the pores, is attained over the device operation time. Here we measure directly electric double layer charging within individual nano-slits, formed between gold and mica surfaces in a surface force balance, by monitoring transient surface forces in response to an applied electric potential. We find that the nano-slit charging time is of order 1 s (far slower than the time of order 3 × 10-2 s characteristic of charging an unconfined surface in our configuration), increasing at smaller slit thickness, and decreasing with solution ion concentration. The results enable us to examine critically the nanopore charging dynamics, and indicate how to probe such charging in different conditions and aqueous environments.


Current Synthesis and Systematic Review of Main Effects of Calf Blood Deproteinized Medicine (Actovegin®) in Ischemic Stroke.
Stroke is one of the largest problems and clinical-social challenges within neurology and, in general, pathology. Here, we briefly reviewed the main pathophysiological mechanisms of ischemic stroke, which represent targets for medical interventions, including for a calf blood deproteinized hemodialysate/ultrafiltrate. We conducted a systematic review of current related literature concerning the effects of Actovegin®, of mainly the pleiotropic type, applied to the injury pathways of ischemic stroke. The bibliographic resources regarding the use of Actovegin® in ischemic stroke are scarce. The main Actovegin® actions refer to the ischemic stroke lesion items' ensemble, targeting tissue oxidation, energy metabolism, and glucose availability through their augmentation, combating ischemic processes and oxidative stress, and decreasing inflammation (including with modulatory connotations, by the nuclear factor-κB pathway) and apoptosis-like processes, counteracting them by mitigating the caspase-3 activation induced by amyloid β-peptides. Since no available therapeutic agents are capable of curing the central nervous system's lesions, any contribution, such as that of Actovegin® (with consideration of a positive balance between benefits and risks), is worthy of further study and periodic reappraisal, including investigation into further connected aspects.


[Lesions of the peripheral nerves analyzed by classical electrodiagnosis].
In a sample of 102 patients 123 lesions of peripheral nerve due to injury were found. Diagnosis was not established in 27.0% cases. In course of three months after injuries 24.0% of patients were sent on electrodiagnostic procedures, the rest of 76.0% 4-14 months after peripheral nerve injury. 15.0% of patients were treated surgically in course of two months after injury and 85.0% of patients 4-16 months after injury. 32.0% were iatrogenic lesions, due to external fixator and extension, in field of distribution of n. radialis and n. fibularis. Complete lesion of peripheral nerve was found in 60.0%, and partial lesion of nerve in 37.0% cases, that is in correlation with results of surgery: 58.0% neuroraphias, 37.0% neurolysis and only 5.0% transplantation of nerves. All neuroraphias were secondary.


[Expression of CD20 in B-cell precursor acute lymphoblastic leukemia and its correlation with clinical outcomes].
To determine whether expression of CD20 is associated with clinical outcomes of childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL). 271 newly diagnosed childhood BCP-ALL during January 2009 to May 2013 were enrolled in this study. The patients were treated in line with the Chinese Childhood Leukemia Group ALL 2008 protocol (CCLG-ALL 2008). The clinical feature, early therapeutic response and clinical outcomes of the patients with a CD20 positive (CD20+ BCP) expression were compared with those with a CD20 negative (CD20- BCP) expression. CD20- BCP accounted for 45.76% (124 cases) of all participants. There were no significant differences between CD20- BCP and CD20- BCP patients in gender distribution, age, WBC counts when diagnosis was made, proportion of prednisone poor responders, and distribution of risk categories (P > 0.05). Patients of 10 years or older comprised 25.81% and 14.29% of CD20+ BCP and CD20- BCP patients, respectively (P = 0.017). Pro-B and pre-B cases accounted for 43.55% and 59.86% of CD20- BCP patients respectively, compared with 56.45 and 40.14% in CD20- BCP patients (P = 0.007). CD20+ BCP patients had 12.20% Philadelphia positive ALL and 6.50% BCP-ALL with TEL-AML1 fusion gene, compared with 4.86% (P = 0.03) and 18.06% (P = 0.005) in those of CD20 BCP. No significant differences were found between the two groups of patients in 15-day (77.50% vs. 74.13%, P = 0.525) and 33-day (95.04% vs. 95.83%, P = 0.757) complete remission rates. No significant differences (P > 0.05) were found in predicted 4-year event-free survival CEFS (78.00% +/- 4.96%) vs. (79.05% +/- 5.40%)) and predicted 4-year overall survival (OS (83.01% +/- 6.13%) vs. (93.64% +/- 2.46%)) between the two groups of patients either. CD20 positivity was not found to be associated with worse prognosis of children with BCP-ALL. More studies are needed to validate the correlation between CD20 and unfavorable outcomes in BCP-ALL.


Development of safe and efficacious viral vaccines for animals.
We have taken a number of approaches to improve the safety and efficacy of recombinant vaccines for use in humans and animals, including: choice of the strain of vaccinia virus (VACV) used as a vector, insertional inactivation of virulence and immunoregulatory genes of VACV, and expression of cytokine genes that attenuate the vector by more than a million-fold without reduction in immunogenicity. These strategies are illustrated by providing examples of recombinant VACV (rVACV) vaccines we have developed for rinderpest, vesicular stomatitis, simian immunodeficiency virus, and smallpox. We constructed rVACVs expressing interferon-gamma (IFN-gamma) and lacking the immune-modulating genes B8R, B13R, and B22R. IFN-gamma is a cytokine with potent immunoregulatory, antineoplastic, and antiviral properties. These rVACVs replicated to high titers in tissue culture, yet were avirulent in both immunocompromised and immunocompetent mice with no detectable viral replication in these animals. A single immunization elicited potent humoral, T-helper, and cytotoxic T-cell immune responses in mice despite the absence of any detectable virus replication in vivo. IFN-gamma co-expression and the inactivation of one or more VACV immune-modulating genes provide an optimized method for increasing the safety while maintaining the efficacy of rVACV vaccines for use in humans and animals.


Cation-coupled bicarbonate transporters.
Cation-coupled HCO3(-) transport was initially identified in the mid-1970s when pioneering studies showed that acid extrusion from cells is stimulated by CO2/HCO3(-) and associated with Na(+) and Cl(-) movement. The first Na(+)-coupled bicarbonate transporter (NCBT) was expression-cloned in the late 1990s. There are currently five mammalian NCBTs in the SLC4-family: the electrogenic Na,HCO3-cotransporters NBCe1 and NBCe2 (SLC4A4 and SLC4A5 gene products); the electroneutral Na,HCO3-cotransporter NBCn1 (SLC4A7 gene product); the Na(+)-driven Cl,HCO3-exchanger NDCBE (SLC4A8 gene product); and NBCn2/NCBE (SLC4A10 gene product), which has been characterized as an electroneutral Na,HCO3-cotransporter or a Na(+)-driven Cl,HCO3-exchanger. Despite the similarity in amino acid sequence and predicted structure among the NCBTs of the SLC4-family, they exhibit distinct differences in ion dependency, transport function, pharmacological properties, and interactions with other proteins. In epithelia, NCBTs are involved in transcellular movement of acid-base equivalents and intracellular pH control. In nonepithelial tissues, NCBTs contribute to intracellular pH regulation; and hence, they are crucial for diverse tissue functions including neuronal discharge, sensory neuron development, performance of the heart, and vascular tone regulation. The function and expression levels of the NCBTs are generally sensitive to intracellular and systemic pH. Animal models have revealed pathophysiological roles of the transporters in disease states including metabolic acidosis, hypertension, visual defects, and epileptic seizures. Studies are being conducted to understand the physiological consequences of genetic polymorphisms in the SLC4-members, which are associated with cancer, hypertension, and drug addiction. Here, we describe the current knowledge regarding the function, structure, and regulation of the mammalian cation-coupled HCO3(-) transporters of the SLC4-family.


Efficiency of Generation of Surface Acoustic Waves in Barium Strontium Titanate Thin Films.
The excitation of surface acoustic waves (SAWs) on the surface of the ferroelectric film [barium strontium titanate (BST)] located on the dielectric substrate (silicon) was studied theoretically. We found that the most effective SAW excitation takes place when spontaneous polarization occurs in the film plane and the wave propagates along the direction adjacent to the direction of the spontaneous polarization vector. Based on a nonlinear model of phase transitions in solid BST solutions, the dependency of the material constants of piezo-effect equations on the misfit strain with a fixed concentration was obtained numerically. The effect of various misfit strains on SAW characteristics was studied for the film located on single-crystal silicon. It was shown that the effectiveness of SAW excitation increases as the misfit strain nears the boundary of phase transition.


The harder they fall.
The past decade may well be remembered as the era of the high-flying, aggressive leader. Corner-office titans like Kenneth Lay, Dennis Kozlowski, and Bernard Ebbers graced the covers of business magazines. They captured the public's fascination with their bold business moves and charismatic sound bites. Then scandal set in, and the stars fell to earth. In this article, social psychologist Roderick M. Kramer asks an important question: Why do so many leaders--not just in business, but also in politics, religion, and the media--display remarkable adeptness and ability while courting power, only to engage in even more remarkable bouts of folly once that power has been secured? Kramer, who has spent most of his career researching how leaders get to the top, says there is something about the process of becoming a leader that changes people in profound ways. The systems through which we select our leaders force executives to sacrifice the attitudes and behaviors that are essential to their survival once they have reached the top. Society has learned to consider risk taking and rule breaking as markers of good leadership. As a result, CEOs and other leaders lack the modesty and prudence needed to cope with the rewards and trappings of power. They come to believe that normal limits don't apply to them and that they are entitled to any spoils they can seize. The leaders who do remain grounded--who get to the top and stay there--exhibit five common psychological and behavioral habits: They simplify their lives, remaining humble and "awfully ordinary." They shine a light on their weaknesses instead of trying to cover them up. They float trial balloons to uncover the truth and prepare for the unexpected. They sweat the small stuff. And they reflect more, not less.


Longitudinal map of transcriptome changes in the Lyme pathogen Borrelia burgdorferi during tick-borne transmission.
Borrelia burgdorferi (Bb), the causative agent of Lyme disease, adapts to vastly different environments as it cycles between tick vector and vertebrate host. During a tick bloodmeal, Bb alters its gene expression to prepare for vertebrate infection; however, the full range of transcriptional changes that occur over several days inside of the tick are technically challenging to capture. We developed an experimental approach to enrich Bb cells to longitudinally define their global transcriptomic landscape inside nymphal Ixodes scapularis ticks during a transmitting bloodmeal. We identified 192 Bb genes that substantially change expression over the course of the bloodmeal from 1 to 4 days after host attachment. The majority of upregulated genes encode proteins found at the cell envelope or proteins of unknown function, including 45 outer surface lipoproteins embedded in the unusual protein-rich coat of Bb. As these proteins may facilitate Bb interactions with the host, we utilized mass spectrometry to identify candidate tick proteins that physically associate with Bb. The Bb enrichment methodology along with the ex vivo Bb transcriptomes and candidate tick interacting proteins presented here provide a resource to facilitate investigations into key determinants of Bb priming and transmission during the tick stage of its unique transmission cycle.


Treatment planning for particle radiation therapy.
Fast neutrons beams from the new medically dedicated cyclotrons in the US have depth dose characteristics comparable to photon beams from a 6-MV linear accelerator, at best. Treatment planning will have specific difficulties related to the relatively increased radiosensitivities of the brain, spinal cord, lens of the eye, and salivary gland. Therefore, exploitation of the potential biologic advantages compared to high energy photons will extract the price of increased difficulties in treatment planning. Dosimetric advantages of protons and helium ions compared to high energy photons are real and make possible the high-dose irradiation of cancers immediately adjacent to sensitive critical normal structures. Treatment planning and delivery with a precision of less than 2 mm is necessary. Such methods are already operational. Particle radiation therapy facilities are national resources, which can help the clinical radiation oncologist in the unique management of a few, specific problems.


Mortality rates of hip fracture patients with non-operative treatment.
In this study, we aimed to analyze patient mortality rate after non-operative treatment of hip fractures to determine the distribution of causes of death and to compare factors affecting mortality. Between January 2013 and March 2019, a total of 93 patients (17 males, 76 females; mean age: 86.0±7.4 years; range, 64 to 98 years) who had hip fractures and were treated non-operatively were analyzed retrospectively. Survival, date of death, and cause of death were collected and analyzed. Baseline demographics, pre-trauma ambulation, pre- and post-trauma residence status, American Society of Anesthesiologists Physical Status (ASA PS) classification, and Parker's mobility score were compared with one-year mortality rates. The mean follow-up of survivors was 16.1±11.9 (range, 6.3 to 79.6) months. The mean survival of non-survivors was 4.9±6.1 (range, 0.007 to 27.3) months. The 3-, 6-, 12-, and 24-month mortality rates were 40.9%, 53.3%, 74.4%, and 87.5%, respectively. Respiratory diseases (33.3%) and cardiovascular diseases (13.6%) were the main causes of death among the patients. There was no statistically significant difference between the patients' age, sex, fracture site, pre-trauma ambulation, pre- and post-trauma residence status, ASA PS classification, Parker's mobility score, and one-year mortality. A significant number of patients are still treated non-operatively after hip fractures, and they have a high mortality rate. Efforts and research are needed to reduce mortality and improve the quality of life.


Acetate templating on calibrated standing digital radiograph improves accuracy of preoperative templating for total hip arthroplasty.
The accuracy of preoperative templating with respect to leg length, femoral offset, and the size of femoral and acetabular components is essential to the success of total hip arthroplasty (THA). Traditionally, templating has been performed using printed film with acetate templates. However, preoperative templating designed by different prosthetic manufacturers cannot be used directly on the film due to varying acetate template amplification ratios. Computer-based templating needs specialized digital templating software, which has cost implications. To address these shortcomings, we bring forward several questions: (1) the accuracy of traditional manual templating combined with the calibrated digital radiograph for preoperative templating, (2) the inter- and intraobserver reliability of this method. Using calibrated digital radiograph with traditional manual templating improves the accuracy and reproducibility of preoperative templating for THA. We designed a stepwise method that combines the traditional manual templating with standing digital radiograph calibrated by a scaling ball. Two separate observers (XJL, QYG) analyzed data of 82 patients (109 THAs) who had undergone THA with preoperative templating using the calibrated digital templating. The intra- and interobserver reliability was assessed by intraclass correlation coefficient. The size of the acetate template acetabular based on our method was identical to the actual implanted acetabular size in 55.0% (XJL 110/218 [50.5%]; QYG 130/218 [59.6%]) of the cases indicating moderate accuracy. The intraclass correlation coefficient (ICC) for acetabular templating indicated almost perfect interobserver (ICC=0.918 [95% CI, 0.893-0.937]) and intraobserver agreement (ICC=0.932 [95% CI, 0.912-0.947]). While the exact implanted femoral size was predicted in 55.3% (XJL 122/218 [56.0%]; QYG 119/218 [54.6%]) of the cases. The ICC for femoral component templating indicated almost perfect interobserver (ICC=0.944 [95% CI, 0.927-0.957]) and intraobserver agreement (ICC=0.909 [95% CI, 0.883-0.930]). This new stepwise method may prove to be a more reliable preoperative design choice to accurately calibrate magnification with radiograph, and could solve the incompatibility of the preoperative template designed by different prosthetic companies for direct use with the x-ray film. The method described is practical, convenient, cost-effective and does not require specialized equipment or software, thus making it particularly suitable for use in underdeveloped settings. Level IV, case series without controls.


Efficient Approach To Discover Novel Agrochemical Candidates: Intermediate Derivatization Method.
Intensive competition of intellectual property, easy development of agrochemical resistance, and stricter regulations of environmental concerns make the successful rate for agrochemical discovery extremely lower using traditional agrochemical discovery methods. Therefore, there is an urgent need to find a novel approach to guide agrochemical discovery with high efficiency to quickly keep pace with the changing market. On the basis of these situations, here we summarize the intermediate derivatization method (IDM) between conventional methods in agrochemicals and novel ones in pharmaceuticals. This method is relatively efficient with short time in discovery phase, reduced cost, especially good innovated structure, and better performance. In this paper, we summarize and illustrate "what is the IDM" and "why to use" and "how to use" it to accelerate the discovery of new biologically active molecules, focusing on agrochemicals. Furthermore, we display several research projects in our novel agrochemical discovery programs with improved success rate under guidance of this strategy in recent years.


Search for identical octapeptides in unrelated proteins: Structural plasticity revisited.
Since proteins are dynamic in nature, they can alter their local structure in response to changes in their environment factors such as temperature, pH, phosphorylation, and binding of other small molecules. These conformational changes are extremely important for the correct folding and functioning of proteins. There are also a number of diseases associated with protein conformational change such as amyloid diseases. To stimulate research into the above factors which specify one conformation over another, different theoretical models have been proposed and tested against sequence similar distant structure protein fragments. In order to simplify the computational complexity of identifying conformational changes in proteins, various local sequence search algorithms were employed and the structural plasticity in unrelated proteins was examined by various research groups. In the present work, we revisit the mechanism of structural plasticity in unrelated proteins with increased number of structures in Protein Data Bank by comparing identical octapeptides in unrelated proteins with dictionary of protein secondary structure extracted from existing experimental data. Our goal is to bring out the influence of hydrophobic residues, hydrophilic residues, flanking residues, difference in secondary structural propensities of surrounding residues, difference in phi-psi angles and local and nonlocal interactions in identical octapeptides adopting different conformations. Also we have used surrounding hydrophobicity, environment dependent interaction energy, atomic mean force potential, structural unit contacts and difference profiles models to explore the factors which cause structural plasticity. The results discussed here may provide insights into protein folding, design and function.


Research on recognition and intervention of behavior sequences in virtual museum learning.
Learning in virtual museum can transcend the limits of time and space. The virtual museum that combines expertise in different disciplines provides a virtual learning environment for college students, but how to intervene in museum learning has been unclear. Targeted at this question, this study selected 2030 majors in clinical medicine from a certain university and the final results exhibited four types of learners who are of high, medium, low and absent museum immersion, respectively. When the learners visited the virtual museum, their behavior data were collected backstage and later used as data source. The method of fuzzy c clustering analysis was utilized to test the behavior recognition results of virtual museum learning, and lag sequential analysis (LSA) was used to carry out sequential transformation of learning behaviors in virtual museum. In this study, the four types of learners were subsumed under two broad categories of middle & high museum immersion and low & absent museum immersion. The importance of behavior was identified with random forest algorithm, and the intervention mechanism of museum teaching was designed according to the analysis results. Specifically, such strategies as museum support, voice guidance, video guidance, sub-museum ordering, rewards points on the list, etc. were used to study the museum learners in need of intervention. The results showed that the learning state of some learners was significantly improved.


Debate on annual influenza vaccination in healthcare workers for the same subtype under the vaccine shortage.
Influenza vaccination is the primary method for preventing influenza and its severe complications. Healthcare workers (HCWs) are one of the priority groups for the influenza vaccination. To determine whether healthy HCWs, who were vaccinated with the same subtype for the two previous years, could be given less priority for influenza immunization under the vaccine shortage. We measured hemagglutination-inhibition antibody titers from sequential serum samples in 50 pre-immune subjects and 50 age-matched vaccine-naive subjects: immediately prior to the administration of the vaccine, 4-6 weeks, and 6 months after the vaccination. Prevaccination titers were maintained above protective level and high protection rates were observed for all three strains in pre-immune subjects: A/H1N1, A/H3N2, and B strains. As for the sequential changes, the protection rates for all three strains still remained above 70% until 6 months following the vaccination. Skipping influenza vaccination for a year could be considered in healthy pre-immune HCWs under the epidemic of the same subtype as two previous years.


In Vivo Assessment of Size-Selective Glomerular Sieving in Transplanted Human Induced Pluripotent Stem Cell-Derived Kidney Organoids.
The utility of kidney organoids in regenerative medicine will rely on the functionality of the glomerular and tubular structures in these tissues. Recent studies have demonstrated the vascularization and subsequent maturation of human pluripotent stem cell-derived kidney organoids after renal subcapsular transplantation. This raises the question of whether the glomeruli also become functional upon transplantation. We transplanted kidney organoids under the renal capsule of the left kidney in immunodeficient mice followed by the implantation of a titanium imaging window on top of the kidney organoid. To assess glomerular function in the transplanted human pluripotent stem cell-derived kidney tissue 1, 2, and 3 weeks after transplantation, we applied high-resolution intravital multiphoton imaging through the imaging window during intravenous infusion of fluorescently labeled low and high molecular mass dextran molecules or albumin. After vascularization, glomerular structures in the organoid displayed dextran and albumin size selectivity across their glomerular filtration barrier. We also observed evidence of proximal tubular dextran reuptake. Our results demonstrate that human pluripotent stem cell-derived glomeruli can develop an appropriate barrier function and discriminate between molecules of varying size. These characteristics together with tubular presence of low molecular mass dextran provide clear evidence of functional filtration. This approach to visualizing glomerular filtration function will be instrumental for translation of organoid technology for clinical applications as well as for disease modeling.


[Indicators monitoring the process of specialized nutritional support. Grupo de Nutrición de la SEFH].
To identify and develop monitoring indicators of the process of specialized nutritional support that will allow measuring the level of adherence to the established practice standards. Those practice standards considered to be key elements of the process were selected to develop performance indicators. The construction of these indicators combined the scientific evidence with expert opinion. Key goals were identified within each standard provided that its consecution would allow increasing the achievement of the standard. Particular improvement initiatives associated to each key goal were generated. Lastly, monitoring indicators were defined allowing undertaking a follow-up of the implementation of the improvement initiatives or either to assess the level of achievement of the key goals identified. Nineteen practice standards were selected representative of the critical points of the process. The strategic map for each standard has been defined, with the identification of 43 key goals. In order to achieve these key goals, a portfolio of improvements has been generated comprising 56 actions. Finally, 44 monitoring indicators have been defined grouped into three categories: 1. Numeric: they assess the level of goal achievement; 2. Dichotomic (yes/no): they inform on the execution of the improvement actions; 3. Results of the practice audits. We have made available monitoring indicators that allow assessing the level of adherence to the practice standards of the process of specialized nutritional support and the impact of the implementation of improvement actions within this process.


Functional Local Input to Layer 5 Pyramidal Neurons in the Rat Visual Cortex.
Pyramidal neurons in layer 5 of the neocortex can be differentiated into 3 cell subtypes: 1) short regular spiking (SH), 2) tall regular spiking (TR), and 3) tall burst spiking (TB), based on their morphological and electrophysiological properties. We characterized the functional excitatory local input to these 3 cell subtypes in rat primary visual cortex using laser-scanning photostimulation. Although all cell types received significant input from all cortical layers, SH neurons received stronger input from layer 4 and weaker input from layer 5 than did tall pyramidal cells. However, the laminar input to the 2 populations of tall pyramidal cells was indistinguishable. Simultaneous paired recording were then used to calculate a correlation probability (CP) to infer the proportion of shared input based on the occurrence of simultaneous synaptic potentials. Tall pairs of matched type had significantly higher CPs compared with unmatched pairs, suggesting that subpopulations of layer 4, 5, and 6 neurons preferentially connect to each tall cell type. Hence, this study shows that unconnected but matching pairs of tall pyramidal neurons, but not short pyramidal neurons, receive functional input from different interconnected networks within layers 4, 5, and 6.


Can Neck Contusions in Unexplained Pediatric Deaths be Explained by Cardiopulmonary Resuscitation?
The distribution of cutaneous contusions in infants may raise the possibility of maltreatment. Neck contusions are particularly problematic since they seldom occur outside the setting of abuse, while cardiopulmonary resuscitation (CPR)-related maneuvers may involve the neck. To address the role of CPR in neck findings, we examined 260 consecutive pediatric autopsies in which CPR was attempted. No neck contusions were identified in manners of death classified as natural, undetermined, or suicide. Contusions were present in two of 80 deaths classified as accident and had obvious accidental causes. About 26% of deaths classified as homicide had neck contusions with no explanation provided by the caregiver (p < 0.0001). In conclusion, neck contusions in deceased children with no apparent explanation should be regarded as suspicious for abuse and investigated accordingly. CPR is not a plausible explanation for neck contusions in children.


Comparison of different fluorimetric HPLC methods for analysis of acidic polyether toxins in marine phytoplankton.
The human toxic syndrome, diarrhetic shellfish poisoning (DSP), is caused by polyether toxins that are present in bivalve molluscs but originate from some species of marine phytoplankton. During the last few years different HPLC methods with fluorescence detection (FLD) have been proposed for analysis of marine toxins, including polyether toxins, in shellfish and phytoplankton. Several derivatization reagents have been proposed in the literature, with the aim of converting the acidic DSP toxins into their corresponding fluorescent derivatives. In this work we report results obtained from HPLC-FLD analysis of extracts from phytoplankton, including Dinophysis spp.,harvested off the south-west coast of Ireland. Three different reagents were used for fluorescent derivatization: 3-bromomethyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone (BrDMEQ), 9-chloromethylanthracene (CA), and "in situ" 9-anthracenyldiazomethane (ADAM). Derivatization was performed under conditions previously optimised. The DSP derivatives were cleaned using different SPE procedures then analysed by HPLC-FLD. In this study, the use of BrDMEQ, CA, and "in situ" ADAM was compared in terms of sensitivity and selectivity. Evaluation of HPLC methods for analysis of DSP toxin derivatives was also conducted; the presence of okadaic acid (OA), dinophysistoxin-2 (DTX-2), and pectenotoxin-2 seco acids (PTX1SAs) was detected in the sample extracts studied.


Nonossifying fibromyxoid tumor of soft tissue.
A firm, painless tumor of the temporal region was excised in a 3-year-old girl. The diagnosis of nonossifying fibromyxoid tumor was established on pathologic examination, which showed that most of the cells formed cords and tubulelike structures in a myxoid background. Bone and osteoid were lacking. The child is free of recurrence 4 years after surgery.