[Infantile neuroaxonal dystrophy: diagnosis during life by biopsy of the conjunctiva].
Three cases of infantile neuroaxonal dystrophy diagnosed by conjunctival biopsy are reported. Some axons of the conjunctival nerves showed aggregates of tubular and membranous structures identical to the spheroids of the CNS. The visualization of these structures is the only diagnostic tool in this disease of unknown metabolic basis. Conjunctival biopsy which is ease to perform is the preferential technique for the diagnosis of this disease. Clinically, the intense hypotonia with pyramidal tract signs, the absence of seizures, the cerebellar atrophy observed at the computerized tomography suggest strongly the diagnosis of infantile neuroaxonal dystrophy.


Veronica linariifolia subsp. dilatata ameliorates LPS-induced acute lung injury by attenuating endothelial cell barrier dysfunction via EGFR/Akt/ZO-1 pathway.
The dried aerial parts of Veronica linariifolia subsp. dilatata (Nakai & Kitag.) D.Y.Hong named Shui Man Jing (SMJ) is a traditional Chinese medicine with a long history of clinical use in the treatment of chronic bronchitis and coughing up blood, however, its role on acute lung injury (ALI) has not been revealed yet. To assess the efficiency of SMJ on ALI and to investigate whether it inhibited endothelial barrier dysfunction by regulating the EGFR/Akt/ZO-1 pathway to alleviate ALI in vivo and in vitro based on the result of network pharmacology. An in vivo model of ALI was established using inhalation of atomized lipopolysaccharide (LPS), and the effects of SMJ on ALI were evaluated through histopathological examination and inflammatory cytokines, lung histology and edema, vascular and alveolar barrier disruption. Network pharmacology was applied to predict the mechanism of SMJ in the treatment of ALI. The crucial targets were validated by RT-PCR, Western Blotting, molecular docking, immunohistochemistry and immunofluorescence methods in vivo and in virto. Administration of SMJ protected mice against LPS-induced ALI, including ameliorating the histological alterations in the lung tissues, and decreasing lung edema, protein content of bronchoalveolar lavage fluid, infiltration of inflammatory cell and secretion of cytokines. SMJ exerted protective effects in ALI by inhibiting endothelial barrier dysfunction in mice and bEnd.3 cell. SMJ relieved endothelial barrier dysfunction induced by LPS through upregulating the EGFR expression. SMJ also increased the phosphorylation of Akt, and ZO-1 expression both in vivo and in vitro. SMJ attenuates vascular endothelial barrier dysfunction for LPS-induced ALI via EGFR/Akt/ZO-1 pathway, and is a promising novel therapeutic candidate for ALI.


Discrepancies in the recovery of bacteria from multiple sinuses in acute and chronic sinusitis.
The microbiology of acute and chronic sinusitis has been studied extensively. Establishing the concomitant distribution of the causative organisms in cases that involve multiple sinuses is of scientific and practical importance. This study evaluated the aerobic and anaerobic microbiology of acute and chronic sinusitis in patients with involvement of multiple sinuses. The 155 patients evaluated had sinusitis of either the maxillary, ethmoid or frontal sinuses (any combination) and had organisms recovered from two to four concomitantly infected sinuses. Similar aerobic, facultatively anaerobic and anaerobic organisms were recovered from all groups of patients. In patients who had organisms isolated from two sinuses and had acute sinusitis, 31 (56 %) of the 55 isolates were found only in a single sinus, and 24 (44 %) were recovered concomitantly from two sinuses. In those with chronic infection 31 (34 %) of the 91 isolates were recovered only from a single sinus, and 60 (66 %) were found concomitantly from two sinuses. Anaerobic bacteria were more often isolated concomitantly from two sinuses (50 of 70) than aerobic and facultatively anaerobic (ten of 21, P < 0.05). Similar findings were observed in patients who had organisms isolated from three or four sinuses. beta-Lactamase-producing bacteria were more often isolated from patients with chronic infection (58-83 %) as compared to those with acute infections (32-43 %). These findings illustrate that there are differences in the distribution of organisms in single patients who suffer from infections in multiple sinuses and emphasize the importance of obtaining cultures from all infected sinuses.


Fludarabine and Melphalan Compared with Reduced Doses of Busulfan and Fludarabine Improve Transplantation Outcomes in Older Patients with Myelodysplastic Syndromes.
Reduced-intensity conditioning (RIC) regimens developed to extend the use of allogeneic hematopoietic stem cell transplantation (HSCT) to older patients have resulted in encouraging outcomes. We aimed to compare the 2 most commonly used RIC regimens, i.v. fludarabine with busulfan (FluBu) and fludarabine with melphalan (FluMel), in patients with myelodysplastic syndrome (MDS). Through the Center for International Blood and Marrow Transplant Research (CIBMTR), we identified 1045 MDS patients age ≥60 years who underwent first HSCT with a matched related or matched (8/8) unrelated donor using an RIC regimen. The CIBMTR's definition of RIC was used: a regimen that incorporated an i.v. busulfan total dose ≤7.2 mg/kg or a low-dose melphalan total dose ≤150 mg/m2. The 2 groups, recipients of FluBu (n = 697) and recipients of FluMel (n = 448), were comparable in terms of disease- and transplantation-related characteristics except for the more frequent use of antithymocyte globulin or alemtuzumab in the FluBu group (39% versus 31%). The median age was 67 years in both groups. FluMel was associated with a reduced relapse incidence (RI) compared with FluBu, with a 1-year adjusted incidence of 26% versus 44% (P ≤ .0001). Transplantation-related mortality (TRM) was higher in the FluMel group (26% versus 16%; P ≤ .0001). Because the magnitude of improvement with FluMel in RI was greater than the improvement in TRM with FluBu, disease-free survival (DFS) was better at 1 year and beyond with FluMel compared with FluBu (48% versus 40% at 1 year [P = .02] and 35% versus 27% at 3 years [P = .01]). Overall survival was comparable in the 2 groups at 1 year (63% versus 61%; P = .4) but was significantly improved with FluMel compared with FluBu at 3 years (46% versus 39%; P = .03). Our results suggest that FluMel is associated with superior DFS compared with FluBu owing to reduced RI in older patients with MDS patients. © 2021 American Society for Transplantation and Cellular Therapy. Published by Elsevier Inc.


BioLiP2: an updated structure database for biologically relevant ligand-protein interactions.
With the progress of structural biology, the Protein Data Bank (PDB) has witnessed rapid accumulation of experimentally solved protein structures. Since many structures are determined with purification and crystallization additives that are unrelated to a protein's in vivo function, it is nontrivial to identify the subset of protein-ligand interactions that are biologically relevant. We developed the BioLiP2 database (https://zhanggroup.org/BioLiP) to extract biologically relevant protein-ligand interactions from the PDB database. BioLiP2 assesses the functional relevance of the ligands by geometric rules and experimental literature validations. The ligand binding information is further enriched with other function annotations, including Enzyme Commission numbers, Gene Ontology terms, catalytic sites, and binding affinities collected from other databases and a manual literature survey. Compared to its predecessor BioLiP, BioLiP2 offers significantly greater coverage of nucleic acid-protein interactions, and interactions involving large complexes that are unavailable in PDB format. BioLiP2 also integrates cutting-edge structural alignment algorithms with state-of-the-art structure prediction techniques, which for the first time enables composite protein structure and sequence-based searching and significantly enhances the usefulness of the database in structure-based function annotations. With these new developments, BioLiP2 will continue to be an important and comprehensive database for docking, virtual screening, and structure-based protein function analyses.


Targeting PKC alleviates iron overload in diabetes and hemochromatosis.
Diabetes is one of the most prevalent chronic diseases worldwide. Iron overload increases the incidence of diabetes and aggravates diabetic complications that cause mortality. Reciprocally, diabetes potentially promotes body iron loading, but the mechanism remains not well understood. In this study, we demonstrated systemic iron excess and the upregulation of iron exporter ferroportin (Fpn) in the enterocytes and macrophages of multiple diabetic mouse models. Increased Fpn expression and iron efflux was also seen in the enterocytes of type 2 diabetic human patients. We further showed that protein kinase C (PKC), which is activated in hyperglycemia, was responsible for the sustained membrane expression of Fpn in physiological and in diabetic settings. For the first time, we identified that PKCs were novel binding proteins and positive regulators of Fpn. Mechanistically, hyperactive PKC promoted exocytotic membrane insertion while inhibited the endocytic trafficking of Fpn in the resting state. PKC also protected Fpn from internalization and degradation by its ligand hepcidin dependent on decreased ubiquitination and increased phosphorylation of Fpn. Importantly, the loss-of-function and pharmacological inhibition of PKC alleviated systemic iron overload in diabetes and hemochromatosis. Our study thus highlights PKC as a novel target in the control of systemic iron homeostasis.


Comparison of standardbred trotters exercising on a treadmill and a race track with identical draught resistances.
The responses in heart rate, plasma lactate and rectal temperature of standardbred trotters to draught loaded interval exercise on a treadmill and a race track were studied. The horses were exercised with incrementally increasing trotting speeds for two-minute intervals with draught loads of 10, 20 and 30 kilopond (kp) in three different tests. Each trotting interval was followed by two-minute periods at a walk without a draught load. Measurements of heart rate and plasma lactate were made at the end of each interval and the rectal temperature was taken at the end of the exercise. The heart rate and plasma lactate levels were significantly lower on the treadmill than on the track in the tests with 10 kp, but no significant differences were found between the treadmill and track exercise tests with the heavier draught resistances. No differences were observed in rectal temperature between treadmill and track conditions. From these findings it was concluded that the workload was significantly greater on the race track compared to the treadmill when the draught resistance was low (10 kp). Although the workload increased on both the race track and the treadmill as draught resistance increased, at the heavier draught resistances track exercise was no longer more demanding than exercise on the treadmill.


A Soft Skin Adhesive (SSA) Patch for Extended Release of Pirfenidone in Burn Wounds.
As much as half or more of deep partial-thickness burn wounds develop hypertrophic scarring and contracture. Once formed, treatments are only minimally effective. Pirfenidone (Pf), indicated for treatment of idiopathic pulmonary fibrosis, is an anti-inflammatory and anti-fibrotic small molecule that potentially can be repurposed as a preventative against scarring in burn wounds. We present a drug-in-matrix patch with a soft skin adhesive (SSA) wound-contacting layer for multi-day drug delivery of Pf into burn wounds at the point of injury. Our patch construction consists of an SSA adhesive layer (Liveo™ MG7-9850, Dupont, Wilmington, DE, USA) for wound fixation, an acrylic co-polymer drug matrix (DURO-TAK 87-2852, Henkel, Düsseldorf, Germany) as the drug (Pf) reservoir, and an outermost protective polyurethane backing. By employing a drug-in-matrix patch design, Pf can be loaded as high as 2 mg/cm2. Compared to the acrylic co-polymer adhesive patch preparations and commercial films, adding an SSA layer markedly reduces skin stripping observed under scanning electron microscopy (SEM). Moreover, the addition of varying SSA thicknesses did not interfere with the in vitro release kinetics or drug permeation in ex vivo porcine skin. The Pf patch can be easily applied onto and removed from deep partial-thickness burn wounds on Duroc pigs. Continuous multi-day dosing of Pf by the patches (>200 μg/cm2/day) reduced proinflammatory biomarkers in porcine burn wounds. Pf patches produced by the manual laboratory-scale process showed excellent stability, maintaining intact physical patch properties and in vitro biological activity for up to one year under long-term (25 °C at 60% RH) and 6 months under accelerated (40 °C at 75% RH) test conditions. To manufacture our wound safe-and-extended-release patch, we present scale-up processes using a machine-driven automated roll-to-roll pilot scale coater.


Mucosal malignant melanoma - a clinical, oncological, pathological and genetic survey.
Mucosal melanomas constitute 1.3% of all melanomas and they may develop in any mucosal membrane. Conjunctival melanomas (0.5/million/year) and melanomas in the sinonasal cavity (0.5/million/year) are the most common, followed by anorectal melanomas (0.4/million/year) and melanomas in the oral cavity (0.2/million/year). Anorectal melanoma occurs slightly more often in females, whereas oral melanoma has a male predilection. Mucosal melanoma most commonly develops in a patient's sixth or seventh decade of life, and no differences between races have been found except for sinonasal melanoma and conjunctival melanoma, which are very rare in Black people. The symptoms are not tumour-specific and are related to the organ system affected, and the disease is most often diagnosed at an advanced clinical stage. The diagnosis of a primary tumour is difficult, and metastatic cutaneous melanoma and choroidal melanoma must be excluded. Mutations in KIT are frequently found, while BRAF and NRAS mutations are rarely found - except in conjunctival melanomas that carry BRAF mutations. Mutations in the TERT promotor region are also found in mucosal melanomas. Complete surgical resection with free margins is the treatment of choice. The prognosis is poor, with the 5-year survival rate ranging from 0% (gastric melanoma) to 80% (conjunctival melanoma).


Photochemistry of glyoxylate embedded in sodium chloride clusters, a laboratory model for tropospheric sea-salt aerosols.
Although marine aerosols undergo extensive photochemical processing in the troposphere, a molecular level understanding of the elementary steps involved in these complex reaction sequences is still missing. As a defined laboratory model system, the photodissociation of sea salt clusters doped with glyoxylate, [NanCln-2(C2HO3)]+, n = 5-11, is studied by a combination of mass spectrometry, laser spectroscopy and ab initio calculations. Glyoxylate acts as a chromophore, absorbing light below 400 nm via two absorption bands centered at about 346 and 231 nm. Cluster fragmentation dominates, which corresponds to internal conversion of the excited state energy into vibrational modes of the electronic ground state and subsequent unimolecular dissociation. Photochemical dissociation pathways in electronically excited states include CO and HCO elimination, leading to [Nan-xCln-x-2HCOO]+ and [NanCln-2COO˙]+ with typical quantum yields in the range of 1-3% and 5-10%, respectively, for n = 5. The latter species contains CO2˙- stabilized by the salt environment. The comparison of different cluster sizes shows that the fragments containing a carbon dioxide radical anion appear in a broad spectral region of 310-380 nm. This suggests that the elusive CO2˙- species may be formed by natural processes in the troposphere. Based on the photochemical cross sections obtained here, the photolysis lifetime of glyoxylate in a dry marine aerosol is estimated as 10 h. Quantum chemical calculations show that dissociation along the C-C bond in glyoxylic acid as well as glyoxylate embedded in the salt cluster occurs after reaching the S1/S0 conical intersection, while this conical intersection is absent in free glyoxylate ions.


Precise reporting of traditional Chinese medicine interventions in randomized controlled trials.
Traditional Chinese medicine (TCM) intervention should be concisely and precisely reported in randomized controlled trials (RCTs). Based on State Food and Drug Administration's categories, we recommend reporting the interventions as follows: (1) Single Chinese herbal medicine-based/formula-based/extraction-based intervention includes 1) Name, dosage format and registration; 2) The composition and quality of intervention; 3) Pharmaceutical processing and quality control; 4) Stability of final product and quality control; 5) Function and safety description; 6) Dosage and treatment course; 7) Control group. (2) Active compound-based TCM drug intervention includes 1) Name of active compound(s); 2) Original source of active compound(s); 3) The brief process obtaining active compound(s); 4) Percentage of active compound(s) in final product; 5) Added materials and its quality and quantity control. Besides, the detailed information of intervention can be published as an online supplement in web site.


Association of sex of fetus, sire, month of conception, or year of foaling with duration of gestation in standardbred mares.
To determine whether sex of fetus, sire, month of conception, or year of foaling was associated with duration of gestation in mares. Epidemiologic retrospective cohort study. 500 foalings for 296 Standardbred mares. Data for reproductive events from 1986 to 1992 were analyzed. Analyses were conducted to determine whether duration of gestation was associated with sex of fetus, sire, month of conception, or year of foaling. Mean duration of gestation was 343.3 days and was significantly greater for colt fetuses (344.4 days) than for filly fetuses (342.2 days). Sire was associated with duration of gestation; gestation after mating with certain sires was consistently less than 340 days in duration, whereas duration after mating with other sires was consistently more than 350 days. Duration of gestation was associated with month of conception, decreasing by about 2.5 d/mo for mares conceiving later during the breeding season. Duration of gestation is affected by mating to specific sires and by month of conception. Stallions associated with exceptionally prolonged gestations might be used to breed mares early in the breeding season, whereas stallions associated with shorter gestations might be desirable for mares bred later in the breeding season. Preparations for impending parturition should be more effective and efficient with improved information on expected date of parturition.


Immune profiling of plasma-derived extracellular vesicles identifies Parkinson disease.
To develop a diagnostic model based on plasma-derived extracellular vesicle (EV) subpopulations in Parkinson disease (PD) and atypical parkinsonism (AP), we applied an innovative flow cytometric multiplex bead-based platform. Plasma-derived EVs were isolated from PD, matched healthy controls, multiple system atrophy (MSA), and AP with tauopathies (AP-Tau). The expression levels of 37 EV surface markers were measured by flow cytometry and correlated with clinical scales. A diagnostic model based on EV surface markers expression was built via supervised machine learning algorithms and validated in an external cohort. Distinctive pools of EV surface markers related to inflammatory and immune cells stratified patients according to the clinical diagnosis. PD and MSA displayed a greater pool of overexpressed immune markers, suggesting a different immune dysregulation in PD and MSA vs AP-Tau. The receiver operating characteristic curve analysis of a compound EV marker showed optimal diagnostic performance for PD (area under the curve [AUC] 0.908; sensitivity 96.3%, specificity 78.9%) and MSA (AUC 0.974; sensitivity 100%, specificity 94.7%) and good accuracy for AP-Tau (AUC 0.718; sensitivity 77.8%, specificity 89.5%). A diagnostic model based on EV marker expression correctly classified 88.9% of patients with reliable diagnostic performance after internal and external validations. Immune profiling of plasmatic EVs represents a crucial step toward the identification of biomarkers of disease for PD and AP.


The Virulence of Different Vaccinia Virus Strains Is Directly Proportional to Their Ability To Downmodulate Specific Cell-Mediated Immune Compartments In Vivo.
Vaccinia virus (VACV) is a notorious virus for a number of scientific reasons; however, most of its notoriety comes from the fact that it was used as a vaccine against smallpox, being ultimately responsible for the eradication of that disease. Nonetheless, many different vaccinia virus strains have been obtained over the years; some are suitable to be used as vaccines, whereas others are virulent and unsuitable for this purpose. Interestingly, different vaccinia virus strains elicit different immune responses in vivo, and this is a direct result of the genomic differences among strains. In order to evaluate the net result of virus-encoded immune evasion strategies of vaccinia viruses, we compared antiviral immune responses in mice intranasally infected by the highly attenuated and nonreplicative MVA strain, the attenuated and replicative Lister strain, or the virulent WR strain. Overall, cell responses elicited upon WR infections are downmodulated compared to those elicited by MVA and Lister infections, especially in determined cell compartments such as macrophages/monocytes and CD4+ T cells. CD4+ T cells are not only diminished in WR-infected mice but also less activated, as evaluated by the expression of costimulatory molecules such as CD25, CD212, and CD28 and by the production of cytokines, including tumor necrosis factor alpha (TNF-α), gamma interferon (IFN-γ), interleukin-4 (IL-4), and IL-10. On the other hand, MVA infections are able to induce strong T-cell responses in mice, whereas Lister infections consistently induced responses that were intermediary between those induced by WR and MVA. Together, our results support a model in which the virulence of a VACV strain is proportional to its potential to downmodulate the host's immune responses.IMPORTANCE Vaccinia virus was used as vaccine against smallpox and was instrumental in the successful eradication of that disease. Although smallpox vaccination is no longer in place in the overall population, the use of vaccinia virus in the development of viral vector-based vaccines has become popular. Nonetheless, different vaccinia virus strains are known and induce different immune responses. To look into this, we compared immune responses triggered by mouse infections with the nonreplicative MVA strain, the attenuated Lister strain, or the virulent WR strain. We observed that the WR strain was capable of downmodulating mouse cell responses, whereas the highly attenuated MVA strain induced high levels of cell-mediated immunity. Infections by the intermediately attenuated Lister strain induced cell responses that were intermediary between those induced by WR and MVA. We propose that the virulence of a vaccinia virus strain is directly proportional to its ability to downmodulate specific compartments of antiviral cell responses.


Barriers and facilitators to the implementation of nurse's role in primary care settings: an integrative review.
The rapid evolution of the epidemiological picture and the recent SARS-COV-2 pandemic has expressed the vulnerabilities of health systems and focuses attention on the population's needs. The nurse's figure in the care teams is universally identified; however, the implementation of the role within some care settings turns out to be complex and challenging. This integrative review aims to identify the barriers and facilitators in implementing the role of the nurse in primary care settings. An integrative review was conducted on the Medline and Cinahl databases until 9 June 2020. Qualitative, quantitative, and Mixed-method research studies were selected to identify studies related to the barriers and facilitators of the nurse's role in nursing facilities' primary care. For the extraction of the results, the Consolidating Framework for Research Implementation (CFIR) was used to identify the factors that influence implementation in health care. Following the duplicates' removal, the search identified 18,257 articles, of which 56 were relevant to the inclusion criteria; therefore, they were included in the summary. The selected studies were conducted in thirteen countries, most from Oceania, Europe, North America, Latin America, and the Caribbean. The barriers reported most frequently concern the nursing profession's regulatory and regulatory aspects within the contexts of care, cultural and organizational aspects, training, and the transfer of specific skills, which were previously designated to doctors. The facilitators are mainly linked to the nurse's adaptability to the various contexts of care, recognizing the patient's role, and the desire to develop multidisciplinary and effective working groups to respond to the health needs of the population in primary care contexts. This review highlighted the main barriers and facilitators in implementing the nurse's role in primary care settings. These results offer useful elements for stakeholders to identify effective strategies in preparing programs and activities for implementing the nurse's role, acting on the elements identified as barriers and favouring the aspects that emerge as facilitators.


Electronic and magnetic properties of endohedrally doped fullerene Mn@C(60): a total energy study.
We perform total energy calculations on a manganese atom encapsulated inside a C(60) cage using density functional theory with the generalized gradient approximation through three optimization schemes and along four paths inside the cage. We find that when Mn is located in the central region, its electronic and magnetic properties are not exactly the same as those of a free Mn atom due to weak coupling between Mn and the cage. As Mn is shifted toward to the edge, the total energy and spin start to change significantly when Mn is situated about one-third of the way between the cage center and edge, and the total energy reaches a local minimum. Finally the interaction between Mn and the cage turns repulsive as Mn approaches the edge. We also find that, along the lowest energy path, there exist three consecutive local energy minima and each of these has a different spin M. The ground state has the lowest M=3, Mn is located about 1.6 A away from the cage center, and the binding energy is 0.08 eV. We attribute the decrease in total energy and spin to Mn and C hybridization.


Predicting breastfeeding in women living in areas of economic hardship: Explanatory role of the theory of planned behaviour.
This study employed the theory of planned behaviour (TPB) and additional variables (descriptive norm, moral norm, self-identity) to investigate the factors underlying breastfeeding intention and subsequent breastfeeding at four time points (during hospital stay, at hospital discharge, 10 days postpartum and 6 weeks postpartum) in a sample of women selected from defined areas of economic hardship (N = 248). A model containing the TPB, additional variables and demographic factors provided a good prediction of both intention (R (2) = 0.72; attitude, perceived behavioural control, moral norm and self-identity significant predictors) and behaviour - breastfeeding at birth (88.6% correctly classified; household deprivation, intention, attitude significant), at discharge from hospital (87.3% correctly classified; intention, attitude significant), 10 days after discharge (83.1% correctly classified; education, intention, attitude, descriptive norm significant) and 6 weeks after discharge (78.0% correctly classified; age, household deprivation, ethnicity, moral norm significant). Implications for interventions are discussed, such as the potential usefulness of targeting descriptive norms, moral norms and perceived behavioural control (PBC) when attempting to increase breastfeeding uptake.


[Historical retrospective of nursing research in Brazil].
Types and lines of nursing research in Brazil, from 1950 to 1980, are described. The majority of studies are on nursing care, followed by nursing education. The Author concludes that probably these studies were an answer to the needs of the country.


Statics and dynamics of free and hydrogen-bonded OH groups at the air/water interface.
We use classical atomistic molecular dynamics simulations of two water models (SPC/E and TIP4P/2005) to investigate the orientation and reorientation dynamics of two subpopulations of OH groups belonging to water molecules at the air/water interface at 300 K: those OH groups that donate a hydrogen bond (called "bonded") and those that do not (called "free"). Free interfacial OH groups reorient in two distinct regimes: a fast regime from 0 to 1 ps and a slow regime thereafter. Qualitatively similar behavior was reported by others for free OH groups near extended hydrophobic surfaces. In contrast, the net reorientation of bonded OH groups occurs at a rate similar to that of bulk water. This similarity in reorientation rate results from compensation of two effects: decreasing frequency of hydrogen-bond breaking/formation (i.e., hydrogen-bond exchange) and faster rotation of intact hydrogen bonds. Both changes result from the decrease in density at the air/water interface relative to the bulk. Interestingly, because of the presence of capillary waves, the slowdown of hydrogen-bond exchange is significantly smaller than that reported for water near extended hydrophobic surfaces, but it is almost identical to that reported for water near small hydrophobic solutes. In this sense water at the air/water interface has characteristics of water of hydration of both small and extended hydrophobic solutes.


The design and methodology of premature ejaculation interventional studies.
Large well-designed clinical efficacy and safety randomized clinical trials (RCTs) are required to achieve regulatory approval of new drug treatments. The objective of this article is to make recommendations for the criteria for defining and selecting the clinical trial study population, design and efficacy outcomes measures which comprise ideal premature ejaculation (PE) interventional trial methodology. Data on clinical trial design, epidemiology, definitions, dimensions and psychological impact of PE was reviewed, critiqued and incorporated into a series of recommendations for standardisation of PE clinical trial design, outcome measures and reporting using the principles of evidence based medicine. Data from PE interventional studies are only reliable, interpretable and capable of being generalised to patients with PE, when study populations are defined by the International Society for Sexual Medicine (ISSM) multivariate definition of PE. PE intervention trials should employ a double-blind RCT methodology and include placebo control, active standard drug control, and/or dose comparison trials. Ejaculatory latency time (ELT) and subject/partner outcome measures of control, personal/partner/relationship distress and other study-specific outcome measures should be used as outcome measures. There is currently no published literature which identifies a clinically significant threshold response to intervention. The ISSM definition of PE reflects the contemporary understanding of PE and represents the state-of-the-art multi-dimensional definition of PE and is recommended as the basis of diagnosis of PE for all PE clinical trials.


Five-year experience with a suture annuloplasty for mitral valve repair.
We present five years' experience with mitral plication annuloplasty, performed with a semicircular buttressed suture around the posterior leaflet in 130 patients (mean age 58 +/- 11 years) with primary mitral valve disease (n = 71) or functional mitral regurgitation (n = 59). In 65 cases the mitral valve itself was also repaired. Concomitant myocardial revascularization was performed in 40 cases and aortic valve replacement in 43. All but three patients were followed up (97.6%). Postoperative echocardiography showed acceptable mitral area (2.28 +/- 0.39 cm2) and good valve competence in all cases. Inhospital mortality was 3% and late mortality 4.8%. During the follow-up period (22.8 +/- 10.9 months) 8 patients (6.6%) required mitral valve replacement because of progression of native valve disease (n = 4), technical failure (2) or expansion of the annuloplasty suture (2). Mitral annuloplasty thus can be performed simply and with good results, using a strong, non-stretchable buttressed suture. This procedure can be an inexpensive alternative to ring implantation.


Aberrant retention of tyrosinase in the endoplasmic reticulum mediates accelerated degradation of the enzyme and contributes to the dedifferentiated phenotype of amelanotic melanoma cells.
The loss of tyrosinase, the key enzyme in melanin synthesis, has been implicated in the dedifferentiation of malignant melanocytes. The presence of tyrosinase transcripts and antigenic peptides in melanoma tumors prompted us to investigate whether the basis for the loss of the enzyme was proteolytic degradation. Toward this aim, we followed the kinetics of synthesis, degradation, processing, chaperone binding, inhibitor sensitivity, and subcellular localization of tyrosinase in normal and malignant melanocytes. We found that, in amelanotic melanoma cell lines, tyrosinase failed to reach the melanosome, the organelle for melanin synthesis, because it was retained in the endoplasmic reticulum (ER) and then degraded. Tyrosinase appeared mostly as a 70-kDa core-glycosylated, endoglycosidase H-sensitive, immature form bound to the ER chaperone calnexin and had a life-span of only 25% of normal. Maturation and transit from the ER to the Golgi compartment was facilitated by lowering the temperature of incubation to 31 degrees C. Several proteasome inhibitors caused the accumulation of an approximately 60-kDa tyrosinase doublet that was more prominent in malignant than in normal melanocytes and promoted, to various degrees, the maturation of tyrosinase in melanoma cells and the translocation of the enzyme to melanosomes. The appearance of ubiquitinated tyrosinase after treatment of normal melanocytes with N-acetyl-L-leucinyl-L-leucinal-L-norleucinal reinforced our notion that some tyrosinase is normally degraded by proteasomes. Proteolysis of tyrosinase by proteasomes is consistent with the production of antigenic tyrosinase peptides that are presented to the immune system by major histocompatibility complex class I molecules.


Late prasugrel benefit in STEMI patients?
Recent post hoc analysis of the TRITON trial suggests vascular outcome superiority of prasugrel over clopidogrel in STEMI patients at both 30 days and 15 months. However, this paper is not in full agreement with the data presented in the Food and Drug Administration (FDA) issued Prasugrel Second Review. While the periprocedural advantage of the experimental drug is confirmed by the independent FDA review, the late benefit is grossly exaggerated. The FDA document suggests that the benefit of prasugrel is exclusively front-loaded, with no additional long-term vascular advantage beyond one month if site-reported events are counted. It is unclear why secondary occlusive events in STEMI patients were consistently misdiagnosed by the TRITON investigators, especially considering that site reported MI's correlated with mortality, while adjudicated MI's exhibited similar to MI-free patients mortality rates. Inflation of the outcome superiority is especially alarming considering lack of heart failure benefit for prasugrel, challenging the hypothesis of delayed myocardial preservation by early prevention of "microMI's", or "saving the muscle" hypothesis. In contrast, significant steady growth over time of definite serious bleeding events, and unexpected cancer risks, especially in women, and probably delayed mortality risks with prasugrel are real, clearly dominate in the outpatient setting, and represent a valid safety concern.


Wet combing for head lice: feasibility in mass screening, treatment preference and outcome.
There is no scientific consensus on the best way to control head louse infestation in schoolchildren. A study was conducted to test the feasibility and acceptability of a screening campaign by wet combing and a community approach to head-louse control with home visits, and to explore parents' treatment preferences and treatment outcomes. A non-controlled intervention (advice on treatment options offered to all positive children) was nested within an epidemiological prevalence study. All children in three primary schools in Ghent, Belgium, were invited to take part in screening by wet combing (n=677, 3-11 years). Positive children were offered structural treatment advice, a home visit on day 7, and a check by wet combing on day 14. 83% of the children were screened. The prevalence of active infestation (living moving lice) was 13.0% in school 1 and 19.5% in school 3. In school 2, prevalence of signs of active and past infestation was 40.7%. A home visit was made to 58% of the positive children. 85% of the positive children were screened again on day 14. Wet combing was the most widely used treatment, followed by chemical treatment and a combination of the two. In school 1 and 3 51% were cured, and in school 2 24% became nit-free. A wet combing screening campaign and a community-oriented approach to head-louse control is feasible though resource-intensive. The prevalence of head lice was high and the cure rate was low, with either topical treatments or wet combing.


Effects of high hydrostatic pressure on rat atrial muscle.
Muscle preparations from rat atria were hydraulically compressed in circulating Tris-buffered solution kept at 37 degrees C. Spontaneously beating preparations decreased their beating frequency with 37.3 +/- 13.5 beats/min (22.3% +/- 6.7%, P less than 0.001) and increased their force with 2.3 +/- 1.1 mN (48.6% +/- 17.5%, P less than 0.001) when they were compressed to 10 MPa (100 atm). Decompression gave values not significantly different from precompression control values. The increase in force could in part be explained by the hyperbaric bradycardia and negative force-frequency relation. The remaining force increase seemed to be an effect of the increase in hydrostatic pressure. Changes in action potential duration and Ca2+ availability for the contractile machinery are discussed as possible mechanisms. Electrically driven preparations increased their contraction force at compression if the stimulus strength was at least 175% of the threshold. At lower stimulus levels just above threshold and at higher frequencies, inconsistent results were obtained at pressure.


Application of Four Genotyping Methods to Mycoplasma bovis Isolates Derived from Western Canadian Feedlot Cattle.
Mycoplasma bovis is a significant pathogen of feedlot cattle, responsible for chronic pneumonia and polyarthritis syndrome (CPPS). M. bovis isolates (n = 129) were used to compare four methods of phylogenetic analysis and to determine if the isolates' genotypes were associated with phenotypes. Metadata included the health status of the animal from which an isolate was derived (healthy, diseased, or dead), anatomical location (nasopharynx, lung, or joint), feedlot, and production year (2006 to 2018). Four in silico phylogenetic typing methods were used: multilocus sequence typing (MLST), core genome MLST (cgMLST), core genome single nucleotide variant (cgSNV) analysis, and whole-genome SNV (wgSNV) analysis. Using Simpson's diversity index (D) as a proxy for resolution, MLST had the lowest resolution (D = 0.932); cgSNV (D = 0.984) and cgMLST (D = 0.987) generated comparable results; and wgSNV (D = 1.000) provided the highest resolution. Visual inspection of the minimum spanning trees found that the memberships of the clonal complexes and clades had similar structural appearances. Although MLST had the lowest resolution, this methodology was intuitive and easy to apply, and the PubMLST database facilitates the comparison of sequence types across studies. The cg methods had higher resolution than MLST, and the graphical interface software was user-friendly for nonbioinformaticians, but the proprietary software is relatively expensive. The wgSNV approach was the most robust for processing poor-quality sequence data while offering the highest resolution; however, application of its software requires specialized training. None of the four methods could associate genotypes with phenotypes.


Theodore E. Woodward award. The evolution of obesity: insights from the mid-Miocene.
All humans are double knockouts. Humans lack the ability to synthesize vitamin C due to a mutation in L-gulono-lactone oxidase that occurred during the late Eocene, and humans have higher serum uric acid levels due to a mutation in uricase that occurred in the mid Miocene. In this paper we review the hypothesis that these mutations have in common the induction of oxidative stress that may have had prosurvival effects to enhance the effects of fructose to increase fat stores. Fructose was the primary nutrient in fruit which was the main staple of early primates, but this food likely became less available during the global cooling that occurred at the time of these mutations. However, in today's society, the intake of fructose, primarily in the form of added sugars, has skyrocketed, while the intake of natural fruits high in vitamin C has fallen. We suggest that it is the interaction of these genetic changes with diet that is responsible for the obesity epidemic today. Hence, we propose that Neel's thrifty gene hypothesis is supported by these new insights into the mechanisms regulating fructose metabolism.


Laser in situ keratomileusis for the correction of hyperopia from +0.50 to +11.50 diopters with the Keracor 117C laser.
To evaluate excimer laser in situ keratomileusis (LASIK) for the correction of hyperopia. We reviewed retrospectively the medical records of 46 patients treated with LASIK for hyperopia. All patients had a complete ophthalmologic evaluation. The corneal bed was ablated using the Bausch & Lomb Chiron Keracor 117C excimer laser to create a paracentral annular ablation under a nasally hinged 160-microm corneal flap with the Chiron Automatic Corneal Shaper microkeratome. Follow-up was a minimum of 6 months. Eighty eyes of 46 patients (23 males and 23 females) were included. Age ranged from 18 to 65 years (mean, 42 yr). The range of preoperative spherical equivalent refraction was +0.50 to +11.50 D (mean, +3.40 D). Mean postoperative spherical equivalent refraction at 6 months was +0.26 D. Six months after surgery, 35 eyes (44%) achieved uncorrected visual acuity of 20/20 or better and 78 eyes (97.5%) achieved 20/40 or better. Forty-six eyes (58%) had a postoperative spherical equivalent refraction within +/-0.50 D of attempted correction, and 67 eyes (84%) were within +/-1.00 D of attempted correction. When using the Bausch & Lomb Chiron Keracor 117C excimer laser to correct hyperopia, eyes with a spherical equivalent refraction less +2.00 D should be overcorrected by 25%, +2.00 to +4.00 D by 30%, and over +4.00 by 40%. The positive cylinder should be overcorrected by 10%. LASIK was safe and effective in the treatment of hyperopia from +0.50 to +11.50 D. Regression following LASIK for hyperopia remains a problem. A special nomogram was required to achieve results comparable with those for myopia.


A novel hydroxysuberamide derivative potentiates MG132-mediated anticancer activity against human hormone refractory prostate cancers--the role of histone deacetylase and endoplasmic reticulum stress.
Histone deacetylase (HDAC) inhibitors are successful for treatment of advanced cutaneous T-cell lymphoma but only show modest effect in solid tumors. Approaches for HDAC inhibitors to improve activity against solid tumors are necessary. Sulforhodamine B assay and flow cytometric analysis detected cell proliferation and cell-cycle progression, respectively. Protein expression was determined by Western blotting. Comet assay and DNA end-binding activity of Ku proteins detected DNA damage and DNA repair activity, respectively. siRNA technique was used for knockdown of specific cellular target. WJ25591 displayed inhibitory activity against HDAC1 and cell proliferation in human hormone-refractory prostate cancers PC-3 and DU-145. WJ25591 caused an arrest of cell-cycle at both G1- and G2-phase and increased protein expressions of p21 and cyclin E, followed by cell apoptosis. WJ25591-induced Bcl-2 down-regulation and activation of caspase-9, -8, and -3, suggesting apoptotic execution through both intrinsic and extrinsic apoptotic pathways. WJ25591 also significantly inhibited DNA repair activity but not directly induced DNA damage. Moreover, the proteasome inhibitor MG-132 dramatically sensitized WJ25591-induced cell apoptosis. The siRNA technique demonstrated that endoplasmic reticulum (ER) stress, in particular CHOP/GADD153 up-regulation, contributed to the synergistic effect. The data suggest that WJ25591 inhibited HDAC activity, leading to cell-cycle arrest and inhibition of DNA repair. Caspase cascades are subsequently triggered to execute cell apoptosis. MG-132 dramatically sensitizes WJ25591-mediated apoptosis, at least partly, through ER stress response. The data also reveal that combination of HDAC inhibitors and proteasome inhibitors may be a potential strategy against hormone-refractory prostate cancers.


Alterations in RFamide-related peptide expression are coordinated with the preovulatory luteinizing hormone surge.
The preovulatory LH surge is triggered when the circadian pacemaker, the bilateral suprachiasmatic nucleus (SCN), stimulates the GnRH system in the presence of high estrogen concentrations (positive feedback). Importantly, during the remainder of the estrous cycle, estradiol inhibits LH release via negative feedback. We have recently documented the presence of a novel mammalian RFamide-related peptide (RFRP), a putative gonadotropin-inhibitory hormone (GnIH), that presumably acts upstream of GnRH to modulate the negative feedback effects of estrogen. The present series of studies used female Syrian hamsters to examine the possibility that, in addition to driving the LH surge positively, the SCN concomitantly coordinates the removal of steroid-mediated RFRP inhibition of the gonadotropic axis to permit the surge. We found that the SCN forms close appositions with RFRP cells, suggesting the possibility for direct temporal control of RFRP activity. During the time of the LH surge, immediate-early gene expression is reduced in RFRP cells, and this temporal regulation is estrogen dependent. To determine whether projections from the SCN regulate the timed reduction in activation of the RFRP system, we exploited the phenomenon of splitting. In split animals in which the SCN are active in antiphase, activation of the RFRP system is asymmetrical. Importantly, this asymmetry is opposite to the state of the GnRH system. Together, these findings point to novel circadian control of the RFRP system and potential participation in the circuitry controlling ovulatory function.


Molecular Subtypes and the Evolution of Treatment Decisions in Metastatic Colorectal Cancer.
Colorectal cancer (CRC) has clinically relevant molecular heterogeneity at multiple levels: genomics, epigenomics, transcriptomics, and microenvironment features. Genomic events acquired during carcinogenesis remain drivers of cancer progression in the metastatic setting. For example, KRAS and NRAS mutations define a population refractory to epidermal growth factor receptor monoclonal antibodies, BRAFV600E mutations associate with poor outcomes under standard therapies and response to targeted inhibitors in combinations, and HER2 amplifications confer unique sensitivity to double HER2 blockade. Multiple rare gene alterations driving resistance to epidermal growth factor receptor monoclonal antibodies have been described, with substantial overlap in primary and acquired mechanisms, in line with a clonal selection process. In this context, sequential analysis of circulating tumor DNA has the potential to guide drug development in a treatment-refractory setting. Rare kinase fusion events and complex alterations in genes involved in DNA damage repair have been described, with emerging evidence for targetability. On the other hand, transcriptomic subtypes and pathway activation signatures have also shown prognostic and potential predictive value in metastatic CRC. These markers reflect stromal and immune microenvironment interactions with cancer cells. For example, the microsatellite instable or POLE ultramutant CRC population is particularly sensitive to immune checkpoint inhibitors, whereas tumors with a mesenchymal phenotype are characterized by activation of immunosuppressive molecules that mandate stratified development of novel immunotherapy combinations. Here, we review the expanding landscape of targetable oncogenic alterations and signatures in metastatic CRC and discuss the clinical implementation of novel molecular diagnostic tests.


Effective pulmonary delivery of an aerosolized plasmid DNA vaccine via surface acoustic wave nebulization.
Pulmonary-delivered gene therapy promises to mitigate vaccine safety issues and reduce the need for needles and skilled personnel to use them. While plasmid DNA (pDNA) offers a rapid route to vaccine production without side effects or reliance on cold chain storage, its delivery to the lung has proved challenging. Conventional methods, including jet and ultrasonic nebulizers, fail to deliver large biomolecules like pDNA intact due to the shear and cavitational stresses present during nebulization. In vitro structural analysis followed by in vivo protein expression studies served in assessing the integrity of the pDNA subjected to surface acoustic wave (SAW) nebulisation. In vivo immunization trials were then carried out in rats using SAW nebulized pDNA (influenza A, human hemagglutinin H1N1) condensate delivered via intratracheal instillation. Finally, in vivo pulmonary vaccinations using pDNA for influenza was nebulized and delivered via a respirator to sheep. The SAW nebulizer was effective at generating pDNA aerosols with sizes optimal for deep lung delivery. Successful gene expression was observed in mouse lung epithelial cells, when SAW-nebulized pDNA was delivered to male Swiss mice via intratracheal instillation. Effective systemic and mucosal antibody responses were found in rats via post-nebulized, condensed fluid instillation. Significantly, we demonstrated the suitability of the SAW nebulizer to administer unprotected pDNA encoding an influenza A virus surface glycoprotein to respirated sheep via aerosolized inhalation. Given the difficulty of inducing functional antibody responses for DNA vaccination in large animals, we report here the first instance of successful aerosolized inhalation delivery of a pDNA vaccine in a large animal model relevant to human lung development, structure, physiology, and disease, using a novel, low-power (<1 W) surface acoustic wave (SAW) hand-held nebulizer to produce droplets of pDNA with a size range suitable for delivery to the lower respiratory airways.


MicroRNAs as a Potential Quality Measurement Tool of Platelet Concentrate Stored in Blood Banks-A Review.
Platelet concentrate (PC) is one of the main products used in a therapeutic transfusion. This blood component requires special storage at blood banks, however, even under good storage conditions, modifications or degradations may occur and are known as platelet storage lesions. This research was performed on scientific citation databases PubMed/Medline, ScienceDirect, and Web of Science, for publications containing platelet storage lesions. The results obtained mainly reveal the clinical applicability of miRNAs as biomarkers of storage injury and as useful tools for a problem affecting public and private health, the lack of PC bags in countries with few blood donors. The major studies listed in this review identified miRNAs associated with important platelet functions that are relevant in clinical practice as quality biomarkers of PC, such as miR-223, miR-126, miR-10a, miR-150, miR-16, miR-21, miR-326, miR-495, let-7b, let-7c, let-7e, miR-107, miR-10b, miR-145, miR-155, miR-17, miR-191, miR-197, miR-200b, miR-24, miR-331, miR-376. These miRNAs can be used in blood banks to identify platelet injury in PC bags. The studies described in this review relate the functions of miRNAs with molecular mechanisms that result in functional platelet differences, such as apoptosis. Thus, miRNA profiles can be used to measure the quality of storage PC for more than 5 days, identify bags with platelet injury, and distinguish those with functional platelets.


Osmotic Stress-Induced Defective Glial Proteostasis Contributes to Brain Demyelination after Hyponatremia Treatment.
Adequate protein folding is necessary for normal cell function and a tightly regulated process that requires proper intracellular ionic strength. In many cell types, imbalance between protein synthesis and degradation can induce endoplasmic reticulum (ER) stress, which if sustained, can in turn lead to cell death. In nematodes, osmotic stress induces massive protein aggregation coupled with unfolded protein response and ER stress. In clinical practice, patients sustaining rapid correction of chronic hyponatremia are at risk of osmotic demyelination syndrome. The intense osmotic stress sustained by brain cells is believed to be the major risk factor for demyelination resulting from astrocyte death, which leads to microglial activation, blood-brain barrier opening, and later, myelin damage. Here, using a rat model of osmotic demyelination, we showed that rapid correction of chronic hyponatremia induces severe alterations in proteostasis characterized by diffuse protein aggregation and ubiquitination. Abrupt correction of hyponatremia resulted in vigorous activation of both the unfolded protein response and ER stress accompanied by increased autophagic activity and apoptosis. Immunofluorescence revealed that most of these processes occurred in astrocytes within regions previously shown to be demyelinated in later stages of this syndrome. These results identify osmotic stress as a potent protein aggregation stimuli in mammalian brain and further suggest that osmotic demyelination might be a consequence of proteostasis failure on severe osmotic stress.


The 1998 Lindberg Award. Comparison of glycerol preservation with cryopreservation methods on HIV-1 inactivation.
Cryopreservation and glycerol preservation are 2 successful methods for long-term preservation of human cadaver skin. Preservation is subjected to strict criteria to minimize the risk of disease transmission. This investigation compares the effects of glycerol preservation and cryopreservation on the inactivation of HIV-1. The effects of glycerol preservation and cryopreservation on inactivation of both extracellular and intracellular HIV-1Ba-L were investigated. After exposing HIV-1Ba-L-infected material to various concentrations of glycerol or to 10% dimethyl sulfoxide followed by cryopreservation, uninfected peripheral blood mononuclear cells were added to the treated material. At different time points during the culture, supernatants were taken to quantify HIV-1Ba-L and reverse transcriptase levels to determine HIV-1Ba-L infectivity. Cell-free HIV-1Ba-L was inactivated within 30 minutes in 70% and 85% glycerol. Also, intracellular HIV-1Ba-L in infected peripheral blood mononuclear cells or infected cadaver skin was completely inactivated by glycerol treatment in vitro. Cryopreservation did not show any extracellular or intracellular HIV-1Ba-L inactivation. Glycerol preservation--but not cryopreservation--of human cadaveric donor skin can inactivate both extracellular and intracellular HIV-1.


Predictive validity and efficiency of ongoing visual-inspection criteria for interpreting functional analyses.
Prior research has evaluated the reliability and validity of structured criteria for visually inspecting functional-analysis (FA) results on a post-hoc basis, after completion of the FA (i.e., post-hoc visual inspection [PHVI]; e.g., Hagopian et al., 1997). However, most behavior analysts inspect FAs using ongoing visual inspection (OVI) as the FA is implemented, and the validity of applying structured criteria during OVI remains unknown. In this investigation, we evaluated the predictive validity and efficiency of applying structured criteria on an ongoing basis by comparing the interim interpretations produced through OVI with (a) the final interpretations produced by PHVI, (b) the authors' post-hoc interpretations (PHAI) reported in the research studies, and (c) the consensus interpretations of these two post-hoc analyses. Ongoing visual inspection predicted the results of PHVI and the consensus interpretations with a very high degree of accuracy, and PHAI with a reasonably high degree of accuracy. Furthermore, the PHVI and PHAI results involved 32 FA sessions, on average, whereas the OVI required only 19 FA sessions to accurately identify the function(s) of destructive behavior (i.e., a 41% increase in efficiency). We discuss these findings relative to other methods designed to increase the accuracy and efficiency of FAs.


Prion protein self-peptides modulate prion interactions and conversion.
Molecular mechanisms underlying prion agent replication, converting host-encoded cellular prion protein (PrP(C)) into the scrapie associated isoform (PrP(Sc)), are poorly understood. Selective self-interaction between PrP molecules forms a basis underlying the observed differences of the PrP(C) into PrP(Sc) conversion process (agent replication). The importance of previously peptide-scanning mapped ovine PrP self-interaction domains on this conversion was investigated by studying the ability of six of these ovine PrP based peptides to modulate two processes; PrP self-interaction and conversion. Three peptides (octarepeat, binding domain 2 -and C-terminal) were capable of inhibiting self-interaction of PrP in a solid-phase PrP peptide array. Three peptides (N-terminal, binding domain 2, and amyloidogenic motif) modulated prion conversion when added before or after initiation of the prion protein misfolding cyclic amplification (PMCA) reaction using brain homogenates. The C-terminal peptides (core region and C-terminal) only affected conversion (increased PrP(res) formation) when added before mixing PrP(C) and PrP(Sc), whereas the octarepeat peptide only affected conversion when added after this mixing. This study identified the putative PrP core binding domain that facilitates the PrP(C)-PrP(Sc) interaction (not conversion), corroborating evidence that the region of PrP containing this domain is important in the species-barrier and/or scrapie susceptibility. The octarepeats can be involved in PrP(C)-PrP(Sc) stabilization, whereas the N-terminal glycosaminoglycan binding motif and the amyloidogenic motif indirectly affected conversion. Binding domain 2 and the C-terminal domain are directly implicated in PrP(C) self-interaction during the conversion process and may prove to be prime targets in new therapeutic strategy development, potentially retaining PrP(C) function. These results emphasize the importance of probable PrP(C)-PrP(C) and required PrP(C)-PrP(Sc) interactions during PrP conversion. All interactions are probably part of the complex process in which polymorphisms and species barriers affect TSE transmission and susceptibility.


Feigning combat-related posttraumatic stress disorder on the personality assessment inventory.
This study examined whether individuals who were instructed on the Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994) criteria for posttraumatic stress disorder (PTSD) could feign PTSD on the Personality Assessment Inventory (PAI; Morey, 1991). The study also investigated whether PAI indexes of symptom exaggeration, the Negative Impression Management (NIM) scale and the Malingering index, could identify individuals feigning PTSD. The diagnostic rule for PTSD (Morey, 1991, 1996) was applied to the profiles of a group of 23 veterans with combat-related PTSD and 23 male undergraduates instructed to malinger PTSD. Seventy percent of the student malingerers produced profiles that received diagnostic consideration for PTSD. The NIM cutting score (> or = 8) was highly effective in detecting simulation of PTSD but resulted in the misclassification of a large number of true PTSD cases. There were no significant differences in the overall efficiency of the test with various validity criteria. We discuss the implications of these findings for the use of the PAI in the diagnosis of combat-related PTSD.


Implant-retained precision two-stage single-tooth replacement.
The restoration of an anterior edentulous space due to a congenitally missing lateral incisor presents the dentist with a restorative challenge. Treatment of a young patient requires that the restoration be conservative as well as esthetic. This article presents the rationale for use of an osseo-integrated implant to support a single-tooth restoration. The final result was esthetic, functional, and promoted a healthy periodontium.


Genome-wide and parental allele-specific analysis of CTCF and cohesin DNA binding in mouse brain reveals a tissue-specific binding pattern and an association with imprinted differentially methylated regions.
DNA binding factors are essential for regulating gene expression. CTCF and cohesin are DNA binding factors with central roles in chromatin organization and gene expression. We determined the sites of CTCF and cohesin binding to DNA in mouse brain, genome wide and in an allele-specific manner with high read-depth ChIP-seq. By comparing our results with existing data for mouse liver and embryonic stem (ES) cells, we investigated the tissue specificity of CTCF binding sites. ES cells have fewer unique CTCF binding sites occupied than liver and brain, consistent with a ground-state pattern of CTCF binding that is elaborated during differentiation. CTCF binding sites without the canonical consensus motif were highly tissue specific. In brain, a third of CTCF and cohesin binding sites coincide, consistent with the potential for many interactions between cohesin and CTCF but also many instances of independent action. In the context of genomic imprinting, CTCF and/or cohesin bind to a majority but not all differentially methylated regions, with preferential binding to the unmethylated parental allele. Whether the parental allele-specific methylation was established in the parental germlines or post-fertilization in the embryo is not a determinant in CTCF or cohesin binding. These findings link CTCF and cohesin with the control regions of a subset of imprinted genes, supporting the notion that imprinting control is mechanistically diverse.


Continuous-Flow Laboratory SAXS for In Situ Determination of the Impact of Hydrophilic Block Length on Spherical Nano-Object Formation during Polymerization-Induced Self-Assembly.
In situ small-angle X-ray scattering (SAXS) is a powerful technique for characterizing block-copolymer nano-object formation during polymerization-induced self-assembly. To work effectively in situ, it requires high intensity X-rays which enable the short acquisition times required for real-time measurements. However, routine access to synchrotron X-ray sources is expensive and highly competitive. Flow reactors provide an opportunity to obtain temporal resolution by operating at a consistent flow rate. Here, we equip a flow-reactor with an X-ray transparent flow-cell at the outlet which facilitates the use of a low-flux laboratory SAXS instrument for in situ monitoring. The formation and morphological evolution of spherical block copolymer nano-objects was characterized during reversible addition fragmentation chain transfer polymerization of diacetone acrylamide in the presence of a series of poly(dimethylacrylamide) (PDMAm) macromolecular chain transfer agents with varying degrees of polymerization. SAXS analysis indicated that during the polymerization, highly solvated, loosely defined aggregates form after approximately 100 s, followed by expulsion of solvent to form well-defined spherical particles with PDAAm cores and PDMAm stabilizer chains, which then grow as the polymerization proceeds. Analysis also indicates that the aggregation number (Nagg) increases during the reaction, likely due to collisions between swollen, growing nanoparticles. In situ SAXS conducted on PISA syntheses using different PDMAm DPs indicated a varying conformation of the chains in the particle cores, from collapsed chains for PDMAm47 to extended chains for PDMAm143. At high conversion, the final Nagg decreased as a function of increasing PDMAm DP, indicating increased steric stabilization afforded by the longer chains which is reflected by a decrease in both core diameter (from SAXS) and hydrodynamic diameter (from DLS) for a constant core DP of 400.


MiR-181 family: regulators of myeloid differentiation and acute myeloid leukemia as well as potential therapeutic targets.
MicroRNAs have been shown to play an important role in normal hematopoisis and leukemogenesis. Here, we report function and mechanisms of miR-181 family in myeloid differentiation and acute myeloid leukemia (AML). The aberrant overexpression of all the miR-181 family members (miR-181a/b/c/d) was detected in French-American-British M1, M2 and M3 subtypes of adult AML patients. By conducting gain- and loss-of-function experiments, we demonstrated that miR-181a inhibits granulocytic and macrophage-like differentiation of HL-60 cells and CD34+ hematopoietic stem/progenitor cells (HSPCs) by directly targeting and downregulating the expression of PRKCD (which then affected the PRKCD-P38-C/EBPα pathway), CTDSPL (which then affected the phosphorylation of retinoblastoma protein) and CAMKK1. The three genes were also demonstrated to be the targets of miR-181b, miR-181c and miR-181d, respectively. Significantly decreases in the expression levels of the target proteins were detected in AML patients. Inhibition of the expression of miR-181 family members owing to Lenti-miRZip-181a infection in bone marrow blasts of AML patients increased target protein expression levels and partially reversed myeloid differentiation blockage. In the mice implanted with AML CD34+ HSPCs, expression inhibition of the miR-181 family by Lenti-miRZip-181a injection improved myeloid differentiation, inhibited engraftment and infiltration of the leukemic CD34+ cells into the bone marrow and spleen, and released leukemic symptoms. In conclusion, our findings revealed new mechanism of miR-181 family in normal hematopoiesis and AML development, and suggested that expression inhibition of the miR-181 family could provide a new strategy for AML therapy.


Isoform-Specific Phosphorylation in Human Hsp90β Affects Interaction with Clients and the Cochaperone Cdc37.
The 90-kDa heat shock proteins (Hsp90s) assist the maturation of many key regulators of signal transduction pathways and cellular control circuits like protein kinases and transcription factors and chaperone their stability and activity. In this function, Hsp90s cooperate with some 30 cochaperones and they are themselves subject to regulation by numerous post-translational modifications. In vertebrates, two major isoforms exist in the cytosol, Hsp90α and Hsp90β, which share a high degree of sequence identity and are expressed in tissue- and environmental condition-dependent manner. We identified an isoform-specific phosphorylation site in human Hsp90β. This phosphorylation site seems to be linked to vertebrate evolution since it is not found in invertebrata but in all tetrapoda and many but not all fish species. We provide data suggesting that this phosphorylation is important for the activation of Hsp90 clients like glucocorticoid receptor and a protein kinase. Replacement of the phosphorylation site by glutamate affects the conformational dynamics of Hsp90 and interaction with the kinase-specific cochaperone Cdc37.


Cretaceous Mammals from Montana.
A series of Cretaceous mammal faunas, beginning with standard late Cretaceous faunas and continuing with three of Paleocene aspect, are summarized. Four families (Eucosmodontidae, Taeniolabididae, Leptictidae, Arctocyonidae) and one order (Condylarthra) of Tertiary mammals are extended into the Cretaceous. New genera Cimexomys, Stygimys, Procerberus, and Protungulatum are described, as is a small species of Catopsalis. The skeleton of a multituberculate is restored and multituberculate classification is revised.


In vitro biocompatibility of dextrin: the addition of a low concentration of dextrin in the medium promotes the cell activity of L929 mouse fibroblasts.
To develop a bone substitute with shape-generating properties, we focused our attention on dextrin, which has a low viscosity. After considering methods of evaluation for research and development, we started by using cells that are widely used for safe biological evaluations in the field of dentistry and conducted in vitro evaluations. In this experiment, we variously added concentrations of 0.1, 1.0 and 10 mmol/l of dextrin to a culture medium in order to examine the effects on L929 mouse fibroblasts in vitro. As a result, the proliferative activity of the L929 cells was promoted during the culture period as the concentration of added dextrin became lower, and in particular, the 0.1 and 1 mmol/l addition group showed higher values than those of the control group. From the above results, it was revealed that the addition of a low concentration of dextrin in a medium promotes the cell proliferative activity.


Mapping whole genome shotgun sequence and variant calling in mammalian species without their reference genomes.
Genomics research in mammals has produced reference genome sequences that are essential for identifying variation associated with disease. High quality reference genome sequences are now available for humans, model species, and economically important agricultural animals. Comparisons between these species have provided unique insights into mammalian gene function. However, the number of species with reference genomes is small compared to those needed for studying molecular evolutionary relationships in the tree of life. For example, among the even-toed ungulates there are approximately 300 species whose phylogenetic relationships have been calculated in the 10k trees project. Only six of these have reference genomes: cattle, swine, sheep, goat, water buffalo, and bison. Although reference sequences will eventually be developed for additional hoof stock, the resources in terms of time, money, infrastructure and expertise required to develop a quality reference genome may be unattainable for most species for at least another decade. In this work we mapped 35 Gb of next generation sequence data of a Katahdin sheep to its own species' reference genome ( Ovis aries Oar3.1) and to that of a species that diverged 15 to 30 million years ago ( Bos taurus UMD3.1). In total, 56% of reads covered 76% of UMD3.1 to an average depth of 6.8 reads per site, 83 million variants were identified, of which 78 million were homozygous and likely represent interspecies nucleotide differences. Excluding genome repeat regions and sex chromosomes, approximately 3.7 million heterozygous sites were identified in this animal vs. bovine UMD3.1, representing polymorphisms occurring in sheep. Of these, 41% could be readily mapped to orthologous positions in ovine Oar3.1 with 80% corroborated as heterozygous. These variant sites, identified via interspecies mapping could be used for comparative genomics, disease association studies, and ultimately to understand mammalian gene function.


Drug resistance among newly diagnosed HIV-infected children in the era of more efficacious antiretroviral prophylaxis.
In the era of more efficacious prevention of mother-to-child transmission (PMTCT) regimens, documenting the profile of drug resistance in HIV-infected infants and young children is critical to our efforts to improve care and treatment for children. HIV drug resistance mutations in plasma virus were ascertained using population sequencing among 230 newly diagnosed HIV-infected children under 2 years of age recruited in Johannesburg, South Africa, during 2011. By this time, more effective PMTCT regimens, including combination antiretroviral therapy for pregnant women, were being implemented. Two-thirds (67.4%) of HIV-infected children had been exposed to some form of maternal (89%) and/or infant (97%) PMTCT. Among PMTCT-exposed, 56.8% had nonnucleoside reverse transcriptase inhibitor (NNRTI), 14.8% nucleoside reverse transcriptase inhibitor (NRTI), and 1.3% protease inhibitor mutations. NNRTI mutations were strongly related to younger age. The remaining third (32.6%) had no reported or recorded PMTCT exposures, but resistance to NNRTI was detected in 24.0%, NRTI in 10.7%, and protease inhibitor in 1.3%. The new PMTCT strategies dramatically reduce the number of children who acquire infection, but among those who do become infected, NNRTI resistance prevalence is high. In this South African setting with high PMTCT coverage, almost a quarter of children with no reported or recorded PMTCT also have drug resistance mutations. PMTCT history is an inadequate means of ruling out pretreatment drug resistance. Our results support the use of protease inhibitor-based first-line regimens in HIV-infected infants and young children regardless of PMTCT history.


Immunoendocrinology: faulty hormonal imprinting in the immune system.
Hormonal imprinting is an epigenetic process which is taking place perinatally at the first encounter between the developing hormone receptors and their target hormones. The hormonal imprinting influences the binding capacity of receptors, the hormone synthesis of the cells, and other hormonally regulated functions, as sexual behavior, aggressivity, empathy, etc. However, during the critical period, when the window for imprinting is open, molecules similar to the physiological imprinters as synthetic hormone analogs, other members of the hormone families, environmental pollutants, etc. can cause faulty imprinting with life-long consequences. The developing immune system, the cells of which also have receptors for hormones, is very sensitive to faulty imprinting, which causes alterations in the antibody and cytokine production, in the ratio of immune cells, in the defense against bacterial and viral infections as well as against malignant tumors. Immune cells (lymphocytes, monocytes, granulocytes and mast cells) are also producing hormones which are secreted into the blood circulation as well as are transported locally (packed transport). This process is also disturbed by faulty imprinting. As immune cells are differentiating during the whole life, faulty imprinting could develop any time, however, the most decisive is the perinatal imprinting. The faulty imprinting is inherited to the progenies in general and especially in the case of immune system. In our modern world the number and amount of artificial imprinters (e.g. endocrine disruptors and drugs) are enormously increasing. The effects of the faulty imprinters most dangerous to the immune system are shown in the paper. The present and future consequences of the flood of faulty imprintings are unpredictable however, it is discussed.


Opinions of trauma practitioners regarding prehospital interventions for critically injured patients.
Significant controversy surrounds the prehospital management of trauma patients. A questionnaire describing clinical scenarios was mailed to a random sample of 345 trauma practitioners. The 182 trauma practitioners (52.8%) who returned the surveys were predominantly general or trauma surgeons (83.5%) in academic or university practice (68.1%). For a patient with a severe traumatic brain injury, 84.5% of trauma practitioners recommended that emergency medical services personnel attempt intubation at least once when transport time was 20 to 40 minutes. For a patient with a gunshot wound to the epigastrium in decompensated shock, the majority of trauma practitioners believed that a relatively hypotensive state should be maintained, regardless of transport time. Trauma practitioners (52.2%) have recommended the use of the pneumatic antishock garment for transports of 20 to 40 minutes for patients with an unstable pelvic fracture and decompensated shock. Most trauma practitioners believe that emergency medical services providers should attempt intubation for a patient with a severe traumatic brain injury, should treat decompensated shock in a patient with penetrating torso trauma but maintain the patient in a relatively hypotensive state, and should apply and inflate the pneumatic antishock garment for a suspected pelvic fracture accompanied by decompensated shock if the patient is 20 to 40 minutes from a trauma center. The recommendations of trauma practitioners regarding appropriate prehospital care are significantly influenced by the time required for transport to the trauma center.


Auditory and Audiovisual Close Shadowing in Post-Lingually Deaf Cochlear-Implanted Patients and Normal-Hearing Elderly Adults.
The goal of this study was to determine the effect of auditory deprivation and age-related speech decline on perceptuo-motor abilities during speech processing in post-lingually deaf cochlear-implanted participants and in normal-hearing elderly (NHE) participants. A close-shadowing experiment was carried out on 10 cochlear-implanted patients and on 10 NHE participants, with two groups of normal-hearing young participants as controls. To this end, participants had to categorize auditory and audiovisual syllables as quickly as possible, either manually or orally. Reaction times and percentages of correct responses were compared depending on response modes, stimulus modalities, and syllables. Responses of cochlear-implanted subjects were globally slower and less accurate than those of both young and elderly normal-hearing people. Adding the visual modality was found to enhance performance for cochlear-implanted patients, whereas no significant effect was obtained for the NHE group. Critically, oral responses were faster than manual ones for all groups. In addition, for NHE participants, manual responses were more accurate than oral responses, as was the case for normal-hearing young participants when presented with noisy speech stimuli. Faster reaction times were observed for oral than for manual responses in all groups, suggesting that perceptuo-motor relationships were somewhat successfully functional after cochlear implantation and remain efficient in the NHE group. These results are in agreement with recent perceptuo-motor theories of speech perception. They are also supported by the theoretical assumption that implicit motor knowledge and motor representations partly constrain auditory speech processing. In this framework, oral responses would have been generated at an earlier stage of a sensorimotor loop, whereas manual responses would appear late, leading to slower but more accurate responses. The difference between oral and manual responses suggests that the perceptuo-motor loop is still effective for NHE subjects and also for cochlear-implanted participants, despite degraded global performance.


Current Frontline Treatment of Diffuse Large B-Cell Lymphoma.
Diffuse large B-cell lymphoma (DLBCL), the most common subtype of non-Hodgkin lymphoma, is an aggressive and biologically heterogeneous disease. Risk stratification and treatment algorithms vary based on stage of disease and bulk along with other clinical and biological factors, including the International Prognostic Index, cell of origin, and other molecular subsets. Rituximab (Rituxan), cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) is the current standard of care and cures more than 60% of patients. The role of radiotherapy is largely restricted to patients with limited-stage disease. In elderly patients, geriatric assessments of baseline fitness and functional status help optimize therapy based on the balance of efficacy and toxicity. While numerous randomized trials have failed to improve upon R-CHOP, a recent press release from the POLARIX trial (NCT03274492) suggests that adding polatuzumab vedotin (Polivy) to rituximab, cyclophosphamide, doxorubicin, and prednisone (pola-R-CHP) improves progression-free survival and may replace R-CHOP in eligible patients. Ongoing trials are exploring frontline therapy that integrates other novel agents, including various small molecules, bispecific antibodies, and chimeric antigen receptor T-cell therapy, with promising preliminary results. Defining a population of patients with high-risk disease in whom R-CHOP is not effective is critical. Patient selection based on refining molecular subsets, quantitative PET metrics such as metabolic tumor volume, and dynamic risk assessments using interim PET and circulating tumor DNA analysis may allow for a personalized, response-adapted approach that will further improve outcomes in DLBCL.


An inexpensive and worldwide available digital image analysis technique for histological fibrosis quantification in chronic hepatitis C.
Hepatic fibrosis staging is based on semiquantitative scores. Digital imaging analysis (DIA) appears more accurate because fibrosis is quantified in a continuous scale. However, high cost, lack of standardization and worldwide unavailability restrict its use in clinical practice. We developed an inexpensive and widely available DIA technique for fibrosis quantification in hepatitis C, and here, we evaluate its reproducibility and correlation with semiquantitative scores, and determine the fibrosis percentage associated with septal fibrosis and cirrhosis. 282 needle biopsies staged by Ishak and METAVIR scores were included. Images of trichrome-stained sections were captured and processed using Adobe(®) Photoshop(®) CS3 and Adobe(®) Bridge(®) softwares. The percentage of fibrosis (fibrosis index) was determined by the ratio between the fibrosis area and the total sample area, expressed in pixels calculated in an automated way. An excellent correlation between DIA fibrosis index and Ishak and METAVIR scores was observed (Spearman's r = 0.95 and 0.92; P < 0.001, respectively). Excellent intra-observer reproducibility was observed in a randomly chosen subset of 39 biopsies with an intraclass correlation index of 0.99 (95% CI, 0.95-0.99). The best cut-offs associated with septal fibrosis and cirrhosis were 6% (AUROC 0.97, 95% CI, 0.95-0.99) and 27% (AUROC 1.0, 95% CI, 0.99-1), respectively. This new DIA technique had high correlation with semiquantitative scores in hepatitis C. This method is reproducible, inexpensive and available worldwide allowing its use in clinical practice. The incorporation of DIA technique provides a more complete evaluation of fibrosis adding the quantification to architectural patterns.


Nanoscale structural and mechanical properties of nontypeable Haemophilus influenzae biofilms.
Nontypeable Haemophilus influenzae (NTHI) bacteria are commensals in the human nasopharynx, as well as pathogens associated with a spectrum of acute and chronic infections. Two important factors that influence NTHI pathogenicity are their ability to adhere to human tissue and their ability to form biofilms. Extracellular polymeric substances (EPS) and bacterial appendages such as pili critically influence cell adhesion and intercellular cohesion during biofilm formation. Structural components in the outer cell membrane, such as lipopolysaccharides, also play a fundamental role in infection of the host organism. In spite of their importance, these pathogenic factors are not yet well characterized at the nanoscale. Here, atomic force microscopy (AFM) was used in aqueous environments to visualize structural details, including probable Hif-type pili, of live NTHI bacteria at the early stages of biofilm formation. Using single-molecule AFM-based spectroscopy, the molecular elasticities of lipooligosaccharides present on NTHI cell surfaces were analyzed and compared between two strains (PittEE and PittGG) with very different pathogenicity profiles. Furthermore, the stiffness of single cells of both strains was measured and subsequently their turgor pressure was estimated.


Vitamin D3 transactivates the zinc and manganese transporter SLC30A10 via the Vitamin D receptor.
Vitamin D3 regulates genes critical for human health and its deficiency is associated with an increased risk for osteoporosis, cancer, diabetes, multiple sclerosis, hypertension, inflammatory and immunological diseases. To study the impact of vitamin D3 on genes relevant for the transport and metabolism of nutrients and drugs, we employed next-generation sequencing (NGS) and analyzed global gene expression of the human-derived Caco-2 cell line treated with 500nM vitamin D3. Genes involved in neuropeptide signaling, inflammation, cell adhesion and morphogenesis were differentially expressed. Notably, genes implicated in zinc, manganese and iron homeostasis were largely increased by vitamin D3 treatment. An ∼10-fold increase in ceruloplasmin and ∼4-fold increase in haptoglobin gene expression suggested a possible association between vitamin D and iron homeostasis. SLC30A10, the gene encoding the zinc and manganese transporter ZnT10, was the chiefly affected transporter, with ∼15-fold increase in expression. SLC30A10 is critical for zinc and manganese homeostasis and mutations in this gene, resulting in impaired ZnT10 function or expression, cause manganese intoxication, with Parkinson-like symptoms. Our NGS results were validated by real-time PCR in Caco-2 cells, as well as in duodenal biopsies taken from healthy human subjects treated with 0.5μg vitamin D3 daily for 10 days. In addition to increasing gene expression of SLC30A10 and the positive control TRPV6, vitamin D3 also increased ZnT10 protein expression, as indicated by Western blot and cytofluorescence. In silico identification of potential vitamin D responsive elements (VDREs) in the 5'-flanking region of the SLC30A10 promoter and dual-luciferase reporter assay showed enhanced promoter activity in the presence of vitamin D receptor (VDR) and retinoid X receptor (RXR) constructs, as well as vitamin D3, but not when one of these factors was absent. Electrophoretic mobility shift assay (EMSA) and competition EMSA revealed binding of select sequences, namely, nt -1623/-1588 and nt -1758/-1723 relative to the transcription start site, to VDR-containing nuclear extracts. In conclusion, we have shown that vitamin D3 transactivates the SLC30A10 gene in a VDR-dependent manner, resulting in increased ZnT10 protein expression. Because SLC30A10 is highly expressed in the small intestine, it is possible that the control of zinc and manganese systemic levels is regulated by vitamin D3 in the intestine. Zinc, manganese and vitamin D are important for bone metabolism and brain health. Future examination of a possible role for supplementation or chelation of zinc and manganese, alongside vitamin D3 administration, will further our understanding of its potential benefit in the treatment of specific illnesses, such as osteoporosis and Parkinson's disease.


A2Hgx(SeO3)y (A = K, Rb, Cs): Three Alkali Metal Mercury Selenites Featuring Unique 1D [HgOm(SeO3)n]∞ Chains.
Herein, three alkali metal mercury selenites, K2Hg2(SeO3)3, Rb2Hg2(SeO3)3, and Cs2Hg3(SeO3)4, were successfully obtained by a hydrothermal method. The three compounds featured same one-dimensional (1D) [HgOm(SeO3)n]∞ chain structure that consisting of distorted Hg-O polyhedra and SeO3 triangular pyramids with stereochemically active lone pair (SCALP) electrons. Interestingly, the rich coordination environment of Hg atoms and the size difference of alkali metal cations lead to diverse arrangement of SeO3 groups, which makes them exhibit different birefringence. The band gaps of the three compounds indicate that they are potential ultraviolet (UV) optical materials. Detailed theoretical calculations demonstrate that the combined effects of SeO3 triangular pyramids and Hg-O polyhedra are responsible for the optical characteristics of the reported compounds.


Characterization and application of calcium-dependent β-propeller phytase from Bacillus amyloliquefaciens DS11.
The enzyme phytase has broad biotechnological applications, especially in the reduction of phytate, antinutritional factors that chelate essential minerals, in human and animal food. We investigated the enzymatic properties of β-propeller phytase (BPP) from Bacillus amyloliquefaciens DS11. Thermal refolding analysis demonstrated that BPP can remarkably restore its enzymatic activity in the presence of 5 mM Ca(2+) to 87% of its original activity after heating to 100 °C and subsequent cooling, indicating that the enzyme requires Ca(2+) for appropriate refolding. Furthermore, pH-dependent kinetic studies showed that BPP required excess Ca(2+) for its enzymatic activity as the pH decreased, suggesting that the optimal Ca(2+)-phytate ratio for enzymatic catalysis depends on the pH value of the environment. Finally, we verified the practical application of BPP at two different pH's using soybean meal as a natural source of phytate. As compared to a commercial phytase, BPP efficiently hydrolyzed food phytate over neutral pH ranges.


Central nervous system endothelial cell-polymorphonuclear cell interactions during autoimmune demyelination.
The homing and adhesion of circulating cells to target tissue vasculature precedes their subsequent invasion of inflamed tissue. Polymorphonuclear cells (PMNs), key players in most inflammatory events, are among the first cells to arrive. The present work, performed on CNS lesions from mice with experimental autoimmune encephalomyelitis, provides morphologic evidence for interactions between PMNs and unique, frondlike extensions from endothelial cells (EC) during early attachment. Platelets also were seen attached to these endothelial fronds. The structures projected into vessel lumina from the vicinity of tight junctions and were often branched and complex, the latter characteristics suggesting a possible role in cellular 'trapping'. Polymorphonuclear cells appeared to traverse the CNS vasculature between EC where the blood-brain barrier was severely compromised with junctional complexes reduced to simple contact points. The cells from which the fronds derived were often plump and possessed cytoplasm rich in organelles, perhaps indicative of activation. The present report contrasts with previous observations on lymphocytes in the same system where lymphocytic pseudopodia formed intimate contacts before their burrowing directly through the endothelium and where EC fronds were not involved.


Palmatine ameliorated murine colitis by suppressing tryptophan metabolism and regulating gut microbiota.
Inflammatory bowel disease (IBD), majorly include Crohn's disease (CD) and ulcerative colitis (UC), is chronic and relapsing inflammatory disorders of the gastrointestinal tract, which treatment options remain limited. Here we examined the therapeutic effects of an isoquinoline alkaloid, Palmatine (Pal), on mice experimental colitis induced by dextran sulfate sodium (DSS) and explored underlying mechanisms. Colitis was induced in BALB/c mice by administering 3% DSS in drinking water for 7 days. Pal (50 and 100 mg kg-1) and the positive drug Sulfasalazine (SASP, 200 mg kg-1) were orally administered for 7 days. Disease activity index (DAI) was evaluated on day 8, and colonic tissues were collected for biochemistry analysis. The fecal microbiota was characterized by high-throughput Illumina MiSeq sequencing. And plasma metabolic changes were detected by UPLC-MS. Our results showed that Pal treatment significantly reduced DAI scores and ameliorated colonic injury in mice with DSS-induced colitis. Mucosal integrity was improved and cell apoptosis was inhibited. Moreover, gut microbiota analysis showed that mice received Pal-treatment have higher relative abundance of Bacteroidetes and Firmicutes, but reduced amount of Proteobacteria. Moreover, Pal not only suppressed tryptophan catabolism in plasma, but also decreased the protein expression of indoleamine 2,3-dioxygenase 1 (IDO-1, the rate-limiting enzyme of tryptophan catabolism) in colon tissue. This is consolidated by molecular docking, which suggested that Pal is a potent IDO-1 inhibitor. Taken together, our findings demonstrate that Pal ameliorated DSS-induced colitis by mitigating colonic injury, preventing gut microbiota dysbiosis, and regulating tryptophan catabolism, which indicated that Pal has great therapeutic potential for colitis.


Toward a leukemia treatment strategy based on the probability of stem cell death: an essay in honor of Dr. Emil J Freireich.
Dr. Emil J Freireich is a pioneer in the rational treatment of cancer in general and leukemia in particular. This essay in his honor suggests that the cell kill concept of chemotherapy of acute myeloblastic leukemia be extended to include two additional ideas. The first concept is that leukemic blasts, like normal hemopoietic cells, are organized in hierarchies, headed by stem cells. In both normal and leukemic hemopoiesis, killing stem cells will destroy the system; furthermore, both normal and leukemic cells respond to regulators. It follows that acute myelogenous leukemia should be considered as a dependent neoplasm. The second concept is that cell/drug interaction should be considered as two phases. The first, or proximal phase, consists of the events that lead up to injury; the second, or distal phase, comprises the responses of the cell that contribute to either progression to apoptosis or recovery. Distal responses are described briefly. Regulated drug sensitivity is presented as an example of how distal responses might be used to improve treatment.


Monitoring the surface tension by the pendant drop technique for detection of insulin fibrillogenesis.
Monitoring the aggregation of amyloid-prone proteins is critical for understanding the mechanism of amyloid fibril formation. Insulin, when dissolved in low pH buffer, has a surface tension of 61-64 mN m-1, as measured by the pendant drop technique. Formation of insulin amyloid fibrils resulted in the increase of the surface tension values up to 71.2-73.5 mN m-1. The kinetics of fibril formation and fibril morphology were validated by ThT fluorescence and AFM, respectively. The results demonstrate that monitoring the surface tension by the pendant drop technique is a valuable tool for the detection of insulin amyloid aggregation.


Correlation between agility and sprinting according to student age.
The purposes of the study were to assess sprinting and agility performance characteristics and to determine the relationship between these two motor skills in elementary education students. Sprinting and agility performance were assessed in 176 children (88 boys and 88 girls) divided into three groups: Group 1 (G1, N = 98; 48 boys and 50 girls), from the first year of elementary education; Group 2 (G2, N = 38; 15 boys and 23 girls), from the second year of elementary education; Group 3 (G3, N = 40; 25 boys and 15 girls), from the third year of elementary education. Significant differences (p < 0.001) were found in agility ability among the groups and between G1-G3 and G2-G3 in the 5 and 15 m sprint. Regarding gender of the students of the same age, significant differences (p < 0.001) between boys and girls in group G1 and G2 were obtained in the 5 and 15 m sprint. The correlation between agility and acceleration was significant but moderate (0.3 < r < 0.7) in all groups (G1, G2, and G3), in most cases. When the gender factor was included, the results were heterogeneous. Assessing this correlation according to age and gender produced heterogeneous results. For this reason, we think that both are independent qualities and that age and gender are two factors that influence the correlation results.


Evolution of genome size and complexity in the rhabdoviridae.
RNA viruses exhibit substantial structural, ecological and genomic diversity. However, genome size in RNA viruses is likely limited by a high mutation rate, resulting in the evolution of various mechanisms to increase complexity while minimising genome expansion. Here we conduct a large-scale analysis of the genome sequences of 99 animal rhabdoviruses, including 45 genomes which we determined de novo, to identify patterns of genome expansion and the evolution of genome complexity. All but seven of the rhabdoviruses clustered into 17 well-supported monophyletic groups, of which eight corresponded to established genera, seven were assigned as new genera, and two were taxonomically ambiguous. We show that the acquisition and loss of new genes appears to have been a central theme of rhabdovirus evolution, and has been associated with the appearance of alternative, overlapping and consecutive ORFs within the major structural protein genes, and the insertion and loss of additional ORFs in each gene junction in a clade-specific manner. Changes in the lengths of gene junctions accounted for as much as 48.5% of the variation in genome size from the smallest to the largest genome, and the frequency with which new ORFs were observed increased in the 3' to 5' direction along the genome. We also identify several new families of accessory genes encoded in these regions, and show that non-canonical expression strategies involving TURBS-like termination-reinitiation, ribosomal frame-shifts and leaky ribosomal scanning appear to be common. We conclude that rhabdoviruses have an unusual capacity for genomic plasticity that may be linked to their discontinuous transcription strategy from the negative-sense single-stranded RNA genome, and propose a model that accounts for the regular occurrence of genome expansion and contraction throughout the evolution of the Rhabdoviridae.