Ultrasound carotid artery intima-media thickness assessment for progression of atherosclerosis in lipid intervention studies.
Atherosclerotic cardiovascular disease is the number one cause of death in Western countries and is a rapidly growing problem worldwide. The majority of patients who present with acute coronary syndrome often have a low cardiovascular risk score, which is insufficient to warrant treatment prior to an acute coronary event. Alternatively, some high risk individuals never develop cardiovascular events. Identification of individuals who are truly at risk before an acute coronary event is critical. The long subclinical incubation period of atherosclerosis and the early involvement of the vessel wall in the disease provides an opportunity to evaluate the presence of atherosclerosis by imaging the arterial wall and to initiate treatment measures prior to an acute coronary event. The assessment of the thickness of the intima-media layer of the vessel wall, as well as detection of early plaques by ultrasound, is non-invasive and is the most sensitive, reliable, and safe method to detect those at risk. It is also the most validated method to detect progression of atherosclerosis. Evaluation of atherosclerosis progression by this technique has been used in large clinical trial settings, however, improvement in imaging and measurement technology may make this suitable for use in individual patients for therapeutic validity both in primary and secondary care settings.


Esophageal Carcinoma in Children: Report of 2 Cases and a Review of the Literature.
Esophageal carcinoma in children and adolescents is extremely rare. Here, we report 2 cases of pediatric esophageal carcinoma presenting with progressive dysphagia. There was not any underlying specific risk factor in our cases. The histopathological subtypes were adenocarcinoma in one and squamous cell carcinoma in another case. Response to combined modality treatment was good in the case of adenocarcinoma, while the patient with squamous cell carcinoma was unresponsive to treatment and died of the progressive disease. We reviewed the pediatric cases of esophageal carcinoma reported in the literature. Progressive dysphagia was observed in 89% of these cases. One third of pediatric cases had underlying risk factors. Squamous cell carcinoma is a more common type of childhood esophageal carcinoma. In contrast to adults, pediatric esophageal squamous cell carcinoma may distribute throughout the esophagus. Esophageal adenocarcinoma was seen in the distal esophagus in pediatric cases. Metastatic disease was found in 48% of pediatric patients at presentation, and the prognosis is poor. Collaborative efforts are needed for success in the treatment of esophageal carcinoma.


Antimicrobial prescription practices for outpatients with acute respiratory tract infections: A retrospective, multicenter, medical record-based study.
Antimicrobial stewardship for outpatients with acute respiratory tract infections (ARTIs) should be urgently promoted in this era of antimicrobial resistance. Previous large-sample studies were based on administrative data and had limited reliability. We aimed to identify current antimicrobial prescription practices for ARTIs by directly basing on medical records. This multicenter retrospective study was performed from January to December in 2018, at five medical institutes in Japan. We targeted outpatients aged ≥18 years whose medical records revealed International Classification of Diseases (ICD-10) codes suggesting ARTIs. We divided the eligible cases into three age groups (18-64 years, 65-74 years, and ≥75 years). We defined broad-spectrum antimicrobials as third-generation cephalosporins, macrolides, fluoroquinolones, and faropenem. Primary and secondary outcomes were defined as the proportion of antimicrobial prescriptions for the common cold and other respiratory tract infections, respectively. Totally, data of 3,940 patients were collected. Of 2,914 patients with the common cold, 369 (12.7%) were prescribed antimicrobials. Overall, compared to patients aged ≥75 years (8.5%), those aged 18-64 years (16.6%) and those aged 65-74 years (12.1%) were frequently prescribed antimicrobials for the common cold (odds ratio [95% confidential interval]; 2.15 [1.64-2.82] and 1.49 [1.06-2.09], respectively). However, when limited to cases with a valid diagnosis of the common cold by incorporating clinical data, no statistical difference was observed among the age groups. Broad-spectrum antimicrobials accounted for 90.2% of the antimicrobials used for the common cold. Of 1,026 patients with other respiratory infections, 1,018 (99.2%) were bronchitis, of which antimicrobials were prescribed in 49.9% of the cases. Broad-spectrum antimicrobials were the main agents prescribed, accounting for nearly 90% of prescriptions in all age groups. Our data suggested a favorable practice of antimicrobial prescription for outpatients with ARTIs in terms of prescribing proportions, or quantitative aspect. However, the prescriptions were biased towards broad-spectrum antimicrobials, highlighting the need for further antimicrobial stewardship in the outpatient setting from a qualitative perspective.


Elevated tryptase levels selectively cluster in myeloid neoplasms: a novel diagnostic approach and screen marker in clinical haematology.
Recent data suggest that tryptase, a mast cell enzyme, is expressed in neoplastic cells in myeloid leukaemias. In several of these patients, increased serum tryptase levels are detectable. We have determined serum tryptase levels in 914 patients with haematological malignancies, including myeloproliferative disorders (n = 156), myelodysplastic syndromes (MDS, n = 241), acute myeloid leukaemia (AML, n = 317), systemic mastocytosis (SM, n = 81), non-Hodgkin's lymphoma (n = 59) and acute lymphoblastic leukaemia (n = 26). Moreover, tryptase was measured in 136 patients with non-neoplastic haematological disorders, 102 with non-haematological disorders and 164 healthy subjects. In healthy subjects, the median serum tryptase was 5.2 ng mL(-1). Elevated serum tryptase levels were found to cluster in myeloid neoplasm, whereas almost all patients with lymphoid neoplasms exhibited normal tryptase. Among myeloid neoplasms, elevated tryptase levels (> 15 ng mL(-1)) were recorded in > 90% of patients with SM, 38% with AML, 34% with CML and 25% with MDS. The highest tryptase levels, often > 1000 ng mL(-1), were found in advanced SM and core-binding-factor leukaemias. In most patients with non-neoplastic haematological disorders and non-haematological disorders analysed in our study, tryptase levels were normal, the exception being a few patients with end-stage kidney disease and helminth infections, in whom a slightly elevated tryptase was found. In summary, tryptase is a new diagnostic marker of myeloid neoplasms and a useful test in clinical haematology.


[Kinetics of dissolution of solid protein substrates by proteinases. Selection of the reaction mechanism].
Experimental data on the rate of enzymatic degradation of a solid protein phase in relation to reagent concentrations in the gelatin gel--proteolytic enzyme solution system was obtained. It was shown that different transformations of the Michaelis-Menten equation do not provide any satisfactory description of the reaction kinetics. A method for the discrimination of putative reaction mechanisms based on the analysis of effective values of constants in the general equation for the rates of dissolution of proteins with different molecular weights was proposed. The experimental results are in favour of a two-step consecutive mechanism taking account of surface concentrations of the adsorbed enzyme and hydrolyzable bonds.


[Adaptation of sensory systems of gamasid mites (Parasitiformes, Gamasina) to dwelling in different environment].
The main complication sensory organs (the palpal organ and the tarsal sensory complex) of several species of gamasid mites were studied in scanning electron microscope. The species examined included permanent ectoparasites (Laelaps agilis, Laelaptidae), parasites of the nasal cavity and respiratory tract of birds (Sternostoma tracheocolum and Ptilonyssus reguli, Rhinonyssidae), dwellers of the sea littoral zone (Parasitus kempersi, Parasitus immanis, Parasitidae), and mites found on soil and on plants (Amblyseius barkeri, Parasitidae). Similar sensillar types, including olfactory SW-WP sensilla, contact chemo-mechanosensory (SW-UP and DW-UP) sensilla, termo-chemo-mechanosensitive (DW-WP) sensilla of two types, and tactile (NP) sensilla were found in all these species, excluding endoparasites, where some sensillar types (in particular, DW-WP sensilla with slit-like pores) are absent. It was shown that the topography of olfactory SW-WP sensilla of the tarsal complex reflects taxonomic position and phylogenetic history of mite genera, whereas the number of certain sensillar types and the degree of their development reflect ecological specialization of species. The palpal organ is characterized by rather uniform structure in mites of different families, dwellers of different environments, except for the endoparasites of the family Rhinonyssidae, where this organ is strongly reduced.


Evaluation of a PSMA-targeted BNF nanoparticle construct.
Early detection enables improved prognosis for prostate cancer (PCa). A promising target for imaging and therapy of PCa is the prostate-specific membrane antigen (PSMA), which exhibits both expression within the epithelium of PCa cells, and becomes internalized upon ligand binding. Here we report the synthesis of a PSMA-targeted bionized nanoferrite (BNF) nanoparticle and its biological evaluation in an experimental model of PCa. The BNF nanoparticle formulation exhibits properties conducive to targeted imaging such as stealth, prolonged circulation time and enhanced clearance from non-target sites. Optical imaging of the targeted BNF in vivo indicates preferential accumulation in PSMA+ tumors 4 h post-injection, suggesting target specificity. On the other hand, non-targeted nanoparticles exhibit lower uptake with similar accumulation in both PSMA+ and PSMA- tumors indicating tumor access without preferential accumulation. Imaging with single photon emission computed tomography (SPECT) and biodistribution studies of a modified construct indicate highest tumor accumulation at 48 h post-injection [4.3 ± 0.4 percentage injected dose per gram of tissue (%ID g(-1))], with tumor/blood and tumor/muscle ratios of 7.5 ± 2.4 and 11.6 ± 1.2 %ID g(-1), respectively. Ex vivo fluorescence microscopy, Prussian blue staining, immunohistochemistry and biodistribution studies confirm enhanced nanoparticle uptake in PSMA+ tumors compared to those not expressing PSMA. The BNF nano-formulation described is promising for PSMA-targeted imaging applications in vivo.


A multi-criteria approach to Great Barrier Reef catchment (Queensland, Australia) diffuse-source pollution problem.
This paper presents a multi-criteria based tool for assessing the relative impact of diffuse-source pollution to the Great Barrier Reef (GBR) from the river basins draining into the GBR lagoon. The assessment integrates biophysical and ecological data of water quality and pollutant concentrations with socio-economic information pertaining to non-point source pollution and (potential) pollutant impact. The tool generates scores for each river basin against four criteria, thus profiling the basins and enabling prioritization of management alternatives between and within basins. The results support policy development for pollution control through community participation, scientific data integration and expert knowledge contributed by people from across the catchment. The results specifically provided support for the Reef Water Quality Protection Plan, released in October 2003. The aim of the plan is to provide a framework for reducing discharge of sediment, nutrient and other diffuse-source loads and (potential) impact of that discharge and for prioritising management actions both between and within river basins.


[Digestive forms of Chagas disease and carcinogenesis: a study of association].
The authors analyze the relation between gastrointestinal carcinogenesis and Chagas disease, based on detailed review of the literature. To this end, epidemiological, experimental and human material pathology description studies have been selected. The article discusses the possibility of protection being afforded by not fully known morphokinetic cellular, immune and neuroendocrine factors that would be secondary to plexus degeneration. Also aspects related to the parasite-host interaction from the viewpoint of epithelial modulation of colonic mucosa and its antitumor implications are presented. Finally, it exposes the pathophysiological mechanism of esophageal cancer development in patients with mega-organ. In conclusion, chagasic colopathy, especially the intrinsic neuronal damage, is a study model that can contribute to the understanding of colorectal carcinogenesis.


The inhibition of human platelet function by ganodermic acids.
Human gel-filtered platelets aggregate at greater than 20 microM-ganodermic acid S [lanosta-7,9(11),24-triene-3 beta, 15 alpha-diacetoxy-26-oic acid] [Wang, Chen, Shiao & Wang (1989) Biochim. Biophys. Acta 986, 151-160]. This study showed that platelets at less than 20 microM-ganodermic acid S displayed both concentration- and time-dependent inhibition of function, in which the agent potency in response to inducers was ADP-fibrinogen greater than collagen greater than thrombin. The agent caused a biphasic time-dependent effect on platelet phosphoinositide metabolism. The first phase involved the decrease in the pool size of phosphoinositide by 10-20%. The second phase, in which both the resynthesis of phosphatidylinositol 4,5-bisphosphate (PIP2) and the decrease of [32P]phosphatidic acid occurred, took place after 30 min. Scanning electron microscopy also revealed a time-dependent morphological change in platelets in the presence of the agent. The cells initially became spiculate discs, then swelled to a 'potato-like' morphology at 60 min. Further studies on the time-dependent inhibition of thrombin response revealed that: (1) the percentage inhibition of cell aggregation was comparable with that occurring with an increase of cytosolic free Ca2+ concentration [( Ca2+]i) or the phosphorylation of marker proteins; (2) [32P]Pi-labelled platelets showed the time-dependent inhibition of thrombin-stimulated PIP2 resynthesis as indicated by first-2-min time-course studies of phosphoinositide interconversion; (3) scanning electron microscopy revealed that the aged platelet population showed an increase in the percentage of non-responding cells on prolonged incubation. The results, taken together, enabled one to discuss a possible mechanism for the time-dependent inhibition by ganodermic acid S of platelet response to thrombin.


Simultaneous automatic control of oxygen and carbon dioxide blood gases during cardiopulmonary bypass.
In this work an automatic control strategy is presented for the simultaneous control of oxygen and carbon dioxide blood gas partial pressures to be used during cardiopulmonary bypass surgery with heart-lung machine support. As the exchange of blood gases in the artificial extracorporeal lung is a highly nonlinear process comprising varying time delays, uncertainties, and time-varying parameters, it is currently being controlled manually by specially trained perfusionist staff. The new control strategy includes a feedback linearization routine with augmented time-delay compensation and two external linear gain-scheduled controllers, for partial oxygen and carbon dioxide pressures. The controllers were robustly tuned and tested in simulations with a detailed artificial lung (oxygenator) model in cardiopulmonary bypass conditions. Furthermore, the controllers were implemented in an ex vivo experiment using fresh porcine blood as a substitute fluid and a special deoxygenation technique to simulate a patient undergoing cardiopulmonary bypass. Both controllers showed robust stability during the experiments and a good disturbance rejection to extracorporeal blood flow changes. This automatic control strategy is proposed to improve patient's safety by fast control reference tracking and good disturbance rejection under varying conditions.


Substance use disorders: leading the road to recovery.
Substance use disorders are diseases of the brain that create a dependency on drug(s), either prescription or illicit. These diseases affect millions of people worldwide, yet, there are few treatments that can help patients in the long term. This opinion piece looks at strategies researchers and institutes are taking to help find treatments as well as at new therapies in clinical trials. It provides an outlook on how a changing public perspective of these diseases can ultimately lead to a brighter outlook for substance use disorder treatments.


[The clinical evaluation of a change in lipid peroxidation in patients with ischemic heart disease and cardiac arrhythmias].
Overall 93 patients suffering from coronary heart disease /CHD/ (without acute myocardial infarction) complicated by cardiac rhythm impairment were followed up. Investigation of lipid peroxidation (LPO) and of the antioxidant system (AOS) in those patients revealed LPO activation and a decrease of the AOS activity both at the moment of arrhythmia and after its arrest, with the highest LPO activation being seen in the hyper- and hypokinetic hemodynamic variants. The experimental studies have demonstrated a reverse relationship between the rise of LPO products in the blood, LPO activation in myocytes and the onset of arrhythmia because of an increase in the density of Na(+)-Ca++ channels of the membrane, accumulation of those ions by the cell, and an increase of the transmembranous potential at rest. Therefore, to prevent and treat cardiac rhythm impairment in CHD patients, it is necessary to administer the blockers of Na(+)-Ca++ channels (etmozin, ethacizine, verapamil and so forth) and antioxidants that raise the reserve capacity of the AOS, including laser therapy.


Ultrasensitive detection of genetic variation based on dual signal amplification assisted by isothermal amplification and cobalt oxyhydroxide nanosheets/quantum dots.
PML/RARα fusion gene (P/R) is the characteristic signature genetic variation of acute promyelocytic leukemia (APL). Here, by integrating triple-stranded DNA hybridization-triggered strand displacement amplification (tri-HT SDA) and cobalt oxyhydroxide nanosheets/quantum dots (CoOOH/QD)-based amplification, we constructed a novel biosensor of easy-operating, time-saving and high sensitivity for detecting P/R to meet clinical needs. Owing to the specific recognition and efficient amplification of tri-HT SDA as well as impressive anti-interference and considerable amplification of CoOOH/QD, this biosensor demonstrated a wide dynamic range (10 fM to 10 nM) with a low limit of detection (5.50 fM) in P/R detection. Additionally, this biosensor could detect P/R spiked into human serum with good recoveries and relative standard deviation (RSD), thus potentially exhibiting ultrasensitive and specific nuclear acid sequence detection ability in clinical diagnosis owing to combing isothermal amplification and nanomaterials.


Viral replication-independent blockade of dendritic cell maturation and interleukin-12 production by human herpesvirus 6.
Human herpesvirus 6 (HHV-6) is a potentially immunosuppressive CD4(+)-T-lymphotropic betaherpesvirus that causes severe human thymocyte depletion in heterochimeric SCID-hu thy/liv mice and has been implicated as a potential cofactor in the progression of AIDS. However, the mechanisms of HHV-6-mediated immunosuppression have not yet been fully elucidated. We investigated the phenotypic and functional alterations induced by HHV-6 on peripheral blood-derived human dendritic cells (DC). The infection of DC with HHV-6 A or B was nonproductive, as revealed by calibrated real-time PCR measuring the accumulation of viral genome equivalents over time. Nevertheless, preexposure to HHV-6 markedly impaired the maturation of DC driven by gamma interferon and lipopolysaccharide, as shown by the reduced surface expression of major histocompatibility complex class I molecules, HLA-DR, CD40, and CD80. Moreover, HHV-6, but not the closely related betaherpesvirus HHV-7, dramatically suppressed the secretion of interleukin-12 (IL-12) p70 by DC, while the production of other cytokines that influence DC maturation, i.e., IL-10 and tumor necrosis factor alpha, was not significantly modified. Likewise, the secretion of the CC chemokines macrophage inflammatory protein 1beta and RANTES was unaltered. Functionally, a pretreatment with HHV-6 impaired the ability of DC to stimulate allogeneic T-cell proliferation. Altogether, these data identify interference with the functional maturation of DC as a potential mechanism of HHV-6-mediated immunosuppression.


Prediction of Response in Head and Neck Tumor: Focus on Main Hot Topics in Research.
Radiation therapy is a cornerstone in the treatment of head and neck cancer patients; actually, their management is based on clinical and radiological staging with all patients at the same stage treated in the same way. Recently the increasing knowledge in molecular characterization of head and neck cancer opens the way for a more tailored treatment. Patient outcomes could be improved by a personalized radiotherapy beyond technological and anatomical precision. Several tumor markers are under evaluation to understand their possible prognostic or predictive value. In this paper we discuss those markers specific for evaluate response to radiation therapy in head and neck cancer for a shift toward a biological personalization of radiotherapy.


Phonons of the anomalous element cerium.
Many physical and chemical properties of the light rare-earths and actinides are governed by the active role of f electrons, and despite intensive efforts the details of the mechanisms of phase stability and transformation are not fully understood. A prominent example which has attracted a lot of interest, both experimentally and theoretically over the years is the isostructural γ - α transition in cerium. We have determined by inelastic X-ray scattering, the complete phonon dispersion scheme of elemental cerium across the γ → α transition, and compared it with theoretical results using ab initio lattice dynamics. Several phonon branches show strong changes in the dispersion shape, indicating large modifications in the interactions between phonons and conduction electrons. This is reflected as well by the lattice Grüneisen parameters, particularly around the X point. We derive a vibrational entropy change ΔS(γ-α)(vib) ≈ (0.33+/-0.03)k(B), illustrating the importance of the lattice contribution to the transition. Additionally, we compare first principles calculations with the experiments to shed light on the mechanism underlying the isostructural volume collapse in cerium under pressure.


Percutaneously inserted long-term central venous catheters in pigs of different sizes.
Pigs are used for long-term biomedical experiments requiring repeated injections, infusions and collections of blood samples. Thus, it is necessary for vascular catheters to be indwelling to avoid undue stress to the animals and the use of restraints. We propose a refined model of percutaneous insertion of long-term central venous catheters to minimize the surgical trauma and postoperative complications associated with catheter insertion. Different sizes of needles (18 Ga versus 21 Ga) for initial puncture of the veins were compared. In conventional pigs weighing less than 30 kg, catheter insertion may be facilitated by using a microintroducer set with a 21 Ga needle. In pigs weighing 50 kg, a standard 18 Ga needle may be preferable.


Delineation of Novel Compound Heterozygous Variants in LTBP2 Associated with Juvenile Open Angle Glaucoma.
Juvenile open angle glaucoma (JOAG), which is an uncommon form of primary open angle glaucoma, is a clinically and genetically heterogeneous disorder. We report on a family with a recessively inherited form of JOAG. The proband has a superior and an inferior never fiber layer thinning in both the eyes and the nasal visual field (VF) defects in the left eye, which are clinical findings consistent with glaucomatous optic neuropathy. Whole exome sequencing revealed two novel compound heterozygous variants [c.2966C>G, p.(Pro989Arg); c.5235T>G, p.(Asn1745Lys)] in latent transforming growth factor-beta-binding protein 2 (LTBP2) segregating with the phenotype. Both these variants are predicted to replace evolutionary conserved amino acids, have a pathogenic effect on the encode protein, and have very low frequencies in the control databases. Mutations in LTBP2 are known to cause the Weill-Marchesani syndrome and a Weill-Marchesani-like syndrome, which include glaucoma in their clinical presentation. However, to our knowledge, this is the first published case of a JOAG subject associated with recessively inherited variants of LTPB2 and, thus, expands the repertoire of the known genetic causes of JOAG and the phenotypic spectrum of LTBP2 alleles.


The recruitment and participation of Hispanic women in nursing research: a learning process.
Limited information is available regarding the selection of culturally appropriate methods for obtaining data from the Hispanic population, which is currently the largest and fastest growing ethnic minority in the United States. Due to this lack of knowledge which would guide the investigator in the recruitment and retention of Hispanics in research studies, the Hispanic population remains understudied and underserved. This article describes the numerous challenges that were encountered in the actual process of recruiting Hispanic women into a research study which examined adjustment among breast cancer survivors. The rate of refusal to participate was unexpectedly high, which necessitated additional recruitment strategies, including a renewed emphasis on the importance of the study and the option of meeting with other members of the community in a group. Women who refused to participate in the study stated concerns related to a loss of health benefits, the influence of family members, and a recurrence of emotional stress. Instrument issues are also addressed. This learning process of developing, refining, and testing effective methods for recruiting Hispanic women in research studies remains an ongoing endeavor; through these efforts it is hoped that culturally competent health care can become a reality.


Studies on the SV40-like papovavirus SV40-GBM. I. Genomic analysis by restriction endonucleases and electron microscopy after propagation in CV-1 monkey cells.
Infection of CV-1 monkey cells with SV40-GBM, a papovavirus isolated from a human glioblastoma multiforme, resulted in the appearance of defective viral DNA molecules. In contrast to SV40 wild-type, two main types of variant DNA molecules could be found after three viral passages at multiplicities of infection of about 10. The molecules of one variant DNA (GBM3-L) were about 19% shorter than the GBM3-H DNA molecules and the DNA of the original GBM isolate, as demonstrated by electron microscopy. Restriction enzyme analysis revealed that GBM3-L DNA had lost both the EcoRI and the HpaII cleavage sites which are located in the late viral genome region. Furthermore, SV40 GBM3-L did not possess the two PvuII sites which are located in the late genome region, and a portion of the GBM3-H and GBM3-L DNA molecules had lost the unique KpnI site. Heteroduplex analysis verified that the rearrangements in the GBM3-L DNA are located only in the late region of this DNA. The possible differences between SV40 wild-type and SV40-GBM are discussed on the basis of these results.


Immobilization of Papain on Chitin and Chitosan and Recycling of Soluble Enzyme for Deflocculation of Saccharomyces cerevisiae from Bioethanol Distilleries.
Yeast flocculation (Saccharomyces cerevisiae) is one of the most important problems in fuel ethanol production. Yeast flocculation causes operational difficulties and increase in the ethanol cost. Proteolytic enzymes can solve this problem since it does not depend on these changes. The recycling of soluble papain and the immobilization of this enzyme on chitin or chitosan were studied. Some cross-linking agents were evaluated in the action of proteolytic activity of papain. The glutaraldehyde (0.1-10% w·v(-1)), polyethyleneimine (0.5% v·v(-1)), and tripolyphosphate (1-10% w·v(-1)) inactivated the enzyme in this range, respectively. Glutaraldehyde inhibited all treatments of papain immobilization. The chitosan cross-linked with TPP in 5 h of reaction showed the yield of active immobilized enzyme of 15.7% and 6.07% in chitosan treated with 0.1% PEI. Although these immobilizations have been possible, these levels have not been enough to cause deflocculation of yeast cells. Free enzyme was efficient for yeast deflocculation in dosages of 3 to 4 g·L(-1). Recycling of soluble papain by centrifugation was effective for 14 cycles with yeast suspension in time perfectly compatible to industrial conditions. The reuse of proteases applied after yeast suspension by additional yeast centrifugation could be an alternative to cost reduction of these enzymes.


Differential introgression reveals candidate genes for selection across a spruce (Picea sitchensis × P. glauca) hybrid zone.
Differential patterns of introgression between species across ecological gradients provide a fine-scale depiction of extrinsic and intrinsic factors that contribute to the maintenance of species barriers and adaptation across heterogeneous environments. Introgression was examined for 721 individuals collected from the ecological transition zone spanning maritime to continental climates within the Picea sitchensis-Picea glauca contact zone using a panel of 268 candidate gene single nucleotide polymorphisms. Geographic clines showed a strong spatial relationship between allele frequencies and both distance from the ocean along major rivers and mean annual precipitation, indicating a strong role for environmental selection. Interspecific patterns of differentiation using outlier tests revealed three candidate genes that may be targets of long-term divergent selection between the parental species, although contemporary genomic clines within the hybrid zone suggested neutral patterns of introgression for these genes. This study provides a fine-scale analysis of locus-specific introgression, identifying a suite of candidate loci that may be targets of extrinsic or intrinsic selection, with broad application in understanding local adaptation to climate.


Large, rapidly evolving gene families are at the forefront of host-parasite interactions in Apicomplexa.
The Apicomplexa is a phylum of parasitic protozoa, which includes the malaria parasite Plasmodium, amongst other species that can devastate human and animal health. The past decade has seen the release of genome sequences for many of the most important apicomplexan species, providing an excellent basis for improving our understanding of their biology. One of the key features of each genome is a unique set of large, variant gene families. Although closely related species share the same families, even different types of malaria parasite have distinct families. In some species they tend to be found at the ends of chromosomes, which may facilitate aspects of gene expression regulation and generation of sequence diversity. In others they are scattered apparently randomly across chromosomes. For some families there is evidence they are involved in antigenic variation, immune regulation and immune evasion. For others there are no known functions. Even where function is unknown these families are most often predicted to be exposed to the host, contain much sequence diversity and evolve rapidly. Based on these properties it is clear that they are at the forefront of host-parasite interactions. In this review I compare and contrast the genomic context, gene structure, gene expression, protein localization and function of these families across different species.


Properties of six pesticide degradation plasmids isolated from Alcaligenes paradoxus and Alcaligenes eutrophus.
Biophysical and genetic properties of six independently isolated plasmids encoding the degradation of the herbicides 2,4-dichlorophenoxyacetic acid and 4-chloro-2-methylphenoxyacetic acid are described. Four of the plasmids, pJP3, pJP4, pJP5, and pJP7, had molecular masses of 51 megadaltons, belonged to the IncP1 incompatibility group, and transferred freely to strains of Escherichia coli, Rhodopseudomonas sphaeroides, Rhizobium sp., Agrobacterium tumefaciens, Pseudomonas putida, Pseudomonas fluorescens, and Acinetobacter calcoaceticus. In addition, these four plasmids conferred resistance to merbromin, phenylmercury acetate, and mercuric ions, had almost identical restriction endonuclease cleavage patterns, and encoded degradation of m-chlorobenzoate. The two other plasmids, pJP2 and pJP9, did not belong to the IncP1 incompatibility group, had molecular masses of 37 megadaltons, encoded the degradation of phenoxyacetic acid, and possessed identical restriction endonuclease cleavage patterns.


[Interspecies chimeras of mammals].
Chimaeras between closely related and distant mammalian species were obtained during the last decade using the aggregation and injection methods. In the chimaeras between the closely related species Mus musculus--M. caroli, the cell clones of both parental species are equally represented; such chimaeras are fertile and their development does not differ from that of intraspecific chimaeras. But in the chimaeras between distant species mouse--rat or sheep--goat a selective predominance of cells with one genotype over those with another was noted. This leads to the total elimination of cell populations with one genotype, rat in particular in the mouse--rat chimaeras, or, possibly, to the sterility of the sheep--goat chimaeras.


Profiling joint tissues at single-cell resolution: advances and insights.
Advances in the profiling of human joint tissues at single-cell resolution have provided unique insights into the organization and function of these tissues in health and disease. Data generated by various single-cell technologies, including single-cell RNA sequencing and cytometry by time-of-flight, have identified the distinct subpopulations that constitute these tissues. These timely studies have provided the building blocks for the construction of single-cell atlases of joint tissues including cartilage, bone and synovium, leading to the identification of developmental trajectories, deciphering of crosstalk between cells and discovery of rare populations such as stem and progenitor cells. In addition, these studies have revealed unique pathogenetic populations that are potential therapeutic targets. The use of these approaches in synovial tissues has helped to identify how distinct cell subpopulations can orchestrate disease initiation and progression and be responsible for distinct pathological outcomes. Additionally, repair of tissues such as cartilage and meniscus remains an unmet medical need, and single-cell methodologies can be invaluable in providing a blueprint for both effective tissue-engineering strategies and therapeutic interventions for chronic joint diseases such as osteoarthritis and rheumatoid arthritis.


Hemoglobin synthesis in murine virus-induced leukemic cells in vitro: stimulation of erythroid differentiation by dimethyl sulfoxide.
Cells of a cloned line of murine virus-induced erythroleukemia were stimulated to differentiate along the erythroid pathway by dimethyl sulfoxide at concentrations that did not inhibit growth. A rise in the number of benzidine-positive normoblasts was accompanied by increased synthesis of heme and hemoglobin and a decrease in the malignancy of the cells. This action of dimethyl sulfoxide, which was reversible, may represent the derepression of leukemic cells to permit their maturation.


Mechanisms of tyrosine hydroxylase regulation during pregnancy: evidence for protein dephosphorylation during the prolactin surges.
This study examined tyrosine hydroxylase (TH) activity in the stalk-median eminence (SME) and TH messenger RNA (mRNA) signal levels in the arcuate nuclei of the hypothalamus during early, middle, and late pregnancy and related these to circulating levels of ovarian steroids. In addition, this study evaluated the intracellular mechanism(s) which contributes to the semicircadian rhythm in tuberoinfundibular dopaminergic neuronal activity during early pregnancy. The catalytic activity of TH in the SME was determined from the in vitro rate of 3,4,dihydroxyphenylalanine accumulation after inhibiting DOPA decarboxylase with brocresine. TH mRNA signal levels were evaluated by in situ hybridization. TH mRNA signal levels in the arcuate nuclei were 30% lower at 1000 h on day 20 of pregnancy as compared to days 7 and 11, whereas TH activity in the SME at 1000 h was not significantly different on days 7, 11, 16, and 20. Serum PRL levels were low (3-6 ng/ml) and unchanged at 1000 h on days 7, 11, 16, and 20. Circulating progesterone levels increased from 111 to 191 ng/ml on days 7 and 16, respectively, and then declined to 69 ng/ml by day 20. Serum estradiol levels increased from 38 to 106 pg/ml on day 7 and 16, respectively, and then remained elevated on day 20. Thus, the reduction in TH mRNA signal levels during late pregnancy is temporally related to the increased estradiol/progesterone ratio. Elevated serum PRL levels at 0330 h and 1800 h on day 7 were characteristic of the nocturnal and diurnal PRL surges of early pregnancy. Circulating PRL levels were low during the intersurge times (2330 and 1000 h) on day 7 and at all times examined on day 11. TH activity in the SME on day 7 was lower during the PRL surges as compared to the intersurge times, whereas TH activity on day 11 was similar at all times and comparable to the intersurge levels of early pregnancy. Okadaic acid, a protein phosphatase inhibitor, reversed the reduction in TH activity during the nocturnal and diurnal PRL surges, but did not significantly alter TH activity during the intersurge period on day 7. TH mRNA signal levels in the arcuate nuclei were similar throughout day 7. These data indicate that protein dephosphorylation, but not changes in the TH gene expression, may contribute to the semicircadian rhythm in TH activity during early pregnancy.


Atmospheric CO2 Concentration and N Availability Affect the Balance of the Two Photosystems in Mature Leaves of Rice Plants Grown at a Free-Air CO2 Enrichment Site.
Atmospheric CO2 concentration ([CO2]) has been substantially increasing. Responses of leaf photosynthesis to elevated [CO2] have been intensively investigated because leaf photosynthesis is one of the most important determinants of crop yield. The responses of photosynthesis to elevated [CO2] can depend on nitrogen (N) availability. Here, we aimed to investigate the significance of the appropriate balance between two photosystems [photosystem I (PSI) and photosystem II (PSII)] under various [CO2] and N levels, and thus to clarify if responses of photosynthetic electron transport rates (ETRs) of the two photosystems to elevated [CO2] are altered by N availability. Thus, we examined parameters of the two photosystems in mature leaves of rice plants grown under two [CO2] levels (ambient and 200 μmol mol-1 above ambient) and three N fertilization levels at the Tsukuba free-air CO2 enrichment experimental facility in Japan. Responses of ETR of PSII (ETRII) and ETR of PSI (ETRI) to [CO2] levels differed among N levels. When moderate levels of N were applied (MN), ETRI was higher under elevated [CO2], whereas at high levels of N were applied (HN), both ETRII and ETRI were lower under elevated [CO2] compared with ambient [CO2]. Under HN, the decreases in ETRII and ETRI under elevated [CO2] were due to increases in the non-photochemical quenching of PSII [Y(NPQ)] and the donor side limitation of PSI [Y(ND)], respectively. The relationship between the effective quantum yields of PSI [Y(I)] and PSII [Y(II)] changed under elevated [CO2] and low levels of N (LN). Under both conditions, the ratio of Y(I) to Y(II) was higher than under other conditions. The elevated [CO2] and low N changed the balance of the two photosystems. This change may be important because it can induce the cyclic electron flow around PSI, leading to induction of non-photochemical quenching to avoid photoinhibition.


The mobility of mitochondria: Intercellular trafficking in health and disease.
The view that genes are constrained within somatic cells is challenged by in vitro evidence, and more recently by in vivo studies which demonstrate that mitochondria with their mitochondrial DNA (mtDNA) payload not only can, but do move between cells in tumour models and in mouse models of tissue damage. Using mouse tumour cell models without mtDNA to reflect mtDNA damage, we have shown that these cells grow tumours only after acquiring mtDNA from cells in the local microenvironment resulting in respiration recovery, tumorigenesis and metastasis. Mitochondrial transfer between cells has also been demonstrated following ischaemia-induced injury in the heart and brain and in lung epithelium, and following lung inflammation. In vitro investigations suggest that stem cells may be mitochondrial donors. The ability of mitochondria to move between cells appears to be an evolutionarily-conserved phenomenon, relevant to diseases with compromised mitochondrial function including neurodegenerative, neuromuscular and cardiovascular diseases as well as cancer and ageing.


Irys Extract.
Irys Extract is a software tool for generating genomic information from data collected by the BioNano Genomics Irys platform. The tool allows the user easy access to the raw data in the form of cropped images and genetically aligned intensity profiles. The latter are also made compatible with the BED format for using with popular genomic browsers such as the UCSC Genome Browser. Irys Extract has been developed in Matlab R2015a, it was tested to work with IrysView 22.214.171.12479 and AutoDetect 188.8.131.5259, and it currently runs under Microsoft Windows operating systems (7-10). Irys Extract can be downloaded alongside its manual and a demo dataset at http://www.nanobiophotonix.com and https://sites.google.com/site/raniarielly/. firstname.lastname@example.com. Supplementary data are available at Bioinformatics online.


Lenalidomide in DLBCL: are we past the cell of origin?
Single-agent lenalidomide has modest activity in diffuse large B-cell lymphoma (DLBCL) and is thought to be more potent in activated B-cell (ABC) lymphomas, which are more treatment-resistant. However, the addition of lenalidomide to rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) in randomized clinical trials has shown equivocal benefit, despite phase 2 studies that suggested otherwise. These equivocal results suggest that either the cell of origin (COO) has limited importance for prescribing lenalidomide, or that lenalidomide is not the optimal agent for exploiting the vulnerability of ABC lymphomas. As more recent analyses have shown that the genetic landscape of DLBCL is considerably more complex than the binary COO paradigm, the disappointing impact of lenalidomide is less surprising. In contrast to the marginal benefit from the addition of lenalidomide to R-CHOP, recent studies suggest that lenalidomide in combination with novel agents has potent activity. Lenalidomide was recently approved in combination with the anti-monoclonal B-cell antibody tafasitamab for patients with relapsed DLBCL after 1 to 3 previous treatments. This combination has led to surprisingly prolonged progression-free survival rates, along with possible cure in a subset of patients. In addition, early-phase single-arm trials are also showing deep and durable responses in relapsed patients when lenalidomide is combined with the novel agents ibrutinib and venetoclax. Although these drugs have limited single-agent activity in DLBCL, their pronounced activity in combination suggests a possible unique synergistic effect. Overall, recent studies suggest that lenalidomide will continue to be an active player in the treatment for DLBCL but likely in combination with other novel agents rather than in combination with chemotherapy.


Side-to-end colorectal anastomosis with monofilament stainless steel wire.
Anastomotic separation after anterior resection of the rectum remains a major problem, although the frequency of anastomotic dehiscence after anterior resection varies. Monofilament stainless steel wire suture is an inert suture, has excellent holding power, and is associated with a low probability of wound infection. Side-to-end coloproctostomy for anastomotic reconstruction after anterior resection has proved to be a satisfactory alternative to the usual end-to-end anastomosis. A combination of wire suture with side-to-end coloproctostomy was performed in 60 consecutive anterior resections in which there were two clinical anastomotic leaks and no deaths attributable to anastomotic dehiscence. Simultaneous loop transverse colostomy was performed in 13 cases (22%), and all colostomies were subsequently closed without difficulty. This technique is recommended for reconstruction of bowel continuity after anterior resection of the rectum.


[Growth of Artenia franciscana and A. persimilis (Crustacea, Anostraca) under controlled conditions].
Growth (length) was compared in A. franciscana populations from Utah, United States of America, and Yape, Chile, and A. persimilis (population from Salinas de Hidalgo, Argentina), cultured under controlled conditions (24 degrees C, seawater 35 g/l, fed with Chaetoceros calcitrans). There were significant differences among all stages of A. franciscana (Utah) and A. persimilis, whereas we found significant differences only in two stages of A. persimilis and A. franciscana (Yape); both A. franciscana had similar total lengths in all stages analyzed. Habitat adaptations could explain these results.


Rapid pituitary tumor shrinkage with dissociation between antiproliferative and antisecretory effects of a long-acting octreotide in an acromegalic patient.
Criteria to define the response to somatostatin (SS) analogs (SSA) in acromegaly are based on biochemical control of the disease. However, the mechanisms of action of SSAs in inhibiting tumor growth and hormonal secretion are only partially understood, and the two effects may occur independently. The objective of the study was to investigate the dissociation between antiproliferative and antisecretive effects of SSA in an octreotide-resistant patient displaying dramatic tumor shrinkage during primary therapy with octreotide LAR. We characterized somatostatin and dopamine D(2) receptor expression by immunohistochemistry and real-time RT-PCR. The effects of different receptor-selective, bispecific analogs, and chimeric somatostatin/dopamine compounds on GH secretion and cell proliferation in primary cell cultures of the tumor were assessed. The expression of SS receptor subtypes (sst)(5) and D(2) receptor was higher, compared with the other receptor subtypes. GH inhibition by SS-14 and the two chimeric somatostatin/dopamine compounds was scant but greater than subtype-selective and sst(2)/sst(5) bispecific agonists. Conversely, cell growth was potently inhibited by all test substances. However, SS-14, sst(2)/sst(5) bispecific agonist, and chimeric molecules were more potent than the other compounds. The significant antiproliferative effect of octreotide seems to be related to the higher expression of sst(5) and the negligible antihormonal effect to the lower expression of sst(2). However, activation of multiple receptors by new analogs may produce better control of tumor cell activities. The dissociation between antisecretive and antiproliferative effects observed in vivo and in vitro confirms that SSAs may induce tumor shrinkage despite the lack of effect on GH secretion.


Th1 and th2 cytokine expression in nasopharyngeal secretions during acute bronchiolitis in children younger than two years old.
We characterised the T helper cytokine profiles on the surface of nasal mucosa of children with acute bronchiolitis caused by Respiratory Syncytial Virus, Parainfluenza Virus, Influenza Virus, Adenovirus, or without any viral identification, in order to examine whether these viral types modified cytokine responses. As an additional objective we sought to determine if T helper polarisation was associated with other demographic and environmental factors. A prospective study of children with acute bronchiolitis was performed. Patients were recruited from the emergency department of a central hospital in Lisbon, Portugal. Demographical, epidemiological and clinical data were gathered from a questionnaire. Nasal swabs were collected for viral studies (immunofluorescence) and T cell cytokine responses (detection of expression of interleukins 4, 13, 12 and interferon-γ by real-time polymerase chain reaction assays). Respiratory Syncytial Virus elicited lower levels of interleukin 4, when compared with samples without virus identification. A similar tendency to Th1 polarisation was found in older children, in those who attended day-care centres, and in breastfed individuals. Exposure to tobacco smoke was associated with a Th2 bias in this population. These findings show that Respiratory Syncytial Virus infection contributes to Th1 polarisation in immune response of respiratory mucosa, an effect that is similar to other environmental factors. Further studies are needed to assess immune response to other infectious causes of acute bronchiolitis.


Minimum average correlation energy filters.
The synthesis of a new category of spatial filters that produces sharp output correlation peaks with controlled peak values is considered. The sharp nature of the correlation peak is the major feature emphasized, since it facilitates target detection. Since these filters minimize the average correlation plane energy as the first step in filter synthesis, we refer to them as minimum average correlation energy filters. Experimental laboratory results from optical implementation of the filters are also presented and discussed.


Prediction and risk of dysphagia after uvulopalatopharyngoplasty and uvulopalatoplasty.
To test the hypothesis that preoperative asymptomatic pharyngeal swallowing dysfunction predisposes for the development of symptoms of dysphagia after uvulopalatopharyngoplasty (UPPP) and uvulopalatoplasty (UPP). A total of 42 patients who snored were scheduled to undergo UPPP (n = 20) or UPP (n = 22). UPP was performed using either a CO2 laser or a conventional steel scalpel. Preoperatively and 1 year postoperatively all patients were examined videoradiographically to assess pharyngeal swallowing function. They also completed a questionnaire pre- and postoperatively concerning their snoring problems and swallowing function as well as the outcome of surgery. Preoperatively, 7 (17%) patients reported dysphagia. Pharyngeal swallowing dysfunction was demonstrated in 6/7 patients with preoperative dysphagia while pharyngeal swallowing dysfunction was evident preoperatively in 18/35 non-dysphagic patients. Of the 35 patients without preoperative dysphagia, 10 (29%/) developed dysphagia after surgery. There was no significant risk of development of postoperative dysphagia for patients with compared to patients without preoperative pharyngeal swallowing dysfunction. Only one of the seven patients with preoperative dysphagia experienced worsening of the problem. A total of 93% of the patients reported a decrease in snoring and 95% reported a decrease in daytime sleepiness. Preoperative pharyngeal swallowing dysfunction was not proven to predict the development of dysphagia after UPPP or UPP. The surgical method did not influence the frequency of postoperatively acquired dysphagia. The results do not indicate that patients with preoperative dysphagia should be excluded from treatment with UPPP or UPP.


SUN1 and SUN2 play critical but partially redundant roles in anchoring nuclei in skeletal muscle cells in mice.
How the nuclei in mammalian skeletal muscle fibers properly position themselves relative to the cell body is an interesting and important cell biology question. In the syncytial skeletal muscle cells, more than 100 nuclei are evenly distributed at the periphery of each cell, with 3-8 nuclei anchored beneath the neuromuscular junction (NMJ). Our previous studies revealed that the KASH domain-containing Syne-1/Nesprin-1 protein plays an essential role in anchoring both synaptic and nonsynaptic myonuclei in mice. SUN domain-containing proteins (SUN proteins) have been shown to interact with KASH domain-containing proteins (KASH proteins) at the nuclear envelope (NE), but their roles in nuclear positioning in mice are unknown. Here we show that the synaptic nuclear anchorage is partially perturbed in Sun1, but not in Sun2, knockout mice. Disruption of 3 or all 4 Sun1/2 wild-type alleles revealed a gene dosage effect on synaptic nuclear anchorage. The organization of nonsynaptic nuclei is disrupted in Sun1/2 double-knockout (DKO) mice as well. We further show that the localization of Syne-1 to the NE of muscle cells is disrupted in Sun1/2 DKO mice. These results clearly indicate that SUN1 and SUN2 function critically in skeletal muscle cells for Syne-1 localization at the NE, which is essential for proper myonuclear positioning.


Surname affinity in Santiago, Chile: A network-based approach that uncovers urban segregation.
Based on a geocoded registry of more than four million residents of Santiago, Chile, we build two surname-based networks that reveal the city's population structure. The first network is formed from paternal and maternal surname pairs. The second network is formed from the isonymic distances between the city's neighborhoods. These networks uncover the city's main ethnic groups and their spatial distribution. We match the networks to a socioeconomic index, and find that surnames of high socioeconomic status tend to cluster, be more diverse, and occupy a well-defined quarter of the city. The results are suggestive of a high degree of urban segregation in Santiago.


Hormonal Orchestration of Bud Dormancy Cycle in Deciduous Woody Perennials.
Woody perennials enter seasonal dormancy to avoid unfavorable environmental conditions. Plant hormones are the critical mediators regulating this complex process, which is subject to the influence of many internal and external factors. Over the last two decades, our knowledge of hormone-mediated dormancy has increased considerably, primarily due to advancements in molecular biology, omics, and bioinformatics. These advancements have enabled the elucidation of several aspects of hormonal regulation associated with bud dormancy in various deciduous tree species. Plant hormones interact with each other extensively in a context-dependent manner. The dormancy-associated MADS (DAM) transcription factors appear to enable hormones and other internal signals associated with the transition between different phases of bud dormancy. These proteins likely hold a great potential in deciphering the underlying mechanisms of dormancy initiation, maintenance, and release. In this review, a recent understanding of the roles of plant hormones, their cross talks, and their potential interactions with DAM proteins during dormancy is discussed.


Subtleties of biomineralisation revealed by manipulation of the eggshell membrane.
Biocalcification of collagen matrices with calcium phosphate and biosilicification of diatom frustules with amorphous silica are two discrete processes that have intrigued biologists and materials scientists for decades. Recent advancements in the understanding of the mechanisms involved in these two biomineralisation processes have resulted in the use of biomimetic strategies to replicate these processes separately using polyanionic, polycationic or zwitterionic analogues of extracellular matrix proteins to stabilise amorphous mineral precursor phases. To date, there is a lack of a universal model that enables the subtleties of these two apparently dissimilar biomineralisation processes to be studied together. Here, we utilise the eggshell membrane as a universal model for differential biomimetic calcification and silicification. By manipulating the eggshell membrane to render it permeable to stabilised mineral precursors, it is possible to introduce nanostructured calcium phosphate or silica into eggshell membrane fibre cores or mantles. We provide a model for infiltrating the two compartmental niches of a biopolymer membrane with different intrafibre minerals to obtain materials with potentially improved structure-property relationships.


Introduction of Carbonyl Groups into Antibodies.
Antibodies and their derivatives (scFv, Fabs, etc.) represent a unique class of biomolecules that combine selectivity with the ability to target drug delivery. Currently, one of the most promising endeavors in this field is the development of molecular diagnostic tools and antibody-based therapeutic agents, including antibody-drug conjugates (ADCs). To meet this challenge, it is imperative to advance methods for modifying antibodies. A particularly promising strategy involves the introduction of carbonyl groups into the antibody that are amenable to further modification by biorthogonal reactions, namely aliphatic, aromatic, and α-oxo aldehydes, as well as aliphatic and aryl-alkyl ketones. In this review, we summarize the preparation methods and applications of site-specific antibody conjugates that are synthesized using this approach.


Relief of pain by infusion of morphine after operation: does tolerance develop?
To see whether continuous intravenous infusion of opiates provides more effective postoperative relief of pain than conventional intramuscular injection these regimens were compared in a prospective double blind trial. Thirty patients undergoing elective cholecystectomy were allocated randomly to receive an infusion of morphine or an infusion of placebo (control group) for 24 hours. Both groups were allowed supplementary morphine boluses as requested. During the first 48 hours after operation the degree of pain was almost identical between the groups. Surprisingly, the group that was given the infusion of morphine received as much supplementary morphine as the control group during the first 24 hours and appreciably more during the 24 hours after the infusion had been withdrawn. Nausea and vomiting were more prevalent among the patients given the infusion of morphine. These results suggest that continuous infusion of morphine may be an inferior regimen to intermittent bolus administration in the relief of postoperative pain. This may be explained by the development of tolerance in patients who received the infusion of morphine.


[Comparative diagnostic value of Helicobacter pylori infection testing methods].
In 213 patients with gastric and duodenal pathology, including received surgery, comparative estimation of results of Helicobacter pylori (Hp) infection testing with invasive and noninvasive methods, were performed. Material for invasive endoscopic biopsy test (EBT), including rapid urease test (RUT) for rapid Hp identification by determination of urease activity, smear cytology and histology was extracted on endoscopy or intraoperationally. RUT was carried out with the help of URE-HP test kit. Serological test for Hp antibodies IgG and IgA class was performed by IFA using kit ELISA. 13С urea breath test (UBT) was made by determination of 13/12CO2 in breath samples on infrared spectroscope. Based on 5 different methods of Hp infection testing Hp positivity in 172 (80,8%) and Hp negativity in 41 (19,2%) from 213 examined patients was revealed. 13С-UBT revealed the highest diagnostic value (accuracy-97,5%, sensibility-97,0%, specificity-100%) in Hp infection diagnostics. In treatment efficiency control this parameters of 13С-UBT are also much high (96,7%, 90,0% and 100% respectively). In spite of high sensitivity of serological test (100%), it had comparative low specificity (71,0%) with high probability of false positive results in treated patients (antibodies titer to Hp after eradication retains for a long time). Thought, this test may be successfully used only in primary patients and in epidemiological studies. Among three methods of EBT, Hp infection detection with RUT revealed the best results (accuracy-94,8%, sensibility-95,0%, specificity-100%). Correlation of RUT and UBT results and much higher diagnostic value of UBT, necessitate RUT with histological study of stomach body mucosa to perform in patients over 45 year, with prolonged anamnesis and dangerous symptoms of disease. In Hp positive patients correlation of index DOB‰ of breath test with results of RUT was revealed. This can serve the index DOB‰ of 13С-UBT as a marker of Hp infection rate. The (accuracy, sensibility and specificity of breath test with local made 13С-urea (98,7%, 98,5% and 100% respectively) are the same as those for BT with standard 13С-urea (96,7%, 96,2% and 100% respectively). Above mentioned, 13С-UBT advantages (noninvasiveness, simplicity, rapidity, safety) and high diagnostic value exceeding same of other applied tests, give us opportunity to offer 13С-UBT as screening method of Hp infection diagnosis and a method of choise in control of Hp infection treatment efficiency.


ALKBH5 protects against stroke by reducing endoplasmic reticulum stress-dependent inflammation injury via the STAT5/PERK/EIF2α/CHOP signaling pathway in an m6A-YTHDF1-dependent manner.
Endoplasmic reticulum (ER) stress causes neuroinflammation and neuronal apoptosis during ischemic stroke progression. This study has investigated the role of ALKBH5 in ER stress during ischemic stroke progression. In vivo and in vitro models of ischemic stroke were established by middle cerebral artery occlusion (MCAO) and OGD/R treatment, respectively. Cerebral infarct size was detected using triphenyltetrazolium chloride staining (TTC), and pathological changes were examined using histological staining. The levels of inflammatory factors were analyzed using Enzyme-linked immunosorbent assay. Cell counting kit-8 assay and flow cytometry were used to measure cell viability and apoptosis, respectively. The global m6A level was detected using the commercial kit, and STAT5 mRNA m6A level was determined using methylated RNA binding protein immunoprecipitation (Me-RIP). ALKBH5, YTHDF1, and STAT5 interactions were analyzed using RIP and RNA pull-down assays. ALKBH5 was upregulated in MCAO animals and OGD/R cell models. ALKBH5 knockdown exacerbated ER stress, neuroinflammation, and neuronal apoptosis in brain tissues and neuronal cells. ALKBH5 inhibited STAT5 mRNA stability and expression in an m6A-YTHDF1-dependent manner. STAT5 promoted ER stress by activating the PERK/eIF2/CHOP signaling pathway. Furthermore, STAT5 knockdown reversed the effects of ALKBH5 knockdown on OGD/R-induced ER stress and neuroinflammation in HT22 cells. ALKBH5 knockdown exacerbated ischemic stroke by increasing ER stress-dependent neuroinflammation and neuronal apoptosis via the STAT5/PERK/EIF2α/CHOP signaling pathway in an m6A-YTHDF1-dependent manner.


Preparation of new magnetic nanocatalysts based on TiO2 and ZnO and their application in improved photocatalytic degradation of dye pollutant under visible light.
Photocatalytic degradation of methyl orange (MO) as a model of an organic pollution was accomplished with magnetic and porous TiO(2)/ZnO/Fe(3)O(4)/PANI and ZnO/Fe(3)O(4)/PANI nanocomposites under visible light irradiation. The structures of nanocomposites were characterized by various techniques including UV-Vis absorption spectroscopy, XRD, SEM, EDS, BET and TGA. Optical absorption investigations show two λ(max) at 450 and 590 nm for TiO(2)/ZnO/Fe(3)O(4)/PANI nanocomposites respectively possessing optical band gaps about 2.75 and 2.1 eV smaller than that of the neat TiO(2) and ZnO nanoparticles. Due to these optical absorptions, the nanocomposites can be considered promising candidates as visible light photocatalysts to produce more electron-hole pairs. The degradation of MO, extremely increased using polymeric photocatalysts and decolorization in the presence of visible light achieved up to 90% in less than 20 min in comparison with the neat nanoparticles (about 10%). All these advantages promise a bright future for these composites as useful photocatalysts. The degradation efficiency of MO using stable nanocomposites was still over 70% after ten times reusing. The highest decolorizing efficiencies were achieved with 0.75 g L(-1) of catalyst and 10 mg L(-1) of MO at natural pH under visible light irradiation in less than 20 min.


Concussion in Adolescents Impairs Heart Rate Response to Brief Handgrip Exercise.
Test the hypotheses that (1) concussion in adolescents impairs autonomic neural control of heart rate (HR), and (2) HR reactivity improves with symptom resolution. Observational, case-control. Nineteen concussed adolescents (8 female adolescents; age 15 ± 2 years) and 16 healthy controls (6 female adolescents, age 15 ± 2 years). All participants performed an isometric handgrip (IHG) at 30% maximum voluntary contraction lasting 30 seconds. Heart rate (electrocardiogram) and hemodynamic responses (photoplethysmographic Finometer) were recorded from 30 seconds of baseline and the last 10 seconds of handgrip. The HR response (ΔHR) at the onset of moderate-intensity IHG using a mixed 1-way analysis of variance. A group × time interaction (P < 0.005) indicated that handgrip evoked a greater ΔHR among control participants (13 ± 10 beats/min) compared with concussed (6.4 ± 6.3 beats/min; group P = 0.63; time P < 0.001; d = 0.77). These preliminary results suggest that a concussion impairs the ability to elevate HR at the exercise onset and, given the nature of the task, this could be interpreted to reflect reduced ability to withdraw cardiovagal control. Therefore, the data support the hypothesis of neural cardiac dysregulation in adolescents diagnosed with concussion. The IHG test could aid concussion diagnosis and support return-to-play decisions.


Levetiracetam Modulates Brain Metabolic Networks and Transcriptomic Signatures in the 5XFAD Mouse Model of Alzheimer's disease.
Subcritical epileptiform activity is associated with impaired cognitive function and is commonly seen in patients with Alzheimer's disease (AD). The anti-convulsant, levetiracetam (LEV), is currently being evaluated in clinical trials for its ability to reduce epileptiform activity and improve cognitive function in AD. The purpose of the current study was to apply pharmacokinetics (PK), network analysis of medical imaging, gene transcriptomics, and PK/PD modeling to a cohort of amyloidogenic mice to establish how LEV restores or drives alterations in the brain networks of mice in a dose-dependent basis using the rigorous preclinical pipeline of the MODEL-AD Preclinical Testing Core. Chronic LEV was administered to 5XFAD mice of both sexes for 3 months based on allometrically scaled clinical dose levels from PK models. Data collection and analysis consisted of a multi-modal approach utilizing 18 F-FDG PET/MRI imaging and analysis, transcriptomic analyses, and PK/PD modeling. Pharmacokinetics of LEV showed a sex and dose dependence in C max , CL/F, and AUC 0-∞ , with simulations used to estimate dose regimens. Chronic dosing at 10, 30, and 56 mg/kg, showed 18 F-FDG specific regional differences in brain uptake, and in whole brain covariance measures such as clustering coefficient, degree, network density, and connection strength (i.e. positive and negative). In addition, transcriptomic analysis via nanoString showed dose-dependent changes in gene expression in pathways consistent 18 F-FDG uptake and network changes, and PK/PD modeling showed a concentration dependence for key genes, but not for network covariance modeling. This study represents the first report detailing the relationships of metabolic covariance and transcriptomic network changes resulting from LEV administration in 5XFAD mice. Overall, our results highlight non-linear kinetics based on dose and sex, where gene expression analysis demonstrated LEV dose- and concentration-dependent changes, along with cerebral metabolism, and/or cerebral homeostatic mechanisms relevant to human AD, which aligned closely with network covariance analysis of 18 F-FDG images. Collectively, this study show cases the value of a multimodal connectomic, transcriptomic, and pharmacokinetic approach to further investigate dose dependent relationships in preclinical studies, with translational value towards informing clinical study design.


Protein stability at high temperatures.
The enzymology of hyperthermophilic micro-organisms is a growing field. As increasing numbers of novel high-temperature organisms are isolated and made available through culture collections, and, as biomass becomes more readily available, more laboratories will undoubtedly expand their research interests into this area. The prospect of totally novel enzyme systems and of new approaches to the investigation of fundamental molecular properties will continue to stimulate interest in this field. Studies of thermostable enzymes have already provided valuable data on the relationships between protein stability and activity. The subtle molecular mechanisms which have evolved to stabilize these proteins provide the clues needed for the intelligent design of stabilized mesophilic enzymes, an important target where a combination of high activity at 'low' temperatures and resistance to denaturation is required. The current role of hyperthermophilic enzymes in biotechnology is relatively minor, despite these enzymes having a high 'profile'. While early over-enthusiastic predictions that these enzymes would revolutionize biotechnology should be disregarded, it can reasonably be assumed that, where functional and economic criteria are suitable, thermophilic enzymes will be readily incorporated into current and future biotechnology.


[Exploring Molecular Targets for Epilepsy Treatment from the Perspective of Neuronal Homeostasis].
Brain function is controlled by the balance between the excitatory and inhibitory systems. If this balance is disrupted and the excitatory system dominates, convulsions or epileptic seizures are induced. Neuronal hyperexcitability in the brain leads to marked changes in the function of the neurons, which adversely affect the stability of the neural network. Many of the currently used antiepileptic drugs are symptomatic treatments that suppress the electrical hyperexcitability of the cerebrum. Although patients with epilepsy should continuously take antiepileptic drugs to control their seizures, approximately 20% of patients are drug resistant. The brain has the ability to control neuronal functions within acceptable limits while it maintains the amount of synaptic inputs that form the basis of information accumulation. Neuronal self-regulation is known as homeostatic scaling by which the intensity of all excitatory synapses is suppressed when neuronal excitability is increased. However, the molecular mechanisms of homeostatic scaling and their pathophysiological significance in vivo remain unclear. Repeated treatment with a subconvulsive dosage of pentylenetetrazol (PTZ), a γ-aminobutyric acid (GABA)A receptor antagonist, is known to induce kindling in mice, which is a common animal model used to study epilepsy. We found that PTZ-induced kindling was potentiated in mice deficient in the transcription factor neuronal PAS domain protein 4 (Npas4), the expression of which is immediately induced in response to neuronal activity. At this symposium, we will discuss the possibility of Npas4 as a novel target molecule for epilepsy treatment.


Water imbibition in the rat prostate during castration-induced regression.
The present study was conducted to examine changes in water concentration in the ventral prostate of adult rats at different intervals following castration. The prostatic dry weight was obtained by drying the fresh prostate at 70 degrees C for at least 110 hr and the prostatic water content was calculated from its wet and dry weight. Under normal conditions, the prostatic water concentration ranges from 81.9 to 82.7% in uncastrated rats. The prostatic water concentration started to increase at 8 hr postcastration (83.1%) but this increase was not statistically significant. By 16 hr postcastration, the prostatic water concentration (83.8%) became significantly higher than that of the uncastrated animals. In rats of day 1 to day 10 postcastration, the prostate contained significantly more water (84.4%-84.7%) than those in uncastrated animals. By day 21 postcastration, the prostatic water concentration (81.5%) was not significantly different from that of uncastrated rats. Unlike the prostate, the skeletal muscle did not significantly change the water concentration following castration. The 51Cr-EDTA space in the prostate was not significantly different in rats before and after castration. These results indicate that water imbibition in the rat prostate is associated with an active period of prostatic regression and that the change in the 51Cr-EDTA space during prostatic regression is not the major cause of water imbibition in the tissue. Therefore, the present findings suggest that castration-induced water imbibition in the rat prostate is caused by an increase in the intracellular water space.


Induction of puberty with human chorionic gonadotropin (hCG) followed by reversal of hypogonadotropic hypogonadism in Kallmann syndrome.
Kallmann syndrome (KS) is a rare, congenital disorder combining hypogonadotropic hypogonadism (HH) due to GnRH-deficiency with anosmia. Traditionally thought to require lifelong therapy it turns out to be a reversible condition in some patients. We present a case of a 22-year old man with absent puberty due to KS, in whom genetic testing revealed heterozygosity for a mutation in the PROK2 gene. Pubertal development and virilisation was achieved by using human chorionic gonadotropin (hCG) injections followed by testosterone replacement. During the follow-up we observed reversal of hypogonadism allowing discontinuation of testosterone treatment. Normalisation of testicular volume as well as gonadotropin and inhibin B levels through a 2-year postreversal period was seen. Treatment with hCG is effective in inducing pubertal development and may have advantage over testosterone replacement due to a potential of gonadal maturation. A regular assessment of testicular volume and biochemical surveillance including measuring of serum inhibin B and gonadotropins are necessary for a timely detection of reversal of GnRH deficiency.


Formative study to develop the Eban treatment and comparison interventions for couples.
To describe formative and pilot-testing research that generated themes and procedures, curricula, and critical measures for a randomized clinical trial testing a Risk Reduction Intervention for HIV-serodiscordant African American couples (Project Eban). This article describes the themes that emerged from discussions with African American serodiscordant couples about HIV-related risks from focus groups with 11 couples and pilot study results with 32 couples across 4 sites. In step 1, focus groups examined the need for a Risk Reduction Intervention for HIV-serodiscordant African American couples and confirmed 4 themes that formed the basis for the intervention curriculum and study format. In step 2, a pilot study refined the clinical trial procedures for this population and tested critical measures and selected portions of the curriculum for both the treatment and the comparison interventions. Based on these findings, stigma and psychological distress, barriers to condom use, insufficient support from community and service organizations, and the lack of skills that emphasize individual and relationship protection were ultimately integrated into the Risk Reduction Intervention. Pilot study findings highlighted the importance of examining gender and ethnicity in HIV-impacted couples along with factors that heightened HIV-related risk behaviors that affect couples' skills and psychological adjustment. The goal was to ensure that a skill-based, culturally congruent, relationship-centered intervention could be understood and of interest to couples. Future analyses in the main trial will be discussed.


Chemotherapy-Induced IL34 Enhances Immunosuppression by Tumor-Associated Macrophages and Mediates Survival of Chemoresistant Lung Cancer Cells.
The ability of tumor cells to escape immune destruction and their acquired resistance to chemotherapy are major obstacles to effective cancer therapy. Although immune checkpoint therapies such as anti-PD-1 address these issues in part, clinical responses remain limited to a subpopulation of patients. In this report, we identified IL34 produced by cancer cells as a driver of chemoresistance. In particular, we found that IL34 modulated the functions of tumor-associated macrophages to enhance local immunosuppression and to promote the survival of chemoresistant cancer cells by activating AKT signaling. Targeting IL34 in chemoresistant tumors resulted in a remarkable inhibition of tumor growth when accompanied with chemotherapy. Our results define a pathogenic role for IL34 in mediating immunosuppression and chemoresistance and identify it as a tractable target for anticancer therapy. Cancer Res; 76(20); 6030-42. ©2016 AACR.


Clonidine and left ventricular function in ten patients with essential hypertension.
Left ventricular mass and function were evaluated by M-mode echocardiography in 10 patients with essential hypertension, before and after 16 weeks of treatment with clonidine. Plasma renin activity (PRA), serum and 24-h urinary Na and K were also determined. The dose range of clonidine was 0.15-0.45 mg/day. Systolic and diastolic blood pressure decreased significantly since the second week of treatment (p less than 0.0001). There was a decrease in calculated total peripheral resistance (35.3 +/- 11.08 to 30.91 +/- 6.6 RU p less than 0.01), end meridional (72.9 +/- 23.6 to 58.5 +/- 22 dynes/cm2 X 10(3), p less than 0.001) and peak systolic wall stress (453.5 +/- 140 to 378 +/- 97 dynes/cm2 X 10(3), p less than 0.01), wall tension (333 +/- 36 to 286.6 +/- 42, p less than 0.01). Left ventricular mass index decreased in six patients (83.7 +/- 20.4 to 70.7 +/- 12.4 g/m2, p less than 0.001). Decrease in left ventricular mass index was related to decrease in wall tension (r = 0.70 p less than 0.01) and septal thickness in diastole (r = 0.71, p less than 0.01). There was a decrease in "double product" (10,934 +/- 1,975 to 8,705 +/- 1,739, p less than 0.01), PRA (3.74 +/- 2.51 to 2.51 +/- 1.44 ng/ml/h p less than 0.01) with increase in 24-h urinary Na (114.9 +/- 37 to 155.3 +/- 55.8 mEq/24-h urine, p less than 0.001). Cardiac index, left ventricular ejection fraction and fractional shortening increased without significance (p = NS).(ABSTRACT TRUNCATED AT 250 WORDS)


Data-free prior model for upper body pose estimation and tracking.
Video based human body pose estimation seeks to estimate the human body pose from an image or a video sequence, which captures a person exhibiting some activities. To handle noise and occlusion, a pose prior model is often constructed and is subsequently combined with the pose estimated from the image data to achieve a more robust body pose tracking. Various body prior models have been proposed. Most of them are data-driven, typically learned from 3D motion capture data. In addition to being expensive and time-consuming to collect, these data-based prior models cannot generalize well to activities and subjects not present in the motion capture data. To alleviate this problem, we propose to learn the prior model from anatomic, biomechanics, and physical constraints, rather than from the motion capture data. For this, we propose methods that can effectively capture different types of constraints and systematically encode them into the prior model. Experiments on benchmark data sets show the proposed prior model, compared with data-based prior models, achieves comparable performance for body motions that are present in the training data. It, however, significantly outperforms the data-based prior models in generalization to different body motions and to different subjects.


Growth characteristics of poliomyelitis virus in HeLa cell cultures; lack of parallelism in cellular injury and virus increase.
A detailed study of the cytological changes which are induced in HeLa cells by the Saukett strain of Type III poliomyelitis virus has been made. The observations were of cultures in which a single sequence of infection was induced. The cytological changes were examined in relation to the growth curve of the virus in the same type of culture. This curve showed a latent period of 4 to 5 hours, followed by a gradual release of virus over an interval of 6 to 7 hours. Changes in the staining character of the cells occurred before the major portion of the viral yield appeared. The infected cells exhibited a striking cytopathology with increased basophilia, nuclear pyknosis, and basophilic cytoplasmic granules. Individual cells showed characteristic differences in the rate at which the cytopathology progresses. The multiplication of the virus in HeLa cells was inhibited by fluorophenylalanine. The inhibitory effectiveness of the antimetabolite was related to the age of the infection. It apparently inhibits only an early stage of viral development. The inhibition is completely reversed by phenylalanine if the amino acid is added within 6 hours, not later, after the induction of virostasis. The data are interpreted in terms of the rate at which the ability of the infected cell to support viral synthesis was lost. Flurophenylalanine also inhibited the multiplication of HeLa cells; however, the effect upon the uninfected cell was reversible after 3 days, as indicated by viability after such treatment. While the fluoro derivative completely inhibited viral multiplication, it did not prevent the cytopathogenic effect of the virus. In the presence of fluorophenylalanine, the disintegration of an infected cell proceeded at what appeared to be the ordinary rate, without any increase of the infectious agent. Experimentally the processes leading to viral increase and to cellular injury have been shown to possess a significant degree of autonomy.


Therapeutic apheresis services provided by a community blood bank.
Therapeutic apheresis services can be provided from a community-based blood center. The Oklahoma Blood Institute uses a mobile approach, primarily servicing 10 major hospitals in the Oklahoma City metropolitan area within a 15 to 20 mile radius. Apheresis staff and equipment are available 24 hours per day, 7 days per week in the event of patient referral requiring an emergency therapeutic apheresis procedure. This model requires close communication between the therapeutic apheresis medical director and the referring clinician. Telephone and facsimile transmissions of information are used to facilitate the transfer of information to all persons/facilities involved in providing treatment. This approach has proven effective in providing the required procedures for a referral population base of several million people spread over a fairly large geographic area.


Purified and unpurified sodium channels from eel electroplax in planar lipid bilayers.
Highly purified sodium channel protein from the electric eel, Electrophorus electricus, was reconstituted into liposomes and incorporated into planar bilayers made from neutral phospholipids dissolved in decane. The purest sodium channel preparations consisted of only the large, 260-kD tetrodotoxin (TTX)-binding polypeptide. For all preparations, batrachotoxin (BTX) induced long-lived single-channel currents (25 pS at 500 mM NaCl) that showed voltage-dependent activation and were blocked by TTX. This block was also voltage dependent, with negative potentials increasing block. The permeability ratios were 4.7 for Na+:K+ and 1.6 for Na+:Li+. The midpoint for steady state activation occurred around -70 mV and did not shift significantly when the NaCl concentration was increased from 50 to 1,000 mM. Veratridine-induced single-channel currents were about half the size of those activated by BTX. Unpurified, nonsolubilized sodium channels from E. electricus membrane fragments were also incorporated into planar bilayers. There were no detectable differences in the characteristics of unpurified and purified sodium channels, although membrane stability was considerably higher when purified material was used. Thus, in the eel, the large, 260-kD polypeptide alone is sufficient to demonstrate single-channel activity like that observed for mammalian sodium channel preparations in which smaller subunits have been found.


Enhanced chondrogenesis of mesenchymal stem cells in collagen mimetic peptide-mediated microenvironment.
A new type of synthetic hydrogel scaffold that mimics certain aspects of structure and function of natural extracellular matrix (ECM) has been developed. We previously reported the conjugation of collagen mimetic peptide (CMP) to poly(ethylene oxide) diacrylate (PEODA) to create a polymer-peptide hybrid scaffold for a suitable cell microenvironment. In this study, we showed that the CMP-mediated microenvironment enhances the chondrogenic differentiation of mesenchymal stem cells (MSCs). MSCs were harvested and photo-encapsulated in CMP-conjugated PEODA (CMP/PEODA). After 3 weeks, the histological and biochemical analysis of the CMP/PEODA gel revealed twice as much glycosaminoglycan and collagen contents as in control PEODA hydrogels. Moreover, MSCs cultured in CMP/PEODA hydrogel exhibited a lower level of hypertrophic markers, core binding factor alpha 1, and type X collagen than MSCs in PEODA hydrogel as revealed by gene expression and immunohistochemisty. These results indicate that CMP/PEODA hydrogel provides a favorable microenvironment for encapsulated MSCs and regulates their downstream chondrogenic differentiation.


Early glycoprotein IIb-IIIa inhibitors in primary angioplasty (EGYPT) cooperation: an individual patient data meta-analysis.
Even though time-to-treatment has been shown to be a determinant of mortality in primary angioplasty, the potential benefits from early pharmacological reperfusion by glycoprotein (Gp) IIb-IIIa inhibitors are still unclear. The aim of this meta-analysis was to combine individual data from all randomised trials conducted on facilitated primary angioplasty by the use of early Gp IIb-IIIa inhibitors. The literature was scanned by formal searches of electronic databases (MEDLINE, EMBASE) from January 1990 to October 2007. All randomised trials on facilitation by the early administration of Gp IIb-IIIa inhibitors in ST-segment elevation myocardial infarction (STEMI) were examined. No language restrictions were enforced. Individual patient data were obtained from 11 out of 13 trials, including 1662 patients (840 patients (50.5%) randomly assigned to early and 822 patients (49.5%) to late Gp IIb-IIIa inhibitor administration). Preprocedural Thrombolysis in Myocardial Infarction Study (TIMI) grade 3 flow was more frequent with early Gp IIb-IIIa inhibitors. Postprocedural TIMI 3 flow and myocardial blush grade 3 were higher with early Gp IIb-IIIa inhibitors but did not reach statistical significance except for abciximab, whereas the rate of complete ST-segment resolution was significantly higher with early Gp IIb-IIIa inhibitors. Mortality was not significantly different between groups, although early abciximab demonstrated improved survival compared with late administration, even after adjustment for clinical and angiographic confounding factors. This meta-analysis shows that pharmacological facilitation with the early administration of Gp IIb-IIIa inhibitors in patients undergoing primary angioplasty for STEMI is associated with significant benefits in terms of preprocedural epicardial recanalisation and ST-segment resolution, which translated into non-significant mortality benefits except for abciximab.


Impact of diisobutyl phthalate and other PPAR agonists on steroidogenesis and plasma insulin and leptin levels in fetal rats.
Endocrine disrupting chemicals can induce malformations and impairment of reproductive function in experimental animals and may have similar effects in humans. Recently, the environmental obesogen hypothesis was proposed, suggesting that environmental chemicals contribute to the development of obesity and insulin resistance. These effects could be related to chemical interaction with nuclear receptors such as the peroxisome proliferator activated receptors (PPARs). As several testosterone-reducing drugs are PPAR activators, we aimed to examine whether four PPAR agonists were able to affect fetal testosterone production and masculinization of rats. Additionally, we wished to examine whether these chemicals affected fetal plasma levels of insulin and leptin, which play important roles in the developmental programming of the metabolic system. Pregnant Wistar rats were exposed from gestation day (GD) 7-21 to diisobutyl phthalate (DiBP), butylparaben, perfluorooctanoate, or rosiglitazone (600, 100, 20, or 1 mg/kg bw/day, respectively). Endocrine endpoints were studied in offspring at GD 19 or 21. DiBP, butylparaben and rosiglitazone reduced plasma leptin levels in male and female offspring. DiBP and rosiglitazone additionally reduced fetal plasma insulin levels. In males, DiBP reduced anogenital distance, testosterone production and testicular expression of Insl-3 and genes related to steroidogenesis. PPARalpha mRNA levels were reduced by DiBP at GD 19 in testis and liver. In females, DiBP increased anogenital distance and increased ovarian aromatase mRNA levels. This study reveals new targets for phthalates and parabens in fetal male and female rats and contributes to the increasing concern about adverse effects of human exposure to these compounds.


Does the ingestion of alcohol influence the development of arteriosclerosis in fowls?
The effect of alcohol ingestion upon the development of naturally occurring and stilbestrol-induced arteriosclerosis was studied in the domestic fowl. In two experiments, a 15 per cent ethyl alcohol solution was used for drinking purposes, and in a third experiment wine containing 12 per cent of it was administered. The caloric intake of both food and alcohol was carefully controlled by pair-feeding, a glucose solution being used for drinking purposes to equalize the caloric intakes of the control birds with those of the alcohol-treated birds. A total of 224 cockrels was studied, and the period of observation for each experiment lasted 12 months. The degree of arteriosclerosis in the thoracic and abdominal aortas was determined in the gross and microscopically, and the degree of lipide infiltration was determined microscopically. The gross grading of arteriosclerotic lesions agreed closely with the microscopic analysis of intimal thickening and lipide infiltration of the arterial wall. The gross gradings were subjected to a critical statistical analysis which allowed precise statements to be made on the probability that a real effect would be overlooked. This analysis yielded no evidence that alcohol or the wine used had affected the degree of gross arteriosclerosis or lipide infiltration.


A Novel ALAS2 Missense Mutation in Two Brothers With Iron Overload and Associated Alterations in Serum Hepcidin/Erythroferrone Levels.
Iron loading anemias are characterized by ineffective erythropoiesis and iron overload. The prototype is non-transfusion dependent ß-thalassemia (NTDT), with other entities including congenital sideroblastic anemias, congenital dyserythropoietic anemias, some hemolytic anemias, and myelodysplastic syndromes. Differential diagnosis of iron loading anemias may be challenging due to heterogeneous genotype and phenotype. Notwithstanding the recent advances in linking ineffective erythropoiesis to iron overload, many pathophysiologic aspects are still unclear. Moreover, measurement of hepcidin and erythroferrone (ERFE), two key molecules in iron homeostasis and erythropoiesis, is scarcely used in clinical practice and of uncertain utility. Here, we describe a comprehensive diagnostic approach, including next-generation sequencing (NGS), in silico modeling, and measurement of hepcidin and erythroferrone (ERFE), in two brothers eventually diagnosed as X-linked sideroblastic anemia (XLSA). A novel pathogenic ALAS2 missense mutation (c.1382T>A, p.Leu461His) is described. Hyperferritinemia with high hepcidin-25 levels (but decreased hepcidin:ferritin ratio) and mild-to-moderate iron overload were detected in both patients. ERFE levels were markedly elevated in both patients, especially in the proband, who had a more expressed phenotype. Our study illustrates how new technologies, such as NGS, in silico modeling, and measurement of serum hepcidin-25 and ERFE, may help in diagnosing and studying iron loading anemias. Further studies on the hepcidin-25/ERFE axis in additional patients with XLSA and other iron loading anemias may help in establishing its usefulness in differential diagnosis, and it may also aid our understanding of the pathophysiology of these genetically and phenotypically heterogeneous entities.


Organizational requirements of the SaeR binding sites for a functional P1 promoter of the sae operon in Staphylococcus aureus.
In Staphylococcus aureus, the SaeRS two-component system controls the expression of multiple virulence factors. Of the two promoters in the sae operon, P1 is autoinduced and has two binding sites for the response regulator SaeR. In this study, we examined the organizational requirements of the SaeR binding sites in P1 for transcription activation. Mutational studies showed that both binding sites are essential for binding to phosphorylated SaeR (P-SaeR) and transcription activation. When the 21-bp distance between the centers of the two SaeR binding sites was altered to 26 bp, 31 bp, 36 bp, or 41 bp, only the 31-bp mutant retained approximately 40% of the original promoter activity. When the -1-bp spacing (i.e.,1-bp overlap) between the primary SaeR binding site and the -35 promoter region was altered, all mutant P1 promoters failed to initiate transcription; however, when the first nucleotide of the -35 region was changed from A to T, the mutants with 0-bp or 22-bp spacing showed detectable promoter activity. Although P-SaeR was essential for the binding of RNA polymerase to P1, it was not essential for the binding of the enzyme to the alpha-hemolysin promoter. When the nonoptimal spacing between promoter elements in P1 or the coagulase promoter was altered to the optimal spacing of 17 bp, both promoters failed to initiate transcription. These results suggest that SaeR binding sites are under rather strict organizational restrictions and provide clues for understanding the molecular mechanism of sae-mediated transcription activation.


A role for the endogenous opioid beta-endorphin in energy homeostasis.
Proopiomelanocortin (POMC) neurons in the hypothalamus are direct targets of the adipostatic hormone leptin and contribute to energy homeostasis by integrating peripheral and central information. The melanocortin and beta-endorphin neuropeptides are processed from POMC and putatively coreleased at axon terminals. Melanocortins have been shown by a combination of pharmacological and genetic methods to have inhibitory effects on appetite and body weight. In contrast, pharmacological studies have generally indicated that opioids stimulate food intake. Here we report that male mice engineered to selectively lack beta-endorphin, but that retained normal melanocortin signaling, were hyperphagic and obese. Furthermore, beta-endorphin mutant and wild-type mice had identical orexigenic responses to exogenous opioids and identical anorectic responses to the nonselective opioid antagonist naloxone, implicating an alternative endogenous opioid tone to beta-endorphin that physiologically stimulates feeding. These genetic data indicate that beta-endorphin is required for normal regulation of feeding, but, in contrast to earlier reports suggesting opposing actions of beta-endorphin and melanocortins on appetite, our results suggest a more complementary interaction between the endogenously released POMC-derived peptides in the regulation of energy homeostasis.


[Formulation of the composition of a polymyxin ointment using a mathematical experimental design].
Mathematical design of the experiment permitted detection of the action of independent factors of the component composition on antibacterial activity and rheological properties of polymyxin ointment. Optimal compositions of polymyxin ointment based on polyethylene oxide emulsion were scientifically grounded and recommended for further experimental investigation on animals.


The Belmont Report and Innovative Clinical Research.
A central pillar of the Belmont Report is that a bright line must be drawn between medical practice and biomedical research. That line may have been brighter 50 years ago. Today, the typical physician is likely to work for a corporation or health system that styles itself as a learning health system. Such systems increasingly emphasize the (research-like) use of data to measure quality, encourage efficiency, ensure safety, and guide a standardized approach to clinical care. While these activities are not considered research, they pose many of the same risks or conflicts of loyalty. In research, the doctor's fiduciary loyalty to the patient is compromised by a loyalty to the scientific process. In learning health systems, the doctor's loyalty is compromised by loyalty to the system and its metrics. In this world, it is not clear that research-as conceptualized by the Belmont Report, codified in the Common Rule, and overseen by IRBs-is a uniquely risky activity deserving of such uniquely strict oversight. Perhaps, instead, the divided loyalties and conflicts of interest faced by everyday clinicians working in learning health systems demand a protective framework similar to the one that we now have for the activities that we designate as "research." This article compares the risks of the various activities that might be called "research" and suggests a unified system of oversight for all of them.