Folding and binding integrity of variants of a prototype ligand-binding module from the LDL receptor possessing multiple alanine substitutions.
The LA repeats that comprise the ligand-binding domain of the LDL receptor are among the most common autonomously structured extracellular modules found in the nonredundant protein sequence database. Here, we investigate the information content of the amino acid sequence of a typical LA module by constructing sequences with alanine residues at nonconserved positions in the module. Starting with the sequence of the fifth ligand-binding repeat of the LDL receptor (LA5), we created generic LA modules with alanine substitutions of nonconserved residues in only the N-terminal lobe, only the C-terminal lobe, and throughout both lobes of the module. LA variants with alanine residues at as many as 18 of 37 positions fold to a preferred disulfide isomer in the presence of calcium. Indeed, the six cysteines, the C-terminal calcium coordinating residues, two hydrophobic residues involved in packing, two glycines, and five other residues that form side chain-intramodule hydrogen bonds are alone sufficient to specify the fold of an LA module when alanine residues are present at all other positions. The LA variants with multiple alanines in either the N- or C-terminal lobe were then exploited to identify residues of LA5 that contribute to the binding of apoE-containing ligands in LDL receptor-derived "minireceptors", implicating nonconserved residues of the N-terminal lobe of LA5 in recognition of apoE-DMPC. Our library of LA modules with multiple alanine substitutions should be generally useful for probing the roles of nonconserved side chains in ligand recognition by proteins of the LDL receptor family.


Negative feedback confers mutational robustness in yeast transcription factor regulation.
Organismal fitness depends on the ability of gene networks to function robustly in the face of environmental and genetic perturbations. Understanding the mechanisms of this stability is one of the key aims of modern systems biology. Dissecting the basis of robustness to mutation has proven a particular challenge, with most experimental models relying on artificial DNA sequence variants engineered in the laboratory. In this work, we hypothesized that negative regulatory feedback could stabilize gene expression against the disruptions that arise from natural genetic variation. We screened yeast transcription factors for feedback and used the results to establish ROX1 (Repressor of hypOXia) as a model system for the study of feedback in circuit behaviors and its impact across genetically heterogeneous populations. Mutagenesis experiments revealed the mechanism of Rox1 as a direct transcriptional repressor at its own gene, enabling a regulatory program of rapid induction during environmental change that reached a plateau of moderate steady-state expression. Additionally, in a given environmental condition, Rox1 levels varied widely across genetically distinct strains; the ROX1 feedback loop regulated this variation, in that the range of expression levels across genetic backgrounds showed greater spread in ROX1 feedback mutants than among strains with the ROX1 feedback loop intact. Our findings indicate that the ROX1 feedback circuit is tuned to respond to perturbations arising from natural genetic variation in addition to its role in induction behavior. We suggest that regulatory feedback may be an important element of the network architectures that confer mutational robustness across biology.


Pulmonary endarterectomy is effective and safe in patients with haemoglobinopathies and abnormal red blood cells: the Papworth experience.
Patients with haemoglobinopathies and congenital haemolytic anaemia constitute a unique population more predisposed to developing chronic thromboembolic pulmonary hypertension (CTEPH). Although pulmonary endarterectomy (PEA) is accepted as the best treatment for CTEPH, PEA in these patients poses significant practical challenges. Apart from a few case reports, the results of PEA in this patient population have not been previously reported. The aim of this study was to review the outcome of PEA in this patient population. We performed a retrospective analysis, from our dedicated CTEPH database, of all patients who underwent PEA surgery and had abnormal haemoglobin or congenital haemolytic anaemia. We reviewed diagnosis, exchange transfusions on cardiopulmonary bypass, preoperative and postoperative pulmonary haemodynamic and functional data and outcomes for this group. Paired data analysis was performed by Student's t-test; P < 0.05 was statistically significant. Between the start of our PEA programme in 1997 and April 2015, we performed PEA in 19 patients with haemoglobinopathy or congenital haemolytic anaemia. The mean age was 52 ± 15 years. There were 9 patients with sickle cell trait, 2 with coexisting alpha+ thalassaemia trait, 2 patients with HbSC disease, 2 patients with beta-thalassaemia major, 3 patients with hereditary spherocytosis, 2 patients with stomatocytosis (one with the cryohydrocytosis subtype) and 1 patient with HbC trait. In the 9 HbAS patients, the mean HbS% was 31.9 ± 6%, and in the HbSC patients, the mean HbS% was 46.5 ± 1.3% preoperatively. To reduce this HbS to ≤20%, for safe PEA with deep hypothermic circulatory arrest, we used exchange blood transfusion. Immediately postoperatively, there was a significant improvement in pulmonary vascular resistance (938 ± 462 to 260 ± 167 dyne s cm(-5); P < 0.0001). One patient died 81 days following surgery; 18 patients are alive at a median follow-up of 3.4 ± 3 years. Six months postoperatively, the patients showed significant improvement in New York Heart Association status (P < 0.0001), and in 6-min walk distance from 251 ± 111 to 399 ± 69 m (P < 0.0001). Results of PEA in this complex patient group were satisfactory. Expert haematological advice is important and exchange blood transfusions may be necessary. The presence of abnormal haemoglobin does not contra-indicate PEA surgery.


Complete genome sequence of Mahella australiensis type strain (50-1 BON).
Mahella australiensis Bonilla Salinas et al. 2004 is the type species of the genus Mahella, which belongs to the family Thermoanaerobacteraceae. The species is of interest because it differs from other known anaerobic spore-forming bacteria in its G+C content, and in certain phenotypic traits, such as carbon source utilization and relationship to temperature. Moreover, it has been discussed that this species might be an indigenous member of petroleum and oil reservoirs. This is the first completed genome sequence of a member of the genus Mahella and the ninth completed type strain genome sequence from the family Thermoanaerobacteraceae. The 3,135,972 bp long genome with its 2,974 protein-coding and 59 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.


Inflammatory Processes Associated with Canine Intervertebral Disc Herniation.
Intervertebral disc herniation (IVDH) is an important pathology in humans and also in dogs. While the molecular disease mechanisms are well investigated in humans, little is known about the inflammatory mediators in naturally occurring canine IVDH. The objective of this study was to investigate whether the involved proinflammatory cytokines in human IVDH are also key cytokines in canine IVDH and thus to elucidate the suitability of the dog as a model for human trials. 59 samples from 25 dogs with surgically confirmed thoracolumbar IVDH were collected and classified in three subgroups: herniated (H), affected non-herniated (NH) disc, and adjacent non-affected (NA) disc. Discs from 11 healthy dogs acted as controls (C). Samples were analyzed for IL-1, IL-6, IL-8, and TNF-α expression (qPCR/ELISA) as well as cell infiltration and activation of the MAP kinase pathways (immunohistochemistry). Gene and protein expression of all key cytokines could be detected in IVDH affected dogs. Canine IVDH was significantly associated with a higher gene expression of IL-6 (H > C, NH > C) and TNF-α (H > C, NH > C, NA > C) and a significant down-regulation of IL-1β (H < C). Dogs with spontaneous pain had significantly higher IL-6 mRNA compared to those with pain arising only upon palpation. An inter-donor comparison (H and HN relative to NA) revealed a significant increase of IL-6 gene expression (H > NA, NH > NA). IL-8 (H > C, NA > C) and TNF-α (NH > C) protein levels were significantly increased in diseased dogs while inversely, IL-6 protein levels were significantly higher in patients with better clinical outcome. Aside from resident IVD cells, mostly monocytes and macrophages were found in extruded material, with concomitant activation of extracellular signal-regulated kinase p38 in the majority of samples. Dogs with spontaneous IVDH might provide a useful model for human disc diseases. Although the expression of key cytokines found in human IVDH was also demonstrated in canine tissue, the inflammatory mechanisms accompanying canine IVDH diverges partially from humans, which will require further investigations in the future. In dogs, IL-6 seems to play an important pathological role and may represent a new potential therapeutic target for canine patients.


Europium(III) complexes containing organosilyldipyridine ligands grafted on silica nanoparticles.
This work focuses on the grafting of transition metal complexes on silica surface nanoparticles. Nanoscale silica particles in aqueous sols are used as starting silicated materials. We have undertaken the synthesis of europium(III) complexes containing organosilyldipyridine ligands, (EtO)3Si(CH2)3NHCH2-bipy (1) and (EtO)(CH3)2Si(CH2) 3NHCH 2-bipy (2), in view of a direct grafting reaction on silica nanoparticles. Reaction of one molar equivalent of 1 and 2 with Eu(tmhd)3 (tmhd= 2,2,6,6-tetramethyl-3,5-heptanedionato), as precursor, leads to octacoordinated silylated europium(III) complexes [Eu(tmhd)3(1)] (3) and [Eu(tmhd)3(2)] (4) as white solids in 34-54% yields. Europium complexes were characterized by elemental analysis, mass spectrometry, FT-IR, UV, and luminescence spectroscopies. These new complexes are reacting in a 1:10 (v/v) water and ethanol mixture with silica nanoparticles colloidal sol. Elemental analysis and thermogravimetric data indicated grafting ratios of 0.41 and 0.26 mmol of europium(III) complexes per gram of silica. Functionalized silica nanoparticles were characterized by DRIFT spectroscopy and TEM microscopy. The first analysis shows that the chemical integrity of the complexes is retained on the silica surface together with the size and the monodispersity of the nanoscale particles. As expected for europium(III) complexes, luminescence is observed under UV irradiation. Emission and excitation spectra indicate that the metal coordination environment is not modified on the silica surface. Moreover, the sharpness of the luminescence bands and the strong antenna effect are maintained when complexes are covalently bonded to silica. New luminescent europium(III) complexes grafted on silica nanoparticles are therefore obtained from our approach.


Reflections on abortion rights: From policy to medicine.
On June 24, 2022, the US Supreme Court overturned Roe v. Wade, which marked a further restriction on women's abortion rights in the US. It has sparked a wide range of societal reactions around the world. Women in different countries enjoy diverse abortion rights due to conditions in their respective nations and cultures. Abortion protects women's rights to a certain extent, and especially in the event of unintended pregnancy. An inappropriate abortion ban will affect women's health and lives and all aspects of medicine, including the lives of doctors, patient access, and the development of medical technology. This review provides a gynecologist's perspective on the impact of abortion policies on women's health and the medical system. This review also attempts to determine the reason for the government's abortion ban.


Periodic Heat Stress Licenses EMSC Differentiation into Osteoblasts via YAP Signaling Pathway Activation.
The repair and regeneration of large bone defects represent highly challenging tasks in bone tissue engineering. Although recent studies have shown that osteogenesis is stimulated by periodic heat stress, the thermal regulation of osteogenic differentiation in ectomesenchymal stem cells (EMSCs) is not well studied. In this study, the direct effects of periodic heat stress on the differentiation of EMSCs into osteoblasts were investigated. EMSCs derived from rat nasal respiratory mucosa were seeded onto culture plates, followed by 1 h of heat stress at 41°C every 7 days during osteogenic differentiation. Based on the results of the present study, periodic heating increases alkaline phosphatase (ALP) activity, upregulates osteogenic-related proteins, and promotes EMSC mineralization. In particular, increased YAP nuclear translocation and YAP knockdown inhibited osteogenic differentiation induced by heat stress. Furthermore, the expression and activity of transglutaminase 2 (TG2) were significantly increased after YAP nuclear translocation. Together, these results indicate that YAP plays a key role in regulating cellular proteostasis under stressful cellular conditions by modulating the TG2 response.


[Treatment of fresh muscle injury].
A prospective double-blind randomized study comparing the results of a protein-free-haemodialisate (Actovegin) with placebo was performed in 103 patients. There has been 68 patients in the haemodialysate group. In acute muscle injuries we find a dent in the muscle and a loss of function. There has been three injections in the injured muscle every three to four days. The follow up examination occurred at least three months after the beginning of the study. Full sports activity was reached in the haemodialysate group after 5.5 weeks, in the non-haemodialysate group after 8.3 weeks. There has been a significant difference in the time of rehabilitation between the shank and the thigh muscles. The results show a high statistic significance.


Anterior abdominal wall abscess with epididymo-orchitis: an unusual presentation of acute pancreatitis.
Pancreatitis indicates inflammation of the pancreas. Clinically acute pancreatitis typically presents as upper abdominal pain mostly in epigastric region, nausea, vomiting and elevated levels of amylase and lipase. Depending upon severity of acute pancreatitis patient may presents with minimal symptoms to more severe signs of acute abdomen like generalized guarding and rigidity. Inspite of absence of disease-specific signs and symptoms for acute pancreatitis, diagnosis is usually not difficult using a combination of clinical, laboratory and radiological findings. Sometimes pancreatitis may presents atypically, which may be misleading in the management especially when typical presentation of pancreatitis as described above is absent. We have described a case of pancreatitis where patient presented with anterior abdominal wall abscess with epididymo-orchitis because of tracking of pancreatic fluid into the retroperitoneum till scrotum. Patients presentation may be different depending upon complication occurred during the course of pancreatitis. After reviewing the literature we found very few cases in which you may not get a clue to diagnose pancreatitis because of atypical presentation. In the described case, patient managed conservatively with percutaneous drainage of the abscess by pigtail catheter placement and scrotal support for epididymoorchitis. This avoided unnecessary exploration in above patient.


Marijuana in the management of amyotrophic lateral sclerosis.
Marijuana has been proposed as treatment for a widening spectrum of medical conditions. Marijuana is a substance with many properties that may be applicable to the management of amyotrophic lateral sclerosis (ALS). These include analgesia, muscle relaxation, bronchodilation, saliva reduction, appetite stimulation, and sleep induction. In addition, marijuana has now been shown to have strong antioxidative and neuroprotective effects, which may prolong neuronal cell survival. In areas where it is legal to do so, marijuana should be considered in the pharmacological management of ALS. Further investigation into the usefulness of marijuana in this setting is warranted.


Antithrombotic therapy for stroke prevention in atrial fibrillation and mechanical heart valves.
Cardioembolic strokes account for one-sixth of all strokes and are an important potentially preventable cause of morbidity and mortality. Vitamin K antagonists (e.g., warfarin) are effective for the prevention of cardioembolic stroke in patients with atrial fibrillation (AF) and in those with mechanical heart valves but because of their inherent limitations are underutilized and often suboptimally managed. Antiplatelet therapies have been the only alternatives to warfarin for stroke prevention in AF but although they are safer and more convenient they are much less efficacious. The advent of new oral anticoagulant drugs offers the potential to reduce the burden of cardioembolic stroke by providing access to effective, safe, and more convenient therapies. New oral anticoagulants have begun to replace warfarin for stroke prevention in some patients with AF, based on the favorable results of recently completed phase III randomized controlled trials, and provide for the first time an alternative to antiplatelet therapy for patients deemed unsuitable for warfarin. The promise of the new oral anticoagulants in patients with mechanical heart valves is currently being tested in a phase II trial. If efficacy and safety are demonstrated, the new oral anticoagulants will provide an alternative to warfarin for patients with mechanical heart valves and may also lead to increased use of mechanical valves for patients who would not have received them in the past because of the requirement for long term warfarin therapy.


[Effect of light of various spectral composition on kinetics of changes in a green leaf resistivity].
The kinetics of changes in the resistance of a green leaf by the action of exciting light was studied. It was shown that the illumination-induced changes in leaf conductance are mainly due to an elevation of temperature caused by the absorption of light energy. Based on the experimental results, the fraction of absorbed light energy converted into heat calculated under assumption that heating of the leaf across the thickness is uniform. It was found that the fraction of absorbed energy converted into heat does not depend on the intensity of radiation in the IR range and increases with decreasing intensity of photosynthetically active radiation. An explanation of the phenomenon is proposed, which is based on that in reality the heating of the leaf across the thickness is nonhomogeneous and this inhomogeneity of heating becomes more pronounced upon photoinduced changes in absorption resulting from a decrease in photosynthetically active radiation. It was shown that nonphotochemical fluorescence quenching does not substantially affect the leaf conductance at intensities of photosynthetically active radiation not higher than 200 Wt/m2.


The role of Golgi bodies in polysaccharide sulphation in Fucuszygotes.
The cell wall of 24-h zygotes of Fucus serratus is composed of 3 layers--an inner fibrillar layer (sulphated fucan), an outer fibrillar layer (alginic aicd/cellulose) and an exterior amorphous layer (sulphated fucan, alginic acid). The 2 layers containing sulphated fucan are preferentially thickened at the rhizoid pole. Light- and electron-microscope autoradiographic pulse-chase experiments on 22-h zygotes using 35SO2-(4) show the Golgi bodies to be the sites of fucan sulphation. The isolation and characterization of isolated Golgi-rich fractions from 22-h zygotes shows that the first detectable labelled macromolecule is associated with these fractions 2 min after addition of 35SO2-(4). The sulphate acceptor molecule has been partially characterized. 35S-APS and 35S-paps are detectable in the soluble fraction 0.5 min after addition of 35SO2-(4). The results are discussed in relation to other published work on the differentiation of Fucus embryos and on polysaccharide sulphation.


Gastric mechanosensory and lower esophageal sphincter function in rumination syndrome.
Our hypothesis was that rumination syndrome is associated with gastric sensory and motor dysfunction. We studied gastric and somatic sensitivity, reflex relaxation of the lower esophageal sphincter (LES), and gastric compliance and accommodation postprandially and postglucagon. A barostatically controlled gastric bag and esophageal manometry were used to compare gastric sensorimotor functions and LES relaxation to gastric distension in 12 patients with rumination syndrome and 12 controls. During bag distensions, patients had greater nausea, bloating, and aggregate score, but not pain, compared with controls (P < 0.05). At 4 and 8 mmHg gastric distension, LES tone reduction was greater in patients than in controls (P < 0.05). Gastric compliance, accommodation to a standard meal, and response to glucagon were not different in patients and controls; however, 6 of 12 patients had no gastric accommodation; the latter patients had significantly greater pain perception during distension (P < 0.05) but normal somatic sensitivity compared with healthy controls. Rumination syndrome is characterized by higher gastric sensitivity and LES relaxation during gastric distension. A subgroup of patients also had absent postprandial accommodation.


Multiplex chromatin interactions with single-molecule precision.
The genomes of multicellular organisms are extensively folded into 3D chromosome territories within the nucleus1. Advanced 3D genome-mapping methods that combine proximity ligation and high-throughput sequencing (such as chromosome conformation capture, Hi-C)2, and chromatin immunoprecipitation techniques (such as chromatin interaction analysis by paired-end tag sequencing, ChIA-PET)3, have revealed topologically associating domains4 with frequent chromatin contacts, and have identified chromatin loops mediated by specific protein factors for insulation and regulation of transcription5-7. However, these methods rely on pairwise proximity ligation and reflect population-level views, and thus cannot reveal the detailed nature of chromatin interactions. Although single-cell Hi-C8 potentially overcomes this issue, this method may be limited by the sparsity of data that is inherent to current single-cell assays. Recent advances in microfluidics have opened opportunities for droplet-based genomic analysis9 but this approach has not yet been adapted for chromatin interaction analysis. Here we describe a strategy for multiplex chromatin-interaction analysis via droplet-based and barcode-linked sequencing, which we name ChIA-Drop. We demonstrate the robustness of ChIA-Drop in capturing complex chromatin interactions with single-molecule precision, which has not been possible using methods based on population-level pairwise contacts. By applying ChIA-Drop to Drosophila cells, we show that chromatin topological structures predominantly consist of multiplex chromatin interactions with high heterogeneity; ChIA-Drop also reveals promoter-centred multivalent interactions, which provide topological insights into transcription.


Parallel firehose instability in electron-positron plasmas.
In a magnetized uniform plasma, firehose instability may arise as a result of pressure anisotropy of P_{||}>P_{⊥}, where P_{||} and P_{⊥} are the thermal pressure parallel and perpendicular to the magnetic field, respectively. In this paper, we examine the parallel firehose instability in electron-positron plasmas based on the particle simulations along with the linear fluid theory, which may give rise to the dispersion relation, instability criteria, and growth rate, etc., for comparisons with those calculated from the kinetic simulations. As for the firehose instability in electron-proton plasmas, the magnetic field grows rapidly and then decreases with oscillations. The nonlinear saturated state complies with the linear stability criterion, α=μ_{0}(P_{||}-P_{⊥})/B^{2}=1, derived from the fluid theory only for relatively smaller values of initial α or ω_{p}/ω_{c}, where ω_{p} and ω_{c} are the plasma and cyclotron frequencies, respectively. For relatively larger values of initial α and ω_{p}/ω_{c}, the saturated α values are smaller than 1 as a result of kinetic resonant effects. The dominant wave numbers are kc/ω_{p}<0.5 and the growth rates are in the range of 0.1-0.3ω_{c}, which are approximately consistent with the linear fluid theory for the same wavelengths. Both electrostatic and electromagnetic modes predicted by the linear fluid theory are identified in the kinetic simulations.


Signal sequence conservation and mature peptide divergence within subgroups of the murine beta-defensin gene family.
Beta-defensins are two exon genes which encode broad spectrum antimicrobial cationic peptides. We have analyzed the largest murine cluster of these genes which localizes to chromosome 8. Using hidden Markov models, we identified six beta-defensin exon 2-like sequences and subsequently found full-length expressed transcripts for these novel genes. Expression was high in brain and reproductive tissues. Eleven beta-defensins could be grouped into two clear subgroups by virtue of their position and high signal sequence (exon 1 encoded) identity. In contrast, however, there was a very low level of sequence conservation in the exon 2 region encoding the mature antimicrobial peptide. Examination of the gene sequences of orthologs in other rodents also revealed an excess of nucleotide changes that altered amino acids in the mature peptide region. Evolutionary analysis revealed strong evidence that following gene duplication, exon 1 and surrounding noncoding DNA show little divergence within subgroups. The focus for rapid sequence divergence is localized in the DNA encoding the mature peptide and this is driven by accelerated positive selection. This mechanism of evolution is consistent with the role of this gene family as defense against bacterial pathogens and the sequence changes have implications for novel antibiotic design.


Regulation of Akt/PKB activation by tyrosine phosphorylation.
Activation of Akt/PKB by growth factors requires multiple phosphorylation events. Phosphorylation of Thr(308) and Ser(473) of Akt by its upstream kinase(s) or autophosphorylation is critical for optimal activation of its kinase activity. Here, we present evidence that tyrosine phosphorylation is required for Akt activation. Epidermal growth factor treatment induces tyrosine phosphorylation of Akt in COS1 and PC3M cells, which is abrogated by PP2, a selective inhibitor for Src family tyrosine kinases. Elevated Akt activity is observed in v-Src transformed NIH3T3 cells, which is accompanied with increased tyrosine phosphorylation of Akt. Akt activity induced by growth factors is significantly reduced in SYF cells lacking Src, Yes, and Fyn, which can be restored by introducing c-Src, but not the kinase-inactive Src, back to these cells. Furthermore, we have identified two tyrosine residues near the activation loop of Akt that are important for its activation. Substitution of these residues with phenylalanine abolishes Akt kinase activity stimulated by growth factors. These two YF mutants fail to block Forkhead transcription factor activity in 293 cells and are unable to prevent apoptosis induced by matrix detachment. Our data suggest that, in addition to phosphorylation of Thr(308) and Ser(473), tyrosine phosphorylation of Akt may be essential for its biological function.


Negative differential resistance in carbon nanotube field-effect transistors with patterned gate oxide.
We demonstrate controllable and gate-tunable negative differential resistance in carbon nanotube field-effect transistors, at room temperature and at 4.2 K. This is achieved by effectively creating quantum dots along the carbon nanotube channel by patterning the underlying, high-kappa gate oxide. The negative differential resistance feature can be modulated by both the gate and the drain-source voltage, which leads to more than 20% change of the current peak-to-valley ratio. Our approach is fully scalable and opens up a possibility for a new class of nanoscale electronic devices using negative differential resistance in their operation.


Chronic urticaria from isophane insulin therapy; sensitivity associated with noninsulin components in commercial preparations.
A young woman with diabetes mellitus developed chronic urticaria after changing from isophane been insulin suspension to isophane beef-pork insulin suspension. She reverted to treatment with her original insulin preparation, but urticaria failed to terminate. While in the hospital, her eruption began each afternoon at the site of insulin injection. Zinc single-peak beef insulin suspension, a purer preparation with different additives than isophane beef insulin, was substituted, and urticaria terminated rapidly. Intradermal skin testing using single-peak (purified) preparations indicated that the patient was sensitive to beef and pork forms of isophane insulin but not to beef and pork forms of zinc insulin. The patient later had a brief recurrence of urticaria following oral erythromycin and tetracycline therapy but did not develop lesions at sites of insulin injection.


Cloning and analysis of the leuB gene of Leptospira interrogans serovar pomona.
The leuB gene of Leptospira interrogans serovar pomona strain kenniwicki has been cloned on a 9.5 kb plasmid, pWVL1, by complementation of Escherichia coli leuB mutants. Subcloning and Tn5 mutagenesis showed that the region required for complementation was approximately 1.2 kb in length. Enzyme assays showed that the product of the cloned gene was a beta-isopropylmalate dehydrogenase. Defects in the leuA, leuC and leuD genes of E. coli were not complemented by pWVL1. The nucleotide sequence of the leuB-complementing region and surrounding DNA has been determined. Three open reading frames were found which encode proteins of 40.9, 38.8 and 15 kDa. Analysis of subclones containing nucleotide deletions of varying sizes showed that only the 38.8 kDa protein was necessary to obtain complementation of E. coli leuB mutations. The PIR data base was searched and the enzyme 3-isopropylmalate dehydrogenase from six different micro-organisms was found to share significant amino acid sequence similarity (43-57%) with the 38.8 kDa L. interrogans leuB gene product. The organization of the leucine biosynthetic genes in L. interrogans differs from that found in E. coli, Salmonella typhimurium and Bacillus subtilis.


Construction and analysis of a Noccaea caerulescens TILLING population.
Metals such as Zn or Cd are toxic to plant and humans when they are exposed in high quantities through contaminated soil or food. Noccaea caerulescens, an extraordinary Zn/Cd/Ni hyperaccumulating species, is used as a model plant for metal hyperaccumulation and phytoremediation studies. Current reverse genetic techniques to generate mutants based on transgenesis is cumbersome due to the low transformation efficiency of this species. We aimed to establish a mutant library for functional genomics by a non-transgenic approach, to identify mutants with an altered mineral profiling, and to screen for mutations in bZIP19, a regulator of Zn homeostasis in N. caerulescens. To generate the N. caerulescens mutant library, 3000 and 5000 seeds from two sister plants of a single-seed recurrent inbred descendant of the southern French accession Saint-Félix-de-Pallières (SF) were mutagenized respectively by 0.3 or 0.4% ethyl methane sulfonate (EMS). Two subpopulations of 5000 and 7000 M2 plants were obtained after 0.3 or 0.4% EMS treatment. The 0.4% EMS treatment population had a higher mutant frequency and was used for TILLING. A High Resolution Melting curve analysis (HRM) mutation screening platform was optimized and successfully applied to detect mutations for NcbZIP19, encoding a transcription factor controlling Zn homeostasis. Of four identified point mutations in NcbZIP19, two caused non-synonymous substitutions, however, these two mutations did not alter the ionome profile compared to the wild type. Forward screening of the 0.4% EMS treatment population by mineral concentration analysis (ionomics) in leaf material of each M2 plant revealed putative mutants affected in the concentration of one or more of the 20 trace elements tested. Several of the low-Zn mutants identified in the ionomic screen did not give progeny, illustrating the importance of Zn for the species. The mutant frequency of the population was evaluated based on an average of 2.3 knockout mutants per tested monogenic locus. The 0.4% EMS treatment population is effectively mutagenized suitable for forward mutant screens and TILLING. Difficulties in seed production in low Zn mutants, obtained by both forward and reverse genetic approach, hampered further analysis of the nature of the low Zn phenotypes.


Neuropeptide-Y Y5 receptors modulate the onset and maintenance of operant ethanol self-administration.
Neuropeptide Y (NPY) is the most abundant and widely distributed peptide in the mammalian central nervous system and increases feeding behavior at NPY Y1 or Y5 receptor subtypes. Recent pharmacological and mutant mouse data indicate that NPY activity at its receptors can influence ethanol self-administration, although the direction and strength of this influence are not clear. Effects of the novel NPY Y5 receptor antagonist L-152,804 on the onset and maintenance of operant self-administration were examined in male C57BL/6J mice, which were trained to self-administer ethanol (10% v/v) versus water via the sucrose substitution method during 16 hr overnight sessions. After 4 months of baseline responding, mice were injected with L-152,804 (0, 10, 30, or 60 mg/kg, intraperitoneally) before operant sessions. Potential locomotor effects of L-152,804 and possible interaction with the sedative properties of ethanol also were examined. All three doses of L-152,804 significantly delayed the onset of ethanol-reinforced responding relative to vehicle injection. L-152,804 produced no effect on the total number of ethanol- or water-reinforced responses per 16 hr session. However, L-152,804 selectively modulated the temporal distribution of ethanol-reinforced responding depending on the dose (10 and 60 mg/kg) and time point measured in a manner consistent with blockade of ethanol reinforcement. Additional experiments determined that L-152,804 (10 or 60 mg/kg) did not alter spontaneous locomotor activity or influence the sedative effects of ethanol (4 g/kg). These results indicate that blockade NPY Y5 receptor activity modulates the onset and maintenance of ethanol self-administration. For this reason, NPY-Y5 receptor antagonists may be useful in medical management of alcohol abuse and alcoholism.


Fungal G-Protein-Coupled Receptors: A Promising Mediator of the Impact of Extracellular Signals on Biosynthesis of Ochratoxin A.
G-protein-coupled receptors (GPCRs) are transmembrane receptors involved in transducing signals from the external environment inside the cell, which enables fungi to coordinate cell transport, metabolism, and growth to promote their survival, reproduction, and virulence. There are 14 classes of GPCRs in fungi involved in sensing various ligands. In this paper, the synthesis of mycotoxins that are GPCR-mediated is discussed with respect to ligands, environmental stimuli, and intra-/interspecific communication. Despite their apparent importance in fungal biology, very little is known about the role of ochratoxin A (OTA) biosynthesis by Aspergillus ochraceus and the ligands that are involved. Fortunately, increasing evidence shows that the GPCR that involves the AF/ST (sterigmatocystin) pathway in fungi belongs to the same genus. Therefore, we speculate that GPCRs play an important role in a variety of environmental signals and downstream pathways in OTA biosynthesis. The verification of this inference will result in a more controllable GPCR target for control of fungal contamination in the future.


New treatment options for lipid-lowering therapy in subjects with type 2 diabetes.
Dyslipidemias represent a variety of quantitative and/or qualitative lipoprotein abnormalities. According to etiology, we distinguish primary dyslipidemias with strictly genetic background and secondary ones with their origin in other disease or pathological states. Diabetic dyslipidemia is a type of secondary dyslipidemia and plays an important role in determining the cardiovascular risk of subjects with type 2 diabetes. In these patients, insulin resistance is responsible for overproduction and secretion of atherogenic very low density lipoprotein. In addition, insulin resistance promotes the production of small dense low-density lipoprotein (LDL) and reduces high-density lipoprotein (HDL) production. Cardiovascular disease remains a leading cause of morbidity and mortality in diabetic patients. Previous results support the role for small, dense LDL particles in the etiology of atherosclerosis and their association with coronary artery disease. Moreover, lowering LDL cholesterol reduces the risk of cardiovascular death. Therefore, the European guidelines for the management of dyslipidemias recommend an LDL cholesterol goal < 100 mg/dL in diabetic subjects without cardiovascular events. Moreover, if triglycerides (TG) are elevated (> 400 mg/dL), they recommend a non-HDL cholesterol goal < 130 mg/dL in diabetic individuals without cardiovascular events. Statins are the first line of LDL-lowering therapy in diabetic patients and combined therapy with ezetimibe and statins could be useful in very high cardiovascular risk diabetic subjects. Furthermore, the effect of a fibrate as an add-on treatment to a statin could improve the lipid profile in diabetic individuals with high TG and low HDL cholesterol. Regarding new therapies, recent data from phase III trials show that proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors considerably decrease LDL cholesterol. Thus, they may be useful in diabetic patients with concomitant diseases such as familial dyslipidemia, recurrent cardiovascular events, and elevated LDL cholesterol after second drug administration in addition to maximal statin dose or statin intolerance.


The putative drug efflux systems of the Bacillus cereus group.
The Bacillus cereus group of bacteria includes seven closely related species, three of which, B. anthracis, B. cereus and B. thuringiensis, are pathogens of humans, animals and/or insects. Preliminary investigations into the transport capabilities of different bacterial lineages suggested that genes encoding putative efflux systems were unusually abundant in the B. cereus group compared to other bacteria. To explore the drug efflux potential of the B. cereus group all putative efflux systems were identified in the genomes of prototypical strains of B. cereus, B. anthracis and B. thuringiensis using our Transporter Automated Annotation Pipeline. More than 90 putative drug efflux systems were found within each of these strains, accounting for up to 2.7% of their protein coding potential. Comparative analyses demonstrated that the efflux systems are highly conserved between these species; 70-80% of the putative efflux pumps were shared between all three strains studied. Furthermore, 82% of the putative efflux system proteins encoded by the prototypical B. cereus strain ATCC 14579 (type strain) were found to be conserved in at least 80% of 169 B. cereus group strains that have high quality genome sequences available. However, only a handful of these efflux pumps have been functionally characterized. Deletion of individual efflux pump genes from B. cereus typically had little impact to drug resistance phenotypes or the general fitness of the strains, possibly because of the large numbers of alternative efflux systems that may have overlapping substrate specificities. Therefore, to gain insight into the possible transport functions of efflux systems in B. cereus, we undertook large-scale qRT-PCR analyses of efflux pump gene expression following drug shocks and other stress treatments. Clustering of gene expression changes identified several groups of similarly regulated systems that may have overlapping drug resistance functions. In this article we review current knowledge of the small molecule efflux pumps encoded by the B. cereus group and suggest the likely functions of numerous uncharacterised pumps.


[Combination of anxiety disorder and alcohol dependence].
Three patients, two men aged 47 and 37 and a woman aged 32, are described. All three suffered from a combination of alcohol addiction and anxiety disorder. None of them had previously received adequate treatment because on the one hand they had been refused by psychiatric institutes because of their alcohol addiction, and on the other the institutes for the care of addicted had not diagnosed the anxiety disorder. About 150,000 people are estimated to have both an alcohol addiction and an anxiety disorder in the Netherlands. The importance of making a correct diagnosis and providing proper treatment for both disorders is emphasized.


Matrix-Bound Zolzoledronate Enhances the Biofilm Colonization of Hydroxyapatite: Effects on Osteonecrosis.
(1) Background: The aim of this study was to test whether matrix-bound zoledronate (zol) molecules enhanced the oral biofilm colonization of a mineralized matrix, rendering the alveolar bone more susceptible to medication-related osteonecrosis of the jaw (MRONJ) following invasive dental procedures. (2) Methods: We tested the effect of matrix-bound zol on the growth and attachment of Porphyromonas gingivalis (Pg), Fusobacterium nucleatum (Fn) and Actinomyces israelii (Ai), and whether the nitrogen-containing component of zol contributed to such effect. The role of oral bacteria in the induction of osteonecrosis was then tested using an extra-oral bone defect model. (3) Results: The attachment of biofilm to hydroxyapatite discs increased when the discs were pre-treated with zol. Bacterial proliferation was not affected. Matrix-bound zol was more potent than non-nitrogen-containing etidronate in enhancing the colonization. Stimulation was dampened by pre-treating the bacteria with histidine. The delivery of oral biofilm to a tibial defect caused osteonecrosis in zol-treated rats. (4) Conclusions: We conclude that matrix-bound zol enhances the oral biofilm colonization of hydroxyapatite. This enhancement depended on the presence of the nitrogen-containing group. The oral biofilm rendered the extra-oral bone susceptible to medication-related osteonecrosis, suggesting that it has an important role in the induction of MRONJ.


Factors influencing mothers' role in convulsion treatment among under-five children in Ibadan, Nigeria.
Convulsion among children between six months and five years is a major contributor to childhood mortality in less-developed societies, especially in sub-Saharan Africa. Most studies on under-five deaths have ignored the influence of socio-demographic and environmental factors as they relate to causes of the thematic health condition and available therapies. This study investigated mothers' perception of convulsion causation, relevant signs and symptoms, and the influence of socio-economic status on mothers' choice of remedies. The research was conducted in Ibadan, southwest Nigeria, which is densely populated with mainly Yoruba-speaking people. The study population comprised mothers who, at the time of fieldwork for the present analysis, had at least one under-five child. Five hundred questionnaire respondents were selected through a multistage sampling technique, and 14 in-depth interviews (IDIs) were conducted among different categories of women identified through the snowball technique. Voluntary Social Action Theory and the Health Belief Model were used in explaining the relationship between the dependent and independent variables. The findings show that the remedies mothers chose were strongly influenced by socio-demographic factors such as marriage type, religion, level of education, occupation and place of residence. It is strongly suggested that interventions, health policies and programs should focus on how best to empower women to effectively utilize medical information that will enable them recognize symptoms of this common health condition and/or undertake preliminary therapies that contribute positively to convulsion prevention or treatment.


Estrogen protection against atherosclerosis and synthetic estrogen production of cirrhosis in the rabbit.
Two artificial estrogens, ethinyl estradiol and mestranol, were found to cause cirrhosis in the rabbit liver during a study of atherosclerosis. These drugs showed protective effects against atherosclerosis in cholesterol-fed rabbits when administered orally (1 ppm in diet). These drugs and two naturally occurring estrogens, estrone and estradiol, were similarly effective when administered intramuscularly (1.5 mg per rabbit per week) to rabbits fed a diet lower in cholesterol. Reduction by estrogens of plasma cholesterol did not fully account for the reduction of extent of aortic atherosclerosis. Also, no differences in aortic or platelet eicosanoid production from exogenous arachidonic acid were found to explain the effect on atherogenesis. The 2 artificially modified estrogens, ethinyl estradiol and mestranol, caused portal fibrosis with biliary proliferation (even in rabbits not fed cholesterol). Estrone, estradiol, and testosterone were not injurious in this regard at the dosages used. Only the unmodified estrogens reduced atherosclerosis without damaging the rabbit liver.


Does L-carnitine supplementation affect serum levels of enzymes mainly produced by liver? A systematic review and meta-analysis of randomized controlled clinical trials.
L-carnitine supplementation is proposed to reduce liver enzymes levels; however, previous findings were equivocal. The current systematic review and meta-analysis of randomized controlled clinical trials (RCTs) were performed to assess the effect of L-carnitine supplementation on serum levels of enzymes mainly produced by liver [alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyl transpeptidase (GGTP)]. Online databases as well as the reference lists of relevant studies were searched from inception up to June 2019. The risk of bias in individual studies was assessed using Cochrane Collaboration's tool. Data were pooled using the random-effects model and expressed as mean differences (MDs) with 95% confidence intervals (CIs). In total, 18 RCTs (1161 participants) met the eligibility criteria. L-carnitine supplementation dose ranged from 500 to 4000 mg/day. L-carnitine supplementation significantly reduced serum ALT (MD = - 8.65 IU/L, 95% CI - 13.40, - 3.90), AST (MD = - 8.52 IU/L, 95% CI - 12.16, - 4.89), and GGTP (MD = - 8.80 IU/L, 95% CI - 13.67, - 3.92) levels. The subgroup analysis showed that L-carnitine might be more effective in reducing the enzymes when supplemented in higher doses (≥ 2000 mg/day), for longer durations (> 12 weeks), and among patients with liver diseases. The meta-evidence was graded as "moderate" for ALT and AST, and "low" for GGTP according to NutriGrade scoring system. L-carnitine supplementation significantly improves circulating ALT, AST and GGTP levels; therefore, it might positively affect liver function, especially among patients with liver diseases. Further high-quality RCTs are recommended to confirm our results.


Insulin-like growth factor binding protein 2: an androgen-dependent predictor of prostate cancer survival.
Evidence suggests that the insulin-like growth factor (IGF) family is important in prostate cancer. We evaluate the ability of IGF markers to predict biochemical recurrence-free survival (bRFS) following radical prostatectomy (RRP). Preoperative sera from 141 patients undergoing RRP were analyzed for IGF-I, IGFBP-1, IGFBP-2 and IGFBP-3. A multivariate Cox model was created to assess the ability of these markers to predict bRFS. Preoperative covariables included: biopsy Gleason score, clinical TNM stage, serum PSA and neoadjuvant hormonotherapy. Kaplan-Meier curves were stratifying by IGF cutpoints (determined by ROC analysis) and hormonotherapy status. Average follow-up was 6.92 years, median PSA was 6.9 ng/ml and 85.1% of patients had cT2NxM0 disease. 49 patients experienced a PSA failure. Average levels of IGF-I, IGFBP-1, IGFBP-2 and IGFBP-3 were 156.5 ng/ml, 48.1 ng/ml, 396.8 ng/ml and 3303.7 ng/ml, respectively. IGFBP-2 was an independent predictor of PSA failure. Patients treated with neoadjuvant ADT (n=71) and whom had high IGFBP-2 levels experienced a 5-year bRFS that was better than those with low IGFBP-2 levels (77.7% vs. 53.3%). Patients without neoadjuvant ADT whom had high IGFBP-2 levels had a 5-year bRFS that was worse than those with low IGFBP-2 levels (64.5% vs. 82.7%). Preoperative IGFBP-2 predicts post-radical prostatectomy bRFS and is independent of stage, Gleason score and PSA. Increased IGFBP-2 is associated with better survival in patients with neoadjuvant hormonotherapy but worse survival in those without, suggesting a dramatic switch in function of this protein dependent on the presence or absence of androgens.


Effect of recombinant human nerve growth factor eye drops in patients with dry eye: a phase IIa, open label, multiple-dose study.
Dry eye disease (DED) affects more than 14% of the elderly population causing decrease of quality of life, high costs and vision impairment. Current treatments for DED aim at lubricating and controlling inflammation of the ocular surface. Development of novel therapies targeting different pathogenic mechanisms is sought-after. The aim of this study is to evaluate safety and efficacy of recombinant human nerve growth factor (rhNGF) eye drops in patients with DED. Forty consecutive patients with moderate to severe DED were included in a phase IIa, prospective, open label, multiple-dose, clinical trial to receive rhNGF eye drops at 20 µg/mL (Group 1: G1) or at 4 µg/mL (Group 2: G2) concentrations, two times a day in both eyes for 28 days (NCT02101281). The primary outcomes measures were treatment-emerged adverse events (AE), Symptoms Assessment in Dry Eye (SANDE) scale, ocular surface staining and Schirmer test. Of 40 included patients, 39 completed the trial. Both tested rhNGF eye drop concentrations were safe and well tolerated. Twenty-nine patients experienced at least one AE (14 in G1 and 15 in G2), of which 11 had at least 1 related AE (8 in G1 and 3 in G2). Both frequency and severity of DED symptoms and ocular surface damage showed significant improvement in both groups, while tear function improved only in G1. The data of this study indicate that rhNGF eye drops in both doses is safe and effective in improving symptoms and signs of DED. Randomised clinical trials are ongoing to confirm the therapeutic benefit of rhNGF in DED. NCT02101281.


Sequence and structural analysis of the chitinase insertion domain reveals two conserved motifs involved in chitin-binding.
Chitinases are prevalent in life and are found in species including archaea, bacteria, fungi, plants, and animals. They break down chitin, which is the second most abundant carbohydrate in nature after cellulose. Hence, they are important for maintaining a balance between carbon and nitrogen trapped as insoluble chitin in biomass. Chitinases are classified into two families, 18 and 19 glycoside hydrolases. In addition to a catalytic domain, which is a triosephosphate isomerase barrel, many family 18 chitinases contain another module, i.e., chitinase insertion domain. While numerous studies focus on the biological role of the catalytic domain in chitinase activity, the function of the chitinase insertion domain is not completely understood. Bioinformatics offers an important avenue in which to facilitate understanding the role of residues within the chitinase insertion domain in chitinase function. Twenty-seven chitinase insertion domain sequences, which include four experimentally determined structures and span five kingdoms, were aligned and analyzed using a modified sequence entropy parameter. Thirty-two positions with conserved residues were identified. The role of these conserved residues was explored by conducting a structural analysis of a number of holo-enzymes. Hydrogen bonding and van der Waals calculations revealed a distinct subset of four conserved residues constituting two sequence motifs that interact with oligosaccharides. The other conserved residues may be key to the structure, folding, and stability of this domain. Sequence and structural studies of the chitinase insertion domains conducted within the framework of evolution identified four conserved residues which clearly interact with the substrates. Furthermore, evolutionary studies propose a link between the appearance of the chitinase insertion domain and the function of family 18 chitinases in the subfamily A.


An LCA researcher's wish list--data and emission models needed to improve LCA studies of animal production.
The last decade has seen an increase in environmental systems analysis of livestock production, resulting in a significant number of studies with a holistic approach often based on life-cycle assessment (LCA) methodology. The growing public interest in global warming has added to this development; guidelines for carbon footprint (CF) accounting have been developed, including for greenhouse gas (GHG) accounting of animal products. Here we give an overview of methods for estimating GHG emissions, with emphasis on nitrous oxide, methane and carbon from land use change, presently used in LCA/CF studies of animal products. We discuss where methods and data availability for GHGs and nitrogen (N) compounds most urgently need to be improved in order to produce more accurate environmental assessments of livestock production. We conclude that the top priority is to improve models for N fluxes and emissions from soils and to implement soil carbon change models in LCA/CF studies of animal products. We also point at the need for more farm data and studies measuring emissions from soils, manure and livestock in developing countries.


A monoclonal antibody for distinction of invasive and noninvasive clinical isolates of Entamoeba histolytica.
Approximately 10% of the world population is infected with Entamoeba histolytica, but only 10% of the carriers develop symptomatic amebiasis. This discrepancy could be explained by the genotypic differences between the morphologically indistinguishable invasive and noninvasive strains of E. histolytica currently identified by zymodeme analysis, a technique that is unsuitable for routine diagnostic laboratories. Here we report the production of a monoclonal antibody against E. histolytica and its use in an immunofluorescence assay to identify invasive isolates cultured from stool samples of infected patients in several regions where amebiasis is endemic: Bangladesh, Colombia, and Mexico. After testing a total of 88 E. histolytica isolates, the correlation between zymodeme characterization and the immunofluorescence assay with the invasive isolate-specific monoclonal antibody was 100%. The epitope detected by the invasive isolate-specific monoclonal antibody resides in a previously undescribed internal protein with molecular masses of 84 and 81 kDa in axenic and polyxenic E. histolytica strains, respectively.


[Open wedge osteotomy of the tibial head. Management of vascular complications].
Lesions of the popliteal artery during high tibial osteotomy are rare complications, consequently the majority of publications are case related. The interval between surgery and diagnosis is reported to be as long 3 years; therefore, the current literature probably does not reflect the true incidence of vascular injuries. The case reports published in the literature were further evaluated. The focus was on the normal vascular anatomy of the popliteal region and anatomical deviations that predispose to vascular injury. As the flexion angle of the knee joint is considered to be decisive for vascular injury, this aspect was also an additional focus. For the unlikely event of a vascular injury, recommendations are presented which indicate diagnostic and therapeutic decisions. We analyzed the available literature and present own magnetic resonance imaging (MRI) investigations of the popliteal artery with different angles of flexion in six healthy volunteers. A variation of the origin of the anterior tibial artery with a course between the posterior tibial cortex and the popliteal muscle was found in 6% of all patients and predisposes to an accidental injury during osteotomy. The results in the literature and our own MRI findings suggest that a flexion angle of 90° facilitates anatomical dissection and osteotomy but cannot be regarded as a reliable protection against vascular injury.


Clinical characteristics of unipolar disorder and bipolar disorder according to the lifetime presence of recurrent panic attacks.
The frequent comorbidity of panic and affective disorders has been described in previous studies. However, it is not clear how panic disorder comorbidity in unipolar disorder and bipolar disorder is related to illness course. We compared lifetime clinical characteristics of illness and items of symptomatology in samples of individuals with bipolar I disorder (n = 290) and unipolar disorder (n = 335) according to the lifetime presence of recurrent panic attacks. We found significant differences in clinical course of illness characteristics that were shared across the unipolar and bipolar samples according to the lifetime presence of panic attacks. We also found a number of differences according to the presence of panic attacks that may be specific to the diagnostic group. Distinguishing patients who have mood disorder diagnoses, especially bipolar I disorder, according to the lifetime presence of panic attacks may not only be of use in clinical practice, but may also be informative for aetiological research, such as molecular genetic studies.


Pathogenicity determinants and antibiotic resistance profiles of enterococci from foods of animal origin in Turkey.
In this study, the presence of genes responsible for the pathogenicity and antibiotic resistance profile of enterococci isolated from various foodstuffs of animal origin was investigated. The percentage prevalence of enterococci was 54.1% (203/375) and the average count was found to be 3.81 log cfu/ml-g. Species-specific primers revealed Enterococcus faecalis as the predominant species carrying one or more virulence-associated traits of efa, gelE, ace, esp and agg genetic markers. Only one E. faecium isolate (from milk) was positive for the esp gene. Regarding antibiotic resistance, the highest frequency of resistance was observed for tetracycline (21.7%), followed by quinupristin/dalfopristin (13.3%), ciprofloxacin (2.0%), penicillin (2.0%), linezolid (1.0%), ampicillin (1.0%), streptomycin (1.0%), and gentamicin (0.5%). Enterococcus faecalis showed a higher prevalence of antibiotic resistance than other enterococci. The percentage of multidrug resistance among the isolates was 3.4%. Twenty-nine E. faecalis isolates (26.6%) carrying one of the virulence-associated traits were at the same time resistant to at least one antibiotic. Our results show that foods of animal origin, including ready-to-eat products, may be reservoirs of antibiotic-resistant and potentially virulent enterococci.


Obesity and low back pain: a retrospective cohort study of Japanese males.
[Purpose] This study evaluated whether obesity is a risk factor for low back pain, by using body fat percentage (%FAT) and body mass index (BMI) as indices of obesity among Japanese males. [Subjects and Methods] This study included 1,152 males (average age: 28.0 ± 4.6 years). BMI was calculated from subject's height and weight, and %FAT was estimated by the thickness of two parts of skin. Low back pain, drinking and smoking were surveyed using a self-administered questionnaire, and maximal oxygen uptake was measured by a submaximal exercise test using a cycle ergometer. [Results] A significant positive dose-response relationship was shown between %FAT and persistent low back pain prevalence. Similarly, a significant positive dose-response relationship was confirmed between BMI and persistent low back pain. [Conclusion] This study suggests that both high %FAT and BMI are risk factors for persistent low back pain.


Design, optimization, and in vivo evaluation of invasome-mediated candesartan for the control of diabetes-associated atherosclerosis.
Atherosclerosis is an inflammatory disease characterized by the accumulation of arterial plaque. Diabetes mellitus stands out as a major risk factor for atherosclerosis. Candesartan is a potent angiotensin II receptor antagonist that enhances arterial blood flow and reduces insulin resistance. However, oral candesartan has limited activity because of its low bioavailability, water solubility, hepatic first-pass degradation, and efficacy. The current study aims to develop nasal candesartan-loaded invasome (CLI) drops to improve candesartan's permeation, release, and bioavailability as a potential treatment for diabetes-associated atherosclerosis. Design expert software was used to prepare various CLI formulations to determine the impact of the concentrations of ethanol, cineole, and phospholipid. The desirability index was used to estimate the optimized formulation composition to maximize entrapment efficiency and minimize vesicle size. The optimized formulation had a 1% ethanol concentration, a 1.5% cineole concentration, and a 2.32% phospholipid concentration. The selected optimized formulation was then tested in a rat model of diabetes and atherosclerosis to evaluate its activity. The results showed that nasal CLI drops significantly raised serum HDL levels by a ratio of 1.42 and lowered serum glucose, cholesterol, triglycerides, LDL, and VLDL levels by 69.70%, 72.22%, 36.52%, 58.0%, and 65.31%, respectively, compared with diabetic atherosclerotic rats, throwing an insight on the potential for promising anti-diabetic and anti-atherosclerotic activities. Additionally, atherosclerotic lesions were improved in rats treated with CLI, as shown in histopathology. In conclusion, the results of this investigation showed that treatment with nasal CSN-loaded invasome formulation drops prevented the initiation and progression of diabetes-associated atherosclerosis.


Recent development of organic-inorganic hybrid photocatalysts for biomass conversion into hydrogen production.
Over the last few years, photocatalysis using solar radiation has been explored extensively to investigate the possibilities of producing fuels. The production and systematic usage of solar fuels can reduce the use of fossil-based fuels, which are currently the primary source for the energy. It is time for us to exploit renewable sources for our energy needs to progress towards a low-carbon society. This can be achieved by utilizing green hydrogen as the future energy source. Solar light-assisted hydrogen evolution through photocatalytic water splitting is one of the most advanced approaches, but it is a non-spontaneous chemical process and restricted by a kinetically demanding oxidation evolution reaction. Sunlight is one of the essential sources for the photoreforming (PR) of biomass waste into solar fuels, or/and lucrative fine chemicals. Hydrogen production through photoreforming of biomass can be considered energy neutral as it requires only low energy to overcome the activation barrier and an alternate method for the water splitting reaction. Towards the perspective of sustainability and zero emission norms, hydrogen production from biomass-derived feedstocks is an affordable and efficient process. Widely used photocatalyst materials, such as metal oxides, sulphides and polymeric semiconductors, still possess challenges in terms of their performance and stability. Recently, a new class of materials has emerged as organic-inorganic hybrid (OIH) photocatalysts, which have the benefits of both components, with peculiar properties and outstanding energy conversion capability. This work examines the most recent progress in the photoreforming of biomass and its derivatives using OIHs as excellent catalysts for hydrogen evolution. The fundamental aspects of the PR mechanism and different methods of hydrogen production from biomass are discussed. Additionally, an interaction between both composite materials at the atomic level has been discussed in detail in the recent literature. Finally, the opportunities and future perspective for the synthesis and development of OIH catalysts are discussed briefly with regards to biomass photo-reforming.


Guidance signalling regulates leading edge behaviour during collective cell migration of cardiac cells in Drosophila.
Collective cell migration is the coordinated movement of cells, which organize tissues during morphogenesis, repair and some cancers. The motile cell membrane of the advancing front in collective cell migration is termed the Leading Edge. The embryonic development of the vertebrate and Drosophila hearts are both characterized by the coordinated medial migration of a bilateral cluster of mesodermal cells. In Drosophila, the cardioblasts form cohesive bilateral rows that migrate collectively as a unit towards the dorsal midline to form the dorsal vessel. We have characterized the collective cell migration of cardioblasts as an in vivo quantitative model to study the behaviour of the Leading Edge. We investigated whether guidance signalling through Slit and Netrin pathways plays a role in cell migration during heart development. Through time-lapse imaging and quantitative assessment of migratory behaviour of the cardioblasts in loss-of-function mutants, we demonstrate that both Slit and Netrin mediated signals are autonomously and concomitantly required to maximize migration velocity, filopodial and lamellipodial activities. Additionally, we show that another Slit and Netrin receptor, Dscam1, the role of which during heart development was previously unknown, is required for both normal migration of cardioblasts and luminal expansion. Leading edge behaviour analysis revealed a dosage dependent genetic interaction between Slit and Netrin receptors suggesting that downstream signalling through these receptors converge on a common output that increases leading edge activity of the cardioblasts. Finally, we found that guidance signalling maintains the balance between epithelial and mesenchymal characteristics of the migrating cardioblasts.


Effect of Mycobacterium leprae in peripheral nerve trunk on the evolution of skin lesions.
Comparative histological studies were made of a) 41 peripheral nerve lesions and the skin in the area of supply, and b) 12 peripheral nerve lesions and concurrent but unrelated skin lesions. In the first study, small, relatively early, histologically classifiable skin lesions were found in all cases, even though there were no clinical lesions. In every case the lesion was centered on a dermal nerve. In some cases disruption of the perineurium was associated with emergence of the lesion into the dermis and a small silent local reaction. It was concluded that there was a descending spread of the disease down the neural pathway to the dermis, although it was not necessarily associated with transport of bacilli. Although the first study showed a discrepancy in the classification between skin and nerve lesions in nearly 50% of the cases (as previously reported), the second study showed no discrepancies. It is suggested that discrepancies are relatively uncommon, and that those in the first study are exceptional. The probable explanation is that microreactions in the nerve trunks had caused a shift in classification, which was not yet reflected in the immature skin lesions. In the second study, the mature skin lesions had reached immunological equilibrium. Discrepancies in classification between skin and nerve lesions, as between concurrent skin lesions, are the result of reaction. Attention is drawn to the probable role of subliminal reactions in the evolution of infections.


Patching for corneal abrasion.
Recent audits show that corneal abrasion is a common presenting eye complaint. Eye patches are often recommended for treating corneal abrasions despite the lack of evidence for their use. This systematic review was conducted to determine the effects of the eye patch when used to treat corneal abrasions. The objective of this review was to test the hypothesis that patching an eye following a corneal abrasion improves healing or provides pain relief. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (which contains the Cochrane Eyes and Vision Group Trials Register) on The Cochrane Library (2005, Issue 2), MEDLINE (1966 to April 2005), EMBASE (1980 to April 2005), LILACS (13 April 2005), NRR (2005, Issue 2) and SIGLE (December 2004). There were no language or date restrictions in the searches. We also searched the reference lists of included studies, unpublished 'grey' literature and conference proceedings and contacted pharmaceutical companies for details of unpublished trials. We included randomised and quasi-randomised controlled trials that compared patching the eye with no patching to treat simple corneal abrasions. Two authors independently assessed trial quality and extracted data. We contacted investigators for further information regarding quality of trials. The primary outcome was healing of the corneal epithelium and secondary outcomes were related to pain. Eleven trials, which randomised a total of 1014 participants, were included in the review. Meta-analysis of seven studies with dichotomous healing outcomes favoured no patching on the first day of healing (risk ratio (RR) 0.89, 95% Confidence Interval (CI) 0.79 to 0.99). For days two and three there was no significant difference between the two groups. Of the nine trials that measured pain scores two favoured no patching and none favoured patching. Complication rates were low and no differences were noted in these between the two groups. No-patch groups generally received more adjuvant treatment with antibiotics and/or cycloplegics than the patch group which is an important confounding factor. Treating simple corneal abrasions with a patch does not improve healing rates on the first day post-injury and does not reduce pain. In addition, use of patches results in a loss of binocular vision. Therefore it is recommended that patches should not be used for simple corneal abrasions. Further research should focus on large (greater than 10 mm(2)) abrasions.


Poliovirus receptor (CD155) regulates a step in transendothelial migration between PECAM and CD99.
The movement of leukocytes across endothelium [referred to as diapedesis or transendothelial migration (TEM)] is a critical step in the inflammatory process. Recently, it was demonstrated that treatment of endothelial cells and monocytes with antibodies against poliovirus receptor (PVR; CD155) and DNAX-associated molecule-1 (DNAM-1; CD226) arrested monocytes over endothelial junctions and prevented TEM, suggesting that these molecules are involved in diapedesis. However, nothing was known about the mechanism by which PVR and DNAM-1 work in TEM. Herein, we show that, similar to endothelial PECAM interacting with leukocyte PECAM, activation of endothelial PVR with anti-PVR antibodies or interaction with its ligand, DNAM-1, results in recruitment of the tyrosine phosphatase Shp-2, and this process is dependent on Src kinases. Furthermore, differential and sequential treatment with blocking antibodies directed against PVR, DNAM-1, PECAM, and CD99 showed that endothelial PVR and monocyte DNAM-1 interact at and regulate a step between those regulated by PECAM and CD99. Further studies demonstrate that PVR resides in the recently identified lateral border recycling compartment, similar to PECAM and CD99. These findings suggest that the localization of adhesion/signaling molecules to the lateral border recycling compartment and the recruitment of Shp-2 may be common mechanisms for the regulation of TEM by endothelial cells.


Interference with activities due to pain and fatigue: accuracy of ratings across different reporting periods.
This study examined the impact of different reporting period lengths on the accuracy of items measuring interference due to pain and fatigue with work, walking, and relations with others. Six items from well-established instruments (Brief Pain Inventory, Brief Fatigue Inventory, SF-36) were investigated in a prospective study of 117 patients with chronic rheumatological illness. Daily ratings were compared with recall ratings of 1, 3, 7, and 28-day reporting periods. The level of recall ratings (RRs) for reporting periods of 3 days or more were significantly higher than the level of aggregated end-of-day (EOD) ratings. Correspondence between aggregated EOD and RRs was good (r ≥ .80) regardless of the length of the reporting period. Ratings of interference for a single day were highly correlated with aggregated EOD for up to 14 days prior to the single rating (r ≥ .76). Recall ratings with reporting periods of up to a month yield good correspondence with aggregated daily ratings, although the absolute level of the rating will be inflated for recall periods of 3 days or longer.


Magnetic resonance imaging in pulmonary hypertension: an overview of current applications and future perspectives.
Pulmonary hypertension is an heterogeneous group of diseases characterised by increased pulmonary arterial pressures which impact on the upstream right ventricle. Pulmonary hypertension can be challenging to diagnose, classify and monitor when specific therapies are applicable. Cardiac magnetic resonance (CMR) imaging has greatly evolved in the last decades and is a promising tool to non-invasively follow pulmonary hypertension patients. CMR provides a comprehensive evaluation of the heart and is therefore the gold standard for quantification of right ventricular volumes, mass and function, which are critical for pulmonary hypertension prognosis. In addition, innovative MR techniques allow an increasingly precise evaluation of pulmonary haemodynamics and lung perfusion. This review highlights the main advantages offered by CMR in pulmonary hypertension and gives an overview of putative future applications. Although right heart catheterisation remains mandatory in the diagnostic algorithm, CMR could play an increasingly important role in the coming years in monitoring pulmonary hypertension patients.


Endocrine pathogenesis of postweaning anestrus in swine: response of the persistently anestrous sow to hormonal stimuli.
The endocrine function of the individual components of the hypothalamo-hypophyseal-ovarian axis of the postweaning anestrous sow was evaluated by monitoring the sow's response to exogenous estradiol, gonadotropin releasing hormone (GnRH), and gonadotropins. Sows (4 to 6/group) not returning to estrus by 42.8 +/- 3.1 days after weaning were assigned to 1 of the following treatments: 10 micrograms of estradiol benzoate (EB)/kg of body weight; 200 micrograms of GnRH, 1,000 IU of pregnant mare's serum gonadotropin (PMSG); 1,000 IU of human chorionic gonadotropin (HCG); or 4 ml of saline solution plus 2 ml of corn oil. A preovulatory-like surge of luteinizing hormone [(LH) greater than 12 hours in duration] was observed in all weaned sows treated with EB. All EB-treated sows exhibited estrus and ovulated but none conceived. Sows given GnRH had transiently increased (less than 3 hours) LH concentrations that were not associated with estrus or ovulation. Treatment with PMSG caused an increase in peripheral concentrations of 17 beta-estradiol that was followed by an LH surge, estrus, ovulation, and conception. Treatment with HCG caused an increase in circulating concentrations of 17 beta-estradiol that was accompanied by a surge of LH in some sows and ovulation in all sows. Not all sows treated with HCG exhibited estrous behavior, but conception occurred in 2 of 3 sows that were mated at estrus. None of the sows treated with saline plus corn oil had consistent changes in circulatory concentrations of 17 beta-estradiol or LH and none exhibited estrus or ovulated.(ABSTRACT TRUNCATED AT 250 WORDS)


Interaction of PRP4 with Kruppel-like factor 13 regulates CCL5 transcription.
Activation of resting T lymphocytes initiates differentiation into mature effector cells over 3-7 days. The chemokine CCL5 (RANTES) and its major transcriptional regulator, Krüppel-like factor 13 (KLF13), are expressed late (3-5 days) after activation in T lymphocytes. Using yeast two-hybrid screening of a human thymus cDNA library, PRP4, a serine/threonine protein kinase, was identified as a KLF13-binding protein. Specific interaction of KLF13 and PRP4 was confirmed by reciprocal coimmunoprecipitation. PRP4 is expressed in PHA-stimulated human T lymphocytes from days 1 and 7 with a peak at day 3. Using an in vitro kinase assay, it was found that PRP4 phosphorylates KLF13. Furthermore, although phosphorylation of KLF13 by PRP4 results in lower binding affinity to the A/B site of the CCL5 promoter, coexpression of PRP4 and KLF13 increases nuclear localization of KLF13 and CCL5 transcription. Finally, knock-down of PRP4 by small interfering RNA markedly decreases CCL5 expression in T lymphocytes. Thus, PRP4-mediated phosphorylation of KLF13 plays a role in the regulation of CCL5 expression in T lymphocytes.


Oral Porphyromonas gingivalis infection affects intestinal microbiota and promotes atherosclerosis.
The link between periodontitis and intestinal dysbiosis, two factors that contribute to atherosclerosis, has not been clearly defined. We investigated the integrative effects of oral infection with Porphyromonas gingivalis (PG), the major pathogen for periodontitis, on intestinal microbiota and atherosclerosis. ApoE-/- mice were fed a normal chow diet (NC), a Western diet (WD) or a WD with oral PG infection (PG). The PG infection was investigated by placing a total of 109 CFUs of live PG into the oral cavity of each mouse using a feeding needle five times a week for 3 weeks. Atherosclerotic lesions of the aortae were measured, and blood lipoproteins and the expression of molecules related to lipid metabolism in the liver were analysed. We also performed 16S RNA sequencing and a microbiome analysis using faeces. En face bloc preparation of the aortae showed that the PG group had a 1.7-fold increase in atherosclerotic lesions compared with the WD group (p < .01). Serum analyses showed that oral PG infection induced a significant decrease in high-density lipoprotein (HDL) and triglyceride. Western blots of hepatic tissue lysates revealed that PG infection reduced the expression of scavenger receptor class B type 1 (SR-B1) in the liver by 50%. Faecal microbiota analysis revealed that species richness estimates (Chao1, ACE) decreased immediately after PG infection. PG infection also induced a significant decrease in Shannon diversity and an increase in Simpson's indices in the WD-fed mice. PG infection significantly increased the phyla Actinobacteria and Deferribacteres, along with the species Mucispirillum schaedleri and Lactobacillus gasseri, in the mice. The functional study showed that PG infection increased the expression of proteins that function in carbohydrate and glucose metabolism, including phosphotransferase system (PTS) proteins and the GntR family transcriptional regulator. Oral PG infection promotes atherosclerosis and induces significant metabolic changes, including reduced serum HDL and reduced hepatic SR-B1 and ABCA1 expression, as well as changes in intestinal microbiota. Our study suggests that intestinal dysbiosis accompanies periodontitis and could play a role in atherosclerosis.


Lymph node metastasis in breast cancer xenografts is associated with increased regions of extravascular drain, lymphatic vessel area, and invasive phenotype.
Interactions between the tumor stromal compartment and cancer cells play an important role in the spread of cancer. In this study, we have used noninvasive in vivo magnetic resonance imaging (MRI) of two human breast cancer models with significantly different invasiveness, to quantify and understand the role of interstitial fluid transport, lymphatic-convective drain, and vascularization in the regional spread of breast cancer to the axillary lymph nodes. Quantitative fluorescence microscopy was done to morphometrically characterize lymphatic vessels in these tumors. Significant differences in vascular and extravascular transport variables as well as in lymphatic vessel morphology were detected between the two breast cancer models, which also exhibited significant differences in lymph node and lung metastasis. These data are consistent with a role of lymphatic drain in lymph node metastasis and suggest that increased lymph node metastasis may occur due to a combination of increased invasiveness, and reduced extracellular matrix integrity allowing increased pathways of least resistance for the transport of extravascular fluid, as well as tumor cells. It is also possible that lymph node metastasis occurred via the cancer cell-bearing tumoral lymphatic vessels. The congestion of these tumoral lymphatics with cancer cells may have restricted the entry and transport of macromolecules.


Incidence of new-onset and flare of preexisting psoriasis during rituximab therapy for rheumatoid arthritis: data from the French AIR registry.
Psoriasis could be a paradoxical reaction to tumor necrosis factor-α antagonist therapy and it has been reported with rituximab therapy. Our objective was to assess the rates of new-onset and flare of preexisting psoriasis in patients taking rituximab for rheumatoid arthritis (RA). The nationwide multicenter prospective AutoImmunity and Rituximab (AIR) registry was set up in 2006 by the French Society for Rheumatology to collect data on patients taking rituximab for joint diseases. We identified patients with RA in the registry who had psoriasis listed as an adverse drug reaction, and we obtained additional information from their physicians if needed. We computed the incidence rates of new-onset and flare of preexisting psoriasis according to the rituximab exposure time. Among the 1927 patients in the registry with RA, 2 had new-onset and 5 had flare of preexisting psoriasis after a median followup of 39.2 weeks. Incidence rates were 1.04/1000 person-years (95% CI 0.13 to 3.8) for new-onset psoriasis and 2.6/1000 person-years (95% CI 0.84 to 6.1) for flare of preexisting psoriasis. Rituximab rechallenge in the 2 new-onset cases and in 2 flare cases was not followed by recurrence or exacerbation of psoriasis. Two of the 5 flare cases developed after discontinuation of methotrexate. Despite the small number of cases observed, leading to wide CI, the incidence rates in our study do not support a causative role of rituximab therapy in new-onset or flare of preexisting psoriasis in patients with RA.


Controlling for caffeine in cardiovascular research: A critical review.
Caffeine, the most widely consumed drug in the world, exerts numerous effects on cardiovascular activity. Thus, it is important and advisable to control for caffeine consumption in studies examining caffeine and/or cardiovascular activity and reactivity. This paper 1) reviews the literature concerning caffeine's effects on cardiovascular parameters; 2) summarizes the widely varying protocols used to control for the drug in extant cardiovascular literature, and 3) provide guidelines for caffeine control procedures to minimize potentially confounding acute and withdrawal effects of the drug. An abstention period equal to the average half-life of the drug is recommended for creation of methodological controls for caffeine. Additional methodological recommendations are described concerning factors that moderate the half-life of caffeine. When feasible, researchers should consider and aim to control for caffeine's acute and extended psychophysiological effects. This understudied issue has fundamental implications for caffeine-related investigations and research in psychophysiology and behavioral medicine.


Effects of corticosteroids on bilirubin metabolism in patients with Gilbert's syndrome.
To elucidate the mechanism whereby corticosteroids decrease the serum bilirubin concentration, changes in bilirubin metabolism were studied in patients with Gilbert's syndrome using a bilirubin load test and/or nicotinic acid test before and after corticosteroid treatment. Steroid administration increased hepatic clearance and uptake of bilirubin; the transfer rate for biliary excretion was unaffected. These results suggest that the main effect was enhancement of hepatic uptake or storage of bilirubin, which may be an important mechanism whereby corticosteroids reduce serum bilirubin concentrations in various liver disorders.


A probabilistic map of the human ventral sensorimotor cortex using electrical stimulation.
The human ventral sensorimotor cortex (vSMC) is involved in facial expression, mastication, and swallowing, as well as the dynamic and highly coordinated movements of human speech production. However, vSMC organization remains poorly understood, and previously published population-driven maps of its somatotopy do not accurately reflect the variability across individuals in a quantitative, probabilistic fashion. The goal of this study was to describe the responses to electrical stimulation of the vSMC, generate probabilistic maps of function in the vSMC, and quantify the variability across individuals. Photographic, video, and stereotactic MRI data of intraoperative electrical stimulation of the vSMC were collected for 33 patients undergoing awake craniotomy. Stimulation sites were converted to a 2D coordinate system based on anatomical landmarks. Motor, sensory, and speech stimulation responses were reviewed and classified. Probabilistic maps of stimulation responses were generated, and spatial variance was quantified. In 33 patients, the authors identified 194 motor, 212 sensory, 61 speech-arrest, and 27 mixed responses. Responses were complex, stereotyped, and mostly nonphysiological movements, involving hand, orofacial, and laryngeal musculature. Within individuals, the presence of oral movement representations varied; however, the dorsal-ventral order was always preserved. The most robust motor responses were jaw (probability 0.85), tongue (0.64), lips (0.58), and throat (0.52). Vocalizations were seen in 6 patients (0.18), more dorsally near lip and dorsal throat areas. Sensory responses were spatially dispersed; however, patients' subjective reports were highly precise in localization within the mouth. The most robust responses included tongue (0.82) and lips (0.42). The probability of speech arrest was 0.85, highest 15-20 mm anterior to the central sulcus and just dorsal to the sylvian fissure, in the anterior precentral gyrus or pars opercularis. The authors report probabilistic maps of function in the human vSMC based on intraoperative cortical electrical stimulation. These results define the expected range of mapping outcomes in the vSMC of a single individual and shed light on the functional organization of the vSMC supporting speech motor control and nonspeech functions.


Bleomycin injury of the lung in a mast-cell-deficient model.
Lung mast cell hyperplasia and fibrosis is induced by bleomycin lung injury. The role of the mast cell in this process of injury and resultant fibrosis is unclear. Mutant mi/mi mice, profoundly mast-cell-deficient, were treated with intraperitoneal bleomycin and demonstrated minimal acute inflammatory and chronic fibrotic responses. Lung histamine values determined at 14 and 42 days after bleomycin injury in mi/mi mice were not increased compared to untreated mi/mi animals. However, lung histamine levels in normal mice demonstrated a 300% increase over controls on Day 14 after bleomycin injury, and then returned to baseline by Day 42. The mi/mi BAL cell recovery at 2 weeks after injury and lung hydroxyproline levels at 4 weeks after injury were not altered from baseline. The normal litter mates, in contrast, demonstrated significant increases compared to controls in both of these parameters (p < 0.01, p < 0.04). Although the mi/mi mouse is also deficient in basophils, natural killer cells and functional osteoclasts, there is no evidence of lowered pulmonary defense mechanism and neutrophils and alveolar macrophages are present in normal numbers. This investigation supports the hypothesis that the mast cell contributes to bleomycin-induced lung injury and fibrosis.


Large volume transfusion with whole blood is safe compared with component therapy.
Transfusion with uncrossmatched cold-stored low-titer group O-positive or -negative whole blood (WB) in civilian trauma has been investigated as an alternative to component therapy but only in limited volumes. To our knowledge, this is the first analysis of the safety and efficacy of large volume transfusion of patients with trauma with WB. This is a retrospective cohort analysis comparing trauma patients resuscitated with component therapy (COMP) versus component therapy plus WB. The COMP group was comprised of patients who presented from January 2017 through June 2018 and the WB group from patients who presented from July 2018 through January 2019 after WB became available. We included patients if they received 1 unit of WB or red blood cells (RBCs) within 24 hours of admission and had massive transfusion protocol activated. We used bivariate analysis to compare groups. For analysis, one unit of WB equaled 1 unit of RBCs, 1 unit of plasma, and 1/6 of a unit of platelets. Forty-two patients received WB and 83 patients received COMP with similar baseline characteristics. Patients had a median age of 41 years (interquartile range [IQR], 28-61 years) and 73% were male. Thirty percent had penetrating injuries with a median Injury Severity Score of 29 (IQR, 17-38). The WB group received a median of 6.5 units (IQR, 3-11). The WB group received significantly more component-equivalent units but with a plasma/RBC ratio of 0.94:1 compared with 0.8:1 (p < 0.001). There were no differences in 24-hour mortality (COMP, 27% vs. WB, 29%, p = 0.8) or 30-day mortality (COMP, 46% vs. WB, 58% p = 0.2). There were no transfusion reactions. Transfusion utilizing primarily WB in civilian trauma is feasible, even in large volumes. It appears to be a safe and effective addition to component therapy and may lead to a more balanced resuscitation but with more overall product used. Therapeutic study, Level IV.


Expression of wild-type and nuclear localization-deficient human lamin A in chick myogenic cells.
Previous analysis of chick embryonic muscle (CEM) differentiation in vivo and in ovo demonstrated that lamin A accumulation to steady-state levels preceded the accumulation of muscle-specific proteins. These observations have suggested the appearance of A-type lamins may be important for differentiation. To test this hypothesis, we have temporally and quantitatively altered the expression of A-type lamins in CEM cells by transient transfection of wild-type (wt; pHLA) or nuclear localization-deficient (NLd; pHLA-del) human lamin A expression plasmids. Transfected CEM cells synthesized the wt and NLd human lamin As to high levels, both of which were resistant to high-salt extraction. The wt human lamin A localized to the nucleus, whereas the NLd protein showed cytoplasmic staining patterns, as well as time-dependent nuclear localization. The presence of endogenous chicken lamins A and B2 in NLd human lamin A cytoplasmic structures suggested the interspecies lamin copolymerization. Thus, this approach may provide a possible method for analysis of lamin-lamin or lamin-lamina component interactions in vivo. With regard to muscle differentiation, CEM cells transfected with either pHLA or pHLA-del demonstrated moderate and transient increased levels of the muscle-specific myosin heavy chain and creatine kinase activity. These increases appeared temporally and quantitatively to reflect the transient accumulation of the human lamin As. In contrast, beta-tubulin and skeletal tropomyosin protein accumulations appeared unaffected. On the basis of these results, we suggest that nuclear lamina content and structure may play a limited, permissive and indirect role in the temporally regulated expression of the myogenic phenotype.


INFECTIOUS CATARRH OF THE ALBINO RAT : II. THE CAUSAL RELATION OF COCCOBACILLIFORM BODIES.
Evidence is presented that infectious catarrh of the rat is caused by Gram-negative coccobacilliform bodies which resemble the agents of fowl coryza and mouse catarrh. These bodies were demonstrable microscopically in at least 90 per cent of the exudates from the nasal passages and middle ears. The characteristic syndrome was reproduced in selected rats by the nasal instillation of pure tissue culture growths and was maintained for 10 successive passages by the injection of exudate. The experimentally produced disease was also communicable by direct contact. The biological characteristics of the specific bodies are discussed in relation to the organisms of the pleuropneumonia group.


Ultrathin graphdiyne film on graphene through solution-phase van der Waals epitaxy.
Graphdiyne (GDY) is an ordered two-dimensional (2D) carbon allotrope comprising sp- and sp2-hybridized carbon atoms with high degrees of π-conjugation, which features a natural band gap and superior electric properties. However, the synthesis of one- or few-layer GDY remains challenging because of the free rotation around alkyne-aryl single bonds and the lack of thickness control. We report the facile synthesis of an ultrathin single-crystalline GDY film on graphene through a solution-phase van der Waals epitaxial strategy. The weak admolecule-substrate interaction at the heterojunction drastically relaxes the large lattice mismatch between GDY and graphene. It allows the fast in-plane coupling of admolecules and slow out-of-plane growth toward the formation of an incommensurately stacked heterostructure, which is composed of single-layer graphene and few-layer ABC-stacked GDY, as directly observed by electron microscopy and identified from Raman fingerprints. This study provides a general route not only to the bottom-up synthesis of intriguing 2D acetylenic carbon allotropes but also to the device fabrication for the direct measurement of their intrinsic electrical, mechanical, and thermal properties.


Bridging the gap of axonal regeneration in the central nervous system: A state of the art review on central axonal regeneration.
Neuronal regeneration in the central nervous system (CNS) is an important field of research with relevance to all types of neuronal injuries, including neurodegenerative diseases. The glial scar is a result of the astrocyte response to CNS injury. It is made up of many components creating a complex environment in which astrocytes play various key roles. The glial scar is heterogeneous, diverse and its composition depends upon the injury type and location. The heterogeneity of the glial scar observed in different situations of CNS damage and the consequent implications for axon regeneration have not been reviewed in depth. The gap in this knowledge will be addressed in this review which will also focus on our current understanding of central axonal regeneration and the molecular mechanisms involved. The multifactorial context of CNS regeneration is discussed, and we review newly identified roles for components previously thought to solely play an inhibitory role in central regeneration: astrocytes and p75NTR and discuss their potential and relevance for deciding therapeutic interventions. The article ends with a comprehensive review of promising new therapeutic targets identified for axonal regeneration in CNS and a discussion of novel ways of looking at therapeutic interventions for several brain diseases and injuries.


Investigation of an Indoor Air Quality Sensor for Asthma Management in Children.
Monitoring indoor air quality is critical because Americans spend 93% of their life indoors, and around 6.3 million children suffer from asthma. We want to passively and unobtrusively monitor the asthma patient's environment to detect the presence of two asthma-exacerbating activities: smoking and cooking using the Foobot sensor. We propose a data-driven approach to develop a continuous monitoring-activity detection system aimed at understanding and improving indoor air quality in asthma management. In this study, we were successfully able to detect a high concentration of particulate matter, volatile organic compounds, and carbon dioxide during cooking and smoking activities. We detected 1) smoking with an error rate of 1%; 2) cooking with an error rate of 11%; and 3) obtained an overall 95.7% percent accuracy classification across all events (control, cooking and smoking). Such a system will allow doctors and clinicians to correlate potential asthma symptoms and exacerbation reports from patients with environmental factors without having to personally be present.