Increased apoptotic cells in bone marrow biopsies from patients with aplastic anaemia.
In order to investigate the involvement of apoptosis in the pathogenesis of aplastic anaemia (AA) we determined the proportion of apoptotic cells in paraffin-embedded bone marrow biopsies from patients with aplastic anaemia using an in situ TdT-catalysed DNA nick end labelling (TUNEL) staining method. A significant increase in the proportion of mononuclear apoptotic cells was demonstrated in biopsies from patients with aplastic anaemia (8.19 +/- 1.45%) when compared with controls (2.07 +/- 0.86%). These data support the view that apoptosis may play a role in the pathophysiology of bone marrow failure.


Construction of a functional silk-based biomaterial complex with immortalized chondrocytes in vivo.
To explore the feasibility of constructing a functional biomaterial complex with regenerated silk fibroin membrane and immortalized chondrocytes in vivo. Rat auricular chondrocytes (RACs) were transfected with the lentivirus vector pGC-FU-hTERT-3FLAG or pGC-FU-GFP-3FLAG, encoding the human telomerase reverse transcriptase (hTERT) or GFP gene. The effects of regenerated silk fibroin film on the adhesion, growth of immortalized chondrocytes and expression of collagen II in vitro were analyzed with immunofluorescent histochemistry. Immortalized RACs were transformed. Induction by nutrient medium promoted higher expression levels of collagen II in transformed chondrocytes. The regenerated silk fibroin film was not cytotoxic to immortalized chondrocytes and had no adverse influence on their adhesion. Collagen II expression was good in the immortalized chondrocytes in vivo. The construction of a silk-based biomaterial complex with immortalized chondrocytes may provide a feasible kind of functional biomaterial for the repair of cartilage defects in clinical applications.


Parametric formulation of the dielectric function of palladium and palladium hydride thin films.
A parametric description of the dielectric function of Pd thin films with thicknesses between 10 and 30 nm is reported. These films were grown at room temperature on amorphous quartz substrates by electron beam evaporation, with a base pressure of 7.0×10(-7) mbar. By using nonpolarized normal incident light, transmission spectra were measured for wavelengths between 240 and 1050 nm. Inversion of the spectra by means of a projected gradient method enables us to obtain the mean dielectric function of the Pd grains in the films. We follow the Brendel-Bormann model to describe the frequency dependence of the dielectric function, with the plasma frequency, collision frequency, and screening factor as parameters in the free electron term. The contributions of bound electrons and their interband transitions, described in terms of Lorentz oscillators, involve the resonance frequencies, decay times, strengths, and Gaussian widths as parameters of the model. All these parameters have been optimized from the Pd grains' dielectric function, which fits the transmission spectra. A similar procedure was followed for Pd films exposed to a hydrogen atmosphere close to one bar. Thus, the dielectric functions of palladium and palladium hydride can easily be calculated through spectral ranges covering near-ultraviolet, visible, and near-infrared wavelengths. This can be used to model the behavior of nano-sized structures in which palladium particles or thin films are exposed to hydrogen pressures close to one bar.


Archaea in Natural and Impacted Brazilian Environments.
In recent years, archaeal diversity surveys have received increasing attention. Brazil is a country known for its natural diversity and variety of biomes, which makes it an interesting sampling site for such studies. However, archaeal communities in natural and impacted Brazilian environments have only recently been investigated. In this review, based on a search on the PubMed database on the last week of April 2016, we present and discuss the results obtained in the 51 studies retrieved, focusing on archaeal communities in water, sediments, and soils of different Brazilian environments. We concluded that, in spite of its vast territory and biomes, the number of publications focusing on archaeal detection and/or characterization in Brazil is still incipient, indicating that these environments still represent a great potential to be explored.


The impact of endurance exercise on global and AMPK gene-specific DNA methylation.
Alterations in gene expression as a consequence of physical exercise are frequently described. The mechanism of these regulations might depend on epigenetic changes in global or gene-specific DNA methylation levels. The AMP-activated protein kinase (AMPK) plays a key role in maintenance of energy homeostasis and is activated by increases in the AMP/ATP ratio as occurring in skeletal muscles after sporting activity. To analyze whether exercise has an impact on the methylation status of the AMPK promoter, we determined the AMPK methylation status in human blood samples from patients before and after sporting activity in the context of rehabilitation as well as in skeletal muscles of trained and untrained mice. Further, we examined long interspersed nuclear element 1 (LINE-1) as indicator of global DNA methylation changes. Our results revealed that light sporting activity in mice and humans does not alter global DNA methylation but has an effect on methylation of specific CpG sites in the AMPKα2 gene. These regulations were associated with a reduced AMPKα2 mRNA and protein expression in muscle tissue, pointing at a contribution of the methylation status to AMPK expression. Taken together, these results suggest that exercise influences AMPKα2 gene methylation in human blood and eminently in the skeletal muscle of mice and therefore might repress AMPKα2 gene expression.


Concordance in Breast Cancer Grading by Artificial Intelligence on Whole Slide Images Compares With a Multi-Institutional Cohort of Breast Pathologists.
Breast carcinoma grade, as determined by the Nottingham Grading System (NGS), is an important criterion for determining prognosis. The NGS is based on 3 parameters: tubule formation (TF), nuclear pleomorphism (NP), and mitotic count (MC). The advent of digital pathology and artificial intelligence (AI) have increased interest in virtual microscopy using digital whole slide imaging (WSI) more broadly. To compare concordance in breast carcinoma grading between AI and a multi-institutional group of breast pathologists using digital WSI. We have developed an automated NGS framework using deep learning. Six pathologists and AI independently reviewed a digitally scanned slide from 137 invasive carcinomas and assigned a grade based on scoring of the TF, NP, and MC. Interobserver agreement for the pathologists and AI for overall grade was moderate (κ = 0.471). Agreement was good (κ = 0.681), moderate (κ = 0.442), and fair (κ = 0.368) for grades 1, 3, and 2, respectively. Observer pair concordance for AI and individual pathologists ranged from fair to good (κ = 0.313-0.606). Perfect agreement was observed in 25 cases (27.4%). Interobserver agreement for the individual components was best for TF (κ = 0.471 each) followed by NP (κ = 0.342) and was worst for MC (κ = 0.233). There were no observed differences in concordance amongst pathologists alone versus pathologists + AI. Ours is the first study comparing concordance in breast carcinoma grading between a multi-institutional group of pathologists using virtual microscopy to a newly developed WSI AI methodology. Using explainable methods, AI demonstrated similar concordance to pathologists alone.


Induction of capacitation in human spermatozoa in vitro by an endometrial sialic acid-binding protein.
A Ca(2+)-dependent sialic acid-binding protein (SABP) of human endometrium, which specifically bound to human sperm head plasma membrane in vitro, was found to increase the percentage motility and acrosome-reacted pattern of uncapacitated spermatozoa. The protein was synthesized in the endometrium and secreted into the uterine fluid. This intra-uterine factor, which is apparently advantageous in vitro in inducing human sperm capacitation, may play a significant role in promoting the post-release maturation of ejaculated spermatozoa by enhancing 45Ca uptake into spermatozoa by a pathway which is insensitive to calcium-channel blockers. However, the 45Ca uptake could be enhanced on exposure to the divalent cation ionophore A23187 and inhibited in the presence of the calmodulin inhibitor trifluoperazine. The SABP also induces an increase in intracellular Ca2+ in spermatozoa, as seen by FURA-2 AM studies. Furthermore, overlay studies show human SABP to be a Ca(2+)-binding protein. The data presented here suggest that SABP induces in-vitro sperm capacitation and the subsequent acrosome reaction by increasing intracellular Ca2+ concentration.


Atypical Mycobacterial Infections of the Upper Extremity.
Atypical mycobacterial infections of upper extremity synovial-lined structures are often misdiagnosed and unrecognized. Despite an increasing incidence, lack of physician awareness of these pathogens may result in considerable delay in diagnosis and management, potentially leading to permanent disability. The authors conducted a literature review and analyzed 31 cases of penetrating atypical mycobacterial infection to better understand the clinical characteristics and to evaluate their posttreatment complication rate compared with available literature. Medical records for culture-positive cases of tenosynovial or intra-articular atypical mycobacterial infections of the upper extremity that were treated were retrospectively reviewed. Treatment outcomes were analyzed against published case reviews and case series. Thirty-one cases of penetrating atypical mycobacterial infection were identified. Mycobacterium marinum (n=11) was the most common organism and was associated with aquatic exposure. Twenty-eight cases received empiric treatment, 17 of which received contraindicated treatment. Patients saw an average of 5 physicians prior to receiving an accurate diagnosis, and the mean time to diagnosis was 10 months. All cases received antibiotic treatment in addition to surgical management. Twenty cases (68%) failed treatment outcomes. Delay in diagnosis and inappropriate management of atypical mycobacterial infections may lead to a treatment failure rate that is higher than what has been reported in the literature. Mycobacterium avium and M fortuitum had significantly higher failure rates than other organisms. A high index of suspicion is required to make a diagnosis and prevent residual disability. [Orthopedics. 2018; 41(3):e383-e388.].


Muscle Transcriptome Analysis Reveals Potential Candidate Genes and Pathways Affecting Intramuscular Fat Content in Pigs.
Intramuscular fat (IMF) content plays an essential role in meat quality. For identifying potential candidate genes and pathways regulating IMF content, the IMF content and the longissimus dorsi transcriptomes of 28 purebred Duroc pigs were measured. As a result, the transcriptome analysis of four high- and four low-IMF individuals revealed a total of 309 differentially expressed genes (DEGs) using edgeR and DESeq2 (p < 0.05, |log2(fold change)| ≥ 1). Functional enrichment analysis of the DEGs revealed 19 hub genes significantly enriched in the Gene Ontology (GO) terms and pathways (q < 0.05) related to lipid metabolism and fat cell differentiation. The weighted gene coexpression network analysis (WGCNA) of the 28 pigs identified the most relevant module with 43 hub genes. The combined results of DEGs, WGCNA, and protein-protein interactions revealed ADIPOQ, PPARG, LIPE, CIDEC, PLIN1, CIDEA, and FABP4 to be potential candidate genes affecting IMF. Furthermore, the regulation of lipolysis in adipocytes and the peroxisome proliferator-activated receptor (PPAR) signaling pathway were significantly enriched for both the DEGs and genes in the most relevant module. Some DEGs and pathways detected in our study play essential roles and are potential candidate genes and pathways that affect IMF content in pigs. This study provides crucial information for understanding the molecular mechanism of IMF content and would be helpful in improving pork quality.


Genome-wide identification and characterization of TIFY family genes in Moso Bamboo (Phyllostachys edulis) and expression profiling analysis under dehydration and cold stresses.
The proteins containing the TIFY domain belong to a plant-specific family of putative transcription factors and could be divided into four subfamilies: ZML, TIFY, PPD and JAZ. They not only function as key regulators of jasmonate hormonal response, but are also involved in responding to abiotic stress. In this study, we identified 24 TIFY genes (PeTIFYs) in Moso bamboo (Phyllostachys edulis) of Poaceae by analyzing the whole genome sequence. One PeTIFY belongs to TIFY subfamily, 18 and five belong to JAZ and ZML subfamilies, respectively. Two equivocal gene models were re-predicted and a putative retrotransposition event was found in a ZML protein. The distribution and conservation of domain or motif, and gene structure were also analyzed. Phylogenetic analysis with TIFY proteins of Arabidopsis and Oryza sativa indicated that JAZ subfamily could be further divided to four groups. Evolutionary analysis revealed intragenomic duplication and orthologous relationship between P. edulis, O. sativa, and B. distachyon. Calculation of the non-synonymous (Ka) and synonymous (Ks) substitution rates and their ratios indicated that the duplication of PeTIFY may have occurred around 16.7 million years ago (MYA), the divergence time of TIFY family among the P. edulis-O. sativa, P. edulis-B. distachyon, and O. sativa-B. distachyon was approximately 39 MYA, 39 MYA, and 45 MYA, respectively. They appear to have undergone extensive purifying selection during evolution. Transcriptome sequencing revealed that more than 50% of PeTIFY genes could be up-regulated by cold and dehydration stresses, and some PeTIFYs also share homology to know TIFYs involved in abiotic stress tolerance. Our results made insights into TIFY family of Moso bamboo, an economically important non-timber forest resource, and provided candidates for further identification of genes involved in regulating responses to abiotic stress.


Crystal structure of the complex of the variable domain of antibody D1.3 and turkey egg white lysozyme: a novel conformational change in antibody CDR-L3 selects for antigen.
The crystal structure of the Fv fragment of the murine monoclonal anti-lysozyme antibody D1.3, complexed with turkey egg-white lysozyme (TEL), is presented. D1.3 (IgG1, kappa) is a secondary response antibody specific for hen egg-white lysozyme (HEL). TEL and HEL are homologous and differ in amino acid sequence in the antibody-antigen interface only at position 121. The side-chain of HEL residue Gln121 makes a pair of hydrogen bonds to main-chain atoms of the antibody light chain. In the D1.3-TEL structure, TEL residue His121 makes only one hydrogen bond with the light chain as a result of 129 degree and 145 degree change in peptide torsion angles for residues Trp92 and Ser93. Probably as a consequence of this conformational change, the D1.3-TEL association occurs at a much slower rate than the D1.3-HEL association. The D1.3-TEL complex is destabilized with respect to the D1.3-HEL interaction by the loss of two hydrogen bonds, exclusively due to the substitution of histidine for glutamine. While antibodies of secondary responses are indeed highly specific for antigen, this work demonstrates that by undergoing subtle conformational change antibodies can still recognize mutated protein antigens, albeit at a cost to affinity.


Quality control of DXA instruments in multicenter trials.
Dual-energy X-ray absorptiometry (DXA) has become the measurement of choice for multicenter trials with bone density endpoints. When performing DXA measurements with several different systems, it is important to implement a quality assurance program to guarantee that any observed density changes are real and not due to machine and/or operator variability. In this study, we present a series of procedures based on phantom measurements designed to monitor DXA instrument stability. Techniques for longitudinal evaluation of machine performance and cross-calibration of instruments are described. These procedures are then demonstrated using quality assurance data collected from a number of different DXA scanners. Together these methods provide a defined approach to instrument quality control. Though based primarily on the use of spinal phantoms, these procedures can be generalized for use in any multicenter DXA study.


Four novel MSH2 / MLH1 gene mutations in portuguese HNPCC families.
Hereditary non-polyposis colorectal cancer (HNPCC) is considered to be determined by germline mutations in the mismatch repair (MMR) genes, especially MSH2 and MLH1. While screening for mutations in these two genes in HNPCC portuguese families, 3 previously unreported MSH2 and 1 MLH1 mutations have been identified in families meeting strict Amsterdam criteria. Hum Mutat 15:116, 2000.


PI3K/Akt/mTOR signaling pathway in cancer stem cells: from basic research to clinical application.
Cancer stem cells (CSCs) are a subpopulation of tumor cells that possess unique self-renewal activity and mediate tumor initiation and propagation. The PI3K/Akt/mTOR signaling pathway can be considered as a master regulator for cancer. More and more recent studies have shown the links between PI3K/Akt/mTOR signaling pathway and CSC biology. Herein, we provide a comprehensive review on the role of signaling components upstream and downstream of PI3K/Akt/mTOR signaling in CSC. In addition, we also summarize various classes of small molecule inhibitors of PI3K/Akt/mTOR signaling pathway and their clinical potential in CSC. Overall, the current available data suggest that the PI3K/Akt/mTOR signaling pathway could be a promising target for development of CSC-target drugs.


That 'gut feeling'.
Upper abdominal pain can be a dilemma in general practice. Sometimes the likely diagnosis is clear in your mind after the first sentence of the patient's history is revealed--and only gets confirmed the further you delve into the history and examination. On other occasions the diagnosis is far more elusive. In this issue of Australian Family Physician, Sprio Tsipouras raises the importance of the doctor's 'gestalt' in managing these elusive cases. What is gestalt? How do we develop it?


Insulin stimulates the phosphorylation and activity of farnesyltransferase via the Ras-mitogen-activated protein kinase pathway.
Farnesylation of p21Ras by farnesyltransferase (FTase) is obligatory for anchoring p21Ras to the plasma membrane, where it can be activated by growth factors. Insulin significantly stimulates the phosphorylation of the alpha-subunit of FTase (4-fold) and the enzymatic activity of FTase in 3T3-L1 fibroblasts and adipocytes. FTase activity was assessed by the amount of [3H] mevalonate (a precursor of farnesyl) incorporated into p21Ras in vivo and by quantitating the amount of farnesylated p21Ras before and after insulin administration. Insulin-stimulated phosphorylation of the alpha-subunit of FTase in 3T3-L1 fibroblasts and adipocytes was blocked by the mitogen-activated protein/extracellular-signal regulated kinase-kinase inhibitor, PD98059, but not by wortmannin or bisindolylmaleimide. Additionally, PD98059 blocked insulin-stimulated [3H]mevalonic incorporation and farnesylation of unprocessed p21Ras in both cell lines. Furthermore, expression of the dominant negative mutant of p21Ras precluded insulin-stimulated phosphorylation of the FTase alpha-subunit and activation of its enzymatic activity. In contrast, 3T3-L1 fibroblasts, expressing the constitutively active Raf-1, exhibited enhanced phosphorylation of the FTase alpha-subunit. It seems that insulin's effect on the phosphorylation and activation of FTase in both fibroblasts and adipocytes is mediated via the Ras pathway, resulting in a positive feedback augmentation of the cellular pool of farnesylated p21Ras.


Fatalities temporally associated with the ingestion of ibogaine.
Ibogaine is a naturally occurring psychoactive plant alkaloid that is used globally in medical and nonmedical settings for opioid detoxification and other substance use indications. All available autopsy, toxicological, and investigative reports were systematically reviewed for the consecutive series of all known fatalities outside of West Central Africa temporally related to the use of ibogaine from 1990 through 2008. Nineteen individuals (15 men, four women between 24 and 54 years old) are known to have died within 1.5-76 h of taking ibogaine. The clinical and postmortem evidence did not suggest a characteristic syndrome of neurotoxicity. Advanced preexisting medical comorbidities, which were mainly cardiovascular, and/or one or more commonly abused substances explained or contributed to the death in 12 of the 14 cases for which adequate postmortem data were available. Other apparent risk factors include seizures associated with withdrawal from alcohol and benzodiazepines and the uninformed use of ethnopharmacological forms of ibogaine.


Severe recalcitrant cutaneous manifestations in systemic lupus erythematosus successfully treated with fumaric acid esters.
Fumaric acid esters (FAEs) have proven efficacy in the treatment of psoriasis and have been in use for decades. More recently, as their mechanism of action and abundant immunomodulatory effects become clearer, the potential benefits of treating other inflammatory skin conditions using FAEs are increasingly being recognized. The use of FAEs as combination systemic therapy has not been well studied and data are lacking regarding the safety and efficacy of this type of therapy. In this case report, three patients with severe, extensive and recalcitrant cutaneous manifestations of systemic lupus erythematosus (SLE) (one case of disseminated discoid lesions and two with severe chilblain lesions) were treated with Fumaderm® (containing the FAE dimethylfumarate and monoethylhydrogen fumarate salts), after failing to respond to a multitude of other monotherapies and combination therapies. All patients showed a substantial clinical response when FAEs were added to their treatment, with concurrent improvements in quality-of-life instrument scores. The treatment was well tolerated in the context of systemic organ involvement and as combination therapy with other agents, such as hydroxychloroquine and mycophenolate mofetil. These cases of SLE illustrate the potential use of FAEs in severe, disfiguring and otherwise therapy-resistant skin lesions, including, to our knowledge, the first two reported cases of FAE-treated chilblain lupus erythematosus.


Neurospheres: insights into neural stem cell biology.
Neural stem cells (NSC) are a tissue-specific subtype of self-renewing and multipotent cells that can give rise to all neural populations. In this review, the importance of maintaining cell-cell contacts in the study of NSC is highlighted, and data obtained from some crucial single-cell studies is compared to results obtained from neurospheres, where aggregates of NSC are grown in suspension. In particular, results that indicate how this culture system may be well suited to analyze NSC plasticity, cell-cell, and cell-extracellular matrix (ECM) interactions are pointed out, and the hypothesis that cell-cell and cell-ECM contacts may be essential for NSC maintenance, survival, and proliferation is highlighted. Finally, it is suggested that neurospheres might play a role in the study of context-dependent behavior of NSC in niches by providing a system where NSC can be challenged chemically or biologically and analyzed in vitro, in a time- and context-dependent manner.


High-relaxivity magnetic resonance imaging contrast agents. Part 2. Optimization of inner- and second-sphere relaxivity.
The observed relaxivity of gadolinium-based contrast agents has contributions from the water molecule(s) that bind directly to the gadolinium ion (inner-sphere water), long-lived water molecules and exchangeable protons that make up the second-sphere of coordination, and water molecules that diffuse near the contrast agent (outer-sphere). Inner- and second-sphere relaxivity can both be increased by optimization of the lifetimes of the water molecules and protons in these coordination spheres, the rotational motion of the complex, and the electronic relaxation of the gadolinium ion. We sought to identify new high-relaxivity contrast agents by systematically varying the donor atoms that bind directly to gadolinium to increase inner-sphere relaxivity and concurrently including substituents that influence the second-sphere relaxivity. Twenty gadolinium-1,4,7,10-tetraazacyclo-dodecane-N,N',N″,N'″-tetraacetato derivatives were prepared and their relaxivity determined in presence and absence of human serum albumin as a function of temperature and magnetic field. Data was analyzed to extract the underlying molecular parameters influencing relaxivity. Each compound had a common albumin-binding group and an inner-sphere donor set comprising the 4 tertiary amine N atoms from cyclen, an α-substituted acetate oxygen atom, 2 amide oxygen atoms, an inner-sphere water oxygen atom, and a variable donor group. Each amide nitrogen was substituted with different groups to promote hydrogen bonding with second-sphere water molecules. Relativities at 0.47 and 1.4 T, 37°C, in serum albumin ranged from 16.0 to 58.1 mM(-1)s(-1) and from 12.3 to 34.8 mM(-1)s(-1), respectively. The reduction of inner-sphere water exchange typical of amide donor groups could be offset by incorporating a phosphonate or phenolate oxygen atom donor in the first coordination sphere, resulting in higher relaxivity. Amide nitrogen substitution with pendant phosphonate or carboxylate groups increased relaxivity by as much as 88% compared with the N-methyl amide analog. Second-sphere relaxivity contributed as much as 24 and 14 mM(-1)s(-1) at 0.47 and 1.4 T, respectively. Water/proton exchange dynamics in the inner- and second-coordination sphere can be predictably tuned by choice of donor atoms and second-sphere substituents, resulting in high-relaxivity agents.


The energetics of CNS white matter.
The energetics of CNS white matter are poorly understood. We derive a signaling energy budget for the white matter (based on data from the rodent optic nerve and corpus callosum) which can be compared with previous energy budgets for the gray matter regions of the brain, perform a cost-benefit analysis of the energetics of myelination, and assess mechanisms for energy production and glucose supply in myelinated axons. We show that white matter synapses consume ≤0.5% of the energy of gray matter synapses and that this, rather than more energy-efficient action potentials, is the main reason why CNS white matter uses less energy than gray matter. Surprisingly, while the energetic cost of building myelin could be repaid within months by the reduced ATP cost of neuronal action potentials, the energetic cost of maintaining the oligodendrocyte resting potential usually outweighs the saving on action potentials. Thus, although it dramatically speeds action potential propagation, myelination need not save energy. Finally, we show that mitochondria in optic nerve axons could sustain measured firing rates with a plausible density of glucose transporters in the nodal membrane, without the need for energy transfer from oligodendrocytes.


Unfolded protein response suppresses cisplatin-induced apoptosis via autophagy regulation in human hepatocellular carcinoma cells.
It has been shown that drug resistance is extremely common in hepatocellular carcinoma (HCC) and is one of the major problems in HCC chemotherapy. However, the detailed mechanisms remain largely unknown. We have previously shown that endoplasmic reticulum (ER) stress is involved in the tumorigenesis of HCC. Here, we demonstrated that the unfolded protein response (UPR) inhibits cisplatin-induced HCC cell apoptosis. In HCC cells, cisplatin treatment triggers the UPR, which subsequently inhibits cisplatin-induced apoptosis. Importantly, mild ER stress precondition suppresses the sensitivity of HCC cells to cisplatin-induced apoptosis through autophagy regulation. Furthermore, heat-shock protein 27 (Hsp27) is involved in the cytoprotective role of the UPR in cisplatin-induced apoptosis. We also demonstrated that Hsp27 inhibits cisplatin- induced HCC cell death through autophagy activation. Taken together, our results indicate that the UPR inhibits cisplatin-induced apoptosis in HCC cells, at least in part, by Hsp27-mediated autophagy activation.


Effect of caldesmon on the position and myosin-induced movement of smooth muscle tropomyosin bound to actin.
It is known that the actin-binding protein caldesmon inhibits actomyosin ATPase activity and might in this way take part in the thin filament regulation of smooth muscle contraction. Although the molecular mechanism of this inhibition is unknown, it is clear that the presence of actin-bound tropomyosin is necessary for full inhibition. Recent evidence also suggests that the myosin-induced movement of tropomyosin plays a key role in regulation. In this work, fluorescence studies provide evidence to show that caldesmon interacts with and alters the position of tropomyosin in a reconstituted actin thin filament and thereby limits the ability of myosin heads to move tropomyosin. Caldesmon interacts with the Cys-190 region in the COOH-terminal half of tropomyosin, resulting in the movement of this part of tropomyosin to a new position on actin. Additionally, this constrains the myosin-induced movement of this region of tropomyosin. On the other hand, caldesmon does not appear to interact with the Cys-36 region in the NH2-terminal half of tropomyosin and neither alters the position of nor significantly constrains the myosin-induced movement of this part of tropomyosin. The ability of caldesmon to limit the myosin-induced movement of tropomyosin provides a possible molecular basis for the inhibitory function of caldesmon. The different movements of the two halves of tropomyosin indicate that actin-bound tropomyosin moves as a flexible molecule and not as a rigid rod. Interestingly, caldesmon, which inhibits tropomyosin's potentiation of actomyosin ATPase activity, moves tropomyosin in one direction, whereas myosin heads, which enhance potentiation, move tropomyosin in the opposite direction.


Effect of Fermented Medicinal Plants as Dietary Additives on Food Preference and Fecal Microbial Quality in Dogs.
This research determined the antioxidant activities of medicinal plants fermented by Enterococcus faecium and their subsequent applications as dog food additives. Turmeric (5%, w/v), glasswort (2.5%, w/v), Ganghwa mugwort (2.5%, w/v), and their mixture (5%, w/v) were fermented by autochthonous E. faecium (1%, v/v) for 72 h. Bacterial cell counts and pH were monitored during fermentation. Total polyphenol content (TPC), total flavonoid content (TFC), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, and intracellular superoxide scavenging activity in bovine mammary alveolar epithelial (MAC-T) cells were measured with the fermented and non-fermented samples. Only the antioxidant capacity of the mixture was increased after fermentation. However, intracellular superoxide level in MAC-T cells was significantly reduced after treatment with fermented plant samples (p < 0.001) as compared with that in non-fermented plants. Fermented plants were then sprayed at 1% (v/w) onto dog foods. TPC, TFC, ABTS radical scavenging activity, and DPPH radical scavenging activity of dog foods were significantly enhanced after the addition of fermented plants. Food preference testing was conducted using a two-pan method-control diet vs. four treatment diets-for 4 days for each additive diet, a total 16 days in 9 beagles. Feces were collected to enumerate bacterial counts. Preferences for glasswort and Ganghwa mugwort were higher than those of the control (p < 0.05). Furthermore, fecal microbiota enumeration displayed a higher number of beneficial microorganisms in treated groups. These results suggest that fermented plants with enhanced antioxidant abilities might be useful as potential additives for dog foods.


Shaping up synthetic cells.
How do the cells in our body reconfigure their shape to achieve complex tasks like migration and mitosis, yet maintain their shape in response to forces exerted by, for instance, blood flow and muscle action? Cell shape control is defined by a delicate mechanical balance between active force generation and passive material properties of the plasma membrane and the cytoskeleton. The cytoskeleton forms a space-spanning fibrous network comprising three subsystems: actin, microtubules and intermediate filaments. Bottom-up reconstitution of minimal synthetic cells where these cytoskeletal subsystems are encapsulated inside a lipid vesicle provides a powerful avenue to dissect the force balance that governs cell shape control. Although encapsulation is technically demanding, a steady stream of advances in this technique has made the reconstitution of shape-changing minimal cells increasingly feasible. In this topical review we provide a route-map of the recent advances in cytoskeletal encapsulation techniques and outline recent reports that demonstrate shape change phenomena in simple biomimetic vesicle systems. We end with an outlook toward the next steps required to achieve more complex shape changes with the ultimate aim of building a fully functional synthetic cell with the capability to autonomously grow, divide and move.


Mutations in the C-terminal part of insulin-like growth factor (IGF)-binding protein-1 result in dimer formation and loss of IGF binding capacity.
In an attempt to define domains in insulin-like growth factor (IGF)-binding protein-1 (IGFBP-1) that are involved in IGF binding, we subjected the carboxyl end of the coding region of IGFBP-1 cDNA to mutagenesis. Mutant cDNAs were isolated, characterized by sequencing, and cloned in an expression vector under control of the simian virus-40 (SV40) early promoter. The constructs were transfected into COS-1 cells, and the mutant proteins, secreted into the culture medium, were analyzed for IGF binding by ligand blotting. The results obtained show that deletion of the C-terminal 20 amino acids or introduction of frame-shifts in this region resulted in loss of IGF binding and for some mutants in the formation of dimeric IGFBP-1 molecules. These dimers are probably formed when cysteine-226 (Cys-226) is missing, and its putative partner is able to form intermolecular disulfide bonds. Site-directed mutagenesis demonstrated that most of the introduced point mutations in the C-terminal region did not affect IGF binding. Only mutation of Cys-226 to tyrosine completely abolished IGF binding, as did the introduction of a negatively charged amino acid in the vicinity of this residue. Again, dimers were observed, supporting that Cys-226 is essential for the conformation of IGFBP-1. In addition, our data suggest that an IGF-binding domain may be located in the vicinity of the intramolecular disulfide bond formed by Cys-226 and its putative partner.


Artificial intelligence and atrial fibrillation.
In the context of atrial fibrillation (AF), traditional clinical practices have thus fallen short in several domains, such as identifying patients at risk of incident AF or patients with concomitant undetected paroxysmal AF. Novel approaches leveraging artificial intelligence have the potential to provide new tools to deal with some of these old problems. To discuss the roles of artificial intelligence (AI)-enabled electrocardiogram (ECG) pertaining to AF, potential roles of deep learning (DL) models in the context of current knowledge gaps, as well as limitations of these models. An extensive search and review of the currently available literature on the topics. One key area where DL models can translate to better patient outcomes is through automated ECG interpretation. Challenges with regards to the benefits and harms of AF screening remain. In this context, a unique model was developed to detect underlying hidden AF from sinus rhythm. Knowledge gaps remain regarding the best ways to monitor patients with embolic stroke of undetermined source (ESUS) and identifying those who would benefit most from oral anticoagulation. The AI-enabled AF model is one potential way to tackle this complex problem as it could be used to identify a subset of high-risk ESUS patients likely to benefit from empirical oral anticoagulation. The role of DL models assessing AF burden from long-duration ECG data is also discussed as a way of guiding management. There is a trend towards the use of consumer-grade wristbands and watches to detect AF from photoplethysmography data. However, ECG currently remains the gold standard to detect arrythmias including AF. Lastly, the role of adequate external validation of the models and clinical trials to study true performance is discussed. Algorithms using AI to interpret ECGs in various new ways have been developed. While still, much work needs to be done, these technologies have shown enormous potential in a short span of time. With further advancements and continuous research, these novel ways of interpretation may well become part of everyday clinical workflow.


Effects of hydrolysed Saccharomyces cerevisiae yeast and yeast cell wall components on live performance, intestinal histo-morphology and humoral immune response of broilers.
1. The effects of enzymatically hydrolysed whole Saccharomyces cerevisiae yeast (HY) and the pellets of yeast cell wall (YCW) on production traits, the microbiology and histo-morphology of the small intestine, and humoral immune responses against Newcastle disease virus (NDV), of Ross 308 broilers were investigated. 2. The control group received a maize-soyabean meal based basal diet for 42 days. In the treated groups the basal diet was supplemented with 1 g/kg of HY and YCW. There were 8 replicate pens per group (n = 12 birds/pen). 3. HY and YCW supplementation improved live weight (P = 0·006) and FCR (P = 0·003) at 42-d as compared with the control group. 4. In the small intestine, Salmonella spp and Escherichia coli numbers were higher (P = 0·01) in the mucosa and lower (P = 0·01) in the digesta of the HY and the YCW fed groups at 25 d of age. Lactobacillus in the duodenal and jejunal digesta was higher (P < 0·05) in the HY and the YCW fed groups as compared with the control. 5. Following oral challenge with Salmonella pullorum, Escherichia coli and Lactobacillus increased (P < 0·05) in the mucosa and decreased in the digesta (P < 0·05) of the HY and YCW supplemented groups, relative to the control. 6. Supplementation of HY and YCW increased villus height in the jejunum (P = 0·02), width of villus in the ileum (P = 0·034) and number of goblet cells in villi of the jejunum (P = 0·006) and ileum (P = 0·01). 7. YCW increased antibody level against NDV at 21 and 42 d of age compared with the control and the HY supplemented diets (P < 0·05). 8. It was concluded that HY and YCW improved growth and feed efficiency in broilers, and considering the improvements in production traits and humoral immune responses, yeast cell wall may be a better dietary tool than the hydrolysed whole yeast cell as a performance enhancer for broilers.


All or nothing? Partial business shutdowns and COVID-19 fatality growth.
Incomplete vaccine uptake and limited vaccine availability for some segments of the population could lead policymakers to consider re-imposing restrictions to help reduce fatalities. Early in the pandemic, full business shutdowns were commonplace. Given this response, much of the literature on policy effectiveness has focused on full closures and their impact. But were complete closures necessary? Using a hand-collected database of partial business closures for all U.S. counties from March through December 2020, we examine the impact of capacity restrictions on COVID-19 fatality growth. For the restaurant and bar sector, we find that several combinations of partial capacity restrictions are as effective as full shutdowns. For example, point estimates indicate that, for the average county, limiting restaurants and bars to 25% of capacity reduces the fatality growth rate six weeks ahead by approximately 43%, while completely closing them reduces fatality growth by about 16%. The evidence is more mixed for the other sectors that we study. We find that full gym closures reduce the COVID-19 fatality growth rate, while partial closures may be counterproductive relative to leaving capacity unrestricted. Retail closures are ineffective, but 50% capacity limits reduce fatality growth. We find that restricting salons, other personal services and movie theaters is either ineffective or counterproductive.


Multimodal exploration of non-motor neural functions in ALS patients using simultaneous EEG-fNIRS recording.
Despite the high prevalence of non-motor impairments reported in patients with amyotrophic lateral sclerosis (ALS), little is known about the functional neural markers underlying such dysfunctions. In this study, a new dual-task multimodal framework relying on simultaneous electroencephalogram (EEG) and functional near-infrared spectroscopy (fNIRS) recordings was developed to characterize integrative non-motor neural functions in people with ALS. Simultaneous EEG-fNIRS data were recorded from six subjects with ALS and twelve healthy controls. Through a proposed visuo-mental paradigm, subjects performed a set of visuo-mental arithmetic operations. The data recorded were analyzed with respect to event-related changes both in the time and frequency domains for EEG and de/oxygen-hemoglobin level (HbR/HbO) changes for fNIRS. The correlation of EEG spectral features with fNIRS HbO/HbR features were then evaluated to assess the mechanisms of ALS on the electrical (EEG)-vascular (fNIRS) interrelationships. We observed overall smaller increases in EEG delta and theta power, decreases in beta power, reductions in HbO responses, and distortions both in early and later EEG event-related potentials in ALS subjects compared to healthy controls. While significant correlations between EEG features and HbO responses were observed in healthy controls, these patterns were absent in ALS patients. Distortions in both electrical and hemodynamic responses are speculated to be associated with cognitive deficits in ALS that center primarily on attentional and working memory processing. Our results highlight the important role of ALS non-motor dysfunctions in electrical and hemodynamic neural dynamics as well as their interrelationships. The insights obtained through this study can enhance our understanding of the underlying non-motor neural processes in ALS and enrich future diagnostic and prognostic techniques.


Predictive factors for rapid neutrophil and platelet engraftment after allogenic peripheral blood stem cell transplantation in patients with acute leukemia.
The aim of this study was to investigate predictive factors for rapid engraftment after allogeneic peripheral blood stem cell transplantation (alloPBSCT) in patients with acute leukemia. Two hundred sixty-two patients receiving alloPBSCT were analyzed. Subset analyses of donor stem cells were conducted using a flow cytometric method. The correlation between rapid engraftment of neutrophils, platelets, and donor stem cells doses, as well as other recipient and donor clinical factors, was analyzed. In univariate analysis, factors correlated with neutrophil engraftment (≥0.5 × 10(9)/L) by day 12 were achievement of complete remission (CR) after induction chemotherapy (CR1) before hematopoietic cell transplantation (HCT) and high numbers of CD34+ cells, CD3+ T cells, and CD3+/CD4+ T cells. Factors correlated with platelet engraftment (≥20 × 10(9)/L) by day 12 were achievement of CR1 before HCT, donor and recipient sex mismatch, and high numbers of mononuclear cells, CD34+ cells, CD3+ T cells, CD3+/CD4+ T cells, CD3+/CD8+ T cells, and CD56+ NK cells. In multivariate analysis, independent predictive factors for rapid neutrophil and platelet engraftment were CR1 before HCT (p < 0.001 and p = 0.002, respectively), high number of donor CD34+ cells (p = 0.005 and p < 0.001, respectively), and high number of CD3+ T cells (p = 0.005 and p = 0.001, respectively). In conclusion, achieving CR1 before HCT, as well as larger quantities of donor CD34+ and CD3+ T cells, may predict rapid neutrophil and platelet engraftment after PBSCT.


Prolonged chemoradiation in locally advanced carcinoma of the uterine cervix: final results of a phase II study (ESTER-1).
The aim of this phase II study was to evaluate response and toxicity of a prolonged chemoradiation regimen in patients with locally advanced cervical cancer. Three cycles of concomitant chemotherapy were used with cisplatin (20 mg/m², 2-hour intravenous infusion, days 1-4) and 5-fluorouracil (1000 mg/m², 24-hour continuous intravenous infusion, days 1-4) administered at weeks 1, 5, and 9 of radiotherapy. In combination, radiotherapy was delivered to a planning target volume (PTV) defined as the CTV (clinical target volume) plus 8 mm. The CTV was defined as follows: gross tumor volume, upper half of the vagina (if not involved) or the whole vagina (if clinically involved), uterus, obturator nodes, external iliac nodes, internal iliac nodes, and the presacral nodes (cranial to S2). The prescribed dose to the PTV was 50 Gy, 2 Gy/fraction (ICRU 62) delivered in 25 fractions with a 2-week break at 20Gy and 40 Gy (split-course technique). Early and late toxicity was assessed according to the RTOG and RTOG/EORTC toxicity scales. Perioperative toxicity was evaluated according to the Chassagne classification of surgical complications. A total of 25 patients were included in this study. Median age was 52 years (range, 28-69). Clinical stage was: IB2-II: 19 patients (76%), III-IVA: 6 patients (24%). All patients completed the prescribed dose of chemoradiation and were evaluated 4 weeks after the end of treatment. Complete and partial clinical local response was observed in 4 and 19 patients, respectively (totaling 92% of clinical responses). About 32% of patients experienced grade 3 to 4 toxicity, in particular, grade 3 or 4 hematological toxicity was observed in 7 patients and 1 patient developed grade 3 genitourinary toxicity. No patients developed grade 3 gastrointestinal toxicity or skin toxicity. Of total, 22 patients (88%) underwent radical hysterectomy. Seven patients (28%) showed a complete response (CR) to treatment, and 7 patients (28%) showed microscopic residual disease (μPR), totaling 14 patients (56%) complete/partial microscopic responses. Perioperative morbidity was higher than reported in historical controls especially in terms of tissue fibrosis (64%) and perioperative urinary toxicity (14%). Actuarial 2-year local control, disease-free survival, and overall survival were 65.5%, 61.8%, and 80.8%, respectively. A prolonged treatment with more chemotherapy courses does not improve tumor response and increases the risk of perioperative complication. This treatment regimen, considering the low incidence of acute gastrointestinal toxicity, might be tested in the adjuvant setting.


PLA-MoRe: A Protein-Ligand Binding Affinity Prediction Model via Comprehensive Molecular Representations.
Accurately predicting the binding affinity of protein-ligand pairs is an essential part of drug discovery. Since wet laboratory experiments to determine the binding affinity are expensive and time-consuming, several computational methods for binding affinity prediction have been proposed. In the representation of compounds, most methods only focus on the structural properties such as SMILES and ignore the bioactive properties. In this study, we proposed a novel model named PLA-MoRe to predict protein-ligand binding affinity, which represents compounds based on both structural and bioactive properties and mainly contains three feature extractors. First, a structure feature extractor based on the graph isomorphism network was constructed to learn the representations of the molecular graphs. Second, we designed an Autoencoder-based bioactive feature extractor to integrate the multisource bioactive information including chemical, target, network, cellular, and clinical. The above two parts aimed to learn representations of compounds in terms of structures and bioactivities, respectively. Then, we constructed a sequence feature extractor to learn embeddings for protein sequences. The output of the three extractors was concatenated and fed into a fully connected network for affinity prediction. We compared PLA-MoRe with three state-of-the-art methods, and an ablation study was conducted to test the role of each part of the model. Further attention visualization showed that our model had the potential to locate the binding sites, which might help explain the mechanism of interaction. These results prove that PLA-MoRe is competitive and reliable. The resource codes are freely available at the GitHub repository https://github.com/QingyuLiaib/PLA-MoRe.


Adipokinetic hormone (Pyrap-AKH) enhances the effect of a pyrethroid insecticide against the firebug Pyrrhocoris apterus.
Adipokinetic hormones (AKHs) are insect neuropetides controlling stress situations including those elicited by insecticide treatment. The effect of Pyrap-AKH on the mortality of the firebug Pyrrhocoris apterus (L.) treated with the insecticide permethrin (Ambush 25 EC) was studied. Coinjection of 50 ng permethrin with 80 pmol Pyrap-AKH induced a significant 2.3-fold increase in bug mortality compared with the insecticide alone. The results were confirmed by topical coapplication of both agents (400 ng and 80 pmol respectively). Injections of 50 and 100 ng permethrin elicited a significant increase in the AKH level in CNS and the haemolymph. The results indicate an involvement of AKH in stress response to permethrin. The enhanced effect of insecticide by AKH treatments probably results from the stimulatory role in bug metabolism: carbon dioxide production was increased 3.5- and 2.5-fold respectively 1 and 3 h after permethrin treatment, and 4.3- and 3.4-fold after the permethrin plus AKH cotreatment, compared with the control. The elevation of metabolism could intensify the permethrin action by its faster penetration into tissues and by stimulation of biochemically active cells, and could be a reason for enhanced action of permethrin after its cotreatment with Pyrap-AKH.


Effect of potassium hydrogen difluoride in zirconia-to-resin bonding.
The objective of this study was to compare potassium hydrogen difluoride (KHF2) etching for zirconia with commonly used surface roughening and chemical bonding methods (silane, MDP-monomer primer) for resin-based luting cement bonding to zirconia. Zirconia specimens were divided into six groups (n=10) according to surface treatment and bonding procedures, with and without thermocycling (6,000 cycles, 5-55ºC): 1) air-borne particle abrasion with alumina+MDP-monomer (ABP), 2) air-borne particle abrasion with silica-coated trialuminium trioxide+silane (ABPR-S) and 3) KHF2 etching+silane (ETC). Surface roughness and bond strength (SBS-test) for dry and thermocycled specimens were measured. SBS did not vary statistically between the dry groups, but thermocycling decreased the bond strengths of all the tested methods (p<0.05). After thermocycling, ABP had statistically significantly lower bond strength values compared to ABPR-S and ETC (p<0.05). Etching method with KHF2 did not provide better bonding capacity to previously introduced and commonly adopted bonding methods.


Association of atherosclerotic cerebral infarction and human leukocyte antigen-DRB in a North Chinese Han population.
Evidence from experimental and genetic studies suggest the existence of a potential link between the polymorphisms of human leukocyte antigen class II gene (HLA-DR) and ischemic stroke. This study addressed the association of HLA-DR gene with atherosclerotic cerebral infarction (ACI) in a North Chinese Han population. The genotyping of HLA-DRB1 was determined by standard techniques based on polymerase chain reaction and sequence-specific oligonucleotides hybridization in a gene chip. The relative risk (RR) of HLA-DRB1 FNx01 04 and HLA-DRB1 FNx01 03 (17) in patients with ACI and their first-degree relatives were significantly higher than those in the control group (RR=2.56 and 18.77, respectively; P <0.05). In contrast, the RR of HLA-DRB1 FNx01 12 was dramatically reduced in patients with ACI in relation to healthy controls (RR=0.17; P <0.01). These data indicate that the polymorphisms in HLA-DRB1 may influence the risk of ACI in the North Han population of China. Further studies are necessary to validate the observation in larger samples.


Functional Analysis and Genome Mining Reveal High Potential of Biocontrol and Plant Growth Promotion in Nodule-Inhabiting Bacteria Within Paenibacillus polymyxa Complex.
Bacteria belonging to the genus Paenibacillus were frequently isolated from legume nodules. The nodule-inhabiting Paenibacillus as a resource of biocontrol and plant growth-promoting endophytes has rarely been explored. This study explored the nodule-inhabiting Paenibacillus' antifungal activities and biocontrol potentials against broad-spectrum important phytopathogenic fungi. We collected strains which were isolated from nodules of Robinia pseudoacacia, Dendrolobium triangulare, Ormosia semicastrata, Cicer arietinum, Acacia crassicarpa, or Acacia implexa and belong to P. peoriae, P. kribbensis, P. endophyticus, P. enshidis, P. puldeungensis, P. taichungensis, or closely related to P. kribbensis, or P. anseongense. These nodule-inhabiting Paenibacillus showed diverse antagonistic activities against five phytopathogenic fungi (Fusarium graminearum, Magnaporthe oryzae, Rhizoctonia solani, Sclerotinia sclerotiorum, and Botrytis cinerea). Six strains within the P. polymyxa complex showed broad-spectrum and potent activities against all the five pathogens, and produced multiple hydrolytic enzymes, siderophores, and lipopeptide fusaricidins. Fusaricidins are likely the key antimicrobials responsible for the broad-spectrum antifungal activities. The nodule-inhabiting strains within the P. polymyxa complex were able to epiphytically and endophytically colonize the non-host wheat plants, produce indole acetic acids (IAA), and dissolve calcium phosphate and calcium phytate. P. peoriae strains RP20, RP51, and RP62 could fix N2. P. peoriae RP51 and Paenibacillus sp. RP31, which showed potent plant colonization and plant growth-promotion competence, effectively control fungal infection in planta. Genome mining revealed that all strains (n = 76) within the P. polymyxa complex contain ipdC gene encoding indole-3-pyruvate decarboxylase for biosynthesis of IAA, 96% (n = 73) contain the fus cluster for biosynthesis of fusaricidins, and 43% (n = 33) contain the nif cluster for nitrogen fixation. Together, our study highlights that endophytic strains within the P. polymyxa complex have a high probability to be effective biocontrol agents and biofertilizers and we propose an effective approach to screen strains within the P. polymyxa complex.


Soluble vascular cell adhesion molecule-1 and magnesium sulfate with nifedipine treatment in Indonesian women with severe pre-eclampsia.
Endothelial cell activation in pre-eclampsia is associated with elevated soluble vascular cell adhesion molecule-1 (sVCAM-1) levels. The objective of the study was to determine whether sVCAM-1 levels in Indonesian women with pre-eclampsia were similar to other ethnic studies and to determine the effects of magnesium sulfate with nifedipine on blood pressure. A total of 61 pregnant women were admitted, who had normal pregnancy (n = 25) and severe pre-eclampsia (n = 36). Blood sampling was performed at admission to the study, 1 h after placental separation, and 24 h postpartum. sVCAM-1 and blood pressure levels were determined. The mean ages in normal pregnancy (n = 25) and in severe pre-eclampsia (n = 36) are 30.0 ± 3.4 years and 27.1 ± 6.1 years, respectively. Significantly elevated sVCAM-1 was seen in pre-eclampsia. No significant variation in sVCAM-1 levels during the study periods was seen in both groups of cohorts. Magnesium sulfate infusion and nifedipine significantly lowered the blood pressure level. Elevated sVCAM-1 levels were also seen in Indonesian women with severe pre-eclampsia. The placenta may not be the only source of elevated sVCAM-1 and that endothelial dysfunction persists beyond the postpartum period. Magnesium sulfate together with nifedipine significantly lowered blood pressure. The determination of elevated sVCAM-1 in pregnancy as a risk marker for endothelial dysfunction is therefore suggested.


Serum immunoglobulin G immune response to Helicobacter pylori antigens in Mongolian gerbils.
The Mongolian gerbil model for Helicobacter pylori infection is an animal model that mimics human disease. We examined the serum immune response to H. pylori infection in gerbils by enzyme-linked immunosorbent assay (ELISA) and Western blotting, both with whole-cell (H. pylori) extracts. A total of 66 7-week-old specific-pathogen-free male gerbils were inoculated orogastrically with H. pylori strain ATCC 43504. Sera were collected 1, 2, 4, 8, 12, 26, 38, and 52 weeks after H. pylori inoculation. Sixty-nine noninfected gerbils and their sera were used as controls. The specificity of the ELISA was 95.7%. The frequency of seropositivity increased over time: 2 of 10 (20%), 7 of 10 (70%), and 7 of 7 (100%) samples of sera from inoculated gerbils were positive for H. pylori at 2, 4, and 8 weeks postinoculation, respectively. Western blot assays showed that the primary immunoglobulin G (IgG) response against low-molecular-mass (25-, 30-, and 20-kDa) proteins appeared after a lag period of 2 to 8 weeks after inoculation. Antibodies against 160-, 150-, 110-, 120-, 80-, 66-, and 63-kDa proteins were observed 12 weeks after inoculation. The early reactive 30-kDa protein was identified as a urease alpha subunit by N-terminal amino acid sequencing. After 26 weeks, two groups of animals could be distinguished: one group developed ulcers (n = 5), and the other developed hyperplastic polyps without ulcers (n = 19). Gerbils in the gastric ulcer group showed significantly higher serum anti-H. pylori IgG levels than did gerbils in the hyperplastic group (P = 0.001) as measured by ELISA. Furthermore, a higher proportion of animals developed antibodies to H. pylori proteins of 26, 25, and 20 kDa in the ulcer group than those animals with hyperplastic polyps (75 to 100% versus 17 to 50%) in Western blot assays. These results highlight the importance of the immune response of the host in the development of H. pylori-related gastric lesions.


Medical knowledge evolution query constraining aspects.
In this paper we present a first analysis towards better understanding of the query constraining aspects of knowledge, as expressed in the most used public medical bibliographic database MEDLINE. Our results indicate, possibly not surprising, that new terms occur, but also that traditional terms are replaced by more specific ones or even go out of use as they become common knowledge. Hence, as knowledge evolve over time, search methods may benefit from becoming more sensitive to knowledge expression, to enable finding new, as well as older, relevant database contents.


Carbon dioxide assimilation by leaves, isolated chloroplasts, and ribulose bisphosphate carboxylase from spinach.
The relationship between rate of photosynthesis and CO(2) concentration has been reinvestigated using isolated spinach (Spinacia oleracea) chloroplasts. The apparently low CO(2) concentration required for half-maximal photosynthesis is shown to result partly from a ceiling imposed by electron transport. In double reciprocal plots of rate against CO(2) concentration, this ceiling results in departures from linearity at high CO(2) concentrations. If these rate limitations are disregarded in extrapolation the "true" CO(2) concentration required for half maximal carboxylation by intact chloroplasts is approximately 46 mum (CO(2)).When assayed under comparable conditions, ribulose bisphosphate carboxylase from these chloroplasts also shows an apparent Km (CO(2)) of approximately 46 mum, suggesting that its characteristics are not modified by extraction. An improved assay for ribulose bisphosphate carboxylase yielded rates of carboxylation considerably higher than those previously reported, the highest maximal velocities recorded approaching 1000 mumoles CO(2) fixed mg(-1) chlorophyll hr(-1) at 20 C. With such Km and V(max), values the carboxylase would be able to achieve, at concentrations of CO(2) less than atmospheric, rates of CO(2) fixation equal to those displayed by the parent tissue or by the average plant under favorable conditions in its natural environment.


Pre-harvest methyl jasmonate treatments increase antioxidant systems in lemon fruit without affecting yield or other fruit quality parameters.
Jasmonic acid (JA) and its volatile derivative methyl jasmonate (MeJA) are hormones involved in the regulation of many processes in plants and act (when applied as a post- or pre-harvest treatment) to increase fruit bioactive compounds with antioxidant potential. However, there is no literature available regarding the effect of pre-harvest MeJA treatment on lemon fruit antioxidant systems, which was the aim of the present study. MeJA treatment (0.1, 0.5 and 1.0 mmol L-1 ) increased antioxidant compounds, such as phenolics, in the juice and flavedo of 'Fino' and 'Verna' lemons at harvest, with the most effective concentration being 0.1 mmol L-1 in both cultivars. In addition, catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX) activities were also increased by MeJA treatment, with the highest increases being also found with 0.1 mmol L-1 . The increases in APX and CAT were maintained from one treatment to another during fruit development on the tree, whereas the increase on POD disappeared after 8-10 days of each treatment. For both antioxidant systems, the highest increases were found in lemon harvested at the commercial ripening stage. By contrast, crop yield, fruit ripening process and quality parameters were generally not affected by MeJA treatment. Preharvest MeJA treatment could be a useful tool for increasing antioxidant potential and the health beneficial effects of lemon fruit consumption, given the relationship between these properties and phenolic content. Moreover, the increased concentration of phenolics and the activity of antioxidant enzymes in the flavedo of MeJA treated fruit could increase lemon tolerance to chilling injury and decay during postharvest storage. © 2019 Society of Chemical Industry.


Microcephaly: general considerations and aids to nosology.
Microcephaly is defined as an occipito-frontal head circumference (OFC) 2 or more standard deviations below the mean for age and sex using the new Roche et al. [Pediatrics 1987;79:706-712] charts, and corrected for parental OFC by the method of Weaver and Christian [J Pediatr 1980;96:990-994]. "Relative" microcephaly, i.e., a small head on a small child, may be associated with a much better intellectual prognosis than absolute microcephaly, although the average IQ of children with absolute microcephaly ascertained in a normal school system is normal when compared with that of appropriate control children. "Primary" microcephaly means an abnormal OFC at birth (corrected for gestational age and length), and "secondary" microcephaly a normal birth OFC with later, acquired microcephaly due to deceleration of brain growth reflecting infection, trauma, intoxication, metabolic disease, the Rett syndrome, or a true CNS degenerative disease. Some cases of syndromal microcephaly may be associated with normal intelligence including some "primordial dwarfs," children with Dubowitz syndrome, FAS, mild SC-Roberts syndrome, and an occasional Brachmann-de Lange individual. The nosology of (syndromal) microcephaly is extraordinarily complex and requires the assistance of special library resources and information retrieval expertise. At a minimum, it requires McKusick's Catalog of Mendelian Inheritance in Man (MIM); however, we find that our work is greatly enhanced by recently developed electronic databases such as MIM-online (OMIM), POSSUM, SYNDROME, and MEDLINE, as well. Three groups of syndromal and non-syndromal microcephaly are discussed selectively in order to illustrate the marvels of pleiotropy in human development and its abnormalities and the difficulties encountered in splitting and lumping entities with overlapping manifestations.


Height and body fatness and colorectal cancer risk: an update of the WCRF-AICR systematic review of published prospective studies.
There is no published dose-response meta-analysis on the association between height and colorectal cancer risk (CRC) by sex and anatomical sub-site. We conducted a meta-analysis of prospective studies on the association between height and CRC risk with subgroup analysis and updated evidence on the association between body fatness and CRC risk. PubMed and several other databases were searched up to November 2016. A random effects model was used to calculate dose-response summary relative risks (RR's). 47 studies were included in the meta-analyses including 50,936 cases among 7,393,510 participants. The findings support the existing evidence regarding a positive association of height, general and abdominal body fatness and CRC risk. The summary RR were 1.04 [95% (CI)1.02-1.05, I² = 91%] per 5 cm increase in height, 1.02 [95% (CI)1.01-1.02, I² = 0%] per 5 kg increase in weight, 1.06 [95% (CI)1.04-1.07, I² = 83%] per 5 kg/m2 increase in BMI, 1.02 [95% (CI)1.02-1.03, I² = 4%] per 10 cm increase in waist circumference, 1.03 [95% (CI)1.01-1.05, I² = 16%] per 0.1 unit increase in waist to hip ratio. The significant association for height and CRC risk was similar in men and women. The significant association for BMI and CRC risk was stronger in men than in women. The positive association between height and risk of CRC suggests that life factors during childhood and early adulthood might play a role in CRC aetiology. Higher general and abdominal body fatness during adulthood are risk factors of CRC and these associations are stronger in men than in women.


Measurement of velopharyngeal movements induced by isolated stimulation of levator veli palatini and pharyngeal constrictor muscles.
The levator veli palatini (LVP) and superior pharyngeal constrictor muscles (PC) close the velopharynx. However, for the velopharyngeal movements to be understood in detail, each muscle contraction must be analyzed precisely. This study was performed to clarify the velopharyngeal movement which was induced by a single muscle contraction, LVP or PC. Using a nasopharyngeal fiberscope, we analyzed the velopharyngeal movement associated with the contraction of the LVP and PC muscles in mongrel dogs. To elicit the maximal contraction of each muscle, we applied repetitive electrical stimulation to each peripheral nerve efferent to the LVP or PC muscle. Stimulation with a frequency of 77 Hz and 83 Hz induced maximal tension in the LVP and PC muscles, respectively, in isometric contraction. In a second series of experiments, fiberscopic views of the velopharyngeal movements associated with each muscle's maximal contraction were recorded. The degree of closure was calculated at several sections. The LVP muscle pulled the caudal fourth of the soft palate, while the PC projected the posterior wall at the level of the caudal end of the soft palate. The PC muscle also projected the lateral wall of the velopharynx. The effect of LVP contraction on the lateral wall was very small. These results show that the velopharyngeal movement associated with LVP contraction is very different from that with PC contraction.


Stress Hyperglycemia Is Associated With an Increased Risk of Subsequent Development of Diabetes Among Bacteremic and Nonbacteremic Patients.
Stress hyperglycemia is associated with an increased risk of diabetes among survivors of critical illness. We investigated whether patients without diabetes hospitalized for bacteremia or nonbacteremic diseases with transient stress hyperglycemia would have a higher risk of subsequent diabetes development compared with those who remained normoglycemic. This retrospective observational study was conducted on 224,534 in-patients with blood culture records. Stress hyperglycemia was defined based on the highest random glucose level ≥7.8 mmol/L during the index admission period. Diagnosis of diabetes, as the primary end point of interest, was defined based on diagnostic codes, blood test results, or medication records. Differences in cumulative incidence and hazard ratios (HRs) of diabetes between groups were assessed using the Kaplan-Meier estimator and Cox regression. After exclusion of patients with preexisting or undiagnosed diabetes or indeterminate diabetes status and propensity score matching, bacteremic patients with stress hyperglycemia had a significantly higher cumulative incidence of diabetes (HR 1.7, 95% CI 1.2-2.4) compared with those who remained normoglycemic. Stress hyperglycemia was further confirmed to be a diabetes predictor independent of age, sex, comorbidity, and other serological markers. For the nonbacteremic patients, stress hyperglycemia was similarly associated with a higher cumulative incidence of diabetes (HR 1.4, 95% CI 1.2-1.7). Hospitalized patients with transient stress hyperglycemia had a higher risk of subsequent diabetes development compared with their normoglycemic counterparts. Recognition of an increased risk of diabetes in these patients can allow early detection and monitoring in their subsequent follow-ups.


Decreased phosphorus metabolite concentrations and alkalosis in chronic cerebral infarction.
A study was performed to determine quantitatively the alterations in phosphorus metabolite concentrations and pH in regions of the human brain damaged by chronic stroke. Image-guided phosphorus-31 magnetic resonance spectroscopy was performed on the brains of eight healthy subjects and six patients with cerebral infarction of more than 3 months duration. Phosphorus metabolite concentrations in infarcted regions were reduced 8%-67%. Significant decreases occurred in phosphomonoester (PME), phosphodiester (PDE), and adenosine triphosphate (ATP) concentrations, while inorganic phosphate (Pi) and phosphocreatine (PCr) concentrations showed smaller, nonsignificant decreases. The PCr/ATP ratio was significantly increased, while the ATP/Pi ratio was somewhat lower. The phospholipid ratio PDE/PME was also significantly increased, while the ratios of phospholipid (PME, PDE) to phosphate (PCR, Pi) metabolites were significantly decreased. The pH of the infarcted region indicated significantly more alkalinity than in the normal brain. The results suggest that chronic stroke is associated with significant changes in brain metabolite concentrations and pH that are different from those reported for other brain diseases.


Efficiency of snake sampling methods in the Brazilian semiarid region.
The choice of sampling methods is a crucial step in every field survey in herpetology. In countries where time and financial support are limited, the choice of the methods is critical. The methods used to sample snakes often lack objective criteria, and the traditional methods have apparently been more important when making the choice. Consequently researches using not-standardized methods are frequently found in the literature. We have compared four commonly used methods for sampling snake assemblages in a semiarid area in Brazil. We compared the efficacy of each method based on the cost-benefit regarding the number of individuals and species captured, time, and financial investment. We found that pitfall traps were the less effective method in all aspects that were evaluated and it was not complementary to the other methods in terms of abundance of species and assemblage structure. We conclude that methods can only be considered complementary if they are standardized to the objectives of the study. The use of pitfall traps in short-term surveys of the snake fauna in areas with shrubby vegetation and stony soil is not recommended.


Impaired bone marrow-derived macrophage differentiation in vitamin D deficiency.
The calcium-regulating hormone, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), is also recognized as an immunomodulator. In vitro addition of the hormone to bone marrow-derived macrophages (BMDMs) results in a decreased proliferation and an increased differentiation. In the present work we compare the in vitro differentiation of BMDMs derived from vitamin D-depleted and -repleted mice (-D and +D BMDMs, respectively). -D BMDMs proliferate in vitro slower than +D BMDMs. Addition of the hormone to BMDM cultures inhibited the rate of their proliferation, which was more pronounced in low-density cultures. The ability of mononuclear phagocytes to produce reactive oxygen metabolites is an important element in the microbicidal functions of these cells. We found that -D BMDMs produce less H2O2 than +D BMDMs, which was corrected by the in vitro addition of 1,25(OH)2D3. Analyses of various macrophage-specific surface antigens revealed a reduction in their expression on -D BMDMs. In vitro addition of 1,25(OH)2D3 to BMDM cultures increased the expression of these antigens. The activity of the lysosomal enzyme acid phosphatase was similarly affected by vitamin D deficiency and by the in vitro addition of the hormone. Thus, vitamin D deficiency is associated with impaired maturation of BMDMs suggesting that the hormone is a natural modulator of macrophage maturation.


Click chemistry with polymers, dendrimers, and hydrogels for drug delivery.
During the last decades, great efforts have been devoted to design polymers for reducing the toxicity, increasing the absorption, and improving the release profile of drugs. Advantage has been also taken from the inherent multivalency of polymers and dendrimers for the incorporation of diverse functional molecules of interest in targeting and diagnosis. In addition, polymeric hydrogels with the ability to encapsulate drugs and cells have been developed for drug delivery and tissue engineering applications. In the long road to this successful story, pharmaceutical sciences have been accompanied by parallel advances in synthetic methodologies allowing the preparation of precise polymeric materials with enhanced properties. In this context, the introduction of the click concept by Sharpless and coworkers in 2001 focusing the attention on modularity and orthogonality has greatly benefited polymer synthesis, an area where reaction efficiency and product purity are significantly challenged. The purpose of this Expert Review is to discuss the impact of click chemistry in the preparation and functionalization of polymers, dendrimers, and hydrogels of interest in drug delivery.


Selective enrichment of azide-containing peptides from complex mixtures.
A general method is described to sequester peptides containing azides from complex peptide mixtures, aimed at facilitating mass spectrometric analysis to study different aspects of proteome dynamics. The enrichment method is based on covalent capture of azide-containing peptides by the azide-reactive cyclooctyne (ARCO) resin and is demonstrated for two different applications. Enrichment of peptides derived from cytochrome c treated with the azide-containing cross-linker bis(succinimidyl)-3-azidomethyl glutarate (BAMG) shows several cross-link containing peptides. Sequestration of peptides derived from an Escherichia coli proteome, pulse labeled with the bio-orthogonal amino acid azidohomoalanine as substitute for methionine, allows identification of numerous newly synthesized proteins. Furthermore, the method is found to be very specific, as after enrichment over 87% of all peptides contain (modified) azidohomoalanine.


[Localization of impacted maxillary canines by palpation and orthopantomography].
In orthodontic practice occurred problems associated with labio-palatal localization of impacted permanent maxillary canines. Not always is possible to determinate labio-palatal location of these teeth on the basis of palpation, therefore clinical examination aided by radiographic investigation. Orthopantomogram is the most widely used radiograph in orthodontics. The purpose of present study-test diagnostic value of palpation and orthopantomography in cases of maxillary canines impaction on the basis of evaluation of migration and inclination of impacted teeth. Thirty nine patients of Clinic of Orthodontics, Kaunas University of Medicine, orthopantomograms were available for analysis. Forty two impacted canines were investigated. Only 64% of canines could be palpated. It depended on the vertical position of the tooth. Distribution of the values of canine inclination to the middle of upper jaw was too big for more precise statistical analysis. Canines, which were medial to the 3th horizontal sector (these canines overlapped longitudinal axis of the lateral incisor), were located palatally.


Culture of troubled work groups.
Managing the troubled or conflicted work group can occupy a great deal of a manager's time and energy. One hundred fifteen nurse managers and administrators responded to a descriptive survey exploring the characteristics of the culture of troubled or conflicted work groups. Those managers and administrators also described numerous successful strategies that they had used in managing these work groups.


Bronchiolitis Obliterans Syndrome and Death in Iranian Lung Transplant Recipients: A Bayesian Competing Risks Analysis.
Bronchiolitis obliterans syndrome (BOS) is delayed allograft deterioration after lung transplant (LTX) that is clinically characterized by ≥ 20% decline from the baseline value of forced expiratory volume during the first second (FEV1). BOS is still a major obstacle limiting long-term survival post-LTX. The main aim of this study was to determine the predictors of BOS and death in Iranian LTX recipients. This retrospective cohort study included 44 LTX recipients who survived ≥ 3 months post-LTX at the Masih Daneshvari Hospital, Tehran, Iran from 2000 to 2014. The outcome was time from lung transplantation to BOS and/or death (due to all causes except BOS). We used competing risks analysis to assess the effect of other factors on the cumulative incidence function of BOS and death. We applied a Fine and Gray model with Bayesian approach. The recipients' age (Mean ± SD) was 36.7 ± 14.5 yr. 11 (25%) recipients developed BOS as the first event within the first five years post-LTX and 13 (30%) died due to all causes except for BOS. Our results showed that CMV infection was associated with a significant increase in risk of developing BOS [hazard ratio (HR) 1.22 (95% credible set: (1.01, 3.2)] controlling for other variables. Bilateral transplantation [HR (95% credible set): 2.4(1.51, 4.05)] and CMV infection [HR (95% credible set): 2.02 (1.67, 2.55)] were predictors of the mortality risk. CMV infection was a predictor of BOS risk in the studied patients. Moreover, bilateral transplantation and CMV infection were significant predictors of mortality in the present sample. Multi-center studies with larger sample sizes are required to better study the other risk factors, and the pathophysiologic mechanisms of BOS.


The Arp2/3 complex: a multifunctional actin organizer.
The actin-related proteins (Arps) constitute a recently characterized family of proteins, many of which function as members of multiprotein complexes. The discovery that two family members, Arp2 and Arp3, act as multifunctional organizers of actin filaments in all eukaryotes has generated much excitement. Over the past two years, newly discovered properties of the Arp2/3 complex have suggested a central role in the control of actin polymerization. First, it promotes actin assembly on the surface of the motile intracellular pathogen Listeria monocytogenes. Second, it can nucleate and cross-link actin filaments in vitro. Third, it localizes with dynamic actin-rich spots of mammalian cells suggesting a role in protrusion; it is found in cortical actin patches in the budding and fission yeasts where it may control patch movement and cortical actin function. Clearly, the complex has a central role in actin cytoskeletal function and will be the subject of much research in the coming years.


Marriage as bondage: the anomaly of the citizen wife.
The meaning of marriage for women was a hotly disputed issue in Australia during the first half of the twentieth century and beyond. Conscious of their status as citizens, interwar feminists became preoccupied with the anomalous condition of wives in a modern democracy. The provision of personal service in exchange for keep, the condition of wives was defined as an anachronism, a political relic, a form of feudal bondage. Feminists campaigned for the economic independence of married women, demanding legislation that would give wives legal ownership of household savings as well as a legal entitlement to a share in their husbands' wages.


Intensive chemotherapy for poor prognosis myelodysplasia (MDS) and secondary acute myeloid leukemia (sAML) following MDS of more than 6 months duration. A pilot study by the Leukemia Cooperative Group of the European Organisation for Research and Treatment in Cancer (EORTC-LCG).
We conducted a prospective, multicenter pilot study of remission induction therapy in patients with poor prognosis MDS and AML evolving from a preceding phase of MDS. Fifty evaluable patients from 15 institutions were treated with one or two remission-induction courses consisting of i.v. idarubicin 12 mg/m2/day on days 1, 2, and 3 combined with a continuous i.v. infusion of cytarabine of 200 mg/m2/day on days 1 to 7. Of the 27 complete remitters (54%), 23 received a consolidation course which was identical to the remission-induction course except for the idarubicin 12 mg/m2 which was given on day 1 only. Fifteen patients received maintenance therapy consisting of six courses of cytarabine 10 mg/m2, s.c. twice daily, for 14 days. Two complete remitters were allografted and five patients received an ABMT. The median survival of all 50 treated patients was 14 months. The median duration of disease-free survival was 11 months with two patients in CR more than 2 years after entering CR. Twenty-four of the 27 remitters have relapsed. Four patients died during remission-induction therapy, but no patient died as a result of persisting hypoplasia. No fatal complications occurred during the consolidation and maintenance courses. Age and stage of disease had no significant impact on CR rate nor on remission duration. The CR rate was significantly (P = 0.03) higher in patients with only normal metaphases compared to patients with cytogenetic abnormalities. The DFS at 2 years was 33 vs 8%, respectively, for patients without or with cytogenetic abnormalities (P = 0.02). This study shows that patients below the age of 60 years with poor risk features are candidates for treatment with combination chemotherapy. A complete remission rate of more than 50% may be expected. Maintaining remission after remission-induction chemotherapy is a difficult issue. Patients not eligible for allogeneic BMT may be treated with intensive post-remission chemotherapy or autologous BMT.


Outcome analysis of patients with vasculitis associated with antineutrophil cytoplasmic antibodies.
Objective scoring systems of disease activity and disease-associated damage have proven useful in the management of patients with systemic vasculitis. We used the recently designed Birmingham vasculitis activity score (BVAS; maximum score 63) and vasculitis damage index (VDI; maximum score 59) to assess initial activity and long-term damage, respectively, in ANCA positive patients from one center over a 3-year period. Thirty-two patients with ANCA vasculitis were identified and analyzed as an historic cohort. The median BVAS for all vasculitis patients at first presentation was 19 (range 6 - 36). Patients with Wegener's granulomatosis had a significantly higher total score and respiratory BVAS score compared to the 15 with microscopic polyangiitis. The majority of patients received standard cyclophosphamide/steroid treatment. At the end of follow-up (mean 24.9 months), 4 patients had died; all patients had evidence of permanent organ damage. The median total VDI score at last follow-up was 4.0 (range 0-11), with no differences between patients with Wegener's granulomatosis and microscopic polyangiitis. The VDI was not associated with the number of relapses. A high initial BVAS was found to correlate with a later high vasculitis damage index (r = 0.56). Initial renal or respiratory involvement was also associated with longterm damage in the same organ system. Although mortality from ANCA-associated vasculitis has decreased, morbidity remains a common problem. High early-disease activity may identify patients at high risk of long-term organ damage, allowing more effective individualized therapy. This hypothesis requires validation in a prospective, controlled study.


The effect of limitation in ankle dorsiflexion on knee joint function. A pilot study.
A restriction in ankle dorsiflexion is a common complication of ankle fractures. This kind of dysfunction, if severe, can significantly influence gait. A restriction in ankle dorsiflexion (forward movement of the shin relative to the foot) can cause, among others, hyperextension of the knee during the stance phase. The length of leading leg step is shortened and alternant walk downstairs is very difficult. The aim of this study was to examine the correlation between the range of dorsiflexion in the ankle joint and the range of extension (hyperextension) in the knee joint. The study enrolled 17 patients after ankle joint fractures treated conservatively or surgically. The extension ranges of motion in the ankle and knee joints were assessed by goniometry to compare these values in injured vs. healthy limbs. Non-parametric methods (the Wilcoxon signed-rank test) were used for the analysis. The results showed limitation in ankle dorsiflexion in the fractured limb, which amounted to 4.40 vs. 16.00 in healthy limbs in all patients. This difference was statistically significant (p<0.001). There was also a significantly (p<0.001) greater range of knee hyperextension on the side of injury (5.00 vs. 1.90 in healthy limbs). 1. Post-traumatic restriction of ankle dorsiflexion can cause knee joint overload. 2. Examinations of knee function during walking should be carried out in patients with trauma-related dysfunctions of the ankle joint in order to prevent secondary musculoskeletal abnormalities.


Rapid Communication: Evaluation of methane inhibitor 3-nitrooxypropanol and monensin in a high-grain diet using the rumen simulation technique (Rusitec).
The objective of this study was to evaluate the effects of 3-nitrooxypropanol (NOP), a known methane (CH) inhibitor; the ionophore monensin (MON); and their combination on in vitro CH production in a high-grain diet (85% barley grain, 10% barley silage, and 5% vitamin-mineral supplement; DM basis) using a rumen simulation technique (Rusitec). Sixteen fermentation vessels in 2 Rusitec apparatuses (blocks) were used in a completely randomized block design with 4 treatments: Control, NOP (200 µg/g DM), MON (200 µg/g DM), and the combination of 200 µg NOP/g DM and 200 µg MON/g DM (NOP + MON). Two fermenters within each apparatus were randomly assigned to a treatment. Treatments were mixed with 10 g of substrate and supplied on a daily basis. The study included an 8-d adaptation period without treatment supplementation and a 6-d period for addition of treatments. Dry matter disappearance, pH, and total VFA were not affected by treatment ( ≥ 0.34). Acetate proportion was decreased by 8.3% and 14.9% with NOP and NOP + MON ( < 0.01), respectively; however, propionate proportion was not affected by treatment ( = 0.44). The acetate to propionate ratio was lowered by 21.1% with the combination of NOP and MON ( = 0.02), whereas ammonia-N concentration was not affected by treatment ( = 0.50). Total gas production was unaffected ( = 0.50), but CH production decreased by 77.7% and 75.95% ( < 0.01) with NOP and NOP + MON addition, respectively. Concurrently, H gas production increased by 131.3% and 185.6% ( = 0.01) with NOP and NOP + MON treatments, respectively. The copy number of methanogens was decreased in both solid and liquid phases ( < 0.01) with NOP and NOP + MON treatments. Despite the combination of NOP + MON showing the greatest decrease in acetate molar proportion and acetate to propionate ratio, it did not further inhibit CH beyond the effect of NOP alone. The decrease in CH emissions with treatments that included NOP occurred along with a decrease in the copy number of methanogens associated with the solid and liquid phases, confirming the inhibitory effects of NOP on these microorganisms. In conclusion, the combined effects of NOP and MON on CH mitigation did not exceed the effect of NOP alone when using a high-grain diet in vitro.


Evidence-Based Practices in Facial Reanimation Surgery.
After studying this article, the participant should be able to: 1. Describe the causes and preoperative evaluation of facial paralysis. 2. Discuss techniques to restore corneal sensation and eyelid closure, elevation of the upper lip for smile, and depression of the lower lip for lip symmetry. 3. Outline treatment goals, surgical treatment options, timing of repair, and other patient-specific considerations in appropriate technique selection. Congenital facial paralysis affects 2.7 per 100,000 children; Bell palsy affects 23 per 100,000 people annually; and even more people are affected when considering all other causes. Conditions that impair facial mimetics impact patients' social functioning and emotional well-being. Dynamic and static reconstructive methods may be used individually or in concert to achieve adequate blink restoration, smile strength and spontaneity, and lower lip depression. Timing of injury and repair, patient characteristics such as age, and cause of facial paralysis are all considered in selecting the most appropriate reconstructive approach. This article describes evidence-based management of facial paralysis.


Biochemical characterization, molecular cloning and expression of laccases - a divergent gene family - in poplar.
The nature of the enzyme(s) involved in the dehydrogenative polymerization of lignin monomers is still a matter of debate. Potential candidates include laccases which have recently received attention due to their capacity to oxidize lignin monomers and close spatial and temporal correlation with lignin deposition. We have characterized two H2O2-independent phenoloxidases with approximate molecular masses of 90 kDa and 110 kDa from cell walls of Populus euramericana xylem that are capable of oxidizing coniferyl alcohol. The 90-kDa protein was purified to apparent homogeneity and extensively characterized at the biochemical and structural levels. To our knowledge, this is the first report of a plant laccase purified to homogeneity from a lignifying tissue of an angiosperm. The cDNA clones corresponding to the 90-kDa and 110-kDa proteins, lac90 and lac110, were obtained by a PCR-based approach using specific oligonucleotides derived from peptide sequences. Sequence analysis indicated that lac90 and lac110 encoded two distinct laccases. In addition, heterologous screening using an Acer pseudoplatanus laccase cDNA enabled us to obtain three additional cDNAs (lac1, lac2, lac3) that did not correspond to lac90 and lac110. The five laccase cDNAs correspond to a highly divergent multigene family but Northern analysis with gene-specific probes indicated that all of the genes are exclusively and abundantly expressed in stems. These results highlight the polymorphism of plant laccases by an integrated biochemical and molecular approach, and provide the tools that will enable us to clearly determine the function of these enzymes in plants by molecular and genetic approaches.


Effect of rosiglitazone versus insulin on the pancreatic beta-cell function of subjects with type 2 diabetes.
In a previous study, we found observational evidence of improvement in beta-cell function when rosiglitazone was added to a failing oral antihyperglycemic regimen consisting of sulfonylureas and metformin. To confirm our previous observations, we designed and performed a prospective, randomized, and controlled study. A total of 17 subjects with type 2 diabetes, inadequately controlled on a maximized oral antihyperglycemic double regimen of glimepiride and metformin, were randomized to the addition of rosiglitazone or insulin to their treatment regimens for a period of 6 months. At baseline and at 6 months, the following were performed: measurement of fasting plasma glucose, fasting proinsulin, and insulin levels; frequently sampled intravenous glucose tolerance test; and glucagon stimulation test for C-peptide. Nine subjects were randomized to the addition of 8 mg rosiglitazone, and eight subjects were randomized to the addition of one injection of insulin (premixed 70/30) before their evening meal. The treatment groups were well matched for age, duration of diabetes, and BMI. Most important, the HbA(1c) was well matched between groups before treatment (8.7 +/- 0.3 and 9.0 +/- 0.3%; NS) and at the end of the 6 months (7.8 +/- 0.5 and 7.8 +/- 0.3%; NS). After 6 months, at the end of the study, there was a significant improvement in acute insulin response to glucose in the rosiglitazone group (+15.3 microIU x ml(-1) x 10 min(-1); P < 0.001) that led to an increase in the disposition index from 0.18 at baseline to 4.18 at 6 months (P = 0.02). Furthermore, at the end of the study, the proinsulin-to-insulin ratio had decreased in the rosiglitazone group by 36% (P = 0.03) but did not change significantly in the insulin treatment group. Rosiglitazone, but not insulin, induced a recovery of pancreatic beta-cell function, as evidenced by the restoration of the first-phase insulin response to glucose, improvement in the disposition index, and a decrease in the proinsulin-to-insulin ratio in subjects with type 2 diabetes in whom oral antihyperglycemic therapy failed. This improvement was independent of the correction of glucotoxicity.


Comprehensive quality evaluation of Fructus Schisandrae using electrospray ionization ion trap multiple-stage tandem mass spectrometry coupled with chemical pattern recognition techniques.
Electrospray ionization ion trap multiple-stage tandem mass spectrometry (ESI-MS(n)) was used to evaluate Fructus Schisandrae of similar species (Schisandra chinensis (Turcz.) Baill. fruits and Schisandra sphenanthera Rehd. et Wils. fruits) and different growth characteristics (color, shape, etc.). The application of chemical pattern recognition in the ESI-MS(n) data analysis was carried out by principal component analysis (PCA), hierarchical cluster analysis (HCA) and linear discriminant analysis (LDA). Then the antioxidant activity of different Fructus Schisandrae samples were determined by an LC-ESI-MS method and ferric reducing antioxidant power (FRAP) assay. Using the ESI-MS(n) method coupled with chemical pattern recognition analysis and correlated with the antioxidant activity evaluation, the two similar species were successfully distinguished, thus improving the therapeutic safety and effectiveness. The superior characteristics of Schisandra chinensis (Turcz.) Baill. fruits were obtained and made the selection and breeding of Chinese medicine materials more scientific. This study indicates that ESI-MS(n) is a valuable tool for the authentication of botanical origin and can also be useful for the quality control of Chinese medicinal herbs.


[Treatment of social anxiety disorder (SAD)].
Social anxiety disorder(SAD) is the most common anxiety disorder and it is a risk factor for subsequent substance abuse. The efficacy of selective serotonin reuptake inhibitors (SSRIs) and serotonin noradrenaline reuptake inhibitor(SNRI) for the treatment of SAD has been reported. Cognitive behavioral therapy(CBT) was also effective for the treatment of SAD in many studies. High rate of co-occurring SAD and alcohol use disorder (AUD) can be found in both clinical samples and in the general population. With regard to treatments for those suffering from SAD and AUD, little is known about the most effective treatment. Early interventions with social anxiety may thwart the development of maladaptive patterns of coping-related alcohol use from developing in the first place, and effectively prevent the developing of co-morbid SAD-AUDs.


Tailored nanostructured titania integrated on titanium micropillars with outstanding wicking properties.
A novel wicking material using nanostructured titania grown on high aspect ratio titanium micropillars is demonstrated. High aspect ratio titanium micropillars were micromachined from bulk titanium sheets. Nanostructured titania was then grown on the surface of titanium micropillars by oxidation in aqueous hydrogen peroxide solution followed by thermal annealing. The nanostructured titania formed has an open porous structure with a nanoscale pore diameter and wall thickness. X-ray diffraction and pole figure studies indicate the formation of anatase phase of titania and the absence of a preferred orientation in the porous film. The hybrid nanostructured titania on titanium micropillars has excellent hydrophilic properties with a water capillary speed comparable to or exceeding that of conventional wick materials commonly used in heat pipes for the thermal management of electronic devices.


Involution: apoptosis and tissue remodelling that convert the mammary gland from milk factory to a quiescent organ.
Involution of the mammary gland is an essential process that removes the milk-producing epithelial cells when they become redundant at weaning. It is a two-step process that involves the death of the secretory epithelium and its replacement by adipocytes. During the first phase, remodelling is inhibited and apoptotic cells can be seen in the lumena of the alveoli. In the second phase, apoptosis is accompanied by remodelling of the surrounding stroma and re-differentiation of the adipocytes. Considerable effort has been directed towards understanding the molecular mechanisms of the involution process and this has resulted in the identification of the principal signalling pathways involved.