Rubisco forms a lattice inside alpha-carboxysomes.
Despite the importance of microcompartments in prokaryotic biology and bioengineering, structural heterogeneity has prevented a complete understanding of their architecture, ultrastructure, and spatial organization. Here, we employ cryo-electron tomography to image α-carboxysomes, a pseudo-icosahedral microcompartment responsible for carbon fixation. We have solved a high-resolution subtomogram average of the Rubisco cargo inside the carboxysome, and determined the arrangement of the enzyme. We find that the H. neapolitanus Rubisco polymerizes in vivo, mediated by the small Rubisco subunit. These fibrils can further pack to form a lattice with six-fold pseudo-symmetry. This arrangement preserves freedom of motion and accessibility around the Rubisco active site and the binding sites for two other carboxysome proteins, CsoSCA (a carbonic anhydrase) and the disordered CsoS2, even at Rubisco concentrations exceeding 800 μM. This characterization of Rubisco cargo inside the α-carboxysome provides insight into the balance between order and disorder in microcompartment organization.


Cloning and sequencing of the peroxisomal amine oxidase gene from Hansenula polymorpha.
We have cloned the AMO gene, encoding the microbody matrix enzyme amine oxidase (EC 126.96.36.199) from the yeast Hansenula polymorpha. The gene was isolated by differential screening of a cDNA library, immunoselection, and subsequent screening of a H. polymorpha genomic library. The nucleotide sequence of a 3.6 kilobase stretch of DNA containing the amine oxidase (AMO) gene was determined. The AMO gene contains an open reading frame of 692 amino acids, with a relative molecular mass of 77,435. The 5' and 3' ends of the gene were mapped and show that the transcribed region measures 2134 nucleotides. The derived amino-acid sequence was confirmed by sequencing an internal proteolytic fragment of the purified protein. Amine oxidase contains the tripeptide sequence Ser-Arg-Leu, located 9 residues from the carboxy terminus, which may represent the topogenic signal for protein import into microbodies.


Exploring the impact of 12-hour shifts on nurse fatigue in intensive care units.
To assess 12-h shift Intensive Care Unit (ICU) nurses' fatigue and identify the associated demographic factors. Literature reveals inconsistencies as to whether 12-h shifts decrease or increase nurse fatigue levels. A cross-sectional survey of 67 ICU nurses working 12-h shifts was undertaken to determine their fatigue levels in two hospitals. The Occupational Fatigue Exhaustion/Recovery Scale (OFER), Spearman's correlation, ANOVA, t-tests, and Chi-Square were used for analyses. 57 out of 67 participants experienced low to moderate chronic fatigue; 36 of those exhibited low to moderate acute fatigue levels; 46 reported low to moderate inter-shift fatigue. Age (ρ = 0.03, r2 = -0.28), number of family dependents (ρ = 0.03, r2 = -0.27), and years of nursing experience (ρ = 0.03, r2 = -0.27) were moderately negatively correlated with acute fatigue, while frequency of exercise per week (ρ = 0.01, r2 = -0.31) was moderately negatively correlated with chronic fatigue. Hospital A had higher chronic fatigue levels than Hospital B. Age (ρ < 0.01), age group (ρ = 0.03), shift schedule (ρ = 0.02), and nursing experience (ρ = 0.03) were significantly related to the difference in chronic fatigue levels between the two hospitals. More than half of the 12-h shift ICU nurses studied in both hospitals had low to moderate fatigue levels. Age, number of family dependents, years of nursing experience, and frequency of exercise per week were identified as key factors associated with fatigue. The difference in chronic fatigue levels between hospitals suggests that implementing more support for younger and/or less experienced nurses, better strategies for retaining more experienced nurses, and fewer rotating shifts could help reduce fatigue.


Human Papillomavirus-Associated Oral Cavity Squamous Cell Carcinoma: An Entity with Distinct Morphologic and Clinical Features.
HPV-associated oral cavity squamous cell carcinoma (SCC) is not well-characterized in the literature, and also has a clinical significance that is poorly understood. We gathered a cohort of oral cavity (OC) SCC with nonkeratinizing morphology, either in the invasive or in situ carcinoma (or both), tested for p16 by immunohistochemistry and high risk HPV E6/E7 mRNA by RTPCR (reference standard for transcriptionally-active high risk HPV) and gathered detailed morphologic and clinicopathologic data. Thirteen patients from two institutions were proven to be HPV-associated by combined p16 and high risk HPV mRNA positivity. All 13 patients (100%) were males, all were heavy smokers (average 57 pack/year), and most were active drinkers (9/11 or 81.8%). All 13 (100%) involved the tongue and/or floor of mouth. All had nonkeratinizing features, but maturing squamous differentiation varied widely (0-90%; mean 37.3%). Nonkeratinizing areas had high N:C ratios and larger nests, frequently with pushing borders, and minimal (or no) stromal desmoplasia. The carcinoma in situ, when present, was Bowenoid/nonkeratinizing with cells with high N:C ratios, full thickness loss of maturation, and abundant apoptosis and mitosis. HPV was type 16 in 11 patients (84.6%) and type 33 in two (15.4%). Nine patients had treatment data available. These underwent primary surgical resection with tumors ranging from 1.6 to 5.2 cm. Most had bone invasion (6/9-66.7% were T4a tumors), and most (6/9-66.7%) had extensive SCC in situ with all 6 of these patients having final margins positive for in situ carcinoma. HPV-associated OCSCC is an uncommon entity that shows certain distinct clinical and pathologic features. Recognition of these features may help pathologic diagnosis and could potentially help guide clinical management.


Large scale survey of yeasts in frozen concentrated orange juice (FCOJ): Occurrence, diversity, and resistance to peracetic acid.
This study aimed to evaluate the occurrence and diversity of yeasts in frozen concentrated orange juice (FCOJ) and assess the resistance of yeasts to peracetic acid. One thousand five hundred (n = 1500) samples of frozen concentrated orange juice (FCOJ) were analyzed, and 280 yeast strains were isolated and identified. Candida represented 37% of all isolates, and the main species identified were Candida pseudointermedia and C. orthopsilosis. Other yeasts identified were Starmerella, Wickerhamiella, Wickerhamiella, Clavispora, Kodamaea, Meyerozyma, Rhodotorula, Trichosporon, Wickerhamomyces, Kluyveromyces, Hanseniaspora, Saccharomyces, Torulaspora, and Zygosaccharomyces. The exogenous origin of the contamination in FCOJ samples analyzed was shown by the high diversity, corroborated by the Simpson (D) and Shannon (H') indices. From a total of 227 yeasts strains tested, more than 20% were able to withstand peracetic acid concentrations >200 ppm, with emphasis on W. anomalus (300 ppm), W. sergipensis (350 ppm), C. rugopelliculosa (350 ppm), K. marxianus (450 ppm), C. parapsilosis (500 ppm), C. pseudointermedia (500 ppm), W. sorbosivorans (500 ppm), C. boleticola (600 ppm), S. cerevisiae (700 ppm) and C. orthopsilosis (750 ppm). This study adds novel data regarding the occurrence and diversity of yeasts present in FCOJ and their resistance to a chemical compound commonly employed in the sanitization of processing and distribution premises and vehicles. These findings are essential to support the development of measures for proper mitigation of contamination of orange juice towards reducing the risks of spoilage by yeasts during FCOJ transportation/storage or when FCOJ is used as an ingredient.


Flow cytometry as a new method to quantify the cellular content of human saliva and its relation to gingivitis.
Determining the cellular content of saliva by means of conventional microscopy chamber counting is a very time-consuming and operator-sensitive procedure. This study concentrated on the use of flow cytometry to examine the cellular content of saliva. Erythrocytes, leukocytes, epithelial cells and bacteria were quantified and the results were compared with caries experience and the presence of gingivitis. 258 uncentrifuged vortexed paraffin-stimulated saliva samples (112 males and 146 females) were analyzed with the UF-100 flow cytometer. Salivary reference values were established for erythrocyte, leukocyte, epithelial cell and bacterial count. Caries experience (DMF) and the presence of gingivitis were recorded. Caries experience or caries risk could not be assessed with flow cytometry. However, salivary flow cytometry may be useful in determining an individual's risk for gingivitis: a significant increase in salivary leukocytes was observed in individuals with gingivitis. At a cut-off level of 10(3) leukocytes micro l(-1) saliva, a sensitivity of 76% and a specificity of 45% was obtained. Other analytes were not significantly different between individuals with and without gingivitis. Flow cytometry of paraffin-stimulated human saliva seems a promising diagnostic or predictive tool and further investigations of diseases of the oro-pharyngeal loge, such as tonsillitis and periodontitis, should be carried out in the future.


Changes in histamine metabolism in the mouse hypothalamus induced by acute administration of ethanol.
The effect of acute ethanol administration on histamine (HA) dynamics was examined in the mouse hypothalamus. The steady-state level of HA did not change after intraperitoneal administration of ethanol (0.5-5 g/kg), whereas the level of tele-methylhistamine (t-MH), a predominant metabolite of brain HA, increased when 3 and 5 g/kg of ethanol was given. Pargyline hydrochloride (80 mg/kg, i.p.) increased the level of t-MH by 72.2% 90 min after the treatment. Ethanol at any dose given did not significantly affect the t-MH level in the pargyline-pretreated mice. Decrease in the t-MH level induced by metoprine (10 mg/kg, i.p.), an inhibitor of HA-N-methyltransferase, was suppressed by ethanol (5 g/kg), thereby suggesting inhibition of the elimination of brain t-MH. Ethanol (5 g/kg) significantly delayed the depletion of HA induced by (S)-alpha-fluoromethylhistidine (50 mg/kg, i.v.), a specific inhibitor of histidine decarboxylase. Therefore, a large dose of ethanol apparently decreases HA turnover in the mouse hypothalamus.


In vivo monitoring of caspase activation using a fluorescence resonance energy transfer-based fluorescent probe.
Caspases, which constitute a family of cysteine proteases, are highly conserved in multicellular organisms and function as a central player in apoptosis. The detection of apoptosis is intrinsically difficult because dying cells are rapidly removed from tissues by phagocytosis. Thus, the development of a method for detecting caspase activation is critical for the in vivo study of apoptosis. In this chapter, we describe a genetically encoded fluorescent probe for live imaging of caspase activation.


Characterization of antibacterial activity of a N-acetylmuramoyl-L-alanine amidase produced by Latilactobacillus sakei isolated from salami.
Lactic acid bacteria are the predominant group within meat products, whose metabolites such as bacteriocins and peptidoglycan hydrolases inhibit pathogenic or spoilage bacteria. Fermented meat products, as a salami, is a good source to analyze the viable microbiota, due to these products present a low risk to consumer health. The aim of this work was to identify the lactic acid bacteria with broad antibacterial activity present in salami, purify the protein responsible for this activity, achieve antagonistic spectrum and perform the biochemical characterization. Five strains from salami were selected, isolated and identified by 16S rRNA gene sequencing. The antimicrobial activity was evaluated by antagonism assay and zymography, using spoilage microorganisms commonly found in meat products. The strain that showed a broad antibacterial activity was Latilactobacillus sakei and the antibacterial activity was given by a protein with 75-kDa of molecular mass, identified by LC/MALDI-TOF/TOF. The sequence analysis showed 67% of identity with a N-acetylmuramoyl-L-alanine amidase protein with five non-identical LysM domains. The purified protein showed an optimal pH of 8.0 and heat resistance at 80 °C for 10 min. L. sakei strain displayed antibacterial activity against Gram-negative and Gram-positive spoilage microorganisms. The results of this study provide the information to use Latilactobacillus sakei as a starter culture which will provide the necessary metabolites to combat undesirable microorganisms. Additionally, the conditions and properties for the best application and use of the antibacterial protein produced by this strain. This protein may have a potential use in the food industry as a new antibacterial agent.


Detection of Clostridioides difficile in hospital environment by using C diff Banana Broth™.
116 environmental samples from a 504 bed clinical hospital obtained in 2017/19 were inoculated into C diff Banana Broth™. Six C. difficile and 12 C. pefringens strains were isolated. Antibiotic-resistant Clostridium spp. dominated in hospital environment. To determine Clostridium spp. in hospital environment suitable medium like C diff Banana Broth™ should be used.


Neuromonitoring After Cardiac Arrest: Can Twenty-First Century Medicine Personalize Post Cardiac Arrest Care?
Cardiac arrest survivors comprise a heterogeneous population, in which the etiology of arrest, systemic and neurologic comorbidities, and sequelae of post-cardiac arrest syndrome influence the severity of secondary brain injury. The degree of secondary neurologic injury can be modifiable and is influenced by factors that alter cerebral physiology. Neuromonitoring techniques provide tools for evaluating the evolution of physiologic variables over time. This article reviews the pathophysiology of hypoxic-ischemic brain injury, provides an overview of the neuromonitoring tools available to identify risk profiles for secondary brain injury, and highlights the importance of an individualized approach to post cardiac arrest care.


Depolarization-induced slowing of Ca2+ channel deactivation in squid neurons.
Properties of squid giant fiber lobe (GFL) Ca2+ channel deactivation (closing) were studied using whole-cell voltage clamp. Tail currents displayed biexponential decay, and fast and slow components of these tails exhibited similar external Ca(2+)- and voltage-dependence. Both components also shared similar inactivation properties. Increasing duration pulses to strongly depolarizing potentials caused a substantial slowing of the rate of deactivation for the fast component, and also led to an apparent conversion of fast tail currents to slow without an increase in total tail amplitude. A five-state kinetic model that computed the closing of channels differentially populating two open states could simulate the kinetic characteristics of GFL Ca2+ pulse and tail currents over a wide voltage range. The kinetics of the proposed state transition was very similar to the time course of relief of omega-Agatoxin IVA Ca2+ channel block with long pulses. A similar model predicted that the relief of block could occur via faster toxin dissociation from the second open state. Thus, GFL Ca2+ channels possess a unique form of voltage-dependent gating modification, in which maintained prior depolarization leads to a significant delay to channel closure at negative potentials. At the nerve terminal, amplified Ca2+ signals generated by such a mechanism might alter synaptic responses to repetitive stimulation.


[Caregiver burden: From recognition to care].
The burden of caregivers of patients with chronic diseases has been the subject of research for several years, but few studies have focused on the suffering of family members from the time of their first hospitalization in psychiatry. The identification of the determinants of this burden would allow us to propose early management aimed at improving the well-being of the patients' family and friends and maximizing intra-family resources.


Light diffraction study of single skeletal muscle fibres.
Light diffraction patterns from isolated frog semitendinosus muscle fibers were examined. When transilluminated by laser light, the muscle striations produce a diffraction pattern consisting of a series of lines that are projected as points onto an optical detector by a lens system. Diffraction data may be sequentially stored every 18 ms for later processing by digital computer systems. First- and second-order diffraction line intensities were examined from intact, chemically skinned, and glycerinated single fibers. The diffraction line intensities demonstrated a strong length dependence upon passive stretch from reference length to 3.6 micrometer. The first-order intensity linearly increased an average of 15-fold over the range examined. The magnitude of the second order intensity was less than the first order and showed an exponential rise with increasing length. Both first- and second-order intensities decreased upon muscle activation. Data from chemically skinned and glycerinated single fibers were not significantly different from intact fibers, indicating that the membrane structure has little effect upon the diffraction phenomenon in muscle. Theoretical model systems are examined in an attempt to find the basis of these results. Neither an analysis based on a diffraction grating with variable spacing nor the unit cell model of Fujime provides an explanation for the observed length dependency of intensity. Though the origin of the intensity decrease upon stimulation is not known, we have suggested that it could result from lateral misalignment of myofibrils and can occur upon activation.


Luteal phase support using either Crinone 8% or Utrogest: results of a prospective, randomized study.
The Crinone 8% preparation makes it possible to administer natural progesterone (90 mg) vaginally once daily for luteal phase support (LPS). Until now, no prospective, randomized studies have directly compared this new preparation with widely used Utrogest capsules, which were originally designed for oral administration but are used routinely as a vaginal preparation. A prospective, randomized study investigated 126 patients undergoing cycles of in vitro fertilization (IVF) and IVF/intracytoplasmic sperm injection (ICSI). Patients received either Crinone 8% (n = 73) vaginally once daily or two Utrogest capsules (n=53) vaginally three times daily (600 mg). Clinical pregnancy rates were comparable (28.8 versus 18.9%), as were clinical abortion rates until 12 weeks of gestation (14.3 versus 10.0%) and clinical ongoing pregnancy rates (24.7 versus 17.0%) in the Crinone 8% and Utrogest groups, respectively. Forty-seven non-pregnant patients were randomly selected to answer questions regarding comfort during LPS. Crinone 8% had a clear advantage over Utrogest as it resulted in less vaginal discharge (P < 0.01) and fewer application difficulties (P<0.05). Twenty patients familiar with the alternative preparation from a previous cycle also noted that Crinone 8% was easier to apply (P < 0.01) and less time consuming (P < 0.05) to use than Utrogest.


Genome Resequencing Reveals Rapid, Repeated Evolution in the Colorado Potato Beetle.
Insecticide resistance and rapid pest evolution threatens food security and the development of sustainable agricultural practices, yet the evolutionary mechanisms that allow pests to rapidly adapt to control tactics remains unclear. Here, we examine how a global super-pest, the Colorado potato beetle (CPB), Leptinotarsa decemlineata, rapidly evolves resistance to insecticides. Using whole-genome resequencing and transcriptomic data focused on its ancestral and pest range in North America, we assess evidence for three, nonmutually exclusive models of rapid evolution: pervasive selection on novel mutations, rapid regulatory evolution, and repeated selection on standing genetic variation. Population genomic analysis demonstrates that CPB is geographically structured, even among recently established pest populations. Pest populations exhibit similar levels of nucleotide diversity, relative to nonpest populations, and show evidence of recent expansion. Genome scans provide clear signatures of repeated adaptation across CPB populations, with especially strong evidence of selection on insecticide resistance genes in different populations. Analyses of gene expression show that constitutive upregulation of candidate insecticide resistance genes drives distinctive population patterns. CPB evolves insecticide resistance repeatedly across agricultural regions, leveraging similar genetic pathways but different genes, demonstrating a polygenic trait architecture for insecticide resistance that can evolve from standing genetic variation. Despite expectations, we do not find support for strong selection on novel mutations, or rapid evolution from selection on regulatory genes. These results suggest that integrated pest management practices must mitigate the evolution of polygenic resistance phenotypes among local pest populations, in order to maintain the efficacy and sustainability of novel control techniques.


High frequency of ocular toxoplasmosis in Quindío, Colombia and risk factors related to the infection.
To determine the frequency of retinochoroidal lesions by ocular toxoplasmosis and their relationships with risk factors, in residents of two districts with high exposure to Toxoplasma, in Armenia-Quindío, Colombia. Cross-sectional analyses of fundoscopy screening, serological tests, and questionnaires were performed to determine risk factors associated with ocular toxoplasmosis retinochoroidal lesions. Differences in proportions were analyzed using the chi-squared test. Of 161 individuals examined, 17 (10.5%) exhibited retinochoroidal scars suggestive of old inactive Toxoplasma gondii infection. All 17 individuals were seropositive for T. gondii antibodies. Consumption of bottled water was protective against T. gondii infection among individuals in this study. There were no specific epidemiological risk factors associated with ocular toxoplasmosis retinochoroidal lesions. Ocular toxoplasmosis is an important cause of visual impairment in Armenia-Quindío, Colombia. The consumption of boiled or bottled water is a major preventive public health measure to reduce infection by T. gondii and the subsequent onset of OT.


Combined effects of ozone and cigarette smoke on airway responsiveness and vascular permeability in guinea pigs.
The effects of combined exposure to ozone and cigarette smoke on airway responsiveness and tracheal vascular permeability, compared with those of single exposure were examined in guinea pigs. Airway responsiveness was assessed by measuring the specific airway resistance (sRaw) as a function of increasing concentration of inhaled methacholine aerosol immediately, 5 hr, and 24 hr after exposure. In a parallel study, tracheal vascular permeability was quantified by measuring the tracheal extravasation of intravenously administered Evans blue dye. Neither exposure to 1 ppm ozone for 30 min nor 5 puffs of cigarette smoke increased airway responsiveness or vascular permeability at any time after exposure. Combined exposure to 1 ppm ozone for 30 min and 5 puffs of cigarette smoke caused airway hyper-responsiveness and increased vascular permeability immediately after exposure. Exposure to 1 ppm ozone for 90 min increased both airway responsiveness and vascular permeability immediately after exposure. Exposure to 10 puffs of cigarette smoke increased airway responsiveness but not vascular permeability immediately after exposure. Combined exposure to 1 ppm ozone for 90 min and 10 puffs of cigarette smoke increased both airway responsiveness and vascular permeability immediately after exposure. The combined exposure to ozone and cigarette smoke thus increased both airway responsiveness and tracheal vascular permeability to a greater extent than did exposure to a single agent, suggesting that a combination of air pollutants has a more deleterious effect both on airway responsiveness and on tracheal vascular permeability than does either agent alone in guinea pigs.


RGD-conjugated silica-coated gold nanorods on the surface of carbon nanotubes for targeted photoacoustic imaging of gastric cancer.
Herein, we reported for the first time that RGD-conjugated silica-coated gold nanorods on the surface of multiwalled carbon nanotubes were successfully used for targeted photoacoustic imaging of in vivo gastric cancer cells. A simple strategy was used to attach covalently silica-coated gold nanorods (sGNRs) onto the surface of multiwalled carbon nanotubes (MWNTs) to fabricate a hybrid nanostructure. The cross-linked reaction occurred through the combination of carboxyl groups on the MWNTs and the amino group on the surface of sGNRs modified with a silane coupling agent. RGD peptides were conjugated with the sGNR/MWNT nanostructure; resultant RGD-conjugated sGNR/MWNT probes were investigated for their influences on viability of MGC803 and GES-1 cells. The nude mice models loaded with gastric cancer cells were prepared, the RGD-conjugated sGNR/MWNT probes were injected into gastric cancer-bearing nude mice models via the tail vein, and the nude mice were observed by an optoacoustic imaging system. Results showed that RGD-conjugated sGNR/MWNT probes showed good water solubility and low cellular toxicity, could target in vivo gastric cancer cells, and obtained strong photoacoustic imaging in the nude model. RGD-conjugated sGNR/MWNT probes will own great potential in applications such as targeted photoacoustic imaging and photothermal therapy in the near future.


Saccharide microarrays for high-throughput interrogation of glycan-protein binding interactions.
This chapter describes two methods for fabricating microarrays of saccharides for display and interrogation with binding proteins, using fluorescence detection. The first approach is based on the rapid immobilization of heparan sulphate glycans upon commercially available aminosilane slides via their reducing ends. The second approach is based on the use of a hydrazide-derivatized self-assembled monolayer (SAM) on a gold-coated slide surface. Both provide for efficient and chemoselective attachment and anchoring of oligosaccharide probes via their reducing ends, enabling the large-scale arraying of natural saccharides without cumbersome pre-derivatization. The latter platform, in particular, also has the potential for use with other biophysical readout methods including matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy, surface plasmon resonance, and quartz crystal microbalances. These microarray platforms provide a facile approach for interrogating multiple carbohydrate-protein interactions in a high-throughput manner using minimal quantities of reagents. They provide an essential new experimental strategy in the growing armoury of the glycomics toolkit.


Bioenergy from dairy manure: technologies, challenges and opportunities.
Dairy manure (DM) is a kind of cheap cellulosic biomass resource which includes lignocellulose and mineral nutrients. Random stacks not only leads damage to the environment, but also results in waste of natural resources. The traditional ways to use DM include returning it to the soil or acting as a fertilizer, which could reduce environmental pollution to some extent. However, the resource utilization rate is not high and socio-economic performance is not utilized. To expand the application of DM, more and more attention has been paid to explore its potential as bioenergy or bio-chemicals production. This article presented a comprehensive review of different types of bioenergy production from DM and provided a general overview for bioenergy production. Importantly, this paper discussed potentials of DM as candidate feedstocks not only for biogas, bioethanol, biohydrogen, microbial fuel cell, lactic acid, and fumaric acid production by microbial technology, but also for bio-oil and biochar production through apyrolysis process. Additionally, the use of manure for replacing freshwater or nutrients for algae cultivation and cellulase production were also discussed. Overall, DM could be a novel suitable material for future biorefinery. Importantly, considerable efforts and further extensive research on overcoming technical bottlenecks like pretreatment, the effective release of fermentable sugars, the absence of robust organisms for fermentation, energy balance, and life cycle assessment should be needed to develop a comprehensive biorefinery model.


Rapid characterization of the chemical constituents of Yinchen Wuling Powder by UPLC coupled with Fourier transform ion cyclotron resonance mass spectrometry.
Yinchen Wuling Powder (YCWLP) is a classic Chinese medicine prescription with a long history and has been commonly used for treating jaundice hepatitis, liver fibrosis, hyperlipidemia and early diabetes in clinical applications. However, the chemical composition of YCWLP is still unclear. In order to obtain the chemical profile of YCWLP, a systematic ultra-performance liquid chromatography coupled with fourier transform ion cyclotron resonance mass spectrometry (UPLC-FT-ICR-MS) method was developed in this study. As a result, a total of 138 compounds including terpenoid acids, organic acids, flavonoids, sesquiterpenes, coumarins and anthraquinones were identified by comparing the retention time, molecular ions and fragmentation behaviors with the reference compounds or the in-house database. This study comprehensively elucidated the chemical basis of YCWLP and provided a scientific basis for further quality control and pharmacology research.


A phosphorylcholine-modified chitosan polymer as an endothelial progenitor cell supporting matrix.
The aim of the present study was to develop a new biopolymer to increase endothelial progenitor cells (EPC) survival and amplification. As a cell culture platform, bone marrow-derived cells (BMDC) were used to investigate the biocompatibility of chitosan-phosphorylcholine (CH-PC). On CH-PC, BMDC were found in colonies with a mortality rate similar to that of fibronectin (FN), the control matrix. Adhesion/proliferation assays demonstrated a greater number of BMDC on CH-PC after 7 days with an amplification phase occurring during the second week. Difference in adhesion mechanisms between (CH-PC) and the control FN matrix suggest distinctive cell retention ability. Confocal microscopy analyses confirmed that (CH-PC) supported the survival/differentiation of endothelial cells. Moreover, flow cytometry analyses demonstrated that, (CH-PC) increased the percentage of progenitor cells (CD117(+)CD34(+)) (7.1 ± 0.8%, FN: 4.1 ± 0.8%) and EPC (CD117(+)CD34(+)VEGFR-2(+)CD31(+)) (2.33 ± 0.6%, FN: 0.25 ± 0.1%), while the mesenchymal stem cell fraction (CD44(+)CD106(+)CD90(+)) was decreased (0.07 ± 0.01%, FN: 0.55 ± 0.22%). Polymeric substrate CH-PC might provide a suitable surface to promote the amplification of EPC for future vascular therapeutic applications.


Long-term stability of the oxygen pulse curve during maximal exercise.
Exercise oxygen pulse (O₂ pulse), a surrogate for stroke volume and arteriovenous oxygen difference, has emerged as an important variable obtained during cardiopulmonary exercise testing. We hypothesized that the O₂ pulse curve pattern response to a maximal cycling ramp protocol exhibits a stable linear pattern in subjects reevaluated under the same clinical conditions. We retrospectively studied 100 adults (80 males), mean age at baseline of 59 + 12 years, who performed two cardiopulmonary exercise testings (median interval was 15 months), for clinical and/or exercise prescription reasons. The relative O₂ pulse was calculated by dividing its absolute value by body weight. Subjects were classified into quintiles of relative O₂ pulse. Cardiopulmonary exercise testing results and the O₂ pulse curve pattern, expressed by its slope and intercept, were compared among quintiles of relative O₂ pulse at both cardiopulmonary exercise testings. After excluding the first minute of CPX (rest-exercise transition), the relative O₂ pulse curve exhibited a linear increase, as demonstrated by high coefficients of determination (R² from 0.75 to 0.90; p < 0.05 for all quintiles). Even though maximum oxygen uptake and relative O₂ pulse were significantly higher in the second cardiopulmonary exercise testing for each quintile of relative O₂ pulse (p < 0.05 for all comparisons), no differences were found when slopes and intercepts were compared between the first and second cardiopulmonary exercise testings (p > 0.05 for all comparisons; except for intercept in the 5th quintile). Excluding the rest-exercise transition, the relative O₂ pulse exhibited a stable linear increase throughout maximal exercise in adults that were retested under same clinical conditions.


First isolation and molecular characterization of canine parvovirus-type 2b (CPV-2b) from red foxes (Vulpes vulpes) living in the wild habitat of Turkey.
The canine parvovirus, with its many variants, is responsible for a pivotal and common viral infection affecting millions of dogs and other carnivore species worldwide, particularly the wild ones, which are considered as the main reservoir hosts. To that end, this study investigated the presence of canine parvovirus (CPV) in red foxes (Vulpes vulpes) living in wild habitats of several regions of Turkey. We randomly collected 630 archival fox stool specimens from rural areas of 22 provinces and used real-time PCR to detect CPV. Two of the 630 (0.3%) stool samples were positive for CPV-DNA, named Tr-Fox/128(Aydın) and Tr-Fox/159(Manisa). We attempted to isolate the virus in a MDCK cell line, and cytopathic effects were observed four days post-inoculation. Three regions corresponding to the CPV capsid protein VP2 gene from extracted DNA of positive samples were amplified by conventional PCR, and the products were visualised, purified, and Sanger sequenced. Three overlapping DNA raw sequence fragments, were read, assembled, and aligned to obtain approximately 1.5 kb-long regions that cover most of the VP2 gene, then deposited in GenBank. After comparing the isolates with parvovirus sequences data of domestic and wild carnivores by BLAST processing, our isolates' similarity rate with each other was 99.40%, with base differences in 9 nucleotide positions. They were classified as 2b variant closely related to isolates from dogs in Turkey, Egypt, Iraq, Italy, Thailand, and China. This study presents evidence of interspecies transmission of CPV, of which there are no reports on prevalence in wildlife carnivores of our country. Identification of CPV in red foxes threatens local and hunting dogs, which may contract the infection or disseminate it to other wild animal species or vice-versa.


Hallucinations and Delusions as Low-Quality Attributions: Influencing Factors and Proposal for Their Analysis.
Hallucinations and delusions, in keeping with the distress accompanying them, are major features in the diagnosis of psychosis in international classifications. In spite of their human and clinical importance, the concepts are unclear. The distinction between hallucinations and delusions in terms of perception-thought is not precise enough, causing problems in analyzing the patient's words. Nor are the differentiations or variations within each precise enough. Continuing the long clinical tradition discussing the distinction between hallucinations and delusions while assuming their similarities, this study poses a concept integrating the two phenomena as attributions people make about themselves and their settings. Then the elements of any attribution can be used as guides for structuring significant literature on both, and reduce analytical ambiguity. Such attributions make more sense within the structure of two-way relationships with factors in a person's own framework and setting. This structure is described with its variables and relationships as a guide to assessment, follow-up, and intervention. Two checklists are provided for orientation.


Correlation of Polymorphonuclear Cell Burden and Microbial Growth to the Inflammatory Cytokines in Tracheal Aspirates from Ventilated Preterm Infants.
The significance of the presence of microorganisms and polymorphonuclear cells in the tracheal aspirates (TAs) of ventilated preterm infants is not well known. Our aim was to correlate information about the presence of polymorphonuclear cells with microbial growth and the cytokine milieu in the TAs of infants who have been intubated for >7 days. TAs were collected from infants who had been intubated for 7 days or longer. Respiratory cultures were performed, and infants were stratified based on the presence and abundance of polymorphonuclear cells and microbial growth. Cytokines were measured in the TAs of each of the respective groups. In the 19 infants whose TAs were collected, the presence of at least moderate WBC with presence of microbial growth was positively associated with the presence of interleukin (IL)-10, IL-1β, IL-8, and tumor necrosis factor (TNF)-α. The presence of at least moderate WBC, with or without microbial growth, was correlated positively with the presence of IL-8 and TNF-α. There are higher levels of proinflammatory cytokines (especially, IL-10, IL-1β, and TNF-α) in TAs with higher cell counts and presence of microbial growth. The findings suggest that the presence of microbial growth correlated with inflammatory burden and warrant a larger study to see if treatment of microbial growth can ameliorate the inflammatory burden. · Concomitant evaluation of inflammatory cells, microbial growth, and cytokines in tracheal aspirates.. · Moderate TA WBC with presence of microbial growth associated with IL-10, IL-1β, IL-8, and TNF-α.. · Moderate TA WBC, with/without microbial growth, correlated with the presence of IL-8 and TNF-α.. · Higher levels of IL-10, IL-1β, and TNF-α correlated with higher TA cell counts and microbial growth..


Laser scanning cytometry: principles and applications.
The laser scanning cytometer (LSC) is the microscope-based cytofluorometer that offers a plethora of analytical capabilities. Multilaser-excited fluorescence emitted from individual cells is measured at several wavelength ranges, rapidly (up to 5000 cells/min), with high sensitivity and accuracy. The following applications of LSC are reviewed: (1) identification of cells that differ in degree of chromatin condensation (e.g., mitotic or apoptotic cells or lymphocytes vs granulocytes vs monocytes); (2) detection of translocation between cytoplasm vs nucleus or nucleoplasm vs nucleolus of regulatory molecules such as NF-kappaB, p53, or Bax; (3) semiautomatic scoring of micronuclei in mutagenicity assays; (4) analysis of fluorescence in situ hybridization; (5) enumeration and morphometry of nucleoli; (6) analysis of phenotype of progeny of individual cells in clonogenicity assay; (7) cell immunophenotyping; (8) visual examination, imaging, or sequential analysis of the cells measured earlier upon their relocation, using different probes; (9) in situ enzyme kinetics and other time-resolved processes; (10) analysis of tissue section architecture; (11) application for hypocellular samples (needle aspirate, spinal fluid, etc.); (12) other clinical applications. Advantages and limitations of LSC are discussed and compared with flow cytometry.


Nonsense-mediated mRNA decay (NMD) in animal embryogenesis: to die or not to die, that is the question.
Nonsense-mediated mRNA decay (NMD) is a well-studied cellular quality-control pathway. It decreases the half-lives of eukaryotic mRNAs that aberrantly contain premature termination codons and additionally regulates an estimated 10-20% of normal transcripts. NMD factors play crucial roles during embryogenesis in many animals. Here, we review data indicating that NMD factors are required for proper embryogenesis by discussing the abnormal developmental phenotypes that result when the abundance of individual NMD factors is either downregulated or completely eliminated. We conclude that while NMD per se affects the embryogenesis of all animals, it is required to avoid embryonic lethality in only some animals. The critical roles of many NMD factors in other metabolic pathways undoubtedly also contribute to embryonic development if not viability.


Cat scratch disease bacilli in the conjunctiva of patients with Parinaud's oculoglandular syndrome.
Unilateral conjunctivitis with regional lymphadenitis has been designated Parinaud's oculoglandular syndrome (POGS). The cause has been attributed to various infectious agents, especially leptothrix and the unidentified agent of cat scratch disease (CSD). In 1983-84 the cause of CSD in lymph node and skin was established as small pleomorphic gram-negative bacilli. We now report identical bacteria in the conjunctival lesions of 9 of 24 patients with POGS. This confirms the belief that the CSD bacillus is a major cause of POGS.


Disseminated histoplasmosis in a heart transplant recipient from Saudi Arabia: A case report.
Histoplasma is endemic in North and Central America. We describe a case of disseminated histoplasmosis in a heart transplant recipient outside the known endemic areas. A 68-year-old gentleman known to have dilated cardiomyopathy. He underwent left ventricular assist device (LVAD) implantation in India and 2 years later did heart transplant in King Faisal Specialist and Research Center Hospital. Six weeks post-transplant he presented with headache and fever. All investigations were negative, and he was discharged home. Four days after discharge he presented with headache, fever, blurred vision, and an episode of loss of consciousness. Examination showed an ill looking patient who is highly febrile. Repeated work up showed pancytopenia. A repeat LP was negative. Bone marrow biopsy showed Small intracellular organisms. Extended work up revealed a positive Histoplasma urinary antigen, positive Histoplasma PCR from the bone marrow biopsy. Patient was started on Liposomal Amphotericin followed by Itraconazole with marker clinical improvement. This is the first reported case of disseminated Histoplasmosis in Saudi Arabia. We postulate that the patient had reactivation of a latent infection acquired at the time of LVAD insertion in India rather than donor derived infection by the negative fungal culture and PCR done on the donor's lung granuloma tissue.


Chaperone gp96-independent inhibition of endotoxin response by chaperone-based peptide inhibitors.
HSP90 chaperones a large number of proteins, and it plays essential roles in multiple signaling pathways to maintain protein homeostasis in the cytosol. In addition, HSP90 has been implicated in mediating recognition of lipopolysaccharide (LPS). However, no pharmacologic agents have been developed to interrogate this pathway. Herein we demonstrate that a peptide-based inhibitor that was previously reported to inhibit the master Toll-like receptor-chaperone gp96, an endoplasmic reticulum paralog of HSP90, in fact blocks HSP90-LPS interaction. It inhibited the binding of LPS to the cell surface of both wild type and gp96-null cells and thereby abrogated the cellular response to LPS but not to other Toll-like receptor ligands. We also generated a series of peptide derivatives (named peptide inhibitors of endotoxin responsiveness (PIERs)) from the N-terminal helix structure of HSP90 and demonstrated their effectiveness in blocking LPS activity. PIER inhibition of LPS signaling was partially reversed by CD14 expression. Moreover, we found that a cell-permeable PIER abrogated HSP90 function and caused degradation of multiple known HSP90 client proteins in cancer cells. Thus, targeting HSP90 is a promising modality for treatment of both LPS-mediated pathology and cancer.


Reply to Twycross et al. Comment on "Ramos-Rincon et al. Palliative Sedation in COVID-19 End-of-Life Care. Retrospective Cohort Study. Medicina 2021, 57, 873".
First of all, we want to thank Twycross, Wong, and Vivat [...].


Primary structure of scorpion anti-insect toxins isolated from the venom of Leiurus quinquestriatus quinquestriatus.
The amino acid sequences of insect-selective scorpion toxins, purified from the venom of Leiurus quinquestriatus quinquestriatus, have been determined by automatic phenyl isothiocyanate degradation of the S-carboxymethylated proteins and derived proteolytic peptides. The excitatory toxin Lqq IT1 and Lqq IT1' (70 residues) show the shift of one half-cystine from an external position, which is characteristic of anti-mammal toxins, to an internal sequence position. Lqq IT2 (61 residues) displays the half-cystine residue in position 12, common to the sequence of all known anti-mammal toxins; it induces flaccid paralysis on insects but is non-toxic for the mouse. Lqq IT2 structurally defines a new type of anti-insect toxins from scorpion venoms. CD spectra and immunological data are in agreement with this finding.


PTPS Facilitates Compartmentalized LTBP1 S-Nitrosylation and Promotes Tumor Growth under Hypoxia.
GTP cyclohydrolase I (GTPCH), 6-pyruvoyltetrahydropterin synthase (PTPS), and sepiapterin reductase (SR) are sequentially responsible for de novo synthesis of tetrahydrobiopterin (BH4), a known co-factor for nitric oxide synthase (NOS). The implication of BH4-biosynthesis process in tumorigenesis remains to be investigated. Here, we show that PTPS, which is highly expressed in early-stage colorectal cancer, is phosphorylated at Thr 58 by AMPK under hypoxia; this phosphorylation promotes PTPS binding to LTBP1 and subsequently drives iNOS-mediated LTBP1 S-nitrosylation through proximal-coupling BH4 production within the PTPS/iNOS/LTBP1 complex. In turn, LTBP1 S-nitrosylation results in proteasome-dependent LTBP1 protein degradation, revealing an inverse relationship between PTPS pT58 and LTBP1 stability. Physiologically, the repressive effect of PTPS on LTBP1 leads to impaired transforming growth factor β (TGF-β) secretion and thereby maintains tumor cell growth under hypoxia. Our findings illustrate a molecular mechanism underlying the regulation of LTBP1-TGF-β signaling by the BH4-biosynthesis pathway and highlight the specific requirement of PTPS for tumor growth.


Elaborate ligand-based pharmacophore exploration and QSAR analysis guide the synthesis of novel pyridinium-based potent beta-secretase inhibitory leads.
Beta-secretase (BACE) inhibitors have potential as anti-Alzheimer's disease treatments prompting us to explore the pharmacophoric space of 129 known BACE inhibitors. QSAR analysis was employed to select optimal combination of pharmacophoric models and 2D physicochemical descriptors capable of explaining bioactivity variation (r(2)=0.88, F=60.48, r(LOO)(2)=0.85, r(PRESS)(2) against 25 external test inhibitors=0.71). We were obliged to use ligand efficiency as the response variable because the logarithmic transformation of bioactivities failed to access self-consistent QSAR models. Three pharmacophoric models emerged in the successful QSAR equation suggesting at least three binding modes accessible to ligands within BACE binding pocket. QSAR equation and pharmacophoric models were validated through ROC curves and were employed to guide synthesis of novel pyridinium-based BACE inhibitors. The best inhibitor illustrated an IC(50) value of 1.0 microM against BACE.


Recent advances in treatment strategies for atopic dermatitis.
A wide range of different therapeutic regimens are used for atopic dermatitis. Although many treatment modalities are well established worldwide among clinicians, only the minority of these therapy recommendations are based on results of randomised controlled trials (RCTs). To close the gap between such 'generally' recommended therapies and therapies that are based on data from controlled trials, this review focuses not only on the pharmacological and clinical aspects of the currently proven agents, but also on the advantages and disadvantages of therapies that have not yet been completely tested.A review of the available literature concerning the pharmacological profile and also the level of evidence of therapeutic efficacy of all currently known topical and systemic agents for the treatment of atopic dermatitis reveals a large gap between the knowledge concerning the pharmacological action in vitro and the evidence of clinical efficacy in many cases. We agree with the conclusion of previous reviews that numerous therapies for atopic dermatitis urgently require more independent RCTs and especially comparative trials (e.g. corticosteroids vs calcineurin inhibitors). These are required in order to facilitate the choice of therapeutic strategy for the individual treatment of atopic dermatitis, with its broad spectrum of clinical manifestations and potential complications in adult patients and, particularly, in children.Finally, we also review preclinical trials with several new drugs. Immunomodulators appear to promise a new dimension for the future of therapy for atopic dermatitis, especially for severe and otherwise refractory forms or as alternatives to corticosteroids, that is, to treat facial atopic eczema without the risk of adverse effects.


Contributions of subdiaphragmatic activity, attenuation, and diaphragmatic motion to inferior wall artifact in attenuation-corrected Tc-99m myocardial perfusion SPECT.
Subdiaphragmatic activity and diaphragmatic motion both contribute to inferior wall artifacts in technetium 99m myocardial perfusion single photon emission computed tomography (SPECT). We used an anthropomorphic phantom with ventricular wall activity, liver/spleen inserts containing variable Tc-99m activity, and variable vertical (diaphragmatic) motion amplitude. SPECT and transmission scans were obtained on a GE Optima NX camera. Data were processed by use of filtered backprojection or attenuation correction (AC). Resulting myocardial activity maps were analyzed with standardized inferior-anterior and anterior-lateral wall ratios. At a subdiaphragmatic-myocardial activity ratio of 0.5:1, inferior wall attenuation predominates, producing a cold artifact. AC corrects inferior wall activity to the level of the anterior wall irrespective of diaphragmatic motion. At a subdiaphragmatic-myocardial activity ratio of 1:1, inferior wall counts vary widely depending on the proximity of subdiaphragmatic activity to the ventricle. With increasing diaphragmatic amplitude, the overlap of subdiaphragmatic activity and inferior wall worsens, leading to a complex mixture of cold and hot artifacts, not corrected by AC. Concentration and proximity of subdiaphragmatic Tc-99m activity relative to myocardium comprise a major factor in the nature and severity of inferior wall artifacts. If the subdiaphragmatic Tc-99m concentration is equivalent to that in the myocardium, complex, potentially uninterpretable hot and cold inferior wall artifacts are produced.


Role of ZBTB7A zinc finger in tumorigenesis and metastasis.
The zinc finger and BTB (broad-complex, tramtrack and bric a brac) domain containing protein 7A (ZBTB7A) is a pleiotropic transcription factor that plays an important role in various stages of cell proliferation, differentiation, and other developmental processes. ZBTB7A is a member of the POK family that directly and specifically binds to short DNA recognition sites located near their target genes thereby acting as transcriptional activator or repressor. ZBTB7A overexpression has been associated with tumorigenesis and metastasis in various human cancer types, including breast, prostate, lung, ovarian, and colon cancer. However in some instances downregulation of ZBTB7A results in tumor progression, suggesting its role as a tumor suppressor. ZBTB7A is involved with complicated regulatory networks which include protein-protein and protein-nucleic acid interactions. ZBTB7A involvement in cancer progression and metastasis is perhaps enabled through the regulation of various signaling pathways depending on the type and genetic context of cancer. The association of ZBTB7A with other proteins affects cancer aggressiveness, therapeutic resistance and clinical outcome. This review focuses on the involvement of ZBTB7A in various signaling pathways and its role in cancer progression. We will also review the literature on ZBTB7A and cancer which could be potentially explored for its therapeutic implications.


Basic neural transplantation techniques. I. Dissociated cell suspension grafts of embryonic ventral mesencephalon in the adult rat brain.
Lesions and grafts in the nigrostriatal dopamine system in rats is widely used as a model of degeneration and regeneration in the CNS, and the development for alternative strategies for treatment in Parkinson's disease. The methods of preparing a dissociated cell suspension of embryonic rat substantia nigra and its transplantation into the brain of adult rats are described. This is the first of a series of methodological reports on the basic methodology and refinements of neural transplantation techniques in the mammalian central nervous system.


Surgical treatment of popliteal artery entrapment syndrome: a ten-year experience.
Popliteal artery entrapment syndrome is increasingly described in the world literature as a cause of lower limb arterial impairment. It is caused by the anomalous interrelationship between the popliteal artery and its surrounding muscular and/or tendineous structures. The first case surgically treated was reported in 1959 and since then more than 300 cases have been reported including our personal experience (31 cases in 23 patients). We have treated surgically 19 males and four females with symptoms which were moderate (cramping after intensive physical training, paraesthesia, etc.) in 14 limbs, intermittent claudication in 16 and necrosis (first toe) in one. Preoperative arteriography showed arterial occlusion in eight limbs, stenosis in eight and aneurysms in two. In 11 limbs stenosis or occlusion was only shown after active plantar hyperextension and in two arteriography was not done because surgical indications were established on the basis of a venogram positive for popliteal vein entrapment syndrome. Ten different anatomical variants were seen and the medial head of gastrocnemius muscle was involved in 74.2%. Surgical treatment consisted of division of the aberrant musculotendinous tissue in 18 cases (in two of these balloon angioplasty was also used). In 12 cases a vascular reconstruction was also required, while one case was explored without a specific procedure being warranted. Optimal results were obtained when the syndrome was treated at an early stage by simple division of musculotendinous tissue (94.4% long-term patency rate, mean follow-up 46.0 months, min 2, max 120 months). When arterial grafting was required the long-term patency rate was only 58.3% (mean follow-up 43.5 months, min 1, max 100 months).(ABSTRACT TRUNCATED AT 250 WORDS)


Network pharmacology-based prediction of active compounds and molecular targets in Yijin-Tang acting on hyperlipidaemia and atherosclerosis.
Yijin-Tang (YJT) is a traditional prescription for the treatment of hyperlipidaemia, atherosclerosis and other ailments related to dampness phlegm, a typical pathological symptom of abnormal body fluid metabolism in Traditional Korean Medicine. However, a holistic network pharmacology approach to understanding the therapeutic mechanisms underlying hyperlipidaemia and atherosclerosis has not been pursued. To examine the network pharmacological potential effects of YJT on hyperlipidaemia and atherosclerosis, we analysed components, performed target prediction and network analysis, and investigated interacting pathways using a network pharmacology approach. Information on compounds in herbal medicines was obtained from public databases, and oral bioavailability and drug-likeness was screened using absorption, distribution, metabolism, and excretion (ADME) criteria. Correlations between compounds and genes were linked using the STITCH database, and genes related to hyperlipidaemia and atherosclerosis were gathered using the GeneCards database. Human genes were identified and subjected to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Network analysis identified 447 compounds in five herbal medicines that were subjected to ADME screening, and 21 compounds and 57 genes formed the main pathways linked to hyperlipidaemia and atherosclerosis. Among them, 10 compounds (naringenin, nobiletin, hesperidin, galangin, glycyrrhizin, homogentisic acid, stigmasterol, 6-gingerol, quercetin and glabridin) were linked to more than four genes, and are bioactive compounds and key chemicals. Core genes in this network were CASP3, CYP1A1, CYP1A2, MMP2 and MMP9. The compound-target gene network revealed close interactions between multiple components and multiple targets, and facilitates a better understanding of the potential therapeutic effects of YJT. Pharmacological network analysis can help to explain the potential effects of YJT for treating dampness phlegm-related diseases such as hyperlipidaemia and atherosclerosis.


Elevated amygdala activity during reappraisal anticipation predicts anxiety in avoidant personality disorder.
Avoidant personality disorder is characterized by pervasive anxiety, fear of criticism, disapproval, and rejection, particularly in anticipation of exposure to social situations. An important but underexplored question concerns whether anxiety in avoidant patients is associated with an impaired ability to engage emotion regulatory strategies in anticipation of and during appraisal of negative social stimuli. We examined the use of an adaptive emotion regulation strategy, cognitive reappraisal, in avoidant patients. In addition to assessing individual differences in state and trait anxiety levels, self-reported affect as well as measures of neural activity were compared between 17 avoidant patients and 21 healthy control participants both in anticipation of and during performance of a reappraisal task. Avoidant patients showed greater state and trait-related anxiety relative to healthy participants. In addition, relative to healthy participants, avoidant patients showed pronounced amygdala hyper-reactivity during reappraisal anticipation, and this hyper-reactivity effect was positively associated with increasing self-reported anxiety levels. Our finding of exaggerated amygdala activity during reappraisal anticipation could reflect anxiety about the impending need to reappraise, anxiety about the certainty of an upcoming negative image, or anxiety relating to anticipated scrutiny of task responses by the experimenters. While we believe that all of these possibilities are consistent with the phenomenology of avoidant personality disorder, future research may clarify this ambiguity. These results suggest that amygdala reactivity in anticipation of receiving negative social information may represent a key component of the neural mechanisms underlying the heightened anxiety present in avoidant patients.


Structural mechanisms of GABAA receptor autoimmune encephalitis.
Autoantibodies targeting neuronal membrane proteins can cause encephalitis, seizures, and severe behavioral abnormalities. While antibodies for several neuronal targets have been identified, structural details on how they regulate function are unknown. Here we determined cryo-electron microscopy structures of antibodies derived from an encephalitis patient bound to the γ-aminobutyric acid type A (GABAA) receptor. These antibodies induced severe encephalitis by directly inhibiting GABAA function, resulting in nervous-system hyperexcitability. The structures reveal mechanisms of GABAA inhibition and pathology. One antibody directly competes with a neurotransmitter and locks the receptor in a resting-like state. The second antibody targets the subunit interface involved in binding benzodiazepines and antagonizes diazepam potentiation. We identify key residues in these antibodies involved in specificity and affinity and confirm structure-based hypotheses for functional effects using electrophysiology. Together these studies define mechanisms of direct functional antagonism of neurotransmission underlying autoimmune encephalitis in a human patient.


The Relationship between Health Behavior and General Health Status: Based on 2011 Korea National Health and Nutrition Examination Survey.
The aim of the present study is to investigate the relationship between health behavior and general health status. We used data from the 2011 Korea National Health and Nutrition Examination Survey. Mental health was measured by stress recognition and depression. Dietary habit was measured by mixed grain diet. Life pattern was measured by sleeping time and working pattern. Physical activity was measured by walking and exercise. We defined general health status as Euro Quality of Life-5 Dimension (EQ-5Dindex), Euro Quality of Life Visual Analogue Scale (EQ-5Dvas), number of people experienced lying in a sickbed for the last one month, number of days lying in a sickbed for the last one month, and activity limitations. Mental health, dietary habit, life pattern, and physical activity have seven factors. Most of the factors have a significant correlation with EQ-5Dindex, EQ-5Dvas, number of people experienced lying in a sickbed for the last one month, number of days lying in a sickbed for the last one month, and activity limitations. Health behavior and general health status have a positive correlation.


Angiographic diagnosis and control of postoperative bleeding.
The use of diagnostic and therapeutic angiography for postoperative bleeding which began with its application for bleeding following GI surgery can be ezpanded to almost all other areas of the body. Severe postoperative hemorrhage that previously required a second operation can now be successfully managed by the use of intraarterial or intravenous vasoconstrictors or transcatheter occlusion, thus significantly reducing patient morbidity and mortality. In those patients where a reexploration becomes necessary, diagnostic angiogarphy is a useful guide and helps to tailor the operative procedures.


Structure-activity relationships for N6-substituted adenosines at a brain A1-adenosine receptor with a comparison to an A2-adenosine receptor regulating coronary blood flow.
A series of 145 N6-substituted adenosines have been screened as inhibitors of the binding of [3H]cyclohexyladenosine to an A1-adenosine receptor in rat brain membranes and the results compared to the potencies of these analogs in increasing coronary blood flow via activation of an A2-adenosine receptor. The A1 receptor shows greater stereoselectivity in the N6 region of the receptor towards asymmetric aralkyl substituents, and shows greater bulk tolerance in the N6 region of the receptor such that it retains affinity for certain N6-tertiary alkyladenosines and N6-cycloalkyladenosines that are inactive at the coronary A2 receptor. At the A1 receptor, the most potent analogs have either aliphatic N6-substituents with four or more methylene residues or have an N6-halophenyl substituent. At the A2 receptor, the most potent analogs have an N6-phenethyl or similar heteroarylethyl substituent. Certain sets or series of analogs appear useful for identifying the subtypes of adenosine receptors involved in physiological functions.


Immunological variables and acute-phase reactants in patients with ankylosing spondylitis (Bechterew's syndrome) and their relatives.
Serum and EDTA blood from 120 patients with ankylosing spondylitis (Bechterew's syndrome) and serum from 138 first-degree relatives of patients and from 42 adult blood donors were investigated. Increased serum concentrations of IgA and IgG and complement factors C3 and C4 were found in total groups of HLA B27-positive male or female patients compared with controls or relatives. The men had higher serum concentration of IgA and complement factors than the women, whereas IgM concentration was higher in the women. These patterns were found in controls, in relatives and in patients. Increased concentrations of IgA and of C4 were characteristic of all patients whereas IgG and IgM and C3 concentrations were likewise elevated in patients with peripheral joint arthritis/arthropathy. Increased levels of circulating immune complexes (CIC) were associated with peripheral joint arthritis and were strongly correlated with IgG or CRP concentrations in serum. Total haemolytic complement activity in serum was negatively correlated with concentrations of CIC or CRP indicating complement activation in patients with such complexes. No differences in serum concentrations of Ig or complement factor concentrations were seen between HLA B27-positive and negative relatives with normal sacro-iliac joints or between relatives and controls. Strong mutual correlations were seen among IgG, IgM, complement factors, CRP, SAA, sedimentation rate and alpha 2-macroglobulin. When the present findings were combined with our previous results it turned out that AS, and psoriasis with or without arthropathy, and acute anterior uveitis (AAU) in combination with sacro-iliitis, may be described as IgA-related conditions and that increased serum C4 was related to sacro-iliitis in all these disorders.


Comparison of SHARP signal system and Southern blot hybridization analysis for detection of cytomegalovirus in clinical specimens by PCR.
Thirty cytomegalovirus cell culture-positive samples were tested by the SHARP Signal System. Twenty-seven specimens (100% agreement) were identified by both methods. The SHARP Signal System is rapid (4 h), easy to perform, and potentially adaptable to automation.


Global pattern and determinants of coronavirus disease 2019 (COVID-19) vaccine coverage and progression: a global ecological study.
Understanding and minimizing existing global coronavirus disease 2019 (COVID-19) vaccination disparities is critical to global population health and eliminating health inequities. The study aims to investigate the disparities of vaccination coverage and progression and the associated economic and educational determinants to inform global COVID-19 vaccination strategies. COVID-19 vaccination coverage data from 206 countries used in the study were derived from "Our World in Data" website. After obtaining the vaccination coverage indicators, we fitted the progression indicators for vaccination. Correlation and multiple linear regression analysis were used to examine the effects of gross domestic product (GDP) per capita, Gini index, education, and their interactions on the coverage and progression of the COVID-19 vaccination. The coverage of COVID-19 vaccination ranged from less than 30 doses to more than 150 doses per hundred people, from less than 15% to more than 75% for proportion of people vaccinated, from less than 15% to more than 60% for proportion of people fully vaccinated. Similarly, the progression of vaccination ranged from less than 0.1 to more than 0.6 for progression of total number of doses, from less than 0.1 to more than 0.3 for progression of proportion of people vaccinated, and from less than 0.1 to more than 0.4 for progression of proportion of people fully vaccinated. GDP per capita and education were positively associated with the coverage and progression, while Gini index was negatively associated with the coverage and progression. Negative interaction between GDP per capita and education was also observed for coverage (β = -0.012 to -0.011, P < 0.05) and progression (β = -0.012 to -0.011, P < 0.05). Substantial geographic disparities existed for the coverage and progression of COVID-19 vaccination. Economy and education are two important factors contributing to the disparities. Different countries may adopt varied strategies to promote the national distribution and vaccination of COVID-19 vaccines.


The Effects of Chlorhexidine Dressing on Health Care-Associated Infection in Hospitalized Patients: A Meta-Analysis.
To assess the effects of chlorhexidine dressing on health care-associated infection in hospitalized patients. We searched for English-language published randomized controlled trials (RCTs) in Cochrane Library, EMBASE and PubMed between January 1998 and January 2018. We used meta-analysis to calculate the risk ratios (RRs) and 95% confidence intervals (CIs) of the data, and using the I 2 assessment to summarize the heterogeneity of RCTs and the funnel plot and Egger regression test to evaluate publication bias. A total of 13 RCTs were included in our meta-analysis, including 7555 patients and 11,931 catheters. The effects of chlorhexidine dressing on the incidence of catheter-related bloodstream infections (CRBSIs) were reported in 13 RCTs, and the incidence of CRBSIs were 1.3% (80/6160) in the chlorhexidine group and 2.5% (145/5771) in the control group. We used a forest plot to determine the risk ratio (RR) of chlorhexidine dressing on the incidence of CRBSIs, and our results showed that chlorhexidine dressing significantly reduced the incidence of CRBSIs (RR 0.55, 95% CI 0.39-0.77, P<0.001). Moreover, we also analyzed the effects of chlorhexidine dressing on the incidence of catheter colonization and catheter-related infections (CRIs), and our forest plot results showed that chlorhexidine dressing significantly reduced the incidence of catheter colonization (RR 0.52, 95% CI 0.40-0.67, P<0.001) and the incidence of CRIs (RR 0.43, 95% CI 0.28-0.66, P<0.001) in hospitalized patients. The use of chlorhexidine dressings for hospitalized patients significantly reduce the incidence of CRBSIs, catheter colonization and CRIs.


Androgen- and estrogen-receptor mediated activities of 4-hydroxytestosterone, 4-hydroxyandrostenedione and their human metabolites in yeast based assays.
4-Hydroxyandrost-4-ene-3,17-dione, also named formestane, is an irreversible aromatase inhibitor and therapeutically used as anti-breast cancer medication in post-menopausal women. Currently, no therapeutical indication led to approval of its 17-hydroxylated analog 4-hydroxytestosterone, an anabolic steroid. However, it is currently investigated in a clinical trial for breast cancer. In context with sports doping, aromatase inhibitors are administered to reduce estrogenic side effects of misused anabolic substances or their metabolites. Therefore, both substances are prohibited in sports by the World Anti-Doping Agency (WADA). Analysis of urinary phase I and phase II metabolites showed similar results for both compounds. In the current investigation, 4-hydroxyandrost-4-ene-3,17-dione, 4-hydroxytestosterone and seven of their described urinary metabolites as well as 2α-hydroxyandrostenedione were tested in the yeast androgen screen and the yeast estrogen screen. Androgenic effects were observed for all tested substances, except for one, which showed anti-androgenic properties. With regard to the yeast estrogen screen, estrogenic effects were observed for only two metabolites at rather high concentrations, while six out of the ten substances tested showed anti-estrogenic properties. In terms of the strong androgenic effect observed for 4-hydroxytestosterone (10-8 M), 4-hydroxyandrost-4-ene-3,17-dione (10-8 M) and two more urinary metabolites, the yeast androgen assay may also be used to trace abuse in urine samples.


Femoral artery catheterization in neonates and infants.
To determine the incidence of perfusion-related complications associated with indwelling femoral artery monitoring catheters in neonates and infants following introduction of a 2.5-F diameter, 5-cm length, polyethylene catheter (Cook Medical, Bloomington, IN) to our unit. Prospective observational cohort study. Pediatric cardiac intensive care unit in a university-affiliated children's hospital. All patients <2 yrs old with an indwelling femoral artery catheter during a 3-yr period. None. Two hundred eighty-two patients (including 98 neonates), median (range) age 10 wks (0.1-84), weight 4.1 kg (2.0-11.1) were enrolled; outcomes in 249 were evaluable. Pulse strength in dorsalis pedis arteries and pulse discrepancies between feet were assessed hourly by the cardiac intensive care unit nurse and recorded on a flow sheet. Nonpalpable pulses were assessed as "absent" or "present" with ultrasonic Doppler. Following removal of the catheter, assessments of pulse strength continued until resolution of any discrepancies. Median (range) duration of catheterization was 4 days (1-23). Catheters of 2.5-F diameter were used in 227 patients and larger catheters in 55 patients. The incidence of pulse strength discrepancies between feet was 20%, loss of pulse was 3.4% (6.7% in neonates, 1.4% in older infants) when extracorporeal membrane oxygenation patients were excluded, and resolution of pulse discrepancy or loss was 100%. Duration of catheterization and use of a catheter larger than 2.5 Fr were significant predictors of loss of pulse. Loss of pedal pulse distal to small-bore monitoring femoral artery catheters in neonates and infants is directly related to the duration of catheterization and is less frequent when 2.5-F, 5-cm polyethylene catheters are used instead of larger catheters.


Purification of peroxisomal fraction from rat brain.
A purification procedure to obtain peroxisomes (microperoxisomes) from the brain of suckling rats is reported. A P2 fraction, (crude light mitochondria) frozen and thawed seven times, was subfractionated yielding a P4 fraction, 4-fold enriched in catalase activity with respect to the cytoplasmic extract S1. The P4 fraction was used for further purification of peroxisomes by isopicnic centrifugation on Nycodenz gradient (1.10-1.20 g/ml). When the cerebellum was not included in the starting material, the equilibrium density of peroxisomes was 1.152-1.162 g/ml. In this case the overall yield of catalase in the most enriched fraction was 7% and its relative specific activity more than 50. When the cerebellum was included in the total homogenate, the equilibrium density shifted towards higher values (1.177 g/ml) and in this case the catalase relative specific activity in the peroxisomal enriched fraction was extremely high (> 100). The biochemical results, together with the electron microscope examination of the purified fractions, demonstrate that our procedure allows the best purification of brain peroxisomes so far obtained. The different equilibrium densities of peroxisomes observed in the two sets of experiments are interpreted in terms of size heterogeneity of these organelles in different brain portions and cell types.


68Ga-labelled-exendin-4: New GLP1R targeting agents for imaging pancreatic β-cell and insulinoma.
Glucagon-like peptide-1 receptor (GLP1R) specifically expressed on the surface of pancreatic β-cells and insulinoma, is a potential biomarker for imaging β-cell mass (BCM). In this study, two new 68Ga-labelled GLP1R targeting agents were prepared and their biological properties for imaging BCM and insulinoma were evaluated. [68Ga]Ga-HBED-CC-MAL-Cys39-exendin-4 ([68Ga]Ga-4) and its dimer ([68Ga]Ga-5) were synthesized from corresponding precursors. Cell uptake studies were evaluated in INS-1 cells. Biodistribution and microPET studies were performed in male normal Sprague-Dawley rats, diabetic rats and insulinoma xenograft NOD/SCID mice. [68Ga]Ga-4 and [68Ga]Ga-5 were efficiently radiolabelled by a simple one-step reaction without purification leading to high radiochemical yields and radiochemical purities (both >95%, decay corrected, n = 6, molar activity 15 GBq/μmol). They both showed excellent stability (~95%) in phosphate-buffered saline, pH 7.4, and in rat serum (~90%) for 2 h. Biodistribution studies and small animal PET/CT imaging showed that [68Ga]Ga-4 displayed specific uptake in rat pancreas and mouse insulinoma, and a reduced uptake in the pancreas of diabetic rat was observed (~62% reduction). Notably, it exhibited a rapid time-to-peak pancreatic uptake (0.96 ± 0.19%ID/g in 15 min) and fast clearance from the kidney (42% clearance in 30 min). Results suggested a favorable in vivo kinetics for human imaging studies. [68Ga]Ga-4 targeting GLP1R of pancreatic β-cells may be a potentially useful PET agent and a suitable candidate for further structural modification studies. This agent has demonstrated several advantages, rapid time-to-peak pancreatic uptake and faster clearance from the kidney, factors may enhance diagnosis of diabetes and insulinoma.


[Study on the chemical constituents of Phlomis younghusbandii].
To study the chemical constituents of Phlomis younghusbandii. Column chromatography was used in the isolation procedure. The structures of isolated compounds were elucidated by spectral data Eight compounds were isolated and their structures were identified as barlerin (1), sesamoside (2), phloyoside II (3), salidroside (4), shanzhiside methyl ester (5), acteoside (6), luteolin-7-O-glucoside (7), daucosterol (8). Compound 4 is firstly isolated from Phlomis genus. Compound 4 and 6 are obtained from this plant for the first time.


Sympathectomy-induced blood pressure reduction in adult normotensive and hypertensive rats is counteracted by enhanced cardiovascular sensitivity to vasoconstrictors.
The effect of chemical sympathectomy on cardiovascular parameters and the compensatory role of adrenal hormones, the renin-angiotensin system, and cardiovascular sensitivity to vasoconstrictors were studied in spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto (WKY) rats. Sympathectomy was induced in 20-week-old rats by daily intraperitoneal guanethidine administration (30 mg/kg b.w.) for 2 weeks. Basal blood pressure (BP), heart rate (HR), and restraint stress-induced cardiovascular changes were measured by radiotelemetry. The BP response to catecholamines was determined in rats with implanted catheters. Sympathectomy decreased BP only transiently, and after 14-day guanethidine treatment, BP returned to basal values in both strains. Sympathectomy permanently lowered HR, improved baroreflex sensitivity, and decreased the low-frequency domain of systolic blood pressure variability (a marker of vascular sympathetic activity). Guanethidine also attenuated the BP and HR responses to restraint stress. On the other hand, the BP response to catecholamines was augmented in sympathectomized rats, and this was not due to the de novo synthesis of vascular adrenergic receptors. Sympathectomy caused adrenal enlargement, enhanced the expression of adrenal catecholamine biosynthetic enzymes, and elevated plasma adrenaline levels in both strains, especially in WKY rats. Guanethidine also increased the plasma levels of aldosterone and corticosterone in WKY rats only. In conclusion, sympathectomy produced a transient decrease in BP, a chronic decrease in HR and improvement in baroreflex sensitivity. The effect of sympathectomy on BP was counteracted by increased vascular sensitivity to catecholamines in WKY rats and SHRs and/or by the enhanced secretion of adrenal hormones, which was more pronounced in WKY rats.


Culturally congruent mentorship can reduce disruptive behavior among elementary school students: results from a pilot study.
Our study objective was to examine the feasibility of implementing a culturally congruent mentorship pilot program, Youth-First (YF), that targets behavior modification among elementary school-aged children with disruptive behavior and a history of school suspension. We hypothesize that it is feasible to implement the YF program to reduce disruptive behaviors and recidivism of level III/IV infractions in school settings among at-risk African American students. We assessed program feasibility based on the success of program acceptance by parents/guardians, study enrollment, and intervention compliance by students. A pre/posttest study design was used to examine whether the YF program reduced recidivism of disruptive behavior among enrolled at-risk African American elementary school children between September 2016 and January 2017. Generalized linear mixed models examined whether student behavioral scores improved over time and varied by program mentor. A McNemar test examined the reduction in cumulative incidence of level III/IV infractions pre-post YF program intervention. Intervention acceptance, enrollment, and compliance were 100% (95% confidence interval [CI] 86 to 100%), 100% (95% CI 86 to 100%), and 67% (95% CI 45 to 84%), respectively (N = 24). Overall, student behavioral scores improved and plateaued over time (Time2 effect: b = - 0.01, 95% CI - 0.02, < 0.01); a two-week period was associated with a seven-point improvement (effect size: Cohen's d = 0.47, 95% CI 0.03, 0.94) in behavioral scores. Behavioral score improvements were class-specific, based on respectfulness behavior (b = 0.11, 95% CI < 0.01, 0.26). No recidivism of level III/IV infractions was reported during and post YF intervention. The integration of culturally congruent mentorship in elementary school-settings is feasible and can reduce risk of disruptive behaviors among at-risk African American students. Future studies should use randomized clinical trials to determine the effectiveness of culturally congruent mentorship interventions (void of potential selection and confounding biases) in reducing disruptive behavior, level III/IV infractions, and school suspensions among at-risk children.


The combination of metformin with morin alleviates hepatic steatosis via modulating hepatic lipid metabolism, hepatic inflammation, brown adipose tissue thermogenesis, and white adipose tissue browning in high-fat diet-fed mice.
The valuable effects of metformin (MET) and morin (MOR) in the improvement of NAFLD have been proposed, nevertheless, their combination impacts were not investigated so far. We determined the therapeutic effects of combined MET and MOR treatment in high-fat diet (HFD)-induced Non-alcoholic fatty liver disease (NAFLD) mice. C57BL/6 mice were fed on an HFD for 15 weeks. Animals were allotted into various groups and supplemented with MET (230 mg/kg), MOR (100 mg/kg), and MET + MOR (230 mg/kg + 100 mg/kg). MET in combination with MOR reduced body and liver weight in HFD-fed mice. A significant decrease in fasting blood glucose and improvement in glucose tolerance was observed in HFD mice treated with MET + MOR. Supplementation with MET + MOR led to a decline in hepatic triglyceride levels and this impact was associated with diminished expression of fatty-acid synthase (FAS) and elevated expression of carnitine palmitoyl transferase 1 (CPT1) and phospho-Acetyl-CoA Carboxylase (p-ACC). Moreover, MET combined with MOR alleviates hepatic inflammation through the polarization of macrophages to the M2 phenotype, decreasing the infiltration of macrophages and lowering the protein level of NF-kB. MET and MOR in combination reduce the size and weight of epididymal white adipose tissue (eWAT), and subcutaneous WAT (sWAT), whereas improves cold tolerance, BAT activity, and mitochondrial biogenesis. Combination therapy results in stimulating brown-like adipocyte (beige) formation in the sWAT of HFD mice. These results suggest that the combination of MET and MOR has a protective effect on hepatic steatosis, which may use as a candidate therapeutic for the improvement of NAFLD.


Interphotoreceptor retinoid-binding protein: role in delivery of retinol to the pigment epithelium.
The ability of interphotoreceptor retinoid-binding protein (IRBP) to facilitate the incorporation of retinol into retinyl esters by the retinal pigment epithelium (RPE) was examined in toad (Bufo marinus) eyecup preparations devoid of neural retina (RPE-eyecup). Solutions containing purified bovine IRBP and all-trans[3H]retinol were introduced into the vitreal cavity of the RPE-eyecup. After incubation at 22 degrees C, [3H]retinyl ester was extracted from the RPE cells and isolated by high performance liquid chromatography. All-trans[3H]retinyl ester formed in the RPE increased with time of incubation (up to 2 hr) and with concentration of IRBP (up to 10 microM). The increase with IRBP concentration accompanied, and presumably resulted from, an increased transfer of [3H]retinol to the RPE-eyecup. With higher concentration of IRBP (20-30 microM), both the amount of [3H]retinyl ester formed (relative to the peak value at 10 microM IRBP) and the overall molar content of endogenous retinyl ester were reduced. On the other hand, bovine serum albumin at relatively high concentration (90 microM) was less effective than 3 microM IRBP in supporting the formation of [3H]retinyl ester, and it did not reduce the level of native retinyl ester in the RPE. Using 3 microM IRBP, levels of [3H]retinyl ester formed were comparable to or exceeded those obtained with phosphatidyl choline (0.9 mg ml-1) or serum retinol-binding protein (3 microM). The data are consistent with the hypothesized role of IRBP as a carrier of retinol between the retina and RPE in the operation of the visual cycle.


Adherence to oxidative balance scores and lower odds of non-alcoholic fatty liver disease: a case-control study.
Evidence has also shown that oxidative stress and systemic inflammation, or in other words, disruption of the oxidant and antioxidant balance, can play an important role in the initiation or progression of NAFLD. The purpose of this study was to investigate the associations between the oxidative balance scores (OBS) and the risk of NAFLD. 552 healthy and 340 patients adult over the age of 18 with NAFLD participated in this case-control research. A validated 168-item quantitative food frequency questionnaire (FFQ) and indicators of physical activity, obesity, and smoking status were used to assess OBS score. The connection between OBS and NAFLD was discovered using binary logistic regression. The mean (± SD) age and (body mass index) BMI of the study population was 40.22 ± 9.79 years and 29.06 ± 3.92 kg/m2, respectively. The mean ± SD of OBS was 41.48 ± 5.23. After adjustment for potential confounders, higher scores of adherence to the OBS conferred a protection for the presence of NAFLD (odds ratio [OR]: 0.29; 95% confidence interval [CI]: 0.15-0.49; P for trend < 0.001). The findings of the present study indicate an approximately 80% reduction in the odds of developing NAFLD with higher OBS adherence in the overall population. However, prospective studies are needed to further investigate this association.


European standard clinical practice - Key issues for the medical care of individuals with familial leukemia.
Although hematologic malignancies (HM) are no longer considered exclusively sporadic, additional awareness of familial cases has yet to be created. Individuals carrying a (likely) pathogenic germline variant (e.g., in ETV6, GATA2, SAMD9, SAMD9L, or RUNX1) are at an increased risk for developing HM. Given the clinical and psychological impact associated with the diagnosis of a genetic predisposition to HM, it is of utmost importance to provide high-quality, standardized patient care. To address these issues and harmonize care across Europe, the Familial Leukemia Subnetwork within the ERN PaedCan has been assigned to draft an European Standard Clinical Practice (ESCP) document reflecting current best practices for pediatric patients and (healthy) relatives with (suspected) familial leukemia. The group was supported by members of the German network for rare diseases MyPred, of the Host Genome Working Group of SIOPE, and of the COST action LEGEND. The ESCP on familial leukemia is proposed by an interdisciplinary team of experts including hematologists, oncologists, and human geneticists. It is intended to provide general recommendations in areas where disease-specific recommendations do not yet exist. Here, we describe key issues for the medical care of familial leukemia that shall pave the way for a future consensus guideline: (i) identification of individuals with or suggestive of familial leukemia, (ii) genetic analysis and variant interpretation, (iii) genetic counseling and patient education, and (iv) surveillance and (psychological) support. To address the question on how to proceed with individuals suggestive of or at risk of familial leukemia, we developed an algorithm covering four different, partially linked clinical scenarios, and additionally a decision tree to guide clinicians in their considerations regarding familial leukemia in minors with HM. Our recommendations cover, not only patients but also relatives that both should have access to adequate medical care. We illustrate the importance of natural history studies and the need for respective registries for future evidence-based recommendations that shall be updated as new evidence-based standards are established.


Scattered accumulation hotspots of macro-litter on the seafloor: Insights for mitigation actions.
Marine litter is an ever-increasing problem that demands immediate reduction plans and mitigation actions that should act synergically to efficiently meet ambitious goals. Since the seafloor has been recognized as the major sink for marine debris, the study of litter accumulation dynamics represents a fundamental tool to evaluate future removal actions. We analysed a 7 years (2013-2019) standardized data series collected along Sardinian fishing grounds through MEDiterranean International Trawl Survey, for which estimates of density and weight of seafloor macro-litter were calculated over 707 hauls. Results show the absence of any temporal trend in seafloor macro-litter density and weight, but rather indicate a spatial and bathymetric segregation of different litter categories. Our data showed how different sources and physical features of macro-litter items (i.e., plastic and fishing gears, rubber, glass, metal and textile) led to spatially segregated accumulation hotspots. These hotspots often occurred at shallower depths and closer to coastlines, representing spots where future litter removal action could be prioritized. We also point out here how the identification of seafloor macro-litter hotspots using aggregated data that include plastic items could indeed hide the identification of hotspots of other less abundant but yet detrimental macro-litter categories accumulated in the marine environment.


Habituation to a Deterrent Plant Alkaloid Develops Faster in the Specialist Herbivore Helicoverpa assulta Than in Its Generalist Congener Helicoverpa armigera and Coincides with Taste Neuron Desensitisation.
The two closely related moth species, Helicoverpa armigera and H. assulta differ strongly in their degree of host-plant specialism. In dual-choice leaf disk assays, caterpillars of the two species that had been reared on standard artificial diet were strongly deterred by the plant-derived alkaloid strychnine. However, caterpillars of both species reared on artificial diet containing strychnine from neonate to the 5th instar were insensitive to this compound. Fifth instar caterpillars of H. assulta and 4th or 5th instars of H. armigera not exposed to strychnine before were subjected to strychnine-containing diet for 24 h, 36 h, 48 h, or 72 h. Whereas H. assulta displayed habituation to strychnine after 48 h, it took until 72 h for H. armigera to become habituated. Electrophysiological tests revealed that a deterrent-sensitive neuron in the medial sensillum styloconicum of both species displayed significantly reduced sensitivity to strychnine that correlated with the onset of habituation. We conclude that the specialist H. assulta habituated faster to strychnine than the generalist H. armigera and hypothesis that desensitization of deterrent-sensitive neurons contributed to habituation.


Thromboxane receptor antagonism and synthase inhibition in cerebral ischemia.
Thromboxane A2 (TXA2) is a proaggregatory vasoconstrictor that may suppress regional cerebral blood flow (rCBF) during postischemic hypoperfusion. This study was undertaken to determine if rCBF could be elevated by postischemic treatment with a TXA2 receptor antagonist, SQ29,548, given alone or in combination with 1-benzylimidazole (1-BI), a thromboxane synthase inhibitor. Wistar rats were subjected to 30 min of reversible forebrain ischemia and treated with SQ29,548 or an SQ29,548/1-BI combination during 60 min of reperfusion. Cerebral TXB2, the stable metabolite of TXA2, was 1.33 +/- 0.91 ng mg brain protein-1 in animals treated with SQ29,548 and exposed to ischemia, compared to 1.15 +/- 0.32 in ischemic controls (p = NS). Administration of SQ29,548/20 mg kg-1 1-BI reduced cerebral TXB2 to 0.20 +/- 0.25 (p < or = 0.01). Regional CBF was depressed significantly in ischemic controls compared to sham-ischemic animals (p < or = 0.01 in all regions except for p < or = 0.05 in diencephalon) and was not altered by treatment with SQ29,548. Rats given the SQ29,548/1-BI combination showed an overall increase in rCBF that did not reach statistical significance when compared to ischemic controls. However, rCBF in hippocampus and diencephalon of animals given the drug combination was significantly greater than in rats treated with SQ29,548 alone (p < or = 0.05).


Ionomycin ameliorates hypophosphatasia via rescuing alkaline phosphatase deficiency-mediated L-type Ca2+ channel internalization in mesenchymal stem cells.
The loss-of-function mutations in the ALPL result in hypophosphatasia (HPP), an inborn metabolic disorder that causes skeletal mineralization defects. In adults, the main clinical features are early loss of primary or secondary teeth, osteoporosis, bone pain, chondrocalcinosis, and fractures. However, guidelines for the treatment of adults with HPP are not available. Here, we show that ALPL deficiency caused a reduction in intracellular Ca2+ influx, resulting in an osteoporotic phenotype due to downregulated osteogenic differentiation and upregulated adipogenic differentiation in both human and mouse bone marrow mesenchymal stem cells (BMSCs). Increasing the intracellular level of calcium in BMSCs by ionomycin treatment rescued the osteoporotic phenotype in alpl+/- mice and BMSC-specific (Prrx1-alpl-/-) conditional alpl knockout mice. Mechanistically, ALPL was found to be required for the maintenance of intracellular Ca2+ influx, which it achieves by regulating L-type Ca2+ channel trafficking via binding to the α2δ subunits to regulate the internalization of the L-type Ca2+ channel. Decreased Ca2+ flux inactivates the Akt/GSK3β/β-catenin signaling pathway, which regulates lineage differentiation of BMSCs. This study identifies a previously unknown role of the ectoenzyme ALPL in the maintenance of calcium channel trafficking to regulate stem cell lineage differentiation and bone homeostasis. Accelerating Ca2+ flux through L-type Ca2+ channels by ionomycin treatment may be a promising therapeutic approach for adult patients with HPP.


Advancing Family Presence Policies and Practices in the Canadian Health and Care Context: COVID-19 and Beyond.
With the onset of the COVID-19 pandemic, restrictive visitor policies have curtailed the ability of family caregivers to be present to partner in the care of loved ones. Building on the success of the "Better Together" campaign, Healthcare Excellence Canada - the newly amalgamated organization of the Canadian Foundation for Healthcare Improvement and the Canadian Patient Safety Institute - has co-developed policy guidance and "Essential Together" programming that recognizes the significant role of essential care partners. This work aims to support the safe reintegration of essential care partners into health and care organizations across Canada during the pandemic and beyond.


[Phage-mediated conjugative transfer of plasmids in Staphylococcus aureus].
It was shown possible to transfer nonconjugative plasmids during joint cultivation of the donor and recipient cells by transduction and phage-mediated conjugation. In the latter case it was necessary that the phage in the medium was free and the prophage was present in the recipient cells. Differences in the regularities of the transfer of the nonconjugative plasmids mobilized by the conjugative plasmid or phage were observed.


Permanent Unfitness for Work and Musculoskeletal Diseases: A Multicentric Cross-sectional Study of 2788 Unfit Employees.
To describe the various musculoskeletal diseases resulting in work-related permanent unfitness for work. Methods: In 2017, a multicenter cross-sectional employee survey in the South of France was conducted by 323 occupational physicians. Results: Of the 359,966 employees monitored, 2788 had permanent unfitness for work. This number included 1078 for musculoskeletal diseases, which was more frequent in women (odds ratio, 1.42; confidence interval, 1.20 to 1.68) and showed an increase in women older than 55 years (odds ratio, 6.42; confidence interval, 4.4 to 6.60). Back disorder was the most common work-related disorder reported (n = 488 [45.4%]), with soft tissue conditions being (n = 237 [22.0%]). Arthropathy (n = 157 [14.6%]) had occupational origins in 282 (44.3%), 169 (26.6%), and 55 (8.6%) cases, respectively. Conclusions: Musculoskeletal disability prevention should be encouraged to avoid resultant employment attrition.


Evaluation of a concentric biopsychosocial model of well-being in persons with spinal cord injuries.
The objective of this study was to evaluate a concentric biopsychosocial model of well-being in individuals with spinal cord injuries (SCI). Adults (N = 235) with SCI participated in this study. A cross-sectional design with hierarchical regression and Andrew Hayes' (2013) PROCESS mediation analysis procedure was used to evaluate the model. Each step of the hierarchical regression on life satisfaction, in which biological variables were entered first, social variables were entered second, and psychological variables were entered third, was significant. Examining the standardized partial coefficients, pain, interpersonal self-efficacy, social support, hope-agency, and self-esteem were all significantly associated with life satisfaction, controlling for variables in each outward ring of the concentric model. Four serial mediational analyses were also conducted in which the social and psychological variables significantly partially mediated the relationship between pain and life satisfaction. The results provide support for a concentric biopsychosocial model of well-being in persons with SCI. Rehabilitation interventions should focus on augmenting biopsychosocial factors to allow for maximum improvement in well-being outcomes in individuals with SCI. (PsycINFO Database Record