Lactobacillus rhamnosus GR-1 stimulates colony-stimulating factor 3 (granulocyte) (CSF3) output in placental trophoblast cells in a fetal sex-dependent manner.
Bacterial vaginosis is associated with a 1.4-fold increased risk of preterm birth. We have shown previously that Lactobacillus rhamnosus GR-1 supernatant up-regulates interleukin 10 and down-regulates tumor necrosis factor-alpha output in lipopolysaccharide (LPS)-treated human primary placenta cultures in a fetal sex-dependent manner. We hypothesize that lactobacilli also exert their anti-inflammatory effect by up-regulation of colony-stimulating factor 3 (granulocyte) (CSF3), which is secreted from both immune and placental trophoblast cells, and that this activity is dependent on the sex of the fetus. Placental trophoblast cells were isolated from term elective cesarean section placentae using a Percoll gradient and separated from CD45(+) cells using magnetic purification. Cells were treated with LPS in the presence or absence of pretreatments with L. rhamnosus GR-1 supernatant or chemical inhibitors of the intracellular signaling pathways. Phosphorylations of mitogen-activated protein kinase 14 (MAPK14, previously known as p38) and signal transducer and activator of transcription (STAT) 3 were measured by Western blot analysis, and levels of CSF3 were determined by ELISA. CSF3 output was increased only in the placental trophoblast cells of female fetuses treated with LPS, GR-1 supernatant, and a combination of both treatments. The GR-1 supernatant up-regulated the phosphorylation of STAT3 and MAPK14. CSF3 output was inhibited by both Janus kinases (JAK) and MAPK14 inhibitors. None of the treatments was able to increase CSF3 output in either the pure trophoblast or the CD45(+) cell preparations alone. These results suggest an underlying mechanism for the sex difference in incidence of preterm birth and provide potential evidence for a therapeutic benefit of lactobacilli in reducing the risk of preterm labor.


Comparison of different cardiovascular risk tools used in HIV patient cohorts in sub-Saharan Africa; do we need to include laboratory tests?
Cardiovascular disease (CVD) is the leading cause of death globally, representing 31% of all global deaths. HIV and long term anti-retroviral therapy (ART) are risk factors for development of CVD in populations of people living with HIV (PLHIV). CVD risk assessment tools are currently being applied to SSA populations, but there are questions about accuracy as well as implementation challenges of these tools in lower resource setting populations. We aimed to assess the level of agreement between the various cardiovascular screening tools (Data collection on Adverse effects of anti-HIV Drugs (D:A:D), Framingham risk score, WHO risk score and The Atherosclerotic Cardiovascular Disease Score) when applied to an HIV ART experienced population in Sub-Saharan Africa. This study was undertaken in an Anti-Retroviral Long Term (ALT) Cohort of 1000 PLHIV in care who have been on ART for at least 10 years in urban Uganda. A systematic review was undertaken to find the most frequently used screening tools from SSA PLHIV populations; these were applied to the ALT cohort. Levels of agreement between the resulting scores (those including lipids and non-lipids based, as well as HIV-specific and non-HIV specific) as applied to our cohort were compared. Prevalence Bias Adjusted Kappa was used to evaluate agreement between tools. Overall, PLHIV in ALT cohort had a median score of 1.1-1.4% risk of a CVD event over 5 years and 1.7-2.5% risk of a CVD event over 10 years. There was no statistical difference in the risk scores obtained for this population when comparing the different tools, including comparisons of those with lipids and non-lipids, and HIV specific vs non-HIV specific. The various tools yielded similar results, but those not including lipids are more feasible to apply in our setting. Long-term cohorts of PLHIV in SSA should in future provide longitudinal data to evaluate existing CVD risk prediction tools for these populations. Inclusion of HIV and ART history factors to existing scoring systems may improve accuracy without adding the expense and technical difficulty of lipid testing.


Synergistic fungistatic effects of lactoferrin in combination with antifungal drugs against clinical Candida isolates.
Because of the rising incidence of failures in the treatment of oropharyngeal candidosis in the case of severely immunosuppressed patients (mostly human immunodeficiency virus [HIV]-infected patients), there is need for the development of new, more effective agents and/or compounds that support the activity of the common antifungal agents. Since lactoferrin is one of the nonspecific host defense factors present in saliva that exhibit antifungal activity, we studied the antifungal effects of human, bovine, and iron-depleted lactoferrin in combination with fluconazole, amphotericin B, and 5-fluorocytosine in vitro against clinical isolates of Candida species. Distinct antifungal activities of lactoferrin were observed against clinical isolates of Candida. The MICs generally were determined to be in the range of 0.5 to 100 mg. ml(-1). Interestingly, in the combination experiments we observed pronounced cooperative activity against the growth of Candida by using lactoferrin and the three antifungals tested. Only in a limited concentration range was minor antagonism detected. The use of lactoferrin and fluconazole appeared to be the most successful combination. Significant reductions in the minimal effective concentrations of fluconazole were found when it was combined with a relatively low lactoferrin concentration (1 mg/ml). Such combinations still resulted in complete growth inhibition, while synergy of up to 50% against several Candida species was observed. It is concluded that the combined use of lactoferrin and antifungals against severe infections with Candida is an attractive therapeutic option. Since fluconazole-resistant Candida species have frequently been reported, especially in HIV-infected patients, the addition of lactoferrin to the existing fluconazole therapy could postpone the occurrence of species resistance against fluconazole. Clinical studies to further elucidate the potential utility of this combination therapy have been initiated.


A new evaluation framework for the assessment of wastewater heat recovery potential coupled with wastewater reuse.
The integration of wastewater heat recovery (WWHR) and wastewater reuse offers a numerous advantage, making its application possible in various sectors. Nevertheless, this concept faced challenges to the identification of appropriate location. Existing research lacks comprehensive evaluation methods that encompass a various factor for effective decision-making. This study introduces a new evaluation framework that involves different aspects, including thermal energy potential and spatial distribution analysis. The novelty of this research lies in its unique focus on the combination of WWHR and wastewater reuse. Moreover, it introduces a structured evaluation framework that considers multiple criteria and expert opinions, enhancing decision-making precision. Multi-criteria decision analysis (MCDA) was applied to select assessment criteria, which were categorized into three aspects: water-energy supplier, water-energy consumers, and water-energy station. The relative importance of criteria was determined using the analytical hierarchical process (AHP). The results of the AHP highlight significance of factors: treated wastewater flow rate; treated wastewater temperature; water-energy supply distance, and type of water-energy consumer. These factors were assigned weight values of 0.297, 0.186, 0.123, and 0.096, respectively. It is emphasizing their influence in the decision-making process that potential locations depend on the water-energy supplier and water-energy consumer as supply and demand sources.


Correlation between serotonin synthesis and 5-HT1A receptor binding in the living human brain: a combined alpha-[11C]MT and [18F]MPPF positron emission tomography study.
Serotonin (5-HT) is one of the major neurotransmitters and has been implicated in a wide variety of cerebral functions. Several lines of evidence indicate that 5HT(1A) receptors exert a negative feedback in the synthesis and release of serotonin. While most of what is known about serotonin comes from studies in animals, much less empirical evidence exists about the serotonergic system in the living human brain. This study aims to assess the correlation between serotonin synthesis and 5-HT(1A) receptor binding using positron emission tomography (PET) in humans. Six healthy male volunteers underwent 2 PET scans in the same day: one measuring alpha-[(11)C]MT K [ml/g/min] trapping constant (a measure of serotonin synthesis) and one measuring 5-HT(1A) receptor binding potential BP(ND) with [(18)F]MPPF. Volumes of interest (VOIs) selected a priori included: anterior cingulate cortex (ACC), anterior insula, hippocampus, amygdala, thalamus, hypothalamus and midbrain raphe nuclei. Correlation analyses were conducted voxel-by-voxel and with manually traced VOIs. A significant negative correlation between serotonin synthesis and 5-HT(1A) binding potential was found bilaterally in hippocampus and anterior insula and in the left ACC. The combination of [(18)F]MPPF and alpha-[(11)C]MT PET offers a means to investigate key determinants of 5-HT neurotransmission under physiological and psychopathological conditions in the human brain in vivo.


[Transplantation of somatic nuclei into activated teleost eggs (the example of the loach, Misgurnus fossilis L.)].
Some peculiarities of the technique of nuclear transplantation for teleostean fishes are described, the loach (Misgurnus fossilis) taken as an example. The cells of the late high blastula were used as donors and the activated non-enucleated and enucleated (by X-rays, 20 kR) eggs of the loach as recipients. Following the nuclear transplantation in the activated non-enucleated eggs, 33 (out of 128) normal blastulae were obtained, one of which developed until hatching. Following the nuclear transplantation in the activated enucleated eggs, 34 (out of 251) blastulae were obtained, 6 embryos hatched and 2 larvae attained the stage of active feeding.


Cluster-phase reactions: gas-phase phosphorylation of peptides and model compounds with triphosphate anions.
Molecular clusters provide a unique environment in which chemical reactions between cluster components can occur. In the present study, electrospray ionization is used to examine the behavior of anionic clusters of triphosphate with choline, acetylcholine, and betaine, and the behaviors of cationic clusters of triphosphate with the peptides bradykinin (RPPGFSPFR) and ARRPEGRTWAQPGY. Phosphorylation of a hydroxyl group, when one is present, is shown to be a facile process when the cluster is subjected to collisional activation. Of particular interest is the selective phosphorylation of the hydroxyl substituent in serine and threonine residues of peptides. Less conclusive results are obtained with three peptides containing tyrosine, but the data obtained are consistent with phosphorylation on tyrosine residues. In the absence of residues with hydroxyl substituents, the C-terminus of a peptide is observed to be phosphorylated. The unique chemical reactions reported in this study represent the first examples of gas-phase phosphorylation of alcohols and are also interesting in that they occur at a site remote from charged functional groups in the same molecule. This facile process may have interesting implications for the synthesis of key molecules at the threshold of life.


'This is mine, this is for me': preexposure prophylaxis as a source of resilience among women in Eswatini.
To understand the relationship between HIV preexposure prophylaxis (PrEP) and resilience among Emaswati women. A qualitative study using semistructured in-depth interviews. We interviewed 24 women who recently initiated PrEP at one of six public-sector primary-care clinics participating in a government demonstration project for PrEP for the general population, as well as 30 PrEP stakeholders from HIV policy, implementation and donor sectors. PrEP clients and stakeholders described an environment marked by high HIV risk. In this context, clients felt PrEP enhanced their resilience through feelings of protection, control over HIV acquisition, choice in relation to when to take PrEP, sexual pleasure and relief from the fear and consequences of HIV infection. Those in serodiscordant relationships described improved partner communication and a shared sense of responsibility for HIV prevention. Both PrEP clients and stakeholders asserted that PrEP marketing and communication should be Emaswati-led and community-owned. PrEP helps women in Eswatini envision longer, healthier and more fulfilled lives. PrEP enhances resilience through several pathways, including self-efficacy, choice, control over HIV infection and relief from the fear of HIV infection. Social marketing and information campaigns should not only focus on PrEP as an HIV-prevention method but also emphasize how PrEP has expanded choice, enhanced control and facilitated sexual enjoyment among PrEP clients.


Cytological Findings of Ascitic Fluid with a Malignant Ovarian Steroid Cell Tumor: A Case Report and Literature Review.
Ovarian steroid cell tumors (SCTs) are rare and usually benign, although 25-43% are reportedly malignant. The cytologic findings of these rare ovarian tumors have almost never been reported. We report a rare case of a malignant ovarian SCT with peritoneal dissemination and malignant ascites in a 40-year-old woman. Her tumor was classified as stage IIB (pT2bNoM0) according to the FIGO (International Federation of Gynecology and Obstetrics) classification system, and she was treated with adjuvant chemotherapy following staging laparotomy. Cytology of the ascitic fluid revealed large, polygonal-to-round cells and multinucleated cells with atypia, appearing in clusters with slight overlapping or as isolated tumor cells. Numerous tumor cells had small central round or eccentric nuclei with conspicuous nucleoli, and a moderate-to-abundant amount of cytoplasm, varying from granular and eosinophilic to pale and multivacuolated (foamy), with cannibalism formations. The nuclear chromatin was fine and granular, with irregular distribution and nuclear-membrane thickening. These may be the first reported cytology results for ascites with a malignant SCT. Our patient's cytological ascitic findings, rather than the histopathologic features of the original and disseminated tumors, represent the malignant features of the tumor.


Diffuse intrapulmonary malignant mesothelioma masquerading as interstitial lung disease: a distinctive variant of mesothelioma.
Malignant mesothelioma typically encases lungs as a thick rind, while relatively sparing lung parenchyma. We describe an unusual presentation of mesothelioma characterized by diffuse intrapulmonary growth, with absent or inconspicuous pleural involvement, clinically simulating interstitial lung disease (ILD). We identified 5 patients (median age 56 y, all men) with diffuse intrapulmonary malignant mesothelioma in our pathology consultation practice from 2009 to 2012. Clinical history, imaging, and pathology materials were reviewed. Symptoms included chronic dyspnea (4 cases), cough (3), and acute dyspnea with bilateral pneumothorax (1). Chest imaging showed irregular opacities (5), reticulation (4), pleural effusions (2), and subpleural nodular densities (1), without radiologic evidence of pleural disease or masses. A clinicoradiologic diagnosis of ILD was made in all cases, and wedge biopsies were performed. Histologic evaluation revealed a neoplastic proliferation of bland epithelioid or spindled cells, showing various growth patterns simulating silicotic nodules, desquamative interstitial pneumonia, organizing pneumonia, and Langerhans cell histiocytosis. Some areas mimicked adenocarcinoma, with lepidic, acinar, micropapillary, and solid patterns. Initial diagnoses by referring pathologists included reactive changes (1), hypersensitivity pneumonitis versus drug reaction (1), desquamative interstitial pneumonia versus neoplasm (1), and mesothelioma (2). Microscopic pleural involvement was identified in 4 cases. Immunohistochemistry confirmed the characteristic immunophenotype of mesothelioma in all cases. Median survival of 3 patients treated with chemotherapy was 28 months. Two patients received no therapy and survived 3 and 4 weeks, respectively. "Diffuse intrapulmonary malignant mesothelioma" is a rare variant with a distinctive presentation that clinically mimics ILD. Recognition is essential to avoid misdiagnosis.


Treatment of Spinal Infections.
The treatment of spinal infections is not well defined, and a cursory review of the literature can lead to conflicting treatment strategies. To add to the complexity, infections can include primary infection of the spine, infection secondary to another primary source, and postoperative infections including epidural abscesses, discitis, osteomyelitis, paraspinal soft-tissue infections, or any combination. Furthermore, differing opinions often exist within the medical and surgical communities regarding the outcomes and effectiveness of varying treatment strategies. Given the paucity of defined treatment protocols and long-term follow-up, it is important to develop multidisciplinary treatment teams and treatment strategies. This, along with defined protocols for the treatment of varying infections, can provide the data needed for improved treatment of spinal infections.


Alterations in peripheral purinergic and muscarinic signaling of rat bladder after long-term fructose-induced metabolic syndrome.
We explored the pathophysiologic mechanisms of long-term fructose-induced lower urinary tract symptoms (LUTS) in rats. Male Wistar rats were fed with fructose for 3 or 6 months. Biochemical and transcystometric parameters were compared between fructose-fed and age-matched normal-diet rats. Pelvic nerve and external urethral sphincter-electromyogram activity recordings were performed to investigate fructose effects on neural control of bladders. Mitochondrial structure, ATP and acetylcholine content and purinergic and muscarinic cholinergic receptors were examined. Cytosolic cytochrome C staining by Western blot and immunocytochemistry for mitochondrial injury and PGP 9.5 stain for nerve density were also determined. The fructose-fed rats with higher plasma triglyceride, LDL and fasting glucose levels displayed LUTS with increased frequency and suppressed voiding contractile amplitude in phase 1 and phase 2 duration versus normal-diet control. Fructose feeding altered the firing types in pelvic afferent and efferent nerves and external urethral sphincter-electromyogram activity. Increased mast cell number, disrupted and swollen mitochondria, increased cytosolic cytochrome C stain and expression and decreased nerve density in bladder smooth muscle layers appeared in the fructose-fed rats. Fructose feeding also significantly reduced ATP and acetylcholine content and enhanced protein expression of postsynaptic P(2)X(1), P(2)X(2) and P(2)X(3) purinergic receptors and M(2) and M(3) muscarinic cholinergic receptors expression in the smooth muscles of urinary bladder. Long-term fructose feeding induced neuropathy and myopathy in the urinary bladders. Impaired mitochondrial integrity, reduced nerve density, ATP and acetylcholine content and upregulation of purinergic and muscarinic cholinergic receptors expression may contribute to the bladder dysfunction of fructose-fed animals.


Mechanisms of mitochondrial holocytochrome c synthase and the key roles played by cysteines and histidine of the heme attachment site, Cys-XX-Cys-His.
Mitochondrial cytochrome c assembly requires the covalent attachment of heme by thioether bonds between heme vinyl groups and a conserved CXXCH motif of cytochrome c/c1. The enzyme holocytochrome c synthase (HCCS) binds heme and apocytochrome c substrate to catalyze this attachment, subsequently releasing holocytochrome c for proper folding to its native structure. We address mechanisms of assembly using a functional Escherichia coli recombinant system expressing human HCCS. Human cytochrome c variants with individual cysteine, histidine, double cysteine, and triple cysteine/histidine substitutions (of CXXCH) were co-purified with HCCS. Single and double mutants form a complex with HCCS but not the triple mutant. Resonance Raman and UV-visible spectroscopy support the proposal that heme puckering induced by both thioether bonds facilitate release of holocytochrome c from the complex. His-19 (of CXXCH) supplies the second axial ligand to heme in the complex, the first axial ligand was previously shown to be from HCCS residue His-154. Substitutions of His-19 in cytochrome c to seven other residues (Gly, Ala, Met, Arg, Lys, Cys, and Tyr) were used with various approaches to establish other roles played by His-19. Three roles for His-19 in HCCS-mediated assembly are suggested: (i) to provide the second axial ligand to the heme iron in preparation for covalent attachment; (ii) to spatially position the two cysteinyl sulfurs adjacent to the two heme vinyl groups for thioether formation; and (iii) to aid in release of the holocytochrome c from the HCCS active site. Only H19M is able to carry out these three roles, albeit at lower efficiencies than the natural His-19.


Evaluation of an Amino Acid Mix on the Secretion of Gastrointestinal Peptides, Glucometabolic Homeostasis, and Appetite in Obese Adolescents Administered with a Fixed-Dose or ad Libitum Meal.
Proteins have been demonstrated to reduce food intake in animals and humans via peripheral and central mechanisms. Supplementation of a dietetic regimen with single or mixed amino acids might represent an approach to improve the effectiveness of any body weight reduction program in obese subjects. The aim of the present study was to evaluate the effects of an amino acid mix (L-arginine + L-leucine + L-glutamine + L-tryptophan) on the secretion of some gastrointestinal peptides (i.e., ghrelin and glucagon-like peptide type 1, GLP-1), glucometabolic homeostasis (i.e., glucose, insulin, and glucagon), and appetite (hunger/satiety scored by visual analogue scale, VAS) in obese adolescents (n = 14; 10 females and 4 males; age: 16.6 ± 1.0 years; body mass index (BMI): 36.4 ± 4.6 kg/m²; fat-free mass (FFM): 54.9 ± 4.7%; fat mass (FM): 45.1 ± 4.4%) administered with a fixed-dose (lunch) or ad libitum (dinner) meal. Isocaloric maltodextrins were used as control treatment. During the lunch test, a significant increase in circulating levels of GLP-1, but not of ghrelin, was observed in the amino acid-treated group, which was congruent with significant changes in appetite, i.e., increase in satiety and decrease in hunger. A significant hyperglycemia was found in the maltodextrin-treated group during the prelunch period, without any significant changes in insulin and glucagon between the two groups. During the dinner test, there were no significant differences in appetite (hunger/satiety) and intake of calories. In conclusion, L-arginine, L-leucine, L-glutamine, and L-tryptophan, when administered to obese adolescents with a fixed-dose meal, are capable of evoking an anorexigenic response, which is, at least in part, mediated by an increase in GLP-1 released in circulation by L cells, which are capable of chemosensing specific amino acids present in the intestinal lumen. Further additional studies are requested to understand whether higher doses are necessary to inhibit ad libitum feeding.


Evaluation on in vitro biocompatibility of silk fibroin-based biomaterials with primarily cultured hippocampal neurons.
Silk fibroin-based biomaterials have recently found increasing applications in the tissue-engineering field including the generation of artificial nerve guides for peripheral nerve repair. The aim of this study was to investigate the suitability of silk fibroin as a candidate biomaterial for central nervous system (CNS) therapy. We found that substrates made up of silk fibroin fibers supported the survival and growth of the attached hippocampal neurons by using morphological observation. We also cultured the hippocampal neurons in silk fibroin extract for different times, and observed no significant difference occurring in their morphology, cell viability for these cultured hippocampal neurons as compared to those cultured in plain neuronal culture medium. Moreover, immunocytochemistry, RT-PCR, and Western blot analysis revealed that no significant difference was found in mRNA or protein levels of the growth-associated molecules, such as brain-derived neurotrophic factor, growth-associated protein-43, neurofilament, nerve growth factor, and nerve growth factor-receptor P75, between the hippocampal neurons cultured in the silk fibroin extract and in plain neuronal culture medium. Taken together, all the results demonstrate that silk fibroin has good biocompatibility with primarily cultured hippocampal neurons without any significant cytotoxic effects on their cell phenotype and functions, suggesting a potential possible use of silk fibroin for preparing the tissue-engineered nerve guides or drug delivery vehicles to treat CNS injuries or diseases.


Pathological characterization of keel bone fractures in laying hens does not support external trauma as the underlying cause.
Keel bone fractures in laying hens have been described with increasing prevalence from several countries over the last twenty years and are considered one of the greatest welfare problems to the layer industry. In Denmark we have observed fracture prevalence in the range of 53% to 100% in flocks from cage-free systems whereas flock prevalences in birds from enriched cages ranged between 50-98%. Previous research have speculated that the underlying reason for the development of keel bone fractures is trauma in relation to impact of the bird with furniture, other equipment etc. However, little evidence of this theory has been provided. Predisposing factors have also been suggested including genetics of the bird, lack of specific feedstuff components, high egg production, management factors and layer fatigue. This study has addressed the possible pathogenesis of these fractures by pathological characterization of fractures in birds from different production systems. More than 60 keel bones with fractures have been characterized histo-pathologically and by CT scan. This included an assessment of damage to muscles and soft tissues, the bone and the healing process including callus formation. This investigation has shown that high energy collisions cannot be responsible for the majority of fractures, located at the caudal tip of the keel bone, observed in laying birds as markers associated trauma were not observed in the majority of the cases just as few recognized healing processes were observed. These results suggest an alternative pathogenesis to trauma.


Stand-alone self-powered integrated microfluidic blood analysis system (SIMBAS).
We present a self-powered integrated microfluidic blood analysis system (SIMBAS) that does not require any external connections, tethers, or tubing to deliver and analyze a raw whole-blood sample. SIMBAS only requires the user to place a 5 μL droplet of whole-blood at the inlet port of the device, whereupon the stand-alone SIMBAS performs on-chip removal of red and white cells, without external valving or pumping mechanisms, followed by analyte detection in platelet-containing plasma. Five complete biotin-streptavidin sample-to-answer assays are performed in 10 min; the limit of detection is 1.5 pM. Red and white blood cells are removed by trapping them in an integral trench structure. Simulations and experimental data show 99.9% to 100% blood cell retention in the passive structure. Powered by pre-evacuation of its PDMS substrate, SIMBAS' guiding design principle is the integration of the minimal number of components without sacrificing effectiveness in performing rapid complete bioassays, a critical step towards point-of-care molecular diagnostics.


Importance of norepinephrine alpha 2-receptor activation for morphine-induced rat growth hormone secretion.
The normal pulsatile secretion of rat growth hormone (rGH) requires intact function in monoaminergic neurons. The importance of norepinephrine (NE) for the secretion is well documented, while the roles of dopamine (DA) and 5-hydroxytryptamine (5-HT) are still a matter of controversy. Morphine, as well as endogenous opioid peptides, are known to stimulate the secretion of GH. Whether the opiate-induced GH release is dependent on monoamines was investigated in the present study. Administration of morphine (10 mg/kg) resulted within 30 min in elevations of plasma rGH exceeding 40 ng/ml. This effect was almost completely antagonized by reserpine (10 mg/kg), given 5 h before morphine and partially antagonized by reserpine (2 mg/kg) administered 25 h before morphine. Administration of tetrabenazine (75 mg/kg) protects monoamine granules from irreversible destruction and counteracted the morphine antagonistic effects of reserpine (2 mg/dg) on rGH secretion indicating that the latter effect is due to blockade of monoaminergic neurotransmission. Pretreatment with either haloperidol (1 mg/kg), p-chlorophenylalanine (PCPA) (300 mg/kg x III) or phenoxybenzamine (10 mg/kg) did not reduce the effect of morphine on rGH release whereas yohimbine (3 mg/kg) effectively prevented it. In reserpine-pretreated animals administration of clonidine (0.5 mg/kg) potentiated the GH releasing effect of morphine. These results indicate that morphine-induced GH release is dependent on activation of postsynaptic NE receptors of the alpha 2-subtype.


Vagus nerve SARS-CoV-2 infection and inflammatory reflex dysfunction: Is there a causal relationship?
Autonomic dysfunction is a clinical hallmark of infection caused by SARS-CoV-2, but the underlying mechanisms are unknown. The vagus nerve inflammatory reflex is an important, well-characterized mechanism for the reflexive suppression of cytokine storm, and its experimental or clinical impairment facilitates the onset and progression of hyperinflammation. Recent pathological evidence from COVID-19 victims reveals viral infection and inflammation in the vagus nerve and associated nuclei in the medulla oblongata. Although it has been suggested that vagus nerve inflammation in these patients mediates dysregulated respiration, whether it also contributes to dysfunction of the vagus nerve inflammatory reflex has not been addressed. Because lethality and tissue injury in acute COVID-19 are characterized by cytokine storm, it is plausible to consider evidence that impairment of the inflammatory reflex may contribute to overproduction of cytokines and resultant hyperinflammatory pathogenesis. Accordingly, here the authors discuss the inflammatory reflex, the consequences of its dysfunction in COVID-19, and whether there are opportunities for therapeutic intervention.


Racial differences in the outcomes of patients with diastolic heart failure.
More than half of all patients with congestive heart failure have preserved left ventricular systolic function. This is particularly common in African American patients, yet there have been few studies examining the long-term natural history of this disorder in African-American and white patients. We studied 2740 white and 563 African American patients with class II to IV symptoms and preserved systolic function (ejection fraction >40) identified in the Duke Cardiovascular Databank from 1984 to 1996. Unadjusted and adjusted 5-year survival rate comparisons were performed with Kaplan-Meier and Cox proportional hazards models, respectively. The 5-year survival rates were 68% for African American patients and 70% for white patients (P =.55). However, after adjusting for known risk factors, African American patients had a significantly higher mortality risk than white patients (hazard ratio [HR], 1.34; 95% CI, 1.13-1.60). This racial difference in survival rate was most prominent in patients with a non-ischemic etiology (HR, 1.6; 95% CI, 1.2-2.0) as compared with patients with ischemic heart failure (HR, 1.1; 95% CI, 0.9-1.4). Among patients with heart failure and preserved left ventricular systolic function, African American patients have a worse long-term prognosis than white patients. These results are important because of the prevalence of this condition in African American patients and their potential heterogeneous response to many heart failure therapies.


Cefotaxime prevents microbial contamination and improves microspore embryogenesis in wheat and triticale.
Cefotaxime (100 mg/l) mitigate occasional gram negative bacterial contamination in wheat and triticale microspore culture and most importantly it increases cell growth and green plant production. Isolated microspore culture is a promising option to rapidly fix the product of meiotic recombination of F1 hybrids, in the process of varietal development. Clean culture and high embryogenesis rate are essential to commercial triticale and wheat microspore cultures. So, this study investigated (1) contaminants from isolated microspores cultures, (2) two antibiotics to control bacterial growth, and (3) the contribution of antibiotics to increased microspore-derived embryo-like structures (ELS), green and albino plants. Five species of bacteria were identified in contaminated cultures (Erwinia aphidicola, Pantoea agglomerans, Pseudomonas sp., Staphylococcus epidermis and Staphylococcus warneri) using fatty acid analysis and 16S ribosomal RNA sequences analysis, and yeast. Antibacterial susceptibility test using Cefotaxime and Vancomycin resulted in strong inhibition of 24 bacterial isolates, using Cefotaxime at 100 mg/l, but not Pseudomonas sp. Other antibiotic treatments inhibited bacterial growth at least partially. Microspore induction medium supplemented with the same antibiotics treatments resulted in successful microspore embryogenesis and green plant production. Antibiotic treatments were first tested in triticale and then validated in wheat cultivars AC Carberry and AC Andrew. Induction medium supplemented with Cefotaxime at 50 and 100 mg/l substantially increased the formation of ELS and green plants in triticale and wheat, respectively. Incidentally, it also affected the occurrence of albinism in all genotypes. Our results demonstrated dual purpose of Cefotaxime for isolated microspore culture, most importantly it increases cell growth and success of microspore cultures in triticale and wheat genotypes, but would also prevent accidental loss of cultures with most common bacterial contaminants.


Creation of a thermostable firefly luciferase with pH-insensitive luminescent color.
The thermal instability and pH-sensitive spectral property of firefly luciferase have hampered its use as a sensitive multicolor luminescent label or bioluminescent resonance energy transfer donor. With the intention of improving the thermostability of a previously found firefly Hotaria parvula luciferase mutant with minor pH-sensitive spectral change (V368A), further mutation (E356R) was introduced by taking a reportedly stabilized mutant of Photinus pyralis luciferase into account. The double mutant E356R/V368A showed significantly improved thermostability because > 90% activity remained after incubation for 1 h at 45 degrees C, with its specific activity being maintained. Unlike the wild type or V368A, E356R/V368A showed no change in the emission maximum of 568 nm even at pH 6.3, also implying a mutual relationship between thermostability and the proportion of yellow-green luminescent peak under acidic condition.


Evaluating cell viability, capillary perfusion, and collateral tortuosity in an ex vivo mouse intestine fluidics model.
Numerous disease conditions involve the sudden or progressive loss of blood flow. Perfusion restoration is vital for returning affected organs to full health. While a range of clinical interventions can successfully restore flow to downstream tissues, the microvascular responses after a loss-of-flow event can vary over time and may involve substantial microvessel instability. Increased insight into perfusion-mediated capillary stability and access-to-flow is therefore essential for advancing therapeutic reperfusion strategies and improving patient outcomes. To that end, we developed a tissue-based microvascular fluidics model to better understand (i) microvascular stability and access-to-flow over an acute time course post-ischemia, and (ii) collateral flow in vessels neighboring an occlusion site. We utilized murine intestinal tissue regions by catheterizing a feeder artery and introducing perfusate at physiologically comparable flow-rates. The cannulated vessel as well as a portion of the downstream vessels and associated intestinal tissue were cultured while constant perfusion conditions were maintained. An occlusion was introduced in a selected arterial segment, and changes in perfusion within areas receiving varying degrees of collateral flow were observed over time. To observe the microvascular response to perfusion changes, we incorporated (i) tissues harboring cell-reporter constructs, specifically Ng2-DsRed labeling of intestinal pericytes, and (ii) different types of fluorescent perfusates to quantify capillary access-to-flow at discrete time points. In our model, we found that perfusion tracers could enter capillaries within regions downstream of an occlusion upon the initial introduction of perfusion, but at 24 h tissue perfusion was severely decreased. However, live/dead cell discrimination revealed that the tissue overall did not experience significant cell death, including that of microvascular pericytes, even after 48 h. Our findings suggest that altered flow conditions may rapidly initiate cellular responses that reduce capillary access-to-flow, even in the absence of cellular deterioration or hypoxia. Overall, this ex vivo tissue-based microfluidics model may serve as a platform upon which a variety of follow-on studies may be conducted. It will thus enhance our understanding of microvessel stability and access-to-flow during an occlusive event and the role of collateral flow during normal and disrupted perfusion.


DNA ligases ensure fidelity by interrogating minor groove contacts.
DNA ligases, found in both prokaryotes and eukaryotes, covalently link the 3'-hydroxyl and 5'-phosphate ends of duplex DNA segments. This reaction represents a completion step for DNA replication, repair and recombination. It is well established that ligases are sensitive to mispairs present on the 3' side of the ligase junction, but tolerant of mispairs on the 5' side. While such discrimination would increase the overall accuracy of DNA replication and repair, the mechanisms by which this fidelity is accomplished are as yet unknown. In this paper, we present the results of experiments with Tth ligase from Thermus thermophilus HB8 and a series of nucleoside analogs in which the mechanism of discrimination has been probed. Using a series of purine analogs substituted in the 2 and 6 positions, we establish that the apparent base pair geometry is much more important than relative base pair stability and that major groove contacts are of little importance. This result is further confirmed using 5-fluorouracil (FU) mispaired with guanine. At neutral pH, the FU:G mispair on the 3' side of a ligase junction is predominantly in a neutral wobble configuration and is poorly ligated. Increasing the solution pH increases the proportion of an ionized base pair approximating Watson-Crick geometry, substantially increasing the relative ligation efficiency. These results suggest that the ligase could distinguish Watson-Crick from mispaired geometry by probing the hydrogen bond acceptors present in the minor groove as has been proposed for DNA polymerases. The significance of minor groove hydrogen bonding interactions is confirmed with both Tth and T4 DNA ligases upon examination of base pairs containing the pyrimidine shape analog, difluorotoluene (DFT). Although DFT paired with adenine approximates Watson-Crick geometry, a minor groove hydrogen bond acceptor is lost. Consistent with this hypothesis, we observe that DFT-containing base pairs inhibit ligation when on the 3' side of the ligase junction. The NAD+-dependent ligase, Tth, is more sensitive to the DFT analog on the unligated strand whereas the ATP-dependent T4 ligase is more sensitive to substitutions in the template strand. Electrophoretic gel mobility-shift assays demonstrate that the Tth ligase binds poorly to oligonucleotide substrates containing analogs with altered minor groove contacts.


Antibodies against I-A and I-E determinants inhibit the activation and function of encephalitogenic T-lymphocyte lines.
Two monoclonal antibodies, OX-6 and OX-17, were used to evaluate respectively the roles of I-A and I-E major histocompatibility complex Class II gene products in the in vitro activation and subsequent function in recipient rats of encephalitogenic T-cell lines. Activation of the T-cell lines with guinea pig myelin basic protein (GP-BP) presented by accessory cells (APC) resulted in an increase in the number of blast cells in culture and was reflected by increased uptake of [3H]thymidine [( 3H]Tdy). The number of blasts recovered and [3H]Tdy uptake during activation was reduced drastically in the presence of OX-6, but to a much lesser extent in the presence of OX-17. OX-6 but not OX-17 appeared to block T-cell activation primarily by inhibiting APC function, since preincubation of APC but not T cells with OX-6 before stimulation resulted in complete inhibition of the cultures. After activation, the BP-1 T-cell line or D-9 clone transferred severe paralysis to normal recipient rats. Recipients of OX-6-treated BP-1 or D-9 T cells exhibited very mild or no signs, whereas recipients of OX-17-treated cells developed only slightly less severe experimental autoimmune encephalomyelitis (EAE) than recipients of untreated encephalitogenic control cultures. In contrast, treatment with OX-17 but not OX-6 reduced the ability of BP-reactive T cells to transfer delayed-type hypersensitivity reactions. Dermal testing with GP-BP in the ears of recipient rats just prior to onset of clinical signs decreased significantly the clinical intensity of EAE induced by activated BP-reactive T cells, but increased the clinical scores in rats which received unstimulated or OX-6-treated T cells. This potentiating effect of GP-BP was due most likely to the presentation of processed antigen to circulating BP-reactive T cells by APC in the ear. These results suggest that both the I-A and I-E gene products may contribute to the activation and subsequent function of encephalitogenic T cells, perhaps through separate mechanisms.


Inhibitory effects of high temperature- and pressure-treated garlic on formation of 1,2-dimethylhydrazine-induced mucin-depleted foci and O(6)-methylguanine DNA adducts in the rat colorectum.
High temperature- and pressure-treated garlic (HTPG) has been reported to have enhanced antioxidative and cytotoxic activities. However, there have been no reports on chemopreventive effects using animal cancer models. This study first examined the modifying effects of HTPG on 1,2-dimethylhydrazine (DMH)-induced mucin-depleted foci (MDF) and aberrant crypt foci (ACF), preneoplastic lesions in the rat colorectum. Male F344 rats (5 weeks old) were fed basal diet, or experimental diets containing 1% or 3% HTPG for 5 weeks. One week later, all rats were injected s.c.with DMH (40 mg/kg, once weekly for 2 weeks). At 10 weeks of age, all the rats were sacrificed, and the colorectum was evaluated for MDF and ACF. In rats given DMH and 3% HTPG, the numbers of MDF were decreased significantly as compared with those of rats given DMH alone (p< 0.01), and the numbers of ACF showed a tendency to decrease, although not significantly. Next, the effects of HTPG on the formation of DMH-induced O6-methylguanine (O6-MeG) DNA adducts in rats were studied. Male F344 rats (5 weeks old) were fed the basal diet or 10% HTPG diet for 5 weeks. All rats were injected i.p. once with 40 mg/kg DMH at the end of week 5. The animals were sacrificed 6 hours after DMH injection to analyze the O6-MeG DNA adducts in the colorectal mucosa and liver. Dietary administration of HTPG significantly reduced the adduct levels in the colorectal mucosa and liver, compared with the controls (both p< 0.01). The activities of some detoxification enzymes in the liver of DMH-treated rats were also measured. HTPG significantly reduced the activity of cytochrome P450 (CYP) 2E1, known to be responsible for activation of DMH in rat liver (p< 0.05). In contrast, HTPG significantly enhanced the activities of phase 2 enzymes, quinone reductase (QR) and glutathione S-transferase (GST), in rat liver (both p< 0.05). These results suggested that HTPG might have chemopreventive effects against colon carcinogenesis, at least in the initiation stage.


Identification of Inhibitors of Integrin Cytoplasmic Domain Interactions With Syk.
Leukocyte inflammatory responses require integrin cell-adhesion molecule signaling through spleen tyrosine kinase (Syk), a non-receptor kinase that binds directly to integrin β-chain cytoplasmic domains. Here, we developed a high-throughput screen to identify small molecule inhibitors of the Syk-integrin cytoplasmic domain interactions. Screening small molecule compound libraries identified the β-lactam antibiotics cefsulodin and ceftazidime, which inhibited integrin β-subunit cytoplasmic domain binding to the tandem SH2 domains of Syk (IC50 range, 1.02-4.9 µM). Modeling suggested antagonist binding to Syk outside the pITAM binding site. Ceftazidime inhibited integrin signaling via Syk, including inhibition of adhesion-dependent upregulation of interleukin-1β and monocyte chemoattractant protein-1, but did not inhibit ITAM-dependent phosphorylation of Syk mediated by FcγRI signaling. Our results demonstrate a novel means to target Syk independent of its kinase and pITAM binding sites such that integrin signaling via this kinase is abrogated but ITAM-dependent signaling remains intact. As integrin signaling through Syk is essential for leukocyte activation, this may represent a novel approach to target inflammation.


Comparison between MRI and the Combination of 2D and 3D US in the Prenatal Diagnosis of Closed Spina Bifida.
Closed spina bifida (CSB) is a rare condition with a challenging prenatal diagnosis. Herein, we assess the conventional two-dimensional (2D) ultrasound (US) combined with three-dimensional (3D) ultrasound (US) and magnetic resonance imaging (MRI) in the prenatal diagnosis of CSB. In this retrospective study, we included 20 cases of fetal CSB confirmed by postnatal MRI, post-mortem pathological examination, or postpartum surgery. Prenatal 2D US complemented with the 3D US was performed in all fetuses to evaluate the characteristics of the conus, vertebral arch, and scoliosis. Moreover, MRI was performed to establish the split vertebrae, with or without a bulging mass. Thereafter, we compared the performance of the US and MRI. Diagnosis accuracy of US was comparable with MRI (70% vs. 75%, κ = 0.62); US detected more cases with interpediculate distance ≥95% (55% vs. 35%, κ = 0.22) than MRI. On the other hand, MRI had a superior capacity for identifying vertebral arch fissures (20% vs. 35%, κ = 0.39). MRI and ultrasound had good agreement in the conus medullaris (65% vs. 70%, κ = 0.42) and scoliosis (45% vs. 35%, κ = 0.59). Both US and MRI detected 1 (5.0%) case with "lemon sign" and "banana sign." The missed diagnosis rates of US and MRI were 15% (3/20) and 5% (1/20), respectively. The misdiagnosis rates of US and MRI were 15.0% (3/20) and 20.0% (4/20), respectively. Both MRI and 2D US combined with the 3D US had excellent performance in prenatal diagnosis of CSB.


[Study of spatial organization of the RNA of phage MSZ using nucleases specific for the secondary structure].
The limited hydrolisis of bacteriophage MS2 RNA by nuclease S1 and ds-specific snake venom RNase was studied in a wide range of ionic strength, at different pH, after heating and (slow and fast) cooling and at various enzyme-substrate relations. It was shown that the RNA has exposed hydrolisis sites for both nucleases. The localizations of these sites are very specific and are not altered in all conditions studied. The hydrolisis rate was changed in some conditions, at that the fragments patterns in denaturing electrophoresis did not move. It was supposed that the RNA has strongly predetermined and predominant conformation which could not be altered by strong influences.


Reliability of phase-specific outcome measurements in change-of-direction tests using a motorized resistance device.
This study aims to determine test-retest reliability of phase-specific information during initial acceleration, deceleration, and re-acceleration phases of different change-of-direction (CoD) tests using a motorized resistance device (MRD). A total of 21 participants (16 males and five females, with mean age of 22.3 ± 3.9 years, body mass of 75.2 ± 6.9 kg, height of 177.9 ± 6.8 cm) completed the modified 505 (m505), 10-0-5, and 15-0-5 CoD tests on four different test sessions while exposed to an external load (3 kg) provided by the MRD. Outcome variables included overall and phase-specific kinetic (force, power, and impulse) and kinematic (time, distance, velocity, and acceleration/deceleration) data during the initial acceleration, deceleration, and re-acceleration phases. The deceleration and re-acceleration phases were further divided into two subphases, namely, early and late subphases, using 50% of maximum velocity. Reliability was assessed using an intraclass correlation coefficient (ICC), coefficient of variation (CV), typical error (TE), and minimal detectable change (MDC). Good to excellent ICC values (>0.75) and acceptable (<10%) to good (<5%) CV values were observed for most outcome measurements. Specifically, 80.1% (822 out of 1,026) of all variables showed good or better relative reliability (i.e., ICC ≥ 0.75), while 97.0% (995 out of 1,026) of all variables showed acceptable or better absolute reliability (i.e., CV < 10%). In conclusion, the present study demonstrates that the MRD can obtain reliable phase-specific outcome measurements across different CoD tests, providing coaches and researchers with new opportunities to advance our understanding of CoD ability and inform more advanced CoD training prescriptions.


A novel protective function for cytokinin in the light stress response is mediated by the Arabidopsis histidine kinase2 and Arabidopsis histidine kinase3 receptors.
Cytokinins are plant hormones that regulate diverse processes in plant development and responses to biotic and abiotic stresses. In this study, we show that Arabidopsis (Arabidopsis thaliana) plants with a reduced cytokinin status (i.e. cytokinin receptor mutants and transgenic cytokinin-deficient plants) are more susceptible to light stress compared with wild-type plants. This was reflected by a stronger photoinhibition after 24 h of high light (approximately 1,000 µmol m(-2) s(-1)), as shown by the decline in maximum quantum efficiency of photosystem II photochemistry. Photosystem II, especially the D1 protein, is highly sensitive to the detrimental impact of light. Therefore, photoinhibition is always observed when the rate of photodamage exceeds the rate of D1 repair. We demonstrate that in plants with a reduced cytokinin status, the D1 protein level was strongly decreased upon light stress. Inhibition of the D1 repair cycle by lincomycin treatment indicated that these plants experience stronger photodamage. The efficiency of photoprotective mechanisms, such as nonenzymatic and enzymatic scavenging systems, was decreased in plants with a reduced cytokinin status, which could be a cause for the increased photodamage and subsequent D1 degradation. Additionally, slow and incomplete recovery in these plants after light stress indicated insufficient D1 repair. Mutant analysis revealed that the protective function of cytokinin during light stress depends on the Arabidopsis histidine KINASE2 (AHK2) and AHK3 receptors and the type B Arabidopsis response regulator1 (ARR1) and ARR12. We conclude that proper cytokinin signaling and regulation of specific target genes are necessary to protect leaves efficiently from light stress.


Mechanism by which ammonium bicarbonate and ammonium sulfate inhibit mycotoxigenic fungi.
In this study we examined the mechanism by which ammonium bicarbonate inhibits mycotoxigenic fungi. Elevated extracellular pH, alone, was not responsible for the antifungal activity. Although conidia of Penicillium griseofulvum and Fusarium graminearum had internal pH (pHi) values as high as 8.0 in buffer at an external pH (pHo) of 9.5, their viability was not markedly affected. The pHi values from conidia equilibrated in glycine-NaOH-buffered treatments without ammonium bicarbonate or ammonium sulfate were similar to values obtained from buffered treatments containing the ammonium salts. Thus, inhibition did not appear to be directly related to increased pHi. Ammonium sulfate in buffered media at pH greater than or equal to 8.7 was as inhibitory as ammonium bicarbonate, but was completely ineffective at pH less than or equal to 7.8. The hypothesis that free ammonia caused the fungal inhibition was tested by using ammonium sulfate as a model for ammonium bicarbonate. Viability, expressed as log CFU/ml, and percent germination of P. griseofulvum and F. graminearum decreased dramatically as the free ammonia concentration increased. Germination rate ratios (the germination rate in buffered ammonium sulfate divided by the germination rate in buffer alone) decreased linearly as the free ammonia concentration increased, further establishing NH3 as the toxic agent. Ammonium bicarbonate inhibits fungi because the bicarbonate anion supplies the alkalinity necessary to establish an antifungal concentration of free ammonia.


Diet in Early Pregnancy: Focus on Folate, Vitamin B12, Vitamin D, and Choline.
Purpose: Prenatal multivitamins are recommended in pregnancy. This study assessed food and supplement intakes of folate, vitamin B12 (B12), vitamin D, and choline in pregnant women living in Southern Ontario in comparison with current recommendations. Methods: Women recruited to the Be Healthy in Pregnancy RCT (NCT01693510) completed 3-day diet/supplement records at 12-17 weeks gestation. Intakes of folate, B12, vitamin D, and choline were quantified and compared with recommendations for pregnant women. Results: Folate intake (median (min, max)) was 1963 μg/day dietary folate equivalents (153, 10 846); 90% of women met the Estimated Average Requirement (EAR) but 77% exceeded the Tolerable Upper Intake Level (UL) (n = 232). B12 intake was 12.1 μg/day (0.3, 2336); 96% of women met the EAR with 7% exceeding the EAR 100-fold (n = 232). Vitamin D intake was 564 IU/day (0.0, 11 062); 83% met the EAR, whereas 1.7% exceeded the UL (n = 232). Choline intake was 338 mg/day (120, 1016); only 18% met the Adequate Intake and none exceeded the UL (n = 158). Conclusion: To meet the nutrient requirements of pregnancy many women rely on prenatal vitamins. Reformulating prenatal multivitamin supplements to provide doses of vitamins within recommendations to complement a balanced healthy diet would ensure appropriate micronutrient intakes for pregnant women.


Triplet gestation: clinical outcome of 14 cases.
To determine maternal complications and fetal outcome of triplet gestations. Retrospective study of pregnant women with triplet gestation managed in 10 years. Fourteen women were managed with triplet gestation, of these, (71.4%) were booked for antenatal care and four (28.6%) were unbooked. The mean age of the women was 31.3 years. The age range was between twenty seven years and thirty nine years. The mean gestational age at diagnosis for the booked women was 18.6 weeks. Of the fourteen patients, ten (71.4%) had spontaneous conception, three (21.4%) followed ovulation induction and one (7.2%) resulted from invitro fertilization and embryo transfer. Two (14.3%) patients had cervical cerclage based on their past obstetric history and assessment of the cervix. Six (42.9%) patients were hospitalized and treated for preeclampsia 3 patients, spontaneous abortion 1 patient and cervical incompetence 2 patients. Eleven (78.6%) patients had preterm birth. The mean gestational age at delivery was 33.4 weeks. Of the thirteen deliveries, nine (69.2%) had caesarean section and four (30.8%) delivered per vaginam. A total of thirty nine babies were delivered, thirty four (87.2%) babies survived and five (12.8%) died. Perinatal mortality was 11.9% and the "take home" baby rate was 81%. Antenatal care with initiation of specialized prenatal care and planned delivery in triplet gestation improves fetal outcome.


Chinese Public's Attention to the COVID-19 Epidemic on Social Media: Observational Descriptive Study.
Since the coronavirus disease (COVID-19) epidemic in China in December 2019, information and discussions about COVID-19 have spread rapidly on the internet and have quickly become the focus of worldwide attention, especially on social media. This study aims to investigate and analyze the public's attention to events related to COVID-19 in China at the beginning of the COVID-19 epidemic (December 31, 2019, to February 20, 2020) through the Sina Microblog hot search list. We collected topics related to the COVID-19 epidemic on the Sina Microblog hot search list from December 31, 2019, to February 20, 2020, and described the trend of public attention on COVID-19 epidemic-related topics. ROST Content Mining System version 6.0 was used to analyze the collected text for word segmentation, word frequency, and sentiment analysis. We further described the hot topic keywords and sentiment trends of public attention. We used VOSviewer to implement a visual cluster analysis of hot keywords and build a social network of public opinion content. The study has four main findings. First, we analyzed the changing trend of the public's attention to the COVID-19 epidemic, which can be divided into three stages. Second, the hot topic keywords of public attention at each stage were slightly different. Third, the emotional tendency of the public toward the COVID-19 epidemic-related hot topics changed from negative to neutral, with negative emotions weakening and positive emotions increasing as a whole. Fourth, we divided the COVID-19 topics with the most public concern into five categories: the situation of the new cases of COVID-19 and its impact, frontline reporting of the epidemic and the measures of prevention and control, expert interpretation and discussion on the source of infection, medical services on the frontline of the epidemic, and focus on the worldwide epidemic and the search for suspected cases. Our study found that social media (eg, Sina Microblog) can be used to measure public attention toward public health emergencies. During the epidemic of the novel coronavirus, a large amount of information about the COVID-19 epidemic was disseminated on Sina Microblog and received widespread public attention. We have learned about the hotspots of public concern regarding the COVID-19 epidemic. These findings can help the government and health departments better communicate with the public on health and translate public health needs into practice to create targeted measures to prevent and control the spread of COVID-19.


Scalable solid-template reduction for designed reduced graphene oxide architectures.
Herein, we report a solid-state reduction process (in contrast to solution-based approach) by using an environmentally friendly reductant, such as vitamin C (denoted VC), to be directly employed to solid-state graphene oxide (GO) templates to give the highly active rGO architecture with a sheet resistance of as low as 10 Ω sq(-1). In addition, predesigned rGO patterns/tracks with tunable resistivity can be directly "written" on a preprepared solid GO film via the inkjet-printing technique using VC/H2O as the printing-ink. This advanced reduction process allows foreign active materials to be preincorporated into the GO matrix to form quality active composite architectures.


Production of Egg Yolk Antibody (IgY) Against Vibrio Cholerae O1: Protective Effect in Mice.
Cholera is an acute intestinal infection caused by Vibrio cholera (V. cholera). The development of antibodies against specific V. cholerae may have a therapeutic effect. In the present research, we investigated the protective effect of egg yolk Immunoglobulin (IgY), which was produced by immunizing hens with formaldehyde-killed V. cholerae O1 and subsequently the isolated IgY was orally administrated to the V. cholerae O1 infected mice for evaluation of its immunizing capability. In the current study, hens were immunized three times with formaldehyde-killed V. cholerae O1 (1.5×107 CFU/ml) and an equal volume of adjuvant. The IgY was isolated from egg yolk by polyethylene glycol method. The validity and activity of isolated IgY were confirmed with SDS-PAGE and ELISA methods, respectively. Subsequently IgY was orally administered to suckling mice following challenge with V. cholerae O1. ELISA results showed high antibody titer in the serum and egg yolk. Also, SDS-PAGE analysis showed successful purification of IgY and anti-V. cholerae IgY prevented the death of mice infected with V. cholerae O1. The anti-V. cholera IgY was administered at 2, 4, 6 hr intervals after 3 hr of inoculation of mice with V. cholerae O1. Results showed that the rate of surviving mice (2 mg/ml of IgY) were 60% after 4 hr and 40% after 6 hr and the rate of surviving mice (5 mg/ml of IgY) was 70% after 4 hr and 60% after 6 hr. The findings suggested the egg yolk-driven IgY as a natural antibacterial protein, could be effective in the prevention and treatment of cholera disease.


Global genome and transcription-coupled nucleotide excision repair pathway in prokaryotes.
In all cells-from bacteria to humans-nucleotide excision repair (NER) is a highly conserved, versatile DNA repair pathway that is responsible for the removal of a wide variety of DNA helix-distorting lesions arising from both endogenous and exogenous sources. In many organisms including bacteria, fungi, animals, and plants, NER occurs through two sub-pathways: the global genome NER (GG-NER) pathway and the transcription- coupled NER (TC-NER) pathway. Although essential factors and basic steps involved in NER have been identified, mechanisms and stages of their assembly process are not well understood. In this review, we summarize recent literature about protein interaction networks that manifest during initial stages of bacterial NER pathway while highlighting some of the key functional studies.


Three score and ten.
Excluding infant and child mortality, the average "natural" life span seems to be about 70 years. It has remained so for several thousand years. It shows little sign of change. Hard work and stress appear to increase rather than decrease longevity.


Non-invasive cardiac output monitoring for cesarean delivery under epidural anesthesia in a patient with Marfan syndrome and cardiomyopathy.
Maternal cardiac output and stroke volume increase significantly at the time of cesarean delivery. Parturients with baseline myocardial dysfunction are at increased risk of cardiovascular decompensation in the peripartum period and close hemodynamic monitoring is warranted. We report our use of intraoperative non-invasive cardiac output monitoring during cesarean delivery under epidural anesthesia in a 24-year-old woman with dilated cardiomyopathy secondary to Marfan syndrome, aortic arch, aortic valve and mitral valve replacements and a left ventricular ejection fraction of 37%. Three distinct hemodynamic trends were noted. After achieving adequate surgical anesthesia with 2% lidocaine 20mL, cardiac output and stroke volume rose for approximately 20min from baseline values of 6.3L/min and 69mL, respectively, to 9L/min and 107mL. Values subsequently trended down and remained depressed for nearly 20min following delivery. The lack of immediate post-delivery increases in both cardiac output and stroke volume were attributed to acute blood loss, intravascular volume depletion from fluid restriction, and slow infusion of oxytocin. By the end of surgery, cardiac output and stroke volume ultimately increased by 66% and 84% of baseline values, respectively. Systemic blood pressure, heart rate and cardiac output did not appear to correlate despite the use of phenylephrine to manage hypotension. The patient remained hemodynamically stable with no evidence of acute volume overload.


Detection of 28 neurotransmitters and related compounds in biological fluids by liquid chromatography/tandem mass spectrometry.
This work presents two liquid chromatography/tandem mass spectrometry (LC/MS/MS) acquisition modes: multiple reaction monitoring (MRM) and neutral loss scan (NL), for the analysis of 28 compounds in a mixture. This mixture includes 21 compounds related to the metabolism of three amino acids: tyrosine, tryptophan and glutamic acid, two pterins and five deuterated compounds used as internal standards. The identification of compounds is achieved using the retention times (RT) and the characteristic fragmentations of ionized compounds. The acquisition modes used for the detection of characteristic ions turned out to be complementary: the identification of expected compounds only is feasible by MRM while expected and unexpected compounds are detected by NL. In the first part of this work, the fragmentations characterizing each molecule of interest are described. These fragmentations are used in the second part for the detection by MRM and NL of selected compounds in mixture with and without biological fluids. Any preliminary extraction precedes the analysis of compounds in biological fluids.


Interpretation of vaccine associated neurological adverse events: a methodological and historical review.
As a result of significant recent scientific investment, the range of vaccines available for COVID-19 prevention continues to expand and uptake is increasing globally. Although initial trial safety data have been generally reassuring, a number of adverse events, including vaccine induced thrombosis and thrombocytopenia (VITT), have come to light which have the potential to undermine the success of the vaccination program. However, it can be difficult to interpret available data and put these into context and to communicate this effectively. In this review, we discuss contemporary methodologies employed to investigate possible associations between vaccination and adverse neurological outcomes and why determining causality can be challenging. We demonstrate these issues by discussing relevant historical exemplars and explore the relevance for the current pandemic and vaccination program. We also discuss challenges in understanding and communicating such risks to clinicians and the general population within the context of the 'infodemic' facilitated by the Internet and other media.


Operative vs percutaneous drainage of intra-abdominal abscesses. Comparison of morbidity and mortality.
This retrospective case-controlled study compares the morbidity and mortality of 27 percutaneously drained (PD) abscesses with 27 that were surgically drained (SD). Patients were matched for age, sex, diagnosis, and abscess etiology and location. There was no difference in severity of illness (acute physiology score [APS] = 8.3 vs 10.2), comparable morbidity (29.6% vs 40.7%), or mortality (11.0% vs 7.4%) between PD and SD groups. Duration of drainage was significantly longer in the PD group; however, this is explained in part by the 48% vs 18.5% difference in associated fistulae. Failures of the SD group had a higher mean APS (15) than both failures of the PD group (APS = 9.3) and successes of the SD group (APS = 8.6). All three PD group deaths and half of the SD group deaths were related to ongoing sepsis. Surgical drainage of intra-abdominal abscess is as successful as PD. Percutaneous drainage is reasonable initial treatment for intra-abdominal abscess; however, early assessment of clinical status and frequent reassessment are mandatory to ensure that failures are dealth with early. We present a drainage algorithm.


[Odontogenesis, organogenic model using teratomas].
The study of odontogenesis in teratomas, leads to the following observations. Conspicuous odontogenic fields are present in the vicinity of pluri-differentiated cystic structures. Tooth germs are connected to the walls of the cysts by an anastomosed dental laminar. The teeth developing in teratoma are not comparable to the normal process which is harmonized when the formation and the distribution of the various parts are concerned. They are deformed in appearance but not in the same way as the classic genetic malformations. The origin of both epithelial and mesenchymal parts is still not completely understood.


[Vascular complications associated with inflammatory bowel disease].
Thromboembolic episode is a well known extraintestinal manifestation of inflammatory bowel disease, but it is a clinical rare complication. Histological and hematological studies suggest that a hypercoagulable state is involved in the pathogenesis of inflammatory bowel disease. However, the exact mechanism of hypercoagulability is still unknown. During the acute recurrences there is an increase of factor VIII, fibrinogen, platelet, factor V and decrease of antithrombin III. Hematologic disorders seem markedly correlated with the activity of the disease. We report on two patients with Inflammatory bowel disease and hypercoagulable state. We review the literature and discuss about the pathogenic mechanisms of such complication.


Ca paradox in neural injury: a hypothesis.
The deleterious effects of Ca ionic entry into neurons has been speculated to be a final common pathway of cell death. However, a direct cause-effect relationship between Ca and neuronal death has been difficult to establish. Cells dying from any cause will accumulate Ca. The entry of Ca into neurons and the subsequent pathological changes associated with Ca entry consequently may be manifestations rather than causes of cell death. Recent work showing that extracellular Ca ionic activity becomes profoundly depressed in injured spinal cord and ischemic cerebral cortex prompted a new hypothesis on Ca mediated damage. We propose that the initial fall in extracellular Ca activity, resulting from the death of some cells in the tissue, increases the susceptibility of the surviving cells to Ca entry when extracellular Ca activity levels normalize and that this accounts for part of the secondary damage that has been observed in neural injury models. Such a phenomenon has been described in cardiac tissues. Dubbed Ca paradox, this phenomenon occurs when heart cells are perfused with Ca-free solutions for several minutes followed by the return to normal Ca-containing solutions. The cardiac cells die and undergo physiological, morphological, biochemical, and other changes. The evidence supporting a Ca paradox phenomenon in injured neural tissues is summarized. The therapeutic implications of Ca paradox in neural tissue injury are discussed.


Prosocial effects of prolactin in male rats: Social recognition, social approach and social learning.
Prolactin (PRL) and oxytocin (OT) are pituitary hormones essential for lactation, but also promote sexual behavior. OT stimulates social behaviors, such as recognition, approach, and learning, but less is known about PRL in these behaviors. Since PRL and OT have complementary functions in reproduction, we hypothesized that PRL increases social recognition, approach, and learning. Male Long-Evans rats received ovine PRL (oPRL; 0.5, 2.0 or 5.0mg/kg), the PRL antagonist bromocriptine (0.1, 3.0 or 5.0mg/kg) or saline 20 mins before testing for recognition of familiar vs. unfamiliar stimulus males. Saline controls preferred the unfamiliar male (p<0.05), while bromocriptine blocked this preference. oPRL did not increase preference. To measure social approach, we determined if PRL restores approach 2h after defeat by an aggressive male. Defeated rats avoided the aggressive male. 2mg/kg oPRL, before or after defeat, restored approach towards the aggressive male (p<0.05). In non-defeated rats, oPRL or 3mg/kg bromocriptine had no effect. To determine if PRL increases social learning, we tested social transmission of food preference. Rats choose between two unfamiliar flavors, one of which they have previously been exposed to through interaction with a demonstrator rat. Vehicle controls preferred chow with the demonstrated flavor over the novel flavor. oPRL-treated rats were similar. Bromocriptine-treated rats failed to show a preference. When tested one week later, only oPRL-treated rats preferred the demonstrated flavor. The results suggest that PRL is required for social recognition and learning, and that increasing PRL enhances social memory and approach, similar to OT.


The effect of manganese on lipogenesis, lipolysis, adenosine 3',5' cyclic monophosphate and cyclic nucleotide phosphodiesterase activity in adipocytes from NZY mice.
The insulin-like action of Mn2+ was investigated in adipocytes isolated from male mice of the NZY strain. In agreement with previous reports Mn2+ was found to stimulate both the oxidation of [U-14C]glucose to CO2 and the incorporation of [U-14C]glucose into total lipid and fatty acid, and to inhibit lipolysis stimulated by epinephrine, cyclic AMP or theophylline. The maximum effect of Mn2+ was greater than that of a maximal concentration of insulin and when both agents were present in these concentrations the effect was similar to that observed with Mn2+ alone. Mn2+ lowered the level of cyclic AMP in adipocytes incubated with isoproterenol. The effect was seen as early as 1 minute and it was greater than a maximal concentration of insulin. When Mn2+ was added to suspensions of adipocytes it increased the activity of the membrane-bound low Km cyclic nucleotide phosphodiesterase in subsequently prepared homogenates. The enzyme was stimulated 1.8-fold by Mn2+ compared with a 1.7-fold stimulation by insulin and a 2-fold stimulation in the presence of both Mn2+ and insulin.


Art is Patient: A Museum-Based Experience to Teach Trauma-Sensitive Engagement in Health Care.
Psychological trauma is ubiquitous, an often hidden yet influential factor in care across clinical specialties. Interdisciplinary health professions education is mobilizing to address the importance of trauma-sensitive care. Given their attention to complex human realities, the health humanities are well-poised to shape healthcare learners' responses to trauma. Indeed, many such arts and humanities curricula propose narrative exercises to strengthen empathy, self-reflection, and sensitive communication. Trauma, however, is often unwordable, fragmentary, and physically encoded, incompatible with storying methods. This article presents a recent innovation, the Art is Patient seminar series, which focuses on aesthetic exercises to help learners access and share non-verbal, embodied, and relational responses to art. Based in an art museum context, it provides successive experiences of approaching, witnessing, and engaging with visual art as an analogue to developing trauma-sensitive relationships. Reflections on the process locate the seminar vis-à-vis health humanities practices, aesthetics, and trauma-informed approaches.


Proteomic Profiling of Extracellular Vesicles Separated from Plasma of Former National Football League Players at Risk for Chronic Traumatic Encephalopathy.
Chronic Traumatic Encephalopathy (CTE) is a tauopathy that affects individuals with a history of exposure to repetitive head impacts, including National Football League (NFL) players. Extracellular vesicles (EVs) are known to carry tau in Alzheimer's disease and other tauopathies. We examined protein profiles of EVs separated from the plasma of former NFL players at risk for CTE. EVs were separated from the plasma from former NFL players and age-matched controls using size-exclusion chromatography. Label-free quantitative proteomic analysis identified 675 proteins in plasma EVs, and 17 proteins were significantly differentially expressed between former NFL players and controls. Total tau (t-tau) and tau phosphorylated at threonie181 (p-tau181) in plasma-derived EVs were measured by ultrasensitive immunoassay. Level of t-tau and p-tau181 in EVs were significantly different, and the area under the receiver operating characteristic curve (AUC) of t-tau and p-tau181 showed 0.736 and 0.715, respectively. Machine learning analysis indicated that a combination of collagen type VI alpha 3 and 1 chain (COL6A3 and COL6A1) and reelin (RELN) can distinguish former NFL players from controls with 85% accuracy (AUC = 0.85). Based on the plasma EV proteomics, these data provide protein profiling of plasma EVs for CTE, and indicate combination of COL6A3, RELN and COL6A1 in plasma EVs may serve as the potential diagnostic biomarkers for CTE.


Enoxacin for the treatment of urinary tract infection.
Enoxacin, a new orally absorbed broad spectrum anti-microbial agent, was used to treat 25 patients with moderate or severe urinary tract infections. Many were complicated and associated with renal insufficiency. Eighteen of the 25 were cured, five had a relapse and two became reinfected with a resistant Streptococcus faecalis. Fourteen of the patients reported side effects, including six with a neurological syndrome, one with widespread arthralgia and one with an Achilles tendonitis. Further studies are indicated with this potent drug to assess its role in the management of severe or complicated urinary infections and to assess the optimum dose and duration of therapy.


Pharmacological discrimination of GluR5 and GluR6 kainate receptor subtypes by (3S,4aR,6R,8aR)-6-[2-(1(2)H-tetrazole-5-yl)ethyl]decahyd roisdoquinoline-3 carboxylic-acid.
The pharmacological tools available for the discrimination of kainate receptor subtypes are limited. We examined the effects of (3S,4aR,6R,8aR)-6-[2-(1(2)H-tetrazole-5-yl)ethyl]decahydr oisoquinoline-3-carboxylic acid (LY293558) and 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline (NBQX) on inward currents associated with activation of non-N-methyl-D-asparate (NMDA) receptors in acutely isolated rat cerebellar Purkinje neurons, rat dorsal root ganglion neurons, and human embryonic kidney 293 cells transfected with human glutamate receptors (GluR) 5 and 6. LY293558 and NBQX inhibited kainate-induced currents in cerebellar Purkinje cells, DRG neurons, and human GluR5-transfected cells. In contrast, human embryonic kidney 293 cells expressing GluR6 receptors, although blocked by NBQX, were unaffected by LY293558 at concentrations of < / = 100 microM. The selective antagonism by LY293558 of GluR5 receptors should allow the determination of the functional role of GluR5 and GluR6 in more complex systems.


[Methods of Assessment of the Right Ventricular Function].
The review contains contemporary data of assessment on right ventricular (RV) function by various instrumental methods of investigation. Among all these methods multispiral computed tomography (MSCT) appears to be most perspective. Accuracy and reproducibility of measurements of RV function by MSCT and comparable with those of magnetic resonance tomography. The uniqueness of MSCT is its ability to produce complex simultaneous evaluation not only of RV-functional parameters but also of other anatomical structures located in the scanning field.


Apolipoprotein E3/E3 genotype decreases the risk of pituitary dysfunction after traumatic brain injury due to various causes: preliminary data.
Traumatic brain injury (TBI) is a devastating public health problem which may result in hypopituitarism. However, the mechanisms and the risk factors responsible for hypothalamo-pituitary dysfunction due to TBI are still unclear. Although APO E is one of the most abundant protein in hypothalamo-pituitary region, there is no study investigating the relation between APO E polymorphism and TBI-induced hypopituitarism. This study was undertaken to determine whether APO E genotypes modulate the pituitary dysfunction risk after TBI due to various causes, including traffic accident, boxing, and kickboxing. Ninety-three patients with TBI (mean age, 30.61 +/- 1.25 years) and 27 healthy controls (mean age, 29.03 +/- 1.70 years) were included in the study. Pituitary functions were evaluated, and APO E genotypes (E2/E2; E3/E3; E4/E4; E2/E3; E2/E4; E3/E4) were screened. Twenty-four of 93 subjects (25.8%) had pituitary dysfunction after TBI. The ratio of pituitary dysfunction was significantly lower in subjects with APO E3/E3 (17.7%) than the subjects without APO E3/E3 genotype (41.9%; p = 0.01), and the corresponding odds ratio was 0.29 (95% confidence interval [CI], 0.11-0.78). In conclusion, this study provides strong evidence for the first time that APO E polymorphism is associated with the development of TBI-induced pituitary dysfunction. Present data demonstrated that APO E3/E3 genotype decreases the risk of hypopituitarism after TBI. The demonstration of the association between the APO E polymorphism and TBI may provide a new point of view in this field and promote further studies.


Retinal pigment epithelial damage, breakdown of the blood-retinal barrier, and retinal inflammation in dogs with primary glaucoma.
This paper aims to determine if abnormalities of the retinal pigment epithelium (RPE) and retinal inflammation occur in primary glaucoma. Twenty-three canine globes with primary glaucoma, goniodysgenesis, and elevated intraocular pressure were evaluated. Sections from 6 control and 23 glaucomatous canine globes were stained with hematoxylin and eosin, Griffonia simplicifolia isolectin B4, or immunohistochemically stained for CD3 or albumin. The retinal sections were evaluated with light microscopy for morphological and immunohistochemical evidence of pigmentary changes and inflammation. Abnormal pigmented cells including displaced RPE cells and macrophages (identified by lectin binding) were found in the neuroretinas and vitreous bodies of glaucomatous eyes. Other abnormalities included hypertrophy of RPE cells and loss of RPE continuity. Regions of neuroretina with more displaced pigment had fewer remaining neurons. Signs of retinal inflammation found in glaucomatous eyes included infiltration with leukocytes, retinal swelling, and albumin leakage from vessels. Accumulation of perivascular CD3-positive T lymphocytes also occurred in glaucomatous retinas. Chronic glaucomatous retinas had increased pigmentary changes, fewer neutrophils, and less swelling than acute glaucomatous retinas. Disruption of the RPE, increased permeability of the vascular endothelium, accumulation of inflammatory cells, and retinal swelling or thinning occur in canine primary glaucoma. The displacement of pigment and accumulation of inflammatory cells in the neuroretina suggests that inflammation may be an important contributor to retinal damage.


Further insights into the structure of the alternative oxidase: from plants to parasites.
The AOX (alternative oxidase) is a non-protonmotive ubiquinol-oxygen oxidoreductase that couples the oxidation of ubiquinol with the complete reduction of water. Although it has long been recognized that it is ubiquitous among the plant kingdom, it has only recently become apparent that it is also widely found in other organisms including some human parasites. In this paper, we review experimental studies that have contributed to our current understanding of its structure, with particular reference to the catalytic site. Furthermore, we propose a model for the ubiquinol-binding site which identifies a hydrophobic pocket, between helices II and III, leading from a proposed membrane-binding domain to the catalytic domain.


Rapid, Secure Drug Testing Using Fingerprint Development and Paper Spray Mass Spectrometry.
Paper spray mass spectrometry (PS-MS) is a technique that has recently emerged and has shown excellent analytical sensitivity to a number of drugs in blood. As an alternative to blood, fingerprints have been shown to provide a noninvasive and traceable sampling matrix. Our goal was to validate the use of fingerprint samples to detect cocaine use. Samples were collected on triangular pieces (168 mm2) of washed Whatman Grade I chromatography paper. Following application of internal standard, spray solvent and a voltage were applied to the paper before mass spectrometry detection. A fingerprint visualization step was incorporated into the analysis procedure by addition of silver nitrate solution and exposing the sample to ultraviolet light. Limits of detection for cocaine, benzoylecgonine, and methylecgonine were 1, 2, and 31 ng/mL respectively, with relative standard deviations < 33%. No matrix effects were observed. Analysis of 239 fingerprint samples yielded a 99% true-positive rate and a 2.5% false-positive rate, based on the detection of cocaine, benzoylecgonine, or methylecgonine with use of a single fingerprint. The method offers a qualitative and noninvasive screening test for cocaine use. The analysis method developed is rapid (4 min/sample) and requires no sample preparation.


Effect of DNA on the inactivation of O6-alkylguanine-DNA alkyltransferase by 9-substituted O6-benzylguanine derivatives.
Studies were carried out on the inactivation of pure human O6-alkylguanine-DNA alkyltransferase by 9-substituted O6-benzylguanine derivatives in the presence and absence of DNA. The addition of DNA increased the rate of inactivation of the alkyltransferase by O6-benzylguanine and its 9-methyl derivative but had little effect on the rate of inactivation by the 9-cyanomethyl derivative. In contrast, when O6-benzylguanine derivatives with larger 9-substituents such as ribose, 2'-deoxyribose, dihydrotestosterone, or 2-hydroxy-3-(isopropoxy)propyl were used, the addition of DNA was strongly inhibitory to the inactivation. In the case of O6-benzylguanine, O6-benzylguanosine, and O6-benzyl-2'-deoxyguanosine, these results were confirmed by directly measuring the rate of formation by the alkyltransferase of guanine, guanosine, or 2'-deoxyguanosine, respectively. The data indicated that the presence of DNA activated the alkyltransferase, rendering it more reactive with O6-benzylguanine or O6-benzyl-9-methylguanine, but that DNA interferes with the binding of inhibitors with larger 9-substituents, presumably by competing for the same binding site. Since these inactivators readily inactivate alkyltransferase in cells, the amount of cellular alkyltransferase bound to DNA must be small or readily exchangeable with the free form.


Paying the price? The impact of smoking and obesity on health inequalities in later life.
The aims of the study are twofold: (i) to explore the impact of socioeconomic position, as measured 13 years earlier, on cognitive functioning and mobility impairment in later life, and (ii) to explore the extent to which obesity and smoking status can explain socioeconomic inequalities in cognitive and mobility impairments in later life. Data from a nationally representative sample of Swedish adults aged 56-76 in 1991 who were re-interviewed 13 years later in 2004, was analysed to explore the impact of socioeconomic position, smoking, and obesity on cognitive and physical functioning in late life. The results showed that both smoking and obesity in late mid-life were stratified by socioeconomic position. Moreover, the results showed significant associations between socioeconomic position and both cognitive and physical functioning in later life. However, these inequalities in late life function could only partially be explained by the socioeconomic differences in smoking and obesity. The findings of this study suggest that socioeconomic differences in the rates of smoking and obesity may explain some, but not all, of the socioeconomic inequalities in physical and cognitive functioning during old age.


Intestinal Enterococcus abundance correlates inversely with excessive weight gain and increased plasma leptin in breastfed infants.
Epidemiological evidence indicates that breastfeeding provides protection against development of overweight/obesity. Nonetheless, a small subgroup of infants undergo excessive weight gain during exclusive breastfeeding, a phenomenon that remains unexplained. Breast milk contains both gut-seeding microbes and substrates for microbial growth in the gut of infants, and a large body of evidence suggests a role for gut microbes in host metabolism. Based on the recently established SKOT III cohort, we investigated the role of the infant gut microbiota in excessive infant weight gain during breastfeeding, including 30 exclusively breastfed infants, 13 of which exhibited excessive weight gain and 17 controls which exhibited normal weight gain during infancy. Infants undergoing excessive weight gain during breastfeeding had a reduced abundance of gut Enterococcus as compared with that observed in the controls. Within the complete cohort, Enterococcus abundance correlated inversely with age/gender-adjusted body-weight, body-mass index and waist circumference, body fat and levels of plasma leptin. The reduced abundance of Enterococcus in infants with excessive weight gain was coupled to a lower content of Enterococcus in breast milk samples of their mothers than seen for mothers in the control group. Together, this suggests that lack of breast milk-derived gut-seeding Enterococci may contribute to excessive weight gain in breastfed infants.


Proteinuria as an independent risk factor for contrast-induced acute kidney injury and mortality in patients with stroke undergoing cerebral angiography.
The correlation between proteinuria and contrast-induced acute kidney injury (CI-AKI) in patients with cerebrovascular disease is still unknown. To determine whether proteinuria is a risk factor for CI-AKI and death in patients with stroke undergoing cerebral angiography. Data from 2015 patients with stroke undergoing cerebral angiography between January 2009 and December 2013 were retrospectively collected. Clinical parameters were obtained from the hospital's computerized database. All variables were analyzed by univariate analysis and multivariate logistic regression analysis. CI-AKI was seen in 85 patients (4.2%). After adjustment for potential confounding risk factors, patients with proteinuria had a fivefold higher risk of CI-AKI than patients without proteinuria (OR=5.74; 95% CI 2.23 to 14.83; p<0.001). Other independent risk factors for CI-AKI were estimated glomerular filtration rate <60 mL/min/1.73 m2, anemia, and a high National Institute of Health Stroke Scale score. Proteinuria did not increase in-hospital mortality (OR=1.25; 95% CI 0.49 to 3.17; p=0.639) but did increase 1-year mortality (HR=2.30, 95% CI 1.55 to 3.41, p<0.001). Proteinuria is an independent risk factor for CI-AKI and 1-year mortality in patients with stroke undergoing cerebral angiography. More attention should be paid to the development of CI-AKI in patients with stroke with proteinuria.


van Leeuwenhoek microscopes-where are they now?
When Antonie van Leeuwenhoek died, he left over 500 simple microscopes, aalkijkers (an adaption of his microscope to allow the examination of blood circulation in the tails of small eels) and lenses, yet now there are only 10 microscopes with a claim to being authentic, one possible aalkijker and six lenses. He made microscopes with more than one lens, and possibly three forms of the aalkijker. This paper attempts to establish exactly what he left and trace the fate of some of the others using the earliest possible documents and publications.


Calcium-sensitive pyruvate dehydrogenase phosphatase is required for energy metabolism, growth, differentiation, and infectivity of Trypanosoma cruzi.
In vertebrate cells, mitochondrial Ca2+ uptake by the mitochondrial calcium uniporter (MCU) leads to Ca2+-mediated stimulation of an intramitochondrial pyruvate dehydrogenase phosphatase (PDP). This enzyme dephosphorylates serine residues in the E1α subunit of pyruvate dehydrogenase (PDH), thereby activating PDH and resulting in increased ATP production. Although a phosphorylation/dephosphorylation cycle for the E1α subunit of PDH from nonvertebrate organisms has been described, the Ca2+-mediated PDP activation has not been studied. In this work, we investigated the Ca2+ sensitivity of two recombinant PDPs from the protozoan human parasites Trypanosoma cruzi (TcPDP) and T. brucei (TbPDP) and generated a TcPDP-KO cell line to establish TcPDP's role in cell bioenergetics and survival. Moreover, the mitochondrial localization of the TcPDP was studied by CRISPR/Cas9-mediated endogenous tagging. Our results indicate that TcPDP and TbPDP both are Ca2+-sensitive phosphatases. Of note, TcPDP-KO epimastigotes exhibited increased levels of phosphorylated TcPDH, slower growth and lower oxygen consumption rates than control cells, an increased AMP/ATP ratio and autophagy under starvation conditions, and reduced differentiation into infective metacyclic forms. Furthermore, TcPDP-KO trypomastigotes were impaired in infecting cultured host cells. We conclude that TcPDP is a Ca2+-stimulated mitochondrial phosphatase that dephosphorylates TcPDH and is required for normal growth, differentiation, infectivity, and energy metabolism in T. cruzi Our results support the view that one of the main roles of the MCU is linked to the regulation of intramitochondrial dehydrogenases.


Clinical spectrum of radiation induced pericarditis.
A series of seven patients diagnosed as having radiation induced pericarditis were reviewed and compared to those reported in the literature. They constituted 6.8 per cent of all patients with the diagnosis of pericarditis seen in a five-year period. These patients had the diagnosis proven by autopsy or by the absence of tumor cells from the pericardial fluid and/or surgical biopsy. The echocardiogram and cardiac catheterization were found to be specific diagnostic tools. Recommendations are made for a therapeutic approach to these patients. The diagnosis of this condition depends on a high index of suspicion, and close follow-up of patients who have received intense radiotherapy to the chest. Recommendations are made for prophylaxis. We challenge the pessimistic approach towards the cancer patient who develops recurrent pericardial fluid or pleural effusion due to radiation induced pericarditis. We advocate an aggressive diagnostic approach and therapy which should be related to the degree of hemodynamic impairment.


Central corneal thickness in primary angle closure and open angle glaucoma.
To quantitatively analyze central corneal thickness (CCT) in patients with primary angle closure glaucoma (PACG) and primary open-angle glaucoma (POAG), and to evaluate its correlation with severity of glaucoma. In this retrospective study, records of patients with previously diagnosed POAG or PACG at a tertiary glaucoma service were reviewed. CCT was measured by ultrasound pachymetry. Mean deviation (MD) on visual field (VF) testing was recorded for glaucoma severity determination. CCT and age- and sex-adjusted CCT were compared among the study groups using Student's t-test and analysis of covariance (ANCOVA), respectively. Univariate and multivariate regression models were used for correlation of age, MD and CCT. A total of 115 patients with PACG, 215 with POAG, and 100 normal controls were included with mean age of 64.1 ± 10.4, 59.9 ± 10.5, and 62.04 ± 10.80 years, respectively. CCT was thicker in PACG eyes (545.5 ± 46.1 μm) as compared to POAG eyes (531.7 ± 37.3 μm) and controls (531.0 ± 38.3 μm) even after age and gender adjustment (ANCOVA, P = 0.05). CCT was found to decrease with increasing age only in the POAG group (β = -0.57, P = 0.01). Disease severity (MD of VF) was significantly and inversely correlated with CCT in both POAG and PACG eyes (β = 1.89, P = 0.02; and β = 1.38, P = 0.04, respectively) after age and sex correction. PACG eyes had thicker CCT as compared to POAG and normal healthy eyes in Iranian subjects. Severity of the disease was inversely correlated with CCT in eyes with both POAG and PACG.


Iron deficiency leads to inhibition of oxygen transfer and enhanced formation of virulence factors in cultures of Pseudomonas aeruginosa PAO1.
Pseudomonas aeruginosa PAO1 was recently found to exhibit two remarkable physiological responses to oxidative stress: (1) a strong reduction in the efficiency of oxygen transfer from the gas phase into the liquid phase, thus causing oxygen limitation in the culture and (2) formation of a clear polysaccharide capsule on the cell surface. In this work, it has been shown that the iron concentration in the culture plays a crucial role in evoking these phenomena. The physiological responses of two P. aeruginosa PAO1 isolates (NCCB 2452 and ATCC 15692) were examined in growth media with varied iron concentrations. In a computer-controlled bioreactor cultivation system for controlled dissolved oxygen tension (pO2), a strong correlation between the exhaustion of iron and the onset of oxygen limitation was observed. The oxygen transfer rate of the culture, characterized by the volumetric oxygen transfer coefficient, kLa, significantly decreased under iron-limited conditions. The formation of alginate and capsule was more strongly affected by iron concentration than by oxygen concentration. The reduction of the oxygen transfer rate and the subsequent oxygen limitation triggered by iron deficiency may represent a new and efficient way for P. aeruginosa PAO1 to adapt to growth conditions of iron limitation. Furthermore, the secretion of proteins into the culture medium was strongly enhanced by iron limitation. The formation of the virulence factor elastase and the iron chelators pyoverdine and pyochelin also significantly increased under iron-limited conditions. These results have implications for lung infection of cystic fibrosis patients by P. aeruginosa in view of the prevalence of iron limitation at the site of infection and the respiratory failure leading to death.