An optimal experimental design for the development of preservative heart solutions.
The aim of this study was to determine the optimal composition of a new medium for long-term hypothermic heart preservation. The independent effects of 19 compounds were evaluated using an in vitro porcine model. Tissue viability was assessed by measuring the reduction of methyltetrazolium salt, oxygen consumption and energetic compound levels on myocardial biopsies after 24-, 48- or 72-hour incubation periods. Screening of several compounds at two concentrations was performed to greatly reduce the number of experiments. Pyruvate, aspartic acid, chlorpromazine and polyethylene glycol displayed protective properties, whereas calcium (2 mmol/liter), nifedipine, mannitol, magnesium (16 mmol/liter) and reduced glutathione showed deleterious effects. On the basis of these data, the composition of a new preservation solution (Group 1, n = 6) was compared with St Thomas solution (Group II, n = 6) in an isolated, 24-hour pig heart preservation model. During reperfusion, left ventricular developed pressure and coronary blood flow were significantly higher (p <.01) in Group I, suggesting better preservation. Our technique allows for rapid and efficient screening of many compounds currently used in the composition of preservation solutions for cardiac surgery.


Isolation and characterization of a novel BcMF14 gene from Brassica campestris ssp. chinensis.
A putative RALF (rapid alkalinization factor)-like gene (GenBank accession number EF523517), named BcMF14, was isolated from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis) by rapid amplification of cDNA ends (RACE) based on a cDNA-AFLP differential fragment exclusively expressed in fertile line. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) discovered that BcMF14 was prominently expressed in stage four and five flower buds of fertile line, no expression in vegetative structures or in sterility line. Detailed RT-PCR illuminated its strong expression in stamens. Successful suppression of BcMF14 gene expression greatly reduced the normal pollen grains. The frequency of abnormal pollen grains was 48.95% in the mutant with many shriveled pollen grains with irregular shape and some larger ones with deep hollows along the germination ditch. Pollen germination was stopped because of the severely twisted pollen tubes. These results demonstrate a potential role of the BcMF14 gene in the development of male gametogenesis in Chinese cabbage.


Differential effects of atorvastatin treatment and withdrawal on pentylenetetrazol-induced seizures.
Statins are selective inhibitors of 3-hydroxyl-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme of the mevalonate pathway for cholesterol biosynthesis. Increasing evidence indicates that statins, particularly atorvastatin, are neuroprotective in several conditions, including stroke, cerebral ischemia, traumatic brain injury, and excitotoxic amino acid exposure. However, only a few studies have investigated whether statins modulate seizure activity. In the current study we investigated whether atorvastatin or simvastatin alters the seizures induced by pentylenetetrazol (PTZ), a classical convulsant. Adult male Wistar rats were treated with atorvastatin or simvastatin for 7 days (10 mg/kg/day). Seizure activity was induced by PTZ (60 mg/kg, i.p.), and evaluated by behavioral and electrographic methods. Cholesterol levels were determined by a standard spectrophotometric method. Blood-brain barrier (BBB) permeability was assessed by the fluorescein method. Atorvastatin levels in the plasma and cerebral cortex were determined by high-performance liquid chromatography tandem mass spectrometry. We found that oral atorvastatin treatment increased the latency to PTZ-induced generalized seizures. In contrast, when the 7-day atorvastatin treatment was withheld for 1 day (i.e., atorvastatin withdrawal), PTZ-induced seizures were facilitated, as evidenced by a decrease in the latency to clonic and generalized tonic-clonic seizures induced by PTZ. In contrast, simvastatin treatment for 7 days (10 mg/kg/day, p.o.), with or without withdrawal, did not alter PTZ-induced seizures. Interestingly, the effects of atorvastatin treatment and withdrawal were not accompanied by changes in plasma or cerebral cortex cholesterol levels or in the BBB permeability. Atorvastatin levels in the plasma and cerebral cortex after 7 days of treatment were above the half maximal inhibitory concentration for inhibition of HMG-CoA reductase, whereas atorvastatin was not detectable in the plasma or cerebral cortex following a 24 h washout period (atorvastatin withdrawal). We conclude that atorvastatin treatment and withdrawal have differential effects on pentylenetetrazol-induced seizures, which are not related to changes in plasma or cerebral cortex cholesterol levels or in BBB permeability. Additional studies are necessary to evaluate the molecular mechanisms underlying our findings as well as its clinical implications.


The detection of African trypanosomes in goats reared in tsetse infested villages of Eastern Zambia.
Control programmes for African animal trypanosomiasis (AAT) in livestock have been mainly focused on cattle with very little focus on goats, an important reservoir for the disease. Using the polymerase chain reaction (PCR), this study investigated trypanosome infection in village goats in Mambwe, a rural District in Eastern Zambia. Filter paper blood spots were collected from 326 goats and tested for infection with Trypanosoma congolense, Trypanosoma vivax and Trypanosoma brucei s.l. using ribosomal RNA internal transcribed spacers (ITS)-PCR. The frequency of trypanosomes from the sampled goats was 4.6% (95% CI = 2.3-6.8). Results indicated significantly high infections with Trypanosoma vivax (4.0%; 95% CI = 1.9-6.1) than T. congolense (0.6%; 95% CI = - 0.2 to 1.5), and T. brucei (0.0%), P = 0.04. Findings show the circulation of trypanosomes that causes AAT in goats and that they may pose serious threats to not only goats but also to other livestock reared alongside goats.


Protein carboxyl-O-methyltransferase activity in cultured C-1300 neuroblastoma cells.
Protein carboxylmethyltransferase (PCM) has been identified in a variety of tissues derived from neural crest anlage, including in vivo C-1300 murine neuroblastoma (MNB). These observations have stimulated interest in further defining the role of PCM as a potential modulator of neoplastic cell behavior. The subcellular distribution and kinetic behavior of PCM have been characterized in a tissue culture line derived from the C-1300 murine neuroblastoma (clone NB41A3). The specific and total activities of PCM in the presence and absence of exogenous substrate were determined in subcellular fractions of MNB cells prepared by differential centrifugation. In the presence of exogenous substrate (+ gelatin), 40% of the total PCM activity was present in the 100,000 g supernatant fraction and 41% in the 800 g particulate fraction, whereas the higher specific activity of PCM was present in the 100,000 g supernatant fraction. Enzyme activity measured in the absence of gelatin, which reflects the concentration of endogenous methyl acceptor proteins in a cell fraction, was negligible. This activity represented less than 1.6 and 0.4% of the total PCM activity present in the 800 g particulate and 100,000 g soluble fractions respectively. Cytosolic PCM had an apparent Km of 13.9 x 10(-6) M for AdoMet and a Vmax of 33 pmoles per min per mg protein. Cytoplasmic PCM was inhibited competitively by S-adenosylhomocysteine (Ki equal 0.2 microM). These data demonstrate that the specific activity of PCM was greatest in the soluble component of subcellular fractions prepared from cultured MNB cells. This distribution pattern of PCM is similar to that observed in the C-1300 MNB tumor grown in situ and in non-malignant tissues. In contrast to the latter tissues, cultured MNB cells exhibited low PCM activity when assayed in the absence of exogenous substrate.


Transport protein evolution deduced from analysis of sequence, topology and structure.
The vast majority of well studied transmembrane channels, secondary carriers, primary active transporters and group translocators are believed to have arisen vis intragenic duplication events from simple channel-forming peptides with just 1-3 transmembrane α-helical segments, found ubiquitously in nature. Only a few established channel-forming proteins appear to have evolved via other pathways. The proposed pathway for the evolutionary appearance of the five types of transport proteins involved intragenic duplication of transmembrane pore-forming peptide-encoding genes, giving rise to channel proteins. These gave rise to single protein secondary carriers which upon superimposition of addition protein domains and proteins, including energy-coupling proteins and extracytoplasmic receptors, gave rise to multidomain, multicomponent carriers, primary active transporters and group translocators. Some of the largest and best characterized superfamilies of these transmembrane transport proteins are discussed from topological and evolutionary standpoints.


Studying the structural and folding features of long-sequence trichobrachin peptides.
In this theoretical study, the folding processes of long-sequence trichobrachin peptides (i.e., TB IIb peptides) were investigated by molecular dynamics methods. The formation of various helical structures (i.e., 310 -, α-, and left-handed α-helices) was studied with regard to the entire sequence of peptides, as well as to each amino acid. The results pointed out that TB IIb molecules showed a propensity to form helical conformations, and they could be characterized by 310 -helical structure rather than by α-helical structure. The formation of local (i.e., i←i+3 and i←i+4) as well as of non-local (i.e., i←i+n, where n>4; and all i→i+n) H-bonds was also examined. The results revealed that the occurrence of local, helix-stabilizing H-bonds was in agreement with the appearance of helical conformations, and the non-local H-bonds did not produce relevant effects on the evolution of helical structures. Based on the data obtained by our structural investigation, differences were observed between the TB IIb peptides, according to the type of amino acid located in the 17th position of their sequences. In summary, the folding processes were explored for TB IIb molecules, and our theoretical study led to the conclusion that these long-sequence peptaibols showed characteristic structural and folding features.


Phosphorylation of synapsin I and dynamin in rat forebrain synaptosomes: modulation by sigma (sigma) ligands.
The effects of sigma (sigma) ligands on protein phosphorylation were examined in crude, rat forebrain synaptosomes. Synaptosomes were prelabelled with 32P(i) and incubated with the sigma ligands 1,3-di-o-tolylguanidine (DTG), (+)pentazocine and (-)pentazocine (3, 10, 30, 100, 300 microM), or haloperidol, reduced haloperidol, and (+)SKF 10,047 (100 microM). Aliquots were then incubated for 10 s in control (5 mM K+) or depolarising buffer (41 mM K+). All the sigma ligands increased basal phosphorylation of synapsin Ib and other proteins including dynamin, and inhibited the depolarisation-dependent increase in phosphorylation of synapsin Ib in synaptosomes. The effects of these ligands are not directly on protein kinases or protein phosphatases. This indicates that the sigma ligands are mediating their effects via interaction with sigma binding sites, and suggest, for the first time, that protein phosphorylation may be one mechanism through which sigma ligands produce their biological effects.


Response tests of a LaCl(3):Ce scintillation detector with low energy prompt gamma rays from boron and cadmium.
The yield of 478 and 558 keV gamma-rays have been measured from water samples containing 0.031-0.500 wt. % boron and 0.0625-0.500 wt. % cadmium, respectively, using a cylindrical 76 mm × 76 mm (height × diameter) LaCl(3):Ce detector. Inspite of interferences between detector-associated and the sample-associated prompt gamma rays, the LaCl(3):Ce detector has excellent resolution for the low energy prompt gamma-rays. An excellent agreement has been observed between the experimental and calculated yield of boron and cadmium prompt gamma ray from water samples.


Ecological comparisons of robber fly species (Diptera: Asilidae) coexisting in a neotropical forest.
Data regarding seasonal abundance, microhabitat preference, and diet were collected over 3 field seasons for adults of 15 robber fly species on Barro Colorado Island, Panama. The species comprised 2 distinct "thermal guilds"; light-seeking (hereafter LS) species foraged in sunlit areas, while shade-seeking (SS) species foraged only in deep shade. All species were rare during the dry season. During the rainy months, most SS species had flight periods of 2-3 months, and no temporal segregation was apparent. In contrast, most LS species had flight periods of only 4-6 weeks, and a distinct sequence of occurrence was evident during 2 years of censusing. Most SS and LS species displayed a pronounced specificity for perches of a particular substrate type. However, the ranges of perching heights utilized varied considerably among species. Dietary comparisons revealed that mean and maximum prey sizes increased with increasing robber fly size, while minimum prey sizes were constant. Robber fly species <20 mg fed primarily upon nematocerous Diptera, whereas larger species generally fed upon a wide variety of prey types. For each thermal guild, the actual mean overlap for a particular niche dimension was compared to mean overlaps generated by randomly assigning species to thermal guilds. No significant differences from the random null hypothesis were found for the SS guild. However, niche complementarity between dietary and spatial overlaps and dietary overlap was apparent among the 5 large LS species.


RNA m6A methylation promotes the formation of vasculogenic mimicry in hepatocellular carcinoma via Hippo pathway.
Vasculogenic mimicry (VM) formed by aggressive tumor cells to mimic vasculogenic networks plays an important role in the tumor malignancy of HCC. However, the pathogenesis underlying VM is complex and has not been fully defined. m6A is a common mRNA modification and has many biological effects. However, the relationship between m6A and VM remains unclear. In this research, we found that m6A methyltransferase METTL3 in HCC tissues was positively correlated with VM. The m6A level of mRNA significantly increased in 3D cultured cells treated with VEGFa and was related to VM formation. Transcriptome sequencing analysis of 3D cultured cells with knockdown Mettl3 showed that the Hippo pathway was involved in m6A-mediated VM formation. Further mechanism research indicated that the m6A modification of YAP1 mRNA affected the translation of YAP1 mRNA. In conclusion, m6A methylation plays a key role in VM formation in HCC. METTL3 and YAP1 could be potential therapeutic targets via impairing VM formation in anti-metastatic strategies.


Percutaneous management of a traumatic arteriovenous fistula.
The diagnosis of arteriovenous fistulae can be inhibited by inadequate clinical information and the presence of non specific signs. We describe how a patient was diagnosed with an arteriovenous fistula 19 years following penetrating trauma. We describe our successful treatment of this fistula using radiological endovascular techniques.


The impact of depression and diabetes mellitus on older workers' functioning.
Using the International Classification of Functioning, Disability and Health as a framework, this study investigates the impact of depression and diabetes mellitus on older workers' functioning (problems with concentration, physical functioning, need for recovery and work and social participation restrictions). The study focuses on how these chronic conditions, in their interaction with the work context, affect older workers' functioning, which may be an important precursor of early retirement. Older workers (≥ 45years) with depression (n=127) or diabetes mellitus (n=107) enrolled in the prospective Maastricht Cohort Study (MCS) were followed between October 2008 and October 2012. Linear, logistic and Cox regression analyses were performed to investigate the effect of these health conditions on workers' functioning compared to a reference group of older workers without a chronic condition (n=1612). The interaction with participants' working conditions (psychological job demands, decision latitude and strenuous work) was also analysed. Compared to the reference group, depression and diabetes mellitus were (over time) positively related with need for recovery caseness and restrictions in social participation but not with restrictions in work participation. Depression was positively related with concentration problems and need for recovery, whereas diabetes mellitus was negatively related with physical functioning. Finally, the relationship between functioning and depression and diabetes mellitus depends on working conditions. Older workers with depression or diabetes mellitus are vulnerable to losses in specific domains of functioning. The impact on functioning varies across working conditions, providing insight for disease-tailored preventive measures.


Effects of high-fat diet on sympathetic neurotransmission in mesenteric arteries from Dahl salt-sensitive rat.
Obesity hypertension is driven by sympathetic neurotransmission to the heart and blood vessels. We tested the hypothesis that high-fat diet (HFD)-induced hypertension is driven by sympathetic neurotransmission to mesenteric arteries (MA) in male but not female Dahl salt-sensitive (Dahl ss) rat. Rats were fed a control diet (CD; 10 kcal% from fat) or HFD (60 kcal% from fat) beginning at 3 weeks (wk) of age; measurements were made at 10-, 17- and 24-wk. Body weight increased with HFD, age and sex. Mean arterial pressure (MAP) was higher in HFD versus CD rats from both sexes at 17- and 24-wk. MA constriction measured using pressure myography, and electrical field stimulation (EFS, 0.2-30 Hz) was greater in HFD versus CD in males at 17-wk; this was not due to changes in α2 autoreceptor or norepinephrine transporter (NET) function. Prazosin (α1-AR antagonist) and suramin (P2 receptor antagonist) inhibited neurogenic MA constriction equally in all groups. Arterial reactivity to exogenous norepinephrine (NE; 10-8 - 10-5 M) was lower in HFD versus CD at 10-wk in males. Female MA reactivity to exogenous ATP was lower at 24-weeks compared to earlier time points. HFD did not affect tyrosine hydroxylase (TH) or the vesicular nucleotide transporter (VNUT) nerve density in MA from both sexes. NE content was lower in MA but higher in plasma at 24-wk compared to 10- and 17-wk in both sexes. In conclusion, HFD-induced hypertension is not driven by increased sympathetic neurotransmission to MA in male and female Dahl ss rats.


Differential regulation of phosphoinositide and phosphatidylcholine hydrolysis by protein kinase C-beta 1 overexpression. Effects on stimulation by alpha-thrombin, guanosine 5'-O-(thiotriphosphate), and calcium.
Rat 6 fibroblasts that stably overexpress cDNA for the beta 1 isozyme of protein kinase C (PKC3 cells) were used to determine the effect of protein kinase C (PKC) overexpression on hormonal stimulation of phospholipid hydrolysis. In control Rat 6 cells, inositol trisphosphate levels (InsP3) were increased 9-fold in 15 s in response to 10 nM alpha-thrombin, compared with only a 2-fold increase in PKC3 cells. PKC overexpression also inhibited thrombin-stimulated production of 1,2-diacylglycerol, the other product of phosphatidylinositol 4,5-bisphosphate hydrolysis, by 73% at 15 s. In permeabilized cells, PKC overexpression greatly reduced guanosine thiotriphosphate-stimulated InsP3 accumulation, but did not affect InsP3 stimulation by increased free calcium concentration. These data suggest that desensitization of thrombin-stimulated phosphoinositide-phospholipase C is enhanced by PKC-beta 1 overexpression and may involve modulation of G-protein/phospholipase C coupling. In contrast, thrombin was 4.5-fold more effective in stimulation of phosphatidylcholine-phospholipase D activity in PKC3 cells than in control cells, as determined by phosphatidylethanol formation. In permeabilized cells, guanosine thiotriphosphate also stimulated phospholipase D activity more effectively in PKC3 cells than in control cells, suggesting that upregulation of phospholipase D activity by PKC overexpression occurs distal to the thrombin receptor. These results suggest that PKC may act as a switch to up-regulate phosphatidylcholine-phospholipase D and down-regulate phosphoinositide-phospholipase C stimulations.


Gene delivery methods and genome editing of human pluripotent stem cells.
Induced pluripotent stem cells derived from normal somatic cells could be utilized to study tumorigenesis through overexpression of specific oncogenes, downregulation of tumor suppressors and dysregulation of other factors thought to promote tumorigenesis. Therefore, effective approaches that provide direct modifications of induced pluripotent stem cell genome are extremely needed. Emerging strategies are expected to provide the ability to more effectively introduce diverse genetic alterations, from as small as single-nucleotide modifications to whole gene amplification or deletion, all with a high degree of target specificity. To date, several techniques have been applied in stem cell studies to directly edit cell genome (ZFNs, TALENs or CRISPR/Cas9). In this review, we summarize specific gene delivery strategies that were applied to stem cell studies together with genome editing techniques, which enable a direct modification of endogenous DNA sequences in the context of cancer studies.


Canadian senate committee to review drug laws and policies.
Canada's Senate voted on 11 April 2000 to establish a Special Committee to conduct a comprehensive review of Canadian drug laws and policies.


Neoadjuvant Immunotherapy in Early, Triple-Negative Breast Cancers: Catching Up with the Rest.
Despite breast cancer being long thought to be "immunologically cold," within early, triple-negative breast cancer (TNBC), there has been exciting advances with the use of immune checkpoint modulation combined with neoadjuvant chemotherapy. We review the major trials that have investigated combination immunochemotherapy in the neoadjuvant setting, reviewing both the pathological complete response rates and the maturing data regarding event-free and overall survival. Strategies to deescalate adjuvant therapy in patients with preserving excellent clinical outcome, as well as exploration of combinatorial adjuvant therapies to improve outcome in those with extensive residual are the next-generation challenges. In addition to refinement of existing biomarkers, such as PD-L1, TILs, and tumor mutational burden (TMB), exploration of topics like the microbiome as both a biomarker and a therapeutic has shown promise in other cancer types, which motivates investigating these in breast cancer.


A new SVWC protein functions as a pattern recognition protein in antibacterial responses in Chinese mitten crab (Eriocheirsinensis).
Because invertebrates lack acquired immunity, they rely primarily on the innate immune system to defend themselves against viral and bacterial infections. SVWC, also called Vago, is a class of small-molecule proteins characterized by a single von Willebrand factor C-domain and appears to be restricted to arthropods. It has been reported that SVWC is involved in antiviral immunity in invertebrates, but whether it is involved in antimicrobial immunity and the mechanism of its involvement in antimicrobial immunity remains unclear. In this study, we identified a novel SVWC gene in Eriocheir sinensis and named it EsSVWC. EsSVWC was found to respond positively to bacterial stimulation and to regulate the expression of related antimicrobial peptides (AMPs). The EsSVWC protein recognized and bound to a variety of pathogen-associated molecular patterns (PAMPs) but did not exhibit direct bactericidal effects. Thus, the EsSVWC protein in crabs helps resist bacterial infection and improve survival rates. In summary, EsSVWC may regulate the innate immune system of crabs in response to microbial invasion in an indirect manner.


Psychological and sexual functioning in women with vulvar vestibulitis.
The purpose of the study was to compare psychological and sexual functioning in women with vestibulitis with healthy controls. It was hypothesized that women with vestibulitis would experience greater psychological stress and sexual dissatisfaction than controls. Fifty-two women with vestibulitis recruited from a vulvovaginal disease clinic and 46 healthy controls recruited from an outpatient gynecology clinic completed five standardized measures of psychological and sexual functioning. Multivariate analyses of variance and covariance were used to examine group differences. Women with vestibulitis reported significantly higher scores than controls on the measures of depression (p < or = 0.001), psychological distress (p < or = 0.001) and sexual depression (p < or = 0.001). They reported significantly lower scores on the measures of sexual satisfaction (p < or = 0.001), sexual behavior (p < or = 0.001) and sexual self-esteem (p < or = 0.01). The results of this study highlight the importance of addressing psychological distress and sexual dissatisfaction in women with vestibulitis.


Seasonal dimorphism of foliage in Californian coastal sage scrub.
A pattern of seasonal variation in leaf size and position relative to the stem is reported for twelve mesophyllous shrubs in the coastal sage scrub of Mediterranean-climate areas of southern California. Leaves of reduced size, produced on axillary short shoots late in the growing season, persist through the dry season while many of the larger leaves on main stems are abscissed. The short-shoot leaves of Salivia mellifera have the same density thickness as those of the main stem, leaving open the question of whether the smaller summer leaves have physiological mechanisms for drought tolerance beyond a reduction in total transpiring surface. In Californian coastal shrublands seasonal dimorphism of leaves appears limited to mesophyllous shrubs. It is also found in analogues in comparable climatic regions of the Mediterranean, Chile and South Africa.


Photochemical Functionalization of Helicenes.
Herein, a visible-light photochemical approach for practical helicene functionalization at very mild reaction conditions is described. The photochemical reactions allow for the regiospecific and innate late-stage functionalization of helicenes and are easily executed either through the activation of C(sp2 )-Br bonds in helicenes using K2 CO3 as inorganic base or direct C(sp2 )-H helicene bond functionalization under oxidative photoredox reaction conditions. Overall, using these transformations six different functional groups are introduced to the helicene scaffold through C-C and four different C-heteroatom bond-forming reactions.


Disordered regions endow structural flexibility to shell proteins and function towards shell-enzyme interactions in 1,2-propanediol utilization microcompartment.
Intrinsically disordered regions in proteins have been functionally linked to the protein-protein interactions and genesis of several membraneless organelles. Depending on their residual makeup, hydrophobicity or charge distribution they may remain in extended form or may assume certain conformations upon biding to a partner protein or peptide. The present work investigates the distribution and potential roles of disordered regions in the integral proteins of 1,2-propanediol utilization microcompartments. We use bioinformatics tools to identify the probable disordered regions in the shell proteins and enzyme of the 1,2-propanediol utilization microcompartment. Using a combination of computational modelling and biochemical techniques we elucidate the role of disordered terminal regions of a major shell protein and enzyme. Our findings throw light on the importance of disordered regions in the self-assembly, providing flexibility to shell protein and mediating its interaction with a native enzyme.Communicated by Ramaswamy H. Sarma.


Retraction: Cognitive-behavioural therapy for personal recovery of patients with schizophrenia: a systematic review and meta-analysis.
[This retracts the article DOI: 10.1136/gpsych-2018-100040.].


Effects of pimozide on the response of smooth muscle to non-dopamine agonists and calcium.
Pimozide inhibits acetylcholine and histamine-induced contractions of the isolated guinea-pig ileum and noradrenaline-induced contractions of the isolated rat vas deferens in a non-competitive manner. In order to explore the nature of this inhibitory effect, we investigated pimozide's effects on calcium-induced contractions of deplorized guinea-pig ileum. In this case the antagonism elicited was competitive. Therefore caution is indicated in the use of pimozide to provide indirect proof of the existence of peripheral dopamine receptors.


Nudge strategies for behavior-based prevention and control of neglected tropical diseases: A scoping review and ethical assessment.
Nudging, a strategy that uses subtle stimuli to direct people's behavior, has recently been included as an effective and low-cost behavior change strategy in low- and middle- income countries (LMIC), targeting behavior-based prevention and control of neglected tropical diseases (NTDs). The present scoping review aims to provide a timely overview of how nudge interventions have been applied within this field. In addition, the review proposes a framework for the ethical consideration of nudges for NTD prevention and control, or more broadly global health promotion. A comprehensive search was performed in several databases: MEDLINE, PsycINFO, and Embase (Ovid), Web of Science Core Collection, CINAHL, ERIC and Econ.Lit (EBSCO), as well as registered trials and reviews in CENTRAL and PROSPERO to identify ongoing or unpublished studies. Additionally, studies were included through a handpicked search on websites of governmental nudge units and global health or development organizations. This scoping review identified 33 relevant studies, with only two studies targeting NTDs in particular, resulting in a total of 67 nudge strategies. Most nudges targeted handwashing behavior and were focused on general health practices rather than targeting a specific disease. The most common nudge strategies were those targeting decision assistance, such as facilitating commitment and reminder actions. The majority of nudges were of moderate to high ethical standards, with the highest standards being those that had the most immediate and significant health benefits, and those implemented by agents in a trust relationship with the target audience. Three key recommendations should inform research investigating nudge strategies in global health promotion in general. Firstly, future efforts should investigate the different opportunities that nudges present for targeting NTDs in particular, rather than relying solely on integrated health promotion approaches. Secondly, to apply robust study designs including rigorous process and impact evaluation which allow for a better understanding of 'what works' and 'how it works'. Finally, to consider the ethical implications of implementing nudge strategies, specifically in LMIC.


Multivariate statistics and hydrogeochemistry of deep groundwater at southwestern part of Bangladesh.
Multivariate statistics and GIS alone with geochemical modeling were applied to investigate the hydro-geochemical characteristics of groundwater and their spatial distribution in the deep aquifer system of Bagherpara Upazila, Bangladesh. This Upazila consists of an area of 308.3 km2 and local people mostly rely on groundwater to meet the drinking water requirements. Water samples from one hundred randomly selected deep tube wells (152-198 m) were collected and analyzed for 14 water quality parameters to characterize the hydro-geochemical properties. The groundwater shows slight alkaline in nature throughout the study area. Most of the water samples were turbid and 68% of them failed to meet the drinking water quality standard prescribed by the World Health Organization (WHO). TDS concentration ranges between 280 mg/L and 1040 mg/L, with a mean value of 446.20 (±122.19) mg/L. The general order of cation and anion along the study area were Ca2+>Mg2+>Na+>K+ and HCO3 ->Cl->PO4 3->SO4 2->NO3 - respectively. Carbonate weathering is the dominant process for releasing ions in groundwater. Besides, the ion exchange process is active in the study area, which leads to the reduction of Na+ ions. Gibb's plot suggests a rock dominance inheritance controls the dissolution and precipitation of minerals along with the ion-exchange process, and ultimately dictates the groundwater chemistry. Besides, the Piper diagram showed that Ca2+-Mg2+-SO4 2- is the dominant water type in 65% of the samples followed by Ca2+-Mg2+-HCO3 - water type (35%). The abundance of Ca2+ and Mg2+ ions and the alkaline nature of groundwater indicate mixed geochemical facies and reverse exchange reactions. The principal component analysis (PCA) reveals that weathering and leaching of host rocks was the main natural source, while municipal solid waste dumping, sewage discharge, and fertilizer application could be other anthropogenic factors that affect the groundwater geochemistry. In fine, the chemical characteristics of groundwater were acquired through rock dissolution, percolation, and reverse exchange process.


Detection of antigen specific rosette formation with the FACS.
Donors previously sensitized to conventional antigens PPD and KLH were evaluated for their antigen binding responses, utilizing a rosette forming technique with antigen-conjugated autologous erythrocytes. Reactivity is directly correlated with prior sensitization. Furthermore, antigen specificity is suggested by inhibition of rosette formation by prior incubation with the relevant antigen. The frequency of RFCs detected cytofluorometrically was compared with conventional fluorescent microscopy determinations. RFCs detected in this manner were identified as antibody armed monocytes by cell depletion and histochemical studies. The usefulness of the rosette forming technique for the routine evaluation of donor immunity is discussed.


AIT (allergen immunotherapy): a model for the "precision medicine".
The interpretation of medical approaches, especially therapy, evolved rapidly in the last century. Starting from the simple description of symptoms, we moved to the pathophysiological descriptions, to the evidence-based medicine, until the so-called "precision medicine". This latter can be defined as a structural model aimed at customizing healthcare, with medical decisions/products tailored on an individual patient at a highly detailed level. In this sense, allergen immunotherapy represents an optimal model of "precision medicine", since we know and describe symptoms, function, aetiological agents at molecular level, and we have the possibility to intervene on the natural history of the disease. If considered under the point of view of pharmaco-economy, that is prescribing the optimal treatment to the right patient, allergen immunotherapy represents an almost-ideal model of precision medicine.


Solution structure of the Big domain from Streptococcus pneumoniae reveals a novel Ca2+-binding module.
Streptococcus pneumoniae is a pathogen causing acute respiratory infection, otitis media and some other severe diseases in human. In this study, the solution structure of a bacterial immunoglobulin-like (Big) domain from a putative S. pneumoniae surface protein SP0498 was determined by NMR spectroscopy. SP0498 Big domain adopts an eight-β-strand barrel-like fold, which is different in some aspects from the two-sheet sandwich-like fold of the canonical Ig-like domains. Intriguingly, we identified that the SP0498 Big domain was a Ca(2+) binding domain. The structure of the Big domain is different from those of the well known Ca(2+) binding domains, therefore revealing a novel Ca(2+)-binding module. Furthermore, we identified the critical residues responsible for the binding to Ca(2+). We are the first to report the interactions between the Big domain and Ca(2+) in terms of structure, suggesting an important role of the Big domain in many essential calcium-dependent cellular processes such as pathogenesis.


Comparison of the transcriptional responses induced by acute morphine, methadone and buprenorphine.
Despite their widespread use in opioid maintenance treatment and pain management, little is known about the intracellular effectors of methadone and buprenorphine and the transcriptional responses they induce. We therefore studied the acute effects of these two opioids in rats, comparing our observations with those for the reference molecule, morphine. We determined the analgesic ED50 of the three molecules in the tail flick test, to ensure that transcriptional effects were compared between doses of equivalent analgesic effect. We analysed changes in gene expression over time in three cerebral structures involved in several opioid behaviours-the dorsal striatum, thalamus and nucleus accumbens-by real-time quantitative PCR. We analysed the expression of genes encoding proteins of the endogenous opioid system in parallel with that of Fos, a marker of neuronal activation. The acute transcriptional effects of methadone resembled those of morphine more closely than did those of buprenorphine, in terms of kinetics and intensities. Our results provide the first evidence that these two drugs widely used in pain management and opioid maintenance treatment can disturb the regulation of endogenous opioid system genes and induce molecular outcomes different from those observed with morphine.


UV254 induced sulfamethazine and carbamazepine elimination in the presence of nitrate: Roles of reactive species and generation risk of highly toxic DBPs.
This study systematically compared the degradation kinetics, conversion pathways, formation of disinfection by-products (DBPs), and changes in toxicity for sulfamethazine and carbamazepine in UV/nitrate system. Additionally, the study simulated the generation of DBPs in the post-chlorination process after the introduction of bromine ions (Br-). The contributions of UV irradiation, hydroxyl radicals (•OH), and reactive nitrogen species (RNS) to SMT degradation were determined to be 28.70 %, 11.70 %, and 59.60 %, respectively. The contributions of UV irradiation, •OH, and RNS to CBZ degradation were found to be 0.00 %, 96.90 %, and 3.10 %, respectively. A higher dosage of NO3- facilitated the degradation of both SMT and CBZ. Solution pH posed almost no effect on SMT degradation, while acidic conditions favored CBZ removal. The degradation of SMT was found to be slightly promoted at low concentrations of Cl-, while the presence of HCO3- significantly accelerated the degradation. Cl-, as well as HCO3-, retarded the CBZ degradation. Natural organic matter (NOM) as a free radical scavenger and UV irradiation filter posed a substantial inhibitory effect on the degradation of SMT and CBZ. The degradation intermediates and transformation pathways of SMT and CBZ by UV/NO3- system were further elucidated. The results showed that the main reaction pathways were bond-breaking reaction, hydroxylation, and nitration/nitrosation reaction. The acute toxicity of most of the intermediates generated during SMT and CBZ degradation was reduced after UV/NO3- treatment. After treatment of SMT and CBZ in UV/nitrate system, the DBPs generated in subsequent chlorination were mainly trichloromethane and a small amount of nitrogen-containing DBPs. After bromine ions were introduced in UV/NO3- system, a large amount of the originally generated trichloromethane was converted to tribromomethane.


Biochemical markers in prostatic cancer.
The status of the biochemical markers explored for use in patients with carcinoma of the prostate is reviewed. No unique marker has been identified. However, a variety of substances, primarily enzymes and specific proteins, found in the serum, urine, and prostatic fluid, have been evaluated. Their main value remains in the staging of disease and the monitoring of response to therapy.


Influence of food on the absorption of theophylline administered in the form of sustained release tablet and microgranules.
Two sustained-release formulations of theophylline, tablets (T) and microgranules (MG) forms, were administered in a randomized order to 8 healthy subjects in fasting or with a high-protein test meal (50 per cent). Blood was collected for 32h post-dose. In fasting subjects, absorption of theophylline was significantly faster for T (tmax 5 h) as compared with MG (tmax 8 h, p less than 0.05), but Cmax and AUC were comparable; intersubject variability was higher with T. Administration of a high-protein test meal with T produced a significant decrease of the zero-order absorption rate constant of theophylline (K omicron 37.8 +/- 9.1 mgh-1 after meal versus 58.8 +/- 13 mgh-1 in fasting, p = 0.01), tmax was doubled to 10 h, and Cmax increased by 25 per cent (6.33 +/- 2.16 mgl-1 versus 5.04 +/- 1.28 mgl-1, p less than 0.02); with MG, tmax were the same (8 h), Cmax were not significantly increased (4.79 +/- 0.84 mgl-1 versus 4.55 +/- 0.67 mgl-1), absorption was delayed (lag-time 1.28 +/- 0.58 h) and the absorption was slightly accelerated (K omicron 50.4 +/- 10.4 mgh-1 versus 42.3 +/- 11.9 mgh-1, NS). For each form bioavailability was not significantly modified by food. This study demonstrated that food rich in protein modifies the absorption rate of theophylline in a sustained-release tablet formulation but is without influence in a pH-independent, sustained-release microgranule formulation.


Effects of an Educational Book on Paediatric Oral Health knowledge in a sample of Italian women.
Parents and caregivers, particularly in Italy, often have limited knowledge about their children's oral health. The primary objective of the study is to evaluate the educational effectiveness of a book on nutrition and prevention of oral diseases entitled "Oral health of mother and child in the first 1000 days of life". The sample for this study was composed of 103 adult Italian women who were potential caregivers of one or more children (e.g., mothers, grandmothers, babysitters, and educators). The enrolled women completed a preliminary online survey which included questions about their socio-demographic characteristics and their knowledge on oral health in the first 1000 days of life of newborns (30 questions). Following the survey, they received the educational book. After reading it, the participants completed a second online survey with the same 30 questions, to measure any improvement in their knowledge. It appears that our educational book about nutrition and prevention of oral diseases was effective in enhancing knowledge among the participants in our study. These findings suggest that this educational resource has the potential to be a valuable tool in preventing oral diseases in paediatric populations. However, further confirmation of these results should be obtained through randomised controlled trials.


Drug interactions in the reinforcing effects of over-the-counter cough syrups.
Drug interactions in reinforcin effects of over-the-counter cough syrups were investigated by utilizing place preference conditioning in rats. Dihydrocodeine (2 mg/kg, IP) induced a small, non-significant place preference. On the other hand, concurrent dosing of dihydrocodeine (2 mg/kg, IP) and a mixture (SC) of methylephedrine (4 mg/kg), caffeine (4 mg/kg) and chlorpheniramine (0.8 mg/kg) produced a significant place preference, the mean conditioning score in this group being about 3 times higher than that in the dihydrocodeine alone group. The potentiation of dihydrocodeine-conditioned place preference was observed by combination with chlorpheniramine (0.8 mg/kg, SC) alone as well as with the mixture, but neither with methylephedrine (4 mg/kg, SC) nor with caffeine (4 mg/kg, SC). Chronic infusion of the dopamine D1 receptor antagonist SCH23390 (1.0 mg/kg/day, SC) during conditioning abolished the appetitive effects of dihydrocodeine combined with chlorpheniramine. In conclusion, it is suggested that the potentiation of appetitive effects of dihydrocodeine is mostly due to chlorpheniramine among three ingredients in the cough syrups, and that the dopaminergic system, especially D1 receptor, may play an important role in the potentiation effect of chlorpheniramine on the reinforcing effects of dihydrocodeine.


Upregulation of microRNA-143-3p induces apoptosis and suppresses proliferation, invasion, and migration of papillary thyroid carcinoma cells by targeting MSI2.
As a tumor-associated biological molecule, microRNA-143-3p (miR-143-3p) is implicated in the progression of papillary thyroid carcinoma (PTC). We conducted this study to elucidate the effects of miR-143-3p mediated by Musashi RNA binding protein 2 (MSI2) on the biological activities of PTC cells. The K1 cells with the lowest miR-143-3p expression were selected for the experiments. The targeting relationship between miR-143-3p and MSI2 was verified. The biological functions of miR-143-3p and MSI2 with respect to K1 cell proliferation, cycle distribution, apoptosis, invasion, migration, and tumorigenesis were studied using gain- and loss-of-function assays both in vitro and in vivo. MSI2 was verified to be a target gene of miR-143-3p. Cells treated with upregulation of miR-143-3p or silencing of MSI2 exhibited significantly decreased the expression of Bcl-2, PCNA, MCM2, Ki67, MSI2, MMP-2, and MMP-9. This was accompanied by inhibited cell proliferation, cell invasion, and migration, as well as a significant increase in Bax expression, cell cycle arrest, and cell apoptosis. More importantly, the tumor inhibitory effects of upregulated miR-143-3p were also confirmed in the tumor xenografts in nude mice. Our results indicate that upregulation of miR-143-3p suppresses the progression of PTC by impeding cell growth, invasion, and migration via downregulation of MSI2, highlighting the potential of miR-143-3p as a target for future PTC treatment.


Thiotungstate-copper interactions II. The effects of tetrathiotungstate on systemic copper metabolism in normal and copper-treated rats.
The intraperitoneal administration of tetrathiotungstate to rats (6-17.4 mg W/Kg BW) caused profound changes in copper metabolism in both normal rats and in rats pretreated with copper. Plasma copper associated with albumin increased, liver copper, particularly cytosol copper, was depleted, and biliary excretion was increased. There was also a movement of copper to higher molecular weight proteins in both liver cytosol and bile. In contrast to penicillamine, tetrathiotungstate did not increase liver cytosolic apometallothionein levels and reduced the rise provoked by copper. Metallothionein-bound copper was removed. Ceruloplasmin oxidase activity was inhibited and there was evidence for increased movement of copper into subcellular organelles, probably lysosomes. It is concluded that tetrathiotungstate has a genuine "decoppering" effect and could be considered as an alternative to thiomolybdates in the treatment of copper storage diseases.


Effects of equinatoxin II on isolated guinea pig taenia caeci muscle contractility and intracellular Ca2+.
Equinatoxin II (EqT II) is a approximately 20kDa cytotoxic and cytolytic protein isolated from the sea anemone Actinia equina. When injected intravenously to rats the toxin has been reported to produce a rapid cardiorespiratory arrest. In the present study, we show that EqT II increases the tension of spontaneous contractions and induces long-lasting contracture of guinea pig taenia caeci muscle. In taenia caeci, dissociated smooth muscle cells, microspectrofluorometric measurements, using the Ca(2+) indicator fura-2/AM, revealed that the toxin causes a marked increase in intracellular calcium, provided Ca(2+) is present in the external medium. The increase in intracellular Ca(2+) by EqT II was not blocked or diminished by the calcium channel blocker verapamil. Furthermore, pre-treatment of smooth muscle cells with Ca(2+)-ATPase inhibitor thapsigargin, or exposure of the cells to a high K(+) (75 mM) medium did not prevent EqT II-induced intracellular Ca(2+) increases. Replacement of external sodium by sucrose markedly modified the time course of Ca(2+) signals suggesting the involvement of the Na(+)/Ca(2+) exchanger in EqT II action. Our results strongly suggest that EqT II-induced increase in intracellular Ca(2+) and muscle tension are both dependent on the ability of EqT II to insert into the membrane and form pores allowing Ca(2+) influx into the cells. To our knowledge this is the first report showing that EqT II causes contraction and contracture of taenia caeci muscles and increases intracellular Ca(2+) in smooth muscle cells.


Laponite Lights Calcium Flickers by Reprogramming Lysosomes to Steer DC Migration for An Effective Antiviral CD8+ T-Cell Response.
Immunotherapy using dendritic cell (DC)-based vaccination is an established approach for treating cancer and infectious diseases; however, its efficacy is limited. Therefore, targeting the restricted migratory capacity of the DCs may enhance their therapeutic efficacy. In this study, the effect of laponite (Lap) on DCs, which can be internalized into lysosomes and induce cytoskeletal reorganization via the lysosomal reprogramming-calcium flicker axis, is evaluated, and it is found that Lap dramatically improves the in vivo homing ability of these DCs to lymphoid tissues. In addition, Lap improves antigen cross-presentation by DCs and increases DC-T-cell synapse formation, resulting in enhanced antigen-specific CD8+ T-cell activation. Furthermore, a Lap-modified cocktail (Lap@cytokine cocktail [C-C]) is constructed based on the gold standard, C-C, as an adjuvant for DC vaccines. Lap@C-C-adjuvanted DCs initiated a robust cytotoxic T-cell immune response against hepatitis B infection, resulting in > 99.6% clearance of viral DNA and successful hepatitis B surface antigen seroconversion. These findings highlight the potential value of Lap as a DC vaccine adjuvant that can regulate DC homing, and provide a basis for the development of effective DC vaccines.


Racial disparities in sleep disturbances among patients with and without coronary artery disease: The role of clinical and socioeconomic factors.
To investigate differences in sleep quality by race in participants with and without a prior myocardial infarction (MI). Case-control study. Emory-affiliated hospitals in Atlanta, Georgia. Two hundred seventy-three individuals (190 Black) ≤60 years of age with a verified MI in the previous 8 months, and 100 community controls (44 Black) without a history of MI. Sleep quality was assessed using the Pittsburgh Sleep Quality Index (PSQI). Psychological factors were assessed using standardized questionnaires and clinical risk factors through medical history and chart review. A significant interaction existed between race and MI status on sleep quality (P= .01), such that Black individuals with a history of MI, but not controls, reported worse sleep quality than their non-Black counterparts. Among MI cases, being Black was independently associated with higher PSQI scores after adjusting for baseline demographics (B = 2.17, 95% confidence interval 1.17, 3.17, P = .006). Clinical risk factors, psychological factors and socioeconomic status (household income and years of education) all contributed equally to explain race-related disparities in sleep among MI cases. After further adjustment for these factors, the association was attenuated and no longer significant (B = 0.70, 95% confidence interval = -0.10, 1.21, P = .26). Black post-MI patients, but not healthy controls, have significantly poorer sleep quality than non-Blacks. This difference is driven by a combination of factors, including clinical risk factors, psychological factors as well as adverse socioeconomic conditions among Black individuals with MI.


Surgical treatment of benign lesions and pathologic fractures of the proximal femur in children.
Benign lesions of the proximal femur region, such as simple bone cysts, aneurysmal bone cysts, and fibrous dysplasia, are common in children. Benign lesions may cause pathologic fractures, limb length inequities, and growth disturbances. Differential diagnoses, e.g., malignant bone tumors and osteomyelitis, are sometimes difficult to rule out. We aimed to evaluate outcomes in children with benign lesions of the proximal femur treated with curettage, bone grafting, and plate fixation. In this retrospective study, we included 30 children (median age 10.5 years; range 1.1-17.8 years) suffering from bone cysts and tumor-like lesions of the proximal femur region treated between 2002 and 2018. We analyzed plain X-ray images and CT scans in all children and obtained MRI scans in a selected group of children (63.3%). We examined histopathologic biopsy results for all bone lesions before initiating treatment. Surgical management comprised tumor curettage with adjuvant high-speed drilling and allogenic bone grafting supplemented by bone graft substitutes before plate fixation. Median follow-up interval was 87 months (range 24-156 months). We evaluated the healing of lesions according to Capanna's classification and rated functional outcomes according to Merle d'Aubigné and Postel score. Overall, 25 of 30 (83.3%) patients were admitted to hospital because of a pathologic fracture. We diagnosed simple bone cysts in 15 (50.0%) patients, aneurysmal bone cysts in 7 (23.5%) patients, and fibrous dysplasia in 8 (26.5%) patients. Bone consolidation was achieved in 22 of 30 (73.3%) patients after a mean of 5 months (range 3-7 months). The main complication was recurrence of the lesion in 4 of 30 (13.3%) patients. With respect to the Merle d'Aubigné and Postel scores, 17 of 30 (56.7%) patients obtained an excellent result (18 points), while 12 (40.0%) patients had a good result (15-17 points) and only 1 (3.3%) patient had a fair result (14 points). Surgical treatment of bone cysts and tumor-like lesions of the proximal femur by local resection or destruction of the lesion, followed by filling the defect with bone graft material and internal stabilization represents a safe and effective treatment option in children. Therapeutic, retrospective comparative study-Level III.


Preparation of fenofibrate solid dispersion using electrospray deposition and improvement in oral absorption by instantaneous post-heating of the formulation.
A coaxial electrospray technique was applied to a poorly soluble drug, fenofibrate (FEN), to increase its bioavailability. A particulate core-shell solid dispersion was designed using poly(methacrylic acid-co-methyl methacrylate) (Eudragit L-100) as a shell material and poly(vinyl pyrrolidone) K12-17 as a dispersant for FEN in the core phase. Although 58% of FEN remained in the crystalline state in the electrosprayed formulation, the dissolution behavior was significantly improved due to decrease in particle size, decrease in crystallinity, and increase in dispersion efficiency. The formulation was subjected to post-heating at 100 °C for 30 s to transform the remaining crystals into the amorphous state to further improve the dissolution behavior. Oral bioavailability was also on the order of: heated formulation>intact formulation>crystalline FEN. Instantaneous heating significantly improved the performance of the formulation despite its simple procedure, and thus can be a powerful step to be incorporated in the formulation manufacturing process.


Attitudes of Filipino parents of children with Down syndrome on noninvasive prenatal testing.
Globally, there has been an increasing uptake of noninvasive prenatal testing (NIPT). In the Philippines, the test is currently available through private laboratories and can be availed by families who can afford the out-of-pocket cost. In a country where elective termination of pregnancy is not an option, the question arises as to the relevance of this testing, even among health professionals. This is an exploratory qualitative study that explored the attitudes of Filipino parents of children with Down syndrome (DS) toward NIPT using thematic analysis of in-depth interviews. Study participants acknowledged the value of NIPT in providing early diagnosis and, subsequently, emotional, mental, spiritual, and financial preparation. This said, they also emphasized that such early detection may cause anxiety and even thoughts of termination for some, despite abortion being against the law and predominant religious beliefs. For those undergoing NIPT and receiving positive results, study participants highlighted the need to receive proper and nonbiased counseling from both health professionals and parents who have children with DS.


Estimation of local scale dispersion from local breakthrough curves during a tracer test in a heterogeneous aquifer: the Lagrangian approach.
The local scale dispersion tensor, Dd, is a controlling parameter for the dilution of concentrations in a solute plume that is displaced by groundwater flow in a heterogeneous aquifer. In this paper, we estimate the local scale dispersion from time series or breakthrough curves, BTCs, of Br concentrations that were measured at several points in a fluvial aquifer during a natural gradient tracer test at Krauthausen. Locally measured BTCs were characterized by equivalent convection dispersion parameters: equivalent velocity, v(eq)(x) and expected equivalent dispersivity, [lambda(eq)(x)]. A Lagrangian framework was used to approximately predict these equivalent parameters in terms of the spatial covariance of log(e) transformed conductivity and the local scale dispersion coefficient. The approximate Lagrangian theory illustrates that [lambda(eq)(x)] increases with increasing travel distance and is much larger than the local scale dispersivity, lambda(d). A sensitivity analysis indicates that [lambda(eq)(x)] is predominantly determined by the transverse component of the local scale dispersion and by the correlation scale of the hydraulic conductivity in the transverse to flow direction whereas it is relatively insensitive to the longitudinal component of the local scale dispersion. By comparing predicted [lambda(eq)(x)] for a range of Dd values with [lambda(eq)(x)] obtained from locally measured BTCs, the transverse component of Dd, DdT, was estimated. The estimated transverse local scale dispersivity, lambda(dT) = DdT/U1 (U1 = mean advection velocity) is in the order of 10(1)-10(2) mm, which is relatively large but realistic for the fluvial gravel sediments at Krauthausen.


Cystic fibrosis with non-G551D gating mutations in Italy: Epidemiology and clinical characteristics.
Cystic fibrosis transmembrane conductance regulator (CFTR) gating mutations (GMs) result in CFTR that is present at the cell surface but nonfunctional. Patients with the G551D mutation, the most prevalent worldwide, have been well studied. Italian GM patients have mainly non-G551D mutations. We studied their epidemiology and clinical characteristics in the period spanning the pre/post ivacaftor introduction to the Italian market. Data from the Italian CF Registry were used to describe patients with GMs and compare them with F508del homozygous (F/F) patients. In total, 186 patients with GMs (median [range] age, 21.96 [0.13-63.38] years) were identified among the 5552 patients included in the study (3.3%). They had lower sweat chloride values at diagnosis than the F/F and a lower ratio of males. In the GM group, examining the data of the years 2012 and 2017 and comparing with F/F, lung infection by Staphylococcus aureus and diabetes became less prevalent, and better FEV1 and nutritional status were observed in 2017. The cross-sectional evaluation year-by-year from 2012 to 2017 of the GM group showed improving trends in lung function and body mass index, and the decreasing prevalence of diabetes compared with F/F. Longitudinal evaluation of GM patients showed improvement in percent predicted (pp)FEV1 and nutrition in the 2012-2017 period. These variations correspond to the introduction of treatment with the CFTR potentiator ivacaftor (2014/2015). Italian patients with GMs are few and are characterized by milder phenotypes than F/F patients. Improved outcomes are likely influenced by treatment with ivacaftor.


Vaporized perfluorocarbon inhalation attenuates primary blast lung injury in canines by inhibiting mitogen-activated protein kinase/nuclear factor-κB activation and inducing nuclear factor, erythroid 2 like 2 pathway.
Blast lung injury is associated with high morbidity and mortality. Vaporized perfluorocarbon (PFC) inhalation has been reported to attenuate acute respiratory distress syndrome in humans and animal models. However, the effect of vaporized PFC on blast lung injury is still unknown. In this study, we investigated the protective effects and potential underlying mechanisms of action of vaporized PFC on blast lung injury in a canine model. This was a prospective, controlled, animal study in adult male hybrid dogs randomized to sham, blast (B), blast plus mechanical ventilation (B + M), and blast plus PFC (B + P) groups. All groups except for the sham were exposed to blast wave. The B + P group was treated with vaporized PFC for 1.5 h followed by 5.5 h mechanical ventilation. B + M group received 7.5 h mechanical ventilation and B group was observed for 7.5 h. Blast lung injury was induced using a shock tube. Blood gas, inflammatory cytokines, and oxidative stress were measured. Expression of nuclear factor (NF)-κB activation, mitogen-activated protein kinase (MAPK) and nuclear factor, erythroid 2 like 2 (Nrf2) were measured using western blot. Lung injury observed after blast exposure was marked by increased histopathological scores, ratio of lung wet to dry weight. PFC treatment attenuated blast lung injury as indicated by histopathological scores and ratio of lung wet to dry weight. PFC treatment downregulated interleukin (IL)-6, tumor necrosis factor (TNF)-α, and malondialdehyde (MDA), and upregulated superoxide dismutase (SOD) activity. PFC also suppressed expression of MAPK/NF-κB and Nrf2 protein levels. Our results suggest that PFC attenuated blast-induced acute lung injury by inhibiting MAPK/NF-κB activation and inducing Nrf2 expression in dogs.


Effect of certain antiadrenergic agents on systolic time intervals in essential hypertension.
Systolic time intervals, including preejection period (PEP) and left ventricular ejection time (LVET), were studied in patients with permanent essential hypertension before and after intra-venous administration of propranolol 0.2 mg/kg (11 patients), clonidine 0.002 mg/kg (10 patients) and methyldopa 2 mg/kg (12 patients). With propranolol, diastolic blood pressure was unchanged and the heart rate decreased, whilst PEP and LVET were significantly prolonged (P less than 0.001). Clonidine caused a fall in blood pressure (P less than 0.001), heart rate was slightly reduced, PEP was prolonged (P less than 0.001) and there was a significant decrease in LVET at 5 min. With methyldopa, no significant effect was observed after intravenous injection, but 7 days oral administration produced the identical effect as clonidine. These observations suggest that certain antihypertensive drugs may impair left ventricular performance and depress myocardial contractility.


[Ultrastructural analysis of changes in the lung parenchyma in the initial stages of experimental sepsis].
Histological and ultrastructural alterations in the lung parenchyma of rats with sepsis induced by implantation of 3 X 10(10) microorganisms (P. aeruginosa) beneath the skin were studied. Microorganisms with micropili were detected in septal spaces and in the alveoli 10 hours after implantation. Their effects were linked with microcirculatory alterations and destruction of endotheliocytes, alveolocytes and septal cells.


Single-step electrical field strength screening to determine electroporation induced transmembrane transport parameters.
The design of effective electroporation protocols for molecular delivery applications requires the determination of transport parameters including diffusion coefficient, membrane resealing, and critical electric field strength for electroporation. The use of existing technologies to determine these parameters is time-consuming and labor-intensive, and often results in large inconsistencies in parameter estimation due to variations in the protocols and setups. In this work, we suggest using a set of concentric electrodes to screen a full range of electric field strengths in a single test to determine the electroporation-induced transmembrane transport parameters. Using Calcein as a fluorescent probe, we developed analytical methodology to determine the transport parameters based on the electroporation-induced pattern of fluorescence loss from cells. A monolayer of normal human dermal fibroblast (NHDF) cells were pre-loaded with Calcein and electroporated with an applied voltage of 750V with 10 and 50 square pulses with 50μs duration. Using our analytical model, the critical electric field strength for electroporation was found for the 10 and 50 pulses experiments. An inverse correlation between the field strength and the molecular transport time decay constant, and a direct correlation between field strength and the membrane permeability were observed. The results of this work can simplify the development of electroporation-assisted technologies for research and therapies.


Vascular smooth muscle cell migration induced by domains of thrombospondin-1 is differentially regulated.
Thrombospondin-1 (TSP-1) stimulates vascular smooth muscle cell (VSMC) migration via defined intracellular signaling pathways. The aim of this study was to examine the signaling pathways whereby TSP-1 folded domains (amino-terminal [NH(2)], procollagen homology [PCH], all 3 type 1 repeats [3TSR], and a single recombinant protein containing the 3rd type 2 repeat, the type 3 repeats, and the carboxyl-terminal [E3T3C1]) induce VSMC migration. Quiescent VSMCs were pretreated with serum-free media or inhibitors: PP2 (c-Src), LY294002 (phosphatidylinositol 3-kinase), FPT (Ras), Y27632 (Rho kinase), SB202190 (p38 kinase), and PD98059 (extracellular signal-regulated kinase). Migration induced by serum-free media, TSP-1, NH(2), PCH, 3TSR, and E3T3C1 was assessed using a modified Boyden chamber. TSP-1, NH(2), 3TSR, and E3T3C1 induced VSMC chemotaxis (P < .05), but PCH did not (P > .05). PP2, FPT, SB202190, and PD98059 attenuated chemotaxis stimulated by TSP-1, NH(2), 3TSR, and E3T3C1 (P < .05). LY294002 inhibited TSP-1-induced and E3T3C1-induced (P < .05) but not NH(2)-induced or 3TSR-induced (P > .05) chemotaxis. Y27632 inhibited NH(2)-induced, 3TSR-induced, and E3T3C1-induced (P < .05) but not TSP-1-induced (P > .05) induced chemotaxis. TSP-1 folded domains are differentially dependent on intracellular signaling pathways to induce migration.


[Preliminary study on the antimutagenesis of vegetables and fruits].
Possible antimutation of 108 preparations of 91 kinds of vegetables and fruits on Salmonella typhimurium TA98 and TA100 mutants was tested. 4-nitroquinoline-N-oxide (4 NQO), N-methyl-N-nitro-N-nitrosoguanidine (MNNG), aflatoxin B1 (AFB1) and benzo (a) pyrene (BaP) were used as mutagens. The results showed that 67 (62%) preparations had antimutagenic action in vitro to different degrees. 9.6% of all preparations showed inhibition action on 4 NQO in TA100 mutant and 12.3%, in TA98, 5% on MNNG in TA100, 38% on AFB1 in TA100 and 45.1% in TA98, 28.9% on BaP in TA100. This experiment provides a scientific basis to the study of food resources as prevention of carcinogenesis.


A review of the biomaterials technologies for infection-resistant surfaces.
Anti-infective biomaterials need to be tailored according to the specific clinical application. All their properties have to be tuned to achieve the best anti-infective performance together with safe biocompatibility and appropriate tissue interactions. Innovative technologies are developing new biomaterials and surfaces endowed with anti-infective properties, relying either on antifouling, or bactericidal, or antibiofilm activities. This review aims at thoroughly surveying the numerous classes of antibacterial biomaterials and the underlying strategies behind them. Bacteria repelling and antiadhesive surfaces, materials with intrinsic antibacterial properties, antibacterial coatings, nanostructured materials, and molecules interfering with bacterial biofilm are considered. Among the new strategies, the use of phages or of antisense peptide nucleic acids are discussed, as well as the possibility to modulate the local immune response by active cytokines. Overall, there is a wealth of technical solutions to contrast the establishment of an implant infection. Many of them exhibit a great potential in preclinical models. The lack of well-structured prospective multicenter clinical trials hinders the achievement of conclusive data on the efficacy and comparative performance of anti-infective biomaterials.


Differential effect of dehydration on the voluntary activity of a diurnal toad.
Anuran amphibians' ability to maintain their activity at high temperatures or low humidity depends on their capacity to face dehydration, especially when they display diurnal and terrestrial life habits. Melanophryniscus rubriventris is a diurnal and terrestrial toad from humid Yungas Andean forests that breeds in temporary ponds. It is exposed to the recurrent risk of dehydration because of pond desiccations during the breeding season. Here, we study how M. rubriventris males behaviorally respond to dehydration by measuring their voluntary activity under an ex-situ experiment. Toads with different hydration levels were exposed to a circular track for voluntary activity measurements. Dehydrated males of M. rubriventris toads did not adopt a water-conserving posture staying active during the test and increasing walking under severe dehydration. Certain tolerance to dehydration would allow performing daily activities under challenging diurnal conditions. The increased walking under severe dehydration suggests water or shelter-seeking behavior that would be crucial for diurnal and terrestrial toads to overcome the unpredictable hydric environment during the breeding season.


Mesenchymal stem cells after the proprocessing of tanshinone IIA attenuate cognitive deficits and oxidative stress injury in an amyloid β-peptide (25-35)-induced rodent model of Alzheimer's disease.
To verify whether mesenchymal stem cells cocultured with tanshinone IIA may ameliorate Alzheimer's disease by inhibiting oxidative stress. Sixty male Sprague-Dawley rats were randomly divided into 4 groups named Sham, Aβ25-35, mesenchymal stem cells, and mesenchymal stem cells (tanshinone IIA). The rats were treated according to different groups. The neurobehavioral performance of Sprague-Dawley rats was evaluated via Morris water maze test. Histological changes were checked via hematoxylin-eosin staining. The levels of total antioxidant activity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and malondialdehyde in hippocampus were assayed by ELISA kit. The levels of Aβ, p-tau/tau, and p-AMP-activated protein kinase/AMP-activated protein kinase in hippocampus were checked by Western blot. Our research showed that the injection of mesenchymal stem cells (tanshinone IIA) into the hippocampus alleviated learning and memory deficits and reduced hippocampal neuronal injury in the Alzheimer's disease rats. Moreover, mesenchymal stem cells (tanshinone IIA) treatment suppressed oxidative stress, attenuated Aβ accumulation reduced Tau hyperphosphorylation, and enhanced the activity of AMP-activated protein kinase in the hippocampus of the Alzheimer's disease rats. However, there were almost no significant difference between the mesenchymal stem cells and Aβ25-35 groups. Mesenchymal stem cells (tanshinone IIA) transplantation may be a potential treatment for curing Alzheimer's disease, which may be related to the inhibition of oxidative stress.


No positive association between adrenergic receptor variants of alpha2cDel322-325, beta1Ser49, beta1Arg389 and the risk for heart failure in the Japanese population.
We investigated the correlation of adrenergic receptor polymorphisms, alpha(2c)Del322-325, beta(1)Ser49Gly and beta(1)Arg389Gly, with the risk of heart failure in the Japanese population. These polymorphisms were analysed by polymerase chain reaction-restriction fragment length polymorphism in patients with chronic heart failure due to idiopathic dilated cardiomyopathy (DCM) and compared with the control group. There were no differences or any trends in the allele and genotype frequencies of the beta(1)Ser49Gly and beta(1)Arg389Gly polymorphisms. The allele frequency of the alpha(2c)Del322-325 variant was lower in patients than in controls (0.11 vs. 0.04, P = 0.011 < 0.017, by Bonferroni correction), while the genotype frequency just failed to reach significance (P = 0.022 > 0.017, by Bonferroni correction). In this population, the variants beta(1)Ser49, beta(1)Arg389, and alpha(2c)Del322-325 do not appear to be risk factors for chronic heart failure due to DCM. The alpha(2c)Del322-325 variant may in fact confer some protection.


Assessment of the encapsulation effect of phenolic compounds from Spirulina sp. LEB-18 on their antifusarium activities.
This study aimed to investigate the antifungal activity of liposomal systems containing Spirulina sp. LEB-18 phenolic extract (PE) against Fusarium graminearum (Fg) isolates. The interaction between PE and phosphatidylcholine-based liposomes was monitored by HATR-FTIR, NMR, DSC, and cryo-TEM. After encapsulation, the active principle was released slower than the free PE, a fact that makes the former very promising as a natural antifungal. The PE encapsulation in the liposomes was responsible for changes in the dynamics of specific regions. These compounds affected the membrane hydration degree, ordered the lipid phosphate region and increased the disorder of the acyl chain methylenes. These physico-chemical effects may be related to the strong inhibition of four Fg isolates. Results were discussed by correlating structural similarities, as well as the membrane effects of the PE under study on antifusarium activities, and those found in the literature, thus enabling the PE mechanisms of action to be analyzed.


Regulation of the anaerobic metabolism in Bacillus subtilis.
The Gram-positive soil bacterium Bacillus subtilis encounters changing environmental conditions in its habitat. The access to oxygen determines the mode of energy generation. A complex regulatory network is employed to switch from oxygen respiration to nitrate respiration and various fermentative processes. During adaptation, oxygen depletion is sensed by the [4Fe-4S](2+) cluster containing Fnr and the two-component regulatory system ResDE consisting of the membrane-bound histidine kinase ResE and the cytoplasmic ResD regulator. Nitric oxide is the signal recognized by NsrR. Acetate formation and decreasing pH are measured via AlsR. Finally, Rex is responding to changes in the cellular NAD(+)/NADH ration. The fine-tuned interplay of these regulators at approximately 400 target gene promoters ensures efficient adaptation of the B. subtilis physiology.


The Pacific Northwest Harmful Algal Blooms Bulletin.
A bulletin communicating risk of toxic Pseudo-nitzschia blooms to shellfish harvest along the open coast of the Pacific Northwest region of the United States (the northeast Pacific Ocean spanning Washington and Oregon) is discussed. This Pacific Northwest Harmful Algal Blooms (PNW HAB) Bulletin is designed for shellfish managers with a focus on the razor clam fishery, but may also be informative to managers of the Dungeness crab fishery since domoic acid accumulation in crabs tends to lag accumulation in razor clams by a couple of weeks. The Bulletin complements beach phytoplankton monitoring programs by alerting coastal shellfish managers about adverse environmental conditions that could be conducive to a toxic Pseudo-nitzschia bloom. Beach monitoring programs are effective at determining when toxins have arrived at shellfish beaches, but a risk forecast based on near real-time biophysical information can provide managers with additional forewarning about potential future toxin outbreaks. Here, the approaches taken in constructing the risk forecasts, along with the reasoning and research behind them are presented. Updates to a historical PNW HAB Bulletin are described, as are the current workflow and the individual components of the updated Bulletin. Some successes and failures realized throughout the process are also pointed out for the benefit of the broader community. A self-assessment suggests that when the necessary data sources are available, the PNW HAB Bulletin provides an accurate forecast of risk associated with toxic Pseudo-nitzschia blooms. The Bulletin has proven beneficial to coastal shellfish managers by better informing decisions on sample collection, and harvest limits, openings, extensions, and closures.


Separation of major phospholipid classes by high-performance liquid chromatography and subsequent analysis of phospholipid-bound fatty acids using gas chromatography.
A sensitive high-performance liquid chromatographic method for the separation of major phospholipid (PL) classes in biological materials is described. Using this method it was easy to separate P-cholin, P-ethanolamine, P-serine, P-inositol, cardiolipin, sphingomyelin, lyso-P-choline and lyso-P-ethanolamine from skeletal and cardiac muscle samples. The method is based on the simultaneous use of a pH gradient and a polarity gradient. This procedure can easily be modified to optimize the separation of PLs from very different tissues. Subsequent analysis of the PL-bound fatty acids (FAs) by gas chromatography resulted in a well separated FA pattern. Following this FA separation it was possible to recalculate the specific PL content in the original sample.


Importance of pumice prophylaxis for bonding with self-etch primer.
Self-etching primers (SEP) have simplified the orthodontic bonding process, but questions have arisen regarding their reliability and efficiency. The goal of this study was to assess the importance of a pumice prophylaxis before bonding in reducing bond failures. Thirty orthodontic patients volunteered to participate in this prospective clinical trial. A split-mouth design was used; in each patient, 1 quadrant was randomly assigned to the pumice prophylaxis experimental group and the contralateral quadrant to the nonpumice group. A total of 508 teeth were bonded with SEP (Transbond Plus; 3M Unitek, Monrovia, Calif) and monitored for 3 months for bond failures. Thirty-five failures (6.9%) were recorded, with 6 (2.4%) in the pumice group and 29 (11.4%) in the nonpumice group. Chi-square analysis was used to compare bond failures as a total number between groups and also as the number of patients who experienced bond failures with each method. Statistically significant differences were found both in the total number of bond failures (P <.001) and in the number of patients with bond failures between groups (P <.01). A significantly lower and clinically acceptable bond failure rate was demonstrated with Transbond Plus SEP after pumice prophylaxis. This study produced strong evidence suggesting the need for pumice prophylaxis when using SEP for orthodontic bonding.


Epinephrine stimulates esophageal squamous-cell carcinoma cell proliferation via beta-adrenoceptor-dependent transactivation of extracellular signal-regulated kinase/cyclooxygenase-2 pathway.
Esophageal cancer is the sixth leading causes of cancer-related death in the world. It is suggested that beta-adrenoceptor is involved in the control of cell proliferation, but its role in the pathogenesis of esophageal cancer remains unknown. We therefore studied the role of beta-adrenergic signaling in the regulation of growth of an esophageal squamous-cell carcinoma cell line HKESC-1. Results showed that both beta(1)- and beta(2)-adrenoceptors were expressed in HKESC-1 cells. Stimulation of beta-adrenoceptors with epinephrine significantly increased HKESC-1 cell proliferation accompanied by elevation of intracellular cyclic AMP levels, which were abolished by beta(1)- or beta(2)-selective antagonists. Epinephrine also increased extracellular signal-regulated kinase-1/2 (ERK1/2) phosphorylation as well as cyclooxygenase-2 (COX-2) and cytosolic phospholipase A(2) expression, which were blocked by beta(1)- or beta(2)-selective antagonists. Moreover, epinephrine increased cyclin D(1), cyclin E(2), cyclin-dependent kinase (CDK)-4, CDK-6, and E(2)F-1 expression and retinoblastoma protein phosphorylation at Ser807/811, all of which were abrogated by beta(1)-adrenoceptor antagonist. Furthermore, epinephrine increased the expression of vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR)-1 and -2 in a beta(2)-adrenoceptor-, mitogen-activated protein kinase/ERK kinase (MEK)-, and COX-2-dependent manner. MEK or COX-2 inhibitor also significantly inhibited HKESC-1 cell proliferation induced by epinephrine. Collectively, we demonstrate that epinephrine stimulates esophageal squamous-cell carcinoma cell proliferation via beta-adrenoceptor-dependent transactivation of ERK/COX-2 pathway. Stimulation of beta(1)- and beta(2)-adrenoceptors also elicits a differential response on the expression of cell cycle regulators. These novel findings may shed new light on the understanding of beta-adrenergic signaling in the control of esophageal cancer cell growth.


Clinical features and long-term outcomes of patients with systemic polyarteritis nodosa diagnosed since 2005: Data from 196 patients.
The landscape of polyarteritis nodosa (PAN) has substantially changed during the last decades. Recent data regarding causes, characteristics, and prognosis of systemic PAN in the modern era are lacking. This retrospective study included patients with systemic PAN referred to the French Vasculitis Study Group between 2005 and 2019. Characteristics, associated conditions and outcomes were collected, and predictors of relapse and death were analyzed. 196 patients were included. Main clinical symptoms were constitutional (84%), neurological (59%), skin (58%) and musculoskeletal (58%) manifestations. Secondary PAN accounted for 55 (28%) patients, including myelodysplastic syndrome (9%), solid cancer (7%), lymphoma (4%) and autoinflammatory diseases (4%). No patient had active HBV infection. All treated patients (98.5%) received glucocorticoids (GCs), alone (41%) or in combination with immunosuppressants (59%), with remission achieved in 90%. Relapses were independently associated with age >65 years (HR 1.85; 95% CI1.12-3.08), gastrointestinal involvement (1.95; 95% CI1.09-3.52) and skin necrotic lesions (HR 1.95; 95%CI 1.24-3.05). One-, 5- and 10-year overall survival rates were 93%, 87% and 81%, respectively. In multivariate analyses, age >65 years (HR 2.80; 95%CI 1.23-6.37), necrotic purpura (HR 4.16; 95%CI 1.62-10.70), acute kidney injury (HR 4.89; 95% 1.71-13.99) and secondary PAN (HR 2.98; 95%CI 1.29-6.85) were independently associated with mortality. Landscape of PAN has changed during the last decades, with the disappearance of HBV-PAN and the emergence of secondary PAN. Relapse rate remains high, especially in aged patients with gastrointestinal and cutaneous necrosis, as well as mortality.


Antimicrobial Stewardship with Pharmacist Intervention Improves Timeliness of Antimicrobials Across Thirty-three Hospitals in South Africa.
Ensuring timely administration of antimicrobials is critical in the management of patients with infections. Mortality increases by 7.6% for every hour of delay in the administration of antimicrobial therapy in patients with sepsis. The time elapsed from the written antibiotic order to actual intravenous administration or 'hang-time' can often be several hours due to logistics within the hospital. Our purpose is to evaluate the change in compliance with administering antimicrobials within an hour of prescription after implementation of a national antibiotic stewardship pharmacist-driven hang-time process improvement protocol. This was a prospective multicenter study in 33 South African hospitals from 1 July 2013-30 August 2014. Two pilot sites established the mechanism for noninfectious disease pharmacists to make interventions and document hang-time data. Following this, a hang-time compliance assessment was initiated using the tools of healthcare improvement spread methodology. This consisted of five stages and an implementation toolkit was developed. The pharmacist study coordinator was responsible for implementation, the development of an implementation toolkit and real-time coordination of data with monthly feedback to all sites. A total of 32,985 patients who received intravenous antibiotics were assessed for hang-time compliance with first doses of new antibiotic orders. Over the 60-week period, 21,069 patients received antibiotics within an hour following prescription and were assessed as hang-time compliant. The change in improvement of hang-time compliance following implementation of a pharmacist-driven hang-time process improvement protocol was 41.2% pre-intervention week 1 (164/398) to 78.4% post-intervention week 60 (480/612; P < 0.0001). Pharmacists reviewed and evaluated twice as many patients during the final 4 weeks (1680) compared to the first 4 weeks (834; P < 0.0001). Noninfectious disease pharmacists can significantly improve the timely administration of antimicrobials and contribute to low-hanging-fruit antimicrobial stewardship initiatives within a hospital system in a resource-limited country.


Cartilage diversification and modularity drove the evolution of the ancestral vertebrate head skeleton.
The vertebrate head skeleton has evolved a myriad of forms since their divergence from invertebrate chordates. The connection between novel gene expression and cell types is therefore of importance in this process. The transformation of the jawed vertebrate (gnathostome) head skeleton from oral cirri to jointed jaw elements required a diversity of cartilages as well as changes in the patterning of these tissues. Although lampreys are a sister clade to gnathostomes, they display skeletal diversity with distinct gene expression and histologies, a useful model for addressing joint evolution. Specifically, the lamprey tissue known as mucocartilage has noted similarities with the jointed elements of the mandibular arch in jawed vertebrates. We thus asked whether the cells in lamprey mucocartilage and gnathostome joint tissue could be considered homologous. To do this, we characterized new genes that are involved in gnathostome joint formation and characterized the histochemical properties of lamprey skeletal types. We find that most of these genes are minimally found in mucocartilage and are likely later innovations, but we do identify new activity for gdf5/6/7b in both hyaline and mucocartilage, supporting its role as a chondrogenic regulator. Contrary to previous works, our histological assays do not find any perichondrial fibroblasts surrounding mucocartilage, suggesting that mucocartilage is non-skeletogenic tissue that is partially chondrified. Interestingly, we also identify new histochemical features of the lamprey otic capsule that diverge from normal hyaline. Paired with our new insights into lamprey mucocartilage, we propose a broader framework for skeletal evolution in which an ancestral soxD/E and gdf5/6/7 network directs mesenchyme along a spectrum of cartilage-like features.


Epidemiology and risk factors for febrile ureteral stent-associated urinary tract infections: A prospective observational cohort study.
We aimed to determine the incidence and risk factors of febrile ureteral stent-associated urinary tract infections (FUSAUTI). Hospitalized adult patients with ureteral stent (US) placement or exchange were prospectively enrolled. Patients with kidney transplantation of less than one year were excluded. Patients were followed until US removal/exchange or six months after inclusion. Out of 663 patients included in the study, 48 had at least one FUSAUTI (cumulative incidence 7.24%; 95% confidence interval [CI] 5.39-9.48). The incidence rate of FUSAUTI was 9.04 (95% CI 6.67-12.2) per 10,000 US-days. Ten patients (20.8%) experienced sepsis or septic shock. The most frequently isolated microorganisms were Escherichia coli (38%), Enterococcus spp. (14.5%), Candida spp. (9%) and Pseudomonas aeruginosa (9%). In multivariable logistic regression analysis, female gender, an age adjusted Charlson comorbidity index score> 3, an urethral stent placement concomitant with US placement, and a history of urinary tract infection within three months were significantly associated with a higher risk of FUSAUTI. After US placement, 7.24% of patients developed at least one FUSAUTI and, in a quarter of cases, a serious infection. Urethral stent placement was the only modifiable risk factor identified. Future interventional studies are needed to reduce FUSAUTI in these patients.


Lipopolysaccharide-induced suppression of periodontal ligament cell proliferation and apoptosis are strengthened under high glucose conditions.
The present study aimed to investigate the effect of lipopolysaccharide (LPS) on the proliferation and apoptosis of human periodontal ligament (hPDL) cells under normal glucose or high glucose conditions. Primary cultures of hPDL cells were prepared from extracted premolars of patients. The cells were incubated with 0, 1, or 10μg/mL LPS under normal glucose (5.5mmol/L) or high glucose (25mmol/L) conditions for 24h or 48h. Cell proliferation was detected using a CCK-8 assay, and cell apoptosis was measured by Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining. BCL2 and BAX mRNA and protein levels were measured by real-time polymerase chain reaction and western blotting, respectively. LPS (10μg/mL) induced significant inhibition of cell proliferation and cell apoptosis, and a significant decrease in the BCL2/BAX ratio in the cells cultured with 5.5mmol/L glucose. These effects of LPS were increased significantly in cells treated with 25mmol/L glucose. Analysis of variance of the factorial design revealed that high glucose and LPS had a significant interaction for cell apoptosis, but not for cell proliferation. High glucose augmented LPS-induced hPDL cell apoptosis and cell proliferation inhibition. LPS and high glucose might interact to induce cell apoptosis.


Enhancement of anti-neoplastic effects of cuminaldehyde against breast cancer via mesoporous silica nanoparticle based targeted drug delivery system.
Synthesis of novel drug delivery system for targeted delivery of cuminaldehyde to breast cancer cells and the subsequent analyses of anti-neoplastic potential of the drug. 3-carboxy-phenyl boronic acid (PBA) conjugated and polyacrylic acid (PAA) gated mesoporous silica nanoparticles (MSNs) were synthesized for the targeted delivery of cuminaldehyde (CUM) to breast cancer cells. Enhancement of anti-neoplastic effects of cuminaldehyde (4-isopropylbenzaldehyde) by the nanoconjugates was assessed. The anti-cancer effects of non-targeted and targeted drug-nanoconjugates were examined in vitro and in vivo. The targeted drug-nanoconjugates caused cell cycle arrest and induced the intrinsic pathway of apoptosis in MCF-7 cells through mitochondrial damage. In vivo intravenous injection of the targeted drug-nanoconjugates led to effective reduction in growth of 4 T1 induced mammary pad tumor in female BALB/c mice via augmented accumulation of cuminaldehyde. The drug-nanoconjugates did not exhibit any systemic toxicity. Therefore, MSN-PBA-CUM-PAA represents a potent therapeutic model for breast cancer treatment.


Development of the spinal nerves in the lamprey: II. Outflows from the spinal cord.
During a series of studies on the development of spinal nerves in the tail of larval lampreys (13 mm, 26 days), Lampetra japonica, we observed outflows of cytoplasmic processes (cytoplasmic outflow) or whole cell bodies (cellular outflow) from the neural tube or from the cord. Three types were distinguished according to their site of exit from the surface of the neuraxis. The dorsal outflow (DO) is the cellular outflow seen on the midsagittal surface of the dorsal wall of the caudalmost region of the neural tube, just rostral or caudal to the opening in the dorsal tube wall. It is hypothesized that the cells from the dorsal neural tube become polymorphous cells scattered in the extramedullar space. The dorsolateral outflow (DLO) is the cytoplasmic outflow emerging from the dorsolateral aspect of the spinal cord at the intermyotome level. DLO fibers are non-myelinated fibers arising from the dorsolateral tract (DLT) and, after piercing the basal lamina and the glia limitans, run in the myosepta laterally to spread along the deep surface of the dermis. DLO fibers terminate in two different ways: those of one group pierce the dermis and the basal lamina to end as intraepidermal free endings that contain aggregates of clear vesicles, and those of the other group form varicosities that lie within depressions on the lateral cell surface of myotomes. DLO fibers in the extramedullary space characteristically lack any sheaths, including the basal lamina. The ventrolateral outflow (VLO) represents primitive ventral roots consisting of both the cytoplasmic and the cellular outflows: the former shows the axonal outgrowth from primitive somatomotor neurons, and the latter is represented by elongated cells derived from the glia limitans, extending along the axonal processes to form the Schwann cell sheath around the proximal portion of the ventral root axons. The basal lamina covering the cord does not extend along the VLO fibers. The developing ventral root contains axons at different stages of differentiation. Some axons end on the medial surface of the myotome at the midsegmental level to form neuromuscular junctions. However, axonal processes of primitive motoneurons form close contacts with muscle cells by means of small cytoplasmic projections that contain no synaptic vesicles and lack the basal lamina interposed between them. The proposed sequence of development, based on stages in caudal to rostral sections, is the DO, the DLO, and then the VLO.(ABSTRACT TRUNCATED AT 400 WORDS)


Goldenhar syndrome: a case report with review of literature.
A case of Goldenhar Syndrome, in an adult male, with the typical triad of auricular appendages, epibulbar dermoid and vertebral anomalies is presented. The relevent literature is review. The differential diagnosis of this Syndrome from a few similar syndromes is stressed.


Basic principles of nuclear medicine techniques for detection and evaluation of trauma and sports medicine injuries.
Nuclear medicine skeletal imaging is a very sensitive technique for evaluating bone and muscle abnormalities because it can detect minor changes in metabolism and blood flow. The specificity of bone imaging, however, depends on the ability of the nuclear medicine physician to make a differential diagnosis. To aid in making a specific diagnosis, this article describes the various patterns of abnormality in stress fractures, tibial stress syndrome (shin splints), compartment syndrome, enthesopathy, and traumatic fractures. The characteristic scintigraphic appearance of joint injuries, muscle injuries (rhabdomyolysis), and radionuclide arthrography is discussed and the way the scan patterns change with time in these various disorders is described. A brief summary of the basic anatomy and physiology of bone and muscle in normal and injured tissue is presented and the basic mechanisms which cause the various abnormal scan patterns is postulated. In addition, a staging system for stress fractures is presented to help direct the referring physician toward the proper management of the injured patient. In most cases, nuclear medicine skeletal imaging can be used to differentiate between acute muscle injury, tibial stress syndrome, skeletal injury (periosteal reaction, stress fracture, and traumatic fracture) or an abnormality that is entirely associated with the joint or connective tissue. This differential diagnosis is easier if the nuclear medicine procedure is performed within a few days after the onset of injury.


delta-Aminolevulinic acid-synthesizing enzymes need an RNA moiety for activity.
When Chlamydomonas enzymes that convert glutamate to delta-aminolevulinic acid are separated into three fractions, one of the fractions contains RNA, and the RNA moiety is needed for the enzyme activity.


Azvudine and mortality in patients with coronavirus disease 2019: A retrospective cohort study.
Several studies have found that azvudine (FNC) can inhibit severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication both in vivo and in vitro. However, the effect of FNC on the risk of death in patients with coronavirus disease 2019 (COVID-19) is unclear. This study aims to investigate the effect of FNC on the risk of death in patients with coronavirus disease 2019 (COVID-19). Charts of consecutive patients hospitalized at five hospitals in Chongqing with confirmed COVID-19. The primary outcome of the study was 28-day mortality. Secondary outcomes were: ICU admission rates, length of hospital and ICU stay, and also the range of mechanical ventilation days when admission. We compared primary outcome in patients who received FNC with those in patients who did not, using a multivariable model with inverse probability weighting according to the propensity score. We included 1,110 patients in our study cohort. Of the 236 patients treated with FNC, 30 died within 28 days (12.7%), and of the 874 patients not treated with FNC, 206 died within 28 days (23.6%). In the crude, unadjusted analysis, a significant beneficial effect of FNC in terms of the 28-day mortality (OR 0.472, 95% CI 0.312-0.714; p < 0.001) in the overall population was detected. The adjusted odds ratio by multivariate analysis was (OR 0.498, 95% CI 0.287-0.864; p = 0.013). In the multivariate analysis with inverse probability weighting according to the propensity score, a significantly beneficial effect of FNC in terms of the 28-day mortality was further confirmed (OR 0.754, 95% CI 0.614-0.925; p = 0.007). Moreover, multivariable propensity-score analyses with matching also yielded similar results (OR 0.438, 95% CI 0.246-0.778; p = 0.005). Our results reveal that in patients with COVID-19, FNC administration was associated with a significantly reduced 28-day mortality.