Contribution of ENPP1, TCF7L2, and FTO polymorphisms to type 2 diabetes in mixed ancestry ethnic population of South Africa.
Transcription factor 7-like 2 gene (TCF7L2), fat mass and obesity-associated gene (FTO), and ectonucleotide pyrophosphatase/phosphodiesterase gene (ENPP1) are known risk loci for type 2 diabetes (T2DM) mostly in European populations. To assess the association of these genes with T2DM risk in a South African mixed-ancestry population. Five hundred and sixty six participants were genotyped for ENPP1-rs997509 and -rs1044498, FTO-9941349 and -rs3751812, TCF7L2-rs12255372 and -rs7903146 polymorphisms using Taqman genotyping assays and validated by automated sequencing to assess the association of the polymorphisms with cardiometabolic traits. In logistic regression models adjusted for age, sex, body mass index (BMI) and insulin resistance, minor allele of rs997509 was associated with a higher risk of prevalent T2DM under a recessive model [odd ratio 4.60 (95% confidence interval: 1.07 to 19.86); p = 0.040].Under additive model, the rs7903146 [1.43 (1.00 to 2.04); p= 0.053] and rs9941349 [1.43 (1.00 to 2.04); p = 0.052] minor alleles showed marginally significant associations with a high risk of T2DM. However, only the rs7903146 alleles (p=0.011) and genotypes (p=0.025) distributions were statistically significantly different between diabetic and non-diabetic individuals. Our findings demonstrate that ENPP1, TCF7L2, and FTO may predispose to T2DM in the mixed-ancestry population.


Is there scope for expanding the optometrist's scope of practice in Singapore? - A survey of optometrists, opticians in Singapore.
In Singapore, optometrists' roles are limited compared to their counterparts elsewhere. The purpose of the survey is to investigate optometrists' current roles, views on extended roles, self-reported primary eye care knowledge, needs for continuing professional education (CPE) and views on suitable modes for CPE. Members of the Optometrist and Optician Board (OOB) were invited via email to take part in an anonymous online survey. The survey questions covered the following areas: current scope of practice, self-rated primary eye care knowledge, confidence in screening, co-managing minor eye conditions, CPE and referral behavior. A total of 230 optometrists completed the survey (response rate 30%). Their current roles were limited to diagnostic refraction (92%), colour vision assessment (65%), contact lens fitting and dispensing (62%) amongst others. The average self-rated score for primary eye care knowledge was 8.2 ± 1.4; score range 1-10 (1-Very poor, 10-Excellent). Self-rated confidence scores for screening for cataract, diabetic retinopathy, chronic glaucoma and age-related macular degeneration were 2.7 ± 1.5, 3.7 ± 1.9, 4.0 ± 1.9 and 3.8 ± 1.8, respectively. 71% of the optometrists felt that they should undertake regular CPE to improve their primary eye care knowledge. Blended learning (eLearning and traditional face-to-face lectures) (46.1%) was the most preferred mode for CPE delivery. Optometrists in Singapore represent a skilled underutilized primary eye care provider. Though their self-reported primary eye care knowledge is high, their confidence in screening and co-managing chronic eye conditions is low. Enabling them for extended primary eye care role would require further training. Singapore ageing population has led to greater eye care demands. Task-shifting from ophthalmologists to optometrists has been proposed in the literature to handle this growing care demands. At this juncture, this study provides evidence based answers to issues revolving around optometrists' readiness for a role expansion in Singapore.


Myofibroblasts in focal nodular hyperplasia of the liver.
Myofibroblasts have been identified by electron microscopy in a case of Focal Nodular Hyperplasia (FNH) of the liver associated with oral contraceptives. The contractile fibroblasts were observed in the immediate vicinity of, or in close contact with, the proliferating bile ductules and in the recesses between parenchymal cells. Transitional forms between fat-storing cells (Ito cells), fibroblasts and myofribroblasts were observed. Transition of fat-storing cells to myofibroblasts, possibly under the influence of oestrogens, may be responsible for the fibrosis and retraction in FNH.


DNA ring closure mediated by protein HU.
The histone-like protein HU serves as an accessory factor that can facilitate the interaction of certain proteins with their specific DNA binding sites. Examples occur in different systems for prokaryotic DNA replication, transcription, and gene regulation. The protein-DNA interactions that are stimulated by HU generally involve coiling or looping of the DNA, and the possibility has been considered that HU exerts its effect by contributing flexibility to different DNA binding sites, but there has been no direct demonstration of this. To explore the possibility that HU can mediate tight DNA curvatures, we studied its effect on the formation of DNA circles when DNA ligase cyclizes short linear DNA fragments. It is demonstrated that HU greatly increases the cyclization rates of all fragments that were examined having lengths greater than 98 base pairs. Fragments of 99, 108, 120, or 126 base pairs could not cyclize in the absence of HU, but cyclization went rapidly with HU, showing that HU can mediate very tight DNA curvatures.


Specificity of autoantibodies to SS-A/Ro on a transfected and overexpressed human 60 kDa Ro autoantigen substrate.
The objective of this study was to analyze apparently discrepant results that arose during the use of an indirect immunofluorescence (IIF) assay using transfected HEp-2 cells to detect anti-SS-A/Ro autoantibodies in human sera. Fourteen sera that had SS-A/Ro antibodies as detected on this commercial substrate, but did not have antibodies to SS-A/Ro as determined by double immunodiffusion (ID) or enzyme-linked immunosorbent assay (ELISA), were studied by immunoprecipitation (IP) of radiolabeled cell extracts and full-length recombinant SS-A/Ro. A multi-antigen strip immunoblotting (IB) assay containing both the 52- and 60-kDa antigens was included in the analysis. We confirmed that 12 of 14 of the sera under study had antibodies to SS-A/Ro protein antigens as determined by at least one other immunoassay. One serum had antibodies to hyRNA but no detectable reactivity with the 52- or 60-kDa antigens. One serum remained negative in all assays for SS-A/Ro autoantibodies.


Splice variants of mitofusin 2 shape the endoplasmic reticulum and tether it to mitochondria.
In eukaryotic cells, different organelles interact at membrane contact sites stabilized by tethers. Mitochondrial mitofusin 2 (MFN2) acts as a membrane tether that interacts with an unknown partner on the endoplasmic reticulum (ER). In this work, we identified the MFN2 splice variant ERMIT2 as the ER tethering partner of MFN2. Splicing of MFN2 produced ERMIT2 and ERMIN2, two ER-specific variants. ERMIN2 regulated ER morphology, whereas ERMIT2 localized at the ER-mitochondria interface and interacted with mitochondrial mitofusins to tether ER and mitochondria. This tethering allowed efficient mitochondrial calcium ion uptake and phospholipid transfer. Expression of ERMIT2 ameliorated the ER stress, inflammation, and fibrosis typical of liver-specific Mfn2 knockout mice. Thus, ER-specific MFN2 variants display entirely extramitochondrial MFN2 functions involved in interorganellar tethering and liver metabolic activities.


[Role of tropomyosin in the sensitivity of (Na+ + K+) ATPase to ouabain].
EDTA treatment of isolated plasma membranes from MF2S cells increased 1,000 fold the sensitivity of (Na+ + K+) ATPase activity to ouabain. The original sensitivity of the enzyme to the drug is recovered after addition of tropomyosin together with Ca++ ions to the treated membranes.


Solutions that enable ablative radiotherapy for large liver tumors: Fractionated dose painting, simultaneous integrated protection, motion management, and computed tomography image guidance.
The emergence and success of stereotactic body radiation therapy (SBRT) for the treatment of lung cancer have led to its rapid adoption for liver cancers. SBRT can achieve excellent results for small liver tumors. However, the vast majority of physicians interpret SBRT as meaning doses of radiation (range, 4-20 Gray [Gy]) that may not be ablative but are delivered within about 1 week (ie, in 3-6 fractions). Adherence to this approach has limited the effectiveness of SBRT for large liver tumors (>7 cm) because of the need to reduce doses to meet organ constraints. The prognosis for patients who present with large liver tumors is poor, with a median survival ≤12 months, and most of these patients die from tumor-related liver failure. Herein, the authors present a comprehensive solution to achieve ablative SBRT doses for patients with large liver tumors by using a combination of classic, modern, and novel concepts of radiotherapy: fractionation, dose painting, motion management, image guidance, and simultaneous integrated protection. The authors discuss these concepts in the context of large, inoperable liver tumors and review how this approach can substantially prolong survival for patients, most of whom otherwise have a very poor prognosis and few effective treatment options. Cancer 2016;122:1974-86. © 2016 American Cancer Society.


The genomic landscape of cholangiocarcinoma reveals the disruption of post-transcriptional modifiers.
Molecular variation between geographical populations and subtypes indicate potential genomic heterogeneity and novel genomic features within CCA. Here, we analyze exome-sequencing data of 87 perihilar cholangiocarcinoma (pCCA) and 261 intrahepatic cholangiocarcinoma (iCCA) cases from 3 Asian centers (including 43 pCCAs and 24 iCCAs from our center). iCCA tumours demonstrate a higher tumor mutation burden and copy number alteration burden (CNAB) than pCCA tumours, and high CNAB indicates a poorer pCCA prognosis. We identify 12 significantly mutated genes and 5 focal CNA regions, and demonstrate common mutations in post-transcriptional modification-related potential driver genes METTL14 and RBM10 in pCCA tumours. Finally we demonstrate the tumour-suppressive role of METTL14, a major RNA N6-adenosine methyltransferase (m6A), and illustrate that its loss-of-function mutation R298H may act through m6A modification on potential driver gene MACF1. Our results may be valuable for better understanding of how post-transcriptional modification can affect CCA development, and highlight both similarities and differences between pCCA and iCCA.


Identification by Random Mutagenesis of Functional Domains in KREPB5 That Differentially Affect RNA Editing between Life Cycle Stages of Trypanosoma brucei.
KREPB5 is an essential component of ∼ 20S editosomes in Trypanosoma brucei which contains a degenerate, noncatalytic RNase III domain. To explore the function of this protein, we used a novel approach to make and screen numerous conditional null T. brucei bloodstream form cell lines that express randomly mutagenized KREPB5 alleles. We identified nine single amino acid substitutions that could not complement the conditional loss of wild-type KREPB5. Seven of these were within the RNase III domain, and two were in the C-terminal region that has no homology to known motifs. Exclusive expression of these mutated KREPB5 alleles in the absence of wild-type allele expression resulted in growth inhibition, the loss of ∼ 20S editosomes, and inhibition of RNA editing in BF cells. Eight of these mutations were lethal in bloodstream form parasites but not in procyclic-form parasites, showing that multiple domains function in a life cycle-dependent manner. Amino acid changes at a substantial number of positions, including up to 7 per allele, allowed complementation and thus did not block KREPB5 function. Hence, the degenerate RNase III domain and a newly identified domain are critical for KREPB5 function and have differential effects between the life cycle stages of T. brucei that differentially edit mRNAs.


A review of mathematical models for composting.
Composting is a valuable method to treat and valorize organic waste. However, the process is defined by its dynamic nature and governed by a multitude of operating parameters. As such, mathematical modelling of the process offers a powerful tool to simulate and predict the variable outcomes of the process, allowing for its optimization. This can include improving efficiency, lowering costs and reducing environmental impact. To aid with the development of future models, we provide an up to date review and assessment on the state of the art of composting modelling. By reviewing 40 years of literature, this review paints the most complete picture of the field to date. This includes an analysis of trends in composting modelling: looking at the type of systems that are targeted, the aim of the models and the approaches to kinetics and mass and heat transfer. Regarding modelling approaches, we explore the fractionation of both substrates and microorganisms, the biological processes that can be included (disintegration, hydrolysis, uptake and death) and their kinetics (first-order, Monod-type), energy balances (biological generation, convection, conduction) and mass balances. We also provide a review of the results of sensitivity analyses performed on composting models, finding that models are most sensitive to microbial growth and death rates, as well as consumption rates and product yields. In the final portion of the review, we identify, explore, and provide guiding recommendations for work on emerging areas and areas requiring development in composting modelling (volume change, pH, maturation, artificial intelligence, etc.).


Pulmonary function after laparoscopic and open cholecystectomy.
Laparotomy causes a significant reduction of pulmonary function, and atelectasis and pneumonia occur after open cholecystectomy. In this prospective, randomized study, we evaluated the hypothesis that pulmonary function is less restricted after laparoscopic cholecystectomy (LC) than after open cholecystectomy (OC). Sixty patients underwent laparoscopic (n = 30) or open (n = 30) cholecystectomy. The two groups did not differ significantly in age, sex, intraoperative findings, and preoperative pulmonary function. Pulmonary function tests, arterial blood-gas analysis, and chest radiographs were obtained in both groups before operation and on postoperative day 1. The forced expiratory volume in 1 s (mean +/- SD values; OC, 1.49 +/- 0.77 L/s; LC, 2.33 +/- 0.80 L/s; p > 0.0001) and the forced vital capacity (OC, 2.40 +/- 0.66 L; LC, 2.93 +/- 1.05 L; p > 0.01) were more suppressed in patients having OC than in those having LC. Similar results were found for the peak expiratory flow (OC, 3.51 +/- 1.35 L/s; LC, 4.27 +/- 1.66 L/s; p > 0.05), expiratory reserve volume (OC, 0.73 +/- 0.34 L; LC, 0.92 +/- 0.43 L; p > 0.05), and the midexpiratory phase of forced expiratory flow (FEF25-75) (OC, 1.45 +/- 0.54 L/s; LC, 1.60 +/- 0.73 L/s; NS). Laparoscopic cholecystectomy was associated with a significantly lower incidence of (30 vs 70%) and less severe atelectasis and better oxygenation. Pulmonary function is better preserved after LC than after OC.


Postinfarction ventricular aneurysmectomy.
Most operations performed for the treatment of ventricular aneurysm do not achieve maximal rehabilitation of the damaged heart. Cardiac surgeons generally ignore the importance of the flail septum that results from anteroseptal infarction. Many believe that the obstructed left anterior descending coronary artery must be carefully avoided during closure of the ventriculotomy incision. In addition, many surgeons believe that it is necessary to buttress all ventricular sutures with Teflon. For some reason, there seems to be a fear that the left ventricular volume will be reduced to an intolerable level after proper ventricular aneurysmectomy. Between January 1976 and December 1982, 102 patients underwent ventricular aneurysmectomy at St. Joseph's Hospital Health Center. The hospital mortality rate was 5.9 percent. The operative technique described emphasizes the need for foreshortening the fibrosed septum in an effort to minimize residual paradoxic motion. The left anterior descending coronary artery is routinely incorporated in the eversion technique; Teflon buttressing is never employed. Our surgical technique has evolved from a surgical experience that began in 1962 at the Cleveland Clinic Hospital.


The Quality of Sex Life Among Patients Before and After Lung Transplantation: A Single-Center Study.
Lung transplantation (LTx) is the only effective method of treatment for patients with end-stage lung diseases; LTx prolongs and increases the quality of life (QoL). An important aspect of QoL that changes in the course of severe diseases is the quality of sex life. This aspect is yet to be discussed in relationship to LTx. We aim to compare patients' quality of sex life at the qualification process with patients' who underwent LTx. The studied group consisted of 100 patients (24 women before and 16 after LTx, 39 men before and 21 after LTX) who were admitted to the lung transplantology department for qualification or to control the function after LTx. To assess the patients' quality of sex life, we used The Changes in Sexual Functioning Questionnaire (CSFQ) and World Health Organization (WHO) QoL-BREF. To assess lung function, patients underwent a 6-Minute-Walk-Test (6MWT). Patients after LTx obtained higher results-compared to patients qualified for LTx-in the WHO QoL-BREF in every domain (somatic, psychological, social, and environment). Men after LTx got more points in every domain and better total score (53 ± 5.62 vs 44.23 ± 10.28 point; P < .05) in CSFQ. Women before and after LTx obtained comparable results in CSFQ. Results of 6-Minute-Walk-Test were better among patients after LTx than in qualified patients (523.62 ± 95.71 vs 333.14 ± 145.38 and 524.12 ± 56.17 vs 317.20 ± 141.6, respectively for men and women). Patients after LTx show better pulmonary function and quality of sex life than qualified. Preliminary results encourage us to conduct research on a larger group.


Effect of gum Arabic on aminopyrine demethylation in rats.
Stimulation of aminopyrine demethylation induced in rats by oral or i.p. administration of phenobarbital was partially inhibited in animals receiving daily treatments of 2 x 200 mg/kg gum Arabic p.o.


Nongenomic regulation of chondrocyte membrane fluidity by 1,25-(OH)2D3 and 24,25-(OH)2D3 is dependent on cell maturation.
1,25-(OH)2D3 and 24,25-(OH)2D3 regulate rat costochondral chondrocyte cultures in a metabolite-specific manner; 1,25-(OH)2D3 targets primarily growth zone cells (GC) and 24,25-(OH)2D3 targets primarily resting zone cells (RC). Some of the effects are nongenomic, since incubation of isolated membrane fractions with the metabolites results in regulation of enzyme activities comparable to that seen in culture. This study examined whether changes in membrane fluidity might be one mechanism involved in the nongenomic regulatory pathway. Chondrocyte cultures were incubated with the vitamin D metabolites and changes in plasma membrane fluidity monitored using the fluorophore, TMA-DPH, which is specific for membranes exposed to external fluids. Isolated matrix vesicles were also incubated directly with the metabolites and anisotropy of the membrane, as well as alkaline phosphatase-specific activity, determined. 1,25-(OH)2D3 caused a rapid and constant increase in alkaline phosphatase-specific activity in GC matrix vesicles; 24,25-(OH)2D3 caused an increase in RC matrix vesicle enzyme activity that was both dose- and time-dependent. Matrix vesicles produced by GC had a lower degree of fluidity than their parent plasma membranes or RC plasma membranes and matrix vesicles. Fluidity of the GC membrane fractions was increased by 1,25-(OH)2D3 in a dose- and time-dependent manner. 1,25-(OH)2D3 had no effect on the fluidity of the RC membranes. 24,25-(OH)2D3 caused a decrease in fluidity in GC at later time points. This metabolite caused an increase in fluidity of RC plasma membranes that returned to normal levels by 6 h; however, the increase induced in the matrix vesicles remained elevated throughout the 24-h experimental period.(ABSTRACT TRUNCATED AT 250 WORDS)


Predictive modelling of the individual and combined effect of water activity and temperature on the radial growth of Fusarium verticilliodes and F. proliferatum on corn.
The major objective of this study was to develop validated models to describe the effect of a(w) and temperature on the radial growth on corn of the two major fumonisin producing Fusaria, namely Fusarium verticilliodes and F. proliferatum. The growth of these two isolates on corn was therefore studied at water activities between 0.810-0.985 and temperatures between 15 and 30 degrees C. Minimum a(w) for growth was 0.869 and 0.854 for F. verticilliodes and F. proliferatum, respectively. No growth took place at a(w) values equal to 0.831 and 0.838 for F. verticilliodes and F. proliferatum, respectively. The colony growth rates, g (mm d(-1)) were determined by fitting a flexible growth model describing the change in colony diameter (mm) with respect to time (days). Secondary models, relating the colony growth rate with a(w) or a(w) and temperature were developed. A third order polynomial equation and the linear Arrhenius-Davey model were used to describe the combined effect of temperature and a(w) on g. The combined modelling approaches, predicting g (mm d(-1)) at any a(w) and/or temperature were validated on independently collected data. All models proved to be good predictors of the growth rates of both isolates on maize within the experimental conditions. The third order polynomial equation had bias factors of 1.042 and 1.054 and accuracy factors of 1.128 and 1.380 for F. verticilliodes and F. proliferatum, respectively. The linear Arrhenius-Davey model had bias factors of 0.978 and 1.002 and accuracy factors of 1.098 and 1.122 for F. verticilliodes and F. proliferatum, respectively. The results confirm the general finding that a(w) has a greater influence on fungal growth than temperature. The developed models can be applied for the prevention of Fusarium growth on maize and the development of models that incorporate other factors important to mould growth on maize.


The proportion of intensive care unit admissions related to alcohol use: a prospective cohort study.
Alcohol abuse is a risk factor for serious illnesses, and a history of chronic alcohol abuse adversely affects the outcome of critically ill patients. It is not known what proportion of intensive care unit (ICU) admissions is related to alcohol use. Therefore, we investigated the proportion of emergency admissions related to alcohol. A prospective cohort study was conducted in a university hospital ICU. All adult patients (n = 893) who underwent emergency admission to our ICU during a period of 1 year were studied. The admitting physician determined whether there was a relationship between alcohol use and admission. ICU and hospital mortality and ICU length of stay (LOS) were recorded. The Therapeutic Intervention Scoring System (TISS) was used for ICU resource use estimation. There was a relationship between alcohol use and admission in 24% (215/893) of admissions and, in 156/893 admissions (17.5%), this seemed to be definite. ICU LOS was 1.2 days (0.7; 2.3) (median; interquartile range) for alcohol-related and 1.8 days (0.9; 3.6) for other admissions (P < 0.001). Patients with alcohol-related admissions consumed 17.8% of ICU patient-days and 18.7% of all accumulated TISS scores. ICU (8.8 vs. 10.5%, P = 0.603) and hospital (19.1 vs. 20.2%, P = 0.769) mortalities were no different between alcohol-related and other admissions. ICU admission is very often related to long-term chronic and/or occasional alcohol use.


Gα12/13 signaling in metabolic diseases.
As the key governors of diverse physiological processes, G protein-coupled receptors (GPCRs) have drawn attention as primary targets for several diseases, including diabetes and cardiovascular disease. Heterotrimeric G proteins converge signals from ~800 members of the GPCR family. Among the members of the G protein α family, the Gα12 family members comprising Gα12 and Gα13 have been referred to as gep oncogenes. Gα12/13 levels are altered in metabolic organs, including the liver and muscles, in metabolic diseases. The roles of Gα12/13 in metabolic diseases have been investigated. In this review, we highlight findings demonstrating Gα12/13 amplifying or dampening regulators of phenotype changes. We discuss the molecular basis of G protein biology in the context of posttranslational modifications to heterotrimeric G proteins and the cell signaling axis. We also highlight findings providing insights into the organ-specific, metabolic and pathological roles of G proteins in changes associated with specific cells, energy homeostasis, glucose metabolism, liver fibrosis and the immune and cardiovascular systems. This review summarizes the currently available knowledge on the importance of Gα12/13 in the physiology and pathogenesis of metabolic diseases, which is presented according to the basic understanding of their metabolic actions and underlying cellular and molecular bases.


Infectogenomics: insights from the host genome into infectious diseases.
Five years into the human postgenomic era, we are gaining considerable knowledge about host-pathogen interactions through host genomes. This "infectogenomics" approach should yield further insights into both diagnostic and therapeutic advances, as well as normal cellular function.


Dose-response effects of alcohol upon rat strains bred for differences in reactivity to alcohol.
Two rat strains, designated LA and MA, selectively bred for differential impairment of motor activity following an injection of alcohol, were tested in stabilmeters and compared over a range of ethanol doses. As expected, increasing doses of ethanol produced progressively greater activity decrements in both strains; however, the same dose of ethanol induced a more pronounced decrement in the MA strain than in LA strain at all doses. At the highest alcohol dose (2.25 g/kg), the LA animals were twice as active as were the MA strain at the 1.5 g/kg dose. This strain difference in impairment was evident within 3 min postinjection and remained throughout the 30 min test session. The results are discussed in terms of differential neural and behavioral toelrance to ethanol in the two strains.


Soil migration of antimony and arsenic facilitated by colloids in lysimeter studies.
The migration behaviors of antimony (Sb) and arsenic (As) and its influence factors have not been well understood among the different soils. In this study, we used lysimeter experiments to investigate the migration behavior of Sb compared with that of As in four representative soil materials from China. All the experiments processes and management measures were conducted to simulate the actual natural environmental conditions. Results indicated that after two years of leaching, the concentrations of Sb and As at the soil surface had decreased, whereas they increased in the deep soil profiles. In the polluted soil materials, 28.5%-39.2% of Sb and 0.4%-1.3% of As existed in the stable fraction, respectively. As and Sb levels were higher in the surface soil layer, and decreased with the soil depth in the different soil profiles. In soil leachate, Sb was mainly found in particle sizes smaller than 0.45 μm with the organic colloids, which had a peak in the spring and summer. On contrast, As was found in particle sizes larger than 0.45 μm with the inorganic colloids such as iron (Fe) and aluminum (Al) oxides. Pearson correlation results showed that the concentrations of Sb in the soil leaching solution and 0.45-μm-filltered solution were all positively correlated with Fe and Al. The results confirmed that Sb was combined with Fe and Al in the solution, and As posed a greater environmental risk than Sb during the leaching process. This study will help us to describe and predict As and Sb pollution in the soil environment, providing a basis for managing soil contaminated by these pollutants.


Broad-spectrum antibiotics associated gut microbiome disturbance impairs T cell immunity and promotes lung cancer metastasis: a retrospective study.
Gut microbiome has been linked to a regulatory role in cancer progression. However, whether broad-spectrum antibiotics (ATB) associated gut microbiome dysbiosis contributes to an impaired T cell immune function, and ultimately promotes lung cancer metastasis is not well known. In this study, a retrospective analysis was performed in a cohort of 263 patients initially diagnosed with non-small cell lung cancer (NSCLC) patients, including the ATB group (patients with broad-spectrum antibiotics treatment) (n = 124), and non-ATB group (n = 139) as control. ATB patients were prescribed ATB for over 5 days within 30 days prior to the collection of blood and fecal specimens and followed surgical treatment or first-line therapy. T cell immune function and metastasis-free survival (MFS) were evaluated between the two groups. Gut microbiota was evaluated by 16S rDNA sequencing. The predictive value of T cell immunity for MFS was evaluated by ROC analysis and Cox regression analysis. Our results suggest that broad-spectrum antibiotics (ATB) impair T cell immune function in patients with either early-stage or advanced NSCLC, which likely contribute to the promotion of lung cancer metastasis. Results of the survival analysis show that metastasis-free survival (MFS) is significantly shorter in the ATB patients than that in the non-ATB patients with stage III NSCLC. The 16S rDNA sequencing shows that ATB administration contributes to a significant dysbiosis of the composition and diversity of gut microbiota. Moreover, ROC analysis results of CD4 (AUC 0.642, p = 0.011), CD8 (AUC was 0.729, p < 0.001), CD16 + 56 + (AUC 0.643, p = 0.003), and the combination of CD4, CD8 and CD16 + 56+ (AUC 0.810, p < 0.001), or Cox regression analysis results of CD4 (HR 0.206, p < 0.001), CD8 (HR 0.555, p = 0.009), which is likely regulated by ATB administration, have significantly predictive values for MFS. These results provide evidence of gut microbiome disturbance due to ATB administration is involved in the regulation of T cell immunity, and their predictive value for the tumor metastasis in lung cancer patients. Thus, gut microbiota may serve as a therapeutic target for lung cancer. Consequently, caution should be exercised before the long-term administration of broad-spectrum antibiotics in cancer patients.


p300 (histone acetyltransferase) biomarker predicts prostate cancer biochemical recurrence and correlates with changes in epithelia nuclear size and shape.
p300 impacts the transcription of several genes involved in key pathways critical to PCa progression. Therefore, we evaluated the prognostic value of p300 expression and its correlation with nuclear alterations seen in tumor cells in men with long-term follow-up after radical prostatectomy (RP). NCI Cooperative Prostate Cancer Tissue Resource tissue microarray cores of 92 RP cases (56 non-recurrences and 36 PSA recurrences) were utilized for the study. p300 expression was assessed by quantitative immunohistochemistry and nuclear alterations in Feulgen-stained nuclei were evaluated by digital image analysis using the AutoCyte Pathology Workstation. Cox proportional hazards regression, Spearman's rank correlation, and Kaplan-Meier plots were employed to analyze the data. p300 expression significantly correlated with nuclear alterations seen in tumor cells; specifically with circular form factor (P = 0.012) and minimum feret (P = 0.048). p300 expression in high grade tumors (Gleason score >or=7) was significantly higher compared to low grade tumors (Gleason score <7) [17.7% versus 13.7%, respectively, P = 0.03]. TNM stage, Gleason score, and p300 expression were univariately significant in the prediction of PCa biochemical recurrence-free survival (P <or= 0.05). p300 expression remained significant in the multivariate model (P = 0.03) while Gleason score showed a trend toward significance (P = 0.06). Patients with a Gleason score >or=7 and p300 expression >24% showed the highest risk for PCa biochemical recurrence (P = 0.002). p300 expression correlates with nuclear alterations seen in tumor cells and has prognostic value in predicting long-term PCa biochemical recurrence-free survival.


[Internal injuries of larynx in children. Case reports].
The authors present larynx injuries among children especially with regard to iatrogenic internal injuries based on personally observed cases: dislocation of arytenoid cartilage in 11 year-old girl and post intubation granuloma in 8 year-old girl.


Surgical site infections after elective craniotomy for brain tumor: a study on potential risk factors and related treatments.
Surgical site infection (SSI) is a common complication following craniotomy that increases morbidity, mortality, and medical expenses. The objectives of this study were to determine the relevant risk factors associated with SSI after elective craniotomy for brain tumor and analyse the treatments for SSI. A retrospective nested case‒control study was conducted using data from patients who underwent craniotomy for brain tumor resection at the Neurosurgical Oncology Department No. 6 of Beijing Tiantan Hospital, Capital Medical University, between January 2019 and December 2021. Risk factors for SSI were determined using multivariate logistic regression analysis. We analyzed microbiological and related treatment data for different SSI types. Among 2061 patients who underwent craniotomy for brain tumor, 31 had SSI (1.50%). In the multivariate logistic regression analysis, body mass index (BMI) and operative duration were identified as independent risk factors for SSI. The most common microorganism isolated from SSIs was Staphylococcus epidermidis (22.9%), and drug sensitivity results showed that gram-positive bacteria were sensitive to linezolid, vancomycin and tigecycline, whereas gram-negative bacteria were sensitive to meropenem, cefepime and ceftazidime. Six of the seven patients who underwent bone flap removal due to osteomyelitis were infected with gram-negative bacteria. BMI and operative duration were identified as independent risk factors for SSI. Diabetes mellitus, previous ratio therapy, type of incision, recurrence tumor and other risk factors were not found to be associated with the occurrence of SSI in this study.


Gonadotropin suppression with oral contraceptives before in vitro fertilization.
One hundred eighty-one stimulation cycles in which gonadotropin suppression with oral contraceptives (OCs) preceded induction of follicle stimulation (study group) and 113 stimulation cycles without pituitary suppression (control group) were compared. The mean length of ovarian suppression was 35.3 +/- 0.9 days. No spontaneous luteinizing hormone (LH) surges occurred when the use of OC preceded ovarian hyperstimulation, whereas in the control group the incidence of LH surges was 19.5%. The mean amount of human menopausal gonadotropin required was significantly lower in the study group than in the control group (8.9 +/- 0.4 and 10.9 +/- 0.4 ampules, respectively). Significantly more follicles greater than or equal to 1.5 cm in diameter were seen on the day before oocyte retrieval and significantly more oocytes were retrieved per attempt in the group with OC pretreatment. Our data clearly demonstrate that OCs are useful in vitro fertilization stimulation protocols to facilitate scheduling of cycles and to prevent spontaneous LH surges.


In vitro and in vivo microdialysis calibration for the measurement of carbamazepine and its metabolites in rat brain tissue using the internal reference technique.
Microdialysis, as in vivo sampling technique, can be used for determining exogenous compounds in the extracellular fluid of freely moving animals and in humans. Usually, calibration of the microdialysis probe is determined by in vitro relative recovery (RR) (dialysate extraction fraction). However, due to different diffusion properties of the compound in tissue, the RR in vivo is different from the RR in vitro. In this study, the evaluation of the internal reference technique as in vivo calibration method was established. To determine the RR in vivo, the relative loss (RL) was defined as the loss of a compound from the perfusate. RL was determined in vitro and in vivo by adding an internal standard (IS) to the perfusate. This internal reference technique was applied for the determination of carbamazepine (CBZ) and its 2 major metabolites, carbamazepine-10,11-epoxide (CBZ-EPO) and trans-10,11-dihydroxy-10,11-dihydro-carbamazepine (CBZ-DIOL) using 2-methyl-5H-dibenz(b,f)azepine-5-carboxamide (m-CBZ) as IS. In vitro and in vivo, the loss of m-CBZ and the recovery of CBZ are identical. The ratios of the RR of CBZ-EPO and CBZ-DIOL to the RL of m-CBZ are constant, in vitro and in vivo. Therefore, m-CBZ can be used as IS for CBZ, CBZ-EPO and CBZ-DIOL determinations in brain tissue. It is shown that the internal reference technique is a useful method to estimate the true concentration of exogenous compounds in the extracellular space of tissues.


Bis(5-hydroxy-2-hydroxymethyl-pyran-4-one-6-yl)methane: a novel ligand for the intracellular mobilisation of ferritin-bound iron.
The efficacy of a novel tetradentate iron(III) ligand for the in vitro mobilisation of ferritin-bound iron is measured in direct comparison to the clinically approved agents, 1,2-dimethyl-3-hydroxypyridin-4-one and desferrioxamine.


Asparagine-84, a regulatory allosteric site residue, helps maintain the quaternary structure of Campylobacter jejuni dihydrodipicolinate synthase.
Dihydrodipicolinate synthase (DHDPS) from Campylobacter jejuni is a natively homotetrameric enzyme that catalyzes the first unique reaction of (S)-lysine biosynthesis and is feedback-regulated by lysine through binding to an allosteric site. High-resolution structures of the DHDPS-lysine complex have revealed significant insights into the binding events. One key asparagine residue, N84, makes hydrogen bonds with both the carboxyl and the α-amino group of the bound lysine. We generated two mutants, N84A and N84D, to study the effects of these changes on the allosteric site properties. However, under normal assay conditions, N84A displayed notably lower catalytic activity, and N84D showed no activity. Here we show that these mutations disrupt the quaternary structure of DHDPS in a concentration-dependent fashion, as demonstrated by size-exclusion chromatography, multi-angle light scattering, dynamic light scattering, small-angle X-ray scattering (SAXS) and high-resolution protein crystallography.


Inclusion of babassu bran produced in milk production in Amazonia.
One important species for extraction and subsistence of traditional populations in Brazil is the babassu palm (Attalea speciosa Mart ex Spreng). As a by-product of the babassu oil socio-productive chain in Amazonia region, the residual bran needs to be better used and could represent an additional source of income for coconut breakers, and a local alternative in the composition of diets for dairy cows. The objective of this work was to evaluate the effect of a diet containing babassu bran on the intake and digestibility of cows, their production and milk composition, in the Amazon region. The experimental trial was arranged in a Cross over design, composed of fourteen animals, two treatments and two periods of 28 days (21 days for adaptation to diets and 7 days for data collection). Treatments were concentrate without babassu bran (CNV) and with babassu bran (BBS). Milk composition did not differ significantly between treatments. The inclusion of 35.6% BBS in feed ration supported an average daily milk production of 18.2 kg/day, while CNV produced 19.8 kg/day. There was lower feed consumption for BBC, but equal to that of CNV when comparing liters of milk produced per kg of ingested concentrate (1:3.09 CNV × 1:3.07 BBS), which demonstrates that treatments did not differ in productive efficiency. Our results bring the possibility of a better use of babassu bran. This alternative could improve the quality of life of rural populations, while being linked to the conservation of native ecosystems and the sustainable use of socio-biodiversity products.


Treatment of malaria: some considerations and limitations of the current methods of assessment.
The currently used methods for assessing the therapeutic response to antimalarial drugs are relatively imprecise and insensitive. These methods are inadequate in severe malaria when the objectives of treatment are to save life and prevent complications. Very large studies are needed to demonstrate significant differences in mortality, but measurement of the rates of clinical, biochemical, and parasitological response may provide useful comparative information. Definitions, assessment criteria, procedures, and data collection forms should be standardized and evaluated prospectively. Antimalarial drug treatment in different clinical situations should be assessed in terms of the balance between the risks of drug toxicity and the benefits of the antimalarial drug action. This balance is considerably different in severe falciparum malaria compared with uncomplicated malaria infections.


Quantitative assessment of HIV-1 protease inhibitor interactions with drug efflux transporters in the blood-brain barrier.
To quantitatively characterize the drug efflux interactions of various HIV-1 protease inhibitors in an in vitro model of the blood-brain barrier (BBB) and to compare that with HIV-1 protease inhibitor stimulated P-glycoprotein (P-gp)-ATPase activity. Cellular accumulation of the P-gp sensitive probe, rhodamine 123 (R123), and the mixed P-gp/multidrug resistance-associated protein (MRP) probe, 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF), were evaluated in primary cultured bovine brain microvessel endothelial cells (BBMEC) in the presence of various concentrations of HIV-1 protease inhibitors. The potency (IC50) and efficacy (Imax) of the drugs in the cell accumulation assays for P-gp and/or MRP was determined and compared to activity in a P-gp ATPase assay. For R123 (P-gp probe), the rank order potency for inhibiting R123 accumulation in the BBMEC was saquinavir=nelfinavir>ritonavir=amprenavir>indinavir. This correlated well with the rank order affinity in the P-gp ATPase assay. The rank order potency for MRP-related drug efflux transporters, was nelfinavir>ritonavir>saquinavir>amprenavir>indinavir. HIV-1 protease inhibitors potently interact with both P-gp and MRP-related transporters in BBMEC. Characterization of the interactions between the HIV-1 protease inhibitors and drug efflux transporters in brain microvessel endothelial cells will provide insight into potential drug-drug interactions and permeability issues in the BBB.


Hypothermia-induced potentiation of histamine H2-receptor-mediated relaxation and cyclic AMP increase in the isolated mesenteric artery of the rabbit.
On helically cut strips of the rabbit's mesenteric artery, a temperature decrease from 42 degrees C to 25 degrees C reduced the contractile responses to histamine. Metiamide shifted the dose-response curve of the histamine-induced contraction towards higher values at 25 degrees C, but not at 42 degrees C. Furthermore, on arterial strips contracted by phenylephrine histamine evoked a dose-dependent relaxation at 25 degrees C whereas at 42 degrees C only slight relaxing responses to histamine occurred. Metiamide was capable of preventing the relaxation induced by histamine in a competitive manner. At 25 degrees C the relaxation as produced by histamine was accompanied by increases in cyclic AMP which occurred prior to the relaxing effects. Metiamide abolished the cyclic AMP increase in response to histamine. At 42 degrees C histamine was unable to elevate the cyclic AMP content. Thus, it is concluded that a cyclic AMP-mediated relaxation due to stimulation of H2-receptors counteracts the histamine-induced contraction and reduces the contractile responses to histamine at low temperatures. In addition, clear-cut evidence exists from the present study that also on artery smooth muscle the H2-receptor-mediated responses are closely associated to cyclic AMP.


Community engagement processes in low- and middle-income countries health research settings: a systematic review of the literature.
Community Engagement is an important ethical imperative in research. Although substantial research emphasizes its real value and strategic importance, much of the available literature focuses primarily on the success of community participation, with little emphasis given to specific community engagement processes, mechanisms and strategies in relation to intended outcomes in research environments. The systematic literature review's objective was to explore the nature of community engagement processes, strategies and approaches in health research settings in low- and middle-income countries. The systematic literature review design was informed by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. We searched for peer-reviewed, English-language literature published between January 2011 and December 2021 through three databases on the internet (PubMed, Web of Science and Google Scholar). The terms "community engagement," "community involvement," "participation," "research settings," and "low- and middle-income countries" were merged in the search. The majority of publications [8/10] were led by authors from low- and middle-income countries, with many of them, [9/10] failing to continuously include important aspects of study quality. Even though consultation and information sessions were less participatory, articles were most likely to describe community engagement in these types of events. The articles covered a wide range of health issues, but the majority were concerned with infectious diseases such as malaria, human immunodeficiency virus, and tuberculosis, followed by studies on the environment and broader health factors. Articles were largely under-theorized. Despite the lack of theoretical underpinnings for various community engagement processes, strategies and approaches, community engagement in research settings was variable. Future studies should go deeper into community engagement theory, acknowledge the power dynamics underpin community engagement, and be more practical about the extent to which communities may participate.


Forecasting of the wind speed under uncertainty.
In this paper, the semi-average method under neutrosophic statistics is introduced. The trend regression line for the semi-average method is given in the presence of Neutrosophy in the data. The application of the semi-average method under indeterminacy is given with the help of wind speed data. The efficiency of the semi-average method under the neutrosophic statistics is discussed over the semi-average method under classical statistics. From the analysis, it is concluded that the proposed method is effective, informative, and flexible for the forecasting of wind speed.


The lipid-water interface: revelations by osmotic stress.
Lipids at the bilayer-water interface are highly disordered and mobile, and large areas of the bilayer undergo thermal undulations. Water penetrates significantly down to the hydrocarbon chain level. This water, and water out to about 10 A from the surface, is perturbed by the lipid surface in a way that produces a strong hydration repulsion and precludes molecular contact between bilayers. Its removal costs work, but most of this water is neither a permeable barrier nor unavailable to solvate other solutes. All hydrophilic surfaces show this "hydration force." Most lipids have an excess higher free energy when packed within a bilayer membrane since in isolation they pack into high curvature assemblies with polar groups on the concave side. Osmotic stress measurements of those curved assemblies yield a measure of monolayer elastic parameters and the excess higher free energy, which likely controls embedded proteins. Osmotic stress experiments can determine whether water is energetically significant, or not, in almost any system. The osmotic effect of solutes, independent of specific binding, is to compete with lipids and proteins for water. Solute affinity for water can modify lipid packing and protein conformation, coupling lipid and protein structure and function to osmolality at the molecular level.


Development of LRRK2 Inhibitors for the Treatment of Parkinson's Disease.
Linkage and genome-wide association studies have identified a genetic risk locus for late-onset Parkinson's disease in chromosome 12, originally identified as PARK6. The causative gene was identified to code for a large multifunctional protein, LRRK2 (leucine-rich repeat kinase 2). The combined genetic and biochemical evidence supports a hypothesis in which the LRRK2 kinase function is causally involved in the pathogenesis of sporadic and familial forms of PD, and therefore that LRRK2 kinase inhibitors could be useful for treatment. Although LRRK2 has so far not been crystallised, the use of homology modelling and crystallographic surrogates has allowed the optimisation of chemical structures such that compounds of high selectivity with good brain penetration and appropriate pharmacokinetic properties are now available for understanding the biology of LRRK2 in vitro and in vivo. This chapter reviews LRRK2 biology, the structural biology of LRRK2 and gives an overview of inhibitors of LRRK2.


The mitochondria/caspase-dependent apoptotic pathway plays a role in the positive effects of a power frequency electromagnetic field on Alzheimer's disease neuronal model.
In this study, rat pheochromocytoma (PC12) cells were induced into an Alzheimer's Disease (AD) neuronal model using nerve growth factor (NGF; 50 ng/mL) and Amyloid β25-35 (20 μmol/L). Changes in the morphological structure, cell viability, apoptosis rate, and expression of apoptosis-related protein induced by exposure to a power frequency electromagnetic field (PF-MF; 50 Hz, 100 μT, 24 h) were detected respectively by light and electron microscopy, the MTT assay, immunohistochemistry, flow cytometry and enzyme-linked immunosorbent assays. The results showed that 3-12 h after PF-MF exposure, the pathological injury was improved partly; metabolic activity was promoted and cell apoptosis was inhibited in the AD neuronal model. In addition, PF-MF exposure significantly inhibited the expression of Caspase8, Caspase3, and CytC, but increased the Bcl-2/Bax ratio of the AD neuronal model. Meanwhile, PF-MF seemed to have no effect on the expression of Fas and TNFR1. This study indicated that the mitochondria/caspase-dependent apoptotic pathway plays an important role in the positive effects of PF-MF on an AD neuronal model. The results suggested that PF-MF exposure might have potential therapeutic value for AD, and the underling molecular mechanisms still need further studies.


Effects of limited enzymatic hydrolysis and polysaccharide addition on the physicochemical properties of emulsions stabilized with duck myofibrillar protein under low-salt conditions.
This study aimed to investigate the role of hydrolysis and guar gum (GG) participation on the emulsification of the duck myofibrillar protein (MP) and the related stability of oil-in-water emulsion in low-salt condition. Emulsions were prepared using one of each or both treatments, and that prepared with trypsin hydrolysis and GG (T-GG) exhibited the highest stability. FTIR analysis confirmed the hydrogen bond interactions between the system components. T-GG treatment improved emulsion properties and decreased oil droplet size. Moreover, CLSM indicated that aggregation of T-GG oil droplets was prevented. Physical stability was assessed such as Turbiscan stability index, creaming index, and rheological properties. The adsorbed percentage for T-GG was the lowest. However, interfacial tension, droplet size, stability, and peroxide value analyses indicated that a denser interfacial membrane structure is formed with T-GG. Thus, T-GG treatment could be applied in the food industry, such as in nutrient delivery systems and fat mimetics.


Assessment of risk factors for emergence distress and postoperative behavioural changes in children following general anaesthesia.
Emergence distress commonly occurs in children recovering from the immediate effects of general anaesthesia. This study was performed to (1) examine whether parental presence in the operating room during emergence from anaesthesia reduces the incidence or severity of emergence distress behaviour, and (2) assess psychosocial risk factors, including child temperament and sleep behaviour, for development of emergence distress. A randomized and controlled trial of parental presence at emergence was conducted in 100 ASA class I and II children having general anaesthesia for inguinal or penile surgery. Children in the study group had a parent present at induction and emergence of anaesthesia, while children in the control group had a parent present only at induction. Emergence and postanaesthesia care unit (PACU) behaviour was monitored using both the Operating Room Behaviour Rating Scale (ORBRS) and a 7-point Likert type cooperation scale. One-way anovas showed no significant differences between the control group and the study group on emergence distress behaviour. The frequency of negative postoperative behavioural changes at 1 and 4 weeks postsurgery was low in both groups. Children described as clingy/dependent (chi2 = 5.57, P < 0.06) and children with frequent temper tantrums (chi2 = 7.44, P < 0.02) were more likely to have emergence distress behaviour. Parental presence during emergence from anesthesia did not decrease the incidence or severity of emergence distress behaviour in children. Young children and children with a history of temper tantrums or separation anxiety may be more likely to develop such behaviour.


Oncocytic neoplasms of salivary glands: an ultrastructural study.
The oncocyte is a peculiar cell found in the aging parotid gland. A precise definition of oncocytic tumors in now available using histochemical and electron microscopic techinques. Oncocytic carcinomas are very rare; only a handful have been cited in the literature. This is the first description of the electron microscope features of oncocytic carcinomas we can find in the literature. The cells of these carcinomas are characterized by mitochondrial hyperplasia, pleomorphism and atypism. The cells contained few other subcellular organelles and no myofilaments or glycogen as previously described in benign oncocytomas. In addition to these electron microscopic features, the clinical presentation, treatment, and biologic course of these three cases of oncocytic carcinoma and two cases of benign oncocytoma will be presented.


Activated sludge microbiome with H2O2-modified biochar enhances the treatment resilience and detoxification of oxytetracycline and its toxic byproducts.
The widespread presence of oxytetracycline (OTC) in aquatic ecosystems poses both health risks and ecological concerns. The present study revealed the beneficial role of hydrogen peroxide (H2O2)-pretreated biochar (BC) derived from agricultural hardwood waste in an activated sludge (AS) bioprocess. The BC addition significantly enhanced the removal and detoxification of OTC and its byproducts. BC was initially modified using H2O2 to improve its OTC adsorption. Two AS reactors were then established, one with H2O2-modified BC and one without, and both were exposed to OTC. The BC-added reactor exhibited significantly higher OTC removal rates during both the start-up (0.97 d-1) and steady-state (0.98 d-1) phases than the reactor without BC (0.54 d-1 and 0.83 d-1, respectively). Two novel transformation pathways for OTC were proposed, with four byproducts originating from OTC identified, some of which were found to be more toxic than OTC itself. The BC-added reactor had significantly higher system functioning in terms of its heterotrophic activity and the reduction of the toxicity of OTC and its byproducts, as illustrated by structure-based toxicity simulations, antimicrobial susceptibility experiments, analytical chemistry, and bioinformatics analysis. Bioinformatics revealed two novel bacterial populations closely related to the known OTC-degrader Pandoraea. The ecophysiology and selective enrichment of these populations suggested their role in the enzymatic breakdown and detoxification of OTC (e.g., via demethylation and hydrogenation). Overall, the present study highlighted the beneficial role of H2O2-modified BC in combination with the AS microbiome in terms of enhancing treatment performance and resilience, reducing the toxicological disruption to biodiversity, and detoxifying micropollutants.


Efficacy and safety of allergen immunotherapy in children.
To determine the efficacy and safety of immunotherapy in children with allergic rhinitis and allergic asthma. Current Contents database for 1967 to 2005. The expert opinion of the author was used to select studies for inclusion in this review. Meta-analyses have confirmed the clinical effectiveness of allergen immunotherapy in patients with allergic rhinitis and asthma; however, most studies involved immunotherapy with a single allergen. Special considerations of safety regarding immunotherapy in children are also necessary, since fatalities have been reported. Fatal reactions occur primarily in patients with asthma and particularly those whose asthma is not well controlled. There is probably increased risk during the build-up phase and with the first injection from a new vial. Injections at home or in clinics without adequate supervision also constitute an increased risk of a fatal outcome. Specific immunotherapy has been demonstrated to have some protective effect against the development of additional sensitivities in the monosensitized child and to reduce the risk of developing asthma in children with allergic rhinitis. Because of the modification of the underlying immune process, the beneficial effects of immunotherapy persist for a long but not yet fully determined period after the cessation of treatment. This persisting effect makes immunotherapy an attractive supplement to symptomatic treatment of children with allergic asthma and allergic rhinitis. The treatment is not without risk, however, especially in the child with asthma, so it should be administered with caution.


Progressive hypoxia inhibits the de novo synthesis of galactosylceramide in cultured oligodendrocytes.
Neonatal rat oligodendrocyte (OLG) cultures exposed to 6 h of gradual, progressive hypoxia in a GasPak (BBL, Becton Dickinson) apparatus were not injured or metabolically impaired, but instead showed a specific inhibition of de novo synthesis (measured by [3H]palmitic acid labeling) of the major myelin component galactosylceramide (GalCer). De novo synthesis of the 2-hydroxy fatty acid GalCer (HFA-GalCer) species, which requires O2 for its synthesis, was most severely inhibited (by 65%), while non-hydroxy GalCer species (NFA-GalCer) were less affected. The synthesis of membrane glycerophospholipids and sphingomyelin was unaffected by hypoxia. Treatment of OLG with 12 nM oligomycin, an inhibitor of mitochondrial ATP synthesis, resulted in an inhibition (by 50-60%) of synthesis of all GalCer species. [3H]Palmitate labeling of NFA-ceramide, the ungalactosylated precursor of NFA-GalCer species, increased in both hypoxia and oligomycin treatments, suggesting that the conversion of newly synthesized ceramide to GalCer was blocked. Newly synthesized HFA-ceramide did not accumulate in OLG, but the small labeled HFA-ceramide pool present during hypoxia was not converted into HFA-GalCer. Pulse-chase studies indicated that NFA- and HFA-ceramides labeled during these treatments were available for galactosylation and could be converted into GalCer upon reoxygenation. [3H]Galactose labeling of NFA-GalCer species was enhanced 2-fold in hypoxia, in contrast to the inhibition seen with [3H]palmitic acid labeling. Thus, while de novo GalCer synthesis was blocked in hypoxia, galactosylation of pre-existing ceramide pools was actually enhanced. Our evidence suggests that hypoxia results in a reversible inhibition of transport of newly synthesized ceramide from its site of synthesis to its site of galactosylation, but causes an increase in galactosylation of subcellular pools of pre-existing ceramide.


Evaluation of knowledge, attitudes, and practices of deer owners following identification of a cluster of captive deer with rabies in Pennsylvania in July 2010.
To evaluate knowledge, attitudes, and practices of deer owners following identification of a cluster of captive deer with rabies as an aid for the development of rabies prevention educational materials. Cross-sectional study. Captive-deer owners who were members of the pennsylvania deer farmers association. Information was obtained via a mailed, self-administered questionnaire. The questionnaire response rate was 59% (249/425). One hundred three of 206 (50%) respondents had incomplete knowledge of rabies virus vectors, transmission, severity, and prevention measures. Birds or snakes were incorrectly identified as rabies vectors by 96 of 213 (45%) respondents, and most (≥ 94%) respondents identified rabies virus reservoirs as vectors. Ninety of 231 (39%) respondents identified death as an outcome of rabies, and 184 of 235 (78%) respondents would seek emergency treatment if they suspected exposure. Only 62 of 235 (26%) respondents would wash a wound immediately. The majority of respondents (173/239 [72%]) did not know the clinical signs of rabies in deer. Nine respondents indicated that they vaccinated their deer against rabies, and the majority of respondents (158/214 [74%]) would be willing to vaccinate. Findings suggested that deer owners in Pennsylvania have a basic knowledge of rabies; however, knowledge, attitudes, and practices regarding prevention of rabies transmission could be improved considerably. Rabies educational materials for deer owners should focus on postexposure procedures, disease severity, recognition of rabies in deer, and changes in management practices such as vaccination to prevent rabies.


Bilateral localized amyloidosis of the ureter presenting with anuria.
A patient with bilateral localized amyloidosis of the ureters is described. This is the first case to be reported in which anuria was the presenting symptom, the second case with bilateral involvement and the thirteenth case of amyloidosis localized to the ureter. Careful urologic and hematologic followup is indicated.


Sulfur-containing constituents and one 1H-pyrrole-2-carboxylic acid derivative from pineapple [Ananas comosus (L.) Merr.] fruit.
Two sulfur-containing compounds, (S)-2-amino-5-((R)-1-carboxy-2-((E)-3-(4-hydroxy-3-methoxyphenyl)allylthio)ethyl-amino)-5-oxopentanoic acid (1) and (S)-2-amino-5-((R)-1-(carboxymethylamino)-3-((E)-3-(4-hydroxyphenyl)allylthio)-1-oxopropan-2-ylamino)-5-oxopentanoic acid (2), and one 1H-pyrrole-2-carboxylic acid derivative, 6-(3-(1H-pyrrole-2-carbonyloxy)-2-hydroxypropoxy)-3,4,5-trihydroxy-tetrahydro-2H-pyran-2-carboxylic acid (3), together with eighteen known phenolic compounds, were isolated from the fruits of pineapple. Their structures were elucidated by a combination of spectroscopic analyses. Some of these compounds showed inhibitory activities against tyrosinase. The half maximal inhibitory concentration values of compounds 1, 4, 5, 6, 7 are lower than 1 mM. These compounds may contribute to the well-known anti-browning effect of pineapple juice and be potential skin whitening agents in cosmetic applications.


Increased radioresistance of an in vitro transformed human small cell lung cancer cell line.
After 4-6 months in continuous culture the human small cell lung cancer (SCLC) cell line, U-1906, changed its radiobiological characteristics spontaneously. The cell line became more radioresistant indicating an increased repair capacity. This change was accompanied by a more adherent growth pattern, a higher clonogeneity, a decrease in the cytokeratin (tissue polypeptide antigen) content and increased glucagon and neuron-specific enolase (NSE) production. Other parameters such as the estramustine-binding protein (EMBP) and the proliferation associated antigen Ki-67 were unaltered. This spontaneous transformation in vitro of U-1906 may reflect a clinically important in vivo phenomenon of SCLC, which frequently develops resistance both to radio- and chemotherapy.


Building a dual-energy CT service line in abdominal radiology.
As the access of radiology practices to dual-energy CT (DECT) has increased worldwide, seamless integration into clinical workflows and optimized use of this technology are desirable. In this article, we provide basic concepts of commercially available DECT hardware implementations, discuss financial and logistical aspects, provide tips for protocol building and image routing strategies, and review radiation dose considerations to establish a DECT service line in abdominal imaging. KEY POINTS: • Tube-based and detector-based DECT implementations with varying features and strengths are available on the imaging market. • Thorough assessment of financial and logistical aspects is key to successful implementation of a DECT service line. • Optimized protocol building and image routing strategies are of critical importance for effective use and seamless inception of DECT in routine clinical workflows.


An experimental study on antitumoral effects of KI-21-3, a synthetic fragment of antimicrobial peptide LL-37, on oral squamous cell carcinoma.
The aim of this study was to investigate the oncolytic properties of KI-21-3, a shortened fragment of LL-37, against oral squamous cell carcinoma (OSCC) in an animal model. Twelve athymic nude mice were divided into a therapy and a control group of six animals each. In both groups, SCC-4 cells were administered extraorally into the floor of the mouth in order to create an OSCC model. In the study group, KI-21-3 was applied intravenously during the 8th and 9th weeks. The subjects in the control group were injected with phosphate buffered saline solution in the same manner. During an examination period of 12 weeks, weight control was performed twice a week. Tumor growth was further controlled volumetrically via ultrasonography once a week with regular intervals. Following sacrifice, ablated tumoral tissues were immunohistochemically evaluated in order to determine the proliferation and apoptotic properties. The mean tumor weight in the AMP group was 0.0236 ± 0.023 g, which was 30% lower than the control group with the mean value of 0.01651 ± 0.012 g. In the control group, the approximate number of the proliferating cells per visualized field was fourfold higher compared to the therapy group. Moreover, in the control group, the number of apoptotic cells per visualized field was significantly lower compared to the therapy group. KI-21-3 showed considerable oncolytic properties on SCC-4 carcinoma cells via antiproliferative and caspase-3 apoptotic pathway. Further investigations are necessary to clarify the dose-dependent effects of this agent.


Hypofractionated external radiation with high and low dose rates in the treatment of advanced cancer of the cervix.
Thirteen patients with cancer of the cervix Stage IIIb (heavy) were treated with high (9) and low (4) dose rates, once a week. A total dose of 4200 cGy was delivered over 7 weeks with a fraction size of 600 cGy. Six of nine patients showed complete response following high dose rate hypofractionated radiation. It is felt that synchronization of cells into a sensitive phase during low dose rate radiation is not the determinant of outcome even when duration of treatment is only 5 to 6 hours.


Patency of heparin-PVA coated tubes at low flow rates.
The patency of heparin-PVA coated 1.14 mm i.d. polyethylene tubes was greater than that of control tubes without heparin in a novel test section that enabled ex vivo evaluation in dogs at low flow and shear rates (2-10 ml/min). Low shear rates were achieved by diverting a small portion of a chronic A-V shunt flow through Y-connectors into the small diameter test tubing. For shear rates in the range of 200-650 s-1 half the heparin-PVA tubes remained patent for longer than 72 min whereas half the control tubes (PVA only) only remained patent for longer than 30 min. At higher shear rates (500-1000 s-1), the 50% patency times for heparin-PVA and PVA were 225 and 123 min respectively. The increased patency at the higher shear rates was attributed more to the effect of the connector than just shear rate. The higher shear rates were achieved by changing from a Y-connector in which the small diameter side-branch was cut flush to the larger diameter main line to one in which the small diameter test tubing protruded into the lumen of the main line; the latter 'protruding' connector was presumed to result in less platelet pre-activation than the other. This method has been found useful for assessing the thrombogenicity of heparinized tubes at low flow rates.


Metal Content in Caps and Stalks of Edible Mushrooms: Health Benefits and Risk Evaluation.
Mushrooms are a good source of protein and phenolic compounds which provides health benefits for humans. The purpose of this study was to compare the content of eight metals, protein, and total phenolics (TPC) of 5 different species (Agaricus bisporus-white and brown mushrooms, Agaricus cupreobrunneus, Auricularia cornea, Hypsizgus tesselatus, and Pleurotus eryngii species-complex) of edible mushrooms available on the Romanian market. Agaricus bisporus and Agaricus cupreobrunneus were purchased and cultivated in Romania and the other species were cultivated in other countries (Turkey and China). The metal content determined by graphite atomic absorption spectrometry (GTAAS) varied in the order Cu > Pb > Ni > Fe > Cr > Mn > Co > Cd. Almost all the samples contained a greater quantity of metals in the stalk than in the cap. In addition, the levels of toxic metals were low. The protein content of analyzed samples ranged from 0.0926 to 0.2743%, the highest value being observed in Pleurotus eryngii species-complex mushroom. TPC of extracts increased over time but there was a variability in the concentration for each mushroom species (0.25-12.25 mg GAE/g). The investigated mushroom species possess no health risk and may be potential nutritional supplements for human diets due to their phenolic compounds, protein, and mineral content.


LncRNA LINC00961 regulates endothelial‑mesenchymal transition via the PTEN‑PI3K‑AKT pathway.
The long noncoding RNA LINC00961 plays a crucial role in cancer and cardiovascular diseases. In the present study, the role and underlying mechanism of LINC00961 in endothelial‑mesenchymal transition (EndMT) induced by transforming growth factor beta (TGF‑β), was investigated. Human cardiac microvascular endothelial cells were transfected with LV‑LINC00961 or short hairpin LINC00961 plasmids to overexpress or knock down LINC00961 in the cells, respectively. The cells were then exposed to TGF‑β in serum‑free medium for 48 h to induce EndMT. Flow cytometric analysis, Cell Counting Kit‑8 assay and immunofluorescence staining were performed to examine the cell apoptosis rate, assess cell viability, and identify CD31+/α‑SMA+ double‑positive cells, respectively. Western blotting and reverse transcription‑ quantitative polymerase chain reaction were used to evaluate protein and mRNA expression, respectively. Injury to endothelial cells and EndMT was induced by TGF‑β in a time‑dependent manner. LINC00961 overexpression promoted injury and EndMT, whereas LINC00961 knockdown had the opposite effects. Knockdown of LINC00961 attenuated EndMT and injury to endothelial cells induced by TGF‑β via the PTEN‑PI3K‑AKT pathway. Inhibition of LINC00961 expression may prevent the occurrence of EndMT‑related cardiovascular diseases, such as myocardial fibrosis and heart failure. Therefore, LINC00961 shows potential as a therapeutic target for cardiovascular diseases.


Pre-operative serum inflammation-based scores in patients with pituitary adenomas.
Full blood count (FBC) and serum inflammation-based scores reflect systemic inflammation and predict outcomes in cancer, but little is known in pituitary adenomas (PAs). We aimed to characterise FBC and inflammation-based scores in PA patients and investigate their usefulness in predicting challenging disease course. We studied 424 PA patients first operated at our centre with available pre-operative biochemical data. Patients with infection, malignancies, autoimmune or haematological conditions were excluded. Inflammation-based scores studied: Neutrophil-to-Lymphocyte Ratio (NLR), Platelet-to-Lymphocyte Ratio (PLR), Lymphocyte-to-Monocyte Ratio (LMR), Systemic Immune-Inflammation Index (SII), Neutrophil-Platelet Score (NPS), Prognostic Nutrition Index (PNI), and Glasgow Prognostic Score (GPS). Cushing's disease patients had more platelets, leucocytes, neutrophils and monocytes, and higher NLR, NPS and SII. Serum inflammation-based scores didn't differ among non-Cushing PA subtypes. The glucocorticoid excess severity influenced leucocyte, eosinophil, basophil and platelet counts, and GPS in Cushing's disease. Patients with functioning non-Cushing PAs with suprasellar extension, cavernous sinus invasion and hypopituitarism had GPS ≥ 1, while NPS ≥ 1 was associated with suprasellar extension. More invasive and difficult to treat corticotrophinomas were associated with fewer platelets pre-operatively (< 299.5 × 109/L predicting multimodal treatment). Non-functioning PA patients who suffered apoplexy had more leucocytes, neutrophils and monocytes, higher GPS ≥ 1 and fewer platelets; re-operated cases had fewer lymphocytes, higher NLR and PLR. Serum inflammation-based scores may predict invasive/refractory PAs: GPS and PNI in non-functioning and functioning non-Cushing PAs; NPS in functioning non-Cushing PAs; NLR and PLR in non-functioning PAs. Platelets < 299.5 × 109/L predict multimodal treatment in Cushing's disease. Further studies are needed to confirm these observations.


Comparative Genomics and Evolutionary Modularity of Prokaryotes.
The soaring number of high-quality genomic sequences has ushered in the era of post-genomic research where our understanding of organisms has dramatically shifted towards defining the function of genes within their larger biological contexts. As a result, novel high-throughput experimental technologies are being increasingly employed to uncover physical and functional associations of genes and proteins in complex biological processes. Through the construction and analysis of physical, genetic and metabolic networks generated for the model organisms, such as Escherichia coli, organizational principles of the genome have been deduced, such as modularity, which has important implications toward understanding prokaryotic evolution and adaptation to novel lifestyles.


Working Memory Impairing Actions of Corticotropin-Releasing Factor (CRF) Neurotransmission in the Prefrontal Cortex.
The prefrontal cortex (PFC) regulates cognitive processes critical for goal-directed behavior. PFC cognitive dysfunction is implicated in multiple psychopathologies, including attention deficit hyperactivity disorder (ADHD). Although it has long been known that corticotropin-releasing factor (CRF) and CRF receptors are prominent in the PFC, the cognitive effects of CRF action within the PFC are poorly understood. The current studies examined whether CRF receptor activation in the PFC modulates cognitive function in rats as measured in a delayed response task of spatial working memory. CRF dose-dependently impaired working memory performance when administered either intracerebroventricularly (ICV) or directly into the PFC. The working memory actions of CRF in the PFC were topographically organized, with impairment observed only following CRF infusions into the caudal dorsomedial PFC (dmPFC). Additional studies examined whether endogenous CRF modulates working memory. Both ICV and intra-dmPFC administration of the nonselective CRF antagonist, D-Phe-CRF, dose-dependently improved working memory performance. To better assess the translational potential of CRF antagonists, we examined the cognitive effects of systemic administration of the CRF1 receptor selective antagonist, NBI 35965. Similar procognitive actions were observed in these studies. These results are the first to demonstrate that CRF acts in the PFC to regulate PFC-dependent cognition. Importantly, the ability of CRF antagonists to improve working memory is identical to that seen with all approved treatments for ADHD. These observations suggest that CRF antagonists may represent a novel approach for the treatment of ADHD and other disorders associated with dysregulated prefrontal cognitive function.


Women's satisfaction with physicians' communication skills during an infertility consultation.
The purpose of this study was to assess women's satisfaction with the communication of medical residents, in a university tertiary service in Brazil. The follow-up visits of 124 women consulting with 7 physicians were observed. An exit interview was performed to assess women's perception of their interaction with physicians. Factor analysis was used to build a satisfaction score, which was then studied according to the presence or absence (yes/no) of physicians' communicative behaviors observed by the researcher during consultation. Median satisfaction score was 9.4 in a 0-10 scale. Four physicians' communicative behaviors showed significant differences in the general satisfaction score: introducing oneself, setting the main reason for the visit, providing information about treatment procedures and asking whether patient had other issues or concerns. No significant differences were found in satisfaction scores for women's socialdemographic characteristics, reproductive history or duration of consultation. These results are important in the development of educational strategies for doctors and patients.


Optimal foraging: A case for random movement.
The bumblebee, Bombus flavifrons, forages randomly with respect to direction on Polemonium foliosissimum. This foraging pattern is as predicted for a system where there is a low probability of revisiting any given flower upon returning to a patch. This low revisitation probability is a function of the floral resource arrangement. It is further shown that B. flavifrons is using the resource distribution to direct its movements. A large percentage of all movements are to nearest neighbors with maximal foraging efficiency gained through minimization of flight distances.


Combined effects of prenatal phthalate exposure on cardiometabolic risk score among 4- to 7-year-old children: MABC study.
There is currently no consensus about the impact of prenatal phthalate exposure on blood pressure and glycolipids in children. Few studies consider the health effects as an integrated indicator. The combined effect of multiple phthalate exposures is often ignored. Based on the Ma'anshan Birth Cohort, 2298 woman-child pairs were included in this study. Maternal urine was collected in each trimester to analyze 6 phthalate metabolites. The overall cardiometabolic risk (CMR) score was calculated based on serum glycolipids and blood pressure for children aged 4-7 years. A higher score represents a less favorable CMR profile. The restricted cubic spline model was used to explore the relationship between prenatal phthalate exposure and childhood CMR score. In addition, the quantile g-computation and the Bayesian kernel machine regression were used to evaluate the combined effect. The MBP exposure in the 1st trimester (MBP-1st) and the MEP-2nd were non-linearly associated with the CMR score (Fnonlinear = 3.28 and 5.60, Pnonlinear = 0.0378 and 0.0038, respectively). The MBP-3rd (Flinear = 5.31, Plinear = 0.0012) and the ∑LMWP-3rd (Flinear = 4.37, Plinear = 0.0045) were negatively associated with the score in a linear manner. The phthalate mixture in the 2nd trimester increased the score (psil = 0.1747, 95% CI = 0.0077-0.3416), with the MEP being the most common [weights = 0.5290; posterior inclusion probability (PIP) = 0.40]. The phthalate mixture in the 3rd trimester decreased the score (psil = -0.2024, 95% CI = -0.4097-0.0048), with the MEHP (weights = -0.5101; PIP = 0.14) and the MBP (weights = -0.3993, PIP = 1.00) being the greatest contributors. In conclusion, the MBP-1st and the MEP-2nd are non-linearly associated with the cardiometabolic risk in children. The MBP-3rd and the ∑LMWP-3rd decrease the childhood risk. The combined exposure to phthalate mixture in the second and third trimester elevates and decreases the risk of childhood cardiometabolism, respectively.


Study of nociceptive flexion reflex in healthy subjects and patients with chronic neuropathic pain syndrome.
Nociceptive flexion reflex was measured in healthy subjects and patients with chronic neuropathic pain (diabetic distal symmetric sensorimotor polyneuropathy). The study of nociceptive flexion reflex revealed reduction of subjective pain threshold and reflex threshold in patients compared with healthy persons reflecting deficit of descending antinociceptive influences in the CNS.


DIPG-like MYB-altered diffuse astrocytoma with durable response to intensive chemotherapy.
Pontine gliomas represent difficult to treat entity due to the location and heterogeneous biology varying from indolent low-grade gliomas to aggressive diffuse intrinsic pontine glioma (DIPG). Making the correct tumor diagnosis in the pontine location is thus critical. Here, we report a case study of a 14-month-old patient initially diagnosed as histone H3 wild-type DIPG. Due to the low age of the patient, the MRI appearance of DIPG, and anaplastic astrocytoma histology, intensive chemotherapy based on the HIT-SKK protocol with vinblastine maintenance chemotherapy was administered. Rapid clinical improvement and radiological regression of the tumor were observed with nearly complete remission with durable effect and excellent clinical condition more than 6.5 years after diagnosis. Based on this unexpected therapeutic outcome, genome-wide DNA methylation array was employed and the sample was classified into the methylation class "Low-grade glioma, MYB(L1) altered." Additionally, RT-PCR revealed the presence of MYB::QKI fusion. Taken together, the histopathological classification, molecular-genetic and epigenetic features, clinical behavior, and pontine location have led us to reclassify the tumor as a pontine MYB-altered glioma. Our case demonstrates that more intensive chemotherapy can achieve long-term clinical effect in the treatment of MYB-altered pontine gliomas compared to previously used LGG-based regimens or radiotherapy. It also emphasizes the importance of a biopsy and a thorough molecular investigation of pontine lesions.


Real-space observation of ultraconfined in-plane anisotropic acoustic terahertz plasmon polaritons.
Thin layers of in-plane anisotropic materials can support ultraconfined polaritons, whose wavelengths depend on the propagation direction. Such polaritons hold potential for the exploration of fundamental material properties and the development of novel nanophotonic devices. However, the real-space observation of ultraconfined in-plane anisotropic plasmon polaritons (PPs)-which exist in much broader spectral ranges than phonon polaritons-has been elusive. Here we apply terahertz nanoscopy to image in-plane anisotropic low-energy PPs in monoclinic Ag2Te platelets. The hybridization of the PPs with their mirror image-by placing the platelets above a Au layer-increases the direction-dependent relative polariton propagation length and the directional polariton confinement. This allows for verifying a linear dispersion and elliptical isofrequency contour in momentum space, revealing in-plane anisotropic acoustic terahertz PPs. Our work shows high-symmetry (elliptical) polaritons on low-symmetry (monoclinic) crystals and demonstrates the use of terahertz PPs for local measurements of anisotropic charge carrier masses and damping.


Safeguard pressure assisted device for local femoral hemostasis in neuroendovascular procedures: A single center study of 879 patients.
Various vascular closure devices (VCDs) are available for local hemostasis after percutaneous transfemoral approach for neuroendovascular procedures but they have been associated with an increased complication rate and limitations to a re-puncture of the artery. We evaluated the safety and efficacy of Safeguard® 24 cm pressure assisted device (Merit Medical, West Jordan, UT, USA) and the associated complications. From September 2016 to December 2019, 879 patients underwent neuroendovascular procedures via transfemoral approach using an introducer sheath ranging from 4 to 6-French and they were included in a prospective database. We registered the demographic characteristics and procedural factors. We evaluated the device failure and associated complications. The Safeguard® was successful in 862 cases (98.1 %) with post-procedural local bleeding in 17 patients (1.9%). On univariate analysis, an association with local bleeding was observed with age >60 years (Odds ratio [OR] = 3.2, P = 0.04) and the use of an introducer sheath >4 F ([OR] = 3.1, P = 0.007). Female gender, antithrombotic medication and type of procedure (diagnostic or interventional) were not associated with local bleeding. On binary logistic regression analysis, there was association only for age >60 years ([OR] = 3, P = 0.04). The Safeguard® 24 cm is safe and efficient. It is simple to use and it can be applied independently from vessel anatomic characteristics. It should though be used with caution in case of a femoral introducer sheath larger than 4 Fr and patients older than 60 years.


Regulating Macrophage Polarization in High Glucose Microenvironment Using Lithium-Modified Bioglass-Hydrogel for Diabetic Bone Regeneration.
Diabetes mellitus is a chronic metabolic disease with a proinflammatory microenvironment, causing poor vascularization and bone regeneration. Due to the lack of effective therapy and one-sided focus on the direct angiogenic properties of biomaterials and osteogenesis stimulation, the treatment of diabetic bone defect remains challenging and complex. In this study, using gelatin methacryloyl (GelMA) as a template, a lithium (Li) -modified bioglass-hydrogel for diabetic bone regeneration is developed. It exhibits a sustained ion release for better bone regeneration under diabetic microenvironment. The hydrogel is shown to be mechanically adaptable to the complex shape of the defect. In vitro, Li-modified bioglass-hydrogel promoted cell proliferation, direct osteogenesis, and regulated macrophages in high glucose (HG) microenvironment, with the secretion of bone morphogenetic protein-2 and vascular endothelial growth factor to stimulate osteogenesis and neovascularization indirectly. In vivo, composite hydrogels containing GelMA and Li-MBG (GM/M-Li) release Li ions to relieve inflammation, providing an anti-inflammatory microenvironment for osteogenesis and angiogenesis. Applying Li-modified bioglass-hydrogel, significantly enhances bone regeneration in a diabetic rat bone defect. Together, both remarkable in vitro and in vivo outcomes in this study present an opportunity for diabetic bone regeneration on the basis of HG microenvironment.


Granule-Dependent NK Cell Killing of Cryptococcus Requires Kinesin to Reposition the Cytolytic Machinery for Directed Cytotoxicity.
Cryptococcus is the most important cause of fungal meningitis in immunocompromised individuals. Host defense against Cryptococcus involves direct killing by NK cells. That NK cells from HIV-infected patients fail to polarize perforin to the microbial synapse and kill C. neoformans led us to explore the mechanisms used to reposition and polarize the cytolytic granules to the synapse. Using live-cell imaging, we observed microtubule and granule movements in response to Cryptococcus that revealed a kinesin-dependent event. Eg5-kinesin bound to perforin-containing granules and was required for association with the microtubules. Inhibition of Eg5-kinesin abrogated dynein-dependent granule convergence to the MTOC and granule and MTOC polarization to the synapse and suppressed NK cell killing of Cryptococcus. In contrast, Eg5-kinesin was dispensable for tumor killing. This reveals an alternative mechanism of MTOC repositioning and granule polarization, not used in tumor cytotoxicity, in which Eg5-kinesin is required to initiate granule movement, leading to microbial killing.


Enhancing anoxic denitrification of low C/N ratio wastewater with novel ZVI composite carriers.
External organic carbon sources are needed to provide electron donors for the denitrification of wastewater with a low COD/NO3--N (C/N) ratio, increasing the treatment cost. The economic strategy is to enhance the bioactivity and/or biodiversity of denitrifiers to efficiently utilize organic substances in wastewater. In this study, novel zero-valent iron (ZVI) composite carriers were prepared and implemented in a suspended carrier biofilm reactor to enhance the bioactivity and/or biodiversity of denitrifiers. At the influent C/N ratio of 4 (COD was 179.5 ± 5.0 mg/L and TN was 44.2 ± 0.8 mg/L), COD and TN removal efficiencies were 85.1% and 66.4%, respectively, in the reactors filled with 3 wt% ZVI composite carriers. In contrast, COD and TN removal efficiencies were 70.4% and 55.3%, respectively, in the reactor filled with conventional high-density polyethylene (HDPE) biofilm carriers. The biofilm formation on the 3 wt% ZVI composite carriers was optimized due to its higher roughness (surface square roughness increased from 76.0 nm to 93.8 nm) and favorable hydrophilicity (water contact angle dropped to 72.5° ± 1.4° from 94.3° ± 3.2°) compared with the HDPE biofilm carriers. In addition, heterotrophic denitrifiers, Thauera and Dechloromonas, were enriched, whereas autotrophic denitrifiers, Raoultella and Thiobacillus, exhibited high relative abundance in the biofilm of ZVI composite carriers. The coexistence of heterotrophic denitrifiers and autotrophic denitrifiers on the surface of ZVI composite carriers provided mixotrophic metabolism of denitrification (including heterotrophic and iron-based autotrophic), thereby ensuring effective denitrification for wastewater with a low C/N ratio without external organic carbon source addition.


Effect of calcium formate as an accelerator on dilatancy deformation, strength and microstructure of cemented tailings backfill.
Recycling mining wastes to produce cemented tailings backfill (CTB) is the optimal approach to eliminate the environmental pollution caused by their accumulation. However, its low strength limits its application. Using calcium formate (CF) as an accelerator for improving its mechanical properties is of great significance to promote sustainable development. The effects of CF dosage and curing time on dilatancy deformation, compressive strength and microstructure of CTB were investigated through mechanical compression, scanning electron microscope (SEM) and energy dispersive spectrometer (EDS) tests. The strengthening and deterioration mechanisms of CF dosage on CTB were revealed, and its engineering practicability was systematically evaluated. The results show that the variation of volumetric strain in the dilatancy deformation stage firstly increase and then decrease with the increases of CF dosage and curing time. The relationship between CF dosage and compressive strength can be characterized by quadratic polynomial, and the optimal CF dosage characterizing the superior mechanical property of CTB is between 1.60 and 1.84. The supplement of CF reduces the size and distribution of microcracks and micropores, thereby optimizing the microstructure of CTB. Nevertheless, the excessive dosages of CF deteriorate the microstructure of CTB and produce serious defects, which cannot be effectively filled by hydration products, thus weakening the strength property of CTB. This study provides an effective accelerator for improving the mechanical properties of CTB, which is of great significance to promote the recycling of tailings.


The effect of inulin-type fructans on the intestinal immune function of antibiotic-treated mice.
This study aimed to evaluate the effect of supplementation with inulin-type fructans (ITFs) on the intestinal immune function in the context of dysbiosis resulting from antibiotic cocktail (ABx) treatment. BALB/c mice (8-9 weeks of age) were treated with an ABx for 3 weeks and then allowed to recover spontaneously or with ITF supplementation (5%) for 4 weeks. Our results showed that ABx treatment can induce gut microbiota dysbiosis and intestinal inflammation in mice. After 4 weeks of recovery, ITF supplementation restored the composition of the intestinal microbial community. However, compared with spontaneous recovery, ITF supplementation delayed inflammation recovery in the intestine and upregulated diamine oxidase (DAO) activity and increased lipopolysaccharide (LPS) content in serum. In addition, ITF supplementation delayed the regulatory T (Treg) cell and B cell recovery in the lamina propria (LP). Furthermore, compared with spontaneous recovery, ITF supplementation inhibited the relative expression of certain proinflammatory genes, such as for inducible nitric oxide synthase (iNOS) and tumour necrosis factor α (Tnf-α), in the colon, but it reduced the secretion of the anti-inflammatory mediator transforming growth factor β1 (TGF-β1) in serum, reduced the secretion of secretory immunoglobulin A (SIgA) in the colon and promoted the secretion of the proinflammatory cytokine interleukin (IL)-17A. In conclusion, these data supported the hypothesis that the influence of ITFs on the host's intestinal status is not always beneficial in the context of ABx-induced biological disorder. However, the significance of these findings needs to be determined by advanced studies KEY POINTS: • ITFs did not promote the recovery of microbial community composition. • ITFs delayed the recovery of ABx-induced colonic inflammation. • ITFs reduced the secretion of TGF-β1 and SIgA. • ITFs delayed the recovery of Treg and B cells in the LP.


Involvement of endogenous enkephalins in the feeding response to diazepam.
The effect of the delta-opioid agonist [D-Ala2,D-Leu5]enkephalin (DADLE) and the benzodiazepine diazepam on the food intake of non-deprived rats was investigated. Both compounds produced a significant increase in feeding. The hyperphagic response to DADLE (10 micrograms i.c.v.) and diazepam (1 mg/kg i.p.) was suppressed by concurrent treatment with the selective delta-opioid antagonist ICI 154,129 (10 micrograms i.c.v.). These findings add further support to the concept that endogenous enkephalins may be implicated in the stimulatory effects of benzodiazepines on ingestive behaviour.


Anti-leishmania effector functions of CD4+ Th1 cells and early events instructing Th2 cell development and susceptibility to Leishmania major in BALB/c mice.
Resistance and susceptibility to infection with the intracellular parasite, Leishmania major, are mediated by parasite-specific CD4+ Th1 and Th2 cells, respectively. It is well established that the protective effect of parasite-specific CD4+ Th1 cells is largely dependent upon the IFN-gamma produced. However, recent results indicate that the effect of Th1 cells on resolution of lesions induced by L. major in genetically resistant mice also requires a functional Fas-FasL pathway of cytotoxicity. In contrast to resistant mice, susceptible BALB/c mice develop aberrant Th2 responses following infection with L. major and consequently suffer progressive disease. These outcomes clearly depends upon the production of interleukin 4 (IL-4) early after infection. We have shown that a burst of IL-4 mRNA, peaking in draining lymph nodes of BALB/c mice 16 hrs after infection, occurs within CD4+ T cells that express V beta 4-V alpha 8 T cell receptors. In contrast to control and V beta 6-deficient mice, V beta 4-deficient BALB/c mice were resistant to infection, demonstrating the role of these cells in Th2 development. The early IL-4 response was absent in these mice, and Th1 responses occurred following infection. The LACK antigen of L. major induced comparable IL-4 production in V beta 4-V alpha 8 CD4+ T cells. Thus, the IL-4 required for Th2 development and susceptibility to L. major is produced by a restricted population of V beta 4-V alpha 8 CD4+ T cells after cognate interaction with a single antigen from this complex parasite. The IL-4 produced rapidly by these CD4+ T cells induces within 48 hours a state of unresponsiveness to IL-12 among parasite-specific CD4+ T cell precursors by downregulating the IL-12 receptor beta 2 chain expression.


An ethyl methanesulfonate-induced neutral mutant-bridging method efficiently identifies spontaneously mutated genes in rice.
Spontaneous mutants are mainly obtained from tissue culture or natural occurrences in plants. The traditional strategy for identifying spontaneously mutated genes is to continuously backcross these mutants to another variety and develop a near-isogenic F2 population for map-based cloning or bulked segregant analysis. However, this strategy is time-consuming. Here, we have developed a new method to efficiently accelerate the identification process. The chemical mutagen ethyl methanesulfonate was first used to treat the wild type of the spontaneous mutants to induce thousands of neutral mutations. An induced individual without any statistically significant phenotypic changes which was compared with the wild type was chosen as the neutral mutant. The spontaneous mutant was then crossed with the neutral mutant to develop a pseudo-near-isogenic F2 population in which only the induced neutral mutations and the causal mutation were segregated in the genome. This population ensures that the variation of the mutated trait is controlled only by the spontaneously mutated gene. Finally, after sequencing the neutral mutant and the mutant-type DNA pool of the F2 population the spontaneous mutation will be identified quickly by bioinformatics analysis. Using this method, two spontaneously mutated genes were identified successfully. Therefore, the neutral mutant-bridging method efficiently identifies spontaneously mutated genes in rice, and its value in other plants is discussed.