Bacterial cGAS-like enzymes produce 2',3'-cGAMP to activate an ion channel that restricts phage replication.
The mammalian innate immune system uses cyclic GMP-AMP synthase (cGAS) to synthesize the cyclic dinucleotide 2',3'-cGAMP during antiviral and antitumor immune responses. 2',3'-cGAMP is a nucleotide second messenger that initiates inflammatory signaling by binding to and activating the stimulator of interferon genes (STING) receptor. Bacteria also encode cGAS/DncV-like nucleotidyltransferases (CD-NTases) that produce nucleotide second messengers to initiate antiviral (antiphage) signaling. Bacterial CD-NTases produce a wide range of cyclic oligonucleotides but have not been documented to produce 2',3'-cGAMP. Here we discovered bacterial CD-NTases that produce 2',3'-cGAMP to restrict phage replication. Bacterial 2',3'-cGAMP binds to CD-NTase associated protein 14 (Cap14), a transmembrane protein of unknown function. Using electrophysiology, we show that Cap14 is a chloride-selective ion channel that is activated by 2',3'-cGAMP binding. Cap14 adopts a modular architecture, with an N-terminal transmembrane domain and a C-terminal nucleotide-binding SAVED domain. Domain-swapping experiments demonstrated the Cap14 transmembrane region could be substituted with a nuclease, thereby generating a biosensor that is selective for 2',3'-cGAMP. This study reveals that 2',3'-cGAMP signaling extends beyond metazoa to bacteria. Further, our findings suggest that transmembrane proteins of unknown function in bacterial immune pathways may broadly function as nucleotide-gated ion channels.


A nanotherapeutic strategy to overcome chemoresistance to irinotecan/7-ethyl-10-hydroxy-camptothecin in colorectal cancer.
Clinical development of 7-ethyl-10‑hydroxy-camptothecin (SN38), the active metabolite of irinotecan (CPT-11), is hindered by its insolubility and poor stability. Another obstacle is that tumors could become resistant to SN38/CPT-11 through multiple mechanisms involving breast cancer resistance protein (BCRP). Herein one of the most potent and selective BCRP inhibitors, Ko143, is encapsulated into a recently constructed prodrug PEG-S-S-SN38 displaying a high and fixed drug loading, multiple intratumoral stimuli (oxidative stress, GSH and esterase)-responsive drug release and significant in vitro and in vivo superiorities over CPT-11. The obtained "combo" for simultaneous delivery of SN38 and Ko143, named as BI@PEG-SN38, has a high SN38 loading efficacy (14.85 wt.%) and a good Ko143 encapsulation efficacy (3.79%). Through generating panels of human colorectal cancer models expressing altered levels of BCRP via lentiviral transfection and CRISPR-Cas9, characteristics of different drug formulations are carefully evaluated. Impressively, BI@PEG-SN38 nanoparticles effectively reverse chemoresistance to CPT-11 (resistance index dropping from ∼274.00-456.00 to ∼1.70-4.68) and PEG-S-S-SN38 (resistance index dropping from ∼5.83-14.00 to ∼1.70-4.68) in three BCRP-overexpressing cancer cell lines. More importantly, reversal of BCRP-mediated chemoresistance to CPT-11 (P values lower than 0.001-0.0001) and PEG-S-S-SN38 (P values lower than 0.01-0.001) by BI@PEG-SN38 nanoparticles are further confirmed with two panels of colorectal cancer xenograft models in vivo. As the first nano-formulation of Ko143 and the first systemic co-delivery vehicle of SN38/CPT-11 and a BCRP inhibitor, BI@PEG-SN38 provides a new approach for resolving the bottlenecks for clinical translation of SN38 and numerous "chemosensitizers" like Ko143, and exhibits promising applicability in precision cancer medicine. STATEMENT OF SIGNIFICANCE: To resolve the bottlenecks in clinical application of anticancer agents SN38/CPT-11 and the most potent breast cancer resistant protein (BCRP) inhibitor Ko143, a "combo" nanotherapeutic simultaneously delivering SN38 and Ko143 was constructed and named as BI@PEG-SN38. By generating panels of colorectal cancer models, we demonstrate that BI@PEG-SN38 nanoparticles effectively and selectively reversed BCRP-mediated tumor resistance to SN38/CPT-11 in vitro and in vivo. As the first nano-formulation of Ko143 and the first systemic co-delivery vehicle of SN38/CPT-11 and a BCRP inhibitor, BI@PEG-SN38 provides a new strategy for clinical development of SN38 and numerous "chemosensitizers", and exhibits promising applicability in precision cancer medicine. Panels of cancer cell lines established here provides a useful platform for BCRP- and cancer-related research and technology development.


Immediate Shockwave Lithotripsy vs Delayed Shockwave Lithotripsy After Urgent Ureteral Stenting in Patients with Ureteral or Pyeloureteral Urolithiasis: A Matched-Pair Analysis.
Introduction: The most common cause of acute renal colic is a ureteral obstruction caused by ureterolithiasis. Urgent intervention is often necessary due to intractable pain. Early extracorporeal shockwave lithotripsy (SWL) as an alternative treatment option to ureteral stenting becomes forgotten in times of rising ureterorenoscopy. However, definitive guidelines are lacking, in which urgent treatment should be preferred in the absence of signs of infection. Therefore, we assessed efficacy and safety of early SWL (eSWL) to secondary SWL (sSWL) after urgent ureteral stenting. Patients and Methods: One hundred four patients treated between January 2015 and November 2017 for obstructive ureterolithiasis were matched regarding stone size, stone localization, and assigned to group eSWL (n = 52) or group sSWL (n = 52). The eSWL group received shock waves (without prior ureteral stenting) and sSWL group ureteral stenting within 48 hours from diagnosis. Thereafter, patients in group sSWL were treated with shock waves for a median of 23 ± 14.6 days after ureteral stenting. Stone-free rates, complication rates, and reintervention rates were assessed. Univariable and multivariable logistic regression was applied to find predictors of outcomes in the two treatment groups. Results: Overall, there was no statistically significant difference between both groups regarding stone-free rate and complication rate. Reinterventions were more often addressed for patients in group sSWL (p = 0.05). eSWL was significantly superior to sSWL regarding stone-free rates for stones between 6 and 9 mm (p = 0.04). At the multivariable multinomial logistic regression none of the two treatment modalities was associated with better outcomes. A body mass index ≥30 was associated with a reduced 6-week stone-free status (p = 0.04), whereas stones ≥8 mm were associated with an increased need of reintervention (p = 0.04). Conclusion: eSWL seems to be an effective and safe emergency procedure compared with sSWL after urgent stenting within 6 weeks and should be considered as a treatment option in patients without absolute indications for immediate ureteral drainage. Clinical trial registration number: 2019-00155.


Quality of life after breast cancer surgery.
All patients (N = 41) who participated in a randomized clinical trial in the Leiden University Hospital comparing mastectomy to tumorectomy have been studied with respect to their quality of life 11 months (+/- 5) after surgery. The body image of patients is more severely impaired after mastectomy than after tumorectomy (P less than .01). The studied groups did not differ, as has been suggested, with respect to fear of recurrence and death. These fears, however, turned out to be inversely related to the age of the patients (r = -.34, P = .02).


Length and sequence heterogeneity of the histone gene repeat unit of the sea urchin, S. purpuratus.
Histone gene repeats in S. purpuratus are shown to be of variable length and sequence. Two recombinant plasmids containing the full-length 6.3 kb histone repeat unit are found to differer in length at two sites in the repeating structure and in the occurrence of two restriction enzyme recognition sites. Variation in repeat length is also demonstrated in the unfractionated DNA of five sea urchins and in a sample of DNA enriched for histone gene sequences by density gradient methods. The repeats in each individual are of a very limited number of major classes, which may differ from one another in overall length or in distribution and presence of particular restriction enzyme sites. Variations are found to occur at many regions of the repeat; some have been mapped specifically to spacer regions. Repeats may differ dramatically from individual to individual since there is no one type of repeat class common to all, although the absolute length differences of the repeats that are found are small.


On the Perils of Universal and Product-Led Thinking Comment on "How Neoliberalism Is Shaping the Supply of Unhealthy Commodities and What This Means for NCD Prevention".
Lencucha and Thow's paper offers an important addition and corrective to the burgeoning body of work in public health on the 'commercial determinants of health' in the context of non-communicable diseases (NCDs). Rather than tracing the origins of incoherence across policy sectors to the nefarious actions of industry, they argue that we need to be better attuned to the neoliberal ideologies that underpin these policies. In this commentary I explore two aspects of their argument that I find to be problematic: First, the suggestion that neoliberalism itself has some kind of deterministic or explanatory capacity across vastly different social, spatial, economic and political contexts. Second, I explore their concept of 'product-based NCD risk,' a perspective that disembodies and detaches risk from the social and structural conditions of their making.


A comparison between parent and grandparent dietary provision, feeding styles and feeding practices when caring for preschool-aged children.
Grandparents are frequently called upon to provide childcare to young children. Consequently, grandparents may influence the development of children's eating habits and preferences and may require support with their approach to feeding young children. However, research into grandparental feeding behaviours is scarce. Understanding how grandparental feeding behaviours compare to parental feeding behaviours will further help to establish whether grandparents require specific interventions unique to the grandparental role or if current strategies that target parental feeding behaviour are also appropriate for grandparents. The aim of the present study was to explore the similarities and differences between parent and grandparent dietary provision, feeding practices and feeding styles to preschool-aged children. 72 parents and 44 unrelated grandparents of children aged 2-4 years old took part in an online study and completed an online 24-h dietary recall using myfood24® to assess dietary provision. Parents and grandparents were providing meals high in saturated fat and sodium and providing below recommended amounts of fruit and vegetables. Overall, feeding practices were similar between parents and grandparents. Although, grandparents scored lower on using food as a reward (p < 0.05) and creating a healthy food environment (p < 0.05) compared to parents. Whereas, parents scored higher for promoting balance and variety (p < 0.05). A range of feeding styles were found within each caregiver type, with no significant associations found between caregiver type and feeding style (p > 0.05). Strategies to promote healthy eating in young children should be expanded to also target grandparents who act as informal caregivers to preschool-aged children. However, since very few differences in feeding behaviour were reported the content of such strategies may not need to be adapted specifically for grandparents.


Low-molecular-weight heparin (reviparin) reduces the incidence of femoropopliteal in-stent stenosis: preliminary results of an ongoing study.
To examine the efficacy of the low-molecular-weight heparin, reviparin, for prevention of femoropopliteal stent restenosis. Forty-two patients who had implantation of flexible tantalum stents for the treatment of stenosis (n = 24) or occlusion (n = 18) of the femoral (n = 27) or popliteal (n = 15) arteries were included in this study protocol. An intraarterial bolus of 5000 IU heparin was given before percutaneous transluminal angioplasty (PTA), and in the case of stent implantation due to unsuccessful PTA, an additional dose of reviparin (3500 anti-factor Xa IU) was given. Postprocedurally, 10,500 anti-factor Xa IU of reviparin were administered intravenously over 24 hr, followed by 3500 anti-factor Xa IU subcutaneously twice a day for 23 days. Oral aspirin (100 mg/day) was prescribed for the long term. Follow-up criteria (maximum follow-up 37 months) were clinical symptoms, Doppler ankle arm indices, color and duplex sonography, and angiography for suspicion of restenosis. Early stent thromboses were not observed. Overall primary patency rate (PPR) was 88% +/- 6.0% (1 year) and 74% +/- 10.1% (2 years). Major hemorrhagic complications have not occurred. Reviparin administered in a high dose over a period of 24 days is a safe medication regimen and provides excellent patency rates after stent implantation.


Attainment of Targets of the 20-Year Infancy-Onset Dietary Intervention and Blood Pressure Across Childhood and Young Adulthood: The Special Turku Coronary Risk Factor Intervention Project (STRIP).
We examined whether success in achieving the key targets of an infancy-onset 20-year dietary intervention was associated with blood pressure (BP) from infancy to young adulthood. In the prospective randomized STRIP (Special Turku Coronary Risk Factor Intervention Project; n=877 children), dietary counseling was provided biannually based on the Nordic Nutrition Recommendations primarily to improve the quality of dietary fat in children's diets and secondarily to promote intake of vegetables, fruits, and whole grains. Dietary data and BP were accrued annually from the age of 13 months to 20 years. The dietary targets for fat quality were defined as the ratio of saturated fatty acids to monounsaturated and polyunsaturated fatty acids <1:2 and intake of saturated fatty acids <10 E%, dietary fiber intake in the top age-specific quintile, and dietary sucrose intake as being in the lowest age-specific quintile. Attaining a higher number of the dietary targets was associated with lower systolic BP (mean [SE] systolic BP, 107.3 [0.3], 107.6 [0.3], 106.8 [0.3], and 106.7 [0.5] mm Hg in participants meeting 0, 1, 2, and 3 to 4 targets, respectively; P=0.03) and diastolic BP (mean [SE] diastolic BP, 60.4 [0.2], 60.5 [0.2], 59.9 [0.2], and 59.9 [0.3] mm Hg; P=0.02). When the lowest age-specific quintile of dietary cholesterol was added as an additional target, the association with systolic BP remained significant (P=0.047), but the association with diastolic BP attenuated (P=0.13). Achieving the key targets of an infancy-onset 20-year dietary intervention, reflecting dietary guidelines, was favorably albeit modestly associated with systolic and diastolic BP from infancy to young adulthood. Registration- URL: https://www.clinicaltrials.gov; Unique identifier: NCT00223600.


Novel joint selection methods can reduce sample size for rheumatoid arthritis clinical trials with ultrasound endpoints.
To determine whether novel methods of selecting joints through (i) ultrasonography (individualized-ultrasound [IUS] method), or (ii) ultrasonography and clinical examination (individualized-composite-ultrasound [ICUS] method) translate into smaller rheumatoid arthritis (RA) clinical trial sample sizes when compared to existing methods utilizing predetermined joint sites for ultrasonography. Cohen's effect size (ES) was estimated (ES^) and a 95% CI (ES^L, ES^U) calculated on a mean change in 3-month total inflammatory score for each method. Corresponding 95% CIs [nL(ES^U), nU(ES^L)] were obtained on a post hoc sample size reflecting the uncertainty in ES^. Sample size calculations were based on a one-sample t-test as the patient numbers needed to provide 80% power at α = 0.05 to reject a null hypothesis H0 : ES = 0 versus alternative hypotheses H1 : ES = ES^, ES = ES^L and ES = ES^U. We aimed to provide point and interval estimates on projected sample sizes for future studies reflecting the uncertainty in our study ES^S. Twenty-four treated RA patients were followed up for 3 months. Utilizing the 12-joint approach and existing methods, the post hoc sample size (95% CI) was 22 (10-245). Corresponding sample sizes using ICUS and IUS were 11 (7-40) and 11 (6-38), respectively. Utilizing a seven-joint approach, the corresponding sample sizes using ICUS and IUS methods were nine (6-24) and 11 (6-35), respectively. Our pilot study suggests that sample size for RA clinical trials with ultrasound endpoints may be reduced using the novel methods, providing justification for larger studies to confirm these observations.


Wnt signaling pathway in rheumatoid arthritis, with special emphasis on the different roles in synovial inflammation and bone remodeling.
Rheumatoid arthritis (RA) is a chronic symmetrical autoimmune disease of unknown etiology that affects primarily the diarthrodial joints. Characteristic features of RA pathogenesis are synovial inflammation and proliferation accompanied by cartilage erosion and bone loss. Fibroblast-like synoviocytes (FLS) display an important role in the pathogenesis of RA. Several lines of evidence show that the Wnt signaling pathway significantly participates in the RA pathogenesis. The Wnt proteins are glycoproteins that bind to the Fz receptors on the cell surface, which leads to several important biological functions, such as cell differentiation, embryonic development, limb development and joint formation. Accumulated evidence has suggested that this signaling pathway plays a key role in the FLS activation, bone resorption and joint destruction during RA development. Greater knowledge of the role of the Wnt signaling pathway in RA could improve understanding of the RA pathogenesis and the differences in RA clinical presentation and prognosis. In this review, new advances of the Wnt signaling pathway in RA pathogenesis are discussed, with special emphasis on its different roles in synovial inflammation and bone remodeling. Further studies are needed to reveal the important role of the members of the Wnt signaling pathway in the RA pathogenesis and treatment.


Predicting Functional Interactions Among Genes in Prokaryotes by Genomic Context.
Genomic context methods for finding functions of unannotated genes were implemented very early after the publication of the first few prokaryotic genomes. The ideas behind these methods include gene fusions, conservation of gene adjacency, and the patters of co-occurrence of genes across available genomes. A later addition was the prediction of features related to functional organization, such as operons, stretches of genes co-transcribed into a single messenger RNA. The ideas behind these methods tend to be easy to understand, while the strategies for transforming those basic ideas into predictions can vary in complexity, mostly because genes whose products are known to functionally interact vary in the way they relate to those basic ideas. We present here a view of genomic context methods for predicting functional interactions, with simple examples of their implementation as compared and evaluated using genes whose products are known to functionally interact.


Respiratory mechanics by least squares fitting in mechanically ventilated patients: applications during paralysis and during pressure support ventilation.
To evaluate a least squares fitting technique for the purpose of measuring total respiratory compliance (Crs) and resistance (Rrs) in patients submitted to partial ventilatory support, without the need for esophageal pressure measurement. Prospective, randomized study. A general ICU of a University Hospital. 11 patients in acute respiratory failure, intubated and assisted by pressure support ventilation (PSV). Patients were ventilated at 4 different levels of pressure support. At the end of the study, they were paralyzed for diagnostic reasons and submitted to volume controlled ventilation (CMV). A least squares fitting (LSF) method was applied to measure Crs and Rrs at different levels of pressure support as well as in CMV. Crs and Rrs calculated by the LSF method were compared to reference values which were obtained in PSV by measurement of esophageal pressure, and in CMV by the application of the constant flow, end-inspiratory occlusion method. Inspiratory activity was measured by P0.1. In CMV, Crs and Rrs measured by the LSF method are close to quasistatic compliance (-1.5 +/- 1.5 ml/cmH2O) and to the mean value of minimum and maximum end-inspiratory resistance (+0.9 +/- 2.5 cmH2O/(l/s)). Applied during PSV, the LSF method leads to gross underestimation of Rrs (-10.4 +/- 2.3 cmH2O/(l/s)) and overestimation of Crs (+35.2 +/- 33 ml/cmH2O) whenever the set pressure support level is low and the activity of the respiratory muscles is high (P0.1 was 4.6 +/- 3.1 cmH2O). However, satisfactory estimations of Crs and Rrs by the LSF method were obtained at increased pressure support levels, resulting in a mean error of -0.4 +/- 6 ml/cmH2O and -2.8 +/- 1.5 cmH2O/(l/s), respectively. This condition was coincident with a P0.1 of 1.6 +/- 0.7 cmH2O. The LSF method allows non-invasive evaluation of respiratory mechanics during PSV, provided that a near-relaxation condition is obtained by means of an adequately increased pressure support level. The measurement of P0.1 may be helpful for titrating the pressure support in order to obtain the condition of near-relaxation.


Electrical conduction in some nucleic acid base complexes with acridine orange dye: study of localized levels.
Semiconduction activation energies of adenine, guanine, thymine and uracil complexes with acridine orange dye at various dye concentrations were measured in both Ohmic and space charge limited (SCL) current regimes. The behaviour of the complexes is non-extrinsic at high dye concentrations but extrinsic at low dye concentrations. Dominant electron and hole levels are shown to depend also on dye concentration. Various transport parameters have been evaluated.


The Neurological Evaluation Scale (NES): a structured instrument for the assessment of neurological signs in schizophrenia.
The significance of neurological signs in schizophrenia is poorly understood. In part, this may reflect the marked variability in the methods of ascertainment in previous studies. The Neurological Evaluation Scale (NES) is designed to standardize the assessment of neurological impairment in schizophrenia. The battery consists of 26 items. Data on the interrater reliability for total score, functional areas of interest, and individual items are presented. Preliminary validity data demonstrate the ability of the battery to discriminate patients with schizophrenia from nonpsychiatric controls.


Serological survey for Aujeszky's disease in native sows of Canada.
A serological survey was initiated to uncover subclinical foci of infection with pseudorabies virus. A total of 2819 serum samples, collected from sows during February to November 1977, were found negative for antibodies to pseudorabies virus.


Differential activation of p38 and extracellular signal-regulated kinase in spinal cord in a model of bee venom-induced inflammation and hyperalgesia.
Honeybee's sting on human skin can induce ongoing pain, hyperalgesia and inflammation. Injection of bee venom (BV) into the intraplantar surface of the rat hindpaw induces an early onset of spontaneous pain followed by a lasting thermal and mechanical hypersensitivity in the affected paw. The underlying mechanisms of BV-induced thermal and mechanical hypersensitivity are, however, poorly understood. In the present study, we investigated the role of mitogen-activated protein kinase (MAPK) in the generation of BV-induced pain hypersensitivity. We found that BV injection resulted in a quick activation of p38, predominantly in the L4/L5 spinal dorsal horn ipsilateral to the inflammation from 1 hr to 7 d post-injection. Phosphorylated p38 (p-p38) was expressed in both neurons and microglia, but not in astrocytes. Intrathecal administration of the p38 inhibitor, SB203580, prevented BV-induced thermal hypersensitivity from 1 hr to 3 d, but had no effect on mechanical hypersensitivity. Activated ERK1/2 was observed exclusively in neurons in the L4/L5 dorsal horn from 2 min to 1 d, peaking at 2 min after BV injection. Intrathecal administration of the MEK inhibitor, U0126, prevented both mechanical and thermal hypersensitivity from 1 hr to 2 d. p-ERK1/2 and p-p38 were expressed in neurons in distinct regions of the L4/L5 dorsal horn; p-ERK1/2 was mainly in lamina I, while p-p38 was mainly in lamina II of the dorsal horn. The results indicate that differential activation of p38 and ERK1/2 in the dorsal horn may contribute to the generation and development of BV-induced pain hypersensitivity by different mechanisms.


Comparison of the Big Five personality traits in fibromyalgia and other rheumatic diseases.
The personality of patients with fibromyalgia is still under debate. Some studies found high neuroticism associated with low extraversion, while others found that these traits do not differ from the normal population. Personality factors intervene in the emotional regulation and modulation of pain. The aim of the study was to determine the personality traits of patients with fibromyalgia compared to other rheumatic diseases. In a multicentric study, women with fibromyalgia, rheumatoid arthritis, spondyloarthritis or Sjögren's syndrome were asked to complete the Big Five Inventory, which encompasses five main personality dimensions, namely (1) extraversion vs. introversion, (2) agreeableness vs. antagonism, (3) conscientiousness vs. impulsivity, (4) neuroticism vs. emotional stability, and (5) openness vs. closed-mindedness. Variance analysis (Student's t-test and ANOVA with post-hoc comparisons or Bonferroni correction) was performed. We also conducted hierarchical and non-hierarchical cluster analyses. Participants were 163 women with fibromyalgia (n=48), rheumatoid arthritis (n=46), spondyloarthritis (n=46) and Sjögren's syndrome (n=23). The mean age was 47.18years (±10.81years, range 21 to 65). Patients with fibromyalgia had higher scores on agreeableness (F(3, 159)=3.39, P<0.05), neuroticism (F(3, 159)=3.79, P<0.05) and openness (F(3, 159)=4.32, P<0.01) than those with other rheumatic diseases. This study highlights the specificity of personality in fibromyalgia. It also underlines the protective role of personality traits: in the fibromyalgia group, high neuroticism and low conscientiousness (high impulsivity) were associated with a high level of chronic pain.


Effects of ALA-PDT on the macrophages in wound healing and its related mechanisms in vivo and in vitro.
Several studies have suggested the effectiveness of photodynamic therapy (PDT) for wound healing. Macrophages are critical immune cells necessary for regulated inflammation during wound repair. However, the available information regarding the effects of PDT on macrophages during cutaneous wound healing remains insufficient. This study aimed to further investigate these aspects in vivo and in vitro. Mouse full-thickness wound models were used as the study samples to investigate the therapeutic effects and mechanisms of 5-aminolevulinic acid (ALA) PDT. Wound healing rate, granulation tissue formation, local inflammation, M1/M2 macrophages differentiation, were measured at different time points treated by ALA-PDT. The polarization of macrophages induced by ALA-PDT was further evaluated in vitro using PCR and western blot analysis. ALA-PDT could promote formation of granulation tissue, increase inflammatory infiltration and activate M1 macrophages in the early stage of injury. While, ALA-PDT could also facilitate absorption of granulation tissue, inhibit inflammatory infiltration and enhance M2 macrophages polarization in the later stage of wound repair. In vitro, ALA-PDT could modulate the ratio of M2 polarization to M1 polarization via NF-κB signaling pathway. ALA-PDT topical application stimulates wound healing by regulating formation of granulation tissue, inflammatory process and M1/M2 macrophages differentiation. The study places a preliminary theoretical basis for topical ALA-PDT to be administered clinically in cutaneous wounds healing.


An amplification technology for improving sensitivity when measuring components in biological samples.
A new technology for improving the sensitivity in measuring components in biological samples is described. The method is based on the use of spherical microbeads (detection beads) which contain a large amount of immobilized enzyme and a reagent with biospecific affinity for the component to be detected. These microbeads have been used in a 'sandwich reaction' for visualization of P-fimbriated Escherichia coli which has a known receptor structure (Gal(alpha 1-4)Gal(beta)). In the initial step the bacteria were enriched on a solid support (e.g., a plastic film or beads (greater than 150 microns)) to which the receptor structure had been covalently bound. In the next step detection beads coupled with enzyme and receptor structure were added and finally a chromogenic substrate for the enzyme was used for visualization. A sensitivity of 10(5) bacteria/ml was reached. The detection beads are of general utility and might be useful for the detection of lectins on other pathogens.


Origin of flaw-tolerance in nacre.
Over the past decades, our understanding of nacre's toughening origin has long stayed at the level of crack deflection along the biopolymer interface between aragonite platelets. It has been widely thought that the ceramic aragonite platelets in nacre invariably remain shielded from the propagating crack. Here we report an unexpected experimental observation that the propagating crack, surprisingly, invades the aragonite platelet following a zigzag crack propagation trajectory. The toughening origin of previously-thought brittle aragonite platelet is ascribed to its unique nanoparticle-architecture, which tunes crack propagation inside the aragonite platelet in an intergranular manner. For comparison, we also investigated the crack behavior in geologic aragonite mineral (pure monocrystal) and found that the crack propagates in a cleavage fashion, in sharp contrast with the intergranular cracking in the aragonite platelet of nacre. These two fundamentally different cracking mechanisms uncover a new toughening strategy in nacre's hierarchical flaw-tolerance design.


A randomized controlled clinical trial of a bereavement support group intervention in human immunodeficiency virus type 1-seropositive and -seronegative homosexual men.
Bereavement is a severe and frequent stressor among those infected with human immunodeficiency virus type 1 (HIV-1) and those affected by the acquired immunodeficiency syndrome epidemic. This study examined the impact of a research-derived, semistructured, bereavement support group among HIV-1-seropositive and HIV-1-seronegative homosexual men having lost a close friend or intimate partner to the acquired immunodeficiency syndrome within the prior 6 months. A total of 166 subjects (97 HIV-1 seropositive; 69 HIV- 1 seronegative) were randomly assigned to groups of homogeneous HIV-1 serostatus or to their respective control group. Subjects were assessed at entry and at 10 weeks with psychosocial questionnaires, a semistructured interview for psychopathology, a medical history and physical examination, urine collection, and phlebotomy. For a composite score of psychological distress and grief as well as the distress component, scores were significantly lower after the intervention by analyses against baseline scores, with and without control variables for other factors affecting distress level. A significant reduction in grief level was found only in the analysis that included control variables. Control subjects showed no significant decrements in overall distress, although a significant decrement in grief level was observed. A brief group intervention can significantly reduce overall distress and accelerate grief reduction in a sample of bereaved subjects unselected for psychopathology or at high risk for subsequent maladjustment.


Assessment of human placental perfusion by intravoxel incoherent motion MR imaging.
To provide functional information on the human placenta, including perfusion, and diffusion, with no contrast agent injection, and to study correlations between intravoxel incoherent motion (IVIM) placental parameters and fetal growth. MRI was performed in women undergoing legal termination of pregnancy at 17-34 weeks, including a 4-b-value and 11-b-value DW sequences. The apparent diffusion coefficient (ADC), the restricted diffusion coefficient (D), the pseudoperfusion coefficient (D*), and the perfusion fraction (f) were calculated. Their relationships with gestational age, Z-scores for fetal and placental weight were evaluated by means of regression analysis. Logistic regression analysis was used to assess the ability of IVIM parameters to predict/detect intrauterine growth retardation (SGA). Fifty-five pregnant women, including nine cases of SGA (16%), were included in the study. The ADC (n = 55) showed a quadratic correlation with gestational age (p < .001) and a linear correlation with the fetal weight Z-score (p = .02). Mean ADC values were significantly different between normally growing and SGA fetuses (2.37 ± 0.25 versus 2.29 ± 0.33 10-3.mm2.s-1, p=.048). The perfusion fraction f (n = 23) showed a quadratic correlation with gestational age (p = .017) and a linear correlation with the fetal weight Z - score (p = .008). Mean f values differed significantly between normally growing and SGA fetuses (42.55 ± 9.30% versus 27.94 ± 8.76%, p = .002). The receiver operating characteristics (ROC) curve for f to predict SGA was produced (area under the ROC curve = 0.9). The observed association between f and fetal weight suggests that fMRI could be suitable for studying placental insufficiency and for identifying risk of SGA.


16p11.2 haploinsufficiency reduces mitochondrial biogenesis in brain endothelial cells and alters brain metabolism in adult mice.
Neurovascular abnormalities in mouse models of 16p11.2 deletion autism syndrome are reminiscent of alterations reported in murine models of glucose transporter deficiency, including reduced brain angiogenesis and behavioral alterations. Yet, whether cerebrovascular alterations in 16p11.2df/+ mice affect brain metabolism is unknown. Here, we report that anesthetized 16p11.2df/+ mice display elevated brain glucose uptake, a phenomenon recapitulated in mice with endothelial-specific 16p11.2 haplodeficiency. Awake 16p11.2df/+ mice display attenuated relative fluctuations of extracellular brain glucose following systemic glucose administration. Targeted metabolomics on cerebral cortex extracts reveals enhanced metabolic responses to systemic glucose in 16p11.2df/+ mice that also display reduced mitochondria number in brain endothelial cells. This is not associated with changes in mitochondria fusion or fission proteins, but 16p11.2df/+ brain endothelial cells lack the splice variant NT-PGC-1α, suggesting defective mitochondrial biogenesis. We propose that altered brain metabolism in 16p11.2df/+ mice is compensatory to endothelial dysfunction, shedding light on previously unknown adaptative responses.


Molecularly imprinted polymer-based bulk optode for the determination of itopride hydrochloride in physiological fluids.
We report here for the first time on the use of Molecularly Imprinted Polymers as modifiers in bulk optodes, Miptode, for the determination of a pharmaceutical compound, itopride hydrochloride as an example in a concentration range of 1×10(-1)-1×10(-4)molL(-1). In comparison to the optode containing the ion exchanger only (Miptode 3), the optode containing the ion exchanger and the MIP particles (Miptode 2) showed improved selectivity over the most lipophilic species, Na(+) and K(+), by more than two orders of magnitude. For instance, the optical selectivity coefficients using Miptode 2, [Formula: see text] , were as follow: NH4(+)˂-6; Na(+)=-4.0, which were greatly enhanced in comparison with that obtained by Miptode 3. This work opens a new avenue for using miptodes for the determination of all the pharmaceutical preparations without the need for the development of new ionophores.


Management and prognostic factors of recurrent pleomorphic adenoma of the parotid gland: personal experience and review of the literature.
The aim of this study was to investigate the management and prognostic determinants of recurrent pleomorphic adenoma (RPA). A retrospective analysis was performed to examine the clinical features, the prevalence of surgical complications, and new recurrences of RPA. Tumor recurrence rate was estimated by the Kaplan-Meier method, and the prognostic value of some of the variables was tested by univariate analysis using the log rank test. The study focused on 33 patients, 18 female (54.5%) and 15 male (45.5%), aged 12-71 years (median 41). A total or extended total parotidectomy was performed in 16 cases (48.5%), a superficial parotidectomy in 10 cases (30.3%), and a local excision in 7 cases (21.2%). In ten patients (30.3%), a branch or the trunk of the facial nerve was deliberately sacrificed. Major complications included one unexpected definitive paralysis of the marginal mandibular branch of the facial nerve and 14 cases of Frey syndrome. Follow-up varied from 2 to 25 years (median 10.5 years), and there were 11 new recurrences (33.3%) within a period varying from 1 to 16 years (median 6 years). The estimated tumor recurrence rates were 14.1+/-6.6% at 5 years, 31.4+/-9.4% at 10 years, 43.0+/-10.8% at 15 years, and 57.2+/-14.8% at 20 years. Presence of a multinodular lesion and the type of intervention performed were significantly associated with a higher probability of recurrence. RPAs are prone to new recurrences, especially when multinodular and treated with a local excision. Surgical treatment should include facial nerve resection in selected cases. Follow-up for the patient's lifetime is warranted.


[Research progress of decellularized matrix in tissue engineered trachea transplantation].
To summarize the recent research situation and progress of decellularized matrix in tissue engineered trachea transplantation and to forecast the possible perspects. Recent original articles about study and application for decellularized matrix in tissue engineered trachea were reviewed. The application and study of different decellularized matrices involved in animals or patients with tracheal lesions were elaborated. Decellularized matrices researched and applied in tissue engineered trachea include jejunum, urinary bladder, aorta, and trachea. Decellularized urinary bladder matrix and jejunal matrix appears to be efficacious method for the patch repair of partial circumferential tracheal defects. The application of decellularized aortic matrix may need more study, and decellularized tracheal matrix has a bright future in long tracheal defects.


Novel strategy for f-HAp/PVP/Ag nanocomposite synthesis from fluoro based ionic liquid assistance: Systematic investigations on its antibacterial and cytotoxicity behaviors.
A novel biomimetic f-HAp/PVP/Ag nanocomposite was synthesized under the ionic liquid medium, which was composed of inorganic and organic nanofillers like fluor-hydroxyapatite, silver nanoparticles and polyvinyl pyrrolidone. In composite synthesis, the first time we were used fluorine based ionic liquid for the fluorine contents on the fluor-hydroxyapatite nanoparticles which were resulting in very good crosslinking and interfacial bonding with the PVP and Ag nanoparticles. Ionic liquid has assisted good morphological structure of both inorganic materials. The chemical interaction and crystallinity changes of the nanocomposite were evaluated by FTIR and XRD studies. The surface morphology and composition of the samples were observed by FE-SEM, HR-TEM and EDS analyses. This report reveals that the greener approach for synthesis of fluor-hydroxyapatite nanocomposite and sustained delivery of silver and fluorine ions from the fluor-hydroxyapatite surface to the bacterial surface have been reducing the bacterial growth rate which was evaluated by different pathogenic bacterial strains via different methods and it also favourable cytotoxicity effect with human osteosarcoma (MG-63) cells.


Preparation of 111In leukocytes after hemolytic removal of erythrocytes.
An optimized procedure is described for isolation and high-efficiency radiolabeling of leukocytes using 111In-oxine. The chief advantages over conventional methods include virtually no loss of leukocytes during washing and separation steps; a significant reduction in the time required to prepare leukocytes for radiolabeling compared to non-hemolytic preparations; a 28% increase in the average labeling efficiency obtained using 111In-oxine; greater than 95% cell viability as measured by the trypan blue exclusion test; elimination of contaminating red blood cells from the leukocyte pellet prior to labeling; and 80% survivability at 15 min post injection (measured as per cent of blood activity on leukocyte fraction).


Yield of trihalomethanes and haloacetic acids upon chlorinating algal cells, and its prediction via algal cellular biochemical composition.
The major objective of the present study was to investigate the contribution of major biomolecules, including protein, carbohydrates and lipids, in predicting DBPs formation upon chlorination of algal cells. Three model compounds, including bovine serum albumin (BSA), starch and fish oil, as surrogates of algal-derived proteins, carbohydrates and lipids, and cells of three algae species, representing blue-green algae, green algae, and diatoms, were chlorinated in the laboratory. The results showed that BSA (27 microg mg(-1) C) and fish oil (50 microg mg(-1) C) produced more than nine times higher levels of chloroform than starch (3 microg mg(-1) C). For the formation of HAAs, BSA was shown to have higher reactivity (49 microg mg(-1) C) than fish oil and starch (5 microg mg(-1) C). For the algal cells, Nitzschia sp. (diatom) showed higher chloroform yields (48 microg mg(-1) C) but lower HAA yields (43 microg mg(-1) C) than Chlamydomonas sp. (green algae) (chloroform: 34 microg mg(-1) C; HAA: 62 microg mg(-1) C) and Oscillatoria sp. (blue-green algae) (chloroform: 26 microg mg(-1) C; HAA: 72 microg mg(-1) C). The calculated chloroform formation of cells from the three algal groups, based on their biochemical compositions, was generally consistent with the experimental data, while the predicted values for HAAs were significantly lower than the observed ones. As compared to humic substances, such as humic and fulvic acids, the algal cells appeared to be important precursors of dichloroacetic acid.


ABCA7 haplodeficiency disturbs microglial immune responses in the mouse brain.
Carrying premature termination codons in 1 allele of the ABCA7 gene is associated with an increased risk for Alzheimer's disease (AD). While the primary function of ABCA7 is to regulate the transport of phospholipids and cholesterol, ABCA7 is also involved in maintaining homeostasis of the immune system. Since inflammatory pathways causatively or consequently participate in AD pathogenesis, we studied the effects of Abca7 haplodeficiency in mice on brain immune responses under acute and chronic conditions. When acute inflammation was induced through peripheral lipopolysaccharide injection in control or heterozygous Abca7 knockout mice, partial ABCA7 deficiency diminished proinflammatory responses by impairing CD14 expression in the brain. On breeding to AppNL-G-F knockin mice, we observed increased amyloid-β (Aβ) accumulation and abnormal endosomal morphology in microglia. Taken together, our results demonstrate that ABCA7 loss of function may contribute to AD pathogenesis by altering proper microglial responses to acute inflammatory challenges and during the development of amyloid pathology, providing insight into disease mechanisms and possible treatment strategies.


Factors of physical activity among Chinese children and adolescents: a systematic review.
Lack of physical activity is a growing problem in China, due to the fast economic development and changing living environment over the past two decades. The aim of this review is to summarize the factors related to physical activity in Chinese children and adolescents during this distinct period of development. A systematic search was finished on Jan 10th, 2017, and identified 2200 hits through PubMed and Web of Science. English-language published studies were included if they reported statistical associations between factors and physical activity. Adapted criteria from the Strengthening The Reporting of OBservational studies in Epidemiology (STROBE) statement and evaluation of the quality of prognosis studies in systematic reviews (QUIPS) were used to assess the risk of bias of the included studies. Related factors that were reported in at least three studies were summarized separately for children and adolescents using a semi-quantitative method. Forty two papers (published 2002-2016) were included. Most designs were cross-sectional (79%), and most studies used questionnaires to assess physical activity. Sample size was above 1000 in 18 papers (43%). Thirty seven studies (88%) showed acceptable quality by methodological quality assessment. Most studies reported a low level of physical activity. Boys were consistently more active than girls, the parental physical activity was positively associated with children and adolescents' physical activity, children in suburban/rural regions showed less activity than in urban regions, and, specifically in adolescents, self-efficacy was positively associated with physical activity. Family socioeconomic status and parental education were not associated with physical activity in children and adolescents. The studies included in this review were large but mostly of low quality in terms of study design (cross-sectional) and methods (questionnaires). Parental physical activity and self-efficacy are promising targets for future physical activity promotion programmes. The low level of physical activity raises concern, especially in suburban/rural regions. Future research is required to enhance our understanding of other influences, such as the physical environment, especially in early childhood.


The Physalis peruviana leaf transcriptome: assembly, annotation and gene model prediction.
Physalis peruviana commonly known as Cape gooseberry is a member of the Solanaceae family that has an increasing popularity due to its nutritional and medicinal values. A broad range of genomic tools is available for other Solanaceae, including tomato and potato. However, limited genomic resources are currently available for Cape gooseberry. We report the generation of a total of 652,614 P. peruviana Expressed Sequence Tags (ESTs), using 454 GS FLX Titanium technology. ESTs, with an average length of 371 bp, were obtained from a normalized leaf cDNA library prepared using a Colombian commercial variety. De novo assembling was performed to generate a collection of 24,014 isotigs and 110,921 singletons, with an average length of 1,638 bp and 354 bp, respectively. Functional annotation was performed using NCBI's BLAST tools and Blast2GO, which identified putative functions for 21,191 assembled sequences, including gene families involved in all the major biological processes and molecular functions as well as defense response and amino acid metabolism pathways. Gene model predictions in P. peruviana were obtained by using the genomes of Solanum lycopersicum (tomato) and Solanum tuberosum (potato). We predict 9,436 P. peruviana sequences with multiple-exon models and conserved intron positions with respect to the potato and tomato genomes. Additionally, to study species diversity we developed 5,971 SSR markers from assembled ESTs. We present the first comprehensive analysis of the Physalis peruviana leaf transcriptome, which will provide valuable resources for development of genetic tools in the species. Assembled transcripts with gene models could serve as potential candidates for marker discovery with a variety of applications including: functional diversity, conservation and improvement to increase productivity and fruit quality. P. peruviana was estimated to be phylogenetically branched out before the divergence of five other Solanaceae family members, S. lycopersicum, S. tuberosum, Capsicum spp, S. melongena and Petunia spp.


Determination of chlorthalidone in plasma, urine and red blood cells by gas chromatography with nitrogen detection.
A sensitive and selective gas chromatographic method is described for determining the diuretic and antihypertensive drug chlorthalidone in plasma, urine and erythrocytes. Use is made of an alkali flame ionization detector (nitrogen detector), and the chlorthalidone and internal standard are chromatographed as methyl derivatives. Down to 10 ng of drugs in the biological sample can be measured accurately, with a standard deviation of 5%. Because the concentration of chlorthalidone found in erythrocytes is 50-100 times higher than that in plasma, the influence of haemolysis on the plasma concentration has been investigated. In addition, a pharmacokinetic study with human volunteers revealed that the apparent concentration of the drug found in plasma can be much too low (by more than 50%), if the plasma is not separated from the erythrocytes immediately after venipuncture. Precautions to be observed to ensure correct handling of blood samples (so that results for plasma concentrations will be reliable) are stressed. The findings have application in kinetic studies on chlorthalidone.


Mechanical failure after locking plate fixation of unstable intertrochanteric femur fractures.
The aim of this study was analyze modes of mechanical failure in a consecutive cohort of patients and establish possible risk factors. This was a retrospective cohort study. The study was conducted at an academic level-1 trauma center. Twenty-nine patients (mean age 56 years, range 21-92; 45% males, 41% smokers, 17% diabetic, mean body mass index 26.9, range 20-56) with 30 OTA 31A3 fractures treated between 2003 and 2007 were included. Operative fixation using 4.5-mm locking compression plate (LCP) proximal femur plate (Synthes, Paoli, PA). Mechanical failure was defined as loss of alignment of at least 10° or shortening of at least 2 cm. Secondary outcomes included patient and fixation construct variables as possible predictors for mechanical failure. At 20 months of follow-up, 11 failures (37%) occurred. Mean time to failure was 18 weeks (range 2-84). Cumulative failure rates were 10%, 20%, 27%, and 33% at 1, 2, 6, and 12 months, respectively. The most frequent failure mode was varus collapse with screw cut out (5 cases). There was no statistically significant difference between groups with regards to age, body mass index, diabetes, or smoking habit. The presence of a "kickstand screw" and medial cortical reduction were not significantly different in cases with and without failure. Proximal screw number and type were similar in both groups. A high rate of mechanical failure can be expected with proximal locking plate fixation of unstable proximal femur fractures. The use of a "kickstand" screw could not be established to reduce the risk for mechanical failure. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


5' end-centered expression profiling using cap-analysis gene expression and next-generation sequencing.
Cap-analysis gene expression (CAGE) provides accurate high-throughput measurement of RNA expression. CAGE allows mapping of all the initiation sites of both capped coding and noncoding RNAs. In addition, transcriptional start sites within promoters are characterized at single-nucleotide resolution. The latter allows the regulatory inputs driving gene expression to be studied, which in turn enables the construction of transcriptional networks. Here we provide an optimized protocol for the construction of CAGE libraries on the basis of the preparation of 27-nt-long tags corresponding to initial bases at the 5' ends of capped RNAs. We have optimized the methods using simple steps based on filtration, which altogether takes 4 d to complete. The CAGE tags can be readily sequenced with Illumina sequencers, and upon modification they are also amenable to sequencing using other platforms.


Virtual Apartment-Based Stroop for assessing distractor inhibition in healthy aging.
Executive functioning declines found in healthy aging may impact negatively on everyday activities. Inhibitory control is an executive function that involves the preclusion of irrelevant information and prohibition of prepotent responses. Decreased inhibitory control has been suggested as a feature of age-related cognitive decline. In this study, we compared a Virtual Apartment Stroop Task with paper-and-pencil and computerized Stroop modalities in older adults and young adults. The primary results were (a) within groups the classic "Stroop pattern" found in traditional modalities (e.g., paper-and-pencil; computer automated) was observed in the Virtual Apartment Stroop Task; (b) the Virtual Apartment "Stroop scores" between groups were consistent with significant differences found between older adults and young adults on the paper-and-pencil and computerized Stroop tasks; and (c) individuals in the older adult cohort performed significantly more poorly on the Virtual Apartment Stroop Task than did participants in the young adult cohort. These results suggest the potential of the Virtual Apartment Stroop Task to distinguish between prepotent response inhibition and resistance to distractor inhibition in aging adults.


Response of Preterm Infants to 2 Noninvasive Ventilatory Support Systems: Nasal CPAP and Nasal Intermittent Positive-Pressure Ventilation.
Noninvasive ventilation (NIV) in preterm infants is currently applied using intermittent positive pressure (2 positive-pressure levels) or in a conventional manner (one pressure level). However, there are no studies in the literature comparing the chances of failure of these NIV methods. The aim of this study was to evaluate the occurrence of failure of 2 noninvasive ventilatory support systems in preterm neonates over a period of 48 h. A randomized, prospective, clinical study was conducted on 80 newborns (gestational age < 37 weeks, birthweight < 2,500 g). The infants were randomized into 2 groups: 40 infants were treated with nasal CPAP and 40 infants with nasal intermittent positive-pressure ventilation (NIPPV). The occurrence of apnea, progression of respiratory distress, nose bleeding, and agitation was defined as ventilation failure. The need for intubation and re-intubation after failure was also observed. There were no significant differences in birth characteristics between groups. Ventilatory support failure was observed in 25 (62.5%) newborns treated with nasal CPAP and in 12 (30%) newborns treated with NIPPV, indicating an association between NIV failure and the absence of intermittent positive pressure (odds ratio [OR] 1.22, P < .05). Apnea (32.5%) was the main reason for nasal CPAP failure. After failure, 25% (OR 0.33) of the newborns receiving nasal CPAP and 12.5% (OR 0.14) receiving NIPPV required invasive mechanical ventilation. Ventilatory support failure was significantly more frequent when nasal CPAP was used.


Activation of single Ca2(+)-dependent K+ channel by external ATP in mouse macrophages.
Single Ca2(+)-dependent K+ currents responding to external ATP were recorded from cell-attached patches on mouse peritoneal macrophages. Extracellularly applied ATP activated an inward single-channel current with a conductance of 25 pS and a reversal potential of -79 mV (pipette potential, Vp) when the pipette contained a 145 mM KCl solution. The reversal potential was shifted 56 mV positive by a 10-fold reduction in external (pipette) K+ concentration. The effect of ATP was abolished by either removal of external Ca2+ or treatment with an intracellular Ca2+ chelator, the acetoxymethyl ester of 1,2-bis (2-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid (BAPTA-AM). This channel has a mean open time of 9.1 ms and open probability was not strongly dependent on Vp in the range tested (+120 to -30 mV).


Enhancement of protein sensitivity for MALDI imaging mass spectrometry after chemical treatment of tissue sections.
MALDI imaging mass spectrometry (IMS) has become a valuable tool for the investigation of the content and distribution of molecular species in tissue specimens. Numerous methodological improvements have been made to optimize tissue section preparation and matrix deposition protocols, as well as MS data acquisition and processing. In particular for proteomic analyses, washing the tissue sections before matrix deposition has proven useful to improve spectral qualities by increasing ion yields and the number of signals observed. We systematically explore here the effects of several solvent combinations for washing tissue sections. To minimize experimental variability, all of the measurements were performed on serial sections cut from a single mouse liver tissue block. Several other key steps of the process such as matrix deposition and MS data acquisition and processing have also been automated or standardized. To assess efficacy, after each washing procedure the total ion current and number of peaks were counted from the resulting protein profiles. These results were correlated to on-tissue measurements obtained for lipids. Using similar approaches, several selected washing procedures were also tested for their ability to extend the lifetime as well as revive previously cut tissue sections. The effects of these washes on automated matrix deposition and crystallization behavior as well as their ability to preserve tissue histology were also studied. Finally, in a full-scale IMS study, these washing procedures were tested on a human renal cell carcinoma biopsy.


Kaempferol and quercetin stimulate granulocyte-macrophage colony-stimulating factor secretion in human prostate cancer cells.
Granulocyte-macrophage colony-stimulating factor (GM-CSF) holds immunotherapeutic promise in prostate cancer as it activates the host immune system. Increased production of GM-CSF by cancer cells may facilitate host immunosurveillence by the dendritic cells (DC). Here, we studied the effects of kaempferol (K) and quercetin (Q) on the production of GM-CSF in PC-3 cells. Human cytokine antibody array revealed that treatment with K or Q increased GM-CSF release by PC-3 cells. We further observed by ELISA that K and Q in a concentration-dependent manner increased GM-CSF production without affecting its mRNA levels. Inhibitors of vesicular traffic through the endoplasmic reticulum and Golgi-blocked GM-CSF secretory stimulation. A microtubule-stabilizing agent stimulated GM-CSF release, whereas tubulin and actin depolymerizers suppressed K- or Q-stimulated secretion of GM-CSF. Depletion of extracellular or intracellular calcium ion inhibited the GM-CSF secretion upregulated by both K and Q. Furthermore, we showed that K- and Q-stimulated GM-CSF production involves PLC, PKC, and MEK1/2 activation. Treating human DC with the conditioned medium of K- or Q-incubated PC-3 cells increased chemotaxis of DC, which was significantly attenuated when the conditioned medium was incubated with the neutralizing antibody against GM-CSF. Taken together, our results demonstrate that K and Q activate an immune response in the prostate cancer cells by stimulating GM-CSF production, which in turn could result in the recruitment of DCs to the tumor site.


Irinotecan: a new antineoplastic agent for the management of colorectal cancer.
To review the pharmacologic, pharmacokinetic, therapeutic, and safety aspects of irinotecan, a new antineoplastic agent, and to assess its role in the treatment of colorectal and lung cancer. English-language articles from the MEDLINE database, January 1990-March 1998; Pharmacia & Upjohn Company; published articles and meeting abstracts. Studies in humans with cancer, clinical case reports, and open clinical studies were reviewed. Efficacy studies were limited to trials with at least 20 evaluable patients. Relevant data were extracted from published reports and abstracts. Irinotecan is an effective agent for the treatment of advanced colorectal cancer. It demonstrates significant activity as a first-line agent and in patients with disease that is refractory to fluorouracil-containing regimens. Activity against lung cancer has also been demonstrated. Limited data indicate activity against cancers of the ovary, cervix, stomach, and in non-Hodgkin's lymphomas. Major toxicity consists of myelosuppression and diarrhea. Irinotecan is a useful addition to the antineoplastic drug family and offers significant efficacy for treatment of patients with fluorouracil-refractory colorectal cancer.


Human papillomavirus vaccination recommendation may be linked to reimbursement: a survey of Virginia family practitioners and gynecologists.
Concerns have been raised about gynecologists as vaccinators. This survey evaluated use of the human papillomavirus (HPV) vaccine, attitudes, and barriers among gynecologists and family practitioners for differences between the 2 specialties. A cross-sectional survey was conducted using a 50-item, self-administered questionnaire mailed to participants. The study was conducted in Virginia through the University of Virginia Center for Survey Research. The questionnaire was mailed to 500 family practitioners and 500 gynecologists. The questionnaire asked provider and practice demographics, vaccine practices, knowledge, HPV vaccine attitudes, and barriers to vaccination. We compared gynecologists to family practitioners for the outcome of offering HPV vaccination. Logistic regression was performed to determine factors associated with providers choosing to offer and recommend the HPV vaccine. After exclusion of ineligible physicians, 385 of 790 doctors responded (48.7%). Seventy percent of family practitioners and 73.5% of gynecologists currently offer the HPV vaccine. There were no significant differences in demographics or practice patterns between the specialties. The most frequent barrier to vaccination reported by both groups was reimbursement. In multivariate logistic regression, inadequate reimbursement was negatively associated with offering the HPV vaccine (odds ratio [OR] 0.41; 95% confidence interval [CI] 0.15-1.1) and with recommending the vaccine to patients (OR 0.45; 95% CI 0.26-0.76). Experience treating HPV- related disease was positively associated with offering the HPV vaccine (OR 2.3; 95% CI 1.1-4.8). Gynecologists are providing HPV vaccination at rates similar to family practitioners. Reimbursement concerns may negatively have an impact on doctors' recommendation of the HPV vaccine.


Neonatal screening for cystic fibrosis is beneficial even in the context of modern treatment.
To determine whether early identification of babies with cystic fibrosis (CF) improves outcome in the current environment of new improved treatments, considering the criticism that there may be only marginal benefit gained by CF newborn screening (NBS). We tested whether CF NBS in the setting of modern CF center care still afforded benefit using the UK CF Database (UKCFD; ) to compare clinical outcomes in infants who underwent NBS and control subjects who were clinically diagnosed (CD). With Mann-Whitney rank tests, 184 patients who underwent NBS aged 1 to 9 years in 2002 (excluding meconium ileus) were compared with matched patients who were CD in 3-year age groups (950 control subjects). Patients as old as 6 years who underwent NBS had significantly greater median height z-scores, less severe Northern chest radiography scores, better Shwachman-Kulczycki scores, and lower rates of chronic Pseudomonas aeruginosa infection. No difference was found for weight z-score or % predicted forced expiratory value in 1 second or forced volume capacity. Nutritional benefit was demonstrated in patients who underwent NBS and were homozygous for the DeltaF508 mutation. NBS segregates with better outcomes in patients as old as 6 years compared with age- and gene-matched control subjects who are CD. This cross-sectional study shows that infants who undergo screening derive nutritional benefit in improved median height and reduced morbidity.


Universal Screening for Familial Hypercholesterolemia in Children.
Individuals with familial hypercholesterolemia (FH) who are untreated have up to 100-fold elevated risk for cardiovascular complications compared with those who are unaffected. Data for identification of FH with a universal screening for hypercholesterolemia in children are lacking. This study sought genetic identification of FH from a cohort of children with elevated serum total cholesterol (TC) concentration, detected in a national universal screening for hypercholesterolemia. Slovenian children born between 1989 and 2009 (n = 272) with TC >6 mmol/l (231.7 mg/dl) or >5 mmol/l (193.1 mg/dl) plus a family history positive for premature cardiovascular complications, identified in a national universal screening for hypercholesterolemia at 5 years of age were genotyped for variants in LDLR, PCSK9, APOB, and APOE. Of the referred children, 57.0% carried disease-causing variants for FH: 38.6% in LDLR, 18.4% in APOB, and none in PCSK9. Nine novel disease-causing variants were identified, 8 in LDLR, and 1 in APOB. Of the remaining participants, 43.6% carried the APOE E4 isoform. Estimated detection rate of FH in the universal screening program from 2009 to 2013 was 53.6% (95% confidence interval [CI]: 34.5% to 72.8%), peaking in 2013 with an upper estimated detection rate of 96.3%. Variants in LDLR, APOB, or the APOE E4 isoform occurred in 48.6%, 60.0%, and 76.5%, respectively, of patients with a family history negative for cardiovascular complications. Most participants who were referred from a national database of universal screening results for hypercholesterolemia had genetically confirmed FH. Data for family history may not suffice for reliable identification of patients through selective and cascade screening.


The role of calcium in the repetitive firing of neostriatal neurons.
The Ca++ -dependence of the repetitive firing of neostriatal neurons was studied in an in vitro slice preparation of the rat neostriatum. Neuronal firing was evoked by injecting depolarizing currents of 100-200 ms duration. In normal conditions, the mode of firing was tonic and showed very little adaptation. The frequency-current relation was linear over a wide range of frequencies. The repetitive firing was first enhanced and later suppressed by Co++, Mn++ and Cd++. These effects on the repetitive firing by the Ca++ -channel blockers paralleled the suppression of the slow afterhyperpolarizing potential. The lowering (0.2 mM) of Ca++ had similar effects. In the presence of TEA (up to 10 mM), the cell fired both Na+ and Ca+ action potentials. The results suggest that, as in other CNS neurons of the vertebrate, in neostriatal neurons the slow afterhyperpolarizing potential (AHP) is due to a Ca++ -activated K+ -conductance, and that the AHP plays a crucial role in the repetitive firing of these neurons.


The Investigation of Rate of Birth Asphyxia and its Relationship with Delivery Mode at Shahid Beheshti Hospital of Isfahan during 2013, 2014, and 2015.
Birth asphyxia is considered as one of the biggest challenges faced by perinatal care experts. According to the WHO, in 2005, one-fourth of infant mortality cases occurred due to birth asphyxia. This study is a retrospective study done on the newborn population with gestational ages of 36 weeks or higher during the years 2013, 2014, and 2015 to find the relationship between the number of birth asphyxia cases and the years. As a secondary objective, the relationship between the mode of delivery and the cases of birth asphyxia were investigated using logistic regression test. The inclusion criteria consisted of three cases, namely, gestational age of 36 weeks or higher, 5 min Apgar of <5, and umbilical cord pH of <7.1. The exclusion criteria involved hydrops fetalis, cyanotic heart disease, chromosomal abnormality, and congenital infections. From the two independent variables of the study, only the variable "year" showed a significant difference between the years 2013 and 2015 (P < 0.01). The delivery mode did not have any statistically significant influence on the newborns' affliction with birth asphyxia (P = 0.993). According to the results, there is a significant difference between the rate of birth asphyxia in Iran and its global rate in 2015, and regarding its multiple growth from 2013 to 2015, conducting a more comprehensive and extensive research on birth asphyxia risk factors at a delivery time seems justified and inevitable.


Sexual, seasonal and tissue variation in the encystment of Cotylurus variegatus metacercariae in perch, Perca fluviatilis.
All of the 267 perch sampled from Lough Neagh between 1981 and 1983 were infected with the metacercarial cysts of Cotylurus variegatus. Sites of infection were the swim-bladder, pericardium, septum transversum and, to a lesser extent, the visceral cavity. The swim-bladder, particularly the anterior portion, was the site of heaviest infection. Visceral cysts were found chiefly in female fish and this may be related to reduced immunological defence and/or thinner body wall during the breeding season. The number of cysts recorded was not related to host length, suggesting that further parasite invasion is offset by cyst mortality possibly as a result of the host immunological response to the parasite. Highest mean worm burden was recorded between May and June. This corresponded to increased water temperature necessary for development of eggs and the breeding season (April-June) of the perch.


The evolving role of advanced MR imaging as a management tool for adult ischemic stroke: a Western-European perspective.
New and more advanced diagnostic imaging techniques for acute stroke triage have the potential to not only improve the quality of care but also reduce health care costs. Although sufficiently large and methodologically sound studies with regard to cost effectiveness of MR imaging are lacking, the overall impression is that MR imaging has revolutionized not only the diagnosis but also the open and investigational management of neurologically ill patients.


Insights into stem cell factor chemotactic guidance of neural crest cells revealed by a real-time directionality-based assay.
The extracellular environment through which neural crest cells (NCCs) translocate and differentiate plays a crucial role in the determination of cell migration and homing. In the trunk, NCC-derived melanocyte precursor cells (MPCs) take the dorsolateral pathway and colonize the skin, where they differentiate into pigment cells (PCs). Our hypothesis was that the skin, the MPCs' target tissue, may induce a directional response of NCCs toward diffusible factor(s). We show that the treatment of in vitro NCCs with skin extract (SE) or Stem Cell Factor (SCF) contributes to maintaining proliferative activity, accelerates melanocyte differentiation, and guides a subpopulation of NCCs by chemotaxis toward the gradient source of these factors, suggesting that they may represent the MPCs' subpopulation. Current data on stimulated directional persistence of NCCs supports the participation of diffusible molecules in the target colonization mechanism, guiding MPCs to migrate and invade the skin. Our results show similar effects of SE and SCF on NCC growth, proliferation and pigment cell differentiation. Also, the use of a proven real-time directionality-based objective assay shows the directional migration of NCCs toward SE and SCF, indicating that the epidermal SCF molecule may be involved in the chemotactic guidance mechanism of in vivo NCCs. Although SCF is the strongest candidate to account for these phenomena, the nature of other factor(s) affecting NCC-oriented migration remains to be investigated. This data amplifies the functional scope of trophic factors by involving them in new cell behaviors such as molecular guidance in the colonization mechanism of embryonic cells.


Low-frequency resonance Raman studies of the H(M202)G cavity mutant of bacterial photosynthetic reaction centers.
Low-frequency (90-435 cm(-1)) NIR-excitation (875-900 nm) resonance Raman (RR) studies are reported for the H(M202)G cavity mutant of bacterial photosynthetic reaction centers (RCs) from Rb. sphaeroides that was first described by Goldsmith et al. [(1996) Biochemistry 35: 2421-2428]. In this mutant, the His residue that axially ligates the Mg ion of the M-side bacteriochlorophyll (BChl) of the special pair primary donor (P) is replaced by a non-ligating Gly residue. Regardless, the Mg ion of P(M) in the H(M202)G RCs remains pentacoordinates and is presumably ligated by a water molecule, although this axial ligand has not been definitively identified. The low-frequency RR studies of the H(M202)G RCs are accompanied by studies of RCs exchanged with D(2)O and incubated with imidazole (Im). The RR studies of the cavity mutant RCs reveal the following: (1) The structure of P(M) in the H(M202)G RCs is different from that of the wild-type, consistent with an altered BChl core. (2) A water ligand for P(M) in the H(M202)G RCs is generally consistent with the low-frequency RR spectra. The Mg-OH(2) stretching vibration is tentatively assigned to a band at 318 cm(-1), a frequency higher than that of the Mg-His stretch of the native pigment ( approximately approximately 235 cm(-1)). (3) The BChl core structure of P(M) in the cavity mutant is rendered similar (but not identical) to that of the wild-type when the adventitious water axial ligand is replaced by Im. (4) Exchange with D(2)O results in more global structural changes, likely involving the protein, which in turn affect the structure of the BChls in P. (5) Assignment of the low-frequency vibrational spectrum of P is generally more complex than originally suggested.


Melanocortin-4 receptors activate sympathetic preganglionic neurons and elevate blood pressure via TRPV1.
Melanocortin-4 receptors (MC4Rs) expressed by the central nervous system are essential regulators of energy homeostasis, and Mc4r mutation is the most common cause of human monogenic obesity. Notably, patients with obesity carrying Mc4r mutations are protected against obesity-induced hypertension, and MC4R agonists elevate blood pressure (BP). Although increased sympathetic tone by MC4Rs is suggested to underlie this phenotype, the detailed mechanisms remain unclear. Here, we investigate how MC4Rs regulate the sympathetic preganglionic neurons and find that MC4Rs activate these neurons via the protein kinase A-dependent activation of the transient receptor potential vanilloid 1 (TRPV1) channel. Importantly, we demonstrate that the inhibition of TRPV1 prevents MC4R-induced elevation of BP but does not affect MC4R-induced anorexia. We further show that TRPV1 is responsible for MC4R-dependent activation of the sympathetic preganglionic neurons by high-fat diet. Together, our results provide insight into how MC4Rs regulate sympathetic function.


The science of anterior teeth selection for a completely edentulous patient: a literature review.
Esthetic replacement and physiological tooth arrangement made the complete denture biologically compatible and desirable. Proper placement of tooth should be functional and esthetically pleasing to enhance the psychology of the patient. This article reviews the evolution of concepts for teeth selection and the recent techniques employed for selecting anterior teeth for complete dentures.


Spotlight on the role of hormonal factors in the emergence of autoreactive B-lymphocytes.
Pathogenic autoimmunity requires a combination of inherited and acquired factors. In as much as hormones influence the sexual dimorphism of the immune system, it is possible that they can initiate or accelerate an autoimmune process, and contribute to gender-biased autoimmune disorders. Not only natural hormones, but also endocrine disruptors, such as environmental estrogens, may act in conjunction with other factors to override immune tolerance to self-antigens. In lupus, murine and human studies demonstrate that female sex hormones are implicated in disease pathogenesis. In the B cell compartment, both prolactin and estrogen are immunomodulators that affect maturation, selection and antibody secretion. Their impact may be based on their capacity to allow autoreactive B cells to escape the normal mechanisms of tolerance and to accumulate in sufficient numbers to cause clinically apparent disease. Both hormones lead to the survival and activation of autoreactive B cells, but they skew B cell maturation towards different directions, with prolactin inducing T cell-dependent autoreactive follicular B cells and estrogen eliciting T cell-independent autoreactive marginal zone B cells. Differential modulation of the cytokine milieu by hormones may also affect the development and activation of specific mature B cell subsets. This novel insight suggests that targeted manipulation of these pathways may represent a promising avenue in the treatment of lupus and other gender-biased autoimmune diseases.


An investigation of the origin, location and variations of the renal arteries in human fetuses and their clinical relevance.
We investigated the origin, localizations and anatomic variations of the renal artery (RA) in human fetuses with the aim of determining the distribution of these variations according to lateralization and gender. In total, 90 fetuses of spontaneous abortion (45 males, 45 females) with no congenital malformations were included to the study. The abdominal aorta and its branches were dissected after latex solution colored with red ink had been injected into the vessels from the thoracic aorta. In all, 180 RA dissections were performed bilaterally in 90 cases and the anatomic variations were photographed. Right and left RAs were found to originate from the following levels according to the columna vertebralis, respectively: 3.8% and 1.9% lower T12, 67.3% and 25.0% upper L1, 9.6% and 28.8% mid L1, 15.3% and 40.3 lower L1, 3.8% and 3.8% upper 1/3 part of L2 vertebra. The right RA originated from the lateral part and anterolateral wall of the abdominal aorta in 73.0% and 26.9% of cases while the lateral and anterolateral wall origin percentages of left RA were 90.3% and 9.6%, respectively. The origin site of the right RA from the abdominal aorta was superior to, at the same level with, and inferior to that of the left RA in 53.8%, 34.6% and 11.5% of the cases, respectively. There were no variations in 75% of the cases whereas the remaining 25% had several variation patterns. The presented morphological results are as follows: A single hilar artery in 75% of the cases, double hilar arteries in 11.1%, an inferior polar artery in 10.5%, and a superior polar artery in 3.3% of specimens studied. Anatomical variations were observed more frequently among male fetuses and on the right side. Knowledge of RA variations is important for surgeons in performing many procedures and may help to avoid clinical complications, especially, during radiological examination and/or surgical approaches in the abdominal region.


Enantioselective Synthesis of Substituted δ-Lactones by Cooperative Oxidative N-Heterocyclic Carbene and Lewis Acid Catalysis.
Efficient construction of complex cylcopentane- or cyclohexane-fused δ-lactones employing redox activation of enals using a chiral N-heterocyclic carbene and LiCl as cooperative catalysts is described. The organocascade proceeds with excellent diastereo- (>99:1) and enantioselectivity (up to >99% ee) and comprises the formation of three bonds with three contiguous stereocenters.


Neutrophils from pregnant women produce thromboxane and tumor necrosis factor-alpha in response to linoleic acid and oxidative stress.
Preeclampsia is associated with oxidative stress, neutrophil activation, neutrophil infiltration into systemic vasculature, and elevated plasma levels of linoleic acid, the fatty acid precursor to arachidonic acid and its metabolite, thromboxane. In this study we evaluated whether linoleic acid under conditions of oxidative stress would stimulate neutrophil production of thromboxane and tumor necrosis factor-alpha. Neutrophils were isolated from 14 normal pregnant women. Western blot demonstrated cyclooxygenase-2 expression at 18 hours of incubation, so this incubation time was used for experiments. Neutrophils (2 x 10(6) cells/mL) were incubated in Dulbecco's modified Eagle's medium/F-12 with: (1) linoleic acid control; (2) an oxidizing solution enriched with linoleic acid; (3) oxidizing solution enriched with linoleic acid plus indomethacin; (4) oxidizing solution enriched with linoleic acid plus aspirin; (5) oxidizing solution enriched with linoleic acid plus NS-398, a specific inhibitor of cyclooxygenase-2; or (6) oxidizing solution enriched with linoleic acid plus pinane thromboxane, a thromboxane synthase inhibitor and receptor blocker. Oxidizing solution enriched with linoleic acid significantly increased oxidative stress in neutrophils. Compared with linoleic acid, oxidizing solution enriched with linoleic acid significantly increased neutrophil production of thromboxane and tumor necrosis factor-alpha. Indomethacin and aspirin inhibited oxidizing solution enriched with linoleic acid stimulation of thromboxane, but NS-398 was equally effective implicating cyclooxygenase-2 in the thromboxane response. Indomethacin inhibited oxidizing solution enriched with linoleic acid stimulation of tumor necrosis factor-alpha, but so did pinane thromboxane implicating thromboxane in the tumor necrosis factor-alpha response. These data demonstrate that exposure of neutrophils from normal pregnant women to conditions present in preeclamptic women results in neutrophil activation with release of thromboxane and tumor necrosis factor-alpha. Newly synthesized thromboxane is cyclooxygenase-2 dependent and plays a role in the tumor necrosis factor-alpha response. Our data suggest a mechanism for maternal vasoconstriction and vascular inflammation in preeclampsia because activated, thromboxane-secreting neutrophils migrate across endothelium into the microenvironment of the vasculature in which they could promote vasoconstriction, whereas release of tumor necrosis factor-alpha could cause vascular inflammation.


Pilonidal sinus: a 5-year appraisal of the Millar-Lord treatment.
One hundred and two patients with a pilonidal sinus treated by minor excision had a life-table of 5-year freedom from recurrence of 89% provided that they attended a dressing clinic until the wound healed. With failure to attend the clinic, the recurrence freedom fell to 57% (P less than 0.004). Age, sex, acute abscess, previous excision, presence of hairs and the number of midline pits and lateral abscesses did not significantly influence the result. The method supersedes more extensive procedures, but should be tested against even less traumatic techniques.


Transcript levels of plastin 3 and neuritin 1 modifier genes in spinal muscular atrophy siblings.
In single gene disorders, patients with the same genotype may have variations in severity. One of the main factors affecting disease severity is modifier genes. Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder caused by degeneration of alpha motor neurons. Plastin 3 (PLS3) is a phenotypic modifier of SMA, and neuritin 1 (NRN1) has also been suggested as a possible modifier gene. The aim of the present study was therefore to analyze PLS3 and NRN1 expression in SMA siblings in four families. The study group consisted of four SMA families with seven with discordant phenotype and two affected siblings. Total RNA was isolated from whole blood. PLS3 and NRN1 expression was analyzed on quantitative real-time polymerase chain reaction. In family 1 only NRN1 expression was increased in the mildly affected sister. In family 2 only PLS3 had a modifier effect. Family 3, which had type III siblings with identical clinical phenotypes, had similar PLS3 expression between the siblings but no NRN1 expression. In family 4, neither PLS3 nor NRN1 had any correlation with severity. On analysis of the expression of NRN1 in SMA patients for the first time, NRN1 could be a potential modifier gene. PLS3 expression does not always modify SMA phenotype. In patients with no modifier effect of known genes, genome sequencing and transcriptome analysis are promising for the identification of novel modifiers and understanding of SMA pathophysiology.


Infection of nonhuman primates with recombinant human metapneumovirus lacking the SH, G, or M2-2 protein categorizes each as a nonessential accessory protein and identifies vaccine candidates.
Recombinant human metapneumovirus (HMPV) in which the SH, G, or M2 gene or open reading frame was deleted by reverse genetics was evaluated for replication and vaccine efficacy following topical administration to the respiratory tract of African green monkeys, a permissive primate host. Replication of the deltaSH virus was only marginally less efficient than that of wild-type HMPV, whereas the deltaG and deltaM2-2 viruses were reduced sixfold and 160-fold in the upper respiratory tract and 3,200-fold and 4,000-fold in the lower respiratory tract, respectively. Even with the highly attenuated mutants, there was unequivocal HMPV replication at each anatomical site in each animal. Thus, none of these three proteins is essential for HMPV replication in a primate host, although G and M2-2 increased the efficiency of replication. Each gene-deletion virus was highly immunogenic and protective against wild-type HMPV challenge. The deltaG and deltaM2-2 viruses are promising vaccine candidates that are based on independent mechanisms of attenuation and are appropriate for clinical evaluation.


Targeting smooth muscle microRNAs for therapeutic benefit in vascular disease.
In view of the bioinformatic projection that a third of all protein coding genes and essentially all biological pathways are under control of microRNAs (miRNAs), it is not surprising that this class of small RNAs plays roles in vascular disease progression. MiRNAs have been shown to be involved in cholesterol turnover, thrombosis, glucose homeostasis and vascular function. Some miRNAs appear to be specific for certain cells, and the role that such cell-specific miRNAs play in vascular disease is only beginning to be appreciated. A notable example is the miR-143/145 cluster which is enriched in mature and highly differentiated smooth muscle cells (SMCs). Here we outline and discuss the recent literature on SMC-expressed miRNAs in major vascular diseases, including atherosclerosis, neointima formation, aortic aneurysm formation, and pulmonary arterial hypertension. Forced expression of miR-145 emerges as a promising strategy for reduction and stabilization of atherosclerotic plaques as well as for reducing neointimal hyperplasia. It is concluded that if obstacles in the form of delivery and untoward effects of antimirs and mimics can be overcome, the outlook for targeting of SMC-specific miRNAs for therapeutic benefit in vascular disease is bright.


Preparation of crosslinked enzyme aggregates (CLEAs) of acid urease with urethanase activity and their application.
An acid urease from Providencia rettgeri JN-B815 was purified via ultrasonication, ethanol precipitation, and DEAE ion-exchange column chromatography. It was found that the enzyme exhibits not only urease activity, but also urethanase activity, which made it possible to reduce EC already existed or would produce and its precursor urea at the same time. Then, crosslinked enzyme aggregates of P. rettgeri urease (PRU-CLEAs) were prepared using genipin as crosslinking agent. The purification process of acid urease, the effects of genipin concentration, and crosslinking time on PRU-CLEAs activity were investigated. The crosslinking was performed at pH 4.5 for 2.5 h, using 0.3% genipin as crosslinking agent, and 0.3 g · L(-1) bovine serum albumin as protein feeder. Using the obtained PRU-CLEAs, the removal rate of urea was up to 9.31 mg · L(-1) · h(-1). The removal rate of urea was still up to 7.56 mg · L(-1) · h(-1) after PRU-CLEAs was re-used for 6 times. When PRU-CLEAs were applied in a batch stirred and membrane reactor, the removal rate of urea in rice wine reached 5.16 mg · L(-1) · h(-1) and the removal rate of EC was 9.21 μg · L(-1) · h(-1). Furthermore, the treatment with PRU-CLEAs revealed no significant change of volatile flavor substances in Chinese rice wine. Thus PRU-CLEAs have great potential in the elimination of EC in Chinese rice wine.


Highly stable carbon nanotube field emitters on small metal tips against electrical arcing.
Carbon nanotube (CNT) field emitters that exhibit extremely high stability against high-voltage arcing have been demonstrated. The CNT emitters were fabricated on a sharp copper tip substrate that produces a high electric field. A metal mixture composed of silver, copper, and indium micro- and nanoparticles was used as a binder to attach CNTs to the substrate. Due to the strong adhesion of the metal mixture, CNTs were not detached from the substrate even after many intense arcing events. Through electrical conditioning of the as-prepared CNT emitters, vertically standing CNTs with almost the same heights were formed on the substrate surface and most of loosely bound impurities were removed from the substrate. Consequently, no arcing was observed during the normal operation of the CNT emitters and the emission current remained constant even after intentionally inducing arcing at current densities up to 70 mA/cm2.


Matrine enhances osteogenic differentiation of bone marrow-derived mesenchymal stem cells and promotes bone regeneration in rapid maxillary expansion.
The aim of this study was to evaluate the influence of matrine on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (BMSCs) as well as on bone metabolism in a rat rapid maxillary expansion (RME) model. In in vitro experiments, rat BMSCs were adopted and cell proliferation of BMSCs was measured. Meanwhile, the osteogenic differentiation of BMSCs was detected by alkaline phosphatase (ALP) activity assay, Alizarin red S staining and gene expression. In vivo bone regeneration was analyzed in a rat RME model. Eighteen rats were divided into 3 groups: one group without any treatment, one group receiving only RME, and a group with RME and matrine treatment. After 2 weeks, new bone formation was detected by Micro-CT and histology. Immunohistochemical staining was used to evaluate ALP and BMP2 expression. Overall, we found that matrine upregulated cell proliferation dose-dependently. Also, ALP activity and mineralized matrix generation were enhanced. Moreover, the osteoblast-related gene expression (ALP, bone sialoprotein and osteocalcin) by BMSCs was also promoted. Micro-CT revealed that matrine significantly promoted in vivo bone formation after 2 weeks. Concomitantly, histological examination of haematoxylin-eosin, safranin-O and toluidine blue staining confirmed these findings. In addition, the levels of ALP and BMP2 in the palatal suture tissues of rats with matrine treatment were the highest among three groups. This work suggests that matrine regulates osteogenesis and enhances bone regeneration. Matrine treatment may be beneficial in improving the stability of maxillary expansion.


Increase of cloning efficiencies by using high-performance liquid chromatography-purified vectors and linkers.
High-performance liquid chromatography (HPLC) was applied to the purification of vector and linker DNAs used in molecular cloning experiments. The data clearly demonstrate that HPLC (Blue Column from LKB/TSK G 4000 SW) based on steric exclusion is a very rapid and convenient method for the size separation of DNA fragments that are virtually free of any contamination. Application of vector and linker DNAs, purified by this method, enabled us to clone nanogram amounts of avian myeloblastosis virus RNA, which was used for the evaluation of a recently described cloning procedure.


Effects of cancer-associated point mutations on the structure, function, and stability of isocitrate dehydrogenase 2.
Mutations in isocitrate dehydrogenase (IDH) are frequently found in low-grade gliomas, secondary glioblastoma, chondrosarcoma, acute myeloid leukemias, and intrahepatic cholangiocarcinoma. However, the molecular mechanisms of how IDH2 mutations induce carcinogenesis remain unclear. Using overlapping PCR, transfection, immunoblotting, immunoprecipitation, measurements of enzyme activity, glucose, lactic acid, ATP, and reactive oxygen species (ROS), cell viability, protein degradation assays post-inhibition of the 26S proteasome (bortezomib) or HSP90 (17-AAG), and a homology model, we demonstrated that the properties of ten cancer-associated IDH2 variants (R140G/Q/W and R172S/K/M/W/G/C/P) arising from point mutations are closely related to their structure and stability. Compared with wild-type IDH2, the R172 and R140 point mutations resulted in a decrease in IDH2 activity, ROS, and lactate levels and an increase in glucose and ATP levels under normal and hypoxic conditions, indicating that mutant IDH2 increases cell dependency on mitochondrial oxidative phosphorylation, and reduces glycolysis under hypoxia. Overexpression of most of IDH2 point mutants showed anti-proliferative effects in the 293T and BV2 cell lines by inhibition of PI3K/AKT signaling and cyclin D1 expression and/or induced the expression of TNF-α and IL-6. Furthermore, bortezomib treatment resulted in dramatic degradation of IDH2 mutants, including R140G, R140Q, R140W, R172S and R172K, whereas it had little impact on the expression of WT and other mutants (R172M, R172W, R172G, R172C and R172P). In addition, targeting HSP90 minimally affected the expression of mutated IDH2 due to a lack of interaction between HSP90 and IDH2. The homology model further revealed that changes in conformation and IDH2 protein stability appeared to be associated with these point mutations. Taken together, our findings provide information important for understanding the molecular mechanisms of IDH2 mutations in tumors.


[Endovascular treatment of extensive carotid cavernous fistulas by superior ophthalmic vein approach. Technique and illustration of two cases].
Treatment of carotid cavernous fistulas is a domain of interventional neuroradiolgy. In rare cases, however, the cavernous sinus cannot be reached by an endovascular approach, leaving the access via the superior ophthalmic vein as the last therapeutic option. History, diagnostic findings, and operative procedure for a patient with spontaneous, indirect CCF (Barrow type D) and a patient with traumatic, direct CCF (Barrow type A) are described. In both patients, curative therapeutic success could be achieved. This article focuses on the detailed description and illustration of the combined surgical/endovascular technique.


Porcine ubiquitin-like protein MNSFβ promotes cell apoptosis and covalently binds to BCL-G to enhance staurosporine-induced apoptosis.
Monoclonal non-specific suppressor factor β (MNSFβ) is a ubiquitously expressed member of the ubiquitin-like family. It functions as a regulator of cell apoptosis and a potential tumor suppressor, playing a vital role in the processes of immune cell function and apoptosis. The present study constructed GFP-pMNSFβ swine umbilical vein endothelial cell (SUVEC) lines and investigated the function of porcine MNSFβ (pMNSFβ) in apoptosis, as well as its interactions with pBCL-G. Results revealed that stably expressed pMNSFβ protein in SUVEC lines significantly enhanced staurosporine (STS)-induced apoptosis. pMNSFβ proteins interacted with pBCL-G proteins and the expression of these interacting proteins synergized to further enhance STS-induced apoptosis. GFP-pMNSFβ stably expressed SUVEC lines through transient transfection and neomycin screening methods. Over 90% of the SUVEC cultures expressed GFP signals, and 41.5 kDa GFP-pMNSFβ proteins were detected with western blotting methods. Annexin V-PE/PI staining and flow cytometry analyses showed that overexpression of pMNSFβ proteins significantly elevated STS-induced apoptosis rates. Co-immunoprecipitation methods revealed an interaction between pMNSFβ and pBCL-G proteins. BCL-G is a proapoptotic member of the BCL-2 family that has been shown to be misexpressed in human systemic lupus erythematosus, as well as mammary and prostate cancers. Here, we demonstrated that the co-expression and potential conjugation of pMNSFβ and pBCL-G proteins synergistically enhanced STS-induced apoptosis. The present study was the first to characterize the function of MNSFβ in porcine cells, and to clarify the function of MNSFβ in apoptosis. These results reveal that pMNSFβ is a potential molecular model for future investigations of diseases related to human MNSFβ dysfunction.


[Epidemiological aspects of salmonelloses in Turin. II: The presence of Salmonella in surface waters and sewerage waters of Turin].
The Authors have examined several sets of superficial water and of sewerage waters (respectively 44 and 27) in the metropolitan area of Turin in search of Salmonella bacteria.