The new CYP3A4 intron 6 C>T polymorphism (CYP3A4*22) is associated with an increased risk of delayed graft function and worse renal function in cyclosporine-treated kidney transplant patients.
Cyclosporine A (CsA) is a substrate of cytochrome P450 3A4 (CYP3A4). Recently, a newly discovered intron 6 single-nucleotide polymorphism in CYP3A4 (rs35599367 C>T), defining the CYP3A4*22 allele, has been linked to reduced hepatic expression and activity of CYP3A4. In the present study, the clinical impact of this single-nucleotide polymorphism was investigated in a cohort of patients receiving a CsA-based immunosuppressive regimen. A total of 172 de-novo kidney transplant recipients, receiving CsA/mycophenolate mofetil as immunosuppressive therapy and participating in the Fixed-Dose Concentration Controlled study, were genotyped for the new CYP3A4*22 allele. CsA C(0) and/or C(2) levels were measured on days 3 and 10 and in months 1, 3, 6, and 12 after transplantation. Plasma creatinine concentrations, delayed graft function (DGF), and biopsy-proven acute rejection were recorded. The CYP3A4*22 allele was significantly associated with a higher risk of DGF compared with the CYP3A4*1/*1 patients after adjustment for known risk factors [odds ratio (OR)=6.34, confidence interval (CI(95%): 1.38-29.3), P=0.015]. Mixed-model analysis demonstrated that the overall creatinine clearance was 20% lower in CYP3A4*22 allele carriers compared with CYP3A4*1/*1 patients [CI(95%) (-33.1 to -7.2%), P=0.002]. For ABCB1 3435C>T, T-variant carriers had a decreased risk of developing DGF compared with CC patients [CT: OR=0.30, CI(95%) (0.11-0.77), P=0.011; TT: OR=0.18, CI(95%) (0.05-0.67), P=0.011]. CYP3A4*22 constitutes a risk factor for DGF and worse creatinine clearance in patients receiving CsA-based immunosuppressive therapy. Therefore, pretransplant genotyping for the CYP3A4*22 allele might help clinicians to identify patients at risk of DGF and poor renal function when treated with CsA.


CMR imaging predicts death and other adverse events in suspected cardiac sarcoidosis.
This study aimed to demonstrate that the presence of late gadolinium enhancement (LGE) is a predictor of death and other adverse events in patients with suspected cardiac sarcoidosis. Cardiac sarcoidosis is the most important cause of patient mortality in systemic sarcoidosis, yielding a 5-year mortality rate between 25% and 66% despite immunosuppressive treatment. Other groups have shown that LGE may hold promise in predicting future adverse events in this patient group. We included 155 consecutive patients with systemic sarcoidosis who underwent cardiac magnetic resonance (CMR) for workup of suspected cardiac sarcoid involvement. The median follow-up time was 2.6 years. Primary endpoints were death, aborted sudden cardiac death, and appropriate implantable cardioverter-defibrillator (ICD) discharge. Secondary endpoints were ventricular tachycardia (VT) and nonsustained VT. LGE was present in 39 patients (25.5%). The presence of LGE yields a Cox hazard ratio (HR) of 31.6 for death, aborted sudden cardiac death, or appropriate ICD discharge, and of 33.9 for any event. This is superior to functional or clinical parameters such as left ventricular (LV) ejection fraction (EF), LV end-diastolic volume, or presentation as heart failure, yielding HRs between 0.99 (per % increase LVEF) and 1.004 (presentation as heart failure), and between 0.94 and 1.2 for potentially lethal or other adverse events, respectively. Except for 1 patient dying from pulmonary infection, no patient without LGE died or experienced any event during follow-up, even if the LV was enlarged and the LVEF severely impaired. Among our population of sarcoid patients with nonspecific symptoms, the presence of myocardial scar indicated by LGE was the best independent predictor of potentially lethal events, as well as other adverse events, yielding a Cox HR of 31.6 and of 33.9, respectively. These data support the necessity for future large, longitudinal follow-up studies to definitely establish LGE as an independent predictor of cardiac death in sarcoidosis, as well as to evaluate the incremental prognostic value of additional parameters.


A Proposed Framework to Identify Dispensable and Essential Functions in Bifidobacteria: Case Study of Bifidobacterium breve UCC2003 as a Prototype of Its Genus.
Functional genomics of bacteria commonly aims at establishing genotype-phenotype links in microorganisms of industrial, technological and biomedical relevance. In this regard, random transposon mutagenesis coupled to high-throughput next-generation sequencing approaches, termed transposon-insertion sequencing (TIS), has emerged as a robust, genome-wide alternative to perform functional genome analysis. Though these approaches have been used in a large number of studies involving pathogenic and clinically relevant bacteria, they have received little attention in the fields of commensal and potentially beneficial bacteria, including probiotic microorganisms. In this chapter, we describe the implementation of the TIS method Transposon-Directed Insertion Sequencing to describe the set of essential genes in a representative strain of a genus encompassing several commensal and potentially probiotic bacteria and discuss considerations when applying similar methodological approaches to other Bifidobacterium species/strains of interest.


Identification and characterization of three novel lipases belonging to families II and V from Anaerovibrio lipolyticus 5ST.
Following the isolation, cultivation and characterization of the rumen bacterium Anaerovibrio lipolyticus in the 1960s, it has been recognized as one of the major species involved in lipid hydrolysis in ruminant animals. However, there has been limited characterization of the lipases from the bacterium, despite the importance of understanding lipolysis and its impact on subsequent biohydrogenation of polyunsaturated fatty acids by rumen microbes. This study describes the draft genome of Anaerovibrio lipolytica 5ST, and the characterization of three lipolytic genes and their translated protein. The uncompleted draft genome was 2.83 Mbp and comprised of 2,673 coding sequences with a G+C content of 43.3%. Three putative lipase genes, alipA, alipB and alipC, encoding 492-, 438- and 248- amino acid peptides respectively, were identified using RAST. Phylogenetic analysis indicated that alipA and alipB clustered with the GDSL/SGNH family II, and alipC clustered with lipolytic enzymes from family V. Subsequent expression and purification of the enzymes showed that they were thermally unstable and had higher activities at neutral to alkaline pH. Substrate specificity assays indicated that the enzymes had higher hydrolytic activity against caprylate (C8), laurate (C12) and myristate (C14).


Experimental observation of microplastics invading the endoderm of anthozoan polyps.
Coral reefs are being degraded worldwide by land reclamation and environmental factors, such as high seawater temperature, resulting in mass bleaching events. In addition, microplastics disturb the formation of coral-algae symbiotic relationships in primary polyps. In our experiments, we observed this effect in the bleached primary polyp Seriatopora caliendrum that lost its symbiont Symbiodiniaceae as a result of high water temperature. There was a higher incorporation of microspheres into bleached corals than in healthy ones. To understand the interference in symbiosis, we used the sea anemone Exaiptasia (as an anthozoan model organism) and fed it with microspheres. TEM results suggested the incorporation of microspheres and symbionts from the same phagocytosis zones in the mesenterial filament and endocytosis by the cells. In the tentacles, microspheres were in the same cell layer as the symbionts. These results suggest that microplastics occupy the spaces inhabited by Symbiodiniaceae, thereby hindering their symbiotic association.


Complex and extensive infective endocarditis: a novel surgical approach.
Endocarditis involving the central fibrous body of the heart requires carefully planned surgical intervention. We present a novel approach in a 65-year-old male with extensive endocarditis involving the aortic root, ventricular septum, central fibrous body together with mitral, aortic and tricuspid valves.


The composition of the gut microbiota is altered in biliary atresia with cholangitis.
To detect the composition of the gut microbiota in biliary atresia after Kasai surgery. Infants within six months after the Kasai operation who were diagnosed by cholangiography at Shanghai Children's Hospital were enrolled in the study. Fecal samples were collected from diapers, placed into sterile tubes in the inpatient department or outpatient department and frozen at -80°C within half an hour. The gut microbiota was detected by 16S rRNA sequences. Then, the patients that were followed up to one year after the Kasai operation who suffered from cholangitis at least one time were grouped into the BAcho group, and the others were grouped into the BAnoncho group. Nine of 18 BA patients were grouped into the BAcho group, and the others were grouped into the BAnoncho group. In the BAcho group, AST, ALT and GGT were significantly increased compared to the BAnoncho group. The number of total OTUs (operational taxonomic units) in feces was more elevated in the BAnoncho group than in the BAcho group. In the BAnoncho group, the Chao index at the OTU level was significantly increased compared to that in the BAcho group (66.37 ± 21.5 vs. 45.64 ± 11.25, p = 0.02 < 0.05). Bifidobacterium was the most abundant genus in the BAnoncho group, accounting for 22.14%, and Klebsiella accounted for 22.74% in the BAcho group. Compared with the BAnoncho group, Bacteroides was significantly decreased in the BAcho group (p = 0.037). The composition of the gut microbiota was different between BA with cholangitis and BA without cholangitis.


Evaluation of stability and antibacterial properties of extracted chlorophyll from alfalfa (Medicago sativa L.).
This study aims to the evaluation of stability and antibacterial properties of the extracted chlorophyll from alfalfa. For this purpose, chlorophylls a and b from alfalfa were extracted by enzymatic and ultrasound methods. The results show that the content of chlorophyll a in alfalfa is higher than chlorophyll b and also the enzymatic method demonstrates higher yield in chlorophyll extraction. In the present study, the chlorophyll stability was evaluated in different conditions including temperature (-18, 4 and 25 °C), time (15, 30 and 45 days), pH (4.5 and 5.5) and NaCl concentration (50, 100 and 150 mM). Also, antibacterial effects were investigated at different concentrations of chlorophyll (20, 40, 60 and 100 μM) against some bactriaes by agar disk diffusion and microdilution (MIC and MBC) methods. The results demonstrate that 50 mM of NaCl, temperature -18 °C, pH = 4.5 and time 15 days are associated with the highest chlorophyll a and b contents. Furthermore, the resistance of bacterias in agar disk diffusion and microdilution methods observe Listeria < Staphylococcus < Salmonella < Escherichia < Pseudomonas and Listeria<(Staphylococcus = Escherichia = Salmonella)<Pseudomonas, respectively. Also, there are significant differences between different chlorophyll concentrations against Listeria and Staphylococcus in evaluation of inhibition effects of total extracted chlorophyll (p < 0.05).


Catalytic ozonation of 2, 2'-methylenebis (4-methyl-6-tert-butylphenol) over nano-Fe3O4@cow dung ash composites: Optimization, toxicity, and degradation mechanisms.
Composite magnetic oxide at cow dung ash, nano-Fe3O4@cow dung ash (nano-Fe3O4@CDA), was used as catalytic material for the degradation of 2, 2'-methylenebis (4-methyl-6-tert-butylphenol) (AO 2246) in real biologically pretreated landfill leachate. The Fe3O4@CDA composite exhibited catalytic ozonation activity and allowed material separation and magnetic recovery. The effects of several operating parameters including O3 concentration, catalyst dosage, temperature and scavengers were evaluated in parallel. Over 70% of AO 2246 were removed by the nano-Fe3O4@CDA/O3 system under optimum conditions within 120min reaction time. The EPR, GC-MS and free-radical quenching experiments expatiated the mechanism of this degradation process. It was confirmed that the AO 2246 was degraded efficiently in this catalytic micro-ozonation process, Additionally, GC-MS analysis state clearly that the 3,5-bis(1,1-dimethylethyl)phenol, 4-(1,5-dihydroxy-2,6,6-trimethylcyclohex-2-enyl)but-3-en-2-one, ethanone, 1-(1,4-dimethyl-3-cyclohexen-1-yl)-, 5-tert-butyl-6-3, 5-diene-2-one, 2-hydroxyhexanoic acid, 2-propenoic acid 1,1-dimethylethyl ester, butanoic acid, 2-methyl-, methyl ester and propanoic acid, 2, 2-dimethyl- were the dominant oxidation products (OPs) during the degradation of the AO 2246. The EPR results showed that the catalytic ozonation over Fe3O4@CDA led to produce more hydroxyl radicals, which were in favor of AO 2246 degradation. The toxicity evolution was also performed through a QSAR analysis calculated by the ECOSAR program which further demonstrated the different responses toward the AO 2246 and its OPs.


Correction: Reusable manganese compounds containing pyrazole-based ligands for olefin epoxidation reactions.
Correction for 'Reusable manganese compounds containing pyrazole-based ligands for olefin epoxidation reactions' by Ester Manrique et al., Dalton Trans., 2015, 44, 17529-17543.


Simulation training in the obstetrics and gynecology clerkship.
The purpose of this study was to determine the effectiveness of obstetrics simulator training for medical students by comparing measures of confidence in normal obstetrics skills of students with and without training. After a lecture on normal labor and delivery, 33 third-year students practiced their skills either on an obstetrics simulator (n = 17) or received no further formal instruction (n = 16). All students were asked to respond to surveys of their experience and confidence in performing obstetrics procedures. Students who practiced deliveries on the simulator were more likely to believe that they could perform most portions of a vaginal delivery with minimal supervision or independently than were students who did not receive simulator experience. Fifteen students (88%) who received simulator experience felt that they were ready to attempt a vaginal delivery independently or with minimal supervision compared with 2 students (12.5%) who received a lecture only (P < .001). Students who practiced deliveries on an obstetrics simulator report higher levels of confidence in their skills to perform vaginal deliveries.


The exon/intron organization of the globin gene of Scapharca inaequivalvis homodimeric hemoglobin: unusual intron homology with other bivalve mollusc globin genes.
In this study, we have investigated the positions of introns in the globin gene of Scapharca inaequivalvis homodimeric hemoglobin. We found the three exon/two intron organization typical of vertebrate globin genes, with the two introns in highly conserved positions, as it occurs in the A and B globin genes of the tetrameric hemoglobin from the same organism, confirming the absence of the so-called 'central intron' found in the globin genes of plants and of some invertebrates. We identified two homodimeric globin genes (3207 and 2723 bp) that differ only with respect to the size of the first intron. Sequence analysis of the two first introns (1668 and 1364 bp) has revealed that they are highly homologous, except for a 569- and 296-bp insertion in each intron I. Interestingly, the two first introns contain regions with an unusually high identity (approximately 80%) with regions of the first intron of the congeneric clam Anadara trapezia and the related clam Barbatia reveana globin genes, suggesting that these uncoding regions may have played a regulatory role that has subsequently been lost during the course of the evolution.


The complex role of sleep in adolescent depression.
Psychological and pharmacologic treatments for youth depression yield post-acute response and remission rates that are modest at best. Improving these outcomes is an important long-term goal. The authors examine the possibility that a youth cognitive behavioral therapy insomnia intervention may be an adjunct to traditional depression-focused treatment with the aim of improving depression outcomes. This "indirect route" to improving youth depression treatment outcomes is based on research indicating that the risk of depression is increased by primary insomnia and that sleep problems interfere with depression treatment success and on emerging adult depression randomized controlled trial results. The authors describe the protocol they developed.


Structural brain connectivity in children after neonatal stroke: A whole-brain fixel-based analysis.
Neonatal arterial ischemic stroke (NAIS) has been shown to affect white matter (WM) microstructure beyond the lesion. Here, we employed fixel-based analysis, a technique which allows to model and interpret WM alterations in complex arrangements such as crossing fibers, to further characterize the long-term effects of NAIS on the entire WM outside the primary infarct area. 32 children (mean age 7.3 years (SD 0.4), 19 male) with middle cerebral artery NAIS (18 left hemisphere, 14 right hemisphere) and 31 healthy controls (mean age 7.7 years (SD 0.6), 16 male) underwent diffusion MRI scans and clinical examination for manual dexterity. Microstructural and macrostructural properties of the WM were investigated in a fixel-based whole-brain analysis, which allows to detect fiber-specific effects. Additionally, tract-averaged fixel metrics in interhemispheric tracts, and their correlation with manual dexterity, were examined. Significantly reduced microstructural properties were identified, located within the parietal and temporal WM of the affected hemisphere, as well as within their interhemispheric connecting tracts. Tract-averaged fixel metrics showed moderate, significant correlation with manual dexterity of the affected hand. No increased fixel metrics or contralesional alterations were observed. Our results show that NAIS leads to long-term alterations in WM microstructure distant from the lesion site, both within the parietal and temporal lobes as well as in their interhemispheric connections. The functional significance of these findings is demonstrated by the correlations with manual dexterity. The localization of alterations in structures highly connected to the lesioned areas shift our perception of NAIS from a focal towards a developmental network injury.


Peripheral membrane proteins modulate stress tolerance by safeguarding cellulose synthases.
Controlled primary cell wall remodeling allows plant growth under stressful conditions, but how these changes are conveyed to adjust cellulose synthesis is not understood. Here, we identify the TETRATRICOPEPTIDE THIOREDOXIN-LIKE (TTL) proteins as new members of the cellulose synthase complex (CSC) and describe their unique and hitherto unknown dynamic association with the CSC under cellulose-deficient conditions. We find that TTLs are essential for maintaining cellulose synthesis under high-salinity conditions, establishing a stress-resilient cortical microtubule array, and stabilizing CSCs at the plasma membrane. To fulfill these functions, TTLs interact with CELLULOSE SYNTHASE 1 (CESA1) and engage with cortical microtubules to promote their polymerization. We propose that TTLs function as bridges connecting stress perception with dynamic regulation of cellulose biosynthesis at the plasma membrane.


Micelles by self-assembling peptide-conjugate amphiphile: synthesis and structural characterization.
The chemical synthesis by solid-phase methods of a novel amphiphilic peptide, peptide-conjugate amphiphile (PCA), containing in the same molecule three different functions: (i) the N,N-bis[2-[bis(carboxy-ethyl)amino]ethyl]-L-glutamic acid (DTPAGlu) chelating agent, (ii) the CCK8 bioactive peptide, and (iii) a hydrophobic moiety containing four alkyl chains with 18 carbon atoms each, is reported. In water solution at pH 7.4, PCA self-assembles in very stable micelles at very low concentration [critical micellar concentration (cmc) values of 5 x 10(-7) mol kg(-1)] as confirmed by fluorescence spectroscopy. The structural characterization, obtained with small-angle neutron scattering (SANS) measurements, indicates that the aggregates are substantially represented by ellipsoidal micelles with an aggregation number of 39 +/- 2 and the two micellar axes of about 52 and 26 A.


Differential sensitivity for viewpoint between familiar and unfamiliar faces in human visual cortex.
People are extremely proficient at recognizing faces that are familiar to them, but are poor at identifying unfamiliar faces. We used fMR-adaptation to ask whether this difference in recognition might be reflected in the relative viewpoint-dependence of face-selective regions in the brain. A reduced response (adaptation) to repeated images of unfamiliar or familiar faces was found in the fusiform face area (FFA), but not in the superior temporal sulcus (STS) face-selective region. To establish if the neural representation of faces was invariant to changes in viewpoint, we parametrically varied the viewing angle of successive images using 3-dimensional models of unfamiliar and familiar faces. We found adaptation to familiar faces across all changes in viewpoint in the FFA. In contrast, a release from adaptation was apparent in the FFA when unfamiliar faces were viewed at increasing viewing angles. These results provide a neural basis for differences in the recognition of familiar and unfamiliar faces.


Quantification of sugar phosphate intermediates of the pentose phosphate pathway by LC-MS/MS: application to two new inherited defects of metabolism.
We describe a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to quantify pentose phosphate pathway intermediates (triose-3-phosphates, tetrose-4-phosphate, pentose-5-phosphate, pentulose-5-phosphates, hexose-6-phosphates and sedoheptulose-7-phosphate (sed-7P)) in bloodspots, fibroblasts and lymphoblasts. Liquid chromatography was performed using an ion pair loaded C(18) HPLC column and detection of the sugar phosphates was carried out by tandem mass spectrometry using an electron ion spray source operating in the negative mode and multiple reaction monitoring. Reference values for the pentose phosphate pathway intermediates in blood spots, fibroblasts and lymphoblasts were established. The method was applied to cells from patients affected with a deficiency of transaldolase. The transaldolase-deficient cells showed an increased concentration of sedoheptulose-7-phosphate. (Bloodspots: 5.19 and 5.43 micromol/L [0.49-3.33 micromol/L]; fibroblasts 7.43 and 26.46 micromol/mg protein [0.31-1.14 micromol/mg protein]; lymphoblasts 16.03 micromol/mg protein [0.61-2.09 micromol/mg protein].) The method was also applied to study enzymes of the pentose phosphate pathway by incubating fibroblasts or lymphoblasts homogenates with ribose-5-phosphate or 6-phosphogluconate and the subsequent analysis of the formed sugar phosphates.


Relationship between essential fatty acid requirements of aquatic animals and the capacity for bioconversion of linolenic acid to highly unsaturated fatty acids.
1. [1-14C]linolenic acid was injected into the rainbow trout, Salmo gairdnerii, ayu, Plecoglossus altivelis, eel, Anguilla japonica, red sea bream, Chrysophrys major, rockfish, Sebastiscus marmoratus, globefish, Fugu rubripes rubripes and prawn, Penaeus japonicus (molting stage D"1-D2), and the bioconversion of linolenic acid (18:3 omega 3) to highly unsaturated fatty acids such as eicosapentaenoic (20:5 omega 3) and docosahexaenoic (22:6 omega 3) acids was investigated. 2. Linolenic acid was converted to 20:5 omega 3 and 22:6 omega 3 intensively in the rainbow trout, moderately in the ayu, eel and prawn, but slightly in the red sea bream, rockfish and globefish. 3. These results were discussed in relation to the essential fatty acid requirements of the aquatic animals.


JOINTLESS suppresses sympodial identity in inflorescence meristems of tomato.
Unlike monopodial plants, in which flowering terminates growth of a shoot, plants exhibiting sympodial shoot architecture maintain the potential for indeterminate growth even after converting to floral development. This vegetative indeterminacy is conferred by a special type of axillary meristem, the sympodial meristem, which exhibits precocious but determinate growth. The reiterative formation of sympodial meristems as the plant grows results in a shoot composed of a series of modules, each consisting of a limited number of vegetative nodes and terminated by a flower or inflorescence. To determine how sympodial meristems differ from other shoot meristems, we examined interactions between mutations that affect various shoot meristem types in tomato (Lycopersicon esculentum Mill.). Analysis of double mutant combinations of jointless, lateral suppressor, self-pruning, blind, and anantha showed that sympodial meristems share regulatory features with inflorescence meristems. Genetic studies on the jointless mutation implicated this gene in suppressing sympodial meristem fate in the inflorescence. As this mutation has a second phenotype, the elimination of the pedicel abscission zone, we examined the expression pattern of JOINTLESS to test whether pedicel development is involved in directing shoot architecture. We found that this MADS box gene is expressed in a variety of shoot meristems, including inflorescence, floral, sympodial, and axillary meristems, as well as in early staged floral organs, in sporogenous tissues of anthers, and in ovules. Lack of expression in developing pedicels indicates abscission zone development does not rely on JOINTLESS transcription in the differentiating cells. We conclude that the primary role of JOINTLESS is to suppress sympodial meristem identity in inflorescence meristems.


Digitalization in pharmaceutical industry: What to focus on under the digital implementation process?
Digitalization of any manufacture industry is a key step in any progress of the production process. The process of digitalization includes both increased use of robotics, automatization solutions and computerization, thereby allowing to reduce costs, to improve efficiency and productivity, and to be flexible to changes. Pharmaceutical Industry (PI) has however been resistant to digitalization, mainly due to fair experience and complexity of the entailed development and manufacture processes. Nevertheless, there is a clear need to digitalize PI as the demand in both traditional and new drugs is constantly growing. Contract Development Manufacture Organizations (CDMOs) have a special digitalizing challenge. Digitalization of PI, and CDMO precisely, should be tightly related to the main aspects of Good Manufacture Practice (GMP), and, to succeed in PI digitalizing requires constant focus on GMP. Close collaboration with constantly changing stakeholders is another important factor which should be in focus during digitalization of CDMO. This paper represents an overview over the main aspects of CDMO digitalization and discusses both the opportunities and challenges of the process, focusing on the practical solutions for successive digital implementation.


Development of a low-dose monthly injectable contraceptive system: II. Pharmacokinetic and pharmacodynamic studies.
A drug delivery system which provides a sustained release of norethindrone (NET) and mestranol (ME) for one month after a single intramuscular injection was assessed as a long-acting injectable contraceptive. The system is based upon well defined particle size crystals of the synthetic steroids maintained in suspension with saline solution. Eight healthy ovulating women volunteered for the study; they received a combination of 10 mg of NET plus 1 mg of ME in 1 ml of vehicle by intramuscular injection on day five of their menstrual cycle. Blood samples were drawn at 0, 1, 5, 10, 13, 17 and 21 days after drug administration. The immunoreactive serum levels of estradiol, progesterone, NET and ethinylestradiol were measured by specific radioimmunoassay procedures to assess ovarian function and the kinetic parameters of the synthetic steroids. This newly developed contraceptive system proved to be both effective, and long-lasting as well as devoid of side effects.


Evaluation of a photodiode array detector for the verification of peak-homogeneity in high-performance liquid chromatography.
Photodiode array liquid chromatography detectors are claimed to have the ability of evaluating the homogeneity of chromatographic peaks and this could provide a very powerful tool in support of method development. However, in pharmaceutical analysis, for this to be of practical value it must be capable of detecting inhomogeneities at low levels. In this paper, a test has been devised to challenge the sensitivity of instruments to this application. The test makes use of mixtures of the similar benzodiazepines temazepam and lormetazepam in a chromatographic system which does not separate them. One instrument has demonstrated the ability to detect levels of just 0.5% w/w of one benzodiazepine in the other. Statistical F-tests and t-tests have been used to demonstrate that non-homogeneities have been detected with a high level of confidence. It is concluded that photodiode array detectors have the potential to evaluate the homogeneity of chromatographic peaks with a high degree of sensitivity. However, most instruments do not realize this potential because their software does not make proper use of all the data available.


Frailty as a predictor of clinical problems and events that require elderly patients with heart failure to use health resources.
The clinical management of elderly patients with heart failure (HF) is not firmly established. Decision-making should be individualized depending on the biological deterioration of each patient, from aggressive management to a palliative approach. Frailty can serve as the basis for this comprehensive individualized management. Our objective was to evaluate the importance of the main clinical problems, as well as the events that required the use of health resources, based the degree of frailty, in elderly patients with HF. Retrospective observational cohort study. Frailty was defined according to the deficit accumulation construct. A total of 546 patients hospitalized for acute HF were included. The median age (Q1-Q3) was 82 (78-86) years. A total of 454 patients (83%) showed some degree of frailty: 221 (48.7%) mild, 207 (45.6%) moderate and 26 (5.7%) advanced. There was a significant tendency towards polypharmacy from no to severe frailty. Hospital events were recorded for 4 (1-6) patients with mild frailty, 4 (2-6) patients with moderate frailty and 2 ((1-4) patients with advanced frailty (p = 0.045). A total of 204 patients (37.4%) died during follow-up. The median time to death was 11.4 (4-16.8), 6.7 (3.3-11.6), 6.5 (3.4-12.2) and 4.1 (0.8-7.7) months for patients with no, mild, moderate, or advanced frailty, respectively (p = 0.006). Frailty due to deficit accumulation is a good predictor of clinical problems and events that require the use of health resources; therefore, it can serve as a basis for the management of HF in the elderly.


A chiral electrochemical sensor for propranolol based on multi-walled carbon nanotubes/ionic liquids nanocomposite.
A novel chiral electrochemical sensor based on multi-walled carbon nanotubes (MWCNTs)/ionic liquids (ILs) nanocomposite was developed and applied to the enantiomeric recognition of propranolol (PRO). Mechanisms for chiral sensing were discussed in terms of the formation of an efficient chiral nanospace originating from the weak interactions between quasi-chiral MWCNTs and versatile ILs. This work provided new evidence that the electro-oxidation of PRO should result from its hydroxyl group. Herein the ratio of oxidation peak currents, instead of the commonest shift of peak potential, was used to sense the enantiomeric fraction (EF) of (R)-PRO with a linear correlation coefficient of 0.9936. This simple and reliable sensor was successfully applied in an accurate determination of the enantiomeric purity of reagent, as well as the evaluation of waste water treatment efficiency.


Genetic analysis of TMPRSS3 gene in the Korean population with autosomal recessive nonsyndromic hearing loss.
The TMPRSS3 gene (DFNB8/10), which encodes a transmembrane serine protease, is a common hearing loss gene in several populations. Accurate functions of TMPRSS3 in the hearing pathway are still unknown, but TMPRSS3 has been reported to play a crucial role in inner ear development or maintenance. To date, 16 pathogenic mutations have been identified in many countries, but no mutational studies of the TMPRSS3 gene have been conducted in the Korean hearing loss population. In this study, we performed genetic analysis of TMPRSS3 in 40 unrelated Korean patients with autosomal recessive hearing loss to identify the aspect and frequency of TMPRSS3 gene mutations in the Korean population. A total of 22 variations were detected, including a novel variant (p.V291L) and a previously reported pathogenic mutation (p.A306T). The p.A306T mutation which has been detected in only compound heterozygous state in previous studies was identified in homozygous state for the first time in this study. Moreover, the clinical evaluation identified bilateral dilated vestibules in the patient with p.A306T mutation, and it suggested that p.A306T mutation of the TMPRSS3 gene might be associated with vestibular anomalies. In conclusion, this study investigated that only 2.5% of patients with autosomal recessive hearing loss were related to TMPRSS3 mutations suggesting low prevalence of TMPRSS3 gene in Korean hearing loss population. Also, it will provide the information of genotype-phenotype correlation to understand definite role of TMPRSS3 in the auditory system.


Cyclic nucleotide signaling is required during synaptic refinement at the Drosophila neuromuscular junction.
The removal of miswired synapses is a fundamental prerequisite for normal circuit development, leading to clinical problems when aberrant. However, the underlying activity-dependent molecular mechanisms involved in synaptic pruning remain incompletely resolved. Here the dynamic properties of intracellular calcium oscillations and a role for cAMP signaling during synaptic refinement in intact Drosophila embryos were examined using optogenetic tools. We provide In vivo evidence at the single gene level that the calcium-dependent adenylyl cyclase rutabaga, the phosphodiesterase dunce, the kinase PKA, and Protein Phosphatase 1 (PP1) all operate within a functional signaling pathway to modulate Sema2a-dependent chemorepulsion. It was found that presynaptic cAMP levels were required to be dynamically maintained at an optimal level to suppress connectivity defects. It was also proposed that PP1 may serve as a molecular link between cAMP signaling and CaMKII in the pathway underlying refinement. The results introduced an in vivo model where presynaptic cAMP levels, downstream of electrical activity and calcium influx, act via PKA and PP1 to modulate the neuron's response to chemorepulsion involved in the withdrawal of off-target synaptic contacts. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 77: 39-60, 2017.


Primary Antibiotic Resistance of Helicobacter pylori in China.
Antibiotic resistance is the most important factor leading to the failure of eradication regimens; thus, it is important to obtain regional antibiotic resistance information. This review focuses on the prevalence of Helicobacter pylori primary resistance to clarithromycin, metronidazole, amoxicillin, levofloxacin, tetracycline, and furazolidone in China. We searched the PubMed, EMBASE, the China National Knowledge Infrastructure, and Chinese Biomedical databases from the earliest date of each database to October 2016. The search terms included the following: H. pylori, antibiotic (including clarithromycin, metronidazole, amoxicillin, levofloxacin, tetracycline, and furazolidone) resistance with or without China or different regions of China. The data analysis was performed using MedCalc 15.2.2. Each article was weighted according to the number of isolated H. pylori strains. A pooled proportion analysis was performed. Twenty-three studies (14 studies in English and 9 in Chinese) were included in this review. A total of 6274, 6418, 3921, 5468, 2802, and 275 H. pylori strains were included in this review to evaluate the prevalence of H. pylori primary resistance to clarithromycin, metronidazole, levofloxacin, amoxicillin, tetracycline, and furazolidone, respectively. Overall, the primary resistance rates of clarithromycin, metronidazole, levofloxacin, amoxicillin, tetracycline, and furazolidone were 28.9, 63.8, 28.0, 3.1, 3.9, and 1.7%, respectively. In China, the prevalence of H. pylori primary resistance to clarithromycin, metronidazole, and levofloxacin was high and increased over time, whereas the resistance rates to amoxicillin, tetracycline, and furazolidone were low and stable over time.


Urinary Amino-PAHs in relation to diesel engine emissions and urinary mutagenicity.
Diesel exhaust has long been of health concern due to established toxicity including carcinogenicity in humans. However, the precise components of diesel engine emissions that drive carcinogenesis are still unclear. Limited work has suggested that nitrated polycyclic aromatic hydrocarbons (NPAHs) such as 1-nitropyrene and 2-nitrofluorene may be more abundant in diesel exhaust. The present study aimed to examine whether urinary amino metabolites of these NPAHs were associated with high levels of diesel engine emissions and urinary mutagenicity in a group of highly exposed workers including both smokers and nonsmokers. Spot urine samples were collected immediately following a standard work shift from each of the 54 diesel engine testers and 55 non-tester controls for the analysis of five amino metabolites of NPAHs, and cotinine (a biomarker of tobacco smoke exposure) using liquid chromatography-mass spectrometry. An overnight urine sample was collected in a subgroup of non-smoking participants for mutagenicity analysis using strain YG1041 in the Salmonella (Ames) mutagenicity assay. Personal exposure to fine particles (PM2.5) and more-diesel-specific constituents (elemental carbon and soot) was assessed for the engine testers by measuring breathing-zone concentrations repeatedly over several full work shifts. Results showed that it was 12.8 times more likely to detect 1-aminopyrene and 2.9 times more likely to detect 2-aminofluorene in the engine testers than in unexposed controls. Urinary concentrations of 1-aminopyrene were significantly higher in engine testers (p < 0.001), and strongly correlated with soot and elemental carbon exposure as well as mutagenicity tested in strain YG1041 with metabolic activation (p < 0.001). Smoking did not affect 1-aminopyrene concentrations and 1-aminopyrene relationships with diesel exposure. In contrast, both engine emissions and smoking affected 2-aminofluorene concentrations. The results confirm that urinary 1-aminopyrene may serve as an exposure biomarker for diesel engine emissions and associated mutagenicity.


Synthesis of potential 99mTc nitrido tumor imaging disposition in mice.
Several cationic or neutral technetium-nitrido complexes of Schiff bases [for which 5-methyl 3-(2-hydroxyphenyl methylene) dithiocarbazate (L1) was a prototype], bis-aminoethanethiol (BAT-TM) and macrocyclic amines were prepared. We report here, the synthesis and isolation of these TcNLn complexes from reaction mixtures by high pressure liquid chromatography or sephadex G25 column. In vivo tissue distribution studies were performed on tumor bearing mice (B16, EMT6, 3LL) after i.v. injection of 10-20 microCi of TcNLn. Although pharmacokinetic differences appeared between the three studied ranges, we did not obtain proof of any particular specificity to tumor tissues. Nevertheless, the complexes were very stable and some of the ligands could be used as chelators after linking side chains or groups inducing specific tumor localization.


Impurity profile study of lopinavir and validation of HPLC method for the determination of related substances in lopinavir drug substance.
Several related substances (RS4-RS10) were detected in lopinavir drug substance at levels ranging from 0.03% to 0.1% by employing gradient RP-HPLC. The related substances were identified by LC-MS analysis. These related substances were isolated and characterized by Mass, (1)H NMR and FT-IR spectral data. The separation was achieved on a YMC Pack ODS-AQ (250 mm x 4.6 mm, 5 microm) column thermostated at 45 degrees C using 0.02 M KH(2)PO(4) (pH 2.5): acetonitrile as a mobile phase in gradient elution mode. A PDA detector set at 210 nm was used for detection. The investigated validation elements showed the method has acceptable specificity, accuracy, linearity, precision, robustness and high sensitivity with detection limits and quantitation limits ranging from 0.028 microg/ml to 0.063 microg/ml and 0.084 microg/ml to 0.192 microg/ml respectively. The method can be used for routine quality control analysis and stability testing of lopinavir drug substance.


Nitric oxide synthase inhibition by haem oxygenase decreases macrophage nitric-oxide-dependent cytotoxicity: a negative feedback mechanism for the regulation of nitric oxide production.
Nitric oxide (NO) production in macrophages by inducible nitric oxide synthase (NOS2) has multiple tissue damaging effects and is involved in the pathogenesis of inflammation and graft rejection. Haem oxygenase (HmOx) is the enzyme which degrades haem. Its inducible isoform, HmOx1, was recently shown to increase cellular resistance against oxidative stress and to decrease inflammation and graft rejection. Since haem is an essential cofactor for NOS2 activity, we investigated the effects of HmOx1-induction upon NO secretion in macrophages. We induced HmOx1 in BALB/c bone-marrow-derived macrophages by short-term exposure to haemin (20 micromol/l, 30 min); then we incubated them for 24 h to allow maximal expression of HmOx1 activity. Next, we activated the macrophages with lipopolysaccharide (LPS) and measured their NO production and their NO-dependent cytotoxicity against P815 cells. We found that HmOx induction 24 h before LPS activation in mouse macrophages suppresses their production of NO, while HmOx inhibition (with zinc protoporphyrin) increases NO secretion. NOS2 inhibition is reflected by the decrease of macrophage NO-dependent cytotoxicity against the P815 targets. We therefore propose that HmOx1 is a physiological inhibitor of NOS2 in activated macrophages because it decreases haem availability for NOS2 synthesis. NOS2 inhibition may explain the antinflammatory effects of HmOx induction which could also be used therapeutically in situations when NO hyperproduction leads to cytotoxic effects such as inflammation or transplant rejection.


[Expression of Fas ligand in urogenital malignant cell lines and renal cell carcinoma].
To explore the expression of Fas ligand in urogenital malignant cell lines and renal cell carcinoma. With immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR), we detected the expression of Fas ligand (FasL) in 6 urogenital malignant cell lines, bladder carcinoma cell lines (T(24), EJ, BIU-87), renal cell carcinoma cell lines (GRC-1, RCC-949), prostatic carcinoma cell line (PC-3M), and 10 cases of renal cell carcinoma. The expression of FasL was detected in 3 (BIU-87, RCC-949, GRC-1) of the 6 urogenital malignant cell lines with immunocytochemistry, and 4 (BIU-87, RCC-949, GRC-1, PC-3M) of 6 urogenital malignant cell lines with RT-PCR method. No expression of FasL was detected in bladder carcinoma cell lines T(24) and EJ. Eight of 10 renal cell carcinoma tissues positively expressed FasL mRNA. FasL can express in urogenital malignant cell lines and renal cell carcinoma.


A trafficking checkpoint controls GABA(B) receptor heterodimerization.
Surface expression of GABA(B) receptors requires heterodimerization of GB1 and GB2 subunits, but little is known about mechanisms that ensure efficient heterodimer assembly. We found that expression of the GB1 subunit on the cell surface is prevented through a C-terminal retention motif RXR(R); this sequence is reminiscent of the ER retention/retrieval motif RKR identified in subunits of the ATP-sensitive K+ channel. Interaction of GB1 and GB2 through their C-terminal coiled-coil alpha helices masks the retention signal in GB1, allowing the plasma membrane expression of the assembled complexes. Because individual GABA(B) receptor subunits and improperly assembled receptor complexes are not functional even if expressed on the cell surface, we conclude that a trafficking checkpoint ensures efficient assembly of functional GABA(B) receptors.


A new animal model for evaluation of long-term growth rate over one month by rhGH/PLGA microcapsule formulations.
A new animal model to evaluate the long-term growth rate produced by a sustained-release formulation of recombinant human growth hormone (rhGH) over one month was developed and the usefulness of our microcapsule formulations was demonstrated in this model. Long-term pharmacological effects by subcutaneous injection of microcapsules for sustained release of rhGH were evaluated in hypophysectomized (Hpx) rats treated with immunosuppressive agent along with hormone supplement. Copoly(DL-lactic/glycolic)acid (PLGA) microcapsules for sustained release of rhGH, a two-week sustained-release formulation (rhGH-SR-2W) and a one-month sustained-release formulation (rhGH-SR-1M), were prepared by a solid-in-oil-in-water emulsion solvent evaporation technique. Body-weight gain, body-length gain and serum levels of rat insulin-like growth factor-I (rIGF-I) induced by subcutaneous injection of rhGH-SR were compared with those by daily injections of rhGH solution in Hpx rats for 35 days. Serum IGF-I levels in Hpx rats after the injection of rhGH-SR2W microcapsules were higher than those after daily injections of rhGH solution. Body-length gain, a new parameter, after single injection of rhGH-SR-1M microcapsules demonstrated the higher growth rate than that after daily injections of rhGH solution for 35 days. Thus, single injection of rhGH-SR microcapsules demonstrated long-term pharmacological effects greater than those by daily injections of rhGH solution in a newly developed model, immunosuppressed Hpx rats.


Cryo-EM structure of a native, fully glycosylated, cleaved HIV-1 envelope trimer.
The envelope glycoprotein trimer (Env) on the surface of HIV-1 recognizes CD4(+) T cells and mediates viral entry. During this process, Env undergoes substantial conformational rearrangements, making it difficult to study in its native state. Soluble stabilized trimers have provided valuable insights into the Env structure, but they lack the hydrophobic membrane proximal external region (MPER, an important target of broadly neutralizing antibodies), the transmembrane domain, and the cytoplasmic tail. Here we present (i) a cryogenic electron microscopy (cryo-EM) structure of a clade B virus Env, which lacks only the cytoplasmic tail and is stabilized by the broadly neutralizing antibody PGT151, at a resolution of 4.2 angstroms and (ii) a reconstruction of this form of Env in complex with PGT151 and MPER-targeting antibody 10E8 at a resolution of 8.8 angstroms. These structures provide new insights into the wild-type Env structure.


Suitability of dried olive pulp in slow-growing broilers: performance, meat quality, oxidation products, and intestinal mucosa features.
To assess the effect of dietary dried olive pulp (DOP) on growth performance, meat traits and oxidation, and intestinal mucosa features, a total of 180 male slow-growing broiler chickens (Hubbard) were divided into 3 groups and fed 3 isonitrogenous and isoenergetic diets from 14 d of age until slaughter (49 d). The treatments varied according to 3 DOP levels: a control diet without DOP (DOP0, 0%) and 2 test diets containing 5 and 10% of DOP (DOP5 and DOP10, respectively). Duodenal morphometric indices were measured at the end of the feeding period and included: villus height, crypt depth, villus-to-crypt ratio, and villus surface area. Dietary DOP had no adverse effect on growth performance, dressing percentage, or breast yield of broilers. The breast muscle pH at 24 h was significantly higher in birds fed DOP10 diet compared to those on DOP0 and DOP5 diets. Meat color was also affected by dietary treatments. Feeding DOP did not influence breast meat fatty acid composition, whereas meat from DOP-fed broilers resulted less susceptible to lipid and protein oxidation compared to control diet. Including DOP up to 10% in diet resulted in higher duodenal villus height, crypt depth, and villus height to crypt depth ratio as well as villus surface area. Based on our findings, dietary DOP supported productive traits of slow-growing broilers preserving meat from oxidation and improving intestinal morphometric features. As a result, the current study assessed that olive by-product can be used in broiler ration, resulting in a valuable ingredient as replacement for conventional feeds, which could reduce feeding costs due to the low cost of the olive by-product. Thus, using olive by-products as poultry feed may become economically feasible for producers where the olive oil industries play an important economic role.


Effect of reducing the n-6/n-3 fatty acid ratio on the maternal and fetal leptin axis in relation to infant body composition.
To investigate the effect of reducing the n-6/n-3 fatty acid ratio in maternal nutrition on the maternal and cord blood leptin axis and their association with infant body composition up to 2 years. 208 healthy pregnant women were randomized to either a dietary intervention to reduce the n-6/n-3 fatty acid ratio from 15th week of gestation until 4 months postpartum or a control group. Leptin, soluble leptin receptor and free leptin index were determined in maternal and cord plasma and related to infant body composition assessed by skinfold thicknesses up to 2 years. The intervention had no effect on either the maternal or fetal leptin axis. Maternal leptin in late pregnancy was inversely related to infant weight and lean body mass (LBM) up to 2 years, after multiple adjustments. Cord leptin was positively related to weight, body fat, and LBM at birth, and inversely associated with weight, BMI, fat mass, and LBM at 2 years and weight gain up to 2 years. The contribution of cord leptin to infant outcomes was overall stronger compared with maternal leptin. Both, maternal and fetal leptin were associated with subsequent infant anthropometry with a greater impact of fetal leptin.


Identification of serum biomarkers for necrotizing enterocolitis using aptamer-based proteomics.
Necrotizing enterocolitis (NEC) is a potentially fatal intestinal disease primarily affecting preterm infants. Early diagnosis of neonates with NEC is crucial to improving outcomes; however, traditional diagnostic tools remain inadequate. Biomarkers represent an opportunity to improve the speed and accuracy of diagnosis, but they are not routinely used in clinical practice. In this study, we utilized an aptamer-based proteomic discovery assay to identify new serum biomarkers of NEC. We compared levels of serum proteins in neonates with and without NEC and identified ten differentially expressed serum proteins between these groups. We detected two proteins, C-C motif chemokine ligand 16 (CCL16) and immunoglobulin heavy constant alpha 1 and 2 heterodimer (IGHA1 IGHA2), that were significantly increased during NEC and eight that were significantly decreased. Generation of receiver operating characteristic (ROC) curves revealed that alpha-fetoprotein (AUC = 0.926), glucagon (AUC = 0.860), and IGHA1 IGHA2 (AUC = 0.826) were the proteins that best differentiated patients with and without NEC. These findings indicate that further investigation into these serum proteins as a biomarker for NEC is warranted. In the future, laboratory tests incorporating these differentially expressed proteins may improve the ability of clinicians to diagnose infants with NEC rapidly and accurately.


Anthracene cycloadducts as highly selective chiral auxiliaries.
A new chiral auxiliary was designed and easily prepared from a Diels-Alder cycloadduct of an enantiomerically pure anthracene with maleimide. Excellent diastereoselectivities in Diels-Alder reactions, conjugate additions, and aldol reactions employing these auxiliaries are now reported.


Multicentre comparison between slow-release furosemide and bendroflumethiazide in congestive heart failure.
The effects of slow-release furosemide and bendroflumethiazide in the ambulatory treatment of mild to moderate congestive heart failure were compared in a controlled multicentre study using a randomized, double-blind procedure. In the first trial, slow-release furosemide 60 mg daily was compared with 2.5 mg bendroflumethiazide; in the second trial the daily doses were 30 mg and 2.5 mg, respectively. There were 47 patients in Trial I and 39 in Trial II. The randomised allocation to the treatment regimen was made after a four-week period on bendroflumethiazide 2.5 mg daily, and treatment was continued for 12 weeks. Clinical and laboratory data were recorded before, during and at the end of the trial. Adverse effects were few and minor in both medication groups, and there were no significant intergroup differences. It was concluded slow-release furosemide 30 mg or 60 mg daily was as effective as bendroflumethiazide 2.5 mg daily for maintenance treatment in mild to moderate congestive heart failure.


Monolayer behavior of silica particles at air/water interface: a comparison between chemical and physical modifications of surface.
Silica particles are hydrophobized either by chemical graft of alkyl chains or by physical adsorption of cationic surfactants, alkyltrimethylammonium bromide. The effects of the two modification methods on the monolayer behavior of silica particles at the air/water interface are studied, as well as the packing structure of the particulate films. The results show that the hydrophobicity of particles chemically modified by octanol (SiO2-C8) and dodecanol (SiO2-C12) are similar and higher than that modified by butanol (SiO2-C4). The monolayer composed of particles with higher hydrophobicity shows a large lift-off area, higher compressibility, and significant hysteresis due to the higher particle-particle interaction. As a result, the particulate films exhibit 2-dimensional (2D) aggregative domains of closely-packed structure, but with particle free regions presenting among the domains. The monolayer prepared by SiO2-C4 shows a contrary behavior resulted from the higher particle-water interaction. The particles modified by adsorption of cationic surfactants have an amphiphilic property at the air/water interface. Such monolayer exhibits lower compressibility and hysteresis, higher re-spreading characteristic, and a lower collapse pressure compared with those of the chemically modified particles. A particulate film with high uniformity and closely-packed structure can be obtained by using the octyltrimethylammonium bromide (OTAB) modified particles. When the alkyl chain of surfactant increases, the packing of the particles becomes looser. Such phenomenon is probably caused from the higher probability for the long-chain surfactants to stay at the air/water interface which obstructs the intimate contact of particles.


Radiotherapy of prostate carcinoma: results of treatment and complications.
During October 1974 and December 1981, 204 patients with a localized prostate carcinoma were treated by radiotherapy. One hundred and eighty two patients were suitable for this evaluation. The minimal time of follow-up was 30 months, the mean time 54 months. All patients had a split-course radiotherapy starting by equally weighted opposed pelvic fields with daily doses between 1.8 and 2 Gy, a rest period of 6-7 days and a total dose between 60 and 70 Gy. The overall survival rates (actuarial) for all patients were 64% after 5 years and 55% after 8 years. Several prognostic factors were evaluated. The main factor was tumor grading, the 5 year survival rates fall off from 81% for G1, 65% for G2 to 40% for G3. Split according to T-categories, 5 year survival rates were 73% in T1, 74% in T2, 59% in T3, and 50% in T4. When using the American staging system, we found 75% in stage B and 59% in stage C. There was no statistically significant influence of age and dose on survival, but a clear trend for better survival with higher doses. 36.8% of all patients relapsed, 79% of them developed only distant metastases. Comparing grading and appearance of distant metastases, we found 58% in the group of G3 tumors. There was no influence of transurethral resection (TUR) on survival. Side effects were observed frequently, but mostly mild and transient, late effects were rare.


DNA strand transfer reactions catalyzed by vaccinia topoisomerase I.
Vaccinia virus DNA topoisomerase I forms a 3'-phosphoryl intermediate with duplex DNAs containing the conserved binding/cleavage motif 5'CCCTT decreases. Covalently bound enzyme is capable of transferring the incised DNA strand to a heterologous DNA acceptor containing a 5'OH terminus. Both intramolecular and intermolecular religation reactions are catalyzed. Intramolecular strand transfer occurs to the noncleaved strand of the DNA duplex and results in formation of a hairpin loop. Intermolecular religation to an exogenous DNA strand is favored over hairpin formation and requires the potential for base pairing between the acceptor and the noncleaved strand of the donor complex. As few as 4 potential base pairs are sufficient to support intermolecular transfer. These results in vitro are consistent with the proposal that vaccinia topoisomerase can catalyze sequence-specific strand transfer during genetic recombination in vivo (Shuman, S. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 10104-10108.).


Alcohol and cognitive performance: a longitudinal study of older Japanese Americans. The Kame Project.
Recent data demonstrate that moderate consumption of alcohol (13-52 grams of ethanol per day) may be beneficial to cognitive functioning among older adults. Longitudinal growth curve analyses controlling for baseline age, body mass index (BMI), education/income, migrant status, smoking, history of diagnosed stroke, hypertension, coronary heart disease (CHD), depression, diabetes and stroke (time-varying) were used to examine the relationship between alcohol consumption, gender and cognitive performance over an 8-year follow-up period. The sample included 1624 Japanese American community-dwelling adults aged 65 and older who were cognitively intact at baseline and participated in at least one follow-up examination. Cognitive performance was measured using the Cognitive Abilities Screening Instrument (CASI; 0-100 point scale), a global test of cognitive function. Current consumers (n = 480) scored significantly (p < 0.05) higher on CASI (mean rate of change-1.22 CASI units) over the 8-year follow-up period than past consumers or abstainers (n = 1144; mean rate of change-3.77 CASI units). There was no significant main effect for gender, or an alcohol and gender interaction. This study provides further support regarding the beneficial effects of moderate alcohol consumption on cognitive performance over time. Observed benefits were not modified by gender. Future studies need to determine whether alcohol preserves cognition directly or whether other factors such as physiology or cultural drinking practices are driving the observed association.


Importance of histamine H2 receptors in restraint-morphine interactions.
The effects of the brain-penetrating H2 antagonist zolantidine (ZOL, 3 mg/kg, s.c.) were studied on morphine (MOR, 4 mg/kg, s.c.) antinociception (tail flick test) in the presence and absence of previous restraint stress. Animals were handled for 3 days (to reduce handling stress), restrained for 1 hr or handled on day 4, and tested 24 hrs later. As found previously, restraint enhanced the intensity and duration of MOR antinociception. ZOL potentiated MOR antinociception in handled, non-restrained animals, but inhibited MOR action in restrained animals. In contrast, ZOL had no effects on nociceptive responses in either handled or stressed subjects in the absence of MOR. The data suggest that, in the absence of restraint, brain HA acts at the H2 receptor to inhibit MOR antinociception. In contrast, when an animal has been previously restrained, HA enhances MOR antinociception. Thus, brain HA appears to mediate the restraint-induced potentiation of MOR antinociception. Taken with previous results, the present findings suggest that in the presence of MOR, brain HA can provide bidirectional modulation of nociception. The direction of the modulation seems to depend upon the stress experience of the animal.


Mass changes in myoinositol trisphosphate in human platelets stimulated by thrombin. Inhibitory effects of phorbol ester.
Myoinositol trisphosphate (IP3) is formed when phosphatidylinositol 4,5-bisphosphate (PIP2) is hydrolyzed by phospholipase C. At micromolar concentrations, IP3 is a stimulus for Ca2+ release in both platelet membranes and various permeabilized cells. We have utilized a combination of ion exchange and capillary gas chromatography to quantitate the mass of IP3 produced by human platelets stimulated by thrombin. Accumulations of IP3 are transient and detectable within 5 s of exposure to thrombin. Within 15 s, thrombin (1 unit/ml) promotes the formation of 134 pmol of IP3/10(9) platelets, the equivalent of an intracellular concentration of 13.4 microM. Incubation of platelets with a stimulus for protein kinase C, 12-O-tetradecanoyl phorbol 13-acetate, prior to the addition of thrombin impairs the hydrolysis of PIP2 and the increase in IP3, with 50% inhibition occurring at 60 nM TPA. We conclude that platelets produce sufficient quantities of IP3 to cause Ca2+ release from membrane stores. TPA inhibits the activation of phospholipase C and consequently the generation of IP3. The decreased accumulation of IP3 in platelets exposed to TPA may account for the inhibited rise in cytoplasmic Ca2+ which has been observed in such platelets.


Yak milk promotes renal calcium reabsorption in mice with osteoporosis via the regulation of TRPV5.
The Ca2+-selective epithelial channel TRPV5 plays a significant role in renal calcium reabsorption and improving osteoporosis (OP). In this study, we investigated the mechanisms of yak milk on osteoporosis mice in TRPV5-mediated Ca2+ reabsorption in the kidney. We observed that treatment of OP mice with yak milk reconstructed bone homeostasis demonstrated by increasing the levels of OPG as well as decreasing the levels of TRAP and ALP in serum. Additionally, yak milk reduced the level of parathyroid hormone (PTH) and elevated 1,25-(OH)2D3 and calcitonin (CT), and inhibited the excretion of Ca/Cr and Pi/Cr in OP mice, which explained by regulating hormone levels and thus enhance the renal Ca2+ reabsorption. Further analysis exhibited that yak milk upregulated the expression of TRPV5 protein and mRNA as well as calbindin-D28k in OP mice kidneys. Overall, these outcomes demonstrate that yak milk enhances renal Ca2+ reabsorption through the TRPV5 pathway synergistically with calbindin-D28k, thus ameliorating OP mice. This provides a new perspective for yak milk as a nutritional supplement to prevent osteoporosis.


New potent 5-nitrofuryl derivatives as inhibitors of Trypanosoma cruzi growth. 3D-QSAR (CoMFA) studies.
Growth inhibitory activity in vitro of sixteen new 5-nitrofuryl derivatives against the protozoan parasite Trypanosoma cruzi, the causative agent of American trypanosomiasis, was studied. The designed compounds combine in the same molecule the recognized 5-nitrofuryl group, an oxidative stress promoter, and lateral chains that could interact with biomolecules such as trypanothione reductase. Some of the derivatives were found to be very active against the epimastigote form of the parasite, being near to 3.0-fold more active than the reference compound, nifurtimox. Moreover, three-dimensional requirements for activity were clearly observed using a 3D-QSAR study based on a comparative molecular field analysis (CoMFA). The best CoMFA model, r(2) = 0.970 and q(2) = 0.725, points to the importance of a specific hydrogen-bonding pattern around the carbonyl or thiocarbonyl moieties, as well as the requirement for hydrophobic lateral chains. Theoretical pharmacokinetics (Lipinski's rule, PSA) supports further in vivo studies.


Deciphering the origin of million-fold reactivity observed for the open core diiron [HO-FeIII-O-FeIV[double bond, length as m-dash]O]2+ species towards C-H bond activation: role of spin-states, spin-coupling, and spin-cooperation.
High-valent metal-oxo species have been characterised as key intermediates in both heme and non-heme enzymes that are found to perform efficient aliphatic hydroxylation, epoxidation, halogenation, and dehydrogenation reactions. Several biomimetic model complexes have been synthesised over the years to mimic both the structure and function of metalloenzymes. The diamond-core [Fe2(μ-O)2] is one of the celebrated models in this context as this has been proposed as the catalytically active species in soluble methane monooxygenase enzymes (sMMO), which perform the challenging chemical conversion of methane to methanol at ease. In this context, a report of open core [HO(L)FeIII-O-FeIV(O)(L)]2+ (1) gains attention as this activates C-H bonds a million-fold faster compared to the diamond-core structure and has the dual catalytic ability to perform hydroxylation as well as desaturation with organic substrates. In this study, we have employed density functional methods to probe the origin of the very high reactivity observed for this complex and also to shed light on how this complex performs efficient hydroxylation and desaturation of alkanes. By modelling fifteen possible spin-states for 1 that could potentially participate in the reaction mechanism, our calculations reveal a doublet ground state for 1 arising from antiferromagnetic coupling between the quartet FeIV centre and the sextet FeIII centre, which regulates the reactivity of this species. The unusual stabilisation of the high-spin ground state for FeIV[double bond, length as m-dash]O is due to the strong overlap of with the orbital, reducing the antibonding interactions via spin-cooperation. The electronic structure features computed for 1 are consistent with experiments offering confidence in the methodology chosen. Further, we have probed various mechanistic pathways for the C-H bond activation as well as -OH rebound/desaturation of alkanes. An extremely small barrier height computed for the first hydrogen atom abstraction by the terminal FeIV[double bond, length as m-dash]O unit was found to be responsible for the million-fold activation observed in the experiments. The barrier height computed for -OH rebound by the FeIII-OH unit is also smaller suggesting a facile hydroxylation of organic substrates by 1. A strong spin-cooperation between the two iron centres also reduces the barrier for second hydrogen atom abstraction, thus making the desaturation pathway competitive. Both the spin-state as well as spin-coupling between the two metal centres play a crucial role in dictating the reactivity for species 1. By exploring various mechanistic pathways, our study unveils the fact that the bridged μ-oxo group is a poor electrophile for both C-H activation as well for -OH rebound. As more and more evidence is gathered in recent years for the open core geometry of sMMO enzymes, the idea of enhancing the reactivity via an open-core motif has far-reaching consequences.


Bulimia: disturbed patterns of solitude.
This research suggests that daily solitude has a central role in bulimia. A sample of 15 bulimic patients and 24 normal controls carried electronic pagers for one week and filled out self-reports on their experience in response to randomly timed signals. The bulimics reported the lowest mood states when alone at home, the context in which their symptomatic behavior usually takes place. Further, those who reported the worst experiences in this context were the one's showing the most severe behavioral and affective manifestations of the disorder.


A coumarin hydrazone appended rotatable phenolic scaffold as fluorescent chemosensor for Ag+ ions and its bio imaging applications.
A coumarin hydrazone-phenol conjugate, COH4 was designed, synthesized and utilized for the cation sensing studies by fluorimetry studies. The synthesized chemosensor was completely characterized by the usual spectroscopic and analytical studies. The COH4 receptor was examined for the detection of metal ions, in which it had a noticeable blue shifted fluorescence enhancement for Ag+ ions. Upon binding towards Ag+ ions, the photoinduced electron transfer (PET) process is inhibited via intramolecular charge transfer (ICT) process assisted by the arrest of the carbon-carbon single bond rotation. The binding stoichiometry of COH4 + Ag+ complexation ratio is noted to be 1:2, which was further confirmed by jobs plot method. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.41 µM and 0.13 µM respectively. Moreover, COH4 was successfully utilized for the practical applications of Ag+ ion detection in bacterial cell lines.


Hydrothermal liquefaction of Fe-impregnated water hyacinth for generation of liquid bio-fuels and nano Fe carbon hybrids.
In this work, hydrothermal liquefaction experiments of iron impregnated water hyacinth were performed with a motive to enhance bio-oil yields along with generation of nanometal carbon hybrids. Iron nanoparticles were impregnated and its metal loading was determined by ICP-MS. The impact of operating parameters like temperature, biomass to water ratio and reaction time on bio-oil yields was studied. During hydrothermal liquefaction a maximum total bio-oil yield of 38.1% was obtained at 280 °C along with formation of nanometal carbon hybrids. The light oil and heavy oil fractions were characterized by GCMS and NMR for determining the key components. The light oil mainly comprises of alkanes, alcohols and esters whereas heavy oil contains esters, ethers, carboxylic acids and phenols. XRD and XPS of Fe-impregnated water hyacinth and residues confirmed the transition of Fe+3/+2 to Fe0. TEM analysis resulted an average particle size of Fe nanoparticles around 19.6 nm.


Extraction of thromboxane B2 from urine using an immobilized antibody column for subsequent analysis by gas chromatography-mass spectrometry.
This paper describes an antibody affinity (immunoaffinity) column which, in one step, extracts and sufficiently purifies urinary thromboxane B2 (TXB2) for quantitative analysis by high resolution gas chromatography-negative ion chemical ionization-selected ion monitoring-mass spectrometry (HRGC-NICI-SIM-MS). Polyclonal TXB2 antibody from rabbit was partially purified using immobilized Staphylococcus aureus Protein A. The purified IgG fraction was then immobilized using an N-hydroxysuccimidyl silica gel. The resulting matrix bound 570 ng TXB2 per ml of gel. TXB2 was quantitatively eluted with acetonitrile-water (19:1). Columns constructed from the gel could be used repeatedly since binding capacity was reconstituted using 0.01 M phosphate buffer (pH 7.4) with no apparent loss of activity. Using these columns, urinary TXB2 was sufficiently purified in one step such that in subsequent analysis by HRGC-NICI-SIM-MS interference free chromatograms were observed.


Bacteremia in the chronic spinal cord injury population: risk factors for mortality.
Individuals with spinal cord injury (SCI) have a lifelong increased risk of systemic infection, which may be associated with episodes of life-threatening bacteremia. Information concerning specific organisms causing bacteremia, the sites of primary infection, and clinical predictors for mortality are necessary to provide optimal treatment. A retrospective review of positive blood cultures collected over a 32-month period in chronic SCI patients treated at the Veterans Affairs Medical Center SCI Unit. One hundred and twenty-three episodes of bacteremia occurred in 63 patients during 83 hospitalizations; 30 patients had multiple episodes of bacteremia. There were 1,644 admissions during this period, yielding an incidence of bacteremia of 7.5% (5.8% after excluding positive cultures that were believed to be caused by contaminants). The patients (31 with paraplegia and 32 with quadriplegia) had a mean age of 59 +/- 2 years, and a mean duration of injury of 23 +/- 2 years. Bladder management technique consisted of indwelling bladder catheter (n = 53), ileal conduit (n = 6), intermittent catheterization (n = 2), and spontaneous voiding (n = 2). Episodes of bacteremia were nosocomial in 89 out of 123 episodes. Multiple debilitating factors were present, including pressure ulcers in 36 out of 63 patients, chronic ventilator dependency in 5 out of 63 patients, recent surgical procedures in 17 out of 63 patients, underlying malignancy in 5 out of 63 patients, and evidence of malnutrition in 29 out of 63 patients (serum albumin concentration < 2.5 g/dL). Early mortality rate (death within 30 days of bacteremia) occurred in 8 out of 63 patients (13%) and late mortality (> 1 month following a bacteremic episode) occurred in 10 additional participants, such that total mortality was 1 8 out of 63 (29%). The sources of bacteremia were urinary tract infection (n = 39), presumed contaminant (n = 28), decubitus ulcers (n = 21), intravascular catheter (n = 19), pneumonia (n = 5), and other (n = 11). Gram-negative rods accounted for 26 out of 39 episodes of bacteremia from a urinary source. Methicillin-resistant Staphylococcus aureus, methicillin-sensitive S aureus, and coagulase-negative staphylococci were the predominant organisms when intravascular catheters or pressure ulcers were the source of bacteremia. In this population, bacteremia predominantly was caused by hospital-associated organisms, and occurred mainly in malnourished patients who required hospitalization for major underlying debilitating conditions, particularly pressure ulcers. Chronic indwelling bladder catheters and chronic vascular catheter usage also were highly prevalent in patients with bacteremic episodes. Hypoalbuminemia was the strongest independent predictor for mortality.


Effect of polar interactions on the nonlinear behavior of phenol and aniline in reversed phase liquid chromatography.
The effect of the coverage density and the activity of the unreacted silanol groups of the non-endcapped octadecyl bonded phase on the adsorption properties were investigated. The adsorption of two polar low molecular weight compounds with weak acidic (phenol) and with basic (aniline) character was measured. Adsorption data were acquired by frontal analysis from methanol-water and acetonitrile-water solutions to investigate the influence of the type of the organic modifier on the solute retention mechanism and adsorption under non-linear conditions. The adsorption behavior of phenol changes from Langmuir type (from aqueous mixture of methanol) to BET type (from aqueous mixture of acetonitrile). The adsorption of aniline becomes BET-type, regardless of the mobile phase composition in the tested range of the surface coverage of octadecyl ligands. The nature of the organic modifier significantly affects the retention mechanism and the shape of the overloaded elution bands of the studied compounds.


Poly[[hexa-aqua-(μ(2)-oxalato-κO,O:O,O)bis(μ(3)-pyridine-2,4-dicarboxyl-ato-κN,O:O:O)dicerium(III)] monohydrate].
In the polymeric title compound, {[Ce(2)(C(7)H(3)NO(4))(2)(C(2)O(4))(H(2)O)(6)]·H(2)O}(n), the Ce(3+) cation is nine-coordinated in a distorted CeNO(8) tricapped trigonal-prismatic geometry, formed by three pyridine-2,4-dicarboxyl-ate anions, one oxalate anion and three water mol-ecules. The mid-point of the oxalate anion is located on an inversion center. The oxalate and pyridine-2,4-dicarboxyl-ate anions bridge the Ce(3+) cations, forming a two-dimensional polymeric complex parallel to (010). Inter-molecular classical O-H⋯O hydrogen bonding and weak C-H⋯O hydrogen bonding are present in the crystal structure and π-π stacking [centroid-centroid distance = 3.558 (2) Å] is observed between parallel pyridine rings of adjacent mol-ecules. The uncoordinated water mol-ecule shows an occupancy of 0.5.


Basic aspects of the biochemical reactivity of 4-hydroxynonenal.
4-hydroxynonenal (HNE), a major lipid peroxidation product of n-6 polyunsaturated fatty acids, which was discovered by the late Hermann Esterbauer, is a remarkable trifunctional molecule. Both the hydroxy group and the conjugated system consisting of a C=C double bond and a carbonyl group contribute to the high reactivity of HNE. Most of the biochemical effects of HNE can be explained by its rapid reactions with thiol and amino groups. Among the primary reactants for HNE are the amino acids cysteine, histidine and lysine, which--either free or protein-bound--undergo readily Michael additions to the C=C bond. After this primary reaction, which confers rotational freedom to the C2-C3 bond, secondary reactions may occur involving the carbonyl and the hydroxy group. Primary amines may alternatively react with the carbonyl group to form Schiff bases. Reactions which do not fit into this scheme are the oxidation and the reduction respective of the carbonyl group and the epoxidation of the C=C double bond. Examples will be presented for the interaction of HNE with various classes of biomolecules such as proteins and peptides, lipids and nucleic acids and the biochemical consequences will be discussed.


Glycoprotein biosynthesis by human normal platelets.
Incorporation of radioactive Man, Gal, Fuc, Glc-N, and NANA into washed human normal platelets and endogenous glycoproteins has been found. Both parameters were time dependent. Analysis of hydrolyzed labeled glycoproteins by paper chromatography revealed that the radioactive monosaccharide incubated with the platelets had not been converted into other sugars. Acid hydrolysis demonstrates the presence of a glycosidic linkage. All the effort directed to the demonstration of the existence of a lipid-sugar intermediate in intact human platelets yielded negative results for Man and Glc-N used as precursors. The incorporation of these sugars into glycoproteins is insensitive to bacitracin, suggesting no involvement of lipid-linked saccharides in the synthesis of glycoproteins in human blood platelets. The absence of inhibition of the glycosylation process in the presence of cycloheximide suggests that the sugars are added to proteins present in the intact platelets. These results support the contention that glycoprotein biosynthesis in human blood platelets observed under our experimental conditions is effected through direct sugar nucleotide glycosylation.


Gene expression in the developing rat mandible: a gene array study.
To analyse the molecular events that occur in the developing mandible, gene arrays containing probes for 1176 known genes were used. Total RNA was extracted from the mandibles of 1- and 6-day-old rats. Radiolabeled probes were then synthesised and used to probe the DNA arrays. Of the 1176 genes examined, 306 were detectable, and the latter were analyzed by bioinformatics algorithms. K-means clustering grouped 72 genes into Up, 56 genes into Down and 178 genes into NO change. A large number of genes related to cell receptors (by ligands) were grouped into the Down cluster. Gene array technology appears to be a useful tool for studying the complex process of mandibular development.


Ultrasonic-assisted extraction for phenolic compounds and antioxidant activity of Moroccan Retama sphaerocarpa L. leaves: Simultaneous optimization by response surface methodology and characterization by HPLC/ESI-MS analysis.
This study was designed to optimize the ultrasound-assisted extraction of phenolic compounds and the antioxidant activity of Moroccan Retama sphaerocarpa extracts using response surface methodology (RSM). A central composite design has been conducted to investigate the effects of three factors: extraction period (X1), solvent concentration (X2), and solvent-to-material ratio (X3) on extraction yield, total phenolic content (TPC), flavonoids content (TFC), and antioxidant activity. The obtained results showed that the experimental values agreed with the predicted ones, confirming the capacity of the used model for optimizing the extraction conditions. The best extraction conditions for the simultaneous optimization were an extraction time of 38 min, a solvent concentration of 58%, and a solvent-to-material ratio of 30 mL/g. Under these conditions, the optimized values of yield, TPC, TFC, and DPPH-radical scavenging activity (DPPHIC50) were 18.91%, 154.09 mg GAE/g, 23.76 mg QE/g, and 122.47 μg/mL, respectively. The further HPLC/ESI-MS analysis of the obtained optimized extract revealed the presence of 14 phenolic compounds with piscidic acid, vitexin, and quinic acid as major compounds. These research findings indicate promising applications for efficiently extracting polyphenolic antioxidants, especially in the food industry.


Development of a perfusion process for serum-free adenovirus vector herpes zoster vaccine production.
Herpes zoster is caused by reactivation of the varicella zoster virus (VZV). Researching and developing a herpes zoster vaccine will help to decrease the incidence of herpes zoster. To increase the bioreactor productivity, a serum-free HEK293 cell perfusion process with adenovirus vector herpes zoster (rAd-HZ) vaccine production was developed efficiently using the design of experiment (DoE) method. First, serum-free media for HEK293 cells were screened in both batch and semi-perfusion culture modes. Then, three optimal media were employed in a medium mixture design to improve cell culture performance, and the 1:1 mixture of HEK293 medium and MCD293 medium (named HM293 medium) was identified as the optimal formulation. On the basis of the HM293 medium, the relationship of critical process parameters (CPPs), including the time of infection (TOI), multiplicity of infection (MOI), pH, and critical quality attributes (CQAs) (adenovirus titer (Titer), cell-specific virus yield (CSVY), adenovirus fold expansion (Fold)) of rAd-HZ production was investigated using the DoE approach. Furthermore, the robust setpoint and design space of these CPPs were explored. Finally, the rAd-HZ production process with parameters at a robust setpoint (TOI = 7.2 × 106 cells/mL, MOI = 3.7, and pH = 7.17) was successfully scaled-up to a 3-L bioreactor with an alternating tangential flow system, yielding an adenovirus titer of 3.0 × 1010 IFU/mL, a CSVY of 4167 IFU/cells, a Fold of 1117 at 2 days post infection (dpi). The DoE approach accelerated the development of a HEK293 serum-free medium and of a robust adenovirus production process.


[Intervention for Chest Trauma and Large Vessel Injury].
Trauma is an injury to the body that involves multiple anatomical and pathophysiological changes caused by forces acting from outside the body. The number of patients with trauma is increasing as our society becomes more sophisticated. The importance and demand of traumatology are growing due to the development and spread of treatment and diagnostic technologies. In particular, damage to the large blood vessels of the chest can be life-threatening, and the sequelae are often severe; therefore, diagnostic and therapeutic methods are becoming increasingly important. Trauma to non-aortic vessels of the thorax and aorta results in varying degrees of physical damage depending on the mechanism of the accident and anatomical damage involved. The main damage is hemorrhage from non-aortic vessels of the thorax and aorta, accompanied by hemodynamic instability and coagulation disorders, which can be life-threatening. Immediate diagnosis and rapid therapeutic access can often improve the prognosis. The treatment of trauma can be surgical or interventional, depending on the patient's condition. Among them, interventional procedures are increasingly gaining popularity owing to their convenience, rapidity, and high therapeutic effectiveness, with increasing use in more trauma centers worldwide. Typical interventional procedures for patients with thoracic trauma include embolization for non-aortic injuries and thoracic endovascular aortic repair for aortic injuries. These procedures have many advantages over surgical treatments, such as fewer internal or surgical side effects, and can be performed more quickly than surgical procedures, contributing to improved outcomes for patients with trauma.


Elderly and disabled waiver services: important dimensions of personal care from the client's perspective.
Twenty-five interviews were conducted (18 clients and seven family caregivers) to identify the consumer's most important dimensions of personal care provision. Respondents were generally satisfied with their current aides, but there are unmet needs, including care at additional times of the day or week, and problems, particularly aides arriving late. The most common problems reported from past experience with prior aides included aides who arrived late, did not show up, or who were viewed as lazy. Eight important dimensions were identified, including having aides who are reliable, honest, easy to be around, and who possess a good knowledge base.


Growth rate of human tumour xenografts measured in nude mice by in vivo cast modelling.
Caliper measurement of xenograft tumour volume incurs shape-dependent errors of up to 230 per cent. A more sensitive and accurate method is described for repeated in vivo measurement of tumour volume by cast modelling. The technique can provide electronic data for further analysis and its accuracy and limitations can themselves be determined. Week-on-week changes in tumour volume may be detected with up to 99 per cent confidence.


Scanning electron microscopic analysis of adherent bacterial biofilms associated with peri-implantitis.
Peri-implantitis (PI) is caused by bacteria in the peri-implant space but the consensus on microbial profile is still lacking. Current microbial sampling of PI lesions has largely focused on analyzing bacterial species that have been shed from the implant surface and captured in the pocket fluid. The purpose of the present study was to investigate the morphotypes of bacteria in biofilm covering the implant threads and explore whether certain morphotypes were associated with PI. Fourteen failed implants were removed and instantly processed for scanning electron microscope analysis. The implants were imaged at three equally divided sub-crestal levels of the exposed area. Bacterial morphotypes were identified and quantified by three examiners. Mobility and years in function were correlated to the presence of different morphotypes. The implants demonstrated the presence of variable bacterial morphotypes that did not correlate to disease progression in our study. Some implants were dominated by filaments and others showed the presence of combinations of cocci/rods or spirilles/spirochetes. In general, all implants showed variable morphologic biofilm composition. However, individual implants tended to have similar composition throughout the entire implant. Rods and filaments were dominant morphotypes throughout the surfaces and cocci showed increased presence toward the apical third. There were some differences in the biofilm morphology with mobility and time in function. The profiles of bacterial biofilm morphotypes in failing implants with similar clinical presentations were highly variable. While there were significant differences between implants, similar morphotypes in individual implants were often found throughout the entire surface.


Mycotoxins in cereal grain. Part VIII. Microbiological evaluation of cereal grain quality, connected with mycotoxins occurrence.
Four classes of microbiological quality of cereal grain are proposed. For classification the frequency of kernels contamination with Penicillia, Aspergilli and Phycomycetes is considered as essential. Lots contaminated with mycotoxins (ochratoxins) were usually of IIIrd and IVth class of quality.


Leiomyosarcoma of the kidney. Immunohistological and ultrastructural findings with special emphasis on the growth fraction.
This report deals with a case of renal smooth muscle tumour. Considering the low mitotic rate, this neoplasm would have been classified as leiomyoma. However, it displayed a relatively large growth fraction by immunostaining with the monoclonal antibody Ki-67. This led us to the diagnosis "leiomyosarcoma", which is in line with the unfavourable clinical course. We suggest that assessment of the growth fraction with Ki-67 seems to be more objective than the conventional criteria for determining the malignant potential of smooth muscle tumours.


Properties of base-substituted and carboxyl-esterified analogues of griseolic acid, a potent cAMP phosphodiesterase inhibitor.
Griseolic acid (GA) is a potent cyclic AMP (cAMP) phosphodiesterase (PDE) inhibitor that has an adenine base and two carboxyl groups in its molecule (Nakagawa F, Okazaki T, Naito A, Iijima Y and Yamazaki M, J Antibiot 38: 823-829, 1985). GA analogues were synthesized in which the adenine group was substituted with guanine (6-deamino-2-amino-6-hydroxygriseolic acid, G-GA) or hypoxanthine (6-deamino-6-hydroxygriseolic acid, H-GA). Their inhibitory activities to cyclic GMP (cGMP) PDE and cAMP PDE were compared with GA. For cGMP PDE from rod outer segments of bovine retina, the IC50 values of GA, G-GA and H-GA were 18, 0.040 and 0.12 microM, respectively, with 0.25 microM cGMP as substrate. For type IV PDE isozyme from mouse 3T3 fibroblast cells, the IC50 values of GA, G-GA and H-GA were 0.021, 15 and 11 microM, respectively, with 0.25 microM cAMP as substrate. Thus, GA and G-GA were found to be base-selective inhibitors of type IV PDE of 3T3 cells and type V PDE of bovine retinas, respectively. Esters of carboxylic acids of GA were synthesized in order to increase permeability into cells, and their efficacy was tested by measuring the accumulation of cAMP in 3T3 cells. The dipivaloyloxymethyl ester of GA was found to increase cAMP levels at 0.1 microM, while GA and 3-isobutyl-1-methylxanthine were active only above 100 microM, and the dimethyl ester of GA was inactive. The dipivaloyloxymethyl ester of GA seems to exert its activity after conversion to GA in the cell, since the pivaloyloxymethyl ester was easily hydrolysed by the enzyme action and the dipivaloyloxymethyl ester of GA itself was much less potent an inhibitor of PDE. The dipivaloyloxymethyl ester of GA inhibited thrombin-induced aggregation of platelets and stimulated lipolysis of adipocytes at low concentrations.


[Circadian rhythms in the endocrine system (author's transl)].
In four sections, the contribution reviews data on 24-hour variations of cortisol, growth hormone, prolactine, testosterone, LH and FSH in the human plasma. 1) Experiments on isolated subjects show that the 24-h variation of cortisol is based on an endogenous circadian rhythm which can be independent from the rhythm of sleep and wakefulness. 2) The various pattern each of which has been described in good correspondence by several groups of authors, suggest a series of hormones with a strong circadian component (e.g. cortisol) to those with a weaker (testosterone) or even without a circadian component (FSH). 3) The sleep-wake-cycle influences the phase of hormone rhythms by various degrees; therefore, a uniform reference to sleep time seems preferable to a reference to local time in order to compare several rhythms. 4) The rhythms of cortisol and growth hormone as described in man agree satisfactorily with those in other vertebrate species.


Serum levels of sirtuin 6 are associated with severe community acquired pneumonia in children: An observational study.
The objective of this study was to investigate the role of sirtuin 6 (SIRT6) in severe community acquired pneumonia (CAP) in child patients. This prospective observational research enrolled a total of 75 severe child CAP patients who went to our hospital during April 2016 to December 2020, and 75 mild/moderate CAP child patients were included as control. SIRT6 and inflammatory factors C-reactive protein (CRP), interleukin (IL)-6, and procalcitonin (PCT) were tested by the enzyme linked immunosorbent assay (ELISA). Demographic data including age, sex, as well as clinical symptoms, duration of ICU stay, duration of mechanical ventilation were collected. The routine blood test was conducted for all patients and WBC amount and neutrophil ratio were recorded. The pediatric critical illness score (PCIS) and 1-month mortality were collected. Levels of SIRT6 were remarkably lower in severe CAP patients or deceased patients compared with mild/moderate or survival patients, respectively. Levels of CRP, PCT, and interleukin-6 (IL-6) were markedly higher in severe patients than mild/moderate patients. However, only levels of CRP were significantly higher in deceased CAP patients and serum levels of SIRT6 were negatively correlated with serum levels of CRP, PCT, and IL-6. The higher levels of CRP, PCT and IL-6, as well as higher mortality rate and lower levels of PCIS were found in patients with lower SIRT6 compared with parents with higher SIRT6. SIRT6 had the potential for diagnosis of severe CAP and patients with lower SIRT1 showed shorter 1-month survival. Further, logistic regression showed that only age and CRP were independent risk factors for 1-month mortality of CAP child parents. Down-regulated SIRT6 in severe CAP child patients predicted higher expression of inflammatory factors, severer clinical outcomes and poor prognosis.


A platelet factor that stimulates the proliferation of vascular endothelial cells.
The effects of platelet factors on the growth of cultured porcine aortic endothelial cells were studied. Human platelet lysate stimulated the incorporation of [3H] thymidine into DNA. Gel chromatography on Sephadex G-75 revealed at least two peaks of activity on endothelial cells, the major peak being at an apparent molecular weight of 20,000. This activity was heat-labile and trypsin-sensitive, and did not stimulate the growth of fibro-blasts.


Isotype switching of an immunoglobulin heavy chain transgene occurs by DNA recombination between different chromosomes.
Transgenic mice carrying an immunoglobulin mu heavy chain transgene exhibit isotype switching of the transgene. We have now characterized the mechanism of transgene switching in these mice. The site of mu transgene insertion in one transgenic line has been localized to chromosome 5 using a series of polymorphic endogenous retroviruses as genetic markers in backcross mice. The endogenous immunoglobulin heavy chain locus resides on mouse chromosome 12, which shows that transgene isotype switching can occur between two different chromosomes even though normal antibody gene switching has generally been thought to occur within one chromosome. We find that transgene isotype switching involves interchromosomal DNA recombination, and our data suggest that the same enzymatic mechanisms mediate both normal isotype switch recombination and interchromosomal transgene switching. Our findings also support the notion that the isotype switching mechanism can induce chromosomal translocations such as observed for the c-myc gene in some B cell tumors.


Ferroptosis: An emerging approach for targeting cancer stem cells and drug resistance.
Resistance to chemotherapeutic agents remains a major challenge in the fierce battle against cancer. Cancer stem cells (CSCs) are a small population of cells in tumors that possesses the ability to self-renew, initiate tumors, and cause resistance to conventional anticancer agents. Targeting this population of cells was proven as a promising approach to eliminate cancer recurrence and improve the clinical outcome. CSCs are less susceptible to death by classical anticancer agents inducing apoptosis. CSCs can be eradicated by ferroptosis, which is a non-apoptotic-regulated mechanism of cell death. The induction of ferroptosis is an attractive strategy to eliminate tumors due to its ability to selectively target aggressive CSCs. The current review critically explored the crosstalk and regulatory pathways controlling ferroptosis, which can selectively induce CSCs death. In addition, successful chemotherapeutic agents that achieve better therapeutic outcomes through the induction of ferroptosis in CSCs were discussed to highlight their promising clinical impact.


Geospatial data from a global survey of martian fan-shaped sedimentary landforms.
Data in this article are related to the research article "The global distribution and morphologic characteristics of fan-shaped sedimentary landforms on Mars". We used globally available image and topographic data to document the location of every fan-shaped sedimentary landform on the surface of Mars. We mapped fan outlines and associated drainage basins and collected a number of morphologic metrics. These data can be used as a boundary condition for studies of global scale studies of Mars, including climate and hydrologic modeling. Data files publicly available on Figshare include point shapefile of fan apices, and polygon shapefiles of fan outlines and drainage basins.