Cellular Clearance and Biological Activity of Calciprotein Particles Depend on Their Maturation State and Crystallinity.
Background: The liver-derived plasma protein fetuin-A is a systemic inhibitor of ectopic calcification. Fetuin-A stabilizes saturated mineral solutions by forming colloidal protein-mineral complexes called calciprotein particles (CPP). CPP are initially spherical, amorphous and soft, and are referred to as primary CPP. These particles spontaneously convert into secondary CPP, which are larger, oblongate, more crystalline, and less soluble. CPP mediate excess mineral transport and clearance from circulation. Methods: We studied by intravital two-photon microscopy the clearance of primary vs. secondary CPP by injecting i.v. synthetic fluorescent CPP in mice. We analyzed CPP organ distribution and identified CPP endocytosing cells by immunofluorescence. Cellular clearance was studied using bone marrow-derived mouse wildtype and scavenger receptor A (SRA)-deficient macrophages, as well as human umbilical cord endothelial cells (HUVEC), monocyte-derived macrophages (hMDM), and human aortic endothelial cells (haEC). We employed mouse wildtype and mutant immortalized macrophages to analyze CPP-induced inflammasome activation and cytokine secretion. Results: In live mice, only primary CPP were rapidly cleared by liver sinusoidal endothelial cells (LSEC), whereas primary and secondary CPP were cleared by Kupffer cells. Scavenger receptor A (SRA)-deficient bone marrow macrophages endocytosed secondary CPP less well than did wildtype macrophages. In contrast, primary CPP endocytosis did not depend on the presence of SRA, suggesting involvement of an alternative clearance pathway. CPP triggered TLR4 dependent TNFα and IL-1β secretion in cultured macrophages. Calcium content-matched primary CPP caused twice more IL-1β secretion than did secondary CPP, which was associated with increased calcium-dependent inflammasome activation, suggesting that intracellular CPP dissolution and calcium overload may cause this inflammation. Conclusions: Secondary CPP are endocytosed by macrophages in liver and spleen via SRA. In contrast, our results suggest that primary CPP are cleared by LSEC via an alternative pathway. CPP induced TLR4-dependent TNFα and inflammasome-dependent IL-1β secretion in macrophages suggesting that inflammation and calcification may be considered consequences of prolonged CPP presence and clearance.


Myofibroblast involvement in the allergen-induced late response in mild atopic asthma.
We undertook a detailed cellular and ultrastructural examination of bronchial biopsies from seven allergic asthmatic patients and 10 nonasthmatic control subjects (five atopic and five nonatopic) to determine the nature of the inflammation that occurs during allergen-induced late-phase responses (LPRs). The asthmatic subjects had mild asthma (FEV1 = 94 +/- 9% predicted; mean +/- SEM) and required only intermittent use of beta 2-agonists. Airway mucosal biopsy specimens were obtained on a single occasion from the nonasthmatic controls and on two occasions from the asthmatic subjects, at 24 h after diluent challenge and 24 h after challenge with allergen 3 wk later. The mean maximal decrease in FEV1 during the late response after allergen challenge was 30%, and that after administration of diluent was 4%. In coded plastic sections, subepithelial cells were counted with both light and electron microscopy, and the numbers present were expressed per 0.1 mm2 of tissue. Light microscopy revealed statistically significant increases in the total number of inflammatory cells (P < 0.02) and in "activated fibroblasts" after allergen challenge (P < 0.05). Electron microscopy showed significant increases after allergen challenge in the total number of eosinophils (P < 0.05) and cells with the ultrastructural features of myofibroblasts. The latter cells constituted 1.5% of cells after administration of diluent, and this increased to 15.5% after allergen challenge (P < 0.05). Mast cells were significantly more abundant in the atopic nonasthmatic controls than in the asthmatic subjects after allergen challenge. The study demonstrates that the profile of inflammatory cells in asthma at 24 h after allergen challenge is distinct from that in stable asthma and in nonasthmatic controls, and that migratory cells with a contractile phenotype appear in greater numbers in the late response. We propose that subjects who repeatedly develop a late response have increased numbers of migrating, contractile cells that may contribute to formation of the increased bronchial smooth-muscle mass observed in fatal asthma.


A Review Study on Macrolides Isolated from Cyanobacteria.
Cyanobacteria are rich sources of structurally-diverse molecules with promising pharmacological activities. Marine cyanobacteria have been proven to be true producers of some significant bioactive metabolites from marine invertebrates. Macrolides are a class of bioactive compounds isolated from marine organisms, including marine microorganisms in particular. The structural characteristics of macrolides from cyanobacteria mainly manifest in the diversity of carbon skeletons, complexes of chlorinated thiazole-containing molecules and complex spatial configuration. In the present work, we systematically reviewed the structures and pharmacological activities of macrolides from cyanobacteria. Our data would help establish an effective support system for the discovery and development of cyanobacterium-derived macrolides.


Met-HGF/SF mediates growth arrest and differentiation in T47D breast cancer cells.
Hepatocyte growth factor/scatter factor (HGF/SF) is a pluripotent growth factor that exerts mitogenic, motogenic, and morphogenic effects. To elucidate the cellular mechanisms underlying the pluripotent function of this growth factor, T47D human breast cancer cells were transfected with human hgf/sf. The hgf/sf-positive clones exhibited different levels of biologically functional HGF/SF expression and up-regulation of endogenous Met (HGF/SF receptor) expression. In addition, a constitutive phosphorylation of the receptor on tyrosine residues was detected, establishing a Met-HGF/SF autocrine loop. The autocrine activation of Met caused marked inhibition in cell growth accompanied by cell accumulation at G0/G1. These cells underwent terminal cell differentiation as determined by morphological changes, synthesis of milk proteins such as beta-casein and alpha-lactalbumin, and production of lipid vesicles. Our results demonstrate that Met-HGF/SF, an oncogenic signal transduction pathway, is capable of inducing growth arrest and differentiation in certain breast cancer cells and, thus, may have potential as therapeutic and/or prognostic tools in breast cancer treatment.


Correlation of the long-term neurological outcomes with completeness of surgical evacuation in spontaneous supratentorial intracerebral haemorrhage: a retrospective study.
The treatment of primary spontaneous supratentorial intracerebral haemorrhage (ICH) by evacuation is not supported by randomised controlled trials. We investigate the effectiveness of the completeness of surgical evacuation of spontaneous supratentorial ICH with respect to the functional neurological outcome and mortality. A retrospective review of patients who underwent supratentorial ICH evacuations in the Neurosurgical Unit of the National University Hospital, Singapore, between January 2002 and December 2005 was conducted. Preoperative and postoperative computed tomography images were compared, and the patients or their family members completed follow-up questionnaires two years post surgery, in order to assess the neurological outcome. The patients were subdivided into two groups based on the Glasgow Outcome Scale and haematoma volume. Patients with small pre-evacuation haematoma had a median percentage change in volume and a midline shift of 97.63 percent and 63 percent, respectively. Patients with a large haematoma volume had a median percentage change in volume and midline shift of 99.54 percent and 100 percent, respectively (the p-values for percentage change in volume and midline shift are 0.764 and 0.742, respectively). The median percentage change in volume for the poor outcome subgroup was 97.63 percent, compared to 100 percent for the good outcome subgroup (p-value is 0.288). The median change in midline shift in the poor and good outcome subgroups was 63 percent and 100 percent, respectively (p-value is 0.576). Although not statistically significant with regard to the completeness of haematoma evacuation, a trend toward better outcome with more complete evacuation is observed with ICH.


Vesicular and juxtavesicular serotonin: effects of lysergic acid diethylamide and reserpine.
An extensive analysis of subcellular serotonin (5-HT) compartmentation with and without reserpine was undertaken in order to localize further the effect of LSD on rat brain 5-HT. Modification of the subfractionation procedure resulted in an increase in purity of fractions and a decrease in variability of 5-HT content. With the modified procedure, the administration of LSD produced a significant increase in 5-HT in the nerve-ending fraction prepared from rat whole brain. LSD caused a 50% increase in 5-HT in the vesicular fraction which was recovered after osmotic disruption of nerve-endings. The increase of 5-HT in the vesicular fraction after LSD was not demonstrable in rats treated with reserpine for as long as 2 weeks postreserpine. Instead, with reserpine pretreatment the LSD-induced increase in 5-HT was localized to the intrasynaptosomally derived "end supernatant" as early as 48 hours postreserpine. Thus, an unanticipated "juxtavesicular" site capable of 5-HT retention or binding was detected. In crude subcellular fractions, by contrast, significant increase in 5-HT were not observed with LSD administration until 4 days after reserpine, at which time at least a 50% 5-HT repletion had occurred. This study of drug interactions suggests a juxtavesicular compartment that may be of functional importance in presynaptic binding or transport of 5-HT.


Evidence for transcription attenuation rendering cryptic a sigmaS-dependent promoter of the osmotically regulated proU operon of Salmonella typhimurium.
The osmotically regulated proU locus in Escherichia coli has two promoters, P1 and P2, that are recognized, respectively, by the sigmaS- and sigma70-bearing RNA polymerase holoenzymes. However, the equivalent of the P1 promoter does not appear to exist in Salmonella typhimurium. We demonstrate in this study that wild-type S. typhimurium has a cryptic P1 promoter that is recognized by sigmaS RNA polymerase in vitro and that a 22-bp deletion from +63 to +84 (relative to the start site of transcription) confers sigmaS-dependent in vivo expression of a reporter gene fusion to P1. Primer extension analysis of RNA isolated from cells carrying the wild-type and mutant S. typhimurium proU constructs indicated that a primer which hybridizes proximal to +60 is able to detect P1-initiated transcripts from both constructs but a primer which hybridizes distal to +85 is able to do so only from the latter. Our results suggest that the sigmaS-controlled proU P1 promoter in S. typhimurium may be rendered cryptic because of factor-dependent transcription attenuation within a short distance downstream of the promoter start site.


Myeloid progenitor cluster formation drives emergency and leukaemic myelopoiesis.
Although many aspects of blood production are well understood, the spatial organization of myeloid differentiation in the bone marrow remains unknown. Here we use imaging to track granulocyte/macrophage progenitor (GMP) behaviour in mice during emergency and leukaemic myelopoiesis. In the steady state, we find individual GMPs scattered throughout the bone marrow. During regeneration, we observe expanding GMP patches forming defined GMP clusters, which, in turn, locally differentiate into granulocytes. The timed release of important bone marrow niche signals (SCF, IL-1β, G-CSF, TGFβ and CXCL4) and activation of an inducible Irf8 and β-catenin progenitor self-renewal network control the transient formation of regenerating GMP clusters. In leukaemia, we show that GMP clusters are constantly produced owing to persistent activation of the self-renewal network and a lack of termination cytokines that normally restore haematopoietic stem-cell quiescence. Our results uncover a previously unrecognized dynamic behaviour of GMPs in situ, which tunes emergency myelopoiesis and is hijacked in leukaemia.


Update on surgical treatments for sleep apnea.
Many patients with obstructive sleep apnoea (OSA), as well as the medical community, are seeking alternative therapies to continuous positive airway pressure (CPAP). Where there are problems with CPAP adherence, surgical procedures are an option without the need for adequate compliance. A wide variety of surgical procedures is available, all of which address the differing anatomy and types of patients requiring specific evaluation of the available data. The author performed a literature search up to October 2008, the studies being evaluated according to EBM criteria. The data for some of the methods was limited. Minimally invasive surgery is helpful due to its positive efficacy vs morbidity ratio. While UPPP is still the standard procedure in mild to moderate OSA, its success is difficult to predict and often falls off with the passage of years. Additional upper airway investigations have not yet succeeded in overcoming this disadvantage. Combined surgery of the multi-level is reserved for secondary treatment after CPAP failure. Only tonsillectomy and maxillomandibular advancement are successful enough to be considered a first-line treatment in certain patients.


Novel CYP2D6 and CYP2C19 variants identified in a patient with adverse reactions towards venlafaxine monotherapy and dual therapy with nortriptyline and fluoxetine.
We present a case report of novel variants of CYP2D6 and CYP2C19 identified in a patient who experienced adverse effects during antidepressant therapy. CYP2D6 DNA sequencing revealed that the patient was most likely an intermediate metabolizer, owing to the presence of a novel variant (2579C>T), which gives rise to a premature stop codon in exon 5. Because defects in CYP2C19 may also be important, we sequenced the promoter region and all exons of CYP2C19 and identified a cluster of three novel variants (-13G>A, 7C>T and 10T>C) around exon 1, as well as the more common CYP2C19*2 allele. The presence of multiple genetic lesions in CYP2C19 implies that this patient is potentially a CYP2C19 poor metabolizer, and this was confirmed by haplotype analysis. Combined impairment of CYP2D6 and CYP2C19 activities, we believe, may have contributed to the development of the observed drug responses in the present report.


[The determination of the optimal irradiation dose in the tumor tissue-normal body tissues system].
To estimate the optimal cancerocidal dose, it is necessary to construct the mathematical models describing the probability of radiation sterilization of tumor tissue and that of a radiation-induced complication in the selected system of normal tissues or in the particular normal tissue if its radiosensitivity is higher than that of other tissues. The optimal cancerocidal dose may be estimated while solving a special emergency problem from the proposed criterion by taking into account the limited probabilities of radiation-induced complications in normal organs and tissues if they are damaged.


Cardiotoxicity of fluoropyrimidines in different schedules of administration: a prospective study.
Cardiotoxicity associated with 5-Fluorouracil (5FU) administration has been infrequently reported in literature, albeit various series of acute coronary syndromes have recorded a low but definite incidence of the above toxicity. In the present study, patients undergoing 5FU-based and oral capecitabine (Xeloda-based chemotherapy were tested for the potential development of cardiac-related symptoms during their administration. Six hundred and forty-four patients entered the study. Those experiencing any cardiac-related symptoms during 5FU infusion or oral capecitabine were subjected to ECG and serum cardiac enzymes determination. If cardiotoxicity was confirmed, 5FU infusion or oral capecitabine were interrupted, sublingual nitrates administered and cardiac monitoring initiated, while patients with >two-fold enzyme elevation were followed in a coronary care unit for at least 72 h. Cases with acute myocardial infarction were excluded from further 5FU or oral capecitabine treatment. Overall 26 patients (4.03%) developed symptoms and/or ECG abnormalities due to 5FU and capecitabine. Patients with continuous 5FU infusion presented a higher incidence of cardiotoxicity [14/209; 6.7%, 95% confidence interval (CI) = 3.3-10.1%] than the remaining (7/317; 2.3%, 95% CI = 0.8-3.3%) (P < 0.012). Specifically, an increased incidence of cardiac-related events was encountered in patients with continuous 24-h 5FU + LV infusion for 5 days (12.5%, 95% CI = 2.3-22.7%) rather than in patients with the same schedule without LV (5.3%, 95% CI = 1.95-8.67%) (P < 0.027), as well as in patients with short 5FU + LV administration (2.4%, 95% CI = 0.9-3.9%) (P < 0.019). Overall, 3/54 patients (5.5%, 95% CI = -0.6-11.1%) on oral capecitabine developed cardiac-related events. Seven out of the 20 patients suffered an acute myocardial infarction, 6 developed ischemia only, while 4 more patients had ECG consistent with coronary vasospasm and 3 with conduction disturbances, of which one subsequently died. Patients administered oral capecitabine had a similar incidence of cardiac-related events; 1/22 (4.5%) patients with advanced breast cancer and 2/32 (6.2%) with colorectal cancer. The present study supports the toxic effect of 5-FU on the myocardium, which is largely schedule-dependent, whereas a low but finite risk of such toxicity has been observed with oral capecitabine. A high level of alertness is required when using fluoropyrimidines (i.v. 5FU or oral capecitabine), while their toxic effect on the coronary endothelium and myocardium merits further investigation.


Side effects during long-term treatment with depot antipsychotic medication.
During long-term treatment of schizophrenia with antipsychotic medication, side effects such as weight gain and tardive dyskinesia may develop, while other extrapyramidal side effects may continue. A large multicenter, double-blind, comparative trial of haloperidol decanoate and fluphenazine decanoate has been completed, in which 181 patients took part. The study lasted for 1 year and patients were assessed using the Krawiecka scale, the Montgomery-Asberg scales for schizophrenia and depression, the Simpson-Angus scale for parkinsonism, the AIMS for tardive dyskinesia, and a scale for akathisia. Patients were also weighed at each assessment. The results provide further evidence about the occurrence of weight changes, extrapyramidal side effects, and depression during long-term treatment and about the relative benefits of the two treatments.


Role of diet and food intake in age-related macular degeneration: a systematic review.
A systematic literature review was conducted to evaluate the role of diet and food intake in age-related macular degeneration (AMD). Eighteen high-quality studies were identified. Adherence to a Mediterranean diet had decreased risk of AMD progression. An Oriental diet pattern had decreased association with AMD prevalence, whereas a Western diet pattern had increased association with AMD prevalence. High consumption of vegetables rich in carotenoids and fatty fish containing omega-3 fatty acids was beneficial for those at risk of AMD. Vegetable oils and animal fats containing omega-6 fatty acids, and red/processed meat should be consumed minimally to reduce the risk of AMD progression. High glycaemic index diets and alcohol consumption of greater than two drinks a day had increased association with AMD. As the quality of diet and food intake had a vital role in AMD, the provision of appropriate nutritional advice to those at risk of AMD is recommended.


Value of testicular and sperm profiles in optimizing reproductive success: lessons learned from selective breeding programs of domestic and laboratory animals.
The economic potential of the biotechnology of artificial insemination for genetic improvement and disease control was recognized in the United States 50 years ago. To fully exploit this technology in dairy cattle, researchers worked closely with industry to study testicular function, best methods for harvesting and evaluating semen, techniques for preserving and inseminating sperm and finally, methods for mass recording of fertility information. Methods of testicular and semen evaluation have been developed which are highly correlated with other reproductive characteristics, including fertility. This has resulted in an unprecedented opportunity to study experimentally imposed conditions and naturally occurring environmental conditions upon reproductive efficiency in the male. The selection and testing of sires has resulted practically in the elimination of genes which produce lethal or severely depressed performance. The use of one male for the controlled insemination of many females also has facilitated studies on reproductive problems in the female. While less selective breeding programs have been conducted with the rabbit relative to reproductive function, the rabbit has been used extensively as a model for large domestic animals and humans. The rabbit is the smallest laboratory animal which can be trained easily to ejaculate into an artificial vagina and use the semen for insemination. This permits longitudinal and correlative studies to be performed relating testis function and epididymal transport to sperm output, semen quality and fertility. Spermatogenesis of both bulls and rabbits is sensitive to toxicants. The effects can be monitored by testicular biopsy and cytological evaluation of the testis, by semen collection and semen evaluation, and by in vitro and in vivo fertility testing. Gross effects on the testis are detectable in the live animal by measuring testis size and by ultrasound profiles.


Seizure Prediction Using Undulated Global and Local Features.
In this study, a seizure prediction method is proposed based on a patient-specific approach by extracting undulated global and local features of preictal/ictal and interictal periods of EEG signals. The proposed method consists of feature extraction, classification, and regularization. The undulated global feature is extracted using phase correlation between two consecutive epochs of EEG signals and an undulated local feature is extracted using the fluctuation and deviation of EEG signals within the epoch. These features are further used for classification of preictal/ictal and interictal EEG signals. A regularization technique is applied on the classified outputs for the reduction of false alarms and improvement of the overall prediction accuracy (PA). The experimental results confirm that the proposed method provides high PA (i.e., 95.4%) with low false positive per hour using intracranial EEG signals in different brain locations of 21 patients from a benchmark dataset. Combining global and local features enables the transition point to be determined between different types of signals with greater accuracy, resulting successful versus unsuccessful prediction of seizure. The theoretical contribution of this study may provide an opportunity for the development of a clinical device to predict forthcoming seizure in real time.


International prospective cohort study of microvascular angina - Rationale and design.
Patients with signs and symptoms of myocardial ischemia and non-obstructive coronary artery disease (CAD) frequently have coronary functional abnormalities, including coronary microvascular dysfunction. Those with the latter are grouped under the term "microvascular angina" (MVA). Although diagnostic criteria exist for MVA, as recently proposed by our COVADIS (COronary VAsomotor Disorders International Study) group and the condition has been increasingly recognized in clinical practice, the clinical characteristics and long-term prognosis of MVA patients in the current era remain to be fully elucidated. In the present study, we aimed to prospectively assess the clinical characteristics and long-term prognosis of MVA subjects in the current era in an international, multicenter, observational, and prospective registry study. A total of 15 medical centers across 7 countries (USA, UK, Germany, Spain, Italy, Australia, and Japan) enrolled subjects fulfilling the COVADIS diagnostic criteria for MVA as follows; (1) signs and/or symptoms of myocardial ischemia, (2) absence of obstructive CAD, and (3) objective evidence of myocardial ischemia and/or coronary microvascular dysfunction. The primary endpoint was the composite of major cardiovascular events (MACE), including cardiovascular death, non-fatal myocardial infarction, non-fatal stroke, hospitalization due to heart failure or unstable angina. Between July 2015 and December 2018, a total of 706 subjects with MVA (M/F 256/450, 61.1 ± 11.8 [SD] yrs.) were registered. Subjects will be followed for at least 1 year. The present study will provide important information regarding the clinical characteristics, management, and long-term prognosis of MVA patients in the current era.


A comparison of inflammatory and oxidative stress markers in adipose tissue from weight-matched obese male and female mice.
Expansion of intra-abdominal adipose tissue and the accompanying inflammatory response has been put forward as a unifying link between obesity and the development of chronic diseases. However, an apparent sexual dimorphism exists between obesity and chronic disease risk due to differences in the distribution and abundance of adipose tissue. A range of experimental protocols have been employed to demonstrate the role of estrogen in regulating health benefits; however, most studies are confounded by significant differences in body weight and adiposity. Therefore, the purpose of this study was to compare weight-matched obese male and female mice to determine if the sex-dependent health benefits remain when body weight is similar. The development of obesity in female mice receiving a high-fat diet was delayed; however, subsequent comparisons of weight-matched obese mice revealed greater adiposity in obese female mice. Despite excess adiposity and enlarged adipocyte size, obese females remained more glucose tolerant than weight-matched male mice, and this benefit was associated with increased expression of adiponectin and reductions in immune cell infiltration and oxidative stress in adipose tissue. Therefore, the protective benefits of estrogen persist in the obese state and appear to improve the metabolic phenotype of adipose tissue and the individual.


Decline in coccolithophore diversity and impact on coccolith morphogenesis along a natural CO2 gradient.
A natural pH gradient caused by marine CO2 seeps off Vulcano Island (Italy) was used to assess the effects of ocean acidification on coccolithophores, which are abundant planktonic unicellular calcifiers. Such seeps are used as natural laboratories to study the effects of ocean acidification on marine ecosystems, since they cause long-term changes in seawater carbonate chemistry and pH, exposing the organisms to elevated CO2 concentrations and therefore mimicking future scenarios. Previous work at CO2 seeps has focused exclusively on benthic organisms. Here we show progressive depletion of 27 coccolithophore species, in terms of cell concentrations and diversity, along a calcite saturation gradient from Ωcalcite 6.4 to <1. Water collected close to the main CO2 seeps had the highest concentrations of malformed Emiliania huxleyi. These observations add to a growing body of evidence that ocean acidification may benefit some algae but will likely cause marine biodiversity loss, especially by impacting calcifying species, which are affected as carbonate saturation falls.


Octopamine mimics the effects of parasitism on the foregut of the tobacco hornworm Manduca sexta.
The parasitic braconid wasp Cotesia congregata lays its eggs inside the body of the larval stage of its host, the moth Manduca sexta. The Cotesia congregata larvae develop within the hemocoel of their host until their third instar, when they emerge and spin cocoons and pupate on the outer surface of the caterpillar. From this time until their death approximately 2 weeks later, the Manduca sexta larvae show striking behavioral changes that include dramatic declines in spontaneous activity and in the time spent feeding. Coincident with these behavioral changes, it is known that octopamine titers in the hemolymph of the host become elevated by approximately 6.5-fold. Octopamine is an important modulator of neural function and behavior in insects, so we examined hosts for neural correlates to the behavioral changes that occur at parasite emergence. We found that, in addition to the changes reported earlier, after parasite emergence (post-emergence), Manduca sexta larvae also showed marked deficits in their ability to ingest food because of a disruption in the function of the frontal ganglion that results in a significant slowing or the absence of peristaltic activity in the foregut. This effect could be produced in unparasitized fifth-instar larvae by application of blood from post-emergence parasitized larvae or of 10(-6)mol l(-1)d,l-octopamine (approximately the level in the hemolymph of post-emergence larvae). In contrast, blood from parasitized larvae before their parasites emerge or from unparasitized fifth-instar larvae typically had no effect on foregut activity. The effects of either post-emergence parasitized blood or 10(-6)mol l(-1) octopamine could be blocked by the octopamine antagonists phentolamine (at 10(-5)mol l(-1)) or mianserin (at 10(-7)mol l(-1)).


Oxidation of gasoline oxygenates by closely related non-haem-iron alkane hydroxylases in Pseudomonas mendocina KR1 and other n-octane-utilizing Pseudomonas strains.
Pseudomonas mendocina KR1 oxidizes the gasoline oxygenate methyl tertiary butyl ether (MTBE) to tertiary butyl alcohol (TBA) during growth on C5 -C8 n-alkanes. We have further explored oxidation of ether oxygenates by this strain to help identify the enzyme that catalyses these reactions. High levels of MTBE-oxidizing activity occurred in resting cells grown on C5 -C8 n-alkanes. Lower activities occurred in cells grown on longer-chain n-alkanes (C9 -C11 ) and 1°-alcohols (C5 -C10 ). N-octane-grown cells also oxidized tertiary amyl methyl ether (TAME) to tertiary amyl alcohol (TAA), but did not oxidize ethyl tertiary butyl ether (ETBE), TBA or TAA. A 39 kDa polypeptide in whole cell extracts of n-octane-grown cells strongly cross-reacted with an anti-AlkB polyclonal antiserum in an SDS-PAGE/immunoblot. This polypeptide was absent or less abundant in cells grown on dextrose, dextrose plus dicyclopropylketone or 1-octanol. N-octane-grown cells of Pseudomonas aeruginosa strains KSLA-473 and ATCC 17423 oxidized MTBE and TAME but not ETBE. N-hexadecane-grown cells of these strains and strain PAO1 did not oxidize any of the oxygenates tested. Our results indicate ether oxygenate-degrading activity in alkane-utilizing pseudomonads is consistently observed with close homologues of the GPo1 non-haem-iron alkane hydroxylases but is otherwise not a consistent catalytic feature of these diverse enzymes.


Behavioral and emotional adjustment, family functioning, academic performance, and social relationships in children with selective mutism.
This study addressed four questions which parents of children with selective mutism (SM) frequently ask: (1) Is SM associated with anxiety or oppositional behavior? (2) Is SM associated with parenting and family dysfunction? (3) Will my child fail at school? and (4) Will my child make friends or be teased and bullied? In comparison to a sample of 52 community controls, 52 children with SM were more anxious, obsessive, and prone to somatic complaints. In contrast, children with SM were less oppositional and evidenced fewer attentional difficulties at school. We found no group differences in family structure, economic resources, family functioning, maternal mood difficulties, recreational activities, or social networks. While parents reported no differences in parenting strategies, children with SM were described as less cooperative in disciplinary situations. The academic (e.g., reading and math) and classroom cooperative skills of children with SM did not differ from controls. Parents and teachers reported that children with SM had significant deficits in social skills. Though teachers and parents rated children with SM as less socially assertive, neither teachers nor parents reported that children with SM were victimized more frequently by peers.


Cell density-dependent modulation of basic fibroblast growth factor expression by human interferon-beta.
The production of basic fibroblast growth factor (bFGF) by human renal cell carcinoma (HRCC) is associated with induction of angiogenesis. Incubation of HRCC cells with human interferon alpha (IFN-alpha) or interferon beta (IFN-beta) downregulates the expression of bFGF and, hence, angiogenesis. The purpose of this study was to analyze the downregulation of the expression of bFGF in HRCC cells by IFN-alpha and IFN-beta. Human HRCC SN12PM6 cells cultured under sparse conditions expressed 3-7-fold higher levels of steady-state bFGF-specific mRNA transcripts and cellular bFGF protein than did confluent cultures. IFN-alpha or IFN-beta inhibited the steady-state expression of bFGF mRNA transcripts and cellular bFGF protein in a concentration-dependent manner in sparse but not confluent cultures. Moreover, IFN-beta downregulated the transcription rate of bFGF genes and inhibited the de novo synthesis of bFGF protein only in sparse cultures. The results demonstrate that the inhibitory effects of IFN- and -beta on bFGF expression are cell-density dependent.


Excimer Laser Sheath-Assisted Retrieval of "Closed-Cell" Design Inferior Vena Cava Filters.
Background Numerous reports have shown that inferior vena cava filters are associated with clinically significant adverse events. Complicating factors, such as caval incorporation, may lead to technical challenges at retrieval. The use of advanced techniques including the laser sheath have increased technical success rates; however, the data are limited on which filter types necessitate and benefit from its use. Methods and Results From October 2011 to September 2019, patients with inferior vena cava filter dwell times >6 months or with prior failed retrievals were considered for laser sheath-assisted retrieval. Standard and nonlaser advanced retrieval techniques were attempted first; if the filter could not be safely or successfully detached from the caval wall using these techniques, the laser sheath was used. Technical success, filter type, necessity for laser sheath application based on "open" versus "closed-cell" filter design, dwell times, and adverse events were evaluated. A total of 441 patients (216 men; mean age, 54 years) were encountered. Mean dwell times for all filters was 56.6 months, 54.4 among closed-cell filters and 58.5 among open-cell filters (P=0.63). Technical success of retrieval was 98%, with the laser sheath required in 143 cases (40%). Successful retrieval of closed-cell filters required laser sheath assistance in 60% of cases as compared with 7% of open-cell filters (odds ratio, 20.1; P<0.01). In closed-cell inferior vena cava filters, dwell time was significantly associated with need for laser, requiring it in 64% of retrievals with dwell times >6 months (P=0.01). One major adverse event occurred among laser sheath retrievals when a patient required a 2-day inpatient admission for a femoral access site hemorrhage. Conclusions Closed-cell filters may necessitate the use of the laser sheath for higher rates of successful and safe retrieval.


Gliding motility in bacteria: insights from studies of Myxococcus xanthus.
Gliding motility is observed in a large variety of phylogenetically unrelated bacteria. Gliding provides a means for microbes to travel in environments with a low water content, such as might be found in biofilms, microbial mats, and soil. Gliding is defined as the movement of a cell on a surface in the direction of the long axis of the cell. Because this definition is operational and not mechanistic, the underlying molecular motor(s) may be quite different in diverse microbes. In fact, studies on the gliding bacterium Myxococcus xanthus suggest that two independent gliding machineries, encoded by two multigene systems, operate in this microorganism. One machinery, which allows individual cells to glide on a surface, independent of whether the cells are moving alone or in groups, requires the function of the genes of the A-motility system. More than 37 A-motility genes are known to be required for this form of movement. Depending on an additional phenotype, these genes are divided into two subclasses, the agl and cgl genes. Videomicroscopic studies on gliding movement, as well as ultrastructural observations of two myxobacteria, suggest that the A-system motor may consist of multiple single motor elements that are arrayed along the entire cell body. Each motor element is proposed to be localized to the periplasmic space and to be anchored to the peptidoglycan layer. The force to glide which may be generated here is coupled to adhesion sites that move freely in the outer membrane. These adhesion sites provide a specific contact with the substratum. Based on single-cell observations, similar models have been proposed to operate in the unrelated gliding bacteria Flavobacterium johnsoniae (formerly Cytophaga johnsonae), Cytophaga strain U67, and Flexibacter polymorphus (a filamentous glider). Although this model has not been verified experimentally, M. xanthus seems to be the ideal organism with which to test it, given the genetic tools available. The second gliding motor in M. xanthus controls cell movement in groups (S-motility system). It is dependent on functional type IV pili and is operative only when cells are in close proximity to each other. Type IV pili are known to be involved in another mode of bacterial surface translocation, called twitching motility. S-motility may well represent a variation of twitching motility in M. xanthus. However, twitching differs from gliding since it involves cell movements that are jerky and abrupt and that lack the organization and smoothness observed in gliding. Components of this motor are encoded by genes of the S-system, which appear to be homologs of genes involved in the biosynthesis, assembly, and function of type IV pili in Pseudomonas aeruginosa and Neisseria gonorrhoeae. How type IV pili generate force in S-motility is currently unknown, but it is to be expected that ongoing physiological, genetic, and biochemical studies in M. xanthus, in conjunction with studies on twitching in P. aeruginosa and N. gonorrhoeae, will provide important insights into this microbial motor. The two motility systems of M. xanthus are affected to different degrees by the MglA protein, which shows similarity to a small GTPase. Bacterial chemotaxis-like sensory transduction systems control gliding motility in M. xanthus. The frz genes appear to regulate gliding movement of individual cells and movement by the S-motility system, suggesting that the two motors found in this bacterium can be regulated to result in coordinated multicellular movements. In contrast, the dif genes affect only S-system-dependent swarming.


Cold Acclimation and Deacclimation of Two Garden Rose Cultivars Under Controlled Daylength and Temperature.
Low temperature stress is an important abiotic stress for garden roses in northern regions. Two garden rose cultivars ('Dagmar Hastrup' and 'Chandos Beauty') were selected to study the role of dehydrin and of carbohydrate metabolism during cold acclimation and deacclimation under the controlled daylength and temperature. The presence of bud dormancy was also observed as this could prevent budburst during warm spells. Both cultivars showed a similar changing pattern of cold acclimation and deacclimation and did not differ in their lowest LT50 values. Dehydrin (RhDHN5) was up-regulated by low temperatures and not by dehydration stress as the stem water content remained stable during the treatments. Total soluble sugars accumulated with a transient up-regulation of RhBAM3 (a key gene in starch hydrolysis) for 'Dagmar Hastrup' at 2°C and a strong expression under both 2 and -3°C for 'Chandos Beauty'. At 2 and -3°C, raffinose and stachyose strongly accumulated though the up-regulation of RhRS6 and RhGK differed in the cultivars. Although similar cold hardiness levels were reached, carbohydrate metabolism in response to cold stress is different in the two cultivars. Increasing the temperature after a cold period resulted in fast deacclimation as found by the downregulation of RhDHN5 and RhBAM3, the decrease of raffinose and stachyose. Bud endodormancy was hardly present in both cultivars.


HPV16 Load Is a Potential Biomarker to Predict Risk of High-Grade Cervical Lesions in High-Risk HPV-Infected Women: A Large Longitudinal French Hospital-Based Cohort Study.
High-risk HPV (hrHPV) testing has been implemented as a primary screening tool for cervical cancer in numerous countries. However, there is still a need for relevant triage strategies to manage hrHPV positive women to avoid excessive referral to colposcopy. The objective of this study was to assess, in women infected by hrHPV and presenting no or mild cytological abnormalities, HPV16 and HPV18 viral loads to predict the development of cervical high-grade lesion. Among 2102 women positive for hrHPV, 885 had no lesion or mild cytological abnormalities at baseline and had at least one follow-up (FU) visit. HPV16 and HPV18 prevalence was 25.9% and 8.4%, respectively. Of those women, 15% developed a high-grade lesion during the FU. An HPV16 viral load cut-off set at 3.2 log10GE/103 cells permitted to identify a subgroup of women at high risk of developing high-grade cervical lesion (HR = 2.67; 95% CI 1.80-3.97; p ≤ 0.0001). No specific HPV18 viral load threshold could have been defined in regard to the present study. In multivariate analysis, HPV16 load (absence/log10GE/103 cells < 3.2 vs. ≥3.2), RLU/PC 239 (1-100 pg/mL vs. >100 pg/mL) and cytology (normal vs abnormal) were independently associated with a significant increased risk of high-grade lesion development and were used to construct the prognostic score. In conclusion, HPV16 load is a relevant biomarker to identify women at high risk for developing cervical precancerous lesions.


Development of a VR Application for Binge Eating Treatment: Identification of Contexts and Cues Related to Bingeing Behavior in Spanish Italian Patients.
The objective of this study was to identify frequent situations and specific cues that produce the craving to binge in Spanish and Italian samples of patients with eating disorders (ED). There were two main aims: to assess transcultural differences in the contexts and cues that elicit food craving; and to develop valid, reliable VR environments for effective cue-exposure therapy (CET) for patients from both countries. Twenty-six Spanish and 75 Italian ED patients completed an ad hoc questionnaire to assess contexts and cues that trigger the craving to binge. No differences between groups were found. All patients reported experiencing higher levels of craving in the afternoon/early evening and in the late evening/night, between meals, when alone, and more frequently at the end of the week. Being in the dining room, the kitchen, the bedroom, the bakery and the supermarket were the specific situations that produced the highest levels of craving to binge. We used the questionnaire results to develop a virtual reality application for CET.


Musical Instrument Practice Predicts White Matter Microstructure and Cognitive Abilities in Childhood.
Musical training has been associated with advantages in cognitive measures of IQ and verbal ability, as well as neural measures including white matter microstructural properties in the corpus callosum (CC) and the superior longitudinal fasciculus (SLF). We hypothesized that children who have musical training will have different microstructural properties in the SLF and CC. One hundred children aged 7.9-9.9 years (mean age 8.7) were surveyed for their musical activities, completed neuropsychological testing for general cognitive abilities, and underwent diffusion tensor imaging (DTI) as part of a larger study. Children who play a musical instrument for more than 0.5 h per week (n = 34) had higher scores on verbal ability and intellectual ability (standardized scores from the Woodcock-Johnson Tests of Cognitive Abilities), as well as higher axial diffusivity (AD) in the left SLF than those who did not play a musical instrument (n = 66). Furthermore, the intensity of musical practice, quantified as the number of hours of music practice per week, was correlated with axial diffusivity (AD) in the left SLF. Results are not explained by age, sex, socio-economic status, or physical fitness of the participants. The results suggest that the relationship between musical practice and intellectual ability is related to the maturation of white matter pathways in the auditory-motor system. The findings suggest that musical training may be a means of improving cognitive and brain health during development.


ProtoSteer: Steering Deep Sequence Model with Prototypes.
Recently we have witnessed growing adoption of deep sequence models (e.g. LSTMs) in many application domains, including predictive health care, natural language processing, and log analysis. However, the intricate working mechanism of these models confines their accessibility to the domain experts. Their black-box nature also makes it a challenging task to incorporate domain-specific knowledge of the experts into the model. In ProtoSteer (Prototype Steering), we tackle the challenge of directly involving the domain experts to steer a deep sequence model without relying on model developers as intermediaries. Our approach originates in case-based reasoning, which imitates the common human problem-solving process of consulting past experiences to solve new problems. We utilize ProSeNet (Prototype Sequence Network), which learns a small set of exemplar cases (i.e., prototypes) from historical data. In ProtoSteer they serve both as an efficient visual summary of the original data and explanations of model decisions. With ProtoSteer the domain experts can inspect, critique, and revise the prototypes interactively. The system then incorporates user-specified prototypes and incrementally updates the model. We conduct extensive case studies and expert interviews in application domains including sentiment analysis on texts and predictive diagnostics based on vehicle fault logs. The results demonstrate that involvements of domain users can help obtain more interpretable models with concise prototypes while retaining similar accuracy.


[Necessary changes for advancing nursing as caring science].
The article aimed to reflect upon the challenges involved in strengthening Nursing as a caring science. It is founded on the sociological theory, connecting three approaches: the historical-dialectic materialism perspective about the working process in health care and nursing; the sociology of professions from a critical perspective; and the philosophy of science. The discussion is organized considering the aspects of Nursing as a discipline, work and health care profession. It sustains that knowledge production should be driven both by the purpose of Nursing work which is providing care to human beings with health needs and to advocate for the indispensable work conditions to a safe and responsible practice. It concludes that to strengthening Nursing it is necessary to produce knowledge to support nursing care and the political actions defending safe work conditions, the universal right to health as well safe and high quality care.


Investigation of the ¹H-NMR based urine metabolomic profiles of IUGR, LGA and AGA newborns on the first day of life.
(1)H-NMR spectroscopy coupled with multivariate statistical analysis was used for the first time to compare the urinary NMR metabolic profiles of neonates with intrauterine growth retardation (IUGR) and large for gestational age (LGA). For the sake of comparison, infants who were adequate for gestational age (AGA) were also analyzed. Pattern recognition methods, including Principal Component Analyses (PCA), Partial Least Squares Discriminant Analysis (PLS-DA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA), were used to analyze NMR data. Clear differences among the metabolic profiles of AGA, IUGR and LGA were observed. The main metabolites responsible for these differentiations were identified as myo-inositol, creatinine, creatine, citrate, urea and glycine. In particular, among these, myo-inositol may be a potential biomarker of an altered glucose metabolism during fetal development both in IUGR and LGA. This study highlights the applicability of NMR-based metabolomics for improving the understanding of the relations among nutrition, integrated metabolism and health in neonatology.


[Intrauterine insemination versus programmed intercourse in cycles of controlled ovaric hyperstimulation].
Assisted reproduction techniques are used more and more frequently in the treatment of coulples with infertility diagnosis. To analyze the intrauterine insemination (IUI) value in controlled ovarian hyperstimulation cycles (COH). An analytic, comparative, retrospective and longitudinal case-control study was performed. COH with IUI (group I) or with timed intercourse (TI) (group II) cycles from January 1st 2004 to December 31st 2006 were analyzed. Infertile patients aged between 24 and 42 years (group I) and between 23 and 36 years (group II) were included. The following variables were analyzed: Age, type, etiology and duration of infertility, sperm density and motility after capacitation or seminal analysis, number of total and mature follicles, endometrial thickness, gonadotropin type and dosage, insemination or TI cycle day. Pregnant vs non pregnant and pregnancies in both groups were analyzed. Results were analyzed with the STATA 7.0 and SPSS 12.0 programs. 873 COH+I UI cycles in 539 couples and 246 COH+ TI in 138 patients were analyzed the pregnancy rates per cycle were 13.1% and 5.2% for each group respectively. The mean woman's age was of 32.9 +/- 3.5 and 31.8 +/- 2.7 years for groups I and II respectively. There were significant differences in sperm density in both groups. The remaining variables showed no statistical differences. IUI versus TI do not seems to be superior with respect to the pregnancy rates in COH cycles.


Serum gamma-glutamyltransferase as a prognostic indicator of neonatal viability in nondomestic ruminants.
Rapid assessment of immune status in neonatal ruminants of endangered species facilitates early intervention in cases of inadequate passive transfer of maternal immunoglobulins. Serum gamma-glutamyltransferase (GGT) was used to evaluate suspected passive transfer status in 25 North Indian muntjac (Muntiacus muntjak vaginalis), 45 Cretan goats (Capra algagrus cretica), 20 white-lipped deer (Cervus albirostris), 25 Mhorr gazelles (Gazella dama mhorr), and 31 Soemmerring's gazelles (Gazella soemmerringi soemmerringi). Serum GGT, measured within 48 hr of birth, was compared with clinical condition at 5 days of age. Neonatal Soemmerring's and Mhorr gazelles with GGT > 600 U/L were likely to survive without medical intervention, whereas GGT < 400 U/L was a good indicator that the gazelle neonate would need medical intervention. Neonatal muntjac with GGT > 200 U/L were also likely to survive without medical intervention. Because there is no gold standard for evaluating passive transfer status in neonatal nondomestic ruminants, it is recommended to evaluate the results of more than one diagnostic test, as well as clinical condition, in considering health status and disposition of neonatal ruminants of endangered species.


AI software detection of large vessel occlusion stroke on CT angiography: a real-world prospective diagnostic test accuracy study.
Artificial intelligence (AI) software is increasingly applied in stroke diagnostics. However, the actual performance of AI tools for identifying large vessel occlusion (LVO) stroke in real time in a real-world setting has not been fully studied. To determine the accuracy of AI software in a real-world, three-tiered multihospital stroke network. All consecutive head and neck CT angiography (CTA) scans performed during stroke codes and run through an AI software engine (Viz LVO) between May 2019 and October 2020 were prospectively collected. CTA readings by radiologists served as the clinical reference standard test and Viz LVO output served as the index test. Accuracy metrics were calculated. Of a total of 1822 CTAs performed, 190 occlusions were identified; 142 of which were internal carotid artery terminus (ICA-T), middle cerebral artery M1, or M2 locations. Accuracy metrics were analyzed for two different groups: ICA-T and M1 ±M2. For the ICA-T/M1 versus the ICA-T/M1/M2 group, sensitivity was 93.8% vs 74.6%, specificity was 91.1% vs 91.1%, negative predictive value was 99.7% vs 97.6%, accuracy was 91.2% vs 89.8%, and area under the curve was 0.95 vs 0.86, respectively. Detection rates for ICA-T, M1, and M2 occlusions were 100%, 93%, and 49%, respectively. As expected, the algorithm offered better detection rates for proximal occlusions than for mid/distal M2 occlusions (58% vs 28%, p=0.03). These accuracy metrics support Viz LVO as a useful adjunct tool in stroke diagnostics. Fast and accurate diagnosis with high negative predictive value mitigates missing potentially salvageable patients.


Age-Related Variations in Regional White Matter Volumetry and Microstructure During the Post-adolescence Period: A Cross-Sectional Study of a Cohort of 1,713 University Students.
Human brain white matter undergoes a protracted maturation that continues well into adulthood. Recent advances in diffusion-weighted imaging (DWI) methods allow detailed characterizations of the microstructural architecture of white matter, and they are increasingly utilized to study white matter changes during development and aging. However, relatively little is known about the late maturational changes in the microstructural architecture of white matter during post-adolescence. Here we report on regional changes in white matter volume and microstructure in young adults undergoing university-level education. As part of the MRi-Share multi-modal brain MRI database, multi-shell, high angular resolution DWI data were acquired in a unique sample of 1,713 university students aged 18-26. We assessed the age and sex dependence of diffusion metrics derived from diffusion tensor imaging (DTI) and neurite orientation dispersion and density imaging (NODDI) in the white matter regions as defined in the John Hopkins University (JHU) white matter labels atlas. We demonstrate that while regional white matter volume is relatively stable over the age range of our sample, the white matter microstructural properties show clear age-related variations. Globally, it is characterized by a robust increase in neurite density index (NDI), and to a lesser extent, orientation dispersion index (ODI). These changes are accompanied by a decrease in diffusivity. In contrast, there is minimal age-related variation in fractional anisotropy. There are regional variations in these microstructural changes: some tracts, most notably cingulum bundles, show a strong age-related increase in NDI coupled with decreases in radial and mean diffusivity, while others, mainly cortico-spinal projection tracts, primarily show an ODI increase and axial diffusivity decrease. These age-related variations are not different between males and females, but males show higher NDI and ODI and lower diffusivity than females across many tracts. These findings emphasize the complexity of changes in white matter structure occurring in this critical period of late maturation in early adulthood.


Dipeptidylpeptidase-4 inhibitors (gliptins): focus on drug-drug interactions.
Patients with type 2 diabetes mellitus (T2DM) are generally treated with many pharmacological compounds and are exposed to a high risk of drug-drug interactions. Indeed, blood glucose control usually requires a combination of various glucose-lowering agents, and the recommended global approach to reduce overall cardiovascular risk generally implies administration of several protective compounds, including HMG-CoA reductase inhibitors (statins), antihypertensive compounds and antiplatelet agents. New compounds have been developed to improve glucose-induced beta-cell secretion and glucose control, without inducing hypoglycaemia or weight gain, in patients with T2DM. Dipeptidylpeptidase-4 (DPP-4) inhibitors are novel oral glucose-lowering agents, which may be used as monotherapy or in combination with other antidiabetic compounds, metformin, thiazolidinediones or even sulfonylureas. Sitagliptin, vildagliptin and saxagliptin are already on the market, either as single agents or in fixed-dose combined formulations with metformin. Other compounds, such as alogliptin and linagliptin, are in a late phase of development. This review summarizes the available data on drug-drug interactions reported in the literature for these five DDP-4 inhibitors: sitagliptin, vildagliptin, saxagliptin, alogliptin and linagliptin. Possible pharmacokinetic interferences have been investigated between each of these compounds and various pharmacological agents, which were selected because there are other glucose-lowering agents (metformin, glibenclamide [glyburide], pioglitazone/rosiglitazone) that may be prescribed in combination with DPP-4 inhibitors, other drugs that are currently used in patients with T2DM (statins, antihypertensive agents), compounds that are known to interfere with the cytochrome P450 (CYP) system (ketoconazole, diltiazem, rifampicin [rifampin]) or with P-glycoprotein transport (ciclosporin), or agents with a narrow therapeutic safety window (warfarin, digoxin). Generally speaking, almost no drug-drug interactions or only minor drug-drug interactions have been reported between DPP-4 inhibitors and any of these drugs. The gliptins do not significantly modify the pharmacokinetic profile and exposure of the other tested drugs, and the other drugs do not significantly alter the pharmacokinetic profile of the gliptins or exposure to these. The only exception concerns saxagliptin, which is metabolized to an active metabolite by CYP3A4/5. Therefore, exposure to saxagliptin and its primary metabolite may be significantly modified when saxagliptin is coadministered with specific strong inhibitors (ketoconazole, diltiazem) or inducers (rifampicin) of CYP3A4/5 isoforms. The absence of significant drug-drug interactions could be explained by the favourable pharmacokinetic characteristics of DPP-4 inhibitors, which are not inducers or inhibitors of CYP isoforms and are not bound to plasma proteins to a great extent. Therefore, according to these pharmacokinetic findings, which were generally obtained in healthy young male subjects, no dosage adjustment is recommended when gliptins are combined with other pharmacological agents in patients with T2DM, with the exception of a reduction in the daily dosage of saxagliptin when this drug is used in association with a strong inhibitor of CYP3A4/A5. It is worth noting, however, that a reduction in the dose of sulfonylureas is usually recommended when a DPP-4 inhibitor is added, because of a pharmacodynamic interaction (rather than a pharmacokinetic interaction) between the sulfonylurea and the DPP-4 inhibitor, which may result in a higher risk of hypoglycaemia. Otherwise, any gliptin may be combined with metformin or a thiazolidinedione (pioglitazone, rosiglitazone), leading to a significant improvement in glycaemic control without an increased risk of hypoglycaemia or any other adverse event in patients with T2DM. Finally, the absence of drug-drug interactions in clinical trials in healthy subjects requires further evidence from large-scale studies, including typical subjects with T2DM - in particular, multimorbid and geriatric patients receiving polypharmacy.


Entrapment of andrographolide in cross-linked alginate pellets: II. Physicochemical characterization to study the pelletization of andrographolide.
This paper deals with the characterization of pellets containing andrographolide in two parts. The first part deals with characterization of the pellets to ascertain the identity and integrity of andrographolide. Part two involves characterization of the pellets containing Andrographis paniculata extract (33.3%) prepared in the paper I for their micromeritic properties like Particle size, Particle size distribution, Sphericity measurements like Shape ratio and Aspect ratio, Tapped density, Compressibility index, Hausner's ratio and Angle of repose. In addition, our aim was also to derive information about the mechanism of gelation with entrapment of andrographolide to supplement results obtained about the release mechanisms deduced in paper I. Since this work requires use of techniques like FTIR, FTRaman, MTDSC and XRPD, it was necessary to prepare alginate micropellets using pure andrographolide (99.89%) rather than the multicomponent extract using the same procedure discussed in paper I. The integrity of the drug was maintained in the cross-linked micropellets as was seen in the MTDSC and FTIR spectra supported by the FTRaman spectra. The depolymerisation transitions, the reversing and non-reversing heat flow signals were determined using the MTDSC and interpreted to study the mechanism of pelletization. The MTDSC profiles also confirmed the integrity of the drug by exhibiting a sharp endotherm at 232(o)c that is the melting point of andrographolide. The XRPD spectrum of the micropellets ascertained that the crystallinity of andrographolide was maintained. The relationship of the nature of the drug present in the micropellets related to release mechanisms is discussed. In conclusion, it can be said that andrographolide can be successfully incorporated into cross-linked micropellets of alginate without affecting its integrity or nature to deliver it as a bitterless monoherbal preparation.


Analysis of the Efficacy of Diet and Short-Term Probiotic Intervention on Depressive Symptoms in Patients after Bariatric Surgery: A Randomized Double-Blind Placebo Controlled Pilot Study.
(1) Background: studies have shown that some patients experience mental deterioration after bariatric surgery. (2) Methods: We examined whether the use of probiotics and improved eating habits can improve the mental health of people who suffered from mood disorders after bariatric surgery. We also analyzed patients' mental states, eating habits and microbiota. (3) Results: Depressive symptoms were observed in 45% of 200 bariatric patients. After 5 weeks, we noted an improvement in patients' mental functioning (reduction in BDI and HRSD), but it was not related to the probiotic used. The consumption of vegetables and whole grain cereals increased (DQI-I adequacy), the consumption of simple sugars and SFA decreased (moderation DQI-I), and the consumption of monounsaturated fatty acids increased it. In the feces of patients after RYGB, there was a significantly higher abundance of two members of the Muribaculaceae family, namely Veillonella and Roseburia, while those after SG had more Christensenellaceae R-7 group, Subdoligranulum, Oscillibacter, and UCG-005. (4) Conclusions: the noted differences in the composition of the gut microbiota (RYGB vs. SG) may be one of the determinants of the proper functioning of the gut-brain microbiota axis, although there is currently a need for further research into this topic using a larger group of patients and different probiotic doses.


Identification of reference genes for RT-qPCR expression analysis in Arabidopsis and tomato seeds.
Quantifying gene expression levels is an important research tool to understand biological systems. Reverse transcription-quantitative real-time PCR (RT-qPCR) is the preferred method for targeted gene expression measurements because of its sensitivity and reproducibility. However, normalization, necessary to correct for sample input and reverse transcriptase efficiency, is a crucial step to obtain reliable RT-qPCR results. Stably expressed genes (i.e. genes whose expression is not affected by the treatment or developmental stage under study) are indispensable for accurate normalization of RT-qPCR experiments. Lack of accurate normalization could affect the results and may lead to false conclusions. Since transcriptomes of seeds are different from other plant tissues, we aimed to identify reference genes specifically for RT-qPCR analyses in seeds of two important seed model species, i.e. Arabidopsis and tomato. We mined Arabidopsis seed microarray data to identify stably expressed genes and analyzed these together with putative reference genes from other sources. In total, the expression stability of 24 putative reference genes was validated by RT-qPCR in Arabidopsis seed samples. For tomato, we lacked transcriptome data sets of seeds and therefore we tested the tomato homologs of the reference genes found for Arabidopsis seeds. In conclusion, we identified 14 Arabidopsis and nine tomato reference genes. This provides a valuable resource for accurate normalization of gene expression experiments in seed research for two important seed model species.


KDM6A Depletion in Breast Epithelial Cells Leads to Reduced Sensitivity to Anticancer Agents and Increased TGFβ Activity.
KDM6A, an X chromosome-linked histone lysine demethylase, was reported to be frequently mutated in many tumor types including breast and bladder cancer. However, the functional role of KDM6A is not fully understood. Using MCF10A as a model of non-tumorigenic epithelial breast cells, we found that silencing KDM6A promoted cell migration and transformation demonstrated by the formation of tumor-like acini in three-dimensional culture. KDM6A loss reduced the sensitivity of MCF10A cells to therapeutic agents commonly used to treat patients with triple-negative breast cancer and also induced TGFβ extracellular secretion leading to suppressed expression of cytotoxic genes in normal human CD8+ T cells in vitro. Interestingly, when cells were treated with TGFβ, de novo synthesis of KDM6A protein was suppressed while TGFB1 transcription was enhanced, indicating a TGFβ/KDM6A-negative regulatory axis. Furthermore, both KDM6A deficiency and TGFβ treatment promoted disorganized acinar structures in three-dimensional culture, as well as transcriptional profiles associated with epithelial-to-mesenchymal transition and metastasis, suggesting KDM6A depletion and TGFβ drive tumor progression. Our study provides the preclinical rationale for evaluating KDM6A and TGFβ in breast tumor samples as predictors for response to chemo and immunotherapy, informing personalized therapy based on these findings.


[Pregnancy and HIV infection: the need for routine serological evaluation].
To determine the frequency of pregnancies among HIV-infected women in a sanitary area. To evaluate the proportion of women not receiving anti-retroviral treatment to decrease vertical transmission and the reasons why this treatment was not administered. Point prevalence study performed on all women followed for 1997 at the HIV Infection Unit in a 360-bed hospital. The following variables were obtained: social class, civil status and place of residence, risk factors for HIV infection, obstetric antecedents (pregnancies, number of term pregnancies, living newborns) as well as prescription or not of anti-retroviral therapy during pregnancy. Out of 85 women included in the study, 51 (60%) reported to have had a pregnancy and 17 of these (33%) had interrupted the pregnancy at some time. No significant differences were found between pregnancy or abortion and the analyzed socio-demographic variables or risk factors for HIV infection. Only 12% of women with a full length pregnancy received anti-retroviral therapy. Of women with term pregnancy who were not treated, most (63%) did not know they were infected before delivery and an additional 10% refused therapy. Forty-four percent of women with children continued with pregnancy despite knowing they were infected. Vertical transmission occurred in a 13% of cases in which no therapy was instituted and in no case in which zidovudine was administered during pregnancy. The frequency of pregnancies among HIV-infected women is high in our area and a substantial number of women do not know they are infected. These data support the serological study to HIV in all pregnant women and the necessity of a higher level of information in order that the seropositive women be aware of the responsibility she takes when she decides to go on with her pregnancy.


Idiopathic membranous nephropathy: diagnosis and treatment.
Idiopathic membranous nephropathy is still the most common glomerular disease associated with nephrotic syndrome. The greater the proteinuria, the greater the long-term risk for renal failure. In addition, patients who have membranous nephropathy with nephrotic syndrome have significant morbidity and mortality, in particular related to thromboembolic and cardiovascular complications. There is no specific treatment for membranous nephropathy. Supportive care with the use of diuretics and angiotensin-converting enzyme inhibitors in combination with angiotensin II receptor blocker is recommended, but these agents have only a limited effect. Immunosuppressive treatment options include the use of corticosteroids, alkylating agents, cyclosporin A, tacrolimus, and mycophenolate mofetil, but their use is controversial, not all have been shown to be effective, and their use can be associated with significant adverse effects. This has resulted in relatively small improvement in the prognosis of membranous nephropathy in the past 30 yr, with up to 40% of patients eventually reaching end-stage renal failure. Agents that offer more complete response rates with lower adverse effects are truly needed. Recent data suggest that B cells play a key role in the pathogenesis of a number of autoimmune diseases including membranous nephropathy and that selective depletion of B cells in humans may be beneficial in preventing the production of pathogenic immunoglobulins and subsequent renal injury.


Animal models for transdermal drug delivery.
The purpose of this investigation was to compare the permeation characteristics of two different compounds (extremely polar and nonpolar), i.e., tritium-labeled water (W) and 14C-labeled 7-hydroxycoumarin (7-OHC), among 16 different species, including human skin. Snake skin exhibited highest permeability for both W and 7-OHC. Permeability and lag time values of W and 7-OHC across Brattleboro rat and hairless guinea pig are very close to human skin. Skin thickness in micrometers (full thickness, epidermis and stratum corneum, and stratum corneum), and number of hair follicles present in each cm2 were determined for each species. There was no relationship between number of hair follicles and permeability values for both W and 7-OHC. The skin thickness (full) was related to the relative permeability values of W, whereas for 7-OHC it was not.


Long-term outcome in patients with prosthetic valve endocarditis: results from a single center in Turkey.
Prosthetic valve endocarditis (PVE) is associated with increased mortality and morbidity. Information regarding the long-term outcome of PVE is scarce in Turkey. The aim of this study was to evaluate long-term mortality rates of PVE and identify predictors of mortality in these patients. From January 2008 through August 2013, 44 patients (25 male, 19 female; mean age 49.3±12.1 years) who received a definitive diagnosis of PVE enrolled in the study.Median follow-up period was 23 months. Survival status was assessed for each patient by reviewing charts and making contact by phone. Cox regression analysis was used to evaluate outcome predictors. The mitral valve was the most commonly affected valve and Staphylococcus aureus the most prevalent microorganism. Fourteen patients (32%) underwent surgery, 7 of whom underwent early surgery. Overall mortality and in-hospital mortality rates were 39% (n=17) and 25% (n=11), respectively. In multivariate analysis, NYHA classification >2 (hazard ratio [HR] 3.7; 95% confidence interval [CI], 1.16-11.8; p=0.03), early-onset PVE (HR 4.23; 95% CI, 1.1-16.42; p=0.04), vegetation size ≥10 mm (HR 3.94; 95% CI, 1.1-14.75; p=0.04), and heart failure (HR 4.18; 95% CI, 1.36-12.8; p=0.01) were found to be independent predictors of mortality. Our findings suggest that PVE is associated with increased long-term mortality rates. Poor functional status, early-onset PVE, heart failure, and vegetation size are independent predictors of survival in patients with PVE.


Catecholamine-induced suppression of interleukin-1 production.
Recent evidence indicates that stress can suppress immune responses and thus increase the severity of viral and neoplastic diseases. Although, the mechanisms for stress-induced modulation of immunologic competence are unclear, neuroendocrine hormones are thought to be involved. A direct suppressive effect could result from the action of neuroendocrine hormones on lymphokine and monokine release. The central role of interleukin-1 (IL-1) in regulating cellular immune responses to infection and neoplasms stimulated the present study evaluating the effects of neuroendocrine hormones on IL-1 production. Norepinephrine and epinephrine inhibited the capacity of gamma interferon and lipopolysaccharide to stimulate IL-1 production from mouse peritoneal macrophages. Moreover, when intracellular and extracellular levels of IL-1 were quantitated, the studies demonstrated a catecholamine-mediated block in IL-1 synthesis without effect on its release. We also observed that exogenous cyclic AMP (cAMP) administered to mouse macrophages suppressed IL-1 production. This, coupled with the capacity of norepinephrine and epinephrine to enhance intracellular cAMP levels in macrophages, strongly suggested that the catecholamine-induced suppression of IL-1 production may be mediated by elevated intracellular cAMP levels. These findings demonstrate that selected stress-related neuroendocrine hormones can modulate IL-1 production by macrophages and further support the hypothesis that alteration of macrophage function by neuropeptides and neurohormones is a significant feature of stress-induced enhancement of viral and neoplastic disease.


Role of peroxisome proliferator-activated receptor-alpha in hepatobiliary injury induced by ammonium perfluorooctanoate in mouse liver.
Peroxisome proliferator-activated receptor-alpha (PPARalpha) has been suggested to protect against chemically induced hepatobiliary injuries in rodents. This function could mask the potential toxicities of perfluorooctanoic acid (PFOA) that is an emerging environmental contaminant and a weak ligand of PPARalpha. However its function has not been clarified. In this study, PFOA was found to elicit hepatocyte and bile duct injuries in Pparalpha-null mice after 4 wk treatment with PFOA ammonium salt (0, 12.5, 25, 50 micromol/kg/d, gavage). In wild-type mice, PFOA caused major hepatocellular damage dose-dependently and minor cholangiopathy observed only at 25 and 50 micromol/kg. In treated Pparalpha-null mice, PFOA produced marked fat accumulation, severe cholangiopathy, hepatocellular damage and apoptotic cells especially in bile ducts. Oxidative stress was also increased 4-fold at 50 micromol/kg and TNF-alpha mRNA was upregulated more than 3-fold at 25 micromol/kg in Pparalpha-null mice. Biliary bile acid/phospholipid ratios were higher in Pparalpha-null mice than in wild-type mice. Results from these studies suggest that PPARalpha is protective against PFOA and have a critical role in drug induced hepatobiliary injury.


Expression of Mycobacteriophage Ms6 lysis genes is driven by two sigma(70)-like promoters and is dependent on a transcription termination signal present in the leader RNA.
A mycobacteriophage Ms6 strong promoter region (P(lys)) was isolated by using transcriptional fusions with the lacZ reporter gene. Two tandem sigma(70)-like promoter sequences (P1 and P2) were found in this region. DNA sequencing of the promoter downstream region revealed a 214-bp leader sequence followed by five adjacent coding regions of 231 bp (ORF1), 1,152 bp (ORF2), 996 bp (ORF3), 231 bp (ORF4), and 372 (ORF5). ORF1 has the potential to encode a 77-amino-acid protein which revealed similarity to mycobacteriophage TM4 gp90, a predicted protein with unknown function. ORF2 encodes a 384-amino-acid protein which is related to several bacteriophage amidases. This protein induced cell lysis upon addition of chloroform, confirming its mureinolytic activity. ORF3 encodes a 332-amino-acid protein which is related to TM4 gp30, a protein with sequence similarity to amidases. ORF4 encodes a 77-amino-acid holin-like protein with significant similarity to the holin of Lactococcus lactis r1t bacteriophage. ORF5 encodes a 124-amino-acid protein which is related to mycobacteriophage L5 gp30, a protein with unknown function. These data indicate that the promoter region P(lys) drives the transcription of the Ms6 lysis genes. An intrinsic transcription termination signal was identified in the leader sequence. Experiments using lacZ fusions showed that beta-galactosidase synthesis is inhibited when this transcription termination signal is present in the leader sequence. In conclusion, mycobacteriophage Ms6 cell lysis genes are expressed by their own promoter region, independently of virion structure and assembly protein genes. Moreover, an antitermination mechanism might be involved in their transcription regulation.


Histopathology of cutaneous reaction to granulocyte colony-stimulating factor: another pseudomalignancy.
Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth factor (HGF) with many applications in cancer therapy. The most important applications are reduction in the incidence of febrile neutropenia, acceleration of neutrophil recovery after chemotherapy or bone marrow transplantation, and mobilization of progenitor cells. Many cutaneous adverse reactions associated with HGF have been reported in recent years, including injection site reactions, pyoderma gangrenosum, Sweet's syndrome, cutaneous leucocytoclastic vasculitis, and widespread folliculitis. The presence of large histiocytes on the dermis between collagen bundles has been proposed as a characteristic histopathologic finding in cutaneous eruptions secondary to granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. We report on a patient with a high-risk ductal infiltrating carcinoma of the breast who received high-dose chemotherapy (HDC) with peripheral blood progenitor cell (PBPC) rescue. The patient received G-CSF after PBPC for a faster granulocyte recovery. She developed a cutaneous eruption located on back, buttocks, axillae, groin and sites where electrocardiography electrodes had been placed. From the histopathological point of view, the eruption was characterized by the presence of numerous large, atypical histiocytes in the dermis with several mitotic figures, mimicking involvement of the dermis by a malignant process.


Weed spatial distribution as a function of soil properties in two distinct environments of the Brazilian semi-arid region.
The present study aimed was to evaluate the spatial variability of weed species by means of phytosociological parameters and their correlations with the physical-chemical soil properties, under semiarid climate conditions. Weed phytosociology and soil characterization were carried out in two areas one newly deforested area covering 8.86 ha, and one experimental agricultural area covering 24.7 ha; both in the semi-arid region of Brazil. Weed and soil were sampled by following georeferenced grids in each area. Biomass and the total number of weed individuals, as well as soil properties, were mapped by the ordinary Kriging method. The predominant herbaceous plants in the newly deforested area were Hexasepalum teres and Digitaria insularis. The weed species that predominated in the agricultural area were Cyperus rotundus L., Euphorbia heterophylla L. and Herissantia Crispa (L.) Brizicky; the latter species outstanding for dry biomass (873.5g). Spatial dependence was observed for the predominant species, except for Digitaria insularis. The spatial distribution of these weeds was conditioned by soil K+ contents in both areas, and by sand content for the experimental agricultural area. Therefore, these two soil attributes resulted key factors for weed infestation in this semi-arid region.


Influence of combined sulfachloropyridazine sodium and zinc on enzyme activities and biogas production during anaerobic digestion of swine manure.
In order to study the influence of different concentrations of zinc and sulfachloropyridazine sodium (SCPS) on anaerobic digestion (AD) during biogas production, we determined the levels of urease, dehydrogenase activity, and volatile fatty acids (VFAs) in batch tests. The experiments were conducted in small AD devices at a temperature of 37 °C using swine manure and wheat straw as raw materials. Four digestion trials were performed using different zinc and SCPS contents: control digestion with no additives (CK), SCPS at 630 mg kg-1 dry weight (S), SCPS at 630 mg kg-1 with zinc at 500 mg kg-1 dry weight (SL), and SCPS at 630 mg kg-1 with zinc at 5,000 mg kg-1 dry weight (SH). The biogas accumulation under S was 1.7 times that with CK, while SL and SH produced 78% and 35% of that under S, respectively. Correlation analysis showed that the accumulated biogas was significantly negatively correlated (p < 0.05) with VFAs, and the urease activity was significantly negatively correlated (p < 0.01) with zinc and significantly positively correlated with VFAs (p < 0.05). The dehydrogenase activity was strongly correlated (p < 0.01) with the biogas accumulated during the AD of swine manure.


MATLAB-Based Algorithm and Software for Analysis of Wavy Collagen Fibers.
Knowledge of soft tissue fiber structure is necessary for accurate characterization and modeling of their mechanical response. Fiber configuration and structure informs both our understanding of healthy tissue physiology and of pathological processes resulting from diseased states. This study develops an automatic algorithm to simultaneously estimate fiber global orientation, abundance, and waviness in an investigated image. To our best knowledge, this is the first validated algorithm which can reliably separate fiber waviness from its global orientation for considerably wavy fibers. This is much needed feature for biological tissue characterization. The algorithm is based on incremental movement of local regions of interest (ROI) and analyzes two-dimensional images. Pixels belonging to the fiber are identified in the ROI, and ROI movement is determined according to local orientation of fiber within the ROI. The algorithm is validated with artificial images and ten images of porcine trachea containing wavy fibers. In each image, 80-120 fibers were tracked manually to serve as verification. The coefficient of determination R2 between curve lengths and histograms documenting the fiber waviness and global orientation were used as metrics for analysis. Verification-confirmed results were independent of image rotation and degree of fiber waviness, with curve length accuracy demonstrated to be below 1% of fiber curved length. Validation-confirmed median and interquartile range of R2, respectively, were 0.90 and 0.05 for curved length, 0.92 and 0.07 for waviness, and 0.96 and 0.04 for global orientation histograms. Software constructed from the proposed algorithm was able to track one fiber in about 1.1 s using a typical office computer. The proposed algorithm can reliably and accurately estimate fiber waviness, curve length, and global orientation simultaneously, moving beyond the limitations of prior methods.


Sex therapy and psychophysiological research.
The author discusses two areas of psychophysiological research that may be relevant to the diagnosis and treatment of sexual dysfunction: investigations of sexual arousal in men and studies of nocturnal penile tumescence. There has been no convincing evidence to date of a predictable, generalizable autonomic response pattern to sexual arousal, but neuroendocrine research in this area has shown promise. A close temporal relationship has been found between REM sleep and penile tumescence in normal men-deviations from this pattern may be related to sexual dysfunction. The author suggests that further research is necessary in these areas of potential diagnostic and prognostic significance.


Elevated 25-hydroxy and normal 1,25-dihydroxy cholecalciferol serum concentrations in a successfully-treated case of vitamin D3 toxicosis in a dog.
A 4-y old, 27 kg spayed female German Shepherd dog was observed to ingest one 1-oz package of a rodenticide containing cholecalciferol. An initial serum calcium concentration of 15.7 mg/dl was successfully reduced to normal during 10 d using calcitonin and prednisolone. During that time, the serum 25-hydroxy and 1,25-dihydroxy cholecalciferol concentrations ranged from 637 to 315 ng/ml (normal 32 +/- 6 ng/ml) and 64 to 29 pg/ml (normal 34 +/- 19 pg/ml), respectively. Serum mid-molecule parathyroid hormone concentrations (76 to 97 pcmol/L) were within the normal range (85-140 pcmol/L). These data indicate that hypercalcemia seen in dogs following field exposures to cholecalciferol-containing rodenticides may be associated with elevated 25-hydroxy rather than 1,25-dihydroxy cholecalciferol. Consequently, serum 25-hydroxy cholecalciferol concentrations may be the most conclusive method for diagnosing hypervitaminosis D3 toxicosis in the live dog.


Morphoregulation of avian beaks: comparative mapping of growth zone activities and morphological evolution.
Avian beak diversity is a classic example of morphological evolution. Recently, we showed that localized cell proliferation mediated by bone morphogenetic protein 4 (BMP4) can explain the different shapes of chicken and duck beaks (Wu et al. [2004] Science 305:1465). Here, we compare further growth activities among chicken (conical and slightly curved), duck (straight and long), and cockatiel (highly curved) developing beak primordia. We found differential growth activities among different facial prominences and within one prominence. The duck has a wider frontal nasal mass (FNM), and more sustained fibroblast growth factor 8 activity. The cockatiel has a thicker FNM that grows more vertically and a relatively reduced mandibular prominence. In each prominence the number, size, and position of localized growth zones can vary: it is positioned more rostrally in the duck and more posteriorly in the cockatiel FNM, correlating with beak curvature. BMP4 is enriched in these localized growth zones. When BMP activity is experimentally altered in all prominences, beak size was enlarged or reduced proportionally. When only specific prominences were altered, the prototypic conical shaped chicken beaks were converted into an array of beak shapes mimicking those in nature. These results suggest that the size of beaks can be modulated by the overall activity of the BMP pathway, which mediates the growth. The shape of the beaks can be fine-tuned by localized BMP activity, which mediates the range, level, and duration of locally enhanced growth. Implications of topobiology vs. molecular blueprint concepts in the Evo-Devo of avian beak forms are discussed.


[18F]Fluorodeoxyglucose (FDG)-Positron Emission Tomography (PET)/Computed Tomography (CT) in Suspected Recurrent Breast Cancer: A Prospective Comparative Study of Dual-Time-Point FDG-PET/CT, Contrast-Enhanced CT, and Bone Scintigraphy.
To prospectively investigate the diagnostic accuracy of [(18)F]fluorodeoxyglucose (FDG)-positron emission tomography (PET)/computed tomography (CT) with dual-time-point imaging, contrast-enhanced CT (ceCT), and bone scintigraphy (BS) in patients with suspected breast cancer recurrence. One hundred women with suspected recurrence of breast cancer underwent 1-hour and 3-hour FDG-PET/CT, ceCT, and BS within approximately 10 days. The study was powered to estimate the precision of the individual imaging tests. Images were visually interpreted using a four-point assessment scale, and readers were blinded to other test results. The reference standard was biopsy along with treatment decisions and clinical follow-up (median, 17 months). FDG-PET/CT resulted in no false negatives and fewer false positives than the other imaging techniques. Accuracy of results were similar for 1-hour and 3-hour FDG-PET/CT. For distant recurrence, the area under the receiver operating curve was 0.99 (95% CI, 0.97 to 1) for FDG-PET/CT, 0.84 (95% CI, 0.73 to 0.94) for ceCT, and 0.86 (95% CI, 0.77 to 0.94) for the combined ceCT+BS. Of 100 patients, 22 (22%) were verified with distant recurrence, and 18 of these had bone involvement. Nineteen patients (19%) had local recurrence only. In exploratory analyses, diagnostic accuracy of FDG-PET/CT was better than ceCT alone or ceCT combined with BS in diagnosing distant, bone, and local recurrence, shown by a greater area under the receiver operating curve and higher sensitivity, specificity, and superior likelihood ratios. FDG-PET/CT was accurate in diagnosing recurrence in breast cancer patients. It allowed for distant recurrence to be correctly ruled out and resulted in only a small number of false-positive cases. Exploratory findings suggest that FDG-PET/CT has greater accuracy than conventional imaging technologies in this patient group.


The Association Between Trait Gratitude and Self-Reported Sleep Quality Is Mediated by Depressive Mood State.
It has been shown that higher levels of trait gratitude are associated with better self-reported sleep quality, possibly due to differences in presleep cognitions. However previous studies have not taken into account the role of depressive symptoms in this relationship. In this study, 88 nonclinical 18-29-year-olds completed the Gratitude Resentment and Appreciation Test (GRAT) as a measure of trait gratitude. The Glasgow Content of Thought Inventory (GCTI) was used to measure the intrusiveness of cognitions prior to sleep onset, the Motivation and Energy Inventory (MEI) assessed daytime fatigue, and the Pittsburgh Sleep Quality Index (PSQI) was used to assess self-reported sleep quality. The BDI-II assessed self-reported depressive symptoms. Consistent with previous work, GRAT scores were positively associated with higher daytime energy and greater number of hours of sleep per night. Importantly, however, we further observed that depressive symptoms mediated the relationships between gratitude scores and sleep metrics. Depressive mood state appears to mediate the association between gratitude and self-reported sleep quality metrics. We suggest, as one plausible model of these phenomena, that highly grateful individuals have lower symptoms of depression, which in turn leads to fewer presleep worries, resulting in better perceived sleep quality. Future work should aim to disentangle the causal nature of these relationships in order to better understand how these important variables interact.


Metabolic programming in the immediate postnatal life.
The metabolic programming effects of nutritional modifications in the immediate postnatal life are increasingly recognized to independently contribute to the development of metabolic syndrome in later life. Adjustment of litter size in rodents has been used to induce either under- or overnourishment in the immediate postnatal life of the offspring. While undernourishment led to growth retardation in the offspring, overnourishment produced increased body weight gains, hyperinsulinemia and hyperleptinemia. Overnourishment during the suckling period induced several adaptations in the energy circuitry in the hypothalamus of the offspring predisposing them for the onset of obesity later in life. Another approach for a nutritional modification in the immediate postnatal period is the artificial rearing of newborn rat pups on a high-carbohydrate (HC) milk formula without changes in the total calorie availability. Hyperinsulinemia, immediately evident in the HC pups, persisted in the post-weaning period even after withdrawal of the HC milk. Significant alterations in pancreatic islets supported chronic hyperinsulinemia in the HC rats. Alterations in the gene expression of hypothalamic neuropeptides predisposing to hyperphagia were evident during the period of the HC dietary modification. The persistence of these hypothalamic adaptations supported the obese phenotype in adult HC rats. A transgenerational effect gave rise to the development of chronic hyperinsulinemia and adult-onset obesity in the offspring of the HC female rats. Other studies have shown that lactation by a diabetic, obese or malnourished mother resulted in predisposition for the onset of metabolic disorders in the offspring. These observations from animal studies on the metabolic programming effects due to altered nutritional experiences in the immediate postnatal life strongly suggest that altered feeding practices for infants (formula feeding and early introduction of infant foods) could contribute to the rising incidence of overweight/obesity in children and adults.


A false-discovery-rate-based loss framework for selection of interactions.
Interaction effects have been consistently found important in explaining the variation in outcomes in many scientific research fields. Yet, in practice, variable selection including interactions is complicated due to the limited sample size, conflicting philosophies regarding model interpretability, and accompanying amplified multiple-testing problems. The lack of statistically sound algorithms for automatic variable selection with interactions has discouraged activities in exploring important interaction effects. In this article, we investigated issues of selecting interactions from three aspects: (1) What is the model space to be searched? (2) How is the hypothesis-testing performed? (3) How to address the multiple-testing issue? We propose loss functions and corresponding decision rules that control FDR in a Bayesian context. Properties of the decision rules are discussed and their performance in terms of power and FDR is compared through simulations. Methods are illustrated on data from a colorectal cancer study assessing the chemotherapy treatments and data from a diffuse large-B-cell lymphoma study assessing the prognostic effect of gene expressions.


Serratia marcescens surgical wound infection following breast reconstruction.
Surgical wound infections due to gram-negative bacilli have been rarely reported following breast implant surgery. From April to November 1989, four patients from one plastic surgeon's practice developed Serratia marcescens surgical wound infection (SWI) following breast reconstruction procedures with implantation of six expandable mammary implants. All six implants were removed for unabated S. marcescens SWI. Symptoms developed 13-161 days (median, 66 days) after surgery. When compared with nonexpandable silicone breast implants used during the period November 1, 1988, to October 31, 1989, expandable implants were associated with a greater risk of S. marcescens SWI (4/10 versus 0/11 patients, p = 0.04). Epidemiologic studies revealed that infection was associated with saline expansion of the implants performed in the surgeon's office. S. marcescens was cultured from a bag of commercial saline used on at least two of the four patients with SWI; the isolate from the saline and the three available patient isolates had identical serotype (O-undetermined:H4) and antimicrobial susceptibility patterns. Review of office procedures revealed that hands were not routinely washed before and aseptic technique was not used during the expansion procedure. Cultures of unopened bags of saline and an unused expandable implant were sterile. We hypothesize that multiple use of saline bags and nonsterile expansion technique extrinsically contaminated saline solutions and resulted in implant and/or surgical site infection. This investigation underscores the importance of avoiding multiple use of solutions intended for single use and of using aseptic technique when manipulating prosthetic devices.


An endocrine approach to the control of epidermal growth: serum-free cultivation of human keratinocytes.
Human keratinocytes, derived from the skin of newborns and of adults, were grown in the complete absence of serum, in a hormone-supplemented medium on fibronectin-coated cell culture dishes at low seed density. The cell culture medium consisted of Medium 199 containing epidermal growth factor, triiodothyronine, hydrocortisone, Cohn fraction IV, insulin, transferrin, bovine brain extract, and trace elements. Removal of the brain extract from the hormone supplement had a greater negative impact on proliferation of the keratinocyte cultures than did the removal of epidermal growth factor, hydrocortisone, and triiodothyronine or Cohn fraction IV. The growth of keratinocytes in this hormone-supplemented medium suggests that control of keratinocyte growth depends in large part on endocrine stimulation by other body organs, including the brain.