Alfaxalone or ketamine-medetomidine in cats undergoing ovariohysterectomy: a comparison of intra-operative parameters and post-operative pain.
To compare post-operative pain in cats after alfaxalone or ketamine- medetomidine anaesthesia for ovariohysterectomy (OHE) and physiologic parameters during and after surgery. Prospective 'blinded' randomized clinical study. Twenty-one healthy cats. Cats were assigned randomly into two groups: Group A, anaesthesia was induced and maintained with alfaxalone [5 mg kg(-1) intravenously (IV) followed by boli (2 mg kg(-1) IV); Group MK, induction with ketamine (5 mg kg(-1) IV) after medetomidine (30 μg kg(-1) intramuscularly (IM)], and maintenance with ketamine (2 mg kg(-1) IV). Meloxicam (0.2 mg kg(-1) IV) was administered after surgery. Basic physiological data were collected. At time T = -2, 0, 0.5, 1, 2, 4, 6, 8, 12, 16, 20, and 24 hours post-operatively pain was assessed by three methods, a composite pain scale (CPS; 0-24 points), a visual analogue scale (VAS 0-100 mm), and a mechanical wound threshold (MWT) device. Butorphanol (0.2 mg kg(-1) IM) was administered if CPS was scored ≥13. Data were analyzed using a general linear model, Kruskal-Wallis analyses, Bonferroni-Dunn test, unpaired t-test and Fisher's exact test as relevant. Significance was set at p < 0.05. VASs were significantly higher at 0.5, 1, 2, 4, and 20 hours in group A; MWT values were significantly higher at 8 and 12 hours in group MK. Post-operative MWT decreased significantly compared to baseline in both groups. There was no difference in CPS at any time point. Five cats required rescue analgesia (four in A; one in MK). Anaesthesia with ketamine-medetomidine was found to provide better post-surgical analgesia than alfaxalone in cats undergoing OHE; however, primary hyperalgesia developed in both groups. Alfaxalone is suitable for induction and maintenance of anaesthesia in cats undergoing OHE, but administration of additional sedative and analgesic drugs is highly recommended.


Characterization of a triad of genes in cyanophage S-2L sufficient to replace adenine by 2-aminoadenine in bacterial DNA.
Cyanophage S-2L is known to profoundly alter the biophysical properties of its DNA by replacing all adenines (A) with 2-aminoadenines (Z), which still pair with thymines but with a triple hydrogen bond. It was recently demonstrated that a homologue of adenylosuccinate synthetase (PurZ) and a dATP triphosphohydrolase (DatZ) are two important pieces of the metabolism of 2-aminoadenine, participating in the synthesis of ZTGC-DNA. Here, we determine that S-2L PurZ can use either dATP or ATP as a source of energy, thereby also depleting the pool of nucleotides in dATP. Furthermore, we identify a conserved gene (mazZ) located between purZ and datZ genes in S-2L and related phage genomes. We show that it encodes a (d)GTP-specific diphosphohydrolase, thereby providing the substrate of PurZ in the 2-aminoadenine synthesis pathway. High-resolution crystal structures of S-2L PurZ and MazZ with their respective substrates provide a rationale for their specificities. The Z-cluster made of these three genes - datZ, mazZ and purZ - was expressed in E. coli, resulting in a successful incorporation of 2-aminoadenine in the bacterial chromosomal and plasmidic DNA. This work opens the possibility to study synthetic organisms containing ZTGC-DNA.


Factors influencing the utilisation of health services by rural men.
This research identified the barriers and enablers that influence the utilisation of health services by rural men in the Midwest region of Western Australia. The methodology was based on participatory action research, including qualitative assessments to determine the issues for a larger quantitative study. Four variables were identified as predictors for the use of health services: those who attended for preventive reasons; those not affected by seasonal work; men who thought a medical telephone line was not important; and those who did not consider privacy an important issue. Modification of health service delivery to men could potentially enhance appropriate utilisation of health services in rural areas.


Atherosclerosis, dementia, and Alzheimer disease in the Baltimore Longitudinal Study of Aging cohort.
Although it is now accepted that asymptomatic cerebral infarcts are an important cause of dementia in the elderly, the relationship between atherosclerosis per se and dementia is controversial. Specifically, it is unclear whether atherosclerosis can cause the neuritic plaques and neurofibrillary tangles that define Alzheimer neuropathology and whether atherosclerosis, a potentially reversible risk factor, can influence cognition independent of brain infarcts. We examined the relationship between systemic atherosclerosis, Alzheimer type pathology, and dementia in autopsies from 200 participants in the Baltimore Longitudinal Study of Aging, a prospective study of the effect of aging on cognition, 175 of whom had complete body autopsies. Using a quantitative analysis of atherosclerosis in the aorta, heart, and intracranial vessels, we found no relationship between the degree of atherosclerosis in any of these systems and the degree of Alzheimer type brain pathology. However, we found that the presence of intracranial but not coronary or aortic atherosclerosis significantly increased the odds of dementia, independent of cerebral infarction. Given the large number of individuals with intracranial atherosclerosis in this cohort (136/200), the population attributable risk of dementia related to intracranial atherosclerosis (independent of infarction) is substantial and potentially reversible. Atherosclerosis of the intracranial arteries is an independent and important risk factor for dementia, suggesting potentially reversible pathways unrelated to Alzheimer pathology and stroke through which vascular changes may influence dementia risk.


A stomach road or "Magenstrasse" for gastric emptying.
Gastric muscle contractions grind and mix solid/liquid meal within the stomach, and move it into the bowels at a controlled rate. Contractions are of two types: slow volume-reducing contractions of the proximal stomach (the fundus), and peristaltic contraction waves in the distal stomach (the antrum). Fundic squeeze maintains gastro-duodenal pressure difference to drive gastric emptying. Emptying is generally assumed to proceed from the antrum to the fundus, so that ingested drugs can take hours to enter the small intestines and activate. Antral contraction waves (ACW), in contrast, generate fluid motions that break down and mix gastric content. Using a computer model of the human stomach, we discover a new function of these contraction waves apart from grinding and mixing. In coordination with fundic contraction, antral contraction waves move liquid content from the fundus along a very narrow path to the duodenum through the center of the antrum. Using physiological data, we show that this gastric emptying "Magenstrasse" (stomach road) can funnel liquid gastric content from the farthest reaches of the fundus directly to the intestines within 10 min. Consequently, whereas drugs (tablets, capsules, liquid) released off the Magenstrasse may require hours to enter the duodenum, at low concentration, when released on the Magenstrasse the drug can enter the duodenum and activate within 10 min-at high concentration. This discovery might explain observed high variability in drug initiation time, and may have important implications to both drug delivery and digestion, as well as to other wall-driven emptying of elastic containers.


High-order UWB pulses scheme to generate multilevel modulation formats based on incoherent optical sources.
We present a high-order UWB pulses generator based on a microwave photonic filter which provides a set of positive and negative samples by using the slicing of an incoherent optical source and the phase inversion in a Mach-Zehnder modulator. The simple scalability and high reconfigurability of the system permit a better accomplishment of the FCC requirements. Moreover, the proposed scheme permits an easy adaptation to pulse amplitude modulation, bi phase modulation, pulse shape modulation and pulse position modulation. The flexibility of the scheme for being adaptable to multilevel modulation formats permits to increase the transmission bit rate by using hybrid modulation formats.


The transforming Streptococcus pneumoniae in the 21st century.
Streptococcus pneumoniae, an important pathogen causing sepsis, sinusitis, otitis media, bacterial meningitis and bacterial pneumonia, results in global morbidity and mortality each year. The burden of pneumococcal disease is highest in children and the elderly. Treatment of pneumococcal infection has been hampered by the complexity of the host immune response. In recent decades, the increase of S. pneumoniae strains' resistance to beta-lactam antibiotics and other classes of antimicrobials has made treatment even more complicated. Fortunately, the advent of heptavalent conjugate vaccine confers a high degree of protection against pneumococcal disease and colonization caused by vaccine serotype strains. After the introduction of conjugate pneumococcal vaccine, invasive pneumococcal disease caused by vaccine serotypes and antibiotic-resistant isolates has been reduced. However, naturally transformable pneumococci may escape vaccine-induced immunity by switching their capsular genes to non-vaccine serotypes. Development of cheaper, serotype-independent vaccines based on a combination of protein antigens should be pursued.


Composition-dependent formation of platinum silver nanowires.
The understanding of shape control of colloidal nanoparticles is still rather limited even after well over a decade of intensive research efforts. While surface capping agents can greatly influence the growth habit of nanocrystals in solution, the formation of certain morphology can hardly be understood based on both experimental data and simulations. Without a good understanding of the origins for shape formation, deterministic approaches to the synthesis of nanostructures can be hard to realize. In this paper, we describe the synthesis and formation of PtAg alloy nanowires in the presence of oleylamine and oleic acid through the oriented attachment. Transmission electron microscopy study shows the formation of wormlike nanowires occurs largely at the composition around Pt(50)Ag(50). Both Pt and Ag rich alloy nanostructures form sphere-like or faceted nanoparticles under the same reaction conditions. Density functional theory calculation is used to understand the interactions between the functional groups of capping agents and low index planes of PtAg alloys. The structural order of interfaces after collision between primary particles is obtained by molecular dynamic simulation. The results indicate that the formation of alloy nanowires is mostly driven by the interplay between the binding energy of capping agents on alloy surfaces and the diffusion of atoms at the interface upon the collision of primary nanoparticles.


Patterning and assembly of ciliature are independent processes in hypotrich ciliates.
Mirror-imaged doublets of the hypotrich ciliate Pleurotricha lanceolata were induced and analyzed with respect to the overall patterning (structural asymmetry and polarity) of the individual components of the ciliature. The overall pattern is arranged as a mirror image, but the individual components in the two halves of the doublet show the same organizational asymmetry. These data demonstrate the independence of the mechanisms for this kind of large-scale (global) patterning and control of assembly of the individual ciliary components.


Metabolic shift in the emergence of hyperinvasive pandemic meningococcal lineages.
Hyperinvasive lineages of Neisseria meningitidis, which persist despite extensive horizontal genetic exchange, are a major cause of meningitis and septicaemia worldwide. Over the past 50 years one such lineage of meningococci, known as serogroup A, clonal complex 5 (A:cc5), has caused three successive pandemics, including epidemics in sub-Saharan Africa. Although the principal antigens that invoke effective immunity have remained unchanged, distinct A:cc5 epidemic clones have nevertheless emerged. An analysis of whole genome sequence diversity among 153 A:cc5 isolates identified eleven genetic introgression events in the emergence of the epidemic clones, which primarily involved variants of core genes encoding metabolic processes. The acquired DNA was identical to that found over many years in other, unrelated, hyperinvasive meningococci, suggesting that the epidemic clones emerged by acquisition of pre-existing metabolic gene variants, rather than 'virulence' associated or antigen-encoding genes. This is consistent with mathematical models which predict the association of transmission fitness with the emergence and maintenance of virulence in recombining commensal organisms.


Characterization and differentiation of filamentous fungi based on Fatty Acid composition.
Cellular fatty acid composition of 100 different filamentous fungi, including oomycetes, zygomycetes, ascomycetes, basidiomycetes, and sterile mycelia, was analyzed to determine if they can be differentiated from one another on this basis and how minor variations in culture temperature and age affect this characteristic. Many fungi were found to possess the same fatty acids but produced different relative concentrations of each. Some fungi differed in both the fatty acids produced and in the relative concentrations of others. Multivariate discriminant analysis demonstrated that all of the species included in this study had significantly different (P < 0.001) fatty acid profiles. Each of the three phyla from which representative species were analyzed and the sterile forms had distinctive fatty acid profiles. Significant differences in fatty acid composition were also found at the intraspecific level. Both culture temperature and age affected fatty acid composition in the fungi examined, but when these factors were held constant, variance in fatty acid composition was not a problem and fungal fatty acid profiles could be differentiated statistically.


Body mass index cutoff points for evaluation of nutritional status in Brazilian children and adolescents.
To delineate a classification system, comprising reference curves and cutoff points, based on the distribution of body mass index (BMI) across a national reference population and designed for the assessment of the nutritional status of Brazilian children and adolescents. Data from 13,279 males and 12,823 females aged from 2 to 19 years, extracted from the National Nutrition and Health Survey dataset (1989), were used to construct a reference curve. The LMS method was employed to calculate the BMI curve parameters and polynomial functions were used to model these parameters against age. The cutoff values for classifying nutritional status as underweight, overweight and obese were expressed as centiles and BMI values equivalent to 17.5, 25 and 30 kg/m(2) at 20 years, respectively. Values for the L, M and S parameters were tabulated at 6-month intervals for each sex. Using these values, a graph was plotted with nine BMI distribution reference centiles. Cutoff values were presented that are equivalent to BMIs of 17.5, 25 and 30 kg/m(2) at the start of adulthood. The classification system presented here can be used for clinical and epidemiological assessments, it is methodologically similar to the majority of national curves that have been presented to date and, furthermore, it offers a definition of underweight.


Glucose metabolites in blood and adipose tissue of obese diabetic and non-diabetic subjects.
1. Glucose 6-phosphate, fructose 6-phosphate, fructose diphosphate, glycerol phosphate and uridine diphosphate glucose have been measured in human adipose tissue and blood from obese subjects under fed and fasting conditions and in obese diabetic and non-diabetic subjects before and after an oral glucose load (100 g). 2. Adipose tissue metabolites expressed as nmol/g wet weight correlated inversely with adipocyte diameter. 3. After fasting, fructose diphosphate and glycerol phosphate in adipose tissue decreased significantly. 4. The basal concentrations of metabolites in blood and adipose tissue were maintained at similar concentrations in diabetic and non-diabetic subjects despite very different blood glucose concentrations. 5. The significant increase in adipose tissue glucose 6-phosphate after the glucose load seen in the non-diabetic but not in the diabetic subjects suggest that glucose uptake is decreased in the diabetic adipocyte.


Alterations in medical interpretation during routine primary care.
Increasing numbers of patients require medical interpretation, yet few studies have examined its accuracy or effect on health outcomes. To understand how alterations in medical interpretation affect health care delivery to patients with limited English proficiency (LEP), we aimed to determine the frequency, type, and clinical significance of alterations. We focused on best-case encounters that involved trained, experienced interpreters interacting with established patients. We audio-recorded routine outpatient clinic visits in which a medical interpreter participated. Audiotapes were transcribed and translated into English. We identified and characterized alterations in interpretation and calculated their prevalence. In total, 38 patients, 16 interpreters, and 5 providers took part. Patients spoke Cantonese, Mandarin, Somali, Spanish, and Vietnamese, and received care for common chronic health conditions. Unlike previous methods that report numbers of alterations per interpreted encounter, we focused on alterations per utterance, which we defined as the unit of spoken content given to the interpreter to interpret. All alteration rates were calculated by dividing the number of alterations made during the encounter by the number of utterances for that encounter. We defined clinically significant changes as those with potential consequences for evaluation and treatment. We found that 31% of all utterances during a routine clinical encounter contained an alteration. Only 5% of alterations were clinically significant, with 1% having a positive effect and 4% having a negative effect on the clinical encounter. Even in a best case scenario, the rate of alteration remains substantial. Training interpreters and clinicians to address common patterns of alteration will markedly improve the quality of communication between providers and LEP patients.


Dictyostelium myosin IK is involved in the maintenance of cortical tension and affects motility and phagocytosis.
Dictyostelium discoideum myosin Ik (MyoK) is a novel type of myosin distinguished by a remarkable architecture. MyoK is related to class I myosins but lacks a cargo-binding tail domain and carries an insertion in a surface loop suggested to modulate motor velocity. This insertion shows similarity to a secondary actin-binding site present in the tail of some class I myosins, and indeed a GST-loop construct binds actin. Probably as a consequence, binding of MyoK to actin was not only ATP- but also salt-dependent. Moreover, as both binding sites reside within its motor domain and carry potential sites of regulation, MyoK might represent a new form of actin crosslinker. MyoK was distributed in the cytoplasm with a significant enrichment in dynamic regions of the cortex. Absence of MyoK resulted in a drop of cortical tension whereas overexpression led to significantly increased tension. Absence and overexpression of MyoK dramatically affected the cortical actin cytoskeleton and resulted in reduced initial rates of phagocytosis. Cells lacking MyoK showed excessive ruffling, mostly in the form of large lamellipodia, accompanied by a thicker basal actin cortex. At early stages of development, aggregation of myoK null cells was slowed due to reduced motility. Altogether, the data indicate a distinctive role for MyoK in the maintenance and dynamics of the cell cortex.


Increases in Perspective Embedding Increase Reading Time Even with Typical Text Presentation: Implications for the Reading of Literature.
Reading fiction is a major component of intellectual life, yet it has proven difficult to study experimentally. One aspect of literature that has recently come to light is perspective embedding ("she thought I left" embedding her perspective on "I left"), which seems to be a defining feature of fiction. Previous work (Whalen et al., 2012) has shown that increasing levels of embedment affects the time that it takes readers to read and understand short vignettes in a moving window paradigm. With increasing levels of embedment from 1 to 5, reading times in a moving window paradigm rose almost linearly. However, level 0 was as slow as 3-4. Accuracy on probe questions was relatively constant until dropping at the fifth level. Here, we assessed this effect in a more ecologically valid ("typical") reading paradigm, in which the entire vignette was visible at once, either for as long as desired (Experiment 1) or a fixed time (Experiment 2). In Experiment 1, reading times followed a pattern similar to that of the previous experiment, with some differences in absolute speed. Accuracy matched previous results: fairly consistent accuracy until a decline at level 5, indicating that both presentation methods allowed understanding. In Experiment 2, accuracy was somewhat reduced, perhaps because participants were less successful at allocating their attention than they were during the earlier experiment; however, the pattern was the same. It seems that literature does not, on average, use easiest reading level but rather uses a middle ground that challenges the reader, but not too much.


Selection of virus variants and emergence of virus escape mutants after immunization with an epitope vaccine.
In this report, we assessed the evolution of the cytotoxic T-lymphocyte (CTL) response induced by an epitope vaccine. In two macaques immunized with a mixture of lipopeptides derived from simian immunodeficiency virus (SIV) Nef and Gag proteins, CTL responses were directed against the same, single epitope of the Nef protein (amino acids 128 to 137) presenting an alanine at position 136 (Nef 128-137/136A). However, after 5 months of SIV infection, peripheral blood mononuclear cells from both macaques lost their ability to be stimulated by autologous SIV-infected cells while still retaining their capacity to generate cytotoxic responses after specific Nef 128-137/136A peptide stimulation. The sequences of the pathogenic viral isolate used for the challenge showed a mixture of several variants. Within the Nef epitopic sequence from amino acids 128 to 137, 82% of viral variants expressed the epitopic peptide Nef 128-137/136A; the remaining variants presented a threonine at position 136 (Nef 128-137/136T). In contrast, sequence analysis of cloned proviral DNA obtained from both macaques 5 months after SIV challenge showed a different pattern of quasi-species variants; 100% of clones presented a threonine at position 136 (Nef 128-137/136T), suggesting the disappearance of viral variants with an alanine at this position under antiviral pressure exerted by Nef 128-137/136A-specific CTLs. In addition, 12 months after SIV challenge, six of eight clones from one macaque presented a glutamic acid at position 131 (Nef 128-137/131E+136T), which was not found in the infecting isolate. Furthermore, CTLs generated very early after SIV challenge were able to lyse cells sensitized with the Nef 128-137/136A epitope. In contrast, lysis was significantly less effective or even did not occur when either the selected peptide Nef 128-137/136T or the escape variant peptide Nef 128-137/131E+136T was used in a target cell sensitization assay. Dose analysis of peptides used to sensitize target cells as well as a major histocompatibility complex (MHC)-peptide stability assay suggested that the selected peptide Nef 128-137/136T has an altered capacity to bind to the MHC. These data suggest that CTL pressure leads to the selection of viral variants and to the emergence of escape mutants and supports the fact that immunization should elicit broad CTL responses.


Influenza A Virus Infection Induces Viral and Cellular Defective Ribosomal Products Encoded by Alternative Reading Frames.
The importance of antiviral CD8+ T cell recognition of alternative reading frame (ARF)-derived peptides is uncertain. In this study, we describe an epitope (NS1-ARF21-8) present in a predicted 14-residue peptide encoded by the +1 register of NS1 mRNA in the influenza A virus (IAV). NS1-ARF21-8 elicits a robust, highly functional CD8+ T cell response in IAV-infected BALB/c mice. NS1-ARF21-8 is presented from unspliced NS mRNA, likely from downstream initiation on a Met residue that comprises the P1 position of NS1-ARF21-8 Derived from a 14-residue peptide with no apparent biological function and negligible impacts on IAV infection, infectivity, and pathogenicity, NS1-ARF21-8 provides a clear demonstration of how immunosurveillance exploits natural errors in protein translation to provide antiviral immunity. We further show that IAV infection enhances a model cellular ARF translation, which potentially has important implications for virus-induced autoimmunity.


Knockdown of Drosha in human alveolar type II cells alters expression of SP-A in culture: a pilot study.
Human surfactant protein A (SP-A) plays an important role in surfactant metabolism and lung innate immunity. SP-A is synthesized and secreted by alveolar type II (ATII) cells, one of the two cell types of the distal lung epithelium (ATII and ATI). We have shown that miRNA interactions with sequence polymorphisms on the SP-A mRNA 3'UTRs mediate differential expression of SP-A1 and SP-A2 gene variants in vitro. In the present study, we describe a physiologically relevant model to study miRNA regulation of SP-A in human ATII. For these studies, we purified and cultured human ATII on an air-liquid interface matrix (A/L) or plastic wells without matrix (P). Gene expression analyses confirmed that cells cultured in A/L maintained the ATII phenotype for over 5 days, whereas P-cultured cells differentiated to ATI. When we transfected ATII with siRNAs to inhibit the expression of Drosha, a critical effector of miRNA maturation, the levels of SP-A mRNA and protein increased in a time dependent manner. We next characterized cultured ATII and ATI by studying expression of 1,066 human miRNAs using miRNA PCR arrays. We detected expression of >300 miRNAs with 24 miRNAs differentially expressed in ATII versus ATI, 12 of which predicted to bind SP-A 3'UTRs, indicating that these may be implicated in SP-A downregulation in ATI. Thus, miRNAs not only affect SP-A expression, but also may contribute to the maintenance of the ATII cell phenotype and/or the trans-differentiation of ATII to ATI cells, and may represent new molecular markers that distinguish ATII and ATI.


Genomic medicine at the heart of diabetes management.
Individual predisposition to type 2 diabetes is influenced by the combined effect of a constellation of genetic variants and a multitude of environmental exposures. Identification of the specific genetic variants involved, and the mechanisms through which they operate, provides a powerful approach for delivering biological insights that can drive translational benefit, one that is already widely exploited in the personalised management of monogenic and syndromic forms of diabetes. This commentary develops the argument that equivalent translational advances for more common forms of diabetes are unlikely to result solely from the ability to define more complete individual inventories of genetic risk and environmental exposure. They will also require identification of complex molecular signatures able to provide integrative, empirical, longitudinal readouts of disease progression. These signatures will track causal mechanisms and capture an individual's position within a complex spectrum of pathophysiological processes, thereby supporting personalised approaches to intervention and treatment. This is one of a series of commentaries under the banner '50 years forward', giving personal opinions on future perspectives in diabetes, to celebrate the 50th anniversary of Diabetologia (1965-2015).


Patient stratification and identification of adverse event correlations in the space of 1190 drug related adverse events.
New pharmacovigilance methods are needed as a consequence of the morbidity caused by drugs. We exploit fine-grained drug related adverse event information extracted by text mining from electronic medical records (EMRs) to stratify patients based on their adverse events and to determine adverse event co-occurrences. We analyzed the similarity of adverse event profiles of 2347 patients extracted from EMRs from a mental health center in Denmark. The patients were clustered based on their adverse event profiles and the similarities were presented as a network. The set of adverse events in each main patient cluster was evaluated. Co-occurrences of adverse events in patients (p-value < 0.01) were identified and presented as well. We found that each cluster of patients typically had a most distinguishing adverse event. Examination of the co-occurrences of adverse events in patients led to the identification of potentially interesting adverse event correlations that may be further investigated as well as provide further patient stratification opportunities. We have demonstrated the feasibility of a novel approach in pharmacovigilance to stratify patients based on fine-grained adverse event profiles, which also makes it possible to identify adverse event correlations. Used on larger data sets, this data-driven method has the potential to reveal unknown patterns concerning adverse event occurrences.


Emergency room use of hypnosis.
Myths about hypnosis have interfered with its use in emergency settings. Specifically, included are myths about who induces the hypnotic state, the length of induction time, and the traumatized patient's inability to concentrate on a focal point. It is suggested, however, that altered states of awareness occur rapidly and spontaneously in the patient who has experienced acute trauma and/or pain. Two cases are presented that illustrate spontaneous trancelike states occurring in traumatic situations. The cases also show how the recognition of these altered states can facilitate the patient's care and treatment in an emergency setting.


Isolation, identification and function of a novel anti-HSV-1 protein from Grifola frondosa.
A novel antiviral protein was purified from an extract of Grifola frondosa fruiting bodies using a procedure that included 40% ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography, and designated GFAHP. This protein inhibited herpes simplex virus type 1 (HSV-1) replication in vitro with an IC(50) value of 4.1 microg/ml and a therapeutic index >29.3. Higher concentrations of GFAHP (125 and 500 microg/ml) also significantly reduced the severity of HSV-1 induced blepharitis, neovascularization, and stromal keratitis in a murine model. Topical administration of GFAHP to the mouse cornea resulted in a significant decrease in virus production (mean virus yields: 3.4log10PFU in the treated group and 4.19log10PFU in the control group). We proved that GFAHP directly inactivates HSV-1 while simultaneously inhibiting HSV-1 penetration into Vero cells. Gel electrophoresis showed that GFAHP had a molecular weight of 29.5 kDa. GFAHP was tryptic digested and analyzed from the PMF of matrix assisted desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) and nanoelectrospray ionization tandem mass spectrometry. The N-terminal sequence of GFAHP consisted of an 11 amino acid peptide, NH(2)-REQDNAPCGLN-COOH that did not match any known amino acid sequences, indicating that GFAHP is likely to be a novel antivirus protein. To our knowledge, this is the first report that characterizes an anti-HSV protein from G. frondosa.


Faraday current sensor with temperature monitoring.
We present a novel optical current sensor based on the Faraday effect that incorporates a temperature monitoring system. The monitoring element is a temperature-dependent birefringent plate placed at the Faraday sensor head. Measurement of the plate retardation permits compensation for the temperature dependence of the Verdet constant. Validation experiments are presented and discussed.


Expression of class III beta tubulin in non-small cell lung cancer is correlated with resistance to taxane chemotherapy.
This study determined the prevalence and the prognostic value of the expression of microtubule components in tumors of 19 patients with non small cell lung cancer receiving taxane-based regimens. Patient samples were stained with antibodies directed against total beta tubulin, classes I, II and III beta tubulin isotypes, delta2 alpha tubulin, tau protein, and the P-gp protein involved in the classical multidrug resistance phenotype. All tumors were stained with pan-beta tubulin antibody and class I tubulin isotype. A majority of the tumor samples expressed class II and class III, although the percentage of positive cells varied significantly between tumors. delta2 alpha tubulin, tau protein and Pgp protein were found in only one tumor sample each. Progression-free survival was shorter (41 days) in patients whose tumors expressed high levels of class III tubulin isotype in comparison to patients with low levels (288 days, p = 0.02). There were 2 responses to chemotherapy among 9 patients (22%) with high levels of class III tubulin vs. 6 among 10 patients (60%) with low levels of expression (Fisher exact test: p = 0.11). These data suggest that high expression of class III tubulin by tumor cells is associated with poor prognosis in patients with NSCLC receiving a taxane-based regimen.


In vitro maturation in sub fertile women with polycystic ovarian syndrome undergoing assisted reproduction.
Polycystic ovarian syndrome (PCOS) occurs in 4% to 7% of all women of reproductive age and 20% of women presenting with sub-fertility. A significant proportion of these women will ultimately need assisted reproductive techniques (ART). In women with PCOS, the supra-physiological doses of gonadotrophins used for controlled ovarian hyperstimulation (COH) often result in an exaggerated ovarian response characterised by the development of a large cohort of follicles of uneven quality, retrieval of immature oocytes, and increased risk of ovarian hyperstimulation syndrome. A potentially useful intervention for women with PCOS-related infertility involves earlier retrieval of immature oocytes followed by in vitro maturation (IVM). To compare live birth rates per woman following in vitro maturation (IVM) with conventional in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) for women with PCOS undergoing ART. We searched the Menstrual Disorders and Subfertility Group Specialised Register of controlled trials for any relevant trials from the title, abstract, or keyword sections. Following a search of electronic databases (MEDLINE in all languages) using Ovid software we also performed a manual search of the references of all retrieved articles, sought unpublished papers and abstracts submitted to international conferences, and contacted experts. In addition, we searched the National Institute of Clinical Excellence fertility assessment and treatment guidelines (NICE 2004) and handsearched reference lists of relevant systematic reviews and randomised trials. All randomised trials on the intention to perform IVM before IVF or ICSI and conventional IVF or ICSI for sub-fertile women with PCOS. We identified no studies that met the inclusion criteria. There were no trials suitable for inclusion in the review. There are no randomised controlled trials upon which to base any practice recommendations regarding IVM before IVF or ICSI for women with PCOS. There is an urgent need for randomised trials in this field.


Durability of continuous-flow left ventricular assist devices: a systematic review.
Left ventricular assist devices (LVADs) are becoming an increasingly viable alternative therapy for heart failure, either as a bridge to heart transplantation (BTT) or destination therapy (DT). The latter has become increasingly popular in recent years, in the face of a donor organ shortage and a rise in elderly patients ineligible for heart transplants. For these patients in particular, device durability is a key contributor to survival, morbidity, and quality of life. This systematic review aimed to assess the long-term durability of current continuous-flow LVADs. Six electronic databases were searched from their dates of inception to August 2014 for original studies reporting on patients receiving continuous-flow LVADs. LVAD failure was defined as device malfunction necessitating exchange or explantation, or causing patient mortality. Pooled averages were calculated for outcomes and rates of device failure were reconstructed from digitized graph curves using the software, WebPlotDigitizer v3.3. Twelve retrospective observational studies with a total of 5,471 patients were included for analysis. The mean duration of LVAD support was 504.7 (range, 303-568) days, and the overall weighted incidence of device failure was 3.9% (range, 1-11.3%). On average, pump thrombosis was the most common cause of device failure (50.5%), followed by lead or cable damage (21.7%), mechanical pump failure (11.6%), device-related infection (11.1%), and surgical complications from implantation (2.5%). Long-term device failure rates at 2-, 6-, 12-, 18- and 24-months post-implantation were 0.5%, 1.8%, 2.9%, 4.5% and 6.5%, respectively. With the expected rise in LVAD usage for end-stage heart failure, particularly as a DT, the steady minority of patients experiencing device failure is likely to increase. Further investigation is required into the incidence and mechanism of major causes of failure, as well as design improvements that may address these complications. There is currently a lack of guidelines and large randomized studies reporting on the etiology and outcomes of LVAD failure.


Effectiveness and content analysis of interventions to enhance medication adherence and blood pressure control in hypertension: A systematic review and meta-analysis.
The objective of this systematic review is to evaluate the effectiveness of medication adherence interventions on blood pressure control in hypertensive patients. In addition, we aim to explore what barriers and facilitators in the interventions may have been targeted and how these might be related to the effect size on blood pressure (BP). This review is a hypertension-specific update to the previous Cochrane Review by Nieuwlaat et al. ( 2014 ) on interventions to enhance medication adherence. A systematic literature search was carried out and two authors independently screened titles and abstracts for their eligibility for inclusion and independently extracted data from the selected studies and assessed the methodological quality using the Cochrane Collaboration Risk of Bias Tool. A meta-analysis was conducted and additionally, theoretical factors in interventions were identified using the Theoretical Domains Framework. The meta-analysis found a modest main effect of adherence interventions on SBP (MD -2.71 mm Hg, 95% CI -4.17 to -1.26) and DBP (MD -1.25 mm Hg, 95% CI -1.72 to -.79). However, there was substantial significant heterogeneity across both outcomes. A narrative review on adherence outcomes was conducted. In terms of the theoretical analysis, the relationship between the total number of times the domains were coded within an intervention and change of SBP (r = -.234, p = .335) and DBP was not significant (r = -.080, p = .732). Similarly, the relationship between the total number of times different domains were coded within an intervention and change of SBP (r = .080, p = .746) and DBP was not significant (r = -.188, p = .415). This review and meta-analysis of interventions documented significant but modest post-intervention improvements in BP outcomes among hypertensive patients. However, this is a tentative finding as substantial heterogeneity and potential biases were present. One of the greatest challenges of this review was assessing risk of bias, extracting sufficient data to calculate effect size and coding interventions with the amount of information provided in papers. It is imperative that future adherence research comprehensively reports methodology.


Mutation of luxS affects biofilm formation in Streptococcus mutans.
Quorum sensing is a bacterial mechanism for regulating gene expression in response to changes in population density. Many bacteria are capable of acyl-homoserine lactone-based or peptide-based intraspecies quorum sensing and luxS-dependent interspecies quorum sensing. While there is good evidence about the involvement of intraspecies quorum sensing in bacterial biofilm, little is known about the role of luxS in biofilm formation. In this study, we report for the first time that luxS-dependent quorum sensing is involved in biofilm formation of Streptococcus mutans. S. mutans is a major cariogenic bacterium in the multispecies bacterial biofilm commonly known as dental plaque. An ortholog of luxS for S. mutans was identified using the data available in the S. mutans genome project (http://www.genome.ou.edu/smutans.html). Using an assay developed for the detection of the LuxS-associated quorum sensing signal autoinducer 2 (AI-2), it was demonstrated that this ortholog was able to complement the luxS negative phenotype of Escherichia coli DH5alpha. It was also shown that AI-2 is indeed produced by S. mutans. AI-2 production is maximal during mid- to late-log growth in batch culture. Mutant strains devoid of the luxS gene were constructed and found to be defective in producing the AI-2 signal. There are also marked phenotypic differences between the wild type and the luxS mutants. Microscopic analysis of in vitro-grown biofilm structure revealed that the luxS mutant biofilms adopted a much more granular appearance, rather than the relatively smooth, confluent layer normally seen in the wild type. These results suggest that LuxS-dependent signal may play an important role in biofilm formation of S. mutans.


Effects of insecticides on the fluidity of mitochondrial membranes of the diamondback moth, Plutella xylostella, resistant and susceptible to avermectin.
The effects of various insecticides on the fluidity of mitochondrial membranes and cross-resistance were investigated in the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae) using strains that were both resistant and susceptible to avermectin. The resistant strain of P. xylostella, AV-R, developed 1078-fold resistance to avermetins with a high level of cross-resistance to the analogs of avermectins, ivermectin and emamectin benzoate. It had more than 1000 times greater resistance when compared with the avermectin-susceptible strain, XH-S. Mitochondrial membrane fluidity was measured by detecting fluorescence polarization using DPH (1,6-Diphenyl -1,3,5-hexatriene) as the fluorescence probe. Abamectin, emamectin benzoate, ivermectin, cypermethrin and fenvalerate decreased the fluidity of mitochondrial membranes in the XH-S strain at 25 degrees C. However, fipronil and acephate did not change the fluidity of mitochondrial membrane when the concentration of these insecticides was 1x10(-4) mol/L. Membrane fluidity increased as the temperature increased. The thermotropic effect on the polarization value of DPH increased as the insecticide concentration was increased. There was a significant difference of mitochondrial membrane fluidity between both XH-S and AV-R when temperature was less than 25 degrees C and no difference was observed when the temperature was more than 25 degrees C. The low-dose abamectin (0.11 mg/L) in vivo treatment caused a significant change of membrane fluidity in the XH-S strain and no change in the AV-R strain. However, a high-dose abamectin (11.86 mg/L) resulted in 100% mortality of the XH-S strain. In vivo treatment may cause a significant change of membrane fluidity in the AV-R strain.


Neurodevelopmental outcomes in infants requiring resuscitation in developing countries.
To determine whether resuscitation of infants who failed to develop effective breathing at birth increases survivors with neurodevelopmental impairment. Infants unresponsive to stimulation who received bag and mask ventilation at birth in a resuscitation trial and infants who did not require any resuscitation were randomized to early neurodevelopmental intervention or control groups. Infants were examined by trained neurodevelopmental evaluators masked to both their resuscitation history and intervention group. The 12-month neurodevelopmental outcome data for both resuscitated and non-resuscitated infants randomized to the control groups are reported. The study provided no evidence of a difference between the resuscitated infants (n = 86) and the non-resuscitated infants (n = 115) in the percentage of infants at 12 months with a Mental Developmental Index <85 on the Bayley Scales of Infant Development-II (primary outcome; 18% versus 12%; P = .22) and in other neurodevelopmental outcomes. Most infants who received resuscitation with bag and mask ventilation at birth have 12-month neurodevelopmental outcomes in the reference range. Longer follow-up is needed because of increased risk for neurodevelopmental impairments.


Marginal vitamin B-6 deficiency decreases plasma (n-3) and (n-6) PUFA concentrations in healthy men and women.
Previous animal studies showed that severe vitamin B-6 deficiency altered fatty acid profiles of tissue lipids, often with an increase of linoleic acid and a decrease of arachidonic acid. However, little is known about the extent to which vitamin B-6 deficiency affects human fatty acid profiles. The aim of this study was to determine the effects of marginal vitamin B-6 deficiency on fatty acid profiles in plasma, erythrocytes, and peripheral blood mononuclear cells (PBMC) of healthy adults fed a 28-d, low-vitamin B-6 diet. Healthy participants (n = 23) received a 2-d, controlled, vitamin B-6-adequate diet followed by a 28-d, vitamin B-6-restricted diet to induce a marginal deficiency. Plasma HDL and LDL cholesterol concentrations, FFA concentrations, and erythrocyte and PBMC membrane fatty acid compositions did not significantly change from baseline after the 28-d restriction. Plasma total arachidonic acid, EPA, and DHA concentrations decreased from (mean ± SD) 548 ± 96 to 490 ± 94 μmol/L, 37 ± 13 to 32 ± 13 μmol/L, and 121 ± 28 to 109 ± 28 μmol/L [positive false discovery rate (pFDR) adjusted P < 0.05], respectively. The total (n-6):(n-3) PUFA ratio in plasma exhibited a minor increase from 15.4 ± 2.8 to 16.6 ± 3.1 (pFDR adjusted P < 0.05). These data indicate that short-term vitamin B-6 restriction decreases plasma (n-3) and (n-6) PUFA concentrations and tends to increase the plasma (n-6):(n-3) PUFA ratio. Such changes in blood lipids may be associated with the elevated risk of cardiovascular disease in vitamin B-6 insufficiency.


Adaptation to statins restricts human tumour growth in Nude mice.
Statins have long been used as anti-hypercholesterolemia drugs, but numerous lines of evidence suggest that they may also bear anti-tumour potential. We have recently demonstrated that it was possible to isolate cancer cells adapted to growth in the continuous presence of lovastatin. These cells grew more slowly than the statin-sensitive cells of origin. In the present study, we compared the ability of both statin-sensitive and statin-resistant cells to give rise to tumours in Nude mice. HGT-1 human gastric cancer cells and L50 statin-resistant derivatives were injected subcutaneously into Nude mice and tumour growth was recorded. At the end of the experiment, tumours were recovered and marker proteins were analyzed by western blotting, RT-PCR and immunohistochemistry. L50 tumours grew more slowly, showed a strong decrease in cyclin B1, over-expressed collagen IV, and had reduced laminin 332, VEGF and CD34 levels, which, collectively, may have restricted cell division, cell adhesion and neoangiogenesis. Taken together, these results showed that statin-resistant cells developed into smaller tumours than statin-sensitive cells. This may be reflective of the cancer restricting activity of statins in humans, as suggested from several retrospective studies with subjects undergoing statin therapy for several years.


Potential pathogenicity of Cladosporium tennuisimum, Phaeoisaria clematidis and Ramichloridium subulatum in a mouse model.
The potential pathogenicity of one isolate each of Cladosporium tennuisimum, Phaeoisaria clematidis and Ramichloridium subulatum for mice was investigated by intravenous, intraperitoneal and subcutaneous routes of inoculation with saline and mucin suspensions of the organisms. No mice died during the experimental period. Dark nodular lesions were formed in the liver, spleen, kidneys, intestine, stomach, omentum and diaphragm after inoculation through the intravenous and intraperitoneal routes. Gross lesions were produced in the lungs of one mouse inoculated intravenously with R. subulatum. Localised nodular lesions were formed in the subcutis following the subcutaneous route. Tissue response was characterised by granulomatous inflammatory reaction. Fungal elements were confined within the granulomata. Though the fungi showed limited pathogenic potential, they may represent a hazard under conditions of compromised host immunity.


Ethnopharmacology in the Upper Guadiana River area (Castile-La Mancha, Spain).
Determining traditional remedies for human pathologies is relevant, when compared with the standard materia medica of the pharmacopoeias and dietary supplement databases, because we can assess the species and uses that have been previously studied and target understudied species for further pharmacological investigation. The aim of this study was to systematically record and analyze medicinal uses of natural resources (Plantae, Animalia, Fungi and minerals), mostly local, in the territories adjoining the upper Guadiana River and its tributaries. We were particularly interested in recording resources and pathologies linked to wetland areas, especially in the National Parks Las Tablas de Daimiel and Cabañeros. Wetlands are interesting because they present a double face in relation with human health: Wetlands furnish hydration, safe water, nutrition, and medicinal resources; are places from which people derive their livelihood. However wetlands are also sites of exposure to pollution or toxicants, and infectious diseases; and sites of physical hazards. We wanted to identify procedures for preparation of medicinal formulae and routes of administration. We also intended to detect whether a geographical pattern exists or not in our records in relation to the use of local resources. We used semi-structured interviews with one-to-one informants or groups, from 1998 to 2018. Raw data were introduced in a Firebird database and analyzed. To identify ingredients and pathologies we consulted local floras and epidemiological literature. Finally, we compared documented pathologies, remedies and ingredients in the historical context of medicinal uses of natural resources in Castile-La Mancha and especially in Ciudad Real. 126 pathologies and 220 species furnishing ingredients have been recorded from the interviews. In total, 188 are plants and 20 animals. The most commonly used species include Malva sylvestris, Phlomis lychnitis, Genista tridentata and Thymus mastichina. Most records refer to flowers, or fruits, of locally available plant species, classified as Mediterranean, European or widespread that belong to the Lamiaceae, Compositae or Leguminosae. Ingredients which are collected in open shrublands, known as "garrigue", and dry grasslands furnish a relevant proportion of records while the imported ingredients remain marginal. The contribution of wetlands, riverine habitats and irrigated fields and gardens as a source of medicinal resources is 36% of the records. It is relatively high considering its limited presence in terms of total extension within the study area. The most frequently reported diseases are respiratory, gastrointestinal, dermatological and infectious or parasitic. Along the Guadiana River in the Ciudad Real province exists a wide and deep knowledge of traditional remedies for the treatment of common pathologies, based fundamentally on the use of local flora, fauna and mineral resources. The uses and ingredients documented are useful for further pharmacological investigation to improve health care for a wide range of pathologies.


Effects of the PPARgamma agonist pioglitazone on lipoprotein metabolism in patients with type 2 diabetes mellitus.
Elevated plasma levels of VLDL triglycerides (TGs) are characteristic of patients with type 2 diabetes mellitus (T2DM) and are associated with increased production rates (PRs) of VLDL TGs and apoB. Lipoprotein lipase-mediated (LPL-mediated) lipolysis of VLDL TGs may also be reduced in T2DM if the level of LPL is decreased and/or the level of plasma apoC-III, an inhibitor of LPL-mediated lipolysis, is increased. We studied the effects of pioglitazone (Pio), a PPARgamma agonist that improves insulin sensitivity, on lipoprotein metabolism in patients with T2DM. Pio treatment reduced TG levels by increasing the fractional clearance rate (FCR) of VLDL TGs from the circulation, without changing direct removal of VLDL particles. This indicated increased lipolysis of VLDL TGs during Pio treatment, a mechanism supported by our finding of increased plasma LPL mass and decreased levels of plasma apoC-III. Lower apoC-III levels were due to reduced apoC-III PRs. We saw no effects of Pio on the PR of either VLDL TG or VLDL apoB. Thus, Pio, a PPARgamma agonist, reduced VLDL TG levels by increasing LPL mass and inhibiting apoC-III PR. These 2 changes were associated with an increased FCR of VLDL TGs, almost certainly due to increased LPL-mediated lipolysis.


The Molecular and Physiological Effects of Protein-Derived Polyamines in the Intestine.
Consumption of a high-protein diet increases protein entry into the colon. Colonic microbiota can ferment proteins, which results in the production of protein fermentation end-products, like polyamines. This review describes the effects of polyamines on biochemical, cellular and physiological processes, with a focus on the colon. Polyamines (mainly spermine, spermidine, putrescine and cadaverine) are involved in the regulation of protein translation and gene transcription. In this, the spermidine-derived hypusination modification of EIF5A plays an important role. In addition, polyamines regulate metabolic functions. Through hypusination of EIF5A, polyamines also regulate translation of mitochondrial proteins, thereby increasing their expression. They can also induce mitophagy through various pathways, which helps to remove damaged organelles and improves cell survival. In addition, polyamines increase mitochondrial substrate oxidation by increasing mitochondrial Ca2+-levels. Putrescine can even serve as an energy source for enterocytes in the small intestine. By regulating the formation of the mitochondrial permeability transition pore, polyamines help maintain mitochondrial membrane integrity. However, their catabolism may also reduce metabolic functions by depleting intracellular acetyl-CoA levels, or through production of toxic by-products. Lastly, polyamines support gut physiology, by supporting barrier function, inducing gut maturation and increasing longevity. Polyamines thus play many roles, and their impact is strongly tissue- and dose-dependent. However, whether diet-derived increases in colonic luminal polyamine levels also impact intestinal physiology has not been resolved yet.


Late, not early mismatch responses to changes in frequency are reduced or deviant in children with dyslexia: an event-related potential study.
Developmental disorders of oral and written language have been linked to deficits in the processing of auditory information. However, findings have been inconsistent, both for behavioural and electrophysiological measures. In this study, we examined event-related potentials (ERPs) in 20 6- to 14-year-old children with developmental dyslexia and 20 age-matched controls, divided into younger (6-11 years, n = 10) and older (11-14 years, n = 10) age bands. We focused on early (mismatch negativity; MMN) and late (late discriminative negativity; LDN) conventional mismatch responses and associated measures derived from time-frequency analysis (inter-trial coherence and event-related spectral perturbation). Responses were elicited using an auditory oddball task, whereby a stream of 1000-Hz standards was interspersed with rare large (1,200 Hz) and small (1,030 Hz) frequency deviants. Conventional analyses revealed no significant differences between groups in the size of the MMN to either large or small frequency deviants. However, the younger age band of children with dyslexia showed an enhanced inter-trial coherence in the theta frequency band over the time window corresponding to the MMN to small deviants. By contrast, these same children showed a reduced-amplitude LDN for the small deviants relative to their age-matched controls, whilst the older children with dyslexia showed a shorter and less intense period of event-related desynchronization over this time window. Initial detection and discrimination of auditory frequency change appears normal or even enhanced in children with dyslexia. Rather, deficits in late-stage auditory processing appear to be a feature of this population.


Construction of bone marrow mesenchymal cells-derived engineered hepatic tissue and its therapeutic effect in rats with 90% subtotal hepatectomy.
Engineered hepatic tissue (EHT) is considered as a promising strategy for healing acute liver failure (ALF), therefore, in the present study we evaluated the therapeutic potential of the EHT which engaged with bone marrow mesenchymal cells (BMSCs) derived hepatocytes (BMSCs—Hepas) in ALF rats. After characterization of isolated BMSCs, we seeded passage 3 BMSCs which have being cultured in medium containing 20 ng/ml hepatocyte growth factor (HGF) and 10 ng/ml epidermal growth factor (EGF) for 14 days on three scaffolds individually in Transwell system, and then cultured for more than 3 days to construct three kinds of EHT named EHT1, EHT2, and EHT3. Based on morphology and urea production assays, we chose an optimal one and transplanted it into ALF rat with 90% subtotal hepatectomy and assessed its therapeutic potential by survival time, hepatic encephalopathy score (HES) and related liver function test. The remnant liver was acquired, sectioned and identified by con-focal scanning microscopy. The isolated cells possessed basic properties of BMSCs, when cultured in hepatogenic medium for 2 weeks, BMSCs would restore to the functional properties of primary rats' hepatocytes, expressing albumin (ALB) and alpha fetoprotein (AFP) simultaneously. Transplantation of EHT3 significantly prolonged the survival time, increased HES, and ameliorated the liver function. BMSC will be a newly cell source for the construction of EHT. Importantly, the EHT transplantation may be an effective strategy to treat ALF in clinic.


Subcutaneous infection with S. aureus in mice reveals association of resistance with influx of neutrophils and Th2 response.
Staphylococcus aureus is the leading cause of bacterial skin infection. Once it overcomes the epithelial barrier, it either remains locally controlled or spreads in the dermis causing soft tissue infection. These different courses depend not only on its virulence factors, but also on the immune response of the infected individual. The goal of this study was to identify host factors that influence different outcomes. We, therefore, established comparative analysis of subcutaneous footpad infection with S. aureus (SH1000) in different inbred mouse strains. We found that C57BL/6 mice are more susceptible than BALB/c and DBA/2 mice, reflected by significantly higher footpad swelling and bacterial load, as well as increased dissemination of bacteria into inguinal lymph nodes and kidneys. This susceptibility was associated with lower influx of polymorphonuclear leukocytes (PMNs), but higher secretion of CXCL-2. Remarkably, resistance correlated with S. aureus-specific Th2-cell response in BALB/c and DBA/2 mice, whereas susceptible C57BL/6 mice generated a Th1-cell response. As Th1 cells are able to induce release of CXCL-2, and as CXCL-2 is able to increase the survival of S. aureus within PMNs, interactions between PMNs and Th1 or Th2 cells need to be considered as important mechanisms of resistance in murine soft tissue infection with S. aureus.


Structures of FHOD1-Nesprin1/2 complexes reveal alternate binding modes for the FH3 domain of formins.
The nuclear position in eukaryotes is controlled by a nucleo-cytoskeletal network, critical in cell differentiation, division, and movement. Forces are transmitted through conserved Linker of Nucleoskeleton and Cytoskeleton (LINC) complexes that traverse the nuclear envelope and engage on either side of the membrane with diverse binding partners. Nesprin-2-giant (Nes2G), a LINC element in the outer nuclear membrane, connects to the actin directly as well as through FHOD1, a formin primarily involved in actin bundling. Here, we report the crystal structure of Nes2G bound to FHOD1 and show that the presumed G-binding domain of FHOD1 is rather a spectrin repeat (SR) binding enhancer for the neighboring FH3 domain. The structure reveals that SR binding by FHOD1 is likely not regulated by the diaphanous-autoregulatory domain helix of FHOD1. Finally, we establish that Nes1G also has one FHOD1 binding SR, indicating that these abundant, giant Nesprins have overlapping functions in actin-bundle recruitment for nuclear movement.


Leaf mechanisms for drought resistance in Zizyphus jujuba trees.
No information exists on the mechanisms developed at the level of leaf water relations by pear-jujube trees (Zizyphus jujuba Mill.) to confront drought. For this reason, the purpose of the present study was to analyse its leaf water relations in order to clarify the resistance mechanisms (avoidance and tolerance) developed in response to a water stress and during recovery. Field-grown 7-year-old pear-jujube trees (cv. Grande de Albatera) were subjected to three irrigation treatments. Control (T0) plants were drip irrigated (112% ETo) in order to guarantee non-limiting soil water conditions, T1 plants (deficit irrigation, 64% ETo) were drip irrigated according to the criteria used by the grower and T2 plants irrigated as T0 but subjected to water withholding for 36 days and a subsequent re-irrigation at the levels used in T0 for 14 days, during the summer of 2011. The results indicated that pear-jujube plants confront water stress by developing stress avoidance and stress tolerance mechanisms. From the beginning of deficit irrigation (T1) and water withholding (T2) to when maximum water stress levels were achieved, leaf turgor was maintained allowing substantial gas exchange levels and, consequently, good leaf productivity. This leaf turgor maintenance was mainly due to two simultaneous and complementary mechanisms. Leaf conductance and the duration of maximum stomatal opening in water stressed plants decreased in order to control water loss via transpiration, contributing to maintain leaf turgor (stress avoidance mechanisms). Also, the gradual recovery of g(l) observed after rewatering the plants can be considered as a mechanism for promoting leaf rehydration. In addition, from the beginning of the stress period, active osmotic adjustment operated, also contributing to the maintenance of leaf turgor (stress tolerance mechanism). The high RWC(a) levels and the possibility of increasing the accumulation of water in the apoplasm in response to water stress, supporting a steeper gradient in water potential between the leaf and the soil, which can be considered another drought tolerance characteristic in pear-jujube.


Impact of gastrojejunostomy diameter on long-term weight loss following laparoscopic gastric bypass: a follow-up study.
Stenosis of the gastrojejunostomy after laparoscopic Roux-en-Y gastric bypass is a common occurrence. We have previously presented data demonstrating that the use of a 25-mm circular stapler results in a decreased incidence of stenosis compared to the results of a 21-mm circular stapler (6.2 vs. 15.9%, P=0.03). One potential drawback of the larger-diameter stapler is the possibility for impaired long-term weight loss due to decreased restriction. We sought to determine the impact of circular stapler diameter on excess weight loss up to 5 years after surgery. Our initial technique for creating the gastrojejunostomy after laparoscopic gastric bypass involved the transgastric passage of a 21-mm circular stapler anvil (group 1). After a large initial experience, we switched to a 25-mm circular stapler (group 2). Follow-up data were entered prospectively into a computer database. Weight loss was recorded as percent of excess weight lost. Only patients with follow-up beyond 3 years postoperatively were eligible for inclusion. Group 1 consisted of 145 consecutive patients and group 2 consisted of 116 consecutive patients. There was no significant difference in weight loss between the groups at 3 (66.1 vs. 65.2%, P=0.76, n=134), 4 (66.4 vs. 58.6%, P=0.1, n=66), and 5 years after gastric bypass (62.7 vs. 57.5%, P=0.24, n=75). The use of a 25-mm circular stapler in laparoscopic gastric bypass operations instead of a 21-mm stapler does not result in significantly different long-term weight loss. The 25-mm stapler is preferred with our technique.


Facile template free synthesis of KLa(MoO4)2:Eu3+,Tb3+ microspheres and their multicolor tunable luminescence.
Trivalent rare-earth (RE(3+) = Eu(3+), Tb(3+)) ion activated KLa(MoO4)2 microspheres have been synthesized at 180 °C via a facile hydrothermal route without using any templates, surfactant, or other organic additives. X-ray diffraction (XRD), field emission-scanning electron microscopy (FE-SEM), photoluminescence (PL), and photoluminescent excitation spectra (PLE) were employed to characterize the samples. It is found that the pH value in the initial solution is responsible for crystal phase, shape determination and emission intensity of final products. The possible formation mechanism for products with uniform spheres has been presented. Furthermore, a systematic study on the photoluminescence of RE(3+) (RE(3+) = Eu(3+), Tb(3+)) doped KLa(MoO4)2 samples has been explored in order to obtain the multicolor tunable emission by varying the Tb(3+)/Eu(3+) ratio. The tunable luminescence may be potentially applied in fields such as solid state lighting and field emission displays.


Interplay between the membrane-associated UhpB and UhpC regulatory proteins.
Expression of the Escherichia coli uhpT gene, encoding the sugar phosphate transport protein, is induced by extracellular glucose-6-phosphate and requires the function of the uhpABC regulatory genes. The UhpA and UhpB proteins are related to the response-regulator and sensor-kinase proteins of two-component regulatory systems, whereas the UhpC protein is related to UhpT and homologous transport proteins. To investigate the role of segments of the membrane-associated UhpB and UhpC regulatory proteins, a series of mutations were constructed in vitro by insertion of a 12- or 24-bp oligonucleotide linker at 44 sites within the uhpABCT locus. The effect of these mutations on regulation of a uhpT-lacZ transcriptional reporter was assayed with the mutated uhp alleles in single copy on the chromosome. All but one of the insertions in uhpA or uhpT were inactive for transcription activation or transport, respectively. In contrast, about half of the insertions in uhpB and uhpC retained Uhp expression, and insertions at four sites in uhpB and at one site in uhpC conferred high-level constitutive expression. The constitutive mutants in UhpB resulted from insertions in the nonpolar amino-terminal half of the protein, and all insertions in that half of UhpB affected Uhp expression in some manner, which suggests that the transmembrane segments of UhpB might negatively regulate the kinase activity of the carboxyl portion. The constitutive behavior of all but one of these uhpB alleles was dependent on the presence of active forms of both UhpA and UhpC, which suggests that UhpB and UhpC act jointly as a complex in the signaling process.


CARIOSTATIC EFFECT OF PHOSPHATES.
The caries-preventive effect of phosphate additives in cariogenic diets fed to white rats has been further demonstrated. Diammonium phosphate, beta-glycerol phosphate, and sodium phytate again were shown to be caries-inhibiting, as was a 1,6-fructose diphosphate. The caries-inhibiting action of organic phosphates may coincide with a caries-protective factor presumed lost in the refining of sugar and in the processing of certain cereal foods.


Developing robust arsenic awareness prediction models using machine learning algorithms.
Arsenic awareness plays a vital role in ensuring the sustainability of arsenic mitigation technologies. Thus far, however, few studies have dealt with the sustainability of such technologies and its associated socioeconomic dimensions. As a result, arsenic awareness prediction has not yet been fully conceptualized. Accordingly, this study evaluated arsenic awareness among arsenic-affected communities in rural India, using a structured questionnaire to record socioeconomic, demographic, and other sociobehavioral factors with an eye to assessing their association with and influence on arsenic awareness. First a logistic regression model was applied and its results compared with those produced by six state-of-the-art machine-learning algorithms (Support Vector Machine [SVM], Kernel-SVM, Decision Tree [DT], k-Nearest Neighbor [k-NN], Naïve Bayes [NB], and Random Forests [RF]) as measured by their accuracy at predicting arsenic awareness. Most (63%) of the surveyed population was found to be arsenic-aware. Significant arsenic awareness predictors were divided into three types: (1) socioeconomic factors: caste, education level, and occupation; (2) water and sanitation behavior factors: number of family members involved in water collection, distance traveled and time spent for water collection, places for defecation, and materials used for handwashing after defecation; and (3) social capital and trust factors: presence of anganwadi and people's trust in other community members, NGOs, and private agencies. Moreover, individuals' having higher social network positively contributed to arsenic awareness in the communities. Results indicated that both the SVM and the RF algorithms outperformed at overall prediction of arsenic awareness-a nonlinear classification problem. Lower-caste, less educated, and unemployed members of the population were found to be the most vulnerable, requiring immediate arsenic mitigation. To this end, local social institutions and NGOs could play a crucial role in arsenic awareness and outreach programs. Use of SVM or RF or a combination of the two, together with use of a larger sample size, could enhance the accuracy of arsenic awareness prediction.


Nutrigenomics, individualism and public health.
Issues arising in connection with genes and nutrition policy include both nutrigenomics and nutrigenetics. Nutrigenomics considers the relationship between specifc nutrients or diet and gene expression and, it is envisaged, will facilitate prevention of diet-related common diseases. Nutrigenetics is concerned with the effects of individual genetic variation (single nucleotide polymorphisms) on response to diet, and in the longer term may lead to personalised dietary recommendations. It is important also to consider the surrounding context of other issues such as novel and functional foods in so far as they are related to genetic modification. Ethical issues fall into a number of categories: (1) why nutrigenomics? Will it have important public health benefits? (2) questions about research, e.g. concerning the acquisition of information about individual genetic variation; (3) questions about who has access to this information, and its possible misuse; (4) the applications of this information in terms of public health policy, and the negotiation of the potential tension between the interests of the individual in relation to, for example, prevention of conditions such as obesity and allergy; (5) the appropriate ethical approach to the issues, e.g. the moral difference, if any, between therapy and enhancement in relation to individualised diets; whether the 'technological fix' is always appropriate, especially in the wider context of the purported lack of public confidence in science, which has special resonance in the sphere of nutrition.


Gastrointestinal anomalies associated with esophageal atresia or tracheoesophageal fistula.
A retrospective analysis of 150 consecutive patients with esophageal atresia and/or tracheoesophageal fistula was undertaken. The incidence and variety of gastrointestinal (GI) anomalies, the method of management, and the cause of death were critically reviewed to outline preferred methods of management. Of these patients, 87 were male and 63 female. There were 62 GI anomalies in 40 patients. The most common GI anomalies included anorectal anomalies in 17 patients, malrotation of the midgut in 13 patients, duodenal atresia or stenosis in ten patients, and annular pancreas and ectopic pancreatic tissue in three patients each. Six patients had the combination of tracheoesophageal fistula, duodenal obstruction, and anorectal anomaly. Of the 150 patients, 23 have died, with 15 deaths occurring in those with associated GI anomalies. The GI anomalies contributed directly to the deaths of five of these 15 patients. Delay in diagnosis and, on occasion, improper initial management resulted in increased morbidity and mortality.


High-resolution crystal structures of Caldicellulosiruptor strain Rt8B.4 carbohydrate-binding module CBM27-1 and its complex with mannohexaose.
Carbohydrate-binding modules (CBMs) are the most common non-catalytic modules associated with enzymes active in plant cell-wall hydrolysis. Despite the large number of putative CBMs being identified by amino acid sequence alignments, only few representatives have been experimentally shown to have a carbohydrate-binding function. Caldicellulosiruptor strain Rt8B.4 Man26 is a thermostable modular glycoside hydrolase beta-mannanase which contains two non-catalytic modules in tandem at its N terminus. These modules were recently shown to function primarily as beta-mannan-binding modules and have accordingly been classified as members of a novel family of CBMs, family 27. The N-terminal CBM27 (CsCBM27-1) of Man26 from Caldicellulosiruptor Rt8B.4 displays high-binding affinity towards mannohexaose with a Ka of 1 x 10(7) M(-1). Accordingly, the high-resolution crystal structures of CsCBM27-1 native and its mannohexaose complex were solved at 1.55 angstroms and 1.06 angstoms resolution, respectively. In the crystal, CsCBM27-1 shows the typical beta-sandwich jellyroll fold observed in other CBMs with a single metal ion bound, which was identified as calcium. The crystal structures reveal that the overall fold of CsCBM27-1 remains virtually unchanged upon sugar binding and that binding is mediated by three solvent-exposed tryptophan residues and few direct hydrogen bonds. Based on binding affinity and thermal unfolding experiments this structural calcium is shown to play a role in the thermal stability of CsCBM27-1 at high temperatures. The higher binding affinity of CsCBM27-1 to mannooligosaccharides when compared to other members of CBM family 27 might be explained by the different orientation of the residues forming the "aromatic platform" and by differences in the length of loops. Finally, evidence is presented, on the basis of fold similarities and the retention of the position of conserved motifs and a calcium ion, for the consolidation of related CBM families into a superfamily of CBMs.


Complete genome sequence of Desulfobulbus propionicus type strain (1pr3).
Desulfobulbus propionicus Widdel 1981 is the type species of the genus Desulfobulbus, which belongs to the family Desulfobulbaceae. The species is of interest because of its great implication in the sulfur cycle in aquatic sediments, its large substrate spectrum and a broad versatility in using various fermentation pathways. The species was the first example of a pure culture known to disproportionate elemental sulfur to sulfate and sulfide. This is the first completed genome sequence of a member of the genus Desulfobulbus and the third published genome sequence from a member of the family Desulfobulbaceae. The 3,851,869 bp long genome with its 3,351 protein-coding and 57 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.


Chromatin control and cancer-drug discovery: realizing the promise.
Recent years have seen major advances in elucidating the complexity of chromatin and its role as an epigenetic regulator of gene expression in eukaryotes. We now have a basic understanding of chromatin control and the enzymatic modifications that impart diverse regulatory cues to the functional activity of the genome. Most importantly, although research into chromatin has uncovered fascinating insights into the control of gene expression, it has also generated a large body of information that is being harnessed to develop new therapeutic modalities for treating cancer. Here, we discuss recent advances that support the contention that future generations of chromatin-modulating drugs will provide a significant group of new, mechanism-based therapeutics for cancer.


A Review of Questionnaire Measures for Assessing the Social Climate in Prisons and Forensic Psychiatric Hospitals.
Significant time and resources are devoted to the monitoring of social climate in secure settings. However, if these efforts are to help (rather than hinder) attempts to improve the functioning of such units, the monitoring of social climate must be based on sound psychometric methods. The aim of this review was to determine what questionnaires exist to measure the social climate in secure settings and what evidence exists regarding their psychometric properties. Twelve questionnaire-based measures of social climate were identified. The Essen Climate Evaluation Schema has received the most consistent empirical support, but this questionnaire does not provide as much of an in-depth, detailed insight into social climate as that provided by other social climate questionnaires. Although more extensive measures of climate exist, they have not yet received sufficient validation to justify their routine use in practice. Nevertheless, there is growing evidence that some questionnaire-based measures can provide a reliable and valid assessment of the social climate in secure settings, which has important clinical and theoretical implications.


Incidence of Anaphylaxis to Sugammadex in a Single-Center Cohort of 19,821 Patients.
Sugammadex is a modified cyclodextrin that is being increasingly used in anesthetic practice worldwide for the reversal of the aminosteroid neuromuscular blockers rocuronium and vecuronium. Its safety profile, however, is incompletely understood. One such aspect is the incidence of anaphylactic reactions that occur after its administration. While several case reports exist in the literature, there is a paucity of information on the actual incidence of anaphylactic reactions. A single-center retrospective chart review identified patients who experienced anaphylaxis to sugammadex in the institutional electronic medical record system. These charts were then reviewed to determine whether the etiology of anaphylaxis was sugammadex administration. Two patients experienced anaphylaxis to sugammadex, which occurred in a single institution cohort of 19,821 patients who received 23,446 total doses. This rate is markedly lower than the 1/300 that the manufacturer's package insert states and also lower than the 1/2500 that the only other large cohort study performed has reported. The incidence of anaphylaxis to sugammadex in this cohort of patients was 2 of 19,821 patients, who received a total of 23,446 doses.


The Clinical Efficacy of Mometasone Furoate in Multi-Lamellar Emulsion for Eczema: A Double-blinded Crossover Study.
Topical application of corticosteroids also has an influence on skin barrier impairment. Physiological lipid mixtures, such as multi-lamellar emulsion (MLE) containing a natural lipid component leads to effective recovery of the barrier function. The purpose of this study was to conduct an evaluation of the therapeutic efficacy and skin barrier protection of topical mometasone furoate in MLE. A multi-center randomized, double-blind, controlled study was performed to assess the efficacy and safety of mometasone furoate cream in MLE for Korean patients with eczema. The study group included 175 patients with eczema, who applied either mometasone furoate in MLE cream or methylprednisolone aceponate cream for 2 weeks. Treatment efficacy was evaluated using the physician's global assessment of clinical response (PGA), trans-epidermal water loss (TEWL), and visual analogue scale (VAS) for pruritus. Patients were evaluated using these indices at days 4, 8, and 15. Comparison of PGA score, TEWL, and VAS score at baseline with those at days 4, 8, and 15 of treatment showed a significant improvement in both groups. Patients who applied mometasone furoate in MLE (74.8%) showed better results (p<0.05) than those who applied methylprednisolone aceponate (47.8%). The TEWL improvement ratio was higher in the mometasone furoate in MLE group than that in the methylprednisolone aceponate group, and VAS improvement was also better in the mometasone furoate in MLE group. Mometasone furoate in MLE has a better therapeutic efficacy as well as less skin barrier impairment than methylprednisolone aceponate.


A missense mutation in the SERPINH1 gene in Dachshunds with osteogenesis imperfecta.
Osteogenesis imperfecta (OI) is a hereditary disease occurring in humans and dogs. It is characterized by extremely fragile bones and teeth. Most human and some canine OI cases are caused by mutations in the COL1A1 and COL1A2 genes encoding the subunits of collagen I. Recently, mutations in the CRTAP and LEPRE1 genes were found to cause some rare forms of human OI. Many OI cases exist where the causative mutation has not yet been found. We investigated Dachshunds with an autosomal recessive form of OI. Genotyping only five affected dogs on the 50 k canine SNP chip allowed us to localize the causative mutation to a 5.82 Mb interval on chromosome 21 by homozygosity mapping. Haplotype analysis of five additional carriers narrowed the interval further down to 4.74 Mb. The SERPINH1 gene is located within this interval and encodes an essential chaperone involved in the correct folding of the collagen triple helix. Therefore, we considered SERPINH1 a positional and functional candidate gene and performed mutation analysis in affected and control Dachshunds. A missense mutation (c.977C>T, p.L326P) located in an evolutionary conserved domain was perfectly associated with the OI phenotype. We thus have identified a candidate causative mutation for OI in Dachshunds and identified a fifth OI gene.


DNA damage and tumor surveillance: one trigger for two pathways.
Biological evolution ensures that several backup mechanisms always exist to prevent the failure of cellular processes of critical importance for life. This notion applies to processes aimed at preventing cancer development. Recent research suggests that the DNA damage response, activated in one of the earliest cellular responses to transformation, may elicit two independent tumor surveillance mechanisms. The first, and most well known, is activation of the proapoptotic molecule p53 and subsequent cellular suicide. The second, reported in a recent study, is induction of the expression of ligands for the activating immune receptor NKG2D. That DNA damage induces two independent tumor surveillance responses demonstrates how one tumor surveillance mechanism may be assisted by another to secure efficient protection against early tumorigenesis. The results also support the immunosurveillance theory of cancer and suggest that it may operate at very early stages of tumorigenesis.


Effects of O-methylated metabolites of quercetin on oxidative stress, thermotolerance, lifespan and bioavailability on Caenorhabditis elegans.
Quercetin is a major flavonoid in the human diet and the most commonly used in studies of biological activity. Most of the knowledge about its biological effects has originated from in vitro studies while in vivo data are scarce. Quercetin mostly occurs in foodstuffs as glycosides that are deglycosylated during absorption and further submitted to different conjugation reactions. Methylation to isorhamnetin (quercetin 3'-O-methylether) or tamarixetin (quercetin 4'-O-methylether) seems to be an important conjugation process in quercetin metabolism. In this work, the effects of quercetin and its 3'- and 4'-O-methylated metabolites on the phenotypic characteristics, stress oxidative resistance, thermotolerance and lifespan of the model organism Caenorhabditis elegans have been assessed. The three assayed flavonols significantly prolonged the lifespan of this nematode with an increase from 11% to 16% in the mean lifespan with respect to controls. However, only quercetin significantly increased the reproductive capacity of the worm and enlarged the body size. Exposure to the assayed flavonols also increased significantly the resistance against thermal and juglone-induced oxidative stress, although differences were found depending on the stage of development of the worm. Thus, quercetin offered greater protection when thermal stress was applied in the 1st day of adulthood, whereas tamarixetin was more efficient in worms submitted to stress in the 6th day of adulthood. Similarly, significantly greater protection was provided by quercetin than by its methylated derivatives at the 1st day of adulthood, whilst quercetin and isorhamnetin were equally efficient when the oxidative stress was induced in the 6th of day of adulthood. Further evidence of antioxidant protection was obtained checking the oxidation status of proteins by the OxyBlot™ detection kit. Analyses by HPLC-DAD-ESI/MS confirmed that the three flavonols were taken up by C. elegans leading to the formation of some glycosylated, sulfated and methylated metabolites, and that demethylation of these latter to quercetin was also produced. Quantification of the levels of quercetin, isorhamnetin and tamarixetin, as well as their detected metabolites indicated a greater uptake of quercetin than its methylated derivatives by the nematode.


Strategies for Enriching Variant Coverage in Candidate Disease Loci on a Multiethnic Genotyping Array.
Investigating genetic architecture of complex traits in ancestrally diverse populations is imperative to understand the etiology of disease. However, the current paucity of genetic research in people of African and Latin American ancestry, Hispanic and indigenous peoples in the United States is likely to exacerbate existing health disparities for many common diseases. The Population Architecture using Genomics and Epidemiology, Phase II (PAGE II), Study was initiated in 2013 by the National Human Genome Research Institute to expand our understanding of complex trait loci in ethnically diverse and well characterized study populations. To meet this goal, the Multi-Ethnic Genotyping Array (MEGA) was designed to substantially improve fine-mapping and functional discovery by increasing variant coverage across multiple ethnicities at known loci for metabolic, cardiovascular, renal, inflammatory, anthropometric, and a variety of lifestyle traits. Studying the frequency distribution of clinically relevant mutations, putative risk alleles, and known functional variants across multiple populations will provide important insight into the genetic architecture of complex diseases and facilitate the discovery of novel, sometimes population-specific, disease associations. DNA samples from 51,650 self-identified African ancestry (17,328), Hispanic/Latino (22,379), Asian/Pacific Islander (8,640), and American Indian (653) and an additional 2,650 participants of either South Asian or European ancestry, and other reference panels have been genotyped on MEGA by PAGE II. MEGA was designed as a new resource for studying ancestrally diverse populations. Here, we describe the methodology for selecting trait-specific content for use in multi-ethnic populations and how enriching MEGA for this content may contribute to deeper biological understanding of the genetic etiology of complex disease.


The Agrobacterium tumefaciens VirB7 lipoprotein is required for stabilization of VirB proteins during assembly of the T-complex transport apparatus.
The Agrobacterium tumefaciens virB7 gene product is a lipoprotein whose function is required for the transmission of oncogenic T-DNA to susceptible plant cells. Three lines of study provided evidence that VirB7 interacts with and stabilizes other VirB proteins during the assembly of the putative T-complex transport apparatus. First, a precise deletion of virB7 from the pTiA6NC plasmid of wild-type strain A348 was correlated with significant reductions in the steady-state levels of several VirB proteins, including VirB4, VirB9, VirB10, and VirB11; trans expression of virB7 in the delta virB7 mutant partially restored the levels of these proteins, and trans coexpression of virB7 and virB8 fully restored the levels of these proteins to wild-type levels. Second, modulation of VirB7 levels resulted in corresponding changes in the levels of other VirB proteins in the following cell types: (i) a delta virB7 mutant expressing virB7 and virB8 from isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible Plac and other virB genes from acetosyringone (AS)-inducible PvirB; (ii) a delta virB operon mutant expressing virB7 and virB8 from Plac and virB9, virB10, and virB11 from PvirB; and (iii) a delta virB operon mutant expressing virB7 from IPTG-inducible Pklac and virB9 from an AS-inducible PvirB. Third, the synthesis of a VirB7::PhoA fusion protein in strain A348 was correlated with a significant reduction in the steady-state levels of VirB4, VirB5, and VirB7 through VirB11; these cells also exhibited a severely attenuated virulence phenotype, indicating that synthesis of the fusion protein perturbs the assembly of VirB proteins into a stabilized protein complex required for T-complex transport. Extracts of AS-induced cells electrophoresed under nonreducing conditions possessed undetectable levels of the 32-kDa VirB9 and 4.5-kDa VirB7 monomers and instead possessed a 36-kDa complex that cross-reacted with both VirB7 and VirB9 antisera and accumulated as a function of virB7 expression. Our results are consistent with a model in which VirB7 stabilizes VirB9 by formation of a covalent intermolecular cross-link; in turn, the VirB7-VirB9 heterodimer promotes the assembly of a functional T-complex transport machinery.


Long-term symptomatic outcomes after Collis gastroplasty with fundoplication.
Collis gastroplasty with fundoplication is an accepted treatment for gastroesophageal reflux disease (GERD) complicated by short esophagus. The procedure can be done either via left thoracotomy or using minimally invasive laparoscopic techniques. Few centers have reported long-term follow-up for patients undergoing a Collis gastroplasty using both the open and minimal access techniques. Retrospective review of prospectively collected data at Creighton University was done to identify patients who underwent Collis gastroplasty with fundoplication for GERD. After approval from the institutional review board, the patients were contacted and administered a questionnaire regarding symptoms and satisfaction. Data were entered in a dataset and analyzed from the patient's perspective. Eighty-five patients underwent a Collis gastroplasty procedure over a period of 13 years. Forty-eight percent (41 cases) were performed laparoscopically, and a transthoracic open repair was performed in the rest. Long-term data (more than 9 months) was available on 52 patients. Surgery resulted in complete resolution of heartburn, chest pain, regurgitation, and dysphagia in 52, 22, 54, and 29% of patients, respectively. More than 75% of the patients were satisfied with the outcome of surgery, and more than 85% would recommend the procedure to another patient. Collis gastroplasty with fundoplication results in good long-term patient satisfaction and symptom control.


Association between vitamin D and age-related macular degeneration in the Third National Health and Nutrition Examination Survey, 1988 through 1994.
To evaluate the associations between levels of vitamin D (25-hydroxyvitamin D) in serum and prevalent age-related macular degeneration (AMD). Cross-sectional associations of serum vitamin D and early and advanced AMD, assessed from nonmydriatic fundus photographs, were evaluated in the third National Health and Nutrition Examination Survey, a multistage nationally representative probability sample of noninstitutionalized individuals (N = 7752; 11% with AMD). Levels of serum vitamin D were inversely associated with early AMD but not advanced AMD. The odds ratio (OR) and 95% confidence interval (CI) for early AMD among participants in the highest vs lowest quintile of serum vitamin D was 0.64 (95% CI, 0.5-0.8; P trend <.001). Exploratory analyses were conducted to evaluate associations with important food and supplemental sources of vitamin D. Milk intake was inversely associated with early AMD (OR, 0.75; 95% CI, 0.6-0.9). Fish intake was inversely associated with advanced AMD (OR, 0.41; 95% CI, 0.2-0.9). Consistent use vs nonuse of vitamin D from supplements was inversely associated with early AMD only in individuals who did not consume milk daily (early AMD: OR, 0.67; 95% CI, 0.5-0.9). This study provides evidence that vitamin D may protect against AMD. Additional studies are needed to confirm these findings.


Relating the effects of protein type and content in increased-protein cheese pies to consumers' perception of satiating capacity.
Since proteins have been shown to have the highest satiation-inducing effects of all the macronutrients, increasing the protein level is one of the main strategies for designing foods with enhanced satiating capacity. However, few studies analyze the effect that protein addition has on the texture and flavor characteristics of the target food item to relate it to the expected satiating capacity it elicits. The present work studied cheese pies with three levels of soy and whey proteins. Since the protein level altered the rheological behavior of the batters before baking and the texture of the baked pies, the feasibility of adding several protein levels for obtaining a range of final products was investigated. A check-all-that-apply questionnaire containing 32 sensory and non-sensory characteristics of the samples was given to consumers (n = 131) who also scored the perceived samples' satiating capacity. The results showed that the type and content of protein contributed distinctive sensory characteristics to the samples that could be related to their satiating capacity perception. Harder and drier samples (high protein levels) were perceived as more satiating with less perceptible sweet and milky cheese pie characteristic flavors. Soy contributed an off-flavour. These results will contribute to a better understanding of the interrelation of all these factors, aiding the development of highly palatable solid foods with enhanced satiating capacities.


Lower limb length and offset in total hip arthroplasty.
Restoration of normal hip biomechanics is a key goal of total hip arthroplasty (THA) and favorably affects functional recovery. Furthermore, a major concern for both the surgeon and the patient is preservation or restoration of limb length equality, which must be achieved without compromising the stability of the prosthesis. Here, definitions are given for anatomic and functional limb length discrepancies and for femoral and hip offset, determined taking anteversion into account. Data on the influence of operated-limb length and offset on patient satisfaction, hip function, and prosthesis survival after THA are reviewed. Errors may adversely impact function, quality of life, and prosthetic survival and may also generate conflicts between the surgeon and patient. Surgeons rely on two- or three-dimensional preoperative templating and on intraoperative landmarks to manage offset and length. Accuracy can be improved by using computer-assisted planning or surgery and the more recently introduced EOS imaging system. The prosthetic's armamentarium now includes varus-aligned and lateralized implants, as well as implants with modular or custom-made necks, which allow restoration of the normal hip geometry, most notably in patients with coxa vara or coxa valga. Femoral anteversion must also receive careful attention. The most common errors are limb lengthening and a decrease in hip offset. When symptoms are caused by an error in length and/or offset, revision arthroplasty may deserve consideration.


Unraveling the time cross correlations of an emitter switching between two states with the same fluorescence intensity.
The autocorrelation function of the fluorescence intensity of a nanoemitter is measured with the standard Hanbury-Brown and Twiss setup. Time-tagging of the photodetection events during all the experiment has opened new possibilities in terms of post-selection techniques that enable to go beyond the blinking and antibunching characterization. Here, we first present a new method developed to investigate in detail the antibunching of a fluorophore switching between two emitting states. Even if they exhibit the same fluorescence intensity, their respective amount of antibunching can be measured using the gap between their respective decay rates. The method is then applied to a nanoemitter consisting in a colloidal quantum dot coupled to a plasmonic resonator. The relative quantum efficiency of the charged and neutral biexcitons are determined.