Protective effects of Ginkgo biloba extract (EGB 761) on astrocytes of rat hippocampus after exposure with scopolamine.
The regular extract of Ginkgo biloba has been shown to possess neuroprotective properties in disorders like hypoxia, ischemia, seizure activity and peripheral nerve damage. Also, G. biloba has received attention as a potential cognitive enhancer for the treatment of Alzheimer's disease, but there is not any documentation about the effect of an extract of G. biloba on astrocytes. Therefore, the aim of this study was examined the effects of G. biloba extract on the rat's hippocampal astrocytes after scopolamine based amnesia. In this study, 36 adult male Wistar rats were used. Rats were randomly distributed into control, sham, protective and treatment groups. The rats in the sham group only received scopolamine hydrobromide (3 mg/kg) intraperitoneally. The rats in the protective and treatment groups received G. biloba extract (40, 80 mg/kg) for 7 days intraperitoneally before and after scopolamine injection. Forty eight hours after the last injection, the brains of the rats were withdrawn and fixed with paraformaldehide, and then after histological processing, the slices were stained with phosphotungstic acid-haematoxylin for astrocytes. Data were analyzed by the analysis of variance (ANOVA) post hoc Tukey test; P<0.05 was considered significant. Results showed that scopolamine can reduce the number of astrocytes in all areas of hippocampal formation compared with the control. However, G. biloba extract can compensate for the reduction in the number of astrocytes in the hippocampus before or after the encounter with scopolamine. We concluded that a pretreatment and treatment injection of G. biloba extract can have a protective effect for astrocytes in all areas of hippocampal formation.


Lipidomics of equine amniotic fluid: Identification of amphiphilic (O-acyl)-ω-hydroxy-fatty acids.
Amniotic fluid is essential for the growth and maturation of the fetus prior to parturition. While our knowledge of human amniotic fluid is extensive, current data for equine amniotic fluid is limited. We therefore undertook a detailed lipidomics analysis of equine amniotic fluid. Using a non-targeted high-resolution mass spectrometric lipidomics analysis of equine amniotic fluid, we were able to characterize a diverse array of individual lipids. This non-biased analytical approach detected, for the first time, the presence of (O-acyl)-ω-hydroxy-fatty acids (OAHFA) with up to 52 carbon chain lengths in amniotic fluid. The identities of these lipid amphiphiles were validated both by high-resolution mass spectrometry and by tandem mass spectrometry (<2 ppm mass error) which identified the fatty acid and hydroxy-fatty acid components of individual OAHFAs. The only previous identification of OAHFAs has been in sperm and meibomian glands, and their sebaceous secretions, suggesting that these lipids may have unique functional roles in highly specialized compartments. The amphiphilic and surfactant properties of these unique lipids could provide an interface between amniotic lipids and fetal skin and/or lungs. The potential roles of OAHFAs as well as their source in amniotic fluid remain to be explored based upon these novel lipidomics findings but our study of the developmental time course of amniotic OAHFAs suggest that they may act as lubricants in delivery and/or a role in the development of fetal lung function around parturition.


LL-37 directs macrophage differentiation toward macrophages with a proinflammatory signature.
The human cathelicidin LL-37 has broad-spectrum antimicrobial activity. It also participates at the interface of innate and adaptive immunity by chemoattracting immune effector cells, modulating the production of a variety of inflammatory mediators by different cell types, and regulating the differentiation of monocytes into dendritic cells. In this study, we investigated the effects of LL-37 on the differentiation of human monocytes into anti-inflammatory macrophages (MPhi-2; driven by M-CSF) versus proinflammatory macrophages (MPhi-1; driven by GM-CSF) as well as on fully differentiated MPhi-1 and MPhi-2. Results revealed that monocytes cultured with M-CSF in the presence of LL-37 resulted in macrophages displaying a proinflammatory signature, namely, low expression of CD163 and little IL-10 and profound IL-12p40 production on LPS stimulation. The effects of LL-37 on M-CSF-driven macrophage differentiation were dose- and time-dependent with maximal effects observed at 10 microg/ml when the peptide was present from the start of the cultures. The peptide enhanced the GM-CSF-driven macrophage differentiation. Exposure of fully differentiated MPhi-2 to LL-37 for 6 d resulted in macrophages that produced less IL-10 and more IL-12p40 on LPS stimulation than control MPhi-2. In contrast, LL-37 had no effect on fully differentiated MPhi-1. Peptide mapping using a set of 16 overlapping 22-mer peptides covering the complete LL-37 sequence revealed that the C-terminal portion of LL-37 is responsible for directing macrophage differentiation. Our results furthermore indicate that the effects of LL-37 on macrophage differentiation required internalization of the peptide. Together, we conclude that LL-37 directs macrophage differentiation toward macrophages with a proinflammatory signature.


Escherichia coli and Serratia fonticola ESBLs as a potential source of antibiotics resistance dissemination in the Tricity water reservoirs.
Despite the fact that cephalosporins are rarely used in medical or veterinary treatment, the presence of Enterobacterales strains resistant to this group of anti-bacterial drugs (ESBL) is an important issue that requires attention. Between 2019 and 2021, 14 retention reservoirs, 12 streams, 3 rivers and 1 lake situated in the Tricity area (in northern Poland) were sampled for the presence of ESBL strains. Out of 40 water samples, characteristic growth (E. coli and the KESC group) on Chromagar ESBL plates was observed for 33 samples. The average number of ESBL E. coli was 42±132 CFU/100 ml, while the KESC group was 73±147 CFU/100 ml. Out of 33 positive samples, 57 ESBL Enterobacterales strains were isolated, of which the most abundant species were E. coli (13 isolates) and S. fonticola (23 isolates). The E. coli ESBL isolates not only showed resistance to third generation cephalosporins but also to antibiotics from other groups, such as fluoroquinolones, aminoglycosides and sulfonamides. The S. fonticola ESBL isolates were also found to be mainly resistant to the third generation cephalosporins, with the exception of 5 imipenem and 2 ertapenem-resistant strains. These strains presented highly diverse fingerprinting profiles, as well as significant differences in phenotypic traits helpful for survival in the environment, such as biofilm formation and motility. Moreover, biofilm formation and the swimming ability were species and temperature dependent. We confirmed the presence of highly diverse ESBL strains with multiple drug resistance patterns in the Tricity water reservoirs. This could possibly pose a threat to human health and create a suitable ground for acquiring antibiotics resistance in the natural environment.


Prevalence of hearing loss and use of hearing aids among children and adolescents in Germany: a systematic review.
Current data suggest that approximately 466 million people (5.0%) of the world's population have disabling hearing loss, therefrom, 34 million children, impacting their quality of life. To provide estimates on the prevalence of hearing loss on a national level, we reviewed the epidemiological literature addressing hearing loss in children and adolescents living in Germany as an example for a Western country. We searched Medline, Web of Science, Cochrane Library, ScienceDirect and LIVIVO to identify published data. Furthermore, we manually searched websites of relevant institutions and journals not listed in electronically and searched for ongoing studies and/or not yet published data in clinicaltrials.gov . Study selection, data extraction, and methodological assessment were carried out by two reviewers. In total, 11 reports provided data with sample sizes ranging from 310 up to more than 14 million children and adolescents. Prevalence data were collected by interviews (self-assessments), using pure-tone audiometry or the international classification of diseases (ICD-10) coding and ranged from 0.1 to 128 per 1000 children. Although the estimate of the prevalence of hearing loss goes down, when the threshold was raised, generating a comprehensive and coherent set of estimates proved challenging owing to clinical heterogeneity including variation in age, the study setting, the definition of hearing loss and the assessment method. Moreover, representativeness (external validity) was often impaired owing to estimates lacking currentness (i.e., referring to former West Germany) or selected (patient) data and may not be typical for a more general population. In conclusions, this work raises public awareness of the high prevalence of hearing loss, highlights issues associated with epidemiological research and is of great importance for researcher and those who use epidemiological data to inform clinical and political decision making.


Severe disease in patients with rheumatoid arthritis carrying a mutation in the Mediterranean fever gene.
Pyrin is a newly recognised intracellular regulator of inflammation, and mutations in MEFV, the gene encoding pyrin, are the cause of familial Mediterranean fever. To determine if known mutations of MEFV are associated with rheumatoid arthritis (RA) morbidity or can modify RA severity. The frequency of the three most common MEFV mutations: M694V, V726A, and E148Q, was determined in 98 Israeli patients with RA (74 women, 24 men) and compared with that in 100 healthy subjects matched for origin. RA severity was determined using a new clinical score of 126 grades. The median severity score of mutation carrier and non-carrier groups was compared after confounding measures were eliminated by logistic regression. 17/98 (17%) patients with RA (all women) were heterozygous for common MEFV mutations, predominantly E148Q (12 patients), and one patient was homozygous for the V726A mutation. The overall mutation rate was comparable between patients with RA and healthy subjects. Patients carrying a mutation had a higher median severity score than the non-carrier group (42 v 29, p = 0.0005). The logistic regression model assigned a 15-fold odds ratio for severe RA in carriers, after adjusting for sex, presence of rheumatoid factor, age at onset, and disease duration (n = 97, p = 0.01, 95% CI 1.74 to 128). MEFV, and particularly the E148Q mutation, is an independent modifier of the clinical manifestations of RA. This is the second Th1-type autoimmune disease in which MEFV mutations have been shown to aggravate the clinical status.


Use of free vascularized bone grafts in the treatment of bone tumors.
A five-year experience with ten cases of primary resection of aggressive bone lesions treated by immediate reconstruction with autologous fibula demonstrates the advantages of vascularized, compared with free bone, graft. The procedure was performed for locally aggressive bone tumors in seven cases, and for Type III congenital pseudarthroses of the tibia in three cases. In large intercalary bone grafts, the vascularized fibula actively participates in the healing process, rather than serving merely as a framework for inductive creeping substitution, as is the case in conventional bone grafts. With the exception of one case, the grafts united and progressively hypertrophied, and there was no tumor recurrence. For defects less than 6 cm in diameter, conventional bone grafts usually prove successful. However, for defects greater than 6 cm or in a poorly vascularized bed, the vascularized fibular graft can prevent such complications as fatigue fracture, failure of incorporation, and nonunion that are frequently seen with nonvascularized cortical and cancellous bone grafts.


Concomitant intestinal obstruction: a misleading diagnostic pitfall.
A 78-year-old man presented to the casualty department, complaining of recurrent and worsening constipation for the previous 2 months. This was associated with central, colicky abdominal pain and melena. In the last days, the symptoms worsened and the patient became partially obstructed, with nausea, vomiting and passing flatus but not stools for 72 h. The past medical history was unremarkable. The radiological findings of the plain abdominal film were consistent with mechanical small-bowel obstruction. CT scan revealed an intraluminal mass in the small bowel, which drew attention away from gross thickening of the caecal wall that was also present. A careful review of the images should not be omitted. One must be aware of a polymorphous appearance and the multiple causes of intestinal obstruction and avoid underestimating even the minor and less evident radiological findings.


Quantifying genomic connectedness and prediction accuracy from additive and non-additive gene actions.
Genetic connectedness is classically used as an indication of the risk associated with breeding value comparisons across management units because genetic evaluations based on best linear unbiased prediction rely for their success on sufficient linkage among different units. In the whole-genome prediction era, the concept of genetic connectedness can be extended to measure a connectedness level between reference and validation sets. However, little is known regarding (1) the impact of non-additive gene action on genomic connectedness measures and (2) the relationship between the estimated level of connectedness and prediction accuracy in the presence of non-additive genetic variation. We evaluated the extent to which non-additive kernel relationship matrices increase measures of connectedness and investigated its relationship with prediction accuracy in the cross-validation framework using best linear unbiased prediction and coefficients of determination. Simulated data assuming additive, dominance, and epistatic gene action scenarios and real swine data were analyzed. We found that the joint use of additive and non-additive genomic kernel relationship matrices or non-parametric relationship matrices led to increased capturing of connectedness, up to 25%, and improved prediction accuracies compared to those of baseline additive relationship counterparts in the presence of non-additive gene action. Our findings showed that connectedness metrics can be extended to incorporate non-additive genetic variation of complex traits. Use of kernel relationship matrices designed to capture non-additive gene action increased measures of connectedness and improved whole-genome prediction accuracy, further broadening the scope of genomic connectedness studies.


The use of theory in qualitative approaches to research: application in end-of-life studies.
This paper is a report of an analysis of the use of theory in qualitative approaches to research as exemplified in qualitative end-of-life studies. Nurses researchers turn to theory to conceptualize research problems and guide investigations. However, researchers using qualitative approaches do not consistently articulate how theory has been applied, and no clear consensus exists regarding the appropriate application of theory in qualitative studies. A review of qualitative, end-of-life studies is used to illustrate application of theory to study design and findings. A review of theoretical literature was carried out, focusing on definitions and use of theory in qualitative end-of-life studies published in English between 1990 and 2008. The term 'theory' continues to be used in a variety of ways by theorists and researchers. Within the reviewed end-of-life studies, the use of theory included theory creation or provision of a comparative framework for data analysis and interpretation. Implications for nursing. Nurses who conduct qualitative studies should examine the philosophical and theoretical bases of their selected methodological approach, articulate a theoretical framework that fits the phenomenon being studied, and adopt a critical, flexible and creative attitude when applying theory to a study. Theory can be put to several uses in qualitative inquiry and should guide nurse researchers as they develop and implement their studies. Nurse educators who teach qualitative approaches to research should emphasize a variety of ways to incorporate theory in qualitative designs.


Serum Hepcidin in Children with Inflammatory Bowel Disease.
Hepcidin is a major regulator of iron homeostasis and a mediator of innate immunity. To date, the role of hepcidin in inflammatory bowel disease (IBD) children is not clearly established. We aimed to assess serum hepcidin concentration in IBD children and correlate hepcidin with iron status parameters and inflammatory markers. The study group included 46 pediatric patients with ulcerative colitis and 29 with Crohn's disease. In control group, there were 21 children with functional gastrointestinal disorders. The complete blood count, high-sensitivity C-reactive protein, erythrocyte sedimentation rate, iron, ferritin, transferrin, hepcidin, soluble transferrin receptor, transferrin saturation, and interleukin-6 were measured. The study was approved by the local bioethical committee (KE-0254/22/2013). Mean serum hepcidin concentration was significantly decreased in IBD children (5.98 ng/mL) compared with controls (10 ng/mL) (P = 0.03). Hepcidin did not differ significantly between patients with Crohn's disease (6.9 ± 4.5 ng/mL) and ulcerative colitis (5.4 ± 5.3 ng/mL) (P = 0.07). Hepcidin was significantly decreased in IBD children with iron deficiency (4.9 ± 3.2 ng/mL) compared with healthy controls (10.5 ± 10 ng/mL) (P = 0.02). In anemic children with IBD, serum hepcidin (5.3 ± 4.4 ng/mL) was significantly reduced compared with healthy controls (10.5 ± 10 ng/mL) (P = 0.04), but comparable to nonanemic IBD children (6.6 ± 5.6 ng/mL) (P = 0.62). In IBD, children hepcidin was correlated solely with ferritin (P = 0.007; R = 0.3). In our study, serum hepcidin concentration was significantly decreased in IBD children compared with controls. Hepcidin correlated positively with ferritin, but not with any of inflammatory markers. It may suggest that in our cohort, hepcidin was regulated predominantly by iron storage level.


Complex Tumor Spheroids, a Tissue-Mimicking Tumor Model, for Drug Discovery and Precision Medicine.
Malignant tumors are complex tissues composed of malignant cells, vascular cells, structural mesenchymal cells including pericytes and carcinoma-associated fibroblasts, infiltrating immune cells, and others, collectively called the tumor stroma. The number of stromal cells in a tumor is often much greater than the number of malignant cells. The physical associations among all these cell types are critical to tumor growth, survival, and response to therapy. Most cell-based screens for cancer drug discovery and precision medicine validation use malignant cells in isolation as monolayers, embedded in a matrix, or as spheroids in suspension. Medium- and high-throughput screening with multiple cell lines requires a scalable, reproducible, robust cell-based assay. Complex spheroids include malignant cells and two normal cell types, human umbilical vein endothelial cells and highly plastic mesenchymal stem cells, which rapidly adapt to the malignant cell microenvironment. The patient-derived pancreatic adenocarcinoma cell line, K24384-001-R, was used to explore complex spheroid structure and response to anticancer agents in a 96-well format. We describe the development of the complex spheroid assay as well as the growth and structure of complex spheroids over time. Subsequently, we demonstrate successful assay miniaturization to a 384-well format and robust performance in a high-throughput screen. Implementation of the complex spheroid assay was further demonstrated with 10 well-established pancreatic cell lines. By incorporating both human stromal and tumor components, complex spheroids might provide an improved model for tumor response in vivo.


Role of human cytochrome P-450 IIE1 in the oxidation of many low molecular weight cancer suspects.
The role of human cytochrome P-450 IIE1 (P-450 IIE1) in the oxidation of a number of suspect carcinogens was examined by using a variety of approaches, including (1) selective inhibition of catalytic activity in human liver microsomes by diethyldithiocarbamate, which was found to be a selective mechanism-based inactivator of P-450 IIE1, (2) correlation of rates of different catalytic activities with each other and with chlorzoxazone 6-hydroxylation, an indicator of P-450 IIE1, in human liver microsomes, (3) demonstration of catalytic activity in reconstituted systems containing purified human P-450 IIE1, and (4) immunoinhibition of catalytic activity in human liver microsomes with rabbit anti-human P-450 IIE1. The results collectively indicate that P-450 IIE1 is a major catalyst of the oxidation of benzene, styrene, CCl4, CHCl3, CH2Cl2, CH3Cl, CH3CCl3, 1,2-dichloropropane, ethylene dichloride, ethylene dibromide, vinyl chloride, vinyl bromide, acrylonitrile, vinyl carbamate, ethyl carbamate, and trichloroethylene. Levels of P-450 IIE1 can vary considerably among individual humans--the availability of chlorzoxazone as a noninvasive probe of human P-450 IIE1 and of disulfiram (oxidized diethyldithiocarbamate) as an inhibitor may facilitate discernment of the in vivo significance of human P-450 IIE1 as a factor in the bioactivation and detoxication of these cancer suspects. Further, many investigations with diethyldithiocarbamate, disulfiram, and ethanol in humans and experimental animals may be interpreted in light of mechanisms involving P-450 IIE1.


Metabolism of 2-chloroethyl nitrosocarbamoylcystamine by rat liver subcellular fractions.
Previously we have shown that a new nitrosourea, 2-chloroethyl nitrosocarbamoylcystamine (CNCC), undergoes an extensive metabolism in the rat. Two pairs of plasma metabolites have been identified. This suggested our hypothesis that the metabolic pathway involves the reduction of the disulfur bridge followed by the methylation and the oxidation of the thiol groups. The two first intermediates, i.e. the unoxidized metabolites, could not be detected in vivo. Hence, to better understand and to confirm the proposed mechanism of biotransformation of CNCC, its in vitro metabolism has been studied. Incubation of CNCC with a rat liver homogenate or a 10,000g supernatant fraction leads to the formation of four pairs of metabolites. Among them we have identified the two first intermediates not found in vivo and the oxidized metabolites. These findings, together with the kinetics data, suggest that reduction, methylation, and oxidation are very rapid enzymatic reactions. We also show that, for completion of the reaction, the incubation mixture had to contain a cytosolic thioreductase, a microsomal and cytosolic S-methyltransferase, a microsomal oxidase, and an NADPH generating system. The sum of the amounts of metabolites found in the organic extratable material is less than the amount of CNCC metabolized. We conclude that the biotransformation of CNCC proceeds from two fast competitive mechanisms operative on both the disulfur and the nitroso groups.


Postoperative outcome after coronary artery bypass grafting in chronic obstructive pulmonary disease.
It is uncertain if the presence and severity of airflow obstruction in chronic obstructive pulmonary disease (COPD) is predictive of surgical morbidity and mortality after coronary artery bypass grafting (CABG). Retrospective study of patients who underwent CABG between 1998 and 2003 in a university-affiliated hospital for whom a preoperative spirometry was available. COPD was diagnosed in smokers or ex-smokers 50 years of age or older in the presence of irreversible airflow obstruction. Patients were divided into three groups depending on the spirometry: controls (forced expiratory volume in 1 s [FEV1] 80% or more, FEV1/forced vital capacity [FVC] greater than 0.7), mild to moderate COPD (FEV1 50% or more and FEV1/FVC 0.7 or less) and severe COPD (FEV1 less than 50% and FEV1/FVC 0.7 or less). Among the 411 files studied, 322 (249 men, 68+/-8 years of age) were retained (controls, n=101; mild to moderate COPD, n=153; severe COPD, n=68). The mortality rate (3.0%, 2.6% and 0%, respectively) was comparable among the three groups. Patients with severe COPD had a slightly longer hospital stay than controls (mean difference 0.7+/-1.4 days, P<0.05). Pulmonary infections were more frequent in severe COPD (26.5%) compared with mild to moderate COPD (12.4%) and controls (12.9%), P<0.05. Atrial fibrillation tended to be more frequent in severe COPD than in the other two groups. Mortality rate associated with CABG surgery is not influenced by the presence and severity of airflow obstruction in patients with COPD. The incidence of pulmonary infections and length of hospital stay were increased in patients with severe COPD.


Growth, blood, carcass and meat quality traits from local pig breeds and their crosses.
Contrary to intensive pig production, local pig breeds and their production systems are able to respond to the high criteria and expectations of modern society in regard to some environmental aspects, animal welfare, food quality and healthiness. This study proposes the recovery, study and use of a cross between two local breeds, contributing to animal biodiversity conservation and to the income of local pig producers. This work studied the growth performance and blood, carcass and meat quality traits of Alentejano (AL), Bísaro (BI) and Ribatejano (RI) (AL × BI, BI × AL) castrated male pigs. Raised outdoors, pigs were fed commercial diets ad libitum and killed at ~65 kg (trial 1, n = 10 from each genotype) and ~150 kg BW (trial 2, n = 9 from each genotype). In trial 1, AL and AL × BI attained slaughter weight later than BI and BI × AL pigs, with AL presenting lower average daily gains than the other genotypes (P < 0.001). Alentejano and RI pigs presented higher (P < 0.01) levels of plasma total protein than BI. Overall, carcass traits were affected by genotype, with length (P < 0.01), yield (P = 0.07) and lean cut proportions (P < 0.01) lower in AL than BI, and intermediate values for crossed pigs. Conversely, AL pigs presented higher fat cut proportion (P < 0.01), average backfat thickness (P < 0.001) and 'zwei punkte' fat depth (P < 0.01) than BI and RI pigs. Alentejano pigs also presented higher Longissimus lumborum (LL) intramuscular fat (P < 0.05), myoglobin content and ultimate pH (P < 0.01), but lower total collagen (P < 0.05), drip (P < 0.001) and cooking losses (P < 0.01), and shear force (P < 0.001) than all other genotypes. Finally, LL showed a more intense red colour in AL than in BI pigs. In trial 2, AL pigs confirmed to be a slow-growing obese breed with lower bone and lean cut proportions than BI, and higher LL intramuscular fat, richer colour, lower water loss and higher tenderness. In both trials, RI pigs grew faster, with higher lean and lower fat cut proportions and backfat thickness, and with overall LL characteristics comparable to those observed in AL pigs. This work demonstrates some clear differences between AL and BI breeds while showing that their crosses present intermediate characteristics in most studied traits. These data on RI pigs can be useful to breeders' associations and farmers in order to consider the use of these crosses as an option or complement to pure line breeding.


Characterization of a human placental fructose-6-phosphate, 2-kinase/fructose-2,6-bisphosphatase.
A full-length cDNA, which encodes a human placental fructose-6-phosphate,2-kinase/ fructose-2,6-bisphosphatase, was constructed and expressed in Escherichia coli. The expressed protein, purified to homogeneity, showed a molecular weight of 58,000 by gel electrophoresis under denaturing conditions, compared to the deduced molecular weight of 59,410. The N-terminal sequence of 15 amino acids coincided with that of the deduced sequence. The active enzyme was a dimer as judged by molecular sieve filtration. The expressed enzyme was bifunctional with Vmax values of 142 and 0.2 milliunits/mg for the kinase and phosphatase activities, respectively. The phosphatase activity was extremely low, because one phosphatase active site residue was mutated, and consequently the kinase/phosphatase ratio was the highest among the known isozymes. Furthermore, the enzyme was phosphorylated by cAMP-dependent protein kinase, protein kinase C and also by [2-32P]fructose-2,6-bisphosphate. Phosphorylation by cAMP-dependent protein kinase and protein kinase C increased the maximal Fru-6-P,2-kinase activities by 1.8- and 1.1-fold, respectively. These results suggested that placental fructose-6-phosphate,2-kinase/ fructose-2,6-bisphosphatase is important in maintaining and regulating a relatively high rate of glycolysis in placenta.


Tunable focusing of sine-azimuthal wavefront modulated cosh-Gaussian beams by one spiral optical vortex.
Optical intensity distributions in the focal region play an important role in many optical systems. In this paper, the tunable focusing properties of linearly polarized hyperbolic-cosine-Gaussian beams with sine-azimuthal variation wavefront were investigated by adding a spiral optical vortex. It was found that the focal patterns can be altered very considerably by changing the charge number of the optical vortex under a different phase parameter that indicates the phase change frequency upon increasing the azimuthal angle. The symmetry of focal patterns also changes remarkably upon increasing the charge number. And some novel focal patterns may appear, including a multiple-peak array, wheel focal pattern, or swallow-tailed focal pattern.


Sodium currents in naïve mouse dorsal root ganglion neurons: No major differences between sexes.
Sexual dimorphism has been reported in multiple pre-clinical and clinical studies on pain. Previous investigations have suggested that in at least some states, rodent dorsal root ganglion (DRG) neurons display differential sex-dependent regulation and expression patterns of various proteins involved in the pain pathway. Our goal in this study was to determine whether sexual dimorphism in the biophysical properties of voltage-gated sodium (Nav) currents contributes to these observations in rodents. We recently developed a novel method that enables high-throughput, unbiased, and automated functional analysis of native rodent sensory neurons from naïve WT mice profiled simultaneously under uniform experimental conditions. In our previous study, we performed all experiments in neurons that were obtained from mixed populations of adult males or females, which were combined into single (combined male/female) data sets. Here, we have re-analyzed the same previously published data and segregated the cells based on sex. Although the number of cells in our previously published data sets were uneven for some comparisons, our results do not show sex-dependent differences in the biophysical properties of Nav currents in these native DRG neurons.


Temporal dynamics and decay of putatively allochthonous and autochthonous viral genotypes in contrasting freshwater lakes.
Aquatic viruses play important roles in the biogeochemistry and ecology of lacustrine ecosystems; however, their composition, dynamics, and interactions with viruses of terrestrial origin are less extensively studied. We used a viral shotgun metagenomic approach to elucidate candidate autochthonous (i.e., produced within the lake) and allochthonous (i.e., washed in from other habitats) viral genotypes for a comparative study of their dynamics in lake waters. Based on shotgun metagenomes prepared from catchment soil and freshwater samples from two contrasting lakes (Cayuga Lake and Fayetteville Green Lake), we selected two putatively autochthonous viral genotypes (phycodnaviruses likely infecting algae and cyanomyoviruses likely infecting picocyanobacteria) and two putatively allochthonous viral genotypes (geminiviruses likely infecting terrestrial plants and circoviruses infecting unknown hosts but common in soil libraries) for analysis by genotype-specific quantitative PCR (TaqMan) applied to DNAs from viruses in the viral size fraction of lake plankton, i.e., 0.2 μm > virus > 0.02 μm. The abundance of autochthonous genotypes largely reflected expected host abundance, while the abundance of allochthonous genotypes corresponded with rainfall and storm events in the respective catchments, suggesting that viruses with these genotypes may have been transported to the lake in runoff. The decay rates of allochthonous and autochthonous genotypes, assessed in incubations where all potential hosts were killed, were generally lower (0.13 to 1.50% h(-1)) than those reported for marine virioplankton but similar to those for freshwater virioplankton. Both allochthonous and autochthonous viral genotypes were detected at higher concentrations in subsurface sediments than at the water-sediment interface. Our data indicate that putatively allochthonous viruses are present in lake plankton and sediments, where their temporal dynamics reflect active transport to the lake during hydrological events and then decay once there.


Peak deformations in preparative supercritical fluid chromatography due to co-solvent adsorption.
In supercritical fluid chromatography (SFC) the mobile phase comprises of carbon dioxide (CO2) as main solvent and smaller amounts of an organic polar solvent (often an alcohol) as co-solvent. The co-solvent is considered to function by changing the overall polarity of the eluent, i.e. by acting as a "modifier". However, recent studies indicate that the co-solvent methanol can also adsorb to some common SFC stationary phases. Hence, the co-solvent should also be able to function as an "adsorbing additive", i.e. an eluent component that competes with the injected solutes about the stationary phase surface. In this study it was found by fitting different mechanistic models to systematic experimental data, that the co-solvent methanol can have both functions: at low co-solvent fractions, methanol acts as an additive whereas at larger fractions it acts as a modifier. Moreover, it was found that when the co-solvent adsorbs more strongly to the stationary phase than the solute, "bizarre" deformations of the preparative band shapes can occur. This is illustrated by a solute that converts from a normal "Langmuirian" band shape to an "anti-Langmuirian" shape when changing from neat carbon dioxide (CO2) to an eluent containing co-solvent. This peak shape transition is dependent on both (i) the relative retention of the solute and co-solvent to the stationary phase in eluent containing neat CO2 and on (ii) the relative retention of the additive perturbation peak and the solute peak in eluent containing also co-solvent.


Development of the social brain in adolescence.
The brain has evolved to understand and interact with other people. We are increasingly learning more about the neurophysiological basis of social cognition and what is known as the social brain, that is the network of brain regions involved in understanding others. This paper focuses on how the social brain develops during adolescence. Adolescence is a time characterized by change - hormonally, physically, psychologically and socially. Yet until recently this period of life was neglected by cognitive neuroscience. In the past decade, research has shown that the brain develops both structurally and functionally during adolescence. Large-scale structural MRI studies have demonstrated development during adolescence in white matter and grey matter volumes in regions within the social brain. Activity in some of these regions, as measured using fMRI, also shows changes between adolescence and adulthood during social cognition tasks. I will also present evidence that theory of mind usage is still developing late in adolescence. Finally, I will speculate on potential implications of this research for society.


Catching a floating thrombus; a case report on the treatment of a large thrombus in the ascending aorta.
The ascending aorta is an uncommon site for non-infective thrombus. In non-aneurysmal and non-atherosclerotic vessels this condition becomes extremely rare, while it represents a source of potential cerebral and peripheral embolic events. Currently, there is no consensus in the guidelines on how to treat a free floating thrombus in ascending aorta, therefore we present our decision making process and therapeutic strategy. A healthy 48-year-old man was hospital admitted with acute abdominal pain. CT-scan showed a right renal embolism in presence of a defect in the distal ascending aorta suggestive for thrombus. After heart team discussion the patient was scheduled for surgery and successfully underwent an emergent thrombus removal. Also, owing to multiple aortic wall insertions, the ascending aorta was replaced. The patient's recovery was uneventful and histological examination showed no signs of connective tissue disorders of aortic wall while confirmed the thrombotic nature of the mass. We present a patient with a floating thrombus in the ascending aorta who underwent an ascending aorta replacement. While angio-CT scan led to a prompt diagnosis, intraoperative epi-aortic echocardiography allowed to define precise location of thrombus, minimizing operative risk. This case demonstrates that multi-disciplinary heart team discussion is essential to define a successful strategy, that surgical treatment is feasible with specific tools such as epi-aortic echocardiography.


Postresuscitation disease.
The postresuscitation disease is a specific pathophysiologic state of vital organ systems early after ischemic anoxia. This report summarizes reviews of past research and makes suggestions for future research concerning revival of the cerebral cortex after clinical death, CNS stimulation vs. sedation, postischemic coma and pain, near-death experiences, and extracerebral derangements. The stages of resuscitation when the CNS should be stimulated and those when it is preferable to depress the activity of not fully recovered higher centers remain to be clarified. Future research in reanimatology should include the chemical nature of endotoxins in terminal states. Adult respiratory distress syndrome (ARDS, shock lung), a component of the postresuscitation disease, occurs frequently after cardiac arrest or in sepsis and cannot be fully prevented by artificial ventilation. Prevention of ARDS should also be studied.


[Guidelines of the Italian Society of Nephrology. Peritoneal dialysis Guidelines].
The adequate dosage of peritoneal dialysis (PD) has been defined (using unrandomized and uncontrolled studies) asKt/V = 2, with a total (peritoneal + renal) creatinine clearance = 60 mL/min. The recent prospective, randomized ADEMEX study, suggests targets of 1.8 and 54 mL/min respectively. Dialysis must also be adequate to control fluid removal, phosphate levels, nutritional status, and hypertension. The targets for automated PD (APD) should be either 10% more than CAPD or similar, depending on the time of blood sample collection either immediately at the end of the automated exchanges or 6 to 8 hours after. A peritoneal equilibration test should be done 1 to 2 months after the start of PD, yearly, and when peritoneal permeability or ultrafiltration changes occurr. Residual renal function must be protected as long as possible by avoiding nephrotoxic drugs and excessive dehydration. Every effort must be taken in the attempt to maintain a good nutritional status and to diagnose as soon as possible any changes toward malnutrition. Hypertension has a high prevalence in PD patients and has negative effects on both cardiovascular status and patient survival. However, anti-hypertensive therapy should avoid hypotension, mainly in older patients, who are more at risk for cerebrovascular accident. Hyperparathiroidism must be controlled by diet, phosphate binders, and calcitriol supplement, but attention must be paid to avoid cardiac and vascular calcifications. Peritonitis and exit-site infection should be prevented by all means available. In the case of infection, empiric antibiotic therapy should be started as soon as possible and then adapted according to the antibiogram.


Facile Synthesis and Characterization of a Novel Tamavidin-Luciferase Reporter Fusion Protein for Universal Signaling Applications.
Despite the avidin/biotin reaction being one of the most ubiquitous noncovalent immobilization and sensing strategies in scientific research, the ability to synthesize useful amounts of biotin-binding fusion constructs is hampered by poor solubility in bacterial expression systems. As such, there are few reports of successful genetic reporter fusions incorporating a biotin-binding partner. To address this, a sensitivity-enhanced, synthetically facile reporter fusion is developed to merge the bioluminescence output of Gaussia luciferase (Gluc) with the recently characterized biotin-binding ability of tamavidin 2 (TA2) for general and universal signaling applications in biological and analytical systems. This fusion construct enables direct bacterial expression of a reporter system incorporating two important functionalities in a 1:1 stoichiometric relationship that can provide detection of discrete events at low concentrations. Using a cold-shock expression system, highly concentrated construct can be obtained from standard culture volumes while retaining essentially native protein activity. To demonstrate feasibility and provide an example application, this fusion construct is then included in a standard target-bridged assay design for the sensitive detection of four miRNA targets.


[A method of determining the efficiency of the projected process of uninterrupted microorganism cultivation for vaccine production].
The authors suggest the method of study of the process of periodic cultivation of microorganisms for prognostication of the optimal process of biomass accumulation at the level of microbial population, homogenous by the spectrum of biochemical properties. A regimen of continuous growth of the microorganisms (calculated with the use of this method), corresponding to the phase of delay of the growth rate, was economically more effective in comparison with the traditional process used in the vaccine production.


Exploring the complex response to linuron of bacterial communities from biopurification systems by means of cultivation-independent methods.
On-farm biopurification systems (BPSs) treat pesticide-contaminated wastewater at farms through biodegradation and sorption processes. However, information on the microbiota involved in pesticide removal in BPSs is scarce. Here we report on the response of BPS bacterial communities to the herbicide linuron (BPS(+)) compared with the control (BPS(-)) in a microcosm experiment. Both denaturing gradient gel electrophoresis (DGGE) and pyrosequencing of 16S rRNA gene fragments amplified from community DNA indicated shifts in the bacterial community after linuron application. Responding populations belonged to taxa that were previously reported from linuron degrading consortia cultivated from soil (Hyphomicrobiaceae, Comamonadaceae, Micrococcaceae). In addition, numerous taxa with increased relative abundance were identified that were previously not associated with linuron degradation. The relative abundance of IncP-1 korB copies increased in response to linuron application. Amplicon pyrosequencing of IncP-1 trfA genes revealed a high IncP-1 plasmid diversity and suggested that populations carrying IncP-1β plasmids increased in relative abundance. Transferable mercury resistance plasmids were exogenously captured from BPS(+)/BPS(-), and in three transconjugants from BPS(+) the gene hylA was detected. Our data suggest the existence of a multispecies linuron degrading bacterial food web and an involvement of IncP-1 plasmids in the adaptation of bacterial communities to pesticide pollution in BPSs.


The Roles of Coenzyme A Binding Pocket Residues in Short and Medium Chain Acyl-CoA Synthetases.
Short- and medium-chain acyl-CoA synthetases catalyze similar two-step reactions in which acyl substrate and ATP bind to form an enzyme-bound acyl-adenylate, then CoA binds for formation of the acyl-CoA product. We investigated the roles of active site residues in CoA binding in acetyl-CoA synthetase (Acs) and a medium-chain acyl-CoA synthetase (Macs) that uses 2-methylbutyryl-CoA. Three highly conserved residues, Arg193, Arg528, and Arg586 of Methanothermobacter thermautotrophicus Acs (AcsMt), are predicted to form important interactions with the 5'- and 3'-phosphate groups of CoA. Kinetic characterization of AcsMt variants altered at each of these positions indicates these Arg residues play a critical role in CoA binding and catalysis. The predicted CoA binding site of Methanosarcina acetivorans Macs (MacsMa) is structurally more closely related to that of 4-chlorobenzoate:coenzyme A ligase (CBAL) than Acs. Alteration of MacsMa residues Tyr460, Arg490, Tyr525, and Tyr527, which correspond to CoA binding pocket residues in CBAL, strongly affected CoA binding and catalysis without substantially affecting acyl-adenylate formation. Both enzymes discriminate between 3'-dephospho-CoA and CoA, indicating interaction between the enzyme and the 3'-phosphate group is important. Alteration of MacsMa residues Lys461 and Lys519, located at positions equivalent to AcsMt Arg528 and Arg586, respectively, had only a moderate effect on CoA binding and catalysis. Overall, our results indicate the active site architecture in AcsMt and MacsMa differs even though these enzymes catalyze mechanistically similar reactions. The significance of this study is that we have delineated the active site architecture with respect to CoA binding and catalysis in this important enzyme superfamily.


A potential role for plasma uric acid in the endothelial pathology of Plasmodium falciparum malaria.
Inflammatory cytokinemia and systemic activation of the microvascular endothelium are central to the pathogenesis of Plasmodium falciparum malaria. Recently, 'parasite-derived' uric acid (UA) was shown to activate human immune cells in vitro, and plasma UA levels were associated with inflammatory cytokine levels and disease severity in Malian children with malaria. Since UA is associated with endothelial inflammation in non-malaria diseases, we hypothesized that elevated UA levels contribute to the endothelial pathology of P. falciparum malaria. We measured levels of UA and soluble forms of intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1), E-selectin (sE-Selectin), thrombomodulin (sTM), tissue factor (sTF) and vascular endothelial growth factor (VEGF) in the plasma of Malian children aged 0.5-17 years with uncomplicated malaria (UM, n = 487) and non-cerebral severe malaria (NCSM, n = 68). In 69 of these children, we measured these same factors once when they experienced a malaria episode and twice when they were healthy (i.e., before and after the malaria transmission season). We found that levels of UA, sICAM-1, sVCAM-1, sE-Selectin and sTM increase during a malaria episode and return to basal levels at the end of the transmission season (p<0.0001). Plasma levels of UA and these four endothelial biomarkers correlate with parasite density and disease severity. In children with UM, UA levels correlate with parasite density (r = 0.092, p = 0.043), sICAM-1 (r = 0.255, p<0.0001) and sTM (r = 0.175, p = 0.0001) levels. After adjusting for parasite density, UA levels predict sTM levels. Elevated UA levels may contribute to malaria pathogenesis by damaging endothelium and promoting a procoagulant state. The correlation between UA levels and parasite densities suggests that parasitized erythrocytes are one possible source of excess UA. UA-induced shedding of endothelial TM may represent a novel mechanism of malaria pathogenesis, in which activated thrombin induces fibrin deposition and platelet aggregation in microvessels. This protocol is registered at clinicaltrials.gov (NCT00669084).


Quality of life in Nepalese Patients with Melasma: An Observational Cross-Sectional Study at a Tertiary Center.
Melasma is an acquired pigmentation disorder of the sun exposed parts of the body, particularly face. It is a significant cosmetic concern for the young adults, which is the most frequently affected age-group. Finding the level of impairment in quality of life in melasma patients and correlating with the severity. All consenting patients with melasma visiting our out-patient from December 2020 to June 2021 were enrolled. Clinical evaluation was done by the same single reviewer in all cases using modified Melasma Area and Severity Index. Dermatology Life Quality Index (Nepali version) was asked to complete to the patient. Data were collected and analyzed. Frequency and mean were calculated for all variables, comparison of means by Mann-Whitney U test and correlation analyses by Spearman's correlation test were performed. Females were 82.5%(99) of total cases. Among these 70.8%(85) were married, 68.3%(82) had family history of melasma and 65%(78) had centro-facial phenotype. Mean Dermatology Life Quality Index score was 10.25±0.54. Mean score of females was higher than that of males. The daily activities sub-domain was the worst affected with 50.67% of impairment. Females suffered significantly more than males in symptom and feeling sub-domain. Modified Melasma Area and Severity Index was significantly higher in males (7.12±.56 Vs. 5.66±.28). The severity and duration of melasma did not correlate to the quality of life. Melasma had a moderately negative impact in the quality of life of the patients. Females suffered more due to melasma. The clinical severity of melasma did not correlate with the quality of life impairment. Thus, psychological assessment to all the melasma patients seem to be an important aspect of management.


Prevalence of 25 canine behavioral problems and relevant factors of each behavior in Japan.
Undesirable behaviors exhibited by dogs are called canine behavioral problems. Behavioral problems are common among dogs kept in ordinary homes; hence, veterinarians need to have sufficient knowledge of these problems, so as to advise owners. Among dogs kept in ordinary Japanese homes, only a few behaviors have been investigated regarding prevalence and relevant factors. To improve understanding of behavioral problems, a comprehensive survey is crucial. In this study, we distributed an online survey including general information and questions about 25 canine behaviors that can become problematic to owners, to investigate (1) the prevalence of behavioral problem, (2) the most common behavioral problems, and (3) the relevant factors of each behavior. We collected 2,050 valid responses. Among all dogs, 86.0% exhibited at least one behavioral problem. "Barking at noises inside the house", "Barking at unfamiliar visitors", and "Pica" were the most frequent behavioral problems. To reveal those factors related to the expression of each behavior, we performed multiple logistic regression analyses, with the presence of behaviors as objective variables and dogs' general information as explanatory variables. Breed, age, the presence of other dogs in the household, sex, neutering status, source of acquisition, and type of residence were associated with 19, 16, 5, 4, 4, 4, and 1 behaviors, respectively. This study revealed the prevalence of 25 canine behavioral problems in Japan and relevant factors of each behavior. Our findings will be useful for veterinarians in advising owners about preventing behavioral problems.


Loss of SUMO-specific protease 2 causes isolated glucocorticoid deficiency by blocking adrenal cortex zonal transdifferentiation in mice.
SUMOylation is a dynamic posttranslational modification, that provides fine-tuning of protein function involved in the cellular response to stress, differentiation, and tissue development. In the adrenal cortex, an emblematic endocrine organ that mediates adaptation to physiological demands, the SUMOylation gradient is inversely correlated with the gradient of cellular differentiation raising important questions about its role in functional zonation and the response to stress. Considering that SUMO-specific protease 2 (SENP2), a deSUMOylating enzyme, is upregulated by Adrenocorticotropic Hormone (ACTH)/cAMP-dependent Protein Kinase (PKA) signalling within the zona fasciculata, we generated mice with adrenal-specific Senp2 loss to address these questions. Disruption of SENP2 activity in steroidogenic cells leads to specific hypoplasia of the zona fasciculata, a blunted reponse to ACTH and isolated glucocorticoid deficiency. Mechanistically, overSUMOylation resulting from SENP2 loss shifts the balance between ACTH/PKA and WNT/β-catenin signalling leading to repression of PKA activity and ectopic activation of β-catenin. At the cellular level, this blocks transdifferentiation of β-catenin-positive zona glomerulosa cells into fasciculata cells and sensitises them to premature apoptosis. Our findings indicate that the SUMO pathway is critical for adrenal homeostasis and stress responsiveness.


Postal self-exposure treatment of recurrent nightmares. Randomised controlled trial.
Many nightmare sufferers do not consult a health care professional. Though behavior and cognitive therapy can help, they have not been tested as a self-treatment method at home using a manual. One hundred and seventy adults with primary nightmares were randomised to four weeks' self-exposure or self-relaxation at home using manuals posted to them, or to a waiting-list as a control group for four weeks. Individuals recorded nightmare frequency and intensity in four-week diaries. At one- and six-month follow-up, the self-rated nightmare frequency fell more significantly in exposure subjects than relaxation or waiting-list subjects. The self-exposure group had the most drop-outs but remained superior in an over-cautious intent-to-treat analysis. The individuals' partners confirmed the superiority of self-exposure to self-relaxation at one- and six-month follow-up. Recurrent nightmare sufferers improved more with self-exposure manuals than with self-relaxation manuals or by being on a waiting-list. Self-exposure may be needed for longer than four weeks in order to reduce nightmare intensity as well as frequency. Despite a high drop-out rate, some sufferers of other conditions may benefit from self-treatment manuals.


Wellbeing, Whole Health and Societal Transformation: Theoretical Insights and Practical Applications.
Society faces several major interrelated challenges which have an increasingly profound impact on global health including inequalities, inequities, chronic disease and the climate catastrophe. We argue here that a focus on the determinants of wellbeing across multiple domains offers under-realised potential for promoting the 'whole health' of individuals, communities and nature. Here, we review recent theoretical innovations that have laid the foundations for our own theoretical model of wellbeing - the GENIAL framework - which explicitly links health to wellbeing, broadly defined. We emphasise key determinants across multiple levels of scale spanning the individual, community and environmental levels, providing opportunities for positive change that is either constrained or facilitated by a host of sociostructural factors lying beyond the immediate control of the individual (e.g. social cohesion and health-related inequities can either promote or adversely impact on wellbeing, respectively). Following this, we show how the GENIAL theoretical framework has been applied to various populations including university students and people living with neurological disorders, with a focus on acquired brain injury. The wider implication of our work is discussed in terms of its contribution to the understanding of 'whole health' as well as laying the foundations for a 'whole systems' approach to improving health and wellbeing in a just and sustainable way.


Cortical reorganization in malformations of cortical development: a magnetoencephalographic study.
The evaluation for epilepsy surgery of patients with malformations of cortical development (MCDs) in areas of clinically important cerebral function is a challenge because of the unpredictable localization of critical sensory, motor, and cognitive function. Magnetoencephalography (MEG) source localization of evoked fields can address whether functional reorganization of primary sensory modalities exists in MCDs. Consecutive patients with MRI-demonstrated rolandic and calcarine cortex MCDs were identified who had a 148-channel whole-head MEG study to identify the localization of primary somatosensory and visual cortices. Reorganization was considered when localization contrasted that expected upon general anatomic or homuncular rules and was defined against controls. Twelve patients (n = 12) were studied. Six had focal cortical dysplasia, two had polymicrogyria and schizencephaly, and four had isolated polymicrogyria. In the patients with cortical dysplasias, the somatosensory cortices were identified outside the rolandic area. In the two patients with polymicrogyria and schizencephaly, the somatosensory cortices remained in the rolandic areas as long as the anatomy was not distorted by the presence of the schizencephalic cleft. In the patients with isolated polymicrogyria, the somatosensory cortex was mapped without evidence of reorganization. In patients with epileptic MCDs involving rolandic and calcarine regions, cortical function may be reorganized if the MCDs are due to an abnormal neuronal or glial proliferation (i.e., cortical dysplasia) but may not be in MCDs caused by abnormal cortical organization (i.e., polymicrogyria).


Optical absorption in complexes of abasic DNA with noble-metal nanoclusters by first principles calculations.
Abasic sites (AP site) in a DNA duplex have been experimentally used to produce fluorescent Ag nanoclusters (NC) with a small number of atoms (n ≤ 6). These AP-DNA:NC complexes act as biological makers that help to locate genes associated with diseases related to single nucleotide polymorphisms (SNP), for example. Abasic sites are the most common SNP genetic variation, and their detection may help predict a host of genetically determined diseases. In this work, we report a theoretical study of the optical absorption spectra of AP-DNA:Ag4 and AP-DNA:Au4 complexes using a fully ab initio methodology. We consider several different base environments for the noble-metal nanocluster occupying the AP site, and compute the absorption spectra of sixteen AP-DNA:Ag4 and sixteen AP-DNA:Au4 complexes. We find that optical absorption in the AP-DNA:Ag4 complexes tends to concentrate in the green-to-violet range of frequencies (2.50 eV ≤ ħω ≤ 3.2 eV) and that AP-DNA:Au4 complexes display absorption peaks in the violet-to-ultraviolet interval (ħω ≥ 3.0 eV). An analysis of the optical absorption mechanisms in these complexes shows that they can be of local, charge-transfer, or hybrid nature, i.e., AP-DNA:NC complexes display the full variety of optical absorption processes in molecular systems. In particular, we identify both charge-transfer and hybrid processes involving several DNA bases surrounding the NC. Importantly, we find that even sequences where the Ag4 cluster is not in a guanine rich neighborhood display absorption peaks in the visible-light spectrum. Moreover, we obtain that the maximum intensities of the absorption peaks in complexes with pyrimidine vacancies are generally higher than those in complexes with purine vacancies. Regarding the selectivity of single-vacancy AP-DNA to specific noble-metal nanocluster sizes, our calculations show that the four-atom Ag4 (Au4) species fits naturally and binds into the AP-site in a single-vacancy AP-DNA.


Folic acid-functionalized chitosan nanoparticles with bioenzyme activity for the treatment of spinal cord injury.
Spinal cord injury (SCI) is a central system disease with a high rate of disability. Pathological changes such as ischemia and hypoxia of local tissues, oxidative stress and apoptosis could lead to limb pain, paralysis and even life-threatening. It was reported that catalase (CAT) was the main antioxidant in organisms, which could remove reactive oxygen species (ROS) and release oxygen (O2). However, the efficacy of the drug is largely limited due to its poor stability, low bioavailability and inability to cross the blood spinal cord barrier (BSCB). Therefore, in this study, we prepared folic acid-functionalized chitosan nanoparticles to deliver CAT (FA-CSNCAT) for solving this problem. In vivo small animal imaging results showed that FA-CSN could carry CAT across the BSCB and target to the inflammatory site. In addition, Immunofluorescence, ROS assay and JC-1 probe were used to detect the therapeutic effect of FA-CSNCAT in vitro and in vivo. The results showed that FA-CSNCAT could alleviate the hypoxic environment at the injured site and remove ROS, thereby inhibiting oxidative stress and protecting neurons, which may provide a new idea for clinical medication of SCI.


Medicare HMOs: part of the solution or part of the problem in the struggle for cost containment?
The correct answer for salvage of the Medicare program while maintaining acceptable medical care is clearly not available today. Managed care programs for seniors have been an option in the last decade and plans are expanding rapidly into previously untapped areas. Some programs can indeed provide a multiplicity of services and thereby truly integrate and improve existing care plans. Others have less clear cut advantages. For any senior contemplating the options available for medical care under Medicare, HMOs and Medigap type instruments, the most salient advice is "buyer beware." A complete understanding of the options gained and lost by changing medical insurance coverage is essential.


A sustained release formulation of novel quininib-hyaluronan microneedles inhibits angiogenesis and retinal vascular permeability in vivo.
Pathologic neovascularisation and ocular permeability are hallmarks of proliferative diabetic retinopathy and age-related macular degeneration. Current pharmacologic interventions targeting VEGF are effective in only 30-60% of patients and require multiple intraocular injections associated with iatrogenic infection. Thus, our goal is to develop novel small molecule drugs that are VEGF-independent are amenable to sustained ocular-release, and which reduce retinal angiogenesis and retinal vascular permeability. Here, the anti-angiogenic drug quininib was formulated into hyaluronan (HA) microneedles whose safety and efficacy was evaluated in vivo. Quininib-HA microneedles were formulated via desolvation from quininib-HA solution and subsequent cross-linking with 4-arm-PEG-amine prior to freeze-drying. Scanning electron microscopy revealed hollow needle-shaped particle ultrastructure, with a zeta potential of -35.5mV determined by electrophoretic light scattering. The incorporation efficiency and pharmacokinetic profile of quininib released in vitro from the microneedles was quantified by HPLC. Quininib incorporation into these microneedles was 90%. In vitro, 20% quininib was released over 4months; or in the presence of increasing concentrations of hyaluronidase, 60% incorporated quininib was released over 4months. Zebrafish hyaloid vasculature assays demonstrated quininib released from these microneedles significantly (p<0.0001) inhibited ocular developmental angiogenesis compared to control. Sustained amelioration of retinal vascular permeability (RVP) was demonstrated using a bespoke cysteinyl leukotriene induced rodent model. Quininib-HA microparticles significantly inhibited RVP in Brown Norway rats one month after administration compared to neat quininib control (p=0.0071). In summary, quininib-HA microneedles allow for sustained release of quininib; are safe in vivo and quininib released from these microneedles effectively inhibits angiogenesis and RVP in vivo.


Methanogenic Biocathode Microbial Community Development and the Role of Bacteria.
The cathode microbial community of a methanogenic bioelectrochemical system (BES) is key to the efficient conversion of carbon dioxide (CO2) to methane (CH4) with application to biogas upgrading. The objective of this study was to compare the performance and microbial community composition of a biocathode inoculated with a mixed methanogenic (MM) culture to a biocathode inoculated with an enriched hydrogenotrophic methanogenic (EHM) culture, developed from the MM culture following pre-enrichment with H2 and CO2 as the only externally supplied electron donor and carbon source, respectively. Using an adjacent Ag/AgCl reference electrode, biocathode potential was poised at -0.8 V (versus SHE) using a potentiostat, with the bioanode acting as the counter electrode. When normalized to cathode biofilm biomass, the methane production in the MM- and EHM-biocathode was 0.153 ± 0.010 and 0.586 ± 0.029 mmol CH4/mg biomass-day, respectively. This study showed that H2/CO2 pre-enriched inoculum enhanced biocathode CH4 production, although the archaeal communities in both biocathodes converged primarily (86-100%) on a phylotype closely related to Methanobrevibacter arboriphilus. The bacterial community of the MM-biocathode was similar to that of the MM inoculum but was enriched in Spirochaetes and other nonexoelectrogenic, fermentative Bacteria. In contrast, the EHM-biocathode bacterial community was enriched in Proteobacteria, exoelectrogens, and putative producers of electron shuttle mediators. Similar biomass levels were detected in the MM- and EHM-biocathodes. Thus, although the archaeal communities were similar in the two biocathodes, the difference in bacterial community composition was likely responsible for the 3.8-fold larger CH4 production rate observed in the EHM-biocathode. Roles for abundant OTUs identified in the biofilm and inoculum cultures were highlighted on the basis of previous reports.


Antiviral activity of the exopolysaccharide produced by Serratia sp. strain Gsm01 against Cucumber mosaic virus.
The potential of the exopolysaccharide (EPS) from a Serratia sp. strain Gsm01 as an antiviral agent against a yellow strain of Cucumber mosaic virus (CMV-Y) was evaluated in tobacco plants (Nicotiana tabacum cv. Xanthi-nc). The spray treatment of plants using an EPS preparation, 72 before CMV-Y inoculation, protected them against symptom appearance. Fifteen days after challenge inoculation with CMVY, 33.33% of plants showed mosaic symptoms in EPS-treated plants compared with 100% in the control plants. The EPS-treated plants, which showed mosaic symptoms, appeared three days later than the controls. The enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) analyses of the leaves of the protected plants revealed that the EPS treatment affected virus accumulation in those plants. Analysis of phenylalanine ammonia lyase, peroxidase, and phenols in protected plants revealed enhanced accumulation of these substances. The pathogenesis-related (PR) genes expression represented by PR-1b was increased in EPS-treated plants. This is the first report of a systemic induction of protection triggered by EPS produced by Serratia sp. against CMV-Y.


The demographic impact of extreme events: stochastic weather drives survival and population dynamics in a long-lived seabird.
1. Most scenarios for future climate change predict increased variability and thus increased frequency of extreme weather events. To predict impacts of climate change on wild populations, we need to understand whether this translates into increased variability in demographic parameters, which would lead to reduced population growth rates even without a change in mean parameter values. This requires robust estimates of temporal process variance, for example in survival, and identification of weather covariates linked to interannual variability. 2. The European shag Phalacrocorax aristotelis (L.) shows unusually large variability in population size, and large-scale mortality events have been linked to winter gales. We estimated first-year, second-year and adult survival based on 43 years of ringing and dead recovery data from the Isle of May, Scotland, using recent methods to quantify temporal process variance and identify aspects of winter weather linked to survival. 3. Survival was highly variable for all age groups, and for second-year and adult birds process variance declined strongly when the most extreme year was excluded. Survival in these age groups was low in winters with strong onshore winds and high rainfall. Variation in first-year survival was not related to winter weather, and process variance, although high, was less affected by extreme years. A stochastic population model showed that increasing process variance in survival would lead to reduced population growth rate and increasing probability of extinction. 4. As in other cormorants, shag plumage is only partially waterproof, presumably an adaptation to highly efficient underwater foraging. We speculate that this adaptation may make individuals vulnerable to rough winter weather, leading to boom-and-bust dynamics, where rapid population growth under favourable conditions allows recovery from periodic large-scale weather-related mortality. 5. Given that extreme weather events are predicted to become more frequent, species such as shags that are vulnerable to such events are likely to exhibit stronger reductions in population growth than would be expected from changes in mean climate. Vulnerability to extreme events thus needs to be accounted for when predicting the ecological impacts of climate change.


Acute Dietary Nitrate Intake Improves Muscle Contractile Function in Patients With Heart Failure: A Double-Blind, Placebo-Controlled, Randomized Trial.
Skeletal muscle strength, velocity, and power are markedly reduced in patients with heart failure, which contributes to their impaired exercise capacity and lower quality of life. This muscle dysfunction may be partially because of decreased nitric oxide (NO) bioavailability. We therefore sought to determine whether ingestion of inorganic nitrate (NO3 (-)) would increase NO production and improve muscle function in patients with heart failure because of systolic dysfunction. Using a double-blind, placebo-controlled, randomized crossover design, we determined the effects of dietary NO3 (-) in 9 patients with heart failure. After fasting overnight, subjects drank beetroot juice containing or devoid of 11.2 mmol of NO3 (-). Two hours later, muscle function was assessed using isokinetic dynamometry. Dietary NO3 (-) increased (P<0.05-0.001) breath NO by 35% to 50%. This was accompanied by 9% (P=0.07) and 11% (P<0.05) increases in peak knee extensor power at the 2 highest movement velocities tested (ie, 4.71 and 6.28 rad/s). Maximal power (calculated by fitting peak power data with a parabola) was therefore greater (ie, 4.74±0.41 versus 4.20±0.33 W/kg; P<0.05) after dietary NO3 (-) intake. Calculated maximal velocity of knee extension was also higher after NO3 (-) ingestion (ie, 12.48±0.95 versus 11.11±0.53 rad/s; P<0.05). Blood pressure was unchanged, and no adverse clinical events occurred. In this pilot study, acute dietary NO3 (-) intake was well tolerated and enhanced NO bioavailability and muscle power in patients with systolic heart failure. Larger-scale studies should be conducted to determine whether the latter translates into an improved quality of life in this population. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01682356.


Poor outcome of revised resurfacing hip arthroplasty.
Recent years have seen a rapid increase in the use of resurfacing hip arthroplasty despite the lack of literature on the long-term outcome. In particular, there is little evidence regarding the outcome of revisions of primary resurfacing. The purpose of this analysis was to examine the survivorship of primary resurfacing hip arthroplasties that have been revised. Over 12,000 primary resurfacing hip arthroplasties were recorded by the Australian Orthopaedic Association National Joint Replacement Registry between September 1, 1999 and December 31, 2008. During this time, 397 revisions for reasons other than infection were reported for these primary resurfacings and classified as acetabular, femoral, or both acetabular and femoral revisions. The survivorship of the different types of revisions was estimated using the Kaplan-Meier method and compared using proportional hazard models. Additionally, the outcome of a femoral-only revision was compared to that of primary conventional total hip arthroplasty. Acetabular-only revision had a high risk of re-revision compared to femoral-only and both acetabular and femoral revision (5-year cumulative per cent revision of 20%, 7%, and 5% respectively). Femoral-only revision had a risk of re-revision similar to that of revision of both the acetabular and femoral components. Femoral-only revision had over twice the risk of revision of primary conventional total hip arthroplasty. Revision of a primary resurfacing arthroplasty is associated with a major risk of re-revision. The best outcome is achieved when either the femoral-only or both the acetabular and femoral components are revised. Technically straightforward femoral-only revisions generally have a worse outcome than a primary conventional total hip arthroplasty.


The effects of commonly prescribed premolar extraction sequences on the curvature of the upper and lower lips.
The potential for premolar extractions to produce adverse facial effects after orthodontic treatment is still controversial. Detailed documentation of the predictability, or otherwise, of various soft tissue treatment effects would obviously be of assistance to clinical orthodontists in day-to-day treatment planning, by potentially refining the criteria for appropriate selection of various premolar extraction sequences. With this in mind, a retrospective lateral cephalometric study of 80 premolar extraction cases was undertaken to assess whether different patterns of premolar extraction do in fact produce predictably different lateral profile effects. A comparison was made of the changes in lip curvature after the extractions of all first premolars (4/4), all second premolars (5/5), or upper first and lower second premolars (4/5). Changes in the depths of curvature of both the upper and lower lips were not solely dependent on the selection of a particular premolar extraction sequence. Instead, there were wide ranges of individual variation in the changes in the depths of the lip curves. Therefore, in addition to the inherent soft tissue morphology of the lips in individual patients, it is the combined effect of the lip response to various dental and skeletal changes and the competent clinical management of extraction spaces that apparently affects the shapes of the lips within the lateral profile during treatment. In other words, it would seem possible for the clinician to carefully manage either first or second premolar extraction spaces while still protecting the facial profile.


Decreased insulin secretion in pregnant rats fed a low protein diet.
Low protein (LP) diet during pregnancy leads to reduced plasma insulin levels in rodents, but the underlying mechanisms remain unclear. Glucose is the primary insulin secretagogue, and enhanced glucose-stimulated insulin secretion (GSIS) in beta cells contributes to compensation for insulin resistance and maintenance of glucose homeostasis during pregnancy. In this study, we hypothesized that plasma insulin levels in pregnant rats fed LP diet are reduced due to disrupted GSIS of pancreatic islets. We first confirmed reduced plasma insulin levels, then investigated in vivo insulin secretion by glucose tolerance test and ex vivo GSIS of pancreatic islets in the presence of glucose at different doses, and KCl, glibenclamide, and L-arginine. Main findings include (1) plasma insulin levels were unaltered on day 10, but significantly reduced on days 14-22 of pregnancy in rats fed LP diet compared to those of control (CT) rats; (2) insulin sensitivity was unchanged, but glucose intolerance was more severe in pregnant rats fed LP diet; (3) GSIS in pancreatic islets was lower in LP rats compared to CT rats in the presence of glucose, KCl, and glibenclamide, and the response to L-arginine was abolished in LP rats; and (4) the total insulin content in pancreatic islets and expression of Ins2 were reduced in LP rats, but expression of Gcg was unaltered. These studies demonstrate that decreased GSIS in beta cells of LP rats contributes to reduced plasma insulin levels, which may lead to placental and fetal growth restriction and programs hypertension and other metabolic diseases in offspring.


Differential activation of IGF-II promoters P3 and P4 in Caco-2 cells during growth and differentiation.
Insulin-like growth factor (IGF)-II gene is overexpressed in colon cancers. Transcriptional up-regulation may be the major mechanism contributing to its overexpression. IGF-II messenger RNA (mRNA) levels are up-regulated during proliferation followed by a significant decline during differentiation of Caco-2 cells. Mechanisms underlying transcriptional regulation of the IGF-II gene promoters (P1-P4) have yet to be examined in colon cancers, which was the basis for this study. Ribonuclease protection assay was used to measure IGF-II mRNA derived from P1-P4. To determine if changes in the IGF-II transcripts reflected differences in promoter activity, transient transfection assays with the full-length P1-P4-luciferase expression vectors were performed. Both P3- and P4-derived transcripts were significantly up-regulated during the proliferative phase of the cells (days 3-6 in culture) and declined rapidly in cells undergoing differentiation (days 7-10); conversely, P1- and P2-derived transcripts were not detected. Similarly, transcriptional activity of P3 and P4 promoters reached peak levels by days 4-6 and declined rapidly thereafter. P1 and P2 were relatively inactive on all days. The activity of the P3 and P4 promoters may play a selective role in regulating IGF-II mRNA levels during growth and differentiation of colon cancer cells.


Tocopherol transport in the rat erythrocyte.
The transport of vitamin E (alpha-tocopherol) has been studied in the rat erythrocyte in vivo and in vitro. Uptake and efflux are independent of energy, but sensitive to temperature. Tocopherol is localized to the cell membrane. Rapid exchange takes place between erythrocytes and serum with an hourly fractional tocopherol efflux of 26%. The vitamin is transferred from the erythrocyte to the low density lipoproteins. These experiments indicate that tocopherol, like cholesterol, is a constituent of the erythrocyte membrane which is in dynamic equilibrium with the corresponding plasma compound.


Adjunctive Systemic Corticosteroids for Pulmonary Exacerbations of Cystic Fibrosis.
Pulmonary exacerbations (PEx) remain the most common cause of morbidity, recurrent hospitalization and diminished survival in people with CF (PWCF), and are characterized by excess inflammation. Corticosteroids are potent, widely available anti-inflammatory drugs. However, corticosteroid efficacy data from randomized controlled trials (RCTs) in PWCF are limited. To determine whether adjunctive systemic corticosteroid therapy is associated with improved outcomes in acute CF PEx. We performed a secondary analysis of STOP2, a large multicenter RCT of antimicrobial treatment durations for adult PWCF presenting with PEx, that included the use of corticosteroids as a stratification criterion in its randomization protocol. Corticosteroid treatment effects were determined after propensity score-matching for covariates including age, sex, baseline FEV1, genotype and randomization arm. The primary outcome measure was the change in percent predicted FEV1 (ppFEV1). Symptoms, time to next PEx and the incidence of adverse events (AEs) and serious adverse events (SAEs) were assessed as secondary endpoints. Phenotypic factors associated with the clinical decision to prescribe steroids were also investigated. Corticosteroids were prescribed for 168 of 982 PEx events in STOP2 (17%). Steroid prescription was associated with decreased baseline ppFEV1, increased age, and female sex. Co-treatment with corticosteroids was independent of treatment arm allocation, and did not result in greater mean ppFEV1 response, longer median time to next PEx or more substantial symptomatic improvement compared to propensity-matched PWCF receiving antibiotics alone. AEs were not increased in corticosteroid-treated PWCF. The total number of SAEs - but not the number of corticosteroid-related or PEx-ralated SAEs - was higher among patients receiving corticosteroids. Empiric, physician-directed treatment with systemic corticosteroids, while common, is not associated with improved clinical outcomes in PWCF receiving antibiotics for PEx.


Effect of sodium bisulfate on ammonia concentration, fly population, and manure pH in a horse barn.
To evaluate the effectiveness of altering the pH of manure with sodium bisulfate, thereby decreasing ammonia concentration and fly population in a horse barn environment. 4 mixed-breed pony stallions. The 4-week study was scheduled with 2 weeks of treatment (with 2 application rates) and 2 weeks with no treatment (control weeks). During treatment weeks, sodium bisulfate was applied daily to the top of the bedding and straw, then spread on top. Ponies were kept in the stalls 24 hours a day during the 7-day test period and stall cleaning was not done. On day 7 of each week, ammonia concentration, manure pH, number of flies on a fly tape, and fly-evasive behavior patterns were determined. Sodium bisulfate applied to the horse stall environment daily of either 2.3 or 4.5 kg/9.3 m2 (5 or 10lb/ 100 ft2) decreased ammonia concentration, manure pH, and number of flies in the stall environment, compared with a control period with no sodium bisulfate. Fly-evasive behavior patterns of ponies occupying the stalls, including tail swishes, head tosses, and kicks/strikes, were decreased during the period of sodium bisulfate application. Sodium bisulfate may be an effective method of decreasing ammonia concentration and may serve as a method of fly control in horse barns.


Pneumonia risk stratification in tropical Australia: does the SMART-COP score apply?
To examine the performance in tropical northern Australia of SMART-COP, a simple scoring system developed in temperate Australia to predict the need for intensive respiratory or vasopressor support (IRVS) in pneumonia patients. A prospective observational study of patients admitted to Royal Darwin Hospital in the Northern Territory with sepsis between August 2007 and May 2008. Chest x-rays were reviewed to confirm pneumonia, and each patient's SMART-COP score was assessed against the need for IRVS. Of 206 patients presenting with radiologically confirmed pneumonia, 184 were eligible for inclusion. The mean age of patients was 50.1 years, 65% were Indigenous and 56% were men. Overall, 38 patients (21%) required IRVS, and 18 patients (10%) died by Day 30. A SMART-COP score of >or= 3 had a sensitivity of only 71% for predicting the need for IRVS and 67% for 30-day mortality. As the variables most strongly associated with IRVS were serum albumin level < 35 g/L (odds ratio, 6.8) and Indigenous status (odds ratio, 2.3), we tested a modified scoring system (SMART-COP) that used a higher weighting for albumin and included Indigenous status. A SMART-COP score of >or= 3 had a sensitivity of 97% for IRVS and 100% for 30-day mortality. The SMART-COP score underestimates the severity of pneumonia in tropical northern Australia, but can be improved by using locally relevant additions.


Prompting clinicians about preventive care measures: a systematic review of randomized controlled trials.
Preventive care measures remain underutilized despite recommendations to increase their use. The objective of this review was to examine the characteristics, types, and effects of paper- and computer-based interventions for preventive care measures. The study provides an update to a previous systematic review. We included randomized controlled trials that implemented a physician reminder and measured the effects on the frequency of providing preventive care. Of the 1,535 articles identified, 28 met inclusion criteria and were combined with the 33 studies from the previous review. The studies involved 264 preventive care interventions, 4,638 clinicians and 144,605 patients. Implementation strategies included combined paper-based with computer generated reminders in 34 studies (56%), paper-based reminders in 19 studies (31%), and fully computerized reminders in 8 studies (13%). The average increase for the three strategies in delivering preventive care measures ranged between 12% and 14%. Cardiac care and smoking cessation reminders were most effective. Computer-generated prompts were the most commonly implemented reminders. Clinician reminders are a successful approach for increasing the rates of delivering preventive care; however, their effectiveness remains modest. Despite increased implementation of electronic health records, randomized controlled trials evaluating computerized reminder systems are infrequent.


No association between endogenous retinoic acid and human papillomavirus clearance or incident cervical lesions in Brazilian women.
Although oncogenic human papillomavirus (HPV) infections have been established as the necessary cause of cervical cancer, most HPV infections are transient and rarely progress to squamous cervical lesions. The activity of HPV is tightly associated with epithelial cell differentiation; therefore, regulators of differentiation, such as retinoic acid (RA), have been considered targets for the prevention of HPV-associated squamous intraepithelial lesion (SIL) development. The purpose of this study was to determine the association between circulating RA and early events in cervical carcinogenesis, specifically type-specific HPV clearance and SIL detection. Archived blood samples from 643 women participating in the Ludwig-McGill Cohort in São Paulo, Brazil, were analyzed by high-pressure liquid chromatography for three RA isomers (all-trans, 13-cis, and 9-cis-RA). A type-specific HPV clearance event was defined as two consecutive visits negative for an HPV type during follow-up for 364 HPV-positive women. Among the 643 women in this analysis, 78 were diagnosed with incident SIL. The probability of clearing an oncogenic HPV infection was not significantly different across RA isomer quartiles. There was a suggestion that increasing all-trans-RA increased the rate of nononcogenic HPV clearance (P-trend = 0.05). There was no association observed between serum RA levels and incident SIL. Our results suggest that elevated circulating RA isomer levels do not increase the rate of HPV clearance or reduce the risk of incident SIL. The role of RA in the inhibition of HPV-induced carcinogenesis, as shown in vitro, lacks confirmatory evidence within epidemiologic studies among women.


[Quantitative analysis of cell density and pattern of chondrocytes in transitory hyaline cartilage anlagen of human tarsal bones].
Changes in the distribution patterns of chondrocytes in the embryonic transitory hyaline cartilage of the tarsus were studied using 56 sagittal section of tarsi from 28 human embryos, with a vertex-foot length from 7.0 to 44.0 cm. The os naviculare and os cuboideum were not sliced in all sections. A total of 93,931 cells were transferred via a mirror to paper, and the resulting patterns were analyzed. Determinations included the numerical surface and volume density of the chondrocytes and the cell distribution patterns using the dispersion index. In the case of the calcaneus, talus, os naviculare and os cuboideum, the number of sliced cells found over a constant area decreases at a continuous linear rate in the course of the embryonic period. In all tarsal primordia the distribution pattern developed from regular to random. Particular conditions prevail immediately below the perichondrium and near the ossification centers which can be found in the calcaneus from the 6th month onwards.


Immunological and para-immunological mechanisms in spontaneous abortion: recent insights and future directions.
Unexplained spontaneous abortion in humans is currently thought to be related to genetic or immunologic factors. Animal models of spontaneous abortion have been used to try to gain some insight into the problems in humans. From studies of animal models there is increasing evidence that a variety of mechanisms may initiate pregnancy failure and participation of immunologic effectors may be a secondary event. Recent studies indicate that murine abortion that occurs spontaneously (without deliberate immunization by the investigator) may be initiated by para-immunologic host effector cells such as NK cells, macrophages and their toxins that possess selective toxicity which is not antigen-specific. The potential importance of local non-specific suppressive mechanisms in the uterus is also highlighted by their ability to block non-specific para-immune effector cell activation, and by their potential relationship to growth factors in decidual supernatants that promote placental cell growth in vitro. Levels of non-specific suppression do not appear low when fetal death can be attributed to a genetic mutation. The concept that different tissue components of the feto-placental unit serve as targets of different immunologic and para-immunologic effectors is defined. The new insights into the pathophysiology of murine abortion and its correction can provide a useful framework for further analysis of the role of para-immunologic effector mechanisms in unexplained abortion in humans and in non-murine veterinary conditions.


How We May Become Detached from Our Patients and What We Can Do If This Happens.
When clinicians provide clinical care or participate in ethics consultations, they may feel exceptionally painful emotions. When they do, they may distance themselves emotionally from patients and families. This distancing may harm these parties profoundly. It is therefore critical that clinicians avoid this distancing. In this piece, I present an approach that lies outside traditional practice that clinicians may use to try to avoid and even reverse this distancing, if and when they sense that this may be occurring. This approach may also benefit patients and families. It may increase their sense that their clinicians are working with them as allies to achieve their shared medical goals.


Inflammation, Biomarkers and Immuno-Oncology Pathways in Pancreatic Cancer.
It is estimated that pancreatic cancer will be the second leading cause of cancer-related deaths globally by 2030, highlighting the ongoing lack of effective treatment options for this devastating condition. There is a lack of reliable prognostic or predictive markers in pancreatic cancer to guide management decisions, whether for systemic chemotherapy, molecularly targeted therapies, or immunotherapies. To date, the results for targeted agents and immunotherapies in unselected populations of chemo-refractory pancreatic cancer have not met expectations. The reasons for this lack of efficacy of immunotherapy in pancreatic cancer are not completely understood. The challenges in pancreatic cancer include the physical barrier created by the dense desmoplastic stroma surrounding the tumor, chemokine-mediated exclusion of T cells, relatively poorer antigenicity compared to other solid tumors, paucity of infiltrating T cells within the tumor, ultimately leading to an immunosuppressive microenvironment. A better understanding of the role of inflammation in pancreatic cancer, its tumor microenvironment and individualized patient-related features, be they molecular, clinical or histopathological, would enable a more effective tailored approach to the management of pancreatic cancer. In this review, the role of inflammation, the immune tumor microenvironment and potential immune biomarkers in pancreatic cancer are explored.


A 6-month, randomized, double-blind, placebo-controlled pilot discontinuation trial following response to haloperidol treatment of psychosis and agitation in Alzheimer's disease.
In patients with Alzheimer's disease (AD) with psychosis or agitation that respond to haloperidol treatment, to evaluate the risk of relapse following discontinuation. In outpatients with AD with symptoms of psychosis or agitation, responders to 20 weeks of haloperidol (0.5-5 mg daily) were randomized to a 24-week, double-blind pilot trial of discontinuation on placebo versus continuation haloperidol. Phase A response criteria were minimum 50% reduction in three target symptoms, and improvement on the Clinical Global Impression-Change (CGI-C) score for psychosis/agitation. Phase B relapse criteria required 50% worsening in target symptoms and on the CGI-C. α = 0.1 was the significance criterion in this pilot study. Of 44 patients, 22 patients responded in Phase A. The sum score of target symptoms, and Brief Psychiatric Rating Scale (BPRS) psychosis and hostile suspiciousness factor scores, decreased in Phase A (p's < 0.001). Extrapyramidal signs increased in Phase A (p < 0.01). Of 22 responders, 21 patients entered Phase B, and 20 had at least one follow-up visit. Four of 10 patients (40%) on continuation haloperidol relapsed compared to eight of 10 patients on placebo (80%, χ(2) = 3.3, p = 0.07). In survival analyses, time to relapse was shorter on placebo than haloperidol (χ(2) = 4.1, p = 0.04). Haloperidol open treatment was efficacious, and relapse was greater on placebo than with haloperidol continuation. In patients with AD who have psychosis or agitation and respond to antipsychotic medication, the increased risk of relapse after discontinuation needs to be weighed against the side effects associated with continuing the medication.


HDAC6 inhibition promotes α-tubulin acetylation and ameliorates CMT2A peripheral neuropathy in mice.
Charcot-Marie-Tooth type 2A (CMT2A) peripheral neuropathy, the most common axonal form of CMT, is caused by dominantly inherited point mutations in the Mitofusin 2 (Mfn2) gene. It is characterized by progressive length-dependent degeneration of motor and sensory nerves with corresponding clinical features of motor and sensory impairment. There is no cure for CMT, and therapeutic approaches are limited to physical therapy, orthopedic devices, surgery, and analgesics. In this study we focus on histone deacetylase 6 (HDAC6) as a therapeutic target in a mouse model of mutant MFN2 (MFN2R94Q)-induced CMT2A. We report that these mice display progressive motor and sensory dysfunction as well as a significant decrease in α-tubulin acetylation in distal segments of long peripheral nerves. Treatment with a new, highly selective HDAC6 inhibitor, SW-100, was able to restore α-tubulin acetylation and ameliorate motor and sensory dysfunction when given either prior to or after the onset of symptoms. To confirm HDAC6 is the target for ameliorating the CMT2A phenotype, we show that genetic deletion of Hdac6 in CMT2A mice prevents the development of motor and sensory dysfunction. Our findings suggest α-tubulin acetylation defects in distal parts of nerves as a pathogenic mechanism and HDAC6 as a therapeutic target for CMT2A.


Effect of in ovo administration of an adult-derived microbiota on establishment of the intestinal microbiome in chickens.
OBJECTIVE To determine effects of in ovo administration of a probiotic on development of the intestinal microbiota of 2 genetic lineages (modern and heritage) of chickens. SAMPLE 10 newly hatched chicks and 40 fertile eggs to determine intestinal microbiota at hatch, 900 fertile eggs to determine effects of probiotic on hatchability, and 1,560 chicks from treated or control eggs. PROCEDURES A probiotic competitive-exclusion product derived from adult microbiota was administered in ovo to fertile eggs of both genetic lineages. Cecal contents and tissues were collected from embryos, newly hatched chicks, and chicks. A PCR assay was used to detect bacteria present within the cecum of newly hatched chicks. Fluorescence in situ hybridization and vitality staining were used to detect viable bacteria within intestines of embryos. The intestinal microbiota was assessed by use of 16S pyrosequencing. RESULTS Microscopic evaluation of embryonic cecal contents and tissues subjected to differential staining techniques revealed viable bacteria in low numbers. Development of the intestinal microbiota of broiler chicks of both genetic lineages was enhanced by in ovo administration of adult microbiota. Although the treatment increased diversity and affected composition of the microbiota of chicks, most bacterial species present in the probiotic were transient colonizers. However, the treatment decreased the abundance of undesirable bacterial species within heritage lineage chicks. CONCLUSIONS AND CLINICAL RELEVANCE In ovo inoculation of a probiotic competitive-exclusion product derived from adult microbiota may be a viable method of managing development of the microbiota and reducing the prevalence of pathogenic bacteria in chickens.