Ground-Truthing the Experiences of Homeless Older Adults' Recent Stays with Family and Friends: A Case Study of Participatory Data Analysis.
Many older homeless adults have recently stayed with family or friends, yet little is understood about these experiences. We conducted a multimethod qualitative study of older unhoused adults in Oakland, California. First, we conducted in-depth interviews among older adults experiencing homelessness with recent stays with a housed family member or friend (n=46), hosts (n=19), and program key informants (n=11). Second, we developed thematic summaries in the form of character-based composite stories, which were presented to a Council of Elders with lived experiences of homelessness, to explore reactions, referred to as ground-truthing, a form of participatory data analysis. Predominantly, participants were African American men. Barriers included structural factors (discrimination), policy (lease restrictions), community (violence), interpersonal factors (power dynamics), and individual factors (health problems). Factors enhancing stays included inter-generational support and leveraging resources. Ground-truthing discussions reinforced and expanded upon findings (e.g., importance of neighborhood identity, training needs, how self-improvement affects readiness to live with others).


Litter quality and the law of the most limiting: Opportunities for restoring nutrient cycles in acidified forest soils.
The adverse effects of soil acidification are extensive and may result in hampered ecosystem functioning. Admixture of tree species with nutrient rich litter has been proposed to restore acidified forest soils and improve forest vitality, productivity and resilience. However, it is common belief that litter effects are insufficiently functional for restoration of poorly buffered sandy soils. Therefore we examined the effect of leaf litter on the forest floor, soil chemistry and soil biota in temperate forest stands along a range of sandy soil types in Belgium, the Netherlands and Germany. Specifically, we address: i) Which tree litter properties contribute most to the mitigation of soil acidification effects and ii) Do rich litter species have the potential to improve the belowground nutrient status of poorly buffered, sandy soils? Our analysis using structural equation modelling shows that litter base cation concentration is the decisive trait for the dominating soil buffering mechanism in forests that are heavily influenced by atmospheric nitrogen (N) deposition. This is in contrast with studies in which leaf litter quality is summarized by C/N ratio. We suggest that the concept of rich litter is context dependent and should consider Liebig's law of the most limiting: if N is not limiting in the ecosystem, litter C/N becomes of low importance, while base cations (calcium, magnesium, potassium) become determining. We further find that on poorly buffered soils, tree species with rich litter induce fast nutrient cycling, sustain higher earthworm biomass and keep topsoil base saturation above a threshold of 30%. Hence, rich litter can trigger a regime shift to the exchange buffer domain in sandy soils. This highlights that admixing tree species with litter rich in base cations is a promising measure to remediate soil properties on acidified sandy soils that receive, or have received, high inputs of N via deposition.


A morbidity score for congenital heart surgery based on observed complications.
The aim of this study was to develop a morbidity score based on observed postoperative complications after congenital heart surgery. Conditions or diseases that impair patients after congenital heart surgery or increase costs of hospital stay were called complications and attributed to scores ranging from 1 (mild) to 4 (severe) points, according to estimated severity or costliness. 'No complication' was assigned 0.5 points. From January to March 2011, scores for each observed 'complication' for every main (primary) surgical procedure were recorded and a morbidity score was calculated. In conformity with the Aristotle score methodology, if the sum of observed complication scores amounted to more than 5 points, a morbidity score of only 5 points was attributed. The estimated morbidity score was compared with the morbidity score attributed by the Aristotle basic complexity (ABC) score. One hundred and thirty-nine primary procedures were carried out. The mean ABC and Aristotle comprehensive complexity scores reached 8.31 ± 2.52 and 9.62 ± 3.47 points, respectively. Two patients died. No complication was detected after 46 procedures. Overall, there were 185 listed 'morbidity' conditions in connection with the other 93 surgical procedures, rendering a total score of 385 points. The most frequent event was 'mechanical ventilation 25-95 h': n = 39. The mean morbidity score was 2.14 ± 1.63. The morbidity scores ranged from 0.5 points (n = 46) to 5 points (n = 23) with a median of 2.0 points. The scores for 11 different procedures that were performed at least five times positively correlated with the corresponding Aristotle morbidity scores: Pearson's coefficient r = 0.75. But the morbidity score for bidirectional cavopulmonary anastomosis (3.14 ± 1.77) was higher than the corresponding Aristotle morbidity score (2.0). It was lower for 'conduit placement, right ventricle to pulmonary artery': 1.08 ± 0.97, versus 2.0, and for arterial switch operation: 2.08 ± 1.11, versus 3.0. The reported morbidity scores need to be tested on larger series and in different institutions. The introduced morbidity score has the potential to quantify postoperative complications accurately. Its estimation over time can facilitate the assessment of quality of congenital heart surgery. It will allow comparison of morbidity outcomes across institutions with different case-mixes.


[Formation of endogenous pyrogen by mononuclear phagocytes].
Production of endogenous pyrogen by human and rabbit blood monocytes in response to stimulation with agents of different origin was studied by inhibitory analysis under comparable conditions. Actinomycin D and cytochalasin B were applied. New evidence was obtained about an important role in the mechanism of activation of mononuclear phagocytes of initial interaction between a stimulating agent and the leukocyte membrane and of the biphasic process of endogenous pyrogen production.


Development of monoclonal antibodies against creatine kinase MB2.
An increase of creatine kinase MB (CKMB) in serum has long been used as a marker for acute myocardial infarction (AMI). It is important with an early diagnosis and since the amount of the CKMB2 isoform rises above reference levels much earlier than the total creatine kinase amount, quantification of CKMB isoforms could be a feasible alternative for early analysis. The two CKMB isoforms differ by only one C-terminal lysine residue, which makes it difficult to separate one from the other. To overcome this problem, monoclonal antibodies were produced using unique peptides as antigen in hybridoma technology. Two peptides with 16 and 15 amino acids corresponding to the C-terminal end of the M-subunits of CKMB2 and CKMB1, respectively, were conjugated to keyhole limpet hemocyanin and used as antigens. Sixteen different monoclonal antibodies to these peptides were obtained and characterized. Their specificity was analyzed by immunoassay and 10 of the antibodies showed cross-reactive binding to creatine kinase. Surface plasmon resonance based biosensor analysis was used to determine affinity and kinetics towards the peptides and the epitopes of four of the antibodies were studied by means of phage display. Some of these antibodies have binding properties that might qualify them for use in the establishment of procedures allowing early diagnosis of AMI.


Heat stress, plasma concentrations of adrenaline, noradrenaline, 5-hydroxytryptamine and cortisol, mood state and cognitive performance.
The primary aims of this paper were to examine the effect of heat stress on working memory, choice reaction time and mood state, and to investigate the relationship between heat induced changes in plasma concentrations of selected neurotransmitters and hormones, and cognition. Heat stress resulted in a deterioration of performance on a central executive task (random movement generation) but not on verbal and spatial recall, and choice reaction time tasks. Perceptions of vigour decreased and fatigue increased following exposure to heat stress. Plasma concentrations of cortisol and 5-hydroxytryptamine significantly increased following exposure to heat. Regression analyses showed that percent body mass loss and change from baseline (Delta) concentrations of cortisol, post-exposure to heat, were significant predictors of Delta random movement generation and Delta fatigue. A secondary purpose was to examine the effect of recovery on cognition and mood. Following recovery, the performance of the central executive task was poorer than pre-treatment. Mood states, catecholamines and 5-hydroxytryptamine concentrations returned to pre-treatment values, but cortisol fell to a level significantly lower. Regression correlations showed that Delta adrenaline and Delta scores, post-recovery, on the central executive task were significantly correlated. Delta noradrenaline correlated significantly with Delta fatigue. It was concluded that heat stress results in deterioration in the performance of central executive tasks and perceptions of mood state, and that this can be predicted by changes in body mass loss and plasma concentrations of the hormones cortisol and adrenaline.


Distinct trafficking and localization of STEVOR proteins in three stages of the Plasmodium falciparum life cycle.
The genome of Plasmodium falciparum harbors three extensive multigene families, var, rif, and stevor (for subtelomeric variable open reading frame), located mainly in the subtelomeric regions of the parasite's 14 chromosomes. STEVOR variants are known to be expressed in asexual parasites, but no function has as yet been ascribed to this protein family. We have examined the expression of STEVOR proteins in intraerythrocytic sexual stages, gametocytes, and extracellular sporozoites isolated from infected Anopheles mosquitoes. In gametocytes, stevor transcripts appear transiently early in development but STEVOR proteins persist for several days and are transported out of the parasite, travel through the host cell cytoplasm, and localize to the erythrocyte plasma membrane. In contrast to asexual parasites, gametocytes move STEVOR to the periphery via a trafficking pathway independent of Maurer's clefts. In sporozoites, STEVOR appear dispersed throughout the cytoplasm in vesicle-like structures. The pattern of STEVOR localization we have observed in gametocytes and sporozoites differs significantly from that in asexual parasite stages. STEVOR variants are therefore likely to perform different functions in each stage of the parasites life cycle in which they occur.


Tailoring magnetic dipole emission with plasmonic split-ring resonators.
We numerically explore the emission behavior of magnetic dipole emitters located next to resonant plasmonic split-ring resonators (SRRs), which are well known for their large magnetic moment at their fundamental resonance in the near infrared. Our results are compared to the situation for electric dipole emitters, where the SRR can be described by solely its electric dipole moment. We show that a similar approach in the case of magnetic dipole emitters is not sufficient, as the symmetry breaking due to the gap has to be taken into account. We demonstrate how retardation between the emitter and the SRR can be used as an additional degree of freedom to manipulate the emission spectrum. Our concept will pave the road towards efficient plasmonic antennas for magnetic dipole emitters.


Vagal innervation modulates motor pattern but not initiation of canine gastric migrating motor complex.
To determine the role of vagal nerves in initiation and modulation of the gastric migrating motor complex (MMC), motor activity was recorded in four dogs before and after total abdominal vagotomy during fasting, after exogenous intravenous motilin and insulin, and after feeding. After vagotomy, a temporally coordinated cyclic gastric and small bowel MMC persisted with an unchanged period. During gastric phase III, vagotomy decreased number of contractions (42 +/- 4 vs. 16 +/- 2), number of groupings of contractions (14 +/- 1 vs. 7 +/- 1), and motility index (12 +/- 1 vs. 10 +/- 1) and increased the duration between groupings (1 +/- 1 vs. 3 +/- 1 min) (P < 0.05 in each). Before and after vagotomy, motilin and insulin induced a premature MMC with minor changes in contractile pattern. A 200-g liver meal but not a 50-g liver meal inhibited the gastric MMC after vagotomy. A cyclic MMC persisted after vagotomy, but the contractile pattern during gastric phase III was altered. After a short recovery period, vagal innervation to the stomach modulates the pattern but not the presence of gastric interdigestive motility during phase III.


Clinical and SEM characterization of prolonged retention of a primary tooth with pulpectomy.
Root canal filling with zinc oxide-eugenol (ZOE) paste following primary tooth pulpectomy is a common practice in pediatric dentistry. This material offers high clinical and radiographic success rates. In some cases, however, it is not resorbed along with the root of the primary tooth. The aim of this study was to describe a case of prolonged retention of a primary maxillary incisor that was subjected to pulpectomy and filled with ZOE paste in order to characterize the aspects of root resorption using scanning electron microscopy.


Automatic 300-4 K temperature cycling apparatus.
The apparatus described here automatically cycles small samples between 300 and 4 K by alternately raising and lowering the sample through the neck of a commercial liquid helium storage Dewar. A bellows, which is pressurized by the helium boil-off gas, provides all of the required mechanical motion. By utilizing the cooling available from the boil-off gas, liquid helium helium consumption is limited to 0.03 l/cyc for a 12-g sample. Cycle times can be as short as 5 min.


Dermatitis artefacta.
Dermatitis artefacta, also known as factitious dermatitis, is a condition in which cutaneous lesions are self-inflicted and are the result or manifestation of some psychological conflicts. This report presents the case of a 20-year-old man, whose initial presentation resembled a dermatological disorder. Psychological and personality factors as well as issues in the management are discussed.


Isolation and genetic analysis of a mutation that suppresses the auxotrophies of superoxide dismutase-deficient Escherichia coli K12.
The most striking phenotype associated with superoxide dismutase (SOD) deficiency in Escherichia coli is the inability to grow in aerobic minimal medium, which is due to the sensitivity of several amino acid biosynthetic pathways to superoxide. We have isolated two classes of pseudorevertants that grow on minimal medium at modest rates. Of these, the class that exhibited the faster growth carries mutations at a single locus, denoted ssa, which was mapped to 4 min on the E. coli chromosome. This class constituted the majority of the spontaneous pseudorevertants that were selected by the transfer of independent SOD-deficient cultures in minimal medium from anaerobic to aerobic growth conditions. Pseudoreversion at ssa suppressed requirements for a variety of unrelated amino acid supplements. Further, the SOD-deficient strains were unable to assimilate diaminopimelic acid from the growth medium, whereas the ssa pseudorevertants did so. The viability of these pseudorevertants indicates that superoxide-sensitive biosynthetic enzymes do retain some function in SOD-deficient cells during aerobic growth.


Amplification of major histocompatibility complex class II gene diversity by intraexonic recombination.
The roles of mutational and recombinational processes in the diversification of the exon encoding the antigen binding site in the murine major histocompatibility complex class II gene Ab were assessed by phylogenetic analysis of allelic nucleotide sequences. A total of 46 alleles of Ab exon 2 from 12 Mus species or subspecies and 2 Rattus species were sequenced after amplification by the polymerase chain reaction. Reliable allelic genealogies could not be determined by phylogenetic analyses, due to extensive homoplasy in the data set. This homoplasy results from the shuffling of polymorphisms between alleles by recombinational processes, indicating that polymorphisms in the antigen binding site encoded by Ab are generated by a combination of two processes. First, the accumulation of point mutations has produced highly divergent polymorphic sequence motifs in five regions of Ab exon 2, each encoding a portion of the binding site. Some of these motifs have persisted as polymorphisms in rodents since before the divergence of mouse and rat (greater than 10 million years ago). The second process mediating Ab diversification involves the shuffling of these polymorphic sequence motifs into numerous allelic combinations by repeated intraexonic recombination. Site-specific hyperrecombinational mechanisms are not involved in this process within the exon. We postulate that these mechanisms continuously generate new Ab alleles with highly divergent binding sites from which alleles with advantageous antigen-binding properties are selectively maintained by some form of balancing selection.


Distributing working memory resources during problem solving.
This study examines how problem solvers distribute working memory demands over internal and external resources. Participants recorded notes while performing an arithmetic task. They recorded a majority of intermediate results and labeled many of those results (e.g., "C = 10"). When more effort was required to take notes, participants recorded fewer results. Participants with a consistent goal structure recorded fewer results and with practice labeled fewer recorded results than those with varied goal structures. When notes were displayed in a consistent spatial arrangement participants labeled fewer recorded results than when notes appeared in varied locations. These findings indicate that individuals use explicit and implicit strategies for indexing intermediate results. The data support the view that individuals flexibly distribute working memory over internal and external resources in response to situational cost-benefit considerations.


A study on the impact of the number of family generations on intergenerational support for centenarians: a study in a Chinese 'longevity city'.
As a natural source of support for the elderly, the family is an important channel for achieving a sense of security, happiness, and worthiness in old age. In this study, we analysed the characteristics of intergenerational support in families of centenarians and explored the impact of the number of family generations on intergenerational support. We conducted a cross-sectional survey between April 2020 and January 2021 among 62 elderly people aged 99+ in Rugao, China, one of six 'longevity cities' in the world. Assisted by the researchers, centenarians completed questionnaires with details pertaining to general demographics, intergenerational support, and other aspects. We used a logistic regression model to analyse the influence of the number of family generations on intergenerational support that the centenarians received with respect to economic, living, and emotional aspects. Centenarians were primarily recipients of care in their families, and received intergenerational support mainly for their declined physical functions and limited self-care ability. The study results revealed that the greater the number of generations comprising the family, the greater was the intergenerational life care and emotional comfort provided for centenarians by the family. In this study, we found a positive effect of the number of family generations on intergenerational support for centenarians. The government and society should promote the tradition of respecting, caring for, and honouring the elderly while paying close attention to the dynamic changes in the family structure of centenarians in promoting high-quality and sustainable development of the people, economy, and society.


Identification of adult stem cells in Schwalbe's line region of the primate eye.
To identify stem cells in the chamber angle of the monkey eye by detection of 5-bromo-2'-deoxyuridine (BrdU) long-term retention. Four cynomolgus monkeys were treated with BrdU via subcutaneous pumps for 4 weeks. The eyes of two animals were processed immediately thereafter (group 1) while in the other animals, BrdU treatment was discontinued for 4 weeks to allow identification of cells with long-term BrdU retention (group 2). The number of BrdU-positive nuclei was quantified, and the cells were characterized by immunohistochemistry and transmission electron microscopy (TEM). The number of BrdU-positive cells was higher at Schwalbe's line covering the peripheral end of Descemet's membrane than in Schlemm's canal (SC) endothelium, trabecular meshwork (TM), and scleral spur (SS). Labeling with BrdU in SC, TM, and SS was less intense and the number of labeled cells was smaller in group 2 than in group 1. In contrast, in cells of Schwalbe's line the intensity of BrdU staining and the number of BrdU-positive cells was similar when group 1 and 2 monkeys were compared with each other, indicating long-term BrdU retention. Cells that were BrdU-positive in Schwalbe's line region stained for the stem cell marker OCT4. Details of a stem cell niche in Schwalbe's line region were identified by TEM. We provide evidence for a niche in the Schwalbe's line region harboring cells with long-term BrdU retention and OCT4 immunoreactivity. The cells likely constitute a population of adult stem cells with the capability to compensate for the loss of TM and/or corneal endothelial cells.


Effects of melatonin administration to cashmere goats on cashmere production and hair follicle characteristics in two consecutive cashmere growth cycles.
The objective of the study was to determine the long-term effects of melatonin treatment on cashmere production and hair follicle populations in cashmere goats and their activity in two consecutive cashmere growth cycles. Twenty-four female Inner Mongolian Cashmere goats were randomly allocated to two groups (n = 12), one of which received melatonin implants, the other being an untreated control group. Melatonin implants were subcutaneously inserted behind the ear at a dose of 2 mg/kg live weight on two occasions -April 30 and June 30, 2016. Hair samples were collected by combing in April of 2017 and 2018, and the weight, staple length, and diameter of the cashmere fibers were measured. Blood and skin samples were collected monthly between April and September 2016, and in April and September in 2017 for the analysis of melatonin concentration and the characteristics of secondary hair follicle populations, respectively. The results indicated that serum melatonin concentration in the treated goats was elevated (P < 0.05) relative to that of the control group, but only during the first growth cycle. Melatonin treatment of cashmere goats in one cashmere growth cycle increased the weight, staple length, and density (all P < 0.05) of the cashmere fibers and decreased fiber diameter (P < 0.01), but did not affect the characteristics of cashmere production in the subsequent annual cycle. Melatonin treatment had no effect on the population of skin secondary hair follicles for two consecutive cycles. However, in the first growth cycle after treatment, it advanced the onset of activity of skin secondary hair follicles by 2 mo (P < 0.05), and it increased the number of follicles that were active (P < 0.05). In summary, the melatonin treatment of cashmere goats in one cashmere growth cycle improved cashmere production for that cycle only, with no residual effects on the subsequent cycle, a technique acceptable to the cashmere goat industry. The enhancement of cashmere production after the treatment of goats with melatonin appears to involve the acceleration of the annual regeneration of skin secondary hair follicles and increased population of active secondary hair follicles in the skin of cashmere goats.


Pseudoleukemia. Ineffective regulation of granulopoiesis.
A neonate with Down's syndrome confirmed by chromosomal analysis had leukocytosis, with a WBC count of 48,000/cu mm, at age 13 days. Differential counts showed 70% to 90% blast forms by light microscopy. The blast-cell granules stained negatively with Sudan black B and alpha-napthyl butyrate esterase. Myeloperoxidase activity evaluated light and electron microscopy was negative. The granules stained positively with PAS, beta-glucuronidase, and tartrate-sensitive acid phosphatase. Indirect immunofluorescence for terminal-deoxynucleotidyl-transferase was negative. Our results show that the blast cells of pseudoleukemia may be of lymphoid origin.


Heterogeneity of ascorbate free radical reductase in the human lens.
The soluble ascorbate free radical (AFR) reductases in the human lens were separated into many isoforms in the range of pI 5-7 by native isoelectric focusing. In the two-dimensional gel electrophoresis, however, two main proteins with molecular weights of 20-25 kD were commonly identified to each isoform. The observed heterogeneity of the human lens AFR reductase is very similar to those reported for beta- and gamma-crystallins in aged and cataractous human lenses. From these results, it is suggested that some of the isoforms of the lens AFR reductase, especially the more acidic isoforms, may be formed by posttranslational modifications.


Untreated thyroid autoantibody-negative SCH increases the risk of spontaneous abortions.
Numerous studies have found that subclinical hypothyroidism (SCH) may increase adverse pregnancy outcomes; however, the benefit of levothyroxine (LT4) treatment remains controversial. The 2017 guidelines of the American Thyroid Association weakly recommended LT4 therapy for serum antithyroid peroxidase antibody (TPOAb)-negative women with thyroid-stimulating hormone (TSH) concentrations greater than the pregnancy-specific reference range and below 10.0 mU/L. Therefore, the primary goal of this study was to investigate the correlation between thyroid autoantibody-negative SCH with or without LT4 treatment and adverse pregnancy outcomes. We prospectively enrolled 1868 consecutive pregnant women. Finally, 1344 women were involved in the study according to the inclusion and exclusion criteria. Assays for TSH, free thyroxine (FT4), TPOAb, anti-thyroglobulin antibody, and laboratory indicators were performed. The participants were divided into the euthyroid (ET) group (n = 1250) and the SCH group(n = 94). The SCH group was further divided into LT4 group (n = 40) and non-LT4 group(n = 54). The laboratory indicators and pregnancy outcomes were evaluated during follow-ups. Maternal age, BMI, parity, and the history of spontaneous abortion did not differ significantly between the ET group and the different SCH groups. There were no significant differences in lipid profile and homocysteine levels between ET and SCH group in the first and third trimester of pregnancy. After adjusting the confounding factors, the non-LT4 group was a risk factor for spontaneous abortion (odds ratio: 3.141, 95% CI: 1.060-9.302). Survival analysis showed that the time of abortion was different between the ET group and SCH group (log-rank P= 0.042). The spontaneous abortion in SCH, especially in non-LT4, group mainly occurred in the first trimester of pregnancy. Thyroid autoantibody negative-SCH seems to be associated with increased risk of spontaneous abortions during the first trimester of pregnancy. LT4 therapy in this patient population might be beneficial to reduce adverse pregnancy outcomes.


[Effect of cortisol on glycosaminoglycan synthesis and growth of diploid, human fibroblasts (WI-38) in relation to in vitro aging].
Addition of cortisol (hydrocortisone; 1.4 X 10(-7) M) to the culture medium (BME with non-essential amino acids and 10% fetal calf serum) of human fibroblasts resulted in increased proliferative activity--in regard to cell number and incorporation rates of [3H]-thymidine into DNA and [14C]-leucine into protein--and in an increased saturation density. Glycosaminoglycan (GAG) synthesis rates (as measured by incorporation of [14C]-glucosamine or [35S]-sulfate) of the various GAG types secreted into the culture medium (hyaluronic acid, chondroitin sulfate, dermatan sulfate, heparan sulfate) were evenly inhibited by ca. 25% in the case of cortisol-treated Phase-II cultures. The dose-effect relationship revealed a median effective cortisol dose of ca. 0.01 microM for Phase-II cultures. Phase-III ("senescent") cultures revealed an elevated sensitivity as well in regard to cortisol concentration as to the extent of the inhibitory effect. Contrary to the medium GAGs, the pattern of the cell-bound GAGs was changed by cortisol (1.4 X 10(-7) M), with an increase of hyaluronic acid synthesis and a decrease of the sulfated GAGs. This cell-bound hyaluronic acid was predominantly removable by trypsin-treatment and therefore regarded to be localized at the cell surface or pericellularly. Also, following long-term cultivation (ca. 15 population-doublings) in the presence of cortisol, the synthesis of cell-bound hyaluronic acid was stimulated. Since cellular hyaluronic acid decreases during senescence of WI-38 cultures (Sluke et al., 1981), the cortisol effect in regard to GAG synthesis is in line with a "counter-aging" influence.(ABSTRACT TRUNCATED AT 250 WORDS)


Microwave Synthesis of Thionated Naphthalene Diimide-Based Small Molecules and Polymers.
Thionated naphthalene diimides (NDIs) are promising materials for n-type organic semiconductors; despite their potential, synthetic routes to thionated NDIs are generally lengthy, nonselective, and low yielding and their polymeric analogues have yet to be reported in the literature. Here, we describe the rapid and selective thionation of thiophene- and selenophene-flanked NDIs using microwave irradiation and excess Lawesson's reagent. Remarkably, >99% conversion to the trans-dithionated product is observed by NMR within 45 min. Steric effects imparted by NDI core substituents prevent excess thionation, simplifying purification procedures. We apply this methodology to the postpolymerization thionation of NDI-based polymers to afford a series of polymers with varying degrees of thionation. Thionated NDIs exhibit bathochromic shifts of up to ∼100 nm in localized absorption maxima and increased electron affinities.


Evolution of nutritional management of acute malnutrition.
Wasting, kwashiorkor and stunting are not usually due to either protein or energy deficiency. Treatment based upon this concept results in high mortality rates, and failure of treated children to return physiologically to normal. They become relatively obese with insufficient lean tissue. Preventive strategies have also failed. Wasting and stunting are primarily due to deficiency of type II nutrients and kwashiorkor probably due to deficiency of several type I nutrients that confer resistance to oxidative stress. Modern dietary treatments are based upon the F75 formula whilst the child is sick without an appetite, followed by F100 for rapid gain of weight. Derivative, ready-to-use therapeutic foods (RUTF) allow treatment of large numbers of children at home, are preferred by mothers and dramatically improve coverage. Children are indentified by screening in the community and treated before complications arise, using simple protocols. Successful treatment of the sick children with severe malnutrition not only depends upon these products, but appropriate management of complications. The physiology of the malnourished child is completely different from the normal child and many drugs and treatments that are safe in children with normal physiology are fatal for the malnourished child. In particular, the diagnosis and management of diarrhea and dehydration is different in the malnourished child. Giving standard treatment frequently leads to circulatory overload and death from heart failure. The challenge now is to find successful local ways to prevent malnutrition and achieve nutritional security. Until prevention works, we have to rely on fortified foods for treatment and convalescence from illness.


Expression of transforming growth factor-beta isoforms in human glomerular diseases.
Protein and mRNA expression of TGF-beta isoforms, TGF-beta 1, -beta 2 and -beta 3, and deposition of fibronectin containing extra domain A (fibronectin EDA+) and plasminogen activator inhibitor-1 (PAI-1) were studied in human chronic glomerulonephritis and diabetic nephropathy. Normal kidneys showed similar, weak immunostaining for all three TGF-beta isoforms. TGF-beta mRNA expression was weak for all isoforms with TGF-beta 1 > TGF-beta 3 >> TGF-beta 2. In thin basement membrane disease and minimal change disease, disorders where extracellular matrix accumulation is not a feature, immunoreactivity and mRNA expression did not differ from normal. In contrast, diseases characterized by extracellular matrix accumulation (IgA nephropathy, focal and segmental glomerulosclerosis, crescentic glomerulonephritis, lupus nephritis and diabetic nephropathy) all showed significantly increased expression of the three TGF-beta isoforms in glomeruli and the tubulointerstitium. While glomerular and tubulointerstitial deposition of two matrix components induced by TGF-beta, fibronectin EDA+ and PAI-1, was significantly elevated in all diseases with matrix accumulation, correlation analysis revealed a close relationship primarily with TGF-beta 1. We conclude that, for a spectrum of human glomerular disorders, increased protein expression of all three TGF-beta isoforms and proteins induced by TGF-beta is associated with pathological accumulation of extracellular matrix.


Ethanol increases nuclear factor-kappa B activity in human astroglial cells.
Alcohol abuse adversely affects essentially all the organs of the body, either directly or indirectly. Ethanol may contribute to brain damage via inflammation. Ethanol may also alter CNS immunocompetence and further the progression of certain CNS infections. Nuclear factor (NF)-kappa B helps regulate inflammatory gene expression in glia. It is possible that ethanol effects on CNS pathology are partly a consequence of ethanol modulation of NF-kappa B-associated pathways in glia. We have assessed the effects of 0.5-6 h ethanol exposure on cytokine (5 ng/ml interleukin-1 beta + 100 ng/ml interferon gamma + 30 ng/ml tumor necrosis factor-alpha)-induced NF-kappa B activation in human A172 astroglial cells. Immunoblot analysis indicated that NF-kappa B p65 nuclear translocation occurred within 0.5 h after cytokine stimulation. Stimulation in the presence of ethanol resulted in increased nuclear p65 levels at 3 h, with 200 mM causing a greater increase than 50 mM ethanol. Gel shift assay data suggested that cytokine-induced NF-kappa B binding activity was greatest in cells exposed to 50 mM ethanol, followed by 200 and 0 mM ethanol exposed cells, respectively. Thus, in cytokine-stimulated cells, 200 mM ethanol resulted in greater nuclear p65 levels, yet, 50 mM ethanol exposure resulted in more pronounced DNA binding by NF-kappa B. These findings demonstrate that acute ethanol enhances p65 activity in human astroglia and further support the hypothesis that ethanol-mediated brain pathology involves modulation of NF-kappa B pathways. A better understanding of the mechanistic events involved in ethanol-induced CNS pathology should provide for therapeutic strategies to combat detrimental effects of alcohol on the CNS.


Profilings of MicroRNAs in the Liver of Common Carp (Cyprinus carpio) Infected with Flavobacterium columnare.
MicroRNAs (miRNAs) play important roles in regulation of many biological processes in eukaryotes, including pathogen infection and host interactions. Flavobacterium columnare (FC) infection can cause great economic loss of common carp (Cyprinus carpio) which is one of the most important cultured fish in the world. However, miRNAs in response to FC infection in common carp has not been characterized. To identify specific miRNAs involved in common carp infected with FC, we performed microRNA sequencing using livers of common carp infected with and without FC. A total of 698 miRNAs were identified, including 142 which were identified and deposited in the miRbase database (Available online: http://www.mirbase.org/) and 556 had only predicted miRNAs. Among the deposited miRNAs, eight miRNAs were first identified in common carp. Thirty of the 698 miRNAs were differentially expressed miRNAs (DIE-miRNAs) between the FC infected and control samples. From the DIE-miRNAs, seven were selected randomly and their expression profiles were confirmed to be consistent with the microRNA sequencing results using RT-PCR and qRT-PCR. In addition, a total of 27,363 target genes of the 30 DIE-miRNAs were predicted. The target genes were enriched in five Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including focal adhesion, extracellular matrix (ECM)-receptor interaction, erythroblastic leukemia viral oncogene homolog (ErbB) signaling pathway, regulation of actin cytoskeleton, and adherent junction. The miRNA expression profile of the liver of common carp infected with FC will pave the way for the development of effective strategies to fight against FC infection.


Synthesis of low-shrinkage polymerizable liquid-crystal monomers.
Polymerization shrinkage remains a major barrier to the universal use of resin restorative in large posterior cavity preparations. A new bifunctional liquid crystal (LC) monomer, 2-(t-butyl), 1,4-bis-[4-(6-acryloxy-hexane-1-oxy)-benzoyloxy] benzene, with exceptionally low polymerization shrinkage, has recently been discovered. The purpose of this communication is to report a new, easy, high-yield synthetic route to synthesize this compound in comfortable larger batches. Synthetic and isolation details, chemical characteristics, and the polymerizable properties of a new structurally related by-product monomer, namely, 2-(t-butyl),1-[4-(6-acryloxy-hexane-1-oxy)-benzoyloxy], 4-[4-[6-(3-acryl oxy-propionoxy)-hexane-1-oxy]-benzoyloxy]-benzene, is also reported. The structural confirmation of this by-product indicates that it resulted from the Michael-type addition of acrylate ion on one of the terminal acryloxy groups of 2-(t-butyl), 1,4-bis-[4-(6-acryloxy-hexane-1-oxy)-benzoyloxy] benzene. The by-product itself, as well as the natural blend of the aforesaid both products as formed in the reaction mixture, also polymerized at room temperature with lesser volume shrinkage as compared to the conventional control (GTE) at similar degrees of conversion.


Observation of site-controlled localized charged excitons in CrI3/WSe2 heterostructures.
Isolated spins are the focus of intense scientific exploration due to their potential role as qubits for quantum information science. Optical access to single spins, demonstrated in III-V semiconducting quantum dots, has fueled research aimed at realizing quantum networks. More recently, quantum emitters in atomically thin materials such as tungsten diselenide have been demonstrated to host optically addressable single spins by means of electrostatic doping the localized excitons. Electrostatic doping is not the only route to charging localized quantum emitters and another path forward is through band structure engineering using van der Waals heterojunctions. Critical to this second approach is to interface tungsten diselenide with other van der Waals materials with relative band-alignments conducive to the phenomenon of charge transfer. In this work we show that the Type-II band-alignment between tungsten diselenide and chromium triiodide can be exploited to excite localized charged excitons in tungsten diselenide. Leveraging spin-dependent charge transfer in the device, we demonstrate spin selectivity in the preparation of the spin-valley state of localized single holes. Combined with the use of strain-inducing nanopillars to coordinate the spatial location of tungsten diselenide quantum emitters, we uncover the possibility of realizing large-scale deterministic arrays of optically addressable spin-valley holes in a solid state platform.


Occurrence and fate of pharmaceuticals and personal care products in drinking water in southern China.
Occurrence and fate of pharmaceuticals and personal care products (PPCPs) in drinking water was investigated in southern China. Fifteen and twelve PPCPs were detected with concentrations of 0-36 ng L(-1) in source water and of 0-20 ng L(-1) in treated water, respectively. Four PPCPs were detected with concentrations of approximately 1 ng L(-1) in drinking water of distribution network. Conventional water treatment processes removed the types and average concentrations of PPCPs by 30% and above 50%, respectively. Advanced water treatment processes were more efficient in the removal of most PPCPs, with the types and concentrations reduced by 50% and approximately 90%, respectively. Molecular properties of PPCPs had an important influence on their behaviors during water treatment. pK(a) (acidity coefficient) and K(oc) (organic carbon partition coefficient) of PPCPs appeared to have a combined effect on PPCPs removal during coagulation and oxidation. Adsorption and biodegradation were two possible mechanisms responsible for PPCPs removal during sand filtration.


Use of Retroauricular Flaps to Reconstruct Defects of the Helical Rim and Adjuvant Intralesional Corticosteroids after Resection of Large Keloids.
The optimal treatment for large helical rim keloids remains unclear. The authors evaluated patient outcomes when retro auricular flaps were used to reconstruct helical rim defects and adjuvant intralesional corticosteroids were prescribed after resection of large keloids. The authors evaluated 7 females with 12 large keloids of the helical rim (5 patients had lesions in both ears). All patients were aged 18 to 33 years. The authors used retro auricular flaps to reconstruct the helical rim defects and prescribed adjuvant intralesional corticosteroids after resection. The lesion area ranged from 2.5 × 2.0 to 3.5 × 3.5 cm2. The flap size ranged from 2.5 × 3.0 to 3.5 × 4.5 cm2. The flaps and wound bases received 3 steroid injections at approximately 1, 2, and 3 months postoperatively. No flap necrosis or complications were noted. The postoperative esthetic results were satisfactory in 8 patients and excellent in 4. All patients were followed up for 14 to 28 months (median, 20.6 months). No recurrence was noted, although 3 patients exhibited mild scarring of the wound flap base. A retro auricular flap is safe and effective for reconstructing helical rim defects; adjuvant intralesional corticosteroids prevent scarring of the flap wound base after resection of large keloids.


Minimal preconditioning effects observed for inflation tests of planar tissues.
The purpose of this study is to investigate the effects of preconditioning on the deformation response of planar tissues measured by inflation tests. The inflation response of test specimens, including the bovine cornea, bovine and porcine sclera, and human skin, exhibited a negligible evolving deformation response when subjected to repeated pressure loading with recovery periods between cycles. Tissues obtained complete recovery to the reference state, and strain contours across the entire specimen were nearly identical at the maximum pressure of each load cycle. This repeatability was obtained regardless of strain history. These results suggest that negligible permanent change was induced in the microstructure by inflation testing. Additionally, we present data illustrating that a lack of a recovery period can result in an evolving deformation response to repeated loading that is commonly attributed to preconditioning. These results suggest that the commonly observed effects of preconditioning may be avoided by experimental design for planar tissues characterized by long collagen fibers arranged in the plane of the tissue. Specifically, if the test is designed to fully fix the specimen boundary during loading, adequate recovery periods are allowed after each load cycle, and loads are limited to avoid damage, preconditioning effects may be avoided for planar tissues.


Microsporidium in AIDS patients: a perspective.
Microsporidia are ubiquitous, obligate intracellular protozoan parasites increasingly detected as opportunistic pathogens in AIDS patients. These parasites have been associated with chronic diarrhoea, hepatitis, cholangitis, pancreatitis, enteritis, keratoconjunctivitis, and peritonitis in either homosexuals or heterosexuals. Optimum diagnostic and therapeutic measures of these pathogens still elude both clinicians and researchers. Further study is required to elucidate the exact prevalence and clinical characteristics of microsporidia.


Effect of medicare payment on rural health care systems.
Medicare payments constitute a significant share of patient-generated revenues for rural providers, more so than for urban providers. Therefore, Medicare payment policies influence the behavior of rural providers and determine their financial viability. Health services researchers need to contribute to the understanding of the implications of changes in fee-for-service payment policy, prospects for change because of the payment to Medicare+Choice risk plans, and implications for rural providers inherent in any restructuring of the Medicare program. This article outlines the basic policy choices, implications for rural providers and Medicare beneficiaries, impacts of existing research, and suggestions for further research. Topics for further research include implications of the Critical Access Hospital program, understanding how changes in payment to rural hospitals affect patient care, developing improved formulas for paying rural hospitals, determining the payment-to-cost ratio for physicians, measuring the impact of changes in the payment methodology used to pay for services delivered by rural health clinics and federally qualified health centers, accounting for the reasons for differences in historical Medicare expenditures across rural counties and between rural and urban counties, explicating all reasons for Medicare+Choice plans withdrawing from some rural areas and entering others, measuring the rural impact of proposals to add a prescription drug benefit to the Medicare program, and measuring the impact of Medicare payment policies on rural economies.


Molecular and functional analyses of the metC gene of Lactococcus lactis, encoding cystathionine beta-lyase.
The enzymatic degradation of amino acids in cheese is believed to generate aroma compounds and therefore to be essential for flavor development. Cystathionine beta-lyase (CBL) can convert cystathionine to homocysteine but is also able to catalyze an alpha, gamma elimination. With methionine as a substrate, it produces volatile sulfur compounds which are important for flavor formation in Gouda cheese. The metC gene, which encodes CBL, was cloned from the Lactococcus lactis model strain MG1363 and from strain B78, isolated from a cheese starter culture and known to have a high capacity to produce volatile compounds. The metC gene was found to be cotranscribed with a downstream cysK gene, which encodes a putative cysteine synthase. The MetC proteins of both strains were overproduced in strain MG1363 with the NICE (nisin-controlled expression) system, resulting in a >25-fold increase in cystathionine lyase activity. A disruption of the metC gene was achieved in strain MG1363. Determination of enzymatic activities in the overproducing and knockout strains revealed that MetC is essential for the degradation of cystathionine but that at least one lyase other than CBL contributes to methionine degradation via alpha, gamma elimination to form volatile aroma compounds.


Stakeholder perceptions of bird-window collisions.
Bird-window collisions are a major source of human-caused avian mortality for which many mitigation and prevention options are available. However, because very little research has characterized human perspectives related to this issue, there is limited understanding about the most effective ways to engage the public in collision reduction efforts. To address this research need, we: (1) evaluated how two stakeholder groups, homeowners and conservation practitioners, prioritize potential benefits and obstacles related to bird-window collision management, (2) compared priorities between these groups, and (3) evaluated potential conflicts and collective strength of opinions within groups. We addressed these objectives by merging the strengths, weaknesses, opportunities, and threats (SWOT) and analytic hierarchy process (AHP) survey approaches. Specifically, survey respondents made pairwise comparisons between strengths and weaknesses (respectively, direct outcomes and barriers related to management, such as fewer collisions and increased costs) and opportunities and threats (indirect outcomes and barriers, such as increased bird populations and fewer resources for other building-related expenses). Both homeowners and conservation practitioners ranked strengths and opportunities higher than weaknesses and threats, indicating they have an overall positive perception toward reducing bird-window collisions. However, key obstacles that were identified included costs of management and a lack of policy and guidelines to require or guide management. These results suggest that substantial advances can be made to reduce bird-window collisions because both homeowners and conservation practitioners had positive views, suggesting their receptivity toward collision management measures. However, because of more neutral views and conflicting responses within the homeowner group, results also highlight the importance of targeting homeowners with education materials that provide information about bird-window collisions and solutions that reduce them. Because bird-window collisions are a human-caused phenomenon, such information about human perspectives and priorities will be crucial to addressing this threat and thus benefitting bird populations.


Overview: "Children's toxicology", a renovating study field of irreversible "early exposure-delayed effects".
"Children are not small adults". This is a well-known phrase, especially in the clinics for diagnosis, efficacy of treatment, side effect, and prognosis. However, in the field of toxicology, this issue has long been a challenge. The knowledge has been limited to the differences in metabolism and other physiological factors. Currently available test guidelines for fetuses and immature animals are teratogenicity and reproductive toxicity studies. These tests look for straight-forward (essentially macroscopic) outcomes established within a rather short period of exposure to the test substances. However, recent advances in molecular toxicology allow combination of in vitro and in vivo studies at molecular levels. The target molecules and receptors can be identified in quantitative fashion and at the fine structure levels around and below the resolution of normal light microscopy. Such expansion of the knowledge lead us to consider a rather new category of "receptor mediated toxicity" or "signal toxicity". Such non-organic insults would merely induce transient effects on adults. However, there are growing evidences that such slight insults on the developing and maturating organisms can leave irreversible effects that become overt in adulthood. As an overview, toxicology has entered a new phase where children's toxicology becomes a renovating study field of the irreversible "early exposure-delayed effects".


Rat renal function four days after bile-duct ligation: effects of indomethacin and vasoactive agents.
The purpose of this study was to assess the effects of the cyclooxygenase inhibitor, indomethacin, and some vasoactive agents on the renal functional parameters during the early liver injury induced by four days bile duct ligation (BDL). Wistar rats with four days-BDL and control-sham operated were used. Renal function was measured in anesthetized rat treated with a single dose of indomethacin (control, 0.3, 1.0, 3.0 mg/kg b.w.; i.v.) one hour before clearance studies. Sulindac effects were also evaluated (5 mg/kg b.w., i.p). Isolated rat kidney preparations from control and BDL donor rats were used to study renal vascular response to noradrenaline, carbachol or sodium nitroprusside. The bile duct ligation promoted a diminished renal cortical plasma flow (RCPF) on the fourth day post surgery accompanied with a diminution in the glomerular filtration rate (GFR), increased filtration fraction and increased fractional excretion of water and sodium. Indomethacin 0.3 mg/kg induced an increase in GFR and RCPF, maintaining the high filtration fraction in BDL rats. The other doses did not alter these parameters as compared with bile duct ligated rats without treatment, but indomethacin 3 mg/kg caused a significant increase in filtration fraction. Indomethacin induced dose-dependent diminution in natriuresis in sham and BDL groups. Sulindac did not modify hemodynamic parameters, but induced antinatriuresis and antidiuresis in both experimental groups. Maximal vascular responses to noradrenaline measured in isolated rat kidneys were statistically diminished in BDL-rats as compared with controls (C, n=7; 35.0+/-2.3 mmHg ml(-1) min; BDL-rats, n=5; 23.8+/-0.7 mmHg ml(-1) min; p<0.02), without changes in EC15. Maximal relaxation induced by sodium nitroprusside in the phenylephrine (PHE)-pre-constricted renal vasculature in control preparations did not differ from that observed in BDL group (C: n=6; 49.5+/-2.3%). Values of EC50 were 1.26+/-0.07 microM (n=6) in control preparations and 0.34+/-0.03 microM (n=4) in kidneys from BDL-rats (p<0.001). Carbachol induced a biphasic relaxation of PHE-pre-constricted renal vasculature. No differences in maximal responses were found. EC50 value of the second phase in BDL group was significantly decreased compared to control preparations (C: n=6, 0.47+/-0.05 microM; BDL: n=6, 0.22+/-0.03 microM p<0.001). The present results show that the altered renal function after a short time post bile duct ligation is determined, at least in part, by increased release of arachidonic derivatives in vascular bed and tubular cells. At this stage of liver injury, the alteration in the renal vascular response to different vasoactive agents is remarkable.


Isolation and Partial Characterization of a Factor from Barley Aleurone that Modifies alpha-Amylase in Vitro.
Posttranslational modifications that give rise to multiple forms of alpha-amylase (EC 22.214.171.124) in barley (Hordeum vulgare L. cv Himalaya) were studied. When analyzed by denaturing polyacrylamide gel electrophoresis, barley alpha-amylase has a molecular mass of 43 to 44 kilodaltons, but isoelectric focusing resolves the enzyme into a large number of isoforms. To precisely identify these isoforms, we propose a system of classification based on their isoelectric points (pl). alpha-Amylases with pls of approximately 5, previously referred to as low pl or Amy1 isoforms, have been designated HAMY1, and alpha-amylases with pls of approximately 6, referred to as high pl or Amy2, are designated HAMY2. Individual isoforms of HAMY1 and HAMY2 are identified by their pls. For example, the most acidic alpha-amylase synthesized and secreted by barley aleurone layers is designated HAMY1(4.56). Some of the diversity in the pls of barley alpha-amylases arises from posttranslational modifications of the enzyme. We report the isolation of a factor from barley aleurone layers and incubation media that can modify HAMY1 isoforms in vitro. This factor has a molecular mass between 30 and 50 kilodaltons, and it can catalyze the conversion of HAMY1(4.90) and HAMY1(4.64) to isoforms 4.72 and 4.56, respectively. The in vitro conversion of HAMY1 isoforms by the factor is favored by pH values of approximately 5 and is inhibited at approximately pH 7. The level of this factor in aleurone layers and incubation media is not affected by treatment of the tissue with gibberellic acid. The amylase-modifying activity from barley will also modify alpha-amylases isolated from human saliva and porcine pancreas. An activity that can modify HAMY1 isoforms in vitro has also been isolated from Onozuka R10 cellulase. Because the activity isolated from barley lowers the pl of alpha-amylase from barley, human saliva, and porcine pancreas, we speculate that it is a deamidase.


Five fully automated methods for performing immature reticulocyte fraction: comparison in diagnosis of bone marrow aplasia.
We performed a parallel evaluation of 5 automated reticulocyte counters to produce the immature reticulocyte fraction (IRF). We analyzed 225 samples from healthy control subjects, 115 from patients with various diseases, 38 with advanced aplasia, and 22 in early erythropoietic recovery after chemotherapy or bone marrow transplantation. The reference intervals were different for each instrument (ADVIA 120, 0.04-0.25; CELL DYN 4000, 0.15-0.35; GEN-S, 0.20-0.37; SE 9500 RET 0.05-0.21; VEGA RETIC: 0.06-0.23). The imprecision, obtained by 1-way analysis of variance on duplicates, was satisfactory for clinical use for all methods (coefficient of variation, 7.6%-20.5% in healthy subjects), although it was higher than the analytic goal based on biologic variability within subjects. The comparison of different methods shows that agreement is good only between SE 9500 RET CELL DYN 4000, and VEGA RETIC (r2 = 0.72-0.78). The study of diagnostic performance in distinguishing aplasia from early bone marrow recovery shows slightly different results (area under the curve from 0.70 for ADVIA 120 to 0.96 for SE 9500 RET). Even with slight differences, the fluorescence-based methods seem to be more robust than other methods for IRF measurement.


Trial-to-trial modulations of the Simon effect in conditions of attentional limitations: Evidence from dual tasks.
Interference effects are reduced after trials including response conflict. This sequential modulation has often been attributed to a top-down mediated adaptive control mechanism and/or to feature repetition mechanisms. In the present study we tested whether mechanisms responsible for such sequential modulations are subject to attentional limitations under dual-task situations. Participants performed a Simon task in mixed single- and dual-task contexts (Experiment 1), in blocked contexts with dual-task load either, in trialN (Experiment 2a), in trialN-1 (Experiment 2b), or in both trials (Experiment 3). Results showed that the occurrence of a sequential modulation did not depend on dual-task load per se as it occurred predominantly in conditions of lowest and highest task load. Instead, task factors such as the repetition of task episodes and stimulus-response repetitions determined whether a sequential modulation occurred.


The small GTPase Rab13 regulates assembly of functional tight junctions in epithelial cells.
Junctional complexes such as tight junctions (TJ) and adherens junctions are required for maintaining cell surface asymmetry and polarized transport in epithelial cells. We have shown that Rab13 is recruited to junctional complexes from a cytosolic pool after cell-cell contact formation. In this study, we investigate the role of Rab13 in modulating TJ structure and functions in epithelial MDCK cells. We generate stable MDCK cell lines expressing inactive (T22N mutant) and constitutively active (Q67L mutant) Rab13 as GFP-Rab13 chimeras. Expression of GFP-Rab13Q67L delayed the formation of electrically tight epithelial monolayers as monitored by transepithelial electrical resistance (TER) and induced the leakage of small nonionic tracers from the apical domain. It also disrupted the TJ fence diffusion barrier. Freeze-fracture EM analysis revealed that tight junctional structures did not form a continuous belt but rather a discontinuous series of stranded clusters. Immunofluorescence studies showed that the expression of Rab13Q67L delayed the localization of the TJ transmembrane protein, claudin1, at the cell surface. In contrast, the inactive Rab13T22N mutant did not disrupt TJ functions, TJ strand architecture nor claudin1 localization. Our data revealed that Rab13 plays an important role in regulating both the structure and function of tight junctions.


Characterisation of dog allergens by means of immunoblotting.
Sera from 75 patients with clinical type I allergy against dogs were investigated by means of immunoblotting using extracts prepared from dog hair/dander (CAN XI D) and saliva. In addition, selected sera were tested on extracts made of hair, skin, salivary glands (parotis and submandibularis), serum and liver. A 69-kD IgE-binding protein was identified in all extracts tested with an incidence of approximately 40% and shown to be dog albumin by means of inhibition experiments. In 96% of patients' sera IgE antibodies reactive with a 19-kD and/or a 23-kD protein of the hair/dander extract (CAN XI D) were observed. IgE binding to a 23-kD band was also detected in the hair and saliva extracts, but not in skin, salivary gland, serum and liver extracts. A 19-kD IgE-binding protein was strongly expressed in skin and to a lesser degree in saliva, but not in hair, serum and liver. Preincubation of patients sera with the hair extract and subsequent probing with the hair/dander extract (CAN XI D) inhibited IgE binding to the 23 kD protein whereas preincubation with the skin extract abolished IgE binding to the 19-kD protein. Using the hair/dander extract as inhibitor, IgE binding to the 19- and 23-kD proteins of saliva was abrogated. Thus it is concluded that the 23-kD protein is preferentially expressed in hair and saliva whereas the 19-kD protein is found in saliva and skin. Furthermore these two proteins are likely to represent immunologically independent major allergens.


Use of the stationary phase inducible promoters, spv and dps, to drive heterologous antigen expression in Salmonella vaccine strains.
We have investigated the ability of the growth phase regulated promoters dps and spv, to drive expression of heterologous antigens in Salmonella vaccine strains. Reporter plasmids were constructed which directed beta-galactosidase expression from dps (pDpslacZ) or spv (pSpvlacZ) and these were introduced independently into the Salmonella typhimurium vaccine strain SL3261 (aroA(-)). beta-galactosidase expression was induced 20-fold and 100-fold when broth cultures of SL3261 (pDpslacZ) or SL3261 (pSpvlacZ) respectively, entered the stationary phase of growth. Within macrophages, beta-galactosidase expression was induced 3.5-fold with SL3261 (pDpslacZ) and 7-fold with SL3261 (pSpvlacZ). The spv and dps promoters were used to drive independent expression of the C fragment domain of tetanus toxin (TetC) from plasmids harboured in S. typhimurium SL3261. Levels of anti-TetC antibodies were significantly higher in the sera of BALB/c mice perorally inoculated with SL3261 (pSpvtetC) or SL3261 (pDpstetC) compared to unvaccinated controls. This suggests that these promoter systems may be used to drive foreign antigen expression in live oral Salmonella vaccines.


Evaluation of Neurocognitive Abilities in Patients Undergoing Carotid Endarterectomy Surgery.
To evaluate the differences in neurocognitive abilities between the preoperative and postoperative periods following carotid endarterectomy (CEA), due to carotid artery stenosis, and to evaluate the effectiveness of CEA on neurocognitive abilities in the future. Thirty-eight cases of CEA surgery at Bozok University Faculty of Medicine Research Hospital between January 2015 and June 2020 were examined. Neurocognitive tests were performed on carotid endarterectomy patients one day before the operation and on the 2nd, 4th, and 30th postoperative days. The effect of CEA on cognitive results has been investigated. Of the patients, eight were female (21.1%), 30 were male (78.9%), and the mean age was 66 ± 4.09. Thirty-two (84.21%) of the patients were operated on under general anesthesia and six (15.78%) under regional anesthesia. A shunt was used in 19 patients. Right carotid endarterectomy was performed in 20 cases and left carotid endarterectomy in 18 cases. We used the primary closure technique in two of 38 cases and patches on 36 of them. We used Dacron in 21 cases, PTFE in 12 cases, and saphenous vein as a patch in three cases. In the WMS digit spam and recall scores, the postoperative period fell on the 2nd day, and then on the 4th and 30th day after the operation, there was a low level of increase over time. Compared with the preoperative period, the learning score was found to be the lowest on the 2nd day, lower on the 4th day compared with the preoperative period and improved compared with the preoperative period on the 30th day. There was no decrease in the verbal fluency test score results after the operation, on the contrary, it was observed minimally. The test score results cumulatively were decreased in the early postoperative periods compared with the preoperative period and increased on the 30th day compared with the preoperative period. The purpose of CEA in the past was the prevention of ischemic stroke and cerebrovascular disease (CVD) rather than neurocognitive recovery. Factors affecting neurocognition in CEA are multifactorial. Preservation and improvement of neurocognition are more important than any other period of history. By prioritizing cognitive abilities in the treatment of carotid stenosis, individualization of the treatment will help maximize the increase in cognitive abilities by providing optimum benefit to the patient of each factor.


Photocatalytic Activity and Fluorescence of Gold/Zinc Oxide Nanoparticles Formed by Dithiol Linking.
Monolayer-protected gold nanoparticles (AuNPs) with average diameters of 2-4 nm have been covalently attached to zinc oxide nanorods using dithiol ligands. Electron microscopy and Raman spectroscopy show that ozone treatment or annealing at 300 or 450 °C increases the average diameter of the AuNPs to 6, 8, and 14 (±1) nm, respectively, and decomposes the organic layers to various degrees. These treatments locate the AuNPs closer to the nanorods. Heating and subsequent ozone exposure changes the color of the as-prepared nanocomposite powder from blue to purple due to oxidation of the outer layer of the AuNPs, and heating to 300 °C changes it to pink due to oxygen desorption. ZnO nanorods have a bimodal photoluminescence spectrum that consists of an ultraviolet excitonic peak and a visible, surface defect-related peak. Ozone treatment and annealing of the nanocomposite decreases the intensities of both peaks due to quenching by the AuNPs, but the visible peak is affected less. The photocatalytic efficiency of the nanocomposites toward oxidative degradation of rhodamine B has been measured and follows the order 300 °C > 450 °C > ozone treated ≈ as-prepared ≈ bare ZnO. The greater efficiency of the annealed samples likely arises from decreased electron-hole pair recombination rates.


Cytochrome c-peroxidase modulates ROS homeostasis to regulate the sexual mating of Sporisorium scitamineum.
Reactive oxygen species play an important role in pathogen-plant interactions. In fungi, cytochrome c-peroxidase maintains intracellular ROS homeostasis by utilizing H2O2 as an electron acceptor to oxidize ferrocytochrome c, thereby contributing to disease pathogenesis. In this study, our investigation reveals that the cytochrome c-peroxidase encoding gene, SsCCP1, not only plays a key role in resisting H2O2 toxicity but is also essential for the mating/filamentation and pathogenicity of S. scitamineum. We further uncover that SsCcp1 mediates the expression of SsPrf1 by maintaining intracellular ROS homeostasis to regulate S. scitamineum mating/filamentation. Our findings provide novel insights into how cytochrome c-peroxidase regulates sexual reproduction in phytopathogenic fungi, presenting a theoretical foundation for designing new disease control strategies.


Urinary trypsin inhibitor concentration can predict the immunological insult of chemotherapy and complications after bone marrow transplantation.
Urinary trypsin inhibitor has attracted attention as an index of the systemic inflammatory response syndrome. In this study, the urine concentration of trypsin inhibitor was measured to compare the immunological insult of conventional chemotherapy and conditioning chemotherapy for bone marrow transplantation. We also investigated whether urinary trypsin inhibitor was a useful index of the complications and outcome of bone marrow transplantation. Urinary trypsin inhibitor concentration was determined before chemotherapy, on the day after finishing chemotherapy (day 0 of transplantation), and during recovery of the white cell count, in 17 patients (seven receiving conventional chemotherapy and 10 receiving conditioning for bone marrow transplantation). Urinary trypsin inhibitor concentrations were significantly higher after conditioning for bone marrow transplantation than after conventional chemotherapy (P < 0.001), indicating that conditioning was more invasive. After bone marrow transplantation, the incidence of severe complications and the mortality rate were higher in patients whose urinary trypsin inhibitor concentrations rose during recovery of the white cell count. Comparison of urinary trypsin inhibitor concentrations suggested that conditioning for bone marrow transplantation was more invasive than conventional chemotherapy. This study also suggested that the urine concentration of trypsin inhibitor could be useful for predicting the risk of complications and outcome of bone marrow transplantation.


Exploring the Barriers to Breast and Cervical Cancer Screening in Nigeria: A Narrative Review.
Breast and cervical cancers are the most common causes of female cancer-related mortality in Nigeria. Early detection and treatment significantly decreases cancer mortality rates. Various factors influence uptake of cancer screening. Cancer awareness, availability of screening, and treatment facilities alone have not been completely successful in improving cancer health behaviour. This review aims to identify and summarise the barriers to breast and cervical cancer screening uptake in Nigeria. Various databases such as PubMed, Psych Info, Google Scholar and EMBASE were extensively searched for existing literature on factors influencing breast and cervical cancer screening in Nigeria. Studies retrieved explored the major socioeconomic factors affecting women's knowledge, perception, and attitude. Embarrassment, low perception of cancer risk, and physician gender preference are some of the most common factors that discouraged women from cancer screening. Lack of spouse permission and support; belief that cancer is a death wish, and societal discrimination are the common sociocultural barriers to screening. These factors vary across different regions in Nigeria. Therefore, policy makers should make deliberate effort to develop cancer management strategies that are tailored to the sociocultural and religious needs in a target population. This approach is anticipated to improve uptake and ensure sustainability of cancer management.


Serum interleukin-6 and endotoxin levels and their relationship with fatigue and depressive symptoms in patients on chronic haemodialysis.
Little is known about the underlying aetiology of fatigue in haemodialysis (HD) patients apart from a significant association and overlapping symptomatology with depressive symptoms. Growing evidence exists for the involvement of the immune system, by pro-inflammatory cytokines, in the development of fatigue in other inflammatory diseases. In HD patients, increased exposure to bacterial endotoxins may contribute to an inflammatory response and may potentially lead to fatigue. We therefore aimed (i) to assess the interrelationship between serum endotoxin (EA) levels, interleukin-6 (IL-6) levels and fatigue in HD patients; (ii) to evaluate whether there is a relationship between depressive symptoms and inflammation as well and (iii) to what extent depressive symptoms and fatigue are related to each other. Fatigue and depressive symptoms in daily life were assessed in 59 individuals using the SF-36 vitality subscale and the Geriatric Depression Scale. Blood samples were collected on a mid-week dialysis session to determine EA levels, through the Endotoxin Activity Assay (EAA™), and IL-6 concentrations, through the commercially available Abcam ELISA (Enzyme-Linked Immunosorbent Assay) kit. EA, IL-6 levels and depressive symptoms were significantly correlated with fatigue. EA levels and depressive symptoms were significant predictors of fatigue, explaining 31% of its variance. However, EA and IL-6 were not significantly associated with depression. Fatigue in HD patients may be related to endotoxemia and inflammation through IL-6. Furthermore, fatigue is significantly associated with depressive symptoms. Future research into the causal interrelationship of inflammation, fatigue and depression in HD patients might lead to potential targets for therapeutic strategies.


[Degradation of cyanobacterial peptide hepatotoxins by ferrate].
In this study, the effect of ferrate oxidation on stability of microcystins-LR (MCLR) in freeze-dried Oscillatoria amoena was investigated. The toxin was easily decomposed by oxidation with ferrate, and the stability depended on the dosage of ferrate and pH. Simultaneously the product Fe(OH)3 after ferrate oxidation could flocculate the organic compounds so that 50% TOC removal of the extract was reached. The low residual iron reflected the excellent results of coagulation and could meet the standard of drinking water. The results show that ferrate oxidation may be an effective and practical method for the removal of cyanobacterial peptide toxins from drinking water.


NMDA receptor subunits epsilon 1 (NR2A) and epsilon 2 (NR2B) are substrates for Fyn in the postsynaptic density fraction isolated from the rat brain.
Fyn, protein tyrosine kinase, and its substrates were highly concentrated in the postsynaptic density (PSD) fraction prepared from the rat forebrain. There were a number of Fyn substrates unique to the PSD fraction. One of the major substrates in the PSD fraction was found to be a concanavalin A-binding glycoprotein, PSD-gp180, which is the N-methyl-D-aspartate (NMDA) receptor subunit epsilon 2 (NR2B). Western blotting and immunoprecipitation supported the phosphorylation of epsilon 2 by Fyn. NMDA receptor subunit epsilon 1 (NR2A) was also a substrate for Fyn. These results suggest that Fyn is involved in the modulation of synaptic efficacy through the phosphorylation of synapse-specific substrates such as the NMDA receptor/channel.


Re-dissolution and de-compaction of DNA-cationic surfactant complexes using non-ionic surfactants.
Addition of a cationic surfactant to a solution of DNA causes the formation of compacted DNA-cationic surfactant complexes which precipitate from aqueous solution. It has been shown previously that addition of anionic surfactant will re-dissolve and de-compact the DNA-cationic surfactant complexes and we find that addition of non-ionic surfactants of the alkylpolyoxyethylene type can be used similarly. In principle, these de-compaction and re-dissolution processes could occur either by stripping of the cationic surfactant from the DNA into mixed micelles with the non-ionic surfactant or by solubilisation of the DNA-cationic surfactant complexes within the non-ionic micelles. Solubility phase-boundary measurements, fluorescence microscopy observations of the de-compaction process and light scattering results indicate that de-compaction and re-dissolution occur by the stripping mechanism, even for non-ionic surfactants where the favourable attractive electrostatic interaction between the two surfactants is absent. Using measurements of critical micelle concentrations and calculations based on regular solution mixed micelle theory, we show that re-dissolution and de-compaction of the DNA-cationic surfactant complexes occurs when the concentration of free monomeric cationic surfactant is reduced (by incorporation into mixed micelles) below a critical value.


Novel Treatment Approach for Deep Palmoplantar Warts Using Long-Pulsed 1064-nm Nd:YAG Laser and a Moisturizing Cream Without Prior Paring of the Wart Surface.
The present study aimed to assess the safety and efficacy of palmoplantar wart removal using long-pulsed 1064-nm Nd:YAG laser after application of a moisturizing cream. Previously described laser treatments for wart removal are associated with negative side effects and need to pare the warts before laser treatment. Two hundred forty patients (142 males, 98 females) were treated for 1-40 palmoplantar warts by long-pulsed 1064-nm Nd:YAG laser (spot size 4-6 mm, pulse duration 20 msec, fluence 200 J/cm2) after covering the wart surface with a thin film of a moisturizing cream. The endpoint was lesion graying or whitening with or without development of a hemorrhagic bulla beneath the treated wart. Color photographs were taken before and immediately after each laser session and at 1, 4, and 16 weeks after the last session. The overall clearance rate was 97%, with 90% of treated patients cured after one session, 4% after two, and 3% after three. Clearance rate after three laser sessions decreased linearly with the number of warts from 100% to 95%. Less accessible wart location in interdigital spaces also decreased the cure rate after three sessions from 100% to 95%. Additionally, warts became more difficult to eradicate as they aged. Remission lasted up to 6 years, and complications were mild and infrequent (17.5%). This novel method is effective in removing palmoplantar warts. It is easier, time-saving, and safer than other methods described in previous studies conducted with ablative or nonablative lasers.


Myocardial strain characteristics and outcomes after transcatheter aortic valve replacement.
Objective of this study was to make an assessment of standard functional and defor-mation parameters (strain) in patients after transcatheter aortic valve replacement (TAVR) by cardiac magnetic resonance imaging (CMR) and the evaluation of their prognostic impact. Patients undergoing TAVR received CMR on a 1.5 T whole-body scanner at 3 months after the procedure. Deformation parameters (strain, strain rate, velocity, displacement) were assessed in lon-gitudinal, circumferential and radial orientation using a feature tracking approach. Primary outcome measure was defined according to Valve Academic Research Consortium-2 (VARC-2) criteria. Eighty-three patients formed the study population. Deformation parameters were significantly reduced in all three orientations for strain (longitudinal: -12.1 ± 5.4% vs. -15.9 ± 1.96%, p < 0.0001; radial: 34.4 ± 15.3% vs. 47.2 ± 11.4%, p < 0.0001; circumferential: -16.8 ± 4.3% vs. -21.1 ± 2.5%, p < 0.0001) and strain rate (longitudinal: -0.79 ± 0.33%/s vs. -0.91 ± 0.23%/s, p = 0.043; radial: 2.5 ± 1.2%/s vs. 2.9 ± 0.9%, p = 0.067; circumferential: -1.1 ± 0.6%/s vs. -1.3 ± 0.3%/s, p = 0.006) in comparison to a healthy control population. Median follow-up was 614 days. During this period, 13 endpoints occurred (cumulative event rate of 10.7%). Patients with event by trend exhibited poorer strain and strain rate in longitudinal and radial orientation without reaching statistical significance (longitudinal strain: -11.2 ± 5.4% vs. -12.3 ± 5.4%, p = 0.52; longitudinal strain rate: -0.73 ± ± 0.23%/s vs. 0.80 ± 0.35%/s, p = 0.53; radial strain: 29.5 ± 19.6% vs. 35.2 ± 14.5%, p = 0.24; radial strain rate: 2.2 ± 1.6%/s vs. 2.6 ± 1.2%/s, p = 0.31). Assessment of left ventricular deformation parameters by CMR revealed functional abnormalities in comparison to healthy controls. Prognostic significance remains to be further investi-gated.


Constructing Precisely Computing Networks with Biophysical Spiking Neurons.
While spike timing has been shown to carry detailed stimulus information at the sensory periphery, its possible role in network computation is less clear. Most models of computation by neural networks are based on population firing rates. In equivalent spiking implementations, firing is assumed to be random such that averaging across populations of neurons recovers the rate-based approach. Recently, however, Denéve and colleagues have suggested that the spiking behavior of neurons may be fundamental to how neuronal networks compute, with precise spike timing determined by each neuron's contribution to producing the desired output (Boerlin and Denéve, 2011; Boerlin et al., 2013). By postulating that each neuron fires to reduce the error in the network's output, it was demonstrated that linear computations can be performed by networks of integrate-and-fire neurons that communicate through instantaneous synapses. This left open, however, the possibility that realistic networks, with conductance-based neurons with subthreshold nonlinearity and the slower timescales of biophysical synapses, may not fit into this framework. Here, we show how the spike-based approach can be extended to biophysically plausible networks. We then show that our network reproduces a number of key features of cortical networks including irregular and Poisson-like spike times and a tight balance between excitation and inhibition. Lastly, we discuss how the behavior of our model scales with network size or with the number of neurons "recorded" from a larger computing network. These results significantly increase the biological plausibility of the spike-based approach to network computation. We derive a network of neurons with standard spike-generating currents and synapses with realistic timescales that computes based upon the principle that the precise timing of each spike is important for the computation. We then show that our network reproduces a number of key features of cortical networks including irregular, Poisson-like spike times, and a tight balance between excitation and inhibition. These results significantly increase the biological plausibility of the spike-based approach to network computation, and uncover how several components of biological networks may work together to efficiently carry out computation.


Constructing firefly luciferin bioluminescence probes for in vivo imaging.
Bioluminescence imaging (BLI) is a widely applied visual approach for real-time detecting many physiological and pathological processes in a variety of biological systems. Based on the caging strategy, lots of bioluminescent probes have been well developed. While the targets react with recognizable groups, caged luciferins liberate luciferase substrates, which react with luciferase generating a bioluminescent response. Among the various bioluminescent systems, the most widely utilized bioluminescent system is the firefly luciferin system. The H and carboxylic acid of luciferin are critically caged sites. The introduced self-immolative linker extends the applications of probes. Firefly luciferin system probes have been successfully applied for analyzing physiological processes, monitoring the environment, diagnosing diseases, screening candidate drugs, and evaluating the therapeutic effect. Here, we systematically review the general design strategies of firefly luciferin bioluminescence probes and their applications. Bioluminescence probes provide a new approach for facilitating investigation in a diverse range of fields. It inspires us to explore more robust light emission luciferin and novel design strategies to develop bioluminescent probes.


Alloxazine-cyclodextrin conjugates for organocatalytic enantioselective sulfoxidations.
Four structurally different alloxazine-cyclodextrin conjugates were prepared and tested as catalysts for the enantioselective oxidation of prochiral sulfides to sulfoxides by hydrogen peroxide in aqueous solutions. The alloxazinium unit was appended to the primary face of α- and β-cyclodextrins via a linker with variable length. A series of sulfides was used as substrates: n-alkyl methyl sulfides (n-alkyl = hexyl, octyl, decyl, dodecyl), cyclohexyl methyl sulfide, tert-butyl methyl sulfide, benzyl methyl sulfide and thioanisol. α-Cyclodextrin conjugate having alloxazinium unit attached via a short linker proved to be a suitable catalyst for oxidations of n-alkyl methyl sulfides, displaying conversions up to 98% and enantioselectivities up to 77% ee. β-Cyclodextrin conjugates were optimal catalysts for the oxidation of sulfides carrying bulkier substituents; e.g. tert-butyl methyl sulfide was oxidized with quantitative conversion and 91% ee. Low loadings (0.3-5 mol%) of the catalysts were used. No overoxidation to sulfones was observed in this study.


Amplification of CFTR exon 9 sequences to multiple locations in the human genome.
Cloning and characterization of the cystic fibrosis transmembrane conductance regulator (CFTR) gene led to the identification and isolation of cDNA and genomic sequences that cross-hybridized to the first nucleotide binding fold of CFTR. DNA sequence analysis of these clones showed that the cross-hybridizing sequences corresponded to CFTR exon 9 and its flanking introns, juxtapositioned with two segments of LINE1 sequences. The CFTR sequence appeared to have been transcribed from the opposite direction of the gene, reversely transcribed, and co-integrated with the L1 sequences into a chromosome location distinct from that of the CFTR locus. Based on hybridization intensity and complexity of the restriction fragments, it was estimated that there were at least 10 copies of the "amplified" CFTR exon 9 sequences in the human genome. Furthermore, when DNA segments adjacent to the insertion site were used in genomic DNA blot hybridization analysis, multiple copies were also detected. The overall similarity between these CFTR exon 9-related sequences suggested that they were derived from a single retrotransposition event and subsequent sequence amplification. The amplification unit appeared to be greater than 30 kb. Physical mapping studies including in situ hybridization to human metaphase chromosomes showed that multiple copies of these amplified sequences (with and without the CFTR exon 9 insertion) were dispersed throughout the genome. These findings provide insight into the structure and evolution of the human genome.


Therapeutic strategies for the treatment of dyspepsia.
Dyspeptic symptoms are highly prevalent in the population and represent a major burden for healthcare systems. The ROME III criteria address and define two separate entities of functional dyspepsia: epigastric pain syndrome and postprandial distress syndrome. The etiology of dyspeptic symptoms is heterogeneous, underlying mechanisms are poorly understood and symptomatic improvement after drug therapy is often incomplete. This review of the literature included Medline data being published in the field of functional dyspepsia and different therapies. The reader will gain a current, unbiased understanding of the pathophysiological mechanisms underlying functional dyspepsia and of the therapeutic regimens based on randomized, controlled trials and on the meta-analyses that have been published on different therapeutic agents. Before starting medical treatment, a careful physical examination should exclude 'alarm symptoms'. Laboratory data, ultrasound and endoscopy are recommended in patients older than 45 - 55 years (depending on the guidelines being used). In areas with a high prevalence of Helicobacter pylori, the initial strategy includes 'test and treat' for H. pylori in addition to empiric acid suppressive therapy. Many studies have focused on the role of gastrointestinal dysmotility and hypersensitivity for dyspepsia with inconclusive results. Further therapeutic medical strategies include prokinetics, herbal preparations and psycho-/neurotopic drugs as well as additional psycho- or hypnotherapy.


Distribution and characterization of pedal peptide immunoreactivity in Aplysia.
Pedal peptide (Pep) is a very abundant neuropeptide in Aplysia. A radioimmunoassay (RIA) was developed to quantify Pep-like immunoreactivity (IR-Pep) in tissue extracts. IR-Pep was present in very high concentrations in the central nervous system (CNS) and two peripheral tissues: the large hermaphroditic duct (LHD) and the foot. RIA of fractions from high-pressure liquid chromatography (HPLC) indicated that Pep itself was the predominant immunoreactive species in each of these tissues. Lower concentrations of Pep were found in a number of other peripheral tissues. Incorporation of labelled amino acid indicated that Pep was synthesized in the LHD, whereas Pep in the foot was synthesized primarily in central neurons and transported to the foot. IR-Pep was further localized by immunocytology. All peripheral IR-Pep appeared to be associated with neuronal fibers, most commonly varicose axons. Immunoreactive innervation of the LHD and foot was particularly dense but positive staining was also observed in other tissues including tegument, gill, gut, and heart, IR-Pep innervation in all tissues including the LHD appeared to be localized predominantly in muscular portions of the tissue. Spontaneous contractions of isolated LHD were accelerated by the application of Pep. Pep appears to act as a transmitter or neuromodulator at a number of different sites in Aplysia.


Mono- and dimeric ferulic acid release from brewer's spent grain by fungal feruloyl esterases.
Ultraflo L, a beta-glucanase preparation from Humicola insolens sold for reducing viscosity problems in the brewing industry, exhibited activity against the methyl esters of ferulic, caffeic, p-coumaric and sinapic acids, displaying mainly type-B feruloyl esterase activity. Ultraflo also contained the ability to release 65% of the available ferulic acid (FA) together with three forms of diferulate from brewer's spent grain (BSG). An "esterase-free" Ultraflo preparation greatly enhanced the ability of a feruloyl esterase from Aspergillus niger, AnFAEA, to release FA (from 23 to 47%) and its dimeric forms, especially the 8,5' benzofuran form, from BSG. While total release of these phenolic acids was not observed, this synergistic enhancement of ferulate release demonstrates that FA and its dimeric forms present in BSG require the addition of more than a xylanase. This suggests either that FA is not solely attached to arabinoxylan in the barley cell wall, or that the cell wall polysaccharides in BSG hinder the accessibility of enzymes to the ferulates, due to processing treatments.


[Identification and expression of blaCTX-M-14 and blaCTX-M-24].
To identify the ESBL gene and the prevalence in Escherichia coli and Klebsiella pneumoniae strain isolated from Huashan Hospital, Shanghai. Isolates were confirmed as an ESBL producing strain by double-disk synergy test and NCCLS Confirmatory Test. Antibiotic susceptibilities were determined by standard agar dilution procedure on Mueller-Hinton agar. To determine whether the resistance was transferable, the conjugation experiment was performed; plasmids were isolated from clinical isolates and transcojugants. The partial bla(gene) of ESBL producing isolates and their transcojugants were detected by PCR using universal primers for TEM, SHV, CTX-M-1group, Toho-1group, CTX-M-13group respectively. The entire bla(CTX-M-13) group were amplified by PCR using the primers outside the Open Reading Frame (ORF) of CTX-M-13group beta-lactamases; the PCR products of entire bla(CTX-M-13)group were cloned into vector and the recombinant plasmids were transformed into the recipient strain for expression; the PCR products were also directly sequenced and analyzed; the clinical isolates of ESBL producers were detected by PFGE. ESBL producers were resistant to most beta-lactams and non-beta-lactams. Most transconjugants were obtained at frequency of 10(-4) approximately 10(-5) and resistance to non-beta-lactams was cotransferred with the ESBL activity to the transconjugant. A plasmid of about > 23.1 kb was obtained from each tansconjugant by plasmid extraction. Partial gene amplification products of CTX-M-13 group gene were obtained from isolates and their transconjugants. The bla(CTX-M-13)group from 4 transconjugants were identified as bla(CTX-M-14), and other six were bla(CTX-M-24); those ESBLs were mediated by plasmids (> 23.1 kb); the transformants producing CTX-M-14 or CTX-M-24 were resistant to most beta-lactams, which were much more resistant to cefotaxime than to ceftazidine; PFGE patterns of those isolates were different. clinical isolate of Escherichia coli and Klebsiella pneumoniae isolated from Huashan Hospital, Shanhai produced CTX-M-14 or CTX-M-24, which caused the isolate resistant to most beta-lactams; no clone spread in those isolates was found.


Perception of the cabin attitude changes in hypergravity.
The G-excess illusion is becoming increasingly recognized as a cause of aviation fatalities. Studies of this illusion have looked at perception of subjects' orientation by moving the head during hypergravity, but independent of the pilot's head movement with respect to aircraft. This illusion can also occur by aircraft motion, but this has not been studied extensively. The aim of this study was to investigate the subject's perception of orientation to the simulator cab attitude changes at 1.6 G without making any head movement with respect to the cab, and assess the feasibility of simulating the G-excess illusion on the ground with a centrifuge-like spatial disorientation simulator. The 1.6-G force field was provided by the gravitoinertial force (GIF) of the simulator when it made an off-center (planetary) rotation at a constant velocity of 130 degrees x s(-1). Eleven subjects' perceptions of orientation of the cab attitudes were collected respectively by their report before and after certain cab tilt, in a roll plane of 1.6 G. When the cab was tilted 20 degrees at 1.6 G, the subjects perceived the angle to be 48.6 +/- 39.4 degrees. Most subjects experienced an exaggerated sensation to the cab attitude changes in roll plane. G-excess illusion can be generated in a centrifuge-like device on the ground.


Identification and dating of the fountain pen ink entries on documents by ion-pairing high-performance liquid chromatography.
A novel approach for the identification and dating of the fountain pen ink entries on paper has been established by ion-pairing high-performance liquid chromatography (IP-HPLC). Twelve black and six red fountain inks have been collected, and their ink entries have been prepared by drawing lines on paper. The chromatographic conditions for separation of their dye components after extraction with solvents were optimized. Under the optimized conditions, the 18 fountain pen inks were differentiated individually by comparing the number of detectable main or minor dye components, and the relative peak intensities of each component. The ink entries were artificially and naturally aged, and the analysis results showed that the ink dye components were significantly decomposed when exposed to UV or fluorescent light compare to those of inks stored under natural condition. The changes of the relative peak height for the dye components were linearly related to the aging time, especially under natural aging conditions. The degradation characteristics of the dye components under different aging conditions provide scientific evidences for dating of the suspicious fountain pen ink entries on document.


Bioinspired synthesis of continuous titania coat with tunable nanofiber-based network structure on linear polyethylenimine-covered substrates.
We report using the substrates covered with self-organized linear polyethylenimine (PEI) layer as biomimetic template to direct the formation of high-quality titania coat with well-defined nanofiber-based network structure. The titania deposition was simply achieved by dipping the PEI substrates into aqueous solution of titanium bislactate under ambient conditions. We found that crystalline PEI layer on the substrates is important for achieving titania coat with nanofiber-based structure. Compared to the titania powder formed by solution deposition, the interface-mediated nanostructured titania exhibited dramatically improved thermostability with being able to maintain anatase phase in majority even at 900 degrees C. Moreover, the nanostructures of titania coat could be well controlled by simply adjusting the formation conditions of crystalline PEI layer on substrates. Because of high-efficiency photocatalysis of anatase titania, the nanostructured surface exhibited good photoresponsive surface wettability through hydrophobic modification and light irradiation.


Evidence for clinical, genetic and biochemical variability in spinal muscular atrophy with progressive myoclonic epilepsy.
Spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME) is a recently delineated, autosomal recessive condition caused by rare mutations in the N-acylsphingosine amidohydrolase 1 (acid ceramidase) ASAH1 gene. It is characterized by motor neuron disease followed by progressive myoclonic seizures and eventual death due to respiratory insufficiency. Here we report an adolescent female who presented with atonic and absence seizures and myoclonic jerks and was later diagnosed as having myoclonic-absence seizures. An extensive genetic and metabolic work-up was unable to arrive at a molecular diagnosis. Whole exome sequencing (WES) identified two rare, deleterious mutations in the ASAH1 gene: c.850G>T;p.Gly284X and c.456A>C;p.Lys152Asn. These mutations were confirmed by Sanger sequencing in the patient and her parents. Functional studies in cultured fibroblasts showed that acid ceramidase was reduced in both overall amount and enzymatic activity. Ceramide level was doubled in the patient's fibroblasts as compared to control cells. The results of the WES and the functional studies prompted an electromyography (EMG) study that showed evidence of motor neuron disease despite only mild proximal muscle weakness. These findings expand the phenotypic spectrum of SMA-PME caused by novel mutations in ASAH1 and highlight the clinical utility of WES for rare, intractable forms of epilepsy.