Effects of adolescent caffeine consumption on cocaine self-administration and reinstatement of cocaine seeking.
Caffeine consumption by children and adolescents has risen dramatically in recent years, yet the lasting effects of caffeine consumption during adolescence remain poorly understood. These experiments explore the effects of adolescent caffeine consumption on cocaine self-administration and seeking using a rodent model. Sprague-Dawley rats consumed caffeine for 28 days during the adolescent period. Following the caffeine consumption period, the caffeine solution was replaced with water for the remainder of the experiment. Age-matched control rats received water for the duration of the study. Behavioral testing in a cocaine self-administration procedure occurred during adulthood (postnatal days 62-82) to evaluate how adolescent caffeine exposure influenced the reinforcing properties of cocaine. Cocaine seeking was also tested during extinction training and reinstatement tests following cocaine self-administration. Adolescent caffeine consumption increased the acquisition of cocaine self-administration and increased performance on different schedules of reinforcement. Consumption of caffeine in adult rats did not produce similar enhancements in cocaine self-administration. Adolescent caffeine consumption also produced an upward shift in the U-shaped dose response curve on cocaine self-administration maintained on a within-session dose-response procedure. Adolescent caffeine consumption had no effect on cocaine seeking during extinction training or reinstatement of cocaine seeking by cues or cocaine. These findings suggest that caffeine consumption during adolescence may enhance the reinforcing properties of cocaine, leading to enhanced acquisition that may contribute to increased addiction vulnerability.


A study of histopathologic spectrum of nodular hidradenoma.
Hidradenomas are rare benign adnexal neoplasms that encompass a morphological gamut with a range of differentiation. As a consequence, there is a great likelihood of being mistaken for other primary and metastatic tumors. Though conventionally regarded as eccrine, they have been reclassified into eccrine and apocrine types. This study aims to document the histological spectrum of nodular hidradenomas, with particular reference to categorizing them into eccrine or apocrine tumors. A total of 15 cases with features of nodular hidradenoma with their age ranging from 18 years to 73 years were studied. Most of the cases were solitary, circumscribed, solid and cystic, dermal, symmetrical, lobulated tumors with a sheet-like and papillary architecture. The cells were chiefly eosinophilic with a regular oval grooved nucleus and a small inconspicuous nucleolus. Clear cells were also seen. Squamous differentiation was an important feature, with most showing a infundibular type of keratinization. Sebaceous differentiation is also common. The stroma varied from hyaline to myxoid. Only 1 case showed poroid differentiation. This study describes the assortment of histologic characteristics in hidradenomas. Apocrine hidradenomas are more common, contrary to earlier belief that favored an eccrine origin.


Expansion and fibronectin-enhanced retroviral transduction of primary human T lymphocytes for adoptive immunotherapy.
Human lymphocytes remain among the most promising target cells for gene therapy. Gene-modified lymphocytes have been used successfully to treat adenosine deaminase (ADA)-deficient patients and to control GvHD after allogeneic BMT. Because activation and proliferation of T cells are necessary for efficient retrovirus-mediated gene transfer and subsequent selection of transduced cells, mononuclear cells (MNC) from steady-state and G-CSF-stimulated peripheral blood were activated by short exposure to the mitogen PHA, the anti-CD3 antibody OKT3, or both in the presence of different concentrations of recombinant IL-2. Using OKT3 (10 or 30 ng/ml) and IL-2 (100 U/ml), T cells expanded efficiently during a 14-day culture period. Cell expansion was similar under serum-free conditions. The immunophenotypic profile over time showed a marked increase in CD8+ cells, leading to a reversed CD4/CD8 ratio of 1:2 and a slight increase in CD56+ cells. Supernatant-based centrifugal transduction of primary human T lymphocytes was compared with supernatant transduction on the extracellular matrix protein fibronectin. Transduction with cell-free retrovirus-containing supernatant in tissue culture flasks coated with human plasma fibronectin led to significantly higher transduction efficiencies (20% +/- 7.5%) than centrifugal transduction in uncoated culture flasks (13.6% +/- 5.1%)(p = 0.041). To both rapidly characterize transduced cells and isolate these from residual nontransduced but biologically equivalent cells, an amphotropic Moloney murine leukemia virus (MoMuLV)-based retroviral vector containing the intracytoplasmically truncated human low-affinity nerve growth factor receptor (deltaLNGFR) cDNA as a marker gene was used. FACS sorting of T cells after transduction resulted in >90% LNGFR+ cells and was much faster than enrichment of transduced cells through growth in G418-selection medium. These results show that supernatant-based retroviral gene transfer into primary human T lymphocytes can be enhanced by fibronectin. Ectopic expression of a cell surface protein can be used to rapidly and conveniently quantitate transduction efficiency through FACS analysis and to efficiently enrich transduced cells through FACS sorting.


Analysis of recombinant adenoviruses using an integrated microfluidic chip-based system.
The use of recombinant virus for gene therapy requires rigorous quality control methods to ensure that the viral vector preparations are functional and safe. A viral identity test is performed in which the viral payload, or transgene, is PCR amplified, followed by digestion with restriction enzymes that yields a characteristic "fingerprint." These DNA fragments are typically analyzed by agarose gel electrophoresis. The ethidium bromide-stained gels are photographed or scanned and the results are sufficient for a qualitative or semiquantitative identity confirmation of the viral product. We have investigated the use of an integrated microfluidic chip-based system as a new tool in the quality control testing of a recombinant, adenoviral, gene therapy product. The chip-based method was found to be very sensitive, requiring 100-fold less sample and only one-third the time compared to the agarose gel method. The automated data analysis sizes and quantitates the DNA fragments, thus yielding a more thorough, reproducible, sensitive, and rapid analysis.


Paclitaxel: severe mucocutaneous toxicity in a patient with hyperbilirubinemia.
Paclitaxel is a novel anticancer drug that is being increasingly used to treat cancer of the breast and other organs. We describe a patient with metastatic breast cancer and liver dysfunction who had severe mucocutaneous toxicity after administration of a standard dose of paclitaxel. Another interesting finding in this patient was that the administration of paclitaxel led to a prompt resolution of parathyroid hormone-related protein (PTHrP)-mediated hypercalcemia, which had previously proven to be refractory to multiple conventional antihypercalcemic agents as well as anthracycline-containing chemotherapy combination. The need for definitive guidelines for paclitaxel administration in the setting of hepatic dysfunction and the potentially unique sensitivity of PTHrP--producing cells to paclitaxel are discussed.


Orthographic analogies and phonological priming: a comment on Bowey, Vaughan, and Hansen (1998)
J. A. Bowey, L. Vaughan, and J. Hansen (1998, Journal of Experimental Child Psychology, 68, 108-133) carried out two experiments on 6- and 7-year-old children's use of orthographic analogies in word reading. They reported that, following apparently stringent controls for phonological priming effects, beginning analogies (beak-bean) were more frequent in this age group than rime (beak-peak) analogies. From this, they concluded that beginning readers do not reliably use orthographic rimes in reading, even in the clue word task (p. 129). However, the clue word task was not used in this study. This comment highlights two problems with Bowey et al.'s paper. The first is a theoretical one, and the second is methodological. Firstly, Bowey et al. base their investigation on a misunderstanding of U. Goswami and P. E. Bryant's (1990, Phonological skills and learning to read, Hillsdale, NJ: Erlbaum) claims about the role of rhyme and analogy in beginning reading. Secondly, methodological weaknesses, in particular unintended intralist priming effects, seriously limit the conclusions that can be drawn from Bowey et al.'s booklet analogy task.


Modification of Kaolinite Surfaces through Intercalation with Potassium Acetate, II.
The modification of kaolinite hydroxyl surfaces of both ordered and disordered kaolinites has been studied under a range of conditions and intercalating agents. This has been achieved by the study of the kaolinite hydroxyl deformation vibrations by the application of diffuse reflectance Fourier transform infrared (DRIFT) spectroscopy. Kaolinites showed resolved bands at 940 and 915 cm-1 with an additional band at 923 cm-1. The bands at 940 and 923 cm-1 are assigned to the hydroxyl deformation modes of the inner surface hydroxyl groups, and the 915 cm-1 band is assigned to that of the inner hydroxyl group. When the kaolinite is intercalated with potassium acetate additional bands were found at 909 and 897 cm-1 for the ordered kaolinites. An additional band at 877 cm-1 is observed for the disordered kaolinite and is attributed to the hydroxyl deformation mode of non-hydrogen-bonded hydroxyl groups. The bands at 909 and 897 cm-1 range are attributed to inner surface hydroxyl groups hydrogen bonded to water or acetate groups. The intensity of these bands reflects the degree of disorder introduced into the kaolinite through intercalation. The intensities of the kaolinite hydroxyl deformation modes are a function of the starting kaolinite order and the method of preparation. Copyright 1999 Academic Press.


Lung cancer screening in high-risk populations.
Although the number of smokers per capita has declined appreciably in the United States in the past 30 years, smokers still make up about one quarter of the adult population. It does not appear that the number of US smokers will decrease further in the next century, and the number may even increase due to the popularity of smoking among teenagers. Epidemiological data indicate that women are more susceptible, dose-for-dose, to the adverse effects of tobacco smoke. Since women make up a large percentage of today s smokers, lung cancer rates may increase in the future. Current guidelines recommend against lung cancer screening based on chest x-ray and sputum morphology; however, new highly sensitive detection methods are available that may make screening more effective, especially if combined with analysis of risk factors for lung cancer and biomarkers of damage to the airways that may identify individuals at highest risk for lung malignancies. Lung cancer will continue to be a major public health problem in the next century. Advances in the field of early detection may make lung cancer screening practical and effective in the near future.


Derepression and Repression of Lysine-sensitive Aspartokinase during in Vitro Culture of Carrot Root Tissue.
The increment of lysine-sensitive aspartokinase (EC 22.214.171.124) activity during in vitro culture of carrot (Daucus carota, cv. Oogata sanzun) root tissue was explained in terms of derepression caused by an earlier decrease in the endogenous level of lysine, a possible end product repressor. Tissue content of free lysine decreased to about one-third of the initial level after 1 day of culture and no lysine was detected in the 2nd day. Inclusion of lysine (0.1 to 1.0 millimolar) in the culture medium resulted in a specific suppression of increase in lysine-sensitive aspartokinase activity without affecting the increase in threonine-sensitive aspartokinase activity.


Strong aggregation and increased toxicity of polyleucine over polyglutamine stretches in mammalian cells.
Expansion of a Glutamine (Gln) repeat above a specific critical size in certain proteins gives rise to aggregation-prone proteins that cause neurodegenerative disorders, such as Huntington's disease. However, proteins with long hydrophilic polyglutamine repeats are more frequently found in nature than proteins with long homogeneous repeats of other amino acids, such as hydrophobic (Ala)(n) and (Leu)(n). To explore this finding, the effects of expression in mammalian cells of polyglutamine and polyleucine encoded by mixed DNA repeats were compared. It was found that polyleucine is significantly more toxic than polyglutamine. In addition, we show that polyleucine stretches display a high propensity for aggregation utilizing two complementary biochemical assays and that polyleucine stretches can also be detected by the monoclonal antibody 1C2, which specifically recognizes expanded pathogenic and aggregation-prone glutamine repeats. Together, these results suggest that polyglutamine stretches are in fact relatively well tolerated and that nature may select more strongly against DNA stretches that encode long hydrophobic homopolymeric amino acid stretches, such as polyleucine -- possibly owing to their strong propensity for aggregation. In keeping with this notion, an increasing number of diseases are found to be associated with expansion of stretches of hydrophobic amino acids, including oculopharyngeal muscular dystrophy (OPMD), which is associated with expansion of a hydrophobic polyalanine stretch.


The increasingly complex mechanism of HMG-CoA reductase.
HMG-CoA reductase (HMGR) is the target of statins, cholesterol-lowering drugs prescribed to millions of patients worldwide. More recent research indicates that HMGR could be a useful target in the development of antimicrobial agents. Over the last seven decades, researchers have proposed a series of increasingly complex reaction mechanisms for this biomedically important enzyme. The maturation of the mechanistic proposals for HMGR have paralleled advances in a diverse set of research areas, such as molecular biology and computational chemistry. Thus, the development of the HMGR mechanism provides a useful case study for following the advances in state-of-the-art methods in enzyme mechanism research. Similarly, the questions raised by these mechanism proposals reflect the limitations of the methods used to develop them. The mechanism of HMGR, a four-electron oxidoreductase, is unique and far more complex than originally thought. The reaction contains multiple chemical steps, coupled to large-scale domain motions of the homodimeric enzyme. The first proposals for the HMGR mechanism were based on kinetic and labeling experiments, drawing analogies to the mechanism of known dehydrogenases. Advances in molecular biology and bioinformatics enabled researchers to use site-directed mutagenesis experiments and protein sequencing to identify catalytically important glutamate, aspartate, and histidine residues. These studies, in turn, have generated new and more complicated mechanistic proposals. With the development of protein crystallography, researchers solved HMGR crystal structures to reveal an unexpected lysine residue at the center of the active site. The many crystal structures of HMGR led to increasingly complex mechanistic proposals, but the inherent limitations of the protein crystallography left a number of questions unresolved. For example, the protonation state of the glutamate residue within the active site cannot be clearly determined from the crystal structure. The differing protonation state of this residue leads to different proposed mechanisms for the enzyme. As computational analysis of large biomolecules has become more feasible, the application of methods such as hybrid quantum mechanics/molecular mechanics (QM/MM) calculations to the HMGR mechanism have led to the most detailed mechanistic proposal yet. As these methodologies continue to improve, they prove to be very powerful for the study of enzyme mechanisms in conjunction with protein crystallography. Nevertheless, even the most current mechanistic proposal for HMGR remains incomplete due to limitations of the current computational methodologies. Thus, HMGR serves as a model for how the combination of increasingly sophisticated experimental and computational methods can elucidate very complex enzyme mechanisms.


Modeling of Photochemical Reactions in a Focused Laser Beam.
Fluorescent materials play a prominent role in the qualitative and quantitative measurement of scientific phenomena of importance in biotechnology and biomedical applications. Photodegradation of fluorophores is a process that determines the accuracy and sensitivity of such measurements. This is the motivation for developing methods for accurately measuring fluorophore photodegradation rates. Recently, illumination consisting of short pulses has been used to examine the decay of photochemical reaction products. However, the time resolved measurements are difficult to interpret since the photodegradation process usually involves multiple time scales. The frequency domain measurement technique discussed here looks at the frequency response of a fluorescent sample to a frequency modulated illuminating light. The photodegradation rate is obtained by interpreting the frequency domain measurements in terms of traditional impedance concepts. In the measurements described in this paper, a focused laser beam is used to illuminate a sample of slowly flowing fluorescent solution. The laser beam is assumed to have a Gaussian power distribution hence illumination is spatially non-uniform in the region of interest. The photochemical reaction rates depend on power, so they will also vary with the position in the beam. However in the case of photodegradation of fluorophores, the measurement of the resulting decrease in fluorescence is given in terms of the radiation emitted from the entire illuminated region. In this work we present a mathematical description of the time evolution of the fluorescence response integrated over a non-uniformly illuminated domain. As a result of our analysis, an experimentally accessible and tractable mathematical model Eq. (19) and Eq. (30) is obtained from a more fundamental description given by Eq. (4) and Eq. (5). The model is used to create a functional form for fitting experimental measurements from a lock-in amplifier.


The role of bacterial adherence in otitis media with effusion.
Adherence of nontypable Haemophilus influenzae and Streptococcus pneumoniae to nasopharyngeal epithelial cells was investigated in vitro. Both strains had higher affinity to the epithelial cells of children than to those of adults. In children, the adherence was significantly greater in patients with otitis media with effusion than in normal subjects. Secretory IgA in nasopharyngeal secretions was found to have antibody activity against the bacteria. Adherence of both bacteria was significantly smaller in the group having secretory IgA antibody activity than in the group having no activity. These results suggest that bacterial adherence to the nasopharynx may play an important role in the pathogenesis of otitis media with effusion in children, and that secretory IgA in nasopharyngeal secretions may be related to the decrease of adherence.


High connectivity in the deepwater snapper Pristipomoides filamentosus (Lutjanidae) across the Indo-Pacific with isolation of the Hawaiian archipelago.
In the tropical Indo-Pacific, most phylogeographic studies have focused on the shallow-water taxa that inhabit reefs to approximately 30 m depth. Little is known about the large predatory fishes, primarily snappers (subfamily Etelinae) and groupers (subfamily Epinephelinae) that occur at 100-400 m. These long-lived, slow-growing species support fisheries across the Indo-Pacific, yet no comprehensive genetic surveys within this group have been conducted. Here we contribute the first range-wide survey of a deepwater Indo-Pacific snapper, Pristipomoides filamentosus, with special focus on Hawai'i. We applied mtDNA cytochrome b and 11 microsatellite loci to 26 samples (N=1,222) collected across 17,000 km from Hawai'i to the western Indian Ocean. Results indicate that P. filamentosus is a highly dispersive species with low but significant population structure (mtDNA Φ(ST)=0.029, microsatellite F(ST)=0.029) due entirely to the isolation of Hawai'i. No population structure was detected across 14,000 km of the Indo-Pacific from Tonga in the Central Pacific to the Seychelles in the western Indian Ocean, a pattern rarely observed in reef species. Despite a long pelagic phase (60-180 days), interisland dispersal as adults, and extensive gene flow across the Indo-Pacific, P. filamentosus is unable to maintain population connectivity with Hawai'i. Coalescent analyses indicate that P. filamentosus may have colonized Hawai'i 26 K-52 K y ago against prevailing currents, with dispersal away from Hawai'i dominating migration estimates. P. filamentosus harbors low genetic diversity in Hawai'i, a common pattern in marine fishes, and our data indicate a single archipelago-wide stock. However, like the Hawaiian Grouper, Hyporthodus quernus, this snapper had several significant pairwise comparisons (F(ST)) clustered around the middle of the archipelago (St. Rogatien, Brooks Banks, Gardner) indicating that this region may be isolated or (more likely) receives input from Johnston Atoll to the south.


Cloning and sequence analysis of a plasmid-encoded chloramphenicol acetyltransferase gene from Staphylococcus intermedius.
The chloramphenicol acetyltransferase gene (cat) of a 3.9 kb chloramphenicol resistance (CmR) plasmid from Staphylococcus intermedius, designated pSCS1, was cloned into an Escherichia coli plasmid vector. Sequence analysis revealed a high degree of base similarity with the cat gene of the S. aureus CmR plasmid pC221 but there were several differences in the regulatory region. A lesser degree of similarity was observed between the cat gene of the S. intermedius plasmid and the cat gene of the S. aureus plasmid pC194.


[The Swiss Health Leagues Conference].
The Conference of Swiss Health Leagues-founded in 1971-unites associations which aim at prevention and control of chronic diseases and their physical and social consequences. The semi-annual conferences, the commissions and working parties enable exchange of experience, coordination of activities, and the pursuing of common interests in very specialized and heterogenous member organizations.


Thirteen-month duration of immunity of an oral canine vaccine against challenge with Bordetella bronchiseptica.
Very few studies have evaluated the duration of immunity of Bordetella bronchiseptica vaccines in dogs, and to date, no studies have been published on the duration of immunity of oral canine Bordetella bronchiseptica vaccines. This study was designed to determine the effectiveness of a single dose of an oral B bronchiseptica vaccine in dogs when challenged 13 months after vaccination. Two groups of approximately eight-week-old beagles were vaccinated once with 1 ml of placebo vaccine (oral, n=17) or 1 ml of Recombitek Oral Bordetella (oral, n=17). Thirteen months after vaccination, both groups were challenged with virulent B bronchiseptica via aerosolisation. Thirteen of 17 dogs in the placebo group (76.5 per cent) and no dogs in the Recombitek Oral Bordetella vaccine group (0.0 per cent) developed spontaneous cough of two or more consecutive days (disease case definition). Dogs in the Recombitek Oral Bordetella group had a significantly lower prevalence of disease with prevented fraction of 1 (100 per cent prevention). In addition, the number of days coughing, duration of cough and prevalence of tracheal and nasal shedding were significantly lower for dogs vaccinated with Recombitek Oral Bordetella. The study demonstrated that vaccination with Recombitek Oral Bordetella is effective in preventing disease and reducing shedding 13 months after vaccination when compared with dogs vaccinated with a placebo.


Optical resonator biosensors: molecular diagnostic and nanoparticle detection on an integrated platform.
Optical resonator biosensors are emerging as one of the most sensitive microsystem biodetection technology that does not require amplification or labeling of the analyte. This minireview provides a scholarly introduction to this research area and reviews current advances in molecular diagnostics and nanoparticle detection.


[Free-radical lipid oxidation in rabbits with the alimentary peroxidation syndrome].
The results are presented of the study of free radical oxidation of lipids in rabbits with alimentary peroxidation syndrome. A long-term feeding of the animals with a ration devoid of alimentary antioxidants reproduces the main biochemical changes in the blood and tissues characteristic of old animals. It has been revealed that the intensity of EPR signals of lyophilized tissues increased proportionally to the animals' age and to the duration of their maintenance on the ration devoid of antioxidants. A conclusion has been made that the deprivation of the rabbits of alimentary antioxidants accelerates aging.


Level of superoxide dismutase, glutathione peroxidase and uric acid in thioacetamide-induced cirrhotic rats.
Levels of superoxide dismutase and glutathione peroxidase were determined in blood and hepatic tissues of thioacetamide-induced cirrhotic rats and compared to levels in age-matched control animals. The plasma level of uric acid was also determined in these animals. A general decrease was noticed in the level of all the antioxidants examined as compared to the control. This decrease was statistically significant in the level of all the antioxidants studied, except for the level of superoxide dismutase in blood. A decrease in the antioxidant level may indicate an increase in free radical level and thereby an increase in cellular damage in cirrhotic rats. The changes in the level of antioxidants showed a direct correlation with the changes in the level of trace elements observed in our previous studies. These studies suggest that antioxidants alone or in combination with trace elements may have beneficial effects in treating liver cirrhosis.


[Conformation of a DNA molecule in water-salt solutions depends on the salt anion type].
The methods of viscosimetry and flow birefringence were used to study the effect of anions F-, Cl-, Br-, J-, NO3-, ClO4-, SCN-, CH3COO- on the dimensions and thermodynamic rigidity of the DNA molecule in solutions in a wide range of ionic strengths and at different temperatures. It was shown that the persistent lengtH of DNA is independent of the anion type at all values of ionic strength, and the changes in its dimensions, which are determined from the changes in intrinsic viscosity, are due to the influence of anions and their hydration on long-range interactions in the macromolecule. Possible reasons for this phenomena and the role of structural changes in water upon hydration of anions are discussed.


Non-uniform velocity of homogeneous DNA in a uniform electric field: consequence of electric-field-induced slow dissociation of highly stable DNA-counterion complexes.
Identical molecules move with identical velocities when placed in a uniform electric field within a uniform electrolyte. Here we report that homogeneous DNA does not obey this fundamental rule. While most DNA moves with similar velocities, a fraction of DNA moves with velocities that vary within a multiple-fold range. The size of this irregular fraction increases several orders of magnitude when exogenous counterions are added to DNA. The irregular fraction decreases several orders of magnitude when DNA counterions are removed by dialysis against deionized water in the presence of a strong electric field (0.6 kV/cm). Dialysis without the field is ineffective in decreasing the size of irregular fraction. These results suggest that (i) DNA can form very stable complexes with counterions, (ii) these complexes can be dissociated by an electric field, and (iii) the observed non-uniform velocity of DNA is caused by electric-field-induced slow dissociation of these stable complexes. Our findings help to better understand a fundamental property of DNA: its interaction with counterions. In addition, these findings suggest a practical way of making electromigration of DNA more uniform: removal of strongly bound DNA counterions by electro-dialysis against deionized water.


Epstein-Barr virus BNRF1 destabilizes SMC5/6 cohesin complexes to evade its restriction of replication compartments.
Epstein-Barr virus (EBV) persistently infects people worldwide. Delivery of ∼170-kb EBV genomes to nuclei and use of nuclear membrane-less replication compartments (RCs) for their lytic cycle amplification necessitate evasion of intrinsic antiviral responses. Proteomics analysis indicates that, upon B cell infection or lytic reactivation, EBV depletes the cohesin SMC5/6, which has major roles in chromosome maintenance and DNA damage repair. The major tegument protein BNRF1 targets SMC5/6 complexes by a ubiquitin proteasome pathway dependent on calpain proteolysis and Cullin-7. In the absence of BNRF1, SMC5/6 associates with R-loop structures, including at the viral lytic origin of replication, and interferes with RC formation and encapsidation. CRISPR analysis identifies RC restriction roles of SMC5/6 components involved in DNA entrapment and SUMOylation. Our study highlights SMC5/6 as an intrinsic immune sensor and restriction factor for a human herpesvirus RC and has implications for the pathogenesis of EBV-associated cancers.


Characterisation of immune responses to food allergens in mice.
There is considerable interest in the development and evaluation of approaches for the safety assessment of novel foods, and in particular in methods for characterisation of allergenic potential. One strategy that has found favour is a tiered approach in which the potential of novel proteins to induce allergic sensitisation is assessed based on considerations of stability of the protein in a simulated gastric juice and homology with, or structural similarity to, known allergens. Linked to such an approach may be evaluation of serological identity with proteins known to cause allergic disease. With the aim of supplementing such approaches with a more direct measurement of potential allergenic activity, attempts have been made to characterise the quality of immune responses elicited in BALB/c strain mice. Such evaluations comprise measurement of IgG and IgE antibody production and (to a lesser extent) of induced cytokine expression patterns. Investigations to date suggest that in mice proteins provoke variable immune responses, those with the potential to cause allergic sensitisation stimulating IgE (and IgG) antibody production. In contrast, non-allergenic, but nevertheless immunogenic, proteins are associated with IgG antibody responses in the absence of marked IgE production. Consistent with the selective activation of selective type 2 T lymphocyte responses, exposure of mice to allergenic protein is associated with preferential expression of IL-4, -5, -10 and -13. Collectively these data suggest that characterisation of the nature of immune response induced in mice by proteins may provide a useful adjunct or alternative to current strategies for the assessment of allergenic potential.


Organizational complexity of a rice transgene locus susceptible to methylation-based silencing.
Molecular analyses of a rice (Oryza sativa L.) transgene locus introduced using biolistic techniques revealed the presence of multiple copies of rearranged fragments, as well as an intact copy of the supplied constructs. Both the gene of interest (35S-Btt cryIIIA) and the selectable marker used (Ubi1-bar) were methylated and silenced. Additionally, vector sequences were present in great abundance and were also highly methylated, indicating that the entire transgene insert was marked for methylation. The rearrangement of input DNA resulted in interspersion of plasmid backbone regions with the gene of interest. Permutation of segments encoding the gene of interest and the selectable marker was also detected, perhaps explaining why sequences introduced on separate plasmids are frequently found to be inserted at the same locus. The 35S promoter contained several hotspots for fragmentation. These observations strongly support the concept that intrusive DNA is recognized by host surveillance systems and that transgene loci with anomalous structural organization are subjected to inactivation by processes such as methylation.


Studies on human pregnancy-induced insulin-like growth factor (IGF)-binding protein-4 proteases in serum: determination of IGF-II dependency and localization of cleavage site.
Insulin-like growth factor (IGF)-binding protein-4 (IGFBP-4), a consistent inhibitor of IGF action, is subject to proteolytic cleavage by the IGF-II-dependent IGFBP-4 protease. However, regulation of the IGF-II-dependent IGFBP-4 protease in vivo is not known. As IGFBP proteases are known to be triggered during pregnancy, we systematically evaluated the changes in IGFBP-4 proteolysis by serum collected throughout human pregnancy. Results from in vitro protease assays using recombinant IGFBP-4 revealed that IGFBP-4 proteolysis determined in both the presence and absence of exogenous IGF-II significantly increased during the first and second trimesters and reached a plateau by the third trimester. However, in the absence of IGF-II, IGFBP-4 proteolysis by pregnancy serum was only observed after prolonged incubation. IGF-II dose dependently increased IGFBP-4 proteolysis by pregnancy serum, with maximal stimulation observed at a concentration of 0.7 mol/L relative to IGFBP-4. In contrast, IGF-II at an equimolar dose had little effect on proteolysis of recombinant human IGFBP-3, whereas excess IGF-II reproducibly inhibited recombinant human IGFBP-3 proteolysis by pregnancy serum. Although IGF-II enhanced IGFBP-4 proteolysis, results from N-terminal sequence and mass spectrometric analyses of IGFBP-4 proteolytic fragments demonstrate that the cleavage site (Met135-Lys136) in human IGFBP-4 was not altered by IGF-II. Deletion of the residues 121-141 containing this cleavage site blocked IGFBP-4 proteolysis. These findings demonstrate that the increase in IGFBP-4 proteolysis during pregnancy was accounted for mainly by the IGF-II-dependent IGFBP-4 proteolysis. Because IGFBP-4 is a potent inhibitor of IGF actions, it can be speculated that the pregnancy-induced IGFBP-4 proteases may play an important role in regulating fetal growth.


Assessment of fed-batch cultivation strategies for an inducible CHO cell line.
In order to maximize cell growth and productivity for an inducible CHO cell line expressing rituximab, various fed-batch culture strategies were investigated. In each case, the performance was evaluated for cultures induced at moderate and high cell density conditions (4 × 106 and 10 × 106 cells/mL) to assess the impact of the timing of induction. We first demonstrate the importance of starting the feeding process during the growth phase, as this translated into significantly improved integral of viable cells and antibody concentration, when compared to post-induction feeding only. Secondly, we investigated the impact of the feed rate by maintaining different levels of glucose (25, 35 and 50 mM) via a dynamic feeding strategy. The highest antibody concentrations were achieved under a moderate feeding regime for both cell densities at induction, highlighting the risks of under- or over-feeding the cultures. We then evaluated the impact of performing a temperature shift at induction by testing different mild hypothermia conditions. At small-scale, the highest production yields (1.2 g/L) were achieved when the temperature was reduced from 37 to 30 °C during the production phase of a culture induced at high cell density. When the strategy was applied in bioreactor, the better controlled conditions led to even greater product concentrations (1.8 g/L). Furthermore, this production protocol was shown to promote a more galactosylated glycan profile than a bioreactor culture initiated at 34 °C during growth and downshifted to 30 °C during the production phase.


Internal representation of simple temporal patterns.
In this study the imitation of several periodically repeating simple temporal patterns consisting of two or more intervals varying in their duration ratios has been investigated. The errors that subjects typically made in their imitations and the systematic changes that occurred during repeated imitations indicate that both musically trained and untrained subjects map temporal sequences onto an interval structure the nature of which is revealed by studying which patterns are correctly and which incorrectly reproduced. A "beat-based" model for the perception of temporal sequences is proposed. This model states that the first step in the processing of a temporal sequence consists of a segmentation of the sequence into equal intervals bordered by events. This interval is called the beat interval. How listeners select this beat interval is only partly understood. In a second step, intervals smaller than the beat interval are expressed as a subdivision of the beat interval in which they occur. The number of within-beat structures that can be represented in the model is, however, limited. Specifically, only beat intervals that are subdivided into either equal intervals or intervals in a 1:2 ratio fit within the model. The partially hierarchical model proposed, though in need of further elaborations, shows why the number of temporal patterns that can be correctly conceptualized is limited. The relation of the model to other models is discussed.


Expression of regeneration-associated cytoskeletal proteins reveals differences and similarities between regenerating organs.
The unique events which allow regeneration of an entire organ to occur are formation of a specialized wound epidermis and accumulation of progenitor cells (blastemal cells) at the amputated surface to form a blastema. In order to identify some of the molecular events underlying the early stages of the regenerative process which are either common to different systems or specific to one of them, we have investigated whether molecules which are induced in limb blastemas are also expressed in skin repair and during regeneration of other complex body structures (lower jaws, upper jaws, and tails). In addition, we have addressed the issue of the identity of progenitor cells during jaw development and regeneration by analyzing the expression of limb blastemal markers in the developing head and face. We have focused on cytoskeletal components, and particularly on the epidermal keratin NvKII, the simple epithelial keratins 8 and 18 and 22/18, because they are among the few molecules which have been shown to be associated with regeneration in the limb and may play significant roles in various developmental processes. Some important findings emerge from this study: 1) Expression of the epidermal keratin NvKII, unlike that of its mammalian homologue K6, is not simply induced in response to wounding, but is associated with regeneration of specific organs. In fact, NvKII is expressed in regenerating limbs and tails, but not in upper or in lower jaw regenerates, demonstrating the existence of molecular differences in the composition of the wound epidermis in these systems. This, together with the fact that NvKII mRNA is regulated by retinoic acid, which differentially affects patterning of limbs and jaws, argues for distinct inductive abilities of the wound epidermis in different organs. 2) In contrast to the differential expression of the epidermal keratin NvKII, the regeneration-associated cytoskeletal molecules identified in limb blastemal cells are expressed in a similar fashion in jaw and tail blastemas. Therefore, it appears that similar cellular events lead to the establishment of an actively proliferating population of progenitor cells from the stump of different organs. Finally, the mesenchyme of the facial rudiments, unlike that of developing limb buds, expresses simple epithelial keratins. Thus, it appears that mesenchymal progenitor cells of developing and regenerating jaws are alike in regard to their intermediate filament content, and this may be related to nerve-dependent growth control of progenitor cells in different developing and regenerating systems.


[Research in basic science for health protection of coal miners in Kuzbass].
The article covers results of studies in occupational hygiene and physiology, occupational traumatism, morbidity among coal miners of Kouzbass; evaluation of occupational morbidity risks and environmental state in coal miner towns.


pH-Mediated Antibacterial Dyeing of Cotton with Prodigiosins Nanomicelles Produced by Microbial Fermentation.
This study developed a novel pH-mediated antimicrobial dyeing process of cotton with prodigiosins nanomicelles produced by microbial fermentation. The average diameter of the pigment nanomicelles was 223.8 nm (range of 92.4⁻510.2 nm), and the pigment concentration was 76.46 mg/L. It was found that the superior dyeing effect of cotton fabric was achieved by adjusting the dye bath pH. When the pH was three, dyed cotton under 90 °C for 60 min exhibited the greatest color strength with good rubbing, washing and perspiration color fastness. By the breaking strength test and XRD analysis, it was concluded that the cotton dyed under the optimum condition almost suffered no damage. In addition, due to the presence of prodigiosins, dyed cotton fabric under the optimal process showed outstanding bacteriostatic rates of 99.2% and 85.5% against Staphylococcus aureus and Escherichia coli, respectively. This research provided an eco-friendly and widely-applicable approach for antimicrobial intracellular pigments with the property of pH-sensitive solubility in water to endow cellulose fabric with color and antibacterial activity.


Studies of the fragmentation of the monocation and dication of methanol.
Relative partial ionization cross sections and precursor-specific relative partial ionization cross sections for fragment ions formed by electron ionization of methanol have been measured using time-of-flight mass spectrometry coupled with a two-dimensional ion coincidence technique. Relative cross sections are reported for ionizing energies from 30 to 200 eV. Good agreement is found between our data and one set of recently published absolute partial ionization cross sections. Conversely, discrepancies are observed with another set of recently published data; we attribute these discrepancies to the loss of translationally energetic fragment ions. Our precursor-specific cross sections allow the contribution from single and double ionization to the individual fragment ion yields, following ionization of methanol, to be quantified for the first time. Our analysis shows that the contribution of double ionization to the total ion yield reaches a maximum of 20% between 150 and 200 eV.


Optimized Hydrogen Mass Repartitioning Scheme Combined with Accurate Temperature/Pressure Evaluations for Thermodynamic and Kinetic Properties of Biological Systems.
In recent years, molecular dynamics (MD) simulations with larger time steps have been performed with the hydrogen-mass-repartitioning (HMR) scheme, where the mass of each hydrogen atom is increased to reduce high-frequency motion while the mass of a non-hydrogen atom bonded to a hydrogen atom is decreased to keep the total molecular mass unchanged. Here, we optimize the scaling factors in HMR and combine them with previously developed accurate temperature/pressure evaluations. The heterogeneous HMR scaling factors are useful to avoid the structural instability of amino acid residues having a five- or six-membered ring in MD simulations with larger time steps. It also reproduces kinetic properties, namely translational and rotational diffusions, if the HMR scaling factors are applied to only solute molecules. The integration scheme is tested for biological systems that include soluble/membrane proteins and lipid bilayers for about 200 μs MD simulations in total and give consistent results in MD simulations with both a small time step of 2.0 fs and a large, multiple time step integration with time steps of 3.5 fs (for fast motions) and 7.0 fs (for slower motions). We also confirm that the multiple time step integration scheme used in this study provides more accurate energy conservations than the RESPA/C1 and is comparable to the RESPA/C2 in NAMD. In summary, the current integration scheme combining the optimized HMR with accurate temperature/pressure evaluations can provide stable and reliable MD trajectories with a larger time step, which are computationally more than 2-fold efficient compared to the conventional methods.


A comparative trial of a single dose of azithromycin versus doxycycline for the treatment of mild scrub typhus.
Scrub typhus is one of the most important endemic infections in the Asia-Pacific region. Although tetracyclines or chloramphenicol are the recommended drugs of choice for the treatment of scrub typhus, reports of doxycycline-resistant strains have prompted a search for alternative treatments. We conducted a prospective, open-label, randomized trial from September 2002 through November 2003 to compare azithromycin with doxycycline for the treatment of mild scrub typhus. The time to defervescence was assessed to compare the efficacy of the 2 treatment regimens. A total of 93 patients were randomly assigned to receive either a single 500-mg dose of azithromycin or a 1-week course of daily oral 200-mg dose of doxycycline. Cure was achieved in 47 (100%) of 47 patients in the azithromycin-treated group and in 43 (93.5%) of 46 patients in the doxycycline-treated group (P=.117). The median time to defervescence was 21 h for the azithromycin-treated group and 29 h for the doxycycline-treated group (P=.097). There were no serious adverse events during the study. No relapses occurred in either group during a 1-month follow-up period. The single 500-mg dose of azithromycin was as effective as the 1-week course of daily 200-mg doses of doxycycline for the treatment of mild scrub typhus acquired in South Korea.


[Management of advertising communication in health care organizations].
The article examines the impact of advertising communications on the patient flow of medical organizations in the Russian healthcare system, reflecting significant changes in the global communication industry. The study of the current changes indicates the relevance of the topic of marketing communications among the heads of Russian and foreign medical companies, government agencies responsible for medicine, the media, advertising companies and consumers of medical services. Issues of concern to the medical services advertising industry and regulatory authorities in healthcare, the impact of advertising on the development of the medical market, and international experience in state regulation of advertising communications in the field of medicine are relevant. Today, the development of an effective policy aimed at attracting new customers and expanding the service sector is a prerequisite for the success of any medical organization. Along with the increasing competition of organizations working in the medical market for consumers of medical services, the need for effective management of advertising communications is also increasing, thanks to which the company's status is maintained, qualified, high-quality treatment is provided, and a high level of service is maintained. Thus, the management of advertising communications in medical organizations has its own specifics and requires compliance with Russian legislation. Ads for medical services must truthfully inform the patient about the nature and direction of the medical organization's work.


Role of intrapartum antibiotics in prevention of vertical transmission of neonatal sepsis.
The present study evaluates the role of prophylactic intrapartum antibiotics in the prevention of neonatal sepsis. Labour and delivery characteristics of 1478 women delivering at the Nehru Hospital, PGIMER, Chandigarh were recorded. Intrapartum antibiotics (ampicillin, with or without gentamycin) were given to 69 per cent women. The neonatal sepsis rate was 1.56 per cent. This was not significantly lower in the women who received intrapartum antibiotics (1.47 vs 1.75%). Though there was a lowering of neonatal sepsis rate with intrapartum antibiotic administration when the duration of labour was more than 12 h (1.67 vs 3.09%), duration of ruptured membranes was more than 6 h (1.93 vs 3.81%) and number of pelvic examination was 3 or more (1.63 vs 4.54%), it was not statistically significant. It was concluded that intrapartum antibiotics as per the existing protocol did not prevent neonatal sepsis.


Biorecognition in Organic Field Effect Transistors Biosensors: The Role of the Density of States of the Organic Semiconductor.
Biorecognition is a central event in biological processes in the living systems that is also widely exploited in technological and health applications. We demonstrate that the Electrolyte Gated Organic Field Effect Transistor (EGOFET) is an ultrasensitive and specific device that allows us to quantitatively assess the thermodynamics of biomolecular recognition between a human antibody and its antigen, namely, the inflammatory cytokine TNFα at the solid/liquid interface. The EGOFET biosensor exhibits a superexponential response at TNFα concentration below 1 nM with a minimum detection level of 100 pM. The sensitivity of the device depends on the analyte concentration, reaching a maximum in the range of clinically relevant TNFα concentrations when the EGOFET is operated in the subthreshold regime. At concentrations greater than 1 nM the response scales linearly with the concentration. The sensitivity and the dynamic range are both modulated by the gate voltage. These results are explained by establishing the correlation between the sensitivity and the density of states (DOS) of the organic semiconductor. Then, the superexponential response arises from the energy-dependence of the tail of the DOS of the HOMO level. From the gate voltage-dependent response, we extract the binding constant, as well as the changes of the surface charge and the effective capacitance accompanying biorecognition at the electrode surface. Finally, we demonstrate the detection of TNFα in human-plasma derived samples as an example for point-of-care application.


The importance of computer observation of postoperative complications in ophthalmology presented on the model of postoperative complications following cataract surgery.
Medical documentation is getting more extensive, it is not standardised and can hardly be used to provide the exact and proper feedback. The crisis of data storage, processing and uninterrupted flow of information is becoming concial in the medical institutions as well. An example of informatic model of observation of postoperative complications following cataract surgery is presented in this work. The model has a very flexible pattern, and can, therefore, with minor changes, be used in observation of all postoperative complications in ophthalmology. It is based on the use of personal computer with professional software support (data base III programme was used for creation of data base, while statistical graphic analysis was carried out by following programmes: MICROSTAT, ABSTAT, STATGRAPHIC and FRAMEWORK). It has to be pointed out that any doctor, after only a short (optional) education, could prepare such models for his own purposes, and could thus assist in overcoming the information in medicine. In conclusion, the authors state computer is simple to work with, fast, economical and precise. Computerisation of health service shall include it in one of the aspects of practical realisation of the third technological revolution.


[Expression of Mn-SOD mRNA in streptozotocin-induced diabetic rat cornea by in situ hybridization].
The pathogenesis of diabetic corneal epitheliopathy, one of the ocular complications frequently seen in diabetes patients, still remains to be elucidated. Hyperglycemia causes glycation of various proteins leading to the formation of superoxide radicals (O2.-). Copper, zinc-superoxide dismutase (Cu, Zn-SOD), a scavenger of superoxide radicals, whose function is complementary to manganese-SOD (Mn-SOD), is inactivated during glycation. As a first step to clarify whether depressed antioxidant activity is associated with diabetic corneal epitheliopathy or not, we investigated the expression of Mn-SOD mRNA (messenger ribonuclic acid) in streptozotocin-induced diabetic rat cornea by in situ hybridization using a digoxigenin-labeled Mn-SOD cDNA probe. Mn-SOD mRNA was detected in epithelial cell layer and endothelial cell layer of both diabetic rat cornea and normal rat cornea. However, the expression of Mn-SOD mMRA in the epithelial cell layer of diabetic rat cornea was weaker than that of normal rat cornea. These results suggest that decreased Mn-SOD activity might be one of factors causing diabetic corneal epitheliopathy.


Radiation therapy for pelvic recurrent colorectal or gynecological cancer: is whole pelvic irradiation necessary?
Preoperative whole pelvic radiation therapy (RT) is used commonly for rectal cancer and is the standard field postoperatively in gynecological cancer. However, the ideal field (local vs. whole pelvis) has not been determined for local recurrence of these cancers. We retrospectively reviewed the data for 52 patients who developed local tumor recurrence of rectal or gynecological cancer treated from 2013 to 2021. The initial treatment for all patients was total excision of the primary tumors without radiation therapy. Radiation therapy targets were surgical stumps, perianastomosis sites, and pelvic lymph nodes, classified according to the pelvic nodal volume atlas for radiation therapy. Patients were divided into the local recurrent tumor only radiation therapy group and the whole pelvis radiation therapy group. Whole pelvis radiation therapy included the common iliac lymph nodes or prophylactic lymph nodes below the L5/S1 junction. We recorded second recurrence after RT and the affected site(s) in each group. We also compared disease-specific survival using uni- and multivariate analyses. We found no significant differences between the groups regarding second recurrence or regarding the site(s) of recurrence. We also found no significant differences in disease-specific survival between the two RT groups. However, patients who did not receive chemotherapy after the initial surgery and before RT had significantly longer survival (P=0.015). In patients with locally recurrent rectal or gynecological cancer, we found no significant difference in second recurrence or survival between the local tumor only RT field and the whole pelvic RT field.


Considerations regarding appropriate sample size for conducting ferret transmission experiments.
Evaluation of: Nishiura H, Yen H-L, Cowling BJ. Sample size considerations for one-to-one animal transmission studies of the influenza A viruses. PLoS ONE 8(1), e55358 (2013). There is an urgent need to model in a laboratory setting the capacity of wild-type influenza viruses to transmit between mammals, to determine the molecular determinants and identify biological properties that confer influenza virus transmissibility, and to explore both pharmaceutical and nonpharmaceutical methods to inhibit virus transmission. Owing to its close physiologic match to humans, researchers typically utilize the ferret to measure influenza virus transmissibility. Nishiura et al. highlight the dilemma facing researchers utilizing the ferret transmission model: how to provide high-quality data to guide public health efforts, while ensuring the ethical use of animals in limited-size, individual, one-to-one transmission experiments. However, the responsible interpretation of data generated using this model can overcome this potential limitation. A closer examination of previously published studies utilizing this model as it is currently employed reveals that the 'sample size' of these studies is not always as small as it may appear.


Susceptibility of Representative Great Lakes Fish Species to the North Carolina Strain of Spring Viremia of Carp Virus (SVCV).
Spring viremia of carp virus (SVCV) is a notifiable pathogen of the World Organization of Animal Health. Since SVCV was isolated in Lake Ontario in 2007, concern has grown about its spread in the Great Lakes basin and its potential negative impacts on fish species of importance in stock enhancement programs basinwide. The susceptibility of representative fish species from the families Cyprinidae (Fathead Minnow Pimephales promelas, Golden Shiner Notemigonus crysoleucas, Spotfin Shiner Cyprinella spiloptera, and Creek Chub Semotilus atromaculatus), Centrarchidae (Largemouth Bass Micropterus salmoides), Percidae (Walleye Sander vitreus), Salmonidae (Rainbow Trout Oncorhynchus mykiss), and Esocidae (Muskellunge Esox masquinongy) to SVCV was evaluated by experimental infection under laboratory conditions. Morbidity and mortality were recorded, and virus re-isolation, seminested reverse transcription PCR, and histopathological assessments were performed. Using intraperitoneal (i.p.) injection, Fathead Minnows and Golden Shiners were highly susceptible to SVCV (40-70% mortality). All dead or moribund and apparently healthy surviving Fathead Minnows and Golden Shiners were SVCV positive. The SVCV was also detected in challenged but healthy Spotfin Shiners (30%) and Creek Chub (5%). However, noncyprinid species exhibited no morbidity or mortality and were free of SVCV following an observation period of 30 d. In a follow-up experimental challenge, Fathead Minnows and Golden Shiners were SVCV challenged at 103 and 105 PFU/mL by means of waterborne immersion. After immersion, Fathead Minnows and Golden Shiners exhibited characteristic SVCV disease signs, but mortality was less (30% and 10% mortality, respectively) than that in fish with i.p. injections. The SVCV was detected in all mortalities and a subset of healthy Fathead Minnows and Golden Shiners. Necrotic changes were observed in the kidneys, liver, spleen, ovaries, and heart, and other histopathological lesions also occurred. These findings suggest that two of the four cyprinids tested are susceptible to SVCV-induced disease and that all four can act as potential carriers of SVCV in the Laurentian Great Lakes. Received January 11, 2017; accepted July 17, 2017.


Sequence analysis of the cryptic plasmid pMG101 from Rhodopseudomonas palustris and construction of stable cloning vectors.
A 15-kb cryptic plasmid was obtained from a natural isolate of Rhodopseudomonas palustris. The plasmid, designated pMG101, was able to replicate in R. palustris and in closely related strains of Bradyrhizobium japonicum and phototrophic Bradyrhizobium species. However, it was unable to replicate in the purple nonsulfur bacterium Rhodobacter sphaeroides and in Rhizobium species. The replication region of pMG101 was localized to a 3.0-kb SalI-XhoI fragment, and this fragment was stably maintained in R. palustris for over 100 generations in the absence of selection. The complete nucleotide sequence of this fragment revealed two open reading frames (ORFs), ORF1 and ORF2. The deduced amino acid sequence of ORF1 is similar to sequences of Par proteins, which mediate plasmid stability from certain plasmids, while ORF2 was identified as a putative rep gene, coding for an initiator of plasmid replication, based on homology with the Rep proteins of several other plasmids. The function of these sequences was studied by deletion mapping and gene disruptions of ORF1 and ORF2. pMG101-based Escherichia coli-R. palustris shuttle cloning vectors pMG103 and pMG105 were constructed and were stably maintained in R. palustris growing under nonselective conditions. The ability of plasmid pMG101 to replicate in R. palustris and its close phylogenetic relatives should enable broad application of these vectors within this group of alpha-proteobacteria.


Clinical, dermoscopic, and histopathologic evaluation of vitamin C versus PRP, with microneedling in the treatment of mixed melasma: A split-face, comparative study.
Melasma is a common esthetic problem affecting the face with a lot of risk factors being incriminated. Although several treatment options are available, none of them is satisfactory. This split face prospective study aimed to compare the efficacy of microneedling with vitamin C versus with platelet-rich plasma (PRP) in the management of mixed melasma. Ten females with bilateral mixed facial melasma were treated with six sessions of microneedling. After the needling vitamin C was applied on the right side of the face and PRP was applied on the left side. Clinical, dermoscopic, and histological assessment of the used treatments was done 1 month after the last session. The clinical and dermoscopic clearance of melasma was proved significantly on both sides of the face but was more significant with vitamin C (P = 0.005). Reduction of epidermal melanin and dermal melanophages was more observed with vitamin C. Moreover, MART-1 stain revealed a more significant reduction in the epidermal, dermal, and the total MART-1 positive cells with vitamin C (P = 0.044, 0.039, and 0.035, respectively). Microneedling with vitamin C was more efficient in treating mixed melasma than with PRP.


Using a relational database to support nursing research.
Designing, implementing, and maintaining a relational database for a complex, longitudinal clinical research project can be the key to the success of a research project. Related issues of data entry, accuracy, confidentiality, security, data analysis, and evaluation of research activities are among the considerations that must be addressed. Our experience with designing a system that was effective and user-friendly to manage the data collected during a 6-year, National Institutes of Health-funded, nursing research project is highlighted. Nurses and other healthcare providers may find our experience with a relational database helpful and applicable to similar clinical research projects.


The revision of a workload measurement tool to reflect the nursing needs of patients with traumatic brain injury.
Rehabilitation for persons with head injuries is a multifaceted process that requires structure, consistency, and predictability. This article discusses the process followed by nurses on one rehabilitation unit to revise a standard nursing workload measurement tool to reflect the physical, cognitive, behavioral, and psychosocial needs of patients with traumatic brain injury (TBI). Family education and support, a critical component of care, was also addressed. The flexibility of the tool, known as GRASP, allowed the incorporation of neurobehavioral items based on the Rancho Los Amigos cognitive functioning scale and the inclusion of important therapeutic nursing interventions. The revised tool reflects the time required to deliver comprehensive nursing care to TBI patients and, thus, promotes the desired rehabilitation outcomes for patients and their families.


Top-down attentional control in spatially coincident stimuli enhances activity in both task-relevant and task-irrelevant regions of cortex.
Models of selective attention predict that focused attention to spatially contiguous stimuli may result in enhanced activity in areas of cortex specialized for processing task-relevant and task-irrelevant information. We examined this hypothesis by localizing color-sensitive areas (CSA) and word and letter sensitive areas of cortex and then examining modulation of these regions during performance of a modified version of the Stroop task in which target and distractors are spatially coincident. We report that only the incongruent condition with the highest cognitive demand showed increased activity in CSA relative to other conditions, indicating an attentional enhancement in target processing areas. We also found an enhancement of activity in one region sensitive to word/letter processing during the most cognitively demanding incongruent condition indicating greater processing of the distractor dimension. Correlations with performance revealed that top-down modulation during the task was critical for effective filtering of irrelevant information in conflict conditions. These results support predictions made by models of selective attention and suggest an important mechanism of top-down attentional control in spatially contiguous stimuli.


Enzymatic method for determining ketone body ratio in arterial blood.
We have developed a new, sensitive, and rapid method for measuring the ketone body concentration in arterial blood and determining the arterial blood ketone body ratio. The procedure involves the sequential use of the enzymes 3-hydroxybutyrate dehydrogenase (3-HBDH; EC 220.127.116.11) and NADH oxidase, followed by a color-generating reaction with the hydrogen peroxide produced by the oxidase reaction. The amount of oxidized chromogen produced is proportional to the 3-hydroxybutyrate (3-HBA) concentration. The acetoacetate (AcAc) concentration is obtained after complete conversion of the AcAc to 3-HBA, in the presence of 3-HBDH. The total 3-HBA concentration is measured and then subtracted from the total ketone body concentration to give the AcAc concentration. This procedure may be applied to plasma samples and the absorbance change measured with an automated chemistry analyzer. Ketone body concentration may be determined over the range 0 to 400 mumol/L. The analysis takes approximately 12 min and requires only 30 microL of plasma.


An analysis of investors' behavior in Bitcoin market.
As an emerging digital asset, Bitcoin has been traded for more than a decade, reaching an impressively high market capitalization and continuing to expand its volume of trading at a rapid pace. Many countries have legalized or are considering legalizing a trading platform for this asset, and a set of companies worldwide accept it as a medium of exchange. As a result of this expansion, many studies in finance literature have focused on studying the efficiency of this cryptocurrency. In line with this literature, this paper investigates, using the abnormal returns and abnormal trading volumes methodologies, the dynamics of investors' reaction to the arrival of unexpected favorable and unfavorable information regarding the Bitcoin market in the context of the three famous hypotheses: the overreaction, the uncertain information, and the efficient market hypotheses. Overall, we find evidence confirming that the Bitcoin market tends to mature over time. More precisely, over the entire analyzed period, investors behave in accordance with the predictions of the uncertain information hypothesis when positive and negative events occur. However, splitting the timespan into sub-periods provides interesting insights. Remarkably in this respect is the fact that starting with the second sub-period, the response of investors in the Bitcoin market supports, in a moderate manner, the postulate of the efficient market hypothesis when favorable events are addressed. Moreover, our findings reveal that during the pandemic period, the efficiency of Bitcoin has increased, thus turning this stressful period into an advantage for this cryptocurrency. This improved market efficiency is also supported by the abnormal trading volume analysis.


Analysis of dependent survival data applied to lifetimes of amalgam fillings.
When studying lifetimes of amalgam fillings, one is faced with the fact that each patient may contribute multiple survival data. This creates a dependence problem, and some methods for solving it are presented here. Analysis for individual patients with simple approaches for combining data is discussed first. Then a parametric frailty model is applied to analyse variation within and between patients. The results are presented in the form of an empirical Bayes estimate for each patient.


Lateralization in feeding is food type specific and impacts feeding success in wild birds.
Current research suggests that hemispheric lateralization has significant fitness consequences. Foraging, as a basic survival function, is a perfect research model to test the fitness impact of lateralization. However, our understanding of lateralized feeding behavior is based predominantly on laboratory studies, while the evidence from wild animals in natural settings is limited. Here we studied visual lateralization in yellow-footed green pigeons (Treron phoenicoptera) feeding in the wild. We aimed to test whether different types of food objects requiring different searching strategies elicit different eye/hemisphere biases. When feeding on relatively large, uniformly colored food objects (mahua flowers) which can be present or absent in the viewed patch, the majority of pigeons relied mostly on the left eye-right hemisphere. In contrast, when feeding on smaller and more abundant food objects, with color cues signaling its ripeness (sacred figs), right-eye (left-hemisphere) preference prevailed. Our results demonstrate that oppositely directed visual biases previously found in different experimental tasks occur in natural feeding situations in the form of lateralized viewing strategies specific for different types of food. The results suggest that pigeons rely on the hemisphere providing more advantages for the consumption of the particular type of food objects, implying the relevance of brain lateralization as a plastic adaptation to ecological demands. We assessed the success of food discrimination and consumption to examine the link between lateralization and cognitive performance. The use of the preferred eye resulted in better discrimination of food items. Discrimination accuracy and feeding efficiency were significantly higher in lateralized individuals. The results showed that visual lateralization impacted pigeons' feeding success, implicating important fitness benefits associated with lateralization.


A polyclonal antibody to a synthetic peptide derived from the rat follicle-stimulating hormone receptor reveals the recombinant receptor as a 74-kilodalton protein.
We have prepared a polyclonal antibody (AntiF) against a synthetic peptide comprising residues 19-29 of the rat FSH receptor. The specificity of this antibody was documented using human embryonic kidney (293) cells and stable transfectants of 293 cells expressing the recombinant LH/CG and FSH receptors. The data presented show that AntiF inhibits the binding of FSH, but not that of LH/CG, to their cognate receptors. AntiF also recognizes a specific protein(s) representing the FSH receptor in Western blots or by immunoprecipitation of cells transfected with the FSH receptor. This protein(s) is not present in untransfected 293 cells or in 293 cells permanently transfected with the LH/CG receptor. Finally, addition of the 19-29 peptide prevents immunoprecipitation of the FSH receptor by AntiF, whereas an unrelated peptide corresponding to residues 637-647 has no effect. In Western blots of 293 cells transfected with the FSH receptor, AntiF reveals the recombinant receptor as a heterogenous glycoprotein with a molecular mass of 58,000-83,000 daltons (in the absence of thiol-reducing agents) or 69,000-81,000 (in the presence of thiol reducing agents). Pulse-chase experiments with metabolically labeled cells show that the mature FSH receptor is a 74-kilodalton protein derived from 67- and 72-kilodalton precursors.


Structural distortion induced broad emission in vacancy-ordered halide triple perovskites.
Structural distortion in halide perovskites is important to tune the optical properties of the materials. The octahedra formed by metal cations and halide anions in these classes of materials remain symmetric; however, the introduction of asymmetry provides enormous opportunities to improve the photoluminescence emission and excited-state lifetimes for their application in white light emitters. In this work, we have systematically introduced asymmetry in vacancy-ordered halide triple perovskite materials Cs3M2X9 (M = Bi3+, Sb3+; X = Cl-, Br-, I-) by mixing trivalent sites in three different halide compounds. The Raman and FT-far-IR measurements were used to investigate the distortion introduced in these materials. The distortion is shown to (i) enhance self-trapped excitonic emission, which is broad and intense leading to emission in the complete visible region and (ii) improve excited-state lifetimes. This strategy to create distortion and its proven ability to improve light emission will find application in light-emitting diodes.


Temporal shaping of third-harmonic pulses on the Nova laser system.
We demonstrate temporal shaping of 0.35-microm-wavelength pulses produced by a third-harmonic conversion of the output from the Nova Nd:phosphate glass-laser amplifier system for use in inertial confinement fusion experiments. We describe the computer models used to calculate the pulse shape that is required as the input to the amplifier system, the experimental apparatus used to produce these pulses, and the high-power 0.35-microm shaped pulses produced in recent experiments.


Idiopathic pure sudomotor failure: anhidrosis due to deficits in cholinergic transmission.
Acquired idiopathic generalized anhidrosis (AIGA) represents a heterogeneous clinical syndrome including sudomotor neuropathy and failure of the sweat glands. However, most AIGA cases comprise idiopathic pure sudomotor failure (IPSF), a distinct subgroup without sudomotor neuropathy or sweat gland failure. Eight patients with IPSF (mean +/- SD age 20 +/- 5 years) were assessed by thermoregulatory and pilocarpine-induced sweating tests, as well as emotional sweating using sudorometer (4 cases), microneurography of skin sympathetic nerve activity (2 cases), and skin biopsies from the forearm or axilla (3 cases). Clinical features of IPSF comprise early onset; acute or sudden onset; concomitant sharp pain or cholinergic urticaria over the entire body; lack of autonomic dysfunction other than generalized anhidrosis; elevated serum IgE levels; and marked response to steroid. Sudomotor function testing revealed complete absence of thermoregulatory sweating, but well-preserved emotional sweating; pilocarpine did not induce sweating, and microneurography revealed that bursts of skin sympathetic nerve activity were not decreased; and skin biopsy displayed no morphologic abnormalities in sweat glands. The first two findings suggest lesions on the postsynaptic side of the nerve-sweat gland junction. The lesions in IPSF may be in the muscarinic cholinergic receptors of sweat glands. Allergic mechanisms are probably involved in its pathophysiology.


Assessment of the production of antioxidants from winemaking waste solids.
Winemaking waste solids (WS, resulting from red grapes after fermentation and distillation to recover spirits) were subjected to various processing schemes for isolating fractions with antioxidant activity. The liquors entrapped in WS as received were separated by pressing and freeze-dried to yield a fraction with antioxidant activity (measured as DPPH radical scavenging capacity) comparable to those of synthetic antioxidants. A second approach based on the direct processing of raw WS in sulfuric acid medium under fixed operational conditions and further extraction of hydrolysis liquors with ethyl acetate enabled the isolation of a fraction with higher antioxidant ability at an improved yield. The most favorable approach started with a washing stage leading to liquors (which were directly freeze-dried to yield 1.20 g of extract/100 g of oven-dry WS and presented an EC50 of 0.41 g of extract/L) and washed solids, which were dried and subjected to hydrolytic processing (i) with water as a reactive in an autocatalyzed reaction (autohydrolysis) or (ii) with sulfuric acid solutions to give an ethyl acetate-soluble fraction with improved antioxidant properties (EC50 in the range of 0.18-0.40 g/L). Samples from washing liquors and processing of washed solids in aqueous medium were subjected to chromatographic fractionation and analysis to give isolates with remarkable antioxidant activity (with EC50 as low as 0.07 g/L) and to identify their major components.


Design and Control of a Magnetically-Actuated Capsule Robot With Biopsy Function.
Wireless capsule endoscopy has been well used for gastrointestinal (GI) tract diagnosis. However, it can only obtain images and cannot take samples of GI tract tissues. In this study, we designed a magnetically-actuated biopsy capsule (MABC) robot for GI tract diagnosis. The proposed robot can achieve locomotion and biopsy functions under the control of external electromagnetic actuation (EMA) system. Two types of active locomotion can be achieved, plane motion refers to the robot rolling on the surface of the GI tract with a rotating uniform magnetic field. 3D motion refers to the robot moving in 3D space under the control of the EMA system. After reaching the target position, the biopsy needle can be sprung out for sampling and then retracted under a gradient magnetic field. A pill-shaped robot prototype ( ϕ15 mm×32 mm) has been fabricated and tested with phantom experiments. The average motion control error is 0.32 mm in vertical direction, 3.3 mm in horizontal direction, and the maximum sampling error is about 5.0 mm. The average volume of the sampled tissue is about 0.35 mm 3. We designed a MABC robot and proposed a control framework which enables planar and 3D spatial locomotion and biopsy sampling. The untethered MABC robot can be remotely controlled to achieve accurate sampling in multiple directions without internal power sources, paving the way towards precision sampling techniques for GI diseases in clinical procedures.


Cauliflower mosaic virus: a 420 subunit (T = 7), multilayer structure.
The structures of the Cabb-B and CM1841 strains of cauliflower mosaic virus (CaMV) have been solved to about 3 nm resolution from unstained, frozen-hydrated samples that were examined with low-irradiation cryo-electron microscopy and three-dimensional image reconstruction procedures. CaMV is highly susceptible to distortions. Spherical particles, with a maximum diameter of 53.8 nm, are composed of three concentric layers (I-III) of solvent-excluded density that surround a large, solvent-filled cavity (approximately 27 nm dia). The outermost layer (I) contains 72 capsomeric morphological units, with 12 pentavalent pentamers and 60 hexavalent hexamers for a total of 420 subunits (37-42 kDa each) arranged with T = 7 icosahedral symmetry. CaMV is the first example of a T = 7 virus that obeys the rules of stoichiometry proposed for isometric viruses by Caspar and Klug (1962, Cold Spring Harb. Symp. Quant. Biol. 27, 1-24), although the hexameric capsomers exhibit marked departure from the regular sixfold symmetry expected for a structure in which the capsid protein subunits are quasi-equivalently related. The double-stranded DNA genome is distributed in layers II and III along with a portion of the viral protein. The CaMV reconstructions are consistent with the model based on neutron diffraction studies (Kruse et al., 1987, Virology 159, 166-168) and, together, these structural models are discussed in relation to a replication-assembly model (Hull et al., 1987, J. Cell Sci. (Suppl.) 7, 213-229). Remarkable agreement between the reconstructions of CaMV Cabb-B and CM1841 suggests that other strains of CaMV adopt the same basic structure.


Protective effect of saponin on sperm DNA fragmentation of mice treated with cyclophosphamide.
Cyclophosphamide (CP) is a common chemotherapy drug with the testicular damage. The aim of this study was to investigate the effect of saponin (SP) on the toxicity of CP in the male reproductive system. Following an experimental pilot study for determining SP dose, 40 male mice (32 ± 3 g) were divided into five groups (n = 8): control, sham (normal saline 0.2 ml/day), CP (15 mg/kg/week, intraperitoneally), SP (2.5 mg/kg/day, intraperitoneally) and saponin group with cyclophosphamide (SP + CP). After treatment, the left testes were removed for the measurement of malonedialdehyde (MDA) and total antioxidant capacity (TAC) levels, and sperm DNA fragmentation was assessed by SDFA kit. In the CP group, a significant decrease in motility, viability, count, normal morphology and DNA fragmentation of spermatozoa and TAC was observed, while in MDA level, a significant increase was observed compared with the control group (p < 0.05). Attenuated sperm parameters in CP group improved significantly in SP + CP group (p < 0.05). According to the findings of this study, SP was able to alter the reproductive toxicity of CP in NMRI mice and increase the antioxidant capacity of the testis.


[Analysis of Pollution Characteristics, Meteorological Impact, and Forecast Retrospective During the Spring Festival and the Lantern Festival in "2+26" Cities].
The PM2.5concentration characteristics of "2+26" cities in Beijing-Tianjin-Hebei and the surrounding region during the Spring Festival and the Lantern Festival in 2021 were analyzed. The average daily concentration and hourly concentration of PM2.5 in the three days before and after the Spring Festival and the Lantern Festival from 2016 to 2021 and the characteristics of the PM2.5 chemical components during the Spring Festival and the Lantern Festival from 2019 to 2021 were comparatively analyzed. In addition, the impact of meteorological factors on PM2.5 concentration from 2016 to 2021 and the key factors affecting the forecast result of Beijing during the Spring Festival in 2021 were discussed. The results showed that the combination of fireworks and firecrackers and adverse weather conditions resulted in a moderate to heavy pollution process during the Spring Festival in 2021 and a light to moderate pollution process during the Lantern Festival in "2+26" cities. The average ρ(PM2.5) was 111 μg·m-3. The hourly peak ρ(PM2.5) was 156 μg·m-3, which was the lowest in 2016 to 2021. The three days before and after the Lantern Festival in 2021, the average ρ(PM2.5) of "2+26" cities was 85 μg·m-3, and the hourly peak ρ(PM2.5) was 125 μg·m-3. During this period, the hour number of heavy and severe pollution was the lowest in 2016 to 2021. Component observation results showed that the impact of fireworks and firecrackers during the Spring Festival and the Lantern Festival in 2021 was gradually reduced, and the effect of bans and restrictions for fireworks and firecrackers was significant. The regional pollution was dominated by conventional pollution accumulation and nitrate. The concentration of organic matter in some cities during the Spring Festival was relatively high, which may be related to local industrial emissions and the impact of blasting fireworks and firecrackers. The comparative analysis of meteorological factors and PM2.5 concentration showed that the factors such as wind speed and direction, ground pressure, and relative humidity all affected PM2.5 concentration and the duration of the regional pollution process. The influence of oscillation of the boundary layer transport sink on diffusion conditions in the North China Plain, the location and intensity of the small high pressure in North China and the intensity of its high-altitude moving airflow, and the uncertainty of fireworks displays, as well as the low accuracy of weather forecasts caused by frequent weather changes during the alternate period of winter and spring, could affect the accuracy of air quality forecast results in the northern Beijing-Tianjin-Hebei region. In summary, although the impact of fireworks and firecrackers has been reduced in recent years, in order to improve the air quality, it is recommended to further strengthen the control of fireworks and firecrackers, especially outside the Beijing Fifth Ring Road and Beijing's surrounding cities.


Cytochalasin B-induced ATPase activity of actin: dependence on monomer concentration.
We simultaneously measured polymerization and ATPase activity of actin induced by cytochalasin B. It was found that under all conditions tested, ATPase activity was proportional to the concentration of actin which was unpolymerized. In addition, it was found that conditions increasing ATPase activity also increase the velocity of polymerization. From this we conclude that CB-induced ATPase activity is a property of the actin monomers and that the "readiness" of the monomers to hydrolyze ATP is correlated with an increased capacity of the monomers to polymerize.


Biotechnological developments in New Zealand.
Biotechnology has a long history in New Zealand, and there have been some innovations that are unique to the country. This article describes some developments that illustrate this point and gives some examples of current research projects that typify the strengths and interests of the biotechnology community. Examples are taken from the food industry, agriculture, forestry, production of industrial chemicals, and the treatment of hazardous chemicals.


Copper Concentrations in Breast Cancer: A Systematic Review and Meta-Analysis.
Breast cancer is the most common neoplasm, comprising 16% of all women's cancers worldwide. Research of Copper (Cu) concentrations in various body specimens have suggested an association between Cu levels and breast cancer risks. This systematic review and meta-analysis summarize the results of published studies and examine this association. We searched the databases PubMed, Scopus, Web of Science, and Google Scholar and the reference lists of relevant publications. The Standardized Mean Differences (SMDs) between Cu levels in cancer cases and controls and corresponding Confidence Intervals (CIs), as well as I2 statistics, were calculated to examine heterogeneity. Following the specimens used in the original studies, the Cu concentrations were examined in three subgroups: serum or plasma, breast tissue, and scalp hair. We identified 1711 relevant studies published from 1984 to 2017. There was no statistically significant difference between breast cancer cases and controls for Cu levels assayed in any studied specimen; the SMD (95% CI) was -0.01 (-1.06 - 1.03; P = 0.98) for blood or serum, 0.51 (-0.70 - 1.73; P = 0.41) for breast tissue, and -0.88 (-3.42 - 1.65; P = 0.50) for hair samples. However, the heterogeneity between studies was very high (P < 0.001) in all subgroups. We did not find evidence for publication bias (P = 0.91). The results of this meta-analysis do not support an association between Cu levels and breast cancer. However, due to high heterogeneity in the results of original studies, this conclusion needs to be confirmed by well-designed prospective studies.


Comparing immigrant children with native Greek in self-reported-Quality of Life.
Research on an international and national context regarding immigrant children and adolescents' quality of life (QoL) is rather scarce. Few international studies have been conducted investigating the presence of psychopathology and providing evidence of behavioural and psychological problems in immigrant adolescents. Regarding immigrant quality of life, thus far investigation was directed mainly to adult immigrant individuals and not to their children. The aim of the present study was to investigate the quality of life (QoL) of immigrant children and young adolescents who live in the greater Athens area, and to compare them with their native Greek peers living in the same communities and attending the same schools. Sixty three immigrant children, from Albanian and Eastern European origin (mean age 11.9 years) and 489 native children (mean age 11.33 years) were administered a QoL instrument specifically developed for children and adolescents: the Kid-KINDL® Questionnaire for 8-12 years old and the Kiddo-KINDL® Questionnaire for 13-16 years old. The dimensions examined in the KINDL® questionnaire refer to: physical wellbeing, emotional well-being, self- esteem, friends, family life and everyday life (school life). The Greek version is reported to show satisfactory values of validity and reliability. Administration of questionnaires was conducted at school after parent consent. Analysis included student's t-test, chi-square test, and multivariate linear regression analysis, as to investigate the relationship between KINDL® QoL dimensions' scores and nationality status, after controlling for gender and age. The results indicated that self-reported QoL scores of immigrant children were significantly poorer in comparison to native children in the domains of self-esteem and family life, as well as in the total QoL scores. In the rest of the QoL domains, similar scores were reported in both immigrant and their native classmates, that is in the dimensions of physical well-being, emotional well-being, friends and school. Investigating the effect of gender in KINDL® QoL parameters, after controlling for nationality and age, no evidence was found for differences between male and female children. Regarding the effect of age, older compared to younger in age children scored significantly lower in emotional well-being, self-esteem, and friends, school and total QoL. The results provide evidence of QoL deficits in self-esteem and family life in immigrant children. QoL deficits seem to increase in more areas as immigrant children grow older. Dimensions regarding self-perception and family may be interrelated, mutually influencing one another. It may be suggested that immigrant children seem to experience reduced self-esteem and distressful feelings within family interpersonal relations. As they grow older, distress seems to become more evident in emotional and social areas of QoL. Mental health interventions should take into account the multilevel impact of family interpersonal experiences on child's psychosocial development, as to design and deliver appropriate interventions supporting parenting for immigrant groups of individuals. Also, specialized mental health promotion programs need to be provided for adolescent immigrant individuals.


Time-resolved spectroscopic studies of the AppA blue-light receptor BLUF domain from Rhodobacter sphaeroides.
AppA is a blue-light and redox-responding regulator of photosynthesis gene expression in Rhodobacter sphaeroides. Detailed time-resolved fluorescence spectroscopy and subpicosecond transient absorption spectroscopy study of the BLUF domain is presented for wild-type AppA (AppAwt) and a photoinactive Y21F mutant of AppA. The main findings discussed here are that (1) time-resolved laser excitation studies on dark-adapted protein show that AppAwt and Y21F mutant protein exhibits a fluorescence decay with a lifetime of 0.6 ns. Dark-adapted AppAwt but not Y21F also exhibits slower fluorescence decay with a lifetime of 1.7 ns. Analysis of AppAwt that was light-excited to a stable light-adapted form prior to data collection shows monoexponential fluorescence decay with a lifetime of 1.0 ns. This component disappeared after 1 min of data collection after which the original "dark-adapted" values were recovered, demonstrating the presence of a approximately 1 min lifetime intermediate during the return of AppA from light- to dark-adapted form. (2) Transient absorption spectral analysis reveals a very fast rising of transient absorption (<1 ps) for AppAwt. This fast component is missing in the Y21F mutant, which lacks Tyr21, giving rise to a slower transient absorption at 4-6 ps. In the AppAwt transient spectra, most ground states recover within approximately 30 ps, compared to approximately 90-130 ps in the mutant Y21F. We propose that a temporary electron transfer occurs from Tyr21 to the N5 of flavin in AppAwt and is a triggering event for subsequent hydrogen-bond rearrangements. Dynamics of the AppA photocycle is discussed in view of the currently solved crystallographic structure of AppA.


The use of Ziehl-Neelsen stain, enzyme-linked immunosorbent assay and nested polymerase chain reaction in diagnosis of cryptosporidiosis in immuno-competent, -compromised patients.
In the present study, Cryptosporidium parvum was diagnosed in stool by Z-N stain, ELISA and PCR. The detected cases were 5.3%, 8.3% and 9.6% by the previous three methods, respectively. The sensitivity, specificity and accuracy of the different techniques were evaluated. The Z-N stain showed the lowest sensitivity and accuracy in relation to either ELISA or PCR. Moreover, the study revealed that the sensitivity, specificity and accuracy of ELISA detection of Cryptosporidium in relation to detection of DNA in stool by PCR were 84.2%, 96% and 88.8%, respectively. Consequently, PCR showed the best results. From a practical point of view, ELISA is recommended for wide spread use in diagnosis of cryptosporidiosis.


High quality single shot diffraction patterns using ultrashort megaelectron volt electron beams from a radio frequency photoinjector.
Single shot diffraction patterns using a 250-fs-long electron beam have been obtained at the UCLA Pegasus laboratory. High quality images with spatial resolution sufficient to distinguish closely spaced peaks in the Debye-Scherrer ring pattern have been recorded by scattering the 1.6 pC 3.5 MeV electron beam generated in the rf photoinjector off a 100-nm-thick Au foil. Dark current and high emittance particles are removed from the beam before sending it onto the diffraction target using a 1 mm diameter collimating hole. These results open the door to the study of irreversible phase transformations by single shot MeV electron diffraction.


Improving clinical care and elucidating the pathophysiology of Diamond Blackfan anemia: an update from the Diamond Blackfan Anemia Registry.
Diamond Blackfan anemia (DBA) is a heterogeneous genetic disorder characterized by red cell aplasia, congenital anomalies, and a predisposition to cancer. Although incompletely understood, the erythroid failure in DBA appears to result from the accelerated apoptosis of affected erythroid progenitors/precursors. One of what appears to be multiple DBA genes, coding for a ribosomal protein RPS 19, has been cloned. Even within multiplex families individuals may vary dramatically as to the degree of anemia, response to treatment and the presence of congenital anomalies. The Diamond Blackfan Anemia Registry (DBAR), a comprehensive database of pediatric and adult patients with DBA who are enrolled after informed consent, was designed to overcome two significant obstacles encountered in the study of a rare disease; the reporting bias inherent in the literature and the lack of an active patient database. To enroll, patients, their families and their physicians complete a detailed questionnaire. A review of medical records and telephone interviews are performed to complete and clarify the information provided. As of May 1, 2005, 420 patients have been enrolled in the DBAR. Epidemiological, clinical, and laboratory data have been collected and analyzed. The DBAR has provided new information on the clinical presentation, outcome and genetics of DBA as well as a better description of congenital malformations and cancer predisposition. This has resulted both in improved clinical care of patients with DBA as well as providing new insights into the pathophysiology of this complex disorder.


Perceived Congruency in Audiovisual Stimuli Consisting of Gabor Patches and AM and FM Tones.
Experiments that focus on how humans perceive temporal, spatial or synaesthetic congruency in audiovisual sensory information have often employed stimuli consisting of a Gabor patch and an amplitude (AM) or frequency (FM)-modulated sound. Introducing similarity between the static and dynamic features of the Gabor patch and the (carrier) frequency or modulation frequency of the sound is often assumed to be effective enough to induce congruency. However, comparative empirical data on perceived congruency of various stimulus parameters are not readily available, and in particular with respect to sound modulation, it is still not clear which type (AM or FM) induces perceived congruency best in tandem with various patch parameters. In two experiments, we examined Gabor patches of various spatial frequencies with flickering (2, 3 and 4 flickers/s) or drifting (0.5, 1.0 and 1.5 degrees/s) gratings in combinations with AM or FM tones of 2-, 3- and 4-Hz modulation and 500-, 1000- and 2000-Hz carrier frequencies. Perceived congruency ratings were obtained by asking participants to rate stimulus (in)congruency from 1 (incongruent) to 7 (congruent). The data showed that varying the spatial frequency of the Gabor patch and the carrier frequency of the modulated tone had comparatively little impact on perceived congruency. Similar to previous findings, similarity between the temporal frequency of the Gabor patch and the modulated tone effectively promoted perceived congruency. Furthermore, direct comparisons convincingly showed that AM tones in combination with flickering Gabor patches received significantly higher audiovisual congruency ratings compared to FM tones.


The role of stimulus complexity, spectral overlap, and pitch for gap-detection thresholds in young and old listeners.
Thresholds for detecting a gap between two complex tones were determined for young listeners with normal hearing and old listeners with mild age-related hearing loss. The leading tonal marker was always a 20-ms, 250-Hz complex tone with energy at 250, 500, 750, and 1000 Hz. The lagging marker, also tonal, could differ from the leading marker with respect to fundamental frequency (f0), the presence versus absence of energy at f0, and the degree to which it overlapped spectrally with the leading marker. All stimuli were presented with steeper (1 ms) and less steep (4 ms) envelope rise and fall times. F0 differences, decreases in the degree of spectral overlap between the markers, and shallower envelope shape all contributed to increases in gap-detection thresholds. Age differences for gap detection of complex sounds were generally small and constant when gap-detection thresholds were measured on a log scale. When comparing the results for complex sounds to thresholds obtained for pure-tones in a previous study by Heinrich and Schneider [(2006). J. Acoust. Soc. Am. 119, 2316-2326], thresholds increased in an orderly fashion from markers with identical (within-channel) pure tones to different (between-channel) pure tones to complex sounds. This pattern of results was true for listeners of both ages although younger listeners had smaller thresholds overall.