High immunogenicity of a hydrophilic component of the hepatitis B virus preS1 sequence exposed on the surface of three virus-like particle carriers.
The preS1phil, a hydrophilic component of the hepatitis B virus (HBV) preS1 sequence, was exposed on the surface of three widely used virus-like particle (VLP) carriers by (i) insertion into the HI loop of the murine polyomavirus (MPyV) VP1, (ii) N-terminal addition to the hepatitis B surface (HBs) protein, and (iii) insertion into the major immunodominant region (MIR) of three hepatitis B core (HBc) vectors with different structure of their C-termini. Adjuvant-free immunisation of Balb/c mice demonstrated high preS1-specific antibody responses, but strong Th1 cell activation with efficient induction of IgG2a isotype antibodies was observed only in those VLPs, and namely in two of three HBc derivatives, which contained packaged RNA.


Risk and protective factors in the origin of conotruncal defects of heart--a population-based case-control study.
Congenital heart defect (CHD) cases have been evaluated together as a group in some previous epidemiological studies. However, different CHD entities have different etiologies, and the underlying causes are unclear in the vast majority of patients. Thus the aim of this study was to analyze the possible association of different maternal diseases with the risk of four types of conotruncal defects (CTD), that is, truncus arteriosus, d-transposition of the great arteries, tetralogy of Fallot, and double-outlet right ventricle based on autopsy or surgical report diagnosis. Acute and chronic diseases with related drug treatments and peri-conceptual folic acid or multivitamin supplementations were compared in mothers of 598 CTD cases, of 902 matched controls, and 38,151 population controls without any defects, and with 20,896 malformed controls with other isolated non-cardiac defects in the population-based large dataset of the Hungarian Case-Control Surveillance of Congenital Abnormalities. Mothers who had medically recorded influenza and the common cold with secondary complications in the prenatal maternity logbook during the second and/or third gestational months were associated with a higher risk of CTD (OR with 95% CI: 2.22, 1.19-3.88). The common denominator of these maternal diseases may be high fever, which could be prevented by antifever therapies. On the other hand, high doses of medically recorded folic acid in early pregnancy were able to reduce the birth prevalence of CTD (OR with 95% CI: 0.54, 0.39-0.73), and this reduction was significant in transposition of the great arteries (0.46, 0.29-0.71) as well. In conclusion, high fever related maternal diseases may have a role in the origin of CTD, while high doses of folic acid in early pregnancy were able to reduce of CTD, particularly transposition of great vessels.


A comparative analysis of food-niche relationships and trophic guild structure in two assemblages of vertebrate predators differing in species richness: causes, correlations, and consequences.
We conducted a comparative analysis of foodniche relationships and trophic guild structure in two assemblages of vertebrate predators (including hawks, owls, snakes, and carnivores) living in similar habitats of Chile and Spain, which differed in species richness (11 and 25 predator species, respectively), to explore how the structure of predator assemblages reflects an increase (or decrease) in the number of coexisting species. Our results indicate that the Spanish assemblage appears enriched by the symmetrical "addition" of species with disparately large and small values of body weight, diet breadth, and mean prey size, around the same median value found in Chile. Or, alternatively, the Chilean assemblage appears impoverished by the symmetrical "loss" of peripheral species - those at both tails of the frequency distribution of the above trophic estimators. Spanish predators overlap less extensively among themselves, and with smaller variance, than Chilean predators. Consequently, the higher predator richness in Spain is not ascribable to their narrower diet breadths or more extensive diet overlaps as predicted by some theoretical models. Instead, it is associated with a more spread-out use of the available resources. Trophic guilds can be recognized in the two countries, but in Spain guilds are less tightly packed. Some taxomically unrelated species are close diet analogues between Spain and Chile, whereas some closely related species cannot be matched well between countries. Circumstantial evidence suggests that the presence of some trophic guilds and of some specialist predators in either country is based on the high abundance attained by some particular prey types.


Infrared and time-resolved fluorescence spectroscopic studies of the polymorphic phase behavior of phosphatidylethanolamine/diacylglycerol lipid mixtures.
Fourier transform infrared (FTIR) and time-resolved fluorescence spectroscopy have been employed to examine the structural dynamics of lipid fatty acyl chains and lipid/water interfacial region of a binary lipid mixture containing unsaturated phosphatidylethanolamine (PE) and diacylglycerol (DG). Infrared vibrational frequencies of the CH2 symmetric stretching and the C = O stretching bands of the lipids were measured at different lipid compositions and temperatures. For 0% DG, the lamellar gel to lamellar liquid crystalline (L beta-L alpha) and the L alpha to inverted hexagonal (L alpha-HII) phase transitions were observed at approximately 15 degrees and 55 degrees C, respectively. As the DG content increased gradually from 0% to 15%, the L alpha-HII phase transition temperature decreased drastically while the L beta-L alpha phase transition temperature decreased only slightly. At 10% DG, a merge of these two phase transitions was noticed at approximately 10 degrees C. For the composition study at 23 degrees C, the L alpha-HII transition occurred at approximately 6-10% DG as indicated by abrupt increases in both the CH2 and C = O stretching frequencies at those DG contents. Using time-resolved fluorescence spectroscopy, abrupt decreases in both the normalized long time residual and the initial slope of the anisotropy decay function of lipid probes, 1-palmitoyl-2-[[2-[4-(6-phenyl-trans-1,3,5- hexatrienyl)phenyl]ethyl]carbonyl]-3-sn-phosphatidylcholine, in these PE/DG mixtures were observed at the L alpha-HII phase transition. These changes in the anisotropy decay parameters suggested that the rotational dynamics and orientational packing of the lipids were altered at the composition-induced L alpha-HII transition, and agreed with a previous temperature-induced L alpha-HII transition study on pure unsaturated PE (Cheng (1989) Biophys. J. 55, 1025-1031). The fluorescence lifetime of water soluble probes, 8,1-anilinonapthalenes sulfonate acid, in PE/DG mixtures increased abruptly at the L alpha-HII phase transition, suggesting that the conformation and hydration of the lipid/water interfacial region also undergo significant changes at the L alpha-HII transition.


Integrated multiomics approach identifies calcium and integrin-binding protein-2 as a novel gene for pulse wave velocity.
Carotid-femoral pulse wave velocity (PWV) is an important measure of arterial stiffness, which is an independent predictor of cardiovascular morbidity and mortality. In this study, we used an integrated genetic, epigenetic and transcriptomics approach to uncover novel molecular mechanisms contributing to PWV. We measured PWV in 1505 healthy twins of European descendent. A genomewide association analysis was performed using standardized residual of the inverse of PWV. We identified one single-nucleotide polymorphism (rs7164338) in the calcium and integrin-binding protein-2 (CIB2) gene on chromosome 15q25.1 associated with PWV [β = -0.359, standard error (SE) = 0.07, P = 4.8 × 10]. The same variant was also associated with increased CIB2 expression in leucocytes (β = 0.034, SE = 0.008, P = 4.95 × 10) and skin (β = 0.072, SE = 0.01, P = 2.35 × 10) and with hypomethylation of the gene promoter (β = -0.899, SE = 0.098, P = 3.63 × 10). Our data indicate that reduced methylation of the CIB2 promoter in individuals carrying rs7164338 may lead to increased CIB2 expression. Given that CIB2 is thought to regulate intracellular calcium levels, an increase in protein levels may prevent the accumulation of serum calcium and phosphate, ultimately slowing down the process of vascular calcification. This study shows the power of integrating multiple omics to discover novel cardiovascular mechanisms.


Flexibility of telomerase in binding the RNA template and DNA telomeric repeat.
Telomerase synthesizes telomeres at the ends of linear chromosomes by repeated reverse transcription from a short RNA template. Crystal structures of Tribolium castaneum telomerase reverse transcriptase (tcTERT) and cryoelectron microscopy (cryo-EM) structures of human and Tetrahymena telomerase have revealed conserved features in the reverse-transcriptase domain, including a cavity near the DNA 3' end and snug interactions with the RNA template. For the RNA template to translocate, it needs to be unpaired and separated from the DNA product. Here we investigate the potential of the structural cavity to accommodate a looped-out DNA bulge and enable the separation of the RNA/DNA hybrid. Using tcTERT as a model system, we show that a looped-out telomeric repeat in the DNA primer can be accommodated and extended by tcTERT but not by retroviral reverse transcriptase. Mutations that reduce the cavity size reduce the ability of tcTERT to extend the looped-out DNA substrate. In agreement with cryo-EM structures of telomerases, we find that tcTERT requires a minimum of 4 bp between the RNA template and DNA primer for efficient DNA synthesis. We also have determined the ternary-complex structure of tcTERT including a downstream RNA/DNA hybrid at 2.0-Å resolution and shown that a downstream RNA duplex, equivalent to the 5' template-boundary element in telomerase RNA, enhances the efficiency of telomere synthesis by tcTERT. Although TERT has a preformed active site without the open-and-closed conformational changes, it contains cavities to accommodate looped-out RNA and DNA. The flexible RNA-DNA binding likely underlies the processivity of telomeric repeat addition.


Abnormal head nociceptive withdrawal reaction to facial nociceptive stimuli in Parkinson's disease.
Trigemino-cervical-spinal reflexes (TCSRs) are complex brainstem stereotyped nociceptive responses involved in a defensive withdrawal reaction of the head from facial nociceptive stimuli. The present study was undertaken to collect data on possible TCSR abnormalities in idiopathic Parkinson's disease (PD) and investigate any correlation with motor signs and L-DOPA administration. TCSRs were registered from the semispinalis capitis and biceps brachii muscles after electrical stimulation of the supraorbital nerve in 18 patients with PD and 24 controls. The latency (L) and area (A), as well as the sensory (ST), painful (PT) and reflex (RT) thresholds were measured during the 'off' and 'on' state, and possible correlations with the UPDRS III total score, selected subscores (tremor, neck rigidity, upper limb rigidity, akinesia, rising from a chair, posture and posture instability) and duration of illness were investigated. Significant changes between controls and PD patients were found in the L, A, PT and RT of TCSRs. These results were not significantly influenced by L-DOPA treatment. A significant correlation was found between neck rigidity, postural instability scores and duration of illness and the TCSR L and A values in PD patients in the 'off' state. TCSRs abnormalities, combined with dopamine resistance, are consistent with a primary loss of brainstem neurons mediating a complex sensory-motor integration including neck muscle tone and postural control as well as the head withdrawal reaction to the nociceptive stimuli. TCSRs may represent a useful tool for the assessment of brainstem sensory-motor function in PD as well as other movement and degenerative disorders.


Dissociation between the Espinet and perceptual learning effects in flavour aversion conditioning.
In three experiments rats were given pre-exposure to two compound flavours, AX and BX, the two compounds being presented for some subjects on alternate trials (the intermixed schedule) and, for others, in separate blocks of trials (the blocked schedule). After aversion conditioning with A (in Experiments 1 and 2), the inhibitory properties of B were tested using both retardation (Experiment 1) and summation tests (Experiment 2). The results failed to support the proposal [Anim. Learn. Behav. 23 (1995) 361] that B should acquire inhibitory properties in the intermixed condition (the "Espinet effect"). Experiment 3 demonstrated that generalisation to BX after conditioning with AX was attenuated by intermixed pre-exposure (a perceptual learning effect). This pattern of results challenges the hypothesis that inhibitory learning during intermixed pre-exposure to AX and BX can account for both the Espinet and the perceptual learning effects.


Therapeutic Pitfalls in the Transition of Neurologic Patients from Pediatric to Adult Health Care Providers.
Transitions of care from the pediatric to adult setting are fraught with difficulty. For patients whose neurologic problems began in childhood, there is often a lack of organized multidisciplinary care with the desired neurologic expertise in the adult setting. This monograph highlights those difficulties, reviewing disease-specific instances of the problems with transition from pediatric to adult neurologic care. The use of an arbitrary chronologic age cutoff for transition from pediatric to adult expertise in specific disease state may be a disservice in provision of care, and the disease-specific expertise of providers may outweigh the benefit of an age-related provider focus.


Wax lipid secretion and ultrastructural development in the egg-waxing (Gene's) organ in ixodid ticks.
Gene's organ, the egg-waxing organ of ticks, performs an essential function in females by coating the eggs with a waterproofing layer during oviposition, which prevents desiccation of the embryo, ensuring its viability. The organ is a target for control agents and a potential site of virus replication involving trans-oval transmission of arboviruses. The organ is a complex dermal gland, developed to an elaborate degree. The external appendage, the horns, is an evertable balloon-like cuticular sac which manipulates the eggs and coats them in wax. Wax passes through pores in the cuticle from the internal, sub-cuticular lumen. Gene's organ develops in synchrony with oogenesis and oviposition. This paper describes the development of the gland cells and formation of the intra-cuticular lumen and its ultrastructure during engorgement and oviposition in ixodid ticks. The structural basis for wax secretion in Gene's organ is also described.


Neurexins play a crucial role in cerebellar granule cell survival by organizing autocrine machinery for neurotrophins.
Neurexins (NRXNs) are key presynaptic cell adhesion molecules that regulate synapse formation and function via trans-synaptic interaction with postsynaptic ligands. Here, we generate cerebellar granule cell (CGC)-specific Nrxn triple-knockout (TKO) mice for complete deletion of all NRXNs. Unexpectedly, most CGCs die in these mice, and this requirement for NRXNs for cell survival is reproduced in cultured CGCs. The axons of cultured Nrxn TKO CGCs that are not in contact with a postsynaptic structure show defects in the formation of presynaptic protein clusters and in action-potential-induced Ca2+ influxes. These cells also show impaired secretion of depolarization-induced, fluorescence-tagged brain-derived neurotrophic factor (BDNF) from their axons, and the cell-survival defect is rescued by the application of BDNF. These results suggest that CGC survival is maintained by autocrine neurotrophic factors and that NRXNs organize the presynaptic protein clusters and the autocrine neurotrophic-factor secretory machinery independent of contact with postsynaptic ligands.


Identification and DNA sequence of a human apolipoprotein E cDNA clone.
cDNA clones encoding human apolipoprotein E were identified by screening an adult human liver cDNA library with an oligonucleotide probe. The probe was a mixture of synthetic 14-base long DNA oligomers constructed to correspond to all possible codons for apo-E amino acids 218-222. Plasmids from four of the 20 clones selected by this screening procedure were digested with PstI and all had five internal PstI sites with a total length of the cDNA insert of approximately 900 base pairs. DNA sequence analysis of one of these clones, designated pE-301, revealed that it corresponded to apo-E amino acids 81-299, and contained a standard termination codon, polyadenylation signal, and poly A tail. The DNA sequence examined included the known apo-E polymorphic sites at amino acids 112, 145, and 158, and the mutant apo-E phenotypes can all be explained on the basis of a single base substitution in the first position of each of these codons. This work supports the hypothesis that the apo-E polymorphism is due to mutations in the region of DNA coding for the apo-E structural gene.


Platelets, coronary heart disease, and stress.
Coronary heart disease is the leading cause of death in Western society, and its development is associated with chronic stress and other psychosocial factors. Atherosclerosis, the disorder underlying this disease, is an inflammatory process in which leukocytes interact with structurally intact but dysfunctional endothelium of the arteries. Platelets play a key role in this process by binding to leukocytes and promoting their recruitment to the endothelium. Platelet-leukocyte interactions also stimulate the release of pro-inflammatory and pro-thrombotic factors which promote atherosclerosis. Elevated circulating levels of platelet-leukocyte aggregates have been reported in cardiac patients and in individuals of low socioeconomic status, a factor associated with chronic psychological stress. Increased platelet activation has also been observed in individuals prone to depression or hostility, and in people subject to high levels of work stress. Acute psychological stress increases circulating platelet-leukocyte aggregates in healthy individuals and this effect is prolonged in cardiac patients. Platelet activation may be a mechanism linking psychosocial stress with increased coronary risk, and may also play a role in the emotional triggering of acute coronary syndromes in patients with advanced coronary disease.


Subchondroplasty in the treatment of bone Marrow lesion in early Knee Osteoarthritis: A systematic review of clinical and radiological outcomes.
Knee osteoarthritis (KOA) is increasingly prevalent in North American society. The significant societal burden it represents makes it essential to promote and target new treatments in earlier phases of the disease. Among others, subchondroplasty is a newly documented technique using calcium phosphate injection targeting the osteochondral lesions preceding KOA, also known as Bone Marrow Lesions (BMLs). This article aimed to review the existing literature on clinical and radiological outcomes of subchondroplasty in the treatment of BMLs in KOA. A systematic review was performed using PubMed, Embase, Medline and Cochrane Database of Systematic Reviews. Studies on calcium phosphate injections into BMLs for KOA and its clinical and radiological outcomes were screened and reviewed by independent evaluators. After screening, ten articles were included, totaling 540 patients. Follow-up ranged from 6 months to 7 years. Overall, the procedure showed significant functional and quality of life improvement, as well as pain relief, as shown by Patients-Reported Outcomes Measures (PROMs). There were very few complications reported, the most important being leakage of calcium phosphate outside the targeted site. Conversion rate to total knee arthroplasty (TKA) ranged from 14 % to 30 % at 2 years post-procedure. Long term radiological outcomes have been poorly documented. Subchondroplasty is a promising avenue for the treatment of KOA. However, quality evidence is still required before any real conclusions and practical management guidelines can be drawn. Prospective, randomized studies with a control group and a rigorous assessment of long-term clinical and radiological outcomes are recommended.


Stress Is Associated With Neurocognitive Function in Hispanic/Latino Adults: Results From HCHS/SOL Socio-Cultural Ancillary Study.
The purpose of this study was to evaluate the hypothesis that chronic and acculturative stress would be negatively associated with neurocognitive function among middle aged to older Hispanics/Latinos. Our analytic sample consisted of 3,265 participants (mean age = 56.7 (±0.24)) from the Hispanic Community Health Study/Study of Latinos who participated in its Sociocultural Ancillary Study. During the baseline phase of this project, participants were assessed on multiple domains of neurocognitive function, and completed self-report measures of chronic and acculturative stress. Each standard deviation increase in chronic stress was associated with lower performance in a verbal learning task (B = -.17, 95% CI [-.32, -.01]); this association was no longer significant after adjusting for mental and physical health symptoms, including depression and anxiety symptoms, and cardiovascular health. A standard deviation increase in acculturative stress was associated with poorer performance in all cognitive measures (Bs range = -.13 to -1.03). Associations of acculturation stress with psychomotor speed, verbal learning, and word fluency remained significant after adjusting for mental and physical health symptoms. Our results suggest that mental and physical health may help explain some cross-sectional associations between stress and cognition and highlight the need to examine culture-specific psychosocial stressors to better understand the context of psychosocial risk factors for neurocognitive performance.


Probing Chromosome Dynamics in Bacillus subtilis.
Research over the last two decades has revealed that bacterial genomes are, in fact, highly organized. The goal of future research is to understand the molecular mechanisms underlying bacterial chromosome architecture and dynamics during the cell cycle. Here we discuss techniques that can be used with live cells to analyze chromosome structure and segregation in the gram-positive model organism Bacillus subtilis.


Are chemokines the third major system in the brain?
Chemokines are a family of small proteins involved in cellular migration and intercellular communication. Although the chemokines and their receptors are located throughout the brain, they are not distributed uniformly. Among the chemokines and their receptors that are arrayed disproportionately in glia and neurons are monocyte chemotactic protein-1/CC chemokine ligand 2 (CCL2), stromal cell-derived factor-1/CXC chemokine ligand 12 (CXCL12), fractalkine/CX3C chemokine ligand 1, interferon-gamma-inducible-protein-10/CXCL10, macrophage inflammatory protein-1alpha/CCL3, and regulated on activation, normal T cell expressed and secreted/CCL5. In the brain, they are found in the hypothalamus, nucleus accumbens, limbic system, hippocampus, thalamus, cortex, and cerebellum. The uneven distribution suggests that there may be functional roles for the chemokine "system," comprised of chemokine ligands and their receptors. In addition to anatomical, immunohistochemical, and in vitro studies establishing the expression of the chemokine ligands and receptors, there is an increasing body of research that suggests that the chemokine system plays a crucial role in brain development and function. Our data indicate that the chemokine system can alter the actions of neuronally active pharmacological agents including the opioids and cannabinoids. Combined with evidence that the chemokine system in the brain interacts with neurotransmitter systems, we propose the following hypothesis: The endogenous chemokine system in the brain acts in concert with the neurotransmitter and neuropeptide systems to govern brain function. The chemokine system can thus be thought of as the third major transmitter system in the brain.


Evolution, selection and cognition: from "learning" to parameter setting in biology and in the study of language.
Most biologists and some cognitive scientists have independently reached the conclusion that there is no such thing as learning in the traditional "instructive" sense. This is, admittedly, a somewhat extreme thesis, but I defend it here in the light of data and theories jointly extracted from biology, especially from evolutionary theory and immunology, and from modern generative grammar. I also point out that the general demise of learning is uncontroversial in the biological sciences, while a similar consensus has not yet been reached in psychology and in linguistics at large. Since many arguments presently offered in defense of learning and in defense of "general intelligence" are often based on a distorted picture of human biological evolution, I devote some sections of this paper to a critique of "adaptationism," providing also a sketch of a better evolutionary theory (one based on "exaptation"). Moreover, since certain standard arguments presented today as "knock-down" in psychology, in linguistics and in artificial intelligence are a perfect replica of those once voiced by biologists in favor of instruction and against selection, I capitalize on these errors of the past to draw some lessons for the present and for the future.


The transmembrane domain of the amyloid precursor protein is required for antiamyloidogenic processing by α-secretase ADAM10.
Neurotoxic amyloid β-peptides are thought to be a causative agent of Alzheimer's disease in humans. The production of amyloid β-peptides from amyloid precursor protein (APP) could be diminished by enhancing α-processing; however, the physical interactions between APP and α-secretases are not well understood. In this study, we employed super-resolution light microscopy to examine in cell-free plasma membranes the abundance and association of APP and α-secretases ADAM10 (a disintegrin and metalloproteinase) and ADAM17. We found that both secretase molecules localize similarly closely to APP (within ≤50 nm). However, when cross-linking APP with antibodies directed against the GFP tag of APP, in confocal microscopy, we observed that only ADAM10 coaggregated with APP. Furthermore, we mapped the involved protein domain by using APP variants with an exchanged transmembrane segment or lacking cytoplasmic/extracellular domains. We identified that the transmembrane domain of APP is required for association with α-secretases and, as analyzed by Western blot, for α-processing. We propose that the transmembrane domain of APP interacts either directly or indirectly with ADAM10, but not with ADAM17, explaining the dominant role of ADAM10 in α-processing of APP. Further understanding of this interaction may facilitate the development of a therapeutic strategy based on promoting APP cleavage by α-secretases.


Luminescent metal organic frameworks with recognition sites for detection of hypochlorite through energy transfer.
A luminescent metal organic framework (LMOF) of type UiO-66-NH2 was chosen for specific and sensitive detection of trace levels of hypochlorite. Hypochlorite causes the quenching of the blue fluorescence of nano-UiO-66-NH2 (with excitation/emission maxima at 325/430 nm), and this finding forms the basis for a fluorometric assay for hypochlorite. The method overcomes disadvantages of conventional redox-probes which are interfered by oxidants with oxidation capability stronger than that of hypochlorite. Compared with other fluorescent probes for sensing hypochlorite, UiO-66-NH2 has a comparable detection limit of 0.3 μmol L-1 and a broad linearity relationship in the range of 1-8 μmol L-1. The probe was successfully applied to the detection of hypochlorite in complex water samples and living Hela cells. Graphical abstract Schematic representation of hypochlorite induced quenching of the blue fluorescence of nano-UiO-66-NH2 (with excitation/emission maxima at 325/430 nm) through energy transfer. It overcomes disadvantages of conventional redox-probes which are interfered by oxidants with oxidation capability stronger than that of hypochlorite.


Peculiarities of the effect of hormones and transmitters during aging: modulation of changes in dopaminergic action.
Loss of motor control is an important functional manifestation of aging. This dysfunction appears to be at least partially due to loss of striatal dopamine receptors. We have examined the possible mechanisms by which such a deterioration occurs as well as possible manipulations to delay and/or reverse these impairments.


Calcium binding to intestinal goblet cell mucin.
1. Binding of Ca-2+ to goblet cell mucin of rat small intestine was studied using equilibrium dialysis against 0.01 M Tris/HCl buffer (pH 7.4) and tracer amounts of 45-CaCl2. Binding was found to reach saturation at a Ca free -2+ concentration of 0.1--1.0 mM, to be independent of temperature (4-37 degrees C), and to increase with increasing pH (5.0-8.7). At low concentrations of Ca free -2+ (smaller than 0.03 mM) the binding curve was sigmoidal, suggesting positive cooperativity of binding sites and a possible change in the tertiary structure of the mucin. Binding was markedly reduced, and sigmoidicity abolished, by removal of sialic acid from the mucin, or by adding 0.14 M NaCl to the dialysis medium. This latter finding suggests that, in vivo, other cations would compete for Ca-2+ binding ligands. 2. Under conditions mimicking those used for binding studies, CaCl2 (10- minus 5 M) was found to cause a small increase (0.03 units) in the absorbance of mucin solutions, especially in the ultraviolet region, possibly indicating increased light scattering. No change in the solubility of the mucin was observed after the addition of CaCl2 (10- minus 6-10- minus 4 M). A significant decrease in viscosity of the mucin was noted, however, with the addition of CaCl2 (10- minus 6-10- minus 2 M). Together with the binding data, these findings suggested that during binding, Ca-2+ combines with negative charges on goblet cell mucin (especially those of sialic acid carboxyl groups) and induces contraction or folding of the macromolecule which promotes cooperative cation binding. No evidence was obtained to suggest that CaCl2 caused precipitation, polymerization or gelation of the mucin in 0.01 M Tris/HCl.?


No filters, no fridges: a method for preservation of water samples for eDNA analysis.
Advancements in the detection of environmental DNA (eDNA) for detecting species of interest will likely allow for expanded use of these techniques in the field. One obstacle that continues to hinder applications in the field is the requirement of a cold chain of storage for water samples containing eDNA. While eDNA has been successfully preserved using Longmire's lysis buffer applied to filters, it has yet to be tried with freshwater samples collected for eDNA detection of an invasive species. We tested the utility of Longmire's solution (100 mM Tris, 100 mM EDTA, 10 mM NaCl, 0.5 % SDS, 0.2 % sodium azide) as an additive to freshwater samples for preservation of eDNA. Environmental DNA was effectively preserved in 15 mL water samples with Longmire's solution added; eDNA positive detection was comparable to freezing the samples at -80 °C and occurred out to 56 days at the highest concentration (5 mL Longmire's solution: 15 mL sample water). Medium and low concentrations of Longmire's solution added to 15 mL of sample water generally preserved eDNA out to 56 days but not as well as did freezing or application of the highest concentration of Longmire's lysis buffer. Treatment and degradation time had a significant effect on average DNA concentration of samples, although not the interaction of treatment and time. Perfect detection occurred out to 56 days with the high Longmire's treatment group but DNA concentration was significantly lower at this time point compared to 28 days. We conclude that Longmire's lysis buffer is a viable alternative to cold chain storage that can simplify the collection of eDNA by eliminating the need for filtering and allow more time for sample collection when added at our highest concentration (1 part Longmire's:3 parts water sample), which could translate to an increase in the chances of detecting a rare or elusive species.


Acute cytomegalovirus infection and transient carotid intimal-medial thickening in a young, otherwise healthy woman.
Carotid intimal-medial thickening was observed in a 23-year-old woman with acute cytomegalovirus (CMV) infection. The thickening disappeared after her recovery from the infection. As endothelial cells are common targets of CMV, this thickening suggests that CMV infection causes vascular lesions, even in otherwise healthy individuals.


Analysis of Circulating Tumor DNA to Predict Neoadjuvant Therapy Effectiveness and Breast Cancer Recurrence.
Real-time detection and intervention can be used as potential measures to markedly decrease breast cancer mortality. Assessment of circulating tumor DNA (ctDNA) may offer great benefits for the management of breast cancer over time. However, the use of ctDNA to predict the effectiveness of neoadjuvant treatment and recurrence of breast cancer has rarely been studied. We prospectively recruited 31 breast cancer patients with 4 subtypes. Three time points were set in this study, including before any therapy (C1), during surgery (T), and six months after surgery (C2). We collected peripheral blood samples from all 31 patients at C1, tumor tissue from all 31 patients at T, and peripheral blood samples from 25 patients at C2. Targeted 727-gene panel sequencing was performed on ctDNA from all blood samples and tissue DNA from all tissue samples. Somatic mutations were detected and analyzed using a reference standard pipeline. Statistical analysis was performed to identify possible associations between ctDNA profiles and clinical outcomes. In total, we detected 159, 271, and 70 somatic mutations in 30 C1 samples, 31 T samples, and 12 C2 samples, respectively. We identified specific genes, such as PIK3CA, TP53, and KMT2C, which were highly mutated in the tissue samples. Furthermore, mutated KMT2C observed in ctDNA of the C2 samples may be an indicator of breast cancer recurrence. Our study highlights the potential of ctDNA analysis at different timepoints for assessing tumor progression and treatment effectiveness, as well as prediction of breast cancer recurrence.


Psychometric properties of the Insomnia Severity Index for people with chronic obstructive pulmonary disease.
Insomnia is one of the most commonly reported symptoms among people with chronic obstructive pulmonary disease (COPD). Prior research evaluated the psychometric properties of the Insomnia Severity Index (ISI) with various populations, but no studies have examined the measurement properties of the instrument in the COPD population. This study aimed to determine the reliability and validity of the ISI for the COPD population. This study included 96 people with COPD and insomnia. As psychometric properties, the ISI's internal consistency, factor structure, and criterion validity were examined with this sample. Exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) were used to evaluate construct validity. Correlations between scores for the ISI and for measures of depression, anxiety, fatigue, and dyspnea were examined to determine criterion validity. The Cronbach's alpha value for the ISI was 0.79, indicating good internal consistency. In the EFA, a single ISI factor with an eigenvalue of 3.19 accounted for 45.6% of the variance. CFA indicated adequate construct validity, and interference of sleep problems with daytime functioning and level of distress caused by sleep difficulties showed the highest factor loadings (both 0.78). Criterion validity was supported by significant, weak to moderate correlations between scores for the ISI and for measures of depression, anxiety, fatigue, and dyspnea. The results provide evidence that the ISI has good reliability and validity for measuring insomnia severity in the COPD population.


Small bowel tumors: A digestive endoscopy society of Taiwan (DEST) multicenter enteroscopy-based epidemiologic study.
Small bowel (SB) accounts for the majority of gastrointestinal tract but its tumors are rare and always overlooked. In this study, we aimed to evaluate the epidemiology of SB tumors. This multicenter retrospective study utilized endoscopy database from 2006/11 to 2016/07. Baseline demographic characteristics, clinical, radiologic and endoscopic findings were collected. Totally 103 (34 benign, 69 malignant lesions) patients with SB tumors in 1070 enteroscopic examinations were enrolled. There were male preponderance (56.3% males, 43.7% females), both in benign (52.9%, 49.1%) and malignant (58.0%, 42.0%) lesions, except for subtype gastrointestinal stromal tumors (GISTs) (31.6%, 68.4%). The age (mean ± SD) at diagnosis in malignant SB tumors (62.2 ± 15.6) was older than those with benign tumors (50.7 ± 21.4) (p < 0.01). Bleeding (43.7%), abdominal pain (40.8%) and ileus (10.7%) were the most common clinical presentations. Hamartoma (32.4%) and adenoma (14.7%) were the most common benign histology. Four major malignant histological subtypes were lymphomas (29.0%), GISTs (27.5%), adenocarcinomas (26.1%) and metastatic cancers (14.5%). SB adenocarcinoma patients (>60-year-old, 77.8%) were older than lymphomas (60%) and GISTs (50%). Proximally location rates of lymphomas, GISTs, adenocarcinomas were 25.0% (5/20), 84.2% (16/19), and 88.9% (16/18), respectively. This endoscopy-based study revealed the most common histology of benign SB tumors were hamartoma and adenoma, and malignant ones were lymphomas, GISTs, adenocarcinomas and metastatic cancers. Most of them were male gender, except for GISTs, and with proximal location, except for lymphomas. Further large-scale investigation efforts are warranted to elucidate the epidemiology of SB tumors.


Phenotypic diversity and selection in biofortified cassava germplasm for yield and quality root traits.
Increasing carotenoid content and improving other root quality traits has been the focus of cassava biofortification. This study aimed to (i) evaluate the genetic variability for total carotenoid content (TCC), as well as for root yield and root quality attributes; (ii) estimate potentially useful correlations for selection; and (iii) select parents for breeding and estimate the genetic gain. Data from 2011 to 2020 of 265 cassava genotypes with cream and yellow roots were analyzed for dry matter content (DMC), shoot yield, fresh root yield (FRY), dry root yield (DRY), harvest index, average number of roots per plant, starch content, root pulp color, cyanogenic compounds, and TCC. The best linear unbiased predictions showed great phenotypic variation for all traits. Six distinct groups were formed for productive characteristics of root quality, mainly TCC, DMC and FRY. Only TCC showed high broad-sense heritability ( h 2 = 0.72), while the other traits had low to medium magnitude (0.21 ≤ h 2 ≤ 0.60). TCC was strongly correlated with pulp color (r = 0.70), but null significance for DMC. The network analysis identified a clear separation between the agronomic and quality attributes of cassava roots. The selection of the 30 genotypes for recombination in the breeding program has the potential to raise TCC by 27.05% and reduce the cyanogenic compounds content by 23.03%, in addition to increasing FRY and DRY by 22.72% and 22.95%, respectively. This is the first consolidated study on the potential of germplasm for the development biofortified cassava cultivars in Brazil.


Spectroscopic methods for the physical characterization and formulation of nonviral gene delivery systems.
Currently, with the exception of naked DNA formulations, most pharmaceutical preparations of plasmid DNA employ some type of polycationic delivery vector such as synthetic cationic polymers and lipids to enhance delivery. A number of biophysical techniques are readily available for the structural characterization of plasmid DNA within synthetic gene delivery complexes. Here we present applications of ultraviolet (UV) absorption, circular dichroism (CD), infrared (IR), and fluorescence spectroscopies as well as dynamic light scattering to the structural analysis of the oligonucleotide component of nonviral gene delivery vectors. We also illustrate this approach for the investigation of the formulation of lipoplex and polyplex-based gene delivery systems. To summarize such data, we show how the macromolecular complexes can be represented as vectors in a highly dimensional space in which the components of the vector consist of normalized values of experimental parameters measured as a function of different solution conditions such as pH and ionic strength.


Histiocytic and t-cell rich b-cell lymphoma (TCRBCL) of the stomach.
Although stomach is a frequent site of extranodal lymphomas, histiocyte-rich TCRBCL has not yet been described there. Even histology of repeated gastrobiopsies of this uncommon, diffuse, large B-cell lymphoma may be inconclusive and partial gastrectomy cannot be avoided. It is only immunohistology (CD20, CD43, CD68) of the paraffin blocks from the resection specimen that can lead to the final diagnosis of intermediate grade malignant lymphoma.


Nanomedicine for implants: a review of studies and necessary experimental tools.
The response of host organisms (including at the protein and cellular level) to nanomaterials is different than that observed to conventional materials. Nanomaterials are those materials which possess constituents less than 100 nm in at least one direction. This review will first introduce the use of nanomaterials in a variety of implant applications highlighting their promise towards regenerating tissues. Such reviewed studies will emphasize interactions of nanomaterials with various proteins and subsequently cells. Moreover, such advances in the use of nanomaterials as novel implants have been largely, to date, determined by conventional methods. However, the novel structure–property relationships unique for nanosized materials reside at the nanoscale. That is, the novelty of a nanomaterial can only be fully appreciated by characterizing their interactions with biological systems (such as proteins) with nanoscale resolution analytical tools. This characterization of nanomaterials at the nanoscale is critical to understanding and, hence, further promoting increased tissue growth on nanomaterials. For this reason, while more tools are needed for this emerging field, this review will also cover currently available surface characterization techniques that emphasize nanoscale resolution pertinent for characterizing biological interactions with nanomaterials, including attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectroscopy (SIMS), colorimetric biological assays, circular dichroism (CD), and atomic force microscopy (AFM). Only through the coordination of nanoscale analytical tools with studies that highlight mechanisms of increased tissue growth on nanomaterials will we be able to design better implant materials.


Discovery of a structurally novel, potent, and once-weekly free fatty acid receptor 1 agonist for the treatment of diabetes.
Type 2 diabetes mellitus (T2DM) is a lifelong disease that requires long-term medication to control glucose levels, and thereby long-acting drug has been clinically needed for improving medical adherence. The free fatty acid receptor 1 (FFA1) was considered as a promising target for several diseases, such as T2DM, pain and fatty liver. However, no once-weekly FFA1 agonist has been reported until now. Herein, we report the successful discovery of ZLY50, the first once-weekly FFA1 agonist with a completely new chemotype, highly agonistic activity and selectivity on FFA1. Moreover, ZLY50 has enough brain exposure to activate FFA1 in brain, and it is the first orally available FFA1 agonist with analgesic activity. Notably, the long-term anti-diabetic and anti-fatty liver effects of ZLY50 (once-weekly) were better than those of HWL-088 (once-daily), a highly potent FFA1 agonist with far stronger glucose-lowering effect than Phase 3 clinical candidate TAK-875. Further mechanism studies suggested that ZLY50 alleviates fatty liver by regulating the expressions of genes related to lipid metabolism, mitochondrial function, and oxidative stress in liver.


Simple GATCA repeats characterize the X chromosomal heterochromatin of Microtus agrestis, European field vole (Rodentia, Cricetidae).
The sex chromosomes of Microtus agrestis are extremely large due to the accumulation of constitutive heterochromatin. We have identified two prominent satellite bands of 2.0 and 2.8 kb in length after HaeIII and HinfI restriction enzyme digestion of genomic DNA, respectively. These satellites are located on the heterochromatic long arm of the X chromosome as shown using Microtus x mouse somatic cell hybrids. By in-gel hybridization with oligonucleotide probes, the organization of the two satellites was studied: among the many copies of the simple tandem tetranucleotide repeat GATA are interspersed rare single GACA tetramers. One of the satellites also harbours related GGAT simple tandem repeats. In situ hybridizations with plasmid-carried or oligonucleotide GATCA probes show clustered silver grains on the long and short arm of the X chromosome. Interspersion of differently organized (GATA)n elements is also demonstrable in the autosomal complement and on the Y chromosome. These results are discussed in the context of the evolution of vertebrate sex chromosomes in relation to heterochromatin and simple repetitive DNA sequences.


Potential for ambiguity in the interpretation of immunochemical studies on multienzymic proteins. The case of uridylate synthase.
Rabbit antibodies directed against homogeneous uridylate synthase multienzyme from mouse Ehrlich ascites carcinoma precipitate both the orotidine-5'-monophosphate decarboxylase (EC 188.8.131.52) and orotate phosphoribosyltransferase (EC 220.127.116.11) activities of mouse and human erythrocyte uridylate synthase. When the partially purified human enzyme is used as antigen the two activities coprecipitate with the same apparent titer; however, when the mouse carcinoma protein was studied under the same conditions the decarboxylase activity immunoprecipitated with significantly higher avidity than did the transferase activity. Since the mouse multienzyme has been shown to be a single polypeptide that contains both activities (McClard, R.W., Black, M.J., Livingstone, L.R. and Jones, M.E. (1980) Biochemistry 19, 4699-4706), these results were, at face value, surprising. However, when the mouse orotate phosphoribosyltransferase activity (which is largely lost upon dilution into the immunoassay medium) was stabilized with 5-phosphoribosyl 1-pyrophosphate, both enzyme activities displayed the same apparent antibody titer. The immunochemical studies indicate that the antibodies, as a population, preferentially bind to a form or forms of the enzyme which contain(s) denatured transferase domains. A calculation based on a simple model yields a value of approximately 100 for the relative selectivity of the antibody for the denatured form of uridylate synthase. These results illustrate an ambiguity that is inherent in the interpretation of immunochemical studies on such multienzymic proteins; that is, it is possible to conclude incorrectly that two enzyme activities are not functionally associated if one of the catalytic domains is particularly unstable and thereby displays greater immunoreactivity for the specific antiserum.


Human eosinophil major basic protein augments bronchoconstriction induced by intravenous agonists in guinea pigs.
The direct effect of intratracheal (IT) administration of human major basic protein (MBP) on pulmonary inspiratory pressure (PIP), and the effect on agonist-induced change in PIP, were determined in anesthetized, ventilated guinea pigs. 500 micrograms MBP increased PIP from 24.1 +/- 4.3 to 49.8 +/- 7.4 cmH2O (p < 0.002, n = 10). Maximum PIP was achieved within 30 min after 500 micrograms MBP. The direct PIP response to 250 micrograms MBP was not different from vehicle. The PIP responses to intravenous (IV) acetylcholine (Ach) and 5-hydroxytryptamine (5-HT) were measured before and after administration of 250 micrograms MBP (n = 12). MBP caused a modest, but significant potentiation of the increase in PIP induced by 1, 3 and 10 micrograms/kg Ach (24, 32 and 28%, respectively, p < 0.02) and to 1 microgram/kg 5-HT (43% p < 0.02). We conclude that MBP at a dose that does not directly affect inspiratory pressure is capable of augmenting the PIP response to IV Ach and 5-HT in vivo.


Characterization of a phycoerythrin without alpha-subunits from a unicellular red alga.
We describe here the spectral and biochemical properties of a novel biliprotein belonging to the phycoerythrin family, purified from the phycobilisome of a unicellular red alga, Rhodella reticulata strain R6. This biliprotein is assembled from a unique beta-type subunit, chloroplast-encoded, whose hexameric or dodecameric aggregates are stabilized by unusually large linkers (87 and 60 kDa) encoded by the nuclear genome. Although each beta-type subunit bears two phycoerythrobilins and one phycocyanobilin per chain, the linker polypeptides are non-chromophorylated. The apoprotein of the beta-subunit of the R. reticulata R6 phycoerythrin is specified by a monocistronic rpeB chloroplast gene that is split into three exons. We discuss the relationships between R6 beta-phycoerythrin and the previously published polypeptide sequences, the structural consequences due to the absence of an alpha-subunit, and its evolutionary implications.


Reproductive strategies of six herbaceous perennial species in relation to a successional sequence.
Dry weight allocation to vegetative and reproductive structures in six perennial herbs was studied at three successional sites (quarry, grassland and scrub) on the magnesian limestone in County Durham, England, during the summer of 1978. Two species showed little difference between sites but the other four all displayed the same pattern. Population reproductive effort was highest in the quarry and lowest in the grassland. Reproductive effort of flowering individuals however was the same at all three sites, so that between-site differences in population reproductive effort were entirely a consequence of variation in the proportion of plants which flowered. Indirect evidence suggests that initiation of flowering may be triggered by the attainment of a critical leaf weight. It is suggested that the between-site differences can be explained in terms of the levels of competition at each site and that reproductive effort does not necessarily decline uniformly during succession.


A novel test for annexin A5 M2 haplotyping in in vitro fertilization patients and preimplantation embryos.
To develop a test for evaluating the annexin A5 M2 haplotype in in vitro fertilization patients and preimplantation embryos. Test performance was measured by comparing Sanger sequencing of parental blood DNA and quantitative real-time polymerase chain reaction (qPCR) of saliva DNA, 3 fibroblast cell line 7-cell aliquots and their corresponding purified DNA, 123 trophectoderm biopsy samples, and DNA isolated from 1 embryonic stem cell line along with the Mendelian inheritance expectations, embryo Sanger sequencing, and single-nucleotide polymorphism (SNP) microarray-based linkage analysis. Preimplantation genetic testing laboratory research on IVF patient and embryo DNA. An assay was developed for the detection of the M2 haplotype on saliva samples of 6 in vitro fertilization patients. In addition, 13 patients who underwent preimplantation genetic testing with data on parental and embryo biopsy DNA available for research use were evaluated. None. The concordance rates between Sanger sequencing, SNP array-based linkage analysis, and Mendelian inheritance expectations with qPCR. The concordance rate between Sanger sequencing and qPCR was 100% on parental blood DNA and saliva DNA. The sample concordance rate between all replicates of 7-cell aliquots was 100%. The sample concordance rate between 3 cell lines used to prepare 7-cell aliquots and purified genomic DNA was 100%. The concordance rate between qPCR and Sanger sequencing results from a single trophectoderm biopsy and isolated embryonic stem cell line was 100%. The concordance rate of trophectoderm biopsy qPCR results and expectations from Mendelian inheritance rules was 97%; however, when SNP array-based linkage analysis was included, the concordance rate reached 100%. This study resulted in the development of a convenient saliva collection method and qPCR-based genotyping method to screen for the M2 haplotype. In addition, a novel method for testing preimplantation embryos has been established, providing an alternative to the use of low molecular weight heparin, through selection of embryos without the M2 haplotype.


[Radiation therapy of carcinomas of the oral tongue and floor of mouth (experiences with 266 cases from 1950 to 1975) (author's transl)].
We report 266 patients, who were treated in the Universitätsklinik für Radiotherapie und Nuklearmedizin Zürich between 1950 and 1976. The relationship of the radiation dose to the tumor control and complication rate in various stages is analysed. In early stages, local control is achieved by a dose about 2200 ret. This dose is applicable without an undue complication rate by interstitial radiation with or without combination with percutaneous irradiation. The cure rate in this early stage is about 70 to 90% by irradiation. The cure rate in the stage T2N0 is about 50%, in T3N0 about 20% and in the other stages with cervical node metastases between 10 and 20%. In the majority of the locally advanced cases, a combined therapy with irradiation and surgery should be performed. In cases with multiple or fixed neck nodes, a combined therapy should be performed. In cases with a single neck node metastasis which is not too large, radiotherapy mostly is sufficient. In most cases with TxN0, prophylactic neck irradiation is indicated.


Using empirical likelihood to combine data: application to food risk assessment.
This article introduces an original methodology based on empirical likelihood, which aims at combining different food contamination and consumption surveys to provide risk managers with a risk measure, taking into account all the available information. This risk index is defined as the probability that exposure to a contaminant exceeds a safe dose. It is naturally expressed as a nonlinear functional of the different consumption and contamination distributions, more precisely as a generalized U-statistic. This nonlinearity and the huge size of the data sets make direct computation of the problem unfeasible. Using linearization techniques and incomplete versions of the U-statistic, a tractable "approximated" empirical likelihood program is solved yielding asymptotic confidence intervals for the risk index. An alternative "Euclidean likelihood program" is also considered, replacing the Kullback-Leibler distance involved in the empirical likelihood by the Euclidean distance. Both methodologies are tested on simulated data and applied to assess the risk due to the presence of methyl mercury in fish and other seafood.


Cloning and characterization of the 5-aminolevulinate synthase gene(s) from Rhodobacter sphaeroides.
The 5-aminolevulinate synthase gene (hemA) from Rhizobium meliloti was used to probe a genomic lambda bank derived from Rhodobacter sphaeroides DNA. Two phage clones were found to bear homology to the Rhizobium probe. Southern hybridization analysis of the two lambda phage clones, which we designated lambda Hem 10 and lambda Hem 12, showed that the homology to the Rhizobium hemA gene was localized to a 3.1-kb SalI fragment derived from lambda Hem 10 and a 7.0-kb SalI fragment derived from lambda Hem 12. Each of the SalI fragments was subsequently cloned into the multiple cloning site of pUC19 in both orientations relative to the lac promoter. Restriction analysis confirmed that each SalI fragment was unique. It was also shown from Southern hybridization analysis that the regions of homology within each of the R. sphaeroides restriction fragments and the Rhizobium probe were different. Further, we have tentatively concluded that each R. sphaeroides hemA gene shows a relatively low degree of homology to the other. Data obtained from in vitro transcription-translation studies in a homologous R. sphaeroides cell-free system, and complementation of hemA mutations of both Escherichia coli and R. sphaeroides by either of the putative hemA clones suggested the presence of a gene encoding 5-aminolevulinate synthase on each DNA sequence. The fact that 5-aminolevulinate synthase activity could be demonstrated in mutant strains complemented in trans with either cloned DNA fragment further supported this conclusion.


Dynamics of the O(3P) + C2H4 reaction: identification of five primary product channels (vinoxy, acetyl, methyl, methylene, and ketene) and branching ratios by the crossed molecular beam technique with soft electron ionization.
The crossed molecular beam scattering technique with soft electron ionization (EI) is used to disentangle the complex dynamics of the polyatomic O(3P) + C2H4 reaction, which is of great relevance in combustion and atmospheric chemistry. Exploiting the newly developed capability of attaining universal product detection by using soft EI, at a collision energy of 54.0 kJ mol(-1), five different primary products have been identified, which correspond to the five exoergic competing channels leading to CH2CHO(vinoxy) + H, CH3CO(acetyl) + H, CH3(methyl) + HCO(formyl), CH2(methylene) + HCHO(formaldehyde), and CH2CO(ketene) + H2. From laboratory product angular and velocity distributions, center-of-mass product angular and translational energy distributions and the relative branching ratios for each channel have been obtained, affording an unprecedented characterization of this important reaction.


Unusual "cavity-in-cavity" appearance of pulmonary aspergilloma.
The classic radiographic appearance of pulmonary aspergilloma is a solid, round, often mobile intracavitary mass. We describe an unusual "cavity-in-cavity" appearance seen on conventional roentgenograms and computed tomography studies in two patients with confirmed pulmonary aspergilloma. We are not aware of any previous report describing such appearance based on literature search.


Colposcopic and histopathologic evaluation of women participating in population-based screening for human papillomavirus deoxyribonucleic acid persistence.
Evaluation of colposcopic and histopathological findings in women screened for cervical human papillomavirus deoxyribonucleic acid persistence. A total of 12 527 women, aged 32 to 38 years old, attending the population-based cervical cancer screening program in Sweden were randomized 1:1 to mock testing or human papillomavirus deoxyribonucleic acid testing by general primer 5+/6+ polymerase chain reaction and subsequent typing. Human papillomavirus deoxyribonucleic acid-positive women with a normal Papanicolaou smear (n=341) and an equal number from the control group were human papillomavirus tested on average 19 months later. One hundred nineteen women with type-specific human papillomavirus persistence and 111 controls were referred to colposcopy, and 84.8% attended. Histopathology from colposcopically directed biopsies confirmed cervical intraepithelial neoplasia grade 2 or 3 in 28 of 100 of the women with human papillomavirus deoxyribonucleic acid persistence and in 2 of 95 among controls. Among women with normal Papanicolaou smear attending population-based screening, the positive predictive value of human papillomavirus deoxyribonucleic acid persistence for detection of biopsy-confirmed cervical intraepithelial neoplasia 2 or 3 was 29%.


Serum and gene expression levels of leptin and adiponectin in rats susceptible or resistant to diet-induced obesity.
The aim of the present study was to identify the role of leptin and adiponectin in the development of resistance or susceptibility to diet-induced obesity in rats. For this purpose, male Wistar rats were fed with standard laboratory diet (control group) or cafeteria diet. After 15 days, two groups of rats with different response respect to the cafeteria diet were identified, and were assigned as diet-induced obesity (DIO) and diet resistant (DR) rats. The high-fat diet induced a very significant increase in both body and fat mass weight in DIO group. However, DR rats, gained even less weight than control-fed animals. Food intake was increased in cafeteria-fed rats (both DIO and DR) in comparison to control group; but hyperphagia was higher in DIO rats. In addition, feed efficiency (the ratio of weight gained to calories consumed) was significantly decreased in DR as compared to DIO rats. Regarding leptin, a significant increase in both adipose tissue gene expression and serum levels was observed in DIO rats in comparison with other groups (control and DR). A significant increase in both adiponectin circulating levels and adipose tissue mRNA expression was also observed in DIO animals as compared with the other groups. These data suggest that the susceptibility to obesity of DIO rats might be secondary, at least in part, to an earlier development of leptin resistance, which could lead to alterations in food intake (hyperphagia) and energetic metabolism. However, neither changes in leptin or adiponectin seem to be involved in the adaptive mechanisms that confer resistance to high fat intake.


Redefining periodic patterns on electroencephalograms of patients with sporadic Creutzfeldt-Jakob disease.
We aimed to redefine various periodic patterns (PPs) observed on electroencephalography (EEG) in patients with sporadic Creutzfeldt-Jakob disease (sCJD) using the American Clinical Neurophysiology Society's (ACNS) Criteria. We analyzed EEG data of 23 patients with sCJD were admitted to two university hospitals between August 2005 and September 2015. We classified PPs on EEG data into three types: irregular periodic discharges (PDs) with superimposed rhythmic activities, appearing at a median of 8weeks after onset (w.a.o.); rhythmic sharp-and-wave, at a median of 11w.a.o.; and PDs with biphasic or triphasic morphology, at a median of 17w.a.o. Of 16 patients presenting with PPs, 14 had widespread lesions in both cortical and subcortical areas with clinical stage III at admission, and shorter time intervals for admission to hospital from disease onset than patients without PPs (Patients with PP, 11.6±12.2weeks; without PP, 18.2±8.3weeks; p=0.033). PPs largely presented as three types at different stages of disease progression, and patients who had PPs had more wide spread lesions and rapid disease progression. Our redefinition of PPs demonstrated on EEG using the ACNS criteria may contribute to further understanding of the pathological mechanisms of sCJD, and PPs might be a predictive factor of a rapid sCJD progression.


Dose optimization of lithium to increase the uptake and retention of I-131 in rat thyroid.
The present study was undertaken to optimize the dose of lithium, with an aim to increase the retention of I-131 in the thyroid follicles while maintaining the euthyroid state. 24 female Wistar rats weighing 110 ± 20 g were segregated into four groups. Animals in group I were fed standard laboratory feed and water throughout the period of experimentation. Animals in group II, III and IV were additionally fed with lithium in the form of lithium carbonate orally, at a dose of 10 mg/kg body weight, 20 mg/kg body weight, 30 mg/kg body weight respectively. The dose of lithium was started 1 week prior to radioiodine administration and continued thereafter for another 8 days. After 7 days of lithium treatment, 0.48 MBq of carrier-free I-131 was injected intraperitoneally into each rat, of the four groups. I-131 thyroidal uptake and biokinetics, as well as serum TSH, T3, T4 levels were estimated in all the treatment groups. A significant increase in the thyroid and whole body counts was observed after 4 and 24 h of I-131 aministration in lithium treated rats, compared to control animals. An increase in thyroidal effective t1/2 and serum TSH levels, along with decrease in the levels of serum T3 and T4 was observed with a dose of 20 mg/kg or higher. In Conclusion, a Lithium dose of 10 mg/kg body weight in rats could increase the uptake of I-131 in the thyroid, without disturbing the control circulating levels of thyroid hormones.


Augmentation of cytotoxic responses by prostaglandin E2.
The cytotoxic-T-lymphocyte activity in the spleen cells after in vivo immunization of C3H mice with allogeneic spleen cells ip was consistently very weak. Substantial cytotoxic responses were obtained, however, when prostaglandins (PGE2, PGE1, or PGI2) were injected ip together with or prior to the immunization. An augmentation of cytotoxic responses against allogeneic stimulator cells was also observed in mixed lymphocyte cultures which were provided with an interleukin 2-containing helper factor. This augmentation was observed when PGE2 was added at the start of the culture but not if added 1 day later. Indomethacin was found to be suppressive in these cultures.


The A20 cDNA induced by tumor necrosis factor alpha encodes a novel type of zinc finger protein.
We have previously identified in endothelial cells a gene product designated A20, that is rapidly and profoundly induced by tumor necrosis factor alpha (Dixit, V. M., Green, S., Sarma, V., Holzman, L. B., Wolf, F. W., O'Rourke, K., Ward, P. A., Prochownik, E. V., and Marks, R. M. (1990) J. Biol. Chem. 265, 2973-2978). Isolation of the full-length cDNA followed by sequence analysis has revealed that A20 codes for a novel zinc finger protein. Structural features suggest that this putative protein, which contains multiple Cys2/Cys2 finger motifs, defines a novel class of zinc finger proteins. Southern analysis supports the existence of other members of the class.


Lysosomal deposition of Abeta in cultures of brain vascular smooth muscle cells is enhanced by iron.
Recently, we found that brain vascular smooth muscle cells from Tg2576 mice over-expressed the APP transgene in culture, secreted amyloid-beta peptide (Abeta) and accumulated Abeta intracellularly. Now we detected this intracellular Abeta inside lysosomes, which were also rich in C-terminal domain of APP, but not in endoplasmic reticulum, Golgi apparatus, or trans-Golgi network. Treatment of cultures with ferrous ions (50-150 microM) increased the proportion of muscle cells with Abeta immunoreactive granules and the amounts of intracellular Abeta1-40 and Abeta1-42 in a dose-dependent manner. This increase of intracellular Abeta1-40 by iron was inhibited by alpha-tocopherol, but not by a water-soluble antioxidant melatonin. The increase of intracellular Abeta1-42 by iron was not inhibited by alpha-tocopherol or melatonin. Cell treatment with iron did not alter the lysosomal localization of Abeta immunoreactivity. Cell treatment with iron (II and III), copper (II), zinc (II) and aluminum (III) increased cellular levels of carbonyls. However, the effect of zinc on Abeta accumulation in cultures was weak, and there were no effects of copper and aluminum. The data suggest that iron may be the factor that triggers vascular amyloidosis. Lysosomal accumulation of APP and Abeta initiates deposition of amyloid in blood vessels in Tg2576 mice.


Targeted deletion of ecto-5'-nucleotidase results in retention of inosine monophosphate content in postmortem muscle of medaka (Oryzias latipes).
Inosine monophosphate (IMP) is an important indicator of meat freshness and contributes to its umami taste. An attractive strategy for enhancing umami is to suppress the IMP-degrading activity and increase the IMP content in the skeletal muscle through genome editing technology using the CRISPR-Cas9 system. However, the molecular mechanisms underlying IMP degradation remain unclear. We cloned two ecto-5'-nucleotidase genes, designated as ecto-5'-nucleotidase-a (nt5ea) and ecto-5'-nucleotidase-b (nt5eb), from medaka (Oryzias latipes), a vertebrate model organism. Expression analysis using embryos showed that nt5ea or nt5eb overexpression remarkably upregulated IMP degradation, and that the IMP-degrading activity was higher in Nt5ea than in Nt5eb. Furthermore, we established frame-shifted or large deletion (lacking nt5ea or nt5eb locus) mutant strains and assayed the effects of gene disruptions on the amount of IMP in skeletal muscle. The nt5ea-deficient medaka showed considerable higher levels of IMP at 48 h postmortem than did the wild-type fish. The nt5eb mutants also exhibited higher IMP contents than that in the wild types, but the increase was less than that in the nt5ea mutants. Our results demonstrated that nt5e is an important regulator of IMP levels in skeletal muscle and that its loss of function was effective in maintaining IMP content.


Diagnosis of neurinoma in the region of the jugular foramen--a case report.
Neurinomas in the region of the jugular foramen are extremely rare, are difficult to diagnose before surgery, and have seldom been described in the literature. The typical jugular foramen syndromes (Collet-Sicard syndrome, Vernet's paralysis) may be caused by many various disorders. The diagnostic problems are illustrated using one of our cases as an example. The differential diagnosis was made before surgery using currently available radiologic techniques. The tentative diagnosis "neurinoma of the hypoglossal nerve" was confirmed intraoperatively and histologically.


A comparative study of the characteristics of eIF-2 and eIF-2-ancillary factor activities from yeast Saccharomyces cerevisiae and rabbit reticulocytes.
The characteristics of yeast eukaryotic initiation factor 2 (eIF-2) and Co-eIF-2A have been studied and compared with those of the corresponding factors from rabbit reticulocytes. 1) Unlike eIF-2r, purified eIF-2y did not contain bound GDP. 2) Purified eIF-2y preparation contained GTPase activity and dephosphorylated GTP to GDP. 3) An anti-eIF-2r preparation which predominantly precipitated the gamma-subunit (Mr 54,000) of eIF-2r also precipitated the larger subunit (Mr 54,000) of eIF-2y. 4) Unlike eIF-2r, ternary complex formation by eIF-2y was not inhibited by Mg2+. 5) Both Co-eIF-2A20y and Co-eIF-2r significantly enhanced Met-tRNAf binding to eIF-2y and, again, Mg2+ did not have any effect on this stimulated Met-tRNAf binding to eIF-2y. 6) Both Co-eIF-2A20y and Co-eIF-2r were similarly effective in stimulating Met-tRNAf binding to eIF-2r in the absence of Mg2+. However, in the presence of Mg2+, Co-eIF-2A20y was significantly less effective than Co-eIF-2r as Co-eIF-2A20y did not promote displacement of GDP from eIF-2r X GDP. 7) eIF-2y bound [3H]GDP and this binding was significantly enhanced in the presence of Mg2+. Also, [3H]GDP in the preformed eIF-2y X [3H]GDP complex was rapidly exchanged with exogenously added unlabeled GDP in the presence of Mg2+. Co-eIF-2A20y had no effect on GDP binding to eIF-2y nor on GDP exchange reactions. 8) Reticulocyte heme-regulated protein synthesis inhibitor, which phosphorylated almost completely (in excess of 80%) the alpha-subunit (Mr 38,000) of eIF-2r, also phosphorylated similarly the smaller subunit (Mr 36,000) of eIF-2y. However, such phosphorylation had no significant effect on ternary complex formation, GDP binding, and GDP exchange reactions.


Partial loss of arcuate kisspeptin neurons in female rats stimulates luteinizing hormone and decreases prolactin secretion induced by estradiol.
Kisspeptin, neurokinin, and dynorphin (KNDy) neurons in the arcuate nucleus (ARC) control luteinizing hormone (LH) and prolactin (PRL) release, although their role in conveying the effects of estradiol (E2 ) to these hormones is not well understood. We performed a longitudinal evaluation of female rats in which KNDy neurons were ablated using a neurokinin-3 receptor agonist conjugated with saporin (NK3-SAP) to investigate the impact of the reduction of KNDy neurons on the E2 regulation of gonadal and PRL axes. NK3-SAP rats, bearing a moderate loss of ARC kisspeptin-immunoreactive (-IR) neurons (50%-90%), displayed irregular estrous cycles but essentially unaltered follicular development and a normal number of corpora lutea. Rats were then ovariectomized (OVX) and treated with a positive-feedback dose of E2 (OVX + E2 ). LH and PRL were measured in the tail blood by an enzyme-linked immunosorbent assay. The E2 -induced LH surge was amplified, whereas the PRL rise was decreased in NK3-SAP rats compared to Blank-SAP control. After 10 days of no hormonal treatment, basal LH levels were equally elevated in NK3-SAP and controls. Tyrosine hydroxylase (TH) phosphorylation in the median eminence, in turn, was increased in NK3-SAP rats, with no change in the number of ARC TH-IR neurons. Thus, KNDy neurons exert concurrent and opposite roles in the E2 -induced surges of LH and PRL. The partial loss of KNDy neurons disrupts ovarian cyclicity but does not preclude ovulation, consistent with the disinhibition of the LH preovulatory surge. Conversely, KNDy neurons tonically inhibit the enzymatic activity of tuberoinfundibular dopaminergic neurons, which appears to facilitate PRL release in response to E2 .


Prediction of Curve Progression in Idiopathic Scoliosis: Validation of the Sanders Skeletal Maturity Staging System.
Retrospective case series. This study aimed to validate the Sanders Skeletal Maturity Staging System and to assess its correlation to curve progression in idiopathic scoliosis. The Sanders Skeletal Maturity Staging System has been used to predict curve progression in idiopathic scoliosis. This study intended to validate that initial study with a larger sample size. We retrospectively reviewed 1100 consecutive patients with idiopathic scoliosis between 2005 and 2011. Girls aged 8 to 14 years (<2 yr postmenarche) and boys aged 10 to 16 years who had obtained at least 1 hand and spine radiograph on the same day for evaluation of skeletal age and scoliosis curve magnitude were followed to skeletal maturity (Risser stage 5 or fully capped Risser stage 4), curve progression to 50° or greater, or spinal fusion. Patients with nonidiopathic curves were excluded. There were 161 patients: 131 girls (12.3 ± 1.2 yr) and 30 boys (13.9 ± 1.1 yr). The distribution of patients within Sanders stage (SS) 1 through 7 was 7, 28, 41, 45, 7, 31, and 2 patients, respectively; modified Lenke curve types 1 to 6 were 26, 12, 63, 5, 38, and 17 patients, respectively. All patients in SS2 with initial Cobb angles of 25° or greater progressed, and patients in SS1 and SS3 with initial Cobb angles of 35° or greater progressed. Similarly, all patients with initial Cobb angles of 40° or greater progressed except those in SS7. Conversely, none of the patients with initial Cobb angles of 15° or less or those in SS5, SS6, and SS7 with initial Cobb angles of 30° or less progressed. Predictive progression of 67%, 50%, 43%, 27%, and 60% was observed for subgroups SS1/30°, SS2/20°, SS3/30°, SS4/30°, and SS6/35° respectively. This larger cohort shows a strong predictive correlation between SS and initial Cobb angle for probability of curve progression in idiopathic scoliosis. 3.


A simpler analysis for the measurement of second virial coefficients by self-interaction chromatography.
We describe a thermodynamic approach that supports the adoption of a simplified procedure for the determination of protein second virial coefficients (B(2)) by self-interaction chromatography. Its major advantage over the original method is a decrease in the number of parameters to which magnitudes must be assigned for the determination of B(2). Improved correlation of virial coefficients obtained by the chromatographic procedure with those obtained by light scattering is achieved by taking into account the twofold larger magnitudes of the former because of the experimental distinction between free and immobilized protein molecules in self-interaction chromatography.


Controllable synthesis of MnO2/polyaniline nanocomposite and its electrochemical capacitive property.
Polyaniline (PANI) and MnO2/PANI composites are simply fabricated by one-step interfacial polymerization. The morphologies and components of MnO2/PANI composites are modulated by changing the pH of the solution. Formation procedure and capacitive property of the products are investigated by XRD, FTIR, TEM, and electrochemical techniques. We demonstrate that MnO2 as an intermedia material plays a key role in the formation of sample structures. The MnO2/PANI composites exhibit good cycling stability as well as a high capacitance close to 207 F g-1. Samples fabricated with the facile one-step method are also expected to be adopted in other field such as catalysis, lithium ion battery, and biosensor.


Detection of single mammalian cells by high-resolution magnetic resonance imaging.
This study reports the detection of single mammalian cells, specifically T cells (T lymphocytes) labeled with dextran-coated superparamagnetic iron oxide particles, using magnetic resonance microscopy. Size amplification due to sequestration of the superparamagnetic particles in vacuoles enhances contrast in localized areas in high-resolution magnetic resonance imaging. Magnetic resonance images of samples containing differing concentrations of T cells embedded in 3% gelatin show a number of dark regions due to the superparamagnetic iron oxide particles, consistent with the number predicted by transmission electron microscopy. Colabeling of T cell samples with a fluorescent dye leads to strong correlations between magnetic resonance and fluorescence microscopic images, showing the presence of the superparamagnetic iron oxide particles at the cell site. This result lays the foundation for our approach to tracking the movement of a specific cell type in live animals and humans.


Pituitary alcohol injection for cancer pain. Use in a district general hospital.
The use of trans-sphenoidal pituitary alcohol injection was assessed in a district general hospital for the treatment of diffuse bilateral cancer pain in 26 patients. Fourteen patients received a total of 43 injections. Hormonal and non-hormonal dependent cancers were treated, with complete pain relief in ten patients and partial relief in a further seven patients. The duration of relief was variable, and usually less than 6 weeks, although two patients had complete pain relief for 5 and 7 months respectively. No cases of meningitis or cerebrospinal fluid rhinorrhoea occurred. Nasal antiseptics and vasoconstrictors were not used. Temporary pupil dilatation occurred in eight patients, of whom one had prolonged blurred vision for small print. Transient diabetes insipidus was common (53%), although only five patients needed permanent treatment. Death from their disease occurred in 65% of patients within 3 months. Survival was shorter than in series from specialist centres, suggesting later referral for pain relief. The technique is an inexpensive and valid form of treatment for cancer pain in a district hospital, but should not be undertaken lightly in view of the possible complications.


The web factor in cholangitis.
Choledochoscopy demonstrates that stone-related suppurative cholangitis has the pathologic characteristics of a multiloculated intraductal abscess. The fibrin web compartmentalizes the duct, traps calculi, and limits spontaneous extrusion. The stones act as foreign bodies and stasis maintains the bacterobilia. This is the anatomic basis and rationale for early direct surgical drainage as the definite effective treatment.


Separation of 1-phenyl-3-methyl-5-pyrazolone derivatives of monosaccharides by capillary electrochromatography.
1-Phenyl-3-methyl-5-pyrazolone (PMP) derivatives of component monosaccharides in glycoproteins (fucose, galactose, mannose, N-acetylgalactosamine and N-acetylglucosamine) and epimeric aldopentoses (arabinose, lyxose, ribose and xylose) were well separated from each other by capillary electrochromatography on a Hypersil ODS column with a mixture of 50 mM N-(2-hydroxyethyl)piperazine-2'-(2-ethanesulfonic acid) buffer, pH 6.0 to approximately 6.3, and acetonitrile (2.2:1 v/v) as eluent. The elution of these compounds showed relatively strong dependence on the pH and concentration of the buffer salts contained in the eluent, as compared to the elution by pressure-driven high-performance liquid chromatography (HPLC) on the same stationary phase, but separation of PMP-monosaccharides was better than that by HPLC. Retention times of PMP-monosaccharides were highly reproducible with a relative standard deviation (RSD) of approximately 0.6%, and quantification with an RSD less than 5% could be achieved using 3-O-methylglucose as an internal standard.


Detection of metastatic thyroid carcinoma after the administration of a therapeutic dose of 131-iodine.
A patient with thyroid cancer is described whose metastases were detected only after the administration of a large therapeutic dose of 131 Iodine. This suggests that very high doses of 131 Iodine might permit diagnoses of metastatic lesions not otherwise detected. This phenomenon may be explained by the improved counting statistics which occur with the use of a higher dose of the isotope. In such cases, exceedingly high doses of 131 Iodine might be necessary for successful tumor ablation.


Characterization of four urotensin II receptors (UTS2Rs) in chickens.
Urotensin II receptor (UTS2R) is suggested to mediate the actions of urotensin II (UTS2) and UTS2-related peptide (URP, also called UTS2B) in mammals. However, the information regarding the gene structure, functionality and tissue expression of UTS2/URP receptor remains largely unknown in non-mammalian vertebrates including birds. In this study, using RACE-PCR, we cloned the full-length cDNAs of four chicken UTS2/URP receptors and designated them as cUTS2R1, cUTS2R2, cUTS2R3 and cUTS2R5 respectively, according to their evolutionary origin. The cloned cUTS2R1, cUTS2R2, cUTS2R3 and cUTS2R5 are predicted to encode 7-transmembrane receptors of 382, 343, 331 and 363 amino acids respectively, which show 50-66 % amino acid sequence identity with human UTS2R. Using cell-based luciferase reporter assays and Western blot, we demonstrated that chicken UTS2Rs expressed in HEK293 cells could be effectively activated by synthetic chicken UTS2-12, UTS2-17 and URP peptides, and their activation can elevate intracellular calcium concentration and activate MAPK/ERK signaling cascade, indicating that the four UTS2Rs are functional and capable of mediating UTS2/URP actions in chickens. Quantitative real-time PCR revealed that the four receptors are widely, but differentially, expressed in adult chicken tissues, while cUTS2 and cURP are highly expressed in the hindbrain and spinal cord, and moderately/weakly expressed in other tissues examined including the spleen and gonads. Taken together, our data provide first piece of evidence that all four UTS2Rs are functional in an avian species and help to reveal the conserved roles of UTS2R signaling across vertebrates.


The effects of LXR agonist T0901317 and LXR antagonist GSK2033 on morphogenesis and lipid properties in full thickness skin models.
Full thickness models (FTMs) are 3D-cultured human skin models that mimic many aspects of native human skin (NHS). However, their stratum corneum (SC) lipid composition differs from NHS causing a reduced skin barrier. The most pronounced differences in lipid composition are a reduction in lipid chain length and increased monounsaturated lipids. The liver-X-receptor (LXR) activates the monounsaturated lipid synthesis via stearoyl-CoA desaturase-1 (SCD-1). Therefore, the aim was to improve the SC lipid synthesis of FTMs by LXR deactivation. This was achieved by supplementing culture medium with LXR antagonist GSK2033. LXR agonist T0901317 was added for comparison. Subsequently, epidermal morphogenesis, lipid composition, lipid organization and the barrier functionality of these FTMs were assessed. We demonstrate that LXR deactivation resulted in a lipid composition with increased overall chain lengths and reduced levels of monounsaturation, whereas LXR activation increased the amount of monounsaturated lipids and led to a reduction in the overall chain length. However, these changes did not affect the barrier functionality. In conclusion, LXR deactivation led to the development of FTMs with improved lipid properties, which mimic the lipid composition of NHS more closely. These novel findings may contribute to design interventions to normalize SC lipid composition of atopic dermatitis patients.


[Scleritis and episcleritis].
Scleritis and episcleritis are rare ocular inflammatory diseases but deserve to be known by internists because of their frequent association with systemic autoimmune diseases. It is important to distinguish them between because their prognosis, therapeutic management and potential complications are very different. Episcleritis represents a superficial ocular inflammation with usually benign visual prognosis, no complication with local treatment, and is associated with a systemic autoimmune disease in rare cases. In contrast, scleritis is a potentially serious ophthalmological condition that can threaten the visual prognosis in the absence of appropriate systemic treatment. It is associated with an underlying disease in 40-50% of cases, in particular a systemic autoimmune disease (25-35% of cases) or an infectious cause (5-10% of cases). Rheumatoid arthritis and systemic vasculitides, particularly antineutrophil cytoplasmic antibody (ANCA)-associated vasculitides, are the main autoimmune causes of scleritis and episcleritis. Scleritis can reveal the underlying autoimmune disease and requires systematic etiological investigations. Aggressive, complicated, refractory forms or those associated with a systemic autoimmune disease require glucocorticoids or even immunosuppressants, and close collaboration between ophthalmologists and internists is required. The development of biologic agents offers new effective therapeutic tools in the management of these difficult cases.


Angiotensin II receptor-independent antiinflammatory and antiaggregatory properties of losartan: role of the active metabolite EXP3179.
Angiotensin II (Ang II) type 1 receptor (AT(1)) antagonists such as losartan (LOS) are widely used for the treatment of hypertension and elicit antiinflammatory and antiaggregatory in vitro and in patients, although the underlying mechanism are unclear. Following computer-based molecule similarity, we proposed that on cytochrome-P450 degradation, the LOS metabolite EXP3179 is generated, which shows molecule homology to indomethacin, a cyclooxygenase inhibitor with antiinflammatory and antiaggregatory properties. Subsequently, serum-levels of EXP3179 were determined for 8 hours in patients receiving a single oral dose of 100 mg LOS. High-performance liquid chromatography followed by liquid chromatography-mass spectrometry (GC-MS) [corrected] from serum samples revealed a maximum of 10(-7) mol/L for EXP3179 peaking between 3 to 4 hours. The increase in serum-EXP3179 levels was associated with a significant reduction in platelet aggregation in vivo (-35+/-4%, P<0.001 versus control). EXP3179 generation was investigated in a chemical reaction mimicking the liver cytochrome-P450-dependent LOS-degradation and human endothelial cells were exposed to Ang II or lipopolysaccharides (LPS) in the presence of EXP3179 (10(-7) mol/L). LPS- and Ang II-induced COX-2 transcription was abolished by EXP3179. Moreover, EXP3179 significantly reduced Ang II- and LPS-induced formation of prostaglandin F2alpha as determined by GC-MS [corrected]. Thus, antiinflammatory properties of LOS are mediated via its EXP3179 metabolite by abolishing COX-2 mRNA upregulation and COX-dependent TXA2 and PGF2alpha generation. Serum levels of EXP3179 are detectable in patients in concentrations that exhibit antiinflammatory and antiaggregatory properties in vitro.


Reliability of intraocular pressure measurements after myopic excimer photorefractive keratectomy.
To determine the reliability of intraocular pressure (IOP) measurements by Goldmann applanation tonometry and pneumotonometry in eyes treated with excimer myopic photorefractive keratectomy (PRK). A prospective case series. Forty consecutive eyes treated with PRK were evaluated. Central and peripheral corneal Goldmann tonometry and pneumotonometry measurements were done before surgery, at 1 week, and at 1 and 3 months after surgery. The IOP by Goldmann tonometry from the central cornea was significantly lower than the peripheral IOP; however, there was no difference between IOP measured from central and peripheral corneas by pneumotonometry, which, in turn, correlated with peripheral Goldmann measurements. There was a trend, but not a statistically significant correlation, between the spherical equivalent of the treatment and the amount of decrease in central Goldmann IOP. Pneumotonometry measures the IOP reliably after PRK from all parts of the cornea, whereas central Goldmann tonometry underestimates the IOP by 2.40+/-1.23 mmHg.


[Not Available].
Two toxicity tests (pollen tube growth test and methylen blue-assay) were examined regarding their sensitivity towards certain chosen mycotoxins. The IC50 values of the PTGT were in a concentration range between 0,01 μg/ml and 11,18 μg/ml, of the MB-assay in a concentration range between 0,003 μg/ml and 17,91 μg/ml. Both tests can be used to record toxigenic fungal wild strains in mould-infested apartments. Independently from the well-known immunochemical methods or HPLC/GC-MS analysis must be used for an exact determination of the kind and the content of the mycotoxins.


Nutritional effects on NF-kappaB/IkappaB activity in weight-gaining and -losing rats.
We had previously reported metabolic abnormalities in TNF-treated weight-gaining (WG) rats, associated with increased soluble and membrane TNF receptors. On the basis of these findings, we hypothesized that stabilization of TNF in circulation in WG animals by these receptors augmented the action of TNF. TNF influences cellular function through the activation of NF-kappaB, by its release from IkappaBalpha and translocation to the nucleus where it regulates gene transcription. This study was designed to determine whether NF-kappaB binding was increased in WG rats in comparison with weight-losing (WL) rats. Twenty two male Wistar rats weighing 200-220 g were fed a liquid-defined formula diet for 10 days and divided as follows: (i) WG rats (n = 11) receiving an energy density of 0.35 MJ per 60 ml and (ii) WL rats (n = 11) receiving an energy density of 0.09 MJ per 60 ml. Nuclear extracts from heart, liver, lung, spleen and gastrocnemius were used for NF-kappaB measurements by electrophoretic mobility shift assay. Tissue lysates from heart and lung were used for IkappaBalpha measurements by Western blot analysis. There was increased NF-kappaB binding in the lung and heart extracts of the WG rats as compared with WL rats. There was no difference in the binding activity between WG and WL rats in the spleen, liver and gastrocnemius extracts. In lung and heart lysates, IkappaBalpha protein expression was decreased in WG rats as compared with the WL rats. We conclude that there is increased organ-specific TNF activity in WG rats because of increased binding of NF-kappaB to the nucleus and reduced IkappaBalpha levels in the lung and heart. The findings are consistent with increased TNF activity resulting in the release of NF-kappaB from the NF-kappaB/IkappaBalpha complex and translocation to the nucleus. The nuclear translocation of NF-kappaB observed in the heart and lung suggests that nutrition may influence the action of TNF in these vital organs in sepsis. This finding is potentially of great significance, as pulmonary and cardiac failure are the main causes of death in sepsis.


Rapid quantitative detection of the discrepant compounds in differently processed Curcumae Rhizoma products by FT-NIR combined with VCPA-GA technology.
Curcumae Rhizoma (CR) and vinegar processed Curcumae Rhizoma (PCR) are common medicinal materials widely used in clinical practice in China. There are sliced CR (SCR) and two kinds of PCR products which are processed by different methods: WPCR-prepared with the whole CR root boiled in vinegar and then sliced, and SPCR-prepared with the whole CR root steamed and sliced before boiled with vinegar. In this study, the feasibility of Fourier transform near infrared spectrum (FT-NIR) used to determine the main discrepant components of SCR, WPCR and SPCR were investigated. High performance liquid chromatography (HPLC) was used to identified five discrepant compounds in the three kinds of CR products-curzerene, curcumenol, curdione, furanodienone and demethoxycurin. Pretreatment of NIR qualitative data by different methods revealed that the second derivative in combination with 9 points of Savitzky-Golay smooth (2D9S) could accurately distinguish SCR, SPCR and WPCR from each other, and the discrimination ability was improved significantly by wavebands selection. Then a model with great accuracy was established by combining with wavebands selection and partial least squares regression (PLSR). Compared with the competitive adaptive reweighted sampling (CARS) selection method, 2D9S- variable combination population analysis (VCPA)-Genetic algorithm (GA)-PLSR model was evidently more accurate in prediction of the content of curzerene, curcumenol, curdione and furanodienone, with an R2p of 0.9558, 0.9129, 0.9098 and 0.9350, as well as a ratio of performance to deviation (RPD) of 4.8454, 3.4640, 3.3020 and 4.0082, respectively. Whereas, the content of demethoxycurin failed to be well predicted. The correlation analysis revealed that the results of wavebands selection were consistent with the trend of changes in the content of these target compounds and the findings of NIR absorption analysis, and the characteristic chemical bonds of these compounds corresponded to the areas with significant correlation in the heat map. It can be concluded that the NIR system, combined with appropriate variable selection and linear regression method, can precisely distinguish SCR, SPCR and WPCR from each other, and can accurately and rapidly determine the four discrepant compounds in the three CR products, suggesting a potential of being routinely used for a more diversified analysis in medicinal herbs study.


Intracranial trigeminal nerve rhabdomyoma/choristoma in a child: a case report and discussion of possible histogenesis.
Rhabdomyomas are rare tumors that usually arise within the heart, orocervical, or vulvovaginal regions. The cardiac tumors have a characteristic immature morphology, occur often in association with tuberous sclerosis, and are regarded as hamartomas rather than true neoplasms. The histogenesis of the extracardiac tumors and their true neoplastic nature are matters of controversy. We report the first case of a rhabdomyoma located inside the cranium. The intimate association with the mandibular division of the trigeminal nerve, the normal embryogenesis of the craniofacial muscles, and animal homograft and xenograft experiments provide a framework for considering this tumor, and possibly other rhabdomyomas, as a choristoma/hamartoma rather than a true neoplasm.


A seasonal difference of daily energy expenditure in a free-living subterranean rodent, the silvery mole-rat (Heliophobius argenteocinereus; Bathyergidae).
In seasonal climatic regimes, animals have to deal with changing environmental conditions. It is reasonable to expect that seasonal changes are reflected in animal overall energetics. The relation between daily energy expenditure (DEE) and seasonally variable ecological determinants has been studied in many free-living small mammals; however with inconsistent results. Subterranean mammals, i.e. fossorial (burrowing) mammals which live and forage underground, live in a seasonally and diurnally thermally stable environment and represent a suitable model to test seasonality in DEE in respect to seasonal changes, particularly those in soil characteristics and access to food supply. Both factors are affected by seasonal rainfall and are supposed to fundamentally determine activity of belowground dwellers. These ecological constraints are pronounced in some tropical regions, where two distinct periods, dry and rainy seasons, regularly alternate. To explore how a tropical mammal responds to an abrupt environmental change, we determined DEE, resting metabolic rate (RMR) and sustained metabolic scope (SusMS) in a solitary subterranean rodent, the silvery mole-rat, at the end of dry season and the onset of rainy season. Whereas RMR did not differ between both periods, mole-rats had 1.4 times higher DEE and SusMS after the first heavy rains. These findings suggest that rainfall is an important environmental factor responsible for higher energy expenditure in mole-rats, probably due to increased burrowing activity. SusMS in the silvery mole-rat is comparable to values in other bathyergids and all bathyergid values rank among the lowest SusMS found in endothermic vertebrates.