Zethrenes, extended p-quinodimethanes, and periacenes with a singlet biradical ground state.
Researchers have studied polycyclic aromatic hydrocarbons (PAHs) for more than 100 years, and most PAHs in the neutral state reported so far have a closed-shell electronic configuration in the ground state. However, recent studies have revealed that specific types of polycyclic hydrocarbons (PHs) could have a singlet biradical ground state and exhibit unique electronic, optical, and magnetic activities. With the appropriate stabilization, these new compounds could prove useful as molecular materials for organic electronics, nonlinear optics, organic spintronics, organic photovoltaics, and energy storage devices. However, before researchers can use these materials to design new devices, they need better methods to synthesize these molecules and a better understanding of the fundamental relationship between the structure and biradical character of these compounds and their physical properties. Their biradical character makes these compounds difficult to synthesize. These compounds are also challenging to physically characterize and require the use of various experimental techniques and theoretic methods to comprehensively describe their unique properties. In this Account, we will discuss the chemistry and physics of three types of PHs with a significant singlet biradical character, primarily developed in our group. These structures are zethrenes, Z-shaped quinoidal hydrocarbons; hydrocarbons that include a proaromatic extended p-quinodimethane unit; and periacenes, acenes fused in a peri-arrangement. We used a variety of synthetic methods to prepare these compounds and stabilized them using both thermodynamic and kinetic approaches. We probed their ground-state structures by electronic absorption, NMR, ESR, SQUID, Raman spectroscopy, and X-ray crystallography and also performed density functional theory calculations. We investigated the physical properties of these PHs using various experimental methods such as one-photon absorption, two-photon absorption, transient absorption spectroscopy, electrochemistry, and spectroelectrochemistry. These systematic studies revealed that aromaticity played a very important role in determining their singlet biradical character, which is critically related to both their physical properties and their chemical reactivity. In particular, we found that Clar's aromatic sextet rule, which is useful for the closed-shell PAHs, can also predict the relative biradical character of benzenoid PH-based singlet biradicaloids. Other factors, such as structural flexibility of the biradical and quinoid resonance forms and the participation of the substitution in the π-conjugation, also influence the biradical character. These molecular materials demonstrate a number of unique properties such as near-infrared absorption, redox amphotericity, large two-photon absorption cross section, short excited state lifetime, stimuli-responsive magnetic activity, and singlet fission, which suggests promise for future applications.


Experimental induction of bacillary hemoglobinuria in cattle.
An open-surgery technique for intrahepatic inoculation of Clostridium haemolyticum spores, suspended in calcium chloride as the hepatic debilitant, was used to produce bacillary hemoglobinuria in cattle. All calves (n=3) died of the disease, and the controls (n=2) given calcium chloride without spores survived. Clinical signs and gross pathologic changes produced by this method resembled those described for the disease in its natural form.


Enhancing Pore-Environment Complexity Using a Trapezoidal Linker: Toward Stepwise Assembly of Multivariate Quinary Metal-Organic Frameworks.
Multicomponent metal-organic frameworks (MOFs) promise the precise placement of synergistic functional groups with atomic-level precision, capable of promoting fascinating developments in basic sciences and applications. However, the complexity of multicomponent systems poses a challenge to their structural design and synthesis. Herein, we show that linkers of low symmetry can bring new opportunities to the construction of multicomponent MOFs. A carbazole-tetracarboxylate linker of C s point group symmetry was designed and combined with an 8-connected Zr6 cluster to generate a low-symmetry MOF, PCN-609. PCN-609 contains coordinatively unsaturated Zr sites arranged within a lattice with three crystallographically distinct pockets, which can accommodate linear linkers of different lengths. Sequential linker installation was carried out to postsynthetically insert three linear linkers into PCN-609, giving rise to a quinary MOF. Functionalization of each linker from the quinary MOF system creates multivariate pore environments with unprecedented complexity.


Predictors of Long-Term Outcome of Isolated Surgical Aortic Valve Replacement in Aortic Regurgitation With Reduced Left Ventricular Ejection Fraction and Extreme Left Ventricular Dilatation.
The management of severe aortic regurgitation (AR) in patients with reduced left ventricular function and extreme left ventricular dilatation presents a therapeutic dilemma. This study aims to assess risk factors of aortic valve replacement (AVR) for these particular population based on its performances. Two hundred twelve severe AR patients accompanied by left ventricular ejection fraction (LVEF) <50% and left ventricular end-diastolic dimension (LVEDD) ≥70 mm who underwent isolated AVR between January 2007 and December 2016 were identified retrospectively. Logistic regression and receiver operating characteristic were used to analyze prognostic indicators for in-hospital mortality while Kaplan-Meier analysis for long-term survival. Mean age was 56 ± 13 years with mean LVEF 40 ± 7% and LVEDD 78 ± 6 mm. In-hospital mortality rate was 7%, and survival rates at 5 and 10 years were 88 ± 4% and 73 ± 10%, respectively. Logistic regression analysis indicated in-hospital mortality was associated with preoperative age and LVEF. Receiver operating characteristic analysis showed LVEF = 35% was the best cut-off value at which to predict in-hospital death. Kaplan-Meier analysis revealed patients with markedly reduced LV function (LVEF <35%) had lower survival rates compared with other patients with moderate LV dysfunction (LVEF 36% to 50%) (1-, 5-, and 10-year: 90 ± 4%, 64 ± 7%, and 55 ± 14%, vs 97 ± 1%, 94 ± 3%, and 76 ± 7%, p <0.001). An age-matched analysis showed similar trend (p = 0.020). In Conclusion, AVR may be unsafe for severe AR patients with markedly reduced LV function (LVEF <35%) and extreme left ventricular dilatation (LVEDD >70 mm) due to poor postoperative early- and long-term outcomes.


Multiple signal input and output domains of the 160-kilodalton nuclear receptor coactivator proteins.
Members of the 160-kDa nuclear receptor coactivator family (p160 coactivators) bind to the conserved AF-2 activation function found in the hormone binding domains of nuclear receptors (NR) and are potent transcriptional coactivators for NRs. Here we report that the C-terminal region of p160 coactivators glucocorticoid receptor interacting protein 1 (GRIP1), steroid receptor coactivator 1 (SRC-1a), and SRC-1e binds the N-terminal AF-1 activation function of the androgen receptor (AR), and p160 coactivators can thereby enhance transcriptional activation by AR. While they all interact efficiently with AR AF-1, these same coactivators have vastly different binding strengths with and coactivator effects on AR AF-2. p160 activation domain AD1, which binds secondary coactivators CREB binding protein (CBP) and p300, was previously implicated as the principal domain for transmitting the activating signal to the transcription machinery. We identified a new highly conserved motif in the AD1 region which is important for CBP/p300 binding. Deletion of AD1 only partially reduced p160 coactivator function, due to signaling through AD2, another activation domain located at the C-terminal end of p160 coactivators. C-terminal coactivator fragments lacking AD1 but containing AD2 and the AR AF-1 binding site served as efficient coactivators for full-length AR and AR AF-1. The two signal input domains (one that binds NR AF-2 domains and one that binds AF-1 domains of some but not all NRs) and the two signal output domains (AD1 and AD2) of p160 coactivators played different relative roles for two different NRs: AR and thyroid hormone receptor.


CpG island methylator phenotype is a strong determinant of poor prognosis in neuroblastomas.
Neuroblastoma, one of the most common pediatric solid tumors, is characterized by two extreme disease courses, spontaneous regression and life-threatening progression. Here, we conducted a genome-wide search for differences in DNA methylation that distinguish between neuroblastomas of the two types. Three CpG islands (CGI) and two groups of CGIs were found to be methylated specifically in neuroblastomas with a poor prognosis. By quantitative analysis of 140 independent cases, methylation of all the five CGI (groups) was shown to be closely associated with each other, conforming to the CpG island methylator phenotype (CIMP) concept. The presence of CIMP was sensitively detected by methylation of the PCDHB CGIs and associated with significantly poor survival (hazard ratio, 22.1; 95% confidence interval, 5.3-93.4; P < 0.0001). Almost all cases with N-myc amplification (37 of 38 cases) exhibited CIMP. Even in 102 cases without N-myc amplification, the presence of CIMP (30 cases) strongly predicted poor survival (hazard ratio, 12.4; 95% confidence interval, 2.6-58.9; P = 0.002). Methylation of PCDHB CGIs, located in their gene bodies, did not suppress gene expression or induce histone modifications. However, CIMP was significantly associated with methylation of promoter CGIs of the RASSF1A and BLU tumor suppressor genes. The results showed that neuroblastomas with CIMP have a poor prognosis and suggested induction of silencing of important genes as an underlying mechanism.


Mesoporous magnetic silica particles modified with stimuli-responsive P(NIPAM-DMA) valve for controlled loading and release of biologically active molecules.
Mesoporous magnetic silica particles bearing a stimuli-responsive polymer valve were prepared and their performance as a microcapsule was evaluated. In this study, first, mesoporous magnetic iron oxide (Fe3O4) particles were prepared by a solvothermal method. Then, the magnetic particles were coated with silica and functionalized with vinyl groups using 3-(trimethoxysilyl)-propyl methacrylate (MPS). Subsequently, the Fe3O4/SiO2 composite particles grafted with MPS were used to carry out the seeded precipitation copolymerization of N-isopropylacrylamide (NIPAM) and 2,2-dimethylaminoethyl methacrylate (DMA). Here N,N'-methylenebisacrylamide (MBA) was used as a cross-linker. Brunauer-Emmett-Teller (BET) surface analysis suggested that the mesoporous structure was retained in the final Fe3O4/SiO2/P(NIPAM-DMA-MBA) composite hydrogel particles. The prepared Fe3O4/SiO2/P(NIPAM-DMA-MBA) composite hydrogel microspheres exhibited a pH-dependent volume phase transition. At lower pH values (<7), the inclusion of DMA shifted the volume phase transition to higher temperature because of the protonation of the tertiary amine groups. The composite hydrogel particles possessed a high saturation magnetization (51 emu g-1) and moved under the influence of an external magnetic field. The loading-release behaviour of these biologically active molecules suggested that a portion of the encapsulated guest molecules was released at a temperature below the lower critical solution temperature, LCST (<35 °C).


Living nano-micro fibrous woven fabric/hydrogel composite scaffolds for heart valve engineering.
Regeneration and repair of injured or diseased heart valves remains a clinical challenge. Tissue engineering provides a promising treatment approach to facilitate living heart valve repair and regeneration. Three-dimensional (3D) biomimetic scaffolds that possess heterogeneous and anisotropic features that approximate those of native heart valve tissue are beneficial to the successful in vitro development of tissue engineered heart valves (TEHV). Here we report the development and characterization of a novel composite scaffold consisting of nano- and micro-scale fibrous woven fabrics and 3D hydrogels by using textile techniques combined with bioactive hydrogel formation. Embedded nano-micro fibrous scaffolds within hydrogel enhanced mechanical strength and physical structural anisotropy of the composite scaffold (similar to native aortic valve leaflets) and also reduced its compaction. We determined that the composite scaffolds supported the growth of human aortic valve interstitial cells (HAVIC), balanced the remodeling of heart valve ECM against shrinkage, and maintained better physiological fibroblastic phenotype in both normal and diseased HAVIC over single materials. These fabricated composite scaffolds enable the engineering of a living heart valve graft with improved anisotropic structure and tissue biomechanics important for maintaining valve cell phenotypes. Heart valve-related disease is an important clinical problem, with over 300,000 surgical repairs performed annually. Tissue engineering offers a promising strategy for heart valve repair and regeneration. In this study, we developed and tissue engineered living nano-micro fibrous woven fabric/hydrogel composite scaffolds by using textile technique combined with bioactive hydrogel formation. The novelty of our technique is that the composite scaffolds can mimic physical structure anisotropy and the mechanical strength of natural aortic valve leaflet. Moreover, the composite scaffolds prevented the matrix shrinkage, which is major problem that causes the failure of TEHV, and better maintained physiological fibroblastic phenotype in both normal and diseased HAVIC. This work marks the first report of a combination composite scaffold using 3D hydrogel enhanced by nano-micro fibrous woven fabric, and represents a promising tissue engineering strategy to treat heart valve injury.


Solution hybrid selection capture for the recovery of functional full-length eukaryotic cDNAs from complex environmental samples.
Eukaryotic microbial communities play key functional roles in soil biology and potentially represent a rich source of natural products including biocatalysts. Culture-independent molecular methods are powerful tools to isolate functional genes from uncultured microorganisms. However, none of the methods used in environmental genomics allow for a rapid isolation of numerous functional genes from eukaryotic microbial communities. We developed an original adaptation of the solution hybrid selection (SHS) for an efficient recovery of functional complementary DNAs (cDNAs) synthesized from soil-extracted polyadenylated mRNAs. This protocol was tested on the Glycoside Hydrolase 11 gene family encoding endo-xylanases for which we designed 35 explorative 31-mers capture probes. SHS was implemented on four soil eukaryotic cDNA pools. After two successive rounds of capture, >90% of the resulting cDNAs were GH11 sequences, of which 70% (38 among 53 sequenced genes) were full length. Between 1.5 and 25% of the cloned captured sequences were expressed in Saccharomyces cerevisiae. Sequencing of polymerase chain reaction-amplified GH11 gene fragments from the captured sequences highlighted hundreds of phylogenetically diverse sequences that were not yet described, in public databases. This protocol offers the possibility of performing exhaustive exploration of eukaryotic gene families within microbial communities thriving in any type of environment.


Multi-Vessel Disease in Metabolically Healthy Obese Patients Presenting with ST-Elevation Myocardial Infarction.
The extent and impact of obesity as an isolated risk factor for coronary artery disease is not clear since co-morbidities serve as confounders and may mask this association. To examine whether obesity is associated with extensive coronary artery disease among metabolically healthy patients presenting with ST-elevation myocardial infarction (STEMI) and to explore the outcomes according to body mass index (BMI). We stratified STEMI patients who had a metabolically healthy phenotype and available weight and height data according to BMI: 18.5-25 kg/m² (lean), 25.01-30 kg/m² (overweight), and > 30 kg/m² (obese). Overall 381 patients were included, 42% lean, 41% overweight, and 17% obese. Patients with increased BMIs had higher levels of low-density proteins and triglycerides (P < 0.05). Obese patients presented with the lowest rates of multi-vessel disease (12.9% vs. 22.9% for overweight and 28% for lean). In a univariable analysis, obese patients were 60% less likely to be diagnosed with multi-vessel disease (odds ratio 0.4, 95% confidence interval 0.2-0.9, P = 0.021) compared to lean patients. The association remained significant in a multivariable model adjusted for baseline characteristics (P = 0.029). There were no differences in 30-day or long-term mortality (median follow-up 3.2 years) among the groups (P > 0.1 for all comparisons). Metabolically healthy phenotype obesity was associated with lower rates of multi-vessel disease despite higher levels of triglycerides. However, this association did not translate into increased mortality.


A 19F magnetic resonance imaging-based diagnostic test for bile acid diarrhea.
In up to 50% of people diagnosed with a common ailment, diarrhea-predominant irritable bowel syndrome, diarrhea results from excess spillage of bile acids into the colon-data emerging over the past decade identified deficient release of a gut hormone, fibroblast growth factor 19 (FGF19), and a consequent lack of feedback suppression of bile acid synthesis as the most common cause. 75Selenium homotaurocholic acid (SeHCAT) testing, considered the most sensitive and specific means of identifying individuals with bile acid diarrhea, is unavailable in many countries, including the United States. Other than SeHCAT, tests to diagnose bile acid diarrhea are cumbersome, non-specific, or insufficiently validated; clinicians commonly rely on a therapeutic trial of bile acid binders. Here, we review bile acid synthesis and transport, the pathogenesis of bile acid diarrhea, the reasons clinicians frequently overlook this disorder, including the limitations of currently available tests, and our efforts to develop a novel 19F magnetic resonance imaging (MRI)-based diagnostic approach. We created 19F-labeled bile acid analogues whose in vitro and in vivo transport mimics that of naturally occurring bile acids. Using dual 1H/19F MRI of the gallbladders of live mice fed 19F-labeled bile acid analogues, we were able to differentiate wild-type mice from strains deficient in intestinal expression of a key bile acid transporter, the apical sodium-dependent bile acid transporter (ASBT), or FGF15, the mouse homologue of FGF19. In addition to reviewing our development of 19F-labeled bile acid analogue-MRI to diagnose bile acid diarrhea, we discuss challenges to its clinical implementation. A major limitation is the paucity of clinical MRI facilities equipped with the appropriate coil and software needed to detect 19F signals.


Metal accumulation in tobacco expressing Arabidopsis halleri metal hyperaccumulation gene depends on external supply.
Engineering enhanced transport of zinc to the aerial parts of plants is a major goal in bio-fortification. In Arabidopsis halleri, high constitutive expression of the AhHMA4 gene encoding a metal pump of the P(1B)-ATPase family is necessary for both Zn hyperaccumulation and the full extent of Zn and Cd hypertolerance that are characteristic of this species. In this study, an AhHMA4 cDNA was introduced into N. tabacum var. Xanthi for expression under the control of its endogenous A. halleri promoter known to confer high and cell-type specific expression levels in both A. halleri and the non-hyperaccumulator A. thaliana. The transgene was expressed at similar levels in both roots and shoots upon long-term exposure to low Zn, control, and increased Zn concentrations. A down-regulation of AhHMA4 transcript levels was detected with 10 muM Zn resupply to tobacco plants cultivated in low Zn concentrations. In general, a transcriptional regulation of AhHMA4 in tobacco contrasted with the constitutively high expression previously observed in A. halleri. Differences in root/shoot partitioning of Zn and Cd between transgenic lines and the wild type were strongly dependent on metal concentrations in the hydroponic medium. Under low Zn conditions, an increased Zn accumulation in the upper leaves in the AhHMA4-expressing lines was detected. Moreover, transgenic plants exposed to cadmium accumulated less metal than the wild type. Both modifications of zinc and cadmium accumulation are noteworthy outcomes from the biofortification perspective and healthy food production. Expression of AhHMA4 may be useful in crops grown on soils poor in Zn.


The 5-HT7 receptor as a potential target for treating drug and alcohol abuse.
Alcohol and drug abuse take a large toll on society and affected individuals. However, very few effective treatments are currently available to treat alcohol and drug addiction. Basic and clinical research has begun to provide some insights into the underlying neurobiological systems involved in the addiction process. Several neurotransmitter pathways have been implicated and distinct reward neurocircuitry have been proposed-including the mesocorticolimbic dopamine (MCL-DA) system and the extended amygdala. The serotonin (5-HT) neurotransmitter system is of particular interest and multiple 5-HT receptors are thought to play significant roles in alcohol and drug self-administration and the development of drug dependence. Among the 5-HT receptors, the 5-HT7 receptor is currently undergoing characterization as a potential target for the treatment of several psychiatric disorders. Although this receptor has received only limited research regarding addictive behaviors, aspects of its neuroanatomical, biochemical, physiological, pharmacological, and behavioral profiles suggest that it could play a key role in the addiction process. For instance, genomic studies in humans have suggested a link between variants in the gene encoding the 5-HT7 receptor and alcoholism. Recent behavioral testing using high-affinity antagonists in mice and preliminary tests with alcohol-preferring rats suggest that this receptor could mediate alcohol consumption and/or reinforcement and play a role in seeking/craving behavior. Interest in the development of new and more selective pharmacological agents for this receptor will aid in examining the 5-HT7 receptor as a novel target for treating addiction.


[Direction-changing spontaneous nystagmus with simultaneous change in pupil diameter and eyelid interspace. Oculopupillomotor syndrome in blindness due to dysgenesis mesodermalis iridis et corneae Rieger].
Case report on a 47-year-old man who got blind during the last 10 years because of dysgenesis mesodermalis iridis et corneae Rieger. The patient showed an oculopupillomotor syndrome consisting of direction-changing spontaneous nystagmus and simultaneous changes of pupil diameter and eyelid interspace. The specific modification of direction, form, frequency, and amplitude of nystagmus under central influences such as emotional excitement, change of gaze, eye closure, head tilt, and other vestibular stimuli was registered electronystagmographically and is described in this paper. The unique oculomotor disturbance is compared pathogenetically with periodic oculomotor phenomena such as periodic alternating nystagmus, other types of direction-changing nystagmus, and uninhibited oculomotoricity of the blind. Loss of cerebellar inhibition mechanisms in the central pathogenesis of this syndrome is discussed hypothetically.


Short- and long-range effects of Sonic hedgehog in limb development.
The secreted protein Sonic hedgehog (Shh) and its transmembrane receptor Patched (Ptc) control a major signal transduction pathway in early vertebrate limb development. Ligand-free Ptc interacts with the transmembrane protein Smoothened (Smo) and blocks expression of Smo-controlled genes including ptc. Ligand-bound Ptc removes the block and leads to further expression of ptc, which in turn restricts the range of Shh transport. Currently it is not certain that Shh functions as a morphogen on the 300-microm scale of early chick limb development, because it has been difficult to determine how far different forms of Shh are transported. We develop a model to study the effects of two forms of Shh used experimentally and propose a mechanism for Shh signal transduction based on a two-state model for the Ptc-Smo interaction. Recent bead- and tissue-implant experiments can be explained by using this model without postulating different diffusivities for the two forms of Shh; a difference in other parameters such as the rate of release of Shh from the bead or transplant can explain the results equally well. The model also predicts that lower concentrations of Shh in a bead will produce a response similar to that after a tissue transplant. Our results provide an explanation for the counterintuitive experimental results and show that the same signal transduction mechanism can explain both short- and long-range Shh signaling. We conclude that Shh can function as a long-range morphogen.


Identification of proteins with different abundance associated with cell migration and proliferation in leiomyoma interstitial fluid by proteomics.
Uterine leiomyoma is the most common female reproductive tract benign tumor. Little is known about protein composition and changes in the leiomyoma interstitial fluid (IF). The present study focused on changes in protein abundance in the IF of leiomyoma. Leiomyoma IFs and adjacent myometrial IFs were obtained and analyzed by two-dimensional electrophoresis (2-DE) coupled with mass spectrometry and western blotting for 2-DE data validation. A total of 25 unique proteins were observed to change significantly (P<0.05). Of these proteins with different abundance, 22 had not been previously identified in leiomyoma IF. In silico analysis predicted that three of these proteins were secreted via classical mechanisms, while 22 were secreted via non-classical mechanisms. Ingenuity Pathway Analysis identified 17 proteins associated with cellular migration and proliferation. Among these, phosphoglycerate mutase 1 had not been previously associated with leiomyoma. The abundance of seven proteins was further validated by western blotting. A comparative proteomic approach identified a number of proteins associated with cellular migration and proliferation, with changes in abundance in IF likely to be involved in tumor development. Further studies will be required to investigate the role of these proteins in leiomyoma IF and their possible association with tumor development and growth.


C-terminal domain of the mitotic apparatus protein p62 targets the protein to the nucleolus during interphase.
Mitotic apparatuses from sea urchin embryos contain a protein (p62), previously shown to be required for mitotic progression. This protein localizes to the mitotic apparatus during cell division in urchin embryos and mammalian tissue culture cells. We show here by immunofluorescence that p62 is localized to the nucleus of mammalian cells during interphase and is highly concentrated in nucleoli. In addition, a fusion protein composed of full-length p62 and green fluorescent protein also localizes to nucleoli when expressed in COS-7 cells in culture. Analysis of the primary sequence of p62 reveals three distinct domains of the protein based on amino acid charge distribution: the acidic N-terminal domain, the basic C-terminal domain, and the central, M-domain, which contains alternating subdomains of clusters of acidic and basic residues. To identify the domain important for nucleolar localization during interphase, specific domains of p62 alone, or in combination with each other or with beta-galactosidase were fused to green fluorescent protein. Following confirmation of the fusion constructs by sequence analysis, the constructs were expressed in mammalian cells, expression was confirmed by immunoblotting, and the fusion proteins were localized via fluorescence microscopy. The data demonstrate that the C-terminal domain of p62 is both necessary and sufficient for the nuclear localization and nucleolar binding of p62 that is observed during interphase.


Bench-top validation testing of selected immunological and molecular Renibacterium salmoninarum diagnostic assays by comparison with quantitative bacteriological culture.
No gold standard assay exhibiting error-free classification of results has been identified for detection of Renibacterium salmoninarum, the causative agent of salmonid bacterial kidney disease. Validation of diagnostic assays for R. salmoninarum has been hindered by its unique characteristics and biology, and difficulties in locating suitable populations of reference test animals. Infection status of fish in test populations is often unknown, and it is commonly assumed that the assay yielding the most positive results has the highest diagnostic accuracy, without consideration of misclassification of results. In this research, quantification of R. salmoninarum in samples by bacteriological culture provided a standardized measure of viable bacteria to evaluate analytical performance characteristics (sensitivity, specificity and repeatability) of non-culture assays in three matrices (phosphate-buffered saline, ovarian fluid and kidney tissue). Non-culture assays included polyclonal enzyme-linked immunosorbent assay (ELISA), direct smear fluorescent antibody technique (FAT), membrane-filtration FAT, nested polymerase chain reaction (nested PCR) and three real-time quantitative PCR assays. Injection challenge of specific pathogen-free Chinook salmon, Oncorhynchus tshawytscha (Walbaum), with R. salmoninarum was used to estimate diagnostic sensitivity and specificity. Results did not identify a single assay demonstrating the highest analytical and diagnostic performance characteristics, but revealed strengths and weaknesses of each test.


The impact of marital withdrawal and secure base script knowledge on mothers' and fathers' parenting.
The current study examines associations between marital conflict and negative parenting behaviors among fathers and mothers, and the extent to which internal working models (IWMs) of attachment relationships may serve as sources of risk or resilience during family interactions. The sample consisted of 115 families (mothers, fathers, and their 6-month-old infants) who participated in a controlled experiment. Couples were randomly assigned to engage in either a conflict or positive marital discussion, followed by parent-infant freeplay sessions and assessment of parental IWMs of attachment (i.e., secure base script knowledge). While no differences in parenting behaviors emerged between the conflict and positive groups, findings revealed that couple withdrawal during the marital discussion was related to more intrusive and emotionally disengaged parenting for mothers and fathers. Interestingly, secure base script knowledge was inversely related to intrusion and emotional disengagement for fathers, but not for mothers. Furthermore, only among fathers did secure base script knowledge serve to significantly buffer the impact of marital disengagement on negative parenting (emotional disengagement). Findings are discussed using a family systems framework and expand our understanding of families, and family members, at risk. (PsycINFO Database Record


Angiotensin-(1-7) administration reduces oxidative stress in diabetic bone marrow.
Diabetics have an increased risk of developing cardiovascular disease, in part due to oxidative stress, resulting in endothelial nitric oxide synthase (eNOS) dysfunction. Studies have demonstrated that angiotensin-(1-7) [Ang-(1-7)] can activate eNOS activity. Because the bone marrow is a primary source of a number of progenitors important in physiological homeostasis and healing, the goal of this study was to evaluate the in vivo effects of Ang-(1-7) treatment on oxidative stress and the ensuing nitrative stress in diabetic bone marrow and its potential pathways. BKS.Cg-Dock7(m) +/+ Lepr(db)/J mice and their heterozygous controls were administered Ang-(1-7) alone or combined with A-779, losartan, PD123,319, nitro-l-arginine methyl ester, or icatibant sc for 14 d. The bone marrow was then collected to measure nitric oxide levels, eNOS phosphorylation, and expression of nitric oxide synthase, superoxide dismutase, and p22-phox. Nitric oxide levels in the bone marrow were significantly decreased in diabetic mice, and Ang-(1-7) treatment was able to significantly increase these measures (P < 0.01). This effect was blocked by the coadministration of PD123,319, A-779, nitro-l-arginine methyl ester, and icatibant. In addition, Ang-(1-7) treatment reversed the paradoxical increase in eNOS and neuronal nitric oxide synthase expression and decreased the phosphorylation of eNOS at Thr495 seen in diabetic mice. Ang-(1-7) also reversed diabetes-induced production of reactive oxygen species by decreasing p22-phox expression and increasing superoxide dismutase 3 expression, leading to a significant reduction in 3-nitrotyrosine formation in diabetic bone marrow (P < 0.05). Our findings demonstrate that Ang-(1-7) administration decreases diabetes-induced oxidative stress in the bone marrow and modifies pathways involved in eNOS dysfunction.


Evaluation of the thoracolumbar injury classification system in thoracic and lumbar spinal trauma.
Retrospective study. Evaluate the relationship among the neurologic status, the Thoracolumbar Injury Classification System (TLICS) score, and the Magerl/AO classification system. A wide range of classification schemes for thoracic and lumbar spine trauma have been described, but none has achieved widespread acceptance. A recent system proposed by Vaccaro et al has been developed to improve injury classification and guide surgical decision making. Analysis of 49 patients treated surgically for thoracic and lumbar spine trauma from 2003 to 2009 in 2 spine trauma centers. Clinical and radiologic data were evaluated, classifying the trauma according to American Spinal Injury Association status, the Magerl/AO classification for fractures, and the TLICS score. The mean age was 37 years (range, 17-72). Thirty-five (71%) patients had a thoracolumbar fracture (T11-L2). A posterior approach was used in all the cases. American Spinal Injury Association status remained unchanged in 44 (4 had some improvement and 1 worsened). A total of 61.1% of the patients with a type A fracture were neurologically intact compared with 80% with complete neurologic deficit for type C fractures. The TLICS score range from 2 to 9 (average of 6.2). Forty-seven of 49 (96%) patients had a TLICS score greater than 4, suggesting surgical treatment. Seventy percentage of the patients with a TLICS score from 4 to 6 were neurologically intact compared with 87.5% of complete neurologic deficits in patients with TLICS 7 to 9. A statistic correlation was established between the neurologic status and AO type fracture (P = 0.0041) and the TLICS score (P < 0.0001). An association between the AO type fracture and the TLICS score was also found (P = 0.0088). The TLICS score treatment recommendation matched surgical treatment in 47 of 49 patients (96%). The TLICS was found to correlate to the AO classification. This suggests that the TLICS can be used to classify thoracolumbar trauma and can accurately predict surgical management.


Robust regression of enzyme kinetic data.
A method described previously [Cornish-Bowden & Endrenyi (1981) Biochem. J. 193, 1005-1008] for fitting theoretical equations to enzyme kinetic data without prior knowledge of weights or error distribution has been tested by computer simulation. With the equations for various kinds of linear inhibition as an example, the method performed well under all of the conditions examined, giving results that were often much better than those given by widely used least-squares alternatives, and were never appreciably worse. Although equations for two-substrate kinetics were not explicitly tested, the results for inhibition equations can be generalized to include two-substrate equations because the two are formally equivalent for simulation purposes. As a check on the results with inhibition equations the method was also tested for fitting bell-shaped pH-activity profiles and gave correspondingly good results.


Reliability and validity of Turkish version of pediatric balance scale.
Assessment tool is a key point to evaluate balance disturbances in children. The aim of this study was to analyze reliability and validity of Turkish version of pediatric balance scale. The present study included 34 children (13 girls; 21 boys) with balance impairments. The scale consists of 14 items. Eight observers assessed the video records of the participants for interrater agreement. One observer evaluated the records twice in 2 weeks for intrarater agreement. Intraclass correlation coefficient was used for the interobserver and intraobserver agreement. The Functional Reach Test was used to calculate concurrent validity. Mean age of the sample was 11.68 ± 3.53 years. The Turkish version of the scale was found to be reliable, perfectly (intraclass correlation coefficient for interobserver agreement = 0.915). Intraobserver agreement was also reliable, perfectly (ICC = 0.927). The strong correlation between pediatric balance scale and functional reach test was found (r = 0.692; p < 0.001). The Turkish version of the scale is a valid and reliable tool to evaluate children with balance impairments. Moreover, it is easy to use for health providers working with disabled children.


[Protracted psychoses in juvenile psychopath-like heboid schizophrenia (atypical variant of its course)].
A group of 22 patients selected as a result of follow-up examinations of 104 patients with juvenile psychopathlike heboid schizophrenia is described. A specific feature of the disease course in the patients observed consisted in appearance of atypical protracted psychoses after a long-time period of psychopathlike heboid disorders. Peculiarities differentiating this form from the slow-progressive variant of psychopathlike heboid schizophrenia and from paroxysmal schizophrenia with heboid initial stage are presented. The group described occupies an intermediate place between these forms of schizophrenia. A question on the role of aggravation of the disease course with various exogenous adverse factors (noted in all the cases described) is discussed.


Lopinavir/ritonavir: a review of its use in the management of HIV infection.
Lopinavir is a novel protease inhibitor (PI) developed from ritonavir. Coadministration with low-dose ritonavir significantly improves the pharmacokinetic properties and hence the activity of lopinavir against HIV-1 protease. Coformulated lopinavir/ritonavir was developed for ease of administration and to ensure both drugs are taken together, as part of combination therapy with other antiretroviral agents. Coformulated lopinavir/ritonavir-based regimens provide adequate and durable suppression of viral load and sustained improvements in CD4+ cell counts, as demonstrated in randomised trials in antiretroviral therapy-naive and -experienced adults and children. To date, development of primary resistance to lopinavir/ritonavir has not been observed in 470 antiretroviral therapy-naive patients treated for >48 weeks. The lopinavir/ritonavir-based regimen was more effective than nelfinavir in antiretroviral therapy-naive HIV-1-infected patients in a phase III trial. The coformulation is also effective as 'salvage' therapy, as shown by low cross-resistance rates in patients who failed to respond to treatment with other PIs in phase II trials. Coformulated lopinavir/ritonavir was well tolerated in both antiretroviral therapy-naive and -experienced HIV-1-infected adults and children with low rates of study drug-related treatment discontinuations. The most common adverse event in adults associated with lopinavir/ritonavir was diarrhoea, followed by other gastrointestinal disturbances, asthenia, headache and skin rash. The incidence of moderate-to-severe adverse events in children was low, skin rash being the most common. Changes in body fat composition occurred with equal frequency in lopinavir/ritonavir- and nelfinavir-treated naive patients, through week 60 in a phase III study. Although laboratory abnormalities occurred with similar frequency in both treatment groups, triglycerides grade 3/4 elevations were significantly more frequent with lopinavir/ritonavir. Total cholesterol and triglycerides grade 3/4 elevations appear to occur more frequently in PI-experienced than in PI-naive lopinavir/ritonavir-treated patients. A number of clinically important drug interactions have been reported with lopinavir/ritonavir necessitating dosage adjustments of lopinavir/ritonavir and/or the interacting drugs, and several other drugs are contraindicated in patients receiving the coformulation. Coformulated lopinavir/ritonavir is a novel PI that, in combination with other antiretroviral agents, suppresses plasma viral load and enhances immunological status in therapy-naive and -experienced patients with HIV-1 infection. Lopinavir/ritonavir appears more effective than nelfinavir in 'naive' patients and is also suitable for 'salvage' therapy, because of its high barrier to development of resistance. Given its clinical efficacy, a tolerability profile in keeping with this class of drugs, favourable resistance profile and easy-to-adhere-to administration regimen, coformulated lopinavir/ritonavir should be regarded as a first-line option when including a PI in the management of HIV-1 infection. Lopinavir/ritonavir is a coformulation of two structurally related protease inhibitor (PI) antiretroviral agents. Lopinavir is a highly potent and selective inhibitor of the HIV type 1 (HIV-1) protease, an essential enzyme for production of mature, infective virus. It acts by arresting maturation of HIV-1 thereby blocking its infectivity. Thus, the main antiviral action of lopinavir is to prevent subsequent infections of susceptible cells; it has no effect on cells with already integrated viral DNA. Lopinavir has an approximate, equals 10-fold higher in vitro activity against both wild-type and mutant HIV-1 proteases than ritonavir; however, its in vivo activity is greatly attenuated by a high first-pass hepatic metabolism. The low-dose ritonavir coadministered with lopinavir inhibits metabolic inactivation of lopinavir and acts only as its pharmacokinetic enhancer. Therefore, the antiretroviral activity of roviral activity of coformulated lopinavir/ritonavir 400/100mg twice daily is derived solely from lopinavir plasma concentrations. Combining lopinavir with low-dose ritonavir produces lopinavir concentrations far exceeding those needed to suppress 50% of in vitro and in vivo viral replication in CD4+ cells and monocyte/macrophages (main human reservoirs of HIV-1 infection). Thus far, no resistance to lopinavir has been detected in clinical trials in antiretroviral therapy-naive patients treated for up to 204 weeks and only 12% of HIV-1 strains from patients in whom prior treatment with multiple PIs have failed, have been observed to develop resistance to coformulated lopinavir/ritonavir. A strong negative correlation was found between the number of PI mutations at baseline and the viral response rates achieved with lopinavir/ritonavir-based regimens in PI-experienced patients, indicating that resistance to lopinavir increases with increasing number of PI mutations and that five PI mutations represent the clinically relevant genotypic breakpoint for lopinavir. The absolute bioavailability of lopinavir coformulated with ritonavir in humans has not yet been established. Multiple-dosage absorption pharmacokinetics of lopinavir/ritonavir 400/100mg twice daily (the mean peak [C(max)] and trough [C(trough)] plasma concentrations at steady-state and the 12-hour area under the plasma concentration-time curve [AUC(12)] of either drug) were stable in antiretroviral therapy-naive and single PI-experienced adult patients receiving therapy over a 24-week evaluation period. The C(trough) values of lopinavir, achieved with lopinavir/ritonavir 400/100mg twice daily, were median 84-fold higher than the protein binding-adjusted 50% effective concentration (EC(50)) of lopinavir against wild-type HIV-1 in antiretroviral therapy-naive HIV-1-infected patients in a phase II study. Bioavailability of lopinavir administered in either the capsule or the liquid lopinavir/ritonavir formulation can be increased substantially with concurrent ingestion of food with moderate-to-high fat content. At steady state, lopinavir is approximately 98-99% plasma protein bound and the percentage of its unbound (i.e. pharmacologically active) fraction is dependent on total drug plasma concentration. Both lopinavir and ritonavir penetrate poorly into the human genital tracts and the cerebrospinal fluid. Both agents undergo extensive and rapid first-pass metabolism by hepatic cytochrome P450 (CYP) 3A4 isoenzyme. However, ritonavir also potently inhibits this enzyme and acts as a pharmacokinetic enhancer of lopinavir. The elimination half-life and apparent oral clearance of lopinavir average approximately 4-6 hours and approximately 6-7 L/h, respectively, with lopinavir/ritonavir 400/100mg twice daily administration. Less than 3% and 20% of the lopinavir dose is excreted unchanged in the urine and faeces, respectively. Limited data show similar pharmacokinetics of lopinavir in children as in adults. Coformulated lopinavir/ritonavir has the potential to interact with wide variety of drugs via several mechanisms, mostly involving the CYP enzymes. Coadministration of lopinavir/ritonavir is contraindicated with certain drugs (i.e. flecainide, propafenone, astemizole, terfenadine, ergot derivatives, cisapride, pimozide, midazolam and triazolam) that are highly dependent on CYP3A or CYP2D6 for clearance and for which elevated plasma concentrations are associated with serious and/or life-threatening events. Coadministration with lopinavir/ritonavir is also not recommended for drugs or herbal products (i.e. rifampicin [rifampin] and St. John's wort [Hypericum perforatum]) that may substantially reduce lopinavir plasma concentrations, or drugs whose plasma concentrations elevated by the coformulation may lead to serious adverse reactions (i.e. simvastatin and lovastatin). However, a recent study in healthy volunteers suggests that adequate lopinavir concentrations may be achieved during rifampicin coadministration by increasing the twice-daily dosage of lopinavir/ritonavir in conjunction with therapeutic drug monitoring. The liquid (but not the capsule) formulation of lopinavir/ritonavir contains 42.4% ethanol (v/v) and should not be coadministered with drugs capable of producing disulfiram-like reactions (e.g. disulfiram, metronidazole). Coadministration with saquinavir or indinavir requires no dosage adjustment, whereas coadministration with amprenavir, nevirapine or efavirenz requires a dosage increase of the coformulation typically by 33%. As the oral bioavailability of both didanosine and lopinavir/ritonavir is significantly affected by concurrent food ingestion, didanosine should be administered 1 hour before or 2 hours after lopinavir/ritonavir has been taken with food. Interactions between lopinavir/ritonavir and other nucleoside reverse transcriptase inhibitors (NRTIs) are not expected. The coformulation is also likely to increase plasma concentrations of non-antiretroviral drugs metabolised through the CYP3A pathway. To reduce the risk of their toxicity when coadministered with lopinavir/ritonavir, the recommended actions include: (i) monitoring of the drug plasma concentration (antiarrhythmics and immunosuppressants) or the international normalised ratio (warfarin); (ii) the use of alternative treatment (atorvastatin) or birth control methods (ethinylestradiol); and (iii) dosage adjustment (clarithromycin [only in patients with renal failure], rifabutin, dihydropyridine calcium-channel blockers, atorvastatin, ketoconazole and itraconazole). (ABSTRACT TRUNCATED)


Critical role of the C5a-activated neutrophils in high-fat diet-induced vascular inflammation.
Exceed and chronic high-fat diet (HFD) contributes to the diagnosis and development of atherosclerosis, obesity, and metabolic syndrome. However, the key molecular component(s) triggered by HFD responsible for initiating vascular inflammation remain unknown. We observed that feeding HFD for 4 weeks is sufficient to induce leukocyte recruitment in the femoral artery of wild-type mice. Neutrophil- and monocyte-depletion analyses confirmed the preferential recruitment of neutrophils in these mice. Protein analysis of sera from HFD-fed mice revealed a marked elevation of complement component C5a levels. Exogenous C5a alone induced leukocyte recruitment, which was abolished by a C5a-receptor antagonist. We also examined the role of neutrophil-derived MCP-1 in accumulation of leukocytes in the artery. These results demonstrated a previously unrecognized role for C5a and neutrophils in the early onset of HFD-induced vascular inflammation. Further study may help in elucidating a novel regulatory pathway to control diet-induced inflammation such as that in case of atherosclerosis.


Altered purinergic signaling in uridine adenosine tetraphosphate-induced coronary relaxation in swine with metabolic derangement.
We previously demonstrated that uridine adenosine tetraphosphate (Up4A) induces potent and partially endothelium-dependent relaxation in the healthy porcine coronary microvasculature. We subsequently showed that Up4A-induced porcine coronary relaxation was impaired via downregulation of P1 receptors after myocardial infarction. In view of the deleterious effect of metabolic derangement on vascular function, we hypothesized that the coronary vasodilator response to Up4A is impaired in metabolic derangement, and that the involvement of purinergic receptor subtypes and endothelium-derived vasoactive factors (EDVFs) is altered. Coronary small arteries, dissected from the apex of healthy swine and swine 6 months after induction of diabetes with streptozotocin and fed a high-fat diet, were mounted on wire myographs. Up4A (10-9-10-5 M)-induced coronary relaxation was maintained in swine with metabolic derangement compared to normal swine, despite impaired endothelium-dependent relaxation to bradykinin and despite blunted P2X7 receptor and NO-mediated vasodilator influences of Up4A. Moreover, a thromboxane-mediated vasoconstrictor influence was unmasked. In contrast, an increased Up4A-mediated vasodilator influence via P2Y1 receptors was observed, while, in response to Up4A, cytochrome P450 2C9 switched from producing vasoconstrictor to vasodilator metabolites in swine with metabolic derangement. Coronary vascular expression of A2A and P2X7 receptors as well as eNOS, as assessed with real-time PCR, was reduced in swine with metabolic derangement. In conclusion, although the overall coronary vasodilator response to Up4A was maintained in swine with metabolic derangement, the involvement of purinergic receptor subtypes and EDVF was markedly altered, revealing compensatory mechanisms among signaling pathways in Up4A-mediated coronary vasomotor influence in the early phase of metabolic derangement. Future studies are warranted to investigate the effects of severe metabolic derangement on coronary responses to Up4A.


Diagnosis of Choroidal Disease With Deep Learning-Based Image Enhancement and Volumetric Quantification of Optical Coherence Tomography.
The purpose of this study was to quantify choroidal vessels (CVs) in pathological eyes in three dimensions (3D) using optical coherence tomography (OCT) and a deep-learning analysis. A single-center retrospective study including 34 eyes of 34 patients (7 women and 27 men) with treatment-naïve central serous chorioretinopathy (CSC) and 33 eyes of 17 patients (7 women and 10 men) with Vogt-Koyanagi-Harada disease (VKH) or sympathetic ophthalmitis (SO) were imaged consecutively between October 2012 and May 2019 with a swept source OCT. Seventy-seven eyes of 39 age-matched volunteers (26 women and 13 men) with no sign of ocular pathology were imaged for comparison. Deep-learning-based image enhancement pipeline enabled CV segmentation and visualization in 3D, after which quantitative vessel volume maps were acquired to compare normal and diseased eyes and to track the clinical course of eyes in the disease group. Region-based vessel volumes and vessel indices were utilized for disease diagnosis. OCT-based CV volume maps disclose regional CV changes in patients with CSC, VKH, or SO. Three metrics, (i) choroidal volume, (ii) CV volume, and (iii) CV index, exhibit high sensitivity and specificity in discriminating pathological choroids from healthy ones. The deep-learning analysis of OCT images described here provides a 3D visualization of the choroid, and allows quantification of features in the datasets to identify choroidal disease and distinguish between different diseases. This novel analysis can be applied retrospectively to existing OCT datasets, and it represents a significant advance toward the automated diagnosis of choroidal pathologies based on observations and quantifications of the vasculature.


Lycopene ameliorates PTSD-like behaviors in mice and rebalances the neuroinflammatory response and oxidative stress in the brain.
Posttraumatic stress disorder (PTSD) is a mental disorder that can translate into severe economic problems. Lycopene is an aliphatic hydrocarbon carotenoid extracted from plants, including papayas, tomatoes, and water melons. Previous studies have shown that lycopene can produce antidepressant-like effects in rodent models of depression. However, little is known about its anti-PTSD-like effect. This was addressed in the present study by using the single prolonged stress (SPS) protocol to induce PTSD-like behavioral deficits in mice. Our results showed that 12 days of lycopene treatment at the dose of 10 and 20 mg/kg, but not at 5 mg/kg, ameliorated the PTSD-like phenotype induced by SPS, including the increase in freezing time in contextual fear paradigm, the decrease in time and entries in open arms in elevated plus maze test, and the decrease in distance and time in the central area of the open field test, without affecting the mouse locomotor activity. Mechanistic studies revealed that lycopene treatment (20 mg/kg, 12 days) could suppress the SPS-induced increase in levels of interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α), and nitrite in the hippocampus and prefrontal cortex in mice, as well as the increased markers that indicate high levels of oxido-nitrosative stress in the hippocampus and prefrontal cortex in SPS mice. Lycopene treatment (20 mg/kg, 12 days) also suppressed the SPS-induced decrease in brain derived neurotrophic factor (BDNF) levels in the hippocampus and prefrontal cortex in mice. Overall, the anti-PTSD-like effect of lycopene may be associated with its anti-neuroinflammation and anti-oxidative stress activities.


Behavioral Communication Strategies for Global Epidemics: An Innovative Model for Public Health Education and Humanitarian Response.
In response to a number of growing global health challenges, New York University and UNICEF designed a Behavioral Communication Strategies for Global Epidemics course that brings together United Nations professionals, government staff, and MPH (Master of Public Health) students to design innovative social behavior change communication (SBCC) strategies that address disease outbreaks and humanitarian challenges around the world. Applying a systems approach, participants in the course work on interdisciplinary teams to design strategies, develop skills, and engage in global learning. At the culmination of the course, all teams present strategies to UNICEF country offices for implementation. This innovative model for disease outbreak, public health education, and humanitarian response provides professionals with an opportunity to develop a wide range of competencies, including systems thinking, behavior change, and human-centered design and equips them with the necessary tools to develop more novel approaches to SBCC. As the number of outbreaks and humanitarian challenges increase each year, this format for learning can serve as a model for how professionals can effectively address these complex crises.


[Modes of mechanical ventilation during transferring the patient to spontaneous breathing].
Mechanical ventilation (MV) has become a general treatment in the intensive care unit in recent years. Mechanical ventilation is a resuscitation treatment; however MV causes many implications therefore it is to be finished as soon as the patient's condition begins improve. Modern transferring the patient to spontaneous breathing decreases implications number. Significant part of mechanical ventilation time (40%) is a time of weaning from mechanical ventilation. Weaning from MV is an economical, clinical and ethical problem. Many ventilation modes have introduced in clinical practice through the microprocessor technologies development. Supporting ventilation modes help to avoid some adverse effects of mechanical ventilation. The article deals with historical approaches development their advantages and limitations.


Thymus transplantation in complete DiGeorge syndrome: immunologic and safety evaluations in 12 patients.
Complete DiGeorge syndrome is a fatal condition in which infants have no detectable thymus function. The optimal treatment for the immune deficiency of complete DiGeorge syndrome has not been determined. Safety and efficacy of thymus transplantation were evaluated in 12 infants with complete DiGeorge syndrome who had less than 20-fold proliferative responses to phytohemagglutinin. All but one had fewer than 50 T cells/mm3. Allogeneic postnatal cultured thymus tissue was transplanted. T-cell development was followed by flow cytometry, lymphocyte proliferation assays, and T-cell receptor Vbeta (TCRBV) repertoire evaluation. Of the 12 patients, 7 are at home 15 months to 8.5 years after transplantation. All 7 survivors developed T-cell proliferative responses to mitogens of more than 100 000 counts per minute (cpm). By one year after transplantation, 6 of 7 patients developed antigen-specific proliferative responses. The TCRBV repertoire showed initial oligoclonality that progressed to polyclonality within a year. B-cell function developed in all 3 patients tested after 2 years. Deaths were associated with underlying congenital problems. Risk factors for death included tracheostomy, long-term mechanical ventilation, and cytomegalovirus infection. Adverse events in the first 3 months after transplantation included eosinophilia, rash, lymphadenopathy, development of CD4-CD8- peripheral T cells, elevated serum immunoglobulin E (IgE), and possible pulmonary inflammation. Adverse events related to the immune system occurring more than 3 months after transplantation included thrombocytopenia in one patient and hypothyroidism and alopecia in one other patient. Thymic transplantation is efficacious, well tolerated, and should be considered as treatment for infants with complete DiGeorge syndrome.


A novel method for simulating insulin mediated GLUT4 translocation.
Glucose transport in humans is a vital process which is tightly regulated by the endocrine system. Specifically, the insulin hormone triggers a cascade of intracellular signals in target cells mediating the uptake of glucose. Insulin signaling triggers cellular relocalization of the glucose transporter protein GLUT4 to the cell surface, which is primarily responsible for regulated glucose import. Pathology associated with the disruption of this pathway can lead to metabolic disorders, such as type II diabetes mellitus, characterized by the failure of cells to appropriately uptake glucose from the blood. We describe a novel simulation tool of the insulin intracellular response, incorporating the latest findings regarding As160 and GEF interactions. The simulation tool differs from previous computational approaches which employ algebraic or differential equations; instead, the tool incorporates statistical variations of kinetic constants and initial molecular concentrations which more accurately mimic the intracellular environment. Using this approach, we successfully recapitulate observed in vitro insulin responses, plus the effects of Wortmannin-like inhibition of the pathway. The developed tool provides insight into transient changes in molecule concentrations throughout the insulin signaling pathway, and may be employed to identify or evaluate potentially critical components of this pathway, including those associated with insulin resistance. In the future, this highly tractable platform may be useful for simulating other complex cell signaling pathways. Biotechnol. Bioeng. 2014;111: 2454-2465. © 2014 Wiley Periodicals, Inc.


Oligogalacturonic Acid Inhibits Vascular Calcification by Two Mechanisms: Inhibition of Vascular Smooth Muscle Cell Osteogenic Conversion and Interaction With Collagen.
Cardiovascular diseases constitute the leading cause of mortality worldwide. Calcification of the vessel wall is associated with cardiovascular morbidity and mortality in patients having many diseases, including diabetes mellitus, atherosclerosis, and chronic kidney disease. Vascular calcification is actively regulated by inductive and inhibitory mechanisms (including vascular smooth muscle cell adaptation) and results from an active osteogenic process. During the calcification process, extracellular vesicles (also known as matrix vesicles) released by vascular smooth muscle cells interact with type I collagen and then act as nucleating foci for calcium crystallization. Our primary objective was to identify new, natural molecules that inhibit the vascular calcification process. We have found that oligogalacturonic acids (obtained by the acid hydrolysis of polygalacturonic acid) reduce in vitro inorganic phosphate-induced calcification of vascular smooth muscle cells by 80% and inorganic phosphate-induced calcification of isolated rat aortic rings by 50%. A specific oligogalacturonic acid with a degree of polymerization of 8 (DP8) was found to inhibit the expression of osteogenic markers and, thus, prevent the conversion of vascular smooth muscle cells into osteoblast-like cells. We also evidenced in biochemical and immunofluorescence assays a direct interaction between matrix vesicles and type I collagen via the GFOGER sequence (where single letter amino acid nomenclature is used, O=hydroxyproline) thought to be involved in interactions with several pairs of integrins. DP8 inhibits vascular calcification development mainly by inhibition of osteogenic marker expression but also partly by masking the GFOGER sequence-thereby, preventing matrix vesicles from binding to type I collagen.


Meproadifen reaction with the ionic channel of the acetylcholine receptor: potentiation of agonist-induced desensitization at the frog neuromuscular junction.
The actions of the nicotinic noncompetitive antagonist meproadifen on both the acetylcholine (ACh) receptor-ion channel complex and electrically excitable membrane were examined in frog sciatic-nerve sartorius muscle preparations. Meproadifen (10-25 microM) blocked the nerve-evoked twitch without affecting the directly evoked twitch, the threshold, overshoot, amplitude, rate of rise, or falling phase of the directly elicited action potential in muscle. This suggests that this agent, at the concentrations that affect the nicotinic receptor, had negligible effect on the excitable membrane. In addition, the drug did not affect either the quantal content or quantal size of the end-plate potential. Meproadifen caused a voltage- and time-dependent decrease in the peak amplitude of the end-plate current (EPC) without significantly shortening the time constant of EPC decay. The voltage- and time-dependent effects of meproadifen were more pronounced at more negative potentials, as evidenced by hysteresis loops and nonlinearity in the current-voltage relationship of the EPC. Both hysteresis and nonlinearity in the current-voltage relationship of the EPC were eliminated when brief conditioning pulses were used for stepwise changes of membrane potentials. The decay time constant of the EPC in the presence of meproadifen remained an exponential function of time. Meproadifen blocked iontophoretically elicited EPCs but did not affect single-channel lifetime, conductance, or the decay time constant of the miniature EPC. Thus, the blockade was more marked on iontophoretically elicited EPCs than on miniature EPCs. Meproadifen also caused desensitization of both the junctional and extrajunctional ACh receptors, but, more important, meproadifen accelerated steady-state desensitization by several-fold (compared with the agonist). The marked depression of peak EPC amplitude and miniature EPC, its high affinity for the binding sites in the presence of the agonist, and acceleration of agonist-induced desensitization suggest that meproadifen interacts with the ACh-bound but nonconducting state of the ACh receptor-ion channel complex. Therefore, it appears that meproadifen interacts with the closed ionic channel of the ACh receptor in its resting and activated but nonconducting states, and only slightly affects the open conformation of the ionic channel.


DNA-directed coupling of organic modules by multiple parallel reductive aminations and subsequent cleavage of selected DNA sequences.
A new method for DNA-directed assembly of organic modules by multiple parallel reductive aminations is presented. Linear oligonucleotide-functionalized modules (LOMs) consist of a rigid oligo(phenylene ethynylene) backbone with two salicylaldehyde termini, and each terminus is conjugated with an oligonucleotide sequence. The stability of the tetrahydrosalen-linked modules toward elevated temperature, low pH, nucleophiles, and metal chelators is studied and compared to the analogous metal-salen-linked modules. A linear oligonucleotide-functionalized disulfide-linked module (LOSM) containing cleavable linkers between the organic module and the two DNA sequences is coupled by DNA-directed reductive aminations to non-modified LOM modules. This enables selective cleavage of the DNA strands of a central module in a structure consisting of three modules, and the reactions are analyzed by electrophoresis and 32P-labeling of one of the DNA sequences of the central LOSM.


Polarographic measurement of hydrogen sulfide production and consumption by mammalian tissues.
The role of nitric oxide (NO) in redox cell signaling is widely accepted. However, the biological role of another candidate small inorganic signaling molecule and the subject of this study, hydrogen sulfide (H2S), is much less known. H2S as a reductant and nucleophile has numerous potential cellular targets; however, its rapid biological oxidation suggests a fleeting cellular existence. The challenge of accurate real-time measurement of H2S at low micromolar or nanomolar concentrations in biological preparations represents a major impediment to H2S investigations. We here demonstrate the use of a novel polarographic H2S sensor (PHSS) to follow rapid changes in H2S concentration in common buffered biological solutions with a detection limit near 10 nM. The PHSS, used in combination with O2 and NO sensors in multisensor respirometry, shows stability, a high signal-to-noise ratio, and signal specificity for H2S. Preparations of rat vascular tissue exhibit H2S production on the addition of sulfhydryl-bearing amino acid substrates and H2S consumption when supplied with exogenous H2S. Taken together, these findings suggest the existence of dynamic steady-state cellular H2S levels. The PHSS should facilitate the investigation of H2S biology by providing a previously unattainable continuous record of H2S under biologically relevant conditions.


Gap theory based post-occupancy evaluation (GTbPOE) of dormitory building performance: A case study and a comparative analysis.
Student housing significantly influences the quality and competitiveness of the university education environment. Whereas the traditional post-occupancy evaluations (POEs) of buildings have typically focussed on investigating users' satisfaction, an earlier study developed the gap theory based post-occupancy evaluation (GTbPOE) method, by which both the users' expectation and satisfaction (viz. performance gap) of a university dormitory were investigated. To validate the applicability of the GTbPOE method, further research was undertaken to evaluate the building performance of another dormitory. Using face-to-face interviews, responses of 104 dormitory users were collected, of which the relative importance ratings of six essential aspects (namely: visual comfort, thermal comfort, aural comfort, fire safety, hygiene, and communication via information technology) were analyzed via the Analytic Hierarchy Process (AHP). A series of gap analyses on the users' expectation and satisfaction levels corroborated the existence of the adaptation effect on the users' perception: that is, the longer the stay, the smaller the performance gap. A comparative analysis on the findings between the two dormitories - one from the earlier study and the other from the present study - further demonstrated the usefulness of the GTbPOE method in benchmarking building performance. Adoption of this method in future POE studies will enable reliable identification of any shortcoming in building performance and hence, can form the basis for improvement measures to augment the performance of buildings within the built environment.


Phase I clinical study of AZD2171, an oral vascular endothelial growth factor signaling inhibitor, in patients with advanced solid tumors.
AZD2171 is a highly potent oral selective inhibitor of vascular endothelial growth factor (VEGF) signaling. This phase I study was designed to evaluate the safety and tolerability of increasing doses of AZD2171, with additional assessments of pharmacokinetics, pharmacodynamics, and efficacy. In part A, 36 patients with solid tumors and liver metastases refractory to standard therapies received once-daily oral AZD2171 (0.5 to 60 mg). Doses were escalated in successive cohorts until the maximum-tolerated dose was identified. In part B, patients with (n = 36) or without (n = 11) liver metastases were randomly assigned to receive once-daily AZD2171 (20, 30, or 45 mg). In both parts, treatment continued until tumor progression or dose-limiting toxicity (DLT) was observed. Eighty-three patients received AZD2171, which was generally well tolerated at doses of 45 mg/d or less; the most frequently reported dose-related adverse events were diarrhea, dysphonia, and hypertension. The most common DLT was hypertension (n = 7), which occurred at AZD2171 doses of 20 mg and higher. After a single dose, maximum plasma (peak) drug concentration after single-dose administration (Cmax) was achieved 1 to 8 hours postdosing with an arithmetic mean half-life associated with terminal slope of a semilogarithmic concentration-time curve (t1/2 lamda(z)) of 22 hours. Pharmacodynamic assessments demonstrated time-, dose-, and exposure-related decreases in initial area under the curve, defined over 60 seconds post-contrast arrival in the tissue (iAUC60) using dynamic contrast-enhanced magnetic resonance imaging, as well as dose- and time-dependent reductions in soluble VEGF receptor 2 levels. Preliminary evidence of efficacy included two confirmed partial responses and 22 patients with stable disease; effects on tumor size appeared to be dose related. Once-daily oral AZD2171 at doses of 45 mg or less was generally well tolerated and was associated with encouraging antitumor activity in patients with a broad range of advanced solid tumors.


Characterization of the type strain of Campylobacter coli, CIP 70.80, by plasmid typing.
A 1.9-kb plasmid DNA fragment from the type strain of Campylobacter coli, CIP 70.80, was used as a probe to characterize this type strain, other C. coli type strains obtained from several culture collections, and other C. coli strains. A specific hybridization pattern was obtained, and this pattern can be used to identify, characterize, and follow up C. coli type strains in culture collections.


Novel gene activated in rat insulinomas.
Insulinomas can be induced in experimental animals by the combined administration of diabetogenic agents with polyadenosine diphosphate (polyADP)-ribose synthetase inhibitors. A complementary DNA (cDNA) library that was constructed from streptozocin-nicotinamide-induced rat insulinomas has been found to contain a novel gene encoding a basic protein of 145 amino acids. The gene was expressed in alloxan-nicotinamide-induced insulinomas as well as in streptozocin-nicotinamide-induced insulinomas but not in normal pancreatic islets or in regenerating islets. This indicates that the activation of the gene designated rig, i.e., rat insulinoma gene, may be a general feature of pancreatic beta-cell transformation.


Optimization of scarless human stem cell genome editing.
Efficient strategies for precise genome editing in human-induced pluripotent cells (hiPSCs) will enable sophisticated genome engineering for research and clinical purposes. The development of programmable sequence-specific nucleases such as Transcription Activator-Like Effectors Nucleases (TALENs) and Cas9-gRNA allows genetic modifications to be made more efficiently at targeted sites of interest. However, many opportunities remain to optimize these tools and to enlarge their spheres of application. We present several improvements: First, we developed functional re-coded TALEs (reTALEs), which not only enable simple one-pot TALE synthesis but also allow TALE-based applications to be performed using lentiviral vectors. We then compared genome-editing efficiencies in hiPSCs mediated by 15 pairs of reTALENs and Cas9-gRNA targeting CCR5 and optimized ssODN design in conjunction with both methods for introducing specific mutations. We found Cas9-gRNA achieved 7-8× higher non-homologous end joining efficiencies (3%) than reTALENs (0.4%) and moderately superior homology-directed repair efficiencies (1.0 versus 0.6%) when combined with ssODN donors in hiPSCs. Using the optimal design, we demonstrated a streamlined process to generated seamlessly genome corrected hiPSCs within 3 weeks.


Acute macular neuroretinopathy: long-term insights revealed by multimodal imaging.
To report the structural and functional changes in acute macular neuroretinopathy (AMN) and their long-term evolution. Multimodal retinal imaging was acquired, including Fourier domain optical coherence tomography (OCT), infrared (IR) reflectance, and near IR autofluorescence (NIA). In this retrospective observational case series, detailed clinical history and multimodal imaging are reported in eight patients with AMN. Manual segmentation of the Fourier domain OCT volume scans was done in one patient with the largest AMN lesion to yield retinal sublayer topographic maps. Two patients were seen within the first 1 to 2 days of symptoms, and both showed outer nuclear and outer plexiform layer hyperreflectivity. Both patients developed enlargement of the lesion over the first week on IR reflectance imaging with a corresponding lateral extension of the outer retinal disruption on Fourier domain OCT. Thinning of the outer nuclear layer persisted in all patients with lesions >100 μm width, and in one patient this thinning worsened over the course of follow-up, as noted on the sublayer maps. This structural abnormality correlated with long-term functional deficits, persisting up to 14 months after the initial episode. Infrared reflectance highlights the lesion best, and abnormalities on near IR autofluorescence may be present. Acute macular neuroretinopathy acutely affects the outer nuclear and plexiform layers manifesting as OCT hyperreflectivity. The hallmark long-term changes are outer nuclear thinning on Fourier domain OCT and a fading dark lesion on IR reflectance imaging. These changes correspond to focal disruption of the outer segment/retinal pigment epithelium junction on OCT, and not the inner segment/outer segment junction, as previously reported. Optical coherence tomography and near IR autofluorescence abnormalities suggest previously unrecognized melanin and retinal pigment epithelium derangements in this condition.


Lack of reduction of thymidine kinase activity in stavudine-treated HIV-infected patients.
It has been demonstrated that prolonged in vivo or in vitro treatment with some nucleosides analogs may favor the selection of cells with a reduced activity of enzymes involved in the phosphorylation of these drugs leading to a reduced sensitivity to their antiretroviral action. The aim of this study was to evaluate the effect, in vivo, of zidovudine and stavudine treatment on thymidine kinase (TK) activity. The results obtained showed that TK levels in PBMC from naive patients and stavudine-treated patients did not significantly differ (naive TK = 4.16 +/- 1.19 U/mg protein; stavudine TK = 3.65 +/- 1.73 U/mg protein; p = 0.42), suggesting that the treatment with this nucleoside analog is not associated to a defect of TK activity. On the contrary, PBMC from zidovudine-treated patients showed a significant reduction in TK activity compared to naive patients (naive TK = 4.16 +/- 1.19 U/mg protein; zidovudine TK = 2.70 +/- 1.54; p = 0.014. Although the clinical significance of these results has to be established, we can speculate that stavudine and zidovudine, which are presumably phosphorylated by the same cellular kinases, might display a different ability to in vivo select cells with a resistant phenotype.


Clinicopathological characteristics in patients presenting with acute onset of symptoms caused by Rathke's cleft cysts.
Symptomatic Rathke's cleft cyst is usually accompanied by a long history of headache, visual disturbance, and hypopituitarism; however, rare cases present with acute onset and the clinical features in such cases remain uncertain. We report herein the clinical features of Rathke's cleft cyst with acute onset and discuss the clinical significance. In this study, we defined acute onset as the clinical course with clinical symptoms within a 7-day history. From among 35 cases of symptomatic Rathke's cleft cyst that were pathologically diagnosed at Fukuoka University Hospital between 1990 and 2009, five cases presented with acute onset. The symptoms, endocrinological findings, MR image findings, and pathological findings of these cases were analyzed retrospectively. Mean age was 56.8 years. Initial symptoms included headache (n = 3), general malaise (n = 2), polyuria (n = 2), and fever (n = 1). MR imaging revealed an intrasellar cystic lesion with suprasellar extension in all cases and showed rim enhancement in three cases. All cases were treated by transsphenoidal surgery. Pathological findings included hemorrhage (n = 2), hypophysitis (n = 2), and abscess formation in the cyst (n = 1). Postoperatively, all symptoms, except for hypopituitarism, improved in all cases. Rathke's cleft cysts sometimes present with acute onset, and the presentation is consistent with the features of pituitary apoplexy caused by pituitary adenoma. Although pituitary apoplexy due to hemorrhage, inflammation, or infection due to an underlying Rathke's cleft cyst is difficult to diagnose pre-operatively, Rathke's cleft cyst should be included in the differential diagnosis, and early surgical treatment is needed, as for pituitary apoplexy caused by pituitary adenoma.


Increased B Cell Selection Stringency In Germinal Centers Can Explain Improved COVID-19 Vaccine Efficacies With Low Dose Prime or Delayed Boost.
The efficacy of COVID-19 vaccines appears to depend in complex ways on the vaccine dosage and the interval between the prime and boost doses. Unexpectedly, lower dose prime and longer prime-boost intervals have yielded higher efficacies in clinical trials. To elucidate the origins of these effects, we developed a stochastic simulation model of the germinal center (GC) reaction and predicted the antibody responses elicited by different vaccination protocols. The simulations predicted that a lower dose prime could increase the selection stringency in GCs due to reduced antigen availability, resulting in the selection of GC B cells with higher affinities for the target antigen. The boost could relax this selection stringency and allow the expansion of the higher affinity GC B cells selected, improving the overall response. With a longer dosing interval, the decay in the antigen with time following the prime could further increase the selection stringency, amplifying this effect. The effect remained in our simulations even when new GCs following the boost had to be seeded by memory B cells formed following the prime. These predictions offer a plausible explanation of the observed paradoxical effects of dosage and dosing interval on vaccine efficacy. Tuning the selection stringency in the GCs using prime-boost dosages and dosing intervals as handles may help improve vaccine efficacies.


Identification and characterization of a highly motile and antibiotic refractory subpopulation involved in the expansion of swarming colonies of Paenibacillus vortex.
Bacteria often use sophisticated cooperative behaviours, such as the development of complex colonies, elaborate biofilms and advanced dispersal strategies, to cope with the harsh and variable conditions of natural habitats, including the presence of antibiotics. Paenibacillus vortex uses swarming motility and cell-to-cell communication to form complex, structured colonies. The modular organization of P. vortex colony has been found to facilitate its dispersal on agar surfaces. The current study reveals that the complex structure of the colony is generated by the coexistence and transition between two morphotypes--'builders' and 'explorers'--with distinct functions in colony formation. Here, we focused on the explorers, which are highly motile and spearhead colonial expansion. Explorers are characterized by high expression levels of flagellar genes, such as flagellin (hag), motA, fliI, flgK and sigD, hyperflagellation, decrease in ATP (adenosine-5'-triphosphate) levels, and increased resistance to antibiotics. Their tolerance to many antibiotics gives them the advantage of translocation through antibiotics-containing areas. This work gives new insights on the importance of cell differentiation and task distribution in colony morphogenesis and adaptation to antibiotics.


Passive transfer failure in horses: incidence and causative factors on a breeding farm.
A prospective study was performed to determine the incidence and associated maternal and managemental factors of failure of passive transfer (FPT) in foals on a breeding farm. The zinc sulfate turbidity test (ZSTT) and latex agglutination test (LAT) were compared for accuracy in estimating serum immunoglobulin (Ig)G of foals, as determined by single radial immunodiffusion (SRID). Complete past and present foaling histories of 136 Standardbred mares were obtained. All foalings were witnessed by farm attendants, and colostral samples were collected from mares within 2 hours after parturition. Foals that did not rise and nurse were supplemented with colostrum from the dam, using a bottle or nasogastric tube. Serum samples were prepared from foals and mares between 24 and 36 hours after parturition, and from some mares 45 to 90 days before parturition. Serum IgG concentrations of mares and foals and colostral whey were determined, using SRID. Serum IgG also was estimated in foals, using ZSTT and a commercially available LAT. Four of the 136 foals (2.9%) had FPT (serum IgG less than or equal to 400 mg/dl). Serum IgG concentrations in foals significantly correlated with colostral IgG (P less than 0.001). A significantly larger proportion of foals with FPT were bottle-fed their colostrum (P less than 0.01). Month of parturition, mare age, parity, number of barren seasons, incidence of assisted births or retained placenta, or prepartum serum IgG concentrations did not significantly affect colostral IgG concentrations or serum IgG concentrations in foals. As serum IgG concentrations in foals decreased and as colostral IgG concentrations decreased, the proportion of mares that prelactated significantly (P less than 0.01) increased.(ABSTRACT TRUNCATED AT 250 WORDS)


[Peculiarities of silicosis course in women engaged into metallurgy].
The study covers comparative analysis of silicosis course in males and females engaged into iron industry. Findings are more frequent combination of silicosis and chronic bronchitis in the females, without smoking habit. Some other differences in silicosis course and development were seen dependent on sex.


Perfluorinated compounds in seafood from coastal areas in China.
Diet is an important source of perfluorinated compound (PFC) exposure and seafood is an important diet component for coastal populations. Therefore, it is necessary to monitor the concentrations of PFCs in seafood. In this study, we measured thirteen PFCs in 47 fatty fish and 45 shellfish samples collected from six coastal provinces in China (Liaoning, Shandong, Jiangsu, Zhejiang, Fujian, and Guangdong), using an ultra-performance liquid chromatography and tandem mass spectrometry (UPLC-MS/MS). Perfluorooctanesulfonate (PFOS) was the dominant PFC in fatty fish which accounted for 38% of total PFCs, whereas perfluoroctanoic acid (PFOA) was the predominant PFC in shellfish. Concentrations of PFOS were ranged from less than 1.4 to 1627 pg/g wet weight in fatty fish, with the highest concentration in red drum from Jiangsu. Concentrations of PFOA in shellfish ranged from less than 5.4 to 7543 pg/g wet weight, with the maximum concentration found in briny clam also from Jiangsu. Compared with other studies, the PFC levels in seafood collected from our studied areas are relatively low. The estimated dietary intakes (EDI, pg/kg body weight /day) of PFOA, PFOS and total PFCs for the reference man (63 kg body weight) were calculated by multiplying the mean concentrations (pg/g wet weight) of PFOA, PFOS and total PFCs in six coastal provinces with the daily consumption data (g/day) from the fourth Chinese total diet study carried out in 2007. The highest EDI of PFOS and PFOA were found to be 694 pg/kg body weight/day and 914 pg/kg body weight/day in Guangdong and Jiangsu, respectively. However, the highest EDI of total PFCs was found in Fujian at 2513 pg/kg body weight/day. The EDI from seafood is much lower than the tolerable daily intake (TDI) recommended by the European Food Safety Authority in 2008 indicating low health risk of PFC exposure via eating seafood among the coastal populations in China.


The viscous consequence of different trends in clustering of 1,2-diol and 1,n-diol molecules.
This paper presents the molecular basis for the quite different behavior of the viscosity of 1,2- and 1,n-diols in dependence of the length of the alkyl part of the molecules of these compounds. The experimental data on the dipolar orientational effects revealed a decidedly different role of that part of the molecules in creating a microstructure of both the hydrogen-bonded liquids. In the case of 1,n-diols, an increase in the alkyl radical length, i.e. an increasing of the distance between the OH groups within the molecule, highly stimulates molecular self-assembly in form of gradually longer and wider ribbon-like clusters. This effect yields a quite important increase in the viscosity of 1,n-diols as n increases. In the case of 1,2-diols, due to gradual separation of the hydrophilic and hydrophobic parts of the molecules, the situation is quite different. Two OH groups situated on one of the ends of the hydrocarbon radical form the clusters of a micelle-like shape, however, the dipole moment is not compensated. Along with an increase in the hydrocarbon part in 1,2-diol molecules, one only observes an increase in the intermolecular consolidation within the micelle-like entities. This manifests as a gradual decrease in the polarity of these clusters. So, actually, there are no relevant reasons for essential differences of viscosities in the series of 1,2-diols.


Association between adiposity in midlife and older age and risk of diabetes in older adults.
Adiposity is a well-recognized risk factor for type 2 diabetes among young and middle-aged adults, but the relationship between body composition and type 2 diabetes is not well described among older adults. To examine the relationship between adiposity, changes in adiposity, and risk of incident type 2 diabetes in adults 65 years of age and older. Prospective cohort study (1989-2007) of 4193 men and women 65 years of age and older in the Cardiovascular Health Study. Measures of adiposity were derived from anthropometry and bioelectrical impedance data at baseline and anthropometry repeated 3 years later. Incident diabetes was ascertained based on use of antidiabetic medication or a fasting glucose level of 126 mg/dL or greater. Over median follow-up of 12.4 years (range, 0.9-17.8 years), 339 cases of incident diabetes were ascertained (7.1/1000 person-years). The adjusted hazard ratio (HR) (95% confidence interval [CI]) of type 2 diabetes for participants in the highest quintile of baseline measures compared with those in the lowest was 4.3 (95% CI, 2.9-6.5) for body mass index (BMI [calculated as weight in kilograms divided by height in meters squared]), 3.0 (95% CI, 2.0-4.3) for BMI at 50 years of age, 4.2 (95% CI, 2.8-6.4) for weight, 4.0 (95% CI, 2.6-6.0) for fat mass, 4.2 (95% CI, 2.8-6.2) for waist circumference, 2.4 (95% CI, 1.6-3.5) for waist-hip ratio, and 3.8 (95% CI, 2.6-5.5) for waist-height ratio. However, when stratified by age, participants 75 years of age and older had HRs approximately half as large as those 65 to 74 years of age. Compared with weight-stable participants (+/-2 kg), those who gained the most weight from 50 years of age to baseline (> or = 9 kg), and from baseline to the third follow-up visit (> or = 6 kg), had HRs for type 2 diabetes of 2.8 (95% CI, 1.9-4.3) and 2.0 (95% CI, 1.1-3.7), respectively. Participants with a greater than 10-cm increase in waist size from baseline to the third follow-up visit had an HR of type 2 diabetes of 1.7 (95% CI, 1.1-2.8) compared with those who gained or lost 2 cm or less. Among older adults, overall and central adiposity, and weight gain during middle age and after the age of 65 years are associated with risk of diabetes.


The patB gene of Dictyostelium discoideum encodes a P-type H(+)-ATPase isoform essential for growth and development under acidic conditions.
During growth and early development of Dictyostelium discoideum, the amoebae exhibit transient pH changes in their cytosol (pHi) and external medium which correlate with the extrusion of H+ from the cell by a plasma membrane pump. Moreover, the changes in pHi have been postulated to influence early prestalk/prespore differentiation during development. To learn more about the role of H+ fluxes in Dictyostelium, we cloned and analysed cDNAs of the gene patB, which appears to encode a P-type H(+)-ATPase. The patB ORF encodes a protein (termed PAT2) of 1058 amino acids with a calculated molecular mass of 117,460 Da. When aligned with other P-type ion-transport ATPases, PAT2 showed the greatest amino acid sequence identity with plasma membrane H(+)-ATPases of plants and fungi and considerably lower identity with other monovalent cation pumps and with Ca2+ pumps. Northern and Western analyses revealed that patB is expressed at very low levels in cells growing at neutral pH, but it is up-regulated rapidly and dramatically when the cells are shifted to an acidic medium. Immunofluorescence analysis indicated that PAT2 resides on the plasma membrane. When patB was disrupted by homologous recombination, the cells grew and developed normally at neutral and slightly alkaline pHs but they were unable to grow or develop at pH 5.0, and they slowly died. In growth medium at pH 6.8, patB+ and patB cells exhibited similar levels of vanadate-sensitive ATPase activity. However, when the cells were shifted to pH 5.0, this activity rapidly increased about twofold in the control cells but not in the mutant cells. Despite the lower ATPase activity in patB cells, they showed relatively normal H+ fluxes and only a slight decrease in pHi when incubated in acidic medium. Together, these results suggest that patB encodes an acid-inducible P-type H(+)-ATPase which is indispensable for the survival of Dictyostelium cells in moderately acidic external environments.


[Effect of moxibustion on the antibody-dependent cell-mediated cytotoxicity activity in peripheral blood of patients with Hashimoto's thyroiditis].
To investigate the immune regulating effect of moxibustion on the patients with Hashimoto's thyroiditis, 35 patients were treated by moxibustion and the changes of antibody-dependent cell-mediated cytotoxicity (ADCC) activity in the peripheral blood of the patients and their relationship with thyroid microsomal antibody (MCA) before and after treatment were observed. It was found that the activity of ADCC and the combining rate of MCA were higher than normal and they were closely related to each other. After treatment, both of them declined remarkably and the relationship between them remained the same. The result indicated that the treatment of Hashimoto's thyroiditis by moxibustion might well be related to its effect of decreasing the activity of ADCC and the titer of anti-thyroid antibodies.


Hyperactive Cdc2 kinase interferes with the response to broken replication forks by trapping S.pombe Crb2 in its mitotic T215 phosphorylated state.
Although it is well established that Cdc2 kinase phosphorylates the DNA damage checkpoint protein Crb2(53BP1) in mitosis, the full impact of this modification is still unclear. The Tudor-BRCT domain protein Crb2 binds to modified histones at DNA lesions to mediate the activation of Chk1 by Rad3ATR kinase. We demonstrate here that fission yeast cells harbouring a hyperactive Cdc2CDK1 mutation (cdc2.1w) are specifically sensitive to the topoisomerase 1 inhibitor camptothecin (CPT) which breaks DNA replication forks. Unlike wild-type cells, which delay only briefly in CPT medium by activating Chk1 kinase, cdc2.1w cells bypass Chk1 to enter an extended cell-cycle arrest which depends on Cds1 kinase. Intriguingly, the ability to bypass Chk1 requires the mitotic Cdc2 phosphorylation site Crb2-T215. This implies that the presence of the mitotic phosphorylation at Crb2-T215 channels Rad3 activity towards Cds1 instead of Chk1 when forks break in S phase. We also provide evidence that hyperactive Cdc2.1w locks cells in a G1-like DNA repair mode which favours non-homologous end joining over interchromosomal recombination. Taken together, our data support a model such that elevated Cdc2 activity delays the transition of Crb2 from its G1 to its G2 mode by blocking Srs2 DNA helicase and Casein Kinase 1 (Hhp1).


Comparative analyses reveal potential genetic mechanisms for high-altitude adaptation of Schizopygopsis fishes based on chromosome-level genomes.
The schizothoracine fishes, widely distributed in the Qinghai-Tibetan Plateau and its adjacent areas, are considered as ideal models for investigation of high-altitude adaptation. Schizophygopsis are one group of the highly specialized schizothoracine fishes, and the genetic basis for their high-altitude adaptation is poorly understood. In this study, we performed comparative genomics analyses to investigate the potential genetic mechanisms for high-altitude adaptation of Schizopygopsis malacanthus and Schizopygopsis pylzovi based on the chromosome-level genomes. Functional enrichment analysis revealed that many expanded gene families in Schizopygopsis were associated with immune response while many contracted gene families were functionally associated with olfaction. Among the 123 positively selected genes (PSGs), angpt2a was detected in HIF-1 signaling pathway and possibly related to the hypoxia adaptation of Schizopygopsis. Furthermore, two PSGs cox15 and ndufb10 were distributed in thermogenesis, and there was a Schizopygopsis-specific missense mutation in cox15 (Gln115Glu), which possibly contributed to the cold temperature adaptation of the Schizopygopsis. Kyoto Encyclopedia of Genes and Genomes enrichment of the PSGs revealed three significant pathways including metabolic pathways, cell cycle, and homologous recombination and Gene Ontology enrichment analysis of the PSGs revealed several categories associated with DNA repair, cellular response to DNA damage stimulus, and metabolic process. Chromosome-scale characterization of olfactory receptor (OR) repertoires indicated that Schizopygopsis had the least number of OR genes, and the OR gene contraction was possibly caused by the limited food variety and the environmental factors such as lower air pressure, lower humidity, and lower temperature. Our study will help expand our understanding of the potential adaptive mechanism of Schizopygopsis to cope with the high-altitude conditions.


A Mass Spectrometric Assay of METTL3/METTL14 Methyltransferase Activity.
A variety of covalent modifications of RNA have been identified and demonstrated to affect RNA processing, stability, and translation. Methylation of adenosine at the N6 position (m6A) in messenger RNA (mRNA) is currently the most well-studied RNA modification and is catalyzed by the RNA methyltransferase complex METTL3/METTL14. Once generated, m6A can modulate mRNA splicing, export, localization, degradation, and translation. Although potent and selective inhibitors exist for several members of the Type I S-adenosylmethionine (SAM)-dependent methyltransferase family, no inhibitors have been reported for METTL3/METTL14 to date. To facilitate drug discovery efforts, a sensitive and robust mass spectrometry-based assay for METTL3/METTL14 using self-assembled monolayer desorption/ionization (SAMDI) technology has been developed. The assay uses an 11-nucleotide single-stranded RNA compared to a previously reported 27-nucleotide substrate. IC50 values of mechanism-based inhibitors S-adenosylhomocysteine (SAH) and sinefungin (SFG) are comparable between the SAMDI and radiometric assays that use the same substrate. This work demonstrates that SAMDI technology is amenable to RNA substrates and can be used for high-throughput screening and compound characterization for RNA-modifying enzymes.