The Role of the Artery of Ligamentum Teres in Revascularization in Legg-Calve-Perthes Disease.
A recent cadaveric study supported that most immature hips are supplied by the artery of ligamentum teres and suggested this medial vascular source may influence the pattern of revascularization in Legg-Calve-Perthes disease (LCPD). The purposes of this study were to characterize the perfusion pattern of the capital femoral epiphysis and determine the role of the artery of ligamentum teres in early revascularization of LCPD. Retrospective review of perfusion magnetic resonance imaging (pMRI) from 64 hips in early stage LCPD (Waldenström stage I to IIa) was performed. Two independent graders categorized perfusion pattern based on the presence of perfusion medially (from artery of ligamentum teres) and/or laterally (from the medial femoral circumflex artery) on coronal and sagittal MRI series: type 1-lateral perfusion only, type 2-separate medial and lateral perfusion, or type 3-coalescent medial and lateral perfusion. Lateral pillar classification was obtained for hips that reached mid-fragmentation. We identified 64 patients (75% male) with mean age at diagnosis of 8.5±2.1 years. 36% (23/64) hips underwent pMRI during stage I and 64% (41/64) during stage IIa. pMRI revealed separate and distinct medial and lateral sources of perfusion (type 2) in 50% (32/64) hips. In stage I, the distribution of type 1/2/3 hips was found to be 26%/52%/22%. However, in stage IIa there was a nonsignificant trend toward greater coalescence of the medial and lateral perfusion with a distribution of type 1/2/3 of 7%/49%/44% (P=0.07). There was a nonsignificant trend toward weak negative linear correlation between lower initial perfusion grade and worsened lateral pillar classification at mid-fragmentation (r=-0.25, P=0.05). The presence of separate and distinct areas of perfusion of medial and lateral capital femoral epiphysis provides further evidence of the role of the ligamentum teres vessels in revascularization during the early stages of LCPD. The changes in perfusion pattern with disease progression likely reflect that medial femoral circumflex artery and ligamentum teres vessel revascularization occur separately, but ultimately coalesce posteriorly over time. Level II-prognostic study.


Language identification from visual-only speech signals.
Our goal in the present study was to examine how observers identify English and Spanish from visual-only displays of speech. First, we replicated the recent findings of Soto-Faraco et al. (2007) with Spanish and English bilingual and monolingual observers using different languages and a different experimental paradigm (identification). We found that prior linguistic experience affected response bias but not sensitivity (Experiment 1). In two additional experiments, we investigated the visual cues that observers use to complete the language-identification task. The results of Experiment 2 indicate that some lexical information is available in the visual signal but that it is limited. Acoustic analyses confirmed that our Spanish and English stimuli differed acoustically with respect to linguistic rhythmic categories. In Experiment 3, we tested whether this rhythmic difference could be used by observers to identify the language when the visual stimuli is temporally reversed, thereby eliminating lexical information but retaining rhythmic differences. The participants performed above chance even in the backward condition, suggesting that the rhythmic differences between the two languages may aid language identification in visual-only speech signals. The results of Experiments 3A and 3B also confirm previous findings that increased stimulus length facilitates language identification. Taken together, the results of these three experiments replicate earlier findings and also show that prior linguistic experience, lexical information, rhythmic structure, and utterance length influence visual-only language identification.


[Patient with sudden cardiac arrest, peripartum cardiomyopathy and pulmonary embolism].
Peripartum cardiomyopathy is a type of dilatated cardiomyopathy, occuring with symptoms of heart failure (HF) during last month of pregnancy or within 5 months after labour. Authors are presenting the case of patient admitted to hospital primary with diagnosis of non-high risk pulmonary embolism 6 weeks after delivery, who developed episode of sudden cardiac death followed by symptoms of cardiogenic shock. Peripartum cardiomyopathy was additionally diagnosed. After HF treatment with bromocriptine supply, gradual clinical improvement was achieved. The patient was discharged after 15 days of hospitalisation with diagnosis of peripartum cardiomyopathy with non-high risk pulmonary embolism.


Immunopathogenesis, immunology and assessment of activity of sarcoidosis.
To this point we have attempted to summarize what we believe to be the most exciting new findings relative to sarcoidosis, along with providing data in support of those comments. We wish to conclude this brief review by sharing some anecdotal thoughts (no benefit of controlled clinical trials) concerning how the individual practitioner may apply this "research" information in caring for patients. In newly diagnosed patients with sarcoidosis, especially with widespread use of transbronchial lung biopsy, we would recommend BAL at that time because it can provide very valuable information with minimal increased morbidity or cost. BAL, in combination with SACE and possibly gallium scan (not required in all), along with the usual measures such as pulmonary function studies, chest roentgenograms (all of which are already commonly done) permit the clinician to make an accurate conclusion about disease activity in the lungs. Because there are no published therapeutic trials of corticosteroids in which the ultimate prognosis of pulmonary sarcoidosis has been improved, the decision to initiate therapy in these patients remains a clinical one and is commonly based upon practitioner experience rather than a particular laboratory test per se. Once that decision has been made, a decision to repeat any "research" studies would have to be individualized. Clearly serial SACE measures are relatively inexpensive and without morbidity. Moreover, despite their lack of "diagnostic value" serial measurements in known sarcoidosis have been shown to be very valuable in assessing disease activity and may therefore provide the clinician with a valuable aid to monitoring steroid treatment. A less common and much more perplexing circumstance occurs when evaluating patients with sarcoidosis established for several years present with evidence of progressive or chronic pulmonary involvement and dyspnea. In such individuals a measure of activity would be very helpful because the clinician would like to avoid steroids in persons with end stage and irreversible fibrosis and consider their use in those individuals in whom there is evidence of continued disease activity. In this instance, gallium scan, SACE and even BAL may all prove valuable. When we have found SACE elevated and gallium scan positive, we have assumed the disease to be active (even if the chest X-rays show stability).(ABSTRACT TRUNCATED AT 400 WORDS)


Needle aspiration cytologic examination in the management of suspicious lesions of the breast.
The experience of one surgeon with needle aspiration cytologic examination used as an office procedure in the management of suspicious solid lesions of the breast over a period of seven years in presented. Three hundred and thirty-five aspirations were performed with the diagnosis of carcinoma subsequently established in 126 or 37 per cent of the patients. In 79 of these patients (63 per cent) carcinoma was diagnosed preoperatively by positive needle aspiration cytologic examination only. There were no false-positive results. Thus, this group of patients was spared time and added expense of open biopsy before definitive surgical treatment was instituted.


Quantum collapse and the second law of thermodynamics.
A heat engine undergoes a cyclic operation while in equilibrium with the net result of conversion of heat into work. Quantum effects such as superposition of states can improve an engine's efficiency by breaking detailed balance, but this improvement comes at a cost due to excess entropy generated from collapse of superpositions on measurement. We quantify these competing facets for a quantum ratchet composed of an ensemble of pairs of interacting two-level atoms. We suggest that the measurement postulate of quantum mechanics is intricately connected to the second law of thermodynamics. More precisely, if quantum collapse is not inherently random, then the second law of thermodynamics can be violated. Our results challenge the conventional approach of simply quantifying quantum correlations as a thermodynamic work deficit.


Intraspecific Diversity of Microbial Anti-Inflammatory Molecule (MAM) from Faecalibacterium prausnitzii.
The commensal bacterium Faecalibacterium prausnitzii has unique anti-inflammatory properties, at least some of which have been attributed to its production of MAM, the Microbial Anti-inflammatory Molecule. Previous phylogenetic studies of F. prausnitzii strains have revealed the existence of various phylogroups. In this work, we address the question of whether MAMs from different phylogroups display distinct anti-inflammatory properties. We first performed wide-scale identification, classification, and phylogenetic analysis of MAM-like proteins encoded in different genomes of F. prausnitzii. When combined with a gene context analysis, this approach distinguished at least 10 distinct clusters of MAMs, providing evidence for functional diversity within this protein. We then selected 11 MAMs from various clusters and evaluated their anti-inflammatory capacities in vitro. A wide range of anti-inflammatory activity was detected. MAM from the M21/2 strain had the highest inhibitory effect (96% inhibition), while MAM from reference strain A2-165 demonstrated only 56% inhibition, and MAM from strain CNCM4541 was almost inactive. These results were confirmed in vivo in murine models of acute and chronic colitis. This study provides insights into the family of MAM proteins and generates clues regarding the choice of F. prausnitzii strains as probiotics for use in targeting chronic inflammatory diseases.


Targeting core (mutated) pathways of high-grade gliomas: challenges of intrinsic resistance and drug efflux.
High-grade gliomas are the most common type of primary brain tumor and are among the most lethal types of human cancer. Most patients with a high-grade glioma have glioblastoma multiforme (GBM), the most malignant glioma subtype that is associated with a very aggressive disease course and short overall survival. Standard treatment of newly diagnosed GBM involves surgery followed by chemoradiation with temozolomide. However, despite this extensive treatment the mean overall survival is still only 14.6 months and more effective treatments are urgently needed. Although different types of GBMs are indistinguishable by histopathology, novel molecular pathological techniques allow discrimination between the four main GBM subtypes. Targeting the aberrations in the molecular pathways underlying these subtypes is a promising strategy to improve therapy. In this article, we will discuss the potential avenues and pitfalls of molecularly targeted therapies for the treatment of GBM.


Heat shock protein-GP96 as an innate sensor of damage and activator of autoreactive NKT and regulatory T cells during liver regeneration.
Tissue disintegration after injury leads, in the endoplasmic reticulum (ER), to activation of adaptive pathways known as the ER stress response. It is directed to the correction of unfolded proteins and to the activation of proteasome-dependent ER-associated degradation of the misfolded proteins, but induces also a rapid activation of natural and adaptive immunity, since a ER resident heat shock protein-gp96 acts not only as a molecular chaperone, but also as a strong adjuvant, able to cross-present the antigenic peptides onto MHC class I or MHC class II pathways. Analyzing its potential role in processes of normal growth, in mice subjected to 1/3 partial hepatectomy (pHx) we determined the tissue expression of gp96 protein and mRNA in regenerating liver, thymus and spleen, determining simultaneously the phenotypic profile and spontaneous cytotoxic activity of intrahepatic and splenic mononuclear lymphatic cells (MNLC) against NKT- and NK-cells sensitive targets (syngeneic thymocytes and YAC-1) in wild, perforin and FasL deficient mice. The data have shown that pHx induces fast overexpression of gp96 protein and mRNA in hepatocytes, spleen and thymus, with accumulation of CD3intermediate/NK1.1+/CD69+ cells (liver) and Foxp3+CD4+CD25+ cells (liver and thymus). Simultaneously, intrahepatic MNLC acquired the FasL-dependent cytotoxic potential against NKT-sensitive targets and both, intrahepatic and splenic MNLC, acquired the perforin-dependent cytotoxic potential against NK-sensitive targets, implying that during the disturbance of morphostasis gp96 serves as a natural adjuvant for chaperoning antigenic self peptides into the immune surveillance pathways, resulting in activation of autoreactive NKT and regulatory cells, as well as NK cells. Moreover, cell cycle analysis revealed that G2+M phase of regenerating hepatocytes in PKO mice was translocated from the 1st to the 7th p. o. day, as well as that hepatocytes from FasL deficient mice were arrested in G0/G1 phase.


Biochemical abnormalities in Pearson syndrome.
Pearson marrow-pancreas syndrome is a multisystem mitochondrial disorder characterized by bone marrow failure and pancreatic insufficiency. Children who survive the severe bone marrow dysfunction in childhood develop Kearns-Sayre syndrome later in life. Here we report on four new cases with this condition and define their biochemical abnormalities. Three out of four patients presented with failure to thrive, with most of them having normal development and head size. All patients had evidence of bone marrow involvement that spontaneously improved in three out of four patients. Unique findings in our patients were acute pancreatitis (one out of four), renal Fanconi syndrome (present in all patients, but symptomatic only in one), and an unusual organic aciduria with 3-hydroxyisobutyric aciduria in one patient. Biochemical analysis indicated low levels of plasma citrulline and arginine, despite low-normal ammonia levels. Regression analysis indicated a significant correlation between each intermediate of the urea cycle and the next, except between ornithine and citrulline. This suggested that the reaction catalyzed by ornithine transcarbamylase (that converts ornithine to citrulline) might not be very efficient in patients with Pearson syndrome. In view of low-normal ammonia levels, we hypothesize that ammonia and carbamylphosphate could be diverted from the urea cycle to the synthesis of nucleotides in patients with Pearson syndrome and possibly other mitochondrial disorders.


Geldanamycin activates Hsp70 response and attenuates okadaic acid-induced cytotoxicity in human retinal pigment epithelial cells.
Reversible protein phosphorylation regulates the biological activities of many human proteins involved in crucial cellular processes, e.g., protein-protein interactions, cell signaling, gene transcription, cell growth, and death. A malfunction of cellular homeostasis in retinal pigment epithelial (RPE) cells is involved in the age-related retinal degeneration. In this study, we examined cytotoxicity in human RPE cells subjected to the protein phosphatase inhibitor, okadaic acid (OA). Moreover, the influence of Hsp90 inhibitor geldanamycin (GA), a benzoquinone ansamycin, in cytoprotection was assessed. Hsp70 protein levels were analyzed by Western blot. Cellular viability was determined by LDH and MTT assays. To study apoptotic cell death, caspase-3 enzyme activity was measured by assaying the cleavage of a fluorescent peptide substrate and Hoechst dye was used to visualize nuclear morphology. OA treatment caused morphological changes and induced cytotoxicity by caspase-3-independent manner in the RPE cells. No evidence of nuclear fragmentation was observed in response to OA. Interestingly, GA treatment accumulated Hsp70 protein and attenuated OA-induced cytotoxicity. This study suggests that Hsp70 and Hsp90 are closely related to cytoprotection of RPE cells in response to protein phosphatase inhibition.


Self-reported health: a study of older adults from a developing country--Nepal.
Self-reported health is an easy measure of overall health and is useful in identifying persons at risk of a decline in health and the risk of disability in older adults. The purpose of this paper is to identify the correlates of self-reported health for older adults in Nepal. The subjects (N = 137) were members of the Chhetri caste who were 60 years or older (mean age 69.1 +/- 7.2 years; 54% women) and living in Katmandu City. Data were collected by face-to face interviews using three self-reported health questions. A single scale was developed from the three self-reported questions. Bivariate analysis and multiple regression analysis were used to analyze the results. The findings show a correlation between self-reported health and sex, household status, living arrangements, social participation, chronic health problems, functional disability in activities of daily living (ADL), employment status, and economic satisfaction. Results of multiple regression analysis indicate that factors for self-reported health among older adults are chronic health problems, economic satisfaction, functional disability in ADLs, and social participation.


Brown adipose tissue: The heat is on the heart.
The study of brown adipose tissue (BAT) has gained significant scientific interest since the discovery of functional BAT in adult humans. The thermogenic properties of BAT are well recognized; however, data generated in the last decade in both rodents and humans reveal therapeutic potential for BAT against metabolic disorders and obesity. Here we review the current literature in light of a potential role for BAT in beneficially mediating cardiovascular health. We focus mainly on BAT's actions in obesity, vascular tone, and glucose and lipid metabolism. Furthermore, we discuss the recently discovered endocrine factors that have a potential beneficial role in cardiovascular health. These BAT-secreted factors may have a favorable effect against cardiovascular risk either through their metabolic role or by directly affecting the heart.


JNK signaling is the shared pathway linking neuroinflammation, blood-brain barrier disruption, and oligodendroglial apoptosis in the white matter injury of the immature brain.
White matter injury is the major form of brain damage in very preterm infants. Selective white matter injury in the immature brain can be induced by lipopolysaccharide (LPS)-sensitized hypoxic-ischemia (HI) in the postpartum (P) day 2 rat pups whose brain maturation status is equivalent to that in preterm infants less than 30 weeks of gestation. Neuroinflammation, blood-brain barrier (BBB) damage and oligodendrocyte progenitor apoptosis may affect the susceptibility of LPS-sensitized HI in white matter injury. c-Jun N-terminal kinases (JNK) are important stress-responsive kinases in various forms of insults. We hypothesized that LPS-sensitized HI causes white matter injury through JNK activation-mediated neuroinflammation, BBB leakage and oligodendroglial apoptosis in the white matter of P2 rat pups. P2 pups received LPS (0.05 mg/kg) or normal saline injection followed by 90-min HI. Immunohistochemistry and immunoblotting were used to determine microglia activation, TNF-α, BBB damage, cleaved caspase-3, JNK and phospho-JNK (p-JNK), myelin basic protein (MBP), and glial fibrillary acidic protein (GFAP) expression. Immunofluorescence was performed to determine the cellular distribution of p-JNK. Pharmacological and genetic approaches were used to inhibit JNK activity. P2 pups had selective white matter injury associated with upregulation of activated microglia, TNF-α, IgG extravasation and oligodendroglial progenitor apoptosis after LPS-sensitized HI. Immunohistochemical analyses showed early and sustained JNK activation in the white matter at 6 and 24 h post-insult. Immunofluorescence demonstrated upregulation of p-JNK in activated microglia, vascular endothelial cells and oligodendrocyte progenitors, and also showed perivascular aggregation of p-JNK-positive cells around the vessels 24 h post-insult. JNK inhibition by AS601245 or by antisense oligodeoxynucleotides (ODN) significantly reduced microglial activation, TNF-α immunoreactivity, IgG extravasation, and cleaved caspase-3 in the endothelial cells and oligodendrocyte progenitors, and also attenuated perivascular aggregation of p-JNK-positive cells 24 h post-insult. The AS601245 or JNK antisense ODN group had significantly increased MBP and decreased GFAP expression in the white matter on P11 than the vehicle or scrambled ODN group. LPS-sensitized HI causes white matter injury through JNK activation-mediated upregulation of neuroinflammation, BBB leakage and oligodendrocyte progenitor apoptosis in the immature brain.


[Research Progress of Iron Metabolism in Disease Progression and Drug Resistance of Multiple Myeloma--Review].
Iron metabolism is involved in the development and drug resistance of many malignancies, including multiple myeloma (MM). Based on recent studies on iron metabolism and MM, this paper reviews the relationship between iron metabolism and disease process of MM in terms of iron overload leading to ferroptosis in MM cells, the role of iron deficiency in oxidative respiration and proliferation of MM cells, and the interaction between ferroptosis and autophagy in the disease process. The mechanisms by which iron metabolism-related substances lead to MM cells' resistance to proteasome inhibitors (PI) through inducing redox imbalance and M2 macrophage polarization are also briefly described, aiming to provide a theoretical basis for the application of iron metabolism-related drugs to the clinical treatment of MM patients.


Novel method for analyzing locomotor ability after spinal cord injury in rats: technical note.
In the research for the treatment of spinal cord injury (SCI), the evaluation of motor function in model rats must be as objective, noninvasive, and ethical as possible. The maximum speed and acceleration of a mouse measured using a SCANET system were previously reported to vary significantly according to severity of SCI. In the present study, the motor performance of SCI model rats was examined with SCANET and assessed for Basso-Beattie-Bresnahan (BBB) score to determine the usefulness of the SCANET system in evaluating functional recovery after SCI. Maximum speed and acceleration within the measurement period correlated significantly with BBB scores. Furthermore, among several phased kinematic factors used in BBB scores, the capability of "plantar stepping" was associated with a drastic increase in maximum speed and acceleration after SCI. Therefore, evaluation of maximum speed and acceleration using a SCANET system is a useful method for rat models of SCI and can complement open field scoring scales.


Snack Food, Satiety, and Weight.
In today's society, snacking contributes close to one-third of daily energy intake, with many snacks consisting of energy-dense and nutrient-poor foods. Choices made with regard to snacking are affected by a multitude of factors on individual, social, and environmental levels. Social norms, for example, that emphasize healthful eating are likely to increase the intake of nutrient-rich snacks. In addition, satiety, the feeling of fullness that persists after eating, is an important factor in suppressing overconsumption, which can lead to overweight and obesity. Thus, eating snacks between meals has the potential to promote satiety and suppress overconsumption at the subsequent meal. Numerous studies have explored the relation between snack foods and satiety. These studies concluded that whole foods high in protein, fiber, and whole grains (e.g., nuts, yogurt, prunes, and popcorn) enhance satiety when consumed as snacks. Other foods that are processed to include protein, fiber, or complex carbohydrates might also facilitate satiety when consumed as snacks. However, studies that examined the effects of snack foods on obesity did not always account for satiety and the dietary quality and portion size of the snacks consumed. Thus, the evidence concerning the effects of snack foods on obesity has been mixed, with a number of interventional and observational studies not finding a link between snack foods and increased weight status. Although further prospective studies are warranted to conclusively determine the effects of snack foods on obesity risk, the consumption of healthful snacks likely affects satiety and promotes appetite control, which could reduce obesity.


Effects of intraluminal distention and decompression on microvascular permeability and hemodynamics of the equine jejunum.
To determine whether intraluminal distention and subsequent decompression of the equine jejunum affects intestinal blood flow, hemodynamics, and microvascular permeability. 5 healthy adu t horses. Horses were anesthestized and underwent exploratory laparotomy. Two jejunal segments were identified as sham-operated or instrumented segments. After baseline values were obtained, intraluminal distention was created in the experimental segment to induce an ntraluminal pressure of 18 cm H2O. After 120 minutes of distention, the intestine was decompressed for 120 minutes. Mesenteric blood flow, oxygen delivery, oxygen consumption, microvascular permeability, wet weight-to-dry weight ratio, neutrophil infiltration, and vascular resistance were determined and comparisons made among control, sham-operated, and experimental segments. Mean jejunal blood flow was 21.4 ml/min per kg. There was a significant decrease in mesenteric bood flow to the distended intestine (13.4 ml/min per kg). Blood flow increased significantly during the decompression period (340% of baseline blood flow). Intraluminal distention and subsequent decompression resulted in a significant increase in microvascular permeability, as determined by the osmotic reflection coefficient. Oxygen delivery and oxygen content decreased significantly during the distention period and increased during decompression. Morphologic evaluation revealed a significant increase in edema and neutrophil infiltration after distention and decompression, compared with results for the sham-operated or control segments. Intraluminal distention and decompression of the equine jejunum results in low-flow ischemia and edema, which may contribute to adhesions and ileus in the postoperative period after surgery for obstructions of the small intestines.


[If I were to wake up clumsy and shaking (10 years later)].
The diagnosis and treatment of early Parkinson's disease (PD) has undergone considerable changes in the last 10 years. Identification of genes responsible for familial forms of the disease as well as the possibility of detecting dopaminergic dysfunction before the appearance of motor symptoms has opened up the possibility of presymptomatic diagnosis. The concept of PD is changing. Therapeutic strategies are quite different. Several studies support the recommendation of not delaying the onset of dopaminergic therapies. Nonetheless, neuroprotection still has not been totally demonstrated and this continues to be an objective of priority.


5-Fluorouracil induced Fas upregulation associated with apoptosis in liver metastases of colorectal cancer patients.
In vitro, thymidylate synthase (TS) inhibition by 5-fluorouracil (5-FU) induces thymineless apoptosis possibly via Fas receptor Fas ligand interactions and cell-cycle arrest. In colorectal cancer patients we evaluated whether 5-FU administration also resulted in apoptosis and cell-cycle arrest and which proteins might be involved. Biopsy specimens were taken from 36 patients 2, 22 or 46 hours after administration of 500 mg/m2 5-FU, and from 12 control patients who did not receive 5-FU. In frozen tissue-sections from liver metastases immunohistochemistry was performed with antibodies directed against p53, p21, E2F2, Rb, Ki67 and TS (cell-cycle related) and bax, BCL-2, BCL-x, mcl-1, PARP, caspase-3, Fas receptor and Fas ligand (apoptosis related). Apoptosis was determined by M30 immunostaining, which recognises a cleavage product of cytokeratin 18. Fas receptor expression was 50% higher (P = 0.036) 46 hours after 5-FU administration compared to the control group. This was associated with a 12% increase (P < 0.02) in M30 positive tumour cells and with elevation of caspase-3 and PARP expression. The expression of Ki67 and E2F2 was 30% lower after 46 hours compared to the control group, whereas TS was 56% lower after 2 hours and 32% higher again after 46 hours. No differences in the expression of the other proteins were found. These results suggest that 5-FU decreases proliferation status and induces apoptosis possibly via the Fas pathway. Since Fas mediated cell killing is important for cytotoxic T cells this indicates that clinical studies combining immunotherapy for activation of T cells and chemotherapy using 5-FU might be very effective.


Plant organellar DNA primase-helicase synthesizes RNA primers for organellar DNA polymerases using a unique recognition sequence.
DNA primases recognize single-stranded DNA (ssDNA) sequences to synthesize RNA primers during lagging-strand replication. Arabidopsis thaliana encodes an ortholog of the DNA primase-helicase from bacteriophage T7, dubbed AtTwinkle, that localizes in chloroplasts and mitochondria. Herein, we report that AtTwinkle synthesizes RNA primers from a 5'-(G/C)GGA-3' template sequence. Within this sequence, the underlined nucleotides are cryptic, meaning that they are essential for template recognition but are not instructional during RNA synthesis. Thus, in contrast to all primases characterized to date, the sequence recognized by AtTwinkle requires two nucleotides (5'-GA-3') as a cryptic element. The divergent zinc finger binding domain (ZBD) of the primase module of AtTwinkle may be responsible for template sequence recognition. During oligoribonucleotide synthesis, AtTwinkle shows a strong preference for rCTP as its initial ribonucleotide and a moderate preference for rGMP or rCMP incorporation during elongation. RNA products synthetized by AtTwinkle are efficiently used as primers for plant organellar DNA polymerases. In sum, our data strongly suggest that AtTwinkle primes organellar DNA polymerases during lagging strand synthesis in plant mitochondria and chloroplast following a primase-mediated mechanism. This mechanism contrasts to lagging-strand DNA replication in metazoan mitochondria, in which transcripts synthesized by mitochondrial RNA polymerase prime mitochondrial DNA polymerase γ.


Proteomics analysis of chinese hamster ovary cells undergoing apoptosis during prolonged cultivation.
The degradation of environmental conditions, such as nutrient depletion and accumulation of toxic waste products over time, often lead to premature apoptotic cell death in mammalian cell cultures and suboptimal protein yield. Although apoptosis has been extensively researched, the changes in the whole cell proteome during prolonged cultivation, where apoptosis is a major mode of cell death, have not been examined. To our knowledge, the work presented here is the first whole cell proteome analysis of non-induced apoptosis in mammalian cells. Flow cytometry analyses of various activated caspases demonstrated the onset of apoptosis in Chinese hamster ovary cells during prolonged cultivation was primarily through the intrinsic pathway. Differential in gel electrophoresis proteomic study comparing protein samples collected during cultivation resulted in the identification of 40 differentially expressed proteins, including four cytoskeletal proteins, ten chaperone and folding proteins, seven metabolic enzymes and seven other proteins of varied functions. The induction of seven ER chaperones and foldases is a solid indication of the onset of the unfolded protein response, which is triggered by cellular and ER stresses, many of which occur during prolonged batch cultures. In addition, the upregulation of six glycolytic enzymes and another metabolic protein emphasizes that a change in the energy metabolism likely occurred as culture conditions degraded and apoptosis advanced. By identifying the intracellular changes during cultivation, this study provides a foundation for optimizing cell line-specific cultivation processes, prolonging longevity and maximizing protein production.


A novel disorder reveals clathrin heavy chain-22 is essential for human pain and touch development.
Congenital inability to feel pain is very rare but the identification of causative genes has yielded significant insights into pain pathways and also novel targets for pain treatment. We report a novel recessive disorder characterized by congenital insensitivity to pain, inability to feel touch, and cognitive delay. Affected individuals harboured a homozygous missense mutation in CLTCL1 encoding the CHC22 clathrin heavy chain, p.E330K, which we demonstrate to have a functional effect on the protein. We found that CLTCL1 is significantly upregulated in the developing human brain, displaying an expression pattern suggestive of an early neurodevelopmental role. Guided by the disease phenotype, we investigated the role of CHC22 in two human neural crest differentiation systems; human induced pluripotent stem cell-derived nociceptors and TRKB-dependant SH-SY5Y cells. In both there was a significant downregulation of CHC22 upon the onset of neural differentiation. Furthermore, knockdown of CHC22 induced neurite outgrowth in neural precursor cells, which was rescued by stable overexpression of small interfering RNA-resistant CHC22, but not by mutant CHC22. Similarly, overexpression of wild-type, but not mutant, CHC22 blocked neurite outgrowth in cells treated with retinoic acid. These results reveal an essential and non-redundant role for CHC22 in neural crest development and in the genesis of pain and touch sensing neurons.


[Two types of diaphragmatic plication; therapeutic plication and prophylactic plication].
Plication of the hemidiaphragm is effective to relieve respiratory symptoms such as exertional dyspnea and orthopnea of the patients with unilateral diaphragmatic paralysis. We have experiences of 2 types of diaphragmatic plication;one for the symptomatic patients with unilateral diaphragmatic paralysis, and the other for the patients who underwent phrenicotomy during extended surgery for a recurrent thymoma. The former plication was performed for a therapeutic purpose to alleviate the patient's symptoms. The latter was performed for a prophylactic purpose to keep the patient free from possible symptoms or postoperative morbidity after phrenicotomy. Case 1:A 65-year-old female with an eventration of the right diaphragm underwent a sternal resection and reconstruction for a large desmoid tumor. After continuous mechanical ventilation for 3 months, she was still under noninvasive positive pressure ventilation( NPPV) for 24 hours a day. Plication of the right hemidiaphragm was performed and the patient was successfully weaned from NPPV. Case 2:A 41-year-old female with myasthenia gravis underwent a surgery for a recurrent thymoma. Left phrenicotomy was necessary to achieve a complete resection of the tumor. Plication of the left diaphragm was performed during the tumor resection to prevent possible respiratory complications. Postoperative course was uneventful, and her vital capacity was well maintained.


Gonadotropin-releasing hormone-regulated prohibitin mediates apoptosis of the gonadotrope cells.
GnRH regulates circulating levels of the gonadotropins but has also been implicated in establishing the gonadotrope cell population. Consistent with this, GnRH induces proliferation of partially differentiated gonadotropes, while reducing the numbers of fully differentiated cells. We have previously reported that the proapoptotic protein, prohibitin (PHB) is expressed more abundantly in gonadotrope-derived LβT2 cells than in partially differentiated αT3-1 gonadotrope precursor cells, suggesting a possible role for PHB in GnRH-induced apoptosis. We show here that PHB is required for GnRH-induced apoptosis in mature gonadotropes. PHB expression and activity are regulated by GnRH: its transcription is via c-Jun NH2-terminal kinase, whereas its nuclear export follows activation of ERK. Moreover, PHB levels are down-regulated by microRNA27, which is expressed at lower levels in mature gonadotropes, possibly explaining the switch to an apoptotic response with development. PHB is required for mitochondrial import of the proapoptotic BAX, whose expression is also induced by GnRH-activated c-Jun NH2-terminal kinase, as is expression of the BH3-only protein, HRK, and this too plays a role in GnRH-induced apoptosis. Finally, we show that gonadotrope-specific PHB-knockout mice display reproductive abnormalities, including a larger gonadotrope population, increased LH levels, reduced fertility, and altered gonad development. We thus demonstrate a role for PHB in GnRH-induced cell death in mature gonadotropes, which is crucial for the normal development and function of the reproductive axis.


Lumbar puncture in neonates under and over 72 hours of age.
To identify the criteria for performing a spinal tap in neonates with suspected sepsis and compare the findings in infants under and over the age of 72 hours. Analytical. Neonatal Ward of Taleghani Medical Centre, Shaheed Beheshti University of Medical Sciences, Evin,Tehran. Study period was of one year, from March 2003 to February 2004. All newborns subjected to a lumbar puncture during the first 24 hours of admission were enrolled in the study. Results of cerebrospinal fluid analysis were scrutinized. There were 380 neonates in the study. Majority i.e. 248 infants were aged<72 hours. Meningitis was diagnosed in 22 cases, (5.8%). There was no significant difference in the incidence of meningitis between neonates under and over 72 hours of age, (6.9% vs. 3.8%, p-value>0.5). Comparison of risk factors, clinical manifestations, and routine laboratory investigations revealed no significant difference in babies with or without meningitis. The findings of this series do not support a selective approach for performing a lumbar puncture in a possibly septic newborn. A lumbar puncture is recommended in all neonates with suspected sepsis regardless of age, risk factors, or specific neurological manifestations.


Mean shift trackers with cross-bin metrics.
Cross-bin metrics have been shown to be more suitable than bin-by-bin metrics for measuring the distance between histograms in various applications. In particular, a visual tracker that minimizes the earth mover's distance (EMD) between the candidate and reference feature histograms has recently been proposed. This tracker was shown to be more robust than the Mean Shift tracker, which employs a bin-by-bin metric. In each frame, the former tracker iteratively shifts the candidate location by one pixel in the direction opposite to the EMD's gradient until no improvement is made. This optimization process involves the clustering of the candidate feature density in feature space, as well as the computation of the EMD between the candidate and reference feature histograms after each shift of the candidate location. In this paper, alternative trackers that employ cross-bin metrics as well, but that are based on Mean Shift (MS) iterations, are derived. The proposed trackers are simpler and faster due to 1) the use of MS-based optimization, which is not restricted to single pixel shifts, 2) abstention from any clustering of feature densities, and 3) abstention from EMD computations in multidimensional spaces.


Three-Dimensional Optical Coherence Tomography Imaging For Glaucoma Associated With Boston Keratoprosthesis Type I and II.
Three-dimensional (3D) spectral domain optical coherence tomography (OCT) volume scans of the optic nerve head (ONH) and the peripapillary area are useful in the management of glaucoma in patients with a type I or II Boston Keratoprosthesis (KPro). The purpose of this study was to report the use of spectral domain OCT in the management of glaucoma in patients with a type I or II Boston KPro. This study is an observational case series. Four consecutive patients with KPro implants were referred for glaucoma evaluation. A comprehensive eye examination was performed which included disc photography, visual field testing, and high-density spectral domain OCT volume scans of the ONH and the peripapillary area. 2D and 3D parameters were calculated using custom-designed segmentation algorithms developed for glaucoma management. Spectral domain OCT parameters provided useful information in the diagnosis and management of 4 KPro patients. OCT parameters which can be used in KPro patients included 2D retinal nerve fiber layer (RNFL) thickness, 3D peripapillary RNFL volume, 3D peripapillary retinal thickness and volume, 3D cup volume, and 3D neuroretinal rim thickness and volume. In 3 of 4 cases where the traditional 2D RNFL thickness scan was limited by artifacts, 3D spectral domain OCT volume scans provided useful quantitative objective measurements of the ONH and peripapillary region. Therefore, 3D parameters derived from high-density volume scans as well as radial scans of the ONH can be used to overcome the limitations and artifacts associated with 2D RNFL thickness scans. Spectral domain OCT volume scans offer the possibility to enhance the evaluation of KPro patients with glaucoma by using both 2D and 3D diagnostic parameters that are easily obtained in a clinic setting.


Repeated imitation makes human vocalizations more word-like.
People have long pondered the evolution of language and the origin of words. Here, we investigate how conventional spoken words might emerge from imitations of environmental sounds. Does the repeated imitation of an environmental sound gradually give rise to more word-like forms? In what ways do these forms resemble the original sounds that motivated them (i.e. exhibit iconicity)? Participants played a version of the children's game 'Telephone'. The first generation of participants imitated recognizable environmental sounds (e.g. glass breaking, water splashing). Subsequent generations imitated the previous generation of imitations for a maximum of eight generations. The results showed that the imitations became more stable and word-like, and later imitations were easier to learn as category labels. At the same time, even after eight generations, both spoken imitations and their written transcriptions could be matched above chance to the category of environmental sound that motivated them. These results show how repeated imitation can create progressively more word-like forms while continuing to retain a resemblance to the original sound that motivated them, and speak to the possible role of human vocal imitation in explaining the origins of at least some spoken words.


Clozapine: a novel antipsychotic with a controversial introduction.
Last year's introduction of clozapine, an atypical antipsychotic, in the United States was the most significant advance in schizoprenia treatment since the advent of chlopromazine in the 1950s. However, clozapine has a disturbingly high incidence of agranulocytosis. The drug manufacturer designed a complex and expensive mandatory blood monitoring system that was dismantled after public pressure earlier this year. There is hope that clozapine will become more widely available to schizoprenic patients.


Bone mineral density as a dose-response predictor for osteoporosis: a propensity score analysis of longitudinal incident study (KCIS no. 39).
Osteoporosis is a global disease burden for aging society. The role of quantitative ultrasound (QUS) in the prediction for osteoporosis in a dose-response manner is hardly addressed. We aimed to show the dose-response of QUS measurement in the prediction for osteoporosis by a community-based study. A prospective cohort study. Participants were recruited between 2000 and 2004. Demographic data and heel QUS measurement were collected at baseline. Diagnosis of osteoporosis was ascertained by the follow-up of this cohort over time. In order to reduce the imbalance of baseline characteristics in the observational study, we applied propensity score by using proportional odds regression analysis to match the quintiles of QUS T-score. A total of 44 957 subjects composed of 17 678 men (39.3%) and 27 279 women (69.7%) were recruited. After adjustments for propensity score, an increase in one unit of QUB T-score led to 7% reduction in the risk for osteoporosis [adjusted odds ratio (OR) = 0.93, 95% confidence interval (CI): 0.89-0.96, P < 0.0001]. Higher quintile of QUS T-score yielded a lower risk of osteoporosis with a gradient relationship [OR: 0.82 (95%CI: 0.72-0.92); OR: 0.81 (95%CI: 0.71-0.91); OR: 0.77 (95%CI: 0.68-0.87) and OR: 0.76 (95%CI: 0.67-0.86)] from the second to highest quintile opposed to first quintile (P < 0.0001). The cumulative incidence of osteoporosis was higher in the lower quintile during follow-up (log-rank test, P < 0.001). QUS is an independent predictor for osteoporosis in a dose-response manner using a large population-based cohort. Due to the lower cost and portability of QUS measurement, the pre-screening for osteoporosis by QUS can be considered in the area with limited resources can be a feasible and alternative method.


[Surgical correction of corneal astigmatism].
Corneal astigmatism can be treated optically or surgically. First, we try to manage the astigmatism with glasses or contact-lenses. If it cannot be managed successfully with optical devices, a surgical approach may be indicated. Corneal wedge resections and relaxing incisions can be effective. The wedge resection is the more difficult procedure with reasonably predictable results. In recent years corneal relaxing incision has gained popularity because it is a simple technique with fairly rapid visual rehabilitation. However, the correction result is quite unpredictable. We try to evaluate the factor that created the astigmatism. The steep and flat corneal meridian are identified with keratoscopy, and the correction is performed at the end of the corneal meridian causing the astigmatism. The technique and results are discussed.


Resumption of menses in anorexia nervosa during a course of family-based treatment.
The resumption of menses (ROM) is considered an important clinical marker in weight restoration for patients with anorexia nervosa (AN). The purpose of this study was to examine ROM in relation to expected body weight (EBW) and psychosocial markers in adolescents with AN. We conducted a retrospective chart review at The University of Chicago Eating Disorders Program from September 2001 to September 2011 (N = 225 females with AN). Eighty-four adolescents (Mean age = 15.1, SD = 2.2) with a DSM-IV diagnosis of AN, presenting with secondary amenorrhea were identified. All participants had received a course of outpatient family-based treatment (FBT), i.e., ~20 sessions over 12 months. Weight and menstrual status were tracked at each therapy session throughout treatment. The primary outcome measures were weight (percent of expected for sex, age and height), and ROM. Mean percent EBW at baseline was 82.0 (SD = 6.5). ROM was reported by 67.9% of participants (57/84), on average at 94.9 (SD = 9.3) percent EBW, and after having completed an average of 13.5 (SD = 10.7) FBT sessions (~70% of standard FBT). Compared to participants without ROM by treatment completion, those with ROM had significantly higher baseline Eating Disorder Examination Global scores (p = .004) as well as Shape Concern (p < .008) and Restraint (p < .002) subscale scores. No other differences were found. Results suggest that ROM occur at weights close to the reference norms for percent EBW, and that high pre-treatment eating disorder psychopathology is associated with ROM. Future research will be important to better understand these differences and their implications for the treatment of adolescents with AN.


[Coronary heart disease risk factors in Croatia and worldwide: results of the Interheart study].
In the past 30 years, an increased cardiovascular disease (CVD) mortality has been observed in both industrialized and transition countries. The latter countries, such as Croatia, are considered to be in the third stage of epidemiological transition, defined as having 35%-65% of total as CVD mortality, predominantly ischemic heart disease and cerebrovascular disease. The CVD epidemic in transition countries is due to increasing rates of hypertension, obesity, smoking and sedentary lifestyle. Among CVD diagnoses, the most important is coronary heart disease (CHD), which varies in incidence among different ethnic groups and countries. Worldwide, it is estimated that nine potentially modifiable risk factors contribute to more than 90% of myocardial infarctions. Approximately 80%-90% of patients with symptomatic CHD and more than 95% of patients who died from CHD had at least one of the four traditional risk factors (smoking, hypertension, hyperlipidemia, and diabetes). Little research has been done to quantify the relationship between the prevalence of CHD risk factors and acute myocardial infarction (AMI) in Croatia. In south Croatia, we expected that specific dietary patterns and lifestyle would have favorable effects on CHD risk. Therefore, we have conducted a case-control study to examine the relationship between several CHD risk factors (smoking, hypertension, hyperlipidemia, diabetes, obesity, alcohol, fruit and vegetable consumption, and physical activity) in AMI patients and persons without previously known CHD in south Croatia. We took part in the INTERHEART study over 4 years (1999-2002). Cases were all eligible patients with first AMI admitted to the Coronary Care Unit, Split University Hospital. Within one month of admission at least one control was recruited and matched to every AMI case by age (+/-5 years) and sex. Exclusion criteria were the same for cases and controls. Structured questionnaires were administered and physical examinations were undertaken in the same manner in cases and controls. Relationship between the risk factors and AMI are presented by odds ratios, estimated by multivariate logistic regression. During the study period 263 cases and 264 controls were enrolled. The proportion of males (74.6%) was threefold that of females. The highest relative difference between the case and control risk factors was noted for current smoking (16.6%; p<0.001), diabetes (10.5%; p<0.001), hypertension (9.0%; p=0.038) and abdominal obesity (18.5%; p<0.001). Ever smoking accounted for 75% higher AMI risk than non-smoking (OR 1.74; p=0.006), while current smoking accounted for a 2.6 time higher risk in comparison to non-smoking (OR 2.58; p<0.001). Diabetes had a threefold risk (OR 2.83; p<0.001). Hypertension accounted for a 70% higher risk (OR 1.68; p=0.007). Abdominal obesity was associated with a significantly increased AMI risk (OR 1.96; p=0.007). The highest apolipoprotein B/apolipoprotein A-1 (ApoB/ApoA-1) tertile accounted for nearly 2.5-fold risk (OR 2.23; p=0.005). Physical activity and daily consumption of fruits and vegetables did not prove to be significant factors in Croatia. Regular consumption of alcohol decreased coronary risk by approximately one third (OR 0.63; p=0.044). The most important AMI risk factor in south Croatia is current smoking, followed by diabetes, abnormal ApoB/ApoA-1 ratio, abdominal obesity, and hypertension. A protective risk factor is alcohol consumption, while physical activity and fruit and vegetable consumption are less important. These results are similar to the global INTERHEART data showing that most of AMI risk could be predicted with nine simple, measurable risk factors worldwide. Protective measures for CHD, including increased daily consumption of fruits and vegetables, moderate physical activity and particularly smoking cessation should be implemented worldwide. In specific regions such as south Croatia, moderate alcohol consumption (mostly red wine) may be included among protective measures due to sociologic and cultural reasons.


Genetic Diversity and Molecular Epidemiology of Mycobacterium tuberculosis in Roraima State, Brazil.
National border areas are special places for the spread of Mycobacterium tuberculosis (MTB). These regions concentrate vulnerable populations and constant population movements. Understanding the dynamics of the transmission of MTB is fundamental to propose control measures and to monitor drug resistance. We conducted a population-based prospective study of tuberculosis (TB) to evaluate molecular characteristics of MTB isolates circulating in Roraima, a state on the border of Venezuela and Guyana. Eighty isolates were genotyped by IS6110-RFLP (restriction fragment length polymorphism), spoligotyping, and 24-locus mycobacterial interspersed repetitive unit-variable number of repeats tandem (MIRU-VNTR). Drug susceptibility tests were performed by using the proportion method and GeneXpert® MTB/RIF (Cepheid, Sunnyvale, CA). Isolates showing a phenotypic resistance profile were submitted to polymerase chain reaction (PCR) and sequencing. Spoligotyping showed 40 distinct patterns with a high prevalence of Latin-American and Mediterranean (LAM), Haarlem (H), and the "ill-defined" T clades. Mycobacterial interspersed repetitive unit -VNTR and IS6110-RFLP showed clustering rates of 21.3% and 30%, respectively. Drug resistance was detected in 11 (15.1%) isolates, and all were found to have primary resistance; among these, six (8.2%) isolates were streptomycin mono-resistant, four (5.4%) isoniazid mono-resistant, and one (1.3%) multidrug resistant. This is the first study on the molecular epidemiology and drug resistance profile of MTB from Roraima. Herein, we describe high diversity of genetic profiles circulating in this region that may be driven by the introduction of new strain types because of large population flow in this region. In summary, our results showed that analyses of these circulating strains can contribute to a better understanding of TB epidemiology in the northern Brazilian border and be useful to establish public health policies on TB prevention.


NAD+ loss, a new player in AhR biology: prevention of thymus atrophy and hepatosteatosis by NAD+ repletion.
Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD) is a carcinogenic and highly toxic industrial byproduct that persists in the environment and produces a pleiotropic toxicity syndrome across vertebrate species that includes wasting, hepatosteatosis, and thymus atrophy. Dioxin toxicities require binding and activation of the aryl hydrocarbon receptor (AhR), a ligand activated transcription factor. However, after nearly 50 years of study, it remains unknown how AhR activation by dioxin produces toxic effects. Here, using the chick embryo close to hatching, a well-accepted model for dioxin toxicity, we identify NAD+ loss through PARP activation as a novel unifying mechanism for diverse effects of dioxin in vivo. We show that NAD+ loss is attributable to increased PARP activity in thymus and liver, as cotreatment with dioxin and the PARP inhibitor PJ34 increased NAD+ levels and prevented both thymus atrophy and hepatosteatosis. Our findings additionally support a role for decreased NAD+ dependent Sirt6 activity in mediating dioxin toxicity following PARP activation. Strikingly, treatment in vivo with the NAD+ repleting agent nicotinamide, a form of vitamin B3, prevented thymus atrophy and hepatosteatosis by dioxin and increased sirtuin activity, providing a therapeutic approach for preventing dioxin toxicities in vivo.


A biochemical study of vitamin B12 deficiency in the baboon.
A dietary deficiency of vitamin B12 was produced in the baboon by use of an artificial diet. Deficiency was further aggravated by feeding propionate or ampicillin, as judged by serum, liver and brain vitamin B12 levels, and urinary excretion of methyl malonic acid. Reduced glutathione levels in the blood and liver increased during deficiency, while ascorbic acid levels were not affected, Brain and liver nucleic acids did not change significantly. Blood pyruvate and serum lipid factors were affected more in the group fed propionate. These results are compared with the work on vitamin B12 deficient rats and pernicious anaemia in humans.


Nitrite reductase from Pseudomonas aeruginosa induces inflammatory cytokines in cultured respiratory cells.
Persistent infection with Pseudomonas aeruginosa increases interleukin-8 (IL-8) levels and causes dense neutrophil infiltrations in the airway of patients with chronic airway diseases. To investigate the role of P. aeruginosa infection in IL-8 production in the airway of these patients, we examined whether cell lysates of P. aeruginosa could cause IL-8 production from human bronchial epithelial cells. Diluted sonicated supernatants of P. aeruginosa (SSPA) with a mucoid or nonmucoid phenotype stimulated human bronchial epithelial (BET-1A) cells to produce IL-8. In this study, we have purified a 59-kDa heat-stable protein with IL-8-inducing activity from the SSPA by sequential ion-exchange chromatography. The N-terminal sequence of this purified protein completely matched a sequence at the N-terminal part of the mature protein of nitrite reductase from P. aeruginosa. In addition, immunoblotting with a polyclonal immunoglobulin G (IgG) against recombinant Pseudomonas nitrite reductase demonstrated a specific binding to the purified protein. Furthermore, the immunoprecipitates of the SSPA with a polyclonal IgG against recombinant nitrite reductase induced a twofold-higher IL-8 production in the BET-1A cell culture than did the immunoprecipitates of the SSPA with a control IgG. These lines of evidence confirmed that Pseudomonas nitrite reductase was responsible for IL-8 production in the BET-1A cells. The purified nitrite reductase induced maximal expression of IL-8 mRNA in the BET-1A cells at 1 to 3 h after stimulation, and the IL-8 mRNA expression declined by 8 h after stimulation. New protein translation was not required for nitrite reductase-mediated IL-8 mRNA expression in the BET-1A cells. Nitrite reductase stimulated the BET-1A cells, as well as human alveolar macrophages, pulmonary fibroblasts, and neutrophils, to produce IL-8. In contrast, nitrite reductase induced significant levels of tumor necrosis factor alpha and IL-1beta protein only in human alveolar macrophages. These data support the notion that nitrite reductase from P. aeruginosa induces the production of inflammatory cytokines by respiratory cells and may contribute to the pathogenesis of chronic airway diseases and persistent P. aeruginosa infection.


Changes in gamma-aminobutyric acid content during beni-koji making.
The changes in the gamma-aminobutyric acid (GABA) content during the making of beni-koji prepared with Monascus pilosus IFO 4520 vs. the difference in the rate of tomo koji (10%, 30%, and 50%) were examined. The increased proportion of tomo koji would increase the GABA production and the productions of GABA peaked on the fifth day and thereafter declined. The glutamate decarboxylase activity during beni-koji making with 50% tomo koji steadily increased after the start of the koji making, reaching its peak on the fifth day. The succinic acid content increased after the sixth day. The mycelial growth was in the stationary phase after the sixth day. Therefore, the GABA content increases with an increase in the proportion of tomo koji. It is presumed that the maximum amount of GABA reached on the fifth day was the cause of the increasing amount of conversion of GABA into succinic acid, in addition to the decline in the GAD activity after the fifth day of koji making.


[The functional organization of the posterior lateral nucleus of the rat hypothalamus].
It has been demonstrated that the posterior lateral thalamic nucleus of the rat receives information from the visual, somatic and auditory systems. Some of the neurons (63%) have a convergent input from these systems, although these neurons exhibit functional specificity with respect to the predominant inhibitory influence of the background activity of one of the sensory systems investigated. The other part of the neurons (37%) receives information only from the visual or somatic system, these neurons exhibiting excitatory phasic reaction to sensory stimuli.


Extending P450 site-of-metabolism models with region-resolution data.
Cytochrome P450s are a family of enzymes responsible for the metabolism of approximately 90% of FDA-approved drugs. Medicinal chemists often want to know which atoms of a molecule-its metabolized sites-are oxidized by Cytochrome P450s in order to modify their metabolism. Consequently, there are several methods that use literature-derived, atom-resolution data to train models that can predict a molecule's sites of metabolism. There is, however, much more data available at a lower resolution, where the exact site of metabolism is not known, but the region of the molecule that is oxidized is known. Until now, no site-of-metabolism models made use of region-resolution data. Here, we describe XenoSite-Region, the first reported method for training site-of-metabolism models with region-resolution data. Our approach uses the Expectation Maximization algorithm to train a site-of-metabolism model. Region-resolution metabolism data was simulated from a large site-of-metabolism dataset, containing 2000 molecules with 3400 metabolized and 30 000 un-metabolized sites and covering nine Cytochrome P450 isozymes. When training on the same molecules (but with only region-level information), we find that this approach yields models almost as accurate as models trained with atom-resolution data. Moreover, we find that atom-resolution trained models are more accurate when also trained with region-resolution data from additional molecules. Our approach, therefore, opens up a way to extend the applicable domain of site-of-metabolism models into larger regions of chemical space. This meets a critical need in drug development by tapping into underutilized data commonly available in most large drug companies. The algorithm, data and a web server are available at http://swami.wustl.edu/xregion.


Centrosome-associated regulators of the G(2)/M checkpoint as targets for cancer therapy.
In eukaryotic cells, control mechanisms have developed that restrain cell-cycle transitions in response to stress. These regulatory pathways are termed cell-cycle checkpoints. The G(2)/M checkpoint prevents cells from entering mitosis when DNA is damaged in order to afford these cells an opportunity to repair the damaged DNA before propagating genetic defects to the daughter cells. If the damage is irreparable, checkpoint signaling might activate pathways that lead to apoptosis. Since alteration of cell-cycle control is a hallmark of tumorigenesis, cell-cycle regulators represent potential targets for therapy. The centrosome has recently come into focus as a critical cellular organelle that integrates G(2)/M checkpoint control and repairs signals in response to DNA damage. A growing number of G(2)/M checkpoint regulators have been found in the centrosome, suggesting that centrosome has an important role in G(2)/M checkpoint function. In this review, we discuss centrosome-associated regulators of the G(2)/M checkpoint, the dysregulation of this checkpoint in cancer, and potential candidate targets for cancer therapy.


Mental disorders and risk of suicide attempt in prisoners.
Mental disorders are overrepresented in prisoners, placing them at an increased risk of suicide. Advancing our understanding of how different mental disorders relate to distinct stages of the suicidal process-the transition from ideation to action-would provide valuable information for clinical risk assessment in this high-risk population. Data were drawn from a representative sample of 1212 adults (1093 men) incarcerated across 13 New Zealand prisons, accounting for 14% of the national prison population. Guided by an ideation-to-action framework, three mutually exclusive groups of participants were compared on the presence of mental disorders assessed by validated DSM-IV diagnostic criteria: prisoners without any suicidal history (controls; n = 778), prisoners who thought about suicide but never made a suicide attempt (ideators; n = 187), and prisoners who experienced suicidal ideation and acted on such thoughts (attempters; n = 247). One-third (34.6%) of participants reported a lifetime history of suicidal ideation, of whom 55.6% attempted suicide at some point (19.2% of all prisoners). Suicidal outcomes in the absence of mental disorders were rare. Whilst each disorder increased the odds of suicidal ideation (OR range 1.73-4.13) and suicide attempt (OR range 1.82-4.05) in the total sample (n = 1212), only a select subset of disorders was associated with suicide attempt among those with suicidal ideation (n = 434). Drug dependence (OR 1.65, 95% CI 1.10-2.48), alcohol dependence (OR 1.89, 95% CI 1.26-2.85), and posttraumatic stress disorder (OR 2.09, 95% CI 1.37-3.17) distinguished attempters from ideators. Consistent with many epidemiological studies in the general population, our data suggest that most mental disorders are best conceptualized as risk factors for suicidal ideation rather than for suicide attempt. Once prisoners consider suicide, other biopsychosocial factors beyond the mere presence of mental disorders may account for the progression from thoughts to acts of suicide.


Autogenous regulation of gene expression.
A new term, autogenous regulation, is used to describe a phenomenon that is not a new discovery but rather is newly appreciated as a mechanism common to a number of systems in both prokaryotic and eukaryotic organisms. In this mechanism the product of a structural gene regulates expression of the operon in which that structural gene resides. In many (perhaps all) cases, the regulatory gene product has several functions, since it may act not only as a regulatory protein but also as an enzyme, structural protein, or antibody, for example. In a few cases, this protein is the multimeric allosteric enzyme that catalyzes the first step of a metabolic pathway, gearing together the two most important mechanisms for controlling the biosynthesis of metabolites in bacterial cells-feedback inhibition and repression. Autogenous regulation may provide a mechanism for amplification of gene expression (84); for severe and prolonged inactivation of gene expression (85); for buffering the response of structural genes to changes in the environment (45, 52); and for maintaining a constant intracellular concentration of a protein, independent of cell size or growth rate (86). Thus, autogenous regulation provides the cell with means for accomplishing a number of different regulatory tasks, each suited to better satisfying the needs of the organism for its survival.


A radiative transfer equation-based image-reconstruction method incorporating boundary conditions for diffuse optical imaging.
Developing reconstruction methods for diffuse optical imaging requires accurate modeling of photon propagation, including boundary conditions arising due to refractive index mismatch as photons propagate from the tissue to air. For this purpose, we developed an analytical Neumann-series radiative transport equation (RTE)-based approach. Each Neumann series term models different scattering, absorption, and boundary-reflection events. The reflection is modeled using the Fresnel equation. We use this approach to design a gradient-descent-based analytical reconstruction algorithm for a three-dimensional (3D) setup of a diffuse optical imaging (DOI) system. The algorithm was implemented for a three-dimensional DOI system consisting of a laser source, cuboidal scattering medium (refractive index > 1), and a pixelated detector at one cuboid face. In simulation experiments, the refractive index of the scattering medium was varied to test the robustness of the reconstruction algorithm over a wide range of refractive index mismatches. The experiments were repeated over multiple noise realizations. Results showed that by using the proposed algorithm, the photon propagation was modeled more accurately. These results demonstrated the importance of modeling boundary conditions in the photon-propagation model.


Synthesis of diverse asparagine linked oligosaccharides and synthesis of sialylglycopeptide on solid phase.
Oligosaccharides are linked to the protein surface and play roles in a number of biological events. Therefore, much attention is being paid to research to investigate the function of the oligosaccharides. In order to investigate the function of oligosaccharides, many synthetic approaches have been examined by synthesizing O-linked or N-linked glycopeptides. Synthesis of O-linked type oligosaccharides is relatively feasible compared to that of N-linked oligosaccharides, because the number of sugar components in the former oligosaccharides is small. In the biosynthesis of oligosaccharides, only N-linked oligosaccharide is reconstructed from the high mannose-type to the hybrid and complex types at the Golgi apparatus. This scientific question, namely, why only N-glycan should change its structure, has been paid much attention and convenient synthesis for both N-glycan and glycopeptide having N-glycans has been examined in order to study the role of N-glycan. In this review, we would introduce recent synthetic developments focusing on the synthesis of N-linked glycopeptides and its analogues.


New therapeutic avenues in ulcerative colitis: thinking out of the box.
Ulcerative colitis (UC) is a chronic idiopathic inflammatory disease of the gastrointestinal tract that affects the mucosal lining of the colon. Recent epidemiological data show that its incidence and prevalence are increasing in many parts of the world, in parallel with altered lifestyles, improved access to health, improved sanitation and industrialisation rates. Current therapeutic strategies for treating UC have only been moderately successful. Despite major recent advances in inflammatory bowel disease therapeutic resources, a considerable proportion of patients are still refractory to conventional treatment. Less than half of all patients achieve long-term remission, many require colectomy, and the disease still has a major impact on patients' lives. Moreover, recent data point to slightly raised mortality. While these outcomes could be partly improved by optimising current therapeutic strategies, they clearly highlight the need to develop new therapies. Currently, a number of promising and innovative therapeutic approaches are being explored, some of which will hopefully survive to reach the clinic. Until such a time arrives, it is important that a better understanding of the clinical particularities of the disease, an improved knowledge of the host-microbiome negative interactions and of the environmental factors beyond disease development is achieved to obtain the final and desired outcome: to provide better treatment and quality of life for patients with this disabling disease.


YqhC regulates transcription of the adjacent Escherichia coli genes yqhD and dkgA that are involved in furfural tolerance.
Previous results have demonstrated that the silencing of adjacent genes encoding NADPH-dependent furfural oxidoreductases (yqhD dkgA) is responsible for increased furfural tolerance in an E. coli strain EMFR9 [Miller et al., Appl Environ Microbiol 75:4315-4323, 2009]. This gene silencing is now reported to result from the spontaneous insertion of an IS10 into the coding region of yqhC, an upstream gene. YqhC shares homology with transcriptional regulators belonging to the AraC/XylS family and was shown to act as a positive regulator of the adjacent operon encoding YqhD and DkgA. Regulation was demonstrated by constructing a chromosomal deletion of yqhC, a firefly luciferase reporter plasmid for yqhC, and by a direct comparison of furfural resistance and NADPH-dependent furfural reductase activity. Closely related bacteria contain yqhC, yqhD, and dkgA orthologs in the same arrangement as in E. coli LY180. Orthologs of yqhC are also present in more distantly related Gram-negative bacteria. Disruption of yqhC offers a useful approach to increase furfural tolerance in bacteria.


Neurogenic hypertension and the secrets of respiration.
Despite recent advances in the knowledge of the neural control of cardiovascular function, the cause of sympathetic overactivity in neurogenic hypertension remains unknown. Studies from our laboratory point out that rats submitted to chronic intermittent hypoxia (CIH), an experimental model of neurogenic hypertension, present changes in the central respiratory network that impact the pattern of sympathetic discharge and the levels of arterial pressure. In addition to the fine coordination of respiratory muscle contraction and relaxation, which is essential for O2 and CO2 pulmonary exchanges, neurons of the respiratory network are connected precisely to the neurons controlling the sympathetic activity in the brain stem. This respiratory-sympathetic neuronal interaction provides adjustments in the sympathetic outflow to the heart and vasculature during each respiratory phase according to the metabolic demands. Herein, we report that CIH-induced sympathetic over activity and mild hypertension are associated with increased frequency discharge of ventral medullary presympathetic neurons. We also describe that their increased frequency discharge is dependent on synaptic inputs, mostly from neurons of the brain stem respiratory network, rather than changes in their intrinsic electrophysiological properties. In perspective, we are taking into consideration the possibility that changes in the central respiratory rhythm/pattern generator contribute to increased sympathetic outflow and the development of neurogenic hypertension. Our experimental evidence provides support for the hypothesis that changes in the coupling of respiratory and sympathetic networks might be one of the unrevealed secrets of neurogenic hypertension in rats.


Neural Substrate for Metacognitive Accuracy of Tactile Working Memory.
The human prefrontal cortex (PFC) has been shown to be important for metacognition, the capacity to monitor and control one's own cognitive processes. Here we dissected the neural architecture of somatosensory metacognition using navigated single-pulse transcranial magnetic stimulation (TMS) to modulate tactile working memory (WM) processing. We asked subjects to perform tactile WM tasks and to give a confidence rating for their performance after each trial. We circumvented the challenge of interindividual variability in functional brain anatomy by applying TMS to two PFC areas that, according to tractography, were neurally connected with the primary somatosensory cortex (S1): one area in the superior frontal gyrus (SFG), another in the middle frontal gyrus (MFG). These two PFC locations and a control cortical area were stimulated during both spatial and temporal tactile WM tasks. We found that tractography-guided TMS of the SFG area selectively enhanced metacognitive accuracy of tactile temporal, but not spatial WM. Stimulation of the MFG area that was also neurally connected with the S1 had no such effect on metacognitive accuracy of either the temporal or spatial tactile WM. Our findings provide causal evidence that the PFC contains distinct neuroanatomical substrates for introspective accuracy of tactile WM.


Effect of O6-benzylguanine analogues on sensitivity of human tumor cells to the cytotoxic effects of alkylating agents.
The effect of O6-benzylguanine, O6-(p-chlorobenzyl)guanine, and O6-(p-methylbenzyl)guanine on the sensitivity of various human tumor cell lines to alkylating agents is evaluated. The sensitivity of human colon tumor cells, HT29, to the chloroethylating agents, 1,3-bis(2-chloroethyl)-1-nitrosourea, 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea, 2-chloroethyl(methylsulfonyl) methanesulfonate (clomesone), and chlorozotocin was increased by pretreatment for 2 h with 25 microM of each analogue. O6-Benzylguanine was slightly more effective as a sensitizer in HT29 cells than the p-chlorobenzyl and p-methylbenzyl analogues. However, all analogues sensitized SF767 glioma cells to the cytotoxic effects of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea, 1,3-bis(2-chloroethyl)-1-nitrosourea, and clomesone to the same degree. Both cell lines were sensitized to the methylating agents streptozotocin and 5-(3-methyl-1-triazeno)imidazole-4-carboxamide, the active intermediate of 5-(3,3-dimethyl-1-triazenyl)imidazole-4-carboxamide, by pretreatment with 10 microM O6-benzylguanine for 2 h. The number of Raji cells surviving 50 microM clomesone decreased 3-fold upon pretreatment for 2 h with 1 microM O6-benzylguanine. The degree of enhancement was dependent on the amount of alkyltransferase protein present in cell lines. For example, HT29 cells (alkyltransferase activity, 381 fmol/mg protein) exhibited a greater degree of enhancement when treated with O6-benzylguanine than SF767 (77 fmol/mg protein) and M19-MEL melanoma (36 fmol/mg protein) cells. There was no enhancement observed in mer- cell lines, U251 (less than 2 fmol/mg protein), and BE (3 fmol/mg protein), or with alkylating agents which did not produce a cytotoxic lesion at the O6 position of guanine in DNA such as cisplatin or 4-hydroperoxycyclophosphamide. Our studies suggest that O6-benzylguanine analogues may have utility in mer+ tumors as an adjuvant to a variety of alkylating agents which produce a toxic lesion at the O6 position of guanine.


MYH9 is associated with nondiabetic end-stage renal disease in African Americans.
As end-stage renal disease (ESRD) has a four times higher incidence in African Americans compared to European Americans, we hypothesized that susceptibility alleles for ESRD have a higher frequency in the West African than the European gene pool. We carried out a genome-wide admixture scan in 1,372 ESRD cases and 806 controls and found a highly significant association between excess African ancestry and nondiabetic ESRD (lod score = 5.70) but not diabetic ESRD (lod = 0.47) on chromosome 22q12. Each copy of the European ancestral allele conferred a relative risk of 0.50 (95% CI = 0.39-0.63) compared to African ancestry. Multiple common SNPs (allele frequencies ranging from 0.2 to 0.6) in the gene encoding nonmuscle myosin heavy chain type II isoform A (MYH9) were associated with two to four times greater risk of nondiabetic ESRD and accounted for a large proportion of the excess risk of ESRD observed in African compared to European Americans.


Dependence of internal friction on folding mechanism.
An outstanding challenge in protein folding is understanding the origin of "internal friction" in folding dynamics, experimentally identified from the dependence of folding rates on solvent viscosity. A possible origin suggested by simulation is the crossing of local torsion barriers. However, it was unclear why internal friction varied from protein to protein or for different folding barriers of the same protein. Using all-atom simulations with variable solvent viscosity, in conjunction with transition-path sampling to obtain reaction rates and analysis via Markov state models, we are able to determine the internal friction in the folding of several peptides and miniproteins. In agreement with experiment, we find that the folding events with greatest internal friction are those that mainly involve helix formation, while hairpin formation exhibits little or no evidence of friction. Via a careful analysis of folding transition paths, we show that internal friction arises when torsion angle changes are an important part of the folding mechanism near the folding free energy barrier. These results suggest an explanation for the variation of internal friction effects from protein to protein and across the energy landscape of the same protein.


Cyclic Strain Affects Macrophage Cytokine Secretion and Extracellular Matrix Turnover in Electrospun Scaffolds.
Electrospun elastomeric scaffolds are being used for a variety of in situ tissue engineering applications, in which biomechanical loads play a dominant in vivo role, such as cardiovascular replacements (e.g., heart valve and blood vessel prostheses) and pelvic floor reconstruction. The findings of this study underline that immunomodulatory scaffolds for biomechanically loaded applications should be mechanically tailored, for example, in terms of stiffness and compliance, to support a desirable proregenerative macrophage phenotype. Moreover, this research contributes to the general understanding of pathophysiological macrophage phenotypes in cyclically strained tissues (e.g., atherosclerotic plaques), and their role in tissue regeneration and degeneration.


Adverse Drug Reaction Discovery from Electronic Health Records with Deep Neural Networks.
Adverse drug reactions (ADRs) are detrimental and unexpected clinical incidents caused by drug intake. The increasing availability of massive quantities of longitudinal event data such as electronic health records (EHRs) has redefined ADR discovery as a big data analytics problem, where data-hungry deep neural networks are especially suitable because of the abundance of the data. To this end, we introduce neural self-controlled case series (NSCCS), a deep learning framework for ADR discovery from EHRs. NSCCS rigorously follows a self-controlled case series design to adjust implicitly and efficiently for individual heterogeneity. In this way, NSCCS is robust to time-invariant confounding issues and thus more capable of identifying associations that reflect the underlying mechanism between various types of drugs and adverse conditions. We apply NSCCS to a large-scale, real-world EHR dataset and empirically demonstrate its superior performance with comprehensive experiments on a benchmark ADR discovery task.


Ten caveats of learning analytics in health professions education: A consumer's perspective.
A group of 22 medical educators from different European countries, gathered in a meeting in Utrecht in July 2019, discussed the topic of learning analytics (LA) in an open conversation and addressed its definition, its purposes and potential risks for learners and teachers. LA was seen as a significant advance with important potential to improve education, but the group felt that potential drawbacks of using LA may yet be under-exposed in the literature. After transcription and interpretation of the discussion's conclusions, a document was drafted and fed back to the group in two rounds to arrive at a series of 10 caveats educators should be aware of when developing and using LA, including too much standardized learning, with undue consequences of over-efficiency and pressure on learners and teachers, and a decrease of the variety of 'valid' learning resources. Learning analytics may misalign with eventual clinical performance and can run the risk of privacy breaches and inescapability of documented failures. These consequences may not happen, but the authors, on behalf of the full group of educators, felt it worth to signal these caveats from a consumers' perspective.


[Extracorporeal photopheresis in treatment of chronic graft versus host disease].
Extracorporeal photopheresis (ECP) is an immunomodulatory therapy which has been used in the treatment of chronic GVHD (cGVHD). ECP involves separation of the mononuclear cells with leukapheresis, followed by ex vivo administration of 8-methoxypsoralen and UV-A radiation and reinfusion to the patient. Aim of the study was to evaluate clinical and immunomodulatory effect of ECP procedures in patients with cGVHD. We analyzed 341 ECP procedures performed in 7 patients with cGVHD; median ECP per patient was 37 (range 13-131). All patients suffered from skin changes in combination with impaired joint mobility and symptoms of oral disease. ECP procedures were performed for two consecutive days: in initial phase weekly, followed by every two weeks and than monthly according to clinical response. Median of ECP treatment duration was 10 months (range 2-58). The effect of ECP in patients with cGVHD with skin andjoint involvement was mostly beneficial: 6 patients experienced either improvement or stabilization in skin changes and joint mobility. In 2 patients who suffered from oral disease, the total recovery was observed. Clinical response was typically delayed until 2 to 3 months, and reduction in glucocorticoid dose was observed. Adverse reactions were observed in 4.9% procedures. In patients who responded to ECP treatment, CD4+/CD8+ ratio and number of NK cells were normalized. ECP proved to be an efficient and safe procedure that may be recommended for patients with cGVHD who do not respond to conventional therapy.


Partial Proton Transfer in the Gas Phase: A Spectroscopic and Computational Analysis of the Trifluoroacetic Acid - Trimethylamine Complex.
The 1:1 complex formed from trifluoroacetic acid (TFA) and trimethylamine (TMA) has been observed in the gas phase by rotational spectroscopy and further investigated by DFT and MP2 methods. Spectra of both the parent form and the -OD isotopologue have been obtained. The complex is structurally similar to a hydrogen bonded system, with the O-H bond directed toward the nitrogen of the TMA. However, both the spectroscopic and computational results indicate that it is intermediate between a hydrogen bonded complex and a proton-transferred ion pair. Two metrics are used to assess the degree of proton transfer from the acid to the base. The first is based on experimental 14N nuclear quadrupole coupling constants. Specifically, the component of the 14N nuclear quadrupole coupling tensor along the c-inertial axis of the complex, χcc, is 31% of the way between that of free TMA (no proton transfer) and that of TMAH+ (complete proton transfer). A second metric, adapted from that of Kurnig and Scheiner [Int. J. Quantum Chem. Quantum Biol. Symp. 1987, 14, 47-56], is based on calculated O-H and H-N distances and corroborates this description. These results indicate that the degree of proton transfer in TFA-TMA is very similar to that in the TMA complex of HNO3, which has been previously studied and for which the proton affinity of the conjugate anion (NO3-) is almost identical to that of CF3COO-. While the solid salt, TMAH+·CF3COO-, is an ionic plastic above 307 K and exhibits free rotation of the ions, no such motion is observed in the cold 1:1 gas phase adduct.


Measurement of yunaconitine and crassicauline A in small-volume blood serum samples by LC-MS/MS: tracing of aconite poisoning in clinical diagnosis.
Aconite poisoning is one of the most serious types of herb-related medical emergencies. In Hong Kong, many if not most of these poisoning cases are due to confusion in herbal species; that is, the wrong herbs are used in prescriptions. Such human errors, while inevitable perhaps, can be serious, and sometimes fatal. The chemical components responsible for aconite poisoning are yunaconitine and crassicauline A. In the present study, a rapid and sensitive method for the screening and quantification of yunaconitine and crassicauline A in human serum, using LC-MS/MS, was developed and validated. Methyllycaconitine was chosen as the internal standard. The limit of detection (LOD) of yunaconitine and crassicauline A were found to be 0.022 and 0.021 ng/mL, respectively. The limit of quantification (LOQ) was 0.1 ng/mL for both yunaconitine and crassicauline A. The recovery of yunaconitine and crassicauline A ranged from 78.6% to 84.9% and 78.3% to 87.2%, respectively. The matrix effect of yunaconitine and crassicauline A ranged from 110.0% to 130.4% and 121.2 to 130.0%, respectively. Both yunaconitine and crassicauline A were stable in serum for at least 3 months at -20 °C, and the extracts were stable for at least 7 days. For clinical applications, serum samples of two patients confirmed to have had aconite herbs poisoning in 2008 were quantified using the developed method. The result showed that this method can be utilized in clinical routine applications. This screening method expedites the diagnosis in cases of suspected aconite poisoning, thus enabling doctors to treat the condition more quickly and effectively.


Operando analysis reveals potential-driven in situ formation of single-Fe-atom electrocatalysts for green production of ammonia.
As a sustainable approach for N2 fixation, electrocatalytic N2 reduction reaction (N2RR) to produce ammonia (NH3) is highly desirable with a precise understanding to the structure-activity relationship of electrocatalysts. Here, firstly, we obtain a novel carbon-supported oxygen-coordinated single-Fe-atom catalyst for highly efficient production of ammonia from electrocatalytic N2RR. Based on such new type of N2RR electrocatalyst, by combining operando X-ray absorption spectra (XAS) with density function theory calculation, we reveal significantly that the as-prepared active coordination structure undergoes a potential-driven two-step restructuring, firstly from FeSAO4(OH)1a to FeSAO4(OH)1a'(OH)1b with the adsorption of another -OH on FeSA at open-circuit potential (OCP) of 0.58 VRHE, and subsequently restructuring from FeSAO4(OH)1a'(OH)1b to FeSAO3(OH)1a″ due to the breaking of one Fe-O bond and the dissociation of one -OH at working potentials for final electrocatalytic process of N2RR, thus revealing the first potential-induced in situ formation of the real electrocatalytic active sites to boost the conversion of N2 to NH3. Moreover, the key intermediate of Fe-NNHx was detected experimentally by both operando XAS and in situ attenuated total reflection-surface-enhanced infrared absorption spectra (ATR-SEIRAS), indicating the alternating mechanism followed by N2RR on such catalyst. The results indicate the necessity of considering the potential-induced restructuring of the active sites on all kinds of electrocatalysts for such as highly efficient ammonia production from N2RR. It also paves a new way for a precise understanding to the structure-activity relationship of a catalyst and helps the design of highly efficient catalysts.


Epidemiology of Echinococcus granulosus infection in the central Peruvian Andes.
The prevalence of human, canine, and ovine echinococcosis was determined in an endemic area of the Peruvian Andes where control programmes have not been operational since 1980. Prevalence of infection in humans was determined using portable ultrasound, chest X-rays, and an enzyme-linked immunoelectrotransfer blot (EITB) assay. Canine and ovine echinococcal prevalence was determined by microscopic stool examinations following arecoline purging for tapeworm detection and by examination of the viscera from slaughtered livestock animals, respectively. The prevalence among 407 humans surveyed was 9.1%. The frequency of disease in the liver, lung, and in both organs was 3.4%, 2.0%, and 0.2%, respectively. Portable ultrasound or portable chest X-ray has shown that, compared to adults, children under 11 years had significantly higher seropositive rates without evidence of hydatid disease (P < 0.05). Among the 104 dogs inspected for echinococcus after arecoline purging, 33 (32%) were positive for adult tapeworms. Among the 117 sheep slaughtered at the local abattoir, 102 (87%) had hydatid cysts. The prevalence of human hydatidosis in this endemic area of Peru is one of the highest in the world and nearly five times higher than previously reported in 1980. An increase in echinococcosis prevalence may result after premature cessation of control programmes.


Imaging Evaluation of Nonacute Shoulder Pain.
A variety of radiographic views have been described in the literature for the initial imaging of nonacute shoulder pain. Multiple articles have also discussed the best next imaging test if radiographs do not show the diagnosis, especially the use of unenhanced MRI, MR arthrography, CT arthrography, and ultrasound. The purpose of this article is to examine the evidence for the most helpful radiographic views and the best imaging test after radiographs for different clinical presentations of chronic shoulder pain. The recommended radiographic views and best next imaging test after the radiograph depends on the clinical diagnosis. Ultrasound is generally preferred over MRI for evaluating chronic rotator cuff pain, whereas MR arthrography, preferably with abducted and externally rotated images, is most accurate for imaging chronic symptoms from a suspected labral tear or instability.


Peripheral mitochondrial DNA, telomere length and DNA methylation as predictors of live birth in in vitro fertilization cycles.
To evaluate whether telomere length (TL), mitochondrial-DNA (mt-DNA) or epigenetic age estimators based on DNA methylation (DNAm) pattern could be considered reliable predictors of in-vitro-fertilization (IVF) success in terms of live birth rate. Prospective cohort study. Infertility Unit of the Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico. 181 women aged 37-39 years who underwent IVF at a single-centre between January 2017 and December 2018. On the day of recruitment, blood samples were collected, and genomic DNA was isolated from white blood cells. TL, mt-DNA and DNAm assessment was performed using quantitative real-time polymerase chain reaction (qPCR). Biological age (DNAm age) was computed as the algorithm based on methylation pattern of five genes. Epigenetic age acceleration was estimated from the residuals of the linear model of epigenetic age regressed on chronological age. Long Interspersed Nuclear Elements (LINE)-1 methylation pattern was used as a surrogate for global DNA methylation. This study investigated whether peripheral TL, mt-DNA and DNAm could predict live birth in IVF cycles. TL, mt-DNA and LINE-1 methylation were not associated with IVF success. Conversely, DNAm age resulted significantly lower in women who had a live birth compared to women who did not (36.1 ± 4.2 and 37.3 ± 3.3 years, respectively, p = 0.04). For DNAm age, odds ratio (OR) for live birth per year of age was 0.90 (95%CI: 0.82-0.99, p = 0.036) after adjusting for FSH and antral follicle count (AFC) and 0.90 (95%CI: 0.82-0.99, p = 0.028) after adjusting also for number of oocytes retrieved. A significant association also emerged for epigenetic age acceleration after adjustments (OR = 0.91, 95%CI: 0.83-1.00, p = 0.048). DNAm age is associated with IVF success but the magnitude of this association is insufficient to claim a clinical use. However, our findings are promising and warrant further investigation. Assessment of biological age using different epigenetic clocks or focusing on different tissues may reveal new predictors of IVF success.


Effects of serial percutaneous application of carbon dioxide in intermittent claudication: results of a controlled trial.
In a prospective, controlled clinical trial, serial application of carbon-dioxide-enriched water was compared with fresh water. Twenty-four patients with peripheral arterial occlusive disease (stable claudication) were randomly allocated to one of two serial intervention groups, lower extremities immersed in either fresh water or in CO2-enriched water (1000 mg CO2/kg) water under standardized conditions (temperature, 33 degrees C; depth, 40 cm; immersion time, 30 min; five times a week over 4 weeks). The serial application of carbon-dioxide-enriched water increased arterial peak flow (reactive hyperemia), transcutaneous oxygen tension (basal value and half-recovery-time), and pain-free walking distance. The serial fresh water application did not change these values. The authors conclude that serial carbon dioxide application can be clinically effective in patients with arterial obstructions in the lower extremities.


Looking forward to 20/20: a focus on the epidemiology of eye diseases.
The encouraging scenario of international efforts to eliminate preventable and avoidable blindness is the legacy of public health ophthalmology in the 20th century. With active programs currently in place or beginning for the major cause of blindness in childhood and two of the leading infectious causes of blindness, it is natural that research in eye disease will shift even more heavily toward the leading causes of blindness in the older ages. The age-related eye diseases will rapidly become the most common causes of blindness and visual loss and, with the exception of cataract, are the more difficult to identify, diagnose, and treat. The human misery and social cost of blindness, especially in the countries that can ill afford it, are profound. To combat this problem, epidemiologic research in ophthalmology should look toward the following major areas: 1. the identification and testing of better screening modalities to determine early changes possibly amenable to preventive strategies. This includes detection of vitamin A deficiency as well. 2. the creation of uniform definitions for diseases, particularly glaucoma and early AMD, which have relevance for epidemiologic research into risk factors. 3. increased multidisciplinary research, working with investigators skilled in molecular genetics, biologic markers for age-related diseases, and those interested in new imaging and vision-testing techniques. 4. ongoing work in clinical trials of new approaches to prevent or delay the onset of vision loss from eye disease, including future vaccines for chlamydia and onchocerciasis. The major public health issue of blindness prevention will not disappear in the next century but only shift emphasis to different causes if the current programs achieve the success that is hoped. Future epidemiologic research will continue to require a concerted, sustained, and multidisciplinary effort in order to contribute to the vision research agenda in the next century.