bluuebunny/crossref_metadata_2025 huggingface dataset:
Reducing signs of aging and increasing lifespan by drug synergy
Authors: Huang Xinhe , Liu Jun , Withers Bradley R. , Samide Aaron J. , Leggas Markos , Dickson Robert C.
Published: April 2013

Disease incidence rises rapidly with age and increases both human suffering and economic hardship while shortening life. Advances in understanding the signaling pathways and cellular processes that influence aging support the possibility of reducing the incidence of age‐related diseases and increasing lifespan by pharmacological intervention. Here, we demonstrate a novel pharmacological strategy that both reduces signs of aging in the budding yeast Saccharomyces cerevisiae and generates a synergistic increase in lifespan. By combining a low dose of rapamycin, to reduce activity of the target of rapamycin complex 1 (TORC1) protein kinase, and myriocin, to reduce sphingolipid synthesis, we show enhancement of autophagy, genomic stability, mitochondrial function, and AMP kinase pathway activity. These processes are controlled by evolutionarily conserved signal transduction pathways that are vital for maintaining a healthy state and promoting a long life. Thus, our data show that it ought to be possible to find pharmacological approaches to generate a synergistic reduction in the incidence of human age‐related diseases to improve health quality in the elderly and enhance lifespan.
DSM‐5 criteria for depression with mixed features: a farewell to mixed depression
Authors: Koukopoulos A. , Sani G.
Published: April 2013

Objective

To review the DSM‐5 proposed criteria for mixed depression in light of robust and consistent historical and scientific evidence.

Method

An extensive historical search, a systematic review of the papers used by DSM‐5 as reference papers, and a PubMed search were performed.

Results

As Hippocrates, depressive mixed states have been described as conditions of intense psychic suffering, consisting of depressed mood, inner tension, restlessness, and aimless psychomotor agitation. In DSM‐5, new criteria are proposed for a mixed features specifier, as part of depression either in major depressive disorder (MDD) or bipolar disorder. Those criteria require, as diagnostically specific, manic/hypomanic symptoms that are the least common kinds of symptoms that actually arise in depressive mixed states. The DSM‐5 proposal is based, almost entirely, on a speculative wish to avoid ‘overlapping’ manic and depressive symptoms. Mixed states are, in fact, nothing but overlapping manic and depressive symptoms.

Conclusion

In this article, we review the psychopathology and research on mixed depressive states, and try to demonstrate that the DSM‐5 proposal has weak scientific basis and does not identify a large number of mixed depressive states. This may be harmful because of the different treatment required by these conditions.
Long‐term biometric optic components of diode laser‐treated threshold retinopathy of prematurity at 9 years of age
Authors: Yang Chang‐Sue , Wang An‐Guor , Shih Yung‐Feng , Hsu Wen‐Ming
Published: April 2013

Purpose:  To assess quantitatively the biometric optic components and its relationship with refractive status in preterm school children with diode laser‐treated threshold retinopathy of prematurity (ROP).

Methods:  A prospective, cross‐sectional study in which ultrasound biometric measurement of optic components and cycloplegic refraction were performed on 24 consecutive preterm children with diode laser‐treated threshold ROP at the age of 9 years. The study results were compared with 1021 age‐matched full‐term control children from a national survey.

Results:  The eyes with laser‐treated ROP showed statistically significantly thicker lens (3.94 versus 3.39 mm), steeper vertical corneal curvature (7.47 versus 7.67 mm) and shallower anterior chamber depth (ACD) (2.91 versus 3.58 mm) than age‐matched full‐term controls, but no difference in axial length (23.32 versus 23.24 mm). The laser‐treated eyes had a mean spherical equivalent (SE) of −4.49 D compared with mean SE of −0.44 D in controls. Of 46 eyes studied, 93% of eyes were myopic and 28.3% with high myopia (<−6.0 D) compared with the 32% prevalence of myopia in controls. In preterm children, younger gestational age tended to correlate with shallower ACD (r = 0.352) and thicker lens (r = −0.298); lower birth weight tended to correlate with shallower ACD (r = 0.372) and steeper cornea (r = 0.360).

Conclusions:  There were higher prevalence and greater magnitude of myopia in preterm children. The significantly thicker lens, steeper corneal curvature and shallower anterior chamber depth are the major factors contributing to the development of myopia in preterm school children with laser‐treated threshold ROP.
Identification of drug candidate against prostate cancer from the aspect of somatic cell reprogramming
Authors: Kosaka Takeo , Nagamatsu Go , Saito Shigeru , Oya Mototsugu , Suda Toshio , Horimoto Katsuhisa
Published: April 2013

Considering the similarities between the transcriptional programming involved in cancer progression and somatic cell reprogramming, we tried to identify drugs that would be effective against malignant cancers. We used the early transposon Oct4 and Sox2 enhancer (EOS) system to select human prostate cancer (PCA) cells expressing high levels of OCT4. Patients with metastatic castration‐resistant PCA that does not respond to treatment with docetaxel have few therapeutic options. The OCT4‐expressing PCA cells selected using the EOS system showed increased tumorigenicity and high resistance to docetaxel, both in vitro and in vivo. By using their gene expression data, expression signature‐based prediction for compound candidates identified an antiviral drug, ribavirin, as a conversion modulator from drug resistance to sensitivity. Treatment of PCA cells with ribavirin decreased their resistance against treatment with docetaxel. This indicated that ribavirin reversed the gene expression, including that of humoral factors, in the OCT4‐expressing PCA cells selected using the EOS system. Thereby, ribavirin increased the efficacy of docetaxel for cancer cells. We propose a novel cell reprogramming approach, named drug efficacy reprogramming, as a new model for identifying candidate antitumor drugs.
Design, Synthesis, and Biological Evaluation of Catecholamine Vehicle for Studying Dopaminergic System
Authors: Singh Pooja , Kumar Vikas , Aggarwal Swati , Tiwari Anjani K. , Chuttani Krishna , Pratap Ramendra , Mishra Anil K.
Published: April 2013

Catecholamine mimetic EDTA‐bis(tyramide) was synthesized and characterized by various spectroscopic techniques (NMR, mass spectroscopy) and λem 310 nm for the excitation at 270 nm. Molecular docking studies were performed with human serum albumin (PDB 1E78), showing binding pattern with amino acid residues Arg218, Arg222, and Lys444, identifies the ligand‐human serum albumin interaction for the transportation affinity of the ligand at the specific site of the target. Subsequently, binding study with human serum albumin at λex = 350 nm found to be 5.847 × 104 m−1 shows effective quenching effect. Additionally, to go more insight, acetylcholinesterase binding affinity was investigated, which shows 90% binding affinity for the 10 mm concentration. IC50 value was found 18.60 μm for MAO‐B inhibition. Finally, EDTA‐bis(tyramide) labeled with 99mTc to investigate its in vivo radiopharmaceutical efficiency having 97% binding affinity with 98% radiochemical purity. In vivo studies were carried out for 99mTc‐EDTA‐bis(tyramide) included blood kinetics showed a quick wash out from the circulation via renal route, and biodistribution revealed that maximum %ID/g was found in kidney at 1 h, and its scintigraphy image shows 3.96% brain uptake with respect to whole body.
Discovery of a New Bioactive Molecule for Neuroblastoma
Authors: Leitão Andrei , Schramm Alexander , Eggert Angelika
Published: April 2013

Neuroblastoma, a common pediatric malignancy of neural crest origin, is unique in its wide spectrum of clinical and biological behavior, ranging from spontaneous regression or differentiation to rapid progression and metastasis. Overexpression of neurotrophin receptors of the tyrosine kinase (Trk) family has been identified as a major prognostic and biological factor for this disease. Novel molecules were selected using cheminformatics tools (structure‐based virtual screening and ligand‐based virtual screening) and screened in cell‐based assays to modulate Trk receptor activity. One compound (C390‐0031) had a potent antiproliferative activity in dose–response studies using neuroblastoma cell lines. The molecular effects of this molecule were further characterized by using cell cycle and Western blot analysis. Interestingly, despite the presence of the anchoring fragment to the Trk kinase domain composing its structure, this molecule does not inhibit TrkA like lestaurtinib, constituting a new chemical with a yet unknown mechanism of action.
Physicochemical and Biological Characterization of Pep‐1/Elastin Complexes
Authors: Ahmad Nasrollahi Saman , Taghibiglou Changiz , Fouladdel Shamileh , Dinarvand Rasoul , Moosavi Movahedi Ali A. , Azizi Ebrahim , Farboud Effat S.
Published: April 2013

Transdermal drug delivery of proteins is challenging because the skin acts as a natural and protective barrier. Several techniques including using the cell‐penetrating peptides have been studied to increase the penetration of therapeutic proteins into and through the skin. Cell‐penetrating peptides facilitate and improve the transduction of large and hydrophilic cargo molecules through plasma membrane. We have recently reported an efficient skin delivery of elastin protein in complex with a cell‐penetrating peptide called Pep‐1. As the biophysical characteristics of cell‐penetrating peptide/protein complexes have been linked with their biological responses, in this study, we investigated biophysical properties of Pep‐1/elastin complexes (ratio 10:1) stored in three temperatures (−20 °C, 4 °C and 25 °C) by photon correlation spectroscopy, circular dichroism and isothermal denaturation. We also evaluated the ability of transduction of this complex into cells and skin tissue using both fluorescence microscopy and Kodak In‐Vivo FX Pro Imaging System.
The potential of sarcospan in adhesion complex replacement therapeutics for the treatment of muscular dystrophy
Authors: Marshall Jamie L. , Kwok Yukwah , McMorran Brian J. , Baum Linda G. , Crosbie‐Watson Rachelle H.
Published: April 2013

Three adhesion complexes span the sarcolemma and facilitate critical connections between the extracellular matrix and the actin cytoskeleton: the dystrophin– and utrophin–glycoprotein complexes and α7β1 integrin. Loss of individual protein components results in a loss of the entire protein complex and muscular dystrophy. Muscular dystrophy is a progressive, lethal wasting disease characterized by repetitive cycles of myofiber degeneration and regeneration. Protein‐replacement therapy offers a promising approach for the treatment of muscular dystrophy. Recently, we demonstrated that sarcospan facilitates protein–protein interactions amongst the adhesion complexes and is an important potential therapeutic target. Here, we review current protein‐replacement strategies, discuss the potential benefits of sarcospan expression, and identify important experiments that must be addressed for sarcospan to move to the clinic.
Structural diversity of calmodulin binding to its target sites
Authors: Tidow Henning , Nissen Poul
Published: April 2013

Calmodulin (CaM) is a ubiquitous, highly conserved, eukaryotic protein that binds to and regulates a number of diverse target proteins involved in different functions such as metabolism, muscle contraction, apoptosis, memory, inflammation and the immune response. In this minireview, we analyze the large number of CaM‐complex structures deposited in the Protein Data Bank (i.e. crystal and nuclear magnetic resonance structures) to gain insight into the structural diversity of CaM‐binding sites and mechanisms, such as those for CaM‐activated protein kinases and phosphatases, voltage‐gated Ca2+‐channels and the plasma membrane Ca2+‐ATPase.
The adaptive immune response does not influence hantavirus disease or persistence in the Syrian hamster
Authors: Prescott Joseph , Safronetz David , Haddock Elaine , Robertson Shelly , Scott Dana , Feldmann Heinz
Published: April 2013

Pathogenic New World hantaviruses cause severe disease in humans characterized by a vascular leak syndrome, leading to pulmonary oedema and respiratory distress with case fatality rates approaching 40%. Hantaviruses infect microvascular endothelial cells without conspicuous cytopathic effects, indicating that destruction of the endothelium is not a mechanism of disease. In humans, high levels of inflammatory cytokines are present in the lungs of patients that succumb to infection. This, along with other observations, suggests that disease has an immunopathogenic component. Currently the only animal model available to study hantavirus disease is the Syrian hamster, where infection with Andes virus (ANDV), the primary agent of disease in South America, results in disease that closely mimics that seen in humans. Conversely, inoculation of hamsters with a passaged Sin Nombre virus (SNV), the virus responsible for most cases of disease in North America, results in persistent infection with high levels of viral replication. We found that ANDV elicited a stronger innate immune response, whereas SNV elicited a more robust adaptive response in the lung. Additionally, ANDV infection resulted in significant changes in the blood lymphocyte populations. To determine whether the adaptive immune response influences infection outcome, we depleted hamsters of CD4+ and CD8+ T cells before infection with hantaviruses. Depletion resulted in inhibition of virus‐specific antibody responses, although the pathogenesis and replication of these viruses were unaltered. These data show that neither hantavirus replication, nor pathogenesis caused by these viruses, is influenced by the adaptive immune response in the Syrian hamster.
Numerical chromosome disorders in the common marmoset (Callithrix jacchus) – comparison between two captive colonies
Authors: Delimitreva S. , Wedi E. , Bakker J. , Tkachenko O.Y. , Nikolova V. , Nayudu P.L.
Published: April 2013

Background

Chromosomal analyses were performed for marmosets from two colonies – Deutsches Primatenzentrum (DPZ) and Biomedical Primate Research Centre (BPRC). Chlorine‐based disinfectants are used in DPZ; no chemical disinfection is applied in BPRC.

Methods

The rates of chromosomal non‐disjunction, polyploidy and endoreduplication were investigated after G‐banding.

Results

For DPZ monkeys, the mean rates of non‐disjunction were 7.6% for bone marrow and 11.3% for lymphocytes. The polyploidy level was 2.5% in bone marrow and 0.8% in blood. Frequency of endoreduplication in bone marrow and in leucocytes was 0.5% and 0.8%, respectively. For BPRC, the rate of non‐disjunction in leucocytes (1.3%) was significantly lower than that for DPZ; the polyploidy rate (0.2%) in blood was lower than that in DPZ; endoreduplication was not observed.

Conclusion

The levels of chromosomal disorders are elevated for DPZ colony. We suggest that the increased rate of chromosomal disorders in DPZ marmosets can be related to the chemical disinfection of their environment.
Molecular analysis of the site for 2‐arachidonylglycerol (2‐AG) on the β2 subunit of GABAA receptors
Authors: Baur Roland , Kielar Marie , Richter Lars , Ernst Margot , Ecker Gerhard F. , Sigel Erwin
Published: April 2013

2‐arachidonyl glycerol (2‐AG) allosterically potentiates GABAA receptors via a binding site located in transmembrane segment M4 of the β2 subunit. Two amino acid residues have been described that are essential for this effect. With the aim to further describe this potential drug target, we performed a cysteine scanning of the entire M4 and part of M3. All four residues in M4 affecting the potentiation here and the two already identified residues locate to the same side of the α‐helix. This side is exposed to M3, where further residues were identified. From the fact that the important residues span > 18 Å, we conclude that the hydrophobic tail of the bound 2‐AG molecule must be near linear and that the site mainly locates to the inner leaflet but stretches far into the membrane. The influence of the structure of the head group of the ligand molecule on the activity of the molecule was also investigated. We present a model of 2‐AG docked to the GABAA receptor.
Harnessing the power of yeast to unravel the molecular basis of neurodegeneration
Authors: Tenreiro Sandra , Munder Matthias C. , Alberti Simon , Outeiro Tiago F.
Published: April 2013

Several neurodegenerative diseases, such as Parkinson's disease (PD), Alzheimer's disease (AD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), or prion diseases, are known for their intimate association with protein misfolding and aggregation. These disorders are characterized by the loss of specific neuronal populations in the brain and are highly associated with aging, suggesting a decline in proteostasis capacity may contribute to pathogenesis. Nevertheless, the precise molecular mechanisms that lead to the selective demise of neurons remain poorly understood. As a consequence, appropriate therapeutic approaches and effective treatments are largely lacking. The development of cellular and animal models that faithfully reproduce central aspects of neurodegeneration has been crucial for advancing our understanding of these diseases. Approaches involving the sequential use of different model systems, starting with simpler cellular models and ending with validation in more complex animal models, resulted in the discovery of promising therapeutic targets and small molecules with therapeutic potential. Within this framework, the simple and well‐characterized eukaryote Saccharomyces cerevisiae, also known as budding yeast, is being increasingly used to study the molecular basis of several neurodegenerative disorders. Yeast provides an unprecedented toolbox for the dissection of complex biological processes and pathways. Here, we summarize how yeast models are adding to our current understanding of several neurodegenerative disorders.
Calcium/calmodulin‐dependent serine protein kinase (CASK) is a new intracellular modulator of P2X3 receptors
Authors: Gnanasekaran Aswini , Sundukova Mayya , Hullugundi Swathi , Birsa Nicol , Bianchini Giorgio , Hsueh Yi‐Ping , Nistri Andrea , Fabbretti Elsa
Published: April 2013

ATP‐gated P2X3 receptors of sensory ganglion neurons are important transducers of painful stimuli and are modulated by extracellular algogenic substances, via changes in the receptor phosphorylation state. The present study investigated the role of calcium/calmodulin‐dependent serine protein kinase (CASK) in interacting and controlling P2X3 receptor expression and function in mouse trigeminal ganglia. Most ganglion neurons in situ or in culture co‐expressed P2X3 and CASK. CASK was immunoprecipitated with P2X3 receptors from trigeminal ganglia and from P2X3/CASK‐cotransfected human embryonic kidney (HEK) cells. Recombinant P2X3/CASK expression in HEK cells increased serine phosphorylation of P2X3 receptors, typically associated with receptor upregulation. CASK deletion mutants also enhanced P2X3 subunit expression. After silencing CASK, cell surface P2X3 receptor expression was decreased, which is consistent with depressed P2X3 currents. The reduction in P2X3 expression levels was reversed by the proteasomal inhibitor MG‐132. Moreover, neuronal CASK/P2X3 interaction was up‐regulated by nerve growth factor (NGF) signaling and down‐regulated by P2X3 agonist‐induced desensitization. These data suggest a novel interaction between CASK and P2X3 receptors with positive outcome for receptor stability and function. As CASK‐mediated control of P2X3 receptors was dependent on the receptor activation state, CASK represents an intracellular gateway to regulate purinergic nociceptive signaling.
Altered stress responsiveness and hypothalamic‐pituitary‐adrenal axis function in male rat offspring of socially isolated parents
Authors: Pisu Maria Giuseppina , Garau Anna , Olla Pierluigi , Biggio Francesca , Utzeri Cinzia , Dore Riccardo , Serra Mariangela
Published: April 2013

Social isolation in male rats at weaning results in reduced basal levels of the neuroactive steroid 3α,5α‐tetrahydroprogesterone (3α,5α‐TH PROG) in the brain and plasma as well as increased anxiety‐like behavior. We now show that socially isolated female rats also manifest a reduced basal cerebrocortical concentration of 3α,5α‐TH PROG as well as an anxiety‐like profile in the elevated plus‐maze and Vogel conflict tests compared with group‐housed controls. In contrast, despite the fact that they were raised under normal conditions, adult male offspring of male and female rats subjected to social isolation before mating exhibited an increased basal cerebrocortical level of 3α,5α‐TH PROG but no difference in emotional reactivity compared with the offspring of group‐housed parents. These animals also showed an increased basal activity of the hypothalamic‐pituitary‐adrenal axis as well as reduced abundance of corticotropin‐releasing factor in the hypothalamus and of corticotropin‐releasing factor receptor type 1 in the pituitary. Moreover, negative feedback regulation of hypothalamic‐pituitary‐adrenal axis activity by glucocorticoid was enhanced in association with up‐regulation of glucocorticoid receptor expression in the hippocampus. There was also attenuation of corticosterone release induced by foot‐shock stress in the offspring of socially isolated parents. The increase in the brain concentration of 3α,5α‐TH PROG induced by acute stress was also blunted in these animals. Our results thus show that a stressful experience before mating can influence neuroendocrine signaling in the next generation.
24(S)‐hydroxycholesterol is actively eliminated from neuronal cells by ABCA1
Authors: Matsuda Akihiro , Nagao Kohjiro , Matsuo Michinori , Kioka Noriyuki , Ueda Kazumitsu
Published: April 2013

High cholesterol turnover catalyzed by cholesterol 24‐hydroxylase is essential for neural functions, especially learning. Because 24(S)‐hydroxycholesterol (24‐OHC), produced by 24‐hydroxylase, induces apoptosis of neuronal cells, it is vital to eliminate it rapidly from cells. Here, using differentiated SH‐SY5Y neuron‐like cells as a model, we examined whether 24‐OHC is actively eliminated via transporters induced by its accumulation. The expression of ABCA1 and ABCG1 was induced by 24‐OHC, as well as TO901317 and retinoic acid, which are ligands of the nuclear receptors liver X receptor/retinoid X receptor (LXR/RXR). When the expression of ABCA1 and ABCG1 was induced, 24‐OHC efflux was stimulated in the presence of high‐density lipoprotein (HDL), whereas apolipoprotein A‐I was not an efficient acceptor. The efflux was suppressed by the addition of siRNA against ABCA1, but not by ABCG1 siRNA. To confirm the role of each transporter, we analyzed human embryonic kidney 293 cells stably expressing human ABCA1 or ABCG1; we clearly observed 24‐OHC efflux in the presence of HDL, whereas efflux in the presence of apolipoprotein A‐I was marginal. Furthermore, the treatment of primary cerebral neurons with LXR/RXR ligands suppressed the toxicity of 24‐OHC. These results suggest that ABCA1 actively eliminates 24‐OHC in the presence of HDL as a lipid acceptor and protects neuronal cells.
Associations between EYA1 single‐nucleotide polymorphisms and non‐syndromic orofacial clefts in Western Han Chinese
Authors: Zeng Ni , Wu Jun , Zhu Wen‐Chao , Ma Li , Jia Zhong‐Lin , Shi Bing
Published: April 2013

Background

Non‐syndromic orofacial clefts (NSOC) are the most common craniofacial birth defects and are considered as complex and multifactorial diseases involving the interplay of genetic predisposition and environmental exposures. Recent studies demonstrated that EYA1, a member of eye absent gene family, might contribute to the development of orofacial clefts. This study aimed to evaluate the association between EYA1 gene polymorphism and NSOC in western Chinese population.

Method

Two single‐nucleotide polymorphisms, rs13260349 and rs2380716, were investigated among 199 case–parent trios, and the linkage disequilibrium (LD) test, transmission disequilibrium test, family based association test, and haplotype analysis were performed.

Results

Weak evidence of LD was found between these markers (D' = 0.573, r2 = 0.218); allel C and genotype C/C at rs13260349 showed an overtransmission in CL/P; haplotype T‐T (rs13260349‐rs2380716) displayed an association with NSOC and CL/P.

Conclusions

The results showed evidence of weak association between the two SNPs of EYA1 (rs13260349 and rs2380716) and NSOC.
Influence of clinically significant portal hypertension on survival after hepatic resection for hepatocellular carcinoma in cirrhotic patients
Authors: Giannini Edoardo G. , Savarino Vincenzo , Farinati Fabio , Ciccarese Francesca , Rapaccini Gianludovico , Marco Mariella Di , Benvegnù Luisa , Zoli Marco , Borzio Franco , Caturelli Eugenio , Chiaramonte Maria , Trevisani Franco ,   The Italian Liver Cancer (ITA.LI.CA) group
Published: April 2013

Background

The role of clinically significant portal hypertension on the prognosis of cirrhotic patients undergoing hepatic resection for hepatocellular carcinoma (HCC) is debated.

Aims

In this study, our aim was to assess the role of clinically significant portal hypertension after hepatic resection for HCC in patients with cirrhosis.

Methods

We assessed the prognostic role of the presence of clinically significant portal hypertension (oesophageal/gastric varices/portal hypertensive gastropathy or a platelet count <100 × 109/L associated with splenomegaly) in 152 patients with compensated cirrhosis who underwent hepatic resection for HCC at the Italian Liver Cancer centres. Survival rates were assessed in the whole series, in the subgroup of Child‐Pugh score 5 patients with uninodular HCC ≤5 cm, and in Child‐Pugh score 5 patients with uninodular HCC ≤2 cm and normal bilirubin.

Results

Median survival was similar in patients with and without clinically significant portal hypertension (79 vs 77 months, P = 0.686). Child‐Pugh score 5 was the only variable significantly associated with survival by Cox multiple regression (P = 0.007). In Child‐Pugh score 5 patients with single HCC ≤5 cm or in those with single HCC ≤2 cm and normal bilirubin, there was no survival difference between patients with and without clinically significant portal hypertension (median survival: 94 vs 78 months, P = 0.121 and >100 vs 86 months, P = 0.742).

Conclusions

Presence of clinically significant portal hypertension has no influence on survival of patients with well‐compensated cirrhosis undergoing hepatic resection for HCC.
The inpatient economic and mortality impact of hepatocellular carcinoma from 2005 to 2009: analysis of the US nationwide inpatient sample
Authors: Mishra Alita , Otgonsuren Munkhzul , Venkatesan Chapy , Afendy Mariam , Erario Madeline , Younossi Zobair M.
Published: April 2013

Background

Hepatocellular carcinoma (HCC) is an important complication of cirrhosis. Our aim was to assess the inpatient economic and mortality of HCC in the USA

Methods

Five cycles of Nationwide Inpatient Sample (NIS) conducted from 2005 to 2009 were used. Demographics, inpatient mortality, severity of illness, payer type, length of stay (LoS) and charges were available. Changes and associated factors related to inpatient HCC were assessed using simple linear regression. Odds ratios and 95% CIs for hospital mortality were analysed using log‐linked regression model. To estimate the sampling variances for complex survey data, we used Taylor series approach. SAS® v.9.3 was used for statistical analysis.

Results

From 2005 to 2009, 32,697,993 inpatient cases were reported to NIS. During these 5 years, primary diagnosis of HCC increased from 4401 (2005), 4170 (2006), 5065 (2007), 6540 (2008) to 6364 (2009). HCC as any diagnosis increased from 68 per 100 000 discharges (2005) to 99 per 100 000 (2009). However, inpatient mortality associated with HCC decreased from 12% (2005) to 10% (2009) (P < 0.046) and LoS remained stable. However, median inflation‐adjusted charges at the time of discharge increased from $29,466 per case (2005) to $31,656 per case (2009). Total national HCC charges rose from $1.0 billion (2005) to $2.0 billion (2009). In multivariate analysis, hospital characteristic was independently associated with decreasing in‐hospital mortality (all P < 0.05). Liver transplantation for HCC was the main contributor to high inpatient charges. Longer LoS and other procedures also contributed to higher inpatient charges.

Conclusions

There is an increase in the number of inpatient cases of HCC. Although inpatient mortality is decreasing and the LoS is stable, the inpatient charges associated with HCC continue to increase.
Association of age‐dependent liver injury and fibrosis with immune cell populations
Authors: Collins Bradley H. , Holzknecht Zoie E. , Lynn Kellie A. , Sempowski Gregory D. , Smith Catherine C. , Liu Songling , Parker William , Rockey Don C.
Published: April 2013

Background & Aims

The liver's response to injury is fibrosis, and when chronic, cirrhosis. Age is a critical factor impacting many immune‐mediated processes, potentially including the liver's wounding response to injury.

Methods

The effects of age on acute and chronic liver injury were evaluated using a carbon tetrachloride model in mice. Lymphocyte and macrophage populations were assessed by flow cytometry and immunohistochemical analysis.

Results

Acute liver injury was greater in 18‐month‐old (old) mice than in 9‐month‐old (middle‐aged) mice as judged by changes in aminotransferases. Similarly, livers of 18‐month‐old mice had a significantly greater fibrogenic response to injury than did livers of 9‐month‐old mice after chronic injury (assessed by col1α1 mRNA expression, morphometric analysis and hydroxyproline measurement). Interestingly, livers from young mice (6 weeks old) also exhibited an increase in fibrogenesis compared to 9‐month‐old mice, albeit not to the same degree as in old mice. Consistent with a role for macrophages in fibrogenesis, the number of liver macrophages in young and 9‐month‐old mice increased, while in chronically injured livers of 18‐month‐old mice, the number of macrophages was reduced, and was less than in the livers of young and 9‐month‐old injured livers.

Conclusions

Our data indicate that the fibrogenic response to injury varies substantially with age, and moreover that macrophage recruitment and dynamics may be an important component in differential age‐associated fibrotic disease.
Depositional processes on oceanic island shelves – Evidence from storm‐generated Neogene deposits from the mid‐North Atlantic
Authors: Meireles Ricardo P. , Quartau Rui , Ramalho Ricardo S. , Rebelo Ana C. , Madeira José , Zanon Vittorio , Ávila Sérgio P.
Published: April 2013

Oceanic islands – such as the Azores in the mid‐North Atlantic – are periodically exposed to large storms that often remobilize and transport marine sediments along coastlines, and into deeper environments. Such disruptive events create deposits – denominated tempestites – whose characteristics reflect the highly dynamic environment in which they were formed. Tempestites from oceanic islands, however, are seldom described in the literature and little is known about storm‐related sediment dynamics affecting oceanic island shelves. Therefore, the geological record of tempestite deposits at oceanic islands can provide invaluable information on the processes of sediment remobilization, transport and deposition taking place on insular shelves during and after major storms. In Santa Maria Island (Azores), a sequence of Neogene tempestite deposits was incorporated in the island edifice by the ongoing volcanic activity (thus preserved) and later exposed through uplift and erosion. Because it was overlain by a contemporary coastal lava delta, the water depth at the time of deposition could be inferred, constituting an excellent case‐study to gain insight on the still enigmatic processes of insular shelf deposition. Sedimentological, palaeontological, petrographic and palaeo‐water depth information allowed the reconstruction of the depositional environment of these sediments. The sequence typifies the characteristics of a tempestite (or successive tempestites) formed at ca 50 m depth, in a steep, energetic open insular shelf, and with evidence for massive sediment remobilization from the nearshore to the middle or outer shelf. The authors claim that cross‐shelf transport induced by storm events is the main process of sediment deposition acting on steep and narrow shelves subjected to high‐energetic environments, such as the insular shelves of open‐sea volcanic islands.
Neonatal stress augments the hypoxic chemoreflex of adult male rats by increasing AMPA receptor‐mediated modulation
Authors: Gulemetova Roumiana , Drolet Guy , Kinkead Richard
Published: April 2013

New Findings

What is the central question of this study?

Early life exposure to stress augments the hypoxic chemoreflex of adult male rats. This effect predisposes to respiratory instability during sleep. We used pharmacological and neuroanatomical approaches to determine whether a change in glutamatergic neurotransmission contributes to this abnormal respiratory phenotype.

What is the main finding and its importance?

We initially showed that neonatal stress augments sensitivity to an AMPA/kainate receptor antagonist. Results from autoradiography experiments support this observation because they demonstrate that neonatal stress increases expression of AMPA receptors in key regions that regulate breathing. Region‐specific changes in brain‐derived neurotrophic factor expression may contribute to these changes in glutamatergic neurotransmission. These results bring new insight into the pathophysiology of sleep‐disordered breathing.

Neonatal stress disrupts the developmental trajectory of homeostatic systems. Adult (8‐ to 10‐week‐old) male rats exposed to maternal separation (a form of neonatal stress) display several traits reported in patients suffering from sleep‐disordered breathing, including an augmented hypoxic chemoreflex. To understand the mechanisms behind this effect, we tested the hypothesis that neonatal stress augments glutamatergic neurotransmission in three regions involved in respiratory regulation, namely the nucleus of the solitary tract, the paraventricular nucleus of the hypothalamus and the phrenic motor nucleus. Maternal separation was performed for 3 h day−1 from postnatal day 3 to 12. Control pups were undisturbed. Adult rats were instrumented for intracerebroventricular injection of the AMPA/kainate receptor antagonist CNQX (0–4.3 μm). Using plethysmography, ventilatory activity was measured at rest in awake animals during normoxia (fractional inspired O2= 0.21) and during acute hypoxia (fractional inspired O2= 0.12; 20 min). Following vehicle injection, the hypoxic ventilatory response of stressed rats was 35% greater than that of controls. Microinjection of CNQX attenuated the hypoxic ventilatory response, but the effect observed in stressed rats was greater than that in control animals. Autoradiography experiments showed that neonatal stress augments expression of AMPA receptors within the paraventricular nucleus of the hypothalamus and the phrenic motor nucleus. Quantification of brain‐derived neurotrophic factor showed that neonatal stress augments brain‐derived neurotrophic factor expression only within the paraventricular nucleus. We conclude that neonatal stress augments the hypoxic chemoreflex by increasing the efficacy of glutamatergic synaptic inputs projecting onto key respiratory structures, especially the paraventricular nucleus of the hypothalamus. These data provide new insight into the aetiology of sleep‐disordered breathing.
A novel approach to the Langendorff technique: preparation of isolated cardiomyocytes and myocardial samples from the same rat heart
Authors: MacDougall David A. , Calaghan Sarah
Published: April 2013

New Findings

What is the central question of this study?

The isolation of cardiomyocytes and the homogenization of myocardium are commonly performed on separate rodent hearts. We asked whether it might be viable to derive cells as well as whole muscle (homogenized or cryopreserved) from one organ.

What is the main finding and its importance?

We provide examples of six diverse experiments that can be performed using cells and tissue samples generated by a modified Langendorff technique. The volume and power of resultant data are increased, and the method may be especially beneficial in models of cardiovascular disease.

Researchers in biomedical sciences must continually re‐evaluate their investment in experiments using laboratory animals. Our group is interested in various signalling pathways that underlie physiological and pathophysiological functioning of the mammalian heart. Two important resources for this type of work are isolated cardiomyocytes and homogenized or preserved sections of whole myocardium. In order to improve our experimental approach ethically, we devised an adaptation of the Langendorff whole‐heart retrograde perfusion technique that allows the isolation of adult rat ventricular cardiomyocytes and processing/storage of myocardium from the same heart. This could result in a 50% reduction in the number of animals required for certain experiments. We believe that a generalized adoption of this method would represent a meaningful reduction of animal use in our field of research and, furthermore, strengthen data sets by permitting correlation between myocyte function and various parameters of myocardial biochemistry/structure in individual hearts. This approach is of particular relevance for studies of cardiac pathology, given the cost and time involved in generating experimental disease models.
Physiological Changes in Rhizobia after Growth in Peat Extract May Be Related to Improved Desiccation Tolerance
Authors: Casteriano Andrea , Wilkes Meredith A. , Deaker Rosalind
Published: April 2013

Improved survival of peat-cultured rhizobia compared to survival of liquid-cultured cells has been attributed to cellular adaptations during solid-state fermentation in moist peat. We have observed improved desiccation tolerance of
Rhizobium leguminosarum
bv. trifolii TA1 and
Bradyrhizobium japonicum
CB1809 after aerobic growth in water extracts of peat. Survival of TA1 grown in crude peat extract was 18-fold greater than that of cells grown in a defined liquid medium but was diminished when cells were grown in different-sized colloidal fractions of peat extract. Survival of CB1809 was generally better when grown in crude peat extract than in the control but was not statistically significant (
P
> 0.05) and was strongly dependent on peat extract concentration. Accumulation of intracellular trehalose by both TA1 and CB1809 was higher after growth in peat extract than in the defined medium control. Cells grown in water extracts of peat exhibit morphological changes similar to those observed after growth in moist peat. Electron microscopy revealed thickened plasma membranes, with an electron-dense material occupying the periplasmic space in both TA1 and CB1809. Growth in peat extract also resulted in changes to polypeptide expression in both strains, and peptide analysis by liquid chromatography-mass spectrometry indicated increased expression of stress response proteins. Our results suggest that increased capacity for desiccation tolerance in rhizobia is multifactorial, involving the accumulation of trehalose together with increased expression of proteins involved in protection of the cell envelope, repair of DNA damage, oxidative stress responses, and maintenance of stability and integrity of proteins.
Extracellular Location of Thermobifida fusca Cutinase Expressed in Escherichia coli BL21(DE3) without Mediation of a Signal Peptide
Authors: Su Lingqia , Woodard Ronald W. , Chen Jian , Wu Jing
Published: April 2013

Cutinase is a multifunctional esterase with potential industrial applications. In the present study, a truncated version of the extracellular
Thermobifida fusca
cutinase without a signal peptide (referred to as cutinase
NS
) was heterologously expressed in
Escherichia coli
BL21(DE3). The results showed that the majority of the cutinase activity was located in the culture medium. In a 3-liter fermentor, the cutinase activity in the culture medium reached 1,063.5 U/ml (2,380.8 mg/liter), and the productivity was 40.9 U/ml/h. Biochemical characterization of the purified cutinase
NS
showed that it has enzymatic properties similar to those of the wild-type enzyme. In addition,
E. coli
cells producing inactive cutinase
NS
S130A were constructed, and it was found that the majority of the inactive enzyme was located in the cytoplasm. Furthermore,
T. fusca
cutinase was confirmed to have hydrolytic activity toward phospholipids, an important component of the cell membrane. Compared to the cells expressing the inactive cutinase
NS
S130A, the cells expressing cutinase
NS
showed increased membrane permeability and irregular morphology. Based on these results, a hypothesis of “cell leakage induced by the limited phospholipid hydrolysis of cutinase
NS
” was proposed to explain the underlying mechanism for the extracellular release of cutinase
NS
.
Electrochemical Investigation of a Microbial Solar Cell Reveals a Nonphotosynthetic Biocathode Catalyst
Authors: Strycharz-Glaven Sarah M. , Glaven Richard H. , Wang Zheng , Zhou Jing , Vora Gary J. , Tender Leonard M.
Published: April 2013

Microbial solar cells (MSCs) are microbial fuel cells (MFCs) that generate their own oxidant and/or fuel through photosynthetic reactions. Here, we present electrochemical analyses and biofilm 16S rRNA gene profiling of biocathodes of sediment/seawater-based MSCs inoculated from the biocathode of a previously described sediment/seawater-based MSC. Electrochemical analyses indicate that for these second-generation MSC biocathodes, catalytic activity diminishes over time if illumination is provided during growth, whereas it remains relatively stable if growth occurs in the dark. For both illuminated and dark MSC biocathodes, cyclic voltammetry reveals a catalytic-current–potential dependency consistent with heterogeneous electron transfer mediated by an insoluble microbial redox cofactor, which was conserved following enrichment of the dark MSC biocathode using a three-electrode configuration. 16S rRNA gene profiling showed
Gammaproteobacteria
, most closely related to
Marinobacter
spp., predominated in the enriched biocathode. The enriched biocathode biofilm is easily cultured on graphite cathodes, forms a multimicrobe-thick biofilm (up to 8.2 μm), and does not lose catalytic activity after exchanges of the reactor medium. Moreover, the consortium can be grown on cathodes with only inorganic carbon provided as the carbon source, which may be exploited for proposed bioelectrochemical systems for electrosynthesis of organic carbon from carbon dioxide. These results support a scheme where two distinct communities of organisms develop within MSC biocathodes: one that is photosynthetically active and one that catalyzes reduction of O
2
by the cathode, where the former partially inhibits the latter. The relationship between the two communities must be further explored to fully realize the potential for MSC applications.
Assessment of Virulence Factors Characteristic of Human Escherichia coli Pathotypes and Antimicrobial Resistance in O157:H7 and Non-O157:H7 Isolates from Livestock in Spain
Authors: Cabal A. , Gómez-Barrero S. , Porrero C. , Bárcena C. , López G. , Cantón R. , Gortázar C. , Domínguez L. , Álvarez J.
Published: April 2013

The distribution of virulence factors (VFs) typical of diarrheagenic
Escherichia coli
and the antimicrobial resistance (AMR) profiles were assessed in 780 isolates from healthy pigs, broilers, and cattle from Spain. VF distribution was broader than expected, although at low prevalence for most genes, with AMR being linked mainly to host species.
Repression of Antibiotic Downregulator WblA by AdpA in Streptomyces coelicolor
Authors: Lee Han-Na , Kim Jin-Su , Kim Pil , Lee Heung-Shick , Kim Eung-Soo
Published: April 2013

The upstream region of antibiotic downregulatory
wblA
in
Streptomyces coelicolor
was found to contain AdpA binding motifs. A key morphological regulator, AdpA was shown to specifically bind these motifs by electrophoretic mobility shift assay. An
adpA
disruption mutant exhibited increased
wblA
transcription, suggesting that AdpA negatively regulates
wblA
transcription in
S. coelicolor
.
Genetic Diversity of the Flagellin Genes of Clostridium botulinum Groups I and II
Authors: Woudstra Cedric , Lambert Dominic , Anniballi Fabrizio , De Medici Dario , Austin John , Fach Patrick
Published: April 2013

Botulinum neurotoxins (BoNTs) are produced by phenotypically and genetically different
Clostridium
species, including
Clostridium botulinum
and some strains of
Clostridium baratii
(serotype F) and
Clostridium butyricum
(serotype E). BoNT-producing clostridia responsible for human botulism encompass strains of group I (secreting proteases, producing toxin serotype A, B, or F, and growing optimally at 37°C) and group II (nonproteolytic, producing toxin serotype E, B, or F, and growing optimally at 30°C). Here we report the development of real-time PCR assays for genotyping
C. botulinum
strains of groups I and II based on flaVR (variable region sequence of
flaA
) sequences and the
flaB
gene. Real-time PCR typing of regions flaVR1 to flaVR10 and
flaB
was optimized and validated with 62 historical and Canadian
C. botulinum
strains that had been previously typed. Analysis of 210 isolates of European origin allowed the identification of four new
C. botulinum
flaVR types (flaVR11 to flaVR14) and one new flaVR type specific to
C. butyricum
type E (flaVR15). The genetic diversity of the flaVR among
C. botulinum
strains investigated in the present study reveals the clustering of flaVR types into 5 major subgroups. Subgroups 1, 3, and 4 contain proteolytic
Clostridium botulinum
, subgroup 2 is made up of nonproteolytic
C. botulinum
only, and subgroup 5 is specific to
C. butyricum
type E. The genetic variability of the flagellin genes carried by
C. botulinum
and the possible association of flaVR types with certain geographical areas make gene profiling of flaVR and
flaB
promising in molecular surveillance and epidemiology of
C. botulinum
.
A Double Staining Method Using SYTOX Green and Calcofluor White for Studying Fungal Parasites of Phytoplankton
Authors: Gerphagnon Mélanie , Latour Delphine , Colombet Jonathan , Sime-Ngando Télesphore
Published: April 2013

We propose a double staining method based on the combination of two fluorochromes, calcofluor white (CFW; specific chitinous fluorochrome) and SYTOX green (nucleic acid stain), coupled to epifluorescence microscopy for counting, identifying, and investigating the fecundity of parasitic fungi of phytoplankton and the putative relationships established between hosts and their chytrid parasites. The method was applied to freshwater samples collected over two successive years during the terminal period of autumnal cyanobacterial blooms in a eutrophic lake. The study focused on the uncultured host-parasite couple
Anabaena macrospora
(cyanobacterium) and
Rhizosiphon akinetum
(Chytridiomycota). Our results showed that up to 36.6% of cyanobacterial akinetes could be parasitized by fungi. Simultaneously, we directly investigated the zoosporic content inside the sporangia and found that both the host size and intensity of infection conditioned the final size and hence fecundity of the chytrids. We found that relationships linking host size, final parasite size, and chytrid fecundity were conserved from year to year and seemed to be host-chytrid couple specific. We concluded that our double staining method was a valid procedure for improving our knowledge of uncultured freshwater phytoplankton-chytrid couples and so of the quantitative ecology of chytrids in freshwater ecosystems.