Lipopolysaccharide triggers nuclear import of Lpcat1 to regulate inducible gene expression in lung epithelia

AIM:To report that Lpcat1 plays an important role in regulating lipopolysaccharide (LPS) inducible gene tran-scription. METHODS:Gene expression in Murine Lung Epithelial MLE-12 cells with LPS treatment or Haemophilus influenza and Escherichia coli infection was analyzed by employing quantitative Reverse Transcription Polymerase Chain Reaction techniques. Nucleofection was used to deliver Lenti-viral system to express or knock down Lpcat1 in MLE cells. Subcellular protein fractionation and Western blotting were utilized to study Lpcat1 nuclear relocation. RESULTS:Lpcat1 translocates into the nucleus from thecytoplasm in murine lung epithelia (MLE) after LPS treatment. Haemophilus influenza and Escherichia coli , two LPS-containing pathogens that cause pneumonia, triggered Lpcat1 nuclear translocation from the cytoplasm. The LPS inducible gene expression profile was determined by quantitative reverse transcription polymerase chain reaction after silencing Lpcat1 or overexpression of the enzyme in MLE cells. We detected that 17 out of a total 38 screened genes were upregulated, 14 genes were suppressed, and 7 genes remained unchanged in LPS treated cells in comparison to controls. Knockdown of Lpcat1 by shRNA dramatically changed the spectrum of the LPS inducible gene transcription, as 18 genes out of 38 genes were upregulated, of which 20 genes were suppressed or unchanged. Notably, in Lpcat1 overex-pressed cells, 25 genes out of 38 genes were reduced in the setting of LPS treatment.CONCLUSION:These observations suggest that Lpcat1 relocates into the nucleus in response to bacterial infection to differentially regulate gene transcriptional repression.

Establishing the presence or absence of chronic kidney disease:Uses and limitations of formulas estimating the glomerular filtration rate

The development of formulas estimating glomerular filtration rate(eG FR) from serum creatinine and cystatin C and accounting for certain variables affecting the production rate of these biomarkers, including ethnicity, gender and age, has led to the current scheme of diagnosing and staging chronic kidney disease(CKD),which is based on e GFR values and albuminuria.This scheme has been applied extensively in various populations and has led to the current estimates of prevalence of CKD. In addition, this scheme is applied in clinical studies evaluating the risks of CKD and the efficacy of various interventions directed towards improving its course. Disagreements between creatinine-based and cystatin-based e GFR values and between e GFR values and measured GFR have been reported in various cohorts. These disagreements are the consequence of variations in the rate of production and in factors, other than GFR, affecting the rate of removal of creatinine and cystatin C. The disagreements create limitations for all e GFR formulas developed so far. The main limitations are low sensitivity in detecting early CKD in several subjects, e.g., those with hyperfiltration, and poor prediction of the course of CKD. Research efforts in CKD are currently directed towards identification of biomarkers that are better indices of GFR than the current biomarkers and,particularly, biomarkers of early renal tissue injury.

Effects of the neurotrophic factor artemin on sensory afferent development and sensitivity

Artemin is a neuronal survival and differentiation factor in the glial cell line-derived neurotrophic factor family.Its receptor GFRα3 is expressed by a subpopulation of nociceptor type sensory neurons in the dorsal root and trigeminal ganglia(DRG and TG).These neurons co-express the heat,capsaicin and proton-sensitive channel TRPV1 and the cold and chemical-sensitive channel TRPA1.To further investigate the effects of artemin on sensory neurons,we isolated transgenic mice(ARTN-OE mice) that overexpress artemin in keratinocytes of the skin and tongue.Enhanced levels of artemin led to a 20% increase in the total number of DRG neurons and increases in the level of mRNA encoding TRPV1 and TRPA1.Calcium imaging showed that isolated sensory neurons from ARTN-OE mice were hypersensitive to the TRPV1 agonist capsaicin and the TRPA1 agonist mustard oil.Behavioral testing of ARTN-OE mice also showed an increased sensitivity to heat,cold,capsaicin and mustard oil stimuli applied either to the skin or in the drinking water.Sensory neurons from wildtype mice also exhibited potentiated capsaicin responses following artemin addition to the media.In addition,injection of artemin into hindpaw skin produced transient thermal hyperalgesia.These findings indicate that artemin can modulate sensory function and that this regulation may occur through changes in channel gene expression.Because artemin mRNA expression is up-regulated in inflamed tissue and following nerve injury,it may have a significant role in cellular changes that underlie pain associated with pathological conditions.Manipulation of artemin expression may therefore offer a new pain treatment strategy.

The Motivations and Experiences of Young Women in a Microbicide Trial in the USA and Puerto Rico

Young women are an important target group in microbicide research, yet little is known about why they participate and stay in microbicide trials. Our study examined motivations for participating in a Phase I microbicide trial among 61 women ages 18 - 24 years in the continental USA and Puerto Rico. We also examined their perspectives on study participation. Participants underwent a semi-structured in-depth interview in which they were asked about factors that motivated enrollment and their experiences while participating. They also completed a Web-based Computer Assisted Self Interview in which they were asked to rate study burden (1 = low to 4 = high). Factors that motivated enrollment were altruism (29%), compensation (17%), a combination of altruism and compensation (37%) and free medical exams (17%). Factors that encouraged participants to stay in the study were study staff (95%), confirmation of good health (41%), and the opportunity to learn about their bodies (17%). Mean ratings of study burden ranged from 1.83 (having to travel to site) to 2.41 (colposcopy), indicating that participants were not highly bothered by visits or procedures. Although Phase I trials require invasive procedures, participants were not highly bothered by them and recognized them as necessary. Good relationships with staff and clear information about how procedures contribute to study goals may encourage participants to remain in trials. Young women may be motivated to enter microbicide trials by stressing the role they will play in discovering better HIV-prevention methods and highlighting the comprehensive preventive exams they will receive.

Fluid balance concepts in medicine:Principles and practice

The regulation of body fluid balance is a key concern in health and disease and comprises three concepts. The first concept pertains to the relationship between total body water(TBW) and total effective solute and is expressed in terms of the tonicity of the body fluids. Disturbances in tonicity are the main factor responsible for changes in cell volume, which can critically affect brain cell function and survival. Solutes distributed almost exclusively in the extracellular compartment(mainly sodium salts) and in the intracellular compartment(mainly potassium salts) contribute to tonicity, while solutes distributed in TBW have no effect on tonicity. The second body fluid balance concept relates to the regulation and measurement of abnormalities of sodium salt balance and extracellular volume. Estimation of extracellular volume is more complex and error prone than measurement of TBW. A key function of extracellular volume, which is defined as the effective arterial blood volume(EABV), is to ensure adequate perfusion of cells and organs. Other factors, including cardiac output, total and regional capacity of both arteries and veins, Starling forces in the capillaries, and gravity also affect the EABV. Collectively, these factors interact closely with extracellular volume and some of them undergo substantial changes in certain acute and chronic severe illnesses. Their changes result not only in extracellular volume expansion, but in the need for a larger extracellular volume compared with that of healthy individuals. Assessing extracellular volume in severe illness is challenging because the estimates of this volume by commonly used methods are prone to large errors in many illnesses. In addition, the optimal extracellular volume may vary from illness to illness, is only partially based on volume measurements by traditional methods, and has not been determined for each illness. Further research is needed to determine optimal extracellular volume levels in several illnesses. For these reasons, extracellular volume in severe illness merits a separate third concept of body fluid balance.

2-甲氧基雌二醇和2-乙氧基雌二醇减少老年肥胖糖尿病ZSF_1大鼠的肾疾病进展

目的检查2-ME和其一个合成类似物2EE(2-ETHOXYESTRADIOL)对高血压、肥胖和肾脏病进展的影响。方法:实验在35周肥胖ZSF_1大鼠中进行。动物分对照组(PEG-400,0.5μl/ h)和治疗组,2ME,2EE(18μg/kg/h),在0周,3,6和9周时分别测定代谢和肾功能,在9周时评价肾血流动力学和排泄功能。结果2ME,2EE对肥胖和高血压无作用,但可降低糖化血红蛋白和糖尿,防止时间依赖的蛋白尿的增加。2ME和2EE显著增加肾血流和肾小球滤过,减少肾血管阻力;免疫标记肾皮质标本,肥胖动物比同窝出生雄瘦动物有显示增加PCNA,NF-KAPPA B和VEGF的表达。2ME,2EE减少PCNA,NF-KAPPA B和VEGF的表达。结论雌二醇的非雌性激素的代谢物2ME和2EE,对代谢病和肥胖相关的肾病有直接的肾脏保护作用。

Mental and physical symptoms associated with lower social support for patients with hepatitis C

AIM: To systematically examine the impact of the hepatitis C virus (HCV) diagnosis on patients' level of social support in a large-scale study. METHODS: Patients evaluated and treated for HCV in a tertiary referral center were enrolled in a cross-sectional study. Demographic data, functional and emotional status as measured by the Hospital Anxiety and Depression Scale (HAD) and the Sickness Impact Profile (SIP), severity of liver disease, mode of acquisition, and physical and psychiatric comorbidities were collected from patients or abstracted from the medical record. All participants completed a semi-structured interview, addressing questions of social support. RESULTS: A total of 342 patients (mean age 45.2 years; 37% women) were enrolled. Ninety-two (27%) patients described lower levels of support by family and friends. Nearly half of the participants (45%) noted the loss of at least one relationship due to the disease. Fears related to transmitting the disease (25%) were common and often associated with ignorance or even discrimination by others (19%). Nearly one fifth of the patients did not share information about their disease with others to avoid being stigmatized. Lower levels of social support were significantly associated with living alone, being unemployed, being excluded from antiviral therapy, having psychiatric comorbidities, contracting HCV through intravenous drug use, having high levels of anxiety and depression as measured by the HAD and negative mood state as measured by the SIP. Patients reporting lower levels of social support also noted more physical symptoms as measured by the SIP. CONCLUSION: Patients with hepatitis C often face significant social problems, ranging from social isolation to familial stress. The most common concerns reflect a limited insight of patients and their relatives and friends about the disease, the risk factors for its spread, and about potential consequences. Our data suggest that educational interventions targeting support persons and the stressors identified in our findings may lessen or alleviate the social strains patients with hepatitis C experience.

Platelet-derived growth factor receptor alpha in glioma:a bad seed

Recent collaborative,large-scale genomic profiling of the most common and aggressive brain tumor glioblastoma multiforme(GBM) has significantly advanced our understanding of this disease.The gene encoding platelet-derived growth factor receptor alpha(PDGFR a) was identified as the third of the top 11 amplified genes in clinical GBM specimens.The important roles of PDGFR a signaling during normal brain development also implicate the possible pathologic consequences of PDGFR a over-activation in glioma.Although the initial clinical trials using PDGFR kinase inhibitors have been predominantly disappointing,diagnostic and treatment modalities involving genomic profiling and personalized medicine are expected to improve the therapy targeting PDGFR a signaling.In this review,we discuss the roles of PDGFR a signaling during development of the normal central nervous system(CNS) and in pathologic conditions such as malignant glioma.We further compare various animal models of PDGF-induced gliomagenesis and their potential as a novel platform of pre-clinical drug testing.We then summarize our recent publication and how these findings will likely impact treatments for gliomas driven by PDGFR a overexpression.A better understanding of PDGFR a signaling in glioma and their microenvironment,through the use of human or mouse models,is necessary to design a more effective therapeutic strategy against gliomas harboring the aberrant PDGFR a signaling.

T1 and ECV Mapping in Myocardial Disease

T1 mapping using cardiovascular magnetic resonance(CMR)introduces novel techniques for myocardial tissue characterization to detect and quantify disease processes occurring at the microscopic level.Even though T1 mapping has limited spatial resolution,cellular and molecular changes occurring within each voxel can affect the aggregate T1 signal rendering them quantifi able.The estimated T1-based parameters quantifi ed on a“map”demonstrate the spatial localization of these changes whereby each pixel expresses the quantitative value of that parameter.This quantifi cation permits detection of diffuse disease even if it is not directly visible.Rather than relying on nonspecifi c functional measures,T1 mapping focuses on intrinsic changes of myocardial composition that advances understanding about specifi c disease pathways.These changes in myocardial tissue composition inform diagnosis and prognosis.T1 mapping encompasses two key parameters:native(i.e.,precontrast)T1 and extracellular volume fraction(ECV)derived from additional postcontrast T1 and blood T1 measurements.These advances introduce new tools to detect focal and diffuse myocardial derangements occurring in cardiac disease that can be otherwise diffi cult to detect.T1 and ECV mapping foster precision medicine and personalized care,promising to improve patient outcomes through targeted therapy.Capitalizing on the opportunities introduced by T1 mapping and ECV requires further investigation.

Hypertonicity:Clinical entities,manifestations and treatment

Hypertonicity causes severe clinical manifestations and is associated with mortality and severe short-term and longterm neurological sequelae. The main clinical syndromes of hypertonicity are hypernatremia and hyperglycemia.Hypernatremia results from relative excess of body sodium over body water. Loss of water in excess of intake,gain of sodium salts in excess of losses or a combination of the two are the main mechanisms of hypernatremia.Hypernatremia can be hypervolemic,euvolemic or hypovolemic. The management of hypernatremia addresses both a quantitative replacement of water and,if present,sodium deficit,and correction of the underlying pathophysiologic process that led to hypernatremia.Hypertonicity in hyperglycemia has two components,solute gain secondary to glucose accumulation in the extracellular compartment and water loss through hyperglycemic osmotic diuresis in excess of the losses of sodium and potassium. Differentiating between these two components of hypertonicity has major therapeutic implications because the first component will be reversed simply by normalization of serum glucose concentration while the second component will require hypotonic fluid replacement. An estimate of the magnitude of the relative water deficit secondary to osmotic diuresis is obtained by the corrected sodium concentration,which represents a calculated value of the serum sodium concentration that would result from reduction of the serum glucose concentration to a normal level.

Immunotherapy for Gastrointestinal Malignancies

Gastrointestinal (GI) malignancies (esophageal, gastric, pancreatic, intra- and extra-biliary ductal, hepatocellular, and colorectal cancers) are an important cause of cancer incidence and mortality in the US and globally. GI cancers account for 15.4% and 23.8% of incident cancers and cancer-related deaths respectively in the US alone. Although earlier diagnosis and treatment advances have improved outcomes for some GI malignancies, the need for improved therapies in all disease phases (adjuvant, neoadjuvant and advanced) is paramount. Utilization of monoclonal antibodies targeting against vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) has shown the success in selected colorectal carcinoma patients. More investigations of immunotherapy are on going in the treatment of GI malignances with different mechanisms and methods. In this article, we review data for established and evolving immunotherapy-related treatment options in GI malignancies.

Sonic hedgehog signaling in kidney fibrosis: a master communicator

The hedgehog signaling cascade is an evolutionarily conserved pathway that regulates multiple aspects of embryonic development and plays a decisive role in tissue homeostasis. As the best studied member of three hedgehog ligands, sonic hedgehog(Shh) is known to be associated with kidney development and tissue repair after various insults. Recent studies uncover an intrinsic link between dysregulated Shh signaling and renal fibrogenesis. In various types of chronic kidney disease(CKD), Shh is upregulated specifically in renal tubular epithelium but targets interstitial fibroblasts, thereby mediating a dynamic epithelialmesenchymal communication(EMC). Tubule-derived Shh acts as a growth factor for interstitial fibroblasts and controls a hierarchy of fibrosis-related genes, which lead to the excessive deposition of extracellular matrix in renal interstitium. In this review, we recapitulate the principle of Shh signaling, its activation and regulation in a variety of kidney diseases. We also discuss the potential mechanisms by which Shh promotes renal fibrosis and assess the efficacy of blocking this signaling in preclinical settings. Continuing these lines of investigations will provide novel opportunities for designing effective therapies to improve CKD prognosis in patients.

Interleukin-6 mediates neutrophil mobilization from bone marrow in pulmonary hypertension

Myeloid cells,such as neutrophils,are produced in the bone marrow in high quantities and are important in the pathogenesis of vascular diseases such as pulmonary hypertension(PH).Although neutrophil recruitment into sites of inflammation has been well studied,the mechanisms of neutrophil egress from the bone marrow are not well understood.Using computational flow cytometry,we observed increased neutrophils in the lungs of patients and mice with PH.Moreover,we found elevated levels of IL-6 in the blood and lungs of patients and mice with PH.We observed that transgenic mice overexpressing Il-6 in the lungs displayed elevated neutrophil egress from the bone marrow and exaggerated neutrophil recruitment to the lungs,resulting in exacerbated pulmonary vascular remodeling,and dysfunctional hemodynamics.Mechanistically,we found that IL-6-induced neutrophil egress from the bone marrow was dependent on interferon regulatory factor 4(IRF-4)-mediated CX3CR1 expression in neutrophils.Consequently,Cx3cr1 genetic deficiency in hematopoietic cells in Il-6-transgenic mice significantly reduced neutrophil egress from bone marrow and decreased neutrophil counts in the lungs,thus ameliorating pulmonary remodeling and hemodynamics.In summary,these findings define a novel mechanism of IL-6-induced neutrophil egress from the bone marrow and reveal a new therapeutic target to curtail neutrophil-mediated inflammation in pulmonary vascular disease.