bluuebunny/crossref_metadata_2025 huggingface dataset:
Inappropriate Prescribing Among Elderly Home Care Patients in Japan
Authors: Hamano Jun , Tokuda Yasuharu
Published: January 2014

Background: Although several previous studies have examined the prevalence and risk factors associated with inappropriate prescribing (IP) among elderly patients, as identified by the Screening Tool of Older Persons’ potentially inappropriate Prescriptions (STOPP) and Screening Tool to Alert doctors to Right Treatment (START) criteria, no research has been conducted among elderly home care patients. Objective: This study aimed to explore the prevalence of IP and its risk factors as identified by the STOPP/START criteria among elderly home care patients in Japan. Study Design: Cross-sectional study. Setting: Home care settings in Japan. Participants: Eighty-nine patients aged 65 years or older who received regular home visiting services from Yamato Clinic between May 2013 and June 2013. Main Outcome Measure: The prevalence of IP and its risk factors as identified by the STOPP/START criteria. Results: Of the study population, 40.4% had at least one potentially inappropriate medication and 60.7% had at least one incidence of underprescribing. Risk factors for potentially inappropriate medications were hypertension, constipation, and polypharmacy, while those for underprescription were osteoporosis and polypharmacy. Conclusion: The prevalence of IP among elderly home care patients is high, with risk factors that include not only polypharmacy but also several specific underlying medical conditions.
Bilateral Anomalous Pulmonary Venous Connection to Bilateral Superior Caval Veins
Authors: Sasikumar Navaneetha , Ramanan Sowmya , Chidambaram Shanthi , Rema Krishna Manohar Soman , Cherian Kootturathu Mammen
Published: January 2014

A four-year-old girl presented with superior vena cava (SVC) type of sinus venosus defect, right upper pulmonary vein draining into right SVC-right atrium junction, left upper lobe pulmonary veins draining into the lower part of persistent left SVC (LSVC), and a patent ductus arteriosus. The anomalous pulmonary venous drainage to LSVC was overlooked in the preoperative evaluation and was found intraoperatively. Warden procedure was performed for right-sided veins. Lower LSVC draining the anomalous pulmonary veins was anastomosed to the left atrial appendage. The short stump of LSVC was diverted to left pulmonary artery. Anomalous pulmonary venous connections to LSVC are rare. Embryology of venous development is analyzed with respect to this rare anomaly, and options for surgical correction are discussed.
Cardiac Rhabdomyoma in a Neonate With Supraventricular Tachycardia
Authors: Sadoh Wilson E. , Obaseki Darlington E. , Amuabunos Emmanuel A. , Eregie Charles O. , Isah Isoken A. , Idemudia Ebenovbe , Vhriterhire Clement O.
Published: January 2014

Cardiac rhabdomyoma (CR) is a rare tumor commonly associated with tuberous sclerosis. They are often detected prenatally or in early infancy. The case of a Nigerian human immunodeficiency virus (HIV)-exposed neonate with CR who presented with supraventricular tachycardia and cardiovascular collapse is presented. The infant was born to a mother on highly active antiretroviral therapy (HAART). The possible role of HIV and HAART in CR etiology and the difficulty in the management of this case are highlighted.
Repeated Aortopulmonary Shunt Thrombosis in a Neonatal Patient With a Low Antithrombin Level
Authors: Niebler Robert A. , Mitchell Michael E. , Scott J. Paul
Published: January 2014

A female infant with unbalanced right ventricular dominant atrioventricular septal defect with double-outlet right ventricle and pulmonary stenosis had recurrent aortopulmonary shunt thrombosis. She was found to have low antithrombin levels and was managed with antithrombin replacement in addition to unfractionated heparin. A subsequent aortopulmonary shunt was successfully placed, and patency was maintained. Her antithrombin levels normalized, and she was continued on low-molecular-weight heparin and aspirin until four months of age when a bidirectional superior cavopulmonary anastomosis was done. A prothrombotic evaluation at the time of the acute thrombosis and repeated at four months of age was negative except for the initially low antithrombin level. A repeat antithrombin level (off supplementation) at the time of the cavopulmonary anastomosis was normal making the diagnosis of congenital antithrombin deficiency unlikely. This case highlights the possibility of neonatal antithrombin deficiency as a cause of aortopulmonary shunt thrombosis and successful management with replacement therapy.
Anomalous Origin of Circumflex Coronary Artery From Right Pulmonary Artery Associated With Coarctation of the Aorta
Authors: Sekelyk Roman , Mykychak Yaroslav , Fedevych Oleh , Yemets Illya
Published: January 2014

Origin of the circumflex coronary artery (Cx) from the pulmonary arteries is an extremely rare anomaly. We describe a two-month-old female patient with anomalous origin of the Cx from the right pulmonary artery associated with coarctation of the aorta. Reimplantation of the anomalous Cx to the aorta and coarctation repair were performed. There were no postoperative complications, and the patient was discharged in satisfactory condition.
Does Single Ventricle Physiology Affect Survival of Children Requiring Extracorporeal Membrane Oxygenation Support Following Cardiac Surgery?
Authors: Alsoufi Bahaaldin , Awan Abid , Manlhiot Cedric , Al-Halees Zohair , Al-Ahmadi Mamdouh , McCrindle Brian W. , Alwadai Abdullah
Published: January 2014

Background:

Improved survival with postoperative extracorporeal membrane oxygenation (ECMO) has expanded its application to children with single ventricle (SV) anomalies. We examine current-era outcomes of postoperative ECMO with special focus on patients with SV.

Methods:

Demographic, anatomic, surgical, and support details of 100 consecutive children requiring postoperative ECMO (2007-2012) were included into multivariable regression models to identify factors affecting survival.

Results:

Median age was 73 days (4 days-16.2 years), 31 patients had SV physiology. The ECMO indication was failure to wean cardiopulmonary bypass (34%) and postoperative low cardiac output (66%) including 37% having extracorporeal cardiopulmonary resuscitation (ECPR). Median ECMO duration was four days (1-21). The ECMO decannulation and survival to hospital discharge were 62% and 37%. In SV group, decannulation and survival rates were 55% and 32%. The SV-ECMO outcomes were best in ECPR subgroup (54%), following shunt (57%) or Norwood (46%) and worst following Glenn, Fontan, or total anomalous pulmonary venous connection repair (0% survival). On multivariable analysis, factors affecting odds of survival were performing angiogram (odds ratio [OR]: 15.28, confidence interval [CI]: 2.34-99.89, P = .004), prolonged ECMO duration (OR: 0.64, CI: 0.47-0.88 per day, P = .005), leaving cannulation snares (OR: 28.41, CI: 2.65-304.70, P = .006), higher HCO3 (OR: 1.19, CI: 1.04-1.36, P = .01), renal failure requiring hemodialysis (OR: 0.21, CI: 0.06-0.76, P = .02), bleeding requiring re-exploration (OR: 0.21, CI: 0.06-0.75, P = .02), ECPR in patients with SV (OR: 11.84, CI: 1.11-126.07, P = .04), delayed lactate normalization (OR: 0.95, CI: 0.90-0.99 per hour, P = .02), and elevated liver enzymes (OR: 0.97, CI: 0.95-1.00 per 10 unit/L, P = .05).

Conclusions:

The ECMO is valuable in patients with SV however results depend on anatomy, procedure, and support indication. Persistent markers of poor perfusion, end-organ injury, and prolonged ECMO duration are associated with mortality. Those factors could be modified by early ECMO application before organ damage, meticulous homeostasis to ensure adequate perfusion, early diagnosis, and reoperation on residual lesions to expedite weaning.
Transthoracic Echocardiographic Predictors of Left Atrial Hypertension in Patients on Venoarterial Extracorporeal Membrane Oxygenation
Authors: Eckhauser Aaron W. , Jones Chris , Witte Madolin K. , Puchalski Michael D.
Published: January 2014

Decompression of the left heart in patients supported with extracorporeal membrane oxygenation (ECMO) is often warranted to protect the myocardium and facilitate recovery. We studied the ability of standard echocardiographic parameters to predict left atrial hypertension by reviewing 3 cardiac patients supported with ECMO who subsequently underwent left atrial decompression. We found that standard echocardiographic parameters poorly predict the need for left atrial decompression on ECMO. Following a more specific diagnostic algorithm to estimate left-sided filling pressure in patients with depressed ejection fraction may significantly improve the ability of echocardiography to accurately predict left atrial hypertension and the need for decompression.
Blowing the Whistle Off the Field of Play
Authors: Richardson Brian K. , McGlynn Joseph
Published: January 2014

Unethical behavior within collegiate sports departments appears to be commonplace. Athletic programs at a number of high-profile universities have been sanctioned by the National Collegiate Athletic Association in the past decade. When athletic departments participate in corrupt behavior, the possibility of whistle-blowing exists. Although organizational scholars have developed theoretical models describing whistle-blowing processes, it is unclear whether these models accurately depict whistle-blower’s experiences in the collegiate sports industry. Thus, the purpose of the present study was to generate a model describing the experiences of whistle-blowers operating in the intercollegiate sports industry. Thirteen whistle-blowers were interviewed for this study; analysis of case studies revealed that whistle-blowing occurred over five stages and was significantly affected by the collegiate athletics context. Of particular consequence was the hypermasculine, highly competitive collegiate sports environment.
The interrelations of radiologic findings and mechanical ventilation in community acquired pneumonia patients admitted to the intensive care unit: a multicentre retrospective study
Authors: Erdem Hakan , Kocak-Tufan Zeliha , Yilmaz Omer , Karakurt Zuhal , Cilli Aykut , Turkan Hulya , Yazicioglu-Mocin Ozlem , Adıguzel Nalan , Gungor Gokay , Taşcı Canturk , Yilmaz Gulden , Oncul Oral , Dogan-Celik Aygul , Erdemli Ozcan , Oztoprak Nefise , Tomak Yakup , Inan Asuman , Tok Demet , Temur Sibel , Oksuz Hafize , Senturk Ozgur , Buyukkocak Unase , Yilmaz-Karadag Fatma , Ozturk-Engin Derya , Ozcengiz Dilek , Karakas Ahmet , Bilgic Hayati , Leblebicioglu Hakan
Published: January 2014

Background

We evaluated patients admitted to the intensive care units with the diagnosis of community acquired pneumonia (CAP) regarding initial radiographic findings.

Methods

A multicenter retrospective study was held. Chest x ray (CXR) and computerized tomography (CT) findings and also their associations with the need of ventilator support were evaluated.

Results

A total of 388 patients were enrolled. Consolidation was the main finding on CXR (89%) and CT (80%) examinations. Of all, 45% had multi-lobar involvement. Bilateral involvement was found in 40% and 44% on CXR and CT respectively. Abscesses and cavitations were rarely found. The highest correlation between CT and CXR findings was observed for interstitial involvement. More than 80% of patients needed ventilator support. Noninvasive mechanical ventilation (NIV) requirement was seen to be more common in those with multi-lobar involvement on CXR as 2.4-fold and consolidation on CT as 47-fold compared with those who do not have these findings. Invasive mechanical ventilation (IMV) need increased 8-fold in patients with multi-lobar involvement on CT.

Conclusion

CXR and CT findings correlate up to a limit in terms of interstitial involvement but not in high percentages in other findings. CAP patients who are admitted to the ICU are severe cases frequently requiring ventilator support. Initial CT and CXR findings may indicate the need for ventilator support, but the assumed ongoing real practice is important and the value of radiologic evaluation beyond clinical findings to predict the mechanical ventilation need is subject for further evaluation with large patient series.
Nickel removal by biosorption onto medlar male flowers coupled with photocatalysis on the spinel ZnMn2O4
Authors: Chergui Ahmed , Madjene Farid , Trari Mohamed , Khouider Ali
Published: January 2014

Ni2+ is a highly toxic above 0.07 mg/L and its removal is of high significance. The biosorption of Ni2+ onto medlar male flowers (MMF) was studied in relation with the physical parameters like pH, contact time, biosorbent dosage, Ni2+ concentration and temperature. The interaction biosorbent-Ni2+ was examined by the FTIR technique. The equilibrium was achieved within 40 min and the data were well fitted by the Langmuir and Redlich-Peterson (R-P) models. The maximum Ni2+ uptake capacity was 17.073 mg/g at 25°C and the Ni2+ removal follows a pseudo-second order kinetic with activation energy of 13.3 kJ/mol. The thermodynamic parameters: ΔS°, ΔH° and ΔG° showed that the biosorption was spontaneous and endothermic. MMF was used as a post treatment technique and the biosorption was coupled with the visible light driven Ni2+ reduction over the spinel ZnMn2O4. The effect of the pH, ZnMn2O4 loading and light intensity on the photoactivity was investigated. 77.5% of Ni2+ was reduced after ~140 min under optimal conditions. The Ni2+ removal reached a rate conversion of 96% of with the coupled system biosorption/photocatalysis is very promising for the water treatment.
Leukemia-Inhibitory Factor—Neuroimmune Modulator of Endocrine Function*
Authors: Auernhammer C. J. , Melmed S.
Published: January 2014

Leukemia-inhibitory factor (LIF) is a pleiotropic cytokine expressed by multiple tissue types. The LIF receptor shares a common gp130 receptor subunit with the IL-6 cytokine superfamily. LIF signaling is mediated mainly by JAK-STAT (janus-kinase-signal transducer and activator of transcription) pathways and is abrogated by the SOCS (suppressor-of cytokine signaling) and PIAS (protein inhibitors of activated STAT) proteins. In addition to classic hematopoietic and neuronal actions, LIF plays a critical role in several endocrine functions including the utero-placental unit, the hypothalamo-pituitary-adrenal axis, bone cell metabolism, energy homeostasis, and hormonally responsive tumors. This paper reviews recent advances in our understanding of molecular mechanisms regulating LIF expression and action and also provides a systemic overview of LIF-mediated endocrine regulation. Local and systemic LIF serve to integrate multiple developmental and functional cell signals, culminating in maintaining appropriate hormonal and metabolic homeostasis. LIF thus functions as a critical molecular interface between the neuroimmune and endocrine systems.
Characterization of ASC-2 as an Anti-Atherogenic Transcriptional Coactivator of Liver X Receptors in Macrophages
Authors: Kim Geun Hyang , Park Keunhee , Yeom Seon-Yong , Lee Kyung Jin , Kim Gukhan , Ko Jesang , Rhee Dong-Kwon , Kim Young Hoon , Lee Hye Kyung , Kim Hae Won , Oh Goo Taeg , Lee Ki-Up , Lee Jae W. , Kim Seung-Whan
Published: January 2014

Activating signal cointegrator-2 (ASC-2) functions as a transcriptional coactivator of many nuclear receptors and also plays important roles in the physiology of the liver and pancreas by interacting with liver X receptors (LXRs), which antagonize the development of atherosclerosis. This study was undertaken to establish the specific function of ASC-2 in macrophages and atherogenesis. Intriguingly, ASC-2 was more highly expressed in macrophages than in the liver and pancreas. To inhibit LXR-specific activity of ASC-2, we used DN2, which contains the C-terminal LXXLL motif of ASC-2 and thereby acts as an LXR-specific, dominant-negative mutant of ASC-2. In DN2-overexpressing transgenic (Tg) macrophages, cellular cholesterol content was higher and cholesterol efflux lower than in control macrophages. DN2 reduced LXR ligand-dependent increases in the levels of ABCA1, ABCG1, and apoE transcripts, as well as the activity of luciferase reporters driven by the LXR response elements (LXREs) of ABCA1, ABCG1, and apoE genes. These inhibitory effects of DN2 were reversed by overexpression of ASC-2. Chromatin immunoprecipitation analysis demonstrated that ASC-2 was recruited to the LXREs of the ABCA1, ABCG1, and apoE genes in a ligand-dependent manner and that DN2 interfered with the recruitment of ASC-2 to these LXREs. Furthermore, low density lipoprotein receptor (LDLR)-null mice receiving bone marrow transplantation from DN2-Tg mice showed accelerated atherogenesis when administered a high-fat diet. Taken together, these results indicate that suppression of the LXR-specific activity of ASC-2 results in both defective cholesterol metabolism in macrophages and accelerated atherogenesis, suggesting that ASC-2 is an anti-atherogenic coactivator of LXRs in macrophages.
Dual Ablation of Grb10 and Grb14 in Mice Reveals Their Combined Role in Regulation of Insulin Signaling and Glucose Homeostasis
Authors: Holt Lowenna J. , Lyons Ruth J. , Ryan Ashleigh S. , Beale Susan M. , Ward Andrew , Cooney Gregory J. , Daly Roger J.
Published: January 2014

Growth factor receptor bound (Grb)10 and Grb14 are closely related adaptor proteins that bind directly to the insulin receptor (IR) and regulate insulin-induced IR tyrosine phosphorylation and signaling to IRS-1 and Akt. Grb10- and Grb14-deficient mice both exhibit improved whole-body glucose homeostasis as a consequence of enhanced insulin signaling and, in the case of the former, altered body composition. However, the combined physiological role of these adaptors has remained undefined. In this study we utilize compound gene knockout mice to demonstrate that although deficiency in one adaptor can enhance insulin-induced IRS-1 phosphorylation and Akt activation, insulin signaling is not increased further upon dual ablation of Grb10 and Grb14. Context-dependent limiting mechanisms appear to include IR hypophosphorylation and decreased IRS-1 expression. In addition, the compound knockouts exhibit an increase in lean mass comparable to Grb10-deficient mice, indicating that this reflects a regulatory function specific to Grb10. However, despite the absence of additive effects on insulin signaling and body composition, the double-knockout mice are protected from the impaired glucose tolerance that results from high-fat feeding, whereas protection is not observed with animals deficient for individual adaptors. These results indicate that, in addition to their described effects on IRS-1/Akt, Grb10 and Grb14 may regulate whole-body glucose homeostasis by additional mechanisms and highlight these adaptors as potential therapeutic targets for amelioration of the insulin resistance associated with type 2 diabetes.
Novel Regulators of Fgf23 Expression and Mineralization in Hyp Bone
Authors: Liu Shiguang , Tang Wen , Fang Jianwen , Ren Jinyu , Li Hua , Xiao Zhousheng , Quarles L. D.
Published: January 2014

We used gene array analysis of cortical bone to identify Phex-dependent gene transcripts associated with abnormal Fgf23 production and mineralization in Hyp mice. We found evidence that elevation of Fgf23 expression in osteocytes is associated with increments in Fgf1, Fgf7, and Egr2 and decrements in Sost, an inhibitor in the Wnt-signaling pathway, were observed in Hyp bone. β-Catenin levels were increased in Hyp cortical bone, and TOPflash luciferase reporter assay showed increased transcriptional activity in Hyp-derived osteoblasts, consistent with Wnt activation. Moreover, activation of Fgf and Wnt-signaling stimulated Fgf23 promoter activity in osteoblasts. We also observed reductions in Bmp1, a metalloproteinase that metabolizes the extracellular matrix protein Dmp1. Alterations were also found in enzymes regulating the posttranslational processing and stability of Fgf23, including decrements in the glycosyltransferase Galnt3 and the proprotein convertase Pcsk5. In addition, we found that the Pcsk5 and the glycosyltransferase Galnt3 were decreased in Hyp bone, suggesting that reduced posttranslational processing of FGF23 may also contribute to increased Fgf23 levels in Hyp mice. With regard to mineralization, we identified additional candidates to explain the intrinsic mineralization defect in Hyp osteoblasts, including increases in the mineralization inhibitors Mgp and Thbs4, as well as increases in local pH-altering factors, carbonic anhydrase 12 (Car12) and 3 (Car3) and the sodium-dependent citrate transporter (Slc13a5). These studies demonstrate the complexity of gene expression alterations in bone that accompanies inactivating Phex mutations and identify novel pathways that may coordinate Fgf23 expression and mineralization of extracellular matrix in Hyp bone.
Genomic Priming of the Antisecretory Response to Estrogen in Rat Distal Colon throughout the Estrous Cycle
Authors: O'Mahony Fiona , Alzamora Rodrigo , Chung Ho-Lam , Thomas Warren , Harvey Brian J.
Published: January 2014

The secretion of Cl− across distal colonic crypt cells provides the driving force for the movement of fluid into the luminal space. 17β-Estradiol (E2) produces a rapid and sustained reduction in secretion in females, which is dependent on the novel protein kinase Cδ (PKCδ) isozyme and PKA isoform I targeting of KCNQ1 channels. This sexual dimorphism in the E2 response is associated with a higher expression level of PKCδ in female compared with the male tissue. The present study revealed the antisecretory response is regulated throughout the female reproductive (estrous) cycle and is primed by genomic regulation of the kinases. E2 (1-10 nm) decreased cAMP-dependent secretion in colonic epithelia during the estrous, metestrous, and diestrous stages. A weak inhibition of secretion was demonstrated in the proestrous stage. The expression levels of PKCδ and PKA fluctuated throughout the estrous cycle and correlated with the potency of the antisecretory effect of E2. The expression of PKCδ and PKA were up-regulated by estrogen at a transcriptional level via a PKCδ-MAPK-cAMP response element-binding protein-regulated pathway indicating a genomic priming of the antisecretory response. PKCδ was activated by the membrane-impermeant E2-BSA, and this response was inhibited by the estrogen receptor antagonist ICI 182,780. The 66-kDa estrogen receptor-α isoform was present at the plasma membrane of female colonic crypt cells with a lower abundance found in male colonic crypts. The study demonstrates estrogen regulation of intestinal secretion both at a rapid and transcriptional level, demonstrating an interdependent relationship between both nongenomic and genomic hormone responses.
Direct Action through the Sertoli Cells Is Essential for Androgen Stimulation of Spermatogenesis
Authors: O'Shaughnessy P. J. , Verhoeven G. , De Gendt K. , Monteiro A. , Abel M. H.
Published: January 2014

Androgens act to stimulate spermatogenesis through androgen receptors (ARs) on the Sertoli cells and peritubular myoid cells. Specific ablation of the AR in either cell type will cause a severe disruption of spermatogenesis. To determine whether androgens can stimulate spermatogenesis through direct action on the peritubular myoid cells alone or whether action on the Sertoli cells is essential, we crossed hypogonadal (hpg) mice that lack gonadotrophins and intratesticular androgen with mice lacking ARs either ubiquitously (ARKO) or specifically on the Sertoli cells (SCARKO). These hpg.ARKO and hpg.SCARKO mice were treated with testosterone (T) or dihydrotestosterone (DHT) for 7 d and testicular morphology and cell numbers assessed. Androgen treatment did not affect Sertoli cell numbers in any animal group. Both T and DHT increased numbers of spermatogonia and spermatocytes in hpg mice, but DHT has no effect on germ cell numbers in hpg.SCARKO and hpg.ARKO mice. T increased germ cell numbers in hpg.SCARKO and hpg.ARKO mice, but this was associated with stimulation of FSH release. Results show that androgen stimulation of spermatogenesis requires direct androgen action on the Sertoli cells.
Decreased Expression of Natriuretic Peptides Associated with Lipid Accumulation in Cardiac Ventricle of Obese Mice
Authors: Bartels Emil Daniel , Nielsen Jan Møller , Bisgaard Line Stattau , Goetze Jens P. , Nielsen Lars Bo
Published: January 2014

Plasma B-type natriuretic peptide (BNP) and proBNP are established markers of cardiac dysfunction. Even though obesity increases the risk of cardiovascular disease, obese individuals have reduced plasma concentrations of natriuretic peptides. The underlying mechanism is not established. We used cultured cardiomyocytes and three different mouse models to examine the impact of obesity and cardiac lipid accumulation on cardiac natriuretic peptide expression. The cardiac ventricular expression of atrial natriuretic peptide (ANP) and BNP mRNA and ANP peptide was decreased 36-72% in obese ob/ob, db/db, and fat-fed C57BL/6 mice as compared with their respective controls. The db/db and ob/ob mice displayed impaired cardiac function, whereas the fat-fed mice had almost normal cardiac function. Moreover, the ventricular expression of hypertrophic genes (α- and β-major histocompatibility complex and α-actin) and natriuretic peptide receptor genes were not consistently altered by obesity across the three mouse models. In contrast, cardiac ventricular triglycerides were similarly increased by 60-115% in all three obese mouse models and incubation with oleic acid caused triglyceride accumulation and an approximately 35% (P < 0.005) depression of ANP mRNA expression in cultured HL-1 atrial myocytes. The data suggest that obesity and altered cardiac lipid metabolism are associated with reduced production of ANP and BNP in the cardiac ventricles in the setting of normal as well as impaired cardiac function.
Estrogen Receptor-β Prevents Cardiac Fibrosis
Authors: Pedram Ali , Razandi Mahnaz , O'Mahony Fiona , Lubahn Dennis , Levin Ellis R.
Published: January 2014

Development of cardiac fibrosis portends the transition and deterioration from hypertrophy to dilation and heart failure. Here we examined how estrogen blocks this important development. Angiotensin II (AngII) and endothelin-1 induce cardiac hypertrophy and fibrosis in humans. and we find that these agents directly stimulate the transition of the cardiac fibroblast to a myofibroblast. AngII and endothelin-1 stimulated TGFβ1 synthesis in the fibroblast, an inducer of fibrosis that signaled via c-jun kinase to Sma- and Mad-related protein 3 phosphorylation and nuclear translocation in myofibroblasts. As a result, mesenchymal proteins fibronectin and vimentin were produced, as were collagens I and III, the major forms found in fibrotic hearts. 17β-Estradiol (E2) or dipropylnitrile, an estrogen receptor (ER)β agonist, comparably blocked all these events, reversed by estrogen receptor (ER)β small interfering RNA. E2 and dipropylnitrile signaling through cAMP and protein kinase A prevented myofibroblast formation and blocked activation of c-jun kinase and important events of fibrosis. In the hearts of ovariectomized female mice, cardiac hypertrophy and fibrosis were induced by AngII infusion and prevented by E2 administration to wild type but not ERβ knockout rodents. Our results establish the cardiac fibroblast as an important target for hypertrophic/fibrosis-inducing peptides the actions of which were mitigated by E2/ERβ acting in these stromal cells.
Identification of Adipocyte Genes Regulated by Caloric Intake
Authors: Franck Niclas , Gummesson Anders , Jernås Margareta , Glad Camilla , Svensson Per-Arne , Guillot Gilles , Rudemo Mats , Nyström Fredrik H , Carlsson Lena M. S. , Olsson Bob
Published: January 2014

Context

Changes in energy intake have marked and rapid effects on metabolic functions, and some of these effects may be due to changes in adipocyte gene expression that precede alterations in body weight.

Objective

The aim of the study was to identify adipocyte genes regulated by changes in caloric intake independent of alterations in body weight.

Research Design and Methods

Obese subjects given a very low-caloric diet followed by gradual reintroduction of ordinary food and healthy subjects subjected to overfeeding were investigated. Adipose tissue biopsies were taken at multiple time-points, and gene expression was measured by DNA microarray. Genes regulated in the obese subjects undergoing caloric restriction followed by refeeding were identified using two-way ANOVA corrected with Bonferroni. From these, genes regulated by caloric restriction and oppositely during the weight-stable refeeding phase were identified in the obese subjects. The genes that were also regulated, in the same direction as the refeeding phase, in the healthy subjects after overfeeding were defined as being regulated by caloric intake. Results were confirmed using real-time PCR or immunoassay.

Results

Using a significance level of P < 0.05 for all comparisons, 52 genes were down-regulated, and 50 were up-regulated by caloric restriction and regulated in the opposite direction by refeeding and overfeeding. Among these were genes involved in lipogenesis (ACLY, ACACA, FASN, SCD), control of protein synthesis (4EBP1, 4EBP2), β-oxidation (CPT1B), and insulin resistance (PEDF, SPARC).

Conclusions

Metabolic genes involved in lipogenesis, protein synthesis, and insulin resistance are central in the transcriptional response of adipocytes to changes in caloric intake.
Altered Autophagy in Human Adipose Tissues in Obesity
Authors: Kovsan Julia , Blüher Matthias , Tarnovscki Tanya , Klöting Nora , Kirshtein Boris , Madar Liron , Shai Iris , Golan Rachel , Harman-Boehm Ilana , Schön Michael R. , Greenberg Andrew S. , Elazar Zvulun , Bashan Nava , Rudich Assaf
Published: January 2014

Context

Autophagy is a housekeeping mechanism, involved in metabolic regulation and stress response, shown recently to regulate lipid droplets biogenesis/breakdown and adipose tissue phenotype.

Objective

We hypothesized that in human obesity autophagy may be altered in adipose tissue in a fat depot and distribution-dependent manner.

Setting and Patients

Paired omental (Om) and sc adipose tissue samples were used from obese and nonobese (n = 65, cohort 1); lean, sc-obese and intraabdominally obese (n = 196, cohort 2); severely obese persons without diabetes or obesity-associated morbidity, matched for being insulin sensitive or resistant (n = 60, cohort 3).

Results

Protein and mRNA levels of the autophagy genes Atg5, LC3A, and LC3B were increased in Om compared with sc, more pronounced among obese persons, particularly if with intraabdominal fat accumulation. Both adipocytes and stromal-vascular cells contribute to the expression of autophagy genes. The increased number of autophagosomes and elevated autophagic flux assessed in fat explants incubated with lysosomal inhibitors were observed in obesity, particularly in Om. The degree of visceral adiposity and adipocyte hypertrophy accounted for approximately 50% of the variance in Atg5 mRNA levels by multivariate regression analysis, whereas age, sex, measures of insulin sensitivity, inflammation, and adipose tissue stress were excluded from the model. Moreover, in cohort 3, the autophagy marker genes were increased in those who were insulin resistant compared with insulin sensitive, particularly in Om.

Conclusions

Autophagy is up-regulated in adipose tissue of obese persons, especially in Om, correlating with the degree of obesity, visceral fat distribution, and adipocyte hypertrophy. This may precede the occurrence of obesity-associated morbidity.
Expanding the Phenotype and Genotype of Female GnRH Deficiency
Authors: Shaw Natalie D. , Seminara Stephanie B. , Welt Corrine K. , Au Margaret G. , Plummer Lacey , Hughes Virginia A. , Dwyer Andrew A. , Martin Kathryn A. , Quinton Richard , Mericq Veronica , Merino Paulina M. , Gusella James F. , Crowley William F. , Pitteloud Nelly , Hall Janet E.
Published: January 2014

Context:

GnRH deficiency is a rare genetic disorder of absent or partial pubertal development. The clinical and genetic characteristics of GnRH-deficient women have not been well-described.

Objective:

To determine the phenotypic and genotypic spectrum of a large series of GnRH-deficient women.

Design, Setting, and Subjects: Retrospective study of 248 females with GnRH deficiency evaluated at an academic medical center between 1980 and 2010.

Main Outcome Measures:

Clinical presentation, baseline endogenous GnRH secretory activity, and DNA sequence variants in 11 genes associated with GnRH deficiency.

Results:

Eighty-eight percent had undergone pubarche, 51% had spontaneous thelarche, and 10% had 1-2 menses. Women with spontaneous thelarche were more likely to demonstrate normal pubarche (P = 0.04). In 27% of women, neuroendocrine studies demonstrated evidence of some endogenous GnRH secretory activity. Thirty-six percent (a large excess relative to controls) harbored a rare sequence variant in a gene associated with GnRH deficiency (87% heterozygous and 13% biallelic), with variants in FGFR1 (15%), GNRHR (6.6%), and PROKR2 (6.6%) being most prevalent. One woman had a biallelic variant in the X-linked gene, KAL1, and nine women had heterozygous variants.

Conclusions:

The clinical presentation of female GnRH deficiency varies from primary amenorrhea and absence of any secondary sexual characteristics to spontaneous breast development and occasional menses. In this cohort, rare sequence variants were present in all of the known genes associated with GnRH deficiency, including the novel identification of GnRH-deficient women with KAL1 variants. The pathogenic mechanism through which KAL1 variants disrupt female reproductive development requires further investigation.
Induction of Carbonic Anhydrase II Expression in Osteoclast Progenitors Requires Physical Contact with Stromal Cells*
Authors: Biskobing Diane M. , Fan Dongjie , Fan Xian , Rubin Janet
Published: January 2014

Carbonic anhydrase II (CA II) expression is vital to normal osteoclast function. We and others have previously reported induction of CA II messenger RNA (mRNA) expression by 1,25(OH)2D3 in myelomonocytic cells and marrow culture. However, since 1,25(OH)2D3 stimulates osteoclast differentiation as well, we wished to separate direct effects of 1,25(OH)2D3 on the CA II gene from the differentiating effects of the hormone. Using primary murine mixed marrow cultures, we measured CA II mRNA expression by RT-PCR. 10 nm 1,25(OH)2D3 dose dependently induced expression of CA II mRNA (4.12 ± 0.68-fold) at day 4 in culture compared with control with an ED50 of 0.25 nm. When nonadherent marrow cells containing osteoclast progenitors were depleted of stromal cells and exposed to 10 nm 1,25(OH)2D3, CA II mRNA expression was decreased by more than 60%. Coculture of progenitors with ST-2 stromal cells for 3 days with 10 nm 1,25(OH)2D3 stimulated CA II expression by 22 ± 3.6-fold. 1,25(OH)2D3 stimulated CA II mRNA expression in progenitors separated from ST-2 cells by transwells was insignificant demonstrating that the two cell types must be in physical contact. PTH also stimulated CA II mRNA expression (4.91 ± 0.01-fold) to a similar degree as seen with 1,25(OH)2D3 treatment. These results demonstrate that induction of CA II in osteoclast progenitors requires their physical communication with stromal cells and is inseparable from the osteoclast differentiation process.
A Role for Phospholipase C Activity in GLUT4-Mediated Glucose Transport*
Authors: Van Epps-Fung Mark , Gupta Kiran , Hardy Robert W. , Wells Alan
Published: January 2014

Overexpression of surrogate receptors [epidermal growth factor (EGF) receptor (EGFR) and platelet-derived growth factor receptor] in adipocytes has demonstrated that multiple signaling pathways may lead to GLUT4-mediated glucose uptake. These implicated pathways function independently of IRS-1 phosphorylation and PI3-kinase activation. In addition, we previously demonstrated that EGFR tyrosyl autophosphorylation is required to stimulate GLUT4-mediated glucose transport in 3T3-L1 adipocytes. This observation suggests that signaling molecules that are dependent on EGFR auto-phosphorylation, such as phospholipase C (PLC), may lie in the signaling pathway to glucose transport. As PLC has been implicated in glucose transport by several clinical and basic mechanistic studies, we investigated whether EGFR signaling may promote glucose transport via modulation of PLC activity. Activation of EGFR overexpressing 3T3-L1 adipocytes leads to a 3.4 ± 1.2-fold stimulation of PLC activity over basal levels vs. only 1.06 ± 0.01-fold stimulation by insulin. Pharmacological inhibition of PLC by 50 μm U73122 reduced phosphoinositide accumulation by 79.2 ± 16.9% and resulted in a concomitant 56.0 ± 12.7% decrease in EGF-induced glucose transport. This inhibition of glucose transport by U73122 was specific, because the inactive congener, U73343, failed to block EGF-induced glucose transport. Despite the low levels of insulin-induced PLC activity, insulin-stimulated glucose transport activity was similarly inhibited by U73122 (55.9 ± 13.1% inhibition). Inhibition of PLC activation did not impair either EGF- or insulin-induced activation of glycogen synthase or incorporation of glucose into lipid, supporting the hypothesis that both EGF- and insulin-induced glucose disposal can be independent of GLUT4-mediated glucose transport. The diminution of glucose transport secondary to inhibition of PLC activity was reflected by a decrease in GLUT4 translocation to the plasma membrane upon either EGF or insulin stimulation. These results are consistent with either a permissive or an active role for PLC activity in the translocation of GLUT4 to the plasma membrane.
Insulin-Like Growth Factor I (IGF-I) Receptor Overexpression Abolishes the IGF Requirement for Differentiation and Induces a Ligand-Dependent Transformed Phenotype in C2 Inducible Myoblasts*
Authors: Navarro Magali , Barenton Bruno , Garandel Veronique , Schnekenburger Juergen , Bernardi Henri
Published: January 2014

Insulin-like growth factors (IGFs) stimulate both proliferation and differentiation of myogenic cell lines, and these actions are mostly mediated through the type I IGF receptor (type I IGF-R). To further investigate the role of this receptor in phenotypic characteristics of C2 murine myoblasts, we overexpressed the human type I IGF-R in the inducible clone of C2 cells, which requires IGFs in the differentiation medium to undergo terminal differentiation.

Inducible myoblasts were transfected with either the eukaryotic expression vector pNTK or pNTK containing the human type I IGF-R complementary DNA, and we isolated two clones named Ind-Neo and Ind-R, respectively. Binding and autophosphorylation experiments indicate that Ind-R cells express about 10 times as much type I IGF-R compared with Ind-Neo control cells and that the transfected type I IGF-R is functional in Ind-R cells.

We show that overexpression of the human type I IGF-R makes inducible myoblasts able to differentiate spontaneously, as assessed by expression of the myogenic transcription factors MyoD and myogenin, detection of the muscle-specific protein troponin T, and myotube formation. Moreover, when exposed to IGF-I, Ind-R cells lose contact inhibition, grow in the presence of a low level of growth factors and form colonies in soft agar, which is characteristic of a ligand-dependent transformed phenotype.

It emerges from this study that 1) the type I IGF-R is strongly involved in the phenotypic differences between inducible and permissive cells with respect to the differentiation program; and 2) overexpression causes this receptor to act as a ligand-dependent transforming protein in muscle cells. We suggest that type I IGF-R abundance and level of activation may determine the efficiency of the autocrine mode of action of IGFs and discriminate their biological functions.
The Type 2 and Type 3 Iodothyronine Deiodinases Play Important Roles in Coordinating Development in Rana catesbeiana Tadpoles*
Authors: Becker Kathryn B. , Stephens Kristen C. , Davey Jennifer C. , Schneider Mark J. , Galton Valerie Anne
Published: January 2014

In developing Rana catesbeiana tadpoles, the timing of the thyroid hormone (TH)-dependent metamorphic responses varies markedly among tissues. Yet at any one time these tissues are exposed to the same plasma concentration of TH, suggesting that TH action is regulated in part at the level of the peripheral tissues. A major factor in TH action is the intracellular level of the active TH, T3. This level is dependent not only on the plasma concentration of TH (mostly T4) but also on the intracellular activities of the type 2 5′-deiodinase (D2) and the type 3 5-deiodinase (D3), which are responsible, respectively, for generating and degrading T3. (D1 is not present in this species.) To determine whether differential expression of D2 and D3 among tissues could be a significant factor in the coordination of metamorphic events, the ontogenic profiles of the two enzyme activities and corresponding messenger RNA levels in most tissues of R. catesbeiana tadpoles have been documented. The profiles of D2 expression in tail, hindlimb, forelimb, intestine, skin, and eye differed markedly at both activity and messenger RNA levels, but it was notable that expression was invariably highest in a given tissue at the time of its major metamorphic change. D2 expression was very low in brain and heart and did not vary during development. D2 was not expressed in liver, kidney, or red blood cells. With the exception of red blood cells, D3 expression was detected in all tissues studied. Furthermore, it was evident that in tissues that expressed both deiodinase genes, the two expression profiles were comparable, indicating a potential for tight control of intracellular T3 levels.

Direct evidence of the importance of the intracellular conversion of T4 to T3 for TH-dependent metamorphic events was obtained in tadpoles in which endogenous TH synthesis was blocked with methimazole, and the activities of D2 and D3 were inhibited by iopanoic acid. This treatment inhibited metamorphosis. The inhibition could be overcome by the concomitant administration of replacement levels of T3, but not T4.

These results strongly support the view that coordinated development in amphibia depends in part on the tissue-specific expression patterns of the D2 and D3 genes, which ensure that the requisite level of intracellular T3 is attained in a given tissue, regardless of the current level of circulating TH, at the appropriate stage of metamorphosis.
Immunization of Male Bonnet Monkeys (M. radiata) with a Recombinant FSH Receptor Preparation Affects Testicular Function and Fertility
Authors: Moudgal N. R. , Sairam M. R. , Krishnamurthy H. N. , Sridhar Surekha , Krishnamurthy H. , Khan H.
Published: January 2014

Immunization of proven fertile adult male monkeys (n = 3) with a recombinant FSH receptor protein preparation (oFSHR-P) (representing amino acids 1–134 of the extracellular domain of the receptor Mr∼ 15KDa) resulted in production of receptor blocking antibodies. The ability of the antibody to bind a particulate FSH receptor preparation and receptors in intact granulosa cells was markedly (by 30–80%) inhibited by FSH. Serum T levels and LH receptor function following immunization remained unchanged. The immunized monkeys showed a 50% reduction (p< 0.001) in transformation of spermatogonia (2C) to primary spermatocytes (4C) as determined by flow cytometry and the 4C:2C ratio showed a correlative change (R 0.81, p< 0.0007) with reduction in fertility index (sperm counts X motility score). Breeding studies indicated that monkeys became infertile between 242–368 days of immunization when the fertility index was in the range of 123 ± 76 to 354 ± 42 (compared to a value of 1602 ± 384 on day 0). As the effects observed are near identical to that seen following immunization with FSH it is suggestive that oFSHR-P can substitute for FSH in the development of a contraceptive vaccine.
Ligand-Independent GLUT4 Translocation Induced by Guanosine 5′-O-(3-Thiotriphosphate) Involves Tyrosine Phosphorylation*
Authors: Haruta Tetsuro , Morris Aaron J. , Vollenweider Peter , Nelson James G. , Rose David W. , Mueckler Michael , Olefsky Jerrold M.
Published: January 2014

To delineate the signaling pathway leading to glucose transport protein (GLUT4) translocation, we examined the effect of microinjection of the nonhydrolyzable GTP analog, guanosine 5′-O-(3-thiotriphosphate) (GTPγS), into 3T3-L1 adipocytes. Thirty minutes after the injection of 5 mm GTPγS, 40% of injected cells displayed surface GLUT4 staining indicative of GLUT4 translocation compared with 55% for insulin-treated cells and 10% in control IgG-injected cells. Treatment of the cells with the phosphatidylinositol 3-kinase inhibitor wortmannin or coinjection of GST-p85 SH2 fusion protein had no effect on GTPγS-mediated GLUT4 translocation. On the other hand, coinjection of antiphosphotyrosine antibodies (PY20) blocked GTPγS-induced GLUT4 translocation by 65%. Furthermore, microinjection of GTPγS led to the appearance of tyrosine-phosphorylated proteins around the periphery of the plasma membrane, as observed by immunostaining with PY20. Treatment of the cells with insulin caused a similar phosphotyrosine-staining pattern. Electroporation of GTPγS stimulated 2-deoxy-d-glucose transport to 70% of the extent of insulin stimulation. In addition, immunoblotting with phosphotyrosine antibodies after electroporation of GTPγS revealed increased tyrosine phosphorylation of several proteins, including 70- to 80-kDa and 120- to 130-kDa species. These results suggest that GTPγS acts upon a signaling pathway either downstream of or parallel to activation of phosphatidylinositol 3-kinase and that this pathway involves tyrosine-phosphorylated protein(s).
A Minimum of Fuel Is Necessary for Tolbutamide to Mimic the Effects of Glucose on Electrical Activity in Pancreatic β-Cells*
Authors: Henquin Jean-Claude
Published: January 2014

Glucose stimulation of pancreatic β-cells triggers electrical activity (slow waves of membrane potential with superimposed spikes) that is best monitored with intracellular microelectrodes. Closure of ATP-sensitive K+ channels underlies the depolarization to the threshold potential and participates in the increase in electrical activity produced by suprathreshold (>7 mm) concentrations of glucose, but it is still unclear whether this is the sole mechanism of control. This was investigated by testing whether blockade of ATP-sensitive K+ channels by low concentrations of tolbutamide is able to mimic the effects of glucose on mouse β-cell electrical activity even in the absence of the sugar. The response to tolbutamide was influenced by the duration of the perifusion with the low glucose medium. Tolbutamide (25 μm) caused a rapid and sustained depolarization with continuous activity after 6 min of perifusion of the islet with 3 mm glucose, and a progressive depolarization with slow waves of the membrane potential after 20 min. In the absence of glucose, the β-cell response to tolbutamide was a transient phase of depolarization with rare slow waves (6 min) or a silent, small, but sustained, depolarization (20 min). Readministration of 3 mm glucose was sufficient to restore slow waves, whereas an increase in the glucose concentration to 5 and 7 mm was followed by a lengthening of the slow waves and a shortening of the intervals. In contrast, induction of slow waves by tolbutamide proved very difficult in the absence of glucose, because the β-cell membrane tended to depolarize from a silent level to the plateau level, at which electrical activity is continuous. Azide, a mitochondrial poison, abrogated the electrical activity induced by tolbutamide in the absence of glucose, which demonstrates the influence of the metabolism of endogenous fuels on the response to the sulfonylurea. The partial repolarization that azide also produced was reversed by increasing the concentration of tolbutamide, but reappearance of the spikes required the addition of glucose. It is concluded that inhibition of ATP-sensitive K+ channels is not the only mechanism by which glucose controls electrical activity inβ -cells.