bluuebunny/crossref_metadata_2025 huggingface dataset:
Establishment of self-sequential longitudinal reference intervals of maternal thyroid function during pregnancy
Authors: Yu Bin , Wang Qiu-Wei , Huang Rui-Ping , Cao Fang , Zhu Zi-Qiang , Sun Da-Cheng , Zhou Hong , Zhang Yi-Ming
Published: April 2011

The objective of this study is to establish self-sequential longitudinal reference intervals of thyroid function in normal pregnant women. According to the selection criteria, 301 cases were taken as the normal pregnant population to establish a normal reference range. Meanwhile, 150 healthy women were selected as the normal non-pregnant control group. To establish their own self-sequential longitudinal reference intervals, we collected samples five times in every case throughout the gestation (including first trimester, second trimester, third trimester, prenatal and postpartum), and detected the levels of thyroid stimulating hormone (TSH), free thyroxine (FT4), thyroid peroxidase antibodies (TPO-Ab), and then established the self-sequential longitudinal reference intervals. The levels of TSH, FT4 and TPO-Ab were quantified by electrochemistry immunoassay (ECL) and statistically analyzed using SPSS 13.0 software. Serum TSH of normal pregnant women was at a low level in the first trimester ( P < 0.05) and began to rise continuously. Not until prenatal phase was it restored to the non-pregnant state ( P > 0.05). During pregnancy, serum FT4 of normal pregnant women were consistently lower than non-pregnant levels ( P < 0.05) and kept at low levels. Serum TPO-Ab increased significantly in the third trimester and prenatal phase ( P < 0.05). Of normal pregnant women, 6.5% were TPO-Ab positive.

In conclusion, the reference intervals in our case will reflect the changes of thyroid function in pregnant women more realistically, resulting in a more accurate value for clinical diagnosis and therapy.
Regulation of cell proliferation by the opioid growth factor receptor is dependent on karyopherinβand Ran for nucleocytoplasmic trafficking
Authors: Cheng Fan , McLaughlin Patricia J , Zagon Ian S
Published: April 2011

The opioid growth factor (OGF; [Met5]-enkephalin) and the OGF receptor (OGFr) form an endogenous and tonically active growth-regulating system that modulates cell proliferation by upregulating the cyclin-dependent kinase inhibitory pathway. Previous reports have documented that nucleocytoplasmic trafficking of the OGF–OGFr axis is dependent on nuclear localization signals. This study determined the specific transport factors required for the import of the OGF–OGFr complex from the cytoplasm to the nucleus using a probe of full-length OGFr fused to enhanced green fluorescent protein (eGFP) and knockdown of karyopherin α1, α2, α3, α4 or α6, karyopherin β1 or Ran with small interfering RNA (siRNA). A human squamous cell carcinoma of the head and neck cell line (squamous cell carcinoma-1, SCC-1) that was downregulated for karyopherin β1 or Ran did not have transport of OGFr-eGFP into the nucleus. Moreover, there was an increase of 44% in bromodeoxyuridine (BrdU)-labeled cells in cultures of SCC-1 that were transfected with siRNAs to karyopherin β1 or Ran compared with cells transfected with scrambled siRNA. No alteration in distribution of OGFr-eGFP or BrdU labeling indexes was recorded in cultures treated with siRNAs to karyopherin α1, α2, α3, α4 or α6. These results indicate that the regulation of cell proliferation by the OGF–OGFr axis is dependent on nucleocytoplasmic transport by karyopherin β1 as well as the gradient of RanGTP/RanGDP across the nuclear envelope, but is not reliant on adaptor molecules related to karyopherin α. Thus, the passage of the OGF–OGFr complex has controlled entry from the cytoplasm to the nucleus, and the timely and faithful translocation of this cargo across the nuclear envelope is critical to cell proliferation. These hierarchical levels of nuclear import provide multiple pathways for the subtle regulation of OGF–OGFr as it relates to the control of cell proliferative events.
Overexpression of histone variant H2A.1 and cellular transformation are related in N-nitrosodiethylamine-induced sequential hepatocarcinogenesis
Authors: Khare Satyajeet P , Sharma Ajitkumar , Deodhar Kedar K , Gupta Sanjay
Published: April 2011

Histones through a complex repertoire of non-allelic variants and their post-translational modifications regulate gene expression. Though alterations in histone-modifying enzymes and post-translational modifications of histones have been studied in cancer, expression of histone variants has not been clearly associated with dedifferentiation and malignant transformation of hepatocyte in vivo. In the present work, the pattern of variants of histones was investigated during N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis. Our studies show for the first time in vivo overexpression of a major histone H2A variant H2A.1 and a decrease in H2A.2 at protein and mRNA levels by sodium dodecyl sulfate-Acetic acid-Urea-Triton (SDS-AUT) two-dimensional gel electrophoresis followed by matrix-assisted-laser desorption/ionization time-of-flight (TOF)/TOF mass spectrometry and reverse transcriptase-polymerase chain reaction analysis during sequential development of hepatocellular carcinoma (HCC). H2A.1 and H2A.2 are highly homologous, replication-dependent, non-allelic variants of histone H2A differing at only three amino acid positions. Our results of increase in proliferating cell nuclear antigen expression indicate that with increase in replicating population of transformed cells in HCC, H2A.1 expression increases, suggesting association of H2A.1 overexpression with hyper-proliferation of hepatocytes during cellular dedifferentiation and progressive transformation of normal liver to preneoplastic and neoplastic stages of HCC.
Carbon monoxide inhalation decreased lung injury via anti-inflammatory and anti-apoptotic effects in brain death rats
Authors: Zhou Huacheng , Liu Jinfeng , Pan Peng , Jin Di , Ding Wengang , Li Wenzhi
Published: April 2011

Brain death (BD) induces acute lung injury and makes donor lungs unfit for transplantation. Carbon monoxide (CO) inhalation at 50–500 ppm exerts anti-inflammatory and anti-apoptosis effects in several lung injury models. We examined whether CO inhalation would show favorable effects on lung injury in BD rats. BD rats inhaled 250 ppm CO for two hours. Inhalation decreased the severity of lung injury, as checked by histological examination. CO treatment reversed aggravation in PaO2/FiO2, base excess and pH of BD rats. CO inhalation downregulated the pro-inflammatory cytokines (tumor necrosis factor- α, interleukin-6), and inhibited activity of myeloperoxidase in lung tissue. Inhalation significantly decreased cell apoptosis of lungs, and inhibited mRNA expression of intercellular adhesion molecule-1 and caspase-3 in the lungs. Further, the inhalation activated phosphorylation of p38 expression and inhibited phosphorylation of extracellular signal-regulated kinase expression in the lungs. In conclusion, CO exerts potent protective effects on lungs from BD rats, exhibiting anti-inflammatory and anti-apoptosis functions by modulating the mitogen-activated protein kinase signal transduction.
Relationship between brachial and femoral artery endothelial vasomotor function/phenotype in pigs
Authors: Padilla Jaume , Simmons Grant H , Newcomer Sean C , Laughlin M Harold
Published: April 2011

In humans, the measurement of brachial artery endothelial vasomotor function is used as a surrogate index of systemic endothelial health; however, the applicability of brachial artery findings to other vasculatures needs to be examined. The purpose of the present investigation was to test the following hypotheses: (1) brachial and femoral artery endothelium-dependent/independent relaxation is correlated; (2) endothelial expression of pro-/antiatherogenic proteins is correlated between brachial and femoral arteries; and (3) within vessel, there is a positive correlation between expression of antiatherogenic proteins and endothelium-dependent/independent relaxation, and an inverse correlation between expression of proatherogenic proteins and relaxation. In vitro endothelium-dependent (bradykinin [BK] and acetylcholine [Ach]) and -independent (sodium nitroprusside [SNP]) relaxation were evaluated in harvested brachial and femoral arteries of 96 Yucatan miniature swine. In a subset of pigs ( n = 32), expression of 18 pro-/antiatherogenic proteins was measured from brachial and femoral artery endothelial cell scrapes using immunoblot analysis. Vascular sensitivity (half-maximal effective dose) to BK, Ach and SNP was highly correlated between brachial and femoral arteries ( P < 0.01). A significant correlation was found between brachial and femoral arteries for content of six of the 18 measured proteins ( P < 0.01). Furthermore, expression of two proteins (eNOS and COX-1) was correlated with vasorelaxation function in the brachial artery ( P < 0.01). We provide the first evidence of a relationship between brachial and femoral artery endothelium-dependent relaxation. Our data also suggest that, in general terms, endothelial expression of several established pro-/antiatherogenic proteins is not robustly associated between brachial and femoral arteries, and does not link strongly to vasorelaxation function.
Distinct effects of intravascular and extravascular angiotensin II on cerebrovascular circulation of newborn pigs
Authors: Knecht Kenneth R , Leffler Charles W
Published: April 2011

Angiotensin II (AngII) is important in regulation of vascular resistance and control of blood flow among organs and tissues. The effect of AngII on the cerebral microvasculature may be mediated or altered by endothelial-derived signals. The aim of this study was to test the hypothesis that blood AngII dilates neonatal pial arterioles via an endothelial-dependent mechanism but brain AngII can constrict pial arterioles by activating smooth muscle AT1 receptors. Studies used anesthetized newborn pigs with surgically implanted closed cranial windows. AngII was given either by infusion into the carotid artery ipsilateral to the cranial window or topically. Intracarotid infusion of AngII dilated pial arterioles. The dilation was blocked by systemic administration of the AT1-receptor antagonist, losartan, but unaffected by topical losartan. Topical AngII also caused dilation, but this dilation was converted to constriction by topical losartan. In piglets pretreated with the angiotensin-converting enzyme (ACE) inhibitor, enalapril, topical AngII constricted, rather than dilated, pial arterioles. In enalapril-treated piglets, light/dye endothelial injury blocked dilation to intracarotid AngII but did not affect constriction to topical AngII. Either indomethacin or l-nitroarginine methyl ester blocked the dilation to intraluminal AngII, but neither affected constriction to topical AngII. Chromium mesoporphyrin, that inhibits heme oxygenase, did not affect responses to either topical or intravascular AngII. These data are consistent with the hypotheses that: (a) circulating AngII dilates pial arterioles via endothelial AT1 receptor-derived relaxing factors, notably prostanoids and nitric oxide; (b) direct AT1 receptor activation on the brain side of the blood–brain barrier by AngII causes AT1 receptor-mediated constriction that can mask underlying AT1 receptor-independent dilation when ACE is inhibited. Clinical manipulation of the renin–angiotensin system will have disparate actions on cerebral circulation depending on the functional integrity of the intima and ACE.
Angiogenic effect of laminin involves modulation of cyclooxygenase-2 and prostaglandin levels
Authors: Kumar V B Sameer , Viji Raveendran I , Kiran Manikantan Syamala , Sudhakaran Perumana R
Published: April 2011

The molecular mechanism of the angiogenic effect of laminin (Ln) was studied using human umbilical vein endothelial cells (HUVECs) maintained in culture on Ln-1 substratum. High-pressure liquid chromatography analysis showed that in cells maintained on Ln, the levels of proangiogenic prostaglandin E2(PGE2) increased and that of antiangiogenic PGD2decreased. The angiogenic effect of PGE2and PGD2was confirmed by assessing the expression of CD31 and E-selectin in HUVECs. Immunoblot analysis, reverse transcription-polymerase chain reaction and cyclooxygenase (COX) assay showed increase in the expression and activity of COX-2 in cells maintained on Ln. Use of pharmacological inhibitors suggested that the modulation in the expression of COX-2 and thereby the levels of PGE2and PGD2in endothelial cells by Ln is mediated through the α6β4integrin-p38MAPK (mitogen-activated protein kinase)–NF-κB signaling pathway.
Mitotic recombination: a genotoxic effect of the antidepressant citalopram in Aspergillus nidulans
Authors: Franco Claudinéia C S , Castro-Prado Juliana , Rosada Lúcia J , Sant'Anna Juliane R , Castro-Prado Marialba A A
Published: April 2011

This report evaluates the potential of the antidepressant drug citalopram to induce homozygotization of genes previously present in a heterozygous condition, by homologous recombination. In order to address this question, a heterozygous diploid strain of the filamentous fungus Aspergillus nidulans and the homozygotization assay were utilized. Non-cytotoxic concentrations of citalopram (50, 75 and 100 μmol/L) showed a strong recombinogenic effect in A. nidulans, inducing homozygosis of the diploid strain's nutritional markers. The genetic markers exhibited homozygotization index (HI) rates higher than 2.0 and significantly different from HI control ones. Since citalopram has been previously characterized as a DNA synthesis inhibitor, the recombinogenic potential of this antidepressant in A. nidulans may be associated with the recombinational repair of citalopram-induced DNA strand breaks.
Extracellular signal-regulated kinases 1/2 suppression aggravates transforming growth factor-beta1 hepatotoxicity: a potential mechanism for liver injury in methionine-choline deficient-diet-fed mice
Authors: Wang Zhigang , Yao Tong , Song Zhenyuan
Published: April 2011

Hepatocyte cell death is a characteristic indication in the development of non-alcoholic steatohepatitis (NASH); however, the underlying mechanism is still unclear. In this study, we examined the potential mechanism(s) involved in the development of liver injury using a methionine-choline deficient (MCD) diet feeding NASH model. Male C57BL6/J mice were fed MCD and methionine-choline sufficient (MCS) diet for two weeks before being killed. Our results showed that MCD diet feeding resulted in fatty liver and liver injury, evidenced by increased hepatic triglyceride (TG), plasma alanine aminotransferases and hepatic thiobarbituric acid reactive substances levels in MCD-fed mice. Furthermore, we found that MCD diet feeding caused remarkable suppression of hepatic extracellular signal-regulated kinases (ERK) 1/2 activation and increased transforming growth factor (TGF)-beta1 levels in plasma and the liver tissue. In vitro investigations showed that intracellular MEK/ERK1/2 activation status played a critical role in the determination of sensitivity of hepatocytes to TGF-beta1-induced cell death. HepG2 cells, otherwise resistant to TGF-beta1 killing due to high level of ERK1/2 activation, was sensitized by U0126, a specific MEK/ERK1/2 inhibitor, to TGF-beta1 cytotoxicity. H4IIEC3 cells, which have lower level of constitutive ERK1/2 activity, are sensitive to TGF-beta1-induced cell death. Lastly, we demonstrated that administration of epidermal growth factor, a strong ERK1/2 activator, to MCD-fed mice attenuated liver injury without affecting hepatic TG accumulation. Our findings demonstrated that hepatic ERK1/2 inactivation aggravates TGF-beta1-induced hepatotoxicity, which may contribute, at least in part, to the initiation of liver injury in NASH.
Sitagliptin lowers glucagon and improves glucose tolerance in prediabetic obese SHROB rats
Authors: Chen Brian , Moore Alex , Escobedo Liza V S , Koletsky Matthew S , Hou Diana , Koletsky Richard J , Ernsberger Paul
Published: April 2011

The SHROB (spontaneously hypertensive rat – obese strain) is a model of prediabetes and metabolic syndrome with insulin resistance, glucose intolerance and hypertension. Inhibitors of dipeptidyl dipeptidase IV (DPP-IV) are effective hypoglycemic agents in type 2 diabetes through potentiation of incretin hormones that act in the pancreas to increase insulin and decrease glucagon release. We sought to determine whether the DPP-IV inhibitor sitagliptin might be effective in prediabetes relative to standard therapy with the sulfonylurea glyburide, by using the SHROB model. SHROB show normal fasting glucose but are insulin resistant and hyperglucagonemic. SHROB were treated for six weeks with vehicle, sitagliptin (30 mg/kg/d) or glyburide (1 mg/kg/d) and compared with untreated lean spontaneously hypertensive rats. Body weight, food intake and fasting glucose were all unchanged in all three SHROB groups, but glucagon was reduced by 33% by sitagliptin while remaining unchanged following glyburide or vehicle. In oral glucose (6 g/kg) tolerance testing, both sitagliptin and glyburide lowered plasma glucose. Both sitagliptin and glyburide shifted peak insulin secretion earlier (30 min for glyburide and 60 min for sitagliptin but 240 min for vehicle). Only sitagliptin significantly enhanced insulin secretion. Sitagliptin is effective in normalizing excess glucagon levels and delaying exaggerated insulin secretion in response to a glucose challenge in a prediabetic model.
Fasting-induced reductions in cardiovascular and metabolic variables occur sooner in obese versus lean mice
Authors: Tanner Jason M , Kearns Devin T , Kim Bum Jun , Sloan Crystal , Jia Zhanjun , Yang Tianxin , Abel E Dale , Symons J David
Published: April 2011

It is not uncommon for laboratory animals to be fasted prior to experimentation. Fasting evokes marked reductions in heart rate (HR), blood pressure (BP), heat production and oxygen consumption (VO2) in rodents. Mice with diet-induced obesity exhibit elevated HR and BP, and lower VO2and heat production in the fed condition versus their lean counterparts. It is unknown whether body composition alters the tempo of response to fasting. We tested the hypothesis that cardiovascular and metabolic responses to fasting are delayed in obese versus lean male C57BL/6J mice. In the fed condition, mice that consumed high-fat (HF, 45% fat) chow for 98 ± 5 days had elevated ( P < 0.05) body fat percentage (DEXA), serum leptin (ELISA), HR and BP (72-h biotelemetry), and lower ( P < 0.05) heat production and VO2(72-h metabolic chamber) versus animals that consumed standard chow (CON, 10% fat; n = 16 per group). HR, BP, VO2, heat production and serum leptin decreased (all P < 0.05) in response to a 16-h fast (16:00–08:00 h) in both groups. Although the overall fold changes in cardiovascular and metabolic parameters were similar in magnitude among animals, fasting-induced reductions in cardiovascular and metabolic variables occurred ∼4 and ∼7 h earlier ( P < 0.05), respectively, in HF versus CON mice. These findings indicate that while metabolic and cardiovascular stress evoked by a 16-h fast at 22°C is not different between HF and CON mice, fasting-induced responses occur sooner in obese animals.
Psychological stress during exercise: immunoendocrine and oxidative responses
Authors: Huang Chun-Jung , Webb Heather E , Evans Ronald K , McCleod Kelly A , Tangsilsat Supatchara E , Kamimori Gary H , Acevedo Edmund O
Published: April 2011

The purpose of this study was to examine the changes in catecholamines (epinephrine [EPI] and norepinephrine [NE]), interleukin-2 (IL-2) and a biomarker of oxidative stress (8-isoprostane) in healthy individuals who were exposed to a dual challenge (physical and psychological stress). Furthermore, this study also examined the possible relationships between catecholamines (NE and EPI) and 8-isoprostane and between IL-2 and 8-isoprostane following a combined physical and psychological challenge. Seven healthy male subjects completed two experimental conditions. The exercise-alone condition (EAC) consisted of cycling at 60% VO2max for 37 min, while the dual-stress condition (DSC) included 20 min of a mental challenge while cycling. DSC showed greater EPI and 8-isoprostane levels (significant condition by time interaction). NE and IL-2 revealed significant change across time in both conditions. In addition, following dual stress, EPI area-under-the-curve (AUC) demonstrated a positive correlation with NE AUC and IL-2 AUC. NE AUC was positively correlated with IL-2 AUC and peak 8-isoprostane, and peak IL-2 was positively correlated with peak 8-isoprostane in response to a dual stress. The potential explanation for elevated oxidative stress during dual stress may be through the effects of the release of catecholamines and IL-2. These findings may further provide the potential explanation that dual stress alters physiological homeostasis in many occupations including firefighting, military operations and law enforcement. A greater understanding of these responses to stress can assist in finding strategies (e.g. exercise training) to overcome the inherent psychobiological challenges associated with physically and mentally demanding professions.
A ketogenic diet impairs energy and glucose homeostasis by the attenuation of hypothalamic leptin signaling and hepatic insulin signaling in a rat model of non-obese type 2 diabetes
Authors: Park Sunmin , Kim Da Sol , Kang Sunna , Daily James W
Published: April 2011

Ketogenic diets (KTD) are reported to have beneficial effects on the regulation of energy and glucose homeostasis, but remain controversial. We investigated the effects of KTD and ketones on insulin resistance and secretion in non-obese type 2 diabetic rats and their mechanism. KTD (82% energy as fat), intraperitoneal injection of β-hydroxybutyrate (IHB; 150 mg/kg bw/12 h) with a control diet (COD; 20% energy as fat) or saline injection with COD was given to 90% pancreatectomized (Px) diabetic rats for five weeks. KTD increased epididymal fat pads and serum leptin levels without increasing energy intake, but IHB decreased them. KTD, but not IHB, attenuated hypothalamic signal transducer and activator of transcription 3 and 5′-adenosine monophosphate-activated protein kinase (AMPK) phosphorylation in KTD. Serum glucagon levels were markedly higher in the KTD group than in other groups. During an oral glucose tolerance test, serum glucose levels slowly increased until 80 min in the KTD group and then decreased very slowly. Insulin secretion capacity during a hyperglycemic clamp was significantly lower in the IHB group than in other groups. However, a euglycemic hyperinsulinemic clamp revealed that KTD decreased glucose infusion rates and increased hepatic glucose output in hyperinsulinemic states while IHB had opposite effects to KTD. The increased hepatic glucose output in KTD was associated with increased hepatic phosphoenolpyruvate carboxykinase expression through attenuated tyrosine phosphorylation of IRS2 and phosphorylation of AktSer473. Hepatic AMPKThr172 phosphorylation was reduced in KTD. In conclusion, KTD impairs energy and glucose homeostasis by exacerbating insulin resistance and attenuating hypothalamic leptin signaling in non-obese type 2 diabetic rats. These changes are not associated with increased serum ketone levels.
Decreased expression level of apoptosis-related genes and/or proteins in skeletal muscles, but not in hearts, of growth hormone receptor knockout mice
Authors: Gesing Adam , Masternak Michal M , Wang Feiya , Lewinski Andrzej , Karbownik-Lewinska Malgorzata , Bartke Andrzej
Published: April 2011

The long-lived growth hormone (GH) receptor knockout (GHRKO; KO) mice are GH-resistant due to targeted disruption of the GH receptor (Ghr) gene. Apoptosis is a physiological process in which cells play an active role in their own death and is a normal component of the development and health of multicellular organisms. Aging is associated with the progressive loss of strength of skeletal and heart muscles. Calorie restriction (CR) is a well-known experimental model to delay aging and increase lifespan. The aim of the study was to examine the expression of the following apoptosis-related genes: caspase-3, caspase-9, caspase-8, bax, bcl-2, Smac/DIABLO, p53 and cytochrome c1 (cyc1) in the skeletal muscles and hearts of female normal and GHRKO mice, fed ad libitum or subjected to 40% CR for six months, starting at two months of age. Moreover, skeletal muscle caspase-3, caspase-9, caspase-8, bax, bcl-2, Smac/DIABLO, Apaf-1, bad, phospho-bad (pbad), phospho-p53 and cytochrome c (cyc) protein expression levels were assessed. Expression of caspase-3, caspase-9, bax and Smac/DIABLO genes and proteins was decreased in GHRKO's skeletal muscles. The Apaf-1 protein expression also was diminished in this tissue. In contrast, bcl-2 and pbad protein levels were increased in skeletal muscles in knockouts. No changes were demonstrated for the examined genes' expression in GHRKO's hearts except for the increased level of cyc1 mRNA. CR did not alter the expression of the examined genes and proteins in skeletal muscles of knockouts versus normal (N) mice. In heart homogenates, CR increased caspase-3 mRNA level as compared with ad libitum mice. Decreased expression of certain proapoptotic genes and/or proteins may constitute the potential mechanism of prolonged longevity in GHRKO mice, protecting these animals from aging; this potential beneficial mechanism is not affected by CR.
Inhibition of DNA synthesis in mouse epidermis by topical imiquimod is dependent on opioid receptors
Authors: McLaughlin Patricia J , Rogosnitzky Moshe , Zagon Ian S
Published: April 2011

The imidazoquinolines are immune response modifiers that have potent antiviral and antitumor properties. The mechanism by which they exert their effects on cell replication has been investigated in vitro and is related to the upregulation of the opioid growth factor receptor (OGFr) and modulation of opioid growth factor (OGF; [Met5]-enkephalin). The OGF–OGFr axis regulates cell proliferative events through a cyclin-dependent kinase inhibitory pathway. The present study examined the mechanism whereby imiquimod repressed cell proliferation in vivo. Using a nude mouse model that has a compromised T-cell immune system, as well as C57BL/6 mice with an intact immune system, the effects of topical imiquimod (Aldara®) on DNA synthesis of basal epithelial cells in skin were examined. Imiquimod's effects on DNA synthesis were detected 24 h after application, and could be observed for one week after a single treatment. The magnitude of change in DNA synthesis following imiquimod was similar for one, three or six applications. Naloxone, an opioid antagonist, blocked the inhibitory effect of imiquimod. Imiquimod in combination with OGF or a low dose of naltrexone (LDN; known to upregulate the OGF–OGFr axis) had no greater inhibitory response on DNA synthesis than either OGF or LDN alone. Both OGF and OGFr were upregulated in basal epithelium after imiquimod treatment. Both nude and C57BL/6 mice exhibited the same repressive action of imiquimod on epithelial DNA synthesis. Imiquimod was neither an opioid agonist nor antagonist using nociceptive testing, and did not induce apoptosis or necrosis. Exposure to imiquimod was found to depress DNA synthesis in cells located in distant epithelium from day 3 and lasted until day 5. These results suggest that the target of imiquimod on DNA synthesis is dependent on an opioid receptor-mediated pathway, and infers that imiquimod is reliant on the OGF–OGFr axis for modulating cell proliferation.
Treatment and risk factor analysis of hypoglycemia in diabetic rhesus monkeys
Authors: He Sirong , Chen Younan , Wei Lingling , Jin Xi , Zeng Li , Ren Yan , Zhang Jie , Wang Li , Li Hongxia , Lu Yanrong , Cheng Jingqiu
Published: April 2011

In order to anticipate and promptly treat hypoglycemia in diabetic monkeys treated with insulin or other glucose-lowering drugs, the relationships between the incidence and symptoms of hypoglycemia in these animals, and many factors involved in model development and sustainment were analyzed. Different procedures were performed on 22 monkeys for the induction of diabetes. The monkey models were evaluated by blood glucose, insulin, C-peptide levels and intravenous glucose tolerance tests. A glucose treatment program for the diabetic monkeys was administered and laboratory tests were regularly performed. A standard procedure of hypoglycemia treatment was established and the risk factors of hypoglycemia were analyzed by a logistic regression model. Furthermore, the relationships between the four methods of diabetes induction, renal function, glycemic control and hypoglycemia were studied using one-way analysis of variance and t-test. We found that the hypoglycemic conditions of diabetic monkeys were improved rapidly by our treatment. The statistical analysis suggested that the modeling methods, renal function and glycemic control were related to the incidence of hypoglycemia. In detail, the progress of diabetes, effects of glycemic control and, particularly, the severity of the hypoglycemia differed according to the induction strategy used. The models induced by partial pancreatectomy with low-dose streptozotocin were not prone to hypoglycemia and their glycemic controls were stable. However, the models induced by total pancreatectomy were more vulnerable to severe hypoglycemia and their glycemic controls were the most unstable. Moreover, the levels of blood creatinine and triglyceride increased after the development of diabetes, which was related to the occurrence of hypoglycemia. In conclusion, we suggested that total pancreatectomy and renal impairment are two important risk factors for hypoglycemia in diabetic monkeys. More attention should be paid to daily care of diabetic monkeys, particularly monitoring and protecting their renal function.
Activation of peroxisome-proliferator-activated receptors α and γ mediates remote ischemic preconditioning against myocardial infarction in vivo
Authors: Lotz Christopher , Lazariotto Maria , Redel Andreas , Smul Thorsten M , Stumpner Jan , Blomeyer Christoph , Tischer-Zeitz Tobias , Schmidt Johannes , Pociej Joanna , Roewer Norbert , Kehl Franz , Lange Markus
Published: April 2011

Remote ischemic preconditioning (remote IPC) elicits a protective cardiac phenotype against myocardial ischemic injury. The remote stimulus has been hypothesized to act on major signaling pathways; however, its molecular targets remain largely undefined. We hypothesized that remote IPC exerts its effects by activating the peroxisome-proliferator-activated receptors (PPARs) α and γ, which have been previously implicated in cardioprotective signaling. Male New Zealand white rabbits ( n = 78) were subjected to a 30-min coronary artery occlusion followed by three hours of reperfusion. Three cycles of remote IPC consisting of 10-min renal ischemia/reperfusion were performed. The animals either received the PPAR α-antagonist GW6471 or the PPAR γ-antagonist GW9662 alone or combined with remote IPC. Infarct size was determined gravimetrically. Tissue levels of 15d-prostaglandin J2 (15d-PGJ2), as well as the PPAR DNA binding were measured using specific assays. Reverse transcriptase polymerase chain reaction was used to analyze changes in endothelial nitric oxide synthase or inducible nitric oxide synthase (iNOS) mRNA expression in relative quantity (RQ). Data are mean ± SD. As a result, remote IPC significantly reduced the myocardial infarct size (42.2 ± 4.9%* versus 61 ± 1.9%), accompanied by an increased PPAR DNA-binding (189.6 ± 19.8RLU* versus 44.4 ± 9RLU), increased iNOS expression (3.5 ± 1RQ* versus 1RQ), as well as 15d-PGJ2 levels (179.7 ± 7.9 pg/mL* versus 127.9 ± 7.6 pg/mL). The protective response elicited by remote IPC, as well as the accompanying molecular changes were abolished by inhibiting PPAR α (56.8 ± 4.7%; 61.1 ± 14.2RLU; and 1.91 ± 0.96RQ, respectively) or PPAR γ (57.4 ± 3.3%; 52.7 ± 16.9RLU; and 1.54 ± 0.25RQ, respectively). (*Significantly different from control P < 0.05). In conclusion, the obtained results indicate that both PPAR α and PPAR γ play an essential role in remote IPC against myocardial infarction, impinging on the transcriptional control of iNOS expression.
Biocompatibility study of glycofurol in rat brains
Authors: Boongird Atthaporn , Nasongkla Norased , Hongeng Suradej , Sukdawong Nongyao , Sa-Nguanruang Waridtha , Larbcharoensub Noppadol
Published: April 2011

Glycofurol (GF) has been used clinically as a solvent for parenteral drug delivery systems. However, the application and toxicity of GF in the brain have not been reported. This study was carried out to assess the systemic and neurologic reactions of GF in rats upon intracranial injection. Hematological and neuropathological assessments of rats were performed during the acute, subacute and chronic period after the injection. Injection of the GF solution (GF 25 μL + PBS 25 μL) into the brain cortex showed that it did not cause any deaths or clinical neurobehavioral abnormalities. At the same volume as phosphate-buffered saline (PBS) injection, it had mild effects on all hematological data and histopathology of brain tissues. Nevertheless, histomorphologic assessments of the brain tissues treated with PBS 70 μL revealed different tissue responses compared with those of 70 μL GF solution (30 μL + PBS 40 μL) where tissues around the administration site showed elevated polymorphonuclear leukocytes, macrophages and gliosis. These results demonstrated that the GF solution (GF 25 μL + PBS 25 μL) administration was well tolerated and caused minor inflammatory responses of cerebral cortex. This suggests possibilities of GF for drug delivery systems in the brain parenchymal tissues.
Functional inhibition of the human middle temporal cortex affects non-visual motion perception: a repetitive transcranial magnetic stimulation study during tactile speed discrimination
Authors: Ricciardi Emiliano , Basso Demis , Sani Lorenzo , Bonino Daniela , Vecchi Tomaso , Pietrini Pietro , Miniussi Carlo
Published: April 2011

The visual motion-responsive middle temporal complex (hMT+) is activated during tactile and aural motion discrimination in both sighted and congenitally blind individuals, suggesting a supramodal organization of this area. Specifically, non-visual motion processing has been found to activate the more anterior portion of the hMT+. In the present study, repetitive transcranial magnetic stimulation (rTMS) was used to determine whether this more anterior portion of hMT+ truly plays a functional role in tactile motion processing. Sixteen blindfolded, young, healthy volunteers were asked to detect changes in the rotation velocity of a random Braille-like dot pattern by using the index or middle finger of their right hand. rTMS was applied for 600 ms (10 Hz, 110% motor threshold), 200 ms after the stimulus onset with a figure-of-eight coil over either the anterior portion of hMT+ or a midline parieto-occipital site (as a control). Accuracy and reaction times were significantly impaired only when TMS was applied on hMT+, but not on the control area. These results indicate that the recruitment of hMT+ is necessary for tactile motion processing, and thus corroborate the hypothesis of a ‘supramodal’ functional organization for this sensory motion processing area.
Over-expressing transient receptor potential cation channel 6 in podocytes induces cytoskeleton rearrangement through increases of intracellular Ca2+ and RhoA activation
Authors: Jiang Lina , Ding Jie , Tsai Haojan , Li Lin , Feng Quancheng , Miao Jing , Fan Qingfeng
Published: April 2011

Transient receptor potential cation channel 6 (TRPC6) is one of the key molecules for filtration barrier function of podocytes. Over-expression of TRPC6 in podocytes is frequently found in acquired or inherited proteinuric kidney diseases, and animal model over-expression of TRPC6 may lead to proteinuria. To investigate the impact of TRPC6 over-expression in podocytes on its function and its relation to proteinuria in kidney diseases, we over-expressed TRPC6 in mouse podocytes by transient transfection of TRPC6 cDNA plasmid, and observed their changes in foot processes, intracellular F-actin distribution, nephrin and synaptopodin expression, electrophysiology, RhoA activity and intracellular Ca2+. In podocytes over-expressing TRPC6, cell processes were reduced remarkably in association with the derangement of cytoskeleton demonstrated by the abnormal distribution of intracellular F-actin. These cells also displayed a higher increase of intracellular Ca2+ ion to the TRPC6 agonist 1-oleoyl-acetyl- sn-glycerol and a higher current in the patch-clamp experiment, down-regulation of nephrin and synaptopodin expression and increase of activated RhoA. These changes could be rescued by the treatment of the cells with U73122 to block TRPC6 channel or BAPTA-AM to chelate intracellular Ca2+ ion. Additionally, the podocytes over-expressing TRPC6 treated with RhoA inhibitor Y-27632 showed an improvement in F-actin arrangement in the cells and increase of nephrin and synaptopodin expression. From these results, we therefore propose that over-expression of TRPC6 in podocytes may be one of the fundamental changes relating to the dysfunction of the slit diaphragm and proteinuria. Podocytes over-expressing TRPC6 may lead to higher intracellular Ca2+ ion concentration in the presence of stimuli. The increase of intracellular Ca2+ down-regulates the expression of two important molecules, nephrin on slit diaphragm and synaptopodin in cytoskeleton, and stimulates RhoA activity, which in turn causes F-actin derangement and the decrease of foot processes.
Shape-specific polymeric nanomedicine: emerging opportunities and challenges
Authors: Tao Li , Hu Walter , Liu Yaling , Huang Gang , Sumer Baran D , Gao Jinming
Published: April 2011

Size and shape are fundamental properties of micro/nanoparticles that are critically important for nanomedicine applications. Extensive studies have revealed the effect of particle size on spherical particles with respect to circulation, extravasation and distribution in vivo. In contrast, the importance of particle shape has only recently begun to emerge. For example, cylindrically-shaped filomicelles (diameter 22–60 nm, length 8–18 μm) have shown persistent blood circulation for up to one week after intravenous injection, much longer than their spherical counterparts. Disc-shaped nanoparticles have demonstrated higher in vivo targeting specificity to endothelial cells expressing intercellular adhesion molecule receptors in mice than spherical particles of similar size. In this Minireview, we will discuss the recent advances in the fabrication of shape-specific nanoparticles and their unique biological and pharmacological properties. Computational models are presented to provide mechanistic understanding of the shape effects on cell targeting under flow conditions. Shape-specific nanoparticles have the potential to significantly improve the performance of nanomedicine in diagnostic imaging and targeted drug delivery applications.
Realgar, cinnabar and An-Gong-Niu-Huang Wan are much less chronically nephrotoxic than common arsenicals and mercurials
Authors: Lu Yuan-Fu , Wu Qin , Yan Jun-Wen , Shi Jing-Zhen , Liu Jie , Shi Jing-Shan
Published: April 2011

Realgar (As4S4) and cinnabar (HgS) are frequently included in traditional Chinese medicines and Indian Ayurvedic medicines. Both As and Hg are well known for toxic effects, and their safety is of concern. The aim of this study was to compare chronic nephrotoxicity of An-Gong-Niu-Huang Wan (AGNH), realgar and cinnabar with common arsenicals and mercurials. Mice were orally administrated with AGNH (3 g/kg, 6-fold of clinical dose), cinnabar (0.3 g/kg, amount in AGNH) and realgar (0.3 g/kg, amount in AGNH), HgCl2 (0.118 mmol/kg, 1/10 of cinnabar), MeHg (0.012 mmol/kg, 1/100 of cinnabar), NaAsO2 (As3+ 0.028 mmol/kg, 1/100 of realgar) or Na2HAsO4 (As5+ 0.056 mmol/kg, 1/50 of realgar), daily for six weeks, and nephrotoxicity was examined. Animal body weights were decreased by MeHg and HgCl2. Blood urea nitrogen and creatinine levels were elevated by MeHg. Renal pathology was severe in the MeHg and HgCl2 groups, moderate in the arsenite, arsenate and realgar groups and mild in the cinnabar and AGNH groups. Renal Hg accumulation in the MeHg and HgCl2 groups was 50–200 folds higher than the cinnabar group. Expressions of metallothionein-1 and heme oxygenase-1, biomarkers for metal toxicity, were increased 2–5 folds by arsenite, arsenate, MeHg and HgCl2, but not by realgar, cinnabar and AGNH. The chemokine and glutathione- S transferase- α4, markers for inflammation, were also increased by MeHg and HgCl2. Expressions of cell adhesion gene S100a9 and E-cadherin were altered by HgCl2, arsenite and realgar. Taken together, chemical forms of mercury and arsenic are major determinants in their disposition and toxicity.
Glycosylphosphatidylinositol-linked ceruloplasmin is expressed in multiple rodent organs and is lower following dietary copper deficiency
Authors: Mostad Elise J , Prohaska Joseph R
Published: April 2011

Ceruloplasmin (Cp), a multicopper ferroxidase, is expressed as both a secreted (sCp) plasma enzyme from the liver and a membrane-bound glycosylphosphatidylinositol-anchored (GPI-Cp) splice variant protein. Cp is thought to be essential for iron mobilization as selective iron overload occurs in aceruloplasminemia in humans and in Cp null mice. Dietary copper-deficient (CuD) rodents have near total loss of Cp activity, severe loss of Cp protein and develop anemia. Hepatic iron augmentation is often observed, suggesting that loss of Cp function may be correlated with anemia. The impact of CuD treatment on GPI-Cp has not previously been evaluated. Our hypothesis was that CuD rodents would have lower levels of GPI-Cp and this would correlate with higher tissue iron retention. In these studies, GPI-Cp was detected in purified membranes of multiple organs of rats and mice but not Cp −/− mice. Immunoreactive Cp protein was released with phosphatidylinositol phospholipase C treatment and expressed ferroxidase activity. Following perinatal and postnatal copper restriction, GPI-Cp was markedly lower in the spleen and modestly lower in the liver of CuD rats and mice, when compared with copper-adequate (CuA) rodents. However, spleen non-heme iron (NHI) was lower in CuD than CuA rats, and not different in CuD mice. Hepatic iron was higher only in CuD mice. Spleen and liver membranes of CuD rats expressed augmented levels of ferroportin, the iron efflux transporter, which may explain lower NHI content in the spleen of CuD rats despite a greater than 50% lower level of the multicopper ferroxidase GPI-Cp. Spleen and liver levels of GPI-Cp mRNA were not impacted in CuD rats, suggesting that turnover rather than biosynthesis may explain the lower steady-state levels of GPI-Cp following dietary copper restriction. Lower GPI-Cp did not correlate with tissue iron retention and thus the role, if any, of Cp in anemia of copper deficiency is unknown.
Pro-inflammatory effects of the aqueous extract of Echinometra lucunter sea urchin spines
Authors: Sciani Juliana Mozer , Zychar Bianca Cestari , Gonçalves Luis Roberto de Camargo , Nogueira Thiago de Oliveira , Giorgi Renata , Pimenta Daniel Carvalho
Published: April 2011

The sea urchin, Echinometra lucunter, can be found along the Western Central Atlantic shores. In Brazil, it is responsible by circa 50% of the accidents caused by marine animals. The symptoms usually surpass trauma and may be pathologically varied and last differently, ranging from spontaneous healing in a few days, to painful consequences lasting for weeks. In this work, we have mimicked the sea urchin accident by administering an aqueous extract of the spine into mice and rats and evaluated the pathophysiological developments. Our data clearly indicate that the sea urchin accident is indeed a pro-inflammatory event, triggered by toxins present in the spine that can cause edema and alteration in the leukocyte–endothelial interaction. Moreover, the spine extract was shown to exhibit a hyperalgesic effect. The extract is rich in proteins, as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but also contains other molecules that can be analyzed by reversed phase high-performance liquid chromatography. Altogether, these effects corroborate that an E. lucunter encounter is an accident and not an incident, as frequently reported by the victims.
Paradoxical increase in liver ketogenesis during long-term insulin-induced hypoglycemia in diabetic rats
Authors: Schiavon Fabiana P M , Gazola Vilma A F G , Furlan Maria M D P , Barrena Helenton C , Bazotte Roberto B
Published: April 2011

It is well established that insulin inhibits liver ketogenesis. However, during insulin-induced hypoglycemia (IIH) the release of counterregulatory hormones could overcome the insulin effect on ketogenesis. To clarify this question the ketogenic activity in livers from alloxan-diabetic rats submitted to long-term IIH was investigated. Moreover, liver glycogenolysis, gluconeogensis, ureagenesis and the production of l-lactate were measured, and its correlation with blood levels of ketone bodies (KB), l-lactate, glucose, urea and ammonia was investigated. For this purpose, overnight fasted alloxan-diabetic rats (DBT group) were compared with control non-diabetic rats (NDBT group). Long-term IIH was obtained with an intraperitoneal injection of Detemir insulin (1 U/kg), and KB, glucose, l-lactate, ammonia and urea were evaluated at 0, 2, 4, 6, 8 or 10 h after insulin injection. Because IIH was well established two hours after insulin injection this time was used for liver perfusion experiments. The administration of Detemir insulin decreased ( P < 0.05) blood KB and glucose levels, but there was an increase in the blood l-lactate levels and a rebound increase in blood KB during the glucose recovery phase of IIH. In agreement with these results, the capacity to produce KB from octanoate was increased in the livers of DBT rats. Moreover, the elevated blood l-lactate levels in DBT rats could be attributed to the higher ( P < 0.05) glycogenolysis when part of glucose from glycogenolysis enters glycolysis, producing l-lactate. In contrast, except glycerol, gluconeogenesis was negligible in the livers of DBT rats. Therefore, during long-term IIH the higher liver ketogenic capacity of DBT rats increased the risk of hyperketonemia. In addition, in spite of the fact that the insulin injection decreased blood KB, there was a risk of worsening lactic acidosis.
microRNAs in hypertrophy and heart failure
Authors: Latronico Michael V G , Condorelli Gianluigi
Published: April 2011

microRNAs (miRNAs) are highly conserved, short non-coding RNAs that have become established as having an important role in regulatory networks. We review here the progress made in our understanding of heart-related microRNAs over the last two years, focusing mainly on hypertrophic growth and heart failure.
Revival of the identification of cytotoxic T-lymphocyte epitopes for immunological diagnosis, therapy and vaccine development
Authors: Liu Jun , Zhang Shihong , Tan Shuguang , Zheng Beiwen , Gao George F
Published: April 2011

Immunogenic T-cell epitopes have a central role in the cellular immunity against pathogens and tumors. However, in the early stage of cellular immunity studies, it was complicated and time-consuming to identify and characterize T-cell epitopes. Currently, the epitope screening is experiencing renewed enthusiasm due to advances in novel techniques and theories. Moreover, the application of T-cell epitope-based diagnoses for tuberculosis and new data on epitope-based vaccine development have also revived the field. There is a growing knowledge on the emphasis of epitope-stimulated T-cell immune responses in the elimination of pathogens and tumors. In this review, we outline the significance of the identification and characterization of T-cell epitopes. We also summarize the methods and strategies for epitope definition and, more importantly, address the relevance of cytotoxic T-lymphocyte epitopes to clinical diagnoses, therapy and vaccine development.