bluuebunny/crossref_metadata_2025 huggingface dataset:
Stimulation of Sigma-1 receptor by dehydroepiandrosterone ameliorates hypertension-induced kidney hypertrophy in ovariectomized rats
Authors: Bhuiyan Shenuarin , Fukunaga Kohji
Published: May 2010

The incidence of chronic renal disease in women increases with aging, especially after menopause, suggesting that loss of sex hormones contributes to the development and progression of renal diseases. Recent studies revealed that decreased dehydroepiandrosterone (DHEA) levels are associated with endothelial dysfunction, renal injury and increased cardiovascular mortality in postmenopausal women. We here investigate the role of DHEA, also known as Sigma-1 receptor (Sigma-1R) agonist, on kidney injury induced by pressure overload (PO) after ovariectomy (OVX) and defined mechanisms underlying its protective action. Wistar rats subjected to bilateral OVX were further treated with abdominal aortic stenosis between the right and left renal arteries. DHEA (15 and 30 mg/kg) was administered orally once a day for 14 days starting from two weeks after aortic banding. Time course study indicated that the right kidney (RK) weight-to-body weight (BW) ratio increases time-dependently from one to four weeks along with increased mean arterial blood pressure (MABP) after banding in the abdominal aorta with no change in the left kidney (LK) weight-to-BW ratio. Similarly, we found significant time-dependent decrease in Sigma-1R expression in the RK with no changes in the LK. Administration of the Sigma-1R agonist, DHEA, significantly inhibited hypertension-induced increases in the RKW-to-BW ratio and increased expression of Sigma-1R in the RK. DHEA also attenuated PO-induced disturbance of heart rate and MABP. DHEA administration significantly restored PO-induced impaired endothelial nitric oxide synthase (eNOS) activity with concomitant increased phosphorylation of eNOS (Ser1179) and Akt activity with increased phosphorylation at Ser 473 and at Thr 308 in the RK. We here documented, for the first time, the potential role of Sigma-1R to protect the kidney from PO-induced injury in ovariectomized rats. DHEA administration protects hypertension-induced kidney injury via upregulation of Sigma-1R and stimulation of Akt–eNOS signaling in ovariectomized rats.
Isolation and characterization of multipotent progenitor cells from the human fetal aorta wall
Authors: Fang Baijun , Li Yufu , Song Yongping , Li Ning
Published: May 2010

Recent evidence indicates that vascular progenitor cells may be the source of smooth muscle cells (SMCs) and endothelial cells (ECs). In the present study we isolated CD105+, CD34– and fetal liver kinase+ (Flk1+) cells from the human fetal arterial wall and demonstrated that they were vascular progenitors for both ECs and SMCs. In vitro, these cells cultured with vascular endothelial growth factor could differentiate into cells that expressed endothelial markers. Meanwhile, cells cultured with platelet-derived growth factor-BB could differentiate into cells that expressed smooth muscle markers. When transplanted into NOD/SCID mice, they contributed to neoangiogenesis in vivo during wound healing. These cells could also differentiate into osteogenic and adipogenic lineages in vitro. Hence multipotent vascular progenitor cells do exist in the arterial wall and they may have implications in the physical and pathological conditions of the vessel. Because these cells can be expanded in culture without obvious senescence for more than 30 population doublings, they may be an important source of ECs for cellular pro- or anti-angiogenic therapies.
Central, peripheral and resistance arterial reactivity: fluctuates during the phases of the menstrual cycle
Authors: Adkisson Eric J , Casey Darren P , Beck Darren T , Gurovich Alvaro N , Martin Jeffery S , Braith Randy W
Published: May 2010

The purpose of this study was to document the temporal changes in vascular reactivity occurring simultaneously in central, peripheral and microvascular resistance arteries in the same cohort of women during the normal menstrual cycle. Twenty-three ( n = 23) women (mean age (±SD) = 19 ± 1 y) were tested during four phases of a normal menstrual cycle. Delineation of the four phases occurred as follows: (1) the early follicular phase; (2) the late follicular (LF) phase; (3) the early luteal (EL) phase; and (4) the late luteal phase. Non-invasive measurement of central hemodynamics and peripheral artery pulse wave velocity (PWV) were performed using applanation tonometry. Measurement of peripheral endothelial function was determined by flow-mediated dilation (FMD) testing in the brachial artery and venous occlusion plethysmography in the forearm and calf resistance arteries. Additionally, plasma NO x and 17 β-estradiol (E) concentrations were measured. Both central (aortic) and peripheral blood pressure (BP) were lowest ( P < 0.05) during the LF phase and BP reduction was sustained ( P < 0.05) into the EL phase. The timing and amplitude of the reflected pressure wave were attenuated only during the LF phase ( P < 0.05). No temporal changes were observed in either central (carotid-femoral) or peripheral PWV (femoral-dorsalis pedis, carotid-radial). Peak forearm and calf blood flow during reactive hyperemia were greatest in LF. Brachial FMD was greatest during the LF phase ( P < 0.05). Plasma E and NO x concentrations were highest during the LF phase ( P < 0.05). Young premenopausal women experienced an overwhelming pattern of reduced BP and increased systemic vascular reactivity during the LF phase prior to ovulation.
Antiangiogenic therapy in lung cancer: focus on vascular endothelial growth factor pathway
Authors: Korpanty Grzegorz , Smyth Elizabeth , Sullivan Laura A , Brekken Rolf A , Carney Desmond N
Published: May 2010

Lung cancer (LC) is a leading cause of death worldwide. Recent advances in chemotherapeutic agents have not yielded any significant improvement in the prognosis of patients with LC. The five-year survival rate for all combined disease stages remains about 15%. For this reason, new therapies such as those that inhibit tumor angiogenesis or block activity of growth factor receptors are of special interest in this group of patients. In this review we will summarize the most recent clinical data on biologic therapies that inhibit tumor angiogenesis in LC, focusing on those that are most clinically relevant.
Role of potassium channels in coronary vasodilation
Authors: Dick Gregory M , Tune Johnathan D
Published: May 2010

K+channels in coronary arterial smooth muscle cells (CASMC) determine the resting membrane potential ( Em) and serve as targets of endogenous and therapeutic vasodilators. Emin CASMC is in the voltage range for activation of L-type Ca2+channels; therefore, when K+channel activity changes, Ca2+influx and arterial tone change. This is why both Ca2+channel blockers and K+channel openers have such profound effects on coronary blood flow; the former directly inhibits Ca2+influx through L-type Ca2+channels, while the latter indirectly inhibits Ca2+influx by hyperpolarizing Emand reducing Ca2+channel activity. K+channels in CASMC play important roles in vasodilation to endothelial, ischemic and metabolic stimuli. The purpose of this article is to review the types of K+channels expressed in CASMC, discuss the regulation of their activity by physiological mechanisms and examine impairments related to cardiovascular disease.
Tumor-derived intercellular adhesion molecule-1 mediates tumor-associated leukocyte infiltration in orthotopic pancreatic xenografts
Authors: Roland Christina L , Dineen Sean P , Toombs Jason E , Carbon Juliet G , Smith C Wayne , Brekken Rolf A , Barnett Carlton C
Published: May 2010

Tumor infiltration of immune cells (polymorphonuclear cells [PMNs] and macrophages) was initially thought to be an attempt by the host organism to combat malignancy. It appears, however, that certain subsets of chronically activated immune cells likely promote tumor growth, facilitate tumor cell survival and aid in metastasis. The association between tumor cells and tumor-associated PMNs has been demonstrated in several types of cancer, but the presence of tumor-associated PMNs in pancreatic cancer has not been well studied in vivo. Intercellular adhesion molecule-1 (ICAM-1) functions in cell–cell and cell–extracellular matrix adhesion and has a physiological role in PMN tight adhesion of leukocytes via interaction with the ligands LFA-1 and Mac-1. Increased ICAM-1 expression correlates with poor prognosis in pancreatic cancer. Therefore, the aim of this study was to investigate the function of ICAM-1 and tumor-associated PMNs in pancreatic cancer progression using ICAM-1-null (ICAM-1−/−) mice. We hypothesize that ICAM-1 null mice have decreased pancreatic cancer progression. Surprisingly, there is no significant difference in pancreatic cancer progression in wild-type versus ICAM-1 null mice. Interestingly, we found that tumor-derived ICAM-1 co-localizes with host PMNs at the leading edge of the tumor in ICAM-1 null mice. These results suggest that tumor-derived ICAM-1 is a sufficient ligand for tumor-associated PMNs and may play a role in subsequent tumor growth and metastasis.
Buddleja officinalis suppresses high glucose-induced vascular smooth muscle cell proliferation: role of mitogen-activated protein kinases, nuclear factor-κB and matrix metalloproteinases
Authors: Lee Yun Jung , Kim Jin Sook , Kang Dae Gill , Lee Ho Sub
Published: May 2010

Diabetes mellitus is a well-established risk factor for vascular diseases caused by atherosclerosis. In the development of diabetic atherogenesis, vascular smooth muscle cell proliferation is recognized as a key event. Thus, we aimed to investigate whether an ethanol extract of Buddleja officinalis (EBO) suppresses high glucose-induced proliferation in primary cultured human aortic smooth muscle cells (HASMC). [3H]-thymidine incorporation revealed that incubation of HASMC with a high concentration of glucose (25 mmol/L) increased cell proliferation. The expression levels of cell cycle protein were also increased by treatment with high glucose concentration. Pretreatment of HASMC with EBO significantly attenuated the increase of high glucose-induced cell proliferation as well as p38 mitogen-activated protein kinases (MAPK) and JNK phosphorylation. EBO suppressed high glucose-induced matrix metalloproteinase (MMP)-9 activity in a dose-dependent manner. In addition, EBO suppressed nuclear factor- κB (NF- κB) nuclear translocation and transcriptional activity in high glucose conditions. Taken together, the present data suggest that EBO could suppress high glucose-induced atherosclerotic processes through inhibition of p38, JNK, NF- κB and MMP signal pathways in HASMC.
Dual oxidase, hydrogen peroxide and thyroid diseases
Authors: Ohye Hidemi , Sugawara Masahiro
Published: May 2010

The thyroid gland is a unique endocrine organ that requires hydrogen peroxide (H2O2) for thyroid hormone formation. The molecule for H2O2 production in the thyroid gland has been known as dual oxidase 2 (DUOX2). Recently, NADPH oxidase 4 (NOX4), a homolog of the NOX family, was added as a new intracellular source of reactive oxygen species (ROS) in the human thyroid gland. This review focuses on the recent progress of the DUOX system and its possible contribution to human thyroid diseases. Also, we discuss human thyroid diseases related to abnormal H2O2 generation. The DUOX molecule contains peroxidase-like and NADPH oxidase-like domains. Human thyroid gland also contains DUOX1 that shares 83% similarity with the DUOX2 gene. However, thyroid DUOX1 protein appears to play a minor role in H2O2 production. DUOX proteins require DUOX maturation or activation factors (DUOXA1 or 2) for proper translocation of DUOX from the endoplasmic reticulum to the apical plasma membrane, where H2O2 production takes place. Thyroid cells contain antioxidants to protect cells from the H2O2-mediated oxidative damage. Loss of this balance may result in thyroid cell dysfunction and thyroid diseases. Mutation of either DUOX2 or DUOXA2 gene is a newly recognized cause of hypothyroidism due to insufficient H2O2 production. Papillary thyroid carcinoma, the most common thyroid cancer, is closely linked to the increased ROS production by NOX4. Hashimoto's thyroiditis, a common autoimmune thyroid disease in women, becomes conspicuous when iodide intake increases. This phenomenon may be explained by the abnormality of iodide-induced H2O2 or other ROS in susceptible individuals. Discovery of DUOX proteins and NOX4 provides us with valuable tools for a better understanding of pathophysiology of prevalent thyroid diseases.
Proposal for a role of the Hsp90/Hsp70-based chaperone machinery in making triage decisions when proteins undergo oxidative and toxic damage
Authors: Pratt William B , Morishima Yoshihiro , Peng Hwei-Ming , Osawa Yoichi
Published: May 2010

The Hsp90/Hsp70-based chaperone machinery plays a well-established role in signaling protein function, trafficking and turnover. A number of recent observations also support the notion that Hsp90 and Hsp70 play key roles in the triage of damaged and aberrant proteins for degradation via the ubiquitin-proteasome pathway. In the mid-1990s, it was discovered that Hsp70 is required for ubiquitin-dependent degradation of short-lived and abnormal proteins, and it became clear that inhibition of Hsp90 uniformly leads to the proteasomal degradation of Hsp90 client proteins. Subsequently, CHIP and parkin were shown to be Hsp70-binding ubiquitin E3 ligases that direct ubiquitin-charged E2 enzymes to the Hsp70-bound client protein. Stabilization by Hsp90 reflects the interaction of the chaperone with the ligand binding cleft of the client protein. These hydrophobic clefts must be open to allow passage of ligands to binding sites in the protein interior, and they are inherent sites of conformational instability. Hsp90 stabilizes the open state of the cleft and prevents Hsp70-dependent ubiquitination. In the model we propose here, progressive oxidative events result in cleft opening as the initial step in protein unfolding, and as long as Hsp90 can interact to stabilize the cleft, it will buffer the effect of oxidative damage. When cleft opening is such that Hsp90 can no longer interact, Hsp70-dependent ubiquitination occurs. We summarize evidence that Hsp90 interacts very dynamically with a variety of proteins that are not classic Hsp90 clients, and we show that this dynamic cycling of Hsp90 with nitric oxide synthase protects against CHIP-mediated ubiquitination. Scientific interest to date has focused on stringent regulation of the classic client proteins, which have metastable clefts and are inherently short lived. But, the recognition that Hsp90 cycles dynamically with longer lived proteins with more stable clefts may permit extension of the triage model to the quality control of damaged proteins in general.
Histone modification-mediated CYP2E1 gene expression and apoptosis of HepG2 cells
Authors: Yang Hui , Nie Yuqiang , Li Yuyuan , Wan Yu-Jui Yvonne
Published: May 2010

The incidence of hepatocellular carcinoma is rising due to alcohol drinking, hepatitis C viral infection and metabolic syndrome. Differential expression of CYP2E1 may play a pleiotropic role in the multistep process of liver carcinogenesis. Considerable attention has focused on the antitumor effect of trichostatin A (TSA) as well as CYP2E1 expression-induced apoptosis of cancer cells. However, very few studies have examined the mechanisms by which TSA has an antitumor effect and its association to CYP2E1 expression. The current study examined the action of TSA on CYP2E1 expression and the role of CYP2E1 in inducing apoptosis of HepG2 cells. Our data showed that TSA selectively induced CYP2E1 in four studied human hepatocellular carcinoma (HCC) cell lines (Huh7, PLC/PRF/5, Hep3B and HepG2), but not in normal primary human hepatocytes. TSA-mediated up-regulation of CYP2E1 expression was associated with histone H3 acetylation and the recruitment of HNF-1 and HNF-3 β to the CYP2E1 promoter in HepG2 cells. siRNA-mediated knockdown experiments showed that TSA-induced caspase-3 cleavage was decreased due to reduced expression of CYP2E1 in HepG2 cells. Moreover, down-regulation of CYP2E1 was accompanied by decreased production of mitochondrial reactive oxygen species. These results suggest that histone modification is involved in CYP2E1 gene expression and that CYP2E1-dependent mitochondrial oxidative stress plays a role in TSA-induced apoptosis.
Apoptosis caused by an inhibitor of NO production in the decidua of rat from mid-gestation
Authors: Suzuki Takehito , Nagamatsu Chiaki , Kushima Takahiro , Miyakoshi Ryu , Tanaka Kazuaki , Morita Hidetoshi , Sakaue Motoharu , Takizawa Tatsuya
Published: May 2010

We previously reported that nitric oxide (NO) is first detected in the uterus of a pregnant rat on gestational day 13.5 (GD13.5) and that NO levels peak on GD17.5. In addition, NO production in the uterus is mainly derived from the decidua and not the myometrium. The aim of the present study was to reveal the role of NO that peaked on GD17.5 of gestation in the decidua. To inhibit NO production, pregnant rats were continuously administered by an nitric oxide synthase inhibitor, NG-nitro-l-arginine-methyl ester (l-NAME) for 48 h. In the control group, saline was infused instead of l-NAME. After treatment, the decidua were obtained from GD13.5, GD17.5 and GD21.5 rats. Apoptosis and activated caspase-3-positive cells were observed by transferase-mediated dUTP nick-end labeling (TUNEL) assay and immunohistochemistry, respectively. The caspase-3 enzyme activity was also measured in the cell lysate from the decidua. The numbers of TUNEL-positive cells and activated caspase-3-positive cells each increased and the amount of caspase-3 activity also increased significantly in rats on GD17.5 than in rats in the control group, but no changes were observed in rats on GD13.5 and GD21.5. Furthermore, enzyme activity regarding the initiator caspases, caspase-8 and -9, upstream factors for caspase-3 in the caspase cascade, was measured simultaneously on GD17.5 under the same treatment. Caspase-8 and -9 enzyme activities increased significantly in the control group; an increment of caspase-8 activity was especially prominent. The present results indicate that an inhibitor of NO production caused apoptosis through typical apoptotic signals in the decidua on GD17.5, suggesting that an NO peak in the decidua is essential to cell survival and the maintenance of uterine formation.
Tissue protective effect of xanthine oxidase inhibitor, polymer conjugate of (styrene–maleic acid copolymer) and (4-amino-6-hydroxypyrazolo[3,4-d]pyrimidine), on hepatic ischemia–reperfusion injury
Authors: Fang Jun , Seki Takahiro , Qin Haibo , Bharate Gahininath Y , Iyer Arun K , Maeda Hiroshi
Published: May 2010

The detrimental role of superoxide anion (O2−) has been well documented in the pathogenesis of ischemia–reperfusion (I/R) injury. Our and other studies suggested that one critical source of O2−generation may be xanthine oxidase (XO). We thus hypothesized that I/R injury could be protected by inhibiting XO activity, which would reduce the amount of O2−and hence reduce pathogenic consequences. Among various XO inhibitors, we previously found 4-amino-6-hydroxypyrazolo[3,4- d]pyrimidine (AHPP) exhibited potent XO inhibitory activity. Here, we report that the covalent conjugate of AHPP with amphipathic styrene–maleic acid copolymer (SMA-AHPP) showed protective effect against I/R-induced injury in a rat hepatic I/R model. Liver ischemia was induced by occluding both the portal vein and the hepatic artery for 30 min, and followed by reperfusion. SMA-AHPP was administered via the tail vein two hours before ischemia was initiated. A remarkable increase of liver enzymes in plasma (aspartate aminotransferase, AST; alanine aminotransferase, ALT and lactate dehydrogenase, LDH) was detected three hours after reperfusion, whereas prior injection of SMA-AHPP greatly suppressed this increase of AST, ALT and LDH. Moreover, induction of inflammatory cytokines, i.e. tumor necrosis factor-alpha (TNF- α), interleukin-12 (IL-12) and monocyte chemotactic protein-1 (MCP-1) by I/R were significantly inhibited by SMA-AHPP treatment. Accordingly, cytotoxic effect or apoptosis in the liver caused by I/R was clearly reduced by SMA-AHPP pretreatment. Furthermore, thiobarbituric acid-reactive substance assay showed a significant decrease of lipid peroxidation in rat liver after the administration of SMA-AHPP, which is parallel with the decreased XO activity after SMA-AHPP treatment, indicating the involvement of reactive oxygen species generated by XO. In addition, SMA-AHPP was found to bind to albumin, thus to exhibit prolonged in vivo (plasma) half-life. These results suggest that SMA-AHPP exerted a potent cytoprotective effect against I/R injury in rat liver, by inhibiting XO activity and the subsequent generation of O2−.
Free zinc ions outside a narrow concentration range are toxic to a variety of cells in vitro
Authors: Bozym Rebecca A , Chimienti Fabrice , Giblin Leonard J , Gross Gunter W , Korichneva Irina , Li Yuan , Libert Sarah , Maret Wolfgang , Parviz Maryam , Frederickson Christopher J , Thompson Richard B
Published: May 2010

The zinc(II) ion has recently been implicated in a number of novel functions and pathologies in loci as diverse as the brain, retina, small intestine, prostate, heart, pancreas, and immune system. Zinc ions are a required nutrient but elevated concentrations are known to kill cells in vitro. Paradoxical observations regarding zinc's effects have appeared frequently in the literature, and often their physiological relevance is unclear. We found that for PC-12, HeLa and HT-29 cell lines as well as primary cultures of cardiac myocytes and neurons in vitro in differing media, approximately 5 nmol/L free zinc (pZn = 8.3, where pZn is defined as – log10 [free Zn2+]) produced apparently healthy cells, but 20-fold higher or (in one case) lower concentrations were usually harmful as judged by multiple criteria. These results indicate that (1) the free zinc ion levels of media should be controlled with a metal ion buffer; (2) adding zinc or strong zinc ligands to an insufficiently buffered medium may lead to unpredictably low or high free zinc levels that are often harmful to cells; and (3) it is generally desirable to measure free zinc ion levels due to the presence of contaminating zinc in many biochemicals and unknown buffering capacity of many media.
Clinopodium gracile inhibits mast cell-mediated allergic inflammation: involvement of calcium and nuclear factor-κB
Authors: Park Seung-Bin , Kim Sang-Hyun , Suk Kyoungho , Lee Hyun-Shik , Kwon Taeg Kyu , Ju Min-Gyu , Jeon Hoon , Kim Dae-Keun , Lim Jong-Pil , Shin Tae-Yong
Published: May 2010

Mast cell-mediated allergic disease is involved in many diseases such as anaphylaxis, rhinitis, asthma and atopic dermatitis. The discovery of drugs for the treatment of allergic disease is an important subject in human health. In this study, we investigated the effect of the water extract of Clinopodium gracile Matsum var. multicaule (WECG) on the mast cell-mediated allergic inflammation and studied the possible mechanism of action. WECG inhibited compound 48/80-induced systemic anaphylaxis and immunoglobulin E-mediated cutaneous anaphylaxis in a dose-dependent manner. WECG dose-dependently reduced histamine release from rat peritoneal mast cells and human mast cells. The inhibitory effect of WECG on histamine release was mediated by the modulation of intracellular calcium. In addition, WECG attenuated the phorbol 12-myristate 13-acetate and calcium ionophore A23187-stimulated gene expression and secretion of proinflammatory cytokines such as tumor necrosis factor- α and interleukin-6 in human mast cells. The inhibitory effect of WECG on these proinflammatory cytokines was nuclear factor- κB (NF- κB) dependent. Our findings provide evidence that WECG inhibits mast cell-derived allergic inflammation and involvement of calcium and NF- κB in these effects.
Rapamycin reduces intrahepatic alpha-1-antitrypsin mutant Z protein polymers and liver injury in a mouse model
Authors: Kaushal Shalesh , Annamali Mani , Blomenkamp Keith , Rudnick David , Halloran Donna , Brunt Elizabeth M , Teckman Jeffrey H
Published: May 2010

Alpha-1-antitrypsin (a1AT) deficiency is caused by homozygosity for the a1AT mutant Z gene and occurs in one in 2000 Americans. The Z mutation confers an abnormal conformation on the a1AT mutant Z protein, resulting in accumulation within the endoplasmic reticulum of hepatocytes and chronic liver injury. Autophagy is one of several proteolytic mechanisms activated to cope with this hepatocellular protein burden, and is likely important in disposal of the unique polymerized conformation of the a1AT mutant Z protein, which is thought to be especially injurious to the cell. Recent data indicate that rapamycin may more efficiently upregulate autophagy when given in weekly dose pulses, as compared with a daily regimen. Therefore, we evaluated the effect of rapamycin on PiZ mice, a well-characterized model which recapitulates human a1AT liver disease. Daily dosing had no effect on autophagy, on accumulation of a1AT mutant Z protein or on liver injury. Weekly dosing of rapamycin did increase autophagic activity, as shown by increased numbers of autophagic vacuoles. This was associated with reduction in the intrahepatic accumulation of a1AT mutant Z protein in the polymerized conformation. Markers of hepatocellular injury, including cleavage of caspase 12 and hepatic fibrosis, were also decreased. In conclusion, this is the first report of a successful in vivo method for reduction of intrahepatic a1AT mutant Z polymerized protein. Application of this finding may be therapeutic in patients with a1AT deficiency by reducing the intracellular burden of the polymerized, mutant Z protein and by reducing the progression of liver injury.
Functional characterization of rhesus embryonic stem cell-derived serotonin neurons
Authors: Tokuyama Yukari , Ingram Susan L , Woodward Joy S , Bethea Cynthia L
Published: May 2010

Optimal function of the serotonin system is essential for mental health and its role in psychopathologies is undisputed. Enhancing the ability to study primate serotonin neurons in culture would facilitate understanding of intracellular signaling pathways that mediate the action of drugs and other epigenetic or developmental factors impacting human mental health. We were the first group to report differentiation of the non-human primate rhesus monkey embryonic stem cell (ESC) line 366.4 into cultures of serotonin neurons. In this study, we optimized yield and obtained functional characteristics of the derived serotonin neurons. Sequential treatments of ESC 366.4 during expansion stage with fibroblast growth factor 4 and sonic hedgehog markedly increased the yield of serotonin neurons. These serotonin neurons propagated action potentials and expressed GABA receptors. Also, for the first time we demonstrate that these ESC-derived serotonin neurons exhibit functional high-affinity transporter sites, as well as high-affinity 5HT1Abinding sites, which are essential targets of common psychoactive drugs. Finally, to test the generality of this method, we utilized another rhesus ESC line, ORMES-22, which efficiently differentiated into serotonin neurons. Together, these findings demonstrate the feasibility of our protocol to direct different primate ESC lines to serotonin neurons with physiological characteristics, which makes them a useful in vitro model system.
Prostaglandin E2: the master of endometriosis?
Authors: Wu Meng-Hsing , Lu Chun-Wun , Chuang Pei-Chin , Tsai Shaw-Jenq
Published: May 2010

Endometriosis is the primary cause of infertility in women, with a prevalence rate ranging from 5% to 10%. Women with endometriosis suffer from symptoms such as chronic pelvic pain, dysmenorrhea and dyspareunia, which significantly reduce the quality of life. Endometriosis is a polygenic disease with a complex, multifactorial etiology. The mechanism responsible for the initiation and development of this disease remains largely unknown. Prostaglandin E2(PGE2), a versatile eicosanoid that exerts numerous physiological and pathological functions, has been implicated to play critical roles in the development of endometriosis. A growing body of evidence demonstrates that PGE2regulates many pathophysiological processes including cell proliferation, antiapoptosis, immune suppression and angiogenesis during the development of endometriosis. This review focuses on recent advances in cellular and molecular mechanisms triggered by PGE2that contribute to the pathological processes of endometriosis.
Encapsulated plant extract (Gelsemium sempervirens) poly (lactide-co-glycolide) nanoparticles enhance cellular uptake and increase bioactivity in vitro
Authors: Bhattacharyya Soumya Sundar , Paul Saili , Khuda-Bukhsh Anisur Rahman
Published: May 2010

Ethanolic extract of Gelsemium sempervirens (family: Loganiaceae), henceforth to be called EEGS, is used in various traditional systems of medicine. In homeopathy, EEGS is known as mother tincture of G. sempervirens, which is generally used to treat pain and respiratory ailments. We demonstrated earlier anticancer activity of crude EEGS by in vitro studies on human HeLa cells. To test the hypothesis if nanoparticle-encapsulated extract (now onwards to be called NEEGS) could enhance cellular uptake and thereby improve bioactivity, we formulated nanoparticle encapsulation based on poly (lactide-co-glycolide) (PLGA) and confirmed encapsulation by scanning electron microscopy (SEM) and atomic force microscopy. EEGS was encapsulated with 81.6% efficiency in PLGA biodegradable nanoparticle formulation and F68 (polyoxyethylene-polyoxypropylene) was used as a stabilizer. Dynamic laser light scattering and SEM indicated a particle diameter of 122.6 nm. The zeta potential of the drug-loaded nanoparticles was −14.8 mV. NEEGS was characterized for their biological activities in a skin cancer cell line A375 in vitro. NEEGS exhibited relatively rapid (30 min) and more efficient cellular uptake than their un-encapsulated counterpart (45 min). Analysis of data of apoptosis study using Annexin V-FITC, terminal transferase dUTP nick end labeling assay and DNA ladder revealed that encapsulated EEGS was more potent than their un-encapsulated counterpart in inducing apoptosis of A375 cells. Reverse transcriptase-polymerase chain reaction data of survivin, cyclin-D1, caspase-3, PCNA and p53 also corroborated well to suggest greater potentials of NEEGS as anticancer agents.
Signaling pathways involved in postconditioning-induced cardioprotection of human myocardium, in vitro
Authors: Lemoine Sandrine , Puddu Paolo Emilio , Durand Charline , Lepage Olivier , Babatasi Gérard , Ivascau Calin , Massetti Massimo , Gérard Jean-Louis , Hanouz Jean-Luc
Published: May 2010

We examined the respective role and relationship between protein kinase C (PKC), mitochondrial adenosine triphosphate-sensitive potassium (mitoKATP) channel and p38 mitogen-activated protein kinase (MAPK) in postconditioning of human myocardium, in vitro. Isometrically contracting, isolated human right atrial trabeculae were exposed to 30 min hypoxia and 60 min reoxygenation. Phorbol 12-myristate 13-acetate (a PKC activator), diazoxide (a mitoKATP opener) and anisomycin (a p38 MAPK activator) were superfused in early reoxygenation alone and with calphostin C (a PKC inhibitor), 5-hydroxy-decanoate (5-HD, a mitoKATP channel inhibitor) and SB 202190 (a p38 MAPK inhibitor). Developed force at the end of the 60 min reoxygenation (FoC60) period was compared between groups (mean ± SD). Phorbol 12-myristate 13-acetate (91 ± 4% of baseline), diazoxide (85 ± 5% of baseline) and anisomycin (90 ± 4% of baseline) enhanced the FoC60 as compared with the control group (53 ± 7% of baseline, P < 0.0001). The enhanced FoC60 induced by phorbol 12-myristate 13-acetate was abolished by calphostin C (52 ± 5% of baseline) and 5-HD (56 ± 3% of baseline), but not by SB 202190 (90 ± 8%). The diazoxide-induced recovery of FoC60 was attenuated by 5-HD (55 ± 6% of baseline), but was not modified by calphostin C (87 ± 5% of baseline) and SB 202190 (90 ± 8% of baseline). The anisomycin-induced recovery of FoC60 was abolished by calphostin C (61 ± 9% of baseline) and SB 202190 (52 ± 8% of baseline), but not by 5-HD (88 ± 6% of baseline). In conclusion, PKC activation, opening of mitoKATP channels and p38 MAPK activation in early reoxygenation induced the postconditioning of human myocardium, in vitro. Furthermore, PKC activation was upstream of the opening of mitoKATP channels; p38 MAPK acted on PKC. Therefore, mitoKATP and p38 MAPK seemed to be involved in two independent pathways.
Effects of electro-acupuncture on endothelium-derived endothelin-1 and endothelial nitric oxide synthase of rats with hypoxia-induced pulmonary hypertension
Authors: Pan Peng , Zhang Xueyong , Qian Hua , Shi Weidong , Wang Juan , Bo Yulong , Li Wenzhi
Published: May 2010

Pulmonary hypertension (PH) is characterized by elevated pulmonary artery pressure (PAP), pulmonary vascular remodeling and right ventricular hypertrophy, which are mainly due to endothelial dysfunction. Electro-acupuncture has shown beneficial effects on cardiovascular homeostasis, but little evidence has been obtained on pulmonary effects. The goal of the present study was to investigate whether electro-acupuncture on bladder-13 and -15 points can protect against chronic hypoxia-induced PH by regulating endothelium-derived endothelin (ET)-1 and endothelial nitric oxide synthase (eNOS). Male Wistar rats were exposed to hypoxia to induce PH. Hemodynamic analysis revealed that mean PAP was similar under normoxic conditions. Chronic hypoxia increased mean PAP to 37 ± 3 mmHg, and electro-acupuncture attenuated it to 29 ± 3 mmHg. Absolute right ventricular weight was ameliorated by electro-acupuncture from 0.288 ± 0.048 g to 0.228 ± 0.029 g under hypoxic conditions. Hypoxia-induced right ventricular hypertrophy index decreased from 0.477 ± 0.069 to 0.378 ± 0.053 with electro-acupuncture treatment. Histological examination revealed that hypoxic rats showed increased medial pulmonary artery wall thickness as well as muscularization. However, these alternations by chronic hypoxia were attenuated by electro-acupuncture. There was no difference in eNOS or ET-1 between groups under normoxic conditions. Electro-acupuncture treatment significantly improved the circulating eNOS concentration (365.36 ± 31.51 pg/mL) compared with only hypoxia exposure (247.60 ± 30.64 pg/mL). In lung homogenate, levels of eNOS under hypoxia increased from 684.96 ± 117.90 to 869.86 ± 197.61 pg/mg by electro-acupuncture treatment. Levels of ET-1 changed oppositely to eNOS in response to electro-acupuncture (ET-1 in plasma, 29.44 ± 2.09 versus 20.70 ± 2.37 pg/mL; ET-1 in lung homogenate, 120.51 ± 3.03 versus 110.60 ± 4.04 pg/mg). In conclusion, these results indicated that treatment with electro-acupuncture can protect against hypoxia-induced PH, possibly by regulating the balance of endothelium-derived vasoconstrictors and vasodilators.
Apigenin and naringenin suppress colon carcinogenesis through the aberrant crypt stage in azoxymethane-treated rats
Authors: Leonardi Tety , Vanamala Jairam , Taddeo Stella S , Davidson Laurie A , Murphy Mary E , Patil Bhimanagouda S , Wang Naisyin , Carroll Raymond J , Chapkin Robert S , Lupton Joanne R , Turner Nancy D
Published: May 2010

Epidemiological evidence suggests that a diet abundant in fruits and vegetables may protect against colon cancer. Bioactive compounds, including flavonoids and limonoids, have been shown to possess antiproliferative and antitumorigenic effects in various cancer models. This experiment investigated the effects of four citrus flavonoids and one limonoid mixture at the promotion stage of chemically induced colon cancer in rats. Male Sprague–Dawley rats ( n = 10 rats/group) were randomly allocated to one of six diets formulated to contain 0.1% apigenin, 0.02% naringenin, 0.1% hesperidin, 0.01% nobiletin, 0.035% limonin glucoside/obacunone glucoside mixture or a control diet (0% flavonoid/limonoid). Rats received experimental diets for 10 weeks and were injected with azoxymethane (15 mg/kg) at weeks 3 and 4. Excised colons were evaluated for aberrant crypt foci (ACF) formation, colonocyte proliferation (proliferating cell nuclear antigen assay), apoptosis (terminal deoxynucleotidyl transferase dUTP nick end labeling assay) and expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) (immunoblotting). When compared with the control diet, apigenin lowered the number of high multiplicity ACF (HMACF >4 aberrant crypts/focus) by 57% ( P < 0.05), while naringenin lowered both the number of HMACF by 51% ( P < 0.05) and the proliferative index by 32% ( P < 0.05). Both apigenin and naringenin increased apoptosis of luminal surface colonocytes (78% and 97%, respectively; P < 0.05) when compared with the control diet. Hesperidin, nobiletin and the limonin glucoside/obacunone glucoside mixture did not affect these variables. The colonic mucosal protein levels of iNOS or COX-2 were not different among the six diet groups. The ability of dietary apigenin and naringenin to reduce HMACF, lower proliferation (naringenin only) and increase apoptosis may contribute toward colon cancer prevention. However, these effects were not due to mitigation of iNOS and COX-2 protein levels at the ACF stage of colon cancer.
Polyphenols enhance total oxidant-scavenging capacities of human blood by binding to red blood cells
Authors: Koren Erez , Kohen Ron , Ginsburg Isaac
Published: May 2010

The present study offers a new look at the role of erythrocytes and of erythrocytes–polyphenol complexes as potent ‘sinks’ for reactive oxygen species. We hereby show that human erythrocytes have the capacity not only to carry oxygen, but also to bind avidly to their surfaces a large variety of polyphenol antioxidants, which endows upon such complexes enhanced total oxidant-scavenging capacities (TOSC). This was proven by using confocal microscopy, 2,2-diphenyl-1-picrylhydrazyl radical, Folin-Ciocalteu's reagent, cyclic voltammetry and chemiluminescence techniques. The results presented suggest that the true TOSC of blood is the sum of intracellular antioxidants of red blood cells and other blood cells (mainly due to catalase), the polyphenols bound to their surfaces and the antioxidant agents present in plasma. Since erythrocytes can avidly bind and rapidly remove circulating polyphenols, the rule of the thumb to quantify antioxidants in health and disease processes exclusively in plasma as customary in clinical settings, does not represent the true TOSC of whole blood. We also postulate that circulating erythrocytes and possibly also other blood cells might be constantly coated by polyphenols from supplemented nutrients, which act as antioxidant depots and can thus act as protectors against the harmful consequences of oxidative stress. Further studies are needed to determine the faith of polyphenols in the circulation and their sequestration in the spleen.
Escherichia coli lipopolysaccharide impairs the calcium signaling pathway in mesangial cells: role of angiotensin II receptors
Authors: Maquigussa Edgar , Arnoni Carine P , Cristovam Priscila C , de Oliveira Andrea S , Higa Elisa M S , Boim Mirian A
Published: May 2010

Sepsis causes impaired vascular reactivity, hypotension and acute renal failure. The ability of the Escherichia coli endotoxin (lipopolysaccharide [LPS]) to impair agonist-induced contractility in mesangial cells, which contributes to LPS-induced renal dysfunction, was evaluated. Agonist-induced intracellular calcium ([Ca2+]i) mobilization was analyzed using angiotensin II (AngII). The effect of LPS on the levels of the renin–angiotensin system (RAS) components and the roles of vasodilatation-inducing molecules including AT2 receptor (AT2R) and nitric oxide (NO) in the cell reactivity were also evaluated. Confluent human mesangial cells (HMCs) were stimulated with LPS (0111-B4, 100 μg/mL). AngII-induced [Ca2+]i mobilization was measured by fluorometric analysis using Fura-2AM in the absence and presence of an AT2R antagonist (PD123319). The mRNA and protein levels for angiotensinogen, renin, angiotensin-converting enzyme, AT1R and AT2R were analyzed by realtime reverse transcriptase-polymerase chain reaction and Western blot, respectively. NO production was measured by the chemiluminescence method in the culture media after 24, 48 and 72 h of LPS incubation. After 24 h, LPS-stimulated HMCs displayed lower basal [Ca2+]i and an impaired response to AngII-induced rise in [Ca2+]i. LPS significantly increased AT2R levels, but did not cause significant alterations of RAS components. PD123319 restored both basal and AngII-induced [Ca2+]i peak, suggesting an involvement of AT2R in these responses. The expected increase in NO production was significant only after 72 h of LPS incubation and it was unaffected by PD123319. Results showed that LPS reduced the reactivity of HMCs to AngII and suggest that the vasodilatation induced by AT2R is a potential mediator of this response through a pathway independent of NO.
Effect of finger position on the median nerve area within the carpal tunnel: an ultrasound imaging study
Authors: Nadar Mohammed Shaban , Asbeutah Akram , Al-Obaidi Saud
Published: May 2010

Introduction

Finger flexion has been suggested to contribute to the aetiology of carpal tunnel syndrome (CTS). The purpose of this study was to measure the immediate effects of finger flexion on the cross-section area of the median nerve (MNA).

Methods

Thirty-one healthy volunteers participated in this experimental study. The MNA was measured at the level of hook of hamate using high-resolution ultrasonography during four different finger positions.

Results

A significant change in MNA during different finger positions was found ( F = 24.149, P < 0.001). The biggest difference in MNA was recorded between full finger extension and forceful finger flexion (mean difference = 1.29 mm2, P < 0.001). An inverse correlation between MNA and finger flexion was found (Pearson's r = −0.348, P = 0.002) where the MNA decreased as fingers moved toward flexion.

Conclusion

Finger flexion resulted in direct decrease in MNA, possibly contributing to median nerve compression.

Clinical implications

Hand therapists need to consider modifying their conservative treatment of CTS by limiting activities that require finger flexion and restricting the movement of the wrist and fingers to restrict finger flexion, thus limiting pressure increase on the median nerve.
Use of casting motion to mobilize stiffness to regain digital flexion following Dupuytren's fasciectomy
Authors: Midgley Robyn
Published: May 2010

Background

A priority in hand therapy following Dupuytren's fasciectomy is to regain and maintain extension of the proximal interphalangeal joint (PIPJ). Historically this has been achieved through the application of mechanical stress to the affected digit with splinting and exercise. However, loss of a normal pattern of digital flexion in both the operated and non-operated digits is a common complication.

Method

Four patients who underwent a Dupuytren's fasciectomy presented with persistent digital stiffness and a poor pattern of motion of the operated hand following traditional therapy. The casting motion to mobilize stiffness (CMMS) technique was therefore applied. The type of cast applied, duration of casting, duration of sessions and cast exercises are described.

Results

All patients regained digital flexion and an improved pattern of motion through the use of the CMMS technique without compromising PIPJ extension.

Conclusion

This small case series demonstrates the use of the CMMS technique to regain digital flexion and a normal pattern of motion when traditional treatment fails to achieve the desired outcomes following Dupuytren's fasciectomy. The use of the CMMS technique was beneficial in these patients and provides a means to regain digital flexion without the loss of PIPJ extension and is successful even when traditional therapy has failed.
Validity, responsiveness, intra- and inter-rater reliability of the weighted tape measure when measuring digital circumference
Authors: Jansen Victoria , Radbourne Liz , Fakis Apostolos , Bradley Mary , Burke Frank , Ellis Jo
Published: May 2010

Introduction

Circumferential measurements (CMs) are used by hand therapists to monitor digital oedema. There are a range of tape measures available commercially to measure digital circumference, but none of them have any data to support their reliability or validity.

Method

Observational measurement studies using a repeated measures design were undertaken to assess if a weighted tape measure, used with a protocol, could provide accurate and reliable CMs of swollen digits. In study 1, validity and responsiveness were determined by two raters measuring 16 steel bars of known circumferences and comparing the known measurements to those taken with the weighted tape. In study 2, intra- and inter-rater reliability was assessed by three raters measuring 12 prosthetic digits. In study 3, 27 swollen digits were measured by three raters to assess intra- and inter-rater reliability of the raters using the weighted tape in a clinical setting. The analysis of validity was based on the Bland–Altman plots. Reliability was based on the intraclass correlation coefficient (ICC).

Results

There was near-perfect agreement between the measurements taken by two assessors at the start and end of the trial and the actual circumference of the steel bars (all mean differences <1 mm). A minimum change in circumference of 2 mm could be detected with the same rater and a change of 3 mm could be detected with a different rater. On swollen digits the ICC values were significantly greater than 0.9 for both inter- and intrarater reliability. The lowest values for inter-rater reliability were ICC 0.983 (0.965–0.992) and for intrarater reliability were ICC 0.978 (0.954–0.990).

Discussion

The weighted tape measure has been shown to be valid and reliable in this study when used with a protocol for measuring swollen digits. It was easy to use and may become a clinically useful tool.
Paediatric cardiac telemedicine – transmitting echocardiography images in Wales
Authors: Lewis Delyth , Morris Sheryl , Uzun Orhan , Tranter Glynis
Published: May 2010

We started a telemedicine service for paediatric cardiology work in 2006, enabling transmission of echocardiography images via videoconferencing equipment to the specialist paediatric cardiac department at Cardiff for review and/or diagnosis. The system has been used regularly for a range of paediatric cardiology cases. Despite increasing the transmission bandwidth to 1 Mbit/s the transmission of live echocardiography was still considered to be suboptimum. Nonetheless, the implementation of a simple telemedicine service has demonstrated the feasibility of transmitting echocardiography images via videoconferencing equipment for diagnosis and second opinions.
Electromyographic audio biofeedback for telerehabilitation in hospital
Authors: Rogante Marco , Silvestri Salvatore , Grigioni Mauro , Zampolini Mauro
Published: May 2010

We investigated the use of surface electromyography with biofeedback for telerehabilitation. The aim was to investigate the therapist's point of view, and the general acceptability of the system for the patient and for the therapist. The system allowed a therapist to modulate the rehabilitation protocol remotely, by changing the configuration variables, e.g. threshold and duration. The case study focused on a patient with arm impairment following a stroke. After two weeks of daily usage, there were some differences between the patient and the therapist about the ease of use of the equipment. However, there was general agreement about the usefulness of the system, ease of the performed tasks, appropriateness of the performed tasks and overall opinion. Wireless technology would reduce the complexity of the tasks required of the patient.