[Multimodality navigation for liver resection of complicated alveolar echinococcosis].

Objective: To investigate the clinical application of multimodality navigation for liver resection in the treatment of complicated alveolar echinococcosis (AE). Methods: From October 2019 to February 2020, the clinical data and perioperative results of patients with AE treated by surgery in our department were retrospectively studied. Hepatic parenchyma disconnection plane and liver resection were navigated and performed with three-dimensional reconstruction and HITACHI real-time multi-image fusion interventional navigation system (RVS). Results: All of six patients were successful performed radical liver resection without mortality. The operation time was (301±106)min and the median blood loss was 200 ml. Two patients needed blood transfusion intraoperative (33.33%). The postoperative hospital stay was (10.8±2.8) day, and the cost of hospitalization was (82 584±995.61) yuan. Clavien-Dindo grade Ⅲ complication occurred in one patient. Conclusions: Multimodality navigation might provide precise intraoperative navigation of the surgical plane and effectively assist liver resection for the treatment of complicated AE.

Correlation between HSD17B4 expression in rat liver cancer tissues and inflammation or proliferation.

OBJECTIVE: Pathogenesis and progression of liver cancer are correlated with inflammatory response and estrogen level. 17ß-estradiol dehydrogenase IV (HSD17B4) is highly expressed in human liver cancer tissues. HSD17B4 participates in liver cancer cell proliferation via suppressing estradiol (E2) activity. This study generated a rat liver cancer model, on which the correlations between HSD17B4 and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), proliferating cell nucleus antigen (PCNA) expression were analyzed. MATERIALS AND METHODS: Male Sprague Dawley (SD) rats were randomly assigned into control and model group (N=30). Diethylnitrosamine was used to induce liver cancer in a rat model. HE staining was used to observe liver injury whilst ELISA was used to measure serum TNF-α and IL-6 levels. The level of serum E2 was quantified by radioimmunoassay. Serum liver function indexes were measured by automatic biochemical analyzer. Protein expressions of HSD17B4, p-Akt, p-ERK and PCNA were measured by Western blot. RESULTS: The inflammatory infiltration and necrosis of hepatocytes were shown in model group by HE staining, along with aggravated liver indexes. Significantly high phosphorylation level of Akt and ERK, along with the increase of HSD17B3 and PCNA expressions, was found in model group (p<0.05 compared to control group). Serum E2 level was statistically decreased, whilst TNF-α and IL-6 were up-regulated (p<0.05). HSD17B4 was positively correlated with TNF-α, IL-6 and PCNA expressions (r=0.68, 0.62 and 0.56, p<0.05). CONCLUSIONS: HSD17B4 is over-expressed in rat liver cancer tissues. Its expression was positively correlated with TNF-α, IL-6 and PCNA levels, and probably participates in liver cancer cell proliferation via ERK and Akt signal pathway.

Bone biomechanical properties in EP4 knockout mice.

Among the four prostaglandin E receptor subtypes, EP(4) has been implicated as an important regulator of both bone formation and bone resorption; however, the integrated activities of this receptor on bone biomechanical properties have not been examined previously. This study compared the bone biomechanical properties of EP(4) knockout (KO) transgenic mice to strain-matched wild-type (WT) controls. We examined two groups of adult female mice: WT (n = 12) and EP(4) KO (n = 12). Femurs were tested in three-point bending and the lumbar-4 (L4) vertebral body by compression. Distal femur and vertebral body trabecular bone architecture were quantified using micro-computed tomography. Biomechanical structural parameters (ultimate/yield load, stiffness) were measured and apparent material parameters (ultimate/yield stress, modulus) calculated. Body weights and bone sizes were not different between EP(4) KO and WT mice (P > 0.05, Student's t-test). EP(4) KO mice exhibited reduced structural (ultimate/yield load) and apparent material (ultimate/yield stress) strength in the femoral shaft and vertebral body compared to WT (P < 0.05). Vertebral body stiffness and femoral neck ultimate load (structural strength) were marginally lower in EP(4) KO than that in WT mice (P < 0.1). In addition, EP(4) KO mice have smaller distal femur and vertebral bone volume to total volume (BV/TV) trabecular thickness than WT mice (P < 0.05). These results suggest that the prostaglandin receptor EP(4) has an important role in determining biomechanical competence in the mouse skeleton. Despite similar bone size, the absence of an EP(4) receptor may have removed a necessary link for bone adaptation pathways, which resulted in relatively weaker bone properties.

Surface-specific effects of a PPARgamma agonist, darglitazone, on bone in mice.

It has been hypothesized that activation of peroxisome-proliferator-activated receptor-gamma (PPARgamma) by thiazolidinedione drugs can increase adipogenesis at the expense of osteogenesis, leading to bone loss. However, the reported skeletal effects of these compounds are varied and their effects on cortical bone are unknown. In this study, we examined the changes in both cancellous and cortical bone of 6-month-old male mice treated with darglitazone, a potent and selective PPARgamma agonist, at 10 mg/kg/day by dosing the compound in a food mixture for 2 or 8 weeks. At 2 weeks, we observed significantly increased marrow adipose tissue area, decreased trabecular bone density of distal femur, and decreased surface referent bone formation rate of lumbar vertebrae in the mice treated with darglitazone compared with controls. At 8 weeks, lower cancellous bone mass was seen at both distal femurs and lumbar vertebrae of the mice treated with darglitazone. In addition, mineralizing surface was significantly lower, whereas osteoclast surface and number were significantly higher in the lumbar vertebrae of darglitazone-treated mice. At the femoral diaphysis, darglitazone treatment caused bone loss on the endocortical surface. Interestingly, periosteal mineral apposition rate and surface referent bone formation rate were significantly increased in darglitazone-treated mice. In bone marrow cell cultures, darglitazone suppressed alkaline phosphatase activity, osteoblastic gene expression, and mineralized nodule formation while increasing adipogenic gene expression and lipid accumulation. In summary, darglitazone enhanced adipogenesis and caused cancellous bone loss by increasing bone resorption and decreasing bone formation in mice. In addition, darglitazone induced cortical bone loss on the endocortical surface but increased bone formation on the periosteal surface. These data suggest that PPARgamma plays a role in regulating bone resorption and formation and reveal surface-specific effects of a PPARgamma agonist on bone.

Fully automated large-scale assessment of visceral and subcutaneous abdominal adipose tissue by magnetic resonance imaging.

OBJECTIVE: To describe and evaluate a fully automated method for characterizing abdominal adipose tissue from magnetic resonance (MR) transverse body scans. METHODS: Four MR pulse sequences were applied: SE, FLAIR, STIR, and FRFSE. On 39 subjects, each abdomen was traversed by 15 contiguous transaxial images. The total abdominal adipose tissue (TAAT) was calculated from thresholds obtained by slice histogram analysis. The same thresholds were also used in the manual volume calculation of TAAT, subcutaneous abdominal adipose tissue (SAAT) and visceral abdominal adipose tissue (VAAT). Image segmentation methods, including edge detection, mathematical morphology, and knowledge-based curve fitting, were used to automatically separate SAAT from VAAT in various 'nonstandard' cases such as those with heterogeneous magnetic fields and movement artefacts. RESULTS: The percentage root mean squared errors of the method for SAAT and VAAT ranged from 1.0 to 2.7% for the four sequences. It took approximately 7 and 15 min to complete the 15-slice volume estimation of the three adipose tissue classes using automated and manual methods, respectively. CONCLUSION: The results demonstrate that the proposed method is robust and accurate. Although the separation of SAAT and VAAT is not always perfect, this method could be especially helpful in dealing with large amounts of data such as in epidemiological studies.

Osteopenia and impaired fracture healing in aged EP4 receptor knockout mice.

The EP4 receptor, one of the subtypes of the prostaglandin E2 (PGE2) receptor, plays a critical role in the anabolic effects of PGE2 on bone. However, its role in the maintenance of bone mass in aged animals and its role in fracture healing is not well known. Our studies addressed these issues by characterizing the skeletal phenotype of aged, EP4 receptor knockout (KO) mice, and by comparing fracture healing in aged KO mice versus wild type (WT) mice. There was no significant difference in body weight and femoral length between KO and WT mice at 15 to 16 months of age. Lower bone mass was seen radiographically in both axial and long bones of KO mice relative to WT mice. Micro-CT images of the distal femurs showed thinner cortices, fewer trabeculae, and a deteriorated trabecular network in KO mice. Total bone content, trabecular content, and cortical content, as assessed by pQCT in the distal femur, were lower in KO mice than WT controls. Histomorphometric measurements showed that trabecular bone volume and bone formation rate were significantly decreased whereas osteoclast number on trabecular surface and eroded surface on endocortical surface were significantly increased in KO mice. These data indicated that deleting the EP4 receptor resulted in an imbalance in bone resorption over formation, leading to a negative bone balance. The lower bone formation rate in EP4 KO mice was primarily due to decreased mineralizing surface, suggesting that the defect in overall bone formation was mainly due to the defect in osteoblastogenesis. Fracture healing was examined in KO and WT mice subjected to a transverse femoral fracture. Callus formation was significantly delayed as evidenced both radiographically and histologically in the fractured femurs of KO mice compared with those of WT mice. KO mice had significant decreases in total callus area, cartilaginous callus area, and bony callus area 2 weeks after fracture. By 4 weeks, complete bony bridging was seen in WT mice but not in KO mice. These data demonstrate that the absence of the EP4 receptor decreases bone mass and impairs fracture healing in aged male mice. Our findings indicate that the EP4 receptor is a positive regulator in the maintenance of bone mass and fracture healing.

Efficacy of a growth hormone-releasing peptide mimetic in cardiac ischemia/reperfusion injury.

The cardioprotective efficacy of the pyrazolinone-piperidine dipeptide growth hormone secretagogue (GHS) CP-424,391 was studied in an in vivo rabbit model of ischemia and reperfusion. CP-424,391 was administered at 25 mg/kg p.o. x 7 days. Ischemia was induced by left coronary artery occlusion for 30 min, after which the heart was reperfused for 2 h. At the end of reperfusion, animals were euthanized and the infarct size was determined. The area at risk of infarct was not different between the control (45.8+/-3.7%, n=6) and CP-424,391-treated groups (36.9+/-4.3%, n=11). The infarct size of the control animals was 49.5+/-7.1% and was significantly (P<0.05) lower in the CP-424,391-treated group (infarct size=17.3+/-3.0). There was a trend, albeit not significant, for the left ventricular function to recover to a greater extent in CP-424,391-treated rabbits. Thus, the treatment of rabbits for 7 days with CP-424,391 was cardioprotective against ischemia/reperfusion injury.

Flexural behaviour of post-cured composites at oral-simulating temperatures.

Post-curing treatments have been known to improve the mechanical stability of visible light-cured composites. After individual post-curing treatment, the flexural strength (FS) of four commercial direct/indirect placement composite materials which differ greatly in composition [oligocarbonate dimethacrylate (OCDMA)-based Conquest C & B (CQT), Bisphenol-A glycidyl dimethacrylate (BisGMA)-based Charisma, urethane dimethacrylate (UDMA)-based Concept (CCT), and BisGMA/UDMA-based Dentacolor] was evaluated under water in the temperature range of 12-50 degrees C. A control series was tested in air at room temperature (25 +/- 1 degrees C). Data were analysed using ANOVA and Duncan's test. Flexural strengths overall decreased (20-40%, P < 0.01) with increasing temperatures except with Conquest C & B. Surprisingly, higher FS values were found in wet conditions than in dry conditions at 25 degrees C. UDMA-based materials much more easily undergo softening in water and by temperature change than do BisGMA- or OCDMA-based materials. Post-cured composites can be significantly affected by exposure to oral environments. Different composition determines the degree of influence.

Mismatch negativity and personality traits in chronic primary insomniacs.

The thalamo-(fronto)cortical circuit is involved in sleep regulation, and its dysfunction might contribute to the pathophysiology of chronic primary insomnia. To obtain more evidence of the involvement of the circuit, we studied 23 patients with chronic primary insomnia and 28 healthy volunteers via the assessment of mismatch negativity (MMN) elicited by tone intensity deviance, and of personality traits measured by Zuckerman's Sensation Seeking Scales, and Zuckerman-Kuhlman's Personality Questionnaire. In insomniacs, MMN amplitude at Fz was significantly larger; Depression, which was measured by Plutchik-van Praag's Depression Inventory, and Neuroticism-anxiety and Impulsivity scores were higher, while the Thrill and adventure seeking score was lower; MMN amplitude was positively correlated with Depression and with Impulsivity. In healthy subjects, MMN amplitude at Fz was positively correlated with Neuroticism-anxiety, but negatively with Experience seeking. The larger MMN and distinct personality traits suggest a hyperactivity in the thalamo-(fronto)cortical neuronal circuit in insomniacs, which is probably the result of weak thalamic gating mechanisms, or an imbalance of several neurotransmitter systems.

Factors influencing the dynamic behaviour of human teeth.

Modal analysis is carried out to test the natural frequencies of certain human teeth, including central incisors (CIs), canines (CAs), first premolars (FPs) and first molars (FMs). A total number of 1007 teeth are tested, taking into account tooth type, oral location, age and gender, to analyse the effects of the above-mentioned factors on the natural frequency of the sample teeth. The results reveal that no significant difference in the natural frequency is noted among teeth in the four different intra-oral quadrants. Nevertheless, tooth type and age elicit an effect upon the value of the natural frequency of teeth. On the other hand, the mean value for the natural frequency of CIs (1.27 +/- 0.15 kHz), CAs (1.30 +/- 0.15 kHz), FPs (1.27 +/- 0.15 kHz) and FMs (1.16 +/- 0.12 kHz) for males are significantly lower (p < 0.01) than the analogous figure for females (1.41 +/- 0.21 kHz for CIs, 1.40 +/- 0.18 kHz for CAs, 1.37 +/- 0.20 kHz for FPs, and 1.25 +/- 0.16 kHz for FMs). Moreover, the natural frequency of teeth in male subjects varies with age (p < 0.05). The highest mean frequency of CIs, CAs and FPs for the male subjects is found for the group aged between 40 and 49 years. On the other hand, the natural frequency for the similar set of teeth for the female subjects is shown to be in no way associated with age.

Preclinical pharmacology of CP-424,391, an orally active pyrazolinone-piperidine [correction of pyrazolidinone-piperidine] growth hormone secretagogue.

Growth hormone secretagogues (GHSs) represent attractive therapeutic alternatives to recombinant growth hormone (GH), given their ability to amplify pulsatile hormone secretion in a relatively physiologic manner. CP-424,391 (391) is a novel, orally active pyrazolinone-piperidine [corrected] GHS. In rat pituitary cell cultures, 391 stimulated GH release with an EC50 = 3 nM. The addition of 391 to rat pituitary cells activated intracellular calcium signaling but did not elevate intracellular cyclic adenosine monophosphate (cAMP). 391 also modulated the effects of GH-releasing hormone and somatostatin on pituitary cell GH-release and intracellular signaling. In nonpituitary cell lines, the ability of 391 to stimulate intracellular signaling was dependent on the expression of recombinant human GHS receptor. Acute administration of 391 to anesthetized rats or to conscious dogs induced pulsatile release of G H in a dose-dependent manner. Plasma insulin-like growth factor-I (IGF-I) was elevated progressively over a 5-d course of daily oral dosing in dogs. Chronic oral administration of 391 augmented body weight gain in rats and dogs. Thus, the peptidomimetic GHS 391 has potential utility for the treatment of clinical conditions that could benefit from systemic augmentation of GH and IGF-I levels.

Improving biomaterial properties of collagen films by chemical modification.

Films of bovine collagen were chemically modified with the goal of improving their biomaterial properties. The modified films were investigated with respect to their affinity to fibroblast and endothelial cells, as well as their antibacterial properties tested by adhesion of Staphylococcus aureus. Modifications that only change the net charge of collagen, such as acetylation, succinylation, and treatment with glutaraldehyde (all increase the negative charge), and amination with ethylenediamine (EDA), N,N-dimethyl-EDA (DMEDA), or butylamine (all increase the positive charge), did not dramatically alter the mammalian cell attachment to the film. In contrast, derivatization of collagen using methoxypoly(ethylene glycol) (PEG) diminished the attachment of fibroblasts by 98 +/- 1% and of endothelial cells by more than 99% compared to unmodified collagen. Moreover, the rate of growth of fibroblasts dropped by 97 +/- 1% and that of endothelial cells by 88 +/- 3% as a result of PEGylation of collagen. Adhesion of S. aureus cells also plummeted by 93 +/- 2% as a result of this PEGylation. With these antifouling properties, PEG-collagen may be a promising coating material for coronary stents. Subsequent derivatization of PEG-collagen with EDA or DMEDA abolished its mammalian cell-repelling ability, whereas bacterial cell repulsion was partially retained: for example, DMEDA-modified PEG-collagen exhibits up to a 5-fold lower bacterial adhesion than collagen. It is worth noting that a material that allows mammalian cell attachment but reduces bacterial adhesion could be useful as an implant or coating.

Treatment with a growth hormone secretagogue in a model of developing heart failure: effects on ventricular and myocyte function.

BACKGROUND: Exogenous administration of growth hormone (GH) and subsequently increased production of insulin-like growth factor-1 can influence left ventricular (LV) myocardial growth and geometry in the setting of congestive heart failure (CHF). This study determined the effects of an orally active GH secretagogue (GHS) treatment that causes a release of endogenous GH on LV function and myocyte contractility in a model of developing CHF. METHODS AND RESULTS: Pigs were randomly assigned to the following treatment groups: (1) chronic rapid pacing at 240 bpm for 3 weeks (n=11); (2) chronic rapid pacing and GHS (CP-424,391 at 10 mg x kg(-1) x d(-1), n=9); and (3) sham controls (n=8). In the untreated pacing CHF group, LV fractional shortening was reduced (21+/-2% versus 47+/-2%) and peak wall stress increased (364+/-21 versus 141+/-5 g/cm(2)) from normal control values (P:<0.05). In the GHS group, LV fractional shortening was higher (29+/-2%) and LV peak wall stress lower (187+/-126 g/cm(2)) than untreated CHF values (P:<0.05). With GHS treatment, the ratio of LV mass to body weight increased by 44% from untreated values. Steady-state myocyte velocity of shortening was reduced with pacing CHF compared with controls (38+/-1 versus 78+/-1 microm/s, P:<0.05) and was increased from pacing CHF values with GHS treatment (55+/-7 microm/s, P:<0.05). CONCLUSIONS: The improved LV pump function that occurred with GHS treatment in this model of CHF was most likely a result of favorable effects on LV myocardial remodeling and contractile processes. On the basis of these results, further studies are warranted to determine the potential role of GH secretagogues in the treatment of CHF.

Assessing the implant/bone interface by using natural frequency analysis.

OBJECTIVE: A number of techniques have been proposed for detecting the stability of dental implants. However, the clinical applicability of those methods is still limited. The purpose of this study was to evaluate a new innovative, noninvasive, minimum-contact method for the stability assessment of dental implants. STUDY DESIGN: Natural frequency is a physical property of a structure, which is strongly related to its boundary conditions. In this study, a modal testing technique was carried out to measure the natural frequency of dental implants. The implants were fixed by a metal clamp stand and were excited to vibrate by an impulse hammer. A noncontact piezoelectric microphone then acoustically acquired the vibration responses of the implants. Natural frequencies of the tested implants were recorded under various clamping forces and clamping levels. RESULTS: Natural frequencies of the tested implants were concentrated from 8 to 19 kHz under different boundary conditions. On the other hand, the natural frequency values decreased when boundary levels and boundary force were reduced. Linear relationships (P <.005) were found between response frequencies and the degree of implant stability. CONCLUSIONS: Our results show that the boundary status of an implant can be monitored by detecting its natural frequency. A noncontact transducer used in this study can also serve as a useful tool for future clinical investigations.

Finite element analysis of brain contusion: an indirect impact study.

The mechanism of brain contusion has been investigated using a series of three-dimensional (3D) finite element analyses. A head injury model was used to simulate forward and backward rotation around the upper cervical vertebra. Intracranial pressure and shear stress responses were calculated and compared. The results obtained with this model support the predictions of cavitation theory that a pressure gradient develops in the brain during indirect impact. Contrecoup pressure-time histories in the parasagittal plane demonstrated that an indirect impact induced a smaller intracranial pressure (-53.7 kPa for backward rotation, and -65.5 kPa for forward rotation) than that caused by a direct impact. In addition, negative pressures induced by indirect impact to the head were not high enough to form cavitation bubbles, which can damage the brain tissue. Simulations predicted that a decrease in skull deformation had a large effect in reducing the intracranial pressure. However, the areas of high shear stress concentration were consistent with those of clinical observations. The findings of this study suggest that shear strain theory appears to better account for the clinical findings in head injury when the head is subjected to an indirect impact.

Laser-induced acoustic emissions in experimental dental composites.

A laser thermoacoustic technique was innovated to evaluate laser-induced acoustic emissions (AEs) in experimental dental composites aged with 75% ethanol solution. Experimental composite systems of 75/25 BisGMA/TEGDMA resin filled with 0, 12.6, 30.0, and 56.5 vol% of 8-microm silanized and unsilanized BaSiO6 were analyzed. The sample size was 4.65 mm (diameter) x 0.5 mm (thick). Aging effects of immersing in 75% ethanol for up to 14 h on AEs were then evaluated. A continuous-wave CO2 laser was used to heat the samples. Acoustic emissions were collected as a function of filler fraction, laser power, silanization, and immersion time. Onset of burst-pattern acoustic signals characteristic of fracturing occurred at different laser powers for different tested groups. Acoustic emissions generally increased with laser power, in which lower laser powers produced low-amplitude (45-50 dB) signals; the amplitude distribution (50-85 dB) became more extensive as laser powers increased. After immersion, the lower laser powers could produce the same phenomenon. The higher the filler fraction, the fewer AEs generated. A large percentage AE reduction due to silanization was noted as a function of filler fraction. Unsilanized specimens showed more thermal damages than did silanized ones.

Effect of temperature and flux concentration on soldering of base metal.

The present study used the acoustic emission (AE) technique to evaluate interactions among soldering temperature, flux treatment, and the resultant ultimate tensile strength (UTS). Scanning electron microscopy (SEM) was used to examine fracture surfaces of the solder joints. Specimens were cast from removable partial denture alloy and then placed in a jig with a gap distance of 1.0 mm. A high-frequency soldering machine with an optical pyrometer was used for soldering at 1150 degrees C and 1200 degrees C, respectively. The flux concentrations were 67% and 75%. The soldered specimens were subjected to tensile test at a crosshead speed of 0.05 mm/min. During testing, acoustic emissions in the frequency range of 100--1200 kHz were collected, filtered, recorded, and processed by a sensing device. The results were analysed by ANOVA and Tukey LSD test. UTS at different temperatures showed no significant difference according to either mechanical or acoustic results. But in the 1200 degrees C group, the UTSs and AE counts showed significant differences (P<0.05) at both flux concentrations. SEM showed that the 1200C group had better dendritic crystal structure than did the 1150 degrees C group. In the 1200 degrees C group specimens with 67% flux had fewer flux inclusion bodies and dendritic crystals than did specimens with 75% flux. The 75% flux subgroup produced high-amplitude (60--70 dB) acoustic signals within the elastic deformation zone, while the 67% flux subgroup produced similar signals within the plastic deformation zone, either beyond the 0.2% yield point or before fracture.

Effects of growth hormone supplementation on left ventricular morphology and myocyte function with the development of congestive heart failure.

BACKGROUND: Release of growth hormone (GH), putatively through alterations in insulin growth factor-1 (IGF-1) levels, has been implicated to influence left ventricular (LV) myocardial structure and function. The objective of this study was to determine contributory mechanisms by which GH supplementation may influence LV function with the development of congestive heart failure (CHF). METHODS AND RESULTS: Pigs were assigned to the following groups: (1) chronic pacing at 240 bpm for 3 weeks (n = 10), (2) chronic pacing and GH supplementation (200 microg x kg(-1) x d(-1), n = 10), and (3) controls (n = 8). GH treatment increased IGF-1 plasma levels by nearly 2.5-fold throughout the pacing protocol. In the untreated pacing CHF group, LV fractional shortening was reduced and peak wall stress increased. In the pacing CHF and GH groups, LV fractional shortening was higher and LV wall stress lower than untreated CHF values. Steady-state myocyte velocity of shortening was reduced with pacing CHF and was unchanged from CHF values with GH treatment. In the presence of 25 nmol/L isoproterenol, the change in myocyte shortening velocity was reduced in the untreated CHF group and increased in the GH-treated group. LV sarcoplasmic reticulum Ca(2+)-ATPase abundance was reduced with pacing CHF but was normalized with GH treatment. CONCLUSIONS: Short-term GH supplementation improved LV pump function in pacing CHF as a result of favorable effects on LV remodeling and contractile processes. Thus, GH supplementation may serve as a novel therapeutic modality in developing CHF.

Cloning, structural characterization, and chromosomal localization of the gene encoding the human prostaglandin E(2) receptor EP2 subtype.

Northern blot analysis of human placental RNA using a probe to the 5' end of the human prostaglandin E(2) (PGE(2)) EP2 receptor subtype coding region revealed the existence of a high abundance, low molecular weight transcript. To investigate the origin of this transcript, and its possible relationship to the human EP2 mRNA, we have cloned and characterized the gene encoding the human PGE(2) EP2 receptor subtype, identified transcriptional initiation and termination sites in two tissues (spleen and thymus), and determined its chromosomal localization. The human EP2 gene consists of two exons separated by a large intron, utilizes a common initiation site in both spleen and thymus at 1113 bp upstream of the translation initiation site, and has 3' transcript termini at 1140 bp and 1149 bp downstream of the translation stop site in spleen and thymus respectively. Southern and fluorescence in situ hybridization analysis demonstrated the human EP2 gene to be a single copy gene located in band 22 of the long arm of chromosome 14 (14q22). Though our initial interest in this gene was to investigate potential differential splicing of the human EP2 gene in placenta, this work demonstrates that the atypical transcript observed in placenta probably arises from a distinct, yet related, gene. Knowledge of the sequence, structure, and transcription events associated with the human EP2 gene will enable a broader understanding of its regulation and potential role in normal physiology and disease.

Molecular cloning and characterization of the canine prostaglandin E receptor EP2 subtype.

Prostaglandin E2 (PGE2) binds to four G-protein coupled cell surface receptors (EP1-EP4) and has been implicated as a local mediator of bone anabolism via a cyclic AMP mediated pathway following activation of the EP2 and/or EP4 receptor subtype. A canine kidney cDNA library was screened using a human EP2 probe, and a clone with an open reading frame of 1083 bp, potentially encoding a protein of 361 amino acids, was characterized. This open reading frame has 89% identity to the human EP2 cDNA at the nucleotide level and 87% identity at the predicted protein level. Scatchard analysis of a CHO cell line stably transfected with canine EP2 yielded a dissociation constant of 22 nM for PGE2. Competition binding studies, using 3H-PGE2 as ligand, demonstrated specific displacement by PGE2, Prostaglandin E1, Prostaglandin A3, and butaprost (an EP2 selective ligand), but not by ligands with selectivity for the related DP, FP, IP, or TP receptors. Specific ligand binding also resulted in increased levels of cAMP in EP2 transfected cells with no evidence of short-term, ligand-induced desensitization. Northern blot analysis revealed two transcripts of 3300 and 2400 bp in canine lung, and reverse-transcription polymerase chain reaction showed expression in all tissues examined. Southern blot analysis suggests the presence of a single-copy gene for EP2 in the dog.