Biomedical subjects

S J Cho

Publications and source records attributed to S J Cho.

At least 37 records · Page 2Linked to original sources

PubMed2000

Usefulness of triphasic perfusion computed tomography for intravenous thrombolysis with tissue-type plasminogen activator in acute ischemic stroke.

BACKGROUND: Intravenous thrombolysis for acute ischemic stroke has been investigated in several clinical trials without enough information on collateral blood flow and perfusion deficit in the ischemic areas. The therapeutic time window varies from patient to patient depending on these factors. Triphasic perfusion computed tomography (TPCT) can provide this information as reliably as conventional angiography. OBJECTIVE: To assess the safety and efficacy of thrombolysis within 3 or 7 hours of stroke onset according to the extent of perfusion deficit on TPCT. METHODS: In 46 patients with acute middle cerebral artery (MCA) territory stroke, TPCT was performed with power injector-controlled, intravenous administration of contrast media after taking precontrast CT scans. Sequential scans of early, middle, and late phases were performed. The entire procedure took 5 minutes. Depending on collateral blood flow, the perfusion deficit on TPCT was graded as "severe perfusion deficit" or "moderate perfusion deficit." Twenty-nine patients were excluded based on clinical, laboratory, and TPCT findings. Seventeen patients were treated with an intravenous recombinant tissue-type plasminogen activator, 0.9 mg/kg. The 17 treated patients were divided into 2 groups: group 1 with small severe perfusion deficit (</=33% of the presumed MCA territory) and group 2 with medium-sized severe perfusion deficit (>33% but </=50% of the presumed MCA territory). The 13 patients in group 1 were treated within 7 hours of onset and the 4 patients in group 2 were treated within 3 hours. RESULTS: Initial mean National Institutes of Health Stroke Scale score was 12.1 (range, 6.0-20.0) in group 1 and 19.0 (range, 18.0-21. 0) in group 2. The initial score correlated better with the total extent of moderate perfusion deficit and severe perfusion deficit than that of severe perfusion deficit alone. Mean time lapse to thrombolysis was 4.2 hours (range, 1.5-7.0 hours) in group 1 and 2.2 hours (range, 1.9-2.5 hours) in group 2. Eight patients (47%), 7 from group 1 and 1 from group 2, improved by 4 points or more from baseline Stroke Scale score within 24 hours of thrombolysis. Patients with moderate perfusion deficit of 50% or more of MCA territory (n = 4) had a better chance of early improvement than did those (n = 13) with moderate perfusion deficit of less than 50% (4 of 4 vs 4 of 13). No fatal hemorrhage occurred. Only 1 patient (6%) had symptomatic small basal ganglia hemorrhage after thrombolysis. CONCLUSIONS: Thrombolysis can be safely performed within 3 or 7 hours of stroke onset according to the extent of severe perfusion deficit on TPCT. A larger extent of moderate perfusion deficit on TPCT may predict early improvement after thrombolysis.

Adult↗

PubMed2000

Triphasic perfusion computed tomography in acute middle cerebral artery stroke: a correlation with angiographic findings.

OBJECTIVE: To evaluate the usefulness of triphasic perfusion computed tomography (TPCT) in diagnosing middle cerebral artery (MCA) occlusion and in assessing the perfusion deficit and collateral circulation in patients with acute ischemic stroke. BACKGROUND: Conventional angiography is the criterion standard for the diagnosis of MCA occlusion and for the assessment of perfusion deficit and collateral blood supply. The risk of hemorrhagic transformation after recanalization of occluded arteries by thrombolytic therapy is considered high when pretherapeutic residual flow is markedly reduced. PATIENTS AND METHODS: In 8 patients within 3 hours of onset of acute MCA stroke, precontrast computed tomographic scans were taken, and then TPCT was performed after power-injector controlled intravenous administration of contrast media. Sequential images of early, middle, and late phases were obtained. The whole procedure took 5 minutes. Perfusion deficit on TPCT was graded as "severe" or "moderate," depending on the state of collateral flow. Digital subtraction angiography (DSA) was performed in all patients within 6 hours of acute stroke. Direct intra-arterial urokinase infusion was begun immediately after the angiographic superselection of the MCA occlusion site in 6 of the 8 patients within 7 hours of onset (range, 4.3-6.2 hours). RESULTS: The DSA findings showed occlusion of the MCA stem (n = 1) and at the bifurcation (n = 4). The sites of proximal MCA occlusion could be identified on the early and middle images of TPCT in all 5 patients. On DSA findings, all 8 patients had a zone of perfusion deficit with markedly slow leptomeningeal collaterals and a zone of perfusion deficit with no collaterals. The zone of severe perfusion deficit on TPCT corresponded to the zone of perfusion deficit with no or few collaterals on angiography, and the zone of moderate perfusion deficit on TPCT corresponded to that of perfusion deficit with markedly slow leptomeningeal collaterals. Early parenchymal hypoattenuation on precontrast computed tomography was confined to the zone of severe perfusion deficit on TPCT. The initial National Institutes of Health Stroke Scale score correlated better with the total extent of severe perfusion deficit and moderate perfusion deficit on TPCT than that of severe perfusion deficit alone. After direct intra-arterial thrombolysis within 7 hours of onset, symptomatic hemorrhagic transformation did not develop in 4 patients with small severe perfusion deficit (33% or less of the presumed MCA territory). However, the remaining 2 patients with large severe perfusion deficit (more than 50% of the presumed MCA territory) deteriorated to death with hemorrhagic transformation. CONCLUSIONS: Triphasic perfusion computed tomography is useful for diagnosing proximal MCA occlusion and assessing perfusion deficit and collateral circulation as reliably as DSA. The zone of severe perfusion deficit on TPCT may be presumed to be the ischemic core, and that of moderate perfusion deficit, the penumbra zone. Triphasic perfusion computed tomography may be used as a rapid and noninvasive tool to make thrombolysis safer.

Acute Disease↗

PubMed2000

Crystallization and preliminary X-ray diffraction analysis of Saccharomyces cerevisiae Ygr203p, a homologue of Acr2 arsenate reductase.

Ygr203p, a 148-residue protein encoded by the ygr203w gene of Saccharomyces cerevisiae, is a homologue of the yeast Acr2 arsenate reductase encoded by the acr2 (or ypr200c) gene. It also shows significant sequence similarity to the human cell-cycle control Cdc25 phosphatase family. It has been overexpressed in soluble form in Escherichia coli with a His(6) tag at its C-terminus. The recombinant protein has been crystallized at 296 K using sodium chloride as precipitant. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 40.48, b = 50.95, c = 91.95 A. The asymmetric unit contains a monomer, giving a crystal volume per protein mass (V(m)) of 2.61 A(3) Da(-1) and a solvent content of 53.8%. The crystals diffract to better than 1.9 A resolution with Cu Kalpha X-rays. They are therefore suitable for high-resolution structure determination.

Adenosine Triphosphatases↗

PubMed2000

Crystallization and preliminary X-ray crystallographic analysis of Escherichia coli CyaY, a structural homologue of human frataxin.

CyaY is a 106-residue protein from Escherichia coli. It shows amino-acid sequence similarity to human frataxin and a frataxin homologue in Saccharomyces cerevisiae, Yfh1p. The former is associated with the disease Friedreich ataxia and the latter plays a key role in iron homeostasis in mitochondria. CyaY has been overexpressed in soluble form in E. coli. The recombinant protein with a His(6) tag at its C-terminus has been crystallized at 296 K using polyethylene glycol (PEG) 4000 as a precipitant. Native diffraction data have been collected to 1.8 A using Cu Kalpha X-rays. The crystals belong to the trigonal space group P3(1)21 (or P3(2)21), with unit-cell parameters a = b = 44.66, c = 99.87 A, alpha = beta = 90.0, gamma = 120.0 degrees. The asymmetric unit contains one molecule of recombinant CyaY, with a corresponding V(m) of 2.13 A(3) Da(-1) and solvent content of 42.3%.

Bacterial Proteins↗

PubMed2000

Lactate dehydrogenase from the hyperthermophilic archaeon Methanococcus jannaschii: overexpression, crystallization and preliminary X-ray analysis.

L(+)-Lactate dehydrogenase (LDH) is a key enzyme in anaerobic metabolism which converts pyruvate to lactate. LDH from the hyperthermophilic archaebacterium Methanococcus jannaschii has been overexpressed in Escherichia coli and crystallized in two crystal forms at 297 K using 2-methyl-2,4-pentanediol as precipitant. Type I crystals grew rapidly and diffracted to at least 2.8 A Bragg spacing upon exposure to Cu Kalpha X-rays. X-ray diffraction data to 2.9 A have been collected from a native crystal. The type I crystal is tetragonal, belonging to the space group P4(2)2(1)2, with unit-cell parameters a = b = 99.74, c = 170.00 A. The asymmetric unit contains two LDH subunits, with a corresponding crystal volume per protein mass (V(m)) of 3.05 A(3) Da(-1) and a solvent content of 59.7%. Type II crystals, which grew more slowly, diffracted to at least 1.8 A Bragg spacing upon exposure to Cu Kalpha X-rays. X-ray diffraction data to 1.9 A have been collected from a native crystal. The type II crystal is orthorhombic, belonging to the space group P2(1)2(1)2, with unit-cell parameters a = 47.65, b = 125.10, c = 58.08 A. The asymmetric unit contains a single LDH subunit, with a corresponding crystal volume per protein mass (V(m)) of 2.50 A(3) Da(-1) and a solvent content of 50.8%. Therefore, the type II crystal is more suitable for high-resolution structure determination than the type I crystal.

Cloning, Molecular↗

PubMed2000

Cloning and sequencing of pel gene responsible for CMCase activity from Erwinia chrysanthemi PY35.

The phytopathogenic bacterium Erwinia chrysanthemi secretes multiple isozymes of plant cell wall disrupting enzymes such as pectate lyase and endoglucanase. We cloned genomic DNA from Erwinia chrysanthemi PY35. One of the E. coli XL1-Blue clones contained a 5.1-kb BamHI fragment and hydrolyzed carboxymethyl cellulose and polygalacturonic acid. By subsequent subcloning, we obtained a 2.9-kb fragment (pPY100) that contained the pel gene responsible for CMCase and pectate lyase activities. The pel gene had an open reading frame (ORF) of 1,278 bp encoding 425 amino acids with a signal peptide of 25 amino acids. Since the deduced amino acid sequence of this protein was very similar to that of PelL of E. chrysanthemi EC16, we concluded that it belonged to the pectate lyase family EC 4.2.2.2, and we designated it PelL1. Sequencing showed that the PeIL1 protein contains 400 amino acids and has a calculated pI of 7.15 and a molecular mass of 42,925 Da. The molecular mass of PelL1 protein expressed in E. coli XL1-Blue, as analyzed by SDS-PAGE, appeared to be 43 kDa. The optimum pH for its enzymatic activity was 9, and the optimum temperature was about 40 decreased C.

Amino Acid Sequence↗

PubMed2000

Crystallization and preliminary X-ray crystallographic analysis of human nucleoside diphosphate kinase A.

Human nucleoside diphosphate kinase A catalyzes phosphoryl transfer and acts as a suppressor of metastasis. It has been crystallized using 2-methyl-2,4-pentanediol as a precipitant at 288 K. The crystal is monoclinic, belonging to the space group P2(1), with unit-cell parameters a = 74.21, b = 78.11, c = 82.29 A, beta = 101. 33 degrees. The asymmetric unit contains a homohexamer, with a corresponding crystal volume per protein mass (V(m)) of 2.27 A(3) Da(-1) and a solvent content of 46%. Native X-ray data to 2.15 A resolution have been collected using synchrotron X-rays.

Crystallization↗

PubMed1999

Quantitative structure-activity relationship modeling of dopamine D(1) antagonists using comparative molecular field analysis, genetic algorithms-partial least-squares, and K nearest neighbor methods.

Several quantitative structure-activity relationship (QSAR) methods were applied to 29 chemically diverse D(1) dopamine antagonists. In addition to conventional 3D comparative molecular field analysis (CoMFA), cross-validated R(2) guided region selection (q(2)-GRS) CoMFA (see ref 1) was employed, as were two novel variable selection QSAR methods recently developed in one of our laboratories. These latter methods included genetic algorithm-partial least squares (GA-PLS) and K nearest neighbor (KNN) procedures (see refs 2-4), which utilize 2D topological descriptors of chemical structures. Each QSAR approach resulted in a highly predictive model, with cross-validated R(2) (q(2)) values of 0.57 for CoMFA, 0.54 for q(2)-GRS, 0.73 for GA-PLS, and 0.79 for KNN. The success of all of the QSAR methods indicates the presence of an intrinsic structure-activity relationship in this group of compounds and affords more robust design and prediction of biological activities of novel D(1) ligands.

Algorithms↗

PubMed1999

Antitumor agents. 194. Synthesis and biological evaluations of 4-beta-mono-, -di-, and -trisubstituted aniline-4'-O-demethyl-podophyllotoxin and related compounds with improved pharmacological profiles.

As a continuation of our structure-activity relationship studies, several new 4-beta-substituted 4'-O-demethyl-4-desoxypodophyllotoxins bearing mono-, di-, or trisubstituted anilines have been synthesized and evaluated as inhibitors of DNA topoisomerase II and tumor cell growth in tissue culture. Selected compounds were further evaluated as cytotoxic agents using a clonogenic survival assay. The target compounds include 4'-O-demethyl-4beta-[(4' '-(benzimidazol-2' '-yl)anilino]-4-desoxypodophyllotoxin (21), 4'-O-demethyl-4beta-(-)-(4' '-camphanamido-anilino)-4-desoxypodophyllotoxin (25), 4-beta-disubstituted-anilino-4'-demethyl-4-desoxypodophyllotoxins (18-20, 26), 4-alpha-disubstituted-anilino-4'-demethyl-4-desoxypodophyllotoxin (27), 4-beta-trisubstituted-anilino-4'-demethyl-desoxypodophyllotoxin (22, 23), and 4'-O-demethyl-4beta-[4' '-(benzimidazol-2' '-yl)amino]-4-desoxypodophyllotoxin (24). Among the target series, 19, 21, and 24 displayed significant growth inhibitory action against a panel of tumor cell lines including human epidermoid carcinoma of the nasopharynx (KB) and its etoposide-resistant (KB7B) and vincristine-resistant (vin20c KB) subclones, lung carcinoma (A549), human ileocecal carcinoma (HCT-8), human kidney carcinoma (CAKI-1), breast adenocarcinoma (MCF-7), and human malignant melanoma (SK-MEL-2) cells. Compounds 19, 21, 24, and 25 were "cleavable-complex"-forming DNA topoisomerase II inhibitors with either improved or similar activity compared with the prototype drug etoposide (VP-16). Compound 21 was the most active analogue, being 10-fold more potent than etoposide in both cell killing and topoisomerase II inhibition in vitro assays. Using mouse models of antitumor activity, 21 was effective against (P388/0) leukemia but not against the growth of a (MCF7) mammary tumor.

Animals↗

PubMed1999

Bioisosterism: interchange of 4-OH to 4-NH2 in vanillin or homovanillin ring of capsaicinoids.

A series of 4-amino Capsaicin analogs 15, 17 and 19 were prepared to investigate the bioisosteric effect of 4-amino group, and all these compounds exhibited moderate or weak potency from their analgesic test. From our previous results and others, 4-hydroxyl group as well as 3-methoxy substituent could be crucial for high analgesic activity. This biological result also shows that the activity is sensitive to alkyl chain length in hydrophobic region and the phenylacetic amides 19 are more active than the corresponding urea derivatives 17.

Analgesics, Non-Narcotic↗

PubMed1999

Crystallization and preliminary X-ray crystallographic analysis of deoxycytidylate hydroxymethylase from bacteriophage T4.

Deoxycytidylate hydroxymethylase from bacteriophage T4 is a homodimeric enzyme in which each polypeptide chain consists of 246 amino-acid residues. It has been crystallized in the presence of its substrate, deoxycytidine monophosphate, at room temperature using sodium citrate as precipitant. The crystals are monoclinic, belonging to space group C2, with unit-cell parameters a = 174.22, b = 53.12, c = 75.17 A, beta = 115.29 degrees. The asymmetric unit contains one homodimer, with a corresponding Vm of 2.65 A3 Da-1 and solvent content of 54%. Native diffraction data to 1.6 A resolution have been collected from two crystals using synchrotron radiation.

Bacteriophage T4↗

PubMed1999

Rational type of laparoscopic hysterectomy and safety in anesthetic profiles.

OBJECTIVE: To evaluate what type of surgery would be more reasonable among 3 types of laparoscopic hysterectomy and to evaluate the safety of cardio-pulmonary changes on these patients during these operations. METHOD: A retrospective study was carried out in 215 women who underwent laparoscopic hysterectomy including laparoscopic-assisted vaginal hysterectomy (LAVH), laparoscopic hysterectomy (LH), total laparoscopic hysterectomy (TLH). Blood gas analysis, end-tidal CO2 levels and vital signs were checked and compared with control and preceding values. RESULTS: The average duration of operation was 102.5 min, 83.8 min and 118.3 min for LAVH (n = 97), LH (n = 75) and TLH (n = 43), respectively (p < 0.05). The average amount of bleeding was 297.5 ml, 152.3 ml and 149.2 ml for each type of hysterectomy, respectively. Hemoglobin decreased by an average of 1.6 g/100 ml, 0.9 g/100 ml and 0.8 g/100 ml, respectively. There was a lesser amount of bleeding for LH and TLH than for LAVH (p < 0.05). Profiles of blood gas analysis and expiratory CO2 varied significantly according to the operative stages under controlled anesthesia (p < 0.05), but were within the normal range. CONCLUSION: These results demonstrate that laparoscopic procedures advancing below the uterine vasculature can be considered effective for hysterectomies and that proper anesthesia can safely control the cardio-pulmonary changes during laparoscopic hysterectomy.

Adult↗

PubMed1999

Long-term growth after hypophyseal stalk transection and hypophysectomy of beef calves.

Hypothalamic hormones regulate episodic and basal secretion of hormones from the anterior pituitary gland that affect metabolism and growth in cattle. This study focused on long-term growth in young calves subjected to hypophysectomy (HYPOX), hypophyseal stalk transection (HST), and sham operation control (SOC). Cross-bred (Hereford x Aberdeen Angus) and Hereford, and Aberdeen Angus calves were HYPOX (n = 5), HST (n = 5), or SOC (n = 8) at 146 +/- 2 days of age, whereas another group was HST (n = 5) or SOC (n = 7) at 273 +/- 5 days of age. Body weight was determined every 21 days from birth to 1008 days of age. Anterior vena cava blood was withdrawn at 4-day intervals from day 64-360 for RIA of GH, TSH, T4, T3, and LH, and at 20-min intervals for 480 min to determine episodic hormone secretion. Daily feed intake was determined in HST and SOC calves during an 80-day period. Birth weight averaged 35 +/- 1 kg (+/- SE) and was 142 +/- 4 kg at 126 days and 208 +/- 8 kg at 252 days before surgery. From day 146-1008, growth was arrested (P < 0.001) in HYPOX (0.06 +/- 0.01 kg/day) compared with SOC (0.50 +/- 0.04 kg/day) calves. Growth continued but at a significantly lower rate (P < 0.05) in calves HST at 146 days (0.32 +/- 0.07 kg/day) and 273 days (0.32 +/- 0.06 kg/day) compared with SOC (0.50 +/- 0.09 kg/day). Growth continued to be impaired to 1008 days, but more so in those HST at 146 days (432 +/- 43 kg BW) than 273 days (472 +/- 5 kg BW) and less (P < 0.05) than SOC (586 +/- 37 kg BW). Daily feed intake was consistently less (P < 0.05) in HST compared with SOC calves. Although episodic GH secretion was abolished and peripheral serum GH concentration remained consistently lower in HST (2.4 ng/ml) than SOC (5.5 ng/ml; P < 0.01), the calves continued to grow throughout 1008 days. Peripheral serum TSH concentration was less (P < 0.05) HST compared with SOC calves. There was an abrupt decrease (P < 0.001) in serum T4 (4-fold) and T3 (3-fold) concentration after surgery that remained to 360 days in HST compared with SOC calves. At the time calves were killed, pituitary gland weight was markedly reduced (P < 0.001) in HST (0.18 +/- 0.01 g/100 kg BW) compared with SOC (0.54 +/- 0.03 g/100 kg BW). Histological examination of pituitary glands from HST calves indicated the persistence of secretory GH and TSH cells in the same areas of the adenohypophysis as SOC calves. Coronal sections of the gland stained with performic acid-Alcian blue-periodic acid-Schiff-orange G, revealed GH and TSH secreting cells in HST calves similar to controls. These results indicate that long-term growth continues, but at a slower rate, after hypophyseal stalk transection of immature calves in spite of complete abolition of episodic GH secretion and consistently decreased basal secretion of GH, TSH, T4, and T3 compared with sham-operated animals. Growth was abolished after hypophysectomy of immature calves in which circulating GH and TSH was undetectable.

Animals↗

PubMed1998

Ionophores and receptors using cation-pi interactions: collarenes.

Cation-pi interactions are important forces in molecular recognition by biological receptors, enzyme catalysis, and crystal engineering. We have harnessed these interactions in designing molecular systems with circular arrangement of benzene units that are capable of acting as ionophores and models for biological receptors. [n]Collarenes are promising candidates with high selectivity for a specific cation, depending on n, because of their structural rigidity and well-defined cavity size. The interaction energies of [n]collarenes with cations have been evaluated by using ab initio calculations. The selectivity of these [n]collarenes in aqueous solution was revealed by using statistical perturbation theory in conjunction with Monte Carlo and molecular dynamics simulations. It has been observed that in [n]collarenes the ratio of the interaction energies of a cation with it and the cation with the basic building unit (benzene) can be correlated to its ion selectivity. We find that collarenes are excellent and efficient ionophores that bind cations through cation-pi interactions. [6]Collarene is found to be a selective host for Li+ and Mg2+, [8]collarene for K+ and Sr2+, and [10]collarene for Cs+ and Ba2+. This finding indicates that [10]collarene and [8]collarene could be used for effective separation of highly radioactive isotopes, 137Cs and 90Sr, which are major constituents of nuclear wastes. More interestingly, collarenes of larger cavity size can be useful in capturing organic cations. [12]Collarene exhibits a pronounced affinity for tetramethylammonium cation and acetylcholine, which implies that it could serve as a model for acetylcholinestrase. Thus, collarenes can prove to be novel and effective ionophores/model-receptors capable of heralding a new direction in molecular recognition and host-guest chemistry.

Benzene Derivatives↗

PubMed1998

Seasonal regulation of prolactin secretion in hypophyseal stalk transected beef calves.

Seasonal regulation of prolactin secretion was investigated in crossbred beef heifer calves. Calves were randomly assigned to hypophyseal stalk transection (HST, n = 6) or sham-operation control (SOC, n = 6) groups and fitted 1 day before surgery with an indwelling external jugular catheter. Prolactin (PRL), growth hormone (GH), thyroid stimulating hormone (TSH), thyroxine (T4), and tri-iodothyronine (T3) in peripheral serum were measured by radioimmunoassay in samples obtained before and after HST or SOC. During the first 8 days after HST, PRL concentrations remained significantly greater than SOC, but then decreased in both HST and SOC calves to 4 +/- 2 (+/- SE) and 10 +/- 3 ng/ml, respectively (P < 0.001). PRL remained low in both HST and SOC groups for three months after surgery. By four months, HST calves had lower basal PRL (5 +/- 1 ng/ml) than observed in SOC (40 +/- 4 ng/ml), and seasonal changes in PRL blood concentration also were attenuated by HST. Although HST reduced PRL secretion, it did not abolish the effect of seasonal changes (P < 0.01); circulating PRL concentration increased six-fold by shifts in photoperiod and temperature from winter to summer in these stalk-transected calves. The SOC group had higher serum GH during the winter (3.8 +/- 0.8) than in July (1.3 +/- 0.03 ng/ml). The HST group had the opposite profile of GH concentration, however, with concentrations being higher during May through July. Thyroid stimulating hormone secretion was partly sustained after stalk transection possibly by negative feedback of reduced circulating thyroxine and tri-iodothyronine. These results in both hypophyseal stalk-transected and sham-operated beef calves maintained in a natural environment strongly suggest that hypothalamic regulation of PRL secretion by adenohypophyseal cells is extremely sensitive to seasonal changes throughout the year. Additionally, immediately after HST, PRL blood concentration remains significantly greater than in SOC calves but eventually decreases to low blood concentration in HST calves, and unlike that seen after HST in primates. Regardless, basal PRL serum concentration responds to seasonal changes, but a less distinct change in basal GH serum concentration in HST calves than seen in the SOC calves.

Animals↗

PubMed1998

Antiporcine relaxin (antipRLX540) treatment decreases relaxin plasma concentration and disrupts delivery in late pregnant pigs.

Antibody against porcine relaxin (antipRLX540; 1:950,000) was produced in sheep and used to determine the effect on relaxin and progesterone secretion, and on parturition in late pregnant pigs. In group 1, Yorkshire gilts with normal estrous cycles were bred on the second observed estrus and fitted with an indwelling jugular cannula and an intraperitoneal cannula on day 100 of pregnancy. Gilts were infused at 6-h intervals with antipRLX540 (n = 10) or PBS (n = 10) beginning on day 103 until parturition. From days 103 to 120, daily blood samples (10 ml) were collected for RIA of relaxin, progesterone, and prolactin. In group 2, bred gilts were randomly assigned to antipRLX540 (n = 11), relaxin (n = 5), and PBS (n = 8) treatment on days 111, 113, and 115. Blood was collected twice daily from day 108 to 120, and every 20 min on days 111, 113, and 115 beginning 60 min before treatment and continuing 180 min. Parturition in gilts given antipRLX540 occurred on day 112.7 compared with day 114.0 in relaxin-treated gilts and day 114.3 in PBS controls (P < 0.05). Duration of delivery from first to last piglet was greatly delayed in antipRLX540 gilts (240 min) compared with PBS controls ([117 min] P < 0.005). Average number of stillborns was greater in antipRLX540- than in PBS-treated controls (2.4 vs. 1.0; P < 0.05). Relaxin concentration in peripheral plasma was lower in antipRLX540-treated gilts from day 105 to 110, but on day 113 the antipRLX540-treated group had a greater relaxin peak release compared with PBS-treated animals (P < 0.05). Plasma progesterone concentrations were similar in antipRLX540- and PBS-treated gilts throughout the period of the study. In group 2, by day 113, progesterone decreased in antipRLX540-treated gilts compared with relaxin- and PBS-treated gilts. Prolactin levels were similar in both antipRLX540- and PBS-treated gilts; however, from 1 to 3 days postpartum the antipRLX540 group had higher prolactin concentration (P < 0.05). The results indicate that antipRLX540 decreased circulating plasma concentrations of unbound or free relaxin during the last 10 days of pregnancy in Yorkshire gilts. AntipRLX540 markedly increased both the duration of delivery of piglets and the average number of stillbirths in this litter-bearing species compared with PBS-treated controls. This study provides strong evidence that increasing circulating concentrations of relaxin during late pregnancy is crucial for unimpaired parturition in the pig.

Animals↗

PubMed1998

Three-dimensional quantitative structure-activity relationship study of nonsteroidal estrogen receptor ligands using the comparative molecular field analysis/cross-validated r2-guided region selection approach.

A newly developed comparative molecular field analysis (CoMFA) technique, the cross-validated r2-guided region selection (CoMFA/q2-GRS) method, has been used to build a quantitative structure-activity relationship (3D-QSAR) for nonsteroidal estrogen receptor (ER) ligands. Ligands included in this study belong to a series of diethylstilbestrol (DES) and indenestrol analogues whose affinities for the mouse ER (mER) have been determined in our laboratory. The final model utilized 30 compounds and yielded a q2GRS (cross-validated r2, guided region selection) of 0.796, as compared to a q2 of 0.720 for conventional CoMFA, with a standard error of prediction of 0.594 at 3 principal components. This model was used to visualize steric and electrostatic features of the ligands that correspond with ER binding affinity. Results obtained from the CoMFA steric and electrostatic plots of this model have also been compared to information from the ER binding affinities of substituted estradiol analogues. This is in an effort to determine structural features of compounds in the CoMFA analysis that may correspond to those of the estradiol analogues and to further clarify the mode of binding of nonsteroidal ER ligands.

Animals↗

PubMed1998

Differential effects of dexamethasone and RU 486, an antigestagen and antiglucocorticoid, on progesterone and relaxin secretion in hysterectomized pigs with aging corpora lutea.

Pregnancy lasts about 114 days in pigs. Porcine corpora lutea produce not only progesterone but also relaxin (RLX), a peptide hormone that plays a critical role in suppressing uterine motility during pregnancy and in remodelling connective tissues in preparation for imminent parturition. Progesterone concentrations in peripheral blood remain elevated (approximately equal to 25 ng ml-1) for the major part of pregnancy and decrease just before parturition. The decrease in progesterone coincides with the peak prepartum RLX release. Glucocorticoid or antiglucocorticosteroid steroid, RU 486, administration during late pregnancy can induce parturition in the pig. Peak release of RLX and a coincident decrease of progesterone in the circulating blood can also occur in the complete absence of fetuses and uterus in the pig. The effects of glucocorticoid or antiglucocorticoidsteroid administration to such hysterectomized pigs on the secretion of RLX and progesterone were examined in this experiment. Unmated Yorkshire gilts were hysterectomized on days 6-8 (estrus = day 0) and given dexamethasone (total of 30 mg day-1; 2 times i.m. at 0800 and 1600 h) from days 110-118; control animals received vehicle injection during this period. RU 486 (4 mg kg1 body weight) was orally administered once daily (days 111-115) at 0800 h; placebo-treated controls were given the same amount of feed without the drug at this time. RLX concentrations in blood were markedly suppressed (P < 0.01) during dexamethasone treatment (3 +/- 0.9 ng ml-1; mean +/- S.E.) whereas a peak release of RLX (17 +/- 2.8 ng ml-1) occurred in the control group on day 113. In contrast, progesterone concentrations (14 +/- 2.5 ng ml-1) were unaffected by dexamethasone treatment compared with the controls (15 +/- 2.0 ng ml-1; P > 0.82). Upon withdrawal of dexamethasone on day 118, RLX plasma concentrations began to increase and peaked at 14 +/- 2.1 ng ml-1 (P < 0.01) on day 120. In contrast, the antiglucocorticosteroid given to hysterectomized pigs bearing aging corpora lutea caused a marked elevation in circulating levels of progesterone and delayed RLX release until after the end of drug treatment on day 115. Average weights of corpora lutea collected at day 12 were similar between treated and control groups (427 +/- 7 vs. 433 +/- 6 mg; P > 0.68). Results indicate that a synthetic glucocorticoid, dexamethasone, suppresses RLX secretion without causing luteolysis and such suppression is reversible; progesterone secretion remained unaffected. In contrast, the antiglucocorticoid, RU 486, raised progesterone plasma concentration and delayed RLX peak release but did not suppress it during treatment. This experiment provides further evidence that relaxin and progesterone secretion from aging corpora lutea of pigs are regulated through separate mechanisms, and adrenal glucocorticoids may be involved in such a regulation process.

Animals↗