Association between Degree of Anaplasia and Degree of Inflammation with the Expression of COX-2 in Feline Injection Site Sarcomas.

Feline injection site sarcomas (FISSs) are mesenchymal neoplasms that develop at the sites of delivery of vaccines or other injectable products. Vaccine adjuvants can trigger an intense and persistent inflammatory response that may lead to neoplastic transformation. The proinflammatory role of cyclo-oxygenase (COX)-2 is well known and its overexpression has prognostic value in multiple neoplastic processes. One hundred and seventeen FISSs were evaluated for the degree of inflammation and anaplasia. Immunohistochemistry was used to determine the expression of COX-2 in these sarcomas. There was a significant association between the degree of inflammation and the expression of COX-2 by neoplastic cells. COX-2 expression was lower in tumours with higher degrees of anaplasia. These findings may be useful in predicting the sensitivity of FISSs to treatment with COX-2 inhibitors. The potential therapeutic use of such agents could then be restricted to tumours with lower degrees of anaplasia.

Alteration of feedback mechanism of estrogen on gonadotropin by sulpiride-induced hyperprolactinemia.

Four normally cycling women received an iv injection of 20 mg Premarin (conjugated estrogens equivalent to 20 mg estrone sulfate) on the seventh day of two consecutive cycles; the second experiment was performed under sulpiride-induced hyperprolactinemia (mean PRL level = 906 microU MRC standard 71/222 per ml; significantly 7.8 times greater than mean control level of 115 microU MRC standard 71/222 per ml, P less than 0.001). In comparison to the control experiment, sulpiride-induced hyperprolactinemia prevented the occurrence of any gonadotropin peak within the 84 h of estrogen administration; the negative feedback effect of estrogen on gonadotropin secretion was maintained and was even potentiated. These alterations of feedback mechanisms of estrogen were considered to be related to hyperprolactinemia itself rather than to sulpiride. Five other normally cycling women received iv injections of 100 microgram LRH on the 22nd day of a cycle under sulpiride-induced hyperprolactinemia since the onset of menstruation. Their mean LH response was somewhat greater (although not statistically significant) and their mean FSH response was considerably greater (P less than 0.001 at all times) than those of a control group of 10 women tested in their luteal phase. The results of these LRH tests under sulpiride-induced hyperprolactinemia give some support to the concept that hyperprolactinemia interferes at the hypothalamic or higher level with cyclic release of indogenous LRH.

A simple and efficient system for the construction of phoA gene fusions in gram-negative bacteria.

We have developed a two-plasmid system for generating gene fusions between phoA and cloned genes encoding envelope proteins. The vector plasmid carries a temperature-sensitive replication system and can be rescued at high temperature by insertion of an IS1-based transposon carrying the ori region of pBR322 and a phoA gene lacking transcription and translation initiation signals. The vector plasmid also carries the transfer origin of the conjugative plasmid, F, permitting transfer into a suitable recipient strain. We have used this system in the analysis of the bla gene cloned from pBR322.

Characterization of two groEL genes in Streptomyces coelicolor A3(2).

Two Streptomyces coelicolor A3(2) groEL-like genes, groEL1 and groEL2, were cloned and characterized. Pulsed-field-gel electrophoresis located these genes, which were not adjacent, in the same segment of the chromosome. Nucleotide sequence analysis revealed that groEL1, but not groEL2, was preceded by a groES-like gene. Northern blots showed that heat shock induced the accumulation of transcripts corresponding to groES (0.7 kb), groES/EL1 (2.3 kb) and groEL2 (2.1 kb). Unique transcription start points and promoters were located for groES/EL1 and groEL2, having -10 and -35 hexamers similar to eubacterial vegetative promoters. Regions located 5' to the groES/EL1 or groEL2 structural genes contain 'GCACTCN9GAGTGC' motifs conserved upstream from the heat-shock genes of other bacteria.

P2Y- and P2U-mediated increases in internal calcium in single bovine aortic endothelial cells in primary culture.

Increases in intracellular calcium ([Ca2+]i) to ATP, ADP, AMP, adenosine, UTP, 2-methylthio ATP (2-MeSATP), 2-methylthio ADP (2-MeSADP) and alpha,beta-methylene ATP (alpha,beta-meATP) were investigated in single bovine aortic endothelial cells (BAEC) in primary culture using Indo-1. Evidence was obtained for the presence of P2Y and P2U, but not P2X receptors. Normalized concentration-effect curves for ATP, UTP and 2-MeSATP were biphasic in shape. At 10 nM, the agonist rank order was UTP > ATP approximately 2-MeSATP, while above 1 microM, it was ATP > or = UTP > or = 2-MeSATP. No cross-desensitization between responses to P2U and P2Y receptors was observed in normal external solution. However, when internal Ca2+ stores were depleted by exposure to 2-MeSATP or UTP in Ca2+-free solution and agonists then re-applied in presence of external Ca2+, homologous but not heterologous desensitization was seen. In the same conditions, heterologous desensitization was observed for UTP after ATP but not for ATP after UTP. Taken together, the results are consistent with the coexistence of P2Y and P2U receptors in primary-cultured BAEC and suggest that upon activation, different intracellular signaling pathways could be involved in increasing [Ca2+]i.

(E)-5-(Tributylstannylmethylidene)-5H-furan-2-ones: versatile synthons for the stereospecific elaboration of gamma-alkylidenebutenolide skeletons.

[reaction: see text] Stereoselective construction of (E)-gamma-tributylstannylmethylidene butenolides 1 was achieved through the palladium-catalyzed tandem cross-coupling/cyclization reactions of tributylstannyl 3-iodopropenoate derivatives with tributyltinacetylene. Iododestannylation of 1 occurs with inversion of the configuration of the exocyclic double bond while the observed selectivity in the Stille reaction was found to be dependent on the nature of the aryl halide.

Vasoactive intestinal peptide potentiates and directly stimulates catecholamine secretion from rat adrenal chromaffin cells.

The actions of vasoactive intestinal polypeptide (VIP) on catecholamine secretion and changes in [Ca2+]i in single rat chromaffin cells were studied using amperometry and Indo-1. Application of VIP prior to acetylcholine (ACh) or co-application of VIP and ACh enhanced secretion by 94% and 153% respectively, compared to ACh alone. [Ca2+]i was increased by 17% when VIP was preapplied and by 73% upon co-application. Exposure to VIP before stimulation with 60 mM K+ enhanced secretion by 68%, but not [Ca2+]i. VIP application prior to DMPP and nicotine had no effect on [Ca2+]i, but increased [Ca2+]i signals to muscarine by 18%. VIP co-application potentiated only [Ca2+]i responses to muscarine, by 28%. The effect of VIP on muscarine-induced [Ca2+]i signals was mimicked by 8-Br-cAMP, and both were blocked by H-89, a protein kinase A inhibitor. Long-lasting increases in secretion accompanied by a sustained rise in [Ca2+]i to VIP alone were seen in 55% of cells. Removal of Ca2+ or addition of La3+ inhibited both responses, while L-, N- and P-type Ca2+ channel blockers were ineffective. SK&F 96365 inhibited VIP-induced secretion completely and rises in [Ca2+]i by 75%. Neither 8-Br-cAMP nor 8-Br-cGMP evoked responses similar to VIP alone. Thus in rat chromaffin cells, VIP acts both directly as a neurotransmitter in provoking sustained catecholamine secretion in a cAMP-independent manner, and also by enhancing ACh-induced secretion, via a cAMP-dependent action involving muscarinic receptors.

Palladium-catalyzed cross-coupling of 3-iodobut-3-enoic acid with organometallic reagents. Synthesis Of 3-substituted but-3-enoic acids

3-Substituted but-3-enoic acids were obtained in good yields under mild experimental conditions by palladium-catalyzed cross-coupling of 3-iodobut-3-enoic acid with organozinc or organotin compounds using PdCl(2)(MeCN)(2) as catalyst and DMF as solvent.

A long-term nutrition intervention experience: lipid responses and dietary adherence patterns in the Multiple Risk Factor Intervention Trial.

Nutrition counselors in the Multiple Risk Factor Intervention Trial (MRFIT) were able to help middle-aged men who were at high risk for coronary heart disease change their dietary habits, maintain those changes over time, and decrease their serum cholesterol levels. Most of a 7.5% mean serum cholesterol reduction achieved after 6 years of nutrition intervention occurred during the first year of the trial and was thereafter sustained. Total cholesterol and low-density lipoprotein cholesterol fraction decreases indicated improvement in terms of coronary heart disease risk. The food record rating, a numerical, semi-objective adherence technique that assesses a 3-day food record with respect to lipid-lowering potential, was used throughout the trial to measure adherence to recommended food patterns. Participants with lower food record rating scores, which indicate better adherence, demonstrated greater reductions in serum total cholesterol, plasma total cholesterol, and low-density lipoprotein fraction cholesterol determinations on a group basis. Subjective evaluations of the suitability of home and working environments, evidence of deviation from the MRFIT food patterns, and overall nutrition program motivation also showed that as ratings in each category became more favorable, lower food record rating scores and greater blood lipid reductions were consistently observed. The subgroup of participants who were non-smokers and not hypertensive demonstrated greater lipid responses and better dietary adherence. Continued smoking and antihypertensive medications appeared to adversely influence dietary adherence and/or lipid reductions. The MRFIT experience, however, demonstrated for the first time that dietary changes and blood lipid reductions can be achieved after the initial intervention effect, despite a continued emphasis on high blood pressure management and smoking cessation.

Comparative study of oral and written picture description in patients with Alzheimer's disease.

Oral and written picture descriptions were compared in 22 patients with Alzheimer's disease (AD) and 24 healthy elderly subjects. AD patients had a significant reduction of all word categories, which, similarly to controls, was more pronounced in written than in oral texts. They also reported fewer information units than controls, but without task difference. At the syntactic level, written descriptions of AD subjects were characterized by a diminution of subordinate clauses and a reduction of functors. More grammatical errors were present in written descriptions by AD and control subjects. AD and control groups produced an equivalent number of semantic errors in both tasks. However, in oral description, AD patients had more word-finding difficulties. In sum, AD descriptions were always shorter and less informative than control texts. Additionally, written descriptions of AD patients appeared shorter and more syntactically simplified than, but as informative as oral descriptions. Whereas no phonemic paraphasias were observed in either group, AD patients produced many more graphemic paragraphias than controls produced. Furthermore, written descriptions had more irrelevant semantic intrusions. Thus, as compared to oral descriptions, written texts appeared to be a more reliable test of semantic and linguistics difficulties in AD.

Toxocara canis in experimentally infected pigs: migratory pattern and tissue lesions.

Fifteen Yorkshire female pigs were inoculated with 100,000 infective T. canis eggs. Three animals were used as uninfected controls. Groups of three infected pigs were euthanized by accepted methods on days 7, 14, 21, 28 and 126 p.i., respectively. Larvae were recovered from all animals included in each group slaughtered on days 7 and 14 p.i.; on day 21 p.i. from two pigs, on day 28 p.i. from one, and no larvae were found on day 126 p.i. Differences in the mean number of larvae per gram in lymph nodes, liver and lungs between slaughter days, were significant for livers on day 7 p.i. and for lungs on day 14 p.i. (P < 0.10). The decrease over time was significant in all the organs that previously had larvae. Larvae were not found in the other organs and tissues analysed. Macroscopical lesions were found in the liver, lungs and lymph nodes on days 7, 14, 21, and 28 p.i. The entire surface of the liver was covered with small white spots on day 7 p.i., on days 14 and 21 p.i. the spots were distinctly nodular and, in some places, individual lesions were confluent. Lesions had apparently started to heal on days 28 and 126 p.i. appearance was normal. Lymph nodes were enlarged and oedematous during the first 4 weeks and the lungs had small areas of consolidation visible all over the surface, but by day 126 p.i., no visible lesions could be seen. Microscopical lesions were observed in the liver on day 7 p.i., with a largely periportal hepatitis. Numerous eosinophils and lymphocytes were present. The typical granulomatous reaction was observed on days 14 and 21 p.i. with a central necrotic core and a narrow region of fibroblastic tissue. By day 28 p.i. lesions had almost disappeared and the number of eosinophils was fewer. There were fewer leukocytes and the fibrous tissue had disappeared from the liver on day 126 p.i. For the first 3 weeks, pictures of the lymph nodes and the lungs were characterised by the formation of a granuloma. In the center of the granuloma larvae were observed. The majority of the lesions had healed by day 126 p.i.

Termination of pregnancy in dogs by oral administration of dexamethasone.

Dexamethasone was administered orally for 7.5 or 10 d to each of 20 pregnant bitches beginning at an estimated 28 to 51 d of gestation, using 1 of 2 dose regimens. Five bitches were given dexamethasone 3 times a day for 10 d, with the highest dose of 0.2 mg/kg for 5 d and then at progressively decreasing doses of 0.16-0.02 mg/kg for 5 d. The 15 remaining bitches were given dexamethasone 2 times a day for 7.5 d, increasing from 0.1 to 0.2 mg/kg over the first 3 administrations, then remaining at 0.2 mg/kg on Days 2 to 5, and decreasing from 0.16 to 0.02 mg/kg over the last 5 administrations. The side effects, including mild polydipsia and polyuria, disappeared when treatment was discontinued. Depending on the stage of pregnancy, uterine contents were either resorbed or aborted, or both. Pregnancy was terminated within 2 to 16 d after the start of treatment in all treated bitches, at 2 to 5 d of treatment in 2 of 3 bitches treated at 40 to 51 d of pregnancy, and at 0 to 4 d after the end of treatment in most of the 17 bitches treated at 28 to 35 days of pregnancy. Oral administration of dexamethasone appears to be a potentially useful pharmacologic treatment for the termination of unwanted pregnancy in the bitch.

Uveodermatological syndrome (Vogt-Koyanagi-Harada-like syndrome) with generalized depigmentation in a Dachshund.

A 3-year-old, female, black and tan Dachshund was referred with visual impairment, bilateral anterior and posterior uveitis, poliosis, and generalized dermal depigmentation. Complete blood counts and biochemical parameters, including T3 and T4, were normal. The skin biopsy showed lichenoid dermatoses with dermal infiltration of histiocytes and lymphocytes suggesting uveodermatological syndrome. Medical treatment was initiated with oral prednisone and azathioprine, and topical prednisolone and atropine. The clinical signs improved, vision was retained, and the skin began to repigment 2 months following the initiation of therapy.

[Analysis of mechanisms whereby pharmacologically-induced hyperprolactinemia leads to altered gonadotropin secretion (author's transl)]. / Analyse des mécanismes d'altération de la fonction gonadotrope par l'hyperprolactinémie pharmacologique.

Chronic hyperprolactinaemia was induced in 10 women since the onset of a cycle for 2 or 3 consecutive cycles by administration of sulpiride (3 X 50 mg/day) or tiapride (100-200 mg/day). The resulting perturbations of the menstrual cycle included short luteal phase cycles, anovulatory cycles and amenorrhoea. The analysis of the hormonal profiles under treatment, as compared to the control cycles, indicate an impact of hyperprolactinaemia both preferentially at the hypothalamic (alteration of the secretion or endogenous LRH) and accessorily at the ovarian (lack of progesterone secretion by the near mature follicle) levels.

.VO2 kinetics during supramaximal exercise: relationship with oxygen deficit and 800-m running performance.

The aim of this study was to compare .VO2 kinetics of highly- versus recreationally-trained subjects during a constant velocity test of supramaximal intensity. Eighteen trained male subjects were recruited to one of two groups: highly trained (HT, n = 8, .VO(2max) = 70.1 +/- 6.5 ml . min (-1) . kg (-1)) and recreationally trained (RT, n = 10, .VO(2max) = 63.2 +/- 6.4 ml . min (-1) . kg (-1)). All subjects performed an incremental test to exhaustion for the determination of .VO(2max) and peak treadmill velocity (PTV), two constant velocity tests at 110 % of PTV to determine .VO2 kinetics and oxygen deficit (O(2)def), and a 800-m time trial to determine running performance (mean velocity over the distance, V (800 m)). We found significant differences between HT and RT for the on-transient of the .VO2 response (tau, 24.7 +/- 3.3 and 30.9 +/- 7.0 s, respectively), the amplitude of the .VO2 response (60.0 +/- 5.0 and 53.5 +/- 5.7 ml . min (-1) . kg (-1), respectively) and V (800 m) (6.27 +/- 2.1 and 5.45 +/- 0.38 m . s (-1), respectively). O(2)def (24.6 +/- 2.7 and 27.7 +/- 7.8 ml . kg (-1), respectively) and the gain of the .VO2 response (193 +/- 14 and 194 +/- 13 ml . kg (-1) . m (-1), respectively) were similar between groups. tau was associated with O(2)def (r = 0.90, p < 0.05), but not with V (800 m) (r = 0.30, p > 0.05). It was concluded that HT subjects exhibited faster on-kinetics and higher amplitude than their RT counterparts. The higher amplitude was not thought to reflect any difference in underlying physiological mechanisms. The faster tau, whose exact mechanisms remain to be elucidated, may have practical implications for coaches.

Anaerobic running capacity determined from a 3-parameter systems model: relationship with other anaerobic indices and with running performance in the 800 m-run.

The purpose of this study was to compare anaerobic running capacity (ARC, i.e., the distance that can be run using only stored energy sources in the muscle) determined from a 3-parameter systems model with other anaerobic indices and with running performance in the 800 m. Seventeen trained male subjects (.VO(2max) = 66.54 +/- 7.29 ml . min (-1) . kg (-1)) performed an incremental test to exhaustion for the determination of .VO(2max) and peak treadmill velocity (PTV), five randomly ordered constant velocity tests at 95, 100, 105, 110, and 120 % of PTV to compute ARC and oxygen deficit (O(2)def, at 110 % of PTV), and a 800-m time trial to determine running performance (mean velocity over the distance, V (800 m)) and peak blood lactate concentration ([La (-)] (b, peak)). ARC (467 +/- 123 m) was positively correlated with O(2)def (56.35 +/- 18.47 ml . kg (-1); r = 0.57; p < 0.05), but not with [La (-)] (b, peak) (15.08 +/- 1.48 mmol . l (-1); r = - 0.16; p > 0.05). The O(2) equivalent of ARC (i.e., the product of ARC by the energy cost of running; 103.74 +/- 28.25 ml . kg (-1)), which is considered as an indirect estimation of O(2)def, was significantly higher than O(2)def (p < 0.01, effect size = 1.99). It was concluded that ARC is partially determined by anaerobic pathway, but that it probably does not provide an accurate measure of anaerobic capacity, if, however, O(2)def can be considered as a criterion measure for it.

Complete assignments of the 1H and 13C NMR spectra of 9,10-didehydro retinoic acids.

A new class of retinoic acids was synthesized containing a 9,10-rigid bond. 1H and 13C NMR spectra were assigned for eight new compounds (all-E, 13Z) containing a carboxylic acid or tertiobutylester polar end group. Assignments were based on the combination of one- and two-dimensional experiments (APT, COSY, HMBC).

On the role of cyclic AMP and the Fnr protein in Escherichia coli growing anaerobically.

The role of adenosine 3',5'-monophosphate (cAMP) and of the Fnr protein, a transcriptional regulator of anaerobic electron transport, in the expression of anaerobic respiration of Escherichia coli was investigated. Under conditions of fermentation or anaerobic respiration intracellular cAMP was formed in concentrations up to 4.6 nmol/g protein. From the enzymes of the anaerobic electron transfer chain from glycerol-3-P to fumarate only the expression of glycerol-3-P dehydrogenase (Freedberg WB, Lin ECC (1973) J Bacteriol 115:816-823), but not that of fumarate reductase required cAMP. Isolated Fnr protein, which has been suggested to be an additional site of action of cAMP under anaerobic conditions did not bind cAMP. It is concluded that cAMP in anaerobic growth like in aerobic growth acts as the effector of CRP and that catabolite repression plays an important regulatory role in anaerobic catabolism. The Fnr protein was present in constant amounts (0.06 mg/g cellular protein) and in constant molar mass (Mr 30,000) in aerobically and in anaerobically grown bacteria. This result excluded regulation of the activity of the Fnr protein by a change of concentration or by processing of the protein.