Stereospecific synthesis and structure-activity relationships of unsymmetrical 4,4-diphenylbut-3-enyl derivatives of nipecotic acid as GAT-1 inhibitors

Two complementary stereospecific synthetic approaches for the preparation of unsymmetrical ortho-substituted N-(4,4-diphenylbut-3-enyl) derivatives of nipecotic acid are described. Determination of the activity of the prepared compounds at the GAT-1 transporter highlighted differing SAR requirements of the E- and Z-phenyl rings, and led to the discovery of a compound with comparable potency to tiagabine. Some attempts to replace nipecotic acid with alternative novel amino acids are also described.     

Spatio‐temporal variations of Anopheles coluzzii and An. gambiae and their Plasmodium infectivity rates in Lobito,Angola

From 2003 to 2007, entomological surveys were conducted in Lobito town (Benguela Province, Angola) to determine which Anopheles species were present and to identify the vectors responsible for malaria transmission in areas where workers of the Sonamet Company live. Two types of surveys were conducted: (1) time and space surveys in the low and upper parts of Lobito during the rainy and dry periods; (2) a two‐year longitudinal study in Sonamet workers' houses provided with long‐lasting insecticide‐treated nets (LLIN), “PermaNet,” along with the neighboring community. Both species, An. coluzzii (M molecular form) and An. gambiae (S molecular form), were collected. Anopheles coluzzii was predominant during the dry season in the low part of Lobito where larvae develop in natural ponds and temporary pools. However, during the rainy season, An. gambiae was found in higher proportions in the upper part of the town where larvae were collected in domestic water tanks built near houses. Anopheles melas and An. listeri were captured in higher numbers during the dry season and in the low part of Lobito where larvae develop in stagnant brackish water pools. The infectivity rates of An. gambiae s.l. varied from 0.90% to 3.41%.     

Chronic Type A aortic dissection: could surgical intervention be guided by molecular markers?

Aortic dissection, occurring following a separation of the layers constituting the complex vascular walls, leads to the formation of a 'false' lumen and disrupts the regulation of aortic wall homeostasis and function. This clinical condition still represents an important health problem and is associated with high mortality. Its natural history mandates surgical intervention when exceeding 55 mm in diameter and involving the ascending portion of the aorta (Type A), on the bases of an anatomical classification dated back to 1965. An intriguing question rising is whether a dissection that overcomes that critic acute phase has still the indication to surgical intervention. Molecular analysis of chronic dissected aortic walls could help in understanding how morphology and structure are affected and whether tissue homeostasis is re-established. Thus, pursued by this consideration, we made a histological and immunohistochemical characterization of a chronic Type A dissection, reporting three major findings: endothelial cells line the aortic primitive lumen, as well as the 'false' one; walls of primitive and 'false' lumina are comparable in thickness; vascular layers in the 'false' lumen are made up of terminally differentiated cells. This evidence obtained in a single specimen encourages a meditation on the compulsory indication for surgical intervention.     

Effects of FSH and LH on ovarian and follicular blood flow, follicular growth and oocyte developmental competence in young and old mares

Objectives of the experiment were to determine the effects of mare age and gonadotropin treatments on dominant follicle vascularity, ovarian blood flow and dominant follicle growth and to associate follicular vascularity with oocyte developmental capacity. Growing follicles >30mm from young (4-9 years) and old (>20 years) mares were assessed for blood flow using color Doppler ultrasonography before maturation induction with recombinant equine LH (eLH) and immediately prior to oocyte collection at 20-24h after eLH. Pulsed Doppler was used to obtain resistance indices of ovarian arteries ipsilateral to preovulatory follicles. For eFSH-treated estrous cycles, eFSH administration was started after detection of a cohort of follicles ≥20 to <25mm and continued until a follicle >30mm. Oocytes were harvested using transvaginal, ultrasonic-guided aspirations and cultured and injected with sperm at 40±1h after eLH. Presumptive zygotes were incubated, and rates of cleavage (≥2 cells) and blastocyst formation were obtained. Embryos were transferred nonsurgically into recipients' uteri, and pregnancy rates were assessed. Vascularity (number of color pixels per total pixels) was higher (P=0.003) in the follicles of old compared to young mares, with no significant interaction of eFSH or eLH. Effects of eFSH and time from eLH on follicle vascularity were not significant. The vascularity of follicles associated with oocytes that did compared to those that did not form blastocysts was greater (P=0.048), although follicular vascularity was less (P=0.02) for follicles associated with oocytes that did compared to those that did not develop into pregnancies. Resistance indices were not different for age, eFSH treatment, time after eLH administration and oocyte developmental potential. Growth of the dominant follicle was not associated with vascularity, although advanced age tended (P=0.09) to have a negative effect on follicle growth.     

Vitrification of early-stage bovine and equine embryos

The objectives of this study were to: (1) determine an optimal method and stage of development for vitrification of bovine zygotes or early embryos; and (2) use the optimal procedure for bovine embryos to establish equine pregnancies after vitrification and warming of early embryos. Initially, bovine embryos produced by in-vitro fertilization (IVF) were frozen and vitrified in 0.25 mL straws with minimal success. A subsequent experiment was done using two vitrification methods and super open pulled straws (OPS) with 1- or 8-cell bovine embryos. In Method 1 (EG-O), embryos were exposed to 1.5 M ethylene glycol (EG) for 5 min, 7 M ethylene glycol and 0.6 M galactose for 30 s, loaded in an OPS, and plunged into liquid nitrogen. In Method 2 (EG-DMSO), embryos were exposed to 1.1 M ethylene glycol and 1.1 M dimethyl sulfoxide (DMSO) for 3 min, 2.5 M ethylene glycol, 2.5 M DMSO and 0.5 M galactose for 30 s, and loaded and plunged as for EG-O. Cryoprotectants were removed after warming in three steps. One- and eight-cell bovine embryos were cultured for 7 and 4.5 d, respectively, after warming, and control embryos were cultured without vitrification. Cleavage rates of 1-cell embryos were similar (P > 0.05) for vitrified and control embryos, although the blastocyst rates for EG-O and control embryos were similar and higher (P < 0.05) than for EG-DMSO. The blastocyst rate of 8-cell embryos was higher (P < 0.05) for EG-O than EG-DMSO. Therefore, EG-O was used to cryopreserve equine embryos. Equine oocytes were obtained from preovulatory follicles. After ICSI, injected oocytes were cultured for 1-3 d. Two- to eight-cell embryos were vitrified, warmed and transferred into recipient's oviducts. The pregnancy rate on Day 20 was 62% (5/8) for equine embryos after vitrification and warming. In summary, a successful method was established for vitrification of early-stage bovine embryos, and this method was used to establish equine pregnancies after vitrification and warming of 2- to 8-cell embryos produced by ICSI.     

Age and pasture effects on vernal transition in mares

The objectives of the present study were to determine if follicular activity was less in old than in young mares during the spring transition and if green pasture would hasten onset of the ovulatory season. Experiments were conducted over 2 sequential years using young mares (3 to 7 yr) and old mares (> or =14 yr). In Experiment 1, growth of the largest and second-largest follicles were compared for young mares (5 to 7 yr) and old mares (> or =14 yr) for 21 d prior to the first ovulation of the year. More follicular activity was noted in young than in old mares. Main effect of age was significant for diameter of the largest follicle, and interaction of day-by-age was significant for diameter of the second-largest follicle. Prior to the beginning of the breeding season, the mares were randomly divided into dry-lot and pasture groups. The interval from May 2 to ovulation was shorter (P < 0.005) for mares put on pasture on May 2 than for mares kept in dry lot (means +/- SEM, 14.5 +/- 2.7 and 21.3 +/- 3.2 d, respectively). In Experiment 2, follicular activity was compared among 3 age groups (3 to 7, 17 to 19, and > or =20 yr). The total number of follicles > or =10 mm was higher (P < 0.05) for young mares and lower (P < 0.05) for old mares than for mares of an intermediate age. Main effect of age and interaction of day-by-age were significant for diameter of largest and second-largest follicles, being smaller for mares > or =20 yr than for younger mares. The interval from development of a follicle > or =30 mm to ovulation was shorter (P < 0.05) for mares placed on pasture when a > or =30 mm follicle developed than the interval for mares kept in dry lot (5.7 +/- 0.7 and 8.2 +/- 0.9 d, respectively). In summary, less follicular activity occurred in old than in young mares during the transitional period, and mares pastured on green grass ovulated sooner in the spring than mares housed on dry lot and fed hay.     

Determination of argininosuccinate lyase and arginase activities with an amino acid analyzer

The measurement of argininosuccinate lyase (ASase) and arginase, both in liver and erythrocytes, was developed by using a commercial amino acid analyzer. The method is based upon the use of two different substrates, argininosuccinate and arginine for ASase and arginase, respectively, and the measurement of only one final metabolite: ornithine. The use of ornithine as a marker of biological activity of ASase is related to the fact that in the urea cycle, the specific activity of arginase is much higher than that of ASase; thus, during in vitro determinations, arginine, which is the product of ASase, is rapidly converted to ornithine. The sensitivity of the methods is very high since we were able to detect both activities using very diluted rat liver homogenates (0.10 mg protein/ml) or few microliters of human blood. In rat liver the Vmax for ASase and arginase were respectively 0.54 and 140 mumol/h/mg protein; the apparent Km values 1.25 and 13.5 mM. In human erythrocytes the Vmax for the same enzymes were 7.2 and 170 nmol/h/mg Hb and the apparent Km values were 0.66 and 9.5 mM. In 10 healthy volunteers the specific activity of ASase and arginase determined in blood were respectively 8.60 +/- 0.46 and 124.1 +/- 14.5 nmol/h/mg Hb. The results obtained from 2 patients suffering from argininosuccinic aciduria were also reported. In these latter cases while ASase was not detectable in blood, arginase activity was at the lowest end of the confidence limits determined in healthy volunteers.     

Plant communities and phytogeographical position of a large depression in the Great Chaco,Argentina

A survey is presented of the vegetation of the central region of the Santafesinian Chaco (Argentina), a scarcely populated flat area of 20 000 km², with seasonal flooding. Soils have a strong halo-hydromorphic character and vegetation is basically halophilous. Trees are scarce and most communities are savannas, grasslands or swampy vegetation. Twenty-three communities are described, some of them with several variants. The most widespread communities areSpartina argentinensis grasslands,Elyonurus muticus savannas and a complex of hygrophilous communities. The most important communities are distributed in relation to a topographical gradient, and their structure is shaped by recurrent flooding and fire disturbance. Most of the area is virgin land with very little human interference. The phytogeographical position of the area is discussed. Nomenclature: Burkart, A. 1969, 1974, 1979. Flora ilustrada de Entre Rios, INTA, Buenos Aires, and Cabrera, 1963, 1965a, b, 1967, 1968, 1970. Flora de la Provincia de Buenos Aires, INTA, Buenos Aires.