Frequency of Toxoplasma gondii antibodies in tufted capuchin monkeys (Cebus apella nigritus) from an ecological station in the State of São Paulo, Brazil

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii, an obligate intracellular parasite protozoan. A large percentage of animals presents specific antibodies caused by a previous exposition, resulting in a chronic infection. Felides are the definitive hosts and the other warm-blooded animals, including primates, are the intermediate hosts. This study was aimed to determine the prevalence of T. gondii infection in free-living tufted capuchin monkeys (Cebus apella nigritus) from an ecological station located on Mata de Santa Teresa, Ribeirão Preto, SP, Brazil. T. gondii antibodies were analyzed by modified agglutination test (MAT) in serum samples of 36 tufted capuchin monkeys, considering eight as cut-off titer. From the studied animals, 3/36 (8.33%; CI95% 3.0-21.9%) presented T. gondii antibodies, all with titer 32. No significative difference was observed relating to the sex (1/3 male and 2/3 female), and to the age (1/3 young and 2/3 adult) (P>0.05). Thus, these results demonstrate the presence of T. gondii antibodies in primates from São Paulo state.

Occurrence of Toxoplasma gondii DNA in sheep naturally infected and slaughtered in abattoirs in Pernambuco, Brazil

The aim of the present study was to assess the occurrence of antibodies to Toxoplasma gondii and to detect genomic DNA of the parasite in the reproductive organs, fetuses and fetal membranes of sheep in slaughterhouses in the state of Pernambuco, Brazil. The Indirect Immunofluorescence technique (IFA) was used for screening. The Polymerase Chain Reaction (PCR) was used to detect DNA of T. gondii in the animals that were positive in the serology. In the serology, 13/50 samples were positive and genomic DNA of T. gondii was detected in one uterus, tube, ovary, placenta and fetus (heart, brain and umbilical cord) sample from a sheep that was positive in the serology. The present study provides evidence of the occurrence of T. gondii DNA in the organs of the reproductive system, placenta and fetus of a naturally infected sheep.

Occurrences of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from four districts of Tocantins state, Brazilian Legal Amazon Region

Toxoplasmosis and neosporosis have been recognized as economically important diseases with considerable impact on the livestock industry. Little is known concerning the occurrence of Toxoplasma gondii and Neospora caninum in sheep from Tocantins state, Brazil. Here, we investigated antibodies against these parasites and associated factors in 182 sheep from Araguaína, Santa Terezinha do Tocantins, Arguianópolis and Palmeiras do Tocantins districts, Tocantins. Sheep sera were assayed for T. gondii and N. caninum IgG antibodies by indirect fluorescence antibody test (IFAT), using cut-off point at a dilution of 1:40 and 1:25 respectively. The prevalence of seropositive animal for T. gondii was 13.74% and 13.74% for N. caninum. None of the characteristics studied including reproductive problems, presence of cats, presence of dogs and veterinary care (p>0.05) was associated with occurrence of T. gondii or N. caninum infection. Only breed was identified as associated factor for the occurrence of toxoplasmosis in sheep (p<0.05). The present study is the first report on serum occurrence of T. gondii and N. caninum in sheep from the state of Tocantins, Brazil.

The supermassive binary black hole system OJ287

Abstract This doctoral thesis concerns the active galactic nucleus (AGN) most often referred to with the catalogue number OJ287. The publications in the thesis present new discoveries of the system in the context of a supermassive binary black hole model. In addition, the introduction discusses general characteristics of the OJ287 system and the physical fundamentals behind these characteristics. The place of OJ287 in the hierarchy of known types of AGN is also discussed. The introduction presents a large selection of fundamental physics required to have a basic understanding of active galactic nuclei, binary black holes, relativistic jets and accretion disks. Particularly the general relativistic nature of the orbits of close binaries of supermassive black holes is explored with some detail. Analytic estimates of some of the general relativistic effects in such a binary are presented, as well as numerical methods to calculate the effects more precisely. It is also shown how these results can be applied to the OJ287 system. The binary orbit model forms the basis for models of the recurring optical outbursts in the OJ287 system. In the introduction, two physical outburst models are presented in some detail and compared. The radiation hydrodynamics of the outbursts are discussed and optical light curve predictions are derived. The precursor outbursts studied in Paper III are also presented, and tied into the model of OJ287. To complete the discussion of the observable features of OJ287, the nature of the relativistic jets in the system, and in active galactic nuclei in general, is discussed. Basic physics of relativistic jets are presented, with additional detail added in the form of helical jet models. The results of Papers II, IV and V concerning the jet of OJ287 are presented, and their relation to other facets of the binary black hole model is discussed. As a whole, the introduction serves as a guide, though terse, for the physics and numerical methods required to successfully understand and simulate a close binary of supermassive black holes. For this purpose, the introduction necessarily combines a large number of both fundamental and specific results from broad disciplines like general relativity and radiation hydrodynamics. With the material included in the introduction, the publications of the thesis, which present new results with a much narrower focus, can be readily understood. Of the publications, Paper I presents newly discovered optical data points for OJ287, detected on archival astronomical plates from the Harvard College Observatory. These data points show the 1900 outburst of OJ287 for the first time. In addition, new data points covering the 1913 outburst allowed the determination of the start of the outburst with more precision than was possible before. These outbursts were then successfully numerically modelled with an N-body simulation of the OJ287 binary and accretion disc. In Paper II, mechanisms for the spin-up of the secondary black hole in OJ287 via interaction with the primary accretion disc and the magnetic fields in the system are discussed. Timescales for spin-up and alignment via both processes are estimated. It is found that the secondary black hole likely has a high spin. Paper III reports a new outburst of OJ287 in March 2013. The outburst was found to be rather similar to the ones reported in 1993 and 2004. All these outbursts happened just before the main outburst season, and are called precursor outbursts. In this paper, a mechanism was proposed for the precursor outbursts, where the secondary black hole collides with a gas cloud in the primary accretion disc corona. From this, estimates of brightness and timescales for the precursor were derived, as well as a prediction of the timing of the next precursor outburst. In Paper IV, observations from the 2004–2006 OJ287 observing program are used to investigate the existence of short periodicities in OJ287. The existence of a _50 day quasiperiodic component is confirmed. In addition, statistically significant 250 day and 3.5 day periods are found. Primary black hole accretion of a spiral density wave in the accretion disc is proposed as the source of the 50 day period, with numerical simulations supporting these results. Lorentz contracted jet re-emission is then proposed as the reason for the 3.5 day timescale. Paper V fits optical observations and mm and cm radio observations of OJ287 with a helical jet model. The jet is found to have a spine–sheath structure, with the sheath having a much lower Lorentz gamma factor than the spine. The sheath opening angle and Lorentz factor, as well as the helical wavelength of the jet are reported for the first time. Tiivistelmä Tässä väitöskirjatutkimuksessa on keskitytty tutkimaan aktiivista galaksiydintä OJ287. Väitöskirjan osana olevat tieteelliset julkaisut esittelevät OJ287-systeemistä saatuja uusia tuloksia kaksoismusta-aukkomallin kontekstissa. Väitöskirjan johdannossa käsitellään OJ287:n yleisiä ominaisuuksia ja niitä fysikaalisia perusilmiöitä, jotka näiden ominaisuuksien taustalla vaikuttavat. Johdanto selvittää myös OJ287-järjestelmän sijoittumisen aktiivisten galaksiytimien hierarkiassa. Johdannossa käydään läpi joitakin perusfysiikan tuloksia, jotka ovat tarpeen aktiivisten galaksiydinten, mustien aukkojen binäärien, relativististen suihkujen ja kertymäkiekkojen ymmärtämiseksi. Kahden toisiaan kiertävän mustan aukon keskinäisen radan suhteellisuusteoreettiset perusteet käydään läpi yksityiskohtaisemmin. Johdannossa esitetään joitakin analyyttisiä tuloksia tällaisessa binäärissä havaittavista suhteellisuusteoreettisista ilmiöistä. Myös numeerisia menetelmiä näiden ilmiöiden tarkempaan laskemiseen esitellään. Tuloksia sovelletaan OJ287-systeemiin, ja verrataan havaintoihin. OJ287:n mustien aukkojen ratamalli muodostaa pohjan systeemin toistuvien optisten purkausten malleille. Johdannossa esitellään yksityiskohtaisemmin kaksi fysikaalista purkausmallia, ja vertaillaan niitä. Purkausten säteilyhydrodynamiikka käydään läpi, ja myös ennusteet purkausten valokäyrille johdetaan. Johdannossa esitellään myös Julkaisussa III johdettu prekursoripurkausten malli, ja osoitetaan sen sopivan yhteen OJ287:n binäärimallin kanssa. Johdanto esittelee myös relativististen suihkujen fysiikkaa sekä OJ287- systeemiin liittyen että aktiivisten galaksiydinten kontekstissa yleisesti. Relativististen suihkujen perusfysiikka esitellään, kuten myös malleja kierteisistä suihkuista. Julkaisujen II, IV ja V OJ287-systeemin suihkuja koskevat tulokset esitellään binäärimallin kontekstissa. Kokonaisuutena johdanto palvelee suppeana oppaana, joka esittelee tarvittavan fysiikan ja tarpeelliset numeeriset menetelmät mustien aukkojen binäärijärjestelmän ymmärtämiseen ja simulointiin. Tätä tarkoitusta varten johdanto yhdistää sekä perustuloksia että joitakin syvällisempiä tuloksia laajoilta fysiikan osa-alueilta kuten suhteellisuusteoriasta ja säteilyhydrodynamiikasta. Johdannon sisältämän materiaalin avulla väitöskirjan julkaisut, ja niiden esittämät tulokset, ovat hyvin ymmärrettävissä. Väitöskirjan julkaisuista ensimmäinen esittelee uusia OJ287-systeemistä saatuja havaintopisteitä, jotka on paikallistettu Harvardin yliopiston observatorion arkiston valokuvauslevyiltä. OJ287:n vuonna 1900 tapahtunut purkaus nähdään ensimmäistä kertaa näissä havaintopisteissä. Uudet havaintopisteet mahdollistivat myös vuoden 1913 purkauksen alun ajoittamisen tarkemmin kuin aiemmin oli mahdollista. Havaitut purkaukset mallinnettiin onnistuneesti simuloimalla OJ287-järjestelmän mustien aukkojen paria ja kertymäkiekkoa. Julkaisussa II käsitellään mekanismeja OJ287:n sekundäärisen mustan aukon spinin kasvamiseen vuorovaikutuksessa primäärin kertymäkiekon ja systeemin magneettikenttien kanssa. Julkaisussa arvioidaan maksimispinin saavuttamisen ja spinin suunnan vakiintumisen aikaskaalat kummallakin mekanismilla. Tutkimuksessa havaitaan sekundäärin spinin olevan todennäköisesti suuri. Julkaisu III esittelee OJ287-systeemissä maaliskuussa 2013 tapahtuneen purkauksen. Purkauksen havaittiin muistuttavan vuosina 1993 ja 2004 tapahtuneita purkauksia, joita kutsutaan yhteisnimityksellä prekursoripurkaus (precursor outburst). Julkaisussa esitellään purkauksen synnylle mekanismi, jossa OJ287-systeemin sekundäärinen musta aukko osuu primäärisen mustan aukon kertymäkiekon koronassa olevaan kaasupilveen. Mekanismin avulla johdetaan arviot prekursoripurkausten kirkkaudelle ja aikaskaalalle. Julkaisussa johdetaan myös ennuste seuraavan prekursoripurkauksen ajankohdalle. Julkaisussa IV käytetään vuosina 2004–2006 kerättyjä havaintoja OJ287- systeemistä lyhyiden jaksollisuuksien etsintään. Julkaisussa varmennetaan systeemissä esiintyvä n. 50 päivän kvasiperiodisuus. Lisäksi tilastollisesti merkittävät 250 päivän ja 3,5 päivän jaksollisuudet havaitaan. Julkaisussa esitetään malli, jossa primäärisen mustan aukon kertymäkiekossa oleva spiraalitiheysaalto aiheuttaa 50 päivän jaksollisuuden. Mallista tehty numeerinen simulaatio tukee tulosta. Systeemin relativistisen suihkun emittoima aikadilatoitunut säteily esitetään aiheuttajaksi 3,5 päivän jaksollisuusaikaskaalalle. Julkaisussa V sovitetaan kierresuihkumalli OJ287-systeemistä tehtyihin optisiin havaintoihin ja millimetri- sekä senttimetriaallonpituuden radiohavaintoihin. Suihkun rakenteen havaitaan olevan kaksijakoinen ja koostuvan ytimestä ja kuoresta. Suihkun kuorella on merkittävästi pienempi Lorentzin gamma-tekijä kuin suihkun ytimellä. Kuoren avautumiskulma ja Lorentztekijä sekä suihkun kierteen aallonpituus raportoidaan julkaisussa ensimmäistä kertaa.

Preparation of mitotic chromosomes of leaf-cutting ants from the genera Atta and Acromyrmex

Some modifications were made to the methodology of Imai et al. (Jpn. J. Genet. 63: 159-185, 1988) for cytogenetic analysis of the leaf-cutting ants Atta sexdens piriventris and Acromyrmex heyeri (Hymenoptera, Formicidae), shortening preparation time and improving chromosomal preparations. The brain ganglia of prepupae were dissected in a 0.0025% hypotonic solution of colchicine, placed on a glass slide on a cold plate (4 ± 1oC) for 20 min. The material was fixed directly on the cold slide (with cold fixative I), macerated with a histological needle and fixed again with fixative I, followed by fixatives II and III, all of them cold. The slide was flame-dried right after the use of fixative III, and it was allowed to air-dry at room temperature for 2 h. The resulting metaphases presented less contracted chromosomes, with separated and well defined sister chromatids at a high frequency, when the material was processed in the manner described and stained with 3% Giemsa in phosphate buffer (pH 6.8) for 15 min.

Endothelium-dependent vasodilation in response to Pseudomonas aeruginosa lipopolysaccharide: an in vitro study on canine arteries

Early systemic arterial hypotension is a common clinical feature of Pseudomonas septicemia. To determine if Pseudomonas aeruginosa endotoxin induces the release of endothelium-derived nitric oxide (EDNO), an endogenous nitrovasodilator, segments of canine femoral, renal, hepatic, superior mesenteric, and left circumflex coronary arteries were suspended in organ chambers (physiological salt solution, 95% O2/5% CO2, pH 7.4, 37oC) to measure isometric force. In arterial segments contracted with 2 µM prostaglandin F2a, Pseudomonas endotoxin (lipopolysaccharide (LPS) serotype 10(Habs) from Pseudomonas aeruginosa (0.05 to 0.50 mg/ml)) induced concentration-dependent relaxation of segments with endothelium (P<0.05) but no significant change in tension of arteries without endothelium. Endothelium-dependent relaxation in response to Pseudomonas LPS occurred in the presence of 1 µM indomethacin, but could be blocked in the coronary artery with 10 µM NG-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of nitric oxide synthesis from L-arginine. The inhibitory effect of L-NMMA on LPS-mediated vasorelaxation of the coronary artery could be reversed by exogenous 100 µM L-arginine but not by 100 µM D-arginine. These experiments indicate that Pseudomonas endotoxin induces synthesis of nitric oxide from L-arginine by the vascular endothelium. LPS-mediated production of EDNO by the endothelium, possibly through the action of constitutive nitric oxide synthase (NOSc), may decrease systemic vascular resistance and may be the mechanism of early hypotension characteristic of Pseudomonas septicemia.

The mechanism of gentisic acid-induced relaxation of the guinea pig isolated trachea: the role of potassium channels and vasoactive intestinal peptide receptors

We examined some of the mechanisms by which the aspirin metabolite and the naturally occurring metabolite gentisic acid induced relaxation of the guinea pig trachea in vitro. In preparations with or without epithelium and contracted by histamine, gentisic acid caused concentration-dependent and reproducible relaxation, with mean EC50 values of 18 µM and Emax of 100% (N = 10) or 20 µM and Emax of 92% (N = 10), respectively. The relaxation caused by gentisic acid was of slow onset in comparison to that caused by norepinephrine, theophylline or vasoactive intestinal peptide (VIP). The relative rank order of potency was: salbutamol 7.9 > VIP 7.0 > gentisic acid 4.7 > theophylline 3.7. Gentisic acid-induced relaxation was markedly reduced (24 ± 7.0, 43 ± 3.9 and 78 ± 5.6%) in preparations with elevated potassium concentration in the medium (20, 40 or 80 mM, respectively). Tetraethylammonium (100 µM), a nonselective blocker of the potassium channels, partially inhibited the relaxation response to gentisic acid, while 4-AP (10 µM), a blocker of the voltage potassium channel, inhibited gentisic acid-induced relaxation by 41 ± 12%. Glibenclamide (1 or 3 µM), at a concentration which markedly inhibited the relaxation induced by the opener of ATP-sensitive K+ channels, levcromakalim, had no effect on the relaxation induced by gentisic acid. Charybdotoxin (0.1 or 0.3 µM), a selective blocker of the large-conductance Ca2+-activated K+ channels, caused rightward shifts (6- and 7-fold) of the gentisic acid concentration-relaxation curve. L-N G-nitroarginine (100 µM), a NO synthase inhibitor, had no effect on the relaxant effect of gentisic acid, and caused a slight displacement to the right in the relaxant effect of the gentisic acid curve at 300 µM, while methylene blue (10 or 30 µM) or ODQ (1 µM), the inhibitors of soluble guanylate cyclase, all failed to affect gentisic acid-induced relaxation. D-P-Cl-Phe6,Leu17[VIP] (0.1 µM), a VIP receptor antagonist, significantly inhibited (37 ± 7%) relaxation induced by gentisic acid, whereas CGRP (8-37) (0.1 µM), a CGRP antagonist, only slightly enhanced the action of gentisic acid. Taken together, these results provide functional evidence for the direct activation of voltage and large-conductance Ca+2-activated K+ channels, or indirect modulation of potassium channels induced by VIP receptors and accounts for the predominant relaxation response caused by gentisic acid in the guinea pig trachea.

Lack of evidence for superantigen activity of Toxoplasma gondii towards human T cells

Toxoplasma gondii is an obligatory intracellular parasite whose life cycle may include man as an intermediate host. More than 500 million people are infected with this parasite worldwide. It has been previously reported that T. gondii contains a superantigen activity. The purpose of the present study was to determine if the putative superantigen activity of T. gondii would manifest towards human T cells. Peripheral blood mononuclear cells (PBMC) from individuals with no previous contact with the parasite were evaluated for proliferation as well as specific Vß expansion after exposure to Toxoplasma antigens. Likewise, PBMC from individuals with the congenital infection were evaluated for putative Vß family deletions in their T cell repertoire. We also evaluated, over a period of one year, the PBMC proliferation pattern in response to Toxoplasma antigens in patients with recently acquired infection. Some degree of proliferation in response to T. gondii was observed in the PBMC from individuals never exposed to the parasite, accompanied by specific Vß expansion, suggesting a superantigen effect. However, we found no specific deletion of Vß (or Valpha) families in the blood of congenitally infected individuals. Furthermore, PBMC from recently infected individuals followed up over a period of one year did not present a reduction of the Vß families that were originally expanded in response to the parasite antigens. Taken together, our data suggest that T. gondii does not have a strong superantigen activity on human T cells.

Ionic radiocontrast inhibits endothelium-dependent vasodilation of the canine renal artery in vitro: possible mechanism of renal failure following contrast medium infusion

To determine if radiocontrast impairs vascular relaxation of the renal artery, segments (4-5 mm in length) of canine renal artery were suspended in vitro in organ chambers to measure isometric force (95% O2/5% CO2, at 37ºC). Arterial segments with and without endothelium were placed at the optimal point of their length-tension relation and incubated with 10 µM indomethacin to prevent synthesis of endogenous prostanoids. The presence of nonionic radiocontrast (iohexol, Omnipaque 350, 1 ml in 25 ml control solution, 4% (v/v)) did not alter endothelium-dependent relaxation to acetylcholine in rings precontracted with both norepinephrine and prostaglandin F2alpha (N = 6). When the rings were precontracted with prostaglandin F2alpha, the presence of ionic contrast did not inhibit the relaxation of the arteries. However, in canine renal arteries contracted with norepinephrine, the presence of ionic radiocontrast (diatrizoate meglumine and diatrizoate sodium, MD-76, 1 ml in 25 ml control solution, 4% (v/v)) inhibited relaxation in response to acetylcholine, sodium nitroprusside (N = 6 in each group), and isoproterenol (N = 5; P < 0.05). Rings were relaxed less than 50% of norepinephrine contraction. Following removal of the contrast, vascular relaxation in response to the agonists returned to normal. These results indicate that ionic radiocontrast nonspecifically inhibits vasodilation (both cAMP-mediated and cGMP-mediated) of canine renal arteries contracted with norepinephrine. This reversible impairment of vasodilation could inhibit normal renal perfusion and act as a mechanism of renal failure following radiocontrast infusion. In the adopted experimental protocol the isoproterenol-induced relaxation of renal arteries precontracted with norepinephrine was more affected, suggesting a pivotal role of the cAMP system.

Cardiac and vascular effects of diltiazem, dobutamine and amrinone, drugs used after myocardial revascularization

Hemodynamic care during postoperative management of myocardial revascularization should include vasorelaxing drugs to insure adequate graft and coronary flow, and stimulation of stroke volume to maintain vascular perfusion pressure. We tested the cardiac (inotropic and lusitropic) and vascular (relaxant) effects of diltiazem (0.1 nM to 0.1 mM), dobutamine (10 µM to 10 mM) and amrinone (10 µM to 1 mM) on isolated rat atria and thoracic aorta, and also on isolated human saphenous vein (HSV) and human mammary artery (HMA). Dobutamine produced a maximal positive inotropic effect (+dF/dt max = 29 ± 7%) at its ED50 for aortic relaxation (88 ± 7 µM). Conversely, at their ED50 for aortic relaxation diltiazem depressed myocardial contractility and amrinone did not exhibit myocardial effects. In HSV and HMA contracted with 80 mM potassium, diltiazem and dobutamine (but not amrinone) had a vasorelaxant activity similar to that in rat aorta. Norepinephrine-contracted human vessels were significantly more sensitive than potassium-contracted vessels to the relaxant effect of amrinone (ED50 HMA = 15 ± 5 µM, ED50 HSV = 72 ± 31 µM, P < 0.05). We conclude that at concentrations still devoid of myocardial effects dobutamine and amrinone are effective dilators in graft segment vessels and rat aorta contracted by membrane depolarization. If the difference between aortic and myocardial tissue still holds in human tissues, at the appropriate concentrations these drugs should be expected to improve cardiac performance while still contributing to the maintenance of graft patency.

Protective effect of N-acetylcysteine against oxygen radical-mediated coronary artery injury

The present study investigated the protective effect of N-acetylcysteine (NAC) against oxygen radical-mediated coronary artery injury. Vascular contraction and relaxation were determined in canine coronary arteries immersed in Kreb's solution (95% O2-5% CO2), incubated or not with NAC (10 mM), and exposed to free radicals (FR) generated by xanthine oxidase (100 mU/ml) plus xanthine (0.1 mM). Rings not exposed to FR or NAC were used as controls. The arteries were contracted with 2.5 µM prostaglandin F2alpha. Subsequently, concentration-response curves for acetylcholine, calcium ionophore and sodium fluoride were obtained in the presence of 20 µM indomethacin. Concentration-response curves for bradykinin, calcium ionophore, sodium nitroprusside, and pinacidil were obtained in the presence of indomethacin plus Nomega-nitro-L-arginine (0.2 mM). The oxidative stress reduced the vascular contraction of arteries not exposed to NAC (3.93 ± 3.42 g), compared to control (8.56 ± 3.16 g) and to NAC group (9.07 ± 4.0 g). Additionally, in arteries not exposed to NAC the endothelium-dependent nitric oxide (NO)-dependent relaxation promoted by acetylcholine (1 nM to 10 µM) was also reduced (maximal relaxation of 52.1 ± 43.2%), compared to control (100%) and NAC group (97.0 ± 4.3%), as well as the NO/cyclooxygenase-independent receptor-dependent relaxation provoked by bradykinin (1 nM to 10 µM; maximal relaxation of 20.0 ± 21.2%), compared to control (100%) and NAC group (70.8 ± 20.0%). The endothelium-independent relaxation elicited by sodium nitroprusside (1 nM to 1 µM) and pinacidil (1 nM to 10 µM) was not affected. In conclusion, the vascular dysfunction caused by the oxidative stress, expressed as reduction of the endothelium-dependent relaxation and of the vascular smooth muscle contraction, was prevented by NAC.

Calcium antagonism and the vasorelaxation of the rat aorta induced by rotundifolone

The vasorelaxing activity of rotundifolone (ROT), a major constituent (63.5%) of the essential oil of Mentha x villosa, was tested in male Wistar rats (300-350 g). In isolated rat aortic rings, increasing ROT concentrations (0.3, 1, 10, 100, 300, and 500 µg/ml) inhibited the contractile effects of 1 µM phenylephrine and of 80 or 30 mM KCl (IC50 values, reported as means ± SEM = 184 ± 6, 185 ± 3 and 188 ± 19 µg/ml, N = 6, respectively). In aortic rings pre-contracted with 1 µM phenylephrine, the smooth muscle-relaxant activity of ROT was inhibited by removal of the vascular endothelium (IC50 value = 235 ± 7 µg/ml, N = 6). Furthermore, ROT inhibited (pD2 = 6.04, N = 6) the CaCl2-induced contraction in depolarizing medium in a concentration-dependent manner. In Ca2+-free solution, ROT inhibited 1 µM phenylephrine-induced contraction in a concentration-dependent manner and did not modify the phasic contractile response evoked by caffeine (20 mM). In conclusion, in the present study we have shown that ROT produces an endothelium-independent vasorelaxing effect in the rat aorta. The results further indicated that in the rat aorta ROT is able to induce vasorelaxation, at least in part, by inhibiting both: a) voltage-dependent Ca² channels, and b) intracellular Ca2+ release selectively due to inositol 1,4,5-triphosphate activation. Additional studies are required to elucidate the mechanisms underlying ROT-induced relaxation.

Endothelium-dependent and -independent hepatic artery vasodilatation is not impaired in a canine model of liver ischemia-reperfusion injury

We investigated whether hepatic artery endothelium may be the earliest site of injury consequent to liver ischemia and reperfusion. Twenty-four heartworm-free mongrel dogs of either sex exposed to liver ischemia/reperfusion in vivo were randomized into four experimental groups (N = 6): a) control, sham-operated dogs, b) dogs subjected to 60 min of ischemia, c) dogs subjected to 30 min of ischemia and 60 min of reperfusion, and d) animals subjected to 45 min of ischemia and 120 min of reperfusion. The nitric oxide endothelium-dependent relaxation of hepatic artery rings contracted with prostaglandin F2a and exposed to increasing concentrations of acetylcholine, calcium ionophore A23187, sodium fluoride, phospholipase-C, poly-L-arginine, isoproterenol, and sodium nitroprusside was evaluated in organ-chamber experiments. Lipid peroxidation was estimated by malondialdehyde activity in liver tissue samples and by blood lactic dehydrogenase (LDH), serum aspartate aminotransferase (AST) and serum alanine aminotransferase (ALT) activities. No changes were observed in hepatic artery relaxation for any agonist tested. The group subjected to 45 min of ischemia and 120 min of reperfusion presented marked increases of serum aminotransferases (ALT = 2989 ± 1056 U/L and AST = 1268 ± 371 U/L; P < 0.01), LDH = 2887 ± 1213 IU/L; P < 0.01) and malondialdehyde in liver samples (0.360 ± 0.020 nmol/mgPT; P < 0.05). Under the experimental conditions utilized, no abnormal changes in hepatic arterial vasoreactivity were observed: endothelium-dependent and independent hepatic artery vasodilation were not impaired in this canine model of ischemia/reperfusion injury. In contrast to other vital organs and in the ischemia/reperfusion injury environment, dysfunction of the main artery endothelium is not the first site of reperfusion injury.

A key role for Na+/K+-ATPase in the endothelium-dependent oscillatory activity of mouse small mesenteric arteries

Oscillatory contractile activity is an inherent property of blood vessels. Various cellular mechanisms have been proposed to contribute to oscillatory activity. Mouse small mesenteric arteries display a unique low frequency contractile oscillatory activity (1 cycle every 10-12 min) upon phenylephrine stimulation. Our objective was to identify mechanisms involved in this peculiar oscillatory activity. First-order mesenteric arteries were mounted in tissue baths for isometric force measurement. The oscillatory activity was observed only in vessels with endothelium, but it was not blocked by L-NAME (100 µM) or indomethacin (10 µM), ruling out the participation of nitric oxide and prostacyclin, respectively, in this phenomenon. Oscillatory activity was not observed in vessels contracted with K+ (90 mM) or after stimulation with phenylephrine plus 10 mM K+. Ouabain (1 to 10 µM, an Na+/K+-ATPase inhibitor), but not K+ channel antagonists [tetraethylammonium (100 µM, a nonselective K+ channel blocker), Tram-34 (10 µM, blocker of intermediate conductance K+ channels) or UCL-1684 (0.1 µM, a small conductance K+ channel blocker)], inhibited the oscillatory activity. The contractile activity was also abolished when experiments were performed at 20°C or in K+-free medium. Taken together, these results demonstrate that Na+/K+-ATPase is a potential source of these oscillations. The presence of α-1 and α-2 Na+/K+-ATPase isoforms was confirmed in murine mesenteric arteries by Western blot. Chronic infusion of mice with ouabain did not abolish oscillatory contraction, but up-regulated vascular Na+/K+-ATPase expression and increased blood pressure. Together, these observations suggest that the Na+/K+ pump plays a major role in the oscillatory activity of murine small mesenteric arteries.

Bowslaugh Family fonds, 1860-1861, 1868, 1876-1877, 1880-1882, 1886-1890, 1892-1893, 1895, 1898-1899, n.d.

Edward W. Bowslaugh (1843-1923) was the son of Jacob and Anna (Beamer) Bowslaugh. Edward Bowslaugh married Mary Southward, and the couple had six children, Edgar Morley, Edward Freeman, twins Alfred Malcolm and Alice Mary, Annie Olivia, John Jacob and Mabel Florence. Edward W. Bowslaugh was a farmer, contractor and owner of the Grimsby Planing Mills in Grimsby, Ont. and Bowslaugh’s Planing Mill in Kingsville, Ont. The mills manufactured door and sash trim and other wood related products. Some customers contracted the firm to provide wood products for cottages being built at Grimsby Park, the Methodist camp ground. Some time before 1885 Edward Bowslaugh and his family moved to Kingsville, Ont. to open up a new planing mill and door and sash manufactory. He later sold the Grimsby Planing Mills to Daniel Marsh. The diaries and account books include many names of workers as well as friends and family members residing in the Grimsby and Kingsville areas. James M. Bowslaugh (1841-1882) was the son of Jacob and Anna (Beamer) Bowslaugh. James married first Anna Catharine Merritt and after her death in 1875 he married Mary Gee in 1877. James and Anna had three children, Eliza, James Herbert, George Hiram, all died very young. James and Mary Gee had one son, Charles Leopold Kenneth Frederich Bowslaugh, b. 1881. James Bowslaugh was a farmer and lumberman, much like his younger brother Edward. James’ early diaries often note the activities of himself and his brother Edward. Both Edward and James were heavily involved in the Methodist church, teaching or leading Sunday school and attending prayer meetings. Alfred M. Bowslaugh b. 1873 was the son of Edward W. Bowslaugh and his wife Mary Southward. The school notebook is from his days as a student in Kingsville, Ont.