Septicaemia and arthritis in pigs experimentally infected with Pasteurella multocida capsular serotype A.

Twenty-five caesarean-derived, colostrum-deprived (CDCD) pigs and 18 specific pathogen-free pigs, aged 8 to 14 weeks, were inoculated intranasally or intratracheally with Pasteurella multocida capsular serotype A, isolated from a severe pneumonic lesion in a growing pig. The pigs were killed for necropsy on day 6 or 14 post-inoculation (PI) or, in the case of the only fatally infected animal, examined at the time of death. One CDCD pig, inoculated intratracheally with 5 ml of a bacterial suspension containing 1.7x10(9) colony-forming-units/ml, died of septicaemia on day 1 PI. Histological lesions such as severe pleuropneumonia, thrombi in glomerular capillaries, haemorrhage of the spleen, and abscesses in the tonsillar crypts were observed. The organism was recovered from a number of sites and its antigens were detected immunohistochemically in the pneumonic lesions, blood vessels of the tissues, and tonsillar crypts in the dead pig. Pneumonia, pleural adhesions and suppurative arthritis in the extremital joints were observed grossly in 3/29, 8/29 and 7/29 intratracheally inoculated pigs, respectively. In intranasally inoculated pigs, no macroscopical abnormalities were seen; histologically, however, exudative bronchopneumonia and fibrinous pleurisy were observed in 9/14 and 4/14 pigs, respectively. No significant changes were seen in the tissues of uninfected control pigs. The organism was recovered from the lesions and P. multocida type A antigen was demonstrated immunohistochemically. The organism was rarely recovered from the liver, spleen or lymph nodes (bronchopulmonary or mesenteric). The results suggest that P. multocida capsular serotype A alone can cause not only pneumonia in pigs but also septicaemia or arthritis.

Effect of non-aqueous phase liquid on biodegradation of PAHs in spilled oil on tidal flat.

Biodegradation rates of polycyclic aromatic hydrocarbons (PAHs) in spilled oil stranded on tidal flats were studied using model reactors to clarify the effects of NAPL on the biodegradation of PAHs in stranded oil on tidal flat with special emphasis on the relationship between dissolution rates of PAHs into water and viscosity of NAPL. Biodegradation of PAHs in NAPL was limited by the dissolution rates of PAHs into water. Biodegradation rate of chrysene was smaller than that for acenaphthene and phenanthrene due to the smaller dissolution rates. Dissolution rates of PAHs in fuel oil C were smaller those in crude oil due to high viscosity of fuel oil C. Therefore, biodegradation rates of PAHs in fuel oil C were smaller than those in crude oil. Biodegradation rates of PAHs in NAPL with slow decrease rate like fuel oil C were slower than those in NAPL with rapid decrease like crude oil. The smaller decrease rate of fuel oil C than crude oil was due to higher viscosity of fuel oil C. Therefore, not only the dissolution rate of PAHs but also the decrease rates of NAPL were important factors for the biodegradation of PAHs.

Attention and visual interference stimulation affect somatosensory processing: a magnetoencephalographic study.

In our previous study, continuous visual (cartoon and random dot motion) and auditory (music) stimulation changed the somatosensory evoked magnetic fields following electrical stimulation of the median nerve in human subjects. They enhanced the middle-latency components (3M and 4M) generated in the contralateral primary somatosensory cortex, and reduced the MI component generated in the ipsilateral secondary somatosensory cortex. We speculated that such interference effects were caused by activation of polymodal neurons in areas 5 and/or 7 of the parietal lobe as well as in the medial superior temporal region and superior temporal sulcus. However, we could not exclude the effect of attention on such interference effects. In the present study, to know the effect of attention on visual and auditory interference in these changes, we stimulated the bilateral median nerves unilaterally in a random order, and asked subjects to count the number of times the left median nerve was stimulated while visual or auditory interference was applied. Five components (1M-5M) were identified in the hemisphere contralateral to the stimulated nerve and only one component (MI) was found in the ipsilateral hemisphere. The 3M and 4M components (33-75 ms in latency) were enhanced by both attention and visual interference stimulation but not by auditory stimulation. The 5M component (70-115 ms) and MI component (70-133 ms) were enhanced by attention, but were not changed by attention together with visual or auditory interference. Summarizing the results of our previous study and the present study, (1) visual interference alone enhanced the 3M and 4M but reduced the MI, and (2) attention alone also enhanced the 3M and 4M, and enhanced the 5M and MI. As a result, (3) visual interference with attention enhanced the 3M and 4M more, and showed no significant change of the 5M and MI. This was compatible with a summation of the effects caused by visual interference alone and attention alone, but some interactions between visual interference and attention might have taken place. The locations of dipoles of all components were not significantly changed by attention or any interference stimulation. These findings support the idea that there are significant interactions of activities relating to somatosensory stimulation, visual stimulation and cognitive function, in both the primary and the secondary somatosensory cortex in humans.

(99m)Tc-HYNIC-derivatized ternary ligand complexes for (99m)Tc-labeled polypeptides with low in vivo protein binding.

6-Hydrazinopyridine-3-carboxylic acid (HYNIC) is a representative agent used to prepare technetium-99m ((99m)Tc)-labeled polypeptides with tricine as a coligand. However, (99m)Tc-HYNIC-labeled polypeptides show delayed elimination rates of the radioactivity not only from the blood but also from nontarget tissues such as the liver and kidney. In this study, a preformed chelate of tetrafluorophenol (TFP) active ester of [(99m)Tc](HYNIC)(tricine)(benzoylpyridine: BP) ternary complex was synthesized to prepare (99m)Tc-labeled polypeptides with higher stability against exchange reactions with proteins in plasma and lysosomes using the Fab fragment of a monoclonal antibody and galactosyl-neoglycoalbumin (NGA) as model polypeptides. When incubated in plasma, [(99m)Tc](HYNIC-Fab)(tricine)(BP) showed significant reduction of the radioactivity in high molecular weight fractions compared with [(99m)Tc](HYNIC-Fab)(tricine)(2.) When injected into mice, [(99m)Tc](HYNIC-NGA)(tricine)(BP) was metabolized to [(99m)Tc](HYNIC-lysine)(tricine)(BP) in the liver with no radioactivity detected in protein-bound fractions in contrast to the observations with [(99m)Tc](HYNIC-NGA)(tricine)(2.) In addition, [(99m)Tc](HYNIC-NGA)(tricine)(BP) showed significantly faster elimination rates of the radioactivity from the liver as compared with [(99m)Tc](HYNIC-NGA)(tricine)(2.) Similar results were observed with (99m)Tc-labeled Fab fragments where [(99m)Tc](HYNIC-Fab)(tricine)(BP) exhibited significantly faster elimination rates of the radioactivity not only from the blood but also from the kidney. These findings indicated that conjugation of [(99m)Tc](HYNIC)(tricine)(BP) ternary ligand complex to polypeptides accelerated elimination rates of the radioactivity from the blood and nontarget tissues due to low binding of the [(99m)Tc](HYNIC)(tricine)(BP) complex with proteins in the blood and in the lysosomes. Such characteristics would render the TFP active ester of [(99m)Tc](HYNIC)(tricine)(BP) complex attractive as a radiolabeling reagent for targeted imaging.

Comparative localization of Dax-1 and Ad4BP/SF-1 during development of the hypothalamic-pituitary-gonadal axis suggests their closely related and distinct functions.

Two nuclear receptors, Ad4BP/SF-1 and Dax-1, are essential regulators for development and function of the mammalian reproductive system. Similarity in expression sites, such as adrenal glands, gonads, pituitary, and hypothalamus, suggests a functional interaction, and the phenotype similarities were manifested in Ad4BP/SF-1-deficient mice and in cases of natural human mutations of Dax-1. In this study, quantitative reverse transcriptase polymerase chain reaction analyses revealed that expression profiles of Dax-1 in embryonic gonads are different between the two sexes and also from those of Ad4BP/SF-1. Immunohistochemical analyses clarified the spatial and temporal expressions of the Dax-1 protein during development of tissues composing the hypothalamic-pituitary-gonadal axis. During gonadal development, Dax-1 occurred after Ad4BP/SF-1 exhibiting a sexually dimorphic expression pattern at indifferent stages, indicating a possibility of Dax-1 involvement in earliest sex differentiation. When cord formation begins in the testis at embryonic day 12.5 (E12.5), Dax-1 was expressed strongly in Sertoli cells, but its expression level markedly decreased in Sertoli cells and increased in interstitial cells between E13.5 and E17.5. In the female, Dax-1 was strongly expressed in the entire ovarian primordium from E12.5 until E14.5, and then its expression level was decreased and limited to cells near the surface epithelium between E17.5 and postnatal day 0 (P0). During postnatal development of the testis, the variable staining of Dax-1 in Sertoli cells was detected as early as P7 and Dax-1-expressing Leydig cells became rare. In the postnatal ovary, Dax-1 expression was detected in granulosa cells with variable staining intensity, and occasionally in interstitial cells. During pituitary organogenesis, Dax-1 but not Ad4BP/SF-1 was expressed in the dorsal part of Rathke's pouch from E9.5. Later in development after E14.5, the distribution of Dax-1 overlapped with that of Ad4BP/SF-1, being restricted to gonadotropic cells in the anterior pituitary. In the ventromedial hypothalamus (VMH), Dax-1 and Ad4BP/SF-1 were mostly colocalized throughout the embryonic and postnatal development. Thus, the coexpression of Dax-1 and Ad4BP/SF-1 indicates their closely related functions in the development of the reproductive system. Furthermore, we noticed the presence of cells that express Dax-1 but not Ad4BP/SF-1, further indicating additional functions of Dax-1 in an Ad4BP/SF-1-independent molecular mechanism.

[Effect of dynamic cardiomyoplasty on the left ventricular function and hemodynamics in chronic canine models].

This study was undertaken to examine the effect of cardiac assist and left ventricular function after dynamic cardiomyoplasty (DCMP). In the first group (GI) of 10 mongrel dogs DCMP was drived immediately after wrapping both ventricles by latissmus dorsi muscle flap (LDMF). In the second group (GII) of 10 mongrel dogs DCMP was derived over 6 weeks after production of DCMP for achievement of complete adhesion between LDMF and myocardium. In the both groups, aortic pressure, cardiac output, left ventricular systolic pressure, and ejection fraction of the left ventricle were significantly increased by DCMP driving (p < 0.001). But left ventricular systolic pressure was remarkably increased in GII compared with that of GI (21.2 +/- 10.2% versus 14.0 +/- 9.6%, p < 0.001), and end diastolic pressure of the left ventricle was apparently decreased in GII (61.6 +/- 42.3% p < 0.05). Thus, satisfactory results were recognized that cardiac assist for left ventricular function was enhanced after completion of adhesion between myocardium and LDMF. Echocardiography in GII demonstrated that left ventricular systolic dimention was significantly decreased from 33.8 +/- 1.0 mm to 27.6 +/- 1.2 mm (p < 0.001). Thus, left ventricular fractional shortening was significantly increased from 24.0 +/- 2.4% to 38.0 +/- 2.6% (p < 0.001). However, left ventricle end-diastolic dimention was not changed even during DCMP driving. So disturbance in left ventricular function during diastole could not be recognized. In conclusion, especially after adhesion of both muscles of LDMF and myocardium, effect of cardiac assist was remarkably enhanced, and disturbance of diastolic function of the left ventricle could not be observed.

[Effect of dynamic cardiomyoplasty on coronary arterial blood flow].

We investigated whether or not dynamic cardiomyoplasty adversely affected coronary arterial blood flow (CABF) through compression of the coronary arteries by muscular contraction during systole and incomplete relaxation of the skeletal muscle flap during diastole. Dynamic cardiomyoplasty was performed in 20 mongrel dogs using a left latissimus dorsi muscle flap, paced synchronously with the R wave of the electrocardiogram. A Doppler catheter (3 F in diameter) was placed in the left main trunk of the coronary artery to analyze the instantaneous changes of coronary arterial blood flow velocity by fast Fourier transformation analysis. We compared both systolic and diastolic properties during assisted versus unassisted cardiac cycles by calculating the peak velocity and the time velocity integrate (TVI). A significant enhancement of systolic CABF was recognized by increases in the systolic peak velocity (26.5 +/- 29.2%) and TVI (20.2 +/- 38.6%). The improved systolic CABF was consistent with an increase in systolic aortic pressure (15.5 +/- 4.3%) and stroke volume (42.8 +/- 11.2%). CABF was also enhanced in diastole because a significant increase of diastolic peak velocity (4.4 +/- 9.4%) and TVI (11.0 +/- 16.7%) was observed. Enhancement of diastolic CABF was associated with the augmentation of cardiac function and the reduction of left ventricular end-diastolic pressure. It could be concluded that CABF was increased by the enhancement of cardiac function as a result of dynamic cardiomyoplasty leading to an increase of cardiac output and aortic pressure and a decrease of left ventricular end-diastolic pressure.