Myocardial perfusion imaging in the setting of coronary artery stenosis and acute myocardial infarction using venous injection of a second-generation echocardiographic contrast agent

BACKGROUND: We hypothesized that by producing excellent myocardial opacification, venous injection of FS-069 coupled with intermittent harmonic imaging (IHI) can be used to determine the presence and severity of coronary stenoses during hyperemia, the size of the risk area during coronary occlusion, and the extent of myocardial salvage after reperfusion. METHODS AND RESULTS: Twelve dogs were imaged both continuously and intermittently (every end systole) in the fundamental (2 MHz) and harmonic (transmit at 2 and receive at 4 MHz) modes. FS-069 (1 mL) was injected intravenously for all stages and modes of imaging. Myocardial video intensity was severalfold (P<.01) higher during IHI than all other modes of imaging. Perfusion defects were difficult to measure during continuous and intermittent fundamental imaging and during continuous harmonic imaging. In comparison, the defects were clearly demarcated during IHI. When this mode was used, the magnitude of perfusion mismatch during hyperemia in the presence of a coronary stenosis correlated closely with the magnitude of flow mismatch when radiolabeled microspheres were used (r=.94). The perfusion defect sizes during coronary occlusion and reperfusion also correlated closely with postmortem risk area (r=.89) and infarct size (r=.96), respectively. CONCLUSIONS: Venous injection of FS-069 coupled with IHI produces excellent myocardial opacification. This approach can be used to determine the severity of coronary stenoses during hyperemia, the size of the risk area during coronary occlusion, and the extent of myocardial salvage after reperfusion. This approach, therefore, holds promise in the clinical setting.     

Phosphatase-negative mutants of Legionella pneumophila and their behavior in mammalian cell infection

Microbial phosphatases are known or suspected to play a role in the pathogenesis of several intracellular pathogens, including Legionella micdadei. Legionella pneumophila also possess phosphatase activities, but their possible roles in cellular infection are unknown. We generated mutants of a serogroup 1 isolate of L. pneumophila that lack the major phosphatase. Isolation of a Pho- mutant after random mutagenesis with transposon MudII4041 allowed us to dissociate the major alkaline phosphatase (pH optimum approximately 8) from a minor acid phosphatase activity. Both activities were concentrated in the bacterial periplasm. The gene encoding the major alkaline phosphatase (pho) was cloned by expression in E. coli and used to generate a site directed mutation in two L. pneumophila strains. Each parent-mutant pair was compared in a U937 cell tissue culture assay for capacity to infect, lyse, and grow within mammalian cells. Although the parental stains differed in their U937 cell cytopathicity, neither was significantly more infective than its Pho- derivative, suggesting that the alkaline phosphatase activity is not essential for cellular infection. Because they are not attenuated, Pho- mutants can be used to generate gene fusions with E. coli alkaline phosphatase to study and secretion and cellular infectivity in L. pneumophila.     

The expression of voltage-dependent calcium channel beta subunits in human cerebellum

The beta subunits of voltage-dependent calcium channels, exert marked regulatory effects on the biophysical and pharmacological properties of this diverse group of ion channels. However, little is known about the comparative neuronal expression of the four classes of beta genes in the CNS. In the current investigation we have closely mapped the distribution of beta1, beta2, beta3 and beta4 subunits in the human cerebellum by both in situ messenger RNA hybridization and protein immunohistochemistry. To our knowledge, these studies represent the first experiments in any species in which the detailed localization of each beta protein has been comparatively mapped in a neuroanatomically-based investigation. The data indicate that all four classes of beta subunits are found in the cerebellum and suggest that in certain neuronal populations they may each be expressed within the same cell. Novel immunohistochemical results further exemplify that the beta voltage-dependent calcium channel subunits are regionally distributed in a highly specific manner and studies of Purkinje cells indicate that this may occur at the subcellular level. Preliminary indication of the subunit composition of certain native voltage-dependent calcium channels is suggested by the observation that the distribution of the beta3 subunit in the cerebellar cortex is identical to that of alpha(1E). Our cumulative data are consistent with the emerging view that different native alpha1/beta subunit associations occur in the CNS.     

Cells of the perireticular nucleus project to the developing neocortex of the rat

The perireticular nucleus is a recently described thin sheet of small cells among the fibres of the internal capsule, lying lateral to the thalamic reticular nucleus and medial to the globus pallidus (Clemence and Mitrofanis [1992]. J. Comp. Neurol. 322:167-180). During development, the perireticular nucleus is relatively large, lying in the path of the growing corticofugal and thalamocortical axons and filling the area of the internal capsule lateral to the thalamic reticular nucleus. After these axons have formed their connections, the perireticular nucleus rapidly decreases in size, leaving only a few cells in the adult (Mitrofanis [1992] J. Comp. Neurol. 320:161-181). In this study, we aimed to investigate the connections between the developing cortex and thalamus by making injections of tracer into the cortical plate. Injections of Horse Radish Peroxidase (HRP), Wheat Germ Agglutinin bound to HRP (WGA-HRP) and 1'dioctadecyl-3,3,3',3 tetramethycarbocyanine perchlorate (DiI) were made in vivo between embryonic day (E) 18 and adult and DiI was placed in the fixed brains of rats aged between E16 and postnatal day (P)1. Between E17 and P10, the retrograde perikaryal labelling resulting from these injections revealed a transient projection from the perireticular nucleus to the ipsilateral cortical plate. No cells were labelled in the thalamic reticular nucleus. This suggests that the perireticular nucleus must be regarded as a group of cells distinct from the thalamic reticular nucleus and having a separate role in development. Comparisons between the perireticular cells and the cells of the cortical subplate suggest that both may be playing comparable roles in early development, possibly guiding fibres towards their end stations or serving to rearrange the complex mapped projections linking the thalamus and cortex.     

Endocrine and reproductive function in ewes exposed to the organochlorine pesticides lindane or pentachlorophenol

The effects of lindane (LIN, gamma-hexachlorocyclohexane) and pentachlorophenol (PCP) on reproduction and general endocrine function were examined in breeding ewes as a model for wild and domestic ungulates, which may be exposed to low levels of pesticides that are potential endocrine-disrupting chemicals. Ewes (n = 13/group) were fed either a control untreated diet (CON), or a diet treated with LIN (1 mg/kg/d) or PCP (1 mg/kg/d) during the 5 wk prior to mating and throughout pregnancy and lactation. Mating response, ovulation rate, follicle and corpus luteum size, gestation length, pregnancy rate, lambing rate, and lamb birth weight were recorded. After weaning, 6 ewes from each group were bled at 15-min intervals for 8 h during the day and night and for 1 h before and 5 h after i.v. administration of gonadotropin-releasing hormone, thyroid-stimulating hormone (TSH), and adrenocorticotropin, to measure serum concentrations of luteinizing hormone, follicle-stimulating hormone, thyroxine (T4), and cortisol. Ewes were then killed and endocrine tissues examined histologically. Pregnancy rate as a result of matings taking place at the synchronized estrus was significantly decreased by the lindane treatment However, PCP and lindane did not markedly affect any other aspect of reproductive function studied. In PCP-treated ewes, serum concentrations of T4 were significantly reduced compared to control ewes during the day and night; however, the T4 response to TSH was not altered by PCP treatment. No other measured endocrine parameters were consistently affected by lindane or PCP. Thyroid follicle size was significantly increased in the LIN and PCP ewes compared to the control ewes. Low serum concentrations of T4 in the PCP ewes may have resulted in increased TSH secretion and increased thyroid follicle size. In conclusion, although pesticide treatments had no serious adverse effects on reproductive function in breeding ewes, PCP reduced T4 concentration, which in the long term could influence reproductive and general performance.     

Endomorphin-1 and endomorphin-2 activate mu-opioid receptors in myenteric neurons of the guinea-pig small intestine

An association between hyperhomocysteinemia and premature atherosclerosis in patients with non-insulin-dependent diabetes mellitus (NIDDM) has recently been described. Little is known about the role of insulin in homocysteine [H(e)] metabolism. We measured plasma H(e) concentrations in the fasting state and during a hyperinsulinemic-euglycemic clamp in normal subjects and patients with NIDDM. Plasma H(e) decreased significantly from 7.2 +/- 2.6 to 6.0 +/- 2.7 mmol/L (P < .01) in normal subjects, but did not change in patients with NIDDM (6.0 +/- 2.7 to 5.9 +/- 2.5 mmol/L, respectively). These data suggest that plasma H(e) concentrations are regulated by acute hyperinsulinemia in normal subjects, but not in insulin-resistant NIDDM subjects. These abnormalities may have implications for the pathogenesis of premature vascular disease associated with NIDDM.