A semiquantitative analysis of reactive astrogliosis demonstrates its correlation with the number of intact motor neurons after transient spinal cord ischemia

ObjectiveWe evaluated the relationship between reactive astrogliosis and delayed motor neuron death after transient spinal cord ischemia in rabbits using a semiquantitative analysis of glial fibrillary acidic protein expression.MethodsSpinal cord ischemia was induced by means of balloon occlusion of the infrarenal aorta for 15 minutes at 39°C in 18 New Zealand white rabbits. At 1, 3, and 7 days after reperfusion, 6 animals at each time point were killed, and the spinal cord was removed for histologic and immunohistochemical study. The variables analyzed were (1) neurologic function (Johnson score) at every 24 hours after reperfusion, (2) the number of intact motor neurons and terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate-biotin nick-end labeling–positive positive neurons, and (3) expression of glial fibrillary acidic protein in the gray and white matter, which was expressed as the percentage of stained area.ResultsAll animals presented delayed motor neuron death. The number of intact neurons decreased correlatively with neurologic function. No obvious terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate-biotin nick-end labeling–positive cells were observed. Glial fibrillary acidic protein expression increased with time in both the gray and white matter, representing the development of reactive astrogliosis. Significant correlation was found between glial fibrillary acidic protein expression and the number of intact motor neurons on the third day in both the gray (r2 = 0.726, P = .031) and white (r2 = 0.927, P = .002) matter.ConclusionsReactive astrogliosis 3 days after transient spinal cord ischemia correlates with the number of intact motor neurons. Our method for semiquantitative analysis of reactive astrogliosis is simple and reproducible and seems useful for such experimental studies

The optimal treatment strategy for secondary mitral regurgitation: a subject of ongoing debate

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Slope in preload recruitable stroke work relationship predicts survival after left ventriculoplasty and mitral repair in patients with idiopathic cardiomyopathy

AbstractBackgroundLeft ventriculoplasty (LVP) and mitral valve plasty (MVP) are sometimes effective for patients with idiopathic dilated cardiomyopathy (DCM) who are not eligible for heart transplantation. Strict patient selection is warranted for these controversial procedures.Methods and resultsThe subjects were 18 patients with idiopathic DCM and mitral regurgitation who had not been indicated for heart transplantation due to either older age or patient refusal, and who underwent LVP and MVP. Their mean age was 57±14 years and 50% were dependent on catecholamine infusion. The preload recruitable stroke work (PRSW) relationship and its slope (Mw) were estimated by a single-beat technique using transthoracic echocardiography. There were one 30-day mortality and six (33%) hospital deaths due to heart failure. The one-year survival rate was 50%. Left ventricular end-diastolic dimension (LVDd) decreased from 77±11 to 68±11mm (p=0.001) whereas the ejection fraction did not change. Preoperative Mw was significantly higher in one-year survivors than that in non-survivors (54±17ergcm−3103 vs. 31±10ergcm−3103, p=0.005). Preoperative LVDd was not different between the groups. The cut-off value of 42ergcm−3103 for Mw predicted one-year survival with high sensitivity (100%) and specificity (77%).ConclusionsMw, the slope in the PRSW relationship, may predict survival after LVP and MVP in patients with idiopathic DCM

Seed-specific expression of truncated OsGAD2 produces GABA-enriched rice grains that influence a decrease in blood pressure in spontaneously hypertensive rats

Gamma-aminobutyric acid (GABA) is a four-carbon amino acid that is commonly present in living organisms and functions as a major inhibitory neurotransmitter in mammals. It is understood to have a potentially anti-hypertensive effect in mammals. GABA is synthesized from glutamate by glutamate decarboxylase (GAD). In plants, GAD is regulated via its calmodulin-binding domain (CaMBD) by Ca2+/CaM. We have previously reported that a C-terminal truncated version of one of the five rice GAD isoforms, GAD2ΔC, revealed higher enzymatic activity in vitro and that its over-expression resulted in exceptionally high GABA accumulation (Akama and Takaiwa, J Exp Bot 58:2699–2607, 2007). In this study, GAD2ΔC, under the control of the rice glutelin promoter (GluB-1), was introduced into rice cells via Agrobacterium-mediated transformation to produce transgenic rice lines. Analysis of the free amino acid content of rice grains revealed up to about a 30-fold higher level of GABA than in non-transformed rice grains. There were also very high levels of various free protein amino acids in the seeds. GABA-enriched rice grains were milled to a fine powder for oral administration to spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto rats (WKYs). Six weeks of administration showed that transgenic rice brings about a 20 mmHg decrease in blood pressure in two different kinds of SHRs, while there was no significant hypotensive effect in WKYs. These results suggest an alternative way to control and/or cure hypertension in humans with GABA-enriched rice as part of a common daily diet

Protective effects of trehalose preconditioning on cardiac and coronary endothelial function through eNOS signaling pathway in a rat model of ischemia-reperfusion injury

Coronary endothelial dysfunction is a major cause of ischemia-reperfusion (I/R) injury. Trehalose, a natural disaccharide, has been reported to ameliorate endothelial dysfunction during aging by activating endothelial nitric oxide synthase (eNOS); however, its role in I/R injury is unknown. This study evaluated the effects of trehalose preconditioning on cardiac and coronary endothelial function after I/R. Langendorff-perfused rat hearts underwent 30 min of global ischemia followed by 80 min of reperfusion with or without trehalose preconditioning. Rate pressure product (RPP) and coronary flow (CF) were measured during reperfusion. Perivascular edema was assessed by hematoxylin and eosin staining. Myocardial oxidative stress and apoptosis were evaluated by immunohistochemistry and TUNEL staining, respectively. eNOS dimerization was determined by western blotting. An eNOS inhibitor was used to examine the role of eNOS. Trehalose preconditioning showed a higher recovery rate after I/R as indicated by high RPP (control vs. trehalose, 28 +/- 6% vs. 46 +/- 9%; P = 0.017, Cohen's d = 2.3) and CF values (35 +/- 10% vs. 55 +/- 9%; P = 0.025, d = 1.7). Furthermore, trehalose preconditioning reduced perivascular edema, myocardial oxidative stress, and apoptosis. The eNOS dimerization ratio was increased by trehalose (1.2 +/- 0.2 vs. 1.6 +/- 0.2; P = 0.023, d = 2.1), which was associated with the recovery of RPP and CF. These effects of trehalose were abolished by the eNOS inhibitor. Trehalose preconditioning showed protective effects on cardiac and coronary endothelial function after I/R through the eNOS signaling pathway

Cardioprotective effects of chloroquine pretreatment on ischemic and reperfusion injury via activation of ERK1/2 in isolated rat hearts

Purpose Several therapeutic agents have been found to prevent myocardial ischemic and reperfusion (I/R) injury after cardiac surgery; however, no drug is routinely used to afford cardioprotective benefits in clinical settings. Herein, we aimed to determine whether chloroquine (CQ) pretreatment attenuates I/R injury after global ischemia in isolated rat hearts and elucidate mechanisms underlying the effects of CQ. Methods Isolated rat hearts were subjected to 30-min global ischemia, followed by 60-min reperfusion with Krebs-Henseleit buffer (KHB). Immediately before ischemia, 10 mL of pretreatment solutions (KHB, n = 4 or KHB + CQ [100 mu M], n = 4) were injected through the aortic root. Cardiac function was examined based on the rate pressure product (RPP). Myocardial apoptosis was evaluated using TUNEL staining. To assess the reperfusion ischemia salvage kinase pathway, protein expression levels of AKT and extracellular signal-regulated kinase (ERK1/2) were determined using western blotting. To investigate the role of ERK1/2, an ERK1/2 selective inhibitor was used in eight additional rats. Results The recovery rate of the RPP was higher in the KHB + CQ group than in the KHB group 60 min after I/R (KHB, 44 +/- 3% vs. KHB + CQ, 69 +/- 7%; P = 0.019, d = 2.2). CQ pretreatment reduced apoptosis and enhanced the phosphorylation of ERK1/2; however, AKT phosphorylation was unaltered. In addition, the ERK1/2 inhibitor abolished CQ-mediated cardioprotective effects. Conclusions CQ pretreatment showed protective effects on cardiac function after I/R by activating ERK1/2

A simple closure method for a mechanical aortic valve in left ventricular assist device implantation

Because a mechanical aortic valve is a contraindication for the implantation of left ventricular assist device, complicated additional procedures such as a replacement with a bioprosthesis and a closure of the left ventricular outflow tract are required to implant the device. Among such procedures, a sandwich plug technique using vascular clips is one of the simple and feasible procedures. However, this technique requires an off-label use of vascular clips within the aorta that could be associated with a risk of dislodgement and embolization. Thus, we developed a modified sandwich technique without using vascular clips, where the valve leaflets were fixed in the closed position using felt patches and sutures instead of vascular clips. This modified technique is a simple and secure method to close the mechanical aortic valve with the minimum use of artificial materials

Trehalose preconditioning for transient global myocardial ischemia in rats

Autophagy is an intracellular pathway that degrades unnecessary proteins and organelles and provides energy substrates during cellular ischemic conditions. Although pharmacological myocardial pre -conditioning with an autophagy inducer has been reported to protect cells against ischemic reperfusion (I/R), the effects of preconditioning using naturally occurring substances are still unknown. We aimed to examine whether autophagic preconditioning with trehalose improves cardiac function after myocardial stunning by global ischemia in rats. Rat hearts were perfused by oxygenized Krebs Henseleit (KH) so-lution in Langendorff system. Ten rats were randomized into the following two groups according to the perfusates during the preconditioning: control (KH solution only, n = 5) and trehalose (KH & thorn; 2% trehalose, n = 5). After the 35-min preconditioning period and subsequent 20 min of global ischemia, the hearts were reperfused for 60 min. Cardiac function was assessed during the reperfusion. To evaluate autophagy, myocardial protein expression of microtubule-associated protein light chain 3 (LC3) II was evaluated by western blotting. During I/R, a systolic functional parameter, maximum dP/dt was signifi-cantly higher; meanwhile, coronary flow tended to be higher in the trehalose group than in the control group. Myocardial LC3-II expression after preconditioning was higher in the trehalose group than in the control group and decreased to the control level after I/R. In conclusion, in a rat model of global myocardial ischemia, trehalose preconditioning improved cardiac function during I/R. Further studies would be needed to identify the mechanism and effects of trehalose preconditioning. (c) 2021 Elsevier Inc. All rights reserved