Biomedical subjects
Publications and source records attributed to M S Alam.
With the AIDS epidemic continuing to spread throughout the world, development of a safe, practical, and effective HIV vaccine is a national priority. HIV vaccine research efforts are currently targeted towards design of HIV immunogens that induce both cellular and humoral immunity. This brief review summarizes ongoing work at the Duke University School of Medicine on HIV vaccine development.
In this study, we show that the toxicity of ferric nitrilotriacetate (Fe-NTA) can be correlated with the tissue accumulation of 4-hydroxy-2-nonenal (HNE)-modified protein adducts. It is observed that the toxic manifestations of Fe-NTA gradually increase with the increasing age of animals. A dose of Fe-NTA which produces almost 100% mortality in aged rats causes 70% mortality in adults, 30% in pups, 20% in litters, and less than 10% in neonates. The age-dependent increase in its toxicity is also evident from the data of renal microsomal lipid peroxidation and hydrogen peroxide generation. No significant difference in the generation of H2O2 and induction of renal microsomal lipid peroxidation between saline- and Fe-NTA-treated neonates, litters, and pups could be observed. However, in adult rats, a significant increase in both of the parameters was observed which was even greater in aged rats. On the contrary, renal glutathione levels in these animals show an inverse relationship with the oxidant generation. In neonates, litters, and pups the maximum decrease of glutathione was up to 22%, whereas in adult and aged rats, the depletion was more than 60% of their respective saline-treated controls. Parallel to this data, blood urea nitrogen and creatinine, the indicators of renal damage, show a significant increase in Fe-NTA-treated adult and aged rats only, whereas no significant alterations were observed in other groups. Similarly, the magnitude of ODC induction and [3H]thymidine incorporation was much higher in aged and adult rats in comparison to other groups of animals after Fe-NTA treatment. Additionally, the immunohistochemical localization studies show a significant increase in HNE-modified protein adducts in kidney of adult and aged rats, whereas no significant staining was observed in other groups. A similar increase in the level of protein carbonyls has also been observed with the increasing age of rats. These data suggest that the toxicity of Fe-NTA increases with the increasing age of rats and correlates with the accumulation of HNE-modified protein adducts. It may also be speculated that Fe-NTA-mediated renal toxicity leading to carcinogenesis may be related to the tissue accumulation of HNE-modified protein adducts. However, further studies are needed to establish a definite role of HNE-modified proteins in Fe-NTA-mediated carcinogenesis.
OBJECTIVE: Recent increases in the sensitivity of methods used to measure TSH, especially third generation assays, have enabled separation of partial from complete pituitary suppression in patients with thyrotoxicosis. We have investigated the use of a sensitive chemiluminescent enzymeimmunoassay in the differential diagnosis of thyrotoxicosis. DESIGN AND PATIENTS: Serum TSH concentrations were determined by chemiluminescent enzymeimmunoassay in patients with various types of overt and subclinical thyrotoxicosis. RESULTS: The assay was highly sensitive with an analytical sensitivity of 0.0016 mU/l. Among 45 hyperthyroid patients with untreated Graves' disease, 37 (82.2%) showed undetectably low levels (< 0.002 mU/l). Serum TSH in the remaining 8 patients (17.8%) was 0.003-0.005 mU/l. In contrast, TSH was undetectably low in only 5 (20.0%) of 20 patients with painless thyroiditis and in 2 (12.5%) of 16 patients with subacute thyroiditis. Eleven (55.0%) with painless thyroiditis and 12 (75.0%) with subacute thyroiditis had TSH values greater than 0.005 mU/l (0.006-0.032 and 0.006-0.228 mU/l, respectively; normal range 0.5-5.0 mU/l). Serum TSH levels were subnormal in 10 of 12 patients with euthyroid ophthalmic Graves' disease, including 4 with undetectably low levels. Among 11 patients with an autonomously functioning thyroid nodule 6, including 1 with normal free T4 and free T3 and 2 with normal free T3, showed TSH values less than 0.002 mU/l. No significant correlation was observed between serum free T4 or free T3 and TSH concentrations in thyrotoxic patients. Together with the incomplete suppression of TSH observed in those with destructive thyroiditis, this suggests that the grade of TSH suppression was influenced by the duration of illness at the time of blood sampling. CONCLUSION: The third-generation TSH assay is useful for the differential diagnosis of various types of thyrotoxicosis, especially between Graves' disease and destructive thyroiditis.
Recently, we have reported that a new synthetic compound, 1,2bis(nicotinamido)-propane (nicaraven), improved cardiac function following preservation and reperfusion. In this study, we investigated the efficacy of nicaraven as a radical scavenger by using an in vitro model of oxidative stress, to clarify mechanisms of the protective effect of this new compound on reperfusion injury in rat heart. Ring segments of epicardial right coronary arteries (RCA) of pig were suspended in organ chambers and exposed to hydroxyl radicals (.OH), generated (by two different systems) by 0.28 mM FeSO4/0.28 mM H2O2 and DHF/Fe3+-ADP (2.4 mM, 43 nM, and 1.56 uM, respectively) to the bathing solution for 60 min. Prior exposure of the coronary arteries to .OH significantly produced right-ward shift of the dose-response curves of the bradykinin-induced endothelium-dependent relaxations (an increase in the ED50 value for bradykinin by 4.37 and 1.98 times than control in two different .OH generating systems, respectively), but did not affect the maximum relaxation responses. The presence of nicaraven (10(-4) and 10(-5) M) in the .OH generating system, shifted the dose-response curves to bradykinin to the control level, suggesting a significant hydroxyl radical scavenging effect of the drug. These results indicate that nicaraven, a new hydroxyl radical scavenger, exhibits a protective effect on hydroxyl radical-induced endothelial dysfunctions of pig coronary artery.
Technetium-99m methoxyisobutyl isonitrile (99mTc-MIBI) scintigraphy has recently been used in clinical application for detecting thyroid cancer metastases, its role being considered supplementary to serum thyroglobulin (Tg) measurements and radioactive iodine (131I) whole-body scans (WBS). The present retrospective study was designed to elucidate the role of 99mTc-MIBI scans in localizing metastatic lesions by assessing sensitivity and specificity of the scan results obtained in a group of 68 thyroidectomized thyroid cancer patients. Presence or absence of thyroid cancer was judged with other diagnostic modes including serum Tg measurements, 131I WBS, bone scans, chest x-rays, computed tomography (CT), ultrasonography, histopathology, and evolution of disease during follow-up. All scans were read on lesion basis for detecting neck, lung, and bone metastases and also on region basis, namely head-neck, chest, and abdomen-pelvis-extremities (ab-p-ex) areas. The sensitivity of detection was 94.4% (17/18) for neck, 78.4% (40/51) for lung, and 92.8% (64/69) for skeletal lesions. Positive predictive value (PPV) and negative predictive value (NPV) were 96.3% (26/27) and 97.7% (43/44) for head-neck; 94.7% (71/75) and 50.0% (12/24) for chest; 100.0% (25/25) and 93.1% (54/58) for ab-p-ex regions, respectively. For all scan sites taken together, PPV and NPV were 96.1% (122/127) and 86.5% (109/126), respectively. In conclusion, the present study reveals that 99mTc-MIBI can be proposed as a first-line diagnostic agent for the follow-up protocol of thyroid cancer patients, although the ability to detect small lung metastases is somewhat limited.
BACKGROUND: The purpose of this study was to compare the efficacy of histidine-tryptophan-ketoglutarate (HTK) solution after prolonged cold storage with that of the conventional glucose-insulin-potassium (GIK) and University of Wisconsin (UW) solutions in experimental heart preservation. GIK solution was chosen as a control to mimic current clinical regimens. Variables of cardiac function, myocardial tissue water, and adenine nucleotide pool metabolites were used to assess prolonged myocardial preservation in the isolated rat heart model. METHODS: Hearts isolated from male Wistar rats were mounted on a Langendorff apparatus to estimate baseline cardiac function. The hearts were divided into three groups (n=6 per group) according to each preservation solution used: group 1, GIK solution; group 2, UW solution; and group 3, HTK solution. The hearts were then arrested and stored in each solution for 6, 8, and 12 hr at 4 degrees C. After storage, the hearts were reperfused and recovery of cardiac function and myocardial tissue water content were evaluated. Myocardial adenylate contents just after storage in each group (n=5 hearts/group) were also measured. RESULTS: The hearts stored in HTK solution showed maintenance of cardiac function at up to 8 hr of almost 80% of prepreservative baseline function; however, recovery of cardiac function of the hearts stored in UW solution revealed an initial loss of function at 6 hr of almost 60% and a decline to 50% at 8 hr. Furthermore, recovery of cardiac function of the hearts stored in GIK solution revealed a progressive loss of function at 6 hr of storage of almost 50% and a decline to 30% at 8 hr of storage. The myocardial ATP/ADP ratios after 6 hr of storage in HTK and UW solutions were significantly higher than the ratio found in GIK solution. Although the myocardial ATP/ADP ratio after 8 hr of storage in HTK solution was maintained above 50%, the ratios in GIK and UW solutions declined to 15%. In addition, the myocardial energy charge values of the hearts stored in HTK solution were sufficiently maintained until 8 hr of storage, whereas the values in UW and GIK solutions declined to below 50% at 6 hr of storage and 20% at 8 hr of storage. Although there were no significant differences in tissue water contents after 6 and 8 hr of storage among the three groups, the water contents of the hearts after 12 hr of storage in HTK and UW solutions were significantly lower than that of the hearts stored in GIK solution. CONCLUSIONS: Our results suggest that HTK solution is much more effective than UW and GIK solutions for isolated rat heart preservation; however, successful cold storage of the heart is highly energy-dependent, and a dramatic breakdown of myocardial energy level, which causes a crucial decline in cardiac function, occurs between 8 and 12 hr of storage.
BACKGROUND: The effects of a soybean oil diet and a high-cholesterol oil (HC) diet, and an HC diet with eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) supplementation, on basal and postpreservative cardiac function of the hearts and on postpreservative renal function of the kidneys from older rats were examined. METHODS: Groups 1 through 4 of 100-week-old rats were fed either soybean oil, HC, HC with EPA, or HC with DHA, respectively, for 12 weeks. Blood was collected for analysis of plasma fatty acids, and the heart and left kidney were removed from the rat. In experiment 1, the heart was perfused on a Langendorff apparatus. After evaluation of the cardiac function of each rat, the heart was stored in histidine-tryptophan-ketoglutarate solution for 8 hr at 4 degrees C. The heart was reperfused and the recovery of cardiac function was evaluated. The coronary perfusate during reperfusion was collected to measure 6-keto prostaglandin F1alpha and thromboxane B2. Coronary flow (CF) perfused with Krebs-Henseleit bicarbonate (KHB) solution containing 5-hydroxytryptamine (5-HT) and nitroglycerin were evaluated in the Langendorff mode with atrial pacing (330 beats/min). In experiment 2, the excised left kidney was immediately flushed and preserved with University of Wisconsin solution for 8 hr at 4 degrees C. The kidney was then reperfused with KHB solution and renal function was evaluated. RESULTS: The plasma and cardiac EPA levels in group 3 were significantly higher than the levels found in the other groups. The plasma and cardiac ratios of EPA to arachidonic acid were significantly higher in groups 3 and 4 than in groups 1 and 2. There were no significant differences in basal cardiac function among any of the diet-fed rats. The percentage values of the recovery of aortic flow, cardiac output (CO), and left ventricular max dp/dt in group 3 and CO in group 4 were significantly higher than in group 2. In addition, the recovery of CF in group 3 tended to be higher than in group 2 (P=0.07). The percentage values of the recovery of aortic flow, CF, CO, and left ventricular max dp/dt in group 1 were significantly lower than in the other dietary groups. CF reperfused with KHB solution containing 5-HT was significantly higher in group 3 than in groups 1 and 2. CF reperfused with KHB solution containing 5-HT was significantly higher in group 4 than in group 1. CF reperfused with KHB solution containing nitroglycerin in group 3 tended to be higher than in groups 1 and 2 (P=0.07). The thromboxame B2 concentrations in the coronary perfusate during reperfusion in groups 3 and 4 were significantly lower than in groups 1 and 2. Fractional sodium reabsorption in group 3 was significantly higher than in group 2. Inulin clearance in groups 3 and 4 was significantly higher than in group 1. The postpreservative urinary flow in group 3 was significantly higher than in groups 1 and 2. The urinary flow was significantly higher in group 4 than in group 1. CONCLUSIONS: These results suggest that EPA administration may attenuate preservation and reperfusion injury and improve the recovery of cardiac and renal functions in hyperlipidemic and older rats. DHA administration may also show beneficial effects on kidney preservation in hyperlipidemic rats.
To develop a new method of radioimmunodetection for thyroid cancer, we tested the binding ability of a human antithyroglobulin monoclonal antibody, VB5, to primary culture of human thyroid cancer cells. VB5 was able to immunostain cytoplasmic thyroglobulin (Tg) in the acetone-fixed cancer cells when used in a labeled streptavidin-biotin method but not in a conventional indirect immunoperoxidase technique. The antibody was readily labeled with I-125 in the standard chloramin-T method, and showed specific binding to the antigen on cultured malignant thyrocytes displaceable with non-labeled VB5 or with excess Tg antigen. Although these initial results in vitro are encouraging, the observed low specific binding (about 1% at room temperature) to intact cells with a single monoclonal antibody seems insufficient to conduct any in vivo immunolocalization experiments in animals. To obtain more binding, we would need a cocktail of several monoclonal antibodies to different epitopes, and also fragmentation of antibody molecules to penetrate into cytoplasm.
Pure geometric isomers of conjugated linoleic acid were prepared from castor oil as the primary starting material. Methyl octadeca-9Z,11E-dienoate (2) and methyl octadeca-9Z,11Z-dienoate (4) were obtained by zinc reduction of methyl santalbate (1, methyl octadec-11E-en-9-ynoate) and methyl octadec-11Z-en-9-ynoate (3), respectively, as the key intermediates. Methyl octadeca-9E,11E-dienoate (8) and methyl octadeca-9E,11Z-dienoate (9) were prepared by demesylation of the mesyloxy derivative of methyl ricinelaidate (6, methyl 12-hydroxy-octadec-9E-enoate). A study of the nuclear magnetic resonance spectral properties was carried-out, and the shifts of the olefinic carbon atoms of 18:2(9Z,11E) (2) and 18:2(9E,11Z) (9) were readily identified by a combination of incredible natural abundance double quantum transfer experiment, heteronuclear multiple bond correlation, and 1H-13C correlation spectroscopy correlation techniques. Doubts remain in the absolute identification of the individual olefinic carbon atoms of the 18:2(9Z,11Z) (4) and 18:2(9E,11E) (8), except the fact that the shifts of the "inner" (C-10 and C-11) and "outer" (C-9 and C-12) positioned olefinic carbon atoms of the conjugated diene system are distinguishable.
Reaction of methyl undec-10-ynoate (1) with selenium dioxide/tert-butyl hydroperoxide (TBHP) in aqueous dioxane gave methyl 9-oxo-undec-10-ynoate (2, 9%) and 9-hydroxy-undec-10-ynoate (3, 60%), while methyl octadec-9-ynoate (4) yielded mixtures of positional isomers of mono-keto (viz. methyl 8-oxo- and 11-oxo-octadec-9-ynoate, 5, 5%), hydroxy-keto (viz. methyl 8-hydroxy-11-oxo- and 11-hydroxy-8-oxo-octadec-9-ynoate, 6, 10%), and dihydroxy (viz. methyl 8,11-dihydroxy-octadec-9-ynoate, 7, 24%) derivatives. Similar treatment of a conjugated diacetylenic fatty ester (methyl octadeca-6,8-diynoate, 8) furnished a mixture of methyl 5-oxo- and 10-oxo-octadeca-6,8-diynoate (9, 12%) and a complex mixture of very polar products. Reaction of methyl octadec-11E-en-9-ynoate (methyl santalbate) (10) with selenium dioxide/TBHP in aqueous dioxane gave exclusively a mixture of regiospecific products, viz. methyl 8-oxo-octadec-11(E)Z-en-9-ynoate (11, 6%) and methyl 8-hydroxy-octadec-11E-en-9-ynoate (12, 70%). The structures of the various products were determined by a combination of spectroscopic and mass spectral analyses.
OBJECTIVE: We investigated the efficacy of nicaraven in reducing myocardial as well as coronary endothelial preservation-reperfusion (P/R) injury. METHODS: In experiment I, isolated rat hearts were mounted on a Langendorff (L) apparatus to estimate the baseline cardiac function. Group 1, 8- and 12-h storage in histidine-tryptophan-ketoglutarate (HTK) solution; group 2, 8- and 12-h storage in HTK solution with superoxide dismutase (2.5 x 10(5) U/I) and catalase (2 x 10(5) U/I); group 3, 8- and 12-h storage in HTK solution with nicaraven (10(-3) M). Following storage for 8 and 12 h at 4 degrees C, they were reperfused and post-preservative cardiac function was evaluated. The hearts were then switched back to L-mode and paced at 330 beats/min. Coronary flow (CF) following perfusion with KHB solution containing 5-hydroxytryptamine (5-HT) and nitroglycerin was also measured. In experiment 2, segments of pig coronary artery were suspended in organ chambers and exposed to hydroxyl radicals in the presence or absence of nicaraven. The sensitivity of relaxation response to bradykinin of the prior-exposed rings was measured. RESULTS: The recovery of CF and LV dp/dt following 8 h of storage in group 3 was higher than that in group 1, although there were no significant differences in the other parameters of cardiac functional recovery among the groups. The absolute values of cardiac function following 12 h of storage in groups 1-3 were as follows: 6.6 +/- 2.4, 9.1 +/- 0.8, 15.6 +/- 3.1 ml/min of cardiac output (CO); 1.9 +/- 0.9, 2.3 +/- 0.4, 6.0 +/- 2.3 ml/min of aortic flow (AF); 4.5 +/- 1.3, 6.0 +/- 0.5, 9.5 +/- 0.8 ml/min of CF, respectively. The recovery of CO, AF, CF, SP, MP, and left ventricular (LV) dp/dt were significantly improved in group 3, compared with those in group 1. CF and CO in group 3 were higher than those in group 2.5-HT caused vasoconstriction in all groups, but the vasoconstriction in group 3 was less than in group 1. Prior exposure to FeSO4/H2O2 produced significant endothelial damage as reflected by the right-ward shift of the dose-response curve of bradykinin-induced endothelium-dependent relaxation. In the presence on nicaraven, the dose-response curve recovered to the control level. CONCLUSIONS: Nicaraven may improve coronary endothelial and myocardial function following P/R by its hydroxyl radical scavenging activity.
Detectability of bone metastases from differentiated thyroid carcinoma by technetium-99m hydroxymethylene diphosphonate ([99m]Tc-HMDP) bone scan is considered to be poor. Thallium-201 (201Tl) is also widely used for detecting metastatic lesions. Our present study was aimed at the evaluation of the combined use of (99m)Tc-HMDP and 201Tl imaging in successful detection of bone metastases from differentiated thyroid carcinoma. Twenty-seven thyroidectomized thyroid cancer patients (19 females, 8 males; 12 papillary type, 15 follicular type) with 77 bone lesions were included in this retrospective study. All of these patients received ablative doses of radioiodine. Thyroidal origin of the lesions was proved by positive iodine-131 (131I) uptake. In 131I-negative lesions, histological proof or absence of tumor markers other than thyroglobulin was considered when computed tomography (CT) and/or magnetic resonance imaging (MRI) suggested metastatic nature of the lesions. Of the 77 lesions, 58 (75.3%) were positive and 19 were negative in the (99m)Tc-HMDP bone scintigraphy, whereas 53 lesions (68.9%) could be detected by 201Tl scintigraphy. However, within the 19 (99m)Tc-HMDP-negative lesions, 14 showed abnormal accumulation of 201Tl, and within the 24 201Tl negative lesions, 19 were positive in (99m)Tc-HMDP scan. This resulted in a combined sensitivity of 93.5%. Our present study concludes that combined (99m)Tc-HMDP and 201Tl imaging is a sensitive and effective method for detecting bone metastases from thyroid carcinoma.
OBJECTIVE: To develop a novel bioassay for the assessment of thyroid cell growth stimulating activity using cytochalasin B (CB) and to test immunoglobulins (IgGs) from patients with autoimmune thyroid diseases. DESIGN: The assay is based on the principle that growing cells during incubation with CB show an increased number of nuclei in a cell (N/C index), since CB, at appropriate concentrations, is known to inhibit cytoplasmic cleavage without affecting nuclear mitosis. The N/C index represents potential DNA production while cells are incubated with CB. METHODS: FRTL-5 thyroid cells were incubated with various thyroid stimulators in TSH-free medium containing 2 mg/J CB for 3 days. After the incubation, the cells were harvested in trypsin/EDTA to obtain single cell suspension, fixed, dropped onto a glass slide, stained and observed under a microscope to determine the N/C index. RESULTS: Bovine TSH at 10(-3)-1.0 U/I, forskolin at 1x10(-7)-10(-5) mol/l, cholera toxin at 10x10(-5)-10(-3) mg/l, or (Bu)2cAMP at 1 x 10(-5)-10(-3) mol/l increased the N/C index up to approximately 2.0 in a dose-dependent manner. IgGs not only from 27 patients with untreated goitrous Graves' disease but also from 14 patients with goitrous Hashimoto's thyroiditis elicited an increase in the N/C index, which exceeded the mean + 2 S.D. of the values for 17 normal subjects (mean +/- S.D., 1.063 +/- 0.014). Four patients with primary myxedema displayed a normal N/C index. In Graves' disease, the N/C index did not correlate significantly with thyroid stimulating antibodies (TSAb) activities but did correlate significantly with estimated goiter size (P < 0.05). IgGs containing blocking-type TSH-receptor antibodies inhibited the TSH- or Graves IgG-stimulated increase in N/C index almost completely, but did not influence the stimulatory effect of IgG from two patients with Hashimoto's thyroiditis. CONCLUSIONS: We have developed a sensitive and simple assay for thyroid growth stimulating activity by using CB, and found that all tested patients with goitrous Graves' disease and goitrous Hashimoto's thyroiditis have thyroid growth stimulating immunoglobulins whose activity does not correlate with TSAb.
UNLABELLED: Technetium-99m-methoxyisobutyl isonitrile (99mTc-MIBI) was evaluated for its ability to detect metastases from thyroid carcinoma. METHODS: Twenty-seven thyroidectomized patients with metastatic differentiated thyroid carcinoma, of whom 20, 9 and 12 had lung, lymph node and bone metastases, respectively, were examined with 99mTc-MIBI. The scan results were compared with those of 201TI and 131I whole-body scans. RESULTS: Increased accumulation of 99mTc-MIBI was observed in lung metastases of 15 patients (75.0%), 12 lymph node metastases (100.0%) and 29 of 31 bone metastases (93.5%). Increased accumulations of 201TI and 131I scans were seen in, respectively, 16 (80.0%) and 17 (85.0%) of the 20 patients with lung metastases, 12 (100.0%) and 5 (41.7%) of the 12 lymph node metastases and 28 (90.3%) and 27 (85.1%) bone metastases. Because of its better image quality, 99mTc-MIBI detected more lesions in the lung (n = 38) than 201TI did (n = 17). CONCLUSION: Technetium-99m-MIBI is clinically useful for detecting metastases from differentiated thyroid carcinoma and deserves clinical application in the postoperative follow-up of such patients.
The "standard" values of pulmonary function tests obtained from Western populations do not agree with that of people of Bangladesh. The present survey was conducted with a view to determining the values of three lung functions parameters, viz., vital capacity, forced vital capacity, and forced expiratory volume in healthy Bangladeshi male population. 250 non-smoker male subjects of age 20-40 years were selected for the study. Of these 150 were university students and employees and 100 were low income group slum dwellers. All three values correlated positively with height and weight and negatively with age. There was a definite effect of aging on the lung function showing a declining trend from age 35 years onwards. The slum dwellers showed lower values (-12.5% in young adults) of lung functions than the university students and employees. The difference decreased with age. Prediction equations were set up using height, weight, and age as parameters. In general the lung function values were found to be 80% of the European Standards.
We investigated the efficacy in reducing myocardial preservation and reperfusion (P/R) injury of direct hydroxyl radical scavenging by nicaraven as compared with scavenging of both superoxide radicals and hydrogen peroxides by superoxide dismutase (SOD) and catalase (CAT), respectively. Isolated rat hearts were mounted on a Langendorff (L) apparatus to estimate the baseline aortic flow (AF), coronary flow (CF), cardiac output (CO), systolic pressure (SP), aortic mean pressure (MP), rate pressure product, and LV dp/dt. They were divided into 3 groups: group 1, 12 hr storage in HTK solution; group 2, 12 hr storage in HTK solution containing 2.5x10(5) U/L SOD and 2x10(5) U/L mg/L CAT; and Group 3, 12 hr storage in HTK solution containing 10(-3) M nicaraven. SOD, CAT, and nicaraven were administered intraperitoneally before harvesting. Hearts were stored in each preservation solution at 4, and then reperfused. Postpreservative function and concentrations of leaked enzymes were measured. The hearts were switched back to the L-mode and paced at 330 beats/min. CF following perfusion with Krebs-Henseleit bicarbonate buffer (KHB) solution containing 10(-6) M 5-hydroxytryptamine (5-HT) or 10(-5) M nitroglycerin (NTG) then evaluated. The myocardial water content also was measured. The recovery of CF, CO, SP, MP, and LV dp/dt was significantly greater in group 3 than in group 1. The recovery of CF was superior to that in group 2 (P<0.05). There were no significant differences in the recovery of cardiac function between groups 1 and 2. 5-HT caused a decrease in CF in each group, however, CF in group 3 was higher than that in group 1 (P<0.05). NTG caused no significant differences among the groups. There were no significant differences in leaked enzymes and myocardial water content among the three groups. These results suggest that nicaraven protects against myocardial P/R injury through its hydroxyl radical scavenging activity, and that therapy with oxygen-free radical scavengers should be directed toward inactivation of hydroxyl radicals rather than superoxide radicals and/or hydrogen peroxides.
We examined the effects of supplementation with eiosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), major components of omega-3 polyunsaturated (correction of polyunsatulated) fatty acids (PUFAs), on basal cardiac function and recovery of cardiac function of "donor hearts" from adults (30 week) rats following cold preservation and reperfusion (P/R). In groups 1, 2, 3, and 4, respectively, 30-week-old rats were fed a soybean oil diet, a high-cholesterol oil (HC) diet, an HC diet with EPA, or an HC diet with DHA for 5 weeks. After collecting blood to analyze plasma levels of fatty acids among each group, the heart was excised and perfused on a Langendorff apparatus. Following evaluation of each rat's cardiac function, each heart was stored in HTK solution for 8 hr at 4 degrees C. The heart was then reperfused and the coronary perfusate was collected to evaluate enzyme that had leaked. After cardiac functional recovery was estimated, myocardial fatty acids were measured. EPA supplementation significantly increases the plasma and cardiac levels of EPA as well as the ratio of EPA to arachidonic acid (AA). EPA supplementation also led to improved recovery of cardiac function following P/P, compared with that of rats who received soybean oil, high-cholesterol oil, and DHA. DHA supplementation significantly increased the plasma and cardiac levels of DHA as well as the ratio of DHA to AA--however, the cardiac functional recovery was almost identical to that of the rats who received high-cholesterol oil and was higher only than that of the rats who received soybean oil. There were no significant differences in enzyme that had leaked and myocardial water content among each group. These results suggest that alterations in the myocardial phospholipid composition by EPA supplementation may be profoundly responsible for attenuating myocardial I/R injuries. In contrast, DHA supplementation may not exert a cardioprotective effect following cold P/R. DHA supplementation alone may not increase the myocardial level of EPA enough to cause a protective effect against P/R injury. EPA supplementation to hyperlipidemic patients may be clinically warranted for increasing the potential number of donors.
A comparative study on the level of expression of lacZ reporter constructs driven by equivalent carp and rat beta-actin regulatory sequences was carried out in embryos of tilapia and rainbow trout. DNA was microinjected into fertilised tilapia and rainbow trout eggs and the embryos/fry were assayed at various developmental stages for beta-galactosidase expression. We provide evidence to demonstrate that the carp beta-actin promoter/ lacZ reporter gene is expressed at higher levels than the equivalent rat beta-actin construct in both species.