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M M Sayeed

Publications and source records attributed to M M Sayeed.

At least 55 records · Page 3Linked to original sources
PubMed1994

T-lymphocyte Ca2+ signalling and proliferative responses during sepsis.

Alterations in T-lymphocyte kinetics and/or activation have been implicated in burn and traumatic injury. This study evaluated concanavalin A (Con A) regulation of both intracellular Ca2+ (Cai2+) and proliferation in T-lymphocytes harvested from spleens of septic rats. Rats were implanted with fecal pellets containing Escherichia coli (150 colony forming units (CFU)) and Bacteroides fragilis (10(4) CFU). T-cell[Ca2+]i was measured before and after treatment of cells with ConA, using Fura-2 and microfluorophotometry. Splenic lymphocytes were cultured for 72 h with Con A to assess their proliferative response. As compared to sterile-implanted rats, the septic rat T-lymphocytes Cai2+ response to Con A significantly decreased on days 1 and 2 after implantation. A significant decrease in T-cell proliferative response to Con A, compared to sterile controls, was found in septic rats on day 2 but not on day 1. These results suggest that Con A-mediated proliferation in T-cells occurs secondarily to a decrease in cellular Ca2+ signalling. The depression in the T-cell proliferative response during sepsis could contribute to a decrease in host's resistance against sepsis.

Animals↗
PubMed1994

T-lymphocyte Ca2+ signalling and proliferative responses during sepsis.

Alterations in T-lymphocyte kinetics and/or activation have been implicated in burn and traumatic injury. This study evaluated concanavalin A (Con A) regulation of both intracellular Ca2+ (Ca2+i) and proliferation in T-lymphocytes harvested from spleens of septic rats. Rats were implanted with fecal pellets containing Escherichia coli (150 colony forming units (CFU)) and Bacteroides fragilis (10(4) CFU). T-cell [Ca2+]i was measured before and after treatment of cells with Con A, using Fura-2 and microfluorophotometry. Splenic lymphocytes were cultured for 72 h with Con A to assess their proliferative response. As compared to sterile-implanted rats, the septic rat T-lymphocytes Ca2+i response to ConA significantly decreased on days 1 and 2 after implantation. A significant decrease in T-cell proliferative response to Con A, compared to sterile controls, was found in septic rats on day 2 but not on day 1. These results suggest that Con A-mediated proliferation in T-cells occurs secondarily to a decrease in cellular Ca2+ signalling. The depression in the T-cell proliferative response during sepsis could contribute to a decrease in host's resistance against sepsis.

Animals↗
PubMed1994

In vitro antitumor activity of cromakalim in human brain tumor cells.

The effect of cromakalim, a K+ channel opener, on the growth of human brain tumor cells was investigated. Cromakalim inhibited the growth of SK-N-MC human neuroblastoma and U-373 MG human astrocytoma cell lines in a dose-dependent manner. This effect of cromakalim was significantly blocked by the co-treatment with sulfonylureas (glibenclamide or tolbutamide) which are known as specific blockers for ATP-sensitive K+ channels. In addition, cromakalim significantly inhibited agonist-induced intracellular Ca2+ mobilization in the astrocytoma cells. This inhibition induced by cromakalim was also reversed by pretreatment with glibenclamide. These results suggest that the antitumor activity of cromakalim may be due to the activation of ATP-sensitive K+ channels leading to the inhibition of the intracellular Ca2+ signalling mechanism.

Antineoplastic Agents↗
PubMed1993

Inhibition of human brain tumor cell growth by a receptor-operated Ca2+ channel blocker.

SK&F 96365, a reported receptor-operated Ca2+ channel blocker, inhibited the growth of U-373 MG human astrocytoma and SK-N-MC human neuroblastoma cell lines in a dose-dependent manner. Carbachol and serum which act as growth factors for these cells induced a rapid, transient increase of intracellular Ca2+ concentration without a sustained increase. SK&F 96365 also exerted a significant inhibition of carbachol or serum-induced intracellular Ca2+ mobilization. These results suggest that SK&F 96365 is a potent inhibitor of brain tumor cell growth and that its effect may be mediated by the inhibition of agonist-induced intracellular Ca2+ mobilization.

Astrocytoma↗
PubMed1993

Altered membrane fluidity in rat hepatocytes during endotoxic shock.

Steady-state fluorescence anisotropy measurements of the fluorescent hydrocarbon probe 1,6-diphenyl-1,3,4-hexatriene (DPH) were carried out in isolated hepatocytes of saline control and Salmonella enteritidis endotoxin (20 mg/kg) injected rats. Statistically significant differences were observed in the fluorescent anisotropy (rs) and membrane microviscosity (eta) values of control (rs = 0.107 +/- 0.004 (SEM), eta = 0.98 +/- 0.08, n +/- 6) versus endotoxin injected rat hepatocytes (rs = 0.134 +/- 0.005, eta = 1.43 +/- 0.08, n = 6, p < 0.001) at 37 degrees C. Fluidity was similarly lower in the isolated plasma membrane preparations from endotoxin-injected rat livers relative to control livers. When endotoxin-injected rats were treated with the calcium channel-blocker diltiazem, the anisotropy and microviscosity values were comparable to those obtained from control rats (rs = 0.152 +/- 0.003, eta = 1.00 +/- 0.003, n = 6). These measurements were made in animals five hours after endotoxin had been injected, and thus represent the in vivo effects of bacterial endotoxins. Temperature scan studies of DPH from 5-40 degrees C revealed that the membrane fluidity of endotoxin-injected rat hepatocytes was significantly lower than control hepatocytes at all temperatures investigated. The data suggest that endotoxin alters the membrane fluidity of hepatocytes, and that calcium-channel blockers can prevent the alteration. Our previous studies have shown that calcium channel blocker prevented endotoxin induced alterations in hepatic cellular regulation of Ca2+. Thus, cellular calcium homeostasis may be important in the maintenance of membrane fluidity and other membrane-associated transport functions.

Animals↗
PubMed1993

Inhibition of cell growth by K+ channel modulators is due to interference with agonist-induced Ca2+ release.

The effects of K+ channel modulators, tetraethylammonium, 4-aminopyridine and diazoxide, and high extracellular K+ on cell growth and agonist-induced intracellular Ca2+ mobilization were investigated. Two human brain tumour cell lines, U-373 MG astrocytoma and SK-N-MC neuroblastoma, were used as model cellular systems. K+ channel modulators and increased extracellular K+ concentration inhibited tumour cell growth in a dose-related fashion in both cell lines. In addition, agonist (carbachol or serum)-induced intracellular Ca2+ mobilization was also blocked by the pretreatment of growth-inhibitory concentrations of K+ channel modulators and high extracellular K+. Thus, these results suggest that K+ channel modulators are effective inhibitors of brain tumour cell growth and that their growth regulation may be due to the interference with the intracellular Ca2+ signalling mechanisms.

4-Aminopyridine↗
PubMed1993

Skeletal muscle Ca2+ flux and catabolic response during sepsis.

Membrane Ca2+ flux and net protein catabolism were studied in the skeletal muscle during experimental sepsis. Sterilized rat fecal pellets with (septic) or without (sterile) gram-negative bacteria, Escherichia coli [10(2) colony-forming units (cfu)] and Bacteroides fragilis (2 x 10(3) cfu), were implanted into the abdomens of male Sprague-Dawley rats (110-120 g). Septic and sterile rats were febrile and hyperlactacidemic on day 1 postimplantation. These responses subsided by day 2 in sterile but not septic rats. Initial Ca2+ flux, estimated from measurements of 45Ca uptake by soleus muscles in vitro, was elevated on day 1 in both sterile and septic rats and on day 2 and 3 in septic rats only. The septic rat soleus muscle showed a significantly increased net protein catabolic response (measured as tyrosine release by soleus muscle, in vitro) over that found in muscles of sterile rats on day 1-3 postimplantation. The increase in Ca2+ flux in septic (day 1-3 postimplantation) and sterile (day 1 only) rats was abolished when the rats were treated with the calcium channel blocker diltiazem. In unoperated control rat soleus muscles the Ca2+ ionophore, ionomycin, concomitantly caused an increase in Ca2+ flux and net protein catabolism. Overall, the present study suggested that altered cellular Ca2+ regulation plays a role in the net protein catabolic response in the skeletal muscle during sepsis.

Animals↗
PubMed1992

Ca(2+)-related hepatocellular alterations during intra-abdominal sepsis.

Intra-abdominal sepsis was induced in rats by implanting into their abdominal cavities fecal-agar pellets impregnated with Escherichia coli and Bacteroides fragilis. Sham-operated rats received sterile pellets. A group of sterile- and septic-implanted rats was treated intraperitoneally with diltiazem (1.2 mg/kg) 8 h after implantations. Septic- and sterile-implanted rat hepatocytes were loaded with 1) the fluorescent dye indo-1 to quantify hepatocyte basal and vasopressin (100 nM)-elevated cytosolic Ca2+ concentration and 2) 45Ca to quantify Ca2+ flux and cellular content of exchangeable Ca2+. Lipid peroxidation was determined by measuring conjugated dienes (CD) and thiobarbiturate-reactive substances (TBA-RS) in liver homogenates. In septic-implanted rats, the basal cytosolic [Ca2+], cellular exchangeable Ca2+, Ca2+ flux, CD, and TBA-RS were significantly higher than in sterile-implanted rats. Although vasopressin caused a significant elevation in cytosolic [Ca2+] in septic rat hepatocytes, the magnitude of this elevation was significantly smaller than that found in the sterile group. Diltiazem treatment of septic rats significantly decreased basal cytosolic [Ca2+], cellular exchangeable Ca2+ content, Ca2+ flux, CD, and TBA-RS. Also, vasopressin-induced increase in hepatocyte cytosolic [Ca2+] in diltiazem-treated septic rats was significantly greater than that observed in untreated septic rats. Both Ca2+ and membrane lipid alterations were attenuated with diltiazem treatment of septic rats. These results suggest that prevention or attenuation of Ca2+ channel-mediated Ca2+ influx restores both Ca2+ homeostasis and membrane lipid alteration.

Abdomen↗
PubMed1992

Effect of endotoxic shock on basal and insulin-mediated Na+/K(+)-pump activity in rat soleus muscle.

This study evaluated basal and insulin-mediated Na+/K+ transport in skeletal muscle during endotoxic shock. Fasted male Holtzman rats (80-100 g) were killed 5 hr after the i.v. injection of saline (control) or 20 mg/kg Salmonella enteritidis endotoxin. 86Rb+ uptake into the cellular compartment of soleus muscle was determined in the presence and absence of 100 mU/ml insulin. The rate coefficient and rate of 22Na+ efflux and muscle intracellular 22Na+ content (in percentage of total muscle 22Na+) were determined in the presence and absence of 1 mM ouabain. The rate of basal cellular 86Rb+ uptake (cpm x 10(3).g wet weight-1.min-1) was not significantly different between control (77.7 +/- 2.9) and endotoxic soleus muscles (76.1 +/- 3.9). Insulin increased the rate of cellular 86Rb+ uptake by 19% in control (92.4 +/- 3.7) and 23% in endotoxic soleus muscles (93.8 +/- 4.1). Cellular 22Na+ content was 40% greater in endotoxic muscle (16.8 +/- 1.4) as compared to control muscles (12.0 +/- 0.9). The rate coefficient for ouabain-sensitive 22Na+ efflux in endotoxic muscles (0.028 min-1) was 1.7-fold greater than that in control muscles (0.017 min-1). The rate of ouabain-sensitive 22Na+ efflux was more than doubled in endotoxic muscles (0.470%/min) compared with control muscles (0.204%/min). Endotoxic shock did not alter insulin's ability to stimulate Na+/K+ transport in muscle. An increase in the electrogenicity of the Na+/K+ pump is consistent with an increased rate coefficient for 22Na+ efflux, with no change in the rate of 86Rb+ uptake in endotoxic soleus muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Effect of alpha-adrenergic blockage on cellular Ca2+ during endotoxic shock.

The effect of alpha-adrenergic receptor antagonists, phentolamine, and prazosin on cytosolic Ca2+ concentration, [Ca2+]i, was studied in hepatocytes during endotoxic shock. Rats were given intravenous injections of endotoxin (20 mg/kg), phentolamine (3 mg/kg) plus endotoxin (20 mg/kg), or prazosin (5 mg/kg) plus endotoxin (20 mg/kg). They were sacrificed 5 hr later, at which time the endotoxin-injected rats showed signs of shock. Isolated hepatocytes were prepared and employed for the measurement of [Ca2+]i under basal and hormone-stimulated (1 and 10 microM epinephrine) conditions by means of Quin 2 fluorescence technique. The apparent basal level of [Ca2+]i in endotoxic rat hepatocytes (mean +/- SE: 482 +/- 31 nM) was significantly higher (P less than 0.05) than in phentolamine plus endotoxin (242 +/- 73) and prazosin plus endotoxin (240 +/- 43) groups. A significant increase in hepatocyte [Ca2+]i occurred with epinephrine in the phentolamine plus endotoxin and prazosin plus endotoxin groups, but not in the group receiving endotoxin alone. Endotoxic rats showed a mortality rate of 75%, whereas phentolamine plus endotoxin and prazosin plus endotoxin groups showed a mortality rate of 38% and 20% respectively. These data suggest that the protective effect of alpha-adrenergic receptor antagonists during endotoxic shock may be mediated, in part, by attenuating the entrance of Ca2+ into endotoxic liver cells.

Animals↗
PubMed1991

Calcium-dependent and calcium-independent protease activities in skeletal muscle during sepsis.

Sepsis was produced in rats by implanting into their abdominal cavities fecal pellets containing Escherichia coli (10(2) colony-forming units [CFU]) and Bacteroides fragilis (10(4) CFU). Control rats were implanted with sterile pellets. A febrile response and hyperlactacidemia marked the onset of the septic injury. Control and septic rats were killed 24 and 48 hr after implantations, and posterior leg muscles were removed. Muscles were homogenized to prepare soluble fractions containing calcium-independent lysosomal (cathepsins B and L) and calcium-dependent cytosolic (calpain) proteases. Cathepsin and calpain activities were then assayed using standard procedures. There were no alterations in cathepsins B or L activities during sepsis. Calpain activity in septic muscle was significantly higher than that in control muscles. In vitro calpain sensitivity to Ca2+ was also higher in septic muscle than in controls. The cysteine protease inhibitor leupeptin caused a quantitatively greater inhibition of calpain activity in septic than in control muscles. These data indicate that whereas sepsis has no effect on Ca(2+)-insensitive lysosomal proteases, it is associated with an elevation of the Ca(2+)-dependent cytosolic protease activity.

Animals↗
PubMed1990

Effect of Ca2(+)-channel agonists and antagonists on skeletal muscle sugar transport.

The purpose of this study was to determine whether changes in cellular Ca2+ caused by Ca2(+)-channel modifiers affect skeletal muscle sugar transport. Rat soleus muscles were isolated, and sugar transport was measured monitoring efflux of 3-O-methylglucose (3-MG). Muscles were "loaded" with 3-O-[methyl-14C]methyl-D-glucose in Krebs-Ringer-bicarbonate (KRB) media and then sequentially washed in radioisotope-free KRB. Cellular Ca2+ was modified by adding a Ca2+ agonist (BAY K 8644) and/or Ca2+ antagonists (nifedipine, nitrendipine, diltiazem) to the "load" and "wash" media. Alterations in cellular Ca2+ were determined from soleus muscle 45Ca2+ uptake measurements. Addition of 0.25 microM BAY K 8644 enhanced insulin-mediated 3-MG transport and 1 microM nifedipine prevented the agonist effect. A high concentration of BAY K 8644 (6 microM) led to increased basal but attenuated insulin-mediated 3-MG transport. Nifedipine (4 microM) and diltiazem (20 microM) blocked the BAY K 8644-induced changes in basal and insulin-mediated 3-MG transport. Nifedipine (4 microM) also attenuated the stimulation of cellular Ca2+ uptake by 6 microM BAY K 8644. Nitrendipine was unable to reverse the changes in basal or insulin-mediated 3-MG transport caused by BAY K 8644. These results indicate that a low concentration of BAY K 8644 enhanced insulin-stimulated skeletal muscle sugar transport, whereas a high concentration of the agonist led to a diminished ability of insulin to stimulate sugar transport. The effects of the Ca2+ agonist were presumably mediated through modulation of cellular Ca2+ as these effects were blunted by the Ca2+ antagonists.

3-O-Methylglucose↗
PubMed1989

Skeletal muscle calcium uptake in bacteremic rats.

To determine whether cellular Ca2+ regulation is altered in bacteremic rat skeletal muscle, 45Ca2+ uptake was measured in soleus muscles 12 h after an intraperitoneal bacterial (Escherichia coli) injection. Some rats received diltiazem (2.4 mg/kg iv) 10 h after bacterial injection to determine whether calcium blockers could inhibit changes in Ca2+ regulation. Cellular exchangeable Ca2+ was measured in soleus muscles incubated for 5 min to 4 h in Krebs-Ringer bicarbonate (KRB) media (pH 7.4) containing 45Ca2+ (1.5 muCi/ml) and subsequently "washed" in La3+-containing, Ca2+-free KRB media. Bacteremia had no effect on steady-state exchangeable Ca2+, but it significantly reduced the time required to reach half-maximal uptake compared with controls. Diltiazem treatment returned the half-maximal Ca2+ uptake toward control values in bacteremic rat muscles. Depolarization of soleus muscles with high K+ (60 mM) transiently increased Ca2+ uptake in control and bacteremic rat muscles, although the increase was significantly greater (P less than 0.05) in bacteremic rat muscles. The altered skeletal muscle Ca2+ regulation may be due to excessive stimulation of Ca2+ messenger systems, sarcolemmal Ca2+ channels, and/or Ca2+ release from the sarcoplasmic reticulum in response to bacteremia.

Animals↗
PubMed1989

Effect of diltiazem on skeletal muscle 3-O-methylglucose transport in bacteremic rats.

This study examined whether alterations in cellular Ca2+ regulation contribute to previously observed changes in skeletal muscle sugar transport during bacteremia. Fasted male rats received saline (control) or bacteria (4 X 10(10) Escherichia coli/kg) intraperitoneally. Twelve hours later, basal and insulin-mediated 3-O-methylglucose (3MG) transport was measured in isolated soleus muscles. Measurements of 3MG transport in the presence of cytochalasin b or at a low temperature (0.5 degree C) indicated that altered sugar transport in bacteremic rat muscles was not due to nonspecific membrane permeability changes. To determine the role of Ca2+ in the pathogenesis of altered sugar transport during bacteremia, rats were treated with the Ca2+ antagonist diltiazem (DZ, 0.6-2.4 mg/kg) at various times (0, 0 + 7.5, 10 h) after saline or bacterial injection. In bacteremic rats given 2.4 mg/kg DZ at 10 h, basal and insulin-mediated transport were similar to control values. This dose of DZ had little effect on control muscles. The addition of 20 microM DZ to the incubation media did not affect basal or insulin-mediated 3MG transport in bacteremic rat muscles. Addition of the Ca2+ agonist BAY K 8644 to the incubation media had no effect on sugar transport in bacteremic rat muscles but caused alterations in control rat muscles that were comparable to those observed in bacteremia. These results suggest that alterations in Ca2+ regulation could contribute to the previously observed changes in sugar transport in skeletal muscles from bacteremic rats.

3-O-Methylglucose↗
PubMed1989

Alterations in cellular calcium and magnesium during circulatory/septic shock.

Hepatic cellular and mitochondrial Ca2+ and Mg2+ contents and the role of these ions in cellular dysfunction were studied in rat hemorrhagic and endotoxic models which simulate various facets of human circulatory/septic shock states. Hemorrhagic shock was induced by bleeding rats to a state of vascular decompensation at the mean arterial pressure of 40 mm Hg. Endodotoxic shock was ascertained after observing hypoglycemia, hyperlacticacidemia and hemorrhagic bowel lesions in rats injected with Salmonella enteritidis endotoxin (15-20 mg/kg). In a late stage of hemorrhagic shock, significant changes occurred in mitochondrial ion contents; Ca2+ increased and Mg2+ decreased as compared to controls. These ionic changes were correlated with depressed mitochondrial respiratory functions. Addition of 6 mM Mg2+ to shocked-rat isolated mitochondria significantly restored mitochondrial functions. In endotoxic shock, hepatic Mg2+ content was significantly lower compared to controls. Treatment of rats with the calcium antagonist, diltiazem, several minutes prior to endotoxin injections, resulted in restoration of hepatic cellular Mg2+ to control levels. Studies of hepatic intracellular Ca2+ regulation in endotoxic shock revealed (1) an elevation in free cytosolic Ca2+, (2) a blunting of catecholamine-mediated increase in cytosolic Ca2+ and (3) an inhibition of intracellular messenger-induced Ca2+ release by the endoplasmic reticulum.

Animals↗
PubMed1988

Plasma catecholamines during E. coli bacteremia in conscious rats.

Fasted, conscious male rats, prepared with arterial and venous cannulas, were given doses (10(10)-10(11) organisms/kg) of live Escherichia coli bacteria. Heart rate and blood pressure were recorded, and arterial plasma samples were taken preinjection and at 30, 180, and 360 min after bacterial administration. Plasma was analyzed for lactate, glucose, norepinephrine (NE), and epinephrine (E). Rats given E. coli were normotensive but with significant tachycardia (P less than 0.05 vs. saline). Plasma NE and E levels increased severalfold during bacteremia (P less than 0.05 for all comparisons). Rats were euglycemic but had a sixfold increase in lactate 6 h (P less than 0.05) after E. coli treatment. Additional rats were subjected to the same protocol but had been made tolerant to bacterial endotoxin by multiple injections over the course of several days. Endotoxin-tolerant rats were also tolerant to live E. coli administration (P less than 0.05, 24 h survival) and had significantly reduced levels of E and NE at 6 h compared with nontolerant bacteremic rats (P less than 0.05). Increases in heart rate and plasma lactate were not significant in endotoxin-tolerant rats. These results suggest profound sympathetic activation during acute bacteremia with attenuated activation in endotoxin-tolerant rats. Tolerance appears to reduce the afferent stimuli that are presumably activated during the course of bacteremia.

Animals↗