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PubMed1975

Renin secretion from rat renal cortical cell suspensions.

Renin secretion of rat renal cortical cell suspensions was studied as a functionof variations in the composition of the suspension media. Renin secretion was determined by incubating samples of the suspension media with rat renin substrate and measuringthe angiotensin I generated by radioimmunoassay. Increasing the medium sodium concentration (50-144 meq/liter) linearly decreased the rate of renin secretion. This relationship was unaffected by adding ouabain (10 minus 3 M) to the suspensions. The addition of furosemide (from 10 minus 5 to 10 minus 3 M) stimulated renin secretion. Considered with previous observations, these results suggest that renin secretionis controlled by some function of sodium on the lumenal boder of the macula densa cells.

Angiotensin II↗
PubMed1980

Improved renal cortical tubule suspension: spectrophotometric study of O2 delivery.

An improved cortical tubule suspension from the rabbit kidney is described that contains almost entirely proximal convoluted tubules with little contamination by cellular debris or glomeruli. These tubules appear to be capable of active transepithelial transport, since their tubular lumina are open and ouabain inhibits 70% of the QO2. In previous preparations with closed tubular lumina, ouabain inhibited QO2 only 40% and the base-line QO2 was one-half to one-quarter that of the present preparation. The delivery of oxygen to this tubule suspension was compared to that of slices by three different means: oxygen consumption kinetics, ATP content, and, more directly by spectrophotometric monitoring of the redox state of cytochrome oxidase. Results demonstrate that the cortical slice is evidently oxygen deficient even when the bath PO2 is in excess of 570 mmHg. In contrast, the tubule suspension is shown to be adequately oxygenated even at a bath PO2 of 10 mmHg. This tubule suspension will be highly applicable for the analysis of aerobic metabolism in a functionally intact renal preparation by optical, electrode, and biochemical assay technologies.

Adenosine Triphosphate↗
PubMed1985

Bone and muscle atrophy with suspension of the rat.

A modification of the Morey tail suspension model was used to determine atrophic responses of rat bone and muscle with 14-90 days unloading of the hindlimbs. Bone uptake of methylene diphosphonate followed a phasic pattern similar to changes in bone formation rate in immobilized dogs and rats. Increased uptake at 60 days (P = 0.01, femur) indicated an increased bone metabolism. Regional densitometry demonstrated a preferential loss of bone mineral in the trabecular mass (P = 0.02) at 30 days and in the cortical shaft by 90 days (P = 0.03). Maximal muscle atrophy occurred within 14-30 days. The gastrocnemius was less severely affected by suspension than by immobilization techniques, whereas the soleus atrophied (by weight) similarly, suggesting that muscle atrophy in the suspension model is distinctly different from immobilization atrophy. One significant response of skeletal muscle to suspension was an altered blood distribution. Muscle blood distribution changes reflect the hypodynamic state of muscle that continues to contract but probably at an altered rate in response to altered functional demands.

Animals↗
PubMed1986

A new photometric method for oxygen consumption measurements in cell suspensions.

A new technique is described for measuring O2 consumption rates and O2 concentrations in suspensions of respiring cells. Aliquots of a cell suspension kept in a special thermostated precision syringe are injected into the measuring system in defined time intervals. The O2 content of these samples is determined photometrically, as reported previously. The O2 consumption per cellular wet weight and/or per single cell can be calculated from the cell volume fraction, the physical density, the cell concentration in the suspension, and the time-dependent decline of the O2 concentration in the precision syringe. The minimum detectable amount of O2 is 0.1 microliter O2, which corresponds to 0.001 (vol/vol) of O2 if a 100-microliters sample of suspended cells is analyzed. Reproducibility of the O2 consumption measurement is 9% of the measured value. The advantages offered by this method are the straightforward calibration in absolute terms, the short time required for one analysis (2-6 min), a high sensitivity, the simultaneous determination of overall O2 concentration and O2 consumption rates in cell suspensions, and the great variability in the application.

Animals↗
PubMed1987

Interaction of compensatory overload and hindlimb suspension on myosin isoform expression.

The purpose of this study was to investigate the role of chronic weight-bearing activity as the primary inducer of compensatory muscle growth and changes in myosin isoform expression in rodent fast-twitch plantaris muscle. Thus, female rats were subjected to the independent and simultaneous exposure of functional overload (induced via synergist removal) and hindlimb unweighting (suspension) for 6 wk. Groups (n = 7/group) consisted of normal-control (NC); overload (OV); normal-suspension (N-SUS); and overload-suspension (OV-SUS). Body weight of both suspension groups was significantly less than both the NC and OV groups (P less than 0.001). Compared with the NC group, normalized plantaris weight (mg/g body wt) of both the OV and OV-SUS groups was greater, whereas that of the N-SUS was lower (P less than 0.001). However, normalized plantaris weight was greater in OV compared with OV-SUS by 35% (P less than 0.001). Myofibril protein content (mg/g) and Ca2+-regulated myofibril adenosinetriphosphatase (ATPase) specific activity were similar for all groups except that ATPase was lower in the OV group compared with the other groups (P less than 0.05). Native myosin isoform analysis revealed a significant increase in the expression of slow and intermediate myosin and the repression of fast myosin 1 (Fm1) in OV compared with NC. This shift in expression was not as pronounced in the OV-SUS group. Interestingly, only traces of slow myosin were observed in the N-SUS group compared with the other groups. These results suggest that weight bearing is an essential component of the overload model for inducing significant increases in both muscle mass and slow myosin isoform expression. Second, lack of weight bearing, while not markedly affecting fast myosins, appears to repress the expression of slow myosin.

Animals↗
PubMed1987

Time course of soleus muscle myosin expression during hindlimb suspension and recovery.

This study examined the time course of adult rodent soleus muscle myofibril and myosin isoform protein expression after 4, 8, 16, 28, and 56 days of hindlimb unweighting by tail suspension (S). The time course of soleus muscle recovery (R) was also examined after 28 days of hindlimb unweighting with an additional 4, 8, 16, and 28 days of unrestricted cage activity. During suspension, soleus muscle myofibril protein rapidly decreased from 34.3 +/- 3.1 (1.96SE) mg/pair in the control (C) group to 6.9 +/- 1.4 (1.96SE) mg/pair in S (t = 56 days). The calculated first-order degradation rate constant for this loss was kd = 0.17 days-1 [half time (t1/2) = 4.1 days]. The estimated slow myosin (SM) isoform content decreased from 13.4 +/- 2.0 (1.96SE) mg/pair in C to 2.1 +/- 0.2 (1.96SE) mg/pair in S (kd = 0.19 days-1, t1/2 = 3.6 days). The relative proportion of other myosin isoforms was increased at 28 and 56 days of suspension, reflecting an apparent de novo synthesis and the loss of SM. Recovery of contractile protein after 28 days of suspension was slower for both the myofibril protein and the SM isoform (kd = 0.07 days-1, t1/2 = 10 days). These data suggest that loss of weight bearing specifically affected the mechanisms of contractile protein expression reflected in soleus muscle protein degradation processes. In addition, the expression of the myosin isoforms were apparently differentially affected by the loss of weight-bearing activity.

Adrenal Glands↗
PubMed1989

Effect of hindlimb suspension on VO2 max and regional blood flow responses to exercise.

Male rodents were studied before and after undergoing one of three treatment conditions for 9 days: 1) cage control (n = 15, CON), 2) horizontal suspension (n = 15, HOZ), and 3) head-down suspension (n = 18, HDT). Testing included measurements of maximal O2 uptake (VO2 max) and select cardiovascular responses to graded treadmill exercise. VO2 max expressed on an absolute basis (ml/min) was significantly decreased after HOZ (-14.1 +/- 2.5%) and HDT (-14.3 +/- 2.0%), while being essentially unchanged in CON (-1.0 +/- 3.3%). Significant reductions in body weight were observed after both HOZ (-10.1 +/- 4.2 g) and HDT (-22.5 +/- 3.3 g), whereas CON animals exhibited a significant increase in weight (10.4 +/- 3.8 g). As a result, when VO2 max was normalized for body weight, all groups exhibited similar significant reductions of 6-7%. Although no differences in heart rate and blood pressure response to graded exercise were observed, the HDT group exhibited greater increases in mesenteric resistance at the same absolute exercise intensity. Furthermore, both suspended groups had higher iliac resistance values during exercise at similar relative exercise conditions, suggesting that muscle blood flow during treadmill running may have been reduced after suspension. In general, the decrements associated with the HOZ and HDT conditions were similar. It was concluded that reduction in exercise capacity and altered cardiovascular responses to exercise observed after 6-9 days of suspension were attributable to a combination of hypokinesia, lack of hindlimb weight bearing, or restraint, rather than to hydrostatic influences associated with HDT.

Animals↗
PubMed1990

Contractile properties of rat soleus muscle after 15 days of hindlimb suspension.

The properties of the contractile elements interacting to develop force in atrophied rat soleus muscle were studied by using single skinned fibers, which permitted direct access to the contractile apparatus. Muscle atrophy was induced by 15 days of hindlimb suspension. Suspension resulted in a decrease of maximal tension relative to an important decline in fiber diameter. Ca affinity of the contractile proteins was not changed insofar as the tension-pCa relationship was not shifted along the pCa axis. However, after hindlimb suspension 1) the value of the Hill coefficient from the tension-pCa curve was found to be higher, 2) a higher Ca threshold for activation was reported, and 3) a significant increase in contraction kinetics was described. All these results suggested that after suspension the mechanical properties of the slow-twitch soleus appeared to resemble more closely those of a fast-twitch muscle. Our results were in complete agreement with published histochemical data.

Animals↗
PubMed1992

Rat soleus muscle fiber responses to 14 days of spaceflight and hindlimb suspension.

Morphological and enzymatic responses in fibers expressing fast, slow, or both types of myosin heavy chain (MHC) were studied in rats after 14 days of spaceflight (COSMOS 2044) or hindlimb suspension. Although the percentage of slow-twitch fibers was unchanged, a higher percentage of fibers that expressed both slow and fast MHC was observed in flight and suspended rats than in synchronous ground-based controls. The soleus was 25 and 34% smaller than control after 14 days of flight and suspension, with the reduction in fiber cross-sectional area (CSA) being greater in slow- than in fast-twitch fibers in both experimental groups. The activities of succinate dehydrogenase (SDH) and alpha-glycerophosphate dehydrogenase (GPD) were not significantly affected by flight or suspension. The total SDH activity (i.e., SDH activity x CSA) decreased significantly in the slow-twitch fibers of the flight and the fast-twitch fibers of the suspended rats, in large part due to fiber atrophy. A shift in MHC expression in 14 and 9% of the fibers in flight and suspended rats occurred without a change in myosin adenosinetriphosphatase activity. The SDH and GPD activities of the fibers that expressed both slow and fast MHC were slightly higher than the slow-twitch fibers and slightly lower than the fast-twitch fibers. These data indicate that events were initiated within 14 days of spaceflight or suspension that began to reconfigure the protein profiles of 9-14% of the slow-twitch fibers from typical slow-twitch toward those of fast-twitch fibers, while all fibers were dramatically losing total protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases↗
PubMed1994

Influence of suspension on the oxidative burst by rat neutrophils.

The influence of spaceflight on the oxidative burst of neutrophils is not known. The present study was designed to evaluate the influence of antiorthostatic suspension, a ground-based modeling system designed to simulate certain aspects of weightlessness that occur after spaceflight, on the capacity of rat neutrophils to express the oxidative burst, an important host defense mechanism against microbial pathogens. Rats were suspended in whole body harnesses in the antiorthostatic orientation for a 3- or 7-day period. Control rats were suspended orthostatically or allowed to remain in vivarium cages without the attachment of any suspension materials. After suspension, peripheral blood was harvested and neutrophils were isolated by density gradient centrifugation. The enriched neutrophil preparations were stimulated with N-formyl-methionyl-leucine-phenylalanine and phorbol myristic acid to induce the oxidative burst. It was found that neutrophils isolated from suspended animals released the same levels of superoxide anion as did vivarium control animals that were not suspended, indicating that whole body suspension did not alter this aspect of rat neutrophil function.

Animals↗
PubMed1994

Cardiac protein content and synthesis in vivo after voluntary running or head-down suspension.

The adaptive responses of myocardial protein metabolism to chronic increases in work load were evaluated in juvenile female Sprague-Dawley rats. Rats were studied under four conditions: normal weight bearing (N), voluntary wheel running (WR) for < or = 4 wk, head-down-tilt suspension for 7 days (HS), or wheel running (2 or 3 wk) followed by 7 days of suspension (WR-HS). WR activity plateaued after 2 wk at 16 km/day and was maintained through week 4. WR did not affect normal whole body growth. Protein metabolism was studied by measuring heart protein content and in vivo fractional rate of protein synthesis with the [3H]phenylalanine "flooding dose" method. Two weeks of WR increased (P < 0.05) absolute heart protein content (22%) and protein synthesis (21%) relative to age-matched N group values. These differences in protein content and synthesis were maintained for > or = 4 wk. Rats failed to gain significant body weight during suspension. Heart protein content increased (P < 0.05) by 12% to 26% as did protein synthesis (14% to 22%) in HS compared with N group. In WR-HS group, cardiac protein content and protein synthesis were maintained at significantly elevated levels. These findings indicate that 1) high-volume WR by young rats provides a convenient noninvasive method for producing rapid and substantial cardiac hypertrophy, which results, at least in part, from enhanced cardiac protein synthesis; and 2) head-down suspension of sedentary juvenile rats leads to increased cardiac protein synthesis, which helps to increase cardiac protein content despite a lack of whole body growth.

Animals↗
PubMed1998

Granulocyte-macrophage colony-forming-unit assay and liquid suspension culture of bone marrow and peripheral blood in newly diagnosed childhood acute lymphoblastic leukemia.

Granulocyte-macrophage colony-forming-unit (CFU-GM)-assays and/or liquid suspension cultures of bone marrow (BM) and peripheral blood (PB), using human placental conditioned medium as a source of colony-stimulating activity, were performed in 125 children with newly diagnosed acute lymphoblastic leukemia (ALL). Unlike the previous small series which invariably showed poor BM granulocyte-macrophage colony and cluster formations, we found 3 growth patterns. A poor-growth pattern was seen in 101 patients. A low-growth pattern with BM colony and cluster formations in the lower range of the normal limits was observed in 6 patients. All 6 patients had low-risk ALL, and the in vitro growth may reflect the residual normal CFU-GM in BM. Thirdly an over-growth pattern was noted in one patient with refractory ALL. In 49 cases where liquid suspension cultures of BM were performed, 4 had a normal-growth pattern and 42 had apoptotic cells and/or few residual granulocytic/monocytic cells after 7 days of culture. However, in both B cell ALL cases and in 1 refractory T cell ALL case with hyperleukocytosis, repetitive liquid suspension cultures of BM and PB consistently revealed an extensive growth of lymphoblasts. Thus, an over-growth pattern in either the BM CFU-GM assay or liquid suspension culture of BM or PB in children with ALL at diagnosis may indicate a refractory or aggressive disease. A low-growth pattern in the BM CFU-GM assay in children with ALL at diagnosis probably predicts a favorable outcome.

Bone Marrow Cells↗
PubMed2001

Surgery for stress incontinence: a non-randomised trial of colposuspension, needle suspension and anterior colporrhaphy.

OBJECTIVE: To compare the outcome, 12 months after surgery, of three principal categories of procedure (colposuspension, needle suspension and anterior colporrhaphy) used for treating stress incontinence. METHODS: A non-randomised trial design comparing 221 colposuspensions, 54 needle suspensions and 130 anterior colporrhaphies performed by 49 surgeons in 18 hospitals in the North Thames health region in 1993-1994. Four outcomes were considered: complications, severity of stress incontinence, social impact of incontinence, and activities of daily living score. Results were adjusted, using logistic regression, for 13 case-mix variables. RESULTS: Significant differences existed between the three procedures in the characteristics of the patients. The cure rate varied by procedure (colposuspension 34% dry; needle suspensions 13%; anterior colporrhaphy 19%). Two thirds of women, however, reported an improvement (colposuspension 75%; needle suspension 68%; anterior colporrhaphy 55%). After adjusting for confounders, colposuspension was significantly more likely to result in an improvement than anterior colporrhaphy (odds ratio 2.2). While this was reflected in improvements in the social life of the women, the difference between procedures did not reach statistical significance (p = 0.05). CONCLUSION: These results suggest surgery for stress incontinence in typical settings is not as good as reported in textbooks. This is mostly because this study was based on women's own reports (and not surgeons' reports) and partly because of the unselected nature of the patients. The information on outcomes given to patients should be reviewed and a large, pragmatic randomised trial including patient-assessed outcomes is needed.

Female↗
PubMed1984

Endoscopic bladder neck suspension (Stamey-Pereyra) in female urinary stress incontinence. Long-term follow-up of 66 patients.

Of 86 consecutive female patients with urinary stress incontinence treated by endoscopic bladder neck suspension according to the method of Stamey and Pereyra, 56.1% have already had at least one unsuccessful operation. After following up 66 patients (76.7%) for a mean of 4 years (minimum 20 months), 90.9% were free of symptoms or at least markedly improved. The main complications were residual urine of more than 100 cm3 in 3 patients (4.5%) and fistulas around one or both of the suspension sutures in 10 patients (15.2%). 2 of the 3 patients with obstruction remained continent when one of the suspension sutures was removed under local anesthesia. 8 of the 10 in whom one (n = 7) or both (n = 3) suspension sutures were removed because of fistulas remained continent. Long-term results are excellent, even in patients with previous operations.

Adult↗
PubMedDate not supplied

Responses of juxtaglomerular cell suspensions to various stimuli.

Cell suspensions were prepared from rat renal cortical tissue by dispersion with 0.1% collagenase. Unit gravity sedimentation in a 1%-4% Ficoll gradient resulted in a single-cell suspension enriched in juxtaglomerular (JG) cells. Both the cellular renin activity and the amount of renin released into the supernatant increased with time when the suspensions were incubated for 1 hour at 37 degrees C in tissue culture medium. These cells responded to epinephrine and norepinephrine by increasing both synthesis and release of renin. The response was blocked by timolol but not by phenoxybenzamine. Cell suspensions prepared in the same manner but using 0.25% trypsin as the dispersing enzyme neither synthesized nor released renin into the tissue culture medium when similarly incubated. Trypsin-dispersed cells did not respond to catecholamine stimulation. Renin synthesis and release in collagenase-dispersed JG cells were unaltered by changes in Na, K, or Ca ion concentrations. Angiotensin II inhibited release, while saline extracts of clipped kidney from renal hypertensive rats stimulated renin release by these cells.

Angiotensin II↗
PubMed2004

Cryopreserved rat, dog and monkey hepatocytes: measurement of drug metabolizing enzymes in suspensions and cultures.

Metabolism in fresh and cryopreserved (CP) rat, dog and monkey hepatocyte suspensions and cultures was measured using midazolam (CYP3A), tolbutamide (CYP2C), dextromethorphan (CYP2D) and p-nitrophenol (glucuronosyl S-transferases (UGT), sulphotransferases (ST)). CYP3A, CYP2C9, CYP2D6, UGT and ST enzyme functions in fresh and CP rat, dog and monkey hepatocyte suspensions were retained - CP rat hepatocytes lost some CYP2C activity but this was restored by adding NADPH or by placing the cells in culture, suggesting that the enzyme was still functional. Phase 2 activities were equivalent in fresh and CP hepatocyte suspensions. In some cases, incubation conditions increased the rate of metabolism, possibly reflecting de novo cofactor synthesis. However, this effect was substrate and species dependent and was not always the same in fresh and CP cells. CYP3A, CYP2C, CYP2D, UGT and ST activities at 24 hours of culture of rat and monkey hepatocytes were not compromised by cryopreservation. CYP3A, CYP2D but not CYP2C were lower in 24-hour cultures of CP dog hepatocytes than in fresh cells. Despite being lower than fresh cells, UGT activity in dog CP hepatocytes did not decrease from 0 to 24 hours of culture. Species-specific metabolism of p-nitrophenol could be demonstrated in both CP cell suspensions and cultures. In conclusion, these data suggest that the enzyme characteristics of fresh and CP hepatocytes from each species and under specific incubation conditions should be considered when carrying out metabolism studies of new compounds.

Animals↗
PubMed1980

Spontaneous spatio-temporal organization in yeast cell suspension.

Under suitable conditions of substrate supply, cell suspensions of yeast Saccharomyces uvarum may display synchronized oscillations of glycolytic metabolism. Also in an unstirred suspension, spatial patterns can be induced by continuously feeding enzymically liberated glucose into the system. the absorption of NADH in the suspension was measured in an annular cuvette. The observed structures display moving waves, which travel over a distance of some centimetres in the cell suspension and seem to originate from a volume element in which the phase is ahead of those of its neighbours. The wavelength is estimated to be abot 10 cm and the velocity of spreading of the wave is about 5 cm/min. Waves are only observed in the beginning of glycolytic oscillations. Subsequently the spatial synchrony usually increases and destroys the previous organization. Similar spatio-temporal structures are observed during aggregation of the slime mould Dictyostelium discoideum and in the chemical reaction system of Belousov and Zhabotinsky.

Absorption↗
PubMed1981

Differentiation of Dictyostelium discoideum mutant cells in a shaken suspension culture and the effect of cyclic AMP.

In Dictyostelium discoideum, 16 mutants in which cells differentiate into spores and stalk cells without normal morphogenesis were isolated. All these mutants are rapidly developing and capable of differentiating in a shaken suspension of phosphate buffer. The developmental kinetics of specific activities of enzymes in one of the mutants, HTY 1851, cultured in the suspension was compared with that in the parental strain, X2, developed on a solid surface. Most of the enzyme activities appeared much earlier and the peaks of the activities were lower in HTY1851 than X2, but the order or appearance of the activities was the same in both the strains cultured under the conditions described above. These results suggest that the biochemical steps in the development of the mutant in a shaken suspension are essentially the same as those of the parental strain X2 on a solid surface. It was also found that addition of cyclic AMP (2.5 X 10-5 M to 1 X 10-4 M) to the mutant cell suspension 6-8 h after the initiation of development induced an increase in the number of spores and the specific activities of some enzymes to values twice as high as those of an untreated control.

Cell Differentiation↗