Expression of 4-ene steroid 5 alpha-reductase messenger ribonucleic acid in the rat epididymis during postnatal development.

The regulation of epididymal 5 alpha-reductase mRNA is multifactorial and segment-specific. To further investigate the regulation of the message for the enzyme, the expression of 5 alpha-reductase mRNA in the rat epididymis was studied as a function of postnatal development. Developmental changes in 5 alpha-reductase mRNA concentrations were assessed by probing Northern blots with the full-length cDNA for rat steroid 5 alpha-reductase. In the first experiment the effect of postnatal age on 5 alpha-reductase mRNA concentrations in the caput-corpus and cauda epididymides was studied. Male rats, taken at 1-week intervals between the ages of 7-91 days, were used. In both epididymal regions, the mRNA for 5 alpha-reductase was present at all ages examined; it appeared in the immature animal at least 2 weeks before detectable 5 alpha-reductase enzyme activity. In the caput-corpus epididymidis, mRNA levels for 5 alpha-reductase decreased by half between postnatal days 7 and 21, rose 5-fold by day 56, and then remained constant through day 91. No change with postnatal age, however, was observed in the cauda epididymidis. In the second experiment, the longitudinal distribution of 5 alpha-reductase mRNA on postnatal days 21, 42, 49, 56, 77, and 91 was studied. The mRNA levels for 5 alpha-reductase increased remarkably, by 6- to 7-fold, in the initial segment of the caput epididymidis between postnatal days 21 and 42 and stayed constant thereafter. However, no significant change comparable to that found in the initial segment was observed in the adjacent proximal caput region or in any of the other epididymal segments. Thus, the 5-fold rise in 5 alpha-reductase mRNA concentrations that occurred in the caput-corpus epididymidis in the first experiment can be attributed solely to changes in the initial segment. We conclude that steady state concentrations of epididymal 5 alpha-reductase mRNA vary dramatically at different postnatal ages and are highly specific with respect to epididymal segment.


In vivo evidence that inhibin is a gonadotropin surge-inhibiting/attenuating factor.

Hyperstimulation of ovarian function through exogenous gonadotropin administration suppresses the preovulatory surges of both LH and FSH. This phenomenon has been demonstrated to be the result of a non-steroidal factor released from the ovary that has been designated as either gonadotropin surge-inhibiting factor or attenuating factor (GnSIF/AF). To examine the possibility that inhibin might possess the activity ascribed to this factor, endogenous inhibin was immunoneutralized in female rats under conditions known to stimulate GnSIF/AF activity. Inhibin-like immunoreactivity was found to be significantly elevated in FSH-treated rats prior to the time of the gonadotropin surges. Spontaneous preovulatory surges of both LH and FSH were observed in saline-treated (control) rats that were subsequently treated with either anti-inhibin serum (AS) or normal sheep serum (NS). FSH-treatment completely prevented the occurrence of gonadotropin surges in rats subsequently treated with NS. In FSH-injected rats subsequently treated with AS, however, normal preovulatory surges of both LH and FSH were observed. These results indicate that inhibin may be the factor responsible for the suppression of the preovulatory gonadotropin surges in FSH-treated rats and, at least in this species, may be a GnSIF/AF.


Modulation of cytoplasmic calcium signaling in rat pituitary gonadotrophs by estradiol and progesterone.

The stimulatory action of GnRH on gonadotropin secretion from cultured rat pituitary cells is modulated by estradiol (E) and progesterone (P). Since secretory responses to GnRH are initiated by phosphoinositide hydrolysis and Ca2+ mobilization, the effects of gonadal steroids on the pattern of Ca2+ signaling were analyzed in single pituitary gonadotrophs. Increasing concentrations of GnRH elicited a spectrum of [Ca2+]i signals in single gonadotrophs, ranging from subthreshold to threshold-oscillatory and biphasic (spike & plateau) responses. In E-treated gonadotrophs, short-term P treatment shifted subthreshold [Ca2+]i responses to oscillatory and oscillatory to biphasic responses, whereas long-term P treatment shifted oscillatory to subthreshold [Ca2+]i response profiles. These changes parallel the effects of P on GnRH-induced LH release, and indicate that the modulatory effects of ovarian steroids on gonadotropin secretion include a significant action on the Ca2+ signaling pathway.


Structure of human type II 5 alpha-reductase gene.

The best known activity of steroid 5 alpha-reductase is the transformation of testosterone into dihydrotestosterone, the most potent androgen. Two types of human steroid 5 alpha-reductase cDNAs and the type I gene have previously been isolated and characterized. This report describes the isolation and characterization of the human type II 5 alpha-reductase gene, the gene most likely responsible for male pseudohermaphroditism due to 5 alpha-reductase deficiency as well as the one presumed to be involved in a major androgen-related diseases such as prostate cancer and benign prostatic hyperplasia. The type II 5 alpha-reductase gene contains five exons of 352, 164, 102, 151 and 1695 bp, respectively, which share 43.8% to 64.1% homology with exons of the corresponding type I gene. These exons are separated by four introns of greater than 29, and approximately 2.3, 2.0 and 3.0 kb. Analysis of primer extension products by polyacrylamide gel electrophoresis as well as by subcloning and sequencing reveals a start site located 71 nucleotides upstream the ATG initiating codon.


The effect of hypothyroidism on Sertoli cell proliferation and differentiation and hormone levels during testicular development in the rat.

In this study we show that 6-propyl-2-thiouracil (PTU) treatment of Wistar rats from birth up to day 26 p.p. retards the morphological differentiation of Sertoli cells, and prolongs the proliferation of these cells up to day 30. Sertoli cell numbers per testis, determined at day 36, were increased by 84% compared to controls. PTU treatment increased serum thyroid-stimulating hormone (TSH) levels and reduced serum levels of thyroxine (T4) from 5 days onwards, indicative of severe hypothyroidism. Follicle-stimulating hormone (FSH) levels were reduced from day 5 to 9, normal at day 12 and 16, and reduced again from day 20 to 36. Inhibin levels were decreased from day 9 to 20 and increased at 36 days of age. The increase in the number of Sertoli cells per testis in PTU treated rats, as has been reported in the present study, is likely to be responsible for the increased testis size observed by other groups (1) in these animals, when adult.


Biliary stent occlusion--a problem solved with self-expanding metal stents? European Wallstent Study Group.

The main limitation in the endoscopic palliation of malignant biliary obstruction is due to stent blockage. One of the factors thought to be of importance is the diameter of the endoprosthesis. In this paper, we report the results of a multicenter European study with a one cm diameter self-expanding metal stent (Wallstent) in 103 patients with malignant biliary obstruction. Insertion of the stent following guidewire positioning was successful in 97.1% of the patients without any cases of de novo cholangitis developing after the endoscopic procedure. The median follow-up for the entire group was 145 days. In all but 3 patients, the stent expanded to more than 80% of its maximum diameter. Two patients had ongoing cholangitis after stent insertion. Long-term complications manifested by late cholangitis, were seen in 18% of the cases after a median interval of 125 days. The occlusion rate by biliary sludge was 5% after a median time period of 175 days which is substantially less than the 21% occlusion rate reported for polyethylene stents. In conclusion, our results show that the Wallstent can be easily placed in distal and mid-CBD strictures after guidewire passage, with most of the patients having a- good drainage effect. The occlusion rate by biliary sludge is significantly less than for conventional polyethylene stents, but the occlusion by tumor ingrowth is substantial. A disadvantage is the high cost of the Wallstent. Further randomized trials will be required to determine the cost-benefit ratio for the use of this stent.


Technical failure of biliary metal stent deployment in a series of 116 applications.

Biliary metal stents are thought to offer improved long-term palliation of malignant biliary obstruction due to a lower incidence of migration and clogging. Placement of these stents is technically more complicated than that of plastic endoprostheses and requires two experienced physicians. We report the incidence and reasons for apparent malfunction of expandable metal stent deployment (Wallstents and Strecker stents). In 116 applications of 82 Wallstents (endoscopic approach: n = 33, transhepatic approach: n = 49), we observed 19 cases of stent malfunction due to technical problems of stent delivery. In 13 cases (15.8%), the restraining membrane of the Wallstent could not be retracted sufficiently to deliver the stent. There were 6 (17.6%) failures in 34 cases of Strecker stent deployment. In 3 cases, we noted difficult balloon removal, including avulsion of the balloon catheter shaft within the endoscope during attempted balloon removal in one case. In one case, the Strecker stent could only be released partially, requiring subsequent endoscopic extraction. In two patients, only partial expansion of one end of the Strecker stent could be achieved. Given the significant malfunction rate of expandable metal stents during stent delivery, further improvements in the delivery system of the metal stents are required.


Three years experience with endoscopic variceal ligation for treatment of bleeding varices.

Endoscopic variceal ligation (EVL) was developed as an alternative to endoscopic variceal sclerosis (ES) because of the high complication rate seen with ES. The new technique involves placement of small elastic bands around the variceal channels in the distal esophagus. The first 146 consecutive patients treated with EVL during the period from August, 1986 to July, 1989 are reported. Portal hypertension was caused by alcoholic liver disease in 93 of these patients. The average age of the patients was 53 years and 66% were males. All of the patients had recently bled from esophageal varices. At the time of treatment, 23% of the patients were actively bleeding. They were all treated acutely with EVL and had repeated treatments with the long-term goal of variceal eradication. The overall survival was 73%. Varices were eradicated or reduced to grade one in 78% of the 125 patients who were followed for more than 30 days. Variceal eradication required a mean of 5.5 sessions. Recurrent bleeding occurred in 44% of the total patient population. There were no major complications from EVL. It is concluded from this non-randomized experience that EVL is an effective treatment for bleeding esophageal varices and that it appears to be as effective as sclerotherapy with fewer complications.


Silicone-covered expandable metallic stents in the esophagus: an experimental study.

Modified silicone-covered Gianturco expandable metallic stents were placed in the normal esophagus of six young pigs. Following endoscopic examination, the stents were placed using endoscopic and fluoroscopic control. The animals were observed for eating behavior and weight gain. Stents appeared to be tolerated well based on these parameters. Three stents remained in position for the full study period, and three stents migrated into the stomach during the study. The pigs were sacrificed at four weeks and postmortem examination performed. Esophageal wall thickening and nodular inflammation were noted at the sites where the wire skirts penetrated the mucosa. Injury was limited to the region of the wire skirts and there was no injury due to the radial force of the stent body. There was no free perforation. This preliminary study suggests that endoscopic and fluoroscopic placement of modified silicone-covered Gianturco stents in the esophagus is feasible and safe. More extensive animal studies, followed by clinical investigation for palliation of malignant strictures, are warranted.


Early recurrence of obstructive jaundice after placement of a self-expanding metal endoprosthesis.

Self-expanding metal endoprostheses have ben used in the treatment of malignant obstructive jaundice for a few years. We report on a patient with obstructive jaundice due to a metastasis of a squamous cell lung cancer into the pancreatic head who received an expandable metal endoprosthesis and suffered recurrent jaundice and cholangitis after 38 days. On repeat ERCP, a very narrow stenosis was seen in the stent lumen caused by tumor ingrowth through the mesh. Intraluminal biopsies showed squamous cell carcinoma. A 10 Fr plastic endoprosthesis was inserted through the self-expanding stent with good drainage. On the basis of this experience, we feel that when faced with a patient with obstructive jaundice due to metastatic disease from a rapidly progressive carcinoma, the use of the currently available self-expanding metal endoprosthesis should be discouraged until the results of prospective studies with large numbers of patients have proven its superiority over the plastic endoprostheses. Meanwhile, modifications to prevent tumor ingrowth through the mesh of the stent are under investigation.


Haemorrhage due to erosion of a metal biliary stent through the duodenal wall.

Self-expanding, metal biliary stents have recently been used in malignant obstructive jaundice as their large diameter reduces the likelihood of occlusion by biliary sludge and bacterial biofilm. However, there is a significant rate of late obstruction by tumour overgrowth and infiltration through the wire mesh. Our case of stent erosion through the duodenal wall, resulting in massive haemorrhage, is a hitherto unreported serious complication of the Wallstent (Schneider). Its occurrence may be suggested by continuous upper abdominal pain, gastrointestinal bleeding or late expansion of the stent.


Endoscopic application of Gianturco-Rösch biliary Z-stent.

Ginaturco-Rösch Z-stent is a new self-expanding stainless steel stent for drainage of the obstructed biliary system. It has been used only with a percutaneous transhepatic technique. We describe a new method for endoscopic retrograde placement of a Gianturco-Rösch biliary Z-stent. This report describes our experience for endoscopic application of a Gianturco-Rösch biliary Z-stent in a patient with a benign stricture and multiple common bile duct stones.


Acute pancreatitis complicating the insertion of a self-expandable biliary metal stent.

A patient is reported who developed acute pancreatitis complicated by pseudocyst formation immediately after the insertion of a self-expanding metallic biliary stent via the percutaneous route. The distal end of the stent was positioned into the duodenum. Endoscopists should be aware of this potential complication of metallic biliary stents.


The analysis of human serum transferrins with the PhastSystem: quantitation of microheterogeneity.

The PhastSystem was used for three different analyses of human serum transferrins (Tf) by isoelectric focusing: (i) The distribution of iron over the two iron binding sites by separation of the four forms: apoTf, TfFeC, FeNTf and Fe2Tf by using less than 1 microL serum or plasma. The focused bands were visualized by immunoprecipitation and quantified by densitometry. (ii) The relative proportions of the diferic sialo-transferrin fractions in human sera or plasma were determined in a similar way. (iii) The presumed genetic variants can be determined. The methods are easy and reproducible and require much less sample (0.3 microL), time (1 h per run) and antibody (10 microL/sample) than crossed immunoelectrofocusing.


Analysis of carbohydrate-mediated heterogeneity and characterization of N-linked oligosaccharides of glycoproteins by high performance capillary electrophoresis.

Capillary zone electrophoresis (CZE) has been investigated as an alternative method to analyze the carbohydrate moieties of glycoproteins. Carbohydrate-mediated microheterogeneity of the recombinant plasminogen activator (rt-PA) was examined. The glycoprotein was resolved in multiple electrophoretic species using CZE but the separation was complicated by adsorption of the molecules to the wall of the capillary. The influence of several parameters, such as pH, molarity of the buffer and addition of a cationic additive, on the separation of glycopeptides was investigated. High resolution and reproducible separations of rt-PA glycopeptides carrying hybrid and complex type chains were obtained using either a 100 mM phosphate buffer, pH 6.6, or a 100 mM Tricine buffer, pH 8.2, containing 1.25 mM of putrescine. N-Oligosaccharides from fetuin, t-PA and alpha 1-acid glycoprotein were separated within 20 min on the basis of both their sialic acid content and their structure. The use of an oligosaccharide fingerprinting technique, such as the present one, could have many applications in biotechnology to assess, for example, the consistency of production of a glycoprotein or for analytical glycoprotein chemistry.


Improvement of BF typing and of BF F subtyping after neuraminidase treatment in the unconverted and converted factor B.

When neuraminidase-treated sera are analyzed by agarose gel isoelectric focusing, the factor B (BF) banding pattern is reduced to predominantly one major band without cathodically positioned bands. This not only makes unequivocal typing of BF allotypes possible but also the reliable distinction of all BF F subtype phenotypes with delimitation of "BF F subtype variants". With this new method, serum aging affects the BF determination to a lesser extent than when applying methods that separate native sera. We show that sialylation is not responsible for the BF F subtype polymorphism. All of the investigated BF allotype bands, including those characteristic of the subtypes, show functional hemolytic activity. The banding pattern after removal of neuraminic acid residues ranges from pH 6.8 to 7.3 for factor B, from pH 5.3 to 5.9 for the Ba fragment, and from pH 8.2 to 8.7 for the Bb fragment. The protein structure of factor B is also discussed. Eliminating the superimposition of bands in different BF allotypes, as demonstrated by these methods, proved to be necessary for the detection of hypomorphic BF gene products (BF QL), which are expressed by assumed BF*Q0 alleles in heterozygous genotypes. This allows investigation of BF*Q0 alleles on a protein level, which complements molecular genetic approaches.


Analysis of collagen type III by uninterrupted sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting: changes in collagen type III polymorphism in aging rats.

A new method of type III collagen analysis by uninterrupted sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) combined with immunoblotting was developed. The electrophoresis was carried out with gels containing 4 M urea. A negatively charged reducing agent, thioglycolic acid, was added to the running buffer of the cathodic reservoir between 15 and 20 min after Bromphenol Blue (BPB) migrated to the top of the separating gel, to reduce interchain disulfide binding of the collagen. The polymorphic type III collagens, i.e., an alpha-chain derived from a trimer [alpha 1(III)]3 with interchain disulfide bonds but without covalent cross-links, alpha 1(III), a beta-chain with covalent cross-links, beta(III), or an alpha-chain released from a trimer without reduction of the disulfide bonds, alpha*1(III), were identified by immunostaining and quantified by densitometry. Using this method, changes in collagen type III polymorphism with aging were examined in the aorta, brachial artery, and skin of rats. The total quantity of collagen type III decreased with aging in all tissues. beta(III) was the major component in the aorta and brachial artery, but alpha 1(III) was the major component in the skin. With increasing age from 3 to 60 weeks, the ratio of beta(III) to alpha 1(III), which is correlated with the extent of covalent cross-linking, showed a steep increase in the aorta but only a slight increase in the skin and it remained almost constant in the brachial artery.(ABSTRACT TRUNCATED AT 250 WORDS)


A new concept for sequencing DNA by capillary electrophoresis.

It is proposed that the scaling symmetry of constant charge density with increasing molecular weight, which prevents the separation by electrophoresis of DNA molecules in solution (with respect to molecular weight) be broken by the attachment of a perturbing entity (protein, virus or charged sphere) to one end of the molecule. An application of this idea to a concept for sequencing DNA by capillary electrophoresis is discussed, and the possibility of using the reattachment of the RecA protein to separate large segments of DNA in solution by electrophoresis following sequence-specific cleavage is mentioned.


A new technique for detecting oxytocinase activity in electrophoresis gels.

A sensitive staining method for the detection of oxytocinase (EC 3.4.11.3) activity in electrophoresis gels has been described. The method is based on the enzymatic release of p-nitroaniline (PNA) from two specific synthetic oxytocinase substrates, S-benzyl-L-cysteine-p-nitroanilide (BCN) and L-leucine-p-nitroanilide (LN), respectively. The PNA is then diazotized with sodium nitrite and subsequently coupled to a chromogen, N-(1-naphthyl)-ethylenediamine dihydrochloride (NED) to produce a deep pink/magenta colored azo-dye at the site of oxytocinase activity.


Tissue-specific pattern of nonhistone high mobility group proteins in various organs of the chicken.

Electrophoretic analysis of tissue-specific differences of nonhistone high mobility group (HMG) proteins from nuclei of various organs of the chicken revealed that in organs with a higher proportion of replicating cells (thymus, Bursa Fabricii, spleen) the relative amount of HMG-17 is considerably higher than that of HMG-14; however, in transcriptionally active organs with a very small proportion of replicating cells (glandular stomach, liver) HMG-14 and HMG-17 are present at roughly equal and low amounts. In glandular stomach, liver and spleen, the relative contents of both HMG-1 and HMG-2 are markedly lower than in thymus and Bursa Fabricii. Moreover, the total amount of HMG proteins is higher in those organs which contain replicating lymphocytes.


Separation of proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of soluble, aqueous polymers: Ficoll and polyvinylpyrrolidone.

In previous studies we have demonstrated that water soluble polymers of dextran and methylcellulose, when incorporated into sodium dodecyl sulfate (SDS)-polyacrylamide gels, can be used to sharpen the protein bands. We have extended these studies to enhance the separations in the mid-molecular weight range. Enhanced separation of protein molecular weight markers between 30,000-67,000 was best achieved in SDS-gels containing 2.5% polyvinylpyrrolidone (average molecular weight 10,000).


Anthropometric, muscle strength, and spinal mobility characteristics as predictors in the rating of acceptable loads in parcel sorting.

The rating of acceptable load (RAL) attained with a standard test (RALSt) and a wrk-simulating test (RALW) for postal parcel sorting was related to anthropometric, muscle strength, and spinal mobility characteristics of 18 male sorters. The subjects comprised a subsample of 103 experienced male sorters who carried out the RAL tests at postal sorting centres. The dynamic hand-grip endurance correlated significantly (p = 0.036) to the RALSt results. Correspondingly, there was a significant correlation (p = 0.044) between the ratio of maximal isometric strength of trunk extension to body weight and the RALW. The dynamic hand-grip endurance predicted 26% of the variation in the RALSt; in the RALW the maximal isometric strength of trunk flexion to body weight ratio predicted 24%. The subjects who rated heavier weights for RALSt tended to have a better trunk mobility. The dynamic endurance of hand-grip muscles, trunk strength, and spinal flexibility seemed to be the most powerful predictors for the psychophysically assessed 'acceptable loads' in experienced workers performing manual materials handling tasks.


Logistical and ergonomic transportation capacity for refuse collection workers: a work physiology field study.

In a work physiology field study, the work flow and the electrocardiogram were recorded throughout whole shifts for six male refuse (garbage) collection workers who transported and emptied 1.1 m3 refuse containers. The work rate (WR), indicated by the number of 1.1 m3 containers emptied per unit of time, and the work pulse rate (WPR) were determined in the data evaluation. The work pulse rate increases with the work rate. The functional relationship can be approximated by a linear regression function (WPR = 20.9 + 35.8.WR). The work pulse rate reaches such high values that it must be assumed that the work cannot be performed continuously throughout the whole working day. Consequently, regular breaks should be provided. When determining the necessary duration of the breaks, it was assumed that an equilibrium between fatigue and recovery should be maintained during the working day. The recovery breaks should be at least long enough for the heart rate to return to the resting level. The necessary duration of the breaks was determined on the basis of the present field study and the laboratory investigations described in the literature. A minimum duration of 10 min per working hour results from the calculations. The breaks should be taken regularly at about hourly intervals. A transportation-capacity model is provided for the planning of the deployment of refuse workers. It permits calculation of the number of 1.1 m3 containers a three-man crew, comprising the refuse collection truck driver and two loaders, can empty during one shift. Both the logistical and ergonomic transportation capacities can be determined using the model. The logistical transportation capacity (TCL) indicates the number of containers which can be emptied per shift by a crew when only criteria such as the optimal utilization of the working time are considered. The ergonomic transportation capacity (TCE) is understood as the number of containers which can be emptied per shift taking into consideration the workers' strain and the recovery breaks they require. TCE amounts to between about 140 and 160 containers per shift depending on the type of city district. TCL is higher by between 20 and 30 containers per shift. In order to meet the demand of protecting workers' health, the number of containers to be emptied per shift by a three-man crew should not exceed the ergonomic transportation capacity.


The metabolic cost of backpack and shoulder load carriage.

Eleven healthy male volunteer soldiers (mean [SD] age 24.0 [2.8] years, stature 174.1 [5.2] cm, body weight 73.2 [10.8] kg, body fat 14.2 [5.0]% and maximal oxygen uptake 4.1 [0.4] 1 min-1) walked at 4.8 km h-1 on a motor driven treadmill for 5 min at each of three gradients (0, 2.5 and 5%) whilst carrying a two-part 26 kg load either on each shoulder or strapped to a backpack frame. The load was made up of two cylinders, one weighing 18.4 kg and the other weighing 7.6 kg. For all treadmill gradients the mean (SD) backpacking heart rates and oxygen uptakes (0% gradient, 122 [10] beats min-1, 1.51 [0.11] 1 min-1; 2.5% gradient, 135 [10] beats min-1, 1.81 [0.13] 1 min-1; 5% gradient, 155 [7] beats min-1, 2.21 [0.11] 1 min-1) were significantly (p less than 0.001) lower than for shoulder load carriage (0% gradient, 130 [9] beats min-1, 1.70 [0.12] 1 min-1, 2.5% gradient, 147 [8] beats min-1; 2.01 [0.10] 1 min-1; 5% gradient 164 [9] beats min-1, 2.39 [0.11] 1 min-1). The relative oxygen cost of backpacking was 4.3-4.7% VO2 max lower than for shoulder load carriage. It is concluded that the metabolic cost of backpacking an asymmetric two part 26 kg load was significantly less than for shoulder load carriage when walking at 4.8 km h-1 on a treadmill over gradients of 0-5%.(ABSTRACT TRUNCATED AT 250 WORDS)


Evaluation of impact loading in overhead work using a bolt pistol support.

This paper evaluates the effects of a bolt gun support stand on impact loading of the hand-arm system while firing spiked bolts into a steel beam. Ten healthy male subjects fired two bolts into the centre of the beam, one with and one without the aid of the support stand. During firing of a bolt gun, the shock sequences were recorded by two accelerometers. The analysis showed that peak values of acceleration decreased more than eightfold during firing with the bolt pistol support. The shock damping effect due to the bolt support was noted at the thumb (metacarpal level) and elbow bones.


Physical training: a tool for increasing work tolerance limits of employees engaged in manual handling tasks.

The lack of physical fitness is a contributing factor to the etiology of musculoskeletal disorders resulting from the manual handling of material in industry. Thus the major objectives of this paper were (1) to discuss the role of physical fitness in the control of occupational injuries; (2) to review techniques available in the exercise physiology literature for increasing human physical capacity in industry; and (3) to review and evaluate studies on the effects of physical training on individuals engaged in manual handling tasks.


Effects of job-simulated flexibility and strength-flexibility training protocols on maintenance employees engaged in manual handling operations.

This study examined the effects of four flexibility and strength-flexibility training protocols on the dynamic strength, endurance time, and truncal flexibility of 24 maintenance employees engaged in manual material handling operations. The study was conducted over an 8-week period. Significant improvement in physical capacity was obtained through flexibility training either by a progressive increase in the holding time with fixed exercise repetition or a progressive increase repetition with fixed holding time. The flexibility and strength-flexibility training protocols exhibited similar effects on physical capacity. It was suggested that a follow-up flexibility protocol should be performed on a daily basis in order to maintain the gains obtained in flexibility during the intensive/short training programme.


Understanding and evaluating manual handling injuries: NIOSH research studies.

This paper presents an overview of NIOSH research aimed at characterizing and identifying intervention strategies for reducing musculoskeletal injuries during manual handling activities. Surveillance and evaluative research projects are reviewed. Future research directions of the Institute are also discussed.


Ergonomic workplace assessment in a health care context.

Nurses, together with other groups in the caring professions, show a high incidence of musculoskeletal injuries as a consequence of the handling of equipment and patients. This is an internationally recognized consequence of the work in these professions, and studies worldwide have investigated these problems. The UK Health and Safety Executive, as well as the European Community (EC) Directives, require workplaces to be assessed for their hazardousness. Ergonomics is the recognized discipline in relation to assessing whether the work, equipment, or environment match the capacities of the people concerned. An assessment procedure is presented, together with proposals for its use, which can form the basis for the ergonomic assessment of workplaces in health care, and give guidance for their improvement.


A novel and conserved salt-induced protein is an important determinant of salt tolerance in yeast.

We have isolated a novel yeast gene, HAL1, which upon overexpression improves growth under salt stress. In addition, disruption of this gene decreases salt tolerance. Therefore HAL1 constitutes a rate-limiting determinant for halotolerance. It encodes a polar protein of 32 kDa located in the yeast cytoplasm and unrelated to sequences in data banks. The expression of this gene is increased by high concentrations of either NaCl, KCl or sorbitol. On the other hand, the growth advantage obtained by overexpression of HAL1 is specific for NaCl stress. In cells overexpressing HAL1, sodium toxicity seems to be counteracted by an increased accumulation of potassium. The HAL1 protein could interact with the transport systems which determine intracellular K+ homeostasis. The HAL1 gene and encoded protein are conserved in plants, being induced in these organisms by salt stress and abscisic acid. These results suggest that yeast serves as a convenient model system for the molecular biology of plant salt tolerance.


Three-dimensional crystal structure of recombinant murine interferon-beta.

The crystal structure of recombinant murine interferon-beta (IFN-beta) has been solved by the multiple isomorphous replacement method and refined to an R-factor of 20.5% against 2.6 A X-ray diffraction data. The structure shows a variant of the alpha-helix bundle with a new chain-folding topology, which seems to represent a basic structural framework of all the IFN-alpha and IFN-beta molecules belonging to the type I family. Functionally important segments of the polypeptide chain, as implied through numerous gene manipulation studies carried out so far, are spatially clustered indicating the binding site(s) to the receptor(s). Comparison of the present structure with those of other alpha-helical cytokine proteins, including porcine growth hormone, interleukin 2 and interferon gamma, indicated either a topological similarity in chain folding or a similar spatial arrangement of the alpha-helices.


Sequences directing dihydrolipoamide dehydrogenase (E3) binding are located on the 2-oxoglutarate dehydrogenase (E1) component of the mammalian 2-oxoglutarate dehydrogenase multienzyme complex.

Sequences located in the N-terminal region of the high M(r) 2-oxoglutarate dehydrogenase (E1) enzyme of the mammalian 2-oxoglutarate dehydrogenase multienzyme complex (OGDC) exhibit significant similarity with corresponding sequences from the lipoyl domains of the dihydrolipoamide acetyltransferase (E2) and protein X components of eukaryotic pyruvate dehydrogenase complexes (PDCs). Two additional features of this region of E1 resemble lipoyl domains: (i) it is readily released by trypsin, generating a small N-terminal peptide with an apparent M(r) value of 10,000 and a large stable 100,000 M(r) fragment (E1') and (ii) it is highly immunogenic, inducing the bulk of the antibody response to intact E1. This 'lipoyl-like' domain lacks a functional lipoamide group. Selective but extensive degradation of E1 with proteinase Arg C or specific conversion of E1 to E1' with trypsin both cause loss of overall OGDC function although the E1' fragment retains full catalytic activity. Removal of this small N-terminal peptide promotes the dissociation of dihydrolipoamide dehydrogenase (E3) from the E2 core assembly and also affects the stability of E1 interaction. Thus, structural roles which are mediated by a specific gene product, protein X in PDC and possibly also the E2 subunit, are performed by similar structural elements located on the E1 enzyme of the OGDC.


Transmembrane helical interactions: zeta chain dimerization and functional association with the T cell antigen receptor.

Members of the zeta family of receptor subunits (zeta, eta and gamma) are structurally related proteins found as components of the T cell antigen receptor (TCR) and certain Fc receptors. These proteins share the ability to form disulfide-linked dimers with themselves and with other members of the family. Comparison of the amino acid sequences of zeta and gamma reveals a significant degree of homology, which is highest within their membrane-spanning domains. Analysis of their transmembrane sequences on a helical wheel projection suggests that all of the identical amino acids are clustered on one face of a potential alpha-helix. This face contains the only cysteine residue within zeta, suggesting that this conserved region may function to mediate dimerization. Indeed, replacing the transmembrane domain of the Tac antigen (alpha chain of the interleukin-2 receptor) by that of the zeta chain resulted in the formation of disulfide-linked dimers of Tac. The conserved aspartic acid residue found in the zeta and gamma transmembrane sequences was found to play a role in disulfide linkage. Replacing the aspartic acid with a lysine but not with an alanine or valine residue allowed formation of disulfide-linked dimers. The ability of the aspartic acid residue to support dimerization was dependent upon its position within the helix. Thus, these observations indicate that residues within the zeta transmembrane domain play a critical role in the formation of disulfide-linked dimers. Expression of zeta mutants in zeta-deficient T cells revealed that the zeta transmembrane domain is also responsible for reconstituting transport of functional TCR complexes to the cell surface and differentiated the requirements for disulfide-linked dimerization per se from assembly of the TCR complex.


The human poly(ADP-ribose) polymerase nuclear localization signal is a bipartite element functionally separate from DNA binding and catalytic activity.

Poly(ADP-ribose) polymerase (PARP, EC 2.4.2.30) is a zinc finger DNA-binding protein involved in DNA repair processes in eukaryotes. By deletion and extensive site-directed mutagenesis, its DNA-binding domain fused to the N-terminus of beta-galactosidase was shown to contain a nuclear localization signal (NLS) of the form KRK-X(11)-KKKSKK (residues 207-226). In vitro, both the DNA-binding capacity and the polymerizing activity of PARP are independent of the nuclear location function. Each basic cluster is essential but not sufficient on its own for this function, while both motifs together are. Crucial basic amino acids (K207, R208 and K222) in each of these two motifs are required for nuclear homing. The results presented here support the concept that the human PARP NLS is an autonomous functional element and belongs to the class of bipartite NLSs. We show that the linear distance between the two basic clusters is not crucial. Insertional mutation analysis leading to a partial reversion of the cytoplasmic phenotype displayed by the mutant K222I highlights the crucial positioning of this lysine. The structure-function relationship of the second cluster of basic residues is discussed.


A novel gene, spp91-1, suppresses the splicing defect and the pre-mRNA nuclear export in the prp9-1 mutant.

Processing and export of nuclear pre-mRNA are believed to be competing processes in the nucleus. In order to identify factors which are involved in these processes, we isolated suppressors that relieve the growth defect of a prp9-1 temperature-sensitive mutant strain of Saccharomyces cerevisiae. The prp9-1 mutation was previously shown to abolish splicing and to target pre-mRNA to the cytoplasm. One of the suppressors, spp91-1, corrects the prp9-1 growth defect through partial restoration of splicing and by a complete reversion of the pre-mRNA escape phenotype. This suppressor is specific for two prp9 alleles and cannot substitute for PRP9 function. The mutant and wild-type alleles of SPP91 were cloned and sequenced. SPP91 encodes a novel protein essential for mitotic growth whose sequence contains motifs indicative of a nuclear localization. In vivo depletion of SPP91 in a prp9-1 genetic background is lethal and is associated with reduced amounts of spliced mRNA and accumulation of pre-mRNA. This observation strongly supports the hypothesis that SPP91 encodes a PRP factor. We suggest that spp91-1 increases pre-mRNA retention in the nucleus by improving the formation of the spliceosome and thereby allowing a larger proportion of the pre-mRNA molecules to be spliced.


Histone H3 N-terminal mutations allow hyperactivation of the yeast GAL1 gene in vivo.

Recent work has shown that the yeast histone H4 N-terminus, while not essential for viability, is required for repression of the silent mating loci and activation of GAL1 and PHO5 promoters. Because histone H3 shares many structural features with histone H4 and is intimately associated with H4 in the assembled nucleosome, we asked whether H3 has similar functions. While the basic N-terminal domain of H3 is found to be non-essential (deletion of residues 4-40 of this 135 amino acid protein allows viability), its removal has only a minor effect on mating. Surprisingly, both deletions (of residues 4-15) and acetylation site substitutions (at residues 9, 14 and 18) within the N-terminus of H3 allow hyperactivation of the GAL1 promoter as well as a number of other GAL4-regulated genes including GAL2, GAL7 and GAL10. To a limited extent glucose repression is also alleviated by H3 N-terminal deletions. Expression of another inducible promoter, PHO5, is shown to be relatively unaffected. We conclude that the H3 and H4 N-termini have different functions in both the repression of the silent mating loci and in the regulation of GAL1.


Hold back of RNA polymerase II at the transcription start site mediates down-regulation of c-myc in vivo.

Premature termination of transcription is assumed to be an important mechanism of c-myc regulation. Induction of terminal differentiation in the promyelocytic leukemia cell line HL60 by dimethyl-sulfoxide (DMSO) is accompanied by a block of RNA elongation within the first exon of the c-myc gene. We have studied the 3'-structure of incompletely elongated transcripts in (i) nuclear RNA of induced and uninduced HL60 cells, (ii) nuclear run-on RNA, and (iii) RNA of in vitro transcribed c-myc constructs. Elongation of c-myc RNA stopped in all three transcriptional systems at similar sites distributed 150-350 bases downstream of the P2 promoter. When HL60 cells were induced to terminal differentiation the short c-myc exon 1 specific RNAs disappeared in nuclear RNA. This implied that RNA polymerase II (pol II) does not continue to transcribe c-myc exon 1 in induced HL60 cells as suggested by earlier nuclear run-on experiments. Therefore, kinetic experiments with small oligonucleotides as probes were performed to determine the start position of pol II on c-myc exon 1 in nuclear run-ons. The results demonstrate that all RNA polymerases are localized at the c-myc P2 promoter in DMSO-treated HL60 cells. Preparation of nuclei for run-on experiments induces a release of pol II from the c-myc P2 promoter leading to the strong nuclear run-on signal on c-myc exon 1. Thus, down-regulation of c-myc in differentiating HL60 cells occurs by retention of pol II at the transcription start site.


Homeotic control in Drosophila; the scabrous gene is an in vivo target of Ultrabithorax proteins.

The regulatory functions of transcription factors encoded by the Ultrabithorax (Ubx) gene initiate genetic programmes essential for segmental identity and morphogenesis in Drosophila. Based on the formation of DNA-protein adducts in intact nuclei and immunoselection procedure, we cloned genomic targets for Ubx proteins. One clone was studied in detail. It encompasses parts of the last intron and exon of the scabrous (sca) gene, which encodes a secreted protein involved in cellular communication during neurogenesis. Five motifs, presenting the ATTA core, which is shared by most homeodomain binding sites, were found in the nucleotide sequence of this clone. We detail here the dynamic pattern of sca transcript accumulation during embryogenesis and show that mutation of Ubx results in the ectopic transcription of sca in the first abdominal segment. We propose that a direct interaction of Ubx with cis-acting elements in sca negatively regulates the gene. Transcript localization in several combinations of deficiencies in the Bithorax complex (BX-C) indicates that sca is downregulated by abdominal A (abdA) and Abdominal B (AbdB), and suggests that it is a common target of the three genes of BX-C.


The extramacrochaetae gene provides information for sensory organ patterning.

The Drosophila adult epidermis displays a stereotyped pattern of bristles and other types of sensory organs (SOs). Its generation requires the proneural achaete (ac) and scute (sc) genes. In the imaginal wing disc, the anlage for most of the thoracic and wing epidermis, their products accumulate in groups of cells, the proneural clusters, whose distribution prefigures the adult pattern of SOs. These proteins then induce the emergence of SO mother cells (SMCs). Here, we show that the extramacrochaetae (emc) gene, an antagonist of the proneural function, is another agent that contributes to SO positioning. In the wing disc, emc is expressed in a complex and evolving pattern. SMCs appear not only within proneural clusters but also within minima of emc expression. When one of these spatial restrictions is eliminated, by ubiquitously expressing ac-sc, SMCs still emerge within minima of emc. When in addition, the other spatial restriction is reduced by decreasing emc expression, many ectopic SMCs emerge in a relatively even spaced and less constant pattern. Thus, the heterogeneous distribution of the emc product is one of the elements that define the positions where SMCs arise. emc probably refines SMC (and SO) positioning by reducing both the size of proneural clusters and the number of cells within clusters that can become SMCs.


TAR-independent transactivation by Tat in cells derived from the CNS: a novel mechanism of HIV-1 gene regulation.

The Tat protein of human immunodeficiency virus type 1 (HIV-1) is essential for productive infection and is a potential target for antiviral therapy. Tat, a potent activator of HIV-1 gene expression, serves to greatly increase the rate of transcription directed by the viral promoter. This induction, which seems to be an important component in the progression of acquired immune deficiency syndrome (AIDS), may be due to increased transcriptional initiation, increased transcriptional elongation, or a combination of these processes. Much attention has been focused on the interaction of Tat with a specific RNA target termed TAR (transactivation responsive) which is present in the leader sequence of all HIV-1 mRNAs. This interaction is believed to be an important component of the mechanism of transactivation. In this report we demonstrate that in certain CNS-derived cells Tat is capable of activating HIV-1 through a TAR-independent pathway. A Tat-responsive element is found upstream within the viral promoter that in glial-derived cell lines allows transactivation in the absence of TAR. Deletion mapping and hybrid promoter constructs demonstrate that the newly identified Tat-responsive element corresponds to a sequence within the viral long terminal repeat (LTR) previously identified as the HIV-1 enhancer, or NF-kappa B domain. DNA band-shift analysis reveals NF-kappa B binding activity in glial cells that differs from that present in T lymphoid cells. Further, we observe that TAR-deleted mutants of HIV-1 demonstrate normal late gene expression in glial cells as evidenced by syncytia formation and production of viral p24 antigen.(ABSTRACT TRUNCATED AT 250 WORDS)


Cell cycle regulation of glucocorticoid receptor function.

Glucocorticoid receptor (GR) nuclear translocation, transactivation and phosphorylation were examined during the cell cycle in mouse L cell fibroblasts. Glucocorticoid-dependent transactivation of the mouse mammary tumor virus promoter was observed in G0 and S phase synchronized L cells, but not in G2 synchronized cells. G2 effects were selective on the glucocorticoid hormone signal transduction pathway, since glucocorticoid but not heavy metal induction of the endogenous Metallothionein-1 gene was also impaired in G2 synchronized cells. GRs that translocate to the nucleus of G2 synchronized cells in response to dexamethasone treatment were not efficiently retained there and redistributed to the cytoplasmic compartment. In contrast, GRs bound by the glucocorticoid antagonist RU486 were efficiently retained within nuclei of G2 synchronized cells. Inefficient nuclear retention was observed for both dexamethasone- and RU486-bound GRs in L cells that actively progress through G2 following release from an S phase arrest. Finally, site-specific alterations in GR phosphorylation were observed in G2 synchronized cells suggesting that cell cycle regulation of specific protein kinases and phosphatases could influence nuclear retention, recycling and transactivation activity of the GR.


Cloning and functional characterization of the rat stomach fundus serotonin receptor.

A DNA segment homologous to the third exons of the serotonin 1C and 2 receptor genes was isolated from a mouse genomic library. The positions of the introns flanking these exons were conserved in the three genes. To examine whether the new fragment was part of an active gene, we used a quantitative PCR protocol to analyse rat RNAs from different tissues and ages. The gene was expressed in stomach fundus at an abundance of 1 x 10(5) mRNA molecules. This tissue contracts in response to serotonin via a receptor that has previously resisted classification. We constructed a cDNA library from rat stomach fundus and isolated clones containing 2020 bp inserts with open reading frames of 465 amino acids comprising seven putative membrane-spanning regions. The protein was transiently expressed in COS cells and binding of serotonergic ligands to the membranes was analysed. The pharmacological profile resembled that described for the serotonin-stimulated contraction of the stomach fundus. After expression of this receptor in Xenopus oocytes, the application of serotonin triggered the typical chloride current which presumably results from the activation of phospholipase C. The coupling to this response system was less efficient than that of the 5-HT1C or 5-HT2 receptors.


Assessment of the in vivo aneuploidy/micronucleus assay in mouse bone marrow cells with 16 chemicals.

Assessment of the in vivo aneuploidy/micronucleus Assay in bone marrow cells with 16 chemicals is described. This assay is based on the detection of kinetochores (KC) in micronuclei (MN) by antikinetochore-specific (CREST) antibodies. Among sixteen chemicals tested, six were known clastogens, three were known aneuploidy-inducers, and the other seven were suspected spindle poisons. These chemicals were tested for their ability to induce micronuclei with kinetochore(s) in bone marrow cells of CD-1 mice. The majority of MN formed in bone marrow cells treated with aneuploidy-inducing agents contained kinetochore(s) which are considered to be formed from whole chromosomes or centric fragments, while in clastogen treated bone marrow cells, majority of them contained no kinetochore(s) which are considered to be formed from acentric chromosomal fragments. Classification of chemicals into either aneuploidy inducing agents or clastogens is based on the relative frequency of MN with and without KC, respectively. These results suggest that the in vivo aneuploidy/micronucleus assay has a great potential to identify aneuploidy-inducing agents.


Cytogenetic effects of vincristine sulfate and ethylene dibromide in human peripheral lymphocytes: micronucleus analysis.

Micronuclei kinetics and persistence in mononucleated and binucleated human peripheral lymphocytes following short-term (4 hr) and continuous (until harvest) in vitro exposure to vincristine sulfate (VS) and ethylene dibromide (EDB) were studied. Lymphocytes were exposed to chemicals for various doses and harvested at different culture times. Micronucleus frequencies were scored in both mononucleated and binucleated cells on the same slide. VS-treated cells showed a significantly higher incidence of micronucleus in both mononucleated and binucleated cells than controls (P less than 0.01). The cells treated continuously with VS produced comparatively higher frequencies of micronucleated cells than those treated for 4 hr. Highest micronuclei frequencies were observed 24 hr after chemical treatment in both mononucleated and binucleated cells and decreased later with time. However, the micronucleus frequencies remained significantly higher than the controls even in the cells harvested at 144 hr. VS induced a large number of micronucleated cells with multiple micronuclei. VS also caused a severe decrease in nuclear division due to cytotoxic effect. Lymphocytes treated with EDB for 4 hr and continuously showed a statistically higher incidence of micronuclei in binucleated cells compared to the controls (P less than 0.05), whereas in mononucleated cells higher micronucleus frequencies were observed only in cultures treated continuously. Continuous presence of EDB induced both dose- and time-dependent increase of micronuclei in both mono- and binucleated cells (P less than 0.05). EDB induced relatively few multiple micronucleated cells in comparison with VS. EDB did not affect nuclear divisions even with continuous treatment. High micronucleus frequencies observed at 144 hr harvest following 4 hr treatment of both EDB and VS suggest the persistence of DNA damage in cells. These studies suggest that micronuclei kinetics in human peripheral lymphocytes depends on the genotoxic potentially and cytotoxicity of a genotoxicant.


Comparison of rat liver parenchymal and nonparenchymal cells in the activation of promutagens.

While the liver consists of both parenchymal cells (PC) and nonparenchymal cells (NPC), virtually all studies on promutagen activation have been performed using PC. To evaluate the comparative roles of PC and NPC in promutagen activation, we cocultivated a cell line generally considered to have an insignificant level of xenobiotic metabolism, Chinese hamster ovary (CHO) cells, with either PC, NPC, or a combination of both. The mixed culture was treated with two promutagens: dimethylnitrosamine (DMN) and 3-methylcholanthrene (3-MC). The induction of 6-thioguanine resistant mutants was evaluated using the well-established CHO/hypoxanthine-guanine phosphoribosyl transferase (HGPRT) assay. Activation of promutagens, as indicated by an increase in mutant frequency in CHO cells, was observed only when the PC were present with the CHO cells during the treatment period. No activation was observed with NPC. Coculturing of PC and NPC yielded essentially the same results as PC alone. P-450 mixed function monooxygenase activity measured by the 7-ethoxycoumarin-O-deethylase assay further substantiates that PC had a significantly higher xenobiotic metabolism activity than NPC. Our study therefore indicates that PC, not NPC, are the major cell population in the liver responsible for the activation of promutagens.


Structure-activity relationship of genotoxic polycyclic aromatic nitro compounds: further evidence for the importance of hydrophobicity and molecular orbital energies in genetic toxicity.

A quantitative structure-activity relationship (QSAR) has been formulated for 15 polycyclic aromatic nitro compounds acting on E. coli PQ37. Upon damage of DNA by these substances beta-galactosidase is induced and can be easily assayed colorimetrically, hence, this is a short-term test for mutagenicity. The QSAR (log SOSIP = 1.07 log P - 1.57 epsilon LUMO - 6.41) is strikingly similar to that found earlier with nitroaromatics acting in the Ames test (TA100) and differs significantly for that found using TA98 organisms. The QSAR brings out in a unique manner the underlying similarity in the two test systems.


Human HPRT mutant database: software for data entry and retrieval.

We have developed a computer database containing information on over 1,000 human hypoxanthine guanine phosphoribosyl transferase (HPRT) mutants. Both published and unpublished data are present. The database itself is maintained in a dBASE format (.DBF) and we provide a set of programs to examine and extract information from the database. A program to input information into the database is also supplied. The database and programs are available directly from us or via remote FTP (file transfer protocol) using BITNET/INTERNET. All programs require an IBM-compatible computer, the MS-DOS operating system (version 3.3 or greater), and a hard disk with about 5 megabytes of free disk space. The purpose of the database is 1) to allow investigators to contribute their HPRT mutants directly to the database in a standardized fashion, and 2) to allow access to the entire database with a set of programs that allows manipulation and extraction of data. For example, using our programs it is possible to i) order the database by base pair position, ii) examine only information regarding mutagenesis by a particular agent, iii) search for a particular author, iv) create a report which contains selected portions of the database, the report can be printed or saved as a file. The database will be updated every several months and distributed.


Mutation spectrum of spontaneous frameshift revertants in yeast using double-strand gap repair.

A mutation spectrum was constructed from a series of randomly isolated spontaneous His+ revertants of the frameshift mutant his4-38 in Saccharomyces cerevisiae. For each true revertant, a 438 bp region encompassing his4-38 on chromosome III was recovered into a shuttle vector by double-strand gap repair. Of the 45 independent His+ revertants sequenced, 44 were -1 base deletions and one revertant was a +2 base insertion. The -1 deletions exhibited a bimodal distribution. Of the bases encompassing the his4-38 region from +153-181, approximately 45% were not involved in a reversion event, although a -1 frameshift within this region will result in a viable His+ revertant. Approximately 49% of -1 events occurred within runs of 3 repeated bases. At these sites the strand-slippage model for frameshift mutation is supported. However, the -1 events occurring at sites of 2 repeated bases and the low frequency (2%) of +2 base insertions suggest that the transiently misaligned template model is a significant mechanism in reversion of his4-38. When the distribution of -1 events at repeated bases was discounted, a hotspot involving a -T at position +163 was resolved.


DNA sequence analysis of spontaneous and N-ethyl-N-nitrosourea-induced hprt mutations arising in vivo in cynomolgus monkey T-lymphocytes.

The study of hprt mutations in cynomolgus monkey T-lymphocytes is part of our effort to understand the mechanisms of mutagenesis in vivo. This primate model allows us to study mutations and their kinetics at the molecular level under well-controlled conditions using recently developed techniques for selection of mutant T-cells and polymerase chain reaction (PCR) amplification of hprt cDNA, which is directly sequenced. This is the first report of the sequence of the coding region of the cynomolgus monkey hprt gene and PCR/DNA sequence analysis of seven spontaneous mutant T-cell clones, as well as 23 mutant clones isolated 63 and 601 days after treatment with ethylnitrosourea (ENU, 77 mg/kg, intraperitoneal). cDNA was reverse transcribed from hprt mRNA directly from a lysate of about 2-4 x 10(3) cells, and a 700 bp fragment including the coding region was amplified by PCR and sequenced. Of the seven spontaneous mutants, only one point mutation (GC----AT transition) was detected, and the other six failed to amplify by PCR, possibly due to functional deletions. Of the 14 mutant clones isolated 63 days after ENU treatment, nine base substitutions were detected in ten clones: four transitions (three AT----GC and one GC----AT) and five transversions (four AT----TA and one AT----CG). Of the nine mutants isolated 601 days after ENU treatment, six single base substitutions were detected in six clones (five AT----TA and one AT----CG transversions), and one mutant had a large deletion or insertion. No changes were detected in three clones (one Day 63 and two Day 601 clones). In summary, only one of 15 single base substitutions isolated after ENU treatment was a GC----AT transition mutation and the rest were transitions and transversions at AT sites.


Muscle sound and electromyogram spectrum analysis during exhausting contractions in man.

The changes in the soundmyogram (SMG) and electromyogram (EMG) frequency content during exhausting contractions at 20%, 40%, 60% and 80% of the maximal voluntary contraction (MVC) were investigated by the spectral analysis of the SMG and EMG detected from the biceps brachii muscles of 13 healthy men. The root mean squares (rms) of the two signals were also calculated. Throughout contraction the EMG rms always increased while this was true only at 20% MVC for the SMG. A marked decrease was detected at 60% and 80% MVC. With fatigue the EMG spectra presented a compression towards the lower frequencies at all exercise intensities. The SMG showed a more complex behaviour with a transient increase in its frequency content, followed by a continuous compression of the spectra, at 60% and 80% MVC, and a nearly stable frequency content at lower contraction intensities. This study suggested that different aspects of the changes in the motor unit's activation strategy at different levels of exhausting contractions can be monitored by SMG and EMG signals.


Effect of carnitine loading on long-chain fatty acid oxidation, maximal exercise capacity, and nitrogen balance.

Carnitine has a potential effect on exercise capacity due to its role in the transport of long-chain fatty acids into the mitochondria for beta-oxidation, the export of acyl-coenzyme A compounds from mitochondria and the activation of branched-chain amino acid oxidation in the muscle. We studied the effect of carnitine supplementation on palmitate oxidation, maximal exercise capacity and nitrogen balance in rats. Daily carnitine supplementation (500 mg.kg-1 body mass for 6 weeks) was given to 30 rats, 15 of which were on an otherwise carnitine-free diet (group I) and 15 pair-fed with a conventional pellet diet (group II). A control group (group III, n = 6) was fed ad libitum the pellet diet. Palmitate oxidation was measured by collecting 14CO2 after an intraperitoneal injection of [1-14C]palmitate and exercise capacity by swimming to exhaustion. After carnitine supplementation carnitine concentrations in serum were supranormal [group I, total 150.8 (SD 48.5), free 78.9 (SD 18.4); group II, total 170.9 (SD 27.9), free 115.8 (SD 24.6) mumol.l-1] and liver carnitine concentrations were normal in both groups [group I, total 1.6 (SD 0.3), free 1.2 (SD 0.2); group II, total 1.3 (SD 0.3), free 0.9 (SD 0.2) mumol.g-1 dry mass]. In muscle carnitine concentrations were normal in group I [total 3.8 (SD 1.2), free 3.2 (SD 1.0) mumol.g-1 dry mass] and increased in group II [total 6.6 (SD 0.5), free 4.9 (SD 0.9) mumol.g-1 dry mass].(ABSTRACT TRUNCATED AT 250 WORDS)


The effect of a high carbohydrate diet on running performance during a 30-km treadmill time trial.

The purpose of the present study was to examine the influence of a high carbohydrate diet on running performances during a 30-km treadmill time trial. Eighteen runners (12 men and 6 women) took part in this study and completed a 30-km time trial on a level treadmill without modifying their food intake (trial 1). The runners were then randomly assigned to a control or a carbohydrate (CHO) group. The CHO group supplemented their normal diets with additional carbohydrate in the form of confectionery products during the 7 days before trial 2; the control group matched the increased energy intake of the CHO group by consuming additional fat and protein. The mean (SEM) carbohydrate intake of both groups was 334 (22) g before trial 1, after which the CHO group consumed 566 (29) g.day-1 for the first 3 days and 452 (26) g.day-1 for the remaining 4 days of recovery. Although there was no overall difference between the performance times for the two groups during trial 2, the CHO group ran faster during the last 5 km of trial 2 than during trial 1 [3.64 (0.24) m.s-1 vs 3.44 (0.26) m.s-1; P less than 0.05]. Furthermore, the 6 men in the CHO group ran the 30 km faster after carbohydrate loading [131.0 (5.4) min vs 127.4 (4.9) min; P less than 0.05], whereas there was no such improvement in times of the men in the control group.(ABSTRACT TRUNCATED AT 250 WORDS)


The effects of a reduced exercise duration taper programme on performance and muscle enzymes of endurance cyclists.

The influence of tapering on the metabolic and performance parameters in endurance cyclists was investigated. Cyclists (n = 25) trained 5 days.week-1, 60 min.day-1, at 75-85% maximal oxygen consumption (VO2max) for 8 weeks and were then randomly assigned to a taper group: 4D (4 days; n = 7), 8D (8 days; n = 6), CON (control, 4 days rest; n = 6), NOTAPER (non-taper, continued training; n = 6). Muscle biopsy specimens taken before and after training and tapering were analysed for carnitine palmityltransferase (CPT), citrate synthase, beta-hydroxyacyl CoA dehydrogenase (HOAD), cytochrome oxidase (CYTOX), lactate dehydrogenase, glycogen and protein. Significant increases in VO2max (6%), a 60-min endurance cycle test (34.5%), oxidative enzymes (77-178%), glycogen (35%) and protein (34%) occurred following training. After the taper, HOAD and CPT decreased 25% (P less than 0.05) and 26% respectively, in the CON. Post-taper CYTOX values were different (P less than 0.05) for 4D and 8D compared with CON. Muscle glycogen levels were increased (P less than 0.05) after tapering in the 4D, 8D and CON, but decreased in NOTAPER. Similarly, power output at ventilation threshold was significantly increased in the 4D (27.4 W) and 8D (27 W) groups, but decreased (22 W) in the NOTAPER. These findings suggest that tapering elicited a physiological adaptation by altering oxidative enzymes and muscle glycogen levels. Such an adaptation may influence endurance cycling during a laboratory performance test.


Exercise-induced hypoxemia in athletes: role of inadequate hyperventilation.

These experiments examined the exercise-induced changes in pulmonary gas exchange in elite endurance athletes and tested the hypothesis that an inadequate hyperventilatory response might explain the large intersubject variability in arterial partial pressure of oxygen (PaO2) during heavy exercise in this population. Twelve highly trained endurance cyclists [maximum oxygen consumption (VO2max) range = 65-77 ml.kg-1.min-1] performed a normoxic graded exercise test on a cycle ergometer to VO2max at sea level. During incremental exercise at VO2max, 5 of the 12 subjects had ideal alveolar to arterial PO2 gradients (PA-aO2) of above 5 kPa (range 5-5.7) and a decline from resting PaO2 (delta PaO2) 2.4 kPa or above (range 2.4-2.7). In contrast, 4 subjects had a maximal exercise PA-aO2 of 4.0-4.3 kPa with delta PaO2 of 0.4-1.3 kPa while the remaining 3 subjects had PA-aO2 of 4.3-5 kPa with delta PaO2 between 1.7 and 2.0 kPa. The correlation between PAO2 and PaO2 at VO2max was 0.17. Further, the correlation between the ratio of ventilation to oxygen consumption vs PaO2 and arterial partial pressure of carbon dioxide vs PaO2 at VO2max was 0.17 and 0.34, respectively. These experiments demonstrate that heavy exercise results in significantly compromised pulmonary gas exchange in approximately 40% of the elite endurance athletes studied. These data do not support the hypothesis that the principal mechanism to explain this gas exchange failure is an inadequate hyperventilatory response.


Pain, thermal sensation and cooling rates of hands while touching cold materials.

Hand cooling and resulting comfort and pain were studied in 12 subjects, while touching six different materials (polyurethane foam, wood, nylon, rustproof steel, aluminium, and temperature-controlled metal) which were initially at ambient temperature. This was done for three ambient temperatures (-10 degrees, 0 degree and 10 degrees C), after pre-exposure exercise or rest, with bare hands or while wearing gloves. The observed cooling curves were analysed as Newtonian cooling curves. The observed time constants appeared to be significantly related to the materials' contact coefficients, the presence of hand protection, the preceding activity, and the interaction between contact coefficient and the presence of hand protection. These parameters also allowed a good description of the time constant (r2 = 0.8) of the related cooling curves. Thermal and pain sensation could be described in terms of the local skin temperature, ambient temperature and hand protection. Equal pain and thermal levels were associated with lower temperatures of the back of the hand than of the contact side. The slightly painful condition was associated with a skin temperature of 16 degrees C for the back and 19 degrees C for the palm of the hand. The pain level appeared to be inversely related to cooling speed. Skin freezing occurred at higher skin temperatures when touching cold objects than when exposed to cold air as a result of reduced supercooling. The regression equations determined allowed calculations to be made of safety limits for hand cooling while in contact with a wide range of materials.


One-handed load carrying--cardiovascular, muscular and subjective indices of endurance and fatigue.

A group of 5 women and 11 men walked on a treadmill, each carrying a weight in the right hand. In separate experiments, the mass was varied to give total exhaustion within 3 min, 5 min, 9 min, and 13 min. In additional experiments 50% and 25% of the masses leading to exhaustion after 5 min were used, and these were stopped after 16 min. Heart rate (fc) and systolic blood pressure (BPs) were measured noninvasively. There was a consistent increase in fc x BPs during the experiments leading to exhaustion, while steady-states were obtained in the nonexhausting trials. An electromyogram (EMG) was recorded with cutaneous electrodes over the flexor carpi ulnaris and flexor digitorum superficialis muscles and the number of zero crossings (ZC) of the EMG signal per time unit were analysed. As the subjects approached exhaustion, the number of ZC declined exponentially, reaching approximately 50% of their initial values. In the nonexhausting experiments, however, the decline was slower and less marked, and during the second half of the experiment the number of ZC increased again. Subjectively, endurance was underestimated by all the subjects. It was concluded that cardiovascular and muscle criteria of fatigue in carrying coincided. Prolonged carrying in one hand of more than 6 kg or 10 kg for young healthy women and men respectively should not be recommended, since it could lead to cardiovascular non steady-states and EMG signs of fatigue.


Simulation of hand cooling due to touching cold materials.

A simple analytical model has been developed to simulate the cooling of the hands due to touching various types of cold material. The model consisted of a slab of tissue, covered on both sides with skin. The only active mechanism was the skin blood flow. The blood flow was controlled by body core temperature, mean skin temperature, and local hand temperature. The blood flowed along the palm before returning via the back of the hand. The control function was adapted from an earlier study, dealing with feet, but enhanced with a cold induced vasodilatation term. The palm of the hand was touching materials that were specified by conductivity and heat capacity. The hand was initially at a steady-state in a neutral environment and then suddenly grabbed the material. The resulting cooling curves have been compared to data from an experiment including six materials (foam, wood, nylon, steel, aluminium and metal at a constant temperature), three temperatures (-10, 0, and 10 degrees C), two thermal states of the body (neutral and 0.4 degrees C raised), and with and without gloves. There was a fair general agreement between the model and the experiment but the model failed to predict three specific effects: the unequal effect of equal 10 degrees C steps in cold surface temperature on the temperature of the palm of the hand, the cooling effect of nylon, and the rapid drop in back of the hand temperature. Nevertheless the overall regression was 0.88 with a standard deviation between model and experiment of about 2.5 degrees C.


Glucose and free fatty acid utilization during prolonged exercise in prepubertal boys in relation to catecholamine responses.

Ten prepubertal boys performed 60-min cycle exercise at about 60% of their maximal oxygen uptake as previously measured. To measure packed cell volume, plasma glucose, free fatty acids (FFA), glycerol and catecholamines, blood samples were drawn at rest using a heparinized catheter and at the 15th, 30th and 60th min of the exercise and after 30 min of recovery. At rest, the blood glucose concentrations were at the lowest values for normal. Exercise induced a small decrease of blood glucose which was combined with an abrupt increase of the noradrenaline concentration during the first 15 min. The FFA and glycerol concentrations increased throughout the exercise linearly with that of adrenaline. Compared to adults, the FFA uptake expressed per minute and per litre of oxygen uptake was greater in children. These results suggested that it is difficult for children to maintain a constant blood glucose concentration and that prolonged exercise provided a real stimulus to hypoglycaemia. An immediate and large increase in noradrenaline concentration during exercise and a greater utilization of FFA was probably used by children to prevent hypoglycaemia.


Cardiovascular responses in paraplegic subjects during arm exercise.

The purpose of this study was to examine cardiovascular responses during arm exercise in paraplegics compared to a well-matched control group. A group of 11 male paraplegics (P) with complete spinal cord-lesions between T6 and T12 and 11 male control subjects (C), matched for physical activity, sport participation and age performed maximal arm-cranking exercise and submaximal exercise at 20%, 40% and 60% of the maximal load for each individual. Cardiac output (Qc) was determined by the CO2 rebreathing method. Maximal oxygen uptake was significantly lower and maximal heart rate (fc) was significantly higher in P compared to C. At the same oxygen uptakes no significant differences were observed in Qc between P and C; however, stroke volume (SV) was significantly lower and fc significantly higher in P than in C. The lower SV in P could be explained by an impaired redistribution of blood and, therefore, a reduced ventricular filling pressure, due to pooling of venous blood caused by inactivity of the skeletal muscle pump in the legs and lack of sympathetic vasoconstriction below the lesion. In conclusion, in P maximal performance appears to have been limited by a smaller active muscle mass and a lower SV despite the higher fc,max. During submaximal exercise, however, this lower SV was compensated for by a higher fc and, thus at the same submaximal oxygen uptake, Qc was similar to that in the control group.


Peak power output predicts maximal oxygen uptake and performance time in trained cyclists.

The purposes of this study were firstly to determine the relationship between the peak power output (Wpeak) and maximal oxygen uptake (VO2max) attained during a laboratory cycling test to exhaustion, and secondly to assess the relationship between Wpeak and times in a 20-km cycling trial. One hundred trained cyclists (54 men, 46 women) participated in the first part of this investigation. Each cyclist performed a minimum of one maximal test during which Wmax and VO2max were determined. For the second part of the study 19 cyclists completed a maximal test for the determination of Wpeak, and also a 20-km cycling time trial. Highly significant relationships were obtained between Wpeak and VO2max (r = 0.97, P less than 0.0001) and between Wpeak and 20-km cycle time (r = -0.91, P less than 0.001). Thus, Wpeak explained 94% of the variance in measured VO2max and 82% of the variability in cycle time over 20 km. We concluded that for trained cyclists, the VO2max can be accurately predicted from Wpeak, and that Wpeak is a valid predictor of 20-km cycle time.


Heart rate overshoot at the beginning of muscle exercise.

A characteristic notch in the heart rate (fc) on-response at the beginning of square-wave exercise is described in 7 very fit marathon runners and 12 sedentary young men, during cycle tests at 30% and 60% of maximal oxygen consumption (VO2max). The fc notch revealed a fc overshoot with respect to the fc values predicted from exponential beat-by-beat fitted models. While at 30% of VO2max all subjects showed a fc overshoot, at 60% of VO2max it occurred in the marathon runners but not in the sedentary subjects. The mean time of occurrence of the fc overshoot from the onset of the exercise was 16.7 (SD 4.7) s and 12.2 (SD 3.2) s at 30% of VO2max in the runners and the sedentary subjects respectively, and 23.8 (SD 8.8) s at 60% of VO2max in the runners. The amplitude of the overshoot, with respect to rest, was 41 (SD 12) beats.min-1 and 31 (SD 4) beats.min-1 at 30% of VO2max in the runners and the sedentary subjects respectively, and 46 (SD 19) beats.min-1 at 60% of VO2max in the runners. The existence and the amplitude of the fc overshoot may have been related to central command and muscle heart reflex mechanisms and thus may have been indicators of changes in the balance between sympathetic and parasympathetic activity occurring in fit and unfit subjects.


Psycho-sensorimotor performance in divers exposed to six and seven atmospheres absolute of compressed air.

In the present study, we investigated the psycho-sensorimotor abilities of divers exposed to 6 ATA (50 m of sea water; corresponding to legal limit for occupational diving in North America, United Kingdom, and Northern Europe), or 7 ATA (60 m; the legal limit in France and Southern Europe) of compressed air (1 ATA = 100,000 Pa), using psychometric tests of manual dexterity, visual choice reaction time, and number ordination. The results of the present study showed that abilities in these tests were not significantly altered by pressure exposure to 6 ATA of compressed air. However, data obtained at 7 ATA showed slight but significant decreases in performance. Nevertheless, a few subjects presented large decreases in performance ranging from -20% to -25% of control. Finally, our results supported the ergonomic point of view that the laws limiting occupational diving to 6 ATA (50 m) are better adapted to reality and the requirements of underwater activity.


D-lactate concentrations in blood, urine and sweat before and after exercise.

The purpose of this study was to investigate changes in the concentrations of D-lactate, L-lactate, pyruvate and methylglyoxal (MG) in body fluids after exercise. Eight untrained male students and five male students who were boat club members engaged in the exercise. Each subject performed runs of short and long duration. Compared to pre-exercise values plasma concentrations of D-lactate, L-lactate and pyruvate increased after running; in trained men by 3.6, 5.0, 3.4 times after short runs and by 1.5, 4.6, 2.0 times after long runs, and in untrained men by 3.0, 12.0, 1.6 times after short runs and 2.5, 5.6, 1.6 times after long runs, respectively. In all cases, the increase of L-lactate was always higher than that of D-lactate after running. The MG contents in red blood cells decreased markedly after running, especially in the untrained students. After short runs the MG concentration had decreased to 13% in the untrained men and 30% in the trained men, and after long runs the concentration had decreased to 41% in the untrained and 60% in the trained men. The MG in plasma and red blood cells appeared to have been utilized during relatively anaerobic exercise, especially by the untrained subjects. The D-lactate and related substances were also determined in urine, but the concentration of these substances showed no relationship to exercise. The D-lactate concentration in sweat samples tripled after short periods of running but the relative concentration to sodium ion concentration was not altered.(ABSTRACT TRUNCATED AT 250 WORDS)


Venous responses to rhythmic exercise in contralateral forearm and calf.

Ten normal healthy subjects performed a rhythmic handgrip at 30% MVC (maximal voluntary contraction) with and without arterial occlusion of the same limb. Contralateral forearm and calf venous capacitance were simultaneously measured by venous occlusion plethysmography. During rhythmic handgrip at 30% MVC contralateral venous capacitance decreased by -7.17% in the forearm and by -5.14% in the calf. With arterial occlusion the decreases in venous capacitance were even more pronounced: contralateral forearm -14.4% and calf -13.1%. In a second set of experiments (n = 5) rhythmic handgrip at 30% MVC with arrest of the forearm circulation 5 s prior to the cessation of contraction was applied to examine the influence of chemically sensitive metaboreceptors per se on the evoked limb venoconstriction. During the postexercise arterial occlusion forearm venous volume decreased further to -30.6% whereas calf venous volume increased slightly but remained below the control value. After the cessation of the arterial occlusion both forearm and calf capacitance returned to baseline values. Thus, this study provided evidence that as well as a chemically generated reflex arising from the working muscle, central command was found to be involved in the increase in venomotor tone in the nonexercising limbs during rhythmic handgrip at 30% MVC.


Stress Doppler echocardiography using dobutamine in coronary patients with and without ischaemia induction.

To examine the effects of dobutamine on pulsed-Doppler left ventricular ejection dynamics and its utility for evaluation of coronary disease (CAD) we studied 10 patients with normal coronaries (Group 1) and 24 patients with significant CAD (greater than or equal to 70% diameter stenosis) using a graded stress infusion (5 to 20 micrograms.kg-1.min-1). Two-dimensional echocardiography was performed to detect regional asynergy and analysed using an 11-segment model. Patients with CAD were divided into those with (Group 3, n = 14) and without (Group 2, n = 10) inducible ischaemia; six patients had reversible involvement of greater than or equal to three segments (subset 3A). Groups were well matched for baseline left ventricular function and all studies were carried out while the patients were not taking cardioactive therapy. Hyperkinetic wall motion was typical and exaggerated hyperkinesis of normal segments was commonly seen in those with baseline (n = 9) or reversible asynergy (n = 14). Normals and CAD patients showed comparable changes in heart rate and blood pressure (P = NS between groups).(ABSTRACT TRUNCATED AT 250 WORDS)


Improved monitoring of myocardial ischaemia during major vascular surgery using transoesophageal echocardiography.

As regional wall motion abnormality (RWA) is the first sign of ischaemia, transoesophageal echocardiography was evaluated as a monitoring device in 51 patients undergoing abdominal aortic surgery. Wall motion of the entire left ventricular wall (nine segments) was semiquantitatively evaluated 15 min before and after aortic cross-clamping and 3 h after declamping. In addition, limb lead II of the electrocardiogram was simultaneously recorded. At baseline, RWA was present in 16 patients (31%). New or worsened RWA 15 min after aortic cross-clamping was seen in 17 patients, of whom 11 had persistent RWA, i.e. it was still present 3 h after declamping. This was associated in seven patients with enzymatically documented myocardial infarction. Only one infarct patient demonstrated ST segment changes of more than 1 mm. Thus, a single electrocardiographic surface lead is insensitive for perioperative myocardial ischaemia detection. Furthermore, new and/or worsened RWA after aortic cross-clamping, which persists until 3 h after declamping, is, to a considerable degree, associated with perioperative infarction.


Non-invasive assessment of mitral stenosis before and after percutaneous balloon mitral valvotomy by Doppler continuity equation.

The continuity equation was used to estimate non-invasively the stenotic mitral valve area by comparison with two other echocardiographic methods (planimetry and pressure half-time) and with Gorlin's formula as the gold standard. The accuracy of the equation of continuity was determined before and 24 h after valvuloplasty in a study group of 21 patients with severe mitral stenosis. According to the equation of continuity, mitral valve area was calculated by the product of the cross-sectional area and the aortic or pulmonary annulus and the ratio of the time velocity integral of the aortic or pulmonary flow to that of the mitral stenotic jet. In pre-valvotomy basal conditions, the Doppler continuity equation demonstrated significant correlations with 2D planimetry (r = 0.72, P less than 0.01), with the pressure half-time method (r = 0.62, P less than 0.01) and with the Gorlin formula (r = 0.66, P less than 0.01). There was no significant difference between the haemodynamic data and the echocardiographic measurements. Twenty-four hours after valvotomy, the Doppler continuity equation also demonstrated significant correlations with 2D planimetry (r = 0.83, P less than 0.01), with pressure half-time (r = 0.82, P less than 0.01) and with the Gorlin formula (r = 0.69, P less than 0.01). However, the haemodynamic measurements significantly overestimated (P less than 0.01) the echographic measurements. Thus, we conclude that the continuity equation provides an accurate estimation of mitral valve area in mitral stenosis before and after balloon valvotomy.


Arterial elastic properties in man: a comparison of echo-Doppler indices of aortic stiffness.

Non-invasive assessment of mechanical properties of the aorta may prove useful in the early detection of atheroma. We have evaluated several of the available echocardiographic indices using ability to detect age-related changes in putatively disease-free vessels as a measure of sensitivity to changes in aortic mechanical properties. Suprasternal imaging was used in 49 healthy non-smoking volunteers to measure minimum and maximum aortic arch diameters. Maximal flow velocities, with corresponding acceleration times and heart periods, were determined in the descending aorta in 24 of these subjects. Blood pressure was recorded non-invasively immediately after the echocardiographic study. Doppler derived measurements of aortic flow acceleration did not relate to age (P greater than 0.05). Three different 2D echo assessments of aortic distensibility, however, all showed a close relationship to age. Ep elastic modulus and Beta index (derived from different stress-strain mechanical relationships) were significantly related to age with r = 0.69 and 0.65 respectively. There were no significant effects of gender or left ventricular systolic function on these relationships. There was a tendency for the relationship between these distensibility indices and age more closely to fit an exponential than a linear relationship. We conclude that 2D echocardiographic assessment of aortic distensibility is able to detect sensitively changes in aortic mechanical properties. Even in the absence of risk factors for cardiovascular disease there is a marked reduction in aortic distensibility with increasing age.


The effects of thrombolytic therapy on the high-resolution electrocardiogram after myocardial infarction.

The effects of thrombolytic treatment was studied in 109 consecutive patients 9-11 days after their first acute myocardial infarction by high-resolution electrocardiography (ECG), 24 h Holter monitoring, exercise test and radionuclide ventriculography. Thirty-seven patients were treated with intravenous thrombolytic agents. Thrombolytic treatment was assessed by clinical criteria to be successful in 22 patients and probably successful in 12 patients. Thrombolysis failed in three patients and 72 patients did not receive thrombolytic treatment (control group). Measurements made on the high-resolution and filtered (60 Hz high-pass) vectormagnitude complex included the total duration, the duration of the potential less than 40 microV, the root mean square (RMS) voltage in 10 ms intervals over the first 50 ms and RMS voltage of the last 40, 50 and 60 ms. The filtered QRS duration was significantly shorter in reperfused patients compared with the control group (83 +/- 10 vs 89 +/- 12 ms; P = 0.017). In inferior infarcts (n = 57) the filtered QRS duration was 83 +/- 11 ms in reperfused and 89 +/- 10 ms in non-reperfused patients (P = 0.044), but in anterior infarcts (n = 52) there was no difference. The RMS voltage of the initial 50 ms of the QRS was higher in the reperfused than in non-reperfused anteroseptal infarcts (38 +/- 14 v 23 +/- 10 microV; P = 0.022). Patients successfully treated with thrombolytic agents within the first 2 h had higher RMS voltage of the terminal 40 ms of the QRS than patients treated within 2-4 h (38 +/- 17 vs 27 +/- 17 microV; P = 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)


Exercise habits and physical performance during comprehensive rehabilitation after coronary artery bypass surgery.

The effects of training as part of a comprehensive rehabilitation programme on exercise capacity and habits was studied in 171 male coronary artery bypass surgery patients randomized into a rehabilitation (R) (n = 93) and a reference, hospital-based treatment (H), group (n = 78). The rehabilitation programme started with a 2-day informative course before surgery and continued with a 3-week exercise-based course 2 months after surgery followed by a 2-day refresher 8 months post-operatively. The percentages of subjects having regular exercise were 22% and 10% pre-operatively, 42% and 38% 6 months and 46% and 38% 12 months after surgery in the R and H groups, respectively. The changes in the proportions observed in R and H groups were not significantly different. Total work during a bicycle exercise test increased from 38.9 +/- 24.3 kJ pre-operatively to 64.0 +/- 31.4 kJ 6 months (P less than 0.001) and to 70.0 +/- 35.7 kJ 12 months (P less than 0.001) post-operatively in group R and from 40.8 +/- 25.6 kJ to 57.3 +/- 26.6 kJ (P less than 0.001) and to 60.4 +/- 30.8 kJ (P less than 0.001) in group H, respectively. The increase from the pre-operative value was greater in group R than in group H both 6 (P = 0.03) and 12 months (P = 0.02) after surgery. Respective changes occurred in maximal work load, but the increase was significantly greater in group R than in group H only 12 months post-operatively.(ABSTRACT TRUNCATED AT 250 WORDS)


Improvement in left ventricular function after rapid weight loss in obesity.

The effect of rapid weight reduction on left ventricular function and blood pressure was studied in 34 obese subjects, who all weighed more than 30% above their ideal body weight. Subjects with co-existing hypertension (N = 15) and proven coronary artery disease (N = 9) were included to assess contributions from these factors. Blood pressure (BP) was measured both indirectly and by direct ambulatory intra-arterial methods. Radionuclide ventriculography was performed at rest and at exercise, before and after dieting, which was supervised in hospital (daily intake 330 kCal day-1 for 4 weeks). Dieting induced a weight loss of (mean +/- SD) 9.6 +/- 3.5 kg (P less than 0.0001) in the whole group. The mean daytime intra-arterial blood pressure fell from 157 +/- 23/90 +/- 15 to 144 +/- 21/85 +/- 13 mmHg (P less than 0.0001). The fall in ambulatory intra-arterial diastolic blood pressure did not reach significance in the normotensive and ischaemic groups. The mean resting left ventricular ejection fraction (LVEF) in the whole group showed a significant reduction after weight loss from 61 +/- 10% to 56 +/- 6% (P less than 0.005). Although the individual groups showed a fall in LVEF, the effect was most marked in the hypertensive group, from 65 +/- 9% to 57 +/- 5% (P less than 0.002). Before dieting none of the groups achieved a 'normal' 5% rise in LVEF above basal in response to exercise, the hypertensive and the ischaemic groups both showing non-significant falls. This phenomenon was, however, reversed after weight loss.(ABSTRACT TRUNCATED AT 250 WORDS)


Left ventricular filling pattern and pulmonary wedge pressure are closely related in patients with recent anterior myocardial infarction and left ventricular dysfunction.

To determine whether mitral flow velocity can be used to estimate mean pulmonary wedge pressure (PWP) in patients with left ventricular dysfunction, 50 patients with recent Q-wave anterior infarction and a reduced ejection fraction (less than 40%) underwent simultaneous pulsed-wave Doppler measurements of mitral flow and right heart catheterization. Doppler tracings and PWP were recorded at rest, after passive leg lifting (45 degrees) and (in 15 patients with increased PWP) after 5 mg sublingual ISDN. Significant correlations were found between the ratio of peak early to peak late diastolic velocity (E/A) and PWP (r = 0.83). Early diastolic deceleration and the ratio of the time velocity integral of atrial contribution to the total time velocity integral were also correlated to PWP (r = 0.80 and r = 0.79 (respectively). The E/A ratio was less than 1 in 25 patients and more than 1 in the remaining 25. An E/A ratio of at least 1 predicted a PWP of more than 20 mmHg with a sensitivity of 100% and a specificity of 86%. In all five patients, in whom the PWP was less than 20 mmHg at baseline and became greater with leg lifting, the E/A ratio changed from less than 1 to more than 1. After ISDN, changes in E/A ratio from more than 1 to less than 1 identified all 12 patients with a PWP falling below 20 mmHg. In conclusion, patients with recent Q-wave anterior infarction and a reduced ejection fraction mitral flow velocity-derived variables correlate with PWP representing a reliable index for the diagnosis of markedly increased PWP.(ABSTRACT TRUNCATED AT 250 WORDS)


Congenital complete heart block associated with QT prolongation.

The coexistence of congenital complete heart block and QT prolongation represents a special type of arrhythmia. The electrophysiological and clinical characteristics of this syndrome were studied in eight patients suffering from congenital AV block and QT prolongation. Data from 22 patients suffering from congenital complete heart block only, served as a control. In the study group, the appearance of a torsade de pointes type of ventricular tachycardia could regularly be observed and the tachycardial attack could usually be provoked by ventricular extrastimuli. The corrected QT time was markedly prolonged; on ventricular stimulation, at higher pacing rates the QT interval shortened, but remained significantly higher than in the control group. Syncopal attacks--with the character of polymorphic tachycardia--appeared in each patient of the study group while occurring in only three patients from the control group. Patients were given pacemaker implants (using a higher pacing rate) and long-term administration of beta-receptor blockers. The outcome was favourable; no ventricular tachycardia or syncopal attack was observed in the follow-up period.


Blood pressure changes during the game of squash.

The extent and pattern of the blood pressure response to the playing of squash was studied in five healthy volunteers using intra-arterial blood pressure recordings. Systolic pressure increased more than diastolic, but by only 18% of basal, peaking 5.2 +/- 2.3 min into the game (mean game duration 49 +/- 4 min). Thereafter there was a progressive decline, with reducing pulse pressure towards basal. There was a marked and significant increase in beat-by-beat blood pressure variability (P less than 0.01) and systolic peaks of up to 200 mmHg were recorded. A peak heart rate of 171 +/- 25 beats min-1 occurred at 20 min. These findings do not support the concept of a disproportionate and prolonged pressor response induced by playing squash. The possibility of high single-beat systolic peaks still justifies some caution in subjects at risk of arterial rupture.


Heart rate correction based on 'universal' regression equation--erroneous conclusions when studying cardiac mechanical function during stress.

This paper studies the possible errors associated with the use of a 'universal' regression equation in heart rate correction when used on mechanical cardiac function parameters. Systolic time interval rate correction, using Weissler regression equations, is taken as an example. The measurements were made during the isometric handgrip test, the orthostatic test, the Valsalva manoeuvre and the cold pressor test. It was observed that in most measurements rate correction caused a significant correlation between the changes in heart rate and the rate-corrected systolic time interval. It is considered that this new dependence between heart rate and the rate-corrected parameter makes the physiological interpretation of results more difficult. Therefore it is recommended that in stress provocations mechanical cardiac function parameters should not be rate-corrected by methods based on a 'universal' regression equation between heart rate and the mechanical function parameter.