A triaxial dynamometer to monitor lateral bending and axial rotation moments during static trunk extension efforts.

OBJECTIVE: The purpose of the study was to describe a new static triaxial dynamometer designed to monitor the lateral bending and axial rotation moments during trunk extension efforts. BACKGROUND: Most studies on back muscle function using electromyographic spectral analysis have not controlled moments produced about the three orthopaedic axes during trunk extension efforts. Criteria to control lateral bending and axial rotation moments during extension efforts have not been proposed in the literature. METHODS: Fourteen healthy subjects performed three trunk extension ramp contractions (0-100% of the maximal voluntary contraction). Triaxial L5/S1 moments at 20%, 40%, 60% and 80% of the maximal voluntary contraction in extension were extracted. RESULTS: During the extension efforts, the lateral bending and axial rotation moments at L5/S1 increased significantly across the force levels and reached 6.2 Nm (SD: 6.6) and 6.1 Nm (SD: 4.5), respectively, at 80% of the maximal voluntary contraction. Tolerance limits were proposed to control these associated efforts in the context of the electromyographic analysis of back muscles. Relevance. Simultaneous measurement of lateral bending and axial rotation moments at L5/S1 during extension efforts might help to explain and control load sharing between back muscles during extension efforts.

Dynamometric assessment of the plantarflexors in hemiparetic subjects: relations between muscular, gait and clinical parameters.

The aims of this study were to investigate, in 16 subjects with hemiparesis, the plantarflexor muscle performance of the paretic side and to determine the level of the relationships between muscular parameters, clinical measures and gait performance. A Biodex dynamometric system was used to evaluate static and dynamic torques, power and maximal rate of tension development of the plantarflexor muscles. The clinical measures included the Fugl-Meyer assessment (FMA), the "Up & Go" test and an evaluation of ankle muscle tone. Velocity, cadence, stride length and gait cycle duration were determined for each subject at both comfortable and maximal safe speeds using foot contacts and videographic data. Results indicated that dynamometric values produced by the hemiparetic subjects were reduced in comparison to those reported for healthy subjects. Their torque-angle curves had a curvilinear shape which indicated pronounced decrease of torque for plantarflexion efforts at the beginning of the movement. Torques produced at different velocities of testing did not demonstrate significant differences (MANOVAs: p > 0.05) but power values were significantly different. Results also showed that all the selected muscular parameters (torque, power and maximal rate of tension development) were moderately to highly interrelated (0.65 < r < 0.94; p < 0.01) suggesting that a common factor of muscular performance was assessed. Furthermore, the dynamometric data were significantly associated with some of the clinical measures (sensation and lower limb motor control scores of the FMA) but were not related to the gait variables (Pearson's r < 0.45; p > 0.05). This last finding suggests that the relationship between plantarflexor strength and the level of gait performance in adults with stroke is complex. The relationship may be influenced by other factors such as muscular compensations within and between limbs and motor control impairments.

A motor reeducation program aimed to improve strength and coordination of the upper limb of a hemiparetic subject.

The objective of the study is to describe a new reeducation program based on a multi-directional and multi-articular dynamometer and to evaluate its applicability in one chronic right hemiparetic subject. The treatment sessions lasted 1 h and were conducted three times per week for a period of 8 weeks. During these sessions, the subject was asked to exert 10 repetitions of 16 torque combinations exerted at the shoulder, elbow and forearm or combined with handgrip exertion. The sequence of torques and force progressed from proximal to distal joints, and were realized in and out of the typical 'synergy patterns' described in this population. In addition, the levels of torque and force requested were increased progressively throughout the treatment period. The coordination of both upper extremities, tested using the finger to nose test, and the dexterity of the affected side, evaluated using the Box and Blocks assessment, tended to improve as treatments progressed. These results indicate the feasibility of this approach and suggest that it may be worthwhile examining the effectiveness of this approach on improving the functional performance of the upper extremity in a larger population of hemiparetic subjects.

Preloading and range of motion effect on plantarflexor muscle performance.

OBJECTIVE: To determine the effects of maximal preloading and range of motion (ROM) on the mechanical parameters of the plantarflexor muscles obtained while using the isotonic mode of testing of a Biodex dynamometer. DESIGN: A convenience sample of healthy subjects in the context of a descriptive comparative study. SETTING: Research laboratory in Canada. SUBJECTS: Fifteen volunteered subjects without history of injury or disorder to the right lower extremity. MEASUREMENTS: Four maximal isotonic tests were performed against a selected load of 27Nm. For the first two tests, the movement at the ankle ranged from -12 degrees (dorsiflexion) to +32 degrees (plantarflexion); one of these tests was preceded by a 2-second maximal preloading contraction, while the other was performed without preloading. For the other two tests, the ROM at the ankle was increased by 15 degrees of plantarflexion and thus ranged from -12 degrees to +47 degrees; again one of these tests was executed with preloading and the other without preloading. RESULTS: The four tests showed differences in the mechanical parameters (MANOVA p < .05). At angles of -10 degrees and +5 degrees, subjects produced higher torque and power but lower velocity values for the two tests preceded by a maximal preloading. The effect of ROM was demonstrated at +20 degrees where tests performed in a small amplitude reached a lower velocity than the corresponding tests performed in a larger amplitude. Based on the velocity profiles, the results also revealed that maximal preloading changed the selected isotonic movement of the Biodex dynamometer to an isoaccelerative movement characterized by high torque and power production. CONCLUSIONS: Isotonic assessment using the Biodex dynamometer provides different values of torque, velocity, and power depending on the testing conditions used. In clinical settings, it would be important to control these testing conditions.

Importance of Q fever in community acquired pneumonia.

Coxiella burnetii appears to be endemic in animals in the Mauricie region of Quebec, and causes some human cases of Q fever annually. Unlike in other rural areas, patients in this study experienced few respiratory symptoms. To determine whether C burnetii pneumonia is underdiagnosed, adults admitted to hospital for community acquired pneumonia were included in a one-year serological study. Significant immunofluorescent antibody (IFA) titres in four of 118 patients with pneumonia (fewer than 4%) were studied. Clinical presentation, standard laboratory tests and epidemiological data did not allow identification of these cases; however, Q fever increased during the warm months. There were no detectable complement fixing (CF) antibodies in these four cases. C burnetii causes few cases of pneumonia in Mauricie. IFA seems to be a more sensitive test than CF.

Sublethal levels of oxidant stress stimulate multiple serine/threonine kinases and suppress protein phosphatases in Jurkat T cells.

Sublethal concentrations of reactive oxygen intermediates including H2O2 can alter human T cell function and inhibit proliferative responses but relatively little is known about the effects of low levels of oxidant stress on signaling pathways. In the present study, we investigated whether the exposure of Jurkat T cells to micromolar concentrations of H2O2 might influence the activity of certain serine/threonine kinases and protein phosphatases important for T cell signaling as well as initiation of nuclear events. Jurkat cells treated with 100-200 microM H2O2 exhibited rapid increases in cytosolic protein kinase C (PKC) activity without detectable translocation of PKC to the membrane/particulate compartment. The stimulation of PKC activity by H2O2 was associated with an increase in the activation of kinases phosphorylating myelin basic protein (MBP), a substrate for mitogen-activated protein (MAP) kinase and RRLSSLRA (S6 peptide; a substrate for the approximately 90-kDa ribosomal S6 kinases). Optimal activation of MAP kinase in cells treated with H2O2 was preceded by increases in protein tyrosine phosphorylations and occurred at sublethal concentrations of H2O2 which did not markedly deplete intracellular ATP. Pretreatment of cells with the PKC inhibitors sangivamycin and H7 suppressed but did not block the stimulation of MAP kinase activity in response to H2O2 or phytohemagglutinin. The activities of both protein tyrosine phosphatase (PTP) and protein phosphatase 2A (PP2A) were reduced after H2O2 treatment of intact cells. Furthermore, kinetic studies showed that H2O2 was capable of suppressing the activities of PTP and PP2A before inducing optimal increases in MAP kinase activity. These results demonstrate that the exposure of T cells to sublethal levels of oxidant stress acutely stimulates the MAP kinase cascade and suggest that this activation may involve PKC-dependent and -independent pathways as well as inhibition of certain protein phosphatases.

Regulation of protein kinase enzymatic activity in Jurkat T cells during oxidative stress uncoupled from protein tyrosine kinases: role of oxidative changes in protein kinase activation requirements and generation of second messengers.

Prior studies have suggested that intracellular phosphorylation events and cellular redox mechanisms may interact in regulating a variety of cellular functions, including the transcriptional activation of gene expression. Increased activity of transcriptional factors NF kappa B and AP1 has been described in cells exposed to oxidative stress and following the direct stimulation of protein kinase C (PKC) by phorbol diesters. However, the mechanisms that may contribute to redox regulation of PKC are unknown. We studied the expression of PKC activity and several second messengers in human Jurkat T cells exposed to oxidative stress in the form of H2O2. Micromolar concentrations of H2O2 rapidly induced increased cytosolic PKC enzymatic activity in Jurkat T cells that was associated with a marked arrest of cellular proliferation. The increase in cytosolic PKC activity in cells treated with H2O2 was accompanied by elevations in intracellular free calcium ([Ca2+]i), generation of inositol phosphates, and release of arachidonic acid. Functional studies showed that H2O2 enhancement of cytosolic PKC activity required phospholipase C activity but was not primarily mediated by arachidonic acid. The response of PKC to oxidative stress displayed a lack of Ca2+ dependence and was uncoupled from the activity of protein tyrosine kinases (PTK). Furthermore, the reduced activation requirements of PKC from cells treated with H2O2 were associated with shifts in elution profiles of PKC enzymatic activity after Mono-Q chromatography. These shifts appeared to represent intrinsic changes in the conformation of PKC induced by oxidative stress because western blotting failed to reveal any PKC cleavage products or reductions in native PKC alpha or beta. These findings indicate that oxidative regulation of intracellular events can intersect phosphorylation events mediated by PKC through the release of second messengers as well as direct changes in PKC activation requirements. Moreover, redox regulation of PKC is distinct from T cell receptor signaling in that the activity of PKC is uncoupled from the regulatory influences of PTK.

Q fever in Quebec (1989-93): Report of 14 cases.

Q fever, a zoonosis acquired by inhalation of the rickettsia Coxiella burnetii, is rarely diagnosed in Canada. The world incidence has been increasing since 1960, because of progressive dissemination of this microorganism in animal populations, particularly domestic ruminants. Some recent outbreaks were caused by cats. Of 14 cases reported in Quebec between 1989 and the beginning of 1993, nine occurred successively in an 18-month period in the rural region surrounding Trois-Rivières, after contact with livestock or cats. These cases are reported here, with the results of serological screening of the workers of an abattoir where one of the cases worked. Five additional cases reported in Quebec during the same period are briefly reviewed.

A static dynamometer measuring multidirectional torques exerted simultaneously at the hip and knee.

The function of a static dynamometer measuring torques exerted simultaneously in the different anatomical planes of the hip (flexion-extension, abduction-adduction and internal-external rotation) and knee (flexion-extension) is described. Muscular torques were calculated in real time using a desktop computer from measurements of orthogonal forces applied at two locations and the lever arm values measured in each subject. The reliability of the force transducers was explored by examining their output, using calibrated weights, on three different days. The results were identical over this period of time, indicating that the transducers are highly reliable. A mechanical simulator of a lower limb was constructed to generate specific or combined torques of known values at the hip and knee. The torques measured by the dynamometer were found to be highly concordant with the known torques applied by the simulator, indicating that the torque measurements were valid. The usefulness of the dynamometer is demonstrated by evaluating the activity of the rectus femoris and biceps femoris muscles during static efforts exerted in various directions at the hip. In addition, the mechanical action of biarticular muscles at the hip was evaluated by quantification of hip torques during efforts exerted at the knee. From these results, it has been concluded that the present biarticular and multidirectional dynamometer is a valid, reliable and precise instrument that may prove to be useful in evaluating the muscular function of the lower limb.

Progression of colorectal cancer is associated with multiple tumor suppressor gene defects but inhibition of tumorigenicity is accomplished by correction of any single defect via chromosome transfer.

Carcinogenesis is a multistage process that has been characterized both by the activation of cellular oncogenes and by the loss of function of tumor suppressor genes. Colorectal cancer has been associated with the activation of ras oncogenes and with the deletion of multiple chromosomal regions including chromosomes 5q, 17p, and 18q. Such chromosome loss is often suggestive of the deletion or loss of function of tumor suppressor genes. The candidate tumor suppressor genes from these regions are, respectively, MCC and/or APC, p53, and DCC. In order to further our understanding of the molecular and genetic mechanisms involved in tumor progression and, thereby, of normal cell growth, it is important to determine whether defects in one or more of these loci contribute functionally in the progression to malignancy in colorectal cancer and whether correction of any of these defects restores normal growth control in vitro and in vivo. To address this question, we have utilized the technique of microcell-mediated chromosome transfer to introduce normal human chromosomes 5, 17, and 18 individually into recipient colorectal cancer cells. Additionally, chromosome 15 was introduced into SW480 cells as an irrelevant control chromosome. While the introduction of chromosome 17 into the tumorigenic colorectal cell line SW480 yielded no viable clones, cell lines were established after the introduction of chromosomes 15, 5, and 18. Hybrids containing chromosome 18 are morphologically similar to the parental line, whereas those containing chromosome 5 are morphologically distinct from the parental cell line, being small, polygonal, and tightly packed. SW480-chromosome 5 hybrids are strongly suppressed for tumorigenicity, while SW480-chromosome 18 hybrids produce slowly growing tumors in some of the animals injected. Hybrids containing the introduced chromosome 18 but was significantly reduced in several of the tumor reconstitute cell lines. Introduction of chromosome 5 had little to no effect on responsiveness, whereas transfer ot chromosome 18 restored responsiveness to some degree. Our findings indicate that while multiple defects in tumor suppressor genes seem to be required for progression to the malignant state in colorectal cancer, correction of only a single defect can have significant effects in vivo and/or in vitro.

Expression of hepatocyte and oval cell antigens in hepatocellular carcinomas produced by oncogene-transfected liver epithelial cells.

We have established an in vivo/in vitro system in which epithelial cells ("oval cells") isolated from livers of rats fed a carcinogenic diet for a very brief period are placed in culture and transfected with an oncogene. Injection s.c. into nude mice of oval cells transfected with the activated c-Ha-ras (EJ oncogene) produces tumors with morphological features of differentiated hepatocellular carcinomas. Using monoclonal antibodies that can recognize hepatocyte, oval cell, and tumor antigens, we investigated the expression of these antigens in oval cells in culture, transfected with either the EJ oncogene or the normal c-Ha-ras allele and in tumors derived from the oncogene-transfected cells. We show that EJ-transfected cells and most particularly the tumors they produce expressed hepatocyte and oval cell antigens not detectable in untransfected cells or cells transfected with the normal c-Ha-ras gene. Furthermore, we found that in cloned tumor cells, the expression of hepatocyte antigens could be induced by changes in culture conditions and was accompanied by a decrease in the expression of oval cell markers. Trabecular hepatocellular carcinomas had higher reactivity toward monoclonal antibodies recognizing hepatocyte antigens while tumors with glandular architecture reacted predominantly with monoclonal antibodies against oval cells. We conclude that, in addition to its tumorigenic effect, the EJ oncogene induced the differentiation of tumor cells toward the hepatocyte lineage. In addition, the data provide further confirmation that oval cells can serve as progenitors of differentiated hepatocellular carcinomas.

[Prognostic value of HIV serum antigens]. / Valeur pronostique de l'antigénémie HIV.

Human immunodeficiency virus (HIV) antigens can be detected during early infection but usually disappear afterwards. Their later reappearance in serum seems to herald a greater virus replication and a poor prognosis. The prognostic value of antigen detection at the first clinical visit was assessed among 37 symptomatic patients. Patients with detectable antigenemia at the first visit progressed more rapidly towards the acquired immunodeficiency syndrome (p less than 0.0001). Similarly, patients with detectable antigens at the time of a Pneumocystis carinii pneumonia had a shorter survival than patients without detectable antigens (p less than 0.0001). Four months after the hospital admission date, the survival rate was 31% among patients with detectable antigens and 100% among patients without detectable antigens (p less than 0.0001). In two of four patients tested, serial HIV antigens determinations suggested a spontaneous decrease of antigen titers during the terminal phase of the disease.

Transforming activity of DNA from rat liver tumors induced by the carcinogen methyl(acetoxymethyl)nitrosamine.

Altered c-Ha-ras genes have been frequently detected in the DNA of spontaneous or chemically induced mouse liver tumors. To determine if ras gene mutation is a frequent event during liver carcinogenesis in rats, we examined the transforming activity of DNA from liver tumors that developed in rats injected with methyl(acetoxymethyl)nitrosamine (DMN-OAc) after a partial hepatectomy. Three weeks after the injection of DMN-OAc, rats were fed a diet containing phenobarbital. This carcinogen acts only on replicating liver cells. Six of eight tumor DNAs induced the transformation of NIH 3T3 cells. The transforming activity was stable upon a second round of transfection, and the transformants were tumorigenic in nude mice. Southern blot analysis of transformant DNAs showed that the transforming activity was not due to the acquisition of a ras (Ha, Ki, or N), neu, myc, A-raf, v-raf, erbA, or erbB gene of rat origin. Several transformants' restriction enzyme sensitivity was analyzed, and their activity indicated that similar transforming sequences were present in at least two tumors and that one tumor contained two different transforming sequences. These results suggest that during hepatocarcinogenesis induced in rats by DMN-OAc, alterations in the ras gene family occur infrequently or not at all and that several different genes (which are not homologous to common oncogenes) become activated and are capable of transforming NIH 3T3 cells.

Growth in culture and tumorigenicity after transfection with the ras oncogene of liver epithelial cells from carcinogen-treated rats.

Two epithelial cell lines designated LE/2 and LE/6 were established from cells isolated by centrifugal elutriation from the livers of carcinogen-treated rats. Both cell lines exhibit some characteristics of fetal liver cells, such as the expression of the 2.3-kilobase alpha-fetoprotein mRNA, aldolase A, and lactate dehydrogenases 4 and 5. Primary cultures contain gamma-glutamyl transferase-positive cells which do not proliferate in vitro. After the first passage, the LE/2 and LE/6 cell lines are uniformly gamma-glutamyl transferase negative. Neither cell line is transformed as assayed by morphology, anchorage-independent growth, or tumor formation in nude mice. By the 50th passage, LE/6 cells form numerous colonies in soft agar in the presence of epidermal growth factor, while no colonies grow in medium lacking this growth factor. Clonal cell populations derived from five epidermal growth factor-induced soft agar colonies were not tumorigenic in nude mice. This indicates that, although epidermal growth factor-responsive late passage cells had acquired some of the phenotypic properties commonly associated with tumor cells, these cells were not fully transformed. Transformation of LE/6 cells was accomplished by transfection of the rasH oncogene (EJ). Subcutaneous inoculation of rasH (EJ)-transfected LE/6 cells produced tumors at the site of injection with histological features of moderate to well-differentiated trabecular hepatocellular carcinomas. Tumor cell lines derived from the nude mouse tumors are gamma-glutamyl transferase positive and express alpha-fetoprotein mRNA. One clonal cell line expresses both alpha-fetoprotein and albumin mRNA. These results show that nonparenchymal liver epithelial cells transfected with an activated oncogene can give rise to differentiated hepatocellular tumors similar to those induced in livers of rats fed a carcinogenic diet.

Sequential protooncogene expression during rat liver regeneration.

When growth is stimulated in the normally quiescent adult rat liver by partial hepatectomy, steady state levels of messenger RNAs (mRNAs) for c-fos, c-myc, and p53 increase sequentially during the prereplicative phase which precedes DNA synthesis. Levels of c-fos mRNA are elevated at least 4-fold within 15 min after partial hepatectomy and decrease rapidly by 2 h; c-myc mRNA reaches maximal levels (5-fold over normal) between 30 min and 2 h after the operation. A second, transient phase of expression for both c-fos and c-myc occurs around 8 h after partial hepatectomy. p53 mRNA levels increase between 8 and 12 h after the operation (5-fold over normal) and are reflected in an elevation of steady state levels of p53 protein between 12 and 15 h after partial hepatectomy. The levels of ras p21 protein increase much later at a time of active DNA replication and cell division. Actinomycin D injected at the time of partial hepatectomy blocks the increase in c-myc at 2 h but has no effect on c-fos mRNA levels. Actinomycin D injected at 6 h only partially blocks the increase in c-myc and p53 mRNA at 8 h but does not affect c-fos mRNA. Our results suggest that the transient and sequential expression of protooncogenes during the prereplicative stage of liver regeneration is likely to reflect events associated with entry and progression of hepatocytes into the cell cycle and can serve as markers for identifying specific humoral factors involved in liver regeneration.

Proto-oncogene expression and growth factors during liver regeneration.

When growth is stimulated in normally quiescent hepatocytes, steady-state levels of c-fos, c-myc, and p53 mRNAs increase sequentially and transiently before DNA replication. C-fos mRNA increases almost immediately after partial hepatectomy and decreases by 2 hr; c-myc mRNA reaches maximal levels between 30 min and 2 hr. In contrast, the p53 mRNA increase corresponds to the G1/S transition, and mRNAs from c-ras genes are elevated later, coinciding with DNA replication and mitosis. p53 and p21 proteins are elevated when their mRNAs are more abundant. This regulated response suggests that these genes either control key steps in the cell cycle or are responding to humoral or internal growth factors acting at specified growth stages. We propose that hepatocytes go through a "priming" stage during the first four hours after partial hepatectomy and that their progression through late G1, is likely to be controlled by autocrine or paracrine mechanisms, which may account for the precisely regulated growth of the liver after partial hepatectomy. Transforming growth factor beta (TGF beta) is a potent inhibitor of DNA synthesis in normal hepatocytes in vitro. We show that TGF beta mRNA increases in the regenerating liver at the time of hepatocyte DNA synthesis and mitosis. In normal or regenerating liver, the mRNA for this growth factor is contained in nonparenchymal cells but not in hepatocytes. We suggest that TGF beta may be a component of a paracrine regulatory loop that controls hepatocyte replication.

Expression of c-Ki-ras, c-Ha-ras, and c-myc in specific cell types during hepatocarcinogenesis.

We examined the expression of six proto-oncogenes in (i) whole rat liver and isolated liver cell populations during the course of hepatocarcinogenesis induced by a choline-deficient diet containing 0.1% ethionine and (ii) fetal rat liver at different stages of development. The abundance of c-Ki-ras, c-Ha-ras, and c-myc transcripts in polysomal polyadenylated RNA from liver cells increased by 2 weeks after the start of the carcinogenic diet. c-Ki-ras and c-myc expression remained elevated during the 35 weeks of the diet, whereas c-Ha-ras transcripts increased transiently. A primary tumor sampled at 35 weeks after the carcinogenic diet was started contained high levels of both c-Ki-ras and c-myc RNA. The abundance of c-src transcripts was unchanged throughout carcinogenesis; c-abl and c-mos transcripts were not detected in either preneoplastic or neoplastic livers. To determine which cell types within the liver contained proto-oncogene transcripts, we isolated hepatocytes, oval cells, and bile duct cells from normal and preneoplastic livers. The results indicate that proto-oncogenes are expressed differentially in these cell types during hepatocarcinogenesis and that the expression of c-Ki-ras and c-myc is high in oval cells throughout carcinogenesis. In developing livers, c-Ki-ras, c-Ha-ras, and c-myc transcript levels were high at 17 days of gestation but reached the low values characteristic of adult rat livers between 20 days of gestation and 3 days after birth.

Regulated transcription of c-Ki-ras and c-myc during compensatory growth of rat liver.

We examined the transcription of six cellular oncogenes during the process of compensatory growth in rat liver after partial hepatectomy. We have previously reported that transcripts of c-rasH are elevated during regenerative growth of the liver. We now report that transcripts of c-rasK and c-myc genes are significantly elevated after partial hepatectomy, whereas transcripts of c-abl and c-src are essentially unchanged and transcripts of c-mos are undetectable in either normal or regenerating rat liver. In liver regeneration after partial hepatectomy or chemical injury, changes in c-myc transcripts occur before DNA synthesis. The elevation of c-myc and c-ras transcripts is sequential in that highest levels of c-myc transcripts were detected 12 to 18 h after partial hepatectomy, whereas the levels of c-rasH and c-rasK were maximal by 36 to 48 h. Transcripts of all three activated oncogenes returned to their basal levels by 96 h.