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Biomedical subjects

C Johansson

Publications and source records attributed to C Johansson.

At least 235 records · Page 13Linked to original sources

Mouse monoclonal antibodies for experimental immunotherapy promotes killing of tumor cells.

Monoclonal antibodies (MAbs) produced against a human colon adenocarcinoma cell line, Colo-205, were tested in antibody-dependent macrophage-mediated cytotoxicity (ADMMC) assays. The antibodies C163, C215, C245 (IgG2a isotype); C151, C239, C241, C242 (IgG1); C152 (IgG2b); and C50 (IgM) were evaluated for their ability to promote killing of Colo-205 cells by thioglycollate-elicited peritoneal mouse macrophages. The concentrations of antibodies tested in ADMMC assays ranged from 1.0 ng/ml to 100 micrograms/ml, and the concentration at which 50% of tumor cells were lysed was used to characterize each antibody. Antibodies of the IgG2a isotype promoted the strongest macrophage-mediated tumor cell lysis effects in vitro. MAbs C215, C163, and C245 were also tested in nude mice which had been inoculated with Colo-205 cells. Tumor suppression was observed in mice injected with MAbs, supporting our ADMMC findings in vitro. Animals treated with MAbs had fewer and smaller tumors, and longer periods of latency between the inoculation of tumor cells and development of tumors, compared to mice sham-treated with PBS. However, in a study of established tumors, C215 antibody did not suppress tumor growth. Serum collected from MAb-treated mice promoted lysis of Colo-205 cells in ADMMC assays while serum from sham-treated mice did not.

Animals↗

Neonatal vasopressin antagonist treatment facilitates adult copulatory behavior in female rats and increases hypothalamic vasopressin content.

This study confirms that i.c.v. administration of Arg-vasopressin (AVP, 20 ng) inhibits copulatory behavior in female rats (lordosis response, LR) and demonstrates that this inhibitory effect is blocked by the vasopressin antagonist (3-mercapto-3-methylbutyryl-Tyr-[Me])arginine vasopressin (dPTyr(Me)AVP, 20 ng, i.c.v.). The effects of neonatal AVP antagonist treatment on adult female copulatory behavior, hypothalamic content of AVP and AVP-like immunoreactive (AVP-ir) neurons were examined. dPTyr(Me)AVP was given to female rats 1 microgram/animal/day s.c. from day 1 through to day 7 (day 0 = day of birth). The females were ovariectomized as adults and sexual receptivity activated by submaximal doses of estradiol plus progesterone. The LR was significantly facilitated in the neonatally dPTyr(Me)AVP treated females who also showed a higher content and an increased number of AVP-ir neurons in the suprachiasmatic nucleus compared to saline controls. Functional, biochemical and immunocytochemical evidence is provided that neonatal exposure to an AVP antagonist induces persistent changes in central vasopressinergic neuronal mechanisms.

Animals↗

The role of protein surface charges in ion binding.

Protein engineering is a means of probing the role of electrostatic interactions in protein functions; this elegant technique has been applied to the elucidation of electrostatic effects in enzyme catalysis. Here we show how the use of mutant proteins allows the determination of the contributions of individual charges to the free energy of ion binding to proteins. We have investigated the importance of three negatively charged side chains in the binding of Ca2+ to bovine calbindin D9K (ref.2): these are clustered around the calcium sites but are not directly involved as ligands. Each of these charges is found to contribute approximately 7 kJ mol-1 to the free energy of binding of two Ca2+ ions and to affect the cooperativity of Ca2+ binding. The influence of surface charges on ion binding to proteins may be more common than generally supposed and could have important consequences for protein function.

Calbindins↗

Fast horizontal electrophoresis. II. Development of fast automated staining procedures using PhastSystem.

The development of equipment for fast automated staining is described. It is possible to handle staining procedures with up to 20 steps and nine different solutions. To increase the reaction rate in the reaction chamber, the gels are rotated and high temperatures are used. The temperature in the reaction chamber is controlled between room temperature and 50 degrees C. Increased temperature, above 20 degrees C, generally results in faster staining and destaining. However, some reactions proceed better at a low temperature, including fixation of proteins with TCA, and the development step in silver staining, where increased temperatures cause a high background stain. Silver staining using acidic silver nitrate solution is preferred, due to easy preparation and good storage stability of the reagents. This method also causes little precipitation of silver on the walls of the reaction chamber. Silver staining is accomplished within one hour. Staining with PhastGel Blue is accomplished within 30 min.

Automation↗

Relationship between work and electromyographic activity during repeated leg muscle contractions in orienteers.

Contraction work (CW) was recorded for each of 200 repetitive isokinetic plantar flexions (1.05 rad.s-1) and knee extensions (1.57 rad.s-1) in 14 elite male orienteers. Simultaneous recordings of integrated electromyograms (iEMG) were obtained from the 3 parts of triceps surae and from 3 superficial portions of quadriceps femoris. CW in both muscle groups decreased significantly during the first 30 contractions (the fatigue phase), followed by a steady state level. The relative steady state level was higher for the plantar flexors (70 +/- 17%) than for the knee extensors (56 +/- 12%). For quadriceps a significant increase in iEMG occurred during the first 10 contractions followed by a decrease, whereas the iEMG of the plantar flexors showed a gradual decrease to the steady state level, which was similar for the two muscle groups (71-72%). The chosen expression of output/input balance (CW/iEMG) was constant throughout the plantarflexion test but decreased during the initial 20 knee extensions down to 82%. Thus, the fatigue phase of the knee extensions appeared to be divided into two; the first part had decreases in both CW and CW/iEMG and the second part with a decrease in CW alone. In contrast the plantar flexors only showed the characteristics of the second part.

Adult↗

Does a torn anterior cruciate ligament lead to change in the central nervous drive of the knee extensors?

Integrated surface electromyograms of the three superficial parts of the quadriceps and isokinetic knee extensor maximum torque and power production were recorded simultaneously and at different angular velocities in both legs in 11 male subjects with unilateral tear of the anterior cruciate ligament. The cross-sectional area (CSA) of the thigh and its muscular components were measured by computerized tomography. The principal findings were a small but significant decrease in quadriceps CSA on the affected side; a decreased active, but not passive, range of movement; decreased mechanical output, whether or not corrected for differences in CSA; and decreased electromyographic activity--particularly in rectus femoris. These findings suggest that the reason for the decreased maximum and total knee extensor performance seen in these patients is a change in knee joint receptor afferent inflow.

Adult↗

Effects of prostaglandins E2 and F2 alpha on motility of small intestine in man.

Interdigestive motility of the small intestine was examined in 23 fasted healthy volunteers following luminal administration of the prostaglandins E2 and F2 alpha. Motility was monitored by means of water-perfused catheters measuring intraluminal pressure changes. The registration points were located 25 cm apart, in the proximal duodenum, at the angle of Treitz, and in the jejunum. Prostaglandin E2 administered intraduodenally delayed the initiation of the subsequent activity front. The interval to the next activity front was prolonged by a dose of 1.0 mg prostaglandin E2 from 79.5 +/- 9.5 min to 137.1 +/- 5.0 min (P less than 0.01) and to 158.0 +/- 14.0 min by 2.0 mg prostaglandin E2 (P less than 0.05). Also, in four of seven experiments, a progressing activity front was arrested by 2.0 mg prostaglandin E2. Prostaglandin F2 alpha at 2.5 or 5.0 mg given intraduodenally induced bursts of contractions with a frequency of 17.7 +/- 0.8 contractions per minute and an amplitude of 10 to 110 mm Hg (P less than 0.01). In comparison, food intake produced irregular contractions at a frequency of 5.3 +/- 1.8 contractions per minute and an amplitude of 10 to 50 mm Hg (P less than 0.05). It is concluded that prostaglandin E2 delays the initiation of activity fronts in the duodenum. In contrast, prostaglandin F2 alpha changes the interdigestive motility pattern to one of intense contractile activity, which is different from the postprandial motility pattern.

Adult↗

Initial kinetic changes of prostaglandin E2-induced hyperplasia of the rat small intestinal epithelium occur in the villous compartments.

This study was performed to further identify the sequence of cell kinetics that occurs in the development of gastric and intestinal epithelial hyperplasia after orally administered prostaglandins of the E series. A high-dose, short-treatment schedule was used to examine the initial effects on kinetic parameters in the rat small intestinal epithelium. Groups of rats were killed after a single dose of oral prostaglandin E2 at 1 h after in vivo labeling with [methyl-3H]thymidine and during continued treatment at 6, 12, 24, 48, 72, and 96 h. As evidenced by autoradiography, the earliest change produced by prostaglandin E2 was an increased cellularity of the villous compartment (p less than 0.05 after 24 h). There was no change of labeling index of the villous compartment or of the leading edge of labeled cells within 24 h. At 48 h, the increased cellularity was accompanied by a significantly elevated labeling index of the villi. Throughout the study period no significant differences were observed between groups in the number of cells or labeling indices in the jejunal crypts, or in cellular input from the crypts to the villi. Epithelial turnover time in the placebo and treatment groups was 69 and 71 h, respectively. To exclude the possibility that prostaglandin E2 initially affects cell birth rate and mean cell cycle time, a metaphase blocker was given after 4 days of treatment in a second study. Animals were killed after 0, 0.5, 1.0, 1.5, and 2.5 h. The rate of entry into mitoses was 8.1% cells/h in controls compared with 8.2% cells/h in treated rats. The distribution of mitoses within crypts was identical in the two groups and the mean cell cycle time was 13.6 and 13.2 h, respectively. Also in this study there were trophic changes of the villi. It is concluded that the hyperplasia produced by oral prostaglandin E2 starts in the villi of the small intestine and is initiated by reduced cell exfoliation from the villous tips. Previously recorded retention of cellular elements in villi and crypts, increased cellularity of the proliferative compartments, and reduced mitotic index are secondary events.

Animals↗

Impaired pancreatico-biliary response to vagal stimulation and to cholecystokinin in human obesity.

We previously found that massively obese patients respond with less gastric acid secretion in response to vagal stimulation. This is compatible with the described association between experimental obesity and altered vagal function in the rat. To confirm this observation, the pancreatic and biliary responses to vagal stimulation were examined in nine nondiabetic obese patients against a background secretin infusion of 15 CU x h-1, and monitored after a subsequent injection of 75 IDU of cholecystokinin. Two separate marker perfusion systems were used in the stomach and duodenum, respectively, and blood samples were drawn for hormone analyses. In contrast to controls having normal body weight, the obese patients failed to respond with increments of pancreatic enzyme secretion and duodenal bile acids after stimulation with modified sham feeding. Cholecystokinin stimulated the pancreatic secretion of trypsin, amylase, and lipase, the emptying of bile acids, and the release of gastrin, but the patients' responses were only half that of the controls. In the resting state the obese had higher outputs of bile and pancreatic enzymes and higher plasma levels of pancreatic polypeptide compared with controls, but the pancreatic bicarbonate secretion rate was not different. The almost complete suppression of the basal gastric acid secretion by a low dose of intravenous (IV) secretin in controls did not occur in the obese. We conclude that massive obesity is associated with a reduced pancreatic and biliary response to vagal stimulation. Compared with controls, the digestive functions of the obese patients seem to be less sensitive to stimulation by exogenous cholecystokinin.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Mechanical output and iEMG of single and repeated isokinetic plantar flexions: a study of untrained and endurance-trained women.

Isokinetic plantar flexion peak torque (PT), contractional work (CW), and integrated electromyographic activity (iEMG) were analyzed in a group of untrained females and a group of female elite orienteers. Similar levels of PT, CW, and iEMG were observed in both groups during single maximum maneuvers (30-180 degrees s1). During repetitive (200) maximum plantar flexions, the untrained females showed a steeper and more pronounced decline in CW during the first 50 contractions than the orienteers. After this fatigue phase, all subjects could maintain a steady-state level of CW throughout the test. This level of CW was significantly (20%) higher in the orienteers than in the untrained females. The iEMGs of the gastrocnemii and soleus muscles decreased to significantly (10%) lower levels in the untrained women than in the orienteers. For both groups, the soleus muscle showed a significantly higher steady-state level of iEMG (10%) than mm. gastrocnemii. In the untrained group, CW/iEMG of m. triceps surae decreased to a plateau level significantly lower than in the orienteerers who could virtually maintain the initial CW/iEMG throughout the test. The differences observed could be explained by adaptations to performed endurance training in the orienteers.

Adult↗

Topical prostaglandin E2 in the treatment of chronic leg ulcers--a pilot study.

The results of treatment with topical PGE2 dispersed in hydrocolloid granules, in nine patients with chronic leg ulcers is reported. The healing process was evaluated by stereophotogrammetry, which enables objective measurements of ulcer area and volume. Ulcers in eight patients healed completely and that in the ninth, almost completely. These results suggest that topical PGE2 has a beneficial effect in the treatment of leg ulcers.

Administration, Topical↗

T-cell response to purified protein derivative after removal of Langerhans' cells from epidermal cell suspensions containing keratinocytes expressing class II transplantation antigens.

In a previous study we observed that human epidermal cell (EC) suspensions containing HLA-DR-expressing keratinocytes showed an amplified T-cell response to purified protein derivative (PPD). To evaluate further the possible immunological importance of class II transplantation antigens on keratinocytes we have compared the T-cell response to PPD in the presence of the following stimulator cells: EC suspensions from normal skin, or EC from tuberculin-reactive skin with or without removal of Langerhans' cells. The proliferation of purified T lymphocytes from peripheral blood in response to PPD in the presence of various concentrations of autologous EC was measured by [3H]thymidine incorporation on day 6. In 3 experiments out of 4 the EC from tuberculin-reactive skin, containing 28-76% HLA-DR-expressing cells as judged by immunocytochemistry (which also revealed fairly numerous HLA-DQ/-DP-expressing keratinocytes and a slight increase in CD36- and CD4- but not CD1-expressing cells), induced a more pronounced T-cell response to PPD than did normal EC. This was not the case in the fourth experiment, in which a small number of HLA-DR-(15%) and few if any HLA-DQ-/-DP-expressing keratinocytes were found. Immunomagnetic removal of CD1-reactive Langerhans' cells from the tuberculin-reactive EC suspensions resulted in a reduction of the T-cell response to PPD, in most cases down to background level (T cells alone + PPD). This study does not support the hypothesis that HLA-DR-expressing keratinocytes can in themselves act as antigen-presenting cells.

Adult↗

Peak torque and OBLA running capacity in male orienteers.

The isokinetic (30, 60, 120 and 180 degrees s-1) peak torque of m. quadriceps femoris and m. triceps surae, and the running velocity at the onset of blood lactate accumulation (VOBLA) were determined at three different occasions (November, April and September) during a year in 15 male élite orienteers. The amount and type of training for each individual was also registered. At the first test (November) there was no significant intra-individual correlation between peak torque of m. quadriceps and m. triceps surae. A negative correlation was found between VOBLA and peak torque of m. quadriceps at the lower velocities of angular motion, but not at the higher. The peak torque of m. triceps surae increased during the training period and decreased during the competition period back to the original level. Over the whole year the peak torque of m. quadriceps increased while no changes could be demonstrated within the two periods of the study. The running velocity at the onset of blood lactate accumulation increased during both training and competition periods. The changes in VOBLA were negatively associated with the changes in peak torque of m. quadriceps at 180 degrees s-1 during the training period. During the competition period the changes in VOBLA were, for the majority of the orienteers, negatively correlated to the ratio of quality:quantity running, i.e. to a high content of forest running.

Adult↗

Fatigue during dynamic muscle contractions in male sprinters and marathon runners: relationships between performance, electromyographic activity, muscle cross-sectional area and morphology.

Knee extensor fatigue and endurance during repeated maximum isokinetic manoeuvres were studied in five sprinters and five marathon runners, all males. Fatigue and endurance were expressed in terms of output (contractional work, CW) and input (integrated electromyograms, iEMG), and was related to muscle fibre composition of m. vastus lateralis and to cross-sectional area (CSA) of m. quadriceps. The initially performed contractional work was positively related to the calculated absolute type II fibre area of m. vastus lateralis, but not to the cross-sectional area of m. quadriceps. During the first 25 contractions, sprinters fatigued, with a slope of decline in contractional work that was correlated to the absolute type II fibre area, also after correction of performed work for CSA of m. quadriceps. As the electromyographic activity decreased less than the contractional work, a drastic fall in the CW/iEMG ratio was found in the sprinter group. Three out of five sprinters could perform less than 50 knee extensions, due to subjective discomfort in tested thighs. In contrast, all marathon runners could, after a minor similar decline in electromyographic activity and contractional work during the first part of the experiment, maintain a steady-state endurance level of CW and iEMG throughout the test (200 knee extensions). In conclusion, fatigue during isokinetic maximal knee extensions is closely related to the muscle structure of m. vastus lateralis.

Adult↗

Muscle pathomorphology in m. quadriceps of marathon runners. Early signs of strain disease or functional adaptation?

Muscle biopsies were obtained from m. vastus lateralis of five male marathon runners and five sprinters. The athletes performed their last competition at least 2 weeks prior to the investigation. No clinical signs of discomfort or injury were observed in the runners. The biopsy specimens from the sprinters showed an ordinary overall morphology, although fibre sizes varied considerably within and between the individuals. In contrast, in the marathon runners, structural deviations of pathologic value appeared in four out of five subjects. The fibre shapes were irregular, irrespective of fibre type, and internal architectural changes were common. An increased amount of connective tissue was found. A majority of fibres showed one or more central nuclei. A few fibres were flat and there were also evidence of fibre type grouping. The divergent picture might be an expression of repeated fibre injury with subsequent processes. If so, a peripheral, relative, arterial insufficiency is suggested as a possible aetiology. However, it cannot be ruled out that the deviations express a functional adaptation to the extreme endurance demands.

Adaptation, Physiological↗

Effects of prostaglandin E2 on disease activity, gastric secretion and intestinal permeability, and morphology in patients with rheumatoid arthritis.

The effects of oral natural prostaglandin E2 (PGE2) on symptoms, disease activity, and gastrointestinal functions in rheumatoid arthritis (RA) were studied in an open pilot trial. Twelve patients, six taking and six not taking non-steroidal anti-inflammatory drugs (NSAIDs), received 1 mg natural PGE2 three times a day for six weeks. The treatment was tolerated well and the only side effect noted was slightly looser stools in three patients. Half of the patients reported subjective improvement and none had aggravation of symptoms. The Ritchie articular index and several biochemical inflammation markers decreased and were significantly reduced at the end of the treatment period. The thickness of the small intestinal mucosa increased during the PGE2 treatment. The intestinal permeability pattern, measured by urinary excretion of polyethylene glycols (PEG 400), differed between the patients taking and not taking NSAIDs. The initially high urinary PEG 400 excretion values in the patients taking NSAIDs decreased and the initially low excretion values in patients not taking NSAIDs increased during the PGE2 treatment. The jejunal contents became sterile in 5/6 patients not taking NSAIDs and remained sterile in 1/6 patients taking NSAIDs at the end of the treatment. The treatment period was associated with a reduction of lactobacilli in patients not treated with NSAIDs. Thus the treatment appeared to decrease disease activity and to improve small intestinal functions in patients with RA, findings that need confirmation in a controlled trial.

Adult↗

Localization of antibody-secreting cells against human colon adenocarcinoma cell lines using the enzyme-linked immunospot assay.

The enzyme-linked immunospot assay was used to detect antibody-secreting cells from mice immunized against three human colon adenocarcinoma cell lines (Colo-205, SW 620, and SW 1116) on dishes coated with these cell lines. Antibody-secreting cells were detected by the development of a dark granular precipitate ('spot') using immunoenzymatic techniques where secreted antibody had bound to cell surface antigen. Spots can be detected by the naked eye and can be enumerated under low-power magnification. This method is specific, easily performed, versatile, and, together with conventional enzyme-linked immunosorbent assay techniques, allows for quantification of secreted antibody.

Adenocarcinoma↗