Josef G. Grohs

Antibodies to heat shock protein 90 in osteosarcoma patients correlate with response to neoadjuvant chemotherapy

Autoantibodies to the heat shock protein 90 (Hsp 90) have been reported as prognostic marker in breast cancer patients. Sera from 20 high-grade osteosarcoma patients were tested at the time of diagnosis by enzyme-linked immunosorbent assay. Presence of anti-Hsp90 antibodies correlated with a better response to neoadjuvant chemotherapy (P< 0.01), whereas the absence correlated with development of metastases. These data suggest that anti-Hsp90 antibodies might be of predictive value in human osteosarcoma.

Serum Antibodies against the Heat Shock Protein 60 are Elevated in Patients with Osteosarcoma

Osteosarcoma is the most frequent malignant bone tumor, mainly occurring in the second and third decade of life. Diagnosis is limited to clinical symptoms, radiology and histology, but so far no diagnostic laboratory tests are available. Heat shock proteins (hsp), highly conserved proteins performing vital intracellular chaperoning functions and preventing cells from death, have been shown to be involved in tumor immunity. We analyzed 75 sera from 23 patients with high-grade osteosarcoma, 8 patients with chondrosarcoma, 10 patients with Ewings sarcoma, 5 patients with soft tissue sarcoma, 11 patients with benign bone tumors at the time of diagnosis and from 18 healthy controls with an indirect one-site enzyme linked immunosorbent assay (ELISA) for the presence of anti-hsp60 and 70 antibodies. In these assays 10/23 osteosarcoma patients (43%) had anti-hsp60 antibodies with a mean +/- S.D. titer of 0.382 +/- 0.243 U/ml. Only one of the 18 healthy controls (1/18, 5.6%; titer 0.22 U/ml), two of the Ewings sarcoma patients (2/10, 20%; titer 0.2 +/- 0.09 U/ml), two of the patients with a benign bone tumor (2/11, 18%; titer 0.22 +/- 0.16 U/ml) and one of the chondrosarcoma patients (1/8, 12.5%; titer 0.14 U/ml) were positive, whereas all others, including all soft tissue sarcomas were negative throughout. Anti-hsp60 antibodies in patients with osteosarcoma are therefore significantly increased (p < 0.05). 19/23 (83%) of osteosarcoma biopsy specimens expressed hsp60 immunohistochemically and all specimens from patients with a positive anti-hsp60 serum titer expressed hsp60. The level of the anti-hsp60 antibodies did not correlate with clinical parameters such as response to preoperative chemotherapy, duration of symptoms, age, gender, tumor size, serum alkaline-phosphatase levels and metastases. Although no difference in anti-hsp70 antibodies could be observed between sera from patients and healthy controls, a positive correlation was found for the presence of anti-hsp70 serum antibodies and lung metastases at the time of diagnosis in osteosarcoma patients. These data suggest an increase of anti-hsp60 antibodies at the time of first diagnosis of osteosarcoma. These findings should therefore give rise to further investigations on a group of new markers for the diagnosis of osteosarcoma.

Cardiac and hemodynamic effects of the selective bradycardic agent KC 8857 during exercise-induced myocardial ischemia.

The effects of an in vitro bradycardic agent without negative inotropism, KC 8857 (3,7-di-(cyclopropylmethyl)-9,9-tetramethylene-3,7-diazabicyclo-[3.3.1]- nonane dihydrochloride), were tested in chronically instrumented dogs in a model of exercise-induced myocardial ischemia. KC 8857 was i.v. infused during critical stenosis of the circumflex branch of the left coronary artery which led to exercise-induced myocardial dysfunction. KC 8857 caused a decrease in heart rate, left ventricular dp/dtmax and calculated myocardial oxygen demand at rest and during exercise. Since positive dp/dtmax values at a given heart rate were not altered by KC 8857 it may be assumed that myocardial function was restored mainly by the decrease in heart rate.

Detection of orthopaedic prostheses at airport security checks

We studied the detection of joint replacements at airport security checks in relation to their weight, using two types of detector arch. A single-source, unilateral detector showed different sensitivities for implants on different sides of a test subject. All implants weighing more than 145 g were detected by one of the arches. The degree of detection was directly related to the logarithm of the weight of the prosthesis in patients, with a linear correlation (r2 = 0.61). A bilateral arch detected all prostheses weighing over 195 g. With their usual sensitivity settings many joint replacements were detectable; an identification pass containing the site and weight of such prostheses would help to avoid the need for body-search procedures.

Simultaneous assessment of cardiac output with pulsed Doppler and electromagnetic flowmeters during cardiac stimulation

The aim of this study was to test the accuracy of cardiac output assessment by Doppler and electromagnetic flowmetry in dogs during states of (1) marked enhancement in cardiac output, which was obtained by means of either isoprenaline infusion or treadmill exercise, or (2) reduction in cardiac output obtained by administration of phenylephrine. Additionally, in vitro comparisons were undertaken between Doppler and electromagnetic flow-probes and assessment of flow by direct volumetric measurement. These in vitro experiments showed a good correlation between timed volume collections and electromagnetic flow assessment up to high flow velocities. Doppler flow measurements underestimated the flow at high velocities. In both the resting dog and after phenylephrine, that is, at states with low heart rate and cardiac output, the waveforms of electromagnetic flow and Doppler velocity were similar for both phasic and mean flow, respectively. During states of cardiac stimulation Doppler flow showed a decrease in maximum velocity in the ascending aorta. Due to this decrease in peak flow velocity, mean Doppler blood flow did not increase despite of increased heart rate. This result cannot be explained on the basis of the deviation of Doppler measurements at high velocities in the in vitro experiments. Although our results are in contradiction with earlier studies, electromagnetic assessment seems to be more reliable in blood flow measurements in the ascending aorta. Hence, Doppler flow measurements should not be used uncritically for such quantitative flow assessment in large vessels as determination of cardiac output.

Oral NG-nitro-L-arginine in conscious dogs: 24 hour hypertensive response in relation to plasma levels

SummaryHaemodynamic changes after oral administration of 30 mg/kg NG-nitro-L-arginine (L-NNA) were studied in conscious chronically instrumented mongrel dogs throughout a 24 h observation period in order to evaluate the long-term efficacy of L-NNA-induced inhibition of endothelium-dependent relaxation and its relation to plasma L-NNA level. Diastolic blood pressure remained elevated for the entire 24 h observation period, but systolic blood pressure was raised only up to the 6 h value. The hypertensive response was accompanied by bradycardia. The increase in blood pressure and the plasma L-NNA level both reached their maxima at 3 h. The plasma L-NNA level at the end of the observation period was diminished by only 21.7% with respect to the maximum increase, whilst the maximum increase in mean arterial blood pressure was attenuated by 72.2% at 24 h. These data show a dissociation between plasma L-NNA level and the respective blood pressure.

Cardiovascular effects of flesinoxan in anaesthetized and conscious dogs

SummaryStimulation of 5-HT1A receptors is known to decrease the arterial blood pressure in anaesthetized rats, cats and dogs. We investigated the hypotensive activity of flesinoxan (0.1 + 0.2 + 0.7 μmol/kg), a 5-HT1A-receptor agonist, in dogs anaesthetized with either morphine and pentobarbital or enflurane and also in the conscious state. Flesinoxan led to a decrease in arterial blood pressure in anaesthetized, but not in conscious dogs. In the conscious state the marked increase in heart rate, which can be taken as an indicator of sympathetic tone, may have masked the consequences of vasodilatation. These different haemodynamic responses to flesinoxan may be dependent on side effects of the drug in the conscious dogs, in particular hyperventilation and salivation combined with anxiety, and on the magnitude of the decrease in baroreceptor reflex activity during anaesthesia with morphine and pentobarbital on the one hand and enflurane on the other hand.

Antiischemic effects of tedisamil in conscious dogs

SummaryThe antiischemic effects of tedisamil were investigated in chronically instrumented conscious dogs. Heart rate, arterial blood pressure, left ventricular dP/dtmax, left ventricular end-diastolic pressure, and regional myocardial shortening in the ischemic and normal myocardium, assessed by sonomicrometry, were measured. Reversible regional myocardial dysfunction was achieved by graded treadmill exercise in the presence of a critical coronary stenosis, which was produced by partial inflation of a hydraulic occluder. Regional myocardial dysfunction, which was observed befere the intravenous drug administration, was almost completely abolished by tedisamil (180 μg/kg/5min). The antiischemic effect of tedisamil is due to a reduction in heart rate, which leads to a decrease in left ventricular dP/dtmax and myocardial oxygen demand in the presence of a prolongation of diastole and thus coronary perfusion time. Clinical evaluation will test the value of this drug for the treatment of stress-induced symptomatic and asymptomatic angina pectoris in humans.

Effects of flestolol, an ultra-short acting-β-adrenoceptor antagonist, on hemodynamic changes produced by treadmill exercise or isoprenaline stimulation in conscious dogs

The β-blocking activity of flestolol was established during increasing isoprenaline infusions and during graded physical exercise in conscious, chronically instrumented dogs. After a control cycle, flestolol was infused at three doses (1, 2.67, and 10 μg/kg/min). Flestolol has an extremely short half-life, demonstrated by an 83% loss of effect within 25 min. Thus, flestolol allows easy titration of the effect, which might be a valuable property for its use in the treatment of critically ill patients. Due to the difference between pure β-adrenergic stimulation and the much more complex regulation of circulation during exercise, the hemodynamic response to flestolol elicited marked differences between both set ups. Flestolol shifted the dose-response curves of isoprenaline-induced changes in heart rate, positive left ventricular dp/dtmax and diastolic arterial pressure dose dependently to the right, while its main effect during exercise was a decrease in positive left ventricular dp/dtmax. Thus, testing of β-adrenoceptor blockers using isoprenaline-induced tachycardia leads to an overestimation of potency and therefore is not appropriate to predict the clinical efficacy of these drugs to prevent stress- or exercise-induced in-creases in heart rate and hence myocardial oxygen-demand.

Contractile function of human veins after long-term storage in different media.

We wished to determine the preservation of contractile function of human saphenous veins during long-term storage (36 and 96 h) at low temperatures in different solutions. Two crystalloid solutions (Krebs-Henseleit and Bretschneiders cardioplegic solution), as well as heparinized blood and albumin solution as used in cardiac surgery were compared. Contractile function of human saphenous vein ring segments was tested in a vessel myograph. Potassium chloride (KCl 80 mM) was used to achieve receptor-independent maximal contraction; contractility to norepinephrine (NE) was tested in a concentration-dependent manner (10(-8)-10(-6)M). Nitroglycerin (NTG) was used to test vascular relaxation. Thirty-six hours of storage in 5% human albumin abolished responses to KCl and to NE. At this time, contraction of vein ring segments stored in heparinized blood was decreased to 24% in response to KCl and to 16% in response to NE. After 36-h storage in Bretschneiders cardioplegic solution, contractile function was not significantly decreased. The contraction was still 82% after KCl and was unchanged after NE. After 96 h, the contractile response was markedly impaired in all vein ring segments, and no relevant differences were evident between the solutions. In all experiments, NTG caused complete relaxation. Because Bretschneiders cardioplegic solution preserved vascular contractile function for 36 h, the implementation of this solution might enable long-term storage of human vessels for transplantation, bypass grafts, and in vitro experiments.