To support clinical pharmacokinetic studies in cancer patients, sensitive and specific methods for measuring 4-[1-(4-cyanobenzyl)-5-imidazolylmethyl]-1-(3-chlorophenyl) piperazinone (I), a farnesyl transferase inhibitor (FTI), in human plasma and urine were developed and validated. The methods are based on high-performance liquid chromatography (HPLC) with atmospheric pressure chemical ionization (APCI) and tandem mass spectrometric (MS/MS) detection in the positive ion mode using a heated nebulizer interface. Drug and internal standard were isolated from plasma or basified urine using automated solid-phase extraction on cyano cartridges. The organic extracts were dried, reconstituted in aqueous acetonitrile and injected into the system. Chromatographic separation of I and internal standard (IS) was achieved using a BDS Hypersil C8 analytical column, with a mobile phase consisting of acetonitrile:methanol:water (50:4:46) and trifluoroacetic acid (0.05%) at a flow rate of 0.6 ml/min. MS/MS detection was performed on a PE-Sciex API 300 tandem mass spectrometer operated in selected reaction monitoring mode. The parent->>product ions monitored were m/z 406-->195 for analyte I and m/z 448-->195 for the internal standard. Unusual in this method is that quantitation is accomplished using a secondary product ion, m/z 195, of drug I and IS. The assays were validated over the concentration range of 0.5-1000 ng/ml (1.2 nM to 2.5 microM, respectively) in plasma, and 2.5-500 ng/ml (6.2 nM to 1.23 microM) in urine. Accuracy was within +/-10% of nominal concentration at all levels in urine, and all but the lowest standard in plasma (+/-14% at 0.5 ng/ml). Intraday precision (expressed as coefficients of variation, CVs) for standard replicates and interday precision for quality control (QC) samples were less than 8% at all concentrations in both matrices. Detailed descriptions of the extraction procedure and analytical methodology used in the assay of I in plasma and urine are presented. This procedure may have utility in the quantitation of other imidazole-based FTIs with cyanobenzyl substructures.

Blood plasma free radical lipid oxidation was studied in 98 patients with acute dysentery (AD) and 64 patients with food toxinfection (FTI). Free radical lipid oxidation was assessed by the intensity of chemiluminescence and the level of lipid peroxides. Activation of blood plasma lipid peroxidation was established with relation to a clinical variant of the disease. The effect of indomethacin on the parameters of free radical lipid oxidation was noted in patients with AD and FTI.

A computer model of radiometric scene simulation for simulated spectra for pollution clouds in complicated environment is proposed. The model is used to introduce the effects of an actual hazardous pollution clouds, such as (CH3)2CHO(CH3)FPO or (CLCH2CH2)2S, into exiting measured background spectra by passive Fourier transform infrared spectrometer (FTIS). The simulated results agree well with the experimental results.

Rheopolycardiological investigation of 76 patients with food toxinfections (FTI) showed that phasic changes of systolic indices in these diseases developed as a result of extracardiac hemodynamic disorders and were of adaptive nature. Strophanthin administration to 20 similar patients at the height of disease brought about no statistically significant changes in systolic indices. It was concluded that the use of cardiac glycosides was inappropriate for the treatment of FTI.

Infrared spectrometry is a versatile basis to analyse greenhouse gases in the atmosphere. A multicomponent air pollution software (MAPS) was developed for retrieval of gas concentrations from radiation emission as well as absorption measurements. Concentrations of CO, CH4, N2O, and H2O as well as CO2, NO, NO2, NH3, SO2, HCl, HCHO, and the temperature of warm gases are determined on-line. The analyses of greenhouse gases in gaseous emission sources and in ambient air are performed by a mobile remote sensing system using the double-pendulum interferometer K300 of the Munich company Kayser-Threde. Passive radiation measurements are performed to retrieve CO, N2O, and H2O as well as CO2, NO, SO2, and HCl concentrations in smoke stack effluents of thermal power plants and municipal incinerators and CO and H2O as well as CO2 and NO in exhausts of aircraft engines. Open-path radiation measurements are used to determine greenhouse gas concentrations at different ambient air conditions and greenhouse gas emission rates of diffusive sources as garbage deposits, open coal mining, stock farming together with additional compounds (e.g. NH3), and from road traffic together with HCHO. Some results of measurements are shown. A future task is the verification of emission cadastres by these inspection measurements.

It is essential to establish an objective and quantitative method for evaluating facial palsy and to measure the extent of paralysis in order to evaluate therapeutic efficacy, determine prognosis, select appropriate treatment and observe the process of recovery. This study utilized Moiré topography, which displays three-dimensional facial symmetry with high precision and is based on light interference theory, to determine the extent of facial palsy in 38 patients (20 men and 18 women) 5 months to 73 years of age. A stereoscopic lattice type Moiré camera (FM3013) was connected to a CCD camera and to the monitoring device for confirming Moiré stripes. Moiré photographs were taken with a thermal imager (FTI-200). The photos were visually and objectively evaluated on the basis of the Moiré pattern and were then input into a personal computer with a digitizer for data processing and analysis. To view the functions of facial nerve branches, five Moiré photographs were taken: at rest, wrinkling the forehead, closing the eyes lightly, blowing out the cheeks and grinning. Results indicated that the number of stripes and their polarization adequately reflected the function of individual facial nerve branches. Thus, a well-defined Moiré pattern could clarify the characteristics of the site and the degree of facial palsy and of recovery from paralysis. It is an analytical method that can be quickly applied and seems especially useful in infants and young children, in whom point-based assessment is difficult. It is possible to quantitatively evaluate facial palsy in terms of the Asymmetry Index (AI), which is 20-25% for severe paralysis, 12-19% for partial paralysis, and 5-10% for an essentially normal condition. However, the numerical value of the AI overlap in all three paralysis categories, indicating that quantitative assessment of paralysis would be difficult. Moiré topography is an excellent method of determining the extent of facial palsy, compensating for the short falls of examination-based assessment and permitting reproducible visual, objective, and quantitative evaluation.

BACKGROUND

Farnesyl protein transferase inhibitors have emerged as promising novel agents for combating cancerous disease. Nevertheless, the importance for farnesyl protein transferase enzymatic activity for cellular physiology of untransformed cells remains poorly investigated.

METHODS

Peripheral blood monocytes, isolated from the blood of eight healthy volunteers, were treated with a farnesyl protein transferase inhibitor (FTI 744,832) or vehicle control for 16 hr. Subsequently cells were challenged with different concentrations of lipopolysaccharide (LPS), colony stimulating factor-1 (CSF-1), or phorbol esters for 10 min, after which the activation state of p42/p44 MAP kinase, p38 MAP kinase, and Jun-N-terminal kinase was investigated using Western blotting and phosphospecific antibodies.

RESULTS

We observed that farnesyl protein transferase inhibition abrogated activation of p38 MAP kinase by LPS, CSF-1, and phorbol esters. Also the activation of Jun-N-terminal kinase by LPS was not seen after farnesyl protein transferase inhibition. Finally, stimulation of p42/p44 MAP kinase with CSF-1 was strongly reduced by farnesyl protein transferase inhibition, whereas activation of p42/p44 MAP kinase by phorbol ester was only slightly effected.

CONCLUSIONS

Farnesyl protein transferase enzymatic activity is required for proper activation of all major members of the MAP kinase family. The observation that activation the p38 MAP kinase and Jun-N-terminal kinase is sensitive to farnesyl protein transferase inhibition raises the possibility that, in addition to cancerous disease, farnesyl protein transferase inhibitors may be useful compounds in combating inflammatory disease.

The aim of this study was to explore the feasibility of the family talk intervention (FTI) and its acceptability to dependent children when a parent is cared for in palliative home care. The main goal of FTI is to increase family communication about the illness. The present paper derives from a pilot study and is based on 25 children's reports, involving both questionnaires and interviews, after participation. A majority of the children appreciated the structure and content of FTI. They felt seen, heard, and acknowledged by the interventionists and recommended FTI to other children in similar situations.

Thyroxin (T4) and triiodothyronine (T3) were measured by radioimmunoassay in serum of hamsters sacrificed at 4-hr intervals throughout the daily light-dark cycle (14L/10D). Both T4 and T3 concentrations increased significantly during the L period of the daily cycle and decreased during the D period of the cycle; A.M. versus P.M. differences in free thyroxin indices (FTI) were also studied using the T4 and T3 uptake assays of Nuclear Medical Laboratories (Dallas, Texas). The free thyroxin index was significantly greater in serum samples of hamsters sacrificed at 7 P.M. than at 7 A.M. (lights on at 6:30 A.M.). Serum taken at 7 P.M. had less unsaturated binding sites than serum taken at 7 A.M. No significant A.M. versus P.M. differences in free thyroxin index were found in blind hamsters, although blind hamsters had significantly lower T4 and FTI than controls. Placing melatonin in the drinking water at a dose of 80 micrograms/ml did not significantly influence hormone levels. The greatest difference in hormone concentrations between control and blinded hamsters was found in P.M. samples. Blind hamsters had FTIs that were 48% of P.M. controls. Pinealectomy prevented the effects of blinding on T4 levels and FTIs.

The first synthesis of the tetracyclic aromatic compound furo[2,3- g]thieno[2,3- e]indole ("FTI") is described. The synthetic strategy features a photochemical benzannulation based on the reaction of an α-diazo ketone and ynamide which assembles a benzothiophene equipped with substituents that enable subsequent cyclizations to generate the nitrogen and oxygen heterocyclic rings.

Epidermal growth factor (EGF) is an important mitogen and its secretion in neonatal animals has been shown to be affected by thyroid hormone levels. EGF in blood of humans is found in both platelets (as reflected in its serum level) and in plasma; its origin in plasma remains unclear. Serum and plasma EGF were studied in a group of patients with thyroid disorders. Twenty hyperthyroid subjects (3M, 17F) aged 37.3 +/- 14.9 years and 10 hypothyroid patients (3M, 7F) aged 58.3 +/- 18.6 years were studied before and after euthyroidism was restored. Before treatment, serum EGF in the hyperthyroid patients was elevated compared to normal controls (501 +/- 376 vs 270 +/- 154 pmol/l, p less than 0.001). After treatment of hyperthyroidism, serum EGF returned to the normal levels (232 +/- 176 pmol/l). In contrast, serum EGF was not significantly different in the hypothyroid subjects either before or after treatment (151 +/- 194 and 237 +/- 153 pmol/l respectively). A significant correlation (r = 0.461, p less than 0.001) between serum EGF and serum-free thyroxine index (FTI) was found when all samples from both untreated and treated hyper- and hypothyroid patients were examined. Multiple regression analysis revealed that both serum FTI and platelet count independently affected the serum EGF levels. Similarly, plasma EGF was also elevated in untreated hyperthyroid patients with a median of 26.4 pmol/l (range less than 16.6-88.0), whereas all normal controls and hypothyroid subjects had unmeasurable levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Thyroid function was investigated in 17 patients with acute intermittent porphyria (AIP). In 13 patients in remission and latent for the disease, thyroid function was normal. In contrast, transient hyperthyoridism with elevated total T4, T3 free T4 and T3, and FTI levels occurred in one pregnant patient with severe attacks of AIP, and similar, though less pronounced, alterations in thyroid function were found in three other symptomatic female patients. It is suggested that increased thyroidal sympathetic neural stimulation is responsible for these changes in thyroid function.

We have described a 25-year-old man with hyperthyroidism and normal levels of serum T3 and reverse T3, but with abnormalities in serum T4I, T3U, FTI, FT4, and T3 suppression and TRH stimulation tests consistent with the diagnosis of T4 toxicosis. Serum total iodine was elevated several-fold, and the observed response to iodine depletion and loading suggested that the T4 toxicosis in this patient probably resulted from an increased iodine load, with production of T4 preferentially to T3. There appeared to be no evidence of a defect in peripheral conversion of T4 to T3.

To determine if a sensitive assay for thyrotropin (thyroid-stimulating hormone [TSH] ) would be useful as a primary test to determine optimal doses of levothyroxine, an enzyme immunoassay for TSH that discriminated hyperthyroid from euthyroid subjects with no overlap was compared with radioimmunoassays for TSH and thyroxine (T4) and the calculated free thyroxine index (FTI) in 100 patients receiving stable doses of levothyroxine. The basal TSH level, determined with the sensitive assay, predicted the TSH response to thyrotropin-releasing hormone (TRH); all patients whose basal TSH level was less than 0.35 mU/L had absent or subnormal responses, and all patients with TSH levels of 0.35 mU/L or greater had normal responses. In patients with normal TSH levels, the T4 level and FTI were superfluous (normal) or misleadingly high; conversely, 65% of patients with low TSH levels had normal T4 levels or FTIs, or both. An assay for TSH with sufficient sensitivity to discriminate between abnormally low values and the low range of normal can be used as the primary test for monitoring levothyroxine treatment. The T4 or FTI measurements are not needed if the TSH level is normal, but they should be done in patients with low TSH levels.

The Amerlex FT4 radioimmunoassay has been assessed. It is simple, quick, and precise. The assay is as good as the FTI or the Immo Phase FT4 assay in hyperthyroid, hypothyroid, or elderly subjects and in women on oral contraceptives. In acute illness and pregnancy, however, over half the values lie below the euthyroid reference range. Further work on blood samples obtained from pregnant subjects demonstrated that while FT4 values given by the Immo Phase and Liquisol kits remain constant, those given by the GammaCoat, Amerlex, and Lisophase kits show a significant decrease with gestation. What the 'true' FT4 is in acute illness and in pregnancy remains an open question.

The correlation of the diagnostic indices FTI and T4/TBG with actual free T4 levels (FT4) was investigated by numerical analysis. It was shown that, FT4 being kept constant, both indices vary with altered TBG concentrations. Thus neither index is truly proportional to FT4. In this respect both indices appear doubtful tools for diagnosing patients with abnormal TBG levels. A "map" constructed by plotting T4 versus T3U appeared to offer a higher discriminatory potential with respect to pathologic FT4 values. Such an idealized map was shown to consist of hypo-, eu- and hyperthyroid domains separated by lines governed by the limit values of the FT4 normal range. In practice, of course, these domains are separated by border regions rather than border lines and they should be derived from clinical data.

The unstimulated plasma thyroid indices of triiodothyronine uptake (T(3)U), thyroxine (T(4)), free thyroxine index (FTI), thyroid-stimulating hormone (TSH), and prolactin levels were measured in 56 patients who were complaining of secondary amenorrhea, and in 40 control patients. No significant differences were noted in the plasma indices between the two groups. Mean ± standard error plasma prolactin levels in patients with secondary amenorrhea and in the control patients were 21.8 ± 2 ng/mL and 10.5 ng/mL, respectively. Twenty-two (39 percent) patients who were complaining of amenorrhea had hyperprolactinemia and galactorrhea. The thyroid indices of these patients did not differ significantly from the control group.

An increased incidence of cold-reactive lymphocytotoxic activity (LCTA) has been demonstrated in the sera of patients with autoimmune thyroid disease. Twenty-six of 79 (33%) patients with Graves' disease and 9 of 21 (43%) patients with Hashimoto's thyroiditis had cold-reactive LCTA detected by microcytotoxicity assay compared to 6 of 42 (14%) normal controls. There was no correlation between LCTA and age, sex, MCHA titre or TGHA titre. A positive correlation with FTI and LCTA in Hashimoto's patients was demonstrated, but no such correlation was demonstrable in Graves' patients. The lymphocytotoxic activity was directed preferentially against B cells. There was no preferential lysis of T-cell subsets as defined by monoclonal antibodies, and the lymphocytotoxins were equally reactive with normal lymphocytes and toxic Graves' lymphocytes. The significance of cold-reactive lymphocytotoxic activity in the pathogenesis of autoimmune thyroid disease remains to be determined.

Cells sense physical properties of their extracellular environment and translate them into biochemical signals. In this study, cell responses to surfaces with submicron topographies were investigated in cultured human NF1 haploinsufficient fibroblasts. Age-matched fibroblasts from 8 patients with neurofibromatosis type 1 (NF1(+/-)) and 9 controls (NF1(+/+)) were cultured on surfaces with grooves of 200 nm height and lateral distance of 2 μm. As cellular response indicator, the mean cell orientation along microstructured grooves was systematically examined. The tested NF1 haploinsufficient fibroblasts were significantly less affected by the topography than those from healthy donors. Incubation of the NF1(+/-) fibroblasts with the farnesyltransferase inhibitor FTI-277 and other inhibitors of the neurofibromin pathway ameliorates significantly the cell orientation. These data indicate that NF1 haploinsufficiency results in an altered response to specific surface topography in fibroblasts. We suggest a new function of neurofibromin in the sensoric mechanism to topographies and a partial mechanosensoric blindness by NF1 haploinsufficiency.

Background: Plantar pressure distribution after the first metatarsal proximal crescentic osteotomy (FMPCO) with lesser metatarsal proximal shortening osteotomy (LMPSO) for hallux valgus with metatarsalgia has not been previously described.

Methods: The pre- (Pre) and postoperative (Post) groups comprised of 18 patients who underwent unilateral FMPCO with LMPSO; fifteen healthy volunteers constituted the control (C) group. For each of the 10 regions, peak pressure (Peak-P), maximum force (Max-F), contact time (Con-T), contact area (Con-A), and force-time integral (FTI) were measured.

Results: The mean Peak-P of the second metatarsal head was significantly lower in the Post group than the Pre group. The mean Peak-P, Max-F, Con-T, and FTI were not significantly different between the Post and C groups. The mean Con-A was significantly lower in the Post group than the C group.

Conclusion: FMPCO with LMPSO may improve the plantar pressure of the central forefoot comparable to healthy subjects.

Keywords: Hallux valgus; Metatarsalgia; Osteotomy; Plantar pressure.

It is still a challenge to delay the onset of fatigue on muscle contraction induced by Functional Electrical Stimulation (FES). We explored the use of two stimulation methods with the same total area, single electrode stimulation (SES), and spatially distributed electrical stimulation (SDSS) during isometric knee extension with spinal cord injured (SCI) volunteers. We applied stimulation on the left and right quadriceps of two SCI participants with both methods and recorded isometric force and evoked electromyography (eEMG). We calculated the force-time integral (FTI) and eEMG-time integral (eTI) for each stimulation series and used a linear regression as a measure of decay ratio. Moreover, we also estimated the contribution from each channel from eEMG.

In a randomised controlled study in 16 orthopaedic patients, the influence of midazolam-fentanyl-N2O/O2 anesthesia (group A) resp. halothane-N2O/O2 anesthesia (group B) on the plasma concentrations of the endocrine parameters ACTH, aldosterone, cortisol, 17-DHEA, insulin, prolactin, T3, T4, TBG (thyroxine bounded globuline) as well as adrenaline, noradrenaline, and dopamine was investigated. Additionally the metabolites glucose, lactate, free glycerin, and acetacetate were measured. Beside prolactin values, only the values for ACTH, aldosterone, cortisol, and 17-DHEA differed with respect to both anesthesia methods. Under halothane-N2O/O2 anesthesia free T4 rose initially also, here represented by T4/TBG-ratio (= FTI). However, the fall of T3 concentration showed no phase - resp. anesthesia-specific changes. Catecholamine levels reached highest values towards the end of operation resp. one hour after extubation in both groups. The insulin secretion, however, was not significantly raised in either group during acute stress phases. As an expression of modified metabolic regulation comparable rises of plasma levels of glucose, lactate, free glycerin, and acetacetate were observed under midazolam-fentanyl-N2O/O2 anesthesia as well as under halothane-N2O/O2. According to presented data, both methods of anesthesia modulated the endocrine metabolic response of the organism to surgical stress, without showing any clinically relevant advantages or disadvantages attributable to either method.

Thyroid function was examined periodically in a group of 85 patients treated with low-dose amiodarone, and followed for 1-13 years (mean 3.6). Biochemical hypothyroidism (elevated thyroid stimulating hormone (TSH) only) developed in 8 patients and clinical hypothyroidism in 3, while hypothyroidism developed in 5. In the first 11 cases amiodarone was continued but 1-thyroxine was added. In the 5 that became hyperthyroid, amiodarone was stopped and thyroid function became normal within a few months. In the 69 patients without thyroid dysfunction, elevated thyroxine (T4) and free thyroxine index (FTI) were found in 20% and 17%, respectively, and elevated free T4 (FT4) in 28%; all had normal total tri-iodothyronine (TT3) and TSH. In practically all patients, reverse T3 (rT3) rose more than 30% above initial levels. It is concluded that in patients treated with low dose amiodarone: 1) thyroid function should be followed closely; 2) elevated levels of TT4, FTI and FT4 do not necessarily imply hyperthyroidism; 3) hypothyroidism may be biochemical only, without progression to overt hypothyroidism, despite continued treatment; 4) both the hypo- and the hyperthyroidism induced are usually reversible; 5) amiodarone may be continued despite onset of hypothyroidism, provided replacement therapy is given.

Farnesyl:protein transferase (FPTase) catalyzes the covalent addition of the isoprenyl moiety of farnesylpyrophosphate to the C-terminus of the Ras oncoprotein and other cellular proteins. Inhibitors of FPTase (FTIs) have been developed as potential anticancer agents, and several compounds have been evaluated in clinical trials. To facilitate the identification of cell-active FTIs with high potency, the authors developed a method that uses a radiolabeled FTI that serves as a ligand in competitive displacement assays. Using high-affinity [(3)H]-labeled or [(125)I]-labeled FTI radioligands, they show that specific binding to FPTase can be detected in intact cells. Binding of these labeled FTI radioligands can be competed with a variety of structurally diverse FTIs, and the authors show that inhibition of FTI radioligand binding correlates well with inhibition of FPTase substrate prenylation in cells. This method provides a rapid and quantitative means of assessing FTI potency in cells and is useful for guiding the discovery of potent, novel inhibitors of FPTase. Similar methods could be employed in the optimization of inhibitors for other intracellular drug targets.