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At least 451 records · Page 25Linked to original sources

Combination chemotherapy for metastatic breast cancer with fluorouracil, adriamycin, cyclophosphamide, and methotrexate.

Twenty-nine patients with metastatic breast cancer were treated with fluorouracil, adriamycin, cyclophosphamide (FAC), and methotrexate (MTX), with or without leukovorin rescue. Of 24 evaluable patients, one achieved a complete remission and 17 had partial responses. The overall objective response rate was 75%. The median survival from initiation of chemotherapy for the responding patients was 18 months. Four patients (17%) with stable disease had a median survival of 25 months. The addition of MTX to FAC chemotherapy did not improve the therapeutic efficacy of this combination; it did, however, increase the overall toxicity, especially the infectious complications when compared to FAC alone.

Alopecia↗

Determination of interferon-gamma in Chinese hamster ovary cell culture supernatant by coupled-column liquid chromatography.

A robust tandem HPLC method coupling size-exclusion (Shodex Asahipak GS-320HQ) and reversed phase (Vydac 218TP54) columns with ultraviolet detection was developed for quantitative determination of interferon-gamma (IFN-gamma) in Chinese hamster ovary cell culture supernatant. The 2D-HPLC system was linked up by a 6-port 2-position low hold-up volume switch valve. Compared to a commercial ELISA kit for IFN-gamma, the coupled column LC approach was able to detect and quantify soluble IFN-gamma, regardless of the glycoprotein's molecular/conformational variability and sample background. Each LC-LC analysis took 90 minutes inclusive of column regeneration. The relative standard deviation of measurements (n = 5) was less than 3%. The limit of detection (LOD) was determined to be 0.35 microg IFN-gamma.

Animals↗

2-Octylcynanoacrylate-assisted microvascular anastomosis in a rat model: long-term biomechanical properties and histological changes.

The aim of this study was to establish the long-term biomechanical and histological properties of 2-octylcyanoacrylate-assisted microvascular anastomosis over conventional suture-only anastomosis in the laboratory rat model. The biomechanical and histological properties of three groups of vessels were compared: 1) vessels with 2-octylcyanoacrylate-assisted anastomoses (study group); 2) vessels with suture-only anastomoses (control group); and 3) normal unoperated vessels (sham group). In total, 144 adult rats were used, and these were studied at 1 week, 1 month, 3 months, and 6 months postanastomosis. At 6 months, the tensile strength of study vessels was significantly higher than control vessels. The stiffness of study and control vessels was similar at all time intervals. Histologically, there was no evidence that 2- octylcyanoacrylate caused toxicity to vessel walls, and there was less perivasacular foreign-body giant-cell reaction in the study group compared to the control group. Long-term follow-up showed that microvascular anastomosis with 2-octylcyanoacrylate in rat femoral arteries had superior tensile strength and similar stiffness to vessels anastomosed with sutures only, without adverse effects to surrounding tissues.

Anastomosis, Surgical↗

The tautomerism of 1H-pyrazole-3(5)-(N-tert-butyl)carboxamide in the solid state and in solution.

The X-ray crystal structure of 1H-pyrazole-3-(N-tert-butyl)-carboxamide was determined. In the solid state, the 13C and 15N CP/MAS NMR spectra correspond to this tautomer. In solution, both tautomers are present in a ratio that depends on the temperature (at 293 K, 90% 3-substituted/10% 5-substituted). Some unusual 1H, 1H couplings involving the NH proton were observed. DFT (GIAO) calculations were carried out.

Journal Article↗

Effects of velocity on maximal torque production in poststroke hemiparesis.

Impaired torque production is a major physical impairment following stroke, and has been studied extensively in isometric conditions. However, functional use of a limb requires torque production during movement, and the effects of velocity on maximal torque production may be abnormally enhanced in the paretic limb. The purpose of this study was to quantify the effects of movement velocity on maximal torque production during isokinetic, concentric flexion and extension of the elbow in poststroke subjects. Three speeds were tested (30, 75, 120 deg/s) over a 100-deg range of motion. To control for strength variations between subjects and limbs, isokinetic torques were normalized by peak isometric torque. As flexion velocity increased, paretic limb torque decreased at a greater rate than in the unaffected limb. During extension, paretic limb torque was much lower than torque in the unaffected limb at all speeds. In both flexion and extension, the disparity between limbs in the constant-velocity torque-angle curves became more pronounced as velocity increased. Torque decreased 44% +/- 7% in flexion and 63% +/- 9% in extension as velocity increased from 30 to 120 deg/s, whereas the corresponding decreases in the unaffected limb were only 9% +/- 5% in flexion and 16% +/- 4% in extension. No electromyographic (EMG) abnormalities were observed during flexion. During extension, EMG data provided evidence for abnormally increased antagonist coactivation in brachioradialis and markedly reduced activation in triceps as potential contributors to the decreased extension torques. The finding that movement velocity produces large deficits in maximal torque might explain why functional use of the paretic limb is often impaired even though isometric strength appears adequate.

Aged↗

New muscle transplant method produces normal twitch tension in dystrophic muscle.

Grafting newborn muscle is an innovative method of muscle transplant. This method overcomes hypoxia in the deeper fibers and facilitates reinnervation and revascularization of the grafted muscle fibers, thus promoting the survival and development of the characteristics of the donor muscle. The result achieved is superior to that obtained from mature muscle grafts or from minced muscle transplants. When an intact soleus from a 1-day-old normal mouse was grafted into a recipient soleus of a 20-day-old dystrophic C57BL/6J-dy2J mouse, the actively developing normal graft helped to improve the structure and function of the dystrophic muscle. When compared to the intact dystrophic solei, the test dystrophic muscles five to six months after operation showed increases in cross-sectional area, in wet weight, in twitch and tetanic tension, and in the number of muscle fibers with high resting membrane potentials. This is the first procedure to have raised the muscle twitch tension in an adult dystrophic mouse to the normal level.

Action Potentials↗

p53 expression, mutation, and allelic deletion in ovarian cancer.

The aim of this investigation was to study the prevalence of p53 gene mutations and allelic deletions in ovarian cancer and the relationship between these events and p53 expression. Genomic p53 was amplified by the polymerase chain reaction (PCR) from paraffin-embedded sections of tumour and non-tumour tissue. Exons 5-9 were screened for mutation using non-isotopic single-strand conformation polymorphism (SSCP) analysis. Allelic deletions at a sequence polymorphism in exon 4 were studied for loss of heterozygosity (LOH) by restriction analysis and by SSCP. p53 expression was detected by immunohistochemistry with the p53 antibodies CM1 and Do7. Of 44 cases, ten (23 per cent), including a mucinous tumour of low malignant potential, showed mutations. There was a statistically significant correlation between p53 mutation and expression (P < 0.01) but seven cases showed discordance. Of 18 informative cases, eight (44 per cent) demonstrated LOH. Mutations were identified in three of the informative cases, two of which also had LOH. The remaining case showed mutations in two amplicons. p53 dysfunction, as indicated by mutation, deletion, or increased expression, is common in ovarian cancer. There is an association between these molecular events but the exact mechanisms of p53 inactivation remain to be elucidated.

Alleles↗

Detection of loss of heterozygosity of p53 gene in paraffin-embedded breast cancers by non-isotopic PCR-SSCP.

A rapid non-isotopic PCR-SSCP (polymerase chain reaction-single-stranded conformation polymorphism) method was developed in this study to detect polymorphism and loss of heterozygosity (LOH) of p53 in formalin-fixed and paraffin-embedded samples of normal breast tissue and of breast cancer. p53 expression was also examined by immunohistochemistry. In 35 paired samples, heterozygosity in exon 4 of p53 was detected in 17 cases (49 per cent) and LOH of the p53 gene in breast cancer tissues was observed in 7 out of 15 informative cases (47 per cent). The correlation of LOH of p53 with positive p53 immunostaining did not reach statistical significance, but all immunostaining-positive tumours among informative cases had LOH of p53. The results support the hypothesis that in most cases the allelic deletion of p53 may uncover the 'recessive mutation' in the remaining allele. However, LOH of p53 was more frequent than positive immunostaining and was significantly associated with poor differentiation of breast cancer (P < 0.05). The results suggest that the allelic deletion of p53 may also contribute to the development and progression of breast cancer by reducing the amount of normal p53 protein. These results show that non-isotopic PCR-SSCP is a simple, fast, and effective method for detecting polymorphism and LOH of the p53 gene, which is especially useful for retrospective studies.

Adult↗

The short-chain homologue of dihydrolipoic acid, tetranordihydrolipoate, protects against iron-induced lipid peroxidation in the aqueous phase.

Because iron is involved in catalysis of many biological oxidations, it is important to investigate new and novel antioxidants in terms of their effect on iron-catalyzed oxidations. We investigated the effect of dihydrolipoic acid (6,8-dimercaptooctanoic acid (DHLA)), its homologues (4,6-dimercaptohexanoic acid (bisnorDHLA) and 2,4-dimercaptobutanoic acid (tetranorDHLA)) and methyl 6,8-dimercaptooctanoate (methylDHLA) on Fe(II)-citrate-catalyzed lipid peroxide-dependent lipid peroxidation in lipid-dispersed and liposome systems. In the lipid-dispersed system, tetranorDHLA inhibited conjugated diene formation induced by Fe(II)-citrate. In the presence of tetranorDHLA, oxygen was consumed more rapidly in the reaction mixture than in the presence of the other compounds, but the oxidation rate of Fe(II)-citrate in the reaction mixture was slower than in the presence of the other compounds. This suggests that tetranorDHLA inhibited lipid peroxidation by the oxidation of tetranorDHLA itself at the site where the lipid was oxidized.

In Vitro Techniques↗

WWP, a new amino acid motif present in single or multiple copies in various proteins including dystrophin and the SH3-binding Yes-associated protein YAP65.

A new repeating amino acid motif, which we called WWP, was found in several proteins of yeast, nematod or vertebrate origin. Among these are dystrophin, the product of the Duchenne muscular dystrophy locus, a protein (YAP65) which associates in vitro with the Src homology domain 3 (SH3) of the Yes proto-oncogene product, and a human putative GTPase-activating protein. As is the case for proteins which contain the SH2, SH3 and PH domains, at least some of the WWP-containing proteins appear to be signaling or cytoskeletal proteins, associated with plasma or organellar membranes, and specific protein-protein contacts are likely to be crucial to their function.

Adaptor Proteins, Signal Transducing↗

Arachidonic acid activation of monocyte and neutrophil reactive oxygen in lung cancer patients undergoing pulmonary resection.

Reactive oxygen intermediate (roi) generation was investigated in phagocytes of 39 patients undergoing pulmonary resection for lung cancer and 39 paired healthy controls. Generation of roi in monocytes and neutrophils was monitored using 2',7'-dichlorofluorescin diacetate. Activation associated with hydrophobic interactions was probed by analysis of phagocyte roi activation by arachidonic acid and gamma-linolenic acid. Patient roi was measured pre-operatively and 2 and 7 days post-operatively. Elevated (P < 0.01) roi production was detected in neutrophils of lung cancer patients. Surgery was associated with an increase (P < 0.05-P < 0.01) in phagocyte roi at 2 and 7 days post-op. Phagocyte roi was stimulated by arachidonic acid and gamma-linolenic acid (1-40 microM) both pre- and post-operatively. Differences in arachidonic acid and gamma-linolenic acid stimulation between patient and control and pre- and post-op patient phagocytes suggest arachidonic acid involvement in phagocyte activation during reactive responses to lung carcinoma and surgery.

Arachidonic Acid↗

Ligation of CD40 potentiates Fas-mediated activation of the cysteine protease CPP32, cleavage of its death substrate PARP, and apoptosis in Ramos-Burkitt lymphoma B cells.

The proliferation and survival of a B cell population is necessarily tightly controlled to prevent the arisal of malignancy or autoimmunity. The expansion or elimination of a B cell clone is determined through a complex interaction of the tumour necrosis factor receptor/nerve growth factor receptor family members CD40 and Fas, which are expressed on the B cell surface, with their respective physiological ligands (CD40L and FasL) expressed on the surface of CD4+ T cells. The regulation of B cell growth by signals transduced through CD40 and Fas contributes to the maintenance of peripheral tolerance and likely takes place and in the germinal centres (GC) of secondary lymphoid tissues. In this study, we investigate the relationship between the expression of Fas and B cell survival following engagement of CD40 and Fas in the Epstein-Barr virus-genome-negative Ramos-Burkitt lymphoma (Ramos-BL) B cell line model of GC B lymphocyte selection during maturation of the humoral immune response. We now present evidence that Ramos-BL B cells constitutively express both Fas and FasL on their surface and that expression of Fas, but not FasL, is enhanced following ligation of CD40. Coligation of CD40 and Fas, triggers for growth inhibition, activation of the interleukin-1 beta-converting enzyme, now caspase, family member CPP32 (caspase-3) but not Ich-1L (caspase-2), cleavage of its death substrate poly(ADP-ribose) polymerase, and apoptosis from the G1 phase of cell cycle; engagement of Fas alone fails to trigger for growth inhibition and apoptosis but enhances AgR-mediated cellular death. This CD40-potentiated Fas-triggered growth inhibition and apoptosis occurs in the presence of CD40-induced expression of the anti-apoptotic proteins Bcl-xL and Bcl-2. Taken together, these data indicate that ligation of CD40 facilitates efficient coupling of Fas to the caspase-mediated pathway of apoptosis.

Antibodies, Monoclonal↗

Modulation of MHC expression on human endothelial cells by sera from patients with systemic lupus erythematosus.

Major histocompatibility complex (MHC) antigen expression on cells is a prerequisite for immune interaction with activated T-cells. This study examined the ability of sera from patients with systemic lupus erythematosus (SLE) to modulate MHC expression on vascular endothelial cells. SLE sera were able to selectively upregulate MHC class I antigen expression on cultured human umbilical venous endothelial (HUVE) cells, without concomitant induction of MHC class II antigen. The stimulation index (SI) for MHC class I expression produced by SLE sera (1.21 +/- 0.23) was significantly higher than those for normal controls (1.01 +/- 0.10) (P < 0.0001) and non-SLE patients (1.12 +/- 0.14) (P < 0.05). Additionally, active SLE patients had higher mean SI than inactive patients (P < 0.001). Preincubation of SLE sera with Protein A-Sepharose beads conjugated with antibodies against tumor necrosis factor-alpha and interferon-alpha was able to significantly reduce their ability to upregulate class I MHC expression by HUVE cells, indicating that these cytokines were responsible for the modulatory effect. This could be an important mechanism for the immune-mediated vascular injury seen in SLE.

Adolescent↗

Contact inhibition of cell spreading: a mechanism for the maintenance of thyroid cell aggregation in vitro.

When freshly isolated porcine thyroid cells are stimulated with thyrotropin (TSH) they organize to form functional follicles in conventional substrate-adherent culture. Cell aggregation is essential for follicular reorganization and is likely to be influenced by the balance between cell-cell adhesion (promoting aggregation) and cell-substrate adhesion (favoring spreading and monolayer formation). Recently we observed that TSH potentiated cell-cell adhesion and in the present study we have sought evidence that TSH might also regulate cell-substrate adhesion. Two parameters of cell-substrate adhesion, namely, cell attachment to collagen and cell spreading upon collagen, were measured using preparations of isolated single cells and of multicellular aggregates. TSH had no effect upon the attachment or spreading of single cells, but inhibited aggregate spreading without affecting aggregate attachment. The possibility that cell-cell contact modulated the response to TSH in aggregates, but not in single cells, was confirmed using a cell-free membrane preparation which inhibited the spreading of single cells but not their rate of attachment. Moreover, TSH potentiated the inhibitory effect of membranes on the spreading of single cells. Heparin also specifically inhibited the spreading of both single cells and cell aggregates, suggesting that a heparin-sensitive adhesive mechanism might be recruited as thyroid cells spread. We conclude that thyroid cell-substrate adhesion is regulated by a synergistic interaction between cell-cell contact and TSH which preferentially inhibited cell spreading but not attachment. Such contact-dependent inhibition of cell spreading is predicted to preserve cell aggregates and hence contribute to the maintenance of thyroid follicular differentiation in vitro.

Animals↗

Thyroid cell spreading and focal adhesion formation depend upon protein tyrosine phosphorylation and actin microfilaments.

Adhesion to proteins of the extracellular matrix exerts a profound influence upon cell function and behavior. Similar adhesive interactions mediate the spreading of cultured cells upon artificial substrata. Recently we observed that thyrotropin (TSH) and intercellular contact regulated thyroid cell-substrate adhesion to inhibit cell spreading, but not initial attachment. This is a mechanism which preserves thyroid follicular differentiation in culture. In the present study we have investigated the role of cytoplasmic components in mediating thyroid cell adhesion to collagen. The earliest change associated with cell spreading was the accumulation of vinculin and phosphotyrosine in developing focal adhesions, which was followed by stress fiber and microtubule assembly. Genistein, an inhibitor of tyrosine kinases, and cytochalasin B inhibited cell spreading and focal adhesion formation without affecting initial attachment to substrate. In contrast microtubule disorganization by colchicine did not alter any parameter of thyroid cell-substrate adhesion. These observations indicate that protein tyrosine phosphorylation and dynamic microfilament integrity are essential for attached thyroid cells to spread upon substrate. They are therefore potential intracellular loci at which TSH and intercellular contact may regulate cell adhesion to extracellular matrix and influence thyroid cell behavior.

Actin Cytoskeleton↗

Thyroid epithelial morphogenesis in vitro: a role for bumetanide-sensitive Cl- secretion during follicular lumen development.

The structural and functional unit of the thyroid gland is the follicle, consisting of a closed lumen surrounded by a single layer of polarized epithelial cells. In this paper we have attempted to characterize the process of lumenal development when primary cultures of porcine thyroid cells reorganized to form follicles. Cells incubated with the loop diuretic, bumetanide, an inhibitor of NaK2Cl cotransport, aggregated but failed to form normal follicles. Laser scanning confocal microscopy combined with immunohistochemical markers of thyroid cell-surface proteins demonstrated that in the presence of bumetanide cells polarized and assembled ZO-1-containing tight junctions separating their apical and basolateral membrane domains. Cultures formed small lumena but their subsequent growth was inhibited by bumetanide. Electrophysiological studies confirmed that bumetanide-sensitive Cl- transport was the major contributor to the transepithelial electrical potential difference across the follicular wall after 48 h incubation. Other potential mechanisms did not contribute significantly to follicular lumenal growth. In particular, bumetanide did not affect cell proliferation and, in contrast to tissue follicles, thyroglobulin could not be detected within the lumena of cultured follicles. We conclude that thyroid follicular reorganization involves two distinct and separate phases of lumenal development: initial lumen formation which probably reflects the assembly of a specialized apical membrane domain; and subsequent lumenal growth which is mediated by the inward transport of Cl- by polarized epithelial cells.

Animals↗