Biomedical subjects

D J Beech

Publications and source records attributed to D J Beech.

82 records · Page 5Linked to original sources

PubMed1987

Voltage dependence of the actions of nifedipine and BAY K 8644 on barium currents recorded from single smooth muscle cells from the rabbit ear artery.

The voltage dependence of the actions of nifedipine and BAY K 8644 were studied on whole-cell barium currents recorded from single, collagenase- and elastase- dispersed, smooth muscle cells from the rabbit ear artery using standard patch clamp techniques (Hamill et al., 1981). The results showed that nifedipine (3 x 10(-7)M) reduced the inward current in response to depolarizations positive of -10mV but in the threshold range, -40mV to -20mV, that it increased the amplitude of the peak barium current. Subsequent application of BAY K 8644 (1.5 x 10(-7)M) enhanced this latter effect but decreased the inhibition induced by nifedipine at more positive potentials.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1987

The effects of tetraethylammonium ions, 4-aminopyridine or quinidine on K+-currents in single smooth muscle cells of the rabbit portal vein.

Whole-cell and isolated patch configurations of the patch clamp technique were used to distinguish K+-channels by their pharmacological characteristics in single, enzymatically dispersed, smooth muscle cells from the rabbit portal vein. Results suggest that spontaneous transient outward currents are carried mainly by large conductance Ca2+-activated K+-channels and outward current evoked by depolarization of the membrane is carried partially by these channels but also by a time- and potential-dependent K+-channel blocked by 4-aminopyridine.

4-Aminopyridine↗

PubMed1985

Electrophysiological and other aspects of the relaxant action of isoprenaline in guinea-pig isolated trachealis.

In guinea-pig isolated trachealis isoprenaline (0.001-0.1 mumol l-1) caused concentration-dependent relaxation. Propranolol (1 mumol l-1) antagonized the effects of isoprenaline by more than 100 fold but did not modify the relaxant action of sodium nitrite. The tracheal relaxant actions of isoprenaline and ATP were unaffected by apamin (0.1 mumol l-1) but apamin profoundly antagonized the effects of noradrenaline and ATP on guinea-pig isolated taenia caeci. Tetraethylammonium (TEA; 8 mmol l-1) and procaine (5 mmol l-1) each evoked tracheal spasm but neither agent antagonized the isoprenaline-evoked relaxation of the trachealis. Trachealis exposed to K+-rich (120 mmol l-1) Krebs solution developed near-maximal tension. Both isoprenaline and sodium nitrite relaxed the K+-depolarized tissue though concentration-effect curves for both relaxants were moved to the right compared to those obtained in non-depolarized tissues. The maximal effect of sodium nitrite was markedly reduced. Intracellular electrophysiological recording showed that isoprenaline (0.01-1 mumol l-1) caused hyperpolarization and reduced or abolished slow wave discharge in trachealis muscle. These effects were accompanied by relaxation. Propranolol (1 mumol l-1) virtually abolished both the electrical and mechanical responses to isoprenaline (0.1 mumol l-1). Apamin (0.1 mumol l-1) did not alter the spontaneous electrical activity of trachealis cells or their electrical and mechanical responses to isoprenaline (0.1 mumol l-1). TEA (8 mmol l-1) caused depolarization and often increased slow wave amplitude and induced spike discharge. Isoprenaline (0.01 mumol l-1) failed to hyperpolarize TEA-treated trachealis cells. Higher concentrations of isoprenaline suppressed TEA-induced spasm, caused hyperpolarization and thereby increased slow wave or spike amplitude. Slow wave or spike frequency decreased as the hyperpolarization progressed but abolition of slow waves or spikes sometimes required more than 4 min exposure to isoprenaline. Procaine (5 mmol l-1) increased the amplitude of slow waves and induced spike discharge. Procaine markedly reduced the hyperpolarization induced by isoprenaline (0.1 and 1 mumol l-1) but had little effect on isoprenaline-induced relaxation. It is concluded that isoprenaline activates beta-adrenoceptors in guinea-pig trachealis and thereby evokes relaxation and hyperpolarization of the smooth muscle. The hyperpolarization does not involve the opening of apamin-sensitive K+-channels and it probably plays a supportive rather than a crucial role in the process by which isoprenaline-induced relaxation is achieved.

Action Potentials↗

PubMedDate not supplied

Renal developmental delay expressed by reduced glomerular number and its association with growth retardation in victims of sudden infant death syndrome and in "normal" infants.

In victims of sudden infant death syndrome (SIDS), renal development has been reported to be significantly impaired. In the present study, we used stereological techniques to estimate volume of kidney cortex and total number of glomeruli in a group of human infants. Infants were classified according to cause of death-SIDS or non-SIDS. Cases were further subdivided according to birth weight-normal birth weight (NBW) or low birth weight (LBW) (we were unable to identify any non-SIDS LBW infants for our study). No significant differences were found between NBW and LBW infants (irrespective of cause of death) for cortical volume, glomerular density, or total glomerular number (p > 0.140). Kidney cortical volume, glomerular density, and total glomerular number were not significantly different between SIDS and non-SIDS infants (p > 0.510). Glomerular number was only significantly less in SIDS infants of LBW (p = 0. 032) than in controls according to the Wilcoxon rank sum test; using the Kruskal-Wallis for one-way analysis, no significant difference was found (p > 0.010). These results contrast with those from previous studies, as a reduction in glomerular number was not noted in SIDS NBW infants, and the mean value for the control (non-SIDS NBW) group was significantly reduced (p < 0.01) from those of previous studies. This indicates that glomerular number reduction is seen in SIDS NBW and non-SIDS NBW cases and is therefore directly associated with growth retardation rather than with SIDS.

Birth Weight↗

PubMedDate not supplied

TRPC1 store-operated cationic channel subunit.

TRPC1 is a membrane protein that is highly conserved in mammals, amphibians and birds. It is widely expressed in cells throughout the body including in the heart and nervous system. Amino acid sequence analysis and over-expression studies indicate it is an ion channel that allows the transmembrane flux of small cations including sodium and calcium. In some cell types it is apparent that at least a fraction of TRPC1 exists in the plasma membrane. Inhibition of TRPC1 expression or block by TRPC1-specific antibody leads to attenuation of the plasma membrane calcium influx that occurs in response to depletion of calcium levels in sarcoplasmic or endoplasmic reticulum. TRPC1 would, therefore, seem to be a key subunit of store-operated channels (SOCs). TRPC1 is, nevertheless, unlikely to act alone. There is good evidence that it can heteromultimerise with the related proteins TRPC4, TRPC5 and polycystin-2; a tetrameric arrangement is envisaged, but not demonstrated. Like its relative in Drosophila, TRPC1 looks likely to function in a signalplex, a protein complex including inositol 1,4,5-triphosphate (IP(3)) receptor, plasma membrane calcium-ATPase, caveolin-1 and calmodulin. Its localisation in membranes is punctate and associated with functionally discrete calcium signals. TRPC1's function may not only be linked to SOCs but also to other cellular events including the nuclear translocation of the NFAT transcription factor. There is still much to be learned about this fundamental protein.

Amino Acid Sequence↗

PubMedDate not supplied

Activation thresholds of K(V), BK andCl(Ca) channels in smooth muscle cells in pial precapillary arterioles.

We have previously shown expression of voltage-gated K+ channels (K(V)) in smooth muscle of cerebral arterioles and suggested the channels function to oppose voltage-dependent Ca2+ entry. However, other studies indicate that large conductance Ca2+-activated K+ (BK) channels serve this function and chloride (Cl-) channels may have the opposite effect. In this study we compared the activation thresholds and absolute current amplitudes for K(V) channels, BK channels and Cl- channels at physiological membrane potentials in intact precapillary arterioles from the rabbit cerebral circulation. Patch-clamp recordings were made to measure current and membrane potential, and a video scan line was used to detect external diameter. Two strategies to determine the basal current-voltage relationship of BK channels showed the channels contributed current only at voltages positive of -35 mV, even though voltage-dependent Ca2+-entry occurred. Ca2+-activated and niflumic acid-sensitive Cl- current was detected but, between -50 and -10 mV, both BK and Cl- channel currents were much smaller and contributed less to the membrane potential compared with K(V) channel current. Furthermore, in the absence of an exogenous vasoconstrictor agent, block of K(V) channels but not BK or Cl- channels caused constriction, although in the presence of endothelin-1 block of BK or K(V) channels caused constriction. The data indicate K(V) channels are the first inhibitory mechanism to activate when there is depolarisation in precapillary arteriolar smooth muscle cells of the cerebral circulation.

4-Aminopyridine↗

PubMedDate not supplied

Association of hyaluronidase and breast adenocarcinoma invasiveness.

Hyaluronic acid (HA) is a component of the extracellular matrix (ECM) that exists as a high molecular weight polymer composed of alternating disaccharides, D-glucuronic acid and N-acetyl D-glucosamine. The interaction of hyaluronidase with HA results in the disruption of basement membrane integrity and produces an angiogenic response that has been implicated in tumor invasiveness and metastasis. Although hyaluronidase is present in several neoplasms, levels of hyaluronidase expression in breast cancer are not known. This investigation defines the correlation of elevated levels of hyaluronidase with breast adenocarcinoma invasiveness. Utilizing RT-PCR, RNA was extracted from paraffin embedded tissues (n=6) of patients diagnosed with fibrocystic breast changes, ductal carcinoma in situ (DCIS), and invasive adenocarcinoma. After constructing cDNA primers for base pairs 504 and 759 of the PH-20 gene (which is homologous to the hyaluronidase gene), PCR was performed and the products were visualized with ethidium bromide using gel electrophoresis. Invasive breast adenocarcinoma had a significantly higher level of hyaluronidase expression compared to other breast tissue samples; (32+/-15 vs. 8+/-3; p<0.03). Elevated levels of hyaluronidase correlated with invasive breast adenocarcinoma. Our data suggests that elevated levels of hyaluronidase are associated with breast adenocarcinoma invasive potential. Hyaluronidase may play an integral role in breast cancer invasion and metastasis.

Adenocarcinoma↗

PubMedDate not supplied

Increased hyaluronidase expression in more aggressive prostate adenocarcinoma.

Tumor metastasis involves a complex sequence of cellular and biochemical events of which tumor cell penetration through the basement membrane is an essential component. Disruption of basement membrane integrity by hyaluronidase has been implicated in the development of locally advanced and metastatic carcinoma. This investigation correlates hyaluronidase expression with pathologic prognostic variables for prostate adenocarcinoma. Paraffin samples (n=9) of patients receiving prostatectomies for clinical stage B adenocarcinoma were evaluated. RT-PCR was utilized to detect the presence of hyaluronidase. These results were correlated with the histological assessment of each sample. Gleason score was significantly higher in patients with RT-PCR detectable hyaluronidase (mean +/- SD 7.25+/-1.8 versus 4.17+/-1.0; p<0.05). Histological evidence of perineural invasion was seen in 83% of the specimens with detectable hyaluronidase, whereas none was found in hyaluronidase negative samples (p<0.05). A trend was seen between hyaluronidase and capsular invasion with 33% hyaluronidase positive tumor exhibiting evidence of capsular invasion, while no evidence in the hyaluronidase negative tumor. These data suggest that hyaluronidase may be involved in prostate adenocarcinoma progression and metastasis.

Biomarkers, Tumor↗

PubMedDate not supplied

Insulin-like growth factor-I receptor antagonism results in increased cytotoxicity of breast cancer cells to doxorubicin and taxol.

Therapeutic strategies for patients with advanced-stage adenocarcinoma of the breast frequently include the use of cytotoxic chemotherapy. Insulin-like growth factor I (IGF-I) receptor, a key factor in cell-cycle regulation, is frequently overexpressed in high-grade breast cancers. IGF-I receptor overexpression in these tumors may provide a target for novel molecular therapy against this disease. Early passage samples of estrogen-responsive (ER+) MCF 7 and estrogen receptor-negative (ER-) MDA-231 cells were cultured in semi-confluent conditions. Dose-titrations were performed for doxorubicin and taxol with receptor modulation using IGF-I or a competitive receptor inhibitor, alphaIR3. The addition of 100 ng/ml IGF-I resulted in a >2-fold mitogenic response in both ER+ and ER- cells. Receptor activation prior to the treatment with cytotoxic chemotherapeutic agents altered the pattern of response with a 26.3% increase in IC50. Doxorubicin and taxol both produced dose-related toxicity with IC50 of 0.05 microg/ml and 0.00 microg/ml respectively. The addition of alphaIR3 resulted in increased cytotoxicity in IGF-I stimulated cells compared with the use of doxorubicin or taxol alone. These results suggest that IGF-I receptor modulation alters the response to cytotoxic chemotherapeutic agents in breast cancer cells.

Antibodies, Monoclonal↗

PubMedDate not supplied

Nonfunctional parathyroid carcinoma.

Parathyroid carcinoma is a rare entity accounting for 0.5% to 5% of parathyroid neoplasia. Most of these malignancies present as functional hormone-producing masses with elevated serum levels of parathormone and calcium. These tumors may also be nonfunctional. Clinical detection of nonfunctioning parathyroid malignancies preoperatively is primarily based on symptoms of an expanding neck mass. This ominous complaint is typically accompanied with an advanced stage of the disease at initial diagnosis. Because there is a paucity of data in the literature regarding nonfunctioning parathyroid carcinoma, prognosis can not be readily assessed. In both functional and nonfunctional parathyroid carcinoma, early surgery has proven to be the only curative treatment approach whereas both chemotherapy and radiation therapy fail to produce systemic or regional benefit when used alone. Hence, parathyroid cancer should be considered in every patient evaluated for a neck mass regardless of the blood calcium and blood parathormone level.

Carcinoma↗