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PubMed1976

Evaluation of the H+/site ratio of mitochondrial electron transport from rate measurements.

The mitochondrial H+/site ratio (i.e. the number of protons ejected per pair of electrons traversing each of the energy-conserving sites of the respiratory chain) has been evaluated employing a new experimental approach. In this method the rates of oxygen uptake and H+ ejection were measured simultaneously during the initial period of respiration evoked by addition of succinate to aerobic, rotenone-inhibited, de-energized mitochondria. Either K+, in the presence of valinomycin, or Ca2+, was used as mobile cation to dissipate the membrane potential and allow quantitative H+ ejection into the medium. The H+/site ratio observed with this method in the absence of precautions to inhibit the uptake of phosphate was close to 2.0, in agreement with values obtained using the oxygen pulse technique (Mitchell, P. and Moyle, J. (1967) Biochem. J. 105, 1147-1162). However, when phosphate movements were eliminated either by inhibition of the phosphate-hydroxide antiporter with N-ethylamaleimide or by depleting the mitochondria of their endogenous phosphate content, H+/site ratios close to 4.0 were consistently observed. This ratio was independent of the concentration of succinate, of mitochondrial protein, of pH between 6 and 8, and of ionic composition of the medium, provided that sufficient K+ (plus valinomycin) or Ca2+ were present. Specific inhibitors of the hydrolysis of endogenous ATP or transport of other ions (adenine nucleotides, tricarboxylates, HCO3-, etc.) were shown not to affect the observed H+/site ratio. Furthermore, the replacement of succinate by alpha-glycerol phosphate, a substrate which is oxidized on the outer surface of the inner membrane and thus does not need to enter the matrix, gave the same H+/site ratios as did succinate. It is concluded that the H+/site ratio of mitochondrial electron transport, when phosphate movements are eliminated, may be close to 4.0.

Animals↗
PubMed2004

Mobile guidelines for pharmacologic treatment of hypertension.

Mobile health technologies provide case studies with potentially rather huge impacts. Examples are provided e.g. by guidelines involving pharmacologic treatment. Knowledge and reasoning need to interact with information management, and often involves utility of various devices. Well organised databases for pharmacological information are also necessary for successful engineering of mobile extensions in these case studies.

Antihypertensive Agents↗
PubMed1977

[Stimulation of the reflex response of the anticoagulant system by intermediate product-1 of prothrombin proteolysis].

Injection of the intermediate 1 of prothrombin proteolysis into the frog's systemic circulation mobilized the anticoagulating and the fibrinolytic potential of the organism. This is not so in frogs with destroyed c.n. s. Electrical activity of the carotid chemoreceptors in considerably higher after perfusion with intermediate 1 of the frog's humorally isolated carotid labyrinth, as well as the total coagulation time, the total fibrinolytic activity, and the non-fermentative fibrinolysis in the systematic circulation. The data obtained suggest that the reflex activation of the physiological anticoagulating system can be induced can be induced by intermediate 1 which has the structure similar to thrombin but no clotting activity.

Animals↗
PubMed1987

[In vitro demonstration of histamine biosynthesis from carnosine by kidneys of pregnant mice].

Kidneys of pregnant mice synthesize histamine when incubated in the presence of carnosine, manganese, and pyridoxal phosphate. Intensity of biosynthesis increases linearly with the amount of enzyme and the incubation time. The reaction can only be catalysed by two enzymes that are located in kidneys and act in succession: carnosinase, which hydrolyzes carnosine into its two moieties, and histidine decarboxylase, which transforms histidine, a product of carnosine degradation, into histamine. The biosynthesis of histamine from carnosine seems to increase with the progress of pregnancy. In nonpregnant mice, kidneys do not effect this biosynthesis. The above results directly demonstrate that carnosine may be used for histamine synthesis when the activity of histidine decarboxylase is high, as in pregnant mouse kidney. Vertebrate carnosine, its role still enigmatic, might thus be mainly a potential histidine reservoir that would be mobilized any time there is a significant requirement for histidine, such as for histamine biosynthesis.

Animals↗
PubMed1994

Roles of v-erbA homodimers and heterodimers in mediating dominant negative activity by v-erbA.

v-erbA, a viral oncogenic homolog of thyroid hormone receptor (TR), blocks the effect of T3 in TR-mediated transcription. The mechanism(s) for this dominant negative effect by v-erbA on TRs is unknown but may involve competition between v-erbA and TR-containing complexes for binding to thyroid hormone response elements (TREs) and/or protein-protein interactions between v-erbA and TR. To investigate these potential mechanisms, we used the electrophoretic mobility shift assay to compare in vitro translated v-erbA and TR alpha binding to two TREs-chick lysozyme TRE (F2) and direct repeat TRE (DR4). v-erbA bound as a homodimer to these TREs, whereas TR alpha bound as a homodimer and monomer. T3 decreased TR alpha homodimer binding to the TREs as we reported previously; however, surprisingly, high concentrations of T3 (10(-6) M) also decreased v-erbA homodimer binding to the TREs. Additionally, v-erbA formed heterodimers with nuclear proteins such as retinoid X receptor and T3 receptor auxiliary protein as well as with TR alpha. These dimers remained bound to DNA in the presence of T3. Finally, v-erbA could not mediate ligand-dependent transcriptional activation even at 10(-6) M T3 but could block ligand-dependent TR-mediated transactivation in co-transfection experiments. v-erbA also exhibited differential dominant negative activity on F2 and DR4 suggesting that half-site sequence and/or orientation may influence v-erbA-dominant negative activity. In sum, there are multiple v-erbA complexes that bind to TREs in the presence of T3, which all may contribute to v-erbA's dominant negative effect on TR-mediated transcription by competing with TR-containing complexes for binding to TREs.

Base Sequence↗
PubMed1993

[Ballistic (Lithoclast) and hydro-electric (Riwolith) endo-ureteral lithotripsy. Report of 60 cases].

25 hydroelectric endoureteric lithotripsies (Group 1, Riwolith) and 35 ballistic endoureteric lithotripsies (Group 2, Lithoclast) were analysed. 60 stones were treated, including 18 pelvic stones (Group 1 : 10, Group 2 : 8), 39 iliolumbar stones (Group 1 : 14, Group 2 : 25), and 3 stones of the ureteropelvic junction (Group 1 : 1, Group 2 : 2). The mean maximal diameter was 9.9 mm (Group 1 : 8.2, Group 2 : 11). Complementary Dormia extraction was performed in 24 cases (Group 1 : 8, Group 2 : 16), 55 double J stents were implanted (Group 1 : 24, Group 2 : 31) and 23 immediate ECL sessions were performed (Group 1 : 8, Group 2 : 15). Overall, 57 good results were recorded, but 3 complete failures of fragmentation and 2 severe ureteric lesions were attributed to hydroelectric lithotripsy. The Lithoclast, much less traumatic, had a greater stone fragmentation potential, but its efficacy on very mobile stones is limited due to its more intense stone propulsion effect.

Adult↗
PubMed2005

Local-scale density-dependent survival of mobile organisms in continuous habitats: an experimental test using Atlantic salmon.

For organisms with restricted mobility, density dependence may occur on spatial scales much smaller than that of the whole population. Averaging densities over whole populations in such organisms gives a more or less inaccurate description of the real variation in competitive intensity over time and space. The potential for local density dependence in more mobile organisms is less well understood, particularly for organisms living in continuous habitats. To test for local density-dependent processes in such an organism, we manipulated egg density (the number of eggs nest(-1)) among ten artificial nests of Atlantic salmon along an 1,848-m long river during two consecutive years. Eggs in different nests were given unique thermal otolith-banding patterns to allow identification of juvenile nest origin. At capture, 1-2 months after emergence, the spatial distribution of juveniles reflected nest locations, with the median absolute dispersal distance being 92 and 41 m in the 2 years. Estimated nest-specific survival rates were strongly negatively related to hatched-egg density in both years (r(2)=0.72 and 0.62), despite dramatic differences in overall mean survival (0.22 and 0.02). Thus, density-dependent survival following emergence in Atlantic salmon juveniles occurs on spatial scales much smaller than that of whole populations. The consistency across years suggests that the phenomenon is likely to occur over most environmental conditions. Our observation of local-scale density dependence is consistent with strong juvenile territoriality, which forces individuals emerging in high-initial density areas to disperse farther, and a high cost (metabolic or predation) of dispersal. We conclude that for mobile organisms with patchy distributions of propagules and constrained juvenile dispersal, increased emphasis on local-scale dynamics should enable a more mechanistic understanding of population regulation even in continuous habitats, and hence increase the predictive power of population models.

Analysis of Variance↗
PubMed2006

Residual mobility of instrumented and non-fused segments in thoracolumbar spine fractures.

The surgical management of thoracolumbar fractures presents potential benefits. However, the surgery solve the instability by fusion of mobile segments. We incorporate in our treatment algorithms, the use of restricted arthrodesis at injured levels, regardless of longer instrumentations, as well as the use of non-fused transitory stabilizations, based on the conviction that in non-fused segments without traumatic disc injury, mobility persists once the instrumentation is removed. The goals of this study were to compare the mobility of non-fused segments after hardware removal to a normal range of motion and to find prognostic pre-op imaging patterns. We reviewed 21 consecutive patients who underwent surgery with preservation of mobile segments (non-fused segments included in the construction) in order to recover mobility after removal of instrumentation, performed between 1995 and 2001. All patients were treated by indirect reduction with posterior transpedicular instrumentation. Clinical and radiological outcome was analyzed after an average follow-up of 46.6 months. Satisfactory subjective outcome results were obtained in 94.7%. The dynamic radiological follow-up study showed 75% (21 segments) with normal or decreased range of motion (ROM) and 25% (7 segments) without mobility. The non-fused segments with hardware removal before 10 months of evolution presented a normal or decreased mobility in 83.2% while the segments with hardware removal after 10 months showed 68.8% of mobility. The intervertebral disc (IVD)'s with normal initial MRI morphology preserved their mobility in 81.9%. Complications occurred in four patients: two superficial wound infections and two patients presented a late fracture of one USS Schanz. The results of this study prove that in thoracolumbar fractures, non-fused spinal segments included in pedicular instrumentation maintained mobility in a high percentage once the hardware is removed. 75% of the segments presented a normal or decreased ROM.

Adult↗
PubMed1968

Surface charge of giant axons of squid and lobster.

A method is described for the determination of the electrophoretic mobility of single, isolated, intact, giant axons of squid and lobster. In normal physiological solutions, the surface of hydrodynamic shear of these axons is negatively charged. The lower limit of the estimated surface charge density is -1.9 x 10(-8) coul cm(-2) for squid axons, -4.2 x 10(-8) coul cm(-2) for lobster axons. The electrophoretic mobility of squid axons decreases greatly when the applied transaxial electric field is made sufficiently intense; action potential propagation is blocked irreversibly by transaxial electric fields of the same intensity. The squid axon recovers its mobility hours later and is then less affected by transaxial fields. Eventually, a state is reached in which the transaxial field irreversibly reverses the sign of the surface charge. In contrast, there is no obvious effect of electric field on the mobility of lobster axons. The mobility of lobster axons becomes undetectable in the presence of Th(4+) at a concentration which blocks the action potential, and in the presence of La(3+) at a concentration which does not affect propagation. Quinine does not alter lobster axon mobility at a concentration which blocks action potential conduction. Replacement of extracellular Na(+) by K(+) is without effect upon lobster axon mobility. The electrophysiological implications of the results are discussed.

Action Potentials↗
PubMed2000

The probability of quantal secretion within an array of calcium channels of an active zone.

A Monte Carlo analysis has been made of calcium dynamics in submembranous domains of active zones in which the calcium contributed by the opening of many channels is pooled. The kinetics of calcium ions in these domains has been determined using simulations for channels arranged in different geometries, according to the active zone under consideration: rectangular grids for varicosities and boutons and lines for motor-nerve terminals. The effects of endogenous fixed and mobile buffers on the two-dimensional distribution of free calcium ions at these active zones are then given, together with the extent to which these are perturbed and can be detected with different affinity calcium indicators when the calcium channels open stochastically under an action potential. A Monte Carlo analysis of how the dynamics of calcium ions in the submembranous domains determines the probability of exocytosis from docked vesicles is also presented. The spatial distribution of exocytosis from rectangular arrays of secretory units is such that exocytosis is largely excluded from the edges of the array, due to the effects of endogenous buffers. There is a steeper than linear increase in quantal release with an increase in the number of secretory units in the array, indicating that there is not just a local interaction between secretory units. Conditioning action potentials promote an increase in quantal release by a subsequent action potential primarily by depleting the fixed and mobile buffers in the center of the array. In the case of two parallel lines of secretory units exocytosis is random, and diffusion, together with the endogenous calcium buffers, ensures that the secretory units only interact over relatively short distances. As a consequence of this and in contrast to the case of the rectangular array, there is a linear relationship between the extent of quantal secretion from these zones and their length, for lengths greater than a critical value. This Monte Carlo analysis successfully predicts the relationship between the size and geometry of active zones and the probability of quantal secretion at these, the existence of quantal versus multiquantal release at different active zones, and the origins of the F1 phase of facilitation in synapses possessing different active zone geometries.

Action Potentials↗
PubMed2002

Brain-derived neurotrophic factor induces long-lasting Ca2+-activated K+ currents in rat visual cortex neurons.

Brain-derived neurotrophic factor (BDNF) increases postsynaptic intracellular Ca2+ and modulates synaptic transmission in various types of neurons. Ca2+-activated K+ currents, opened mainly by intracellular Ca2+ elevation, contribute to hyperpolarization following action potentials and modulate synaptic transmission. We asked whether BDNF induces Ca2+-activated K+ currents by postsynaptic elevation of intracellular Ca2+ in acutely dissociated visual cortex neurons of rats. Currents were analysed using the nystatin-perforated patch clamp technique and imaging of intracellular Ca2+ mobilization with fura-2. At a holding potential of -50 mV, BDNF application (20 ng/mL) for 1-2 min induced an outward current (IBDNF-OUT; 80.0 +/- 29.0 pA) lasting for more than 90 min without attenuation in every neuron tested. K252a (200 nm), an inhibitor of Trk receptor tyrosine kinase, and U73122 (3 microm), a specific phospholipase C (PLC)-gamma inhibitor, suppressed IBDNF-OUT completely. IBDNF-OUT was both charybdotoxin- (600 nm) and apamin- (300 nm) sensitive, suggesting that this current was carried by Ca2+-activated K+ channels. BAPTA-AM (150 microm) gradually suppressed IBDNF-OUT. Fura-2 imaging revealed that a brief application of BDNF elicited a long-lasting elevation of intracellular Ca2+. These results show that BDNF induces long-lasting Ca2+-activated K+ currents by sustained intracellular Ca2+ elevation in rat visual cortex neurons. While BDNF, likely acting through the Trk B receptor, was necessary for the induction of long-lasting Ca2+-activated K+ currents via intracellular Ca2+ elevation, BDNF was not necessary for the maintenance of this current.

Action Potentials↗
PubMed2002

Nup98 is a mobile nucleoporin with transcription-dependent dynamics.

Nucleoporin 98 (Nup98), a glycine-leucine-phenylalanine-glycine (GLFG) amino acid repeat-containing nucleoporin, plays a critical part in nuclear trafficking. Injection of antibodies to Nup98 into the nucleus blocks the export of most RNAs. Nup98 contains binding sites for several transport factors; however, the mechanism by which this nucleoporin functions has remained unclear. Multiple subcellular localizations have been suggested for Nup98. Here we show that Nup98 is indeed found both at the nuclear pore complex and within the nucleus. Inside the nucleus, Nup98 associates with a novel nuclear structure that we term the GLFG body because the GLFG domain of Nup98 is required for targeting to this structure. Photobleaching of green fluorescent protein-Nup98 in living cells reveals that Nup98 is mobile and moves between these different localizations. The rate of recovery after photobleaching indicates that Nup98 interacts with other, less mobile, components in the nucleoplasm. Strikingly, given the previous link to nuclear export, the mobility of Nup98 within the nucleus and at the pore is dependent on ongoing transcription by RNA polymerases I and II. These data give rise to a model in which Nup98 aids in direction of RNAs to the nuclear pore and provide the first potential mechanism for the role of a mobile nucleoporin.

Animals↗
PubMed1998

Detection of high mobility group I HMGI(Y) protein in the diagnosis of thyroid tumors: HMGI(Y) expression represents a potential diagnostic indicator of carcinoma.

Hyperplastic or neoplastic proliferative lesions of thyroid follicular epithelium consist of a spectrum, ranging from nodular hyperplasia to undifferentiated (anaplastic) carcinoma, and usually present as palpable thyroid nodules. Thyroid nodules are a common occurrence in the general population, but only a small proportion of them are eventually diagnosed as carcinoma. The difficulty in objectively identifying those thyroid nodules that are malignant to avoid unnecessary surgery, combined with the range and effectiveness of the available therapeutic options in those patients who do, indeed, have thyroid carcinoma, has prompted the search for tumor markers and prognostic indicators. The high mobility group I (HMGI) proteins represent a class of nuclear proteins involved in the regulation of chromatin structure and function. HMGI(Y), one of the members of this class, is expressed at high levels during embryogenesis and in malignant tumors but at generally low levels in normal adult human tissues. Previous work on a limited number of thyroid samples suggested that the detection of the HMGI(Y) proteins may provide a clinically useful diagnostic tool. To verify this assumption, we analyzed HMGI(Y) expression by a combination of immunohistochemistry and reverse transcription-PCR in 358 thyroid tissue samples that were representative of the spectrum of thyroid tumor pathology. HMGI(Y) was detectable in 18 of 19 follicular carcinomas, 92 of 96 papillary carcinomas, and 11 of 11 undifferentiated (anaplastic) carcinomas but in only 1 of 20 hyperplastic nodules, 44 of 200 follicular adenomas, and 0 of 12 normal tissue samples. The correlation between HMGI(Y) expression and a diagnosis of carcinoma was highly significant (P < 0.0001). We also prospectively collected and analyzed for HMGI(Y) expression by immunohistochemistry and reverse transcription-PCR in 12 fine needle aspiration biopsies from 10 patients who subsequently underwent surgical removal of a solitary thyroid nodule. HMGI(Y) was detectable only in the four fine needle aspiration biopsies, corresponding to the thyroid nodules that were definitively diagnosed as carcinomas after surgery (two follicular carcinomas and two papillary carcinomas). The remaining eight samples (six follicular adenomas and two samples consisting of normal follicular cells) were negative. The findings of this study confirm the differential expression of HMGI(Y) in thyroid neoplasia and indicate the HMGI(Y) protein as a potential marker for thyroid carcinoma.

Adenocarcinoma, Follicular↗
PubMed2003

Mass spectrometry-compatible ICH (International Conference on Harmonization) impurity analysis with a high-pH mobile phase advantages and pitfalls.

Recent advances in bonding chemistry and novel silica synthesis have significantly extended the pH range of silica-based HPLC columns. This extended range now enables the analysis of water-soluble basic drugs at high pH without ion-pairing reagents, thus offering an alternative approach to assay or impurity analyses. This paper describes the many advantages and potential pitfalls of using high-pH mobile phases in the development of MS-friendly LC gradient impurity analytical methods for water-soluble basic drugs under International Conference on Harmonization (ICH) guidelines. Operating at high-pH provides excellent peak shapes and retention, and accentuates selectivity differences of structurally similar impurities and degradants. However, several problems unique to the use of high-pH mobile phases, such as column lifetime, robustness of pH adjustments, peak fronting, and on-column dimerization, were encountered. Each of these problems is discussed with its respective remedy.

Hydrogen-Ion Concentration↗
PubMed2002

Changes in integrin expression are associated with altered homing properties of Lin(-/lo)Thy1.1(lo)Sca-1(+)c-kit(+) hematopoietic stem cells following mobilization by cyclophosphamide/granulocyte colony-stimulating factor.

OBJECTIVE: Although migration of hematopoietic stem cells (HSC) is essential for normal hematopoiesis and successful hematopoietic cell transplantation, little is known about the mechanisms that underlie this movement. We have sought to characterize the factors that regulate HSC migration by analyzing changes in expression of particular adhesion receptors associated with cyclophosphamide/granulocyte colony-stimulating factor (Cy/G-CSF)-induced HSC mobilization. METHODS: Expression by Lineage(-/lo)Thy1.1(lo)Sca-1(+)c-kit(+) HSC of members of the beta1 integrin family of adhesion molecules was assessed in untreated or Cy/G-CSF-treated mice by multiparameter flow cytometry. In parallel, the in vivo homing properties of normal and mobilized HSC were compared following intravenous transfer of fluorescently marked HSC. RESULTS: Normal adult HSC express high levels of several beta1 integrin family members. Following Cy/G treatment, bone marrow HSC selectively downregulate alpha 2 integrin expression and upregulate alpha 5 expression. HSC found in the blood following Cy/G-CSF treatment express significantly lower levels of multiple integrins than their bone marrow and/or splenic counterparts. Changes in integrin expression by blood-borne HSC correlate with a 50% decrease in their ability to home to the bone marrow in short-term assays, and with previously observed defects in competitive engraftment by these HSC. Similar reductions in bone marrow (BM) homing are observed for BM HSC treated with alpha 4 integrin function blocking mAb prior to injection. Modulation of integrin expression induced by mobilization was not associated with cell-cycle progression. CONCLUSION: Changes in integrin expression and function are associated with HSC mobilization and likely significantly affect the engraftment potential of hematopoietic stem cells.

Animals↗
PubMed1996

Radical thoracoscopic esophagectomy for cancer.

BACKGROUND: Much of the morbidity of conventional esophagectomy for cancer is thought to relate to the thoracotomy wound and while transhiatal esophagectomy removes the need for a thoracotomy, it is not oncologically sound. Videothoracoscopy could potentially provide an oncologically sound means for resecting the thoracic esophagus without the need for a thorcotomy. METHODS: Between June 1991 and June 1994, thoracoscopic mobilization of the thoracic esophagus combined with radical lymphadenectomy was attempted in 24 patients as part of three-stage esophagectomy for cancer (5 squamous and 19 adenocarcinomas). Mean age was 59 years (range 43-76). Eight patients were ASA grade I, 10 were ASA II, and 6 ASA III. Two patients had early lesions (T1N0) but all other cancers were T2 or T3. Dissection of the thoracic esophagus was attempted via a right-sided approach, followed by a laparotomy and a cervical incision. RESULTS: The thoracoscopic procedure was successful in 22 patients; it was abandoned in one patient with dense pleural adhesions and in another with inoperable tumor. Mean duration of the thoracic component was 184 min(120-330). There were three post-operative deaths. Ten further patients had major complications. Median post-operative stay was 18 days(9-129). Mean node harvest was 13 nodes(6-28). Two-year survival (cancer specific) was 33%. CONCLUSIONS: Radical thoracoscopic mobilization of the esophagus is feasible, but the potential for complications remains high and requires further study.

Adenocarcinoma↗