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1.
J Geophys Res Planets ; 128(1): e2022JE007480, 2023 Jan.
Article in English | MEDLINE | ID: mdl-37034458

ABSTRACT

The pressure sensors on Mars rover Perseverance measure the pressure field in the Jezero crater on regular hourly basis starting in sol 15 after landing. The present study extends up to sol 460 encompassing the range of solar longitudes from L s  âˆ¼ 13°-241° (Martian Year (MY) 36). The data show the changing daily pressure cycle, the sol-to-sol seasonal evolution of the mean pressure field driven by the CO2 sublimation and deposition cycle at the poles, the characterization of up to six components of the atmospheric tides and their relationship to dust content in the atmosphere. They also show the presence of wave disturbances with periods 2-5 sols, exploring their baroclinic nature, short period oscillations (mainly at night-time) in the range 8-24 min that we interpret as internal gravity waves, transient pressure drops with duration ∼1-150 s produced by vortices, and rapid turbulent fluctuations. We also analyze the effects on pressure measurements produced by a regional dust storm over Jezero at L s  âˆ¼ 155°.

2.
Trop Anim Health Prod ; 53(1): 62, 2021 Jan 03.
Article in English | MEDLINE | ID: mdl-33389254

ABSTRACT

Bovine rotavirus A (RVA) and bovine coronavirus (CoV) are the two main viral enteropathogens associated with neonatal calf diarrhea. The aim of the present work was to study the impact of group and individual housing systems in the epidemiology of RVA and CoV infection. Eleven calves reared in individual housing (FA) and nine calves in group housing (FB) were monitored during the first 7 weeks of life. Stool and serum samples were screened for RVA and CoV antigens by ELISA. IgG1 antibodies (Ab) to both antigens were also measured. From the 160 fecal samples collected, the proportion of positive samples to RVA and CoV was significantly higher in FB (23.6%) than in FA (9%) (p = 0.03). The geometric mean of colostral IgG1 Ab titers to CoV and RVA in FA (IgG1 anti-CoV 1024 and anti-RVA 1782.9) was lower than in FB (IgG1 anti-CoV 10,321.2 and anti-RVA 4096) at birth. Calves less than 2 weeks of life from FB had a higher risk of being infected by RVA (OR = 4.9; p = 0.01) and CoV (OR = 17.15; p = 0.01) than calves from FA. The obtained results showed that there was higher RVA and CoV shedding in group-housed calves than in individual-housed animals.


Subject(s)
Cattle Diseases/virology , Coronavirus Infections/veterinary , Housing, Animal , Rotavirus Infections/veterinary , Animals , Animals, Newborn , Argentina , Cattle , Cattle Diseases/epidemiology , Colostrum/immunology , Coronavirus Infections/virology , Coronavirus, Bovine , Dairying , Diarrhea/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Feces/virology , Female , Immunoglobulin G/immunology , Longitudinal Studies , Pregnancy , Rotavirus , Rotavirus Infections/virology , Virus Shedding
3.
Space Sci Rev ; 2162020 Dec 14.
Article in English | MEDLINE | ID: mdl-33536691

ABSTRACT

The Mars Regional Atmospheric Modeling System (MRAMS) and a nested simulation of the Mars Weather Research and Forecasting model (MarsWRF) are used to predict the local meteorological conditions at the Mars 2020 Perseverance rover landing site inside Jezero crater (Mars). These predictions are complemented with the COmplutense and MIchigan MArs Radiative Transfer model (COMIMART) and with the local Single Column Model (SCM) to further refine predictions of radiative forcing and the water cycle respectively. The primary objective is to facilitate interpretation of the meteorological measurements to be obtained by the Mars Environmental Dynamics Analyzer (MEDA) aboard the rover, but also to provide predictions of the meteorological phenomena and seasonal changes that might impact operations, from both a risk perspective and from the perspective of being better prepared to make certain measurements. A full diurnal cycle at four different seasons (Ls 0°, 90°, 180°, and 270°) is investigated. Air and ground temperatures, pressure, wind speed and direction, surface radiative fluxes and moisture data are modeled. The good agreement between observations and modeling in prior works [Pla-Garcia et al. in Icarus 280:103-113, 2016; Newman et al. in Icarus 291:203-231, 2017; Vicente-Retortillo et al. in Sci. Rep. 8(1):1-8, 2018; Savijarvi et al. in Icarus, 2020] provides confidence in utilizing these models results to predict the meteorological environment at Mars 2020 Perseverance rover landing site inside Jezero crater. The data returned by MEDA will determine the extent to which this confidence was justified.

4.
Astrobiology ; 19(7): 849-866, 2019 07.
Article in English | MEDLINE | ID: mdl-30964330

ABSTRACT

The shallow subsurface of Mars is extremely interesting as a possible microbial habitat because it becomes temporarily wet, it is shielded from radiation, and mixing by aeolian processes could provide the sources of energy and nutrients necessary for sustaining microbial life in it. The Modern Aqueous Habitat Reconnaissance Suite (MAHRS) was developed primarily to search for potentially habitable environments in the shallow subsurface of Mars and to study weathering, but it can also be used to search for potentially habitable environments in the shallow subsurface of other planetary bodies such as the Icy Worlds. MAHRS includes an instrument developed to measure regolith wetness and search for brine in the shallow subsurface of Mars, where it is most likely to be found. The detection of brine can aid in our understanding not only of habitability but also of geochemistry and aqueous weathering processes. Besides the regolith wetness sensor, MAHRS includes an electric field sensor, an optical microscope, and a radiometer developed to characterize the near-surface environment and study mixing by aeolian processes. MAHRS was designed to aid in the selection of optimum areas for sample collection for return to Earth.


Subject(s)
Ecosystem , Exobiology/instrumentation , Extraterrestrial Environment/chemistry , Mars , Anaerobiosis , Archaea/isolation & purification , Earth, Planet , Exobiology/methods , Geologic Sediments/chemistry , Geologic Sediments/microbiology , Ice Cover/chemistry , Ice Cover/microbiology , Oceans and Seas , Salts/analysis , Salts/chemistry , Water/analysis , Water/chemistry , Water Microbiology , Weather
5.
J Geophys Res Planets ; 124(11): 2780-2792, 2019 Nov.
Article in English | MEDLINE | ID: mdl-32025455

ABSTRACT

In situ measurements of relative humidity (RH) on Mars have only been performed by the Phoenix (PHX) and Mars Science Laboratory (MSL) missions. Here we present results of our recalibration of the PHX thermal and electrical conductivity probe (TECP) RH sensor. This recalibration was conducted using a TECP engineering model subjected to the full range of environmental conditions at the PHX landing site in the Michigan Mars Environmental Chamber. The experiments focused on the warmest and driest conditions (daytime) because they were not covered in the original calibration (Zent et al., 2010, https://doi.org/10.1029/2009JE003420) and previous recalibration (Zent et al., 2016, https://doi.org/10.1002/2015JE004933). In nighttime conditions, our results are in excellent agreement with the previous 2016 recalibration, while in daytime conditions, our results show larger water vapor pressure values. We obtain vapor pressure values in the range ~0.005-1.4 Pa, while Zent et al. (2016, https://doi.org/10.1002/2015JE004933) obtain values in the range ~0.004-0.4 Pa. Our higher daytime values are in better agreement with independent estimates from the ground by the PHX Surface Stereo Imager instrument and from orbit by Compact Reconnaissance Imaging Spectrometer for Mars. Our results imply larger day-to-night ratios of water vapor pressure at PHX compared to MSL, suggesting a stronger atmosphere-regolith interchange in the Martian arctic than at lower latitudes. Further, they indicate that brine formation at the PHX landing site via deliquescence can be achieved only temporarily between midnight and 6 a.m. on a few sols. The results from our recalibration are important because they shed light on the near-surface humidity environment on Mars.

6.
Sci Rep ; 8(1): 17576, 2018 12 04.
Article in English | MEDLINE | ID: mdl-30514860

ABSTRACT

In situ measurements by the Curiosity rover provide a unique opportunity for studying the effects of dust on assets placed at the surface of Mars. Here we use in situ measurements of solar UV radiation to quantify the seasonal and interannual variability of dust accumulation on the sensor on the rover deck. We show that the amount of dust accumulated on the sensor follows a seasonal cycle, with net dust removal during the perihelion season until Ls ~ 300°, and net dust deposition until the end of the aphelion season (Ls ~ 300°-180°). We use independent in situ measurements of atmospheric opacity and pressure perturbations in combination with numerical modeling, showing that daytime convective vortices and nighttime winds are likely responsible for the seasonal dust cleaning, with the role of nighttime wind being more important in Martian Year (MY) 32 than in MY 33 and that of daytime convective vortices being more important in MY 33 than in MY 32. The fact that the UV sensor is cleaner in MY 33 than in MY 32 indicates that natural cleaning events make solar energy an excellent candidate to power extended (multiannual) Mars missions at similar latitudes as the Curiosity rover.

7.
Phys Med Biol ; 63(19): 195009, 2018 09 28.
Article in English | MEDLINE | ID: mdl-30189420

ABSTRACT

Gas target systems have been used for decades on cyclotrons to produce radionuclides for medical imaging. However, the activity recovered from such targets is often lower than its theoretically predicted value. Past research has suggested that nuclide interactions with the walls of the target body may play a key role in the loss of recoverable radionuclide activity. Here, we consider gas targets and modify the standard radionuclide production equation by adding a loss term representing radionuclides depositing on the walls of the target. We derive the form of the deposition term based on a simple adsorption model which is then linearized by solving for leading order terms. The resulting production equation uses one fitting parameter to give an estimate of the recoverable activity produced in a target system, taking adsorption into account. The model is then fit to six data series, taken in-house and reported in the literature and a parity plot compares model predictions to experimental data. The model is able to better track the data than any previous models, and points towards a phenomenological understanding of adsorption in target systems.


Subject(s)
Gases/chemistry , Radiochemistry/methods , Radioisotopes/chemistry , Adsorption , Cyclotrons , Radiochemistry/instrumentation
8.
Rev. mex. ing. bioméd ; 39(1): 81-94, ene.-abr. 2018. tab, graf
Article in Spanish | LILACS-Express | LILACS | ID: biblio-902385

ABSTRACT

Resumen: En este artículo se presenta un enfoque para rehabilitación pasiva de miembro superior mediante la formulación de cuatro casos de estudio haciendo un análisis de las patologías y los ejercicios que se aplican. Para llevar a cabo la experimentación en los casos propuestos se registraron los datos de las trayectorias de las articulaciones del brazo de un paciente realizando los ejercicios de rehabilitación con un terapeuta. Se diseñó el exoesqueleto ERMIS de siete grados de libertad para emular los movimientos anatómicos en el brazo durante la rehabilitación a partir de los requerimientos de los casos de estudio. Para validar el funcionamiento del exoesqueleto en los casos se simuló el modelo dinámico del ERMIS y se compararon los datos con los datos muestreados de los ejercicios. Al final se presentan los resultados obtenidos de los ejercicios realizados con el exoesqueleto, obteniendo en la precisión un desempeño promedio del 95% en los movimientos de hombro, codo y muñeca al emular la terapia con timón.


Abstract: This paper presents an approach for passive upper limb rehabilitation based on four case studies by analyzing the pathologies and exercises that are applied. To carry out the experimentation in the proposed cases, the data from the trajectories of the patient's arm articulations were registered, performing the rehabilitation exercises with a therapist. The ERMIS exoskeleton´s seven degrees of freedom was designed to emulate the anatomical movements in the arm during rehabilitation from the requirements of the case studies. To validate the exoskeleton performance in the study cases, the ERMIS's dynamic model was simulated and the data were compared with the sampled data of the exercises. At the end, the results obtained from exoskeleton exercises emulating rudder therapy, where shoulder, elbow and wrist movements were showed with an accuracy of 95%.

9.
Trop Anim Health Prod ; 50(4): 801-806, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29274052

ABSTRACT

The aim of this work was to study the epidemiology and the harmful effect of gastrointestinal nematodes (GINs) on beef cattle in the piedmont forest and grassland region of northwestern Argentina, between March 2015 and March 2016. Sixty weaned female calves were divided into three groups: STG-calves treated systematically with 200 mcg/kg moxidectin every 40-50 days; LTG-calves treated as usually locally implemented, with 200 mcg/kg ivermectin on May 4 and August 5; and UTG-calves not receiving treatment. Moreover, a group of 20 untreated first-calving cows was monitored. Eggs per gram of faeces (epg) and faeces cultures were performed. Live weight gain (LWG) was recorded. Differences among groups were compared using analysis of variance and Tukey test. At the start of the experiment, mean epg was from moderate to high (595 ± 480); therefore, this group was treated in May. Mean UTG epg peaked (907 ± 754) in mid-winter, decreasing toward spring. LTG epg only decreased partially after treatment, showing 54.2% of ivermectin efficacy. Epg values of STG were always negligible values (95.8% of moxidectin efficacy). The dominant nematode genus was Cooperia (81.9%), followed by Haemonchus (15.5%). The effect of treatments was evident from middle autumn, with STG showing a significantly (p < 0.002) higher LWG than the other groups. Toward the end of the trial, LWG of STG and LTG increased by 27.2 and 8.2%, respectively, to those of UTG. This preliminary study showed a winter epg peak and a negative effect of GINs on LWG, even in moderately infected calves.


Subject(s)
Antinematodal Agents/therapeutic use , Cattle Diseases/epidemiology , Ivermectin/therapeutic use , Macrolides/therapeutic use , Rhabditida Infections/veterinary , Animals , Antinematodal Agents/pharmacology , Argentina/epidemiology , Cattle , Cattle Diseases/parasitology , Cattle Diseases/prevention & control , Drug Evaluation, Preclinical , Feces , Female , Gastrointestinal Diseases/parasitology , Gastrointestinal Diseases/prevention & control , Gastrointestinal Diseases/veterinary , Gastrointestinal Tract/parasitology , Haemonchus/isolation & purification , Ivermectin/pharmacology , Macrolides/pharmacology , Nematoda , Ovum , Parasite Egg Count/statistics & numerical data , Parasite Egg Count/veterinary , Rhabditida Infections/prevention & control , Weight Gain/drug effects
11.
Astrobiology ; 16(12): 937-948, 2016 12.
Article in English | MEDLINE | ID: mdl-27912028

ABSTRACT

In the last few years, water ice and salts capable of melting this ice and producing liquid saline water (brine) have been detected on Mars. Moreover, indirect evidence for brine has been found in multiple areas of the planet. Here, we simulate full diurnal cycles of temperature and atmospheric water vapor content at the Phoenix landing site for the first time and show experimentally that, in spite of the low Mars-like chamber temperature, brine forms minutes after the ground temperature exceeds the eutectic temperature of salts in contact with water ice. Moreover, we show that the brine stays liquid for most of the diurnal cycle when enough water ice is available to compensate for evaporation. This is predicted to occur seasonally in areas of the polar region where the temperature exceeds the eutectic value and frost or snow is deposited on saline soils, or where water ice and salts coexist in the shallow subsurface. This is important because the existence of liquid water is a key requirement for habitability. Key Words: Mars-Ice-Perchlorates-Brine-Water-Raman spectroscopy. Astrobiology 16, 937-948.


Subject(s)
Circadian Rhythm , Extraterrestrial Environment , Mars , Salts/analysis , Ice/analysis , Image Processing, Computer-Assisted , Perchlorates/analysis , Spectrum Analysis, Raman , Temperature , Vibration , Water/chemistry
12.
J Geophys Res Planets ; 119(8): 1822-1838, 2014 Aug.
Article in English | MEDLINE | ID: mdl-26213666

ABSTRACT

The analysis of the surface energy budget (SEB) yields insights into soil-atmosphere interactions and local climates, while the analysis of the thermal inertia (I) of shallow subsurfaces provides context for evaluating geological features. Mars orbital data have been used to determine thermal inertias at horizontal scales of ∼104 m2 to ∼107 m2. Here we use measurements of ground temperature and atmospheric variables by Curiosity to calculate thermal inertias at Gale Crater at horizontal scales of ∼102 m2. We analyze three sols representing distinct environmental conditions and soil properties, sol 82 at Rocknest (RCK), sol 112 at Point Lake (PL), and sol 139 at Yellowknife Bay (YKB). Our results indicate that the largest thermal inertia I = 452 J m-2 K-1 s-1/2 (SI units used throughout this article) is found at YKB followed by PL with I = 306 and RCK with I = 295. These values are consistent with the expected thermal inertias for the types of terrain imaged by Mastcam and with previous satellite estimations at Gale Crater. We also calculate the SEB using data from measurements by Curiosity's Rover Environmental Monitoring Station and dust opacity values derived from measurements by Mastcam. The knowledge of the SEB and thermal inertia has the potential to enhance our understanding of the climate, the geology, and the habitability of Mars.

13.
Rev. chil. cir ; 62(3): 262-267, jun. 2010. ilus
Article in Spanish | LILACS | ID: lil-562726

ABSTRACT

Single photon emission computed tomography and computed tomography (SPECT/CT), integrates a gamma camera and a CT scan and is effective for the location of parathyroid adenomas. We report a 55 years old male and a 80 years old female with primary hyperparathyroidism. In both cases the 99mTc-Sestamibi parathyroid scintigraphy detected a functioning nodule whose presence was confirmed with SPECT/CT.


El SPECT/CT integra una gamacámara y un escáner radiológico en un solo equipo híbrido que fusiona la imagen de la cintigrafía SPECT (tomografía computada de fotón único) con la imagen morfológica obtenida con un escáner de baja intensidad, sin movilizar al paciente, en una perfecta correspondencia anátomo-funcional, permitiendo identificar con exactitud la localización de un adenoma paratiroideo. Se describen dos pacientes con hiperparatiroidismo primario, con ecotomografía convencional negativa, donde el SPECT/CT demostró la localización exacta de adenomas de ubicación no habitual.


Subject(s)
Humans , Male , Female , Middle Aged , Aged, 80 and over , Adenoma , Parathyroid Neoplasms , Tomography, Emission-Computed, Single-Photon , Adenoma/surgery , Adenoma/pathology , Hyperparathyroidism, Primary , Parathyroid Neoplasms/surgery , Parathyroid Neoplasms/pathology , Preoperative Care
14.
Rev Chilena Infectol ; 27(1): 60-4, 2010 Feb.
Article in Spanish | MEDLINE | ID: mdl-20140317

ABSTRACT

Sexually transmitted diseases are highly prevalent and a public health concern worldwide. Herpes simplex virus (HSV) and human papilloma virus (HPV) are described. The development of rapid, sensible and specific diagnostic assays has been difficult because of their pathogenic features. In the last years, molecular biology based techniques allowed a better and wider range of diagnosis, as in the HPV-cancer association. In this article, standardized diagnostic methodologies for HSV and HPV are reviewed.


Subject(s)
Herpes Simplex/diagnosis , Papillomaviridae , Papillomavirus Infections/diagnosis , Sexually Transmitted Diseases/diagnosis , Sexually Transmitted Diseases/virology , Simplexvirus , Female , Herpes Simplex/transmission , Humans , Male , Papillomaviridae/genetics , Papillomaviridae/immunology , Papillomavirus Infections/transmission , Simplexvirus/genetics , Simplexvirus/immunology
15.
Rev. chil. infectol ; 27(1): 60-64, feb. 2010.
Article in Spanish | LILACS | ID: lil-537173

ABSTRACT

Sexually transmitted diseases are highly prevalent and a public health concern worldwide. Herpes simplex virus (HSV) and human papilloma virus (HPV) are described. The development of rapid, sensible and specific diagnostic assays has been difficult because of their pathogenic features. In the last years, molecular biology based techniques allowed a better and wider range of diagnosis, as in the HPV-cancer association. In this article, standardized diagnostic methodologies for HSV and HPV are reviewed.


Las Infecciones virales de transmisión sexual son altamente prevalentes y constituyen un problema de salud pública en el mundo. Entre los agentes que se contagian por esta vía, se describe acerca de virus herpes simplex (HSV) y virus papiloma humano (HPV). Las características patogénicas de estas infecciones han dificultado la implementación de técnicas diagnósticas rápidas, sensibles y específicas para el diagnóstico clínico habitual. En los últimos años las metodologías diagnósticas sustentadas en la biología molecular han permitido mej orar y ampliar el rango de diagnóstico posible para estos agentes infecciosos y relacionarlos con otras patologías, como es el caso de HPV y cáncer. En el presente artículo se revisan metodologías diagnósticas implementadas para el diagnóstico microbiológico de HSV y de HPV.


Subject(s)
Female , Humans , Male , Herpes Simplex/diagnosis , Papillomaviridae , Papillomavirus Infections/diagnosis , Simplexvirus , Sexually Transmitted Diseases/diagnosis , Sexually Transmitted Diseases/virology , Herpes Simplex/transmission , Papillomaviridae/genetics , Papillomaviridae/immunology , Papillomavirus Infections/transmission , Simplexvirus/genetics , Simplexvirus/immunology
16.
Rev Chilena Infectol ; 26(4): 356-9, 2009 Aug.
Article in Spanish | MEDLINE | ID: mdl-19802405

ABSTRACT

Eczema herpeticum is an extensive vesicular eruption caused by cutaneous dissemination of herpes simplex virus in patients who have underlying skin conditions or epidermal barrier disruption. Herpes simplex virus type 1 (HSV 1) is the most common etiology. This is the first report in Chile of a case of eczema herpeticum caused by HSV 2 in a patient with Darier's disease, We review the literature and emphasize the use of sensitive and specific assays as real time polymerase chain reaction, that allow to initiate timely the correct treatment.


Subject(s)
Herpesvirus 2, Human , Kaposi Varicelliform Eruption/virology , Acyclovir/therapeutic use , Cloxacillin/therapeutic use , Humans , Kaposi Varicelliform Eruption/drug therapy , Male , Middle Aged , Polymerase Chain Reaction
17.
Rev. chil. infectol ; 26(4): 356-359, ago. 2009. ilus
Article in Spanish | LILACS | ID: lil-527880

ABSTRACT

Eczema herpeticum is an extensive vesicular eruption caused by cutaneous dissemination of herpes simplex virus in patients who have underlying skin conditions or epidermal barrier disruption. Herpes simplex virus type 1 (HSV 1) is the most common etiology. This is the first report in Chile of a case of eczema herpeticum caused by HSV 2 in a patient with Darier' s disease, We review the literature and emphasize the use of sensitive and specific assays as real time polimerase chain reaction, that allow to initiate timely the correct treatment.


El eccema herpético es una erupción vesicular extensa causada por la diseminación cutánea del virus herpes simplex, en pacientes con trastornos cutáneos previos o daño de la barrera epidérmica. La etiología más frecuente es virus herpes simplex tipo 1 (VHS 1), aunque se han reportado casos de virus herpes simplex tipo 2 (VHS 2). Presentamos el primer reporte en Chile de eccema herpético por VHS 2 en un paciente con enfermedad de Darier. Realizamos una revisión de la literatura médica y destacamos la importancia del uso de técnicas sensibles y específicas como la reacción de polimerasa en cadena (RPC) en tiempo real, que posibilitan el inicio precoz de la terapéutica correcta.


Subject(s)
Humans , Male , Middle Aged , Kaposi Varicelliform Eruption/virology , Acyclovir/therapeutic use , Cloxacillin/therapeutic use , Kaposi Varicelliform Eruption/drug therapy , Polymerase Chain Reaction
18.
Am J Physiol Heart Circ Physiol ; 291(3): H1147-57, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16679513

ABSTRACT

Microvascular endothelial cells involved in angiogenesis are exposed to an acidic environment that is not conducive for growth and survival. These cells must exhibit a dynamic intracellular (cytosolic) pH (pHcyt) regulatory mechanism to cope with acidosis, in addition to the ubiquitous Na+/H+ exchanger and HCO3--based H+-transporting systems. We hypothesize that the presence of plasmalemmal vacuolar-type proton ATPases (pmV-ATPases) allows microvascular endothelial cells to better cope with this acidic environment and that pmV-ATPases are required for cell migration. This study indicates that microvascular endothelial cells, which are more migratory than macrovascular endothelial cells, express pmV-ATPases. Spectral imaging microscopy indicates a more alkaline pHcyt at the leading than at the lagging edge of microvascular endothelial cells. Treatment of microvascular endothelial cells with V-ATPase inhibitors decreases the proton fluxes via pmV-ATPases and cell migration. These data suggest that pmV-ATPases are essential for pHcyt regulation and cell migration in microvascular endothelial cells.


Subject(s)
Cell Membrane/enzymology , Cell Movement/physiology , Endothelium, Vascular/physiology , Vacuolar Proton-Translocating ATPases/metabolism , Animals , Cell Membrane/physiology , Cells, Cultured , Endothelium, Vascular/cytology , Endothelium, Vascular/enzymology , Hydrogen-Ion Concentration , Immunohistochemistry , Microcirculation/physiology , Rats , Rats, Inbred BB , Sodium-Hydrogen Exchangers/physiology
19.
Biochem Pharmacol ; 57(9): 1037-46, 1999 May 01.
Article in English | MEDLINE | ID: mdl-10796074

ABSTRACT

A major obstacle for the effective treatment of cancer is the phenomenon of multidrug resistance (MDR) exhibited by many tumor cells. Many, but not all, MDR cells exhibit membrane-associated P-glycoprotein (P-gp), a drug efflux pump. However, most mechanisms of MDR are complex, employing P-gp in combination with other, ill-defined activities. Altered cytosolic pH (pHi) has been implicated to play a role in drug resistance. In the current study, we investigated mechanisms of pHi regulation in drug-sensitive (MCF-7/S) and drug-resistant human breast cancer cells. Of the drug-resistant lines, one contained P-gp (MCF-7/DOX; also referred to as MCF-7/D40) and one did not (MCF-7/MITOX). The resting steady-state pHi was similar in the three cell lines. In addition, in all the cell lines, HCO3- slightly acidified pHi and increased the rates of pHi recovery after an acid load, indicating the presence of anion exchanger (AE) activity. These data indicate that neither Na+/H+ exchange nor AE is differentially expressed in these cell lines. The presence of plasma membrane vacuolar-type H+-ATPase (pmV-ATPase) activity in these cell lines was then investigated. In the absence of Na+ and HCO3-, MCF-7/S cells did not recover from acid loads, whereas MCF-7/MITOX and MCF-7/DOX cells did. Furthermore, recovery of pHi was inhibited by bafilomycin A1 and NBD-Cl, potent V-ATPase inhibitors. Attempts to localize V-ATPase immunocytochemically at the plasma membranes of these cells were unsuccessful, indicating that V-ATPase is not statically resident at the plasma membrane. Consistent with this was the observation that release of endosomally trapped dextran was more rapid in the drug-resistant, compared with the drug-sensitive cells. Furthermore, the drug-resistant cells entrapped doxorubicin into intracellular vesicles whereas the drug-sensitive cells did not. Hence, it is hypothesized that the measured pmV-ATPase activity in the drug-resistant cells is a consequence of rapid endomembrane turnover. The potential impact of this behavior on drug resistance is examined in a companion manuscript.


Subject(s)
Antineoplastic Agents/pharmacology , Breast Neoplasms/enzymology , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Proton-Translocating ATPases/metabolism , Vacuolar Proton-Translocating ATPases , Benzopyrans , Bicarbonates/metabolism , Biological Transport , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Cell Compartmentation , Doxorubicin/pharmacology , Drug Resistance, Multiple/genetics , Endosomes/physiology , Female , Fluorescent Dyes/metabolism , Gene Expression Regulation, Neoplastic/drug effects , Humans , Hydrogen-Ion Concentration , Naphthols/metabolism , Proton-Translocating ATPases/biosynthesis , Proton-Translocating ATPases/genetics , Rhodamines/metabolism , Sodium-Hydrogen Exchangers , Tumor Cells, Cultured , Vacuoles/enzymology
20.
Cell Physiol Biochem ; 8(3): 158-74, 1998.
Article in English | MEDLINE | ID: mdl-9617478

ABSTRACT

Based on studies using high-affinity Ca2+ probes (dissociation constant (Kd) = 0.15-0.3 microM), steady-state [Ca2+]in is believed to be in the nanomolar range in most cells. However, probes with lower affinity indicate that [Ca2+]in may increase to micromolar levels during activation of specific cell functions, e.g., contraction. These conclusions rely on accurate knowledge of the Kd of the dyes for Ca2+. Mag-Fura-2 (also known as Furaptra) is a low-affinity Ca2+ indicator (Kd ca. 50 microM) which has been used for such studies. In the present work, Mag-Fura-2 is shown to respond to changes in cytosolic Ca2+ in the submicromolar range. In vitro, and in situ titration of Mag-Fura-2 in A7r5 cells, demonstrate that Mag-Fura-2 exhibits both high- and low-affinity for Ca2+. Moreover, pH affects both high and low affinity Ca2+ binding site. Since Mag-Fura-2 has been used to study Ca2+ within specific subcellular compartments, the present observations indicate that knowledge of factors such as ambient pH of these compartments is required to accurately interpret Ca2+ responses. Furthermore, the sensitivity of Mag-Fura-2 at submicromolar levels must be considered for accurate determination of Ca2+ in specific compartments believed to exhibit high micromolar levels of Ca2+.


Subject(s)
Calcium/metabolism , Fluorescent Dyes/metabolism , Fura-2/analogs & derivatives , Binding Sites , Cell Line , Fura-2/metabolism , Hydrogen-Ion Concentration , Muscle, Smooth, Vascular/cytology , Muscle, Smooth, Vascular/metabolism , Sensitivity and Specificity , Spectrometry, Fluorescence
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