Instrument Calibration of the Interface Region Imaging Spectrograph (IRIS) Mission.

The Interface Region Imaging Spectrograph (IRIS) is a NASA small explorer mission that provides high-resolution spectra and images of the Sun in the 133 - 141 nm and 278 - 283 nm wavelength bands. The IRIS data are archived in calibrated form and made available to the public within seven days of observing. The calibrations applied to the data include dark correction, scattered light and background correction, flat fielding, geometric distortion correction, and wavelength calibration. In addition, the IRIS team has calibrated the IRIS absolute throughput as a function of wavelength and has been tracking throughput changes over the course of the mission. As a resource for the IRIS data user, this article describes the details of these calibrations as they have evolved over the first few years of the mission. References to online documentation provide access to additional information and future updates.

Evidence of nonthermal particles in coronal loops heated impulsively by nanoflares.

The physical processes causing energy exchange between the Sun's hot corona and its cool lower atmosphere remain poorly understood. The chromosphere and transition region (TR) form an interface region between the surface and the corona that is highly sensitive to the coronal heating mechanism. High-resolution observations with the Interface Region Imaging Spectrograph (IRIS) reveal rapid variability (~20 to 60 seconds) of intensity and velocity on small spatial scales (≲500 kilometers) at the footpoints of hot and dynamic coronal loops. The observations are consistent with numerical simulations of heating by beams of nonthermal electrons, which are generated in small impulsive (≲30 seconds) heating events called "coronal nanoflares." The accelerated electrons deposit a sizable fraction of their energy (≲10(25) erg) in the chromosphere and TR. Our analysis provides tight constraints on the properties of such electron beams and new diagnostics for their presence in the nonflaring corona.

Hot explosions in the cool atmosphere of the Sun.

The solar atmosphere was traditionally represented with a simple one-dimensional model. Over the past few decades, this paradigm shifted for the chromosphere and corona that constitute the outer atmosphere, which is now considered a dynamic structured envelope. Recent observations by the Interface Region Imaging Spectrograph (IRIS) reveal that it is difficult to determine what is up and down, even in the cool 6000-kelvin photosphere just above the solar surface: This region hosts pockets of hot plasma transiently heated to almost 100,000 kelvin. The energy to heat and accelerate the plasma requires a considerable fraction of the energy from flares, the largest solar disruptions. These IRIS observations not only confirm that the photosphere is more complex than conventionally thought, but also provide insight into the energy conversion in the process of magnetic reconnection.

Prevalence of small-scale jets from the networks of the solar transition region and chromosphere.

As the interface between the Sun's photosphere and corona, the chromosphere and transition region play a key role in the formation and acceleration of the solar wind. Observations from the Interface Region Imaging Spectrograph reveal the prevalence of intermittent small-scale jets with speeds of 80 to 250 kilometers per second from the narrow bright network lanes of this interface region. These jets have lifetimes of 20 to 80 seconds and widths of ≤300 kilometers. They originate from small-scale bright regions, often preceded by footpoint brightenings and accompanied by transverse waves with amplitudes of ~20 kilometers per second. Many jets reach temperatures of at least ~10(5) kelvin and constitute an important element of the transition region structures. They are likely an intermittent but persistent source of mass and energy for the solar wind.

The unresolved fine structure resolved: IRIS observations of the solar transition region.

The heating of the outer solar atmospheric layers, i.e., the transition region and corona, to high temperatures is a long-standing problem in solar (and stellar) physics. Solutions have been hampered by an incomplete understanding of the magnetically controlled structure of these regions. The high spatial and temporal resolution observations with the Interface Region Imaging Spectrograph (IRIS) at the solar limb reveal a plethora of short, low-lying loops or loop segments at transition-region temperatures that vary rapidly, on the time scales of minutes. We argue that the existence of these loops solves a long-standing observational mystery. At the same time, based on comparison with numerical models, this detection sheds light on a critical piece of the coronal heating puzzle.

On the prevalence of small-scale twist in the solar chromosphere and transition region.

The solar chromosphere and transition region (TR) form an interface between the Sun's surface and its hot outer atmosphere. There, most of the nonthermal energy that powers the solar atmosphere is transformed into heat, although the detailed mechanism remains elusive. High-resolution (0.33-arc second) observations with NASA's Interface Region Imaging Spectrograph (IRIS) reveal a chromosphere and TR that are replete with twist or torsional motions on sub-arc second scales, occurring in active regions, quiet Sun regions, and coronal holes alike. We coordinated observations with the Swedish 1-meter Solar Telescope (SST) to quantify these twisting motions and their association with rapid heating to at least TR temperatures. This view of the interface region provides insight into what heats the low solar atmosphere.

The origins of hot plasma in the solar corona.

The Sun's outer atmosphere, or corona, is heated to millions of degrees, considerably hotter than its surface or photosphere. Explanations for this enigma typically invoke the deposition in the corona of nonthermal energy generated by magnetoconvection. However, the coronal heating mechanism remains unknown. We used observations from the Solar Dynamics Observatory and the Hinode solar physics mission to reveal a ubiquitous coronal mass supply in which chromospheric plasma in fountainlike jets or spicules is accelerated upward into the corona, with much of the plasma heated to temperatures between ~0.02 and 0.1 million kelvin (MK) and a small but sufficient fraction to temperatures above 1 MK. These observations provide constraints on the coronal heating mechanism(s) and highlight the importance of the interface region between photosphere and corona.

In vitro priming of human lymphocytes. I. IL-2 and IL-4 requirements.

Induction of antigen-dependent IgM and IgG responses by human in vitro spleen cell cultures were supported by addition of defined lymphokines. Whereas exposure to IL-2 alone throughout both immunization and cultivation period supported the highest antigen-directed responses, antibodies of higher relative affinity to the immunizing antigen were secreted under conditions of shorter IL-2 exposure. Continuous presence of IL-2 supported the antigen-driven responses for up to 15 days, the later part of which was characterized by a very low relative affinity index value. IL-2 supported cultures produced up to 55 times and 36 times more IgM and IgG, respectively, than cultures without added IL-2. Addition of IL-2-neutralizing antibodies throughout the cultivation period abrogated all responses. Addition of IL-4 alone had negligible effect on the antigen-directed IgM responses but supported antigen-independent IgG responses. Neutralization of IL-4 alone had negligible effect on the IgM and IgG responses, whereas neutralization of IL-4 during IL-2 support, resulted in reduction of antigen dependency of responses. Delayed IL-4 neutralization (24 hours) restored some of the antigen sensitivity. IL-4 reduced IL-2-induced, antigen-directed immunoglobulin responses but supported increased antigen-induced, antigen-directed responses, resulting in maximal antigen-specific responses. IL-4 reduced the IL-2-induced immunoglobulin production. IL-2 supported cell division, whereas IL-4 supported prolonged survival. Flow cytometry tests showed that IL-2 primarily induced CD8+ cells (T suppressor/cytotoxic) and OKT-10+ cells (plasma cells) cells, whereas the number of B1+ cells (B cells) decreased. IL-4 induced slight increases in the amount of B1+ cells and CD4+ cells (T helper).

In vitro priming of human lymphocytes. II. Induction of antigen-specific IgG responses by repeated antigen stimulation.

A protocol by which a human spleen cell culture could be subjected to both a primary and a secondary type immunization in vitro has been developed. Primary immunized cultures required the addition of autologous fresh cells at the time of antigen re-exposure for generation of optimal antigen-specific IgG responses. Antigen re-exposure (secondary stimulation) was done 10 days after initial immunization. Subsequent responses were characterized by novel or greatly enhanced antigen-directed IgG responses compared to primary antigen stimulation. Re-exposure of antigen to 10-day old human spleen cell cultures, without addition of fresh cells, did not result in measurable responses. Lethal irradiation of the fresh cell component of a restimulation culture showed that the IgG responses were derived from the original culture and not the fresh cells. The presence of antigen during both the primary and the secondary immunization was a prerequisite for induction of antigen-directed IgG responses, as spleen cell preparations that had been cultivated without antigen, during the primary stimulation period or the secondary stimulation period or both, did not show antigen-dependent IgG responses after the secondary cultivation. Induction of IgG responses by restimulation was supported by IL-2, but only when IL-2 was added to both the primary and the secondary culture according to a strict protocol. IL-4 added during the second antigen exposure supported antigen-directed responses, whereas IL-4 added during the initial antigen exposure abrogated all antigen-dependent responses.

Production of antigen-specific human monoclonal antibodies from in vitro-primed human splenocytes.

We have developed culture conditions for human lymphocytes that support primary in vitro immune responses to protein Ag of either human or nonhuman origin. We now show that these primed B cells can be efficiently immortalized by fusion with a heterohybrid fusion partner to generate human, Ag-specific IgM or IgG antibody-producing heterohybridomas at a rate of 17 to 50 hybrids/10(6) lymphocytes fused. Approximately 50% of the Ig-secreting clones were stable with respect to Ig secretion. Levels of secretion attained with terminal cultures ranged from less than 1 to 100 micrograms/ml. Fusions of cells between 2 and 5 days after initiation of in vitro exposure to Ag produced more Ag-reactive and Ag-specific antibodies than fusions at 1 day or fusions performed after 5 days. Ag-reactive hybrids could be isolated at frequencies of 3 to 10%, depending on antigenicity of the immunogen. Foreign proteins, horse spleen ferritin, and a murine monoclonal Ig, induced higher percentages of Ag-reactive mAb than immunization with the human-derived ferritin. Ag-reactive IgG mAb were produced at relatively high frequency, depending on immunization conditions and the nature of the Ag. The strategy for identification of the best hybrids included early elimination of unstable hybridomas and of hybridomas producing broadly cross-reactive antibody, followed by evaluation of units of Ag reactivity/micrograms Ig. Ferritin-specific mAb selected according to these criteria showed immunocytochemical reactivity with ferritin-containing tissues and apparent affinities in the range of 10(7) to 10(8)/mol.

Effects of 5-azacytidine, sodium butyrate, and phorbol esters on amino acid transport system A in a kidney epithelial cell line, MDCK: evidence for multiple mechanisms of regulation.

Neutral amino acid transport by system A was investigated in the epithelial cell lines MDCK and MDCK-T1. The latter line is a chemically induced, oncogenically transformed line derived from MDCK. Inducers of differentiation, sodium butyrate and 5-azacytidine, and a tumor promoter, TPA, were used as probes to delineate pathways of regulation involved in system A response to a variety of physiological conditions and agents. Azacytidine, an inhibitor of DNA methylation, and butyrate, an enhancer of histone acetylation, inhibited expression of system A, had little effect on system ASC, and slightly stimulated system L. Inhibition of system A expression by butyrate and azacytidine occurred under different conditions. Increases in system A activity due to amino acid starvation or transformation were inhibited by butyrate but not by azacytidine. Repressed system A activity, normally observed in the presence of high levels of amino acids, was more sensitive to azacytidine than to butyrate. The tumor promoter, TPA, stimulated system A activity in MDCK cells under normal growth conditions but did not stimulate activity in amino acid-starved MDCK cells or in MDCK-T1 cells. Stimulation of system A activity by TPA was prevented by prior exposure to butyrate but not to azacytidine. These results suggest 1) that system A expression observed in growing amino-acid-repressed MDCK cells is modulated by an azacytidine-sensitive mechanism and 2) that the elevated expression of system A activity induced by amino acid starvation, by chemical transformation to MDCK-T1, and by TPA is modulated by a butyrate-sensitive mechanism.

Polarity of neutral amino acid transport and characterization of a broad specificity transport activity in a kidney epithelial cell line, MDCK.

The Madin-Darby canine kidney (MDCK) cell line was investigated with respect to the cellular polarity of amino acid transport in early confluent versus late confluent cultures. Early confluent cultures could take up amino acids from the apical and the basolateral sides of the cell layer via amino acid transport Systems A, ASC, and L. However, in late confluent cultures the activities of Systems A and L were clearly localized to the basolateral surface of the cell monolayer. In addition to the presence of systems A, ASC, and L, a novel activity, measurable under conditions used for quantitating System ASC, was found to be active in the apical membrane of these cells. This transporter, termed System G (for general), recognized basic and neutral amino acids with high affinity and acidic amino acids with lower affinity. System G exhibited broad substrate specificity, strict cation specificity, and a broad pH optimum with maximal activity at acidic pH. The activity of System G was relatively low after growth in serum-containing medium but was induced in a defined medium. Induction of System G activity was dependent upon the presence of prostaglandin E1. The broad substrate specificity, low pH optimum, and Na+ dependence suggest that System G may function in apical membranes as an energy-dependent transport route during reabsorption of amino acids from the kidney tubule lumen.

Sodium entry pathways in renal epithelial cell lines.

Na+ entry into kidney epithelial cells occurs by a multiplicity of pathways. Established cell lines such as the A6 cells, derived from the collecting duct of the kidney of Xenopus laevis, MDCK cells, from the distal tubule of a dog kidney, and the LLC-PK1 cells, originating from the proximal tubule of a pig kidney, provide excellent model cell systems for the detailed characterization and isolation of the proteins which comprise these entry pathways. Major pathways of Na+ entry include the amiloride-sensitive Na+ channel, the amiloride-sensitive Na+/H+ antiporter, and the loop diuretic-sensitive NaCl/KCl symporter. While the former two systems have been shown to exhibit an apical location in epithelial cells so far examined, the last system may be localized to either the basolateral or apical surface, depending on the transport function of the cell. Nutrient/Na+ symporters such as the glucose, phosphate, and p-aminohippurate symporters may all be localized to the apical surfaces of proximal tubular cells, but other systems, including those specific for neutral amino acids, may predominate in the basolateral surface or be distributed between the two membranes. Studies concerned with the catalytic, structural, and regulatory properties of these transport systems serve not only to characterize the individual translocators in established cell lines, but also to suggest their physiological functions in intact kidney tissues.

Methionine-sensitive glycolysis in transformed cells.

Glycolysis in several tumor cell lines grown in tissue culture was inhibited by methionine. Kirsten murine sarcoma virus-transformed rat kidney cells (K-NRK) were inhibited 60-75% by 10 mM methionine, whereas normal rat kidney (NRK-49F) cells showed little or no inhibition. Inhibition of glycolysis in K-NRK cells was manifest 2-4 hr after exposure to the amino acid. Glycolysis in a chemically transformed cell line of Madin-Darby canine kidney cells was also sensitive to methionine, but maximal inhibition (75%) required 18-24 hr of incubation with the amino acid. Under the same conditions glycolysis in the nontransformed canine cells was less than 20% inhibited by methionine. In Ehrlich ascites tumor cells grown in tissue culture, 10 mM methionine inhibited glycolysis by about 50%. Inhibition of glycolysis, even by 50 mM methionine, was rapidly reversible. Within 2 hr after removal of methionine the rate of glycolytic activity was restored to that observed in control cells. Furthermore, inhibition by methionine required a minimum level (7%) of serum in the growth medium and inhibition was not sensitive to cycloheximide. Only amino acids that are transported by system A (including the nonmetabolized analogue methylaminoisobutyric acid) specifically inhibited glycolysis in tumor cells. The only exception was phenylalanine, which was toxic to both transformed and normal cell lines.

Stimulation of glycolysis and amino acid uptake in NRK-49F cells by transforming growth factor beta and epidermal growth factor.

Glycolysis in normal resting rat kidney cells (NRK-49F) was stimulated by a 2-hr exposure to transforming growth factors prior to assay. Transforming growth factor beta (TGF-beta) was effective when added alone, and further addition of epidermal growth factor (EGF) had little effect. The stimulation by TGF-beta was abolished when cycloheximide was present during the incubation, suggesting that protein synthesis is required for the effect. Incubation of the cells with 25 mM methionine abolished the stimulation of glycolysis by TGF-beta. The uptake of methylaminoisobutyrate via system A was stimulated by either TGF-beta or EGF. The greater than 3-fold stimulation of uptake by 1 ng of pure TGF-beta per ml was usually somewhat enhanced on addition of 0.5 ng of EGF per ml. Moreover, an antiserum against EGF receptor partially depressed the response to TGF-beta, suggesting some overlapping interactions of EGF and TGF-beta.

Adaptive regulatory control of System A transport activity in a kidney epithelial cell line (MDCK) and in a transformed variant (MDCK-T1).

Adaptive regulatory control of System A activity was investigated using MDCK cells and a chemically induced, oncogenic transformant of MDCK cells, MDCK-T1. Within 7 hours after transfer to an amino-acid-deficient medium, A activity of subconfluent MDCK cells had maximally derepressed, but this activity in confluent MDCK cells and in subconfluent transformed cells showed little capacity for derepression. Amino-acid-starved, subconfluent MDCK cells were used to study trans-inhibition and repression of A activity by individual amino acids. Trans-inhibition and repression were defined as the cycloheximide-insensitive and cycloheximide-sensitive components, respectively, of the total inhibition. Trans-inhibition correlated well with substrate affinity, but repression did not. Trans-inhibition and repression were further characterized using alpha-(methylamino) isobutyric acid (mAIB), a trans-inhibitor, and glutamate, an effective repressor. The apparent initial T 1/2 for inhibition by mAIB in the presence of cycloheximide was 0.5 hours, while that for repression by glutamate was 4.7 hours. Half-maximal inhibition by mAIB and repression by glutamate occurred at approximately 0.02 mM and 0.07 mM, respectively. Reversal of trans-inhibition by methionine occurred in the presence of cycloheximide within 1-4 hours after removal of methionine. The A system of the transformed MDCK-T1 cells showed elevated activity, little capacity for derepression, resistance to repression by amino acids, but retention of sensitivity to trans-inhibition. Kinetic analysis of mAIB uptake indicated that the A system of MDCK-T1 cells has become kinetically more complex in a manner which resembled amino-acid-starved rather than amino-acid-fed MDCK cells. These results suggest that the A system of MDCK-T1 cells has become resistant to adaptive regulatory control.

Hormonal regulation of the System A amino acid transport adaptive response mechanism in a kidney epithelial cell line (MDCK).

When mammalian cells are starved for amino acids, the activity of the A amino acid transport system increases, a phenomenon called adaptive regulation. We have examined the effects of those factors which support Madin-Darby canine kidney (MDCK) cell growth in a defined medium on the derepression of System A activity. Of the five factors which supported MDCK cell growth, insulin was found to be an absolute requirement for derepression. In contrast, PGE1 was a negative controlling factor for the transport system. Growth of MDCK cells in the absence of PGE1 resulted in elevated System A activity which derepressed poorly upon amino acid starvation. Kinetic analysis of alpha-(methylamino) isobutyric acid (mAIB) uptake as a function of substrate concentration showed that the elevated A activity observed when cells were grown in the absence of PGE1 was kinetically similar to the activity induced by starvation for amino acids. Transport of mAIB by amino-acid-fed cells grown in the presence of PGE1 was characterized by a linear Eadie-Hofstee graph and by a relatively low Vmax. Transport by cells starved for amino acids or by cells grown in the absence of PGE1 was characterized by biphasic kinetics for mAIB transport and by elevated Vmax values. An influence of growth factors on the inactivation of derepressed A activity was also observed. In the presence of cycloheximide the rate of loss of A activity in amino-acid-starved cells was 1/4-1/2 that of amino-acid-fed cells. Insulin slowed inactivation in the absence of most amino acids in a protein-synthesis-independent manner, but insulin did not influence the more rapid inactivation observed in amino-acid-fed cells. These results indicate that the level of System A activity observed in response to regulation by amino acids represents a balance between carrier synthesis and inactivation, which can be positively or negatively influenced by growth factors.

Characterization of chemically and virally transformed variants of Madin-Darby canine kidney (MDCK) epithelial cells.

Oncogenic derivatives of Madin-Darby canine kidney (MDCK) cells were isolated in the nude mouse, and nononcogenic anchorage-independent transformants were isolated in vitro following chemical mutagenesis in vitro. These transformed cell lines as well as a Moloney sarcoma virus (MSV) transformed line were characterized with respect to their serum and anchorage requirements, growth rates, final saturation densities, and sensitivities to contact inhibition. None of these in vitro growth characteristics were found to correlate with tumorigenicity in nude mice. One tumorigenic clone, MDCK-T1, was characterized with respect to serum-free growth requirements, cAMP production, and ornithine decarboxylase (ODC) activity. These cells exhibited a significant reduction in the PGE1 requirement for growth, they produced higher levels of cAMP, and they expressed a reduced level of ODC activity relative to the parental MDCK cells. These findings may reflect changes in growth control mechanisms which accompany kidney epithelial cell tumorigenesis and suggest that the study of transformed lines derived in this manner could lead to the identification of in vitro properties which are associated with malignancy.