Biomedical subjects

H W Kim

Publications and source records attributed to H W Kim.

At least 19 recordsLinked to original sources

PubMed1992

Regulation of the skeletal sarcoplasmic reticulum Ca(2+)-ATPase by phospholamban and negatively charged phospholipids in reconstituted phospholipid vesicles.

The Ca(2+)-ATPase of skeletal sarcoplasmic reticulum was purified and reconstituted in proteoliposomes containing phosphatidylcholine (PC). When reconstitution occurred in the presence of PC and the acidic phospholipids, phosphatidylserine (PS) or phosphatidylinositol phosphate (PIP), the Ca(2+)-uptake and Ca(2+)-ATPase activities were significantly increased (2-3 fold). The highest activation was obtained at a 50:50 molar ratio of PS:PC and at a 10:90 molar ratio of PIP:PC. The skeletal SR Ca(2+)-ATPase, reconstituted into either PC or PC:PS proteoliposomes, was also found to be regulated by exogenous phospholamban (PLB), which is a regulatory protein specific for cardiac, slow-twitch skeletal, and smooth muscles. Inclusion of PLB into the proteoliposomes was associated with significant inhibition of the initial rates of Ca(2+)-uptake, while phosphorylation of PLB by the catalytic subunit of cAMP-dependent protein kinase reversed the inhibitory effects. The effects of PLB on the reconstituted Ca(2+)-ATPase were similar in either PC or PC:PS proteoliposomes, indicating that inclusion of negatively charged phospholipid may not affect the interaction of PLB with the skeletal SR Ca(2+)-ATPase. Regulation of the Ca(2+)-ATPase appeared to involve binding with the hydrophilic portion of phospholamban, as evidenced by crosslinking experiments, using a synthetic peptide which corresponded to amino acids 1-25 of phospholamban. These findings suggest that the fast-twitch isoform of the SR Ca(2+)-ATPase may be also regulated by phospholamban although this regulator is not expressed in fast-twitch skeletal muscles.

Affinity Labels↗

PubMed1992

CD28-stimulated IL-2 gene expression in Jurkat T cells occurs in part transcriptionally and is cyclosporine-A sensitive.

CD28 is a glycoprotein expressed as a homodimer on the surface of a major subset of human T cells. Previous studies have shown that proliferation of peripheral blood T cells involving the CD28 pathway is associated with cyclosporine A (CsA) resistant IL-2 gene expression. This pathway was shown to specifically regulate the stability of mRNA for several lymphokines including IL-2. We have investigated the expression of the IL-2 gene in the Jurkat cell line, J32 clone, induced by CD28 stimulation. Cross-linked anti-CD28 mAb alone was sufficient to induce the release of small amounts of IL-2 (256 U/ml). The CD28-mediated IL-2 release was enhanced with simultaneous engagement of CD28 and CD2 or CD28 and CD3 molecules. Hybrid constructs in which the human IL-2 gene 5' flanking region drives luciferase expression (pIL-2-Luc) were used to help delineate whether the CD28 pathway activates the IL-2 gene transcriptionally. Costimulation of cells with CD28 mAb and either PHA or CD2 mAb induced a 20- to 90-fold increase in the expression of pIL-2-Luc as well as IL-2 release. Costimulation with CD28 mAb plus PMA gave only five- to sevenfold increase in enhancer activity. In contrast, no enhancer activity was detected after stimulation with CD28 or CD2 mAb alone. Both IL-2 release and pIL-2-Luc activity were inhibited by CsA in J32 cells. The degree of CsA inhibition was concentration dependent and was similar in cells stimulated with either CD28 mAb or CD3 mAb. Maximum inhibition was achieved with 1 microgram/ml of CsA. Studies with internal deletion mutations of the IL-2 gene 5' flanking sequence revealed that as with stimulation through the TCR pathway, the CD28 pathway requires 5' flanking sequences located within 500 bp of the transcription start site. These results are the first direct evidence that the triggering of CD28 molecule is sufficient to induce IL-2 release in J32 cells. Furthermore these studies strongly indicate that IL-2 gene expression induced by CD28 stimulation occurs, in part, transcriptionally and is CsA sensitive in these cells.

Antigens, CD↗

PubMed1992

Mouse phospholamban gene expression during development in vivo and in vitro.

To establish a murine model that may allow for definition of the precise role of phospholamban in myocardial contractility through selective perturbations in the phospholamban gene, we initiated studies on the role of phospholamban in the murine heart. Intact beating hearts were perfused in the absence or presence of isoproterenol, and quantitative measurements of cardiac performance were obtained. Isoproterenol stimulation was associated with increases in the affinity of the sarcoplasmic reticulum Ca2+ pump for Ca2+ that were due to phospholamban phosphorylation. To assess the regulation of phospholamban gene expression during murine development, Northern blot and polymerase chain reaction analyses were used. Phospholamban mRNA was first detected in murine embryos on the ninth day of development (the time when the cardiac tube begins to contract). In murine embryoid bodies, which have been shown to recapitulate several aspects of cardiogenesis, phospholamban mRNA was detected on the seventh day (the time when spontaneous contractions are first observed). Only those embryoid bodies that exhibited contractions expressed phospholamban transcripts, and these were accompanied by expression of the protein, as revealed by immunofluorescence microscopy. Sequence analysis of the cDNA encoding phospholamban in embryoid bodies indicated complete homology to that in adult hearts. The deduced amino acid sequence of murine phospholamban was identical to rabbit cardiac phospholamban but different from dog cardiac and human cardiac phospholamban by one amino acid. These data suggest that phospholamban, the regulator of the Ca(2+)-ATPase in cardiac sarcoplasmic reticulum, is present very early in murine cardiogenesis in utero and in vitro, and this may constitute an important determinant for proper development of myocardial contractility.

Amino Acid Sequence↗

PubMed1992

Evaluating new red cell substitutes: a critical analysis of toxicity models.

Because red cell substitutes (RCS) will improve oxygen delivery to tissues rendered hypoxic or anoxic they are presumed to be effective. Prior to clinical application the safety--toxicity of any proposed solution must be established. Models used to demonstrate efficacy do not necessarily evaluate toxicity. Infusion of human hemoglobin (Hb) solutions into animals raises one issue of immune response but obviates looking at the human response to similar materials. Differentiating the cause of an immunologic effect may be difficult. Effects seen in acute models may not have chronic implications; they may be transients of no consequence. Differentiating the effects of volume load intravascularly from the effects of the solution is also a problem. Identification of proper controls is essential for evaluation. However, the control solution may have problems of its own that must be defined. For any proposed RBS a menu of models exploring cellular, tissue, organ and organism responses to the solution, its residual modifiers and the products of metabolism are needed to define primary and secondary effects.

Animals↗

PubMed1992

A double (exchange transfusion-carbon clearance) model for testing post-resuscitation reticuloendothelial function.

A double exchange transfusion-double carbon clearance method was evaluated for assessing reticuloendothelial (RE) function following exchange transfusion with hemoglobin solutions. Fifty percent of estimated blood volume (3% body weight) was withdrawn from anesthetized Sprague-Dawley rats and isovolumically replaced with shed blood (SB, control), lysed shed blood (LB, pos. control), human stroma-free hemoglobin solution (SFH), or polyhemoglobin solution (PHS). Thirty minutes after the exchange transfusion, colloidal carbon was injected intravenously and its vascular clearance followed for 1 hour. Then, the 50% exchange transfusion was repeated and the second carbon clearance measured. The intravascular carbon clearance constants, K, and clearance half-times, T1/2, were calculated and compared. No apparent differences in RE function were seen among the groups after the initial exchange transfusion. However, following the second exchange transfusion significant (P less than 0.05) slowing of carbon clearance was observed in lysed blood treated animals. The RE function of SFH or PHS treated animals were not different (P less than 0.05) from that of SB animals. A double exchange transfusion-double carbon clearance method seems to reveal changes in RE function that are not apparent after a single exchange transfusion and clearance test.

Animals↗

PubMed1992

Hepatic reticuloendothelial function following resuscitation with hemoglobin solutions.

Red cell substitutes could lead to depressed reticuloendothelial (RE) particulate clearance function. This hypothesis was tested using an animal model of hypovolemia-resuscitation. Anesthetized male Sprague-Dawley rats were subjected to 50% blood volume hemorrhage followed by isovolumic replacement with stroma-free hemoglobin (SFH, 7 gHb/dl), polyhemoglobin (PHS, 14 gHb/dl), or shed blood (SB). At 30 min post-transfusion, the liver was isolated and perfused with colloidal carbon. Hepatic RE function was assessed from the carbon clearance kinetics. In separate experiments, the hepatic Kupffer cells were isolated and cultured from rats that were previously hemorrhaged and transfused with normal saline solution. The cultured Kupffer cells were incubated with SFH or bovine albumin (ALB) and their phagocytic function assessed in-vitro. The hepatic carbon clearance following exchange transfusion with hemoglobin solutions was not significantly altered as compared to shed blood controls (P greater than 0.05). Similarly, phagocytic function of hemoglobin treated Kupffer cells was not significantly different (P greater than 0.05) from that of ALB treated cells.

Animals↗

PubMed1992

Coagulation dynamics after hemodilution with polyhemoglobin.

The effect of polyhemoglobin solution (PHS), a candidate erythrocyte cell substitute, on blood coagulation, was investigated. Whole blood samples from six male Sprague-Dawley rats were diluted in vitro 3:1, 1:1 and 1:3 with normal saline solution, stroma-free hemoglobin (SFH) (7 grams of hemoglobin per deciliter), PHS (14 grams of hemoglobin per deciliter) or 5 percent bovine albumin (ALB) and tested with a thrombelastograph (TEG) for effects of hemodilution on coagulation. For in vivo tests, 24 rats were randomly divided into four groups of six each and 50 percent of the estimated blood volume was replaced with SFH, PHS, ALB or 6 percent hydroxyethyl starch. Blood samples collected before and after the hemodilution were tested with a TEG. The TEG parameters (r, k, Ma, and fi) were determined and analyzed statistically. At 50 percent in vitro or in vivo hemodilution, PHS did not cause significant alteration (p greater than 0.05) in initial coagulation mechanism, while SFH slightly, but significantly (p less than 0.05), accelerated coagulation. Tensile strength of formed clot (Ma) did decrease after hemodilution with PHS (p less than 0.05), but the effect was attributable to dilutional effect. At moderate hemodilution (50 percent), PHS does not seem to cause undue coagulopathy in this rat model.

Animals↗

PubMed1991

Reconstitution of the skeletal sarcoplasmic reticulum Ca2(+)-pump: influence of negatively charged phospholipids.

The Ca2(+)-ATPase of skeletal sarcoplasmic reticulum was purified and reconstituted in the presence of phosphatidyl choline using the freeze-thaw sonication technique. The effect of incorporation of negatively charged phospholipids, phosphatidylserine and phosphatidylinositol phosphate, into the phosphatidylcholine proteoliposomes was investigated. Various ratios of phosphatidylserine or phosphatidylinositol phosphate to phosphatidylcholine were used, while the total amount of phospholipid in the reconstituted vesicles was kept constant. Enrichment of phosphatidylcholine proteoliposomes by phosphatidylserine or phosphatidylinositol phosphate was associated with activation of Ca2(+)-uptake and Ca2(+)-ATPase activities. The highest activation was obtained at a 50:50 molar ratio of phosphatidylserine:phosphatidylcholine and at a 10:90 molar ratio of phosphatidylinositol phosphate:phosphatidylcholine. The initial rates of Ca2(+)-uptake obtained at 1 microM Ca2+ were 2.6 +/- 0.1 mumol/min per mg of phosphatidylserine:phosphatidylcholine proteoliposomes and 1.5 +/- 0.1 mumol/min per mg of phosphatidylinositol phosphate:phosphatidylcholine proteoliposomes, compared to 0.9 +/- 0.05 mumol/min per mg of phosphatidylcholine proteoliposomes. These findings suggest that negatively charged phospholipids may be involved in the activation of the reconstituted skeletal muscle sarcoplasmic reticulum Ca2(+)-pump.

Animals↗

PubMed1991

Use of intravenous gamma globulin to passively immunize high-risk children against respiratory syncytial virus: safety and pharmacokinetics. The RSVIG Study Group.

Infants with cardiopulmonary disease develop severe illness from respiratory syncytial virus (RSV) infection. Safety, feasibility, and pharmacokinetics of intravenous gamma globulin (IVIG) to prevent RSV illness were studied in 23 high-risk infants in a phase I trial. IVIG with an RSV neutralizing antibody titer of 1:1,100 in 5% solution was given monthly over a 2- to 4-h period in a clinical setting during the RSV season. The first group (n = 7) received 500 mg/kg of body weight, the second group (n = 9) received 600 mg/kg, and the third group (n =7) received 750 mg/kg. Serum was drawn prior to infusion and 2, 14, and 30 days after infusion. Total immunoglobulin G and RSV A2 and RSV B neutralizing antibody levels were obtained after the first IVIG infusion. Two children developed mild reversible pulmonary edema (group receiving 600 mg/kg per dose), and one developed hives and wheezing during one infusion (group receiving 500 mg/kg per dose). Twelve children developed subsequent RSV infection during two RSV seasons (November to April) over a 2-year follow-up period; 9 of 12 developed infection during the infusion year. Eleven illnesses were mild; one child died of progressive RSV illness (group receiving 500 mg/kg per dose). A cumulative infusion effect was not observed. IVIG appears safe and feasible in an outpatient setting, and at 750 mg/kg per dose, a target RSV antibody level of greater than or equal to 1:100 was achieved.

Child, Preschool↗

PubMed1990

The role of phospholamban in the regulation of calcium transport by cardiac sarcoplasmic reticulum.

The calcium transport mechanism of cardiac sarcoplasmic reticulum (SR) is (SR) is regulated by a phosphoregulatory mechanism involving the phosphorylation-dephosphorylation of an integral membrane component, termed phospholamban. Phospholamban, a 27,000 Da proteolipid, contains phosphorylation sites for three independent protein kinases: 1) cAMP-dependent, 2) Ca2(+)-calmodulin-dependent, and 3) Ca2(+)-phospholipid-dependent. Phosphorylation of phospholamban by any one of these kinases is associated with stimulation of the calcium transport rates in isolated SR vesicles. Dephosphorylation of phosphorylated phospholamban results in the reversal of the stimulatory effects produced by the protein kinases. Studies conducted on perfused hearts have shown that during exposure to beta-adrenergic agents, a good correlation exists between the in situ phosphorylation of phospholamban and the relaxation of the left ventricle. Phosphorylation of phospholamban in situ is associated with stimulation of calcium transport rates by cardiac SR, similar to in vitro findings. Removal of beta-adrenergic agents results in the reversal of the inotropic response and this is associated with dephosphorylation of phospholamban. These findings indicate that a phospho-regulatory mechanism involving phospholamban may provide at least one of the controls for regulation of the contractile properties of the myocardium.

Animals↗

PubMed1990

The specific incorporation of labelled aromatic amino acids into proteins through growth of bacteria in the presence of glyphosate. Application to fluorotryptophan labelling to the H(+)-ATPase of Escherichia coli and NMR studies.

Growth of Escherichia coli in the presence of glyphosate, an inhibitor of aromatic amino acid biosynthesis, has permitted the production of proton translocating ATPase that is specifically labelled with 5-fluorotryptophan. Five sets of 19F nuclear magnetic resonances are resolved. The use of glyphosate should be of wide applicability in the preparation of proteins labelled in aromatic amino acid residues for NMR studies.

Amino Acids↗

PubMed1990

Functional reconstitution of the cardiac sarcoplasmic reticulum Ca2(+)-ATPase with phospholamban in phospholipid vesicles.

The Ca2(+)-ATPase in cardiac sarcoplasmic reticulum (SR) is under regulation by phospholamban, an oligomeric proteolipid. To determine the molecular mechanism by which phospholamban regulates the Ca2(+)-ATPase, a reconstitution system was developed, using a freeze-thaw sonication procedure. The best rates of Ca2+ uptake (700 nmol/min/mg reconstituted vesicles compared with 800 nmol/min/mg SR vesicles) were observed when cholate and phosphatidylcholine were used at a ratio of cholate/phosphatidylcholine/Ca2(+)-ATPase of 2:80:1. The EC50 values for Ca2+ were 0.05 microM for both Ca2+ uptake and Ca2(+)-ATPase activity in the reconstituted vesicles compared with 0.63 microM Ca2+ in native SR vesicles. Inclusion of phospholamban in the reconstituted vesicles was associated with a significant inhibition of the initial rates of Ca2+ uptake at pCa 6.0. However, phosphorylation of phospholamban by the catalytic subunit of the cAMP-dependent protein kinase reversed the inhibitory effect on the Ca2+ pump. Similar findings were observed when a peptide, corresponding to amino acids 1-25 of phospholamban, was used. These findings indicate that phospholamban is an inhibitor of the Ca2(+)-ATPase in cardiac SR and phosphorylation of phospholamban relieves this inhibition. The mechanism by which phospholamban inhibits the Ca2+ pump is unknown, but our findings with the synthetic peptide suggest that a direct interaction between the Ca2(+)-ATPase and the hydrophilic portion of phospholamban may be one of the mechanisms for regulation.

Animals↗

PubMed1990

Antibody responses of humans and nonhuman primates to individual antigenic sites of the hemagglutinin-neuraminidase and fusion glycoproteins after primary infection or reinfection with parainfluenza type 3 virus.

An unusual feature of human parainfluenza virus type 3 (PIV3) is ita ability to cause reinfection with high efficiency. The antibody responses of 45 humans and 9 rhesus monkeys to primary infection or subsequent reinfection with PIV3 were examined to identify deficiencies in host immunologic responses that might contribute to the ability of the virus to cause reinfection with high frequency. Antibody responses in serum were tested by using neutralization and hemagglutination inhibition (HI) assays and a monoclonal antibody blocking immunoassay able to detect antibodies to epitopes within six antigenic sites on the PIV3 hemagglutinin-neuraminidase (HN) glycoprotein and eight antigenic sites on the fusion (F) protein. Primary infection of seronegative infants or children with PIV3 stimulated strong and rather uniform HI and neutralizing antibody responses. More than 90% of the individuals developed antibodies to four of the six HN antigenic sites (including three of the four neutralization sites), but the responses to F antigenic sites were of lesser magnitude and varied considerably from person to person. Young infants who possessed maternally derived antibodies in their sera developed lower levels and less frequent HI, neutralizing, and antigenic site-specific responses to the HN and F glycoproteins than did seronegative infants and children. In contrast, children reinfected with PIV3 developed even higher HI and neutralizing antibody responses than those observed during primary infection. Reinfection broadened the HN and F antigenic site-specific responses, but the latter remained relatively restricted. Adults possessed lower levels of HI, neutralizing, and antigenic site-specific antibodies in their sera than did children who had been reinfected, suggesting that these antibodies decay with time. Rhesus monkeys developed more vigorous primary and secondary antibody responses than did humans, but even in these highly responsive animals, response to the F glycoprotein was relatively restricted following primary infection. Bovine PIV3 induced a broader response to human PIV3 in monkeys than was anticipated on the basis of their known relatedness as defined by using monoclonal antibodies to human PIV3. These observations suggest that the restricted antibody responses to multiple antigenic sites on the F glycoprotein in young seronegative infants and children and the decreased responses to both the F and HN glycoproteins in young infants and children with maternally derived antibodies may play a role in the susceptibility of human infants and young children to reinfection with PIV3.

Adult↗

PubMed1989

Kinetics of human erythrocyte glucose-6-phosphate dehydrogenase dimers.

The steady-state kinetics of human erythrocyte glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ 1-oxidoreductase, EC 1.1.1.49) dimers were studied by initial rate measurement. These experiments gave intersecting double-reciprocal plots suggesting a ternary complex mechanism with a Km for NADP and glucose 6-phosphate of 11 microM and 43 microM, respectively. These studies were combined with rate measurements in the presence of one product (NADPH), dead-end inhibitors, as well as alternative substrates. The inhibition by NADPH was found to be competitive with respect to both substrates. Alternate substrates experiments gave linear double-reciprocal plots over a wide range of substrate concentrations. The results suggest that the dimeric enzyme follows either a random or a Theorell-Chance mechanism.

Chromatography, High Pressure Liquid↗

PubMed1989

Testing for syphilis.

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Emergency Service, Hospital↗

PubMed1989

[Studies on newly improved gallium alloy as dental restorative material, by addition of SiC whisker and titanium powder].

Studies on new gallium alloy using liquid alloy of Ga-Sn-In which was stable at room temperature and with Ag-Sn-Cu-Pd-Zn alloy as powder, were undertaken. Two kinds of gallium alloy by addition of SiC whisker and titanium powder were made on an experimental basis for the purpose of improving the property of this alloy (reinforced alloy). Physical properties such as compressive strength, diametral tensile strength, dimensional change and chemical properties such as discoloration, corrosion, quantitative analysis by Inductively Coupled Plasma were measured. Furthermore breakage surface of alloy after tensile test was observed with SEM. The effect of adding SiC whisker and titanium powder to gallium alloy was discussed and the conclusions were as follows; 1) Compressive strength: Initial strength decreased due to addition of SiC whisker and titanium powder. However, it showed a tendency to increase as time elapsed. It approached the same degree of compressive strength after 7 days. 2) Diametral tensile strength: The best results were obtained by addition of 1% SiC whisker to gallium alloy and the optimum trituration time was 13 seconds in this study. 3) Dimensional change: Expansion in the dimensional change was observed in all testing groups which was slightly over the limitation of ISO and ADA No.1 specification. 4) Discoloration: In the case with addition of titanium powder to gallium alloy, an improvement of discoloration was manifested in artificial saliva and 1% NaS, especially prominent in the former. The favorable results were obtained by addition of 10% titanium powder to gallium alloy for discoloration tolerance. 5) Corrosion: The corrosion was low in four kinds of test immersion liquid. The best results were obtained by addition of 10% titanium powder to gallium alloy. 6) Quantitative analysis by Inductively Coupled Plasma: The examined amount of Zn dissolved out from core composition was decreased due to addition of titanium powder. An improvement of amount in dissolution of Ga, Sn and In from matrix was recognized. 7) In physical and chemical experiment's results, it was judged that the addition of 0.5-0.7% SiC whisker and 5-10% titanium powder was useful.

Copper↗

PubMed1988

Current approaches to the development of vaccines effective against parainfluenza and respiratory syncytial viruses.

Vaccines against parainfluenza (PIV) and respiratory syncytial viruses (RSV) that are currently being developed include both live and subunit vaccines. Candidate live PIV vaccines that have been found to be attenuated and efficacious in rodents or primate models are (1) cold-adapted, temperature-sensitive mutants of PIV-type 3 that have been serially passaged at low temperature (20 degrees C) in simian kidney tissue culture; (2) protease-activation mutants (PIV-1-Sendai), which have mutations that decrease the cleavability of their F glycoprotein by host cell protease; (3) an animal virus, bovine PIV-3 virus, which is antigenically related to the human PIV-3 virus, and (4) vaccinia recombinant viruses bearing RSV or PIV-3 glycoproteins. Subunit RSV and PIV-3 viruses are being produced and evaluated as immunogens. A major concern with these vaccines is the possibility of disease potentiation following virus infection as occurred previously with formalin-inactivated measles and RSV vaccines. Studies indicate that PIV-3 and RSV glycoprotein vaccines are immunogenic and efficacious in animals but insufficient data exist to estimate their capacity to potentiate disease. However, since a cotton rat model is available to detect potentiated disease resulting from infection of cotton rats previously immunized with formalin-inactivated RSV vaccine, it is now possible to systematically evaluate new vaccines in experimental animals for disease potentiation before studies are initiated in humans. It is likely within the next several years that one or more of these PIV or RSV vaccines will be tested in humans for safety and immunogenicity.

Animals↗

PubMed1988

Strain-specific serum antibody responses in infants undergoing primary infection with respiratory syncytial virus.

We sought to characterize the human respiratory syncytial virus (RSV) subgroup-specific antibody response to primary infection with RSV. RSV isolates from 43 infants and young children were typed as either subgroup A (Long strain-like) or B (18537 strain-like) based on reactivity with monoclonal antibodies. Acute-phase or preinfection and convalescent-phase sera were collected from the 43 subjects and tested in an enzyme-linked immunosorbent assay using purified F and G glycoproteins from RSV subgroups A and B and by neutralization assay against both subgroups of RSV. Twenty-six individuals had rises in titer of antibody to F glycoprotein, 27 to G glycoprotein, and 28 had rises in titer of neutralizing antibody. The antibody responses to homologous and heterologous F glycoproteins were not significantly different. In contrast, homologous vs. heterologous antibody responses to G glycoprotein were significantly different, with 7.3% relatedness between the G glycoproteins of subgroups A and B. Analysis of neutralizing antibody responses revealed 31% relatedness.

Antibodies, Monoclonal↗