• Title/Summary/Keyword: Cell State

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Synthesis and Phase Relations of Potassium-Beta-Aluminas in the Ternary System K2O-MgO-Al2O3 (K2O-MgO-Al2O3 3성분계에서 K+-β/β"-Al2O3의 합성 및 상관계)

  • Ham, Choul-Hwan;Lim, Sung-Ki;Lee, Chung-Kee;Yoo, Seung-Eul
    • Applied Chemistry for Engineering
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    • v.10 no.7
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    • pp.1086-1091
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    • 1999
  • $K^+-{\beta}/{\beta}"-Al_2O_3$ in the ternary system $K_2O-MgO-Al_2O_3$ was directly synthesized by solid state reaction. The phase formation and phase relation were carefully investigated in relation to starting composition, calcining temperature and time, and dispersion medium. The optimal synthetic condition was also examined for the formation of ${\beta}"-Al_2O_3$ phase with a maximum fraction. As a composition range, the mole ratio of $K_2O$ to $Al_2O_3$ was changed from 1:5 to 1:6.2 and the amount of MgO used as a stabilizer was varied from 4.2 wt % to 6.3 wt %. The calcining temperature was selected between $1000^{\circ}C$ and $1500^{\circ}C$. At $1000^{\circ}C$, the ${\beta}/{\beta}"-Al_2O_3$ phases began to form resulted from the combining of ${\alpha}-Al_2O_3$ and $KAlO_2$ and increased with temperature rising. All of ${\alpha}-Al_2O_3$ phase disappeared to be homogenized to the ${\beta}/{\beta}"-Al_2O_3$ phase at $1200^{\circ}C$. Near the temperature at $1300^{\circ}C$, the fraction of ${\beta}"-Al_2O_3$ phase showed a maximum value with the composition of $K_{1.67}Mg_{0.67}Al_{10.33}O_{17}$. At temperatures above $1300^{\circ}C$, the fraction of ${\beta}"-Al_2O_3$ phase decreased gradually owing to $K_2O$ loss caused by a high potassium vapor pressure, and the appropriate calcining time was about 5 hours. Acetone was more effective than distilled water as a dispersion medium for milling and mixing.

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Change of RBC Indices with Serum Iron, TIBC Level in Iron Deficiency Anemia Patients Who Treated Oriental Medicine (철결핍성 빈혈 환자에서 한약 투여에 따른 혈청철, 철포합능 및 적혈구 지표의 변화)

  • Kim, Dong-Woung;Shin, Jin-Bong;Park, Bo-Ra;Won, Jin-Hee;O, Kwang-Woo;Moon, Goo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.6
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    • pp.1579-1584
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    • 2008
  • Anemia are often detected among many patients who are treated in oriental traditional medicine hospitals in Korea through blood tests which are taken to diagnose the patients' general state, and most of them are diagnosed to have iron deficiency anemia. This study amis the effect of herbal medicine on the RBC count, hemoglobin, hematocrits and indices of red blood cells such as mean corpuscular volume(MCV), mean corpuscular hemoglobin(MCH), mean corpuscular hemoglobin concentration(MCHC), serum iron, and total iron binding capacity(TIBC) among patients who were treated in oriental medical hospital and diagnosed to have anemia through blood tests other than main symptoms. The subjects of this study were 29 patients(male 7, female 22, mean age: 56.24${\pm}$19.11years) who were chosen ones, they has below hemoglobin 12 gm/dL, serum iron 70 IU/L, MCV 80 fL on laboratory screening test when they first visits our hospital. As a result, the volume of red blood cell(RBC), hematocrit(HCT), and hemoglobin(Hb) in peripheral blood increased from $348.27{\pm}8.95{\times}10^5$ /UL, 28.82${\pm}$0.56%, 9.24${\pm}$0.22 g/dL to $375.62{\pm}9.11{\times}10^5$ /UL, 31.36${\pm}$0.68%, 10.31${\pm}$0.25 g/dL comparing before and after oriental medicine treatment. After the treatment, MCH, MCHC, and MCV also increased from 26.98${\pm}$0.84 Pg, 32.62${\pm}$0.45 g/dl, 79.26${\pm}$0.17 fL to 27.74${\pm}$0.74 Pg, 32.84${\pm}$0.33 g/dl, 84.15${\pm}$1.67 fL. There was statistically significant increase in MCH, MCV(P<0.05); however, there was no significant change in MCHC(P>0.05). The amount of iron in blood increased from 32.96${\pm}$4.20 ug/dL to 67.22${\pm}$7.18 ug/dL, which showed a statistically significant increase(P<0.05). On the other hand, TIBC decreased from 325.08${\pm}$13.97 ug/dL to 315.61${\pm}$18.79 ug/dL, which was not a statistically significant change(P>0.05). Like this, all the patients showed favorable results in anemia based on the measurements of indices of red blood cells except MCHC. According to the classification of herbal medicines, in strengthening energy group(補氣), replenishing blood group(補血), and replenishing both energy and blood group(補氣血), the volume of RBC, HCT, and hemoglobin showed significant increases. Furthermore, in herbal medicines strengthening blood(補血) and replenishing both energy and blood group(補氣血), serum iron also relatively increased compare to strengthening energy group(補氣).

Limb Salvage Surgery with Intramedullary Nailing and Cementization for the Bone Tumors of the Proximal Humerus (근위 상완골 골종양에서 골수강내 금속정과 골시멘트를 이용한 사지 구제술)

  • Kim, Han-Soo;Oh, Joo-Han;Nam, Woo-Dong;Rhie, Tae-Yon;Jeong, Jin-Young;Lee, Han-Koo;Lee, Sang-Hoon
    • The Journal of the Korean bone and joint tumor society
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    • v.6 no.2
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    • pp.53-60
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    • 2000
  • Purpose : The purpose of the current study is to evaluate the functional and oncologic results of the limb salvage surgery with intramedullary nailing and cementization in malignant bone tumors of the proximal humerus. Materials and Methods : We reviewed 18 cases of limb salvage surgery of resection and reconstruction with an intramedullary nail and cement-molded humeral head for the malignant bone tumors of the proximal humerus, which performed between August, 1992 through the April, 1998. The diagnoses included the osteosarcoma in 7 patients, chondrosarcoma in 3, the recurrent giant cell tumor in 3, metastatic tumor in 3, multiple myeloma in one patient and the one patient with malignant fibrous histiocytoma. The mean age at the time of surgery was 38 years(range, 15-73 years). The mean follow-up period was 26 months(range, 6-67 months). Results : Average functional score by ISOLS evaluation system was 21.1(70.3%). There were 3 local recurrences and 5 distant metastases. There were one case of shoulder instability and one case of deep infection. The seven patients are continuously disease free state and seven patients are alive with disease. Four patients died from the disease. Conclusion : The limb salvage surgery with intramedullary nailing and cementization in the bone tumors of the proximal humerus may be considered an option for the malignant bone tumors of the proximal humerus in selected patients.

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Composition Ratio Analysis of Transesterification Products of Olive Oil by Using Thin Layer Chromatography and Their Applicability to Cosmetics (올리브 오일의 에스터 교환반응 생성물의 TLC를 이용한 조성비 분석 및 화장품에의 응용가능성 평가)

  • Park, So Hyun;Shin, Hyuk Soo;Kim, A Rang;Jeong, Hyo Jin;Xuan, Song Hua;Hong, In Kee;Lee, Dae Bong;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.29 no.3
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    • pp.342-349
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    • 2018
  • In this study, the physicochemical properties, emulsifying capacity, moisture content and cytotoxicity of the composite material produced by transesterification reactions of the olive oil (olive oil esters) were investigated for cosmetic applications. Olive oil esters with short (S) and long (L) reaction times were studied. From the TLC-image analysis, composition ratios of the olive oil esters S were found to be 5.2, 24.1, 46.4, and 21.9% for mono-, di-, tri-glyceride, and fatty acid ethyl ester, respectively. Those of the olive oil esters L were 4.1, 24.7, 40.6, and 28.8% for mono-, di-, tri-glyceride, and fatty acid ethyl ester, respectively. The iodine value, acid value, saponification value, unsaponified matter, refractive index, and specific gravity were determined and purity tests were also carried out and normalized to establish standards and testing methods for using olive oil esters in cosmetics. To evaluate their emulsifying capacities, the O/W emulsion was prepared without surfactants and the formation of the emulsified particles were confirmed. After 5 days of applying the olive oil esters to human skin, the skin moisture retention was improved by 13.1% from the initial state. For the evaluation of toxicity on human skin cells, the olive oil esters showed 90% or more of the cell viability at $0.2-200{\mu}g/mL$. These results suggested that olive oil esters can be applied as natural/non-toxic ingredients to cosmetics industries.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Recent Advancement in the Differentiation of Tissues and Organs and Regulation of Gene Expression (조직.기관의 분화와 유전자 발현의 조절, 최근의 진보)

  • Harn, Chang-Yawl
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.1-35
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    • 1997
  • Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

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Effect of $PGE_2$ and $PGF_{2{\alpha}}$ on the Osmotic Fragility and Membrane $Ca^{++}$ Binding in Human Erythrocytes ($PGE_2$$PGF_{2{\alpha}}$가 삼투성 용혈 및 적혈구막 $Ca^{++}$결합에 미치는 영향)

  • Yeoun, Dong-Soo;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.17 no.2
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    • pp.135-142
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    • 1983
  • $PGE_2$ and $PGF_{2{\alpha}}$ are known to act similarly in a number of animal tissues. They both facilitate regression of corpus luteum(Poyser, 1972; Fuch et al, 1974; Coudert et at, 1974) and stimulate contraction of uterine muscle (Laudanski et al, 1977; Porter et al, 1979; Hollingsworth et al, 1980). It is, however, not known whether these two prostaglandins exert similar actions in osmotic fragility of erythrocytes (Rasmussen et al, 1975) and $PGF_{2{\alpha}}$ alters conformation of membrane proteins (Meyers aud Swislocki, 1974). The former effect may not be mediated through changes in c- AMP concentration in the cell, since the adenylate cyclase activity in human erythrocyte is extremely low (Rodan et al, 1976; Sutherland et al, 1962) and the latter effect implies that physical state (or fluidity) of the membrane is altered by $PGF_{2{\alpha}}$. The present study was undertaken to elucidate mechanisms of action of $PGE_2$ and $PGF_{2{\alpha}}$ on the human erythocyte membrane by examining their effects on osmotic fragility and $Ca^{++}$ binding to the membrane fragments. The results are summarized as follows: 1) $PGE_2$ and $PGF_{2{\alpha}}$ increased osmotic fragility at concentrations above $10^{11}\;M$, the effect being similar for both hormones. The concentration of NaCl for 100% hemolysis was $1/16{\sim}1/17\;M$ in the presence of $10^{11}\;M\;PGE_2$ or $PGF_{2{\alpha}}$ and 1/18 M in the absence of the hormone (control). 2) When erythrocytes were suspended in 1/15 M NaCl solution, $44.2{\pm}4.3%$ of cells were hemolyzed. Addition of $10^{12}\;M\;PGE_2$ or $PGF_{2{\alpha}}$ did not increase hemolysis. When the concentration of the hormones was increased to $10^{11}\;M$, however the degree of hemolysis increased markealy to about 80%. No further increase in hemolysis was observed at concentration of the hormones above $10^{11}\;M$. 3) The additional hemolysis due to $10^{11}\;M\;PGE_2$ and $PGF_{2{\alpha}}$ appeared to he identical regardless of absence or presence of $Ca^{++}\;(0.5{\sim}10\;mM)$ in the suspending medium. 4) In the absence of prostaglandin, the binding of $Ca^{++}$ to the erythrocyte membrane increased curvilinearly as the $Ca^{++}$ concentration increased up to 5 mM above which it leveled off. A similar dependence of $Ca^{++}$ binding on the $Ca^{++}$ concentration was observed in the presence of $10^{11}\;M\;PGE_2$ or $PGF_{2{\alpha}}$, however, the amount of $Ca^{++}$ bound at a given $Ca^{++}$ concentration was significantly higher than in the absence of the hormones. 5) As in the hemolysis, $PGE_2$ and $PGF_{2{\alpha}}$ did not affect the $Ca^{++}$ binding at a concentration of $10^{12}\;M$, but increased it by about 100% at concentration above $10^{11}\;M$. These result indicate that both tile osmotic fragility of erythrocyte and the $Ca^{++}$ binding to the erythrocyte membrane are similarly enhanced by $PGE_2$ and $PGF_{2{\alpha}}$, but these two effects are not causally related. It is, therefore, concluded that the prostaglandin-induced hemolysis is not directly associated with alterations of the $Ca^{++}$ content in the membrane.

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The effectiveness of biochemical indexes for evaluating the nutrition states of children (소아에서 영양평가로서 생화학적 지표의 유용성)

  • Kim, Jae Kwang;Jin, Hyun Seong;Han, Myung Ki;Kim, Bong Seong;Cha, Choong Hwan;Park, Kie Young
    • Clinical and Experimental Pediatrics
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    • v.52 no.2
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    • pp.167-175
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    • 2009
  • Purpose : This study has been conducted to analyze whether the biochemical nutrition indexes might be useful and effective for evaluating the nutrition states of children. Methods : We evaluated 269 children, aged 3-9 years old, who had visited Gangneung Asan Hospital for elective surgery from January 2006 to December 2007, and examined their anthropometric and preoperative laboratory data with retrospective analysis. The children were classified into underweight, normal weight, overweight, and obese groups according to body mass index (BMI). The biochemical nutrition indexes (total lymphocyte count (TLC), hemoglobin, hematocrit, serum albumin, cholesterol, et al) of each group were then analyzed statistically. Results : None of the groups showed statistically significant differences in TLC. Serum albumin decreased significantly in the underweight group. Red blood cell (RBC) count, hemoglobin, hematocrit, and serum total cholesterol in the obese group were higher than in the normal weight group. None of the groups showed statistically significant increase in mean corpuscular volume or mean corpuscular hemoglobin, and it seems that the increase of hemoglobin and RBC count in the overweight and obese groups is due to the enhancement of erythropoiesis rather than iron metabolism. However, in females, almost all nutrition indexes except albumin were statistically significantly poor. Conclusion : Serum albumin, total cholesterol, RBC count, hemoglobin, and hematocrit were useful as nutrition indexes. However, except for albumin, these indexes were significantly poor for females. More control studies are needed to confirm the effectiveness of biochemical indexes for evaluating the nutritional state of children.

Electrochemical Characteristics of LiMn2O4 Cathodes Synthesized from Various Precursors of Manganese Oxide and Manganese Hydroxide (다양한 형태 및 구조의 망간산화물 및 망간수산화물 전구체로부터 합성한 LiMn2O4양극의 전기화학적 특성 연구)

  • Lee, Jong-Moon;Kim, Joo-Seong;Hong, Soon-Kie;Lee, Jeong-Jin;Ahn, Han-Cheol;Cho, Won-Il;Mho, Sun-Il
    • Journal of the Korean Electrochemical Society
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    • v.15 no.3
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    • pp.172-180
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    • 2012
  • The $LiMn_2O_4$ cathodes for lithium ion battery were synthesized from various precursors of manganese oxides and manganese hydroxides. As the first step, nanosized precursors such as ${\alpha}-MnO_2$ (nano-sticks), ${\beta}-MnO_2$ (nano-rods), $Mn_3O_4$ (nano-octahedra), amorphous $MnO_2$(nano-spheres), and $Mn(OH)_2$ (nano-plates) were prepared by a hydrothermal or a precipitation method. Spinel $LiMn_2O_4$ with various sizes and shapes were finally synthesized by a solid-state reaction method from the manganese precursors and LiOH. Nano-sized (500 nm) octahedron $LiMn_2O_4$ showed high capacities of 107 mAh $g^{-1}$ and 99 mAh $g^{-1}$ at 1 C- and 50 C-rate, respectively. Three dimensional octahedral crystallites exhibit superior electrochemical characteristics to the other one-dimensional and two-dimensional shaped $LiMn_2O_4$ nanoparticles. After 500 consecutive charge discharge battery cycles at 10 C-rate with the nano-octahedron $LiMn_2O_4$ cathode, the capacity retention of 95% was observed, which is far better than any other morphologies studied in this work.

Staged Improvement in Awareness of Disease for Elderly Cancer Patients in Southern China

  • Li, Xing;Dong, Min;Wen, Jing-Yun;Wei, Li;Ma, Xiao-Kun;Xing, Yan-Fang;Deng, Yun;Chen, Zhan-Hong;Chen, Jie;Ruan, Dan-Yun;Lin, Ze-Xiao;Wang, Tian-Tian;Wu, Dong-Hao;Liu, Xu;Hu, Hai-Tao;Lin, Jia-Yu;Li, Zhuang-Hua;Liu, Yuan-Chao;Xia, Qing;Jia, Chang-Chang;Wu, Xiang-Yuan;Lin, Qu
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6311-6316
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    • 2015
  • Background: In mainland China, awareness of disease of elderly cancer patients largely relies on the patients' families. We developed a staged procedure to improve their awareness of disease. Materials and Methods: Participants were 224 elderly cancer patients from 9 leading hospitals across Southern China. A questionnaire was given to the oncologists in charge of each patient to evaluate the interaction between family and patients, patient awareness of their disease and participation in medical decision-making. After first cycles of treatment, increased information of disease was given to patients with cooperation of the family. Then patient awareness of their disease and participation in medical decision-making was documented. Results: Among the 224 cancer elderly patients, 26 (11.6%) made decisions by themselves and 125 (55.8%) delegated their rights of decision-making to their family. Subordinate family members tended to play a passive role in decision-making significantly. Patients participating more in medical decision-making tended to know more about their disease. However, in contrast to the awareness of disease, patient awareness of violation of medical recommendations was reversely associated with their participation in medical decision-making. Improvement in awareness of diagnosis, stages and prognosis was achieved in about 20% elderly cancer patients. About 5% participated more actively in medical decision-making. Conclusions: Chinese elderly cancer patient awareness of disease and participation in medical decision-making is limited and relies on their family status. The staged procedure we developed to improve patient awareness of disease proved effective.