• Title/Summary/Keyword: atpI

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RAPD Analysis for Genetic Diversity of Melon Species (참외와 멜론의 유전적 다양성에 대한 RAPD 분석)

  • Mo, Suk-Youn;Im, Sung-Hee;Go, Gwan-DaI;Ann, Chong-Mun;Kim, Doo Hwan
    • Horticultural Science & Technology
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    • v.16 no.1
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    • pp.21-24
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    • 1998
  • RAPD markers were analyzed in order to detect the genetic variation and diversity of the fifty-two melon lines. SDS extraction method produced more and purer DNA than CTAB method. RAPD reaction conditions were optimized as follows ; 10ng template DNA, 270nM primer, $200{\mu}M$ each of dATP, dCTP, dGTP and dTTP, $0.3{\mu}unit$ dynazyme and 10x buffer brought to $15{\mu}l$ final volume with distilled water. The adequate annealing temperature was $39^{\circ}C$ and forty cycles of amplification produced the best RAPD band patterns. Among a total of 123 bands from 12 random primers, 25 polymorphic bands(20%) were selected as reliable markers. The average number of polymorphic bands per primer was 2.1 among the 52 lines. Intragroup genetic relationship based on the marker difference was closer than intergroup genetic relationship. The 52 lines could be grouped into two major group (Korean landraces and melon lines) and then melon group subdivided into two subgroups (net melon lines and no-net melon). This result corresponded to morphological grouping. Eight RAPD markers separated the Korean landraces and melon groups and four RAPD markers separated net melon and no-net melon groups.

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A Study on the Inhibitory Effect of Yeongdamsagantang on Alzheimer in $A{\beta}-oligomer-induced$ Neuro 2A Cell Lines (($A{\beta}-oligomer$로 유도된 Neuro2A 세포주에서 용담사간탕(龍膽瀉肝湯)의 치매 억제 효과)

  • Kim, Hae-Su;Shin, Yoo-Jeong;Park, Jong-Hyuk;Kim, Seung-Mo;Paek, Kyung-Min;Park, Chi-Sang
    • The Journal of Korean Medicine
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    • v.29 no.2
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    • pp.151-164
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    • 2008
  • Objective: To investigate the effects of Yeongdamsagantang (YDGT) on apoptosis of neuronal cells that can result in dementia. Method: The water extract of the YDGT was tested in vitro for its beneficial effects on neuronal survival and neuroprotective functions, particularly in connection with $A{\beta}$ oligomer-related dementias. $A{\beta}$ oligomers derived from proteolytic processing of the ${\beta}-amyloid$ precursor protein (APP), including the $amyloid-{\beta}$ peptide $(A{\beta})$, play a critical role in the pathogenesis of Alzheimer's disease. A neuroblastoma cell line stably expressing an $A{\beta}$ oligomerassociated neuronal degeneration was used to investigate if YDGT inhibits formation of $A{\beta}$ oligomer. To measure the ATP generating level in mitochondrial membrane, luciferin/luciferase luminescence kit (Promega) and luminator was used, and to survey the protein's apparition, confocal microscopy was used. Result: $A{\beta}oligomer$ had a profound attenuation in the increase in CT105 expressing neuro2A cells from YDGT. Experimental evidence indicates that YDGT protected against neuronal damage from cells, but its cellular and molecular mechanisms remain unknown. We demonstrated that YDGT inhibited formation of $amyloid-{\beta}$ $(A{\beta})$ oligomers, which were the behavior, and possibly causative, features of AD. The decreased $A{\beta}$ oligomer in the presence of YDGT was observed in the conditioned medium of this $A{\beta}oligomer-secreting$ cell line under in vitro. In the cells, YDGT significantly attenuated mitochondrion-initiated apoptosis. Conclusion: (i) a direct $A{\beta}$ oligomer toxicity and the apoptosis initiated by the mitochondria; and (ii) multiple cellular and molecular neuroprotective mechanisms, including attenuation of apoptosis and direct inhibition of $A{\beta}$ oligomer aggregation, underlie the neuroprotective effects of YDGT.

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Sodium Dependent Taurine Transport into the Choroid Plexus, the Blood-Cerebrospinal Fluid Barrier

  • Chung, Suk-Jae;Ramanathan, Vikram;Brett, Claire M.;Giacomini, Kathleen M.
    • Journal of Pharmaceutical Investigation
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    • v.25 no.3
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    • pp.7-20
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    • 1995
  • Taurine, a ${\beta}-amino$ acid, plays an important role as a neuromodulator and is necessary for the normal development of the brain. Since de novo synthesis of taurine in the brain is minimal and in vivo studies suggest that taurine dose not cross the blood-brain barrier, we examined whether the choroid plexus, the blood-cerebrospinal fluid (CSF) barrier, plays a role in taurine transport in the central nervous system. The uptake of $[^3H]-taurine$ into ATP depleted choroid plexus from rabbit was substantially greater in the presence of an inwardly directed $Na^+$ gradient taurine accumulation was negligible. A transient in side-negative potential gradient enhanced the $Na^+-driven$ uptake of taurine into the tissue slices, suggesting that the transport process is electrogenic, $Na^+-driven$ taurine uptake was saturable with an estimated $V_{max}$ of $111\;{\pm}\;20.2\;nmole/g/15\;min$ and a $K_M\;of\;99.8{\pm}29.9\;{\mu}M$. The estimated coupling ratio of $Na^+$ and taurine was $1.80\;{\pm}\;0.122.$ $Na^+-dependent$ taurine uptake was significantly inhibited by ${\beta}-amino$ acids, but not by ${\alpha}-amino$ acids, indicating that the transporter is selective for ${\beta}-amino$ acids. Since it is known that the physiological concentration of taurine in the CSF is lower than that in the plasma, the active transport system we characterized may face the brush border (i.e., CSF facing) side of the choroid plexus and actively transport taurine out of the CSF. Therefore, we examined in vivo elimination of taurine from the CSF in the rat to determine whether elimination kinetics of taurine from the CSF is consistent with the in vitro study. Using a stereotaxic device, cannulaes were placed into the lateral ventricle and the cisterna magna of the rat. Radio-labelled taurine and inulin (a marker of CSF flow) were injected into the lateral ventricle, and the concentrations of the labelled compounds in the CSF were monitored for upto 3 hrs in the cisterna magna. The apparent clearance of taurine from CSF was greater than the estimated CSF flow (p<0.005) indicating that there is a clearance process in addition to the CSF flow. Taurine distribution into the choroid plexus was at least 10 fold higher than that found in other brain areas (e. g., cerebellum, olfactory bulb and cortex). When unlabelled taurine was co-administered with radio-labelled taurine, the apparent clearance of taurine was reduced (p<0.0l), suggesting a saturable disposition of taurine from CSF. Distribution of taurine into the choroid plexus, cerebellum, olfactory bulb and cortex was similarly diminished, indicating that the saturable uptake of taurine into these tissues is responsible for the non-linear disposition. A pharmacokinetic model involving first order elimination and saturable distribution described these data adequately. The Michaelis-Menten rate constant estimated from in vivo elimination study is similar to that obtained in the in vitro uptake experiment. Collectively, our results demonstrate that taurine is transported in the choroid plexus via a $Na^+-dependent,saturable$ and apparently ${\beta}-amino$ acid selective mechanism. This process may be functionally relevant to taurine homeostasis in the brain.

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The Effect of Higenamine on the Calcium Transport of Cardiac Sarcoplasmic Reticulum (Higenamine이 심근 Sarcoplasmic Reticulum의 칼슘운반에 미치는 영향에 관한 연구)

  • Kim, Joo-Hyun;Lee, Yung-Kyoon;Kim, Hae-Won;Kim, Myung-Suk;Park, Chan-Woong;Lim, Jung-Kyoo
    • The Korean Journal of Pharmacology
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    • v.18 no.2
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    • pp.79-87
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    • 1982
  • Higenamine(dl-demethylcoclaurine, dl-1-(4-hydroxybenzyl)-6,7-dihydroxy-1,2,3,4-tetrah-ydroisoquinoline hydrochloride), which has recently been isolated from Aconite root by Drs. Kosuge and Yokota, has known to be the main cardiotonic component of the Aconite root. The present study was undertaken to investigate the effects of Higenamine on the calcium binding and release and ATPase activity of fragmented cardiac sarcoplasmic reticulum under in vitro condition. The calcium binding and release of sarcoplasmic reticulum were measured by using the double-beam spectrophotometer and the calcium sensitive dye, murexide. In the presence of $10^{-4}{\sim}5{\times}10^{-3}M$ of Higenamine, the maximal calcium binding and the initial binding rate of porcine cardiac sarcoplasmic reticulum were inhibited dose dependently by up to 43%. However, the calcium release from cardiac sarcoplasmic reticulum, which was loaded with $Ca^{++}(50{\mu}M)$, was stimulated in dose dependent manner. When incubated in the medium of 20 mM Tris-maleate(pH 7.0), 100 mM KCl, 10 mM $MgCl_2,\;0.05mM\;CaCl_2\;and\;0.014{\sim}1\;mM\;Tris-ATP\;at\;30^{\circ}C$ in the presence of Higenamine $(10^{-4}{\sim}5{\times}10^{-3}M)$, both $Ca^{++}-and\;Mg^{++}-ATPase$ of sarcoplasmic reticulum were inhibited non-competitively by Higenamine and values of $K_i$ were 4.896 mM and 6.875 mM respectively. It is suggested from the above findings that the cardiotonic effects of Higenamine might be partially explained by the inhibition of calcium binding and the stimulation of calcium release from the sarcoplasimic reticulum which may increase the free intracellular calcium that is available in the contraction of the cardiac muscle fiber.

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Laxative Activity of the Hot-Water Extract Mixture of Hovenia dulcis Thunb. and Phyllostachys pubescens Mazel in Chronic Constipation Model SD Rats

  • Oh, Kyo-Nyeo;Kim, Yujin;Choi, Eun Jin;Lee, Hyunmi;Hong, Ji Ae;Kim, Miri;Oh, Dool-Ri;Jung, Myung-A;Park, Ro-Dong;Kim, Seong-il;Yong, Ju-seon;Lee, Hui-Seop;Ban, SangOh;Choi, Chul-yung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.649-661
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    • 2020
  • This study examined the laxative effects of hot-water extracts of Hovenia dulcis Thunb. (HD), Phyllostachys pubescens Mazel (PM), and a 2:8 mixture of both (HP) in two chronic constipation models. For the loperamide-induced constipation model, animals were divided into an untreated group, negative control group (loperamide 4 mg/kg), positive control group (bisacodyl 4 mg/kg) group, and six treatment groups (HP 100 or 400, HD 50 or 100, and PM 100 or 400 mg/kg). For the low-fiber diet-induced constipation model, animals were divided into an untreated group (normal diet), negative control group (low-fiber diet), positive control group (Agio granule, 620 mg/kg), and the same treatment groups. Fecal number, weight, fecal water content, and intestinal transit ratio were higher in the groups treated with HP, HD, and PM than in the groups treated with loperamide or low-fiber diet. Thickness of colon mucosa and muscle layers were increased in the treated groups. Colon tension increased in the HP groups, and [Ca2+]i measurements using fura-2 as an indicator showed that HP inhibits ATP-mediated Ca2+ influx in IEC-18 cells. These results showed that the HP mixture has laxative activity by increased mucin secretion and inducing contractile activity and relaxation. It may be a useful therapeutic strategy for ameliorating in chronic constipation.

Inhibitory Effects of Naeso-san on Pacemaker Potentials in Interstitial Cells of Cajal of Murine Small Intestine (생쥐 소장 카할세포의 내향성 향도잡이 전압에 미치는 내소산의 억제효과에 관한 연구)

  • Hong, Noo Ri;Ahn, Tae Seok;Park, Hyun Soo;Chae, Han;Kwon, Young Kyu;Kim, Byung Joo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.6
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    • pp.630-635
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    • 2014
  • The purpose of this study was to investigate the effects of Naeso-san in interstitial cells of Cajal (ICCs) in murine small intestine. First, we isolated ICCs from murine small intestine. After that, we cultured these cells for 1 days. The patch-clamp technique was applied on ICCs that formed network-like structures in culture (1 days). Spontaneous rhythms were routinely recorded from cultured ICCs under current-clamp conditions, and the ICCs within networks displayed more robust electrical rhythms (pacemaker potentials). To understand the relationship between Naeso-san and pacemaker activity in ICCs, we examined the effects of Naeso-san on pacemaker potentials of ICCs. In current clamp mode (I = 0), the addition of Naeso-san (10 mg/ml - 50 mg/ml) decreased the amplitude and frequency of the pacemaker potentials of ICCs in a dose dependent manner. However, these effects were blocked by intracellular $GDP{\beta}S$, a G-protein inhibitor, and glibenclamide, a specific ATP-sensitive K+ channels blocker. Pretreatment with SQ-22536, an adenylate cyclase inhibitor, did not block the Naeso-san induced effects, whereas pretreatment with ODQ, a guanylate cyclase inhibitor, or L-NAME, an inhibitor of nitric oxide (NO) synthase blocked the Naeso-san induced effects. Our findings provide insight into unraveling the modulation of Naeso-san in pacemaker potentials of ICCs and developing therapeutic agents against gastrointestinal motility disorders.

Allomyrina dichotoma larva extract attenuates free fatty acid-induced lipotoxicity in pancreatic beta cells

  • Kim, Kyong;Kwak, Min-Kyu;Bae, Gong-Deuk;Park, Eun-Young;Baek, Dong-Jae;Kim, Chul-Young;Jang, Se-Eun;Jun, Hee-Sook;Oh, Yoon Sin
    • Nutrition Research and Practice
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    • v.15 no.3
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    • pp.294-308
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    • 2021
  • BACKGROUD/OBJECTIVES: Allomyrina dichotoma larva (ADL), one of the many edible insects recognized as future food resources, has a range of pharmacological activities. In a previous study, an ADL extract (ADLE) reduced the hepatic insulin resistance of high-fat diet (HFD)-induced diabetic mice. On the other hand, the associated molecular mechanisms underlying pancreatic beta-cell dysfunction remain unclear. This study examined the effects of ADLE on palmitate-induced lipotoxicity in a beta cell line of a rat origin, INS-1 cells. MATERIALS/METHODS: ADLE was administered to high-fat diet treated mice. The expression of apoptosis-related molecules was measured by Western blotting, and reactive oxidative stress generation and nitric oxide production were measured by DCH-DA fluorescence and a Griess assay, respectively. RESULTS: The administration of ADLE to HFD-induced diabetic mice reduced the hyperplasia, 4-hydroxynonenal levels, and the number of apoptotic cells while improving the insulin levels compared to the HFD group. Treatment of INS-1 cells with palmitate reduced insulin secretion, which was attenuated by the ADLE treatment. Furthermore, the ADLE treatment prevented palmitate-induced cell death in INS-1 cells and isolated islets by reducing the apoptotic signaling molecules, including cleaved caspase-3 and PARP, and the Bax/Bcl2 ratio. ADLE also reduced the levels of reactive oxygen species generation, lipid accumulation, and nitrite production in palmitate-treated INS-1 cells while increasing the ATP levels. This effect corresponded to the decreased expression of inducible nitric oxide synthase (iNOS) mRNA and protein. CONCLUSIONS: ADLE helps prevent lipotoxic beta-cell death in INS-1 cells and HFD-diabetic mice, suggesting that ADLE can be used to prevent or treat beta-cell damage in glucose intolerance during the development of diabetes.

Utilzation of Ascidian, Holocynthia roretzi -5. Processing and Quality Evaluation of Fermented Ascidian(I)- (우렁쉥이 이용에 관한 연구 -5. 우렁쉥이 젓갈의 제조 및 품질평가( I )-)

  • LEE Kang-Ho;CHO Ho-Sung;LEE Dong-Ho;RYUK Ji-Hee;CHO Young-Je;SUH Jae-Soo;KIM Dong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.26 no.3
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    • pp.221-229
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    • 1993
  • In this study fermentation of fresh ascidian was attempted to widen the utility of ascidian. Fresh deshelled and sliced ascidians were fermented for 90days at $25^{\circ}C$ with different salt contents of 5, 10, 15 and $20\%$ (w/w) and at $5^{\circ}C$ with 5 and $10\%$ salt. Changes of such components during fermentation as free amino acids, nucleotides and the related compounds, volatile basic nitrogen(VBN), trimethyl amine(TMA), amino nitrogen and total creatinine were determined. VBN increased rapidly after 30days of fermentation at $25^{\circ}C$ while slowly in cases of fermentation at $5^{\circ}C$ and with high salt concentration. Amino nitrogen and the total creatinine also increased gradually until 45 days and 30days of fermentation, respectively, hereafter tended to decrease. ATP and ADP seemed to degrade rapidly in fresh ascidian post harvest and AMP, IMP and inosine also degraded down to hypoxanthine during fermentation. After 45days of fermentation, in the free amino acid composition of fermented ascidian were taurine, proline, glutamic acid, histidine, lysine, alanine and valine in order. The amino acids known as sweetner like prolline, lysine, alanine and glycine were in increased in fermented ascidian. The result of sensory evaluation of fermented ascidian pretreated with acid or sulfite solution showed that the peculiar taste and flavor of ascidian remained without browning for 45days fermentation at $5^{\circ}C$.

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Ecosysteme de I′Etang de Berre (Mediterranee nord-occidentale) : Caracteres Generales Physiques, Chimiques et Biologiques

  • Kim, Ki-Tai
    • Korean Journal of Environmental Biology
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    • v.22 no.2
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    • pp.247-258
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    • 2004
  • Climatological, hydrological and planktonical research studies, measurements of primary production and photosynthetic efficiency from December 1976 to December 1978 have been carried out in two brackish lakes: Lake Etang de Berre and Lake Etang de Vaine located in the French Mediterranean coast, in the region of Carry-le-Rouet located on the north-west Mediterranean near Marseilles, and in fresh water inflows from 4 Rivers (Touloubre, Durance, Arc, Durancole) to Lake Etang de Berre. Physico-chemical parameters were measured for this study: water temperature, salinity, density, pH, alcalinity, dissolved oxygen (% saturation), phosphate, nitrate, nitrite, silicate etc. Diverse biological parameters were also studied: photosynthetic pigments, phaeopigments, specific composition and biomass of phytoplankton, primary pelagic production etc. Climatical factors were studied: air-temperature, solar-radiation, evaporation, direction (including strength) of winds, precipitation and freshwater volume of the four rivers. The changes in Lake ‘Etang de Berre’ ecosystem depend on the quality of the water in the Durance River, and on the effects of seawater near the entrance of the Caronte Canal. The water quality of the lake varies horizontally and vertically as a result of atmospheric phenomena, maritime currents and tides. The distribution of water temperatures is generally heterogeneous. Southeasterly winds and the Northeasterly Mistral wind are important in the origins of circulated and mixed water masses. These winds are both frequent and strong. They have, as a result, a great effect on the water environment of Lake Etang de Berre. In theory, the annual precipitation in this region is well over eight times the water mass of the lake. The water of the Durance River flows into Lake Etang de Berre through the EDF Canal, amounting to 90% of the precipitation. However, reduction of rainfall in dry seasons has a serious effect on the hydrological characteristics of the lake. The temperature in the winter is partially caused by the low temperature of fresh water, particularly that of the Durance River. The hydrological season of fresh and brackish water is about one month ahead of the hydrological season of sea water in its vicinity. The salinity of Lake Etang de Berre runs approximately 3$\textperthousand$, except at lower levels and near the entrance to the Caronte Canal. However, when the volume of the Durance River water is reduced in the summer and fall, the salinity rises to 15$\textperthousand$. In the lake, the ratio of fresh water to sea water is six to one (6:1). The large quantities of seston conveyed by rivers, particularly the Durance diversion, strongly reduce the transparency in the brackish waters. Although the amount of sunshine is also notable, transparency is slight because of the large amount of seston, carried chiefly by Tripton in the fresh water of the Durance River. Therefore, photosynthesis generally occurs only in the surface layer. The transparency progressively increases from freshwater to open seawater, as mineral particles sink to the bottom (about 1.7kg $m^{-2}a^{-1}$ on the average in brackish lakes). The concentration of dissolved oxygen and the rate of oxygen saturation in seawater (Carry-le-Rouet) ranged from 5.0 to 6.0 $m\ell$ㆍ.$1^{-1}$, and from 95 to 105%, respectively. The amount of dissolved oxygen in Etang de Berre oscillated between 2.9 and 268.3%. The monographs of phosphate, nitrate, nitrite and silicate were published as a part of a study on the ecology of phytoplankton in these environments. Horizontal and vertical distributions of these nutriments were studied in detail. The recent diversion of the Durance River into Lake Etang de Berre has effected a fundamental change in this formerly marine environment, which has had a great impact in its plankton populations. A total of 182 taxa were identified, including 111 Bacillariophyceae, 44 Chlorophyceae, and 15 Cyanophyceae. The most abundant species are small freshwater algae, mainly Chlorophyceae. The average density is about $10^{8}$ cells $1^{-1}$ in Lake Etang de Berre, and about double that amount in Lake Etang de Vaine. Differences in phytoplankton abundance and composition at the various stations or at various depths are slight. Cell biovolume V (equivalent to true biomass), plasma volume VP (‘useful’ biomass) and, simultaneously. the cell surface area S and S/V ratio through the measurement of cell dimensions were computed as the parameters of phytoplankton productivity and metabolism. Pigment concentrations are generally very high on account of phytoplankton blooms by Cyanophyceae, Chlorophyceae and Cryptophyceae. On the other hand, in freshwaters and marine waters, pigment concentrations are comparatively low and stable, showing slight annual variation. The variations of ATP concentration were closely related to those of chlorophyll a and phytoplankton blooms only in marine waters. The carbon uptake rates ranged between 38 and 1091 mg$Cm^{-2}d^{-1}$, with an average surface value of 256 mg; water-column carbon-uptake rates ranged between 240 and 2310 mg$Cm^{-2}d^{-1}$, with an average of 810, representing 290 mg$Cm^{-2}$, per year 45 000 tons per year of photosynthetized carbon for the whole lake. Gross photosynthetic production measured by the method of Ryther was studied over a 2-year period. The values obtained from marine water(Carry-le-Rouet) ranged from 23 to 2 337 mg$Cm^{-2}d^{-1}$, with a weighted average of 319, representing about 110 gCm$^{-2}$ per year. The values in brakish water (Etang de Berre) ranged from 14 to 1778 mg$Cm^{-2}d^{-1}$, with a weighted average of 682, representing 250 mg$Cm^{-2}$ per year and 38 400 tons per year of photosynthesized carbon for the whole lake.

Management of Non-pain Symptoms in Terminally Ill Cancer Patients: Based on National Comprehensive Cancer Network Guidelines (말기암환자에서 통증 외 증상의 관리: 최신 NCCN(National Comprehensive Cancer Netweork) 권고안을 중심으로)

  • Lee, Hye Ran
    • Journal of Hospice and Palliative Care
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    • v.16 no.4
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    • pp.205-215
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    • 2013
  • Most terminally ill cancer patients experience various physical and psychological symptoms during their illness. In addition to pain, they commonly suffer from fatigue, anorexia-cachexia syndrome, nausea, vomiting and dyspnea. In this paper, I reviewed some of the common non-pain symptoms in terminally ill cancer patients, based on the National Comprehensive Cancer Network (NCCN) guidelines to better understand and treat cancer patients. Cancer-related fatigue (CRF) is a common symptom in terminally ill cancer patients. There are reversible causes of fatigue, which include anemia, sleep disturbance, malnutrition, pain, depression and anxiety, medical comorbidities, hyperthyroidism and hypogonadism. Energy conservation and education are recommended as central management for CRF. Corticosteroid and psychostimulants can be used as well. The anorexia and cachexia syndrome has reversible causes and should be managed. It includes stomatitis, constipation and uncontrolled severe symptoms such as pain or dyspnea, delirium, nausea/vomiting, depression and gastroparesis. To manage the syndrome, it is important to provide emotional support and inform the patient and family of the natural history of the disease. Megesteol acetate, dronabinol and corticosteroid can be helpful. Nausea and vomiting will occur by potentially reversible causes including drug consumption, uremia, infection, anxiety, constipation, gastric irritation and proximal gastrointestinal obstruction. Metoclopramide, haloperidol, olanzapine and ondansetron can be used to manage nausea and vomiting. Dyspnea is common even in terminally ill cancer patients without lung disease. Opioids are effective for symptomatic management of dyspnea. To improve the quality of life for terminally ill cancer patients, we should try to ameliorate these symptoms by paying more attention to patients and understanding of management principles.