Protective effect of polysaccharides isolated from the seeds of Cuscuta chinensis Lam. on 5-fluorouracil-induced intestinal mucositis in mice

Purpose: To evaluate the protective effect of Cuscuta chinensis Lam. polysaccharides (PCCL) on 5-fluorouracil-(5-FU)-induced intestinal mucositis (IM) in mice. Methods: PCCL was orally administered at a dose of 20 mg·kg­1 for 7 days and its protective effect on 5-FU-induced IM (5-FU, 50 mg·kg­1 for 5 days) was evaluated by monitoring changes in body weight, degree of diarrhea, levels of tissue inflammatory factors (tumor necrosis factor α, interleukin 6, and interleukin 1ß levels), apoptosis rates, and the expression levels of caspase-3, Bax and Bcl-2. Results: The severity of mucosal injury (as reflected by body weight changes, degree of diarrhea, height of villi, and damage to crypts) was significantly attenuated by PCCL administration. PCCL also reduced the levels of tissue inflammatory factors, the apoptosis rate, and the expression of caspase-3 and Bax, and increased Bcl-2 expression. Conclusions: PCCL administration may be significantly protective against 5-FU-induced IM by inhibiting apoptosis and regulating the abnormal inflammation associated with it.

Immunohistochemical identification of GM-CSF in testis of rats with oligozoospermia with the intervention of Cuscuta chinensis flavonoids / Identificación inmunohistoquímica de GM-CSF en testículos de ratas con oligozoospermia con la intervención de flavonoides de Cuscuta chinensis

Oligozoospermia is a common infertility disease, and the incidence rate is increasing year by year. Cuscuta chinensis is a commonly used medicine for the treatment of oligozoospermia in Chinese medicine. Flavonoids are its main component. GM-CSF is a multifunctional cytokine that plays an important role in the inflammatory response. In this paper, we performed HE staining and immunohistochemical staining on the testis of rats with oligozoospermia. We intend to study the expression changes of GM-CSF in rats with oligospermia and the effect of flavonoids on the expression of GM-CSF in testis of rats with oligozoospermia.

La oligozoospermia es una enfermedad común de infertilidad, con una tasa de incidencia que aumenta año tras año. Cuscuta chinensis es un medicamento de uso común para el tratamiento de la oligozoospermia en la medicina china. Los flavonoides son su componente principal. GM-CSF es una citocina multifuncional que tiene un rol importante en la respuesta inflamatoria. En este trabajo, realizamos tinción con hematoxilina y eosina y tinción inmunohistoquímica en testículos de ratas con oligozoospermia. TNuestro objetivo fue estudiar los cambios de expresión de GM-CSF en ratas con oligozoospermia y el efecto de los flavonoides en la expresión de GM-CSF en testículos de ratas con oligozoospermia.

Development and Validation of HPLC-PDA Method and Pattern Recognition Analyses Using Eight Marker Compounds for the Quality Control Between the Seeds of Cuscuta chinensis Lam. and Cuscuta japonica Choisy

Cuscuta chinensis Lam. and Cuscuta japonica Choisy are parasitic plants. C. chinensis seeds were traditionally used for treatment of kidney and liver deficiencies. C. japonica seeds were used as tonic medicine to improve liver function and strengthen kidneys, treatment of high blood pressure, chronic diarrhea, and sore eyes. Cuscutae Semen are seeds of only C. chinensis in Korean Herbal Pharmacopoeia (K.H.P.). The developed HPLC-PDA method easily, accurately, and sensitively quantified using eight marker compounds [hyperoside (1), astragalin, (2), quercetin (3), kaempferol (4), chlorogenic acid (5), 3,4-di-O-caffeoylquinic acid (6), 1,5-di-O-caffeoylquinic acid (7), and 4,5-di-O-caffeoylquinic acid (8)]. In addition, the method may be used to distinguish seeds between C. chinensis Lam. and C. japonica Choisy. Furthermore, the result from the current study was applied to clarify samples between steam processed and unprocessed samples of C. chinensis by pattern analysis.

Beneficial effects of Cuscuta chinensis extract on glucocorticoid-induced osteoporosis through modulation of RANKL/OPG signals

Cuscuta chinensis Lam. (Convolvulaceae) is an important herbal medicine widely used to improve sexual function, treat osteoporosis, and prevent aging, and has been reported to exhibit anti-osteoporotic effects in vitro. However, the activity of Cuscuta chinensis Lam. on glucocorticoid-induced osteoporosis still remains unclear. The present study aimed to assess the protective effect and the underlying mechanism of action of Cuscuta chinensis extract (CCE) against glucocorticoid-induced osteoporosis in vivo. Sprague-Dawley rats were randomly divided into four groups as follows: control group, osteoporosis group, and 2 CCE-treated osteoporosis groups (100 mg·kg-1·day-1). Blood samples and femur bones were collected for immunohistochemistry, biochemical, mRNA expression, and western blot analysis. HPLC analysis revealed that chlorogenic acid, quercetin, and hyperin were the major constituents of CCE. The results indicated that CCE increased bone length, bone weight, and bone mineral density and suppressed dexamethasone (DEX)-induced reduction in body weight. In addition, TRAP staining indicated that CCE reduced osteoclasts in DEX-induced osteoporosis rats. Mechanistically, CCE treatment alleviated the increase of bone resorption markers and the decline of osteogenic markers, which might be partially mediated by regulation of RANKL/OPG and RunX2 pathways. These results suggest that CCE showed promising effects in the protection against glucocorticoid-induced osteoporosis through protecting osteoblasts and suppressing osteoclastogenesis.

Study on mechanism of Cuscutae Semen flavonoids in improving reproductive damage of Tripterygium Glycosides Tablets in rats based on high-throughput transcriptome sequencing / 中国中药杂志

Tripterygium Glycosides Tablets has good anti-inflammatory and immunomodulatory activities,but its reproductive damage is significant. Previous studies of the research group have found that Cuscutae Semen flavonoids can improve spermatogenic cell damage caused by Tripterygium Glycosides Tablets by regulating spermatogenic cell cycle,apoptosis and related protein expression,but the mechanism of action at the gene level is still unclear. In this study,Illumina high-throughput sequencing platform was applied in transcriptional sequencing of spermatogenic cells of rats after the intervention of Cuscutae Semen flavonoids and Tripterygium Glycosides Tablets. Differentially expressed genes were screened out and the GO enrichment and KEGG pathway analysis of differentially expressed genes were conducted to explore the mechanism of Cuscutae Semen flavonoids in improving reproductive injury caused by Tripterygium Glycosides Tablets. The results showed that 794 up-regulated genes and 491 down-regulated genes were screened in Tripterygium Glycosides Tablets group compared with the blank group. Compared with Tripterygium Glycosides Tablets,440 up-regulated genes and 784 down-regulated genes were screened in the Cuscutae Semen flavonoids+Tripterygium Glycosides Tablets group. Among them,the gene closely related to reproductive function is DNMT3 L. Analysis of GO function and KEGG signaling pathway enrichment showed that the above differentially expressed genes were mainly enriched in cell,cell process,catalytic activity,binding,ovarian steroid synthesis,thyroid hormone and other functions and pathways. The thyroid hormone signaling pathway was the common enrichment pathway of the two control groups. In a word,Cuscutae Semen flavonoids has a good treatment effect on male reproductive damage caused by Tripterygium Glycosides Tablets. The mechanism may be closely related to up-regulation of DNMT3 L genes and intervention of thyroid hormone signaling pathway. At the same time,the discovery of many different genes provides valuable information for study on the mechanism of Cuscutae Semen flavonoids and Tripterygium Glycosides Tablets compatibility decreasing toxicity and increasing efficiency.

Improved antimelanogenesis and antioxidant effects of polysaccharide from Cuscuta chinensis Lam seeds after enzymatic hydrolysis

Cuscuta chinensis polysaccharide (CPS) was extracted using hot water and enzymatically hydrolyzed C. chinensis polysaccharide (ECPS) was produced by the mannase enzymatic hydrolysis process. The purpose of this research was to investigate the antimelanogenic activity of ECPS and CPS in B16F10 melanoma cells. The in vitro antioxidant activity was assessed by their ferric iron reducing power and DPPH free radical scavenging activities. The molecular mass distribution of polysaccharides was determined using SEC-MALLS-RI. CPS was successfully enzymatically degraded using mannase and the weighted average molecular weights of CPS and ECPS were 434.6 kDa and 211.7 kDa. The results of biological activity assays suggested that the enzymatically hydrolyzed polysaccharide had superior antimelanogenic activity and antioxidant effect than the original polysaccharide. ECPS exhibited antimelanogenic activity by down-regulating the expression of tyrosinase, MITF, and TRP-1 without cytotoxic effects in B16F10 melanoma cells. In conclusion, ECPS have the potential to become a skin whitening product.

Essential oil composition, carotenoid profile, antioxidant and antimicrobial activities of the parasitic plant Cuscuta mitraeformis / Composición del aceite esencial, perfil de carotenoides, actividades antioxidantes y antimicrobianas de la planta parásita Cuscuta mitraeformis

The chemical composition of the essential oil and carotenoid content of the parasitic plant Cuscuta mitraeformis are described for the first time. The essential oil was analyzed by GC-FID and GC-MS revealing nonanal (24.6%) as the main constituent followed by thymol (16.5%) and eugenol (7.5%). The total carotenoid content (130 mg 100 g-1 FW) was determined by HPLC-DAD. The carotenoid fraction contained ß-carotene (76.4 mg 100 g-1 FW) and lutein (18.9 mg 100 g-1 FW) as the most abundant compounds. A weak antioxidant activity was observed by the essential oil against DPPH radical (IC50, 1.4 mg mL-1), whereas a strong antioxidant activity was determined for the carotenoid fraction (IC50, 60.1 µg mL-1). The essential oil inhibited the growth of Clavibacter michiganensis, Pseudomonas syringae pv. tomato and Erwinia carotovora with minimum inhibitory concentrations of 122.5, 184.5, 234.2 µg mL-1, respectively.

La composición química del aceite esencial y el contenido de carotenoides de la planta parásita Cuscuta mitraeformis se describen por primera vez. El aceite esencial fue analizado por GC-FID y GC-MS siendo el nonanal (24.6%) el constituyente principal seguido del timol (16.5%) y el eugenol (7.5%). El contenido total de carotenoides (130 mg 100 g-1 PF) fue determinado por HPLC-DAD. La fracción de carotenoides contuvo ß-caroteno (76.4 mg 100 g-1 PF) y luteína (18.9 mg 100 g-1 PF) como compuestos mayoritarios. Fue observada una actividad antioxidante débil por parte del aceite esencial frente al radical DPPH (IC50, 1.4 mg mL-1), mientras que una fuerte actividad antioxidante fue determinada para la fracción de carotenoides (IC50, 60.1 µg mL-1). El aceite esencial inhibió el crecimiento de Clavibacter michiganensis, Pseudomonas syringae pv. tomato y Erwinia carotovora con una concentración mínima inhibitoria de 122.5, 184.5, 234.2 µg mL-1, respectivamente.

Effect of the semen extract of Cuscuta chinensis on inflammatory responses in LPS-stimulated BV-2 microglia / 中国天然药物

AIM@#To investigate the anti-inflammatory activities of the semen extract of Cuscuta chinensis Lam. (Cuscutae Semen; CS) on the production of inflammatory mediators, nitric oxide (NO), prostaglandin 2 (PGE2), and proinflammatory cytokines in lipopolysaccharide (LPS)-stimulated BV-2 microglia.@*METHOD@#BV-2 cells were treated with CS extract for 30 min, and then stimulated with LPS or without for 24 h. The levels of NO, PGE2 and proinflammatory cytokines were measured by Griess assay and ELISA. The expression of inducible nitric oxide synthase (iNOS), and cyclooxygenase (COX)-2 mRNA and protein was determined by RT-PCR and Western blot, respectively. The phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK), and the nuclear expression of nuclear factor (NF)-κB p65 were investigated by Western blot analysis.@*RESULTS@#CS extract significantly decreased the production of NO and PGE2 by suppressing the expression of iNOS and COX-2 in activated microglia. CS extract decreased the production of TNF-α, IL-1β, and IL-6 by down-regulating their transcription levels. In addition, CS extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of NF-κB p65 in activated microglia.@*CONCLUSION@#These results indicate that CS extract is capable of suppressing the inflammatory response by microglia activation, suggesting that CS extract has potential in the treatment of brain inflammation.

Preliminary Report on the Safety of a New Herbal Formula and Its Effect on Sperm Quality

PURPOSE: Male infertility is a serious problem, and its prevalence has been increasing. Therefore, we investigated the safety of a new herbal formula and its effects on sperm quality. MATERIALS AND METHODS: An in vitro cytotoxicity test in TM3 Leydig cells was performed to evaluate cell viability after administration of five types of herbs separately and of a new herbal formula containing these five. An in vivo test in male mice was performed to evaluate the influence of the new herbal formula on the reproductive organs and sperm quality. After the 8- and 28-day oral administration of the new herbal formula, the weights of the reproductive organs were measured and the sperm count and motility were evaluated. RESULTS: In the in vitro cytotoxicity test, less than 80% cell viability at concentrations of 500 mg/L and 1,000 mg/L of Rubus coreanus Miquel and Cuscuta chinensis Lam was observed. However, more than 80% cell viability was observed at all the tested concentrations of the new herbal formula. After the 8- and 28-day oral administration, there were no considerable changes in body weight. The weights of the testes, epididymis, and seminal vesicles after the 8- and 28-day oral administration were similar to those of the control. The sperm count and activity were significantly improved compared with those of the control group at 8 and 28 days after 100, 200, and 400 mg of oral administration. CONCLUSIONS: The safety of the new formula and its positive effect on the sperm quality were observed after the oral administration of the formula.

Micromorphological comparative identification between several Chinese herbal medicines and their counterfeits / 中国中药杂志

<p><b>OBJECTIVE</b>To identify comparatively several commercial Chinese herbal medicines and their counterfeits.</p><p><b>METHOD</b>The micromorphological characters were identified. The shape, surface, section and other characters of the medicinal materials were identified by using anatomical lens and scanning apparatus. Pictures were taken and saved.</p><p><b>RESULT</b>Main micromorphological differences between several Chinese herbal medicine including Lonicera macranthoides, L. similis, Cuminum cyminum, Plantago asiatica, Cuscuta chinensis, Sinapis alba, Salvia miltiorrhiza and their counterfeits were identified.</p><p><b>CONCLUSION</b>The reference for the authenticity identification of Chinese herbal medicine and helpful experiences for the research of the same subject were provided.</p>

Reason for dormancy of Cuscuta chinensis seed and solving method / 中国中药杂志

<p><b>OBJECTIVE</b>To study the reason for the deep dormancy of the aged Cuscuta chinensis seed and find the solving method.</p><p><b>METHOD</b>The separated and combined treatments were applied in the orthogonal designed experiments.</p><p><b>RESULT</b>The aged seed had well water-absorbency; the water and ethanol extracts of the seeds showed an inhibition effect on germination capacity of the seeds.</p><p><b>CONCLUSION</b>The main reason for the deep dormancy of aged C. chinensis seed is the inhibitors existed in seed. There are two methods to solve the problem. The seeds is immersed in 98% of H2SO4 for 2 min followed by 500 mg x L(-1) of GA3 treatment for 60 min, or in 100 mg x L(-1) of NaOH for 20 min followed by 500 mg x L(-1) of GA3 treatment for 120 min.</p>

Notes on the flora of Yemen. 1: morphological and anatomical studies on Cuscuta planiflora Tenore, [Cuscutaceae] collected from Jabal An-Nahi Shn'ayb, Sanaa, Yemen with the reference of its phytogeographical distribution

Cuscuta planiflora is a parasitic plant on numerous plants from different families such as Lamiaceae [Thymus laevigaus], Acanthaceae [Hypoectes forsskalii] and Dipsaceae [Pteracephalus frutescens]. Morphological, anatomical and phytogeographical studies were carried on the Cuscuta planiflora which was collected from Jabal An-Nabi' Shu'ahyb located 30 km west of Sanaa the capital of Yemen, with coordinates [043° 58[-]-043° 59 E and 15° 14[-]-15° 15[-] N]. the morphological study includes stem morphology and floral structure. Anatomically, the cross, lateral section of the stem and lateral section of the haustoria's area, were studied, Phytogeographical studies were done by studying its distribution in Yemen and in plant geographical regions

Antibacterial activity of metanolic extract of 12 herbal species on 6 bacterial strains using cylinder-plate method

Resistance to antibacterial agents and sensivity reaction to such chemical compounds are the main reasons for investigators to develop new antibiotics from herbal sources. Antimicrobial effects of Metanolic extract of 12 herbal species Pistacia atlantica, Amygdalus scoparia, Cuscuta epthymum, Ephedra procera, Salsola boryosm, Sameraria armena, Scabiosa olivier, Scorzonera tortuosissima, Lepyrodielis holosteoidea, Vaccaria pyramidata, Sonchus oleraceam and Silene conoidema on 6 bacterial strains Staphylococcus aureas, Staphylococcus epidermidis, Echerichia colt, Kelebsiella pneumonia, Pseudomonas aeroginos and Bacillus subtilis were, separately, studied using Cylinder-plate method. In this laboratory study, methanolic extracts of herbal strains were prepared by maceration and after concentrating the extracts were dried. Then the concentrations of 50, 25, 12.5, 6.25 and 3.125 mg/ml of the extracts were prepared using 1:1 solution of DMSO/methanol. The standard bacteria with certain concentration [0.5 MacFarland] were inoculated on to the Muller-Hinton agar medium. Prepared extracts were dropped into cylinders and 18-24 hours after incubation and penetration of extract into the culture medium, the antibacterial effects and growth inhibitory zone [mm] were measure and values were expressed as [Mean +/- SEM]. The least and the most amount of effective concentration were 3.125 mg/ml and 50 mg/ml, respectively. The most inhibitory diameter belonged to methanolic extract of Amygdalus scoparia on Staphylococcus epidermidis which was equal to 19 +/- 0.3mm. Metanolic extracts of herbal species of Pistacia atlantica, Amygdalus scoparia and Ephedra procera had the most antibacterial effects compared to gentamicin as positive control. Regarding the side effects of the synthetic drugs and also benefits of such herbal extracts, extracts of these herbs as antibacterial agents after further investigations seems to be useful

Protection of extract from Cuscuta japonica on human sperm acrosome and ultrastructure / 中国中药杂志

<p><b>OBJECTIVE</b>To study the oxidation damage of active oxygen (ROS) to human sperm acrosome and ultrastructure, and study the function mechanism about Cuscuta japonica treating male's infertility and asthenoospermia.</p><p><b>METHOD</b>By using the Percoll gradient centrifugation, the sperm with normal physiological function were selected for the normal sperm model, and the sperm suspension were divided into the normal group, the model group, the positive control group (Vitamin C group), and the lugh, the median and the low dose gvoups of C. japonica. The ROS made from hypoxanthine-xanzine xanzine(HX-XO) and different content (0.125, 0.25, 0.5 g x mL(-1)) of extract were incubated with sperm in the oxygen environment. The acrosomic integrity rate were calculated and the sperm acrosome and ultrastructure were observed.</p><p><b>RESULT</b>The content (0.125, 0.5 g x mL(-1)) of extract had no obvious difference as compared with Vitamin C (0.25 mg x mL(-1)) in protecting the acrosome and ultrastructure, but the content (0.25 mg x mL(-1)) of extract was significantly better than Vit C (P < 0.001).</p><p><b>CONCLUSION</b>The suitable content of extract from C. japonica can significantly protect the sperm membrane, the acosomic structure and the mitochondrion function from the damage caused by ROS.</p>

Effect of Tujian decoction on protein kinase C activity in renal cortex in diabetic rat / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate influence of administration of Tujian decoction (Chinese herbal medicine) on protein kinase C (PKC) activity, renal function and structure in diabetic rat kidney.</p><p><b>METHOD</b>Experimental diabetic nephropathy model was induced by nephrectomy combined with streptozotocin (STZ) injection in sprague-dawley rat. Tujian decoction (20 g x kg(-1) x d(-1)) and Valsartan (20 mg x kg(-1) x d(-1)) were orally administrated respectively for 12 weeks. PKC activity was measured by [3H]phorbol 12, 13-dibutyrate ([3H]PDBu) binding assay. 24 h urine protein excretion (Upro) and renal pathological changes were observed.</p><p><b>RESULT</b>In 12th week, diabetic nephrectomized rats developed proteinuria, glomerulosclerosis, increased membrane PKC activity (mPKC), decreased cytosol PKC (cPKC), and increased ratio of mPKC and cPKC (M/C). Administration of Tujian decoction or Valsartan led to a reduction in proteinuria, structural injury, mPKC and M/C, and a recovery in cPKC.</p><p><b>CONCLUSION</b>Tujian decoction possesses a renoprotective effect on diabetic nephrectomized rat, at least partially via the inhibition of PKC activation in renal cortex.</p>

Anti-steroidogenic activity of methanolic extract of Cuscuta reflexa roxb. stem and Corchorus olitorius Linn. seed in mouse ovary.

Methanolic extract (ME) of both C. reflexa stem and C. olitorius seed arrested the normal oestrus cycle of adult female mouse and significantly decreased the weight of ovaries and uterus. The cholesterol and ascorbic acid contents in ovaries were significantly increased in the treated mice. Two key enzymes, delta5-3beta-hydroxysteroid dehydrogenase and glucose-6-phosphate dehydrogenase, were decreased significantly in ME of both C. reflexa stem and C. olitorius seed after 17 days of treatment. High level of substrates and low level of enzymes indicate the inhibition of steroidogenesis in treated mice and may be due to the presence of flavonoids.

Effect of qianlie huichun capsular on Fas expression and cell apoptosis of prostate gland tissue in rat / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of Qianlie Huichun capsular on Fas expression and cell apoptosis of prostate gland tissue in the model rat.</p><p><b>METHOD</b>Hypertrophy of prostate model rat was established by injecting testosterone to gelding male rats. After being treated with Qianlie Huichun ig 30 days, the rats were killed and prostate glands were resected for examination. The Fas expression was examined by immunobistochemical SABC. The cell apoptosis and the peak of cell apoptosis in the prostate gland of the rats were examined by flow cytometry. Compared with model group, the weight of prostage gland tissue in groups treated with Qianlie Huichun capsular was light (P < 0.01).</p><p><b>RESULT</b>Compared with model group, the Fas expression in all treatment groups increase(P < 0.05, P < 0.01). Compared with normal and model groups, cell apoptosis in all treatment groups increase at different level(P < 0.01).</p><p><b>CONCLUSION</b>The Qianlie Huchun capsular increases the Fas expression and cell apoptosis of model rats, and shows a definite treatment effect on the hypertrophy of prostate by promoting the apoptosis of prostate cell.</p>

Cuscuta hyalina Roth., an insect development inhibitor against common house mosquito Culex quinquefasciatus Say.

A preliminary screening of crude acetone extract of Cuscuta hyalina Roth. was conducted against the laboratory reared preadult stages of common house mosquito Culex quinquefasciatus (Say) (Diptera: Culicidae). 24 h LC50 of third and fourth instar larvae and pupae were 303 ppm, 306.44 ppm and 97.66 ppm respectively. The extract completely restrained the adult mosquito emergence at sublethal doses (50 ppm for third instar and 75 ppm for fourth instar), by disrupting the normal morphogenesis. The extract was found to be an effective oviposition deterrent at 80 ppm. The plant was also found to be effective in suppressing the adult emergence when directly sprinkled as dry powder on the water surface.

Studies on chemical constituents of Cuscuta chinensis / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of the seeds of Cuscuta chinensis.</p><p><b>METHOD</b>The separation was carried out by polyamide and silica gel chromatography, and the compounds were identified by means of physico-chemical and spectroscopic methods.</p><p><b>RESULTS</b>Eight compounds were isolated from the plant and identified as quercetin 3-O-beta-D-galactoside-7-O-beta-D-glucoside (I), quercetin 3-O-beta-D-apiofuranosyl-(1-->2)-beta-D-galactoside (II), hyperoside (III), isorhamnetin (IV), kaempferol (V), quercetin (VI), d-sesamin (VII) and 9(R)-hydroxy-d-sesamin (VIII).</p><p><b>CONCLUSION</b>Compounds IV and VII were isolated from Cuscuta for the first time, and I, II and VIII were characteristic constituents for this vegetable drug.</p>

Blue light-sensitive plasma membrane bound exogenous NADH oxidase in Cuscuta reflexa.

Protoplasts isolated from Cuscuta reflexa exhibited a higher rate of exogenous NADH oxidation as compared to NADPH in the dark. NAD(P)H oxidation was monitored by measuring the rate of oxygen consumption and this oxidase system was sensitive to blue light. Both NADH oxidase and its blue light sensitivity were inhibited by -SH group reacting agents. The corresponding changes occurring in H+-extrusion activity and intracellular ATP levels were also monitored. Stimulation of NADH oxidation under blue light corresponded to increased rate of H+-extrusion and intracellular ATP level, the converse was also true under NADH oxidase inhibitory conditions. These observations suggested a close functional association between blue light-sensitive plasma membrane bound redox activity and H+-ATPase in this tissue. Further, concanavalin A binding of protoplasts resulted in a loss in NADH oxidase activity and its blue light sensitivity suggesting apoplastic location and glycoprotein nature of the blue light sensitive NADH oxidase system in Cuscuta.