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PubMed2005

Valerian-hops combination and diphenhydramine for treating insomnia: a randomized placebo-controlled clinical trial.

CONTEXT: Insomnia is a prevalent health complaint associated with daytime impairments, reduced quality of life, and increased health-care costs. Although it is often self-treated with herbal and dietary supplements or with over-the-counter sleep aids, there is still little evidence on the efficacy and safety of those products. OBJECTIVE: To evaluate the efficacy and safety of a valerian-hops combination and diphenhydramine for the treatment of mild insomnia. DESIGN AND SETTING: Multicenter, randomized, placebo-controlled, parallel-group study conducted in 9 sleep disorders centers throughout the United States. PATIENTS: A total of 184 adults (110 women, 74 men; mean age of 44.3 years) with mild insomnia. INTERVENTIONS: (1) Two nightly tablets of standardized extracts of a valerian (187-mg native extracts; 5-8:1, methanol 45% m/m) and hops (41.9-mg native extracts; 7-10:1, methanol 45% m/m) combination for 28 days (n = 59), (2) placebo for 28 days (n = 65), or (3) 2 tablets of diphenhydramine (25 mg) for 14 days followed by placebo for 14 days (n = 60). OUTCOME MEASURES: Sleep parameters measured by daily diaries and polysomnography, clinical outcome ratings from patients and physicians, and quality of life measures. RESULTS: Modest improvements of subjective sleep parameters were obtained with both the valerian-hops combination and diphenhydramine, but few group comparisons with placebo reached statistical significance. Valerian produced slightly greater, though nonsignificant, reductions of sleep latency relative to placebo and diphenhydramine at the end of 14 days of treatment and greater reductions than placebo at the end of 28 days of treatment. Diphenhydramine produced significantly greater increases in sleep efficiency and a trend for increased total sleep time relative to placebo during the first 14 days of treatment. There was no significant group difference on any of the sleep continuity variables measured by polysomnography. In addition, there was no alteration of sleep stages 3-4 and rapid eye movement sleep with any of the treatments. Patients in the valerian and diphenhydramine groups rated their insomnia severity lower relative to placebo at the end of 14 days of treatment. Quality of life (Physical component) was significantly more improved in the valerian-hops group relative to the placebo group at the end of 28 days. There were no significant residual effects and no serious adverse events with either valerian or diphenhydramine and no rebound insomnia following their discontinuation. CONCLUSIONS: The findings show a modest hypnotic effect for a valerian-hops combination and diphenhydramine relative to placebo. Sleep improvements with a valerian-hops combination are associated with improved quality of life. Both treatments appear safe and did not produce rebound insomnia upon discontinuation during this study. Overall, these findings indicate that a valerian-hops combination and diphenhydramine might be useful adjuncts in the treatment of mild insomnia.

Adult↗
PubMed2003

Characterization of a plant homolog of hop, a cochaperone of hsp90.

The 90-kD molecular chaperone hsp90 is the key component of a multiprotein chaperone complex that facilitates folding, stabilization, and functional modulation of a number of signaling proteins. The components of the animal chaperone complex include hsp90, hsp70, hsp40, Hop, and p23. The animal Hop functions to link hsp90 and hsp70, and it can also inhibit the ATPase activity of hsp90. We have demonstrated the presence of an hsp90 chaperone complex in plant cells, but not all components of the complex have been identified. Here, we report the isolation and characterization of soybean (Glycine max) GmHop-1, a soybean homolog of mammalian Hop. An analysis of soybean expressed sequence tags, combined with preexisting data in literature, suggested the presence of at least three related genes encoding Hop-like proteins in soybean. Transcripts corresponding to Hop-like proteins in soybean were detected under normal growth conditions, and their levels increased further in response to stress. A recombinant GmHop-1 bound hsp90 and its binding to hsp90 could be blocked by the tetratricopeptide repeat (TPR) domain of rat (Rattus norvegicus) protein phosphatase 5. Deletion of amino acids 325 to 395, adjacent to the TPR2A domain in GmHop-1, resulted in loss of hsp90 binding. In a minimal assembly system, GmHop-1 was able to stimulate mammalian steroid receptor folding. These data show that plant and animal Hop homologs are conserved in their general characteristics, and suggest that a Hop-like protein in plants is an important cochaperone of plant hsp90.

Amino Acid Sequence↗
PubMed2002

Membrane-bound ATPase contributes to hop resistance of Lactobacillus brevis.

The activity of the membrane-bound H+-ATPase of the beer spoilage bacterium Lactobacillus brevis ABBC45 increased upon adaptation to bacteriostatic hop compounds. The ATPase activity was optimal around pH 5.6 and increased up to fourfold when L. brevis was exposed to 666 microM hop compounds. The extent of activation depended on the concentration of hop compounds and was maximal at the highest concentration tested. The ATPase activity was strongly inhibited by N,N'-dicyclohexylcarbodiimide, a known inhibitor of FoF1-ATPase. Western blots of membrane proteins of L. brevis with antisera raised against the alpha- and beta-subunits of FoF1-ATPase from Enterococcus hirae showed that there was increased expression of the ATPase after hop adaptation. The expression levels, as well as the ATPase activity, decreased to the initial nonadapted levels when the hop-adapted cells were cultured further without hop compounds. These observations strongly indicate that proton pumping by the membrane-bound ATPase contributes considerably to the resistance of L. brevis to hop compounds.

Adenosine Triphosphatases↗
PubMed1996

Elastic and length-force characteristics of the gastrocnemius of the hopping mouse (Notomys alexis) and the rat (Rattus norvegicus).

The aim of this study was to compare the contractile and series elastic properties of terrestrial mammals that use bipedal versus quadrupedal gaits. The gastrocnemius muscle of the hopping mouse (body mass 30.2 +/- 2.4 g, mean +/- S.D.) and the rat (313 +/- 10.7 g) were compared with data from the literature for the wallaby and the kangaroo rat to distinguish scaling effects and locomotion-related effects on muscle properties. Contractile length-force properties and series elastic stiffness were measured in situ during maximal tetanic contractions. The rat had a larger muscle-fibre-to-tendon-length ratio. The rat and hopping mouse showed similar normalised length-force characteristics of the gastrocnemius. Normalised stiffness in the hopping mouse was higher. The hopping mouse showed a higher capacity to store elastic energy per unit of contractile work capacity, as well as per unit of body mass. Accounting for body size differences, the rat had a smaller relative muscle mass and thus smaller work capacity than the three hopping animals considered. This is an agreement with a quadrupedal versus bipedal locomotion style. The differences in contractile and elastic properties of the gastrocnemius of the rat and hopping mouse seem to be closely related to locomotion patterns. Small animals seem to be able to utilise the storage and release of elastic energy to a far lesser extent than larger animals. However, even in animals as small as hopping mice, the storage and utilisation of elastic energy during locomotion is of functional significance and probably depends on locomotor behaviour.

Animals↗
PubMed2001

Hop tests as predictors of dynamic knee stability.

Single leg hop tests are commonly used as physical performance measures of function and are also commonly used to evaluate progress in knee rehabilitation programs, particularly for individuals recovering from anterior cruciate ligament injury or reconstructive surgery. While there is some evidence that hop tests may show promise as a predictive measure for identifying individuals who are at risk for recurrent dynamic instability, further work is needed to clearly define the role of hop test measurements for this purpose. The purposes of this clinical commentary are to review the research that has been done to establish hop tests as a physical performance measure of function, to discuss neuromuscular and biomechanical considerations related to hop performance and dynamic knee stability, to discuss existing evidence that supports the potential for hop tests as a predictor of dynamic knee stability, and to discuss considerations for future studies that are designed to more clearly define the role of hop tests in predicting dynamic knee stability.

Anterior Cruciate Ligament Injuries↗
PubMed2004

Exposure of hop growers to bioaerosols.

Air sampling was performed during picking and sorting of hop (Humulus lupulus) cones on 19 hop farms located in eastern Poland. The concentration and composition of airborne microflora and the concentration of airborne dust and endotoxin were determined. Additionally, 7 samples of settled hop dust were collected and examined for the presence of microorganisms and endotoxin. Total concentrations of airborne microorganisms were within a range of 2.08-129.58 x 10(3) cfu/m(3). Gram-positive bacteria formed 22.2-96 % of the total count. Among them, prevailed corynebacteria and endospore-forming bacilli. Fungi constituted 3.7-65.4 % of the total count. The dominant species were Penicillium citrinum, Alternaria alternata, and Cladosporium epiphyllum. Thermophilic actinomycetes and Gram-negative bacteria were detected in the air of only 10 and 6 farms, respectively. Airborne dust concentrations at the workplace ranged from 0.17-31.67 mg/m(3). The concentrations of airborne endotoxin were in the range of 26-6250 ng/m(3). In the samples of settled dust, the concentrations of total microorganisms ranged from 0.25 x 10(6) to 2.87 x 10(8) cfu/g. Gram-positive and Gram-negative bacteria constituted respectively 3.2-98 % and 0-93.5 % of the total count. Fungi formed 0-30.3 % of the total count. The most common species were Penicillium spp. and Alternaria alternata. The concentrations of endotoxin were in the range of 312.5-6250 microg/g (median 6250 microg/g). The presence of microorganisms and endotoxin in the samples of settled dust was confirmed by electron microscopy. The hop growers seem to be exposed to lower concentrations of dust, microorganisms and endotoxin compared to other branches of agriculture. This may be partly due to antimicrobial properties of hop plant. Among microbial factors associated with hop dust, bacterial endotoxin and allergenic fungi pose the greatest potential hazard for exposed hop farmers.

Aerosols↗
PubMedDate not supplied

Adaptation of the short latency component of the stretch reflex plays only a minor role in compensating for muscle fatigue induced by spontaneous hopping in humans.

The influence of fatigue on the stretch reflex evoked in ankle extensor muscles by hopping was investigated in six healthy men. The men hopped on a force platform, at spontaneous frequency and amplitude, until they were unable to maintain the initial frequency or amplitude of the jumps. This task was done with the knees flexing normally during ground contact or under instructions to straighten the knees. Surface electromyograms (EMG) of soleus (SO), gastrocnemius medialis (GM), and tibialis anterior (TA) muscles were recorded simultaneously with the vertical component of the ground reaction force. Spectrum analysis of the EMG recorded during isometric tests performed immediately before and after the fatiguing hopping task demonstrated the existence of fatigue in SO and GM and often in TA. The stretch reflex was studied during the first and last ten jumps of every hopping series. The long-latency components of the reflex were too variable to be analysed. Whatever the hopping condition, latency and amplitude of the short latency component were not significantly modified by fatigue. Fatigue enhanced the occurrence of this reflex component in SO only. These data suggest that in fatiguing submaximal hopping, the neuromuscular system does not fundamentally change its stiffness regulation before the endurance time has been reached.

Adaptation, Physiological↗
PubMed2003

STRO-1, HOP-26 (CD63), CD49a and SB-10 (CD166) as markers of primitive human marrow stromal cells and their more differentiated progeny: a comparative investigation in vitro.

There is widespread interest in the use of bone marrow stromal cells (BMSC) for tissue reconstruction and repair and for gene therapy. BMSC represent the differentiated progeny of CFU-F, which however comprise a developmentally heterogeneous population as is reflected in the cellular heterogeneity of the cell populations to which they give rise. We have compared the efficacy of monoclonal antibodies recognising a series of stromal antigens, viz. STRO-1, HOP-26, CD49a and SB-10/CD166, as tools for the enrichment of CFU-F prior to culture and as developmental markers for culture-expanded BMSC. In freshly isolated bone marrow mononuclear cells (BMMNC), the proportion of antigen-positive cells was 27%, 46%, 5% and 19% for STRO-1, HOP-26, CD49a and CD166, respectively. All CD49a(+) cells co-expressed STRO-1. The degree of CFU-F enrichment obtained with anti-CD49a (approximately 18-fold) by a one-pass immunoselection strategy was significantly greater than that of all other antibodies tested. BMSC expressed higher levels of all antigens investigated (except for HOP-26) compared with BMMNC. Expression of STRO-1 and CD49a remained restricted to a subset of BMSC, whereas all BMSC were SB-10/CD166 positive. Treatment with dexamethasone (10 nM), which promotes the differentiation and further maturation of cells of the osteogenic lineage in this cell culture system, increased the expression of CD49a and HOP-26. The CD49a(+) and HOP-26(+) fractions of BMSC were further subdivided by dual-labelling with anti-STRO-1 and B4-78 (an antibody recognising the B/L/K isoform of the enzyme alkaline phosphatase), respectively. By using a variety of criteria, the HOP-26 antigen was identified as CD63, a member of the tetraspanin family of proteins thought to modulate integrin compartmentalisation and signalling.

Activated-Leukocyte Cell Adhesion Molecule↗
PubMed2006

A first prospective, randomized, double-blind, placebo-controlled study on the use of a standardized hop extract to alleviate menopausal discomforts.

OBJECTIVES: To examine the efficacy of a hop extract enriched in 8-prenylnaringenin (8-PN, the phytoestrogen in hops, Humulus lupulus L.) on relief of menopausal discomforts. METHODS: A prospective, randomized, double-blind, placebo-controlled study over 12 weeks with 67 menopausal women, who were administered a hop extract standardized on 8-PN (100 or 250 microg). The responses were determined by means of a modified Kupperman index (KI) and a patients' questionnaire. RESULTS: All groups, including placebo, showed a significant reduction of the KI both after 6 weeks and after 12 weeks. The hop extract at 100 microg 8-PN was significantly superior to placebo after 6 weeks (P=0.023) but not after 12 weeks (P=0.086). No dose-response relationship could be established, as the higher dose (250 microg) was less active than the lower dose both after 6 weeks and after 12 weeks. Still, a trend for a more rapid decrease of KI was noticed for both active groups as compared to placebo. In particular, the decrease in hot flush score (isolated from the KI) was found significant for both treatment groups after 6 weeks (P<0.01) with respect to placebo. Results of the patients' questionnaire were consistent with those of the KI, with the most pronounced effects being observed for the 100-microg treatment. CONCLUSIONS: Daily intake of a hop extract, standardized on 8-PN as a potent phytoestrogen, exerted favorable effects on vasomotor symptoms and other menopausal discomforts. Hop-derived prenylated flavonoids may provide an attractive addition to the alternative treatments available for relief of hot flushes and other menopausal discomforts.

Double-Blind Method↗
PubMed2004

Electron hopping dynamics in monolayer-protected au cluster network polymer films by rotated disk electrode voltammetry.

Electrons are transported within polymeric films of alkanethiolate monolayer-protected Au clusters (MPCs) by electron hopping (self-exchange) between the metal cores. The surrounding monolayers, the molecular linkers that generate the network polymer film, or both, presumably serve as tunneling bridges in the electron transfers. This paper introduces a steady-state electrochemical method for measuring electron hopping rates in solvent-wetted and swollen, ionically conductive MPC films. The films are network polymer films of nanoparticles, coated on a rotated disk electrode that is contacted by a solution of a redox species (decamethylferrocene, CpFe). Controlling the electrode potential such that the film mediates oxidation of the redox probe can force control of the overall current onto the rate of electron hopping within the film, which is characterized as the apparent electron diffusion coefficient D(E). D(E) is translated into an apparent electron hopping rate k(ET) by a cubic lattice model. The experiment is applied to MPC network polymer films linked by alpha,omega-alkanedithiolates and by metal ion-carboxylate connections. We evaluate the dependencies of apparent hopping rate on CpFe concentration, film thickness, electrode potential relative to the CpFe formal potential, film-swelling solvent, and temperature. The apparent hopping rates are in the 10(4)-10(5) s(-)(1) range, which is slower than those for the same kind of MPC films, but in a dry (nonswollen) state measured by electronic conductivities.

Journal Article↗
PubMed2002

Method development and fate determination of pesticide-treated hops and their subsequent usage in the production of beer.

The fate of residues of seven agrochemicals (chlorfenapyr, quinoxyfen, tebuconazole, fenarimol, pyridaben, and E- and Z-dimethomorph) from the treatment on hops to the brewing of beer was studied. First, a multi-residue analytical method was developed for the determination of pesticide residues in spent hops, trub, wort, and beer. Each matrix was validated over at least two levels of fortification, for all seven compounds, in the ranges 0.05-5.0, 0.001-1.0, 0.001-0.05, and 0.0005-1.0 ppm for spent hops, trub, wort, and beer, respectively. Recoveries ranged from 73 to 136%. Second, the matrixes prepared from hops, which were treated under commercial practices with each compound, were analyzed using the method developed. The use of treated hops resulted in the carryover of 0.001 ppm of tebuconazole, 0.008 Z-dimethomorph, and 0.005 ppm of E-dimethomorph into the wort. The bulk of the remaining residues of all seven compounds was found on the spent hops. Following fermentation, all compounds were found in levels less than 0.0005 ppm in beer, except Z- (0.006 ppm) and E-dimethomorph (0.004 ppm). Third, when all seven pesticides were spiked prior to the pitching of yeast into clean wort, most of the nonpolar compounds (chlorfenapyr, quinoxyfen, and pyridaben) partitioned into the organic material (trub) which settled to the bottom, while the more polar compounds (fenarimol, tebuconazole, and E- and Z-dimethomorph) were generally distributed evenly between the beer and the trub.

Beer↗
PubMed2003

Formation and accumulation of alpha-acids, beta-acids, desmethylxanthohumol, and xanthohumol during flowering of hops (Humulus lupulus L.).

Important secondary metabolites, present in hops (Humulus lupulus L.), include alpha-acids and beta-acids, which are essential for the brewing of beer, as well as the prenylated chalcones, desmethylxanthohumol, and xanthohumol, which exhibit interesting bioactive properties. Their formation and accumulation in five selected hop varieties, Wye Challenger, Wye Target, Golding, Admiral, and Whitbread Golding Variety, were quantitatively monitored by high-performance liquid chromatography using UV detection. All target compounds were present from the onset of flowering, not only in female hop cones but also in male inflorescences, albeit in low concentrations. During development from female inflorescences to cones, levels of alpha-acids, beta-acids, desmethylxanthohumol, and xanthohumol gradually increased, while each hop variety exhibited individual accumulation rates. Furthermore, these compounds were present in leaves of fully grown hops as well. The study demonstrated that key compounds for flavor and potential beneficial health effects associated with beer not only reside in the glandular lupulin structures but also are distributed over various parts of the hop plant.

Beer↗
PubMed2005

Comparison of the in vitro estrogenic activities of compounds from hops (Humulus lupulus) and red clover (Trifolium pratense).

Because the prevailing form of hormone replacement therapy is associated with the development of cancer in breast and endometrial tissues, alternatives are needed for the management of menopausal symptoms. Formulations of Trifolium pratense L. (red clover) are being used to alleviate menopause-associated hot flashes but have shown mixed results in clinical trials. The strobiles of Humulus lupulusL. (hops) have been reported to contain the prenylflavanone, 8-prenylnaringenin (8-PN), as the most estrogenic constituent, and this was confirmed using an estrogen receptor ligand screening assay utilizing ultrafiltration mass spectrometry. Extracts of hops and red clover and their individual constituents including 8-PN, 6-prenylnaringenin (6-PN), isoxanthohumol (IX), and xanthohumol (XN) from hops and daidzein, formononetin, biochanin A, and genistein from red clover were compared using a variety of in vitro estrogenic assays. The IC50 values for the estrogen receptor alpha and beta binding assays were 15 and 27 microg/mL, respectively, for hops and 18.0 and 2.0 microg/mL, respectively, for the red clover extract. Both of the extracts, genistein, and 8-PN activated the estrogen response element (ERE) in Ishikawa cells while the extracts, biochanin A, genistein, and 8-PN, significantly induced ERE-luciferase expression in MCF-7 cells. Hop and red clover extracts as well as 8-PN up-regulated progesterone receptor (PR) mRNA in the Ishikawa cell line. In the MCF-7 cell line, PR mRNA was significantly up-regulated by the extracts, biochanin A, genistein, 8-PN, and IX. The two extracts had EC50 values of 1.1 and 1.9 microg/mL, respectively, in the alkaline phosphatase induction assay. On the basis of these data, hops and red clover could be attractive for the development as herbal dietary supplements to alleviate menopause-associated symptoms.

Alkaline Phosphatase↗
PubMed2000

Comparison of the most odor-active compounds in fresh and dried hop cones (Humulus lupulus L. variety spalter select) based on GC-olfactometry and odor dilution techniques.

Application of aroma extract dilution analysis on the volatiles obtained from dried cones of Spalter Select hops grown in the German hop-growing area of Hallertau revealed 23 odorants in the flavor dilution (FD) factor range of 16-4096, 20 of which could be identified. On the basis of high FD factors, trans-4, 5-epoxy-(E)-2-decenal, linalool, and myrcene were identified as the most potent odorants, followed by ethyl 2-methylpropanoate, methyl 2-methylbutanoate, (Z)-1,5-octadien-3-one, nonanal, (E,Z)-1,3, 5-undecatriene, 1,3(E),5(Z),9-undecatetraene, propyl 2-methylbutanoate, 4-ethenyl-2-methoxyphenol, and 1-octen-3-one. Ten of the high-impact hop aroma compounds had previously not been identified as hop constituents and, in particular, 1,3(E),5(Z), 9-undecatetraene has not yet been reported as a food odorant. In an extract obtained from fresh hops, in addition to the odorants found in dry hops, (Z)-3-hexenal was characterized as a further key odorant rendering an additional green aroma note to the fresh material.

Chromatography, Gas↗
PubMed2004

A simple method for field measurements of leg stiffness in hopping.

A new method to measure the leg stiffness in hopping and bouncing, with simple technical equipment and under field conditions, is introduced and validated. The leg stiffness (K (N)) was calculated from only contact and flight times measured by a contact mat. It was compared to the reference stiffness (K (R)) obtained from force platform measurements. Eight subjects performed, first, submaximal hopping movements at different frequencies (1.8 to 4 Hz, by step 0.2 Hz) and, second, maximal hopping. In sub maximal hopping K (N) was significantly correlated with K (R) (r = 0.94; p < 0.001) and the difference between K (N) and K (R) ranged from -7.2 % to 6.9 % (at 1.8 and 3.6 Hz respectively) with a limit of agreement of -1.5 kN x m (-1). In maximal hopping K (N) was also related to K (R) (r = 0.98, p < 0.001) and the inter individual rank order was respected (R = 0.87). It was concluded that the new method could be applied to study extensively intra individual and inter individual variations of leg stiffness in respectively sub maximal and maximal hopping and thus to simplify further investigations in field conditions of the role of stiffness regulation in the optimization of human locomotion.

Adult↗
PubMed2005

Entropy factor in the hopping frequency for ionic conduction in oxide glasses induced by energetic clustering.

The contribution of configurational entropy to the effective hopping frequency of ionic transport in amorphous systems is discussed. The effective rate of ion hopping has been extracted from the onset frequency of the ac conductivity measured in ionically conducting silicate glasses. Both the onset frequency and the dc conductivity exhibit Arrhenius-type thermal activation with similar values for the activation energy, DeltaEa=0.65+/-0.3 eV. The prefactor of the onset frequency results in nu0'=(1.05+/-0.05)x10(11) Hz, which is much lower than characteristic vibrational frequencies (10(13) Hz). Following standard hopping percolation theory, the long-range motion is dominated by a fraction of high-energy barriers that connect clusters of faster sites. The multiplicity of equivalent sites for ion hop entails a retardation of the effective jumping time with respect to the elementary hop. This effect can be assimilated into a negative activation entropy term in the frequency prefactor of the ion hopping rate, which depends on the features of energy clustering and accounts for the wide dispersion of nu0' reported for many conducting glasses. The model implies an effective percolation length of Lc approximately 7 nm, in good agreement with previous works.

Journal Article↗
PubMed1999

Reverse genetic analysis of Caenorhabditis elegans presenilins reveals redundant but unequal roles for sel-12 and hop-1 in Notch-pathway signaling.

Mutations in the human presenilin genes PS1 and PS2 cause early-onset Alzheimer's disease. Studies in Caenorhabditis elegans and in mice indicate that one function of presenilin genes is to facilitate Notch-pathway signaling. Notably, mutations in the C. elegans presenilin gene sel-12 reduce signaling through an activated version of the Notch receptor LIN-12. To investigate the function of a second C. elegans presenilin gene hop-1 and to examine possible genetic interactions between hop-1 and sel-12, we used a reverse genetic strategy to isolate deletion alleles of both loci. Animals bearing both hop-1 and sel-12 deletions displayed new phenotypes not observed in animals bearing either single deletion. These new phenotypes-germ-line proliferation defects, maternal-effect embryonic lethality, and somatic gonad defects-resemble those resulting from a reduction in signaling through the C. elegans Notch receptors GLP-1 and LIN-12. Thus SEL-12 and HOP-1 appear to function redundantly in promoting Notch-pathway signaling. Phenotypic analyses of hop-1 and sel-12 single and double mutant animals suggest that sel-12 provides more presenilin function than does hop-1.

Animals↗
PubMed2006

Efficient extraction of high-quality total RNA from various hop tissues (Humulus lupulus L.).

The isolation of high-quality RNA is a prerequisite for gene expression studies. RNA quality is of special relevance if PCR-based strategies such as cDNA-AFLP or RT-PCR are followed. Our molecular investigations of hop (Humulus lupulus L.) focus on genes that determine the biosynthesis of prenylflavonoids with interesting biological activities which accumulate in the lupulin glands. However, optimized protocols for RNA extraction from hop cones are not available. In this study, the RNeasy midi kit protocol was modified to isolate high amounts of total RNA from fresh and freeze-dried hop tissues, specifically leaves, female inflorescences, and lupulin-enriched hop cone fractions. The main difficulties in obtaining high RNA yields were related to specific features of hop, including the abundance of secondary metabolites and their accumulation in the sticky lupulin glands. The introduction of a number of modifications into the RNeasy midi kit protocol resulted in RNA of high quality, as assessed by spectrophotometry and electrophoresis in agarose gels. The protocol developed is currently being used to produce cDNA-AFLP fingerprints of hop cones and leaves for genetic screening. Furthermore, the method could provide RNA for further molecular studies, including Northern blot hybridizations and reverse transcription-polymerase chain reactions.

Blotting, Northern↗