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Identification of an antibacterial compound, benzylideneacetone, from Xenorhabdus nematophila against major plant-pathogenic bacteria.

An entomopathogenic bacterium, Xenorhabdus nematophila, is known to have potent antibiotic activities to maintain monoxenic condition in its insect host for effective pathogenesis and ultimately for optimal development of its nematode symbiont, Steinernema carpocapsae. In this study we assess its antibacterial activity against plant-pathogenic bacteria and identify its unknown antibiotics. The bacterial culture broth had significant antibacterial activity that increased with development of the bacteria and reached its maximum at the stationary growth phase. The antibiotic activities were significant against five plant-pathogenic bacterial strains: Agrobacterium vitis, Pectobacterium carotovorum subsp. atrosepticum, P. carotovorum subsp. carotovorum, Pseudomonas syringae pv. tabaci, and Ralstonia solanacearum. The antibacterial factors were extracted with butanol and fractionated using column chromatography with the eluents of different hydrophobic intensities. Two active antibacterial subfractions were purified, and the higher active fraction was further fractionated and identified as a single compound of benzylideneacetone (trans-4-phenyl-3-buten-2-one). With heat stability, the synthetic compound showed equivalent antibiotic activity and spectrum to the purified compound. This study reports a new antibiotic compound synthesized by X. nematophila, which is a monoterpenoid compound and active against some Gram-negative bacteria.

Animals↗

Expression of transgenic stilbene synthases in wheat causes the accumulation of unknown stilbene derivatives with antifungal activity.

The expression of foreign phytoalexins in a new host is thought to increase fungal resistance, since host-specific pathogens have not experienced selection for detoxifying or metabolising the novel antifungal compounds. Two resveratrol synthase genes vst1 and vst2 from grapevine (Vitis vinifera L.) and the pinosylvin synthase gene pss from pine (Pinus sylvestris L.) were stably transformed into bread wheat. The expression of the target genes is regulated by stress-inducible grapevine promoters. The vst1 and vst2 promoters were functional in wheat and retained their expression profiles described for grapevine. ALL vst and pss transgenic lines accumulated stilbene derivatives upon induction by UV light. The detected stilbenes showed a remarkable similarity to resveratrol and pinosylvin, however were found to be more hydrophilic than resveratrol and pinosylvin. Upon inoculation with the biotrophic pathogen Puccinia recondita f.sp. tritici several vst expressing wheat lines showed a significant reduction of disease symptoms (19 +/- 9% to 27 +/- 8%) compared to wild-type plants. The reduction of disease symptoms was even more obvious after inoculation with the facultative biotrophic pathogen Septoria nodorum Berk. and ranged from 42 +/- 13% to 71 +/- 4%. None of the four tested pss expressing lines showed a reduction in disease incidence.

Acyltransferases↗

RT-PCR-RFLP for genetic diversity analysis of Tunisian Grapevine fanleaf virus isolates in their natural host plants.

Genetic diversity was characterized in 20 isolates of Grapevine fanleaf virus (GFLV) recovered from naturally infected grapevine plants (Vitis vinifera) in the North of Tunisia. Viral RNAs were isolated by oligoprobe capture, and a 605 bp fragment containing a part of the viral coat protein gene was amplified by RT-PCR. Sequence variation among isolates was characterized by restriction fragment length polymorphism (RFLP) analysis and confirmed by sequencing. The GFLV infections are found as a complex mixture of closely related genomes. In further studies, RFLP analyses of virus isolates using AluI showed that GFLV populations in Tunisian vineyards consist of two restrictotypes corresponding to distinct sub-populations Sp1 and Sp2. The relative field distribution of these sub-populations showed that Sp2 was more abundant. Individual genomes were recovered by cloning the RT-PCR products. The sequences were found to vary from each other by as much as 11%. Cloning from mixed infections showed that Sp2 are also predominant.

Genetic Variation↗

Understanding local Mediterranean diets: a multidisciplinary pharmacological and ethnobotanical approach.

Epidemiological data indicate a beneficial effect of Mediterranean diets on human health, especially on the prevalence of cardiovascular disease. These observations are supported by recent intervention studies. However, very little is known about the current role of local Mediterranean food products, which are consumed on a less regular basis and their contribution to a healthy diet. The European consortium "Local Food-Nutraceuticals" collected 127 locally consumed wild or semi-wild plants in three Mediterranean countries, i.e. Greece, Italy, and Spain, in order to assess their ethnobotanical features as well as their biological activities. The project also includes a second line of research, the study of local conceptions about these food resources. All pharmacological assays were conducted with ethanolic extracts prepared from the dried plant material. The biological activities of the extracts were assessed with the following 12 different assays covering a broad range of mechanisms considered crucial in the pathology of chronic, aging-related diseases. Four antioxidant tests: DPPH scavenging, prevention of oxyhaemoglobin bleaching, prevention of lipid peroxidation (malondialdehyde formation), and protection from DNA damage (Comet assay); three enzyme inhibition tests: inhibition of xanthine oxidase, inhibition of myeloperoxidase-catalysed guaiacol oxidation as well as the inhibition of acetylcholine esterase; one test investigating the inhibition of cytokine-induced cell activation (including the extracts' potential cytotoxicity); one assay measuring the anti-proliferation potential; one test assessing the anti-diabetic activity (PPARgamma) as well as one assay investigating the extracts' effect on mood disorder-related biochemical parameters (hSERT). Furthermore, the polyphenol content of all extracts was determined using the Folin-Ciocalteaus method. The assays revealed diverse biological effects for the tested extracts ranging from no activity to almost complete inhibition/activation. Moreover, the experimental matrix led to the identification of a sub-set of extracts, i.e. Berberis vulgaris, Reichardia picroides, Scandix australis, Satureja montana, Thymus piperella, Lythrum salicaria and Vitis vinifera, showing high activity in a broad range of assays. In summary, the in vitro observed modulations and effects exerted by extracts derived from local food plants suggest that these plants may contribute to the observed better aging of rural Mediterranean populations.

Acetylcholinesterase↗

Diet enriched with procyanidins enhances antioxidant activity and reduces myocardial post-ischaemic damage in rats.

Aim of this work was to study the efficacy of procyanidins from Vitis vinifera seeds, a standardized mixture of polyphenol antioxidants, on cardiac mechanics following ischemia/reperfusion stunning in the rat, after 3 weeks supplementation. Young and aged male rats were fed a diet enriched with procyanidins complexed (1:3 w/w) with soybean lecithin (2.4%); control animals (CTR-young and CTR-aged) received an equal amount of lecithin and 2 additional groups of animals the standard diet. At the end of the treatment, the total plasma antioxidant defense (TRAP), vitamin E, ascorbic acid and uric acid were determined in plasma and the hearts from all groups of animals subjected to moderate ischemia (flow reduction to 1 ml/min for 20 min) and reperfusion (15 ml/min for 30 min). In both young and aged rats supplemented with procyanidins the recovery of left ventricular developed pressure (LVDP) at the end of reperfusion was 93% (p < 0.01) and 74% (p < 0.01) of the preischemic values and the values of coronary perfusion pressure (CPP) were maintained close to those of the preischemic period. Also creatine kinase (CK) outflow was restrained to baseline levels, while a 2-fold increase in prostacyclin (6-keto-PGF1alpha) in the perfusate from hearts of young and aged rats was elicited during both ischemia and reperfusion. In parallel, procyanidins significantly increased the total antioxidant plasma capacity (by 40% in young and by 30% in aged rats) and the plasma levels of ascorbic acid, while tend to reduce vitamin E levels; no significant differences were observed in uric acid levels. The results of this study demonstrate that procyanidins supplementation in the rat (young and aged) makes the heart less susceptible to ischemia/reperfusion damage and that this is positively associated to an increase in plasma antioxidant activity.

Aging↗

Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: down-regulation of COX-2 and iNOS through suppression of NF-kappa B activation.

A wide array of phenolic substances, particularly those present in edible and medicinal plants, have been reported to possess substantial anticarcinogenic and antimutagenic activities. The majority of naturally occurring phenolics retain antioxidative and anti-inflammatory properties which appear to contribute to their chemopreventive or chemoprotective activity. Cyclooxygenase-2 (COX-2) inducible and nitric oxide synthase (iNOS) are important enzymes that mediate inflammatory processes. Improper up-regulation of COX-2 and/or iNOS has been associated with pathophysiology of certain types of human cancers as well as inflammatory disorders. Since inflammation is closely linked to tumor promotion, substances with potent anti-inflammatory activities are anticipated to exert chemopreventive effects on carcinogenesis, particularly in the promotion stage. Examples are curcumin, a yellow pigment of turmeric (Curcuma longa L., Zingiberaceae), the green tea polyphenol epigallocatechin gallate (EGCG), and resveratrol from grapes (Vitis vinifera, Vitaceae) that strongly suppress tumor promotion. Recent studies have demonstrated that eukaryotic transcription factor nuclear factor-kappa B (NF-kappa B) is involved in regulation of COX-2 and iNOS expression. Several chemopreventive phytochemicals have been shown to inhibit COX-2 and iNOS expression by blocking improper NF-kappa B activation. Multiple lines of compelling evidence indicate that extracellular-regulated protein kinase and p38 mitogen-activated protein kinase are key elements of the intracellular signaling cascades responsible for NF-kappa B activation in response to a wide array of external stimuli. Curcumin, EGCG and resveratrol have been shown to suppress activation of NF-kappa B. One of the plausible mechanisms underlying inhibition of NF-kappa B activation by aforementioned phytochemicals involves repression of degradation of the inhibitory unit I kappa B alpha, which hampers subsequent nuclear translocation of the functionally active subunit of NF-kappa B.

Animals↗

Changes in grape seed polyphenols during fruit ripening.

The quantity and characterization of extracted flavan-3-ol monomers and procyanidins was determined in seeds from Vitis vinifera cv. Cabernet Sauvignon berries, over the course of ripening and at different levels of vine water status. The per berry extractive yield of all polyphenols decreased with maturity, and followed second-order kinetics. The flavan-3-ol monomers decreased most rapidly, followed by the procyanidin extension units and finally, the terminal units. The relative proportion of procyanidin extension units did not vary with maturity. During fruit ripening, the mean degree of polymerization of extracted procyanidins is unchanged when analyzed intact by HPLC, but decreases by thiolytic degradation. The proportion of extracted procyanidins resistant to acid catalyzed thiolysis increased with maturity. Changes in vine water status affected polyphenol amounts, indicating that cultural practices can be used to influence composition. Oxidation of the seed polyphenols during fruit ripening, could explain these observations.

Flavonoids↗

Specific transfer of 3H from D-[3-3H]gluconic acid into L-tartaric acid in vitaceous plants.

Transfer of 3H from D-gluconic acid, specifically labelled with 3H at C-2 or C-3 and 14C at C-1, C-2, or C-3, 4, to L(+)-tartaric acid was examined in leaves and berries of Vitis labrusca cv Delaware and in leaves of Parthenocissus quinquefolia. 3H located at C-3 of D-gluconic acid was highly conserved in this transfer, yielding a 3H/14C ratio between 3.3 and 14 in the light and between 11 and 22 in the dark. These experiments strongly suggest that a portion of the 3H present in L(+)-tartaric acid may have been transferred from D-gluconic acid to L(+)-tartaric acid, possibly via NADP[3H] through a redox process involving reduction of L-xylo-2-hexulosonate (2-keto-L-idonate). Both [3H]-tartaric acid and [14C]tartaric acid synthesized in grape leaves from D-[3-3H, 2-14C]gluconic acid, or [3-3H, 3,4-14C]gluconic acid were characterized as L(+)-chiral form exclusively, the naturally occurring from of tartaric acid.

Fruit↗

Effect of industrial pollution on the distribution of 137Cs in soil and the soil-to-plant transfer in a pine forest in SW Finland.

The objective of this study was to evaluate the effect of industrial pollution on the distribution of radiocaesium in soil and on its transfer from soil to plants. The study was started in September 2000 in four Scots pine stands located at distances of 0.5, 2, 4 and 8 km along a transect running SE from the Cu-Ni smelter at Harjavalta in SW Finland. Annual emissions from the smelter in 1990 were 80 t of Cu, 31 t of Ni and 9000 t of SO(2), and in 1999 these were 5.9, 0.8 and 3400 t, respectively. At each site, soil profiles were sampled with a corer, and samples were separated into litter (L), organic soil layer (O) and mineral soil layers (B, E). Mushrooms, lichens (Cladina spp. and Cetraria islandica), lingonberry (Vaccinium vitis-idaea) and crowberry (Empetrum nigrum) plants were collected at each site, except at a distance of 0.5 km, where only mushrooms were available. In the organic soil layer, 137Cs activity decreased from 8000 Bq/m(2) at a distance of 8 km from the smelter to 1500 Bq/m(2) at a distance of 0.5 km; in litter, 137Cs concentration increased from 6300 Bq/m(2) at 8 km to 14000 Bq/m(2) at 0.5 km. 137Cs activity concentration decreased significantly in plants, mushrooms and lichens as the pollution load increased. In lichens, 137Cs activity decreased from 910 Bq/kg at 8 km to 170 Bq/kg at 2 km, while in lingonberry it decreased from 1470 to 20 Bq/kg and in crowberry from 310 to 20 Bq/kg. Aggregated transfer factors for 137Cs decreased in a similar way in lingonberry from 7.6x10(-2) m(2)/kg at 8 km to 7.7x10(-4) m(2)/kg at 2 km and in crowberry from 1.6x10(-2) to 7.9x10(-4) m(2)/kg.

Agaricales↗

Rhinoconjunctivitis and asthma caused by vine pollen: a case report.

BACKGROUND: The vine (Vitis vinifera) is a cultivated plant that is found in some European and American countries. Its pollen gathers in small quantities during a short pollination period in the months of May and June. Allergy to vine pollen has not been previously documented. OBJECTIVE: We sought to describe a case report of allergy to vine pollen documented on the basis of anamnesis, cutaneous, provocation, and specific IgE determination tests. METHODS: An allergenic extract was obtained from collected V vinifera pollens by aqueous standard procedures. Pollen counts and pollination periods of this and other common pollens in the area where the patient became symptomatic were studied. Cutaneous tests and the presence of specific IgE to the pollen extracts were performed by prick, CAP, and RAST techniques. Bronchial and conjunctival tests with the involved pollen extracts were also carried out to identify the sensitizing allergens. Five healthy subjects and 5 pollinic patients were used as control subjects and underwent the same tests. RESULTS: Skin prick test responses with vine pollen at different concentrations were positive for the studied patient and negative for the control subjects. Patient serum revealed a total IgE titer of 334 IU/mL and a specific IgE value of 1.3 PRU/mL (RAST class 2) to vine pollen. Bronchial and conjunctival provocation test responses were also positive when the patient was challenged with V vinifera extract. CONCLUSION: Exposure to the pollen of the vineyard plants (V vinifera) can induce immunologic sensitization and rhinoconjunctivitis/asthma.

Adolescent↗

Influence of extreme pollution on the inorganic chemical composition of some plants.

Leaves of nine different plant species (terrestrial moss: Hylocomium splendens and Pleurozium schreberi, blueberry: Vaccinium myrtillus, cowberry: Vaccinium vitis-idaea, crowberry: Empetrum nigrum, birch: Betula pubescens, willow: Salix spp., pine: Pinus sylvestris, and spruce: Picea abies) have been collected from up to nine catchments (size 14-50 km2) spread over a 1,500,000 km2 area in northern Europe. Additional soil samples were taken from the O-horizon and the C-horizon at each plant sample site. All samples were analysed for 38 elements (Ag, Al, As, B, Ba, Be, Bi, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Li, Mg, Mn, Mo, Na, Ni, P, Pb, Rb, S, Sb, Sc, Se, Si, Sn, Sr, Th, Tl, U, V, Y, Zn, and Zr) by ICP-MS, ICP-AES or CV-AAS (Hg) techniques. One of the 9 catchments was located directly adjacent (5-10 km S) to the nickel smelter and refinery at Monchegorsk, Kola Peninsula, Russia. The high levels of pollution at this site are reflected in the chemical composition of all plant leaves. However, it appears that each plant enriches (or excludes) different elements. Elements emitted at trace levels, such as Ag, As and Bi, are relatively much more enriched in most plants than the major pollutants Ni, Cu and Co. The very high levels of SO2 emissions are generally not reflected by increases in plant total S-content. Several important macro-(P) and micro-nutrients (Mn, Mg, and Zn) are depleted in most plant leaves collected near Monchegorsk.

Air Pollutants↗

Antioxidant and eicosanoid enzyme inhibition properties of pomegranate seed oil and fermented juice flavonoids.

The antioxidant and eicosanoid enzyme inhibition properties of pomegranate (Punica granatum) fermented juice and seed oil flavonoids were studied. The pomegranate fermented juice (pfj) and cold pressed seed oil (pcpso) showed strong antioxidant activity close to that of butylated hydroxyanisole (BHA) and green tea (Thea sinensis), and significantly greater than that of red wine (Vitis vitifera). Flavonoids extracted from pcpso showed 31-44% inhibition of sheep cyclooxygenase and 69-81% inhibition of soybean lipoxygenase. Flavonoids extracted from pfj showed 21-30% inhibition of soybean lipoxygenase though no significant inhibition of sheep cyclooxygenase. The pcpso was analyzed for its polyphenol content and fatty acid composition. Total polyphenols in pcpso showed a concentration by weight of approximately 0.015%. Pcpso fatty acid composition showed punicic acid (65.3%) along with palmitic acid (4.8%), stearic acid (2.3%), oleic acid (6.3%), linoleic acid (6.6%) and three unidentified peaks from which two (14.2%) are probably isomers of punicic acid (El-Shaarawy, M.I., Nahpetian, A., 1983). Studies on pomegranate seed oil. Fette Seifen Anstrichmittel 83(3), 123-126).

Animals↗

Occurrence of resveratrol and pterostilbene in age-old darakchasava, an ayurvedic medicine from India.

'Darakchasava' is a well known Indian herbal preparation of which the main ingredient is Vitis vinifera L. This 'ayurvedic' medicine is prescribed as a cardiotonic and also given for other disorders. HPLC analysis of this age old formulation revealed the presence of polyphenols like resveratrol and pterostilbene. These phenolic compounds are now known as antioxidants, cancer chemopreventive agents, and also known to reduce mortality from coronary heart disease by increasing high density lipoproteins like cholesterol and inhibiting platelet aggregation (Soleas, J.S., Diamandis, E.P., Goldberg, D.M., 1997. Resveratrol: a molecule whose time has come? and gone? Clin. Biochem. 30 (2), 91-113). The study of darakchasava becomes interesting in the light of these findings. A brief introduction of this medicinal preparation, its formulation, its analysis by HPLC, and some of its properties are discussed in this article.

Antineoplastic Agents, Phytogenic↗

Phylogenetic analysis of 23S rRNA gene sequences of. Agrobacterium, Rhizobium and Sinorhizobium strains.

The phylogenetic relationship among twelve Agrobacterium, four Rhizobium, and two Sinorhizobium strains originating from various host plants and geographical regions was studied by analysis of the 23S rDNA sequences. The study included Agrobacterium strains belonging to biovars 1, 2 (with tumor- or hairy-root inducing and non-pathogenic strains), A. vitis, A. rubi; representative species of the Rhizobium genus: R. galegae, R. leguminosarum and R. tropici and Sinorhizobium meliloti strains. The phylogenetic analysis showed that within Agrobacterium, the biovar designation was reflected in the 23S rDNA similarity and that strains of Agrobacterium and Rhizobium are closely related to each other. The results suggest that the taxonomic definition of Agrobacterium and Rhizobium should be considered for revision and that the Agrobacterium-biovar identity is probably a reliable taxonomic trait.

Bacterial Typing Techniques↗

Free radical scavenging action of medicinal herbs from Mongolia.

In the present study we evaluated the free radical scavenging action of some medicinal herbs growing in Mongolia. The aqueous extract of nine herbs Chamenerion angustifolium (Ch.ang), Equisetum arvense (Eq.arv), Gentiana decumbens (Gn.dec), Geranium pratense (Gr.pra), Lomatogonium carinthiacum (L.car), Nonea poulla (N.pl), Phodococcum vitis-idaea (Ph.v), Sphallerocarpus gracilis (Sph.gr), Stellera chamaejasme (St.cha) were used in the present experiment. The free radical scavenging action was determined in vitro and ex vivo by using electron spin resonance (ESR) spectrometer and chemiluminescence (CL) analyzer. The results showed that extracts of Ch.ang, Gn.dec, Gr.pra, L.car, N.pl, Ph.v, Sph.gr and St. cha possess strong scavenging action of 1,1-diphenyl-2-picrylhydrazyl, superoxide and hydroxyl radicals. On the other hand, the radical scavenging action of Eq.arv was low. Extracts of N.pl and Ph.v markedly inhibited the CL generated from rat liver microsomal cytochrome P450 system whereas the CL was moderately inhibited by Eq.arv, Gn.dec, Gr.pra, L.car and St.cha. The extracts of Ch.ang and Sph.gr did not decrease the CL generation. Ch.ang, Gr.pra, L.car, N.pl, Ph.v and St.cha also depressed reactive oxygen production from polymorphonuclear leukocytes stimulated by phorbol-12-myristate acetate ex vivo. Thus it was confirmed that the medicinal herbs from Mongolia possess high antioxidant potency in vitro and ex vivo.

Animals↗

Adult onset grape hypersensitivity causing life threatening anaphylaxis.

BACKGROUND: Adverse reactions to foods are encountered much less frequently in adults than in the children. Adult onset hypersensitivity to grapes has not been previously reported. OBJECTIVE: Evaluation of a case of anaphylaxis that occurred as a result of the consumption of white grapes (Vitis vinifera). METHODS AND RESULTS: A 28-year-old woman experienced generalized urticaria, facial/oropharyngeal angioedema, and dizziness after eating a bunch of white grapes. She was treated in an emergency room for anaphylaxis. Previously, she had experienced two similar episodes after eating white grapes. The grape prick skin tests were strongly positive forming a pseudopod type reaction. The total serum IgE was 1918 ng/mL. The grape-specific serum IgE was weakly positive by the modified RAST and negative in the Pharmacia-Upjohn Cap System. CONCLUSION: Hypersensitivity to a commonly consumed fruit such as grapes can develop late in life causing a near-fatal anaphylaxis.

Adult↗

Metabolic engineering of Saccharomyces cerevisiae for the synthesis of the wine-related antioxidant resveratrol.

The stilbene resveratrol is a stress metabolite produced by Vitis vinifera grapevines during fungal infection, wounding or UV radiation. Resveratrol is synthesised particularly in the skins of grape berries and only trace amounts are present in the fruit flesh. Red wine contains a much higher resveratrol concentration than white wine, due to skin contact during fermentation. Apart from its antifungal characteristics, resveratrol has also been shown to have cancer chemopreventive activity and to reduce the risk of coronary heart disease. It acts as an antioxidant and anti-mutagen and has the ability to induce specific enzymes that metabolise carcinogenic substances. The objective of this pilot study was to investigate the feasibility of developing wine yeasts with the ability to produce resveratrol during fermentation in both red and white wines, thereby increasing the wholesomeness of the product. To achieve this goal, the phenylpropanoid pathway in Saccharomyces cerevisiae would have to be introduced to produce p-coumaroyl-CoA, one of the substrates required for resveratrol synthesis. The other substrate for resveratrol synthase, malonyl-CoA, is already found in yeast and is involved in de novo fatty-acid biosynthesis. We hypothesised that production of p-coumaroyl-CoA and resveratrol can be achieved by co-expressing the coenzyme-A ligase-encoding gene (4CL216) from a hybrid poplar and the grapevine resveratrol synthase gene (vst1) in laboratory strains of S. cerevisiae. This yeast has the ability to metabolise p-coumaric acid, a substance already present in grape must. This compound was therefore added to the synthetic media used for the growth of laboratory cultures. Transformants expressing both the 4CL216 and vst1 genes were obtained and tested for production of resveratrol. Following beta-glucosidase treatment of organic extracts for removal of glucose moieties that are typically bound to resveratrol, the results showed that the yeast transformants had produced the resveratrol beta-glucoside, piceid. This is the first report of the reconstruction of a biochemical pathway in a heterologous host to produce resveratrol.

Acyltransferases↗

A study of immunity to rubella in villages in the Fiji islands using the haemagglutination inhibition test.

In the villages of Fiji, apart from Viti Levu, rubella is a disease occurring solely in widely spaced epidemics. Some villages may not be infected for over 20 years and will then contain substantial numbers of susceptible women of child-bearing age.Evidence is produced that haemagglutination-inhibiting (H.I.) antibody to rubella is very long lasting in Fijians. The infectivity of the virus is discussed and it is suggested that, on the average, 50% of susceptibles are infected in a Fijian village during a rubella epidemic, but there are large variations.

Adolescent↗