The Effect of Growth Regulators on Quality Parameters and Ginsenosides Accumulation in Panax quinquefolium L. Roots

American ginseng (Panax quinquefolium L.) is a perennial medicinal herb originally grown in Canada and USA, and recently also in China, Australia, Holland and Poland. Several commercial preparations are produced from ginseng roots, that are known for their antifatigue, antitumor, antistress and immune system stimulating functions. The medicinal properties are due mainly to the active components – ginsenosides. In this work, the results of field cultivation experiments are presented that examine the effects of foliar application of several growth regulators on quality parameters and ginsenoside content of P. quinuefolium roots. The growth regulators tested, i.e., kinetin, daminozide, mixture of gibberellic acid (GA₃) with potassium salt of α-naphthalene acetic acid (kNAA) and new preparation – IPO-1 – benzimidazole derivative (obtained from the Institute of Organic Industry in Warsaw – at present during the process of patent), were applied at a concentration of 100 or 200 mg l⁻¹ in the middle of June in the 2nd year of vegetation. After 4 years of cultivation, the roots were dug up and dried, and subsequently the quantitative analysis of individual saponins (R_b1, R_b2, R_c, R_d, R_e, R_g1) by HPLC was performed. Growth regulators significantly affected quality parameters, morphological features and accumulation of individual and total ginsenosides in ginseng roots. Regardless of doses, the plant roots treated with growth regulators had a higher content of total ginsenosides in comparison to the control. The growth regulators also affected individual ginsenosides level and narrowed the ratio of R_b:R_g group. The application of kinetin, daminozide and benzimidazole derivative for foliar spray during 2nd year of American ginseng vegetation caused a significant increase in air dry weight of roots and aboveground parts whereas the mixture of GA₃ and kNAA showed a decreasing effect. An increase of roots size was observed using higher doses (200 mg l⁻¹) of kinetin and daminozide while a decreasing tendency appeared with the application of the other preparations.     

The Evolution of MHC Diversity by Segmental Duplication and Transposition of Retroelements

Sequence analysis of a 237 kb genomic fragment from the central region of the MHC has revealed that the HLA-B and HLA-C genes are contained within duplicated segments peri-B (53 kb) and peri-C (48 kb), respectively, and separated by an intervening sequence (IF) of 30 kb. The peri-B and peri-C segments share at least 90% sequence homology except when interrupted by insertions/deletions including Alu, L1, an endogenous retrovirus, and pseudogenes. The sequences of peri-B, IF, and peri-C were searched for the presence of Alu elements to use as markers of evolution, chromosomal rearrangements, and polymorphism. Of 29 Alu elements, 14 were identified in peri-B, 11 in peri-C, and 4 in IF. The Alu elements in peri-B and peri-C clustered phylogenetically into two clades which were classified as ``preduplication' and ``postduplication' clades. Four Alu J elements that are shared by peri-B and peri-C and are flanked by homologous sequences in their paralogous locations, respectively, clustered into a ``preduplication' clade. By contrast, the majority of Alu elements, which are unique to either peri-B or peri-C, clustered into a postduplication clade together with the Alu consensus subfamily members ranging from platyrrhine-specific (Spqxcg) to catarrhine-specific Alu sequences (Y). The insertion of platyrrhine-specific Alu elements in postduplication locations of peri-B and peri-C implies that these two segments are the products of a duplication which occurred in primates prior to the divergence of the New World primate from the human lineage (35–44 mya). Examination of the paralogous Alu integration sites revealed that 9 of 14 postduplication Alu sequences have produced microsatellites of different length and sequence within the Alu 3′-poly A tail. The present analysis supports the hypothesis that HLA-B and HLA-C genes are products of an extended segmental duplication between 44 and 81 million years ago (mya), and that subsequent diversification of both genomic segments occurred because of the mobility and mutation of retroelements such as Alu repeats. Received: 21 May 1997 / Accepted: 9 July 1997     

Light-induced isomerization of the LHCII-bound xanthophyll neoxanthin: possible implications for photoprotection in plants

Light-harvesting pigment-protein complex of Photosystem II (LHCII) is the largest photosynthetic antenna complex of plants and the most abundant membrane protein in the biosphere. Plant fitness and productivity depend directly on a balance between excitations in the photosynthetic apparatus, generated by captured light quanta, and the rate of photochemical processes. Excess excitation energy leads to oxidative damage of the photosynthetic apparatus and entire organism and therefore the balance between the excitation density and photosynthesis requires precise and efficient regulation, operating also at the level of antenna complexes. We show that illumination of the isolated LHCII leads to isomerization of the protein-bound neoxanthin from conformation 9'-cis to 9',13- and 9',13'-dicis forms. At the same time light-driven excitation quenching is observed, manifested by a decrease in chlorophyll a fluorescence intensity and shortened fluorescence lifetimes. Both processes, the neoxanthin isomerization and the chlorophyll excitation quenching, are reversible in dim light. The results of the 77K florescence measurements of LHCII show that illumination is associated with appearance of the low-energy states, which can serve as energy traps in the pigment-protein complex subjected to excess excitation. Possible sequence of the molecular events is proposed, leading to a protective excess excitation energy quenching: neoxanthin photo-isomerization→formation of LHCII supramolecular structures which potentiate creation of energy traps→excitation quenching.     

Early Cambrian lobopodian sclerites and associated fossils from Kazakhstan

A low diversity fossil assemblage dominated by Mongolitubulus spines, from the Early Cambrian Geres Member of the Shabakhty Formation at Koksu, Malyi Karatau, Kazakhstan, enables identification of its sclerite variability within the scleritome. No other sclerotised structures could be matched with the spines, indicating that these were the only sclerotised part of the body and were periodically shed. The prominent spines of Mongolitubulus have a scale–like external ornamentation and fibrous internal structure. Possibly they armed the body of a lobopodian similar to Xenusion . The Mongolitubulus spines belong to a series of Cambrian phosphatic fossils, the other end members of which are sclerites of Microdictyon , the second most abundant component of the Koksu assemblage. The latter superficially resemble schizochroal trilobite eyes, but in life covered the dorso–lateral sides of the body segments, as is evident from complete specimens from the Chinese Chengjiang locality. A pair of sclerites of Microdictyon , representing uncompleted exuviation, from Koksu show that each hole originally contained a non–phosphatised lenticular body. This makes a compound eye nature of the Microdictyon sclerites likely, and even their homology with arthropod eyes cannot be excluded.     

Antibiotic resistance of staphylococci isolated from outpatients

The aim of the study was to determine susceptibility of 587 strains of S. aureus and 85 strains of coagulase-negative staphylococci isolated from outpatients in Poznań to co-trimoxazole, amoxycillin/clavulanic acid, erythromycin, gentamycin, doxycycline, ampicillin, oxacillin, cephradine, clindamycin and neomycin. Also methicillin-resistant strains were determined as well as strains ability to produce beta-lactamases. Susceptibility testing and examination of methicillin-resistant strains were performed by the disc diffusion techniques according to recommendation of NCCLS. Methicillin-resistant strains were additionally examined to their sensitivity to vankomycin and teicoplanin. beta-lactamase production was detected using nitrocefin impregnated discs and iodometric method. Amoxacillin/clavulanic acid, gentamycin, co-trimoxazole, cephradin, oxacillin and clindamycin occurred to be very active against both, S. aureus and coagulase-negative staphylococci. 84.7% to 100% of examined strains were sensitive to these drugs. Doxycyclin, erythromycin and ampicillin were less effective. Nine strains (1.5%) of 587 strains of S. aureus as well as 7 strains (8.7%) of coagulase-negative staphylococci were methicillin-resistant. All of methicillin-resistant strains were sensitive to vancomycin and teicoplanin. More than 75% of S. aureus and close to 50% of coagulase-negative staphylococci were able to produce beta-lactamases.     

Ovary architecture of two branchiobdellid species and Acanthobdella peledina (Annelida, Clitellata)

The aim of this study was to present data about ovary organization and oogenesis in two small groups of clitellate annelids, i.e. in representatives of Acanthobdellida (Acanthobdella peledina) and Branchiobdellida (Branchiobdella pentodonta and Branchiobdella parasitica), and to compare them to ovaries known from true leeches and oligochaetous clitellates. In A. peledina, the ovaries have the form of elongated cords, termed ovary cords, and are enveloped by coelomic sacs, the so-called ovisacs. The ovisacs are paired and each one contains only one ovary cord. The morphology and structure of the ovary cords depend on the maturity level of the animal. In young specimens the ovary cords are short and contain mainly oogonial cells and germ cells entering meiosis. Oogonia divide mitotically without full cytokineses, and as a result germ-line cysts are formed. As the animals grow, the cords become more elongated and the germ cells within the cords differentiate into nurse cells and oocytes. Oocytes gather cell organelles and, finally, detach from the ovary cord and float freely in the ovisac lumen.In both examined branchiobdellidans the ovaries are also paired. They are short and conical and are not enclosed within ovisacs. The narrow end of each ovary is connected to the intersegmental septum via a ligament, whereas the outermost (broad) end of the ovary extends freely into the coelom. The ovaries are polarized. Their narrow ends contain oogonia, whereas nurse cells and growing oocytes, gradually projecting from the ovary, can be found in their middle and outermost parts. Early vitellogenic oocytes detach from the ovary and float freely in the coelom.In all of the species studied, the ovaries are made up of germ-line cysts associated with somatic (follicular) cells. The architecture of a germ-line cyst is exactly the same as in other clitellate annelids that have been studied to date. Each germ cell in a cyst has one stable cytoplasmic bridge connecting it with a central anuclear cytoplasmic mass, a cytophore. The fate of germ cells constituting cysts is diverse. The majority of the cells withdraw from meiosis and become nurse cells; only a few continue meiosis, grow and become oocytes. The meroistic mode of oogenesis is suggested. We suggest also that the formation of germ-line cysts and ovary meroism should be regarded as basal conditions for all Clitellata. The occurrence of ovisacs enveloping the ovaries in A. peledina and Hirudinida is regarded as a synapomorphy of both groups, whereas ovaries found in B. pentodonta and B. parasitica have no ovisacs and resemble ovaries described in Oligochaeta sensu stricto.     

Melatonin Increases Serotonin N-Acetyltransferase Activity and Decreases Dopamine Synthesis in Light-Exposed Chick Retina: In Vivo Evidence Supporting Melatonin-Dopamine Interaction in Retina

The administration of melatonin, either peripherally (0.01-10 mg/kg) or intraocularly (0.001-10 mumol/eye), to light-exposed chicks dose-dependently increased serotonin N-acetyltransferase (NAT) activity in retina but not in pineal gland. The effect of melatonin was slightly but significantly reduced by luzindole (2-benzyl-N-acetyltryptamine), and not affected by two other purported melatonin antagonists, N-acetyltryptamine and N-(2,4-dinitrophenyl)-5-methoxytryptamine (ML-23). The elevation of the enzyme activity induced by melatonin was substantially stronger than that evoked by 5-hydroxytryptamine, N-acetyl-5-hydroxytryptamine, or 5-methoxytryptamine. The melatonin-evoked rise in the retinal NAT activity was counteracted by two dopamine D2 receptor agonists, quinpirole and apomorphine, and prevented by the dopamine D2 receptor blocker spiroperidol, and by an inhibitor of dopamine synthesis, alpha-methyl-p-tyrosine. Melatonin (0.1-10 mg/kg i.p.) dose-dependently decreased the levels of dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC), as well as the DOPAC/dopamine ratio, in chick retina but not in forebrain. The results obtained (1) indicate that melatonin in vivo potently inhibits dopamine synthesis selectively in retina, and (2) suggest that the increase in retinal NAT activity evoked by melatonin in light-exposed chicks is an indirect action of the compound, and results from the disinhibition of the NAT induction process from the dopaminergic (inhibitory) signal. The results provide in vivo evidence supporting the idea (derived on the basis of in vitro findings) that a mutually antagonistic interaction between melatonin and dopamine operates in retinas of living animals.