Hantavirus-Leptospira coinfections in small mammals from central Germany.

European orthohantaviruses (Puumala orthohantavirus (PUUV); Dobrava-Belgrade orthohantavirus (DOBV), genotype Kurkino; Tula orthohantavirus (TULV)), and Leptospira spp. are small mammal-associated zoonotic pathogens that cause diseases with potentially similar symptoms in humans. We investigated the frequency of Leptospira spp. and hantavirus single and double infections in small mammals from 22 sites in Thuringia, central Germany, during 2017. TULV infections were detected at 18 of 22 sites (mean prevalence 13.8%, 93/674). PUUV infections were detected at four of 22 sites (mean prevalence 1.5%, 7/471), and respective PUUV sequences formed a novel phylogenetic clade, but DOBV infections were not detected at all. Leptospira infections were detected at 21 of 22 sites with the highest overall prevalence in field voles (Microtus agrestis) with 54.5% (6/11) and common voles (Microtus arvalis) with 30.3% (205/676). Leptospira-hantavirus coinfections were found in 6.6% (44/671) of common voles but only in two of 395 bank voles. TULV and Leptospira coinfection probability in common voles was driven by individual (age) and population-level factors. Coinfections seemed to be particularly associated with sites where Leptospira spp. prevalence exceeded 35%. Future investigations should evaluate public health consequences of this strong spatial clustering of coinfections.

Reservoir-Driven Heterogeneous Distribution of Recorded Human Puumala virus Cases in South-West Germany.

Endemic regions for Puumala virus (PUUV) are located in the most affected federal state Baden-Wuerttemberg, South-West Germany, where high numbers of notified human hantavirus disease cases have been occurring for a long time. The distribution of human cases in Baden-Wuerttemberg is, however, heterogeneous, with a high number of cases recorded during 2012 in four districts (H districts) but a low number or even no cases recorded in four other districts (L districts). Bank vole monitoring during 2012, following a beech (Fagus sylvatica) mast year, resulted in the trapping of 499 bank voles, the host of PUUV. Analyses indicated PUUV prevalences of 7-50% (serological) and 1.8-27.5% (molecular) in seven of eight districts, but an absence of PUUV in one L district. The PUUV prevalence differed significantly between bank voles in H and L districts. In the following year 2013, 161 bank voles were trapped, with reduced bank vole abundance in almost all investigated districts except one. In 2013, no PUUV infections were detected in voles from seven of eight districts. In conclusion, the linear modelling approach indicated that the heterogeneous distribution of human PUUV cases in South-West Germany was caused by different factors including the abundance of PUUV RNA-positive bank voles, as well as by the interaction of beech mast and the proportional coverage of beech and oak (Quercus spec.) forest per district. These results can aid developing local public health risk management measures and early warning models.

Validation of the Puumala virus rapid field test for bank voles in Germany.

Puumala virus (PUUV) causes many human infections in large parts of Europe and can lead to mild to moderate disease. The bank vole (Myodes glareolus) is the only reservoir of PUUV in Central Europe. A commercial PUUV rapid field test for rodents was validated for bank-vole blood samples collected in two PUUV-endemic regions in Germany (North Rhine-Westphalia and Baden-Württemberg). A comparison of the results of the rapid field test and standard ELISAs indicated a test efficacy of 93-95%, largely independent of the origin of the antigens used in the ELISA. In ELISAs, reactivity for the German PUUV strain was higher compared to the Swedish strain but not compared to the Finnish strain, which was used for the rapid field test. In conclusion, the use of the rapid field test can facilitate short-term estimation of PUUV seroprevalence in bank-vole populations in Germany and can aid in assessing human PUUV infection risk.

Environmental conditions in favour of a hantavirus outbreak in 2015 in Germany?

Bank voles can harbour Puumala virus (PUUV) and vole populations usually peak in years after beech mast. A beech mast occurred in 2014 and a predictive model indicates high vole abundance in 2015. This pattern is similar to the years 2009/2011 when beech mast occurred, bank voles multiplied and human PUUV infections increased a year later. Given similar environmental conditions in 2014/2015, increased risk of human PUUV infections in 2015 is likely. Risk management measures are recommended.

The effect of Kraussianone-2 (Kr2), a natural pyrano-isoflavone from Eriosema kraussianum, in an L-NAME- induced pre-eclamptic rat model.

This study aimed to investigate the effects of Kraussianone-2 (Kr2), a pyrano-isoflavone isolated from the roots of Eriosema kraussianum N. E. Br. (Fabaceae) on various fetal and physiological parameters in pregnant, L-NAME treated Sprague-Dawley rats. Twenty-four pregnant Sprague-Dawley dams were divided into three groups (n = 8), i.e. the control group (CON), the experimental control group (PRE), where the pre-eclampsia-like symptoms were induced using L-NAME, and the experimental group (EK2), where the pre-eclampsia-like symptoms were once again induced using L-NAME, however, these animals were treated with Kr2. On gestation day 20 the animals were sacrificed, at which time a laparotomy was performed and the number of live pups were counted and their corresponding birth and placental weights were recorded. Blood was also collected in heparin-coated tubes and the plasma samples were then analysed for specific variables using commercially available kits for rats. Kraussianone-2 administration decreased fetal mortality and demonstrated a trend toward increasing birth and placental weights in this model. Furthermore, Kr2 administration also reduced blood pressure amplification and decreased the plasma concentrations of two antiangiogenic factors, soluble fms-like tyrosine kinase1 (sFlt-1) and soluble endoglin (sEng). We speculate that Kr2, by improving uterine artery blood flow, results in improved fetal outcomes and decreased antiangiogenic factors in pregnant, L-NAME treated, Sprague-Dawley rats.

Hypoxis hemerocallidea--not merely a cure for benign prostate hyperplasia.

The use of Hypoxis hemerocallidea Fisch. and C.A. Mey. (Hypoxidaceae) extracts for ailments other than benign prostate hyperplasia (BPH) is now a reality. The whole extract, the phytosterols, as well as the major constituents it contains (hypoxoside, and its active derivative rooperol) are now finding new applications in the area of anti-oxidants, anti-inflammatories, anti-diabetics, anti-convulsants, inhibitors of drug marker substances and new evidence is presented of activity against cancerous and premalignant cancer cells. In addition, the over-the-counter (OTC) trade has undergone enormous expansion.

Transformation of naturally-occurring 1,9-trans-9,5-cis sweroside to all trans sweroside during acetylation of sweroside aglycone.

From the rootstock of Scabiosa columbria L. loganin and sweroside were isolated. Sweroside showed moderate antibacterial activity. Pure sweroside was converted to the sweroside aglycone 1-acetoxy derivative (DABCO/Ac2O) after hydrolysis of the glucose unit. X-ray crystallography of the monoacetate showed unambiguously that it had been transformed to a new seco-iridoid having the novel trans diaxial configuration for the protons on C-1, C-9 and C-5. The mechanism for the rearrangement is discussed.

Alkaloids from Crinum moorei.

Thirteen alkaloids were isolated from Crinum moorei two of which are new. These are 3-[4'-(8'-aminoethyl)phenoxy] bulbispermine and mooreine. The structures of the new alkaloids were determined by spectroscopic methods.

Stereostructure and anti-inflammatory activity of three diastereomers of ocobullenone from Ocotea bullata.

A novel diastereomer of ocobullenone. designated as sibyllenone, was isolated from the stem bark of mature Ocotea bullata in the course of a search for anti-inflammatory compounds from this plant. The stereostructure was established by X-ray crystallography and corroborated by NOESY analysis. Ocobullenone, obtained previously, was re-isolated and crystallised successfully for X-ray analysis, thus making possible an accurate spatial comparison of ocobullenone, iso-ocobullenone and the new stereoisomer. Tested pharmacologically for cyclooxygenase-1 and 2, and 5-lipoxygenase inhibition, sibyllenone was the only compound from O. bullata which showed good inhibitory activity towards 5-lipoxygenase.

Analytical and pharmacological investigation of Ocotea bullata (black stinkwood) bark and leaves.

Ocotea bullata (Lauraceae), one of the top-ten traditional medicinal plants used in KwaZulu-Natal, is close to extinction through high demand and destructive harvesting methods. The stem bark is traditionally used to cure headaches, urinary disorders and stomach problems. Substitution of leaves for bark provides a possible resource management solution for this threatened medicinal plant. One aim of this study was to compare the chemical composition of O. bullata leaves and bark using TLC, HPLC and GC-MS analysis. The characteristic analytical fingerprints of leaf and bark extracts showed great similarities. A second aim was to investigate the pharmacological properties of O. bullata as a remedy against headaches. Leaf and bark extracts were tested in terms of cyclooxygenase-1 and 5-lipoxygenase inhibition. Extracts from the bark exhibited moderate inhibitory activity in both test systems. Extracts from fresh leaves were superior to bark extracts in terms of their in vitro inhibitory activity against cyclooxygenase-1 and 5-lipoxygenase. Volatiles obtained from n-hexane extracts of leaves and bark showed better inhibitory activity towards cyclooxygenase-1 and especially towards 5-lipoxygenase than the original n-hexane extracts. Volatiles were therefore recognized as one of the main active principles in O. bullata with regards to the anti-inflammatory properties of this medicinal plant. This lends support to the traditional usage of O. bullata bark as an inhalant or snuff.

An acylated phloroglucinol with antimicrobial properties from Helichrysum caespititium.

A new acylated form of a phloroglucinol with significant antimicrobial properties was isolated by bioactivity guided fractionation from Helichrysum caespititium (Asteraceae). The structure elucidation, and conformation of the new phloroglucinol, 2-methyl-4-[2',4',6'-trihydroxy-3'-(2-methylpropanoyl) phenyl]but-2-enyl acetate, was established by high field NMR spectroscopic and MS data. The compound inhibited growth of Bacillus cereus, B. pumilus, B. subtilis and Micrococcus kristinae at the very low concentration of 0.5 microg/ml and Staphylococcus aureus at 5.0 microg/ml. Six fungi tested were similarly inhibited at low MICs, Aspergillus flavus and A. niger (1.0 microg/ml), Cladosporium chladosporioides (5 microg/ml), C. cucumerinum and C. sphaerospermum (0.5 microg/ml) and Phylophthora capsici at 1.0 microg/ml.

Enolic iridolactone and other iridoids from Alberta magna.

From fresh leaves of Alberta magna two new iridoids and a known cyclopentene dialdehyde have been identified. One of the new compounds has the less-common irido-lactone structure and has an enolic hydrogen on C-4. Two of the compounds show short term mosquito-repellent effects.

A semi-quantitative approach to the selection of appropriate candidate plant molluscicides--a South African application.

The high cost of synthetic molluscicides, used in the control of the intermediate snail hosts of schistosomiasis (bilharzia), has resulted in renewed interest in plant molluscicides. The history of the use of plant molluscicides is reviewed. Although screening programmes have been conducted in other African countries, no efforts have been made to identify South African plants which would be suitable for use locally, using appropriate technology. The prohibitive costs (time and financial) of random surveys for activity necessitated the development of an objective selection procedure. A simple scoring system derived for this purpose is described. Of 600 plant species with potential, or recorded activity, 150 occurred in South Africa. Twenty-six taxa were active according to standards set by WHO. A further 37 species, although untested, warrant further investigation. Species were ranked on cumulative scores for: (a) coincidence of the endemic areas of the plant, snail host and disease; (b) ethnomedicinal value which would provide greater incentive for cultivation; (c) molluscicidal activity (if known, a minimum LD90 of < or = 100 ppm). Two lists resulted, those with recorded and those with potential activity. Both are important in prioritizing research on molluscicidal plants in South Africa. Problems inherent to the scoring system and to the development of plant molluscicides are discussed.

Methylated A-type proanthocyanidins and related metabolites from Cassipourea gummiflua.

The range of natural dimeric A-type proanthocyanidins is extended by identification of two new dimeric afzelechins in the form of 7-OMe-epiafzelechin-(4 beta-->8,2 beta-->0-->7)- epiafzelechin and 7-OMe-epiafzelechin-(4 beta-->8, 2 beta-->0-->7)-ent-afzelechin. These are accompanied in the stem bark of Cassipourea gummiflua by the related metabolites afzelechin, kaempferol-3-0-alpha-L-rhamnopyranoside, afzelechin-3-0- alpha-L-rhamnopyranoside, epiafzelechin (4 beta-->8,2 beta-->0-->7)-epiafzelechin and epiafzelechin-(4 beta-->8, 2 beta-->0-->7)-ent-afzelechin, the latter three compounds being reported from a natural source for the second time.

Structural changes of zeatin during Acid hydrolysis.

Acid hydrolysis of zeatin under reflux at 100°C for 3.5 hours resulted in the formation of two compounds which were active in the soybean callus bioassay. One of these compounds was identified as 6-(3,4-dihydroxy-3-methyl-butylamino)purine. Available evidence suggests that this compound is not likely to be formed when endogenous cytokinins are fractionated with a cation exchange resin such as Dowex 50 at ambient temperatures.

Cytokinins in the Leaves of Ginkgo biloba: I. The Complex in Mature Leaves.

Cytokinin conjugates of zeatin, ribosylzeatin, and their respective dihydro derivatives tentatively have been identified as the major cytokinins present in mature Ginkgo biloba L. leaves. Ribosylzeatin was present in higher levels than zeatin and dihydrozeatin. No evidence could be found that 6-(2,3,4-trihydroxy-3-methylbutylamino)purine occurs as a metabolite in the mature leaves. From the available evidence, it is concluded that cytokinin conjugates are probably the major metabolites formed in the leaves of this deciduous gymnosperm.

(N-methylpyridinium)insulins. Modification at the A19- and B16-tyrosines.

Reaction of 2-iodo-N-methylpyridinium iodide with insulin gave, amongst others, A19- and B16-(N-methylpyridinium)insulin derivatives in which the phenolic groups of the respective tyrosines had been modified. Under carefully controlled conditions the A19 derivative gave small rhombohedral crystals, whereas the B16 derivative failed to crystallize. Both these derivatives had low biological activities.

NA1, NA1, NA1-trimethylinsulin--an insulin analogue with a quaternary amino group at the A1 terminus.

By utilizing the differing reactivity of the amino groups in aqueous organic solvents, des-GlyA1-NB1,N epsilon B29-(Msc)2-insulin was prepared. Its reaction with the phenyl ester of N,N,N-trimethylglycine in the presence of N-hydroxysuccinimide afforded the crystalline NA1,NA1,NA1-trimethylinsulin analogue. In the fat cell assay this analogue has an activity of 49% and, in the mouse convulsion assay, it is 70%.

A crystalline A14-(2-nitro-4-trimethylammoniophenyl) derivative of bovine insulin.

[O-(2-Nitro-4-trimethylammoniophenyl)-TyrA 14]insulin (bovine) is a product formed on reaction of bovine insulin with the hydrophilic reagent 1-fluoro-2-nitro-4-trimethyl-ammoniobenzene iodide (TAN-F) in an aqueous buffer at pH 8.00. The derivative was isolated and its purity established by standard procedures. The identity of the derivative was determined by degrative studies with alpha-chymotrypsin. The addition of zinc to the above reaction decreases the yield of the title derivative, but increases the yield of the [N alpha-TAN-GlyA1] derivative. [N alpha-Boc-GlyA1]insulin was also reacted with the above mentioned reagent in an attempt to improve the yield of the A14-tyrosine derivative. The biological activity of this microcrystalline derivative was found to be 12.4 units/mg as measured by the mouse convulsion assay.