Mycoplasma-like Organisms Associated with Proliferation and Big Bud of Solanum lycocarpum St. Hil.

Mycoplasma-like organisms (MLOs) were found in sieve tubes of Solanum lycocarpum affected with prolferation and big bud. The MLOs were graft-transmitted to tomato plants which developed similar symptoms. The association of MLOs with affected but not with symptomless S. lycocarpum and tomato suggests that the abnormality is induced by MLOs. The possible role of S. lycocarpum as a reservoir of a strain of the tomato big bud is discussed.     

Evaluation of Gamma-Radiation Inactivation of a Bioterrorism Agent, <Emphasis Type="Italic">Bacillus anthracis</Emphasis> Spores,on Different Materials

Decontamination of suspected packages, such as sealed envelopes, liquids and tools that are likely contaminated with biological agents is of great importance. In this study, we aimed to determine the gamma radiation dose required for the decontamination of paper, fabric and liquid materials without causing any damage to the structure of these materials. Each study group included 11 pieces of paper, fabric and sterile saline contaminated with 0.8 × 10⁵ virulent Bacillus anthracis (B. anthracis) spores. These specimens were exposed to doses of 5.49, 11.58, 17.21, 21.75, 27 and 33.1 kilogray (kGy) of gamma radiation from a cobalt-60 source. After irradiation of all the samples, a viability assessment of the B. anthracis spores was performed. It was found that full decontamination was achieved with 11.58 kGy on the paper samples and 17.21 kGy on the fabric and liquid samples. It was concluded that a dose of 20 kGy of gamma radiation may be recommended for the inactivation of B. anthracis for some surfaces when especially sensitive and valuable materials cannot be wet decontaminated were exposed. In addition, serologic and molecular assays of the suspected packets can be performed for forensic purposes without damaging existing evidence in a bioterror incident.     

Agrobacterium‐mediated transformation of Vitis Cv. Monastrell suspension‐cultured cells: Determination of critical parameters

Although some works have explored the transformation of differentiated, embryogenic suspension‐cultured cells (SCC) to produce transgenic grapevine plants, to our knowledge this is one of the first reports on the efficient transformation of dedifferentiated Vitis vinifera cv Monastrell SCC. This protocol has been developed using the sonication‐assisted Agrobacterium‐mediated transformation (SAAT) method. A construct harboring the selectable nptII and the eyfp/IV2 marker genes was used in the study and transformation efficiencies reached over 50 independent transformed SCC per gram of infected cells. Best results were obtained when cells were infected at the exponential phase. A high density plating (500 mg/dish) gave significantly better results. As selective agent, kanamycin was inefficient for the selection of Monastrell transformed SCC since wild type cells were almost insensitive to this antibiotic whereas application of paromomycin resulted in very effective selection. Selected eyfp‐expressing microcalli were grown until enough tissue was available to scale up a new transgenic SCC. These transgenic SCC lines were evaluated molecularly and phenotypically demonstrating the presence and integration of both transgenes, the absence of Agrobacterium contamination and the ability of the transformed SCC to grow in highly selective liquid medium. The methodology described here opens the possibility of improving the production of valuable metabolites. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:725–734, 2016     

Zur genetik der 6-Phosphogluconatdehydrogenase (EC:1.1.1.44) bei Säugern

Zusammenfassung Das Enzym 6-PGD zeigt bei den verschiedensten Species einen genetisch gesteuerten Polymorphismus, der sich bei elektrophoretischer Darstellung in einer Variabilität des Isoenzymmusters äußert. Die Befunde bei den bislang untersuchten Säugern unter Einschluß des Menschen lassen sich formalgenetisch nach dem Modell: 1 Genlocus mit multipler Allelie interpretieren. Zur Prüfung der Frage, ob das Säugergenom einheitlich nur ein Gen für dieses Enzym enthält oder ob bei einzelnen Species eine Duplikation dieses Gens existiert, wurdene eine Anzahl Säuger verschiedener Ordnungen untersucht. In einer ersten Mitteilung wird hier über 6 verschiedene Species der Microtinae berichtet. Die Befunde sprechen durchweg für die Existenz von jeweils nur 1 Genlocus der 6-PGD. Bei Microtus oeconomus wurden 2 Allele an diesem locus nachgewiesen, bei Microtus ochrogaster 3 verschiedene Allele. Durch Familienuntersuchungen bei diesen beiden Species werden alle möglichen Kombinationen dieser Allele nachgewiesen.

---

The genetics of 6-PGD (EC: 1.1.1.44) in various mammals I. Studies on 6 species of Microtinae, Rodentia. Isoenzyme polymorphism and family studies in Microtus oeconomus and Microtus ochrogaster

Summary The enzyme 6-PGD exhibits a genetically determined polymorphism which results in a variability of the isoenzyme pattern. In mammals so far examined the findings are to be interpreted according to the model of multiple alleles at one single gene locus. In order to test whether the mammalian genome is uniformly endowed with one single locus for this enzyme or whether a duplication of this locus is present in certain species, we examined various mammals of different orders. Our results in this first communication on 6 different Microtinae species are all in agreement with the assumption of a single 6-PGD locus. In Microtus oeconomus 2 alleles could be demonstrated and in Microtus ochrogaster 3 different alleles. Family studies performed with both species revealed the occurrence of all the expected combinations of these alleles.

     

Identification of novel α7 nicotinic receptor ligands by in silico screening against the crystal structure of a chimeric α7 receptor ligand binding domain

A hierarchical in silico screening procedure using the crystal structure of an agonist bound chimeric α7/Ls-AChBP protein was successfully applied to both proprietary and commercial databases containing drug-like molecules. An overall hit rate of 26% (pK_i ?5.0) was obtained, with an even better hit rate of 35% for the commercial compound collection. Structurally novel and diverse ligands were identified. Binding studies with [³H]epibatidine on chimeric α7/5-HT₃ receptors yielded submicromolar inhibition constants for identified hits. Compared to a previous screening procedure that utilized the wild type Ls-AChBP crystal structure, the current study shows that the recently obtained α7/Ls-AChBP chimeric protein crystal structure is a better template for the identification of novel α7 receptor ligands.     

Inhibition of ATP dephosphorylation by acaricides with emphasis on the anti-ATPase activity of the carbodiimide metabolite of diafenthiuron.

3-(2,6-Diisopropyl-4-phenoxyphenyl)-1-tert-butylcarbodiimide (DFCD), a toxic metabolite and photodegradation product of the propesticide diafenthiuron, and dicyclohexylcarbodiimide (DCCD), a commonly used biochemical inhibitory probe, inhibited Mg(2+)-, Na+K(+)-, and Ca2+Mg(2+)-ATPase activities in preparations from bulb mites (Rhizoglyphus echinopus (Fumouze and Robin)), twospotted spider mites (Tetranychus urticae Koch), and bluegill (Lepomis macrochirus Rafinesque) brain. DCCD was more active than DFCD, but neither carbodiimide was very potent. A possible exception occurred with DCCD, which caused 100% inhibition of bulb mite oligomycin-sensitive Mg(2+)-ATPase activity at a concentration of 0.1 mM. Using house fly, Musca domestica L., thorax mitochondrial Mg(2+)-ATPase, we showed that the binding domain for both carbodiimides was in the F0 portion of the enzyme, probably the transmembrane proton channel which is the known site of DCCD binding in proton-translocating ATPases. Certain other specific acaricides were inhibitors (greater than 50% at 0.1 mM) of ATPase preparations from bulb mites. These acaricides included chloropropylate, bromopropylate, oxythioquinox, cyhexatin, and flubenzimine (Mg2+ and Na+K(+)-ATPase), and ovex, chlorbenside, and propargite (Mg(2+)-ATPase). The role of ATPase inhibition in the modes of acaricidal and insecticidal actions of diafenthiuron, the two carbodiimides, and the other compounds is discussed.     

Antiproliferative and newly attributed apoptotic activities from an invasive marine alga: Caulerpa racemosa var. cylindracea

Caulerpa racemosa (Forsskål) is a green marine alga which spreads from tropical to warm-water regions. Due to having invasive capacity C. racemosa var. cylindracea is a well-known biological pollution in Mediterranean Sea. One of the most important secondary metabolites of C. racemosa is Caulerpenyne (CPN). In the present study, antiproliferative and apoptotic effects of C. racemosa var. cylindracea extract and purified CPN on two well-known neuroblastoma cell lines, SHSY5Y and Kelly, are investigated. The antiproliferative and, additionally, newly attributed apoptotic effects of both C. racemosa var. cylindracea extract and purified CPN on SHSY5Y and Kelly cell lines have been shown in the present study. IC₅₀ values are 0.59 ± 0.06; 1.06 ± 0.23 g wet alga/methanol and 5.64 ± 0.09; 6.02 ± 0.09 μM CPN for C. racemosa var. cylindracea extract and purified CPN on SHSY5Y and Kelly cell lines, respectively. Percentages of apoptotic cells of SHSY5Y and Kelly in 0, 0.1 and 1 μM CPN conditions were 1.00 ± 0.71, 3.00 ± 0.71 and 49.40 ± 3.78, 39.60 ± 6.19 and 78.00 ± 2.74, 69.40 ± 3.78, respectively. In conclusion, the present study shows the antiproliferative effect of C. racemosa var. cylindracea extract and newly attributed apoptotic effects of C. racemosa var. cylindracea this extract. Compared to other alkylating anticancer drugs, CPN and also C. racemosa var. cylindracea extract might be considered as an alternative native source of antitumor drugs. Inasmuch as both C. racemosa extract and CPN have shown both antiproliferative and apoptotic effects on SHSY5Y and Kelly cell lines, the CPN and CPN derivatives might be considered as multifunctional agents in cell metabolism.