Effects of 3′-C-Methylation on the Hydrolytic Stability and Hydroxyl pK a Values of Dinucleoside 2′,5′- and 3′,5′-Monophosphates

Abstract

The first-order rate constants for hydrolysis of 3′-C-methyluridylyl(2′,5′)- and -(3′,5′)adenosine and the corresponding native dinucleoside monophosphates (2′,5′- and 3′,5′-UpA) have been determined as a function of hydroxide-ion concentration (0.025 - 7 M) at 25°C. In addition to the effects on the hydrolytic stability of the compounds, the effects of the 3′-C-methyl substitution on the kinetically determined pK _a values for the sugar hydroxyls of the undine moiety are discussed.     

Cleavage of the Phosphodiester Bond of Uridylyl-(3′,5′)-8-Carboxymethylaminoadenosine by Hydronium,Hydroxide and Zinc(Ii) Ions: A Model Study Aimed at Elucidating the Potential of a Carboxylate Function as an Intramolecular Catalyst

Abstract

Uridylyl-(3′,5′)-8-carboxymethylaminoadenosine has been synthesised, and its transesterification to uridine 2′,3′-cyclic phosphate in the presence and absence of Zn²⁺ ion has been studied. The results show that a carboxylate function in the vicinity of the phosphodiester bond accelerates the metal ion promoted cleavage but not the metal ion independent reaction. Under acidic conditions, the predominant reaction is the cleavage of the side chain, giving the 8-amino derivative.     

Hydrolytic Reactions of 3′-Deoxy-3′-thioinosylyl-(3′→ >5′)-uridine; An RNA Dinucleotide Containing a 3′-S-Phosphorothiolate Linkage

Abstract

The course of hydrolysis of 3′-deoxy-3′-thioinosylyl-(3′ → 5′)-uridine (IspU) has been followed by HPLC over a wide pH-range. Two reactions of the internucleosidic thiophosphate linkage compete: (i) cleavage yielding thioinosine monophosphates and uridine, and (ii) isomerization to the 2′,5′-isomer of IspU. Under very acidic conditions, even acid-catalyzed depurination of the inosine moiety is observed. The stability of the thiophosphate linkage and the mechanisms of its rupture are discussed.     

Persistence of Trichinella spiralisin Rat Carcasses Experimentally Mixed in Different Feed

Trichinella spiralis infected rat carcasses were incubated for 6 weeks in several animal feeds to assess how long Trichinella can present a risk for an outbreak in contaminated feeds. In groups of 6, 24 infected target rats were placed in silage, grained barley, propionic acid-preserved feed, and also into simulated pasture conditions. Test environments were sampled after one-, 2-, 4-, and 6-week-incubations. Trichinella larvae were recovered by digestion, and their infectivity was evaluated in rats. A two-week incubation reduced the number of recovered larvae, but still after 6 weeks low numbers were isolated from all feeds except from the experimental group simulating pasture conditions. After 2 weeks storage, the larvae were infective in all storage environments. However, up to 4 weeks, they survived only in the propionic acid-fermented feed and there in small numbers with reduced reproductive capability. This indicates the possibility of farm animals to get infection from rats or other infected material being hazardously mixed with hay or other feed. If silage is stored for at least one month before use, however, the risk from this forage appears to be minimized.     

A Comparative Study on the Cleavage of Stereoisomeric Uridylyl (3',5')Uridines [D,D-, D,L- and L,D-UPU] by Acid,Base and Metal Ion Catalysts

Stereoisomeric uridylyl(3',5')uridines D,L-UpU andL,D-UpU were synthesised. Their cleavage was followed in thepresence of acid, base and metal ion catalysts to studywhether the stereochemistry affects the inherent reactivity ofthe internucleosidic phosphodiester bond, and whether the lowmolecular weight catalysts can distinguish between thesubstrates. The rate constants obtained were compared to thoseof D,D-UpU. The comparison shows that the stability of thephosphodiester bond does not depend on the stereochemistry ofthe sugar rings. In contrast slight reactivity differences areobserved in the presence of metal ion catalysts, whichsuggests that selective cleavage of stereoisomeric substrateseven by small molecular weight chemical catalysts may bepossible.     

On the Structure and Reactivity of Y-Nucleoside [Wyosine]

Abstract

Wyosine-Triacetate 4 has found to undergo electrophilic reactions exclusively at the C-7 position. ¹⁵N-NMR of 4, shows that the primary site of protonation is N-5 suggesting that the “right” imidazole ring is more aromatic that the “left” imidazole moiety.     

Associations between Trichinella species and host species in Finland

Examination of 627 wild animals--raccoon dogs (Nyctereutes procyonoides), red foxes (Vulpes vulpes), European lynxes (Lynx lynx), brown bears (Ursus arctos), wolves (Canis lupus), and badgers (Meles meles)--revealed Trichinella spp. The prevalence varied according to geographical region of Finland (north; southwest, SW; and southeast, SE) and was the highest among lynxes (70%, SW). The risk of trichinellosis was higher in the SE (odds ratio, OR, 19.4) and SW regions (OR 14.3), as compared with the northern region (OR 1), with no difference between the former 2 regions. Foxes (OR 2.1) and lynxes (OR 1.9) had a higher risk than raccoon dogs (OR 1) of being infected. The distribution of different Trichinella species was evaluated in 87 wild and domestic mammals by multiplex polymerase chain reaction. Trichinella spiralis was detected more often in domestic and synanthropic animals than in sylvatic hosts. Trichinella nativa was detected only in wildlife. Trichinella pseudospiralis was found both in sylvatic and synanthropic hosts. Trichinella britovi was detected only in mixed infections with other Trichinella species. The raccoon dog was the sole host for all 4 Trichinella species and also carried the most intense infections.     

HPLC as a Tool to Study the Kinetics of Parallel and Consecutive Reactions: Competitive Interconversion,Dephosphorylation, Deamination and Depyrimidination of Cytidine 2′- and 3′-Monophosphates in Aqueous Acid

Abstract

The applicability of HPLC as a method to study the kinetics of complicated reaction systems of nucleosides and nucleotides has been demonstrated by using the hydrolytic reactions of cytidine 2′- and 3′-monophosphates as an example.     

Preparation,Hydrolysis and Intramolecular Transesterification of 3′-Deoxy-3′-thioinosine 3′-S-Dimethylphosphorothiolate

Abstract

The hydrolytic reactions of the dimethyl ester of 3′-deoxy-3′-thioinosine 3′-S-phosphorothiolate have been followed over a wide aciditty range by HPLC. At pH > 3, only hydroxide ion catalyzed isomerization to the 2′-dimethylphosphate takes place, whereas under more acidic conditions hydrolysis to the 2′-monomethylphosphate and 3′-S-monomethylphosphorothiolate competes. The latter is the only product accumulating in very acidic solutions (1 M hydrochloric acid). Mechanisms of the reactions are discussed.     

Experimental trichinellosis in reindeer

Six female reindeer calves were inoculated intraruminally with various doses of Trichinella muscle larvae. Four calves were inoculated with T. nativa, receiving 15,000 (n = 1), 5,000 (1), and 2,500 (2) larvae each. Two calves were inoculated with 5,000 T. spiralis larvae each. Blood samples were collected twice per week for total white blood cell (WBC) and differential counts and for serology using enzyme-linked immunosorbent assay (ELISA) based on T. spiralis excretory-secretory antigen. On day 56, the calves were slaughtered and muscle samples were examined according to the standard digestion method for Trichinella larvae. Blood samples were also collected twice a week from 4 uninoculated, but otherwise similar, reindeer calves corralled separately. Both the total WBC and eosinophil counts of the inoculated animals were, on average, higher during the experimental period. All the inoculated calves seroconverted, showing an increase in the optical density (OD) in the ELISA starting between day 23 and day 27 postinoculation. Very few muscle larvae (<0.08 larvae/g [lpg]) were to be found from the animals inoculated with T. nativa, but about 4 and 6 lpg were recovered from the masseter muscles of those inoculated with T. spiralis.