In search of lost ergots: phylogenetic re-evaluation of Claviceps species in Japan and their biogeographic patterns revealed.

Claviceps (Clavicipitaceae, Hypocreales) was erected in 1853, although ergotism had been well-known for a much longer time. By 2000, about 70 taxa had been described in Claviceps, of which eight species and six varieties were based on Japanese type or authentic specimens. Most of these Japanese Claviceps taxa are based on lost specimens or have invalid names, which means many species practically exist only in the scientific literature. The ambiguous identities of these species have hindered taxonomic resolution of the genus Claviceps. Consequently, we sought and collected more than 300 fresh specimens in search of the lost Japanese ergots. Multilocus phylogenetic analyses based on DNA sequences from LSU, TEF-1α, TUB2, Mcm7, and RPB2 revealed the phylogenetic relationships between the Japanese specimens and known Claviceps spp., as well as the presence of biogeographic patterns. Based on the phylogenetic analysis, host range and morphology, we re-evaluated Japanese Claviceps and recognised at least 21 species in Japan. Here we characterised 14 previously described taxa and designated neo-, lecto- and epi-types for C. bothriochloae, C. imperatae, C. litoralis, C. microspora, C. panicoidearum and C. yanagawaensis. Two varieties were elevated to species rank with designated neotypes, i.e. C. agropyri and C. kawatanii. Six new species, C. miscanthicola, C. oplismeni, C. palustris, C. phragmitis, C. sasae and C. tandae were proposed and described. Taxonomic novelties: New species: Claviceps miscanthicola E. Tanaka, Claviceps oplismeni E. Tanaka, Claviceps palustris E. Tanaka, Claviceps phragmitis E. Tanaka, Claviceps sasae E. Tanaka, Claviceps tandae E. Tanaka; New status and combination: Claviceps agropyri (Tanda) E. Tanaka, Claviceps kawatanii (Tanda) E. Tanaka; Typifications (basionyms): Lecto- and epitypification: Claviceps yanagawaensis Togashi; Neotypifications: Claviceps purpurea var. agropyri Tanda, Claviceps bothriochloae Tanda & Y. Muray, Claviceps imperatae Tanda & Kawat., Claviceps microspora var. kawatanii Tanda, Claviceps litoralis Kawat., Claviceps microspora Tanda, Claviceps panicoidearum Tanda & Y. Harada; Resurrection: Claviceps queenslandica Langdon. Citation: Tanaka E, Tanada K, Hosoe T, Shrestha B, Kolarík M, Liu M (2023). In search of lost ergots: phylogenetic re-evaluation of Claviceps species in Japan and their biogeographic patterns revealed. Studies in Mycology 106: 1-39. doi: 10.3114/sim.2022.106.01.

[Superimposition of MR angiography and three dimensional radionuclide brain perfusion image with personal computer].

To investigate the correlation of arterial obstruction with brain perfusion, MR angiography (MRA) and three dimensional (3D) radionuclide brain perfusion image were superimposed. Eleven cases with intracranial artery obstructive patients were studied. Three dimensional brain perfusion images were generated based on the ray-tracing method. Superimposition of MRA on to 3D brain perfusion image was performed on a personal computer. Reconstructing time for 3D image was about 15 minutes for each patient, Superimposing time was about 5 minutes for each image. Correlation of arterial obstruction with decrease in brain perfusion was demonstrated clearly by superimposed image. With a personal computer, it was possible to produce clinically useful synthesized images with relatively short time and conveniently.

Histamine H2-receptor antagonism of T-593: studies on positive chronotropic responses in guinea pig atria.

Histamine H2-antagonistic properties of the novel H2-antagonist T-593, (+-)-N-[2-hydroxy-2-(4-hydroxyphenyl)ethyl]-N'-[2- [[[5-(methylamino)methyl-2-furyl]methyl]thio]ethyl]-N"- (methylsulfonyl) guanidine were investigated on the histamine-induced positive chronotropic responses in isolated guinea pig right atria. T-593 at 3 x 10(-7)-3 x 10(-6) M suppressed the maximal responses of the histamine concentration-response curves in a concentration-dependent fashion, indicating that T-593 is an unsurmountable antagonist. The pD'2 values were 5.50 for T-593 and 5.61 for famotidine; and the IC50 values at 1 x 10(-5) M histamine were 1.05 x 10(-6) M for T-593, 1.59 x 10(-6) M for ranitidine and 1.67 x 10(-7) M for famotidine. T-593 is a racemic compound composed of two enantiomers, (-)-T-593 and (+)-T-593. The histamine H2-antagonistic activity of (-)-T-593 was 1.5-fold more potent than that of racemic T-593, but (+)-T-593 scarcely inhibited the histamine-induced positive chronotropic response. Histamine H2-antagonism by racemic T-593 was mainly attributed to (-)-T-593. Isoproterenol-induced positive chronotropic responses were not affected by T-593 even at 3 x 10(-5) M. Pretreatment of ranitidine for 10 min prior to application of T-593 protected H2-receptors from unsurmountable antagonism by T-593. Reversibility of H2-antagonism was determined every 1 hr after a 30-min treatment of H2-antagonists. T-593 inhibited the positive chronotropic responses for over 6 hr in contrast to fast recovery from inhibition by ranitidine or famotidine. This result showed that T-593 is a slowly dissociable, long-acting histamine H2-antagonist.

[General pharmacology of T-3262, a new pyridonecarboxylic acid].

General pharmacological activities of (+/-)-7-(3-amino-1-pyrrolidinyl)-6-fluoro-1-(2,4-difluorophenyl)-1,4- dih ydro- 4-oxo-1,8-naphthyridine-3-carboxylic acid p-toluenesulfonate hydrate (T-3262), which is a new pyridonecarboxylic acid, were examined with the following results. 1. Central nervous system: T-3262 did not show any significant pharmacological effects at oral doses of 100-1,000 mg/kg but caused slow waves in spontaneous EEG in cats at intravenous doses of 10-30 mg/kg. 2. Respiratory and cardiovascular system: T-3262 produced little effect in normotensive rats and anesthetized rabbits at oral doses of 100-1,000 mg/kg and intravenous doses of 3-30 mg/kg, respectively. But T-3262 caused, dose-dependently, an increase of respiratory rate, hypotension, decrease of heart rate and changes in ECG patterns (elevation of T waves, slow amplitudes of QRS complexes and prolongation of RR interval, etc.) in anesthetized dogs at intravenous doses of 3-10 mg/kg. 3. Renal functions: T-3262 increased electrolyte excretions at oral doses of 300-1,000 mg/kg but did not affect PSP excretion in rats. 4. Autonomic nervous system and smooth muscle organs: T-3262 exerted slight inhibition of gastric output in rats and slight miosis in mice at an oral dose of 1,000 mg/kg. But T-3262 did not affect the contraction of nictitating membrane in anesthetized cats at intravenous doses of 1-30 mg/kg. T-3262 increased spontaneous motilities of isolated stomach, ileum and uterus, but decreased that of isolated colon at concentrations of 10(-5)-10(-4) g/ml. 5. Hematological examinations: T-3262 did not show any significant effects on bleeding time, blood coagulation, platelet aggregation and blood glucose level in rats at oral doses of 100-1,000 mg/kg. 6. Miscellaneous effects: T-3262 exerted slight inhibitions of gastric secretion and of carrageenin-induced hind paw edema in rats at a dose of 1,000 mg/kg administered intraduodenally and orally, respectively. T-3262 did not affect neuromuscular junction and bile secretion in rats at intravenous doses of 3-30 mg/kg and oral doses of 100-1,000 mg/kg, respectively. From these results, it can be assumed that T-3262 has a wide safety margin as an oral antibacterial agent.

[General pharmacology of T-2588, a new oral cephem antibiotic].

A new oral cephem antibiotic, T-2588, the pivaloyloxymethyl ester of (+)-(6R, 7R)-7-[(Z)-2-(2-amino-4-thiazolyl)-2-methoxyiminoacetamido]-3- [(5-methyl-2H-tetrazol-2-yl)methyl]-8-oxo-5-thia-1-azabicyclo[4.2. 0]oct-2-ene-2-carboxylic acid (T-2525), is mainly absorbed from the intestinal tract and biotransformed to T-2525 thereafter. General pharmacological activities of T-2588 were studied and following results were obtained. On the central nervous system, T-2588 did not show any effects at oral doses of 500-2,000 mg/kg and T-2525 produced only a slight elevation of body temperature in rabbits without any other effects at an intravenous dose of 500 mg/kg. On the respiratory and cardiovascular systems, T-2525 caused a slight hypotension and increased both respiratory rate and femoral blood flow, but no changes were observed in heart rate and electrocardiogram in dogs at an intravenous dose of 500 mg/kg. The T-2525 exerted no significant influence on blood pressure response to isoproterenol, acetylcholine or histamine, but showed a slight tendency to decrease pressor response to adrenaline in dogs at intravenous doses of 100-500 mg/kg. For the renal function in rats, T-2588 had no effects on urine volume, electrolytes and PSP excretion at oral doses of 500-2,000 mg/kg. Intravenous administration of T-2525 caused an increase of sodium excretion at 500 mg/kg and dose-dependent increases of PSP excretion at 20-500 mg/kg. Hematological studies revealed that both T-2588 at oral doses of 500-2,000 mg/kg and T-2525 at intravenous doses of 100-500 mg/kg had no effects on bleeding time in mice, blood coagulation and platelet aggregation in rats. The T-2588 exerted no effect on the gastrointestinal system in rats or mice and had no antiinflammatory activity in rats at oral doses of 500-2,000 mg/kg. The T-2525 scarcely affected the motilities of isolated smooth muscle preparations in experimental animals including stomach, ileum, colon, uterus, vas deferens and trachea at a concentration as high as 10(-3) g/ml. The T-2525 increased bile secretion in rats at intravenous doses of 100-500 mg/kg. The T-2525 slightly decreased the twitch tension of musculus gastrocnemius induced by electrical stimulation in rats at an intravenous dose of 500 mg/kg. These results indicate that T-2588 is a pharmacologically inactive antibiotic.

[General pharmacology of T-1982, a new cephamycin antibiotic].

General pharmacological studies on T-1982 produced the following results. On central nervous system, subcutaneous injection of T-1982 at dose of 2,000 mg/kg hastened the onset of pentetrazole-induced tonic extensor in mice. T-1982 had no effect on spontaneous motor activity, pentobarbital hypnosis, body temperature or EEG in mice or rabbits, and also did not show motor incoordinate, anticonvulsive or analgesic activity in mice at intravenous doses of 250--1,000 mg/kg or subcutaneous doses of 500--2,000 mg/kg. On motor and sensory nervous systems, no effect of T-1982 was noted on spinal reflex, neuromuscular junction, conduction anesthesia or surface anesthesia in rats or rabbits. On respiratory, cardiovascular and autonomic nervous systems, T-1982 caused transient increase of respiratory rate, slight hypotension and transient increase of femoral blood flow in dogs at intravenous doses of 250--1,000 mg/kg. However, it caused a slight hypertensive tendency in rabbits. Heart rate and ECG in dogs or rabbits, blood pressure response to epinephrine, isoproterenol, acetylcholine or histamine in dogs, nictitating membrane in cats and pupil size in mice were not affected after intravenous injection of T-1982. No effect was found on isolated guinea pig atrium or rabbit descending aorta following T-1982 application. On renal function in rats, T-1982 caused an increase of PSP excretion but had no effect on urine volume or electrolytes excretion at intravenous doses of 250--1,000 mg/kg. T-1982 prolonged bleeding time in mice at intravenous doses of 500--1,000 mg/kg, but did not show hemolytic property and inhibitory activity on blood coagulation or platelet aggregation in vitro experiments. Spontaneous movement and tone of isolated stomach, ileum, colon, uterus, vas deferens or trachea and acetylcholine-, histamine-, nicotine- or barium chloride-induced contraction of ileum were not affected following T-1982 application. Intestinal propulsion of barium meal in mice, gastric secretion and carrageenin-induced edema in rats were not affected after intravenous injection of T-1982. T-1982 increased bile secretion in rats dose-dependently at intravenous doses of 31.3--125 mg/kg. The local irritative activity of T-1982 in rats was slightly milder than cefoxitin and moderately milder than cefmetazole after intradermal injection. In conclusion, these results suggest that T-1982 would not cause any adverse effects at its estimated clinical doses of 10--20 mg/kg (500--1,000 mg/man).

[General pharmacology of cefoperazone, a new cephalosporin antibiotic (author's transl)].

General pharmacological properties of cefoperazone (CPZ) were studied in various experimental animals. CPZ showed the following effects with intravenous injection to experimental animals. On the central nervous system, CPZ caused vomiting in dogs at 500 mg/kg, pyrexia and slow waves in electroencephalogram in rabbits at 1,000 mg/kg. On the motor and sensory nervous systems, CPZ enhanced slightly the twitch tension of musculus gastrocnemius induced by electrical stimulation in rats at 500 mg/kg. On the respiratory, cardiovascular and autonomic nervous systems, CPZ increased transiently the respiratory rate and potentiated the depressor response to Ach at 125 mg/kg, increased femoral blood flow, potentiated the pressor response to Adr in dogs and decreased the contraction of nictitating membrane induced by electrical stimulation in cats at 500 mg/kg, and then raised the systolic blood pressure in rabbits at 1,000 mg/kg. On the blood, CPZ decreased ChE activity in rabbit plasma at 250 mg/kg, prolonged bleeding time in mice at 500 mg/kg and prothrombin time in rabbits at 1,000 mg/kg. On the smooth muscle organs, CPZ enhanced slightly gastric motility in rabbits and ileal motility in guinea pigs at 62.5 and 125 mg/kg respectively, and promoted gastrointestinal propulsion of BaSO4 meal in mice at 1,000 mg/kg. On the urinary organ, CPZ increased urine volume and electrolytes excretion in dogs at 500 mg/kg. Subcutaneous injection of CPZ scarcely showed any significant effect in experimental animals under the dose of 2,000 mg/kg. In the in vitro experiments, CPZ enhanced slightly the motility of isolated rabbit gastrointestinal tract at 10(-3) g/ml. Assuming the single clinical dose of CPZ should be 10 approximately 40 mg/kg, it is concluded that the occurrence of pharmacological effects observed in experimental animals seems to be very rare clinically.