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Batrachotoxin is an extremely potent cardio- and neurotoxic steroidal alkaloid found in certain species of frogs, birds, and beetles. The steroidal 6/6/6/5-membered carbocycle (ABCD-ring) is U-shaped and functionalized with two double bonds, a six-membered C3-hemiacetal across the AB-ring, a seven-membered oxazepane on the CD-ring, and a dimethylpyrrolecarboxy group at the D-ring carbon chain. These structural features present an unusual and formidable synthetic challenge. Herein we report a total synthesis of batrachotoxin based on a newly devised convergent strategy through a 22-step sequence. Enantiopure AB-ring and D-ring fragments were prepared and subjected to a crucial C(sp 2 )-C(sp 2 ) coupling reaction. Although both C(sp 2 ) centers were sterically encumbered by proximal tetrasubstituted carbon atoms, Ag 2 O strongly promoted the Pd(PPh 3 ) 4 -catalyzed Suzuki-Miyaura coupling reaction at room temperature, thereby connecting the two fragments without damaging their preexisting functionalities. Subsequent treatment with t-BuOK induced Dieckmann condensation to cyclize the C-ring. The judiciously optimized functionalizations realized oxazepane formation, carbon chain extension, and pyrrole carboxylic acid condensation to deliver batrachotoxin., (© 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Kikai submarine caldera to the south of the Kyushu Island, SW Japan, collapsed at 7.3 ka during the latest supereruption (>500 km 3 of magma) in the Japanese Archipelago. Multi functional research surveys of the T/S Fukae Maru in this caldera, including multi-beam echosounder mapping, remotely operated vehicle observation, multi-channel seismic reflection survey, and rock sampling by dredging and diving, provided lines of evidence for creation of a giant rhyolite lava dome (~32 km 3 ) after the caldera collapse. This dome is still active as water column anomalies accompanied by bubbling from its surface are observed. Chemical characteristics of dome-forming rhyolites akin to those of presently active small volcanic cones are different from those of supereruption. The voluminous post-caldera activity is thus not caused simply by squeezing the remnant of syn-caldera magma but may tap a magma system that has evolved both chemically and physically since the 7.3-ka supereruption.

Foscarnet (trisodium phosphonoformate, PFA) is an effective inhibitor of retroviral reverse transcriptase (RT) and is known to block the replication of human immunodeficiency virus type 1 (HIV-1). In this article we analyzed the evolutionary process in generating HIV-1 strains related to drug resistance, using PFA as a selective pressure. PFA inhibited virus replication and protected the virus-induced cell killing, but it did not completely eliminate HIV-1 during the course of 7 weeks of treatment. The nucleotide sequence of the 859-bp DNA fragment spanning the core region of the HIV-1 pol gene was determined for 51 clones obtained from genomic DNA of the HIV-1-infected cells at different time points during PFA treatment. The nucleotide sequence analysis documented the presence of a minor HIV-1 variant prior to the PFA treatment. Molecular evolutionary techniques were utilized to analyze how the minor HIV-1 clones became predominant during this evolutionary process under the selective pressure of PFA. A phylogenetic tree analysis divided these 51 HIV-1 clones into 3 groups. One of the groups consisted of the clones associated with the resistance to PFA. The clones belonging to this group became predominant over time during the course of PFA treatment. Thus, the acquisition of PFA resistance by HIV-1 was considered to be due to clonal selection. Furthermore, among the various amino acid substitutions observed, the substitution of arginine at position 172 by lysine (Arg172Lys) clearly distinguished this group from the others. Since the consistent amino acid substitution observed here has not been identified in the HIV-1 strains resistant to other RT inhibitors, PFA in combination with other RT inhibitors is considered to be a feasible candidate for a convergent combined chemotherapy against HIV-1 in the treatment of patients with AIDS and related conditions.

A total of 132 strains of V. parahaemolyticus isolated from patients and from the suspected causal food items of past food poisoning cases occurring in Kanagawa Prefecture, Japan, were examined for the ability to hydrolyze urea, with specific reference to the presence of the thermostable direct hemolysin gene (tdh) and the gene for thermostable direct hemolysin-related hemolysin (trh). Ten strains belonging to five different O-antigen serotypes were positive for urea hydrolysis (UH+), and four of these strains did not carry tdh. A total of 106 strains carried tdh, but less than 6% of them were UH+, whereas all trh-carrying strains were UH+. The evidence suggests that urea hydrolysis is not a reliable marker for identifying tdh-carrying V. parahaemolyticus strains in Japan (the Pacific Northeast) but may be a marker for trh-carrying strains.

To evaluate DNA fingerprinting as an epidemiologic tool, pulsed-field gel electrophoresis (PFGE) was performed on isolates of Salmonella, including S. typhimurium, S. thompson, and S. enteritidis. Chromosomal DNA was digested with the restriction endonucleases Bln I and Xba I. The patterns of S. thompson and S. typhimurium isolates from various sources were different from one another. There was no correlation between the phage type and the digestion pattern of S. enteritidis isolates. Some strains belonging to one phage type were distinguished by their PFGE pattern in this study. These results suggest that the Bln I and Xba I digestion patterns of chromosomal DNA are useful for epidemiological analysis of an outbreak of Salmonella infection or food poisoning.

During molecular cloning of full-length retroviral plasmid clones occurrence of homologous recombination (HR) between LTR regions is frequently observed. In order to evaluate appropriate host bacterial strains for cloning such HR-prone plasmids, we utilized a linearized template plasmid containing a full-length HIV-1 proviral sequence. The plasmid was linearized within the viral sequence so that plasmid transformed bacteria would grow only when the plasmid was circularized by HR. Using this genetic system for detecting HR, we evaluated the frequency of HR in various recA(-) bacterial strains which are commercially available and in some recA-null strains in which recA-defective phenotype was constructed by P1 transduction. We found that HR occurred even in recA-null strains although in lesser frequencies. The nucleotide sequence analysis at the junction of recombination revealed no loss, insertion or duplication of DNA sequence. It is suggested that recombination machinery other than the RecA system is involved.

Survival of five strains of Vibrio cholerae, including serotypes O139 Synonym Bengal, O1 El Tor, and non-O1 were compared in water from a river. These bacteria were mixed with water from a river and the water filtered through 0.45 micron millipore filters, respectively, to yield a concentration of 10(6) CFU/ml and incubated at 5 degrees C and 20 degrees C for 21 days. The survival curve of V. cholerae O139 was almost the same with V. cholerae O1 and non-O1. The number of these bacilli decreased to less than 10(2) CFU/ml at 7 days of the incubation at 20 degrees C. When incubated at 5 degrees, however, these bacilli survived much longer and the number decreased to the same value after 14 days. Therefore, these results indicated that the temperature during the incubation greatly affects the survival of V. cholerae. On the other hand, when the water was filtered and used for the experiment, V. cholerae survived longer than in the polluted water. From these observations, if the river is polluted with V. cholerae O139 in the future, it is suggested that the distribution of V. cholerae O139 in the river may be the same as the present condition with V. cholerae O1 and non-O1.