ATP production by sorbitol-treated cells of a methanol yeast, Candida boidinii (Kloeckera sp.) No. 2201

The phosphorylation system of AMP by sorbitol-treated cells of a methanolutilizing yeast, Candida boidinii (Kloeckera sp.) No. 2201, was investigated for the production of ATP. Firstly, reaction conditions for the ATP production were optimized. Under the optimal conditions, 20–30 g 1^?1 of ATP were produced in the conversion rate of 60–70% from AMP. Activities of reactions involved in the ATP-producing system were compared with cells from different cultures to prepare the cells having the higher activity and to know the essential reaction limiting the rate of the system. The energy efficiency of this system was also discussed.     

Specific requirement for inorganic phosphate for induction of bilayer membrane conductance by the cationic uncoupler carbocyanine dye

The trinucleous divalent cationic cyanide dye triS-C₄(5) was shown to be an uncoupler of oxidative phosphorylation in mitochondria only in reaction medium containing inorganic phosphate (P_i). This dye also induced marked increase in the electrical conductance of a phospholipid bilayer membrane in bathing solution containing P_i, but not in solution containing Tris-HCI buffer without P_i. Time-dependent fluctuation of the electrical current across the bilayer membrane was observed in the presence of triS-C₄(5) only in bathing solution containing P_i. This fluctuation could be due to perturbation of the bilayer membrane structure induced by the cooperative action of the cyanine dye and P_i, and this perturbation should be directly related to their effects in increasing membrane conductance and also causing uncoupling in mitochondria.     

Use of a dual priming oligonucleotide system to detect multiple sexually transmitted pathogens in clinical specimens

Aims:  To evaluate a new dual priming oligonucleotide (DPO)-based multiplex polymerase chain reaction (PCR) assay for detection of six sexually transmitted pathogens, including Chlamydia trachomatis , Neisseria gonorrhoeae , Mycoplasma genitalium , Mycoplasma hominis , Ureaplasma urealyticum and Trichomonas vaginalis .
Methods and Results:  Using 130 clinical specimens, the results obtained by the multiplex PCR, previously established in-house PCR and COBAS Amplicor PCR assays were compared. The specimens frequently contained multiple pathogens (34/130 specimens). The multiplex PCR assay had an overall sensitivity of 96% and specificity of 100% compared to the in-house PCR assay at >20 μg ml⁻¹ of DNA concentrations in samples and there was no cross-reaction with nonpathogenic Neisseria species that cause the majority of false-positive results with the COBAS Amplicor PCR assay.
Conclusions:  The DPO-based multiplex PCR assay detected the six sexually transmitted pathogens in clinical specimens with a high sensitivity and specificity, although its sensitivity was dependent on the DNA content of the samples.
Significance and Impact of the Study:  It is the first report about the new DPO-based technique to detect multiple sexually transmitted pathogens in a single assay, which has considerable potential to diagnose the infections accurately and rapidly.     

Rhabdosargus niger (Perciformes: Sparidae), a new sparid species from Indonesia, with taxonomic status of the nominal species synonymized under Rhabdosargus sarba

A new sparid Rhabdosargus niger is described on the basis of three type specimens collected from off Kalimantan, Indonesia. The species can be differentiated from all known congeners by the following combination of characters: dorsal-fin rays XI, 13, anal-fin rays III, 11, scale rows below lateral line 11½–12½, scale rows between the fifth dorsal-fin spine base and lateral line 5½, pored lateral-line scales 53–55, head and body silvery-black, pelvic and anal fins dusky gray, and no golden longitudinal lines on the body. Partial sequences of mitochondrial COI gene (571 bp) strongly supported the distinctiveness of R. niger within the genus. Taxonomic status of the nominal species synonymized under Rhabdosargus sarba (Forsskål 1775) is discussed and the key to the species of Rhabdosargus is provided.     

Transgenic expression of group V, but not group X, secreted phospholipase A2 in mice leads to neonatal lethality because of lung dysfunction

In an effort to elucidate the functions of secreted phospholipase A2 (sPLA2) enzymes in vivo, we generated transgenic (Tg) mice for group V sPLA2 (sPLA2-V) and group X sPLA2 (sPLA2-X), which act potently on phosphatidylcholine in vitro.We found that sPLA2-V Tg mice died in the neonatal period because of respiratory failure. The lungs of sPLA2-V Tg mice exhibited atelectasis with thickened alveolar walls and narrow air spaces, accompanied by infiltration of macrophages and only modest changes in eicosanoid levels. This severe pulmonary defect in sPLA2-V Tg mice was attributable to marked reduction of the lung surfactant phospholipids, phosphatidylcholine and phosphatidylglycerol. Given that the expression of sPLA2-V is greatly elevated in human lungs with severe inflammation, our present results raise the intriguing possibility that this isozyme may contribute to ongoing surfactant hydrolysis often observed in the lungs of patients with respiratory distress syndrome. In contrast, sPLA2-X Tg neonates displayed minimal abnormality of the respiratory tract with normal alveolar architecture and surfactant composition. This unexpected result was likely because sPLA2-X protein existed as an inactive zymogen in most tissues. The active form of sPLA2-X was detected in tissues with inflammatory granulation in sPLA2-X Tg mice. These results suggest that sPLA2-X mostly remains inactive under physiological conditions and that its proteolytic activation occurs during inflammation or other as yet unidentified circumstances in vivo.     

Proteomic analysis of secreted proteins from aseptically grown rice

Plants are known to secrete a variety of compounds into the rhizosphere. These compounds are thought to play important roles in the regulation of soil chemical properties and soil microorganisms. To determine the composition of proteins secreted from rice roots, aseptic hydro culture was performed, and the collected proteins were analyzed. Over 100 proteins were identified; most were identified using the rice database (RAP-DB), and about 60% of the identified proteins were suspected to have a signal peptide. Functional categorization suggested that most were secondary metabolism- and defense-related proteins. Pathogenesis- and stress-related proteins were the major proteins found in the bathing solution under aseptic conditions. Thus, we propose that rice plants constitutively secrete a large variety of proteins to protect their roots against abiotic and/or biotic stresses in the environment.