Use of FM1-43, a membrane-specific fluorescent dye, to estimate plasma membrane integrity in the cryopreservation of green algae.

BACKGROUND AND OBJECTIVE: Cryopreservation of microorganism cultures is an important technology for their use as biological and genetic resources; however, the procedure is complicated and depends on the species. MATERIALS AND METHODS: We used the two-step freezing method for the cryopreservation of the green alga Parachlorella kessleri. RESULTS AND CONCLUSION: The optimal cryoprotectant for cryopreservation was 5% dimethyl sulfoxide plus 5% ethylene glycol. This is different from the optimal cryoprotectant for the closely related species Chlorella vulgaris. Efficient cryopreservation of P. kessleri was achieved using methanol, similar to Chlamydomonas reinhardtii. A membrane-specific fluorescent dye, FM1-43, was applied to estimate plasma membrane integrity. We found that the plasma membrane integrity of P. kessleri cells after freeze-thawing was associated with survivability, suggesting that this is a useful index for the optimization of the first step of the two-step freezing method of cryopreservation.

Oral squamous cell carcinoma with microscopic extracapsular spread in the cervical lymph nodes.

The purpose of this study was to determine the detailed background of cases of oral squamous cell carcinoma (OSCC) with microscopic extracapsular spread (ECS) in the cervical lymph nodes. The cases of 78 patients with primary OSCC, who attended hospital from October 2007 to July 2011 and underwent resection of the primary tumour with neck dissection, were reviewed. The subjects were classified into three categories: pN0, pN+/ECS-, and pN+/ECS+; the outcomes of pN+/ECS+ patients were compared in detail with those of the other categories. Thirty-one cases (39.7%) were pN0, 25 cases (32.1%) were pN+/ECS-, and 22 cases (28.2%) were pN+/ECS+. The 3-year overall survival rate was 82.1% in pN0, 74.1% in pN+/ECS-, and 39.8% in pN+/ECS+ (pN0 vs. pN+/ECS+, P=0.0004; pN+/ECS- vs. pN+/ECS+, P=0.0086). The 3-year disease-specific survival rate was 96.2% in pN0, 77.2% in pN+/ECS-, and 39.8% in pN+/ECS+ (pN0 vs. pN+/ECS+, P<0.0001; pN+/ECS- vs. pN+/ECS+, P=0.0038). Patients with poorly differentiated carcinoma, those with three or more ECS+ nodes, and those with ECS+ node(s) located at levels III, IV, and V, had the worst prognosis among pN+/ECS+ subjects.

Measurement of bone mineral density of lumbar spine and whole body in low-birth-weight infants: comparison of two methods.

In the present study, we compared lumbar spinal and whole-body bone mineral density (BMD) measurements to determine which is more suitable for evaluating the bone mineral status of low-birth-weight (LBW) infants. Lumbar spinal and whole-body BMD were assessed simultaneously in a prospective series including 152 Japanese LBW infants (birth weight 453-2400 g, gestational age 24-38 weeks) from the age of 40 weeks post-conception to 2 years of age. Lumbar spinal BMD at 40 weeks post-conception was significantly correlated with birth weight (r = 0.74; P < 0.0001), but wholebody BMD was not correlated with birth weight. No correlation was found between lumbar spinal and whole-body BMD at 40 weeks post-conception. However, after 40 weeks post-conception, a significant correlation was found between lumbar spinal and whole-body BMD (r = 0.65; P < 0.0001). For infants with a body weight of 4 kg or less at the time of measurement, no correlation was found between lumbar spinal and whole-body BMD. However, for infants with a body weight above 4 kg, a significant correlation was found between lumbar spinal and whole-body BMD (r = 0.65; P < 0.0001). Thus, lumbar spinal BMD is more suitable than whole-body BMD for evaluation of the bone mineral status of LBW in early infancy. Therefore, lumbar spinal BMD should be used for serial evaluation of changes in the bone mineral status of LBW infants.

Bone mineral density of the lumbar spine in very-low-birth-weight infants: a longitudinal study.

UNLABELLED: To examine osteopenia in very low birth weight (VLBW) infants we used repeated dual-energy X-ray absorptiometry in a prospective study of lumbar spinal bone mineral density (BMD) in Japanese VLBW infants (birthweight 426-1498 g; n = 61, group 1) aged 40 weeks postconception to 3 years of age. Control subjects were Japanese infants with birthweight 1500-1999 g (group 2), 2000-2499 g (group 3), or more than 2500 g (group 4). BMD in group 1 during the early period after birth was very low, increased rapidly for 1 year, and then gradually increased until 3 years of age (r = 0.931, P < 0. 0001). BMD at the age of 40 weeks postconception was 0.085 +/- 0.026, 0.132 +/- 0.039, 0.178 +/- 0.042, and 0.196 +/- 0.046 g/cm(2) in groups 1, 2, 3, and 4, respectively (P < 0.0001). However, at 1 and 2 years of age no differences were observed among the groups in BMD. CONCLUSION: This study shows that lumbar spinal BMD in VLBW infants can normalize by the age of 2 years.

Effect of sex steroids on the circulating levels of alpha 2-macroglobulin in injured rats.

The effect of sex steroids on the plasma levels of alpha 2-macroglobulin (alpha 2-M) was investigated to explain the sexual dimorphism observed in response to the injury caused by ip administration of 5 micrograms endotoxin. Mean serum alpha 2-M concentrations were lower in female (46.82 +/- 8.10 arbitrary units) than in male injured rats (82.94 +/- 9.22 arbitrary units). A reduction of plasma alpha 2-M levels was observed after orchidectomy, and the administration of 1 mg testosterone to the same animals increased the response. The same response pattern was observed in ovariectomized rats under the same treatment. The response of androgenized rats (78.93 +/- 12.81 arbitrary units) to injury was similar to the male response. These results suggest the importance of testosterone as the major determinant of this sex-dependent response.

Effect of sex steroids on the circulating levels of alfa2-macroglobulin in injured rats

The effect of sex steroids on the plasma levels of alfa2-macroglobulin (alfa-2M) was investigated to explain the sexual dimorphism observed in response to the injuries caused by ip administration of 5 microng endotoxin. Mean serum alfa2-M concentrations were lower in female (46.82 ñ8.10 arbitrary units) than in male injured rats (82.94 ñ 9.22 arbitrary units). a reduction of plasma alfa2-M levels was observed after orchidectomy, and the administration of 1 mg testosterone to the same animals increased the response. The same response pattern wa observed in ovariectomized rats under the same treatment. The response of androgenized rats (78.93 ñ 12.81 arbitrary units) to injury was similar to the male response. These results suggest the importance of testosterone as the major determinant of this sex-dependent response

[Immunogenicity study of netilmicin (author's transl)].

Immunogenicity of netilmicin (NTL) was studied and following results were obtained. The antisera obtained from rabbits immunized with both NTL alone and NTL . HSA mixture did not show positive response in the heterologous 3-hour PCA reaction and the passive hemagglutination test against the challenge of either NTL alone of NTL . OVA mixture. Guinea pigs immunized with NTL alone did not exhibit systemic anaphylaxis when elicited with NTL alone. The antisera obtained from BALB/c mice immunized with NTL-OVA conjugate adsorbed to A1(OH)3 gel gave positive response in the 24-hour PCA reaction by the method described by MOTA, whereas did not respond to intact NTL in the same system. Similar responses were also obtained in the animals immunized with either gentamicin or gentamicin.protein mixture used as control.

Global and effective synergism of amikacin, gentamicin or tobramycin when combined with carbenicillin against Pseudomonas aeruginosa.

The authors describe a study to evaluate the synergistic effects of the combinations of amikacin-carbenicillin, gentamicin-carbenicillin and tobramycin-carbenicillin at the associated minimal bactericidal concentrations and at doses related to the serum levels reached by the drugs in the blood.