培养温度对多拷贝毕赤酵母猪胰岛素前体的分泌表达和生理特性影响

研究了培养温度对多拷贝毕赤酵母分泌表达猪胰岛素前体（PIP）的生理特性的影响。将多拷贝重组菌A2（12拷贝）和A3（18拷贝）分别在25 ℃和 30 ℃培养,25 ℃培养时PIP表达水平分别比30 ℃培养时提高了40%和32%,而A3细胞的存活率提高了20%。对A3细胞内相关蛋白折叠和氧化程度的KAR2,PDI1, GLR和TRR1基因转录水平分析发现,相比30 ℃培养25 ℃培养时减少了16%~35%,这表明多拷贝重组A3菌株在低温培养时比高温培养在蛋白折叠和氧化协廹响应有明显的减缓效应,从而导致外源蛋白表达提高了40%.     

EUS elastography combined with the strain ratio of tissue elasticity for diagnosis of solid pancreatic masses

Background

Recently, the usefulness of endoscopic ultrasound (EUS) elastography has been reported for the diagnosis of pancreatic lesions. In the present study, we retrospectively assessed EUS elastography as a diagnostic tool by evaluating tissue elasticity distribution and elasticity semiquantification, using the strain ratio (SR) of tissue elasticity, in patients with pancreatic masses.

Methods

One hundred and nine patients who underwent EUS elastography between September 2006 and May 2009 were retrospectively evaluated. The final diagnosis was chronic pancreatitis (CP) in 20 patients [6 with non-mass-forming pancreatitis, 7 with mass-forming pancreatitis (MFP), and 7 with autoimmune pancreatitis (AIP)], pancreatic cancer (PC) in 72, pancreatic neuroendocrine tumor (PNET) in 9, and normal pancreas in 8. The tissue elasticity distribution calculation was performed in real time, and the results were represented in color in fundamental B-mode imaging. In addition, we performed quantification using the SR (non-mass area/mass area).

Results

Elastography for all PC patients showed intense blue coloration, indicating malignant lesions. In contrast, MFP presented with a mixed coloration pattern of green, yellow, and low-intensity blue. Normal controls showed an even distribution of green to red. The mean SR was 23.66?±?12.65 for MFP and 39.08?±?20.54 for PC (P?Conclusions Endoscopic ultrasound elastography is a promising diagnostic tool for defining the tissue characteristics of pancreatic masses. In addition, semiquantitative analysis of elasticity using the SR may allow the differentiation of MFP from PC.     

Characteristics of blood flow in intrauterine growth-restricted fetuses with hypercoiled cord.

OBJECTIVE: To clarify the characteristics of fetoplacental blood flow of growth-restricted fetuses with hypercoiled umbilical cord. SUBJECTS: Eight growth-restricted fetuses with hypercoiled cord. METHODS: Flow velocity waveforms of the umbilical cord artery and vein, fetal abdominal aorta and fetal inferior vena cava were analyzed. RESULTS: The resistance index in the umbilical artery in the hypercoiled cases was lower than that in normal fetuses. Early-diastolic reversed flow was observed in the abdominal aorta in some cases. In all cases, umbilical venous pulsation was observed in the entire cord until delivery. In one case, fetal heart failure occurred, resulting in pre-mature delivery. An atrophic type of single umbilical artery was observed in four cases. CONCLUSION: Fetal blood flow disturbance caused by a hypercoiled umbilical cord may be a cause of growth restriction.     

Complete genome sequence of the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 and comparison with Pyrococcus genomes

The genus Thermococcus, comprised of sulfur-reducing hyperthermophilic archaea, belongs to the order Thermococcales in Euryarchaeota along with the closely related genus Pyrococcus. The members of Thermococcus are ubiquitously present in natural high-temperature environments, and are therefore considered to play a major role in the ecology and metabolic activity of microbial consortia within hot-water ecosystems. To obtain insight into this important genus, we have determined and annotated the complete 2,088,737-base genome of Thermococcus kodakaraensis strain KOD1, followed by a comparison with the three complete genomes of Pyrococcus spp. A total of 2306 coding DNA sequences (CDSs) have been identified, among which half (1165 CDSs) are annotatable, whereas the functions of 41% (936 CDSs) cannot be predicted from the primary structures. The genome contains seven genes for probable transposases and four virus-related regions. Several proteins within these genetic elements show high similarities to those in Pyrococcus spp., implying the natural occurrence of horizontal gene transfer of such mobile elements among the order Thermococcales. Comparative genomics clarified that 1204 proteins, including those for information processing and basic metabolisms, are shared among T. kodakaraensis and the three Pyrococcus spp. On the other hand, among the set of 689 proteins unique to T. kodakaraensis, there are several intriguing proteins that might be responsible for the specific trait of the genus Thermococcus, such as proteins involved in additional pyruvate oxidation, nucleotide metabolisms, unique or additional metal ion transporters, improved stress response system, and a distinct restriction system.     

Transcervical amnioinfusion of antibiotics: a basic study for managing premature rupture of membranes

To determine the best method of preventing ascending infection in the management of premature rupture of membranes, antibiotics such as latamoxef sodium, cefoperazone sodium, and cefotaxime sodium were infused directly into the amniotic cavity in 64 patients undergoing induction of labor at term. A single infusion of 100 or 500 mg of each drug resulted in a concentration of 200 to 1000 micrograms/ml immediately after infusion, and the concentration remained above 10 micrograms/ml for about 24 hours without significant increase in fetal or maternal blood levels. Consequently, a daily single dose of 100 mg or more is probably effective prophylaxis in cases of premature rupture of membranes. When intrauterine infection is suspected, the dose can be increased to 500 mg or more, and transplacental administration may be added to achieve a higher concentration in fetal blood. The present study simulates well premature rupture of membranes, and an amnioinfusion of antibiotics will be reliable and effective in managing premature rupture of membranes.     

Recovery of postural stability following conscious sedation with midazolam in the elderly

Purpose. To investigate the differences in recovery of postural stability, after obtaining similar intravenous sedation levels with midazolam, in elderly and younger patients undergoing dental surgery. Methods. We studied 15 elderly patients (>65 years) and 15 younger patients (<55 years) after intravenous sedation. Midazolam was carefully titrated over 4–5 min until slow response to verbal commands, ptosis of the eyelid, or slight slurring of speech was obtained. Parameters were postural balance tests and an addition test, as a psychomotor function test. Results. The dose of midazolam in the elderly group (0.045 ± 0.012 mg·kg⁻¹) was 62% of that in the younger group (0.074 ± 0.026 mg·kg⁻¹). In evaluation of the percentile rank of a balance test with a visual feedback system, which contained a dynamic balance element, recovery at 60 min in the elderly group was significantly slower than that in the younger group. However, the recovery times for the balance test and the addition test, at which the significantly changed values were restored to the baseline values, were 120 min and 90 min, respectively, in both groups. Conclusion. In the recovery from sedation, elderly patients had more difficulty in acquiring postural adjustment during movement than in maintaining a standing posture. If the dose is carefully administered, however, even elderly patients might be able to return home 2 h after midazolam administration, as could the younger patients. Received: November 6, 2001 / Accepted: April 22, 2002     

Is Bacillus Calmette-Guerin revaccination necessary for Japanese children?

BACKGROUND: Bacillus Calmette-Guerin (BCG) revaccination has been implemented in Japan among tuberculin-negative first grade primary and first grade junior high school students for decades. Controversies regarding the effectiveness of BCG revaccination and low incidence of tuberculosis (TB) among Japanese children prompted this study. METHODS: Cost-effectiveness and cost-benefit analyses were conducted for a cohort of schoolchildren who underwent revaccination during 1996. The study population was a hypothetical cohort comprising 1.35 million first grade primary school and 1.51 million first grade junior high school students enrolled in 1996 at locations throughout Japan. Assuming 50% vaccine efficacy for revaccination, a 10-year duration of protection, and 5% annual discount rate, we calculated the total hypothetical number of TB cases averted, the cost and number of immunizations per TB case averted, and the benefit-cost ratio for the program. RESULTS: The revaccination program for 1996 schoolchildren cohort would prevent 296 TB cases over a 10-year period at a cost of US$ 108,378 per case averted. About 4,963 immunizations would be required to prevent one child from developing TB. The benefit-cost ratio remained at 0.13 with baseline assumptions and ranged from 0.05 to 0.29 and from 0.02 to 0.74 for one-way and two-way sensitivity analyses, respectively. CONCLUSION: BCG revaccination among schoolchildren is not supported by available scientific and economic data. Based on the results of this study, current BCG revaccination policies in Japan and other countries should be reexamined.     

Venoarterial bypass in children with congenital heart disease--a retrospective study

To investigate prognostic factors for patients supported by venoarterial (VA) bypass, we analyzed retrospectively 31 patients with congenital heart diseases supported by VA bypass between 1997 and 2000. Median age was 3.6 months and median body weight was 4.0 kg. Causes of VA bypass were difficult weaning from cardiopulmonary bypass in 8 patients, postoperative cardiac dysfunction in 5 and cardiopulmonary resuscitation in 18. Six (19.4%) of 31 patients were weaned successfully from VA bypass, and 2 (6.5%) survived to hospital discharge. In patients who were weaned from VA bypass successfully, VA bypass was instituted more quickly (41.6 +/- 5.0 vs 62.5 +/- 35.9 min), and good tissue perfusion was established in association with lower lactate levels at 12 hr (7.0 +/- 5.9 vs. 16.4 +/- 15.4 mmol.l-1) and larger urine output during first 24 h (81 +/- 68 vs. 22 +/- 43 ml.kg-1.day-1), compared to those who failed to be weaned. Major complications were intracranial hemorrhage in the newborn (50%), and hypoxic brain damage in patients with palliative operation (35%). It is necessary to establish guidelines of entry, weaning, and withholding of this support.     

Augmentation of the activity of an immunotoxin,anti-Tac(Fv)-PE40KDEL,in T cell lines infected with human T cell leukemia virus type-I

Therapy with an immunotoxin, anti-Tac(Fv)-PE38, which is a conjugate of the variable domains of an anti-Tac monoclonal antibody and Pseudomonas exotoxin, was reported to be useful for adult T cell leukemia (ATL) patients but a considerable amount of the immunotoxin is needed for the therapy and some side effects were also observed. We have previously demonstrated that an immunotoxin, anti-Tac(Fv)-PE40KDEL, showed strong cytotoxic effects on malignant cells from ATL patients. Therefore, we searched for agents that enhance the effects of the immunotoxin. PAK-200, FK-506, quinidine, cepharanthine and cyclosporine A (CsA) augmented the ability of the immunotoxin to inhibit protein synthesis in two human T cell leukemia virus type-I infected T cell lines, KUT-1 and KUT-2. CsA was the most potent agent in both the cell lines. Augmentation of the cytotoxic effect of the immunotoxin by these agents, especially CsA, may be useful in the immunotoxin therapy of ATL.