Enhanced biomass production of duckweeds by inoculating a plant growth-promoting bacterium, Acinetobacter calcoaceticus P23, in sterile medium and non-sterile environmental waters.

Duckweed offers the promise of a co-benefit culture combining water purification with biomass production. Acinetobacter calcoaceticus P23 is a plant growth-promoting bacterium isolated from a duckweed, Lemna aequinoctialis. This study quantified its growth-promoting effect on three duckweeds (L. aoukikusa, L. minor, and Spirodela polyrhiza) in sterile Hoagland solution and evaluated its usefulness in duckweed culture under non-sterile conditions. P23 promoted growth of three duckweeds in sterile Hoagland solution at low to high nutrient concentrations (1.25-10 mg NO3-N/L and 0.25-2.0 mg PO4-P/L). It increased the biomass production of L. aequinoctialis 3.8-4.3-fold, of L. minor 2.3-3.3-fold, and of S. polyrhiza 1.4-1.5-fold after 7 days compared with noninoculated controls. P23 also increased the biomass production of L. minor 2.4-fold in pond water and 1.7-fold in secondary effluent of a sewage treatment plant under non-sterile conditions at laboratory-scale experiments. P23 rescued L. minor from growth inhibition caused by microorganisms indigenous to the pond water. The results demonstrate that the use of P23 in duckweed culture can improve the efficiency of duckweed biomass production, and a positive effect of P23 on duckweed-based wastewater treatment can be assumed.

Different X-linked KDM5C mutations in affected male siblings: is maternal reversion error involved?

Genetic reversion is the phenomenon of spontaneous gene correction by which gene function is partially or completely rescued. However, it is unknown whether this mechanism always correctly repairs mutations, or is prone to error. We investigated a family of three boys with intellectual disability, and among them we identified two different mutations in KDM5C, located at Xp11.22, using whole-exome sequencing. Two affected boys have c.633delG and the other has c.631delC. We also confirmed de novo germline (c.631delC) and low-prevalence somatic (c.633delG) mutations in their mother. The two mutations are present on the same maternal haplotype, suggesting that a postzygotic somatic mutation or a reversion error occurred at an early embryonic stage in the mother, leading to switched KDM5C mutations in the affected siblings. This event is extremely unlikely to arise spontaneously (with an estimated probability of 0.39-7.5 × 10(-28) ), thus a possible reversion error is proposed here to explain this event. This study provides evidence for reversion error as a novel mechanism for the generation of somatic mutations in human diseases.

Age related expression of Werner's syndrome protein in selected tissues and coexpression of transcription factors.

AIMS: Werner's syndrome (WS) is an uncommon autosomal recessive disease resulting from mutational inactivation of human WRN helicase, Werner's syndrome protein (WRNp). Patients with WS progressively develop a variety of aging characteristics after puberty. The aim of this study was to determine the distribution of WRNp and the expression of the transcription factors regulating WRN gene expression in a variety of human organs in an attempt to understand the WS phenotype. METHODS: Tissue specimens were obtained from 16 controls aged from 27 gestational weeks to 70 years of age and a 56 year old female patient with WS. The distribution of WRNp and the expression of the transcription factors regulating WRN gene expression-SP1, AP2, and retinoblastoma protein (Rb)- were studied in the various human organs by immunohistochemical and immunoblot analyses. RESULTS: In the healthy controls after puberty, high expression of WRNp was detected in seminiferous epithelial cells and Leydig cells in the testis, glandular acini in the pancreas, and the zona fasciculata and zona reticularis in the adrenal cortex. In addition, the SP1 and AP2 transcription factors, which regulate WRNp gene expression, appeared in an age dependent manner in those regions where WRNp was expressed. In controls after puberty, SP1 was expressed in the testis and adrenal gland, whereas AP2 was expressed in the pancreas. CONCLUSIONS: These findings suggest that the age specific onset of WS may be related to age dependent expression of WRNp in specific organs.

MR imaging of perinatal brain damage: comparison of clinical outcome with initial and follow-up MR findings.

BACKGROUND AND PURPOSE: The purpose of our study was to determine whether MR studies in the neonatal period are predictive of the neuroradiologic sequelae and clinical outcome in premature and term infants with perinatal brain injury. METHODS: Thirty subjects (15 premature and 15 term infants) with abnormalities revealed by initial MR studies were reexamined approximately 1 year after birth with both MR imaging and a neurologic assessment. All initial MR studies were performed between 35 and 45 weeks corrected age in premature infants and within 28 days of life in term infants. The initial MR studies were evaluated for deep gray matter involvement, hemispheric parenchymal change, intracranial hemorrhage, and periventricular signal and/or morphologic changes. These MR findings were compared with the follow-up MR findings and with the neurologic outcome. RESULTS: The development of cerebral palsy in premature infants was related to the following initial MR findings: subependymal hemorrhage associated with parenchymal destruction, periventricular signal alteration with irregularity of the ventricular wall, and widespread cerebral infarction. These MR findings were predictive of the subtypes of cerebral palsy. In term asphyxiated infants, T2 signal alterations of the deep gray matter rather than T1 shortening and diffuse involvement of the hemispheres were predictive of an unfavorable outcome. Both in term and premature infants, focal hemispheric parenchymal lesions alone (including infarction and intracerebral, subdural, intraventricular, and subarachnoid hemorrhage) did not produce poor outcomes. CONCLUSION: MR studies performed at or near term in either premature or term infants with perinatal brain damage are effective in predicting both late neuroradiologic and clinical outcome.

Development of GABAergic neurons and their transporter in human temporal cortex.

The gamma-aminobutyric acid (GABA) system plays an important role in the early development of the hippocampal formation. The immunohistochemical expression of gamma-aminobutyric acid transporter-1, GAT-1, in the human developing temporal cortex was examined, and the distribution of GAT-1 was compared with that of the 67-kDa isoform of glutamic acid decarboxylase as a marker of GABAergic neurons. Four postmortem tissue specimens from young patients with hippocampal sclerosis were also examined. GAT-1 immunoreactivity was present, with a few puncta, in the neuropil of the stratum oriens and in the molecular layer of the dentate gyrus from 21-22 weeks of gestation, and in the stratum lacunosum-moleculare from 26 weeks of gestation. The peak expression of GAT-1 was seen in early infancy and that of glutamic acid decarboxylase in the perinatal period. These findings may reflect the development of GABAergic inhibitory systems, and may be related to the seizure susceptibility in infancy and early childhood. In the temporal lobes with hippocampal sclerosis, GAT-1 immunoreactivity of the neuropil was preserved in the vicinity of the neuronal loss of the hippocampus. This finding may result from the neurotrophic function of GAT-1 and may be related to its ability of neuronal repair and plasticity in childhood.

Postoperative magnetic resonance imaging of lumbar disc herniation: comparison of microendoscopic discectomy and Love's method.

STUDY DESIGN: We performed a study to compare the magnetic resonance imaging findings up to 24 weeks after microendoscopic discectomy or surgery using Love's method in patients with lumbar disc herniation. OBJECTIVES: The objective was to determine whether or not microendoscopic discectomy was minimally invasive with respect to the nerve roots, cauda equina, and paravertebral muscles by comparing the postoperative magnetic resonance imaging findings in patients treated by microendoscopic discectomy and the conventional Love's method. SUMMARY OF BACKGROUND DATA: We introduced microendoscopic discectomy as a minimally invasive surgical procedure for lumbar disc herniation in September 1998 and have obtained good results. Microendoscopic discectomy is superior to the conventional Love's method in that it reduces postoperative pain, shortens the duration of hospitalization, and allows earlier resumption of normal activities. However, the effect of microendoscopic discectomy on the nerves and paravertebral muscles has not been evaluated objectively. METHODS: Enhancement of the nerve roots and paravertebral muscles, as well as the configuration of the cauda equina at the level of herniation, was assessed on axial magnetic resonance images obtained with contrast enhancement using gadolinium-diethylenetriamine penta-acetic acid before surgery and 1, 4, 8, 12, and 24 weeks after surgery in 25 patients who underwent microendoscopic discectomy and 15 patients who were treated using Love's method. RESULTS: Increased enhancement of the nerve roots was seen in 50.0% of the microendoscopic discectomy group and 46.2% of the Love group at 1 week after surgery. Enhancement of the paravertebral muscles at the surgical site tended to persist for longer in the microendoscopic discectomy group than in the Love group. However, muscle enhancement was widespread in some patients from the Love group. Abnormalities of the cauda equina attributed to surgical invasion were seen in 12.5% of the microscopic discectomy group and 15.4% of the Love group at 1 week after surgery. CONCLUSIONS: Microendoscopic discectomy had an effect on the nerve roots and cauda equina that was comparable with that of Love's method. The magnetic resonance images of the route of entry failed to show that microendoscopic discectomy is appreciably less invasive with respect to the paravertebral muscles.

Vasogenic edema induced by compression injury to the spinal nerve root. Distribution of intravenously injected protein tracers and gadolinium-enhanced magnetic resonance imaging.

The function of the blood-nerve barrier appears quite unique in the nerve root. Protein tracers that were injected into the subarachnoid space passed through the nerve root sheath and entered into the capillary lumen in the endoneurial space but tracers that were injected intravenously did not appear in the endoneurial space. Marked extravasation of protein tracers in the nerve root was induced at the compressed part by strong compression (60 gram force, 30 gram force) and capillaries in the nerve root showed opening of the tight junction accompanied by an increase in vesicular transport under the electron microscope. This situation was reflected as high intensity on Gadolinium-enhanced magnetic resonance imaging. In twenty-one of fifty patients with lumbar disc herniation, the affected nerve root was strongly enhanced by Gadolinium-diethylene-triaminepentaacetic acid, indicating that the blood-nerve barrier in the affected nerve root was broken and intraradicular edema was produced in these cases.

Early immunohistochemical detection of axonal damage and glial activation in extremely immature brains with periventricular leukomalacia.

Extremely low birth weight (ELBW) infants, who died at 12 hours to 7 days after birth, with periventricular leukomalacia (PVL), were examined by means of neuropathological and immunohistochemical methods. Fourteen infants without PVL were used as controls. Anti-beta-amyloid precursor protein (APP), glial fibrillary acidic protein (GFAP), and ionized calcium-binding adaptor molecule 1 (Iba1) antibodies were used as markers for axonal damage, reactive astrocytes and activated microglia, respectively. Thirteen of 14 ELBW infants with PVL showed a widespread distribution of leukomalacia and 10 showed postnatal-onset of leukomalacia. In 12 of the 14 infants with PVL, regions of APP-reactive axons were found multifocally in the cerebral white matter, but 8 of them did not show coagulation necrosis on HE staining. GFAP-positive cells and Iba1-positive cells were markedly found in the white matter of all cases with PVL and slightly in all 14 controls. These results indicated that in ELBW infants, the distribution and formation of PVL foci are widespread and characteristic and so may involve motor and intellectual abilities in ELBW infants. Therefore, the perinatal management to maintain an appropriate cerebral circulation and oxygenation may be very important.

Blood supply of nerve roots and dorsal root ganglia.

The number of patients who suffer from radicular symptoms due to degenerative diseases of the lumbar spine is increasing. This article discusses the differences in anatomic structure between spinal nerve roots and peripheral nerves. Blood supply and cerebrospinal fluid flow are explained, and the effect of mechanical compression, direct and indirect, is shown as a possible cause of symptoms.

Characteristic features of the acetabular labrum in healthy children.

Radial magnetic resonance images of the acetabular labrum were obtained on 40 hips of healthy children. There were no right-left or male-female differences. In children aged 11 years or younger, the labrum on the antero-superior weightbearing portion was triangular in shape, and there was an insular-shaped or linear high-intensity area inside; on the mid-superior portion, the labrum appeared as a regular triangular, low-intensity area; and on the postero-superior portion, it was flat. In children aged 12 and 13 years, the shape of the labrum in each portion was similar to that of the younger children, but the high signal intensity area on the antero-superior portion appeared less frequently. The size of the labrum relative to the femoral head was greater in younger children.

[Obesity and liver dysfunction in UOEH employees--multiple regression analysis of the annual physical checkup data of 1991].

For the purpose of evaluating the significance of obesity as a risk factor toward various chronic geriatric diseases, a multiple regression analysis was performed on the annual physical checkup data of UOEH employees in 1991. The following results were obtained. (1) The average obesity index of the UOEH employees showed a progressive and significant increase in the 10 years from 1981 to 1991. (2) A close relation between the obesity index and serum GPT was recognized by elevation of the standard partial regression coefficients of serum GPT to obesity index and that of the obesity index to serum GPT when the data from all 1591 UOEH employees were analysed in one group. This finding was derived from a significant contribution of obesity to the liver dysfunction in the young male obese population under 30 years of age. (3) Systolic blood pressure was related to age rather than the obesity index, indicating that the development of hypertension is more closely related to aging than obesity. (4) No significant relation was found between the serum total cholesterol level and the obesity index in any group analysed. From the above findings, it can be suggested that the obesity in young male employees is more closely related to liver dysfunction than other abnormalities.

[The significance of obesity in UOEH medical students--multiple regression analysis of the annual physical checkup data in 1991].

For the purpose of evaluating the significance of obesity in a young population as a risk factor toward various chronic diseases, a multiple regression analysis was performed on the data from the annual physical checkup of medical students of UOEH in 1991. The following results were obtained. (1) The average obesity index of the students showed a progressive and significant increase in the past 13 years from 1979 to 1991. (2) A close correlation between the obesity index and serum GPT was recognized by elevation of the standard partial regression coefficient of serum GPT to obesity index and that of obesity index to serum GPT when the data from all 617 students was analysed in one group. This finding was intensified in 142 obese students with an obesity index of over 10%, but it was not seen in the remaining non-obese students. (3) The correlation between the obesity index and serum GPT was found solely in the group of students with constant obesity; i.e., whose obesity index was always more than 10% during a maximum of 9 years in the past. (4) Systolic blood pressure was related to the red blood cell count rather than to the obesity index in this young population. (5) No particularly close correlation to serum cholesterol was found with any of 10 representative items examined in this physical checkup, including the obesity index, indicating that the other factors should be related to serum cholesterol levels. From the above findings, it can be concluded that constant obesity in students is related to liver dysfunction, probably due to fatty liver frequently seen in the precise examination of these individuals.

[Comparison of the casual serum triglyceride levels at health check-up with the values during fasting].

Since higher levels of serum triglycerides were frequently found during the UOEH physical check-up for employees and students in 1989, we analysed subjects with higher triglyceride levels and discussed ways of handling them. Blood sampling to determine serum lipids was done between 13:00-15:00. Of the total (males 1257, females 1267), 272 had higher serum triglyceride levels (over 200 mg/dl) (males 246, females 26) (1st TG). One hundred and twenty-one of the group with higher TG levels in the 1st examination were reevaluated by blood sampling the morning after overnight fasting (2nd TG). There was a significant positive correlationship between the TG levels from the 1st examination and those from the 2nd examination (r = 0.62, n = 121, P < 0.0001). The predictive value calculated from the equation between the TG in the first and second measurements was 341.7 mg/dl. There was a 14.9% rate of false negatives in subjects under 341.7 mg/dl on the first TG measurement. However, there were only 6.3% false negatives in the subjects under 250 mg/dl on the first TG measurement. Age, body mass index and serum cholesterol level were significantly higher in the group with the higher TG level on reevaluation (more than 160 mg/dl) than those in the group with the lower TG level on reevaluation (less than 160 mg/dl). We concluded from these results that the casual serum TG levels above 250 mg/dl might be a suitable cut off level in physical check-ups.

[A case of cardiac amyloidosis showing the ischemic change by exercise Tl-201 myocardial scintigraphy].

A case of cardiac amyloidosis, which was recognized as an ischemic change by exercise Tl-201 myocardial scintigraphy, was studied. The case was 41 year-old woman, whose initial symptom was pretibial edema and died of ventricular fibrillation. Ischemic change was ascertained by the treadmill exercise test and exercise Tl-201 myocardial scintigraphy, however, the significant coronary stenotic lesion was not detected by coronary angiography. Our data suggested that ischemic change may be related with the disturbance of microcirculation caused by deposit of amyloid in the vessels and/or around the vessels from the pathological findings.

[A case of so-called carcinosarcoma of the lung].

A 69-year-old male was followed at our institute for the purpose of hemo-dialization. During follow-up, a tumor shadow (phi 6.5 x 7.0 cm) was found in rt. S5 by chest X-ray. This tumor was diagnosed as squamous cell carcinoma by TBLB. Right middle lobectomy (R 2 a) was performed. Histological staging was pT 2 N 2 M 0, stage IIIa. This tumor was cystic, filled with keratin like substance. Histologically, the cyst wall was composed of well differentiated squamous cell carcinoma and sarcomatous stroma which didn't differentiate into bone, cartilage, muscle, and etc. Immunohistologically, sarcomatous stroma was positively stained by vimentin, negatively by keratin and EMA. This patient survived 7 months with recurrence.

[A case of advanced breast cancer with multiple lung metastases responding well to postoperative combined administration of doxifluridine (5'-DFUR) and tamoxifen].

A 72-year-old female with right breast cancer having multiple lung metastases was treated by combined administration of 5'-DFUR at 800 mg/day, and tamoxifen, at 20 mg/day, after standard radical mastectomy. Plain chest X-ray and computed tomography examinations have shown complete disappearance of the multiple lung tumor since the therapy. No serious side effects have been observed during the treatment course, and there has been no definite evidence of tumor recurrence.

[Consent to enrollment in randomized clinical trials].

Multi-facility joint randomized clinical trials for cancer treatment are the most common method of current clinical study. However, the difference in the rate of participation in multi-facility randomized clinical trials may damage the resulting general validity. Therefore, we studied whether the rate of participation in randomized clinical trials is different between university hospitals and other general hospitals using a questionnaire (with anonymity preserved). There were 744 subjects from university hospitals and 339 from general hospitals participating in the study. The results showed that 10.9 percent of those from university hospitals were willing to participate, against 28.1 percent who were not. Of those from general hospitals (public hospitals in this study) 10.6 percent answered that they would participate and 27.4 percent that they would not. Little difference was found in the rate of participation in randomized clinical trials between university hospitals and other general hospitals. The focus is thus on doctors to solve the possible differences between the facilities in further multi-facility joint studies.