Subchronic (13-week) oral toxicity study, preceded by an in utero exposure phase, with arachidonate-enriched triglyceride oil (SUNTGA40S) in rats.

Polyunsaturated fatty acids (PUFAs), such as arachidonic acid (ARA) and docosahexaenoic acid (DHA) are natural constituents found in human milk, fish oil or egg yolk. Until recently, infant formulas, though providing the essential fatty acid precursors for these PUFAs, did not contain preformed ARA or DHA. In this study the safety of SUNTGA40S as source of ARA, not only for use in infant formulas but also for nutritional products or food supplements, was evaluated in a subchronic study in Wistar rats, preceded by a 4-week pretreatment period of parental (F(0)) rats and exposure of the F(0) dams throughout mating, gestation and lactation. SUNTGA40S was administered at dietary levels of 0.5%, 1.5% and 5% (wt/wt) adjusted with corn oil to 5.76% added fat. An additional group received 3.65% (wt/wt) SUNTGA40S in conjunction with 2.11% (wt/wt) high DHA Tuna oil, providing an ARA:DHA ratio of 2.7:1. High-fat and low-fat controls received basal diet with or without 5.76% corn-oil supplement. The content, stability and homogeneous distribution of the test substances in the diet were confirmed under study conditions. The administration of SUNTGA40S, with or without DHA oil, did not affect health, growth, fertility or reproductive performance of the parental rats, nor pup characteristics (condition, weight gain, viability, number per litter or sex ratio). In the subchronic study with the offspring (F(1)) rats, no significant differences were found in condition, neurobehavioural observations, ophthalmoscopy, growth, urinalysis or macroscopic and microscopic findings between the test groups and the low-fat or the high-fat controls. In males of the 5% SUNTGA40S and the SUNTGA40S/DHA group, red blood cell counts, haemoglobin concentration and packed cell volume were lower and reticulocytes were slightly higher than in the high-fat and low-fat control groups. Cholesterol, triglycerides and phospholipids in plasma were lower than in the high-fat controls in both sexes in the 5% SUNTGA40S and the SUNTGA40S/DHA group and (for triglycerides only) in the 1.5% SUNTGA group. Due to the administration of extra dietary fat, food intake and prothrombin time (males only) were lower and alkaline phosphatase activity was higher in all the high-fat groups, including the corn-oil controls, as compared to the low-fat controls. The weight of the spleen was higher in males of the 5% SUNTGA40S and the SUNTGA40S/DHA group compared to both the low-fat and the high-fat controls. The effects noted in this study at high dose levels of SUNTGA40S are consistent with previously reported physiological responses to dietary intake of high PUFA containing oils. The present results provide evidence that SUNTGA40S is a safe source of arachidonic acid. Except during lactation when the intake in dams doubled, 5% Suntga40S in the diet was equivalent to an overall intake of approximately 3g/kg body weight/day in F(0) and F(1) animals.

Ultrastructural differences in junctional intercellular communication between highly and weakly metastatic clones derived from rat mammary carcinoma.

We examined by electron microscopy the differences in junctional intercellular communications among highly metastatic clones, weakly metastatic clones, and the parent clone obtained from a spontaneously developed rat mammary carcinoma. We also investigated intercellular communications of the highly and weakly metastatic clone cells with normal fibroblasts. The results showed that ultrastructural changes of the highly metastatic clone cells, such as microvilli, microfilaments, and small organelles including endoplasmic reticulum, Golgi apparatus, and mucous particles, were more distinct than those of the weakly metastatic clone cells, and that the numbers of desmosome and gap junctions of weakly metastatic clone cells were significantly greater than those of highly metastatic clone cells. The formation of gap junctions and desmosomes was found only between weakly metastatic clone cells and normal fibroblasts. When both highly and weakly metastatic clone cells were cultured with normal fibroblasts, a tight junction was observed only in the culture of weakly metastatic clone cells and normal fibroblasts. These results suggest that ultrastructural differences are related to the proliferation and detachment of tumor cells from the primary site in the initial stage of tumor metastasis.

Visualization of cavitated vessels in winter and refilled vessels in spring in diffuse-porous trees by cryo-scanning electron microscopy

Xylem cavitation in winter and recovery from cavitation in the spring were visualized in two species of diffuse-porous trees, Betula platyphylla var. japonica Hara and Salix sachalinensis Fr. Schm., by cryo-scanning electron microscopy after freeze-fixation of living twigs. Water in the vessel lumina of the outer three annual rings of twigs of B. platyphylla var. japonica and of S. sachalinensis gradually disappeared during the period from January to March, an indication that cavitation occurs gradually in these species during the winter. In April, when no leaves had yet expanded, the lumina of most of the vessels of both species were filled with water. Many vessel lumina in twigs of both species were filled with water during the period from the subsequent growth season to the beginning of the next winter. These observations indicate that recovery in spring occurs before the onset of transpiration and that water transport through twigs occurs during the subsequent growing season. We found, moreover, that vessels repeat an annual cycle of winter cavitation and spring recovery from cavitation for several years until irreversible cavitation occurs.

The Progression of Cavitation in Earlywood Vessels of Fraxinus mandshurica var japonica during Freezing and Thawing.

For an examination of the progression of cavitation in large-diameter earlywood vessels of a deciduous ring-porous tree, potted saplings of Fraxinus mandshurica var japonica Maxim. were frozen and then thawed. The changes in the amount and distribution of water in the lumina of the current year's earlywood vessels during the course of the freezing and thawing were visualized by cryo-scanning electron microscopy. When samples were frozen, most of the current year's earlywood vessels were filled with water. After the subsequent thawing, the percentage of cavitated current-year earlywood vessels gradually increased with time. All of the current year's earlywood vessels were cavitated within 24 h, and only limited amounts of water remained in the lumina of earlywood vessels. Similar cavitation of earlywood vessels was observed after thawing of frozen, excised stem pieces. In contrast, many vessels of the current year's latewood retained water in the lumina during freezing and thawing. These observations indicate that the cavitation of the current year's earlywood vessels is not produced during freezing but progresses during rewarming after freezing in F. mandshurica var japonica.

All-solid-state 5-kHz 0.2-TW Ti:sapphire laser system.

We have developed an all-solid-state 5-kHz Ti:sapphire laser system, which produces 22-fs, 0.2-TW pulses. An average power of 22.2 W is the highest ever obtained in ultrashort laser sources. The serious thermal lensing due to high power pumping in a small area of the Ti:sapphire crystal is controlled successfully by a stable quasi-cavity with two concave mirrors.

Vestibular neuropathy accompanying auditory and peripheral neuropathies.

OBJECTIVE: To define the incidence of measurable vestibular disorders in patients with auditory and peripheral neuropathies. DESIGN: Descriptive study of the case features of auditory neuropathy in 14 patients, 8 of whom had concomitant peripheral neuropathies. SETTING: University referral center. PATIENTS: Fourteen patients aged from 10 to 75 years and diagnosed as having auditory neuropathy, 8 of whom had concomitant peripheral neuropathies. MAIN OUTCOME MEASURES: Incidence of abnormal vestibular caloric test results and the relationship of such incidence to clinical variables including the ages of the subjects, the presence of a concomitant peripheral neuropathy, vestibular symptoms, and audiological findings. RESULTS: Abnormal vestibular caloric test results occurred in 9 of the 14 patients. These 9 patients were on average older (35.6 years) than patients with normal caloric responses (17.8 years). Seven of the 9 patients with abnormal caloric responses had concomitant peripheral neuropathies compared with only 1 of the 5 patients with normal caloric responses. None of the 14 patients experienced symptoms of vestibular disorder. CONCLUSIONS: Asymptomatic vestibular disorders are common in patients with auditory neuropathy when a peripheral neuropathy is also present. The reason for the abnormal vestibular test results is likely a neuropathy of the vestibular nerves. Arch Otolaryngol Head Neck Surg. 2000;126:1453-1456

Primary obsessional slowness: long-term findings.

Cases of slowness among patients who spent large amounts of time to perform daily activities were first reported in 1974, and described as primary obsessional slowness (POS). It was observed that it was neither obsessive thoughts nor compulsions that directly hindered their daily activities. However, in more than 20 years following the original report, the diagnostic independence of POS remains controversial, some insisting that obsessional slowness can be explained as secondary. The authors experienced four cases in Japan which share the same characteristics as the original cases. Long-term observation and treatment has led us to support the diagnostic independence of POS. Slowness remained after other accompanying symptoms of obsessive-compulsive disorder had been successfully treated, showing that the slowness of our patients was not secondary.

Application of rapid random stimulation (RRS) to visual evoked potentials in children.

The present study was performed to examine the effects of regular (1 Hz) and modified rapid random stimulation (RRS) (6 and 12 Hz) on visual evoked potentials (VEPs), by simultaneously recording negative waves around 100 ms, wave IV-latency, positive waves around 60 ms, wave III-latency, and amplitudes calculated from peak to peak, without causing impairment of visual acuity, in 44 patients aged 5-17 years. The wave IV-latencies of VEPs obtained by 6 and 12 Hz RRS were easily determined, and the latencies were not significantly changed compared to those obtained by previous 1 Hz regular stimulation. On the other hand, the amplitudes decreased in a frequency-dependent manner (1 < 6 < 12 Hz). These results were found to be similar in both preschool and school children. The examination time of VEPs determined by RRS is one-tenth shorter than that of 1 Hz regular stimulation. Thus, this method has the benefit of shortening the examination time, which decreases fatigue and inattention of the subjects, suggesting that modified RRS is a practically useful method for children.

Seasonal changes in the freezing behavior of xylem ray parenchyma cells in four boreal hardwood species

The freezing behavior of xylem ray parenchyma cells in several boreal hardwood species, namely, Betula platyphylla, Populus canadensis, P. sieboldii, and Salix sachalinensis, was examined by differential thermal analysis (DTA), cryo-scanning electron microscopy (Cryo-SEM), and freeze-fracture replica electron microscopy. Although DTA profiles of samples harvested in summer and in winter suggested that the xylem ray parenchyma cells in all four species responded to freezing stress by extracellular freezing, Cryo-SEM showed clearly that the xylem ray parenchyma cells in all these species responded to freezing stress by shallow supercooling in summer and by extracellular freezing in winter. It is suggested that DTA failed to reveal the true freezing behavior of xylem ray parenchyma cells because of an overlap of temperature ranges between the high-temperature exotherm and the low-temperature exotherm and/or because of the limited extent of the LTE. The seasonal changes in freezing behavior of xylem ray parenchyma cells in all these boreal species, which are results of seasonal cold acclimation, support the hypothesis that a gradual shift of freezing behavior in xylem ray parenchyma cells from shallow supercooling in hardwood species that grow in tropical zones to extracellular freezing in hardwood species that grow in cold areas might be a result of the evolutionary adaptation of hardwood species to cold climates. Copyright 1999 Academic Press.

External and middle ear effects on infant hearing screening test results.

This study investigated the relationship between external and middle ear factors and hearing screening results by automated auditory brain stem response (ABR) and transient-evoked otoacoustic emissions (EOAEs). The ears of 200 healthy newborns aged 5 to 48 hours underwent screening by ABR and EOAE, followed by otoscopic examination. The pass rates for ABR and EOAE were 91% and 58.5%, respectively. On otoscopic examination, 28% (112/400) ears had occluding vernix obscuring the view of the tympanic membrane. Cleaning of vernix was successfully performed in all but 2 ears that had occluding vernix. Cleaning of vernix significantly increased the pass rates of all 400 ears for ABR and EOAE to 96% and 69%. Decreased tympanic membrane mobility was found in 22.7% (90/396) of ears that were evaluated otoscopically. Decreased tympanic membrane mobility had a significant effect on EOAE screening; only 33.4% of ears passed EOAE testing. Decreased tympanic membrane mobility did not significantly affect pass rates for ABR screening; 95% of these ears passed the automated ABR screen. Implications for newborn hearing screening are discussed.

Image analysis of morphological change during arachidonic acid production by Mortierella alpina 1S-4.

The changes in mycelial morphology during arachidonic acid (AA) production by Mortierella alpina 1S-4 were investigated using an image analysis system. Cultivation was performed in a 10-kl fermentor, and the culture broth was separated into two fractions by sieving (0.5 mm aperture size): the filament fraction (F-fraction, <0.5 mm), and the pellet fraction (P-fraction, >0.5 mm). The effect of the mycelial morphology in each fraction on AA production was analyzed. As a result, a product distribution in the culture broth wherein the AA content in the mycelia of the P-fraction was observed to be higher than that in the mycelia of the F-fraction throughout the cultivation. Morphological analysis of the P-fraction revealed that the hairy pellets became smooth because the mycelia on the pellet surface were shaved off; some pellets were broken and reduced in size. The shaved-off mycelia from the hairy pellets surface moved into the F-fraction and aggregated there. From the above findings, it was likely that the low AA content in the F-fraction was due to mycelial damage during the cultivation. In addition, the morphology of the hairy pellets was found to contribute to an increase in the viscosity of culture broth.

Effect of nitrogen source on mycelial morphology and arachidonic acid production in cultures of Mortierella alpina.

The effects of nitrogen source on arachidonic acid (AA) production and morphological changes during the culture of Mortierella alpina were investigated using an image analysis system. When yeast extract, gluten meal, or corn steep liquor was used, a circular pellet morphology was obtained. However, when Pharmamedia, fish meal, or soybean meal was used, M. alpina formed radial filamentous mycelia. The radial filamentous area in the case of soybean meal was 75% of the total mycelial area. In a jar fermentor culture, M. alpina morphology varied with the cultivation period: (i) at 0-6 h culture, the inoculated pellet-like mycelia were adapted to the new environment, (ii) at 6 h-1 d culture, filamentous mycelia grew exponentially which yielded a feather-like morphology, (iii) at 1-2 d culture, the filamentous mycelia became disentangled as a result of the mechanical agitation; consequently, the proportion of filamentous mycelia was increased, (iv) at 2-4 d culture, mycelia showed stationary growth, but the AA concentration increased rapidly, and (v) at 4-6 d culture, hyphae grew thick radially with the AA concentration continuing to increase gradually. In the case of the cultures with feather-like morphology obtained using soybean meal, the AA yield was 0.14 g/g dry cell weight, which was two times higher than that in cultures grown using yeast extract. These results suggest that the feather-like morphology of culture of M. alpina is suitable for AA production.

Neonatal hearing screening with otoscopy, auditory brain stem response, and otoacoustic emissions.

A study was performed to investigate the relationship between external and middle ear factors and hearing screening results by auditory brain stem response (ABR) and transient evoked otoacoustic emissions (EOAEs). The ears of 200 well newborns aged 5 hours to 48 hours underwent screening by ABR and EOAEs, followed by otoscopic examination. The pass rates for ABR and EOAE screening were 88.5% and 79%, respectively. On otoscopic examination, 13% (53 of 400) ears had occluding vernix obscuring the view of the tympanic membrane. Cleaning of vernix was attempted in ears that failed ABR or EOAE screening. Seventeen ears that failed ABR were cleaned, and 12 (71%) of them passed repeat ABR. Thirty-three ears that failed EOAE screening were cleaned, and 22 (67%) of them passed repeat emissions testing. Cleaning vernix increased the pass rates for ABR and EOAE screening to 91.5% and 84%, respectively. Decreased tympanic membrane mobility was found in 9% of ears that could be evaluated otoscopically. Increased failure rates for both ABR and EOAE screening were found in infant ears with decreased tympanic membrane mobility, but significance testing could not be performed because of inadequate sample size. Prevalence of occluding external canal vernix and middle ear effusion as a function of increasing infant age were studied. Implications for newborn hearing screening are discussed.

Effect of consumed carbon to nitrogen ratio of mycelial morphology and arachidonic acid production in cultures of Mortierella alpina.

The influence of the consumed carbon to nitrogen (C/N) ratio on arachidonic acid (AA) production and mycelial morphology was investigated in cultures of Mortierella alpina using shake flasks and a fermentor. The consumed C/N ratio was varied from 5 to 32 under the condition that the total initial amount of carbon and nitrogen sources was 50 g/l. Cellular yield increased markedly at C/N ratios below 7; carbon utilization was switched from cellular growth to lipid biosynthesis in the C/N ratio range of 7-15; lipid biosynthesis was most active when the C/N ratio was in the range of 15-32. However, for C/N ratios higher than 15, the mycelial concentration decreased due to nitrogen limitation but the lipid yield still increased. In the presence of excess nitrogen, the biomass concentration depended on the amount of the nitrogen source, but the AA yield was inversely related to this. On the other hand, in the presence of excess carbon, the fatty acid concentration increased with carbon source concentration but the AA concentration remained constant. From the viewpoint of AA production, the optimum C/N ratio was in the range of 15 to 20 with a balance between the amounts of carbon and nitrogen sources. When an enriched medium was used at a fixed C/N ratio of 20, the cellular and AA concentrations were shown to be proportional to the total concentrations of carbon and nitrogen sources in both flasks and the fermentor. The whole pellet size and width of pellet annular regions did not change with increasing C/N ratio for C/N ratios below 20 in the flask cultures. However, when the C/N ratio was higher than 20, these sizes increased in proportion to the C/N ratio.

99mTc(V)-DMSA and 67Ga-citrate images in a patient with recurrent extraabdominal fibromatosis.

67Ga-citrate and 99mTc(V)-DMSA images were obtained in a patient with recurrent extraabdominal fibromatosis of the left calf. The 67Ga-citrate image showed abnormal uptake in the left calf lesion and right calf, which was considered due to probable myositis caused by overexertion. On the other hand, the 99mTc(V)-DMSA image showed abnormal uptake only in the left calf lesion. Surgical and radiation therapy was performed. Two years after the treatment, two lesions of the recurrent extraabdominal fibromatosis followed, and a 99mTc(V)-DMSA image could demarcate the two lesions. 99mTc(V)-DMSA images were considered to be useful in following up of recurrent extraabdominal fibromatosis.

99mTc-MIBI accumulation in the parathyroid autograft in a patient with recurrent hyperparathyroidism.

A case with recurrent hyperparathyroidism secondary to chronic renal insufficiency is reported. The patient had undergone total parathyroidectomy and autotransplantation of parathyroid tissue five years ago. Bone scintigraphy clearly demonstrated skeletal involvement of secondary hyperparathyroidism and 99mTc-methoxyisobutylisonitrile scintigraphy clearly demonstrated a hyperfunctioning parathyroid autograft.

Scintigraphic evaluation along with CT and MR images in a case of huge gravitation abscess.

The authors presented a rare case of huge gravitation abscess. 67Ga scintigraphy was useful in the evaluation of the extent and activity of the disease. CT showed clearly the location and shape of the abscess. Enhanced MRI showed vertebral lesions as characteristic geographic and ring-like enhancement.

A case of malignant fibrous histiocytoma of the ilium, evaluation by blood-pool scintigraphy.

A rare case of malignant fibrous histiocytoma (MFH) of the ilium was presented and blood-pool scintigraphic images and angiographic images were correlated. Hypervascular tumor and lacking of contrast material with arteriovenous shunts were shown by angiography. Intensive tracer accumulation was shown by blood-pool scintigraphy. After radiation therapy and transcatheter arterial embolization of the tumor, tracer accumulation was noticeable reduced. These findings suggested a vascular tumor such as angiosarcoma, but surgery revealed MFH of the bone. Blood-pool scintigraphy was useful in the evaluation of the vascular characteristics of the tumor.

Newborn hearing screening by otoacoustic emissions and automated auditory brainstem response.

The aim of this study is to compare pass rates for two different hearing screening methods in well newborns as a function of age. Hearing screening tests were performed on 400 ears in 200 healthy newborn infants at the University of california-Irvine Medical Center. The screening methods used were automated auditory brainstem response (ABR) and click evoked otoacoustic emissions (EOAE). The infants' ages ranged from 5 to 120 h, with an average age of 24 h. Overall, 88.5% of ears passed the ABR screen, and 79% passed the EOAE screen. There was no significant difference in the ABR pass rate for infants aged 0-24 h of age as compared with infants aged > 24 h compared with the group aged 0-24 h (P < 0.01). Results are compared with earlier studies and implications for universal hearing screening are discussed.

Comparison of newborn hearing screening by transient otoacoustic emissions and auditory brainstem response using ALGO-2.

The aim of this study is to compare pass rates for two different hearing screening methods in well newborns as a function of age. A previous study by this group compared click evoked otoacoustic emissions (EOAE) and automated auditory brainstem response (ABR) using the ALGO-1 infant hearing screener (Natus Medical, Foster City, CA). Since that study, a new generation automated ABR screener, the ALGO-2, has been developed. In this study, 232 ears in 116 healthy newborn infants aged 5-48 h were tested using the ALGO-2 screener and EOAE. Overall, 92% of ears passed the ABR, while 57% passed the EOAE screen. The ABR pass rate was higher than in the previous study, where 88.5% of ears passed the ABR screen using ALGO-1. The EOAE pass rate in the present study was lower than in the previous study, in which 79% passed the EOAE screen. Pass rates for both EOAE and ABR improved significantly with increasing infant age. There was no significant difference in the test time required for ALGO-2 (5.7 min) compared with EOAE (5.2 min). The results are compared with earlier studies and implications for universal hearing screening are discussed.