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Biomedical subjects

S Asari

Publications and source records attributed to S Asari.

At least 37 records · Page 2Linked to original sources

Bacterial toxins and detection methods.

The diagnosis of infectious disease has been carried out by detecting causative organisms from patient's clinical materials using culture methods so far. However, the method for rapid analysis and diagnosis has been changed by development of advanced methods such as immunological, molecular biological and genetic method. Especially, as a mass outbreak of entrohaemorrhagic E. coli (O157), which is reguired to detect toxins as well as bacteria itself, microbiologists and clinicians recognized the significance of rapid analysis of bacterial toxins. On the diagnosis for the state of disease and decision for the way of the therapy, the rapid detection of bacterial toxin is a significant indispensable tests. From these point of view, introduction of rapid analysis into a laboratory, not using cell culture or experimental animals, is strongly required. Here I describe a various kind of bacterial toxins and their detection methods.

Bacterial Toxins↗

Detection of lipid peroxidation and hydroxyl radicals in brain contusion of rats.

To examine the relationship between the free radicals and brain tissue damage, we investigated the intensity of brain hydroxyl (OH) radical generation and lipid peroxidation in the rat contusion injury model. A unilateral contusion was induced by a weight-drop method. All rats were decapitated six hours after the injury, and brain samples were taken from three portions (core, peripheral, and distal) to examine the specific gravity as an indicator of brain edema, generation of OH using an electron paramagnetic resonance spectrometer (EPR), and malondialdehyde (MDA) and 4-hydroxyalkenals production. Analysis of the specific gravity revealed cerebral edema on the ipsilateral side in the injured group. The signal intensity of EPR in the core and peripheral portions in the contusion group was significantly higher than that in the distal portion of the contusion group and that of all portions in the control animals. No significant difference was observed between the core and peripheral portions of the contusion group. The MDA and 4-hydroxyalkenals production was significantly higher in the core and peripheral portions than in the distal portion of the contusion group and that of all portions of the control group. The degree of posttraumatic brain edema was closely correlated with the increase of DMPO-OH adduct, MDA, and 4-hydroxyalkenals. These results support the current concept that free radical production following traumatic brain injury may induce lipid peroxidation and may be the direct cause of edema formation.

Animals↗

Effects of lecithinized superoxide dismutase on traumatic brain injury in rats.

Only small amounts of superoxide dismutase (SOD) are present in the extracellular space to scavenge excess amounts of superoxide anions (02-) released after traumatic brain injury (TBI). Experiments were performed in rats with cerebral contusion produced by weight-drop technique. We investigated the effects of exogenous lecithinized SOD (PC-SOD) on accumulation of 02- produced in our model, by measuring the level of SOD activity (using the NBT-reducing method) and the expression of copper, zinc-SOD (Cu, Zn-SOD) mRNA (by Northern blot analysis). As determined by tissue-specific gravity, administration of PC-SOD reduced brain edema in the periphery of the lesion 6 h after contusion. SOD activity increased in the peripheral region at 30 min after contusion, but returned to normal levels at 6 h after TBI. Administration of PC-SOD increased SOD activity up to 6 h after TBI. The expression of Cu, Zn-SOD mRNA increased in the core region, peripheral portion, and contralateral hemisphere up to 6 h after TBI, then was suppressed in all three regions by PC-SOD. Our results confirm the important role of 02- in the development of brain edema after TBI and indicate that PC-SOD diminishes brain edema through a protective effect against 02-.

Animals↗

[Evaluation of effective treatment drugs against Acanthamoeba cyst].

Cysts of 2 isolates of Acanthamoeba from the cornea of 2 patients with confirmed Acanthamoeba keratitis were tested in vitro for sensitivity to antimycotic agents such as fluconazole, miconazole, amphotericin-B, pimaricin, antiprotozoal agents such as pentamidine isetionate and antiseptics which could be use in the ophthamological region. Pimaricin was the most successful cysticidal agent against the two strains. Sensitivity to pentamidine isetionate showed variation. Fluconazole, miconazole and amphotericin-B were resistant against cysts with concentration of eye drops that have been used in the treatment of Acanthamoeba keratitis. It was supposed that 5% pimaricin eye drops could be use in the treatment of Acanthamoeba keratitis in addition to keratomycosis. Pentamidine isetionate which belong to the diamidine family, is not yet clear as to the side effects to corneal epithelium cell, but we believe that this drug could be expected as a new therapeutic agent for Acanthamoeba keratitis.

Acanthamoeba↗

Three-dimensional analysis of vasospastic major cerebral arteries in rats with the corrosion cast technique.

BACKGROUND AND PURPOSE: Although mice, rats, and other small animals are commonly used for molecular biology research, their use in the evaluation of cerebral vasospasm after subarachnoid hemorrhage is somewhat problematic because of the correspondingly small size of their cerebral vessels. We have already reported that the corrosion cast technique was useful for evaluating newly formed cerebral vessels in neural grafts in these small animals. In the present study we applied the corrosion cast technique to the evaluation of hemolysate-induced cerebral vasospasm in rats and performed three-dimensional analysis for comparison. The casting was done 10 minutes after the hemolysate injection, so that only acute "vasospasm" was assessed. METHODS: After withdrawal of 0.1 mL cerebrospinal fluid, 0.2 mL hemolysate (n = 9) or saline (n = 10) was injected into the cisterna magna of male Sprague-Dawley rats weighing between 300 and 350 g. Ten minutes later, perfusion of a semipolymerized casting medium was performed at an injection pressure of 100 to 120 mm Hg. The brains were immersed and corroded in 10% NaOH solution. After these procedures, the basilar artery as well as peripheral vessels was analyzed morphologically with scanning electron microscopy. Conventional histological analysis with the use of paraffin-embedded section with hematoxylin-eosin staining was also performed, and the results were compared with those for the corrosion cast methods. RESULTS: In the saline-injected group, SEM showed that the inner surface of the basilar artery was smooth and the form of the endothelial cell was printed on the surface of the cast. In the hemolysate-injected group, the basilar artery showed an apparent vasospasm over its entire length, and corrugation was observed on the inner surface of the basilar artery in a three-dimensional fashion. Higher magnification revealed that the nuclei of the endothelial cells were distorted. Local narrowing of the basilar artery and vasospasm in the arteries of the anterior circulation and in peripheral arteries were also observed. Measurement of the inner diameter of the basilar artery showed 37.8% contraction in the hemolysate-injected group compared with the saline-injected group by the corrosion cast method. This degree of vasospasm was similar to that observed by the conventional histological method. CONCLUSIONS: In this report we show that detailed three-dimensional observation in the rat can be performed qualitatively and quantitatively with the corrosion cast technique. We conclude that this method derives an accurate measurement of the diameter of rat major cerebral arteries and is more reliable for analyzing vasospasm in rats than angiography and other conventional procedures.

Animals↗

[Transient mutism after resection of left frontal lobe astrocytoma in adult: case report].

We reported a case of a 32-year-old female who had a transient mutism after resection of a malignant astrocytoma in the left frontal lobe. Preoperatively magnetic resonance (MR) imaging revealed a cystic tumor in the left frontal lobe. The patient underwent surgery. She was alert but did not speak immediately after surgery. She could follow verbal commands, comprehend written language, and write letters. She had no cranial nerve or extremity paresis. Seven days postoperatively, she began to say simple words, and one month postoperatively she could talk normally. Postoperative MR imaging revealed a hypointensity area on T1 weighted image in the frontal lobe including a part of the anterior cingulate cortex and the anterior part of the corpus callosum. It appears that a dominant hemisphere lesion of both the anterior cingulate cortex and the corpus callosum may be responsible for the development of postoperative mutism.

Adult↗

[Preservation of olfaction in frontal transbasal approach].

Lesions in the frontal base and clival area have conventionally been approached using the transbasal approach described by Derome and Guiot. However, this approach necessitates removal of the crista galli and sectioning of the olfactory rootlets with the associated risk of anosmia and cerebrospinal fluid leak and, in addition, complex reconstruction of the frontal base is required. We describe a new approach to deeply situated tumors in the frontal base, parasellar and clival area which is a modification of Spetzler's craniofacial approach with preservation of olfaction. In this approach, circumferential osteotomy cuts are made around the cribriform plate to permit en block removal with its attachment to both the dura and underlying mucosa. Opening of the dura is avoided and the cribriform bone is used to reconstruct the frontal base. Three patients underwent surgery using this approach for treatment of recurrent pituitary adenoma in two cases and for clivus chordoma in one. In one patient, olfactory function was not preserved because resection of nasal mucosa was small. In the other two patients, however, olfaction was preserved by creating a cribriform plate complex with a sufficient area of resection of nasal mucosa and tumors were completely removed. Olfaction can be preserved, CSF leakage can be prevented, and facial skin incision and complex frontal base reconstruction can be avoided when this technique for maintaining normal olfactory-cribriform anatomy is used in frontal transbasal approaches.

Adenoma↗

[Motor functional MRI by spiral scan--compared with GRE].

Recently, functional MRI (fMRI) has been performed in two methods using 1.5 tesla machine. One was performed using gradient recalled echo (GRE), the other was performed using echo planner imaging (EPI). There is a third method for fMRI called spiral scan. This method is applicable without strong gradient magnetic field such as required by EPI. In this study, motor fMRI was performed in ten normal volunteers by both spiral scan and GRE. Less than 1% risk (t-test), spiral scan showed the reactive area more clearly than GRE. And GRE showed higher signal intensity changing ratio than spiral scan, but there was no significant difference between spiral scan and GRE. The acquisition time of spiral scan was 2 seconds per image, and that of GRE was 10.5 seconds. Spiral scan has good temporal resolution and contrast-noise ratio. These are very advantageous for fMRI.

Adult↗

Proton MR spectroscopy of delayed cerebral radiation in monkeys and humans after brachytherapy.

PURPOSE: To determine whether radiation necrosis can be differentiated from residual/recurrent tumor by proton MR spectroscopy. METHODS: We studied the effects of interstitial brachytherapy on the brains of healthy monkeys and in humans with glioblastoma multiforme. The effects of radiation therapy on normal brain tissue in monkeys were assessed with sequential proton MR spectroscopic studies 1 week to 6 months after brachytherapy. Proton MR spectroscopy was also performed in five patients with residual/recurrent glioblastoma multiforme (three of whom had radiation necrosis after brachytherapy), seven patients with newly diagnosed untreated glioblastoma multiforme, and 16 healthy volunteers, who served as a control group. RESULTS: In monkeys, the ratio of N-acetylaspartate (NAA) to creatine-phosphocreatine (Cr) and the ratio of choline-containing compounds (Cho) to Cr of the reference point were significantly lower 1 week after brachytherapy than before treatment. The ratio of NAA to Cho of the irradiated area tended to be higher 1 week after brachytherapy than before irradiation. These peak metabolic ratios showed characteristic changes 6 months after treatment. In two of three monkeys, lipid signal was elevated 6 months after irradiation. In the clinical study, the ratio of NAA to Cho in the area of radiation necrosis was significantly different from that in glioblastoma multiforme when compared with the contralateral hemisphere after irradiation. In addition, lipid signal was detected in all patients with radiation necrosis. CONCLUSION: It might be possible to use proton MR spectroscopy to differentiate radiation necrosis from residual/recurrent glioblastoma multiforme on the basis of comparisons with the contralateral hemisphere after radiation therapy.

Adult↗

The relationship between brain surface elastance and brain reexpansion after evacuation of chronic subdural hematoma.

BACKGROUND: Brain stiffness has been thought to be a factor affecting brain re-expansion after the evacuation of chronic subdural hematoma. METHODS: As an index of brain stiffness, the pressure for compressing the brain using an ophthalmodynamometer was defined as brain-surface elastance. This elastance in 14 patients was measured and analyzed in relation to brain re-expansion. RESULTS: Patients with an enlarged subdural space on computed tomography 1 month after the operation had higher elastance. There was a correlation between high elastance and high age. CONCLUSIONS: Measuring the elastance after evacuation of the hematoma may help predict the persistence of a subdural space.

Aged↗

Role of extracellular matrix in experimental vasospasm. Inhibitory effect of antisense oligonucleotide on collagen induction.

BACKGROUND AND PURPOSE: Although it has been suggested that collagen plays a role in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage, there has been no constructive research to prove it directly. In this study we stopped the transcription of the procollagen type I gene by introducing antisense oligonucleotides for its mRNA in a rat femoral artery model of vasospasm induced by blood and assayed the changes in the vasoconstrictive activity of the vessel and expression of the procollagen mRNA. METHODS: We applied antisense, sense, or missense oligonucleotides, located at the carboxyl propeptide region for alpha 1(I) procollagen mRNA, onto the femoral artery in a rat femoral artery model of vasospasm. The diameter of the artery was measured by angiography. The transcription level of the procollagen gene in the arterial tissue was assayed by use of reverse transcription-polymerase chain reaction. Morphological change in the artery was observed with aldehyde-fuchsin-Masson-Goldner staining. RESULTS: In the model, when the artery was exposed to antisense oligonucleotides in pluronic gel for 5 days to prevent arterial contraction, the contraction was inhibited at a significant level (76.0% +/- 5.6) when compared with that in control experiments using sense oligonucleotides (64.0% +/- 2.4), missense oligonucleotides (63.5% +/- 3.5), or gel alone (62.1% +/- 5.8). The application of antisense oligonucleotide resulted in a marked decrease in alpha 1(I) procollagen mRNA expression as determined by polymerase chain reaction, indicating that the collagen reduction by antisense oligonucleotides occurred at the transcription level. Histological staining suggested that collagen accumulation at the site in the artery where antisense oligonucleotide had been administered was indeed less than that in the control artery. CONCLUSIONS: The results indicate that the induction of procollagen type 1 could cause pathogenesis of the arterial contraction induced by blood in a rat femoral vasospasm model.

Animals↗

Chromaffin cell survival and host dopaminergic fiber recovery in a patient with Parkinson's disease treated by cografts of adrenal medulla and pretransected peripheral nerve. Case report.

A 55-year-old woman with severe Parkinson's disease was treated by cografting adrenal medulla with pretransected peripheral nerve into the bilateral caudate nuclei. The patient showed modest improvement of her akinesia; this effect persisted for 1 year after transplantation, when she suddenly died from upper gastrointestinal bleeding unrelated to the grafting procedure. At autopsy, a large number of tyrosine hydroxylase-immunoreactive chromaffin cells were observed within the caudate graft sites and a dense network of host dopaminergic fibers was visualized. This autopsy finding is very important for the field of experimental and clinical chromaffin cell grafting because it is the first evidence that cografts using pretransected peripheral nerve might enhance the survival of chromaffin cells and the recovery of host dopaminergic fibers in humans suffering from Parkinson's disease.

Adrenal Medulla↗

Inhibition of vascular contraction by intracisternal administration of preproendothelin-1 mRNA antisense oligoDNA in a rat experimental vasospasm model.

To clarify the role of endothelin-1 (ET-1) in the etiology of hemolysate-induced contraction of vessels, the authors introduced antisense oligoDNA for preproendothelin-1 (ppET-1) messenger RNA in a rat model of vasospasm. Phosphorothioate antisense oligoDNAs for ppET-1 were injected into the cisterna magna. Fluorescein isothiocyanate-labeled phosphorothioate antisense oligoDNAs were proven by fluorescence chasing to be incorporated into the vascular wall. Striking inhibitory effects of experimental vasospasm were observed in the basilar artery (BA) in which the oligoDNAs were injected. The vascular contraction was significantly inhibited by oligoDNAs after 20 minutes of hemolysate exposure, which suggested that ET synthesis started approximately 20 minutes after hemolysate stimulation. Expression of ppET-1 in the BA in which the spasm was inhibited was markedly suppressed at the transcription level. The results indicate that ET-1 may play an important role in hemolysate-induced vasoconstriction in rats. In addition, the antisense approach in the cerebrospinal fluid might be a useful tool for preventing cerebral vasospasm.

Animals↗

[Encapsulated dopamine-secreting cells transplanted into the brain: a possible therapy for Parkinson's disease].

Encapsulation of neurosecretory cells within a semipermeable membrane may possibly isolate the enclosed cells from the host immune system and allow inward diffusion of nutrients and outward diffusion of neurotransmitters. Moreover, the encapsulation procedure may prevent the tumor formation of enclosed cells, when they are derived from tumor cells. In the present study, PC12 cells, a dopaminergic cell line derived from a rat pheochromocytoma, were enclosed within an agarose/poly (styrene sulfonic acid) (agarose/PSSa) mixture and transplanted into the brains of rats (allogeneic transplantation) or guinea pigs (xenogeneic transplantation). Tyrosine hydroxylase (TH) immunoreactive PC12 cells within the microcapsules were observed in all rats and guinea pigs at least up to five weeks after transplantation. PC12 cells were round in shape and of relatively uniform small size. Although PC12 cells occasionally formed cell clusters, the formation of a tumor was not observed. The host reaction to agarose/PSSa microcapsules was minimum. The degree of glial fibrillary acidic protein (GFAP) positive astrocyte density around the microcapsules was similar to that around injection tracks. There was no apparent immunological rejection around the capsules. High-performance liquid chromatography with electrochemical detection (HPLC-EC) showed basal and potassium-evoked release of dopamine from the PC12 cell-enclosed microcapsules in vitro. Although our data is preliminary, we believe that agarose/PSSa microcapsules are promising for producing semipermeable membranes that enable allo-and xenotransplantation of neurosecretory cells into the brain in the absence of systemic immunosuppression. This approach is expected to be applied in Parkinson's disease in the near future.

Adrenal Gland Neoplasms↗

Expression of Cu,Zn-superoxide dismutase mRNA after cold and contusion injury in the rat brain.

We analyzed the expression of Cu,Zn-superoxide dismutase (SOD) mRNA in both contusion and cold injury. Twenty-three rats were divided into 3 groups: a control group, a contusion group, and a cold injury group. Six hours after the injury, the rats were decapitated and the gray matter was resected from 3 portions: the core of the injured cortex, its periphery, and a distal portion on the non-lesion side. Based on the specific gravity of each sample, almost the same degree of edema developed in both injury groups. The mRNA expression in the cold injury group, however, significantly decreased in all portions. The extremely low temperature associated with cold injury is a possible cause of the decrease in Cu,Zn-SOD mRNA.

Animals↗