Isolation of novel mycobacteria contaminating an aquarium fish tank in a Japanese university hospital.

AIMS: To better understand nontuberculous mycobacteria (NTM) contamination in a hospital setting, six freshwater fish gut homogenates and water in an aquarium fish tank placed on the reception counter of a nursing station were cultured for mycobacteria. METHODS AND RESULTS: By direct sequencing of 16s rRNA, rpoB and hsp65, scotochromogenic and nonchromogenic Mycobacterium szulgai isolates containing hsp65 type II (GenBank accession nos. FJ384762 and FJ384764, respectively), Mycobacterium gordonae isolates containing rpoB clusters B and E (GenBank accession no. FJ384766), and Mycobacterium kansasii isolates containing hsp65 type VI were collected from the gut homogenates and water from the fish tank. However, no isolates were obtained from the tap water used to refill the fish tank. A randomly amplified polymorphic DNA (RAPD) analysis using a 10-mer primer (5'-TGGTCGCGGC) showed that some NTM from the fish tank water were identical to those obtained from the gut homogenates. CONCLUSIONS: Fish and water in the tank were contaminated by the novel NTM. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings could help to elucidate infection routes and contamination sources of novel NTM from water sources.

A 12 month in vivo study on the response of bone to a hydroxyapatite-polymethylmethacrylate cranioplasty composite.

We investigated the osteoconductivity and biocompatibility in vivo of a new hydroxyapatite-polymethylmethacrylate (HA-PMMA) composite developed for use as an implant material for cranioplasty, which is expected to have the good osteoconductivity of HA together with the strength and ease of handling of PMMA. The HA-PMMA composites were implanted in eight full-grown beagles and then 6, 12, 24 weeks and 1 year after implantation, the animals were sacrificed and the implanted materials removed along with the surrounding tissues. Extirpated specimens were studied using an optical microscope and micro-computed tomography (micro-CT). Fibrous connective tissue was prominent in the interface of the composite at 6 weeks. New bone formation was seen around the implant, 12 and 24 weeks after operation. At 1 year, new bone filled in the interface of the HA-PMMA composite and adhered to the surrounding autogenous bone. Mixing HA and PMMA did not interfere with the osteoconductivity of the HA component. In micro-CT findings, the new bone growing on the HA-PMMA composite could be seen attaching preferentially to HA particles exposed at the composite surface, rather than the PMMA. This study demonstrated that this HA-PMMA composite is a good candidate for cranial bone implants due to its good osteoconductivity and biocompatibility.

Contaminated oral intubation equipment associated with an outbreak of carbapenem-resistant Pseudomonas in an intensive care unit.

Twenty intensive care patients were diagnosed as infected or colonized by Pseudomonas aeruginosa within a one-month period; a rate three to four times higher than the typical background frequency of this infection in the intensive care unit (ICU). Patients with positive respiratory specimens were mechanically ventilated, which included re-used disinfected bite blocks during intubation. Fourteen specimens from 20 positive patients originated in the respiratory tract. Seven clonal variants were isolated and identified as originating from the same strain by pulsed-field analysis. These isolates were also matched to the strain detected on the re-used bite blocks, which had been disinfected with 140ppm sodium hydrochloride. Notably, Staphylococcus aureus was also detected on bite blocks sterilized with ethylene dioxide, indicating incomplete disinfection. In immunocompromised patients, re-use of bite blocks during intubation must be prohibited. Single-use kits or intubation without the use of bite blocks is recommended.

Astrocytes co-express aquaporin-1, -4, and vascular endothelial growth factor in brain edema tissue associated with brain contusion.

INTRODUCTION: Brain edema may be life threatening. The mechanisms underlying the development of traumatic brain edema are still unclear; however, mixed mechanisms including vasogenic, ischemic, and neurotoxic types of edema may be contributors. Recent studies indicate that astrocytes, aquaporins (AQPs; a protein family of water channels), and vascular endothelial growth factor (VEGF) may have important roles in the formation and resolution of brain edema. We studied the expression of AQPs and VEGF in the edematous brain. METHODS: We investigated the expression of AQP1, AQP4, and vascular endothelial growth factor (VEGF) in contusional brain tissue surgically obtained from 6 patients. Glial fibrillary acidic protein (GFAP) was also stained to detect astrocytes and to clarify the location of those proteins. The specimens received immunohistological staining and 3-color immunofluorescent staining, and were observed using confocal laser scanning microscopy. RESULTS: AQP1, AQP4, and VEGF were co-expressed in GFAP-positive astrocytes. AQP1 and AQP4 were expressed strongly in astrocytic end-feet. The astrocytes were located in the edematous tissue, and some cells surrounded cerebral capillaries. CONCLUSION: Our results suggest that AQP1, AQP4, and VEGF are induced in astrocytes located in and surrounding edematous tissue. Those astrocytes may regulate the water in- and out-flow in the injured tissue.

Assessment of postischemic cerebral energy metabolism in cat by 31P NMR: the cumulative effects of secondary hypoxia and ischemia.

The sensitivity of cerebral energy metabolism to ischemic and hypoxic stresses following global cerebral ischemia was evaluated in a cat model using 31P nuclear magnetic resonance (NMR) spectroscopic methods. Complete global cerebral ischemia of 5 to 10 min in length was produced at 1 h intervals by reversible arterial occlusion, permitting continuous monitoring of NMR and EEG. Ischemia appeared to produce slightly more severe energy failure in animals that had previously experienced an ischemic injury. Preischemic hypoxia (5% O2 for 5 min) resulted in minor changes in the levels of phosphocreatine and intracellular inorganic phosphate, which were slightly amplified in animals that previously experienced ischemia.

Global cerebral ischemia associated with cardiac arrest in the rat: I. Dynamics of early neuronal changes.

Light microscopic neuronal changes were studied in rats subjected to 10 min of global ischemia produced by compression of the major cardiac vessels. Observations of cresyl violet-stained sections revealed early changes involving predominantly GABAergic neurons in various locations. In rats killed 15 min after recirculation, the changes were characterized by the appearance of a clear peripheral zone with condensation of the remaining neuronal cytoplasm. After 1 h, these zones appeared to be compartmentalized into individual pearl-like vacuoles, especially prominent in the nucleus reticularis thalami. After 3 h, the cytoplasmic vacuoles disappeared and the neuronal changes, particularly in the cerebral cortex, striatum, hippocampus, and pars reticulata of the substantia nigra, consisted mainly of hyperchromasia or loss of Nissl substance. After 2 days, the cerebral cortex and thalamus contained occasional neurons with conspicuously large nucleoli. After 7 days, the hippocampus revealed an approximately 50% loss of CA1 pyramidal neurons, associated with intense microglial reactivity in the stratum radiatum, whereas the neuronal destruction was more complete in the nucleus reticularis thalami. Our observations suggest a possibility that early changes in GABAergic neurons may provide a period of neuronal disinhibition and thus contribute to an excitatory ischemic damage in regions connected by GABAergic circuitry.

Immunohistochemical detection of progesterone receptors and the correlation with Ki-67 labeling indices in paraffin-embedded sections of meningiomas.

Female sex steroids may play a role in the proliferation of meningiomas. We investigated the progesterone receptor (PgR) immunoreactivities and the Ki-67 labeling indices in the formalin-fixed, paraffin-embedded sections of meningiomas from 39 patients. After autoclave pretreatment of the sections (which were immersed in a citrate buffer), the sections were incubated with the monoclonal antibody for the PgR and the MIB-1 monoclonal antibody for the Ki-67 antigen. In the meningiomas studied, the immunoreactivity for the PgR was moderately to strongly positive in 51%, weakly positive in 21%, and negative in 28%. The nuclear staining for the PgR was clear, and no tumors were positive for the estrogen receptor. The Ki-67 labeling indices of the PgR-positive meningiomas (mean +/- standard deviation, 2.35 +/- 2.12%) were significantly lower than those of the PgR-negative meningiomas (6.53 +/- 4.83%) (P < 0.05). Two meningiomas that had recurred more than once showed high Ki-67 labeling indices and negative immunostaining for the PgR. These findings indicate that the PgR status may be closely related to the growth potentials of the meningiomas. Our results confirm that the immunodetection of the PgR and the Ki-67 antigen on the paraffin sections of meningiomas provides a practical tool for estimating the biological behavior of the meningiomas.

Different distribution of c-myc and MIB-1 positive cells in malignant meningiomas with reference to TGFs, PDGF, and PgR expression.

We investigated the expression of transforming growth factors (TGFs), platelet-derived growth factor (PDGF), progesterone receptor (PgR), and c-myc in 20 cases of meningioma of various grades: 17 benign, 2 atypical, and 1 anaplastic. All cases of atypical and anaplastic meningioma were positive for c-myc, whereas all 17 benign meningiomas were negative for c-myc immunostaining. Expression of TGF-alpha, TGF-beta, and PDGF-BB proteins was seen in more than 80% of the meningioma cases and was not restricted to their histological grade of meningioma. PgR was expressed mainly in benign meningiomas. Moreover, the cells expressing c-myc protein were not usually stained by MIB-1. These results indicate that c-myc does not directly work on the proliferation of meningioma cells, and even in homogeneous meningioma cells, there may be many functional variations that lead the meningioma cells to their growth.

Repetitive ischaemia of cat brain: pathophysiological observations.

The cumulative effect of repetitive ischaemia on brain injury was studied in halothane-anaesthetized cats submitted to three episodes of global cerebro-circulatory arrest. Ischaemia of 5.0, 7.5 and 10.0 min duration was produced at hourly intervals by intrathoracic clamping of the innominate and subclavian arteries, and the resulting pathophysiological changes were evaluated by recordings on the electroencephalogram (EEG), blood flow and specific gravity. During each episode of ischaemia EEG flattened within 15 s. After ischaemia the latency of EEG recovery increased with the duration and with the number of repetitions of each ischaemic episode, indicating cumulation of electrophysiological impairment. The flow studies revealed a minor degree of hyperaemia after each ischaemic episode, followed by severe hypoperfusion in the caudate nucleus but not in the cerebral cortex. Brain oedema - as assessed by specific gravity measurements - developed in the hippocampus after three episodes of 5 min ischaemia, and in all grey matter structures investigated after three episodes of 10 min ischaemia. To evaluate the resistance of the ischaemically injured brain to respiratory hypoxia, total oxygen was repeatedly reduced to 5% for 5 min. During these episodes EEG activity progressively declined as a function of the length and the repetition of ischaemia. Parallel n.m.r. spectroscopic measurements in the same model have demonstrated that disturbances of brain energy state during the hypoxic episodes are minor even after three episodes of 10 min ischaemia. EEG suppression, in consequence, is an electrical shut-down phenomenon for the maintenance of cerebral energy state under critical conditions of oxygen delivery.

Cardiac arrest-induced complete cerebral ischaemia in the rat: dynamics of postischaemic in vivo calcium uptake and protein synthesis.

Dynamics of pathological changes following cardiac arrest induced cerebral ischaemia were related to the findings of double tracer autoradiography of 45Ca and 3H leucine uptake as respective indicators of ischaemic injury and metabolic disturbance. Abnormal calcium accumulation, determined by 45Ca uptake, was related to injured but still living neurons and to reactive glial elements. 45Ca autoradiography confirmed a high sensitivity to neuronal injury of the nucleus reticularis thalami (NRT), hippocampal CA1 pyramidal layer, inferior colliculus, ventral thalamic nucleus (VTN), caudate nucleus, and parietal cortex. 3H leucine incorporation revealed that an initially widespread inhibition of protein synthesis was followed by its considerable recovery. Observations concerning hippocampal CA1 sector and VTN suggested that a significant degree of protein synthesis, maintained at the late stage after postischaemic recovery, was related to survival and regeneration of neurons and not to the presence of glial elements.

Involvement of disregulated c-myc but not c-sis/PDGF in atypical and anaplastic meningiomas.

We investigated the expression of c-myc and c-sis/PDGF mRNA and protein products in 20 cases of meningiomas of various grades: 10 benign, 5 atypical and 5 anaplastic meningiomas. All cases of atypical and anaplastic meningiomas were positive for c-myc protein and mRNA by immunohistochemistry and in situ hybridisation, respectively, while all 10 benign meningiomas were negative for c-myc immunostaining, with only one benign tumour positive for c-myc mRNA. Expression of PDGF-BB protein and c-sis mRNA were seen in more than 80% of the meningioma cases and was not restricted to the histological grades of meningiomas. Semiquantitative analysis showed that the frequency of c-myc immunopositive cells positively correlated with Ki-67 proliferative indices. Our findings suggest that c-myc, but not c-sis/PDGF, has some concern to the malignancy of meningiomas.

Fluorescence automatic cell sorter and immunohistochemical investigation of CD68-positive cells in meningioma.

Infiltration of brain neoplasms by mononuclear cells including monocytes/macrophages has attracted little attention since they have marked morphological heterogeneity. Twenty-seven meningiomas were studied by anti-CD68 antibody-gated flow cytometry and by immunohistochemical analysis using the anti-CD68 antibodies. Flow cytometric analysis divided cells contained within tumor tissues into CD68-positive and -negative cells. In addition, eight gliomas, eight metastatic brain tumor, and 12 pituitary adenomas were investigated in the same way to compare meningiomas. The mean contents of CD68-positive cells were 24.0 +/- 3.7% in meningiomas, 4.4 +/- 1.4% in gliomas, 9.5 +/- 3.9% in metastatic brain tumors, and 4.5 +/- 1.8% in pituitary adenomas. Immunohistochemically, CD68-positive cells showed significant heterogeneity and were detected as round, rod-shaped, ameboid and ramified cells in meningiomas. Although the infiltrated mononuclear cells in gliomas have been investigated to some degree and showed that they express cytokines and/or growth factors, these infiltrated cells in meningioma have barely been studied. The CD68-positive cells detected in this study are likely to be monocytes, macrophages and microglias, and are presumed to be in various functional stages and to play important roles in growth regulation in meningioma.

Recurrence in meningeal hemangiopericytomas.

BACKGROUND: Meningeal hemangiopericytomas are more aggressive than typical meningiomas, with a high rate of recurrence and distant metastases. The question of whether a correlation exists between prognosis and histologic features remains controversial. CASE DESCRIPTION: We report two cases of recurrent meningeal hemangiopericytomas. Although local growth control of the tumor was obtained by tumor removal and irradiation in a 38-year-old male patient (Case 1) with a recurrent tentorial tumor, the tumor disseminated and metastasized extracranially within a short period after treatment, leading to rapid deterioration. Another 38-year-old female patient (Case 2) with a recurrent orbital tumor had a favorable outcome after tumor removal. The Ki-67 proliferative index using the MIB-1 monoclonal antibody increased as the tumor recurred in Case 1 (2.5%, 7.9%, and 15.7%), but did not change between primary and recurrent tumors of Case 2 (4.2%, 3.1%). Immunostaining for p53 protein in Case 1 was negative at the first resection, and became positive at the second and third resections, whereas in Case 2, it was negative in both the primary and recurrent tumors. CONCLUSIONS: Our results suggest that p53 protein accumulation with a high proliferative potential is a useful marker to estimate malignant progression in meningeal hemangiopericytomas.

Graphic analysis of microscopic tumor cell infiltration, proliferative potential, and vascular endothelial growth factor expression in an autopsy brain with glioblastoma.

BACKGROUND: Growth of brain tumors requires tumor-cell attachment to adjacent structures, degradation of surrounding matrixes, migration of tumor cells, proliferation of vasculature, and tumor cell proliferation. Comparison of the findings on neuroimaging, degrees and patterns of tumor invasion, regional tumor cell viability detected by Ki-67 immunohistochemistry, and regional vascular endothelial growth factor (VEGF) expression in whole-brain specimen of glioblastoma therefore is of great interest, and will facilitate study of the host reaction against the glioblastoma. METHODS: We graphically analyzed microscopic tumor-cell infiltration, regional differences in Ki-67 labeling indices (LI), and immunohistochemical expression of VEGF in an autopsy brain with glioblastoma. RESULTS: Glioblastoma cells infiltrated the brain far beyond the gross limits of the tumor and the areas with high signal intensity on T2-weighted magnetic resonance images. A wide range of histologic malignancy was apparent from hematoxylin-eosin staining and the Ki-67 labeling indices. VEGF was highly expressed in normal astrocytes located outside the tumor. CONCLUSION: Graphic analysis of histologic and immunohistochemical patterns is a useful method of investigating the mechanisms of glioma growth, tumor cell infiltration in the brain, and the host reaction of the brain against neoplasms.

Tissue reconstruction process in the area of peri-tumoural oedema caused by glioblastoma--immunohistochemical and graphical analysis using brain obtained at autopsy.

BACKGROUND: In the area of peri-tumoural oedema, proteolytic agents derived from the tumour cause tissue degradation, which promotes tumour cell invasion. METHOD: We investigated the biological processes in the area of peri-tumoural oedema, using a brain obtained at autopsy from a patient who died from glioblastoma. Immunohistochemistry was performed to detect vascular endothelial growth factor (VEGF), c-myc, p53, paternally expressed gene-3 (PEG-3), transforming growth factor beta (TGFB), and tumour necrosis factor alpha (TNFA). The data were translated into colour graphics and the localization of these proteins was analyzed. FINDINGS: In the area of peri-tumoural oedema, Ki-67 and p53 positive cells were observed with TGFB expression. Moreover, c-myc, PEG-3, VEGF, and TNFA were also expressed strongly in the glial cells or extra-cellular spaces in the area of peri-tumoural oedema. INTERPRETATION: These data suggest that in the area of peri-tumoural oedema, tissue reconstruction processes take place with concomitant anti-tumour activities. The expression of c-myc, VEGF, and TNFA in the area of peri-tumoural oedema may indicate that these proteins are not utilized for tumour growth, but may be used to guard the brain against tumour invasion. Peri-tumoural oedema does not only indicate the tissue damage caused by tumour, but many tissue reconstruction processes take place in these areas against tumour cell invasion.

Correlation of magnetic resonance imaging and histological findings in a large basilar tip aneurysm after coil embolization--case report.

A 73-year-old female with a large basilar tip aneurysm underwent endovascular coil embolization with interlocking detachable coils (IDCs). The patient subsequently died of pneumonia 25 days after the embolization, and the aneurysm specimen was obtained at autopsy. Histological examination showed that the intraaneurysmal structure consisted of three layers. The outer layer was mildly organized thrombus surrounding the coils, the middle layer was disorganized clot, and the inner layer consisted of fresh blood clot. Gradient-echo magnetic resonance (MR) imaging taken before death had demonstrated a central region of high intensity and a peripheral low intensity region corresponding to the inner and middle-outer layers of the aneurysm, respectively. Intraaneurysmal placement of IDCs leads to the formation of a clot surrounding the coils. However, clot formation may be inadequate where the packing of the IDCs is incomplete. Comparison of the MR imaging and histological findings in this case show that gradient-echo MR imaging can assess thrombus and residual blood flow within the aneurysm.

[Avascular necrosis of the femoral head caused by steroid treatment in neurosurgery].

Steroid induced avascular necrosis of the femoral head is a well known disease, but, there are few reports about the disease in neurosurgical patients. In the neurosurgical field, the use of steroids has become prevalent since the 1960's. Recently, the adverse effect of steroids and the limitation of its effect have been highlighted, but its use against neurosurgical diseases is still a common treatment to prevent cerebral edema or to counteract hypo-pituitarism caused by hypophyseal lesions. We reviewed 250 patients of avascular necrosis treated between 1985 and 1997 in our institute. Within these patients, 11 (4.4%) were treated with steroid during neurosurgical treatment. Six patients were treated for brain tumors near hypophyseal lesions, and 5 patients were treated for head injury or cerebro-vascular disease. It is concluded that total steroid dose over 5000 mg such as hydrocortisone may become a high risk for causing avascular necrosis of the femoral had in neurosurgical disease, and it may occur even with the supplemental steroid treatment against hypo-pituitarism. The onset is usually 2 or 3 years after the neurosurgical treatment, when neurosurgical care is no longer needed. Therefore, it tends to be ignored in the neurosurgical field. The treatments against avascular necrosis of the femoral head were femoral head osteotomy or conservative management, and good results were obtained. Early diagnosis and early treatment is essential. Further consideration concerning steroid treatment in neurosurgical patients may be required.

[Changes in QOL in patients with brain tumors measured by mood changes during and after treatment].

UNLABELLED: Patients with brain tumors are exposed to severe stress which may have an influence on their quality of life (QOL). To measure QOL in those patients, we measure their mood state during and after the treatments. MATERIALS: 16 patients who were admitted to our department for treatment of brain tumors, were included in the study. The tumors included 5 gliomas, 6 meningiomas and others. They were 7 males and 9 females, and age distributed from 19 to 74 years old. All patients presented more than 90% of the Karnofsky performance score (KPS) on discharge, so they were expected to return to their previous social life. METHOD: The self-answering tests were performed on admission (Pre), on discharge (Post 1), and at more than 5 months after the discharge (Post 2). The tests included an original questionnaire asking consent to admission and treatment, and concerning the feeling of disability to cope with conditions of living, Cornell Medical Index (CMI) which measured the neurosis, and Profile of mood state (POMS) which measured the 6 subscales of patients mood states including tension-anxiety (TA), depression (D), anger-hostility (AH), vigor (V), fatigue (F), and confusion (C). RESULTS: The questionnaire showed that the patients feel satisfied with having consented to the treatment. The feeling of disability to cope with living became stronger in Post 2 than in Pre. CMI showed a borderline of neurosis in three patients on admission, and in four patients in Post 2. Only the TA of POMS subscale improved significantly in Post 2. However, other subscales were unchanged. It is characteristic that all of the subscales of POMS showed less disturbance on discharge compared with that on admission, but, they returned to the Prelevel after they returned to social life. CONCLUSIONS: Patients with brain tumors have satisfactory consent to the treatment, however, they feel disability to cope with social life. On discharge, they showed a better mood state compared to that on admission, but the mood state turned for the worse again during the follow-up period. It is evident that patients with brain tumors are exposed to severe stress even after the completion of the treatment. The results necessitate our taking patients' mental health into consideration for our treatment protocol.

[Irinotecan (topoisomerase-I inhibitor) for the treatment of recurrent primary intracranial malignant lymphoma].

Primary intracranial malignant lymphoma is a fetal disease with poor prognosis, and there is no effective treatment against recurrent primary intracranial malignant lymphomas. We report 3 cases of malignant lymphoma treated with irinotecan (topoisomerase-I inhibitor, camptothecin derivatives), an aromatic drug extracted from camptotheca acuminata. After the initial diagnosis, surgical resection followed by radiation therapy was performed for one cerebral, and two cerebellar malignant lymphomas. The tumors recurred 1 month, 18 months, and 18 months after the initial treatment, respectively. The former two cases were treated with additional radiation therapy and/or radiosurgery for the recurrent tumors; however, the tumors recurred again. All cases were treated finally with a combination therapy of irinotecan and cis-platinum followed by a maintenance therapy with irinotecan only. All cases showed a sharp roentgenographical response to the chemotherapy even after cumulative recurrences. One patient died of systemic infection, and another died of intracranial tumor recurrence 11 and 29 months after the initial diagnosis, respectively. Autopsies revealed multiple tumor recurrences in both these cases. The other patient died 31 months after the initial diagnosis, also due to intracranial tumor recurrences. These results indicate the usefulness of irinotecan for the treatment of recurrent primary intracranial malignant lymphoma; however, further investigation is necessary to establish a better protocol for irinotecan treatment against primary intracranial malignant lymphoma.

Cumulative effect of repetitive ischemia: pathophysiological findings.

The cumulative effect of ischemia on the brain was investigated in cats using a repetitive transient global ischemia model. The cats were submitted to three series of repetitive ischemia of 5.0, 7.5 and 10.0-minute durations at 1-hour intervals by intrathoracic clamping of the innominate and subclavian arteries. Pathophysiological changes during and after the ischemic episodes were evaluated by monitoring the electroencephalograms (EEG), cerebral blood flow (CBF), specific gravity and 31P-MR spectroscopy (MRS). Transient 5.0, 7.5, and 10.0-minute ischemias appeared to produce a slightly more severe energy failure on the 31P MRS measurement in the animals that had previously experienced an ischemic injury than those that had not. Additionally, repetition of ischemic episodes at 1-hour intervals led to a progressive lengthening of the duration of the spontaneous electrocortical suppression that followed each ischemic episode. However, preischemic hypoxia (5% O2 for 5 minutes) resulted in minor changes in the levels of phosphocreatine and intracellular inorganic phosphate on the MRS measurement, otherwise the EEG activity declined progressively. This shut-down response of the EEG can be concluded to serve in preserving the energy state of the brain although it is not capable of preventing the development of postischemic brain edema and neuronal death.