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The purpose of our research is to identify causative metals by using particle-induced X-ray emission (PIXE) method to directly analyze trace elements in the oral mucosa with oral lichen planus (OLP). Subjects were 72 patients with OLP, and the patients are 25 men and 47 women, with a mean age of 60.1 years (OLP group). The control is elemental analysis data by PIXE method of 100 cases of the oral mucosa of healthy persons (control group). Seventeen essential elements and 12 contaminating elements were detected from the oral mucosa of the OLP and control groups. As to appearance ratio of elements in oral mucosa, among the contaminating elements, levels of Al, Ga, Sb, Hg and Pb were significantly lower, whereas Au and Y were significantly higher in the OLP group than in the control group. About concentration of elements in oral mucosa, the contaminating elements in the oral mucosa of the OLP group had a lower appearance ratio than in the control but each of the contaminating elements was more abundant. Among 25 subjects in the OLP group and seven in the control group, serum, oral mucosa and saliva were collected from the same individual. Abundance in serum and saliva varied depending on the element, but in general, a tendency was seen for contaminating elements to be more abundant in saliva than in serum and for essential elements to be more abundant in serum than in saliva.

The effects of the vascular disrupting agent AVE8062 on tumor and normal tissue samples were investigated by particle-induced X-ray emission (PIXE) analysis using quantum dots (QDs). We investigated fibrosarcoma tumors in mice, and used kidney tissue as a control. Non-targeted QDs were used to characterize the tissue regions where blood flow is interrupted by AVE8062. We found that the concentration of the QDs in the tumors and kidneys exposed to AVE8062 was lower than that of the control group. Sub-millimeter PIXE analysis (with a beam size of 0.5 [Formula: see text] 0.5 mm2) was used to investigate the spatial distribution of QDs in the tissue samples. We found that the QDs were accumulated in localized regions of the kidney section of the AVE8062-treated group whereas the QDs were uniformly distributed in the control kidney. This suggests that AVE8062 caused blood flow interruption not only in the tumor samples but also in the normal blood vessels in the kidneys.

The aim of this study was to determine whether oxygen-releasing microcapsules could be used to sensitize cancer cells to kill by radiation. The microcapsules were generated by spraying a mixture of 0.1% alginate and hyaluronic acid into a 0.3 mmol/l solution of CaCl2 and FeCl2. These were then subcutaneously injected around a MM48 tumor (a cell line derived from human breast cancer) in the left hind legs of C3H/HeN mice, and tumors were dosed with 60Co γ-ray radiation. We showed that the oxygen released from the microcapsules enhanced the anti-tumor effect of radiation treatment via the generation of oxygen radicals.

The aim of this study was to evaluate the reliability and effectiveness of direct determination of trace and major element concentrations in bronchoalveolar lavage fluid samples from Holstein calves with Mycoplasma bronchopneumonia (n = 21) and healthy controls (n = 20). The samples were obtained during bronchoscopy using a standard examination method. A total of 18 elements (aluminum, bromine, calcium, chlorine, chromium, copper, iron, potassium, magnesium, manganese, molybdenum, nickel, phosphorous, sulfur, silicon, strontium, titanium, and zinc) were detected by particle-induced X-ray emission. The average bromine, iron, potassium, magnesium, and phosphorous concentrations were higher in calves with bronchopneumonia than in controls (p < 0.05). They were found to have higher amounts of calcium and zinc, and a higher zinc–copper ratio than that in healthy calves (p < 0.001). Based on the receiver operating characteristics curves, we propose a diagnostic cutoff point for zinc–copper ratio for identification of Mycoplasma pneumonia of 8.676. Our results indicate that assessment of the elemental composition of broncholaveolar lavage fluid is a promising diagnostic tool for Mycoplasma bronchopneumonia.

Platinum concentration in a murine solid tumor treated with proton therapy combined with the platinum-based chemotherapeutic agent cisplatin [cis-diamminedichloro-platinum(II) (CDDP)] was investigated with conventional and submilli-PIXE (particle-induced X-ray emission) analyses. An enhanced therapeutic effect on tumor growth was observed in NFSa fibrosarcoma tumors in mice when proton therapy with a single dose of 30 Gy and CDDP treatment (10 mg/kg) were combined. The conventional PIXE analysis based on an internal standard method showed that the platinum concentration was 2.6 ± 0.3 µg/g for the tumor samples excised from the mice 6 h after the CDDP administration. The submilli-PIXE analysis of the tumor sections showed that spatial distribution of CDDP may be more or less uniform in the tumor. Copyright © 2011 John Wiley & Sons, Ltd.

Spatial distributions of principal elements in a NFSa fibrosarcoma tumor treated with a vascular disruption agent, AVE8062 (a derivative of combretastatin A-4) at a single dose of 40 mg/kg were evaluated on the basis of particle-induced X-ray emission (PIXE) analysis using a submillimeter-sized proton beam. We found not only potassium and sulfur concentrations at the periphery of the treated tumor but also a ring-shaped calcium distribution in the area between the inner necrotic region and the periphery. These observations were supported by results for quantitative evaluation of elemental concentrations in the tumor samples obtained from conventional PIXE analysis based on the internal standard method. It is suggested that the potassium and sulfur concentrations are related to residual viable cells at the tumor periphery which are commonly observed in VDA treatments whereas the ring of calcium concentration just inside the tumor periphery is possibly derived from hypoxia-induced response.

Concentrations of the chemotherapeutic agent, cis-diamminedichloro-platinum(II) (CDDP) in NFSa fibrosarcoma tumors were investigated using a conventional PIXE analysis on the basis of an internal standard method to study enhancement of antitumor effects caused by proton therapy combined with CDDP treatment. Results of the PIXE analysis showed that platinum concentration of the tumors treated with CDDP at a single dose of 10 mg/kg was 2.0±0.1 µg/g and persisted at the level at least 6 hours after the administration. The present study demonstrated that the presence of CDDP in the tumor caused an enhanced therapeutic effect on tumor growth delay when CDDP treatment was combined with post proton-irradiation in comparison with CDDP treatment alone or proton therapy alone.

The aim of this study was to investigate the relationships between the bronchopneumonia and mean concentrations of those trace elements in bronchoalveolar lavage fluid (BALF). Twenty-nine dogs were included this study (17 healthy dogs and 12 dogs with respiratory disease). Each BALF sample had been obtained during bronchoscope examination by use of a standardized method. The concentrations of Al, Br, Ca, Cu, Fe, K, Ni, P, Si, Sr and Zn in BALF were measured by the particle-induced X-ray emission method. We found no relationship between the bronchopneumonia and the levels of elements in the BALF, except Ca, P and Zn. The dogs with respiratory disease were found to have a large amount of Ca and Zn, and a high Ca/P and Zn/Cu ratios in BALF compared to those without respiratory disease.

Serum trace elements were analyzed by PIXE method in a patient with acute aortic dissection during the supportive process by a Nutrition support team (NST). The patient was a 53-year old male who suffered from acute aortic dissection on March 1, 2007. He received emergent Bentall operation and bow pars total displacement. Since he suffered from diarrhea as a result of hypoalbuminemia and bowel dysfunction, enteral nutrition was not sufficient to maintain adequate intake. Therefore, we also used parenteral nutrition until the 97th disease day. Afterwards, the patient managed only with enteral nutrition. We measured serum trace elements during these periods. Serum trace elements such as Fe , Zn and Se were preserved within normal ranges. On the other hand, Cu gradually decreased until the 54th disease day (44 μg/l) and stayed in the lower levels around the detection limit until the 109th disease day. Thereafter, Cu slightly increased and reached 117 μg/l on the 131st disease day (reference range of serum Cu in adult males: 700~1,300 μ g/l ). It was suggested that the serum levels of these trace elements might reflect the nutritional stages of the patient and could be useful as clinical markers for nutritional therapies.

The aim of this study was to investigate the relationships between the mean concentrations of trace elements and the severity of the second-degree atrioventricular (AV) block in the mane hair of horses. Electrocardiographs of horses were continually recorded for 6 h using a holter cardiac monitor to determine dropped ventricular beats (DVBs) which can be used as an indicator of the severity of the AV block. Mane hair Ca, Cu, Mg, and Zn concentrations were measured by the particle-induced X-ray emission method. The Zn/Cu ratio and Ca concentration in mane hair were significantly and positively correlated with the hourly DVBs in horse with a second-degree AV block (p0.01, r2=0.485; p0.05, r2=0.351, respectively). Proposed diagnostic cutoff points for hair Ca concentration and Zn/Cu ratio based on receiver operating characteristics curves analysis in detecting second-degree AV block were set at 1536 microg/g and 26.0, respectively. Those results with horse hair suggest that the evaluation of the Ca, Cu, Mg, and Zn status in mane hair by this method is strongly related to the severity of second-degree AV block and might predict the susceptibility of an individual much before the development of the symptom.

In 30 healthy persons (control), 17 renal cell carcinoma (RCC) patients, 30 hemodialysis patients (HD) and 15 healthy persons in the Takko town (Aomori prefecture), the selenium concentration in serum was determined by PIXE. The mean serum selenium concentration in healthy controls was 1.4 ± 0.6 μ mol/l , in RCC patients 1.1 ± 0.4 μ mol/l , in HD patients 1.3 ± 0.9 μ mol/l and in healthy persons in the Takko town 1.7 ± 0.7 μ mol/l . There was a significant statistical difference between RCC patients and healthy persons in the Takko town ( p < 0.05). Takko town is famous for the production of garlic. Epidemiological studies have established an inverse relationship between garlic consumption and incidence of gastric, colon, and laryngeal cancers. Garlic is a rich source of selenium in the food. Contents of selenium in normal and carcinoma fractions in kidneys of RCC patients were determined by PIXE. The results indicate that RCC patients caused significant decrease of selenium contents in the carcinoma fraction ( p < 0.05).

In hemodialysis (HD) patients, an accumulation of trace elements such as aluminum, copper, silicon and vanadium has been reported. Aluminum-caused encephalopathy and aluminum-related bone diseases are important trace element-related complications. Using particle induced X-ray emission (PIXE) we determined concentrations of aluminum, silicon, copper, zinc, selenium and bromine in sera of 29 patients with HD, 14 nondialysis patients with renal disease (RD) and 27 normal controls. The concentration of serum silicon of the patients with HD was 107.4 ± 61.3 μmol/l, which is markedly higher than that of normal controls (48.3 ± 25.8 μmol/l, p p p p

Various causal factors have been proposed for cerebral vasospasm after subarachnoid hemorrhage (SAH), such as serotonin, acetylcholine, angiotensin, thrombin and thromboxane A2. However, none of them explain the whole pathomechanism of vasospasm. To evaluate the role of trace elements on vasospasm, we have examined these sequential changes in the cerebrospinal fluid (CSF) after SAH by PIXE, and have investigated the relation between trace elements and vasospasm. We obtained the CSF samples from cisternal drainage in patients with SAH who underwent radical surgery within 48 h from the onset. The drainage was placed into basal cisterns at the end of the operation. Three sampling times (3–5, 7–9 and 12–14 days from the onset) has been scheduled because vasospasm is likely to occur from day 4 to day 14 after the onset. In this study, we focused on the levels of Mg, Ca, Mn, Al, Zn, P, Pb, Sr, Br, Co, Cu, Si, Ti, Mn,Co, Cu, Zn, Br, Sr, Mo and Pb, and we found a significantly lower level of Mg in the CSF of patients with vasospasm on days 7–9 after the onset. These results suggest that Mg in the CSF may ameliorate vasoconstriction due to Ca in the pathomechanism of vasospasm.

We made a comparative study of the elements in the periosteum on titanium plates and screws for internal bone fixation, normal periosteum, and oral mucosa by the PIXE method. We studied 11 patients, 4 men and 7 women, with mandibular fracture or facial deformity. The implanted time length of the materials in the body was 5 to 16 months. The analyzed samples were 11 periostea on the materials, 11 normal periostea and 4 oral mucosae. The results were as follows. Twenty-four essential and 11 contaminated elements were detected in the periostea on the materials as well as in the normal periostea and the oral mucosae. In the mean values of titanium and aluminum, there were significantly higher values in the periostea on the materials than in the normal periostea. The mean concentration values of the other elements did not differ significantly between that in the periostea on the materials and that in the normal periostea. The concentration of titanium in the periostea on the materials was not correlated with sex, age of the patients, or the implanted time length. However, there was a significantly higher titanium concentration value in the periostea on the plates than on the screws. Our results could indicate that the existence of a titanium element in the periostea on the materials was caused by its dissolution from the materials.

Cerebral vasospasm is a characteristic complication after subarachnoid hemorrhage (SAH), and the onset of vasospasm is a very important factor to decide the patient's outcome. Though various causal factors have been proposed for cerebral vasospasm after SAH, none of them explain the whole pathomechanism of vasospasm. To evaluate the role of trace elements in vasospasm, we have examined the sequential change in element concentration in the cerebrospinal fluid (CSF) after SAH by PIXE, and have investigated the relation between trace elements and vasospasm. We obtained the CSF samples from cisternal drainage of 17 patients with SAH who underwent radical surgery within 48 hours from the onset. The drainage was placed into basal cisterns at the end of the operation. Three sampling times (3-5, 7-9 and 12-14 days from the onset) were scheduled, because vasospasm is likely to occur from day 4 through day 14 after the onset. Cerebral angiograms were performed to classify vasospasm on day 1 and 7 after the onset. We measured 29 elements in the CSF and focused on Ca and Mg levels in this study, since Ca -influx into the smooth muscle cells is a principal mechanism of muscle contraction, and the competition between Ca and Mg is closely related to the muscle contraction. We found a significantly lower levels of Mg in the CSF of patients with vasospasm on days 7-9 after the onset. These results suggest that Mg in the CSF possibly ameliorate vasoconstriction due to Ca in the pathomechanism of vasospasm.

Elemental concentrations of human gallstones were determined by PIXE method. Gallstone samples analyzed in this report are classified into cholesterol gallstone (pure cholesterol stone and mixed stone) and pigment gallstone (black stone and calcium bilirubinate stone). The targets were prepared by paste spreading method previously reported, which enable us to determine also volatile elements, such as halogen atoms, Hg and so on, under acidic condition. Concentrations of 17 kinds of elements were determined and characteristic distributions are presented for each type of gallstone. Ca values are the highest among of them in all types of gallstones. Pigment gallstones contain significantly higher concentrations of 9 kinds of elements (Na, Mg, S, Cl, Ca, Mn, Fe, Cu, Zn) than cholesterol gallstones. Concentrations of S, Fe, and Zn in black stones are significantly higher than those in calcium bilirubinate stones. Almost all elements contained in the samples can be determined by PIXE method, and it is supposed that the elements or some of them affect formation of gallstones and their pigment. Some samples of the pigment gallstones include I, Hg, and Pb.

As sample preparation technique for PIXE analysis of brain tumor, “homogenizing method” and “chemical ashing method” have been studied and developed. It is concluded that chemical ashing method is quite appropriate for biological sample preparation. By taking samples of surgically removed brain tumor (meningioma) and brain tumor cell line (rat glioma and human glioblastoma), elemental constitution of brain tumor is compared with that of normal brain, and remarkably high concentration of Zn is found in brain tumor.

Using PIXE we determined concentrations of iron, nickel, copper, zinc, selenium, and rubidium in sera of 24 patients with acute myocardial infarction and 12 healthy controls. Rubidium*, selenium*, zinc**, nickel** and iron** concentrations are significantly (P

It has been previously shown that activated alveolar macrophage in idiopathic interstitial pneumonia (IIP) played an important role in the process of fibrosing, but the initial agent which activates alveolar macrophages has not been proved. The present study was designed to make quantitative analyses of trace elements in the lung tissue speciments of IIP patients and normal controls. Particle induced X-ray emission (PIXE) was used. The IIP group consisted of 11 patients and the Control group, which had no apparent history for dust inhalation and lung diseases, consisted of 9 subjects. More silicon and magnesium were found in the IIP group than in the normal controls. These data suggested that inhalation of silicon and magnesium might be one of the etiologic factors of IIP.

Alterations of heavy metal concentration in inoculated Sarcoma -180 after irradiation by 6MeV electron beam at the doses of 5Gy were evaluated in vivo in BALB/c mice, by Particle Induced X -ray Emission (PIXE). Analysis was performed for K, Cl, S, Fe, Zn and Cu. In the irradiation of 5Gy, the concentration of K and Cl have revealed the fast incremment at 1 hour after irradiattion and the fast decremment after that time. As for the S, Fe, Cu and Zn, their concentrations have revealed the fast incremment at 1 hour after irradiattion and the slow decremment from 1 hour after irradiation (except for slow increment from 1 hour to 1 day after).

Arsenic trioxide, As(2)O(3) (ATO), has been established to be an effective agent for treating acute promyelocytic leukemia, but its effect on solid tumors has not been fully explored. In the present study in a murine xenograft system, we found that ATO significantly inhibited tumor growth of the inoculated human hepatocellular carcinoma cell line HuH7 when administered either intravenously or intratumorally. Pathological examination revealed that ATO induced extensive cell death in the tumor. Some of the dead cells in intratumorally ATO-treated mice showed characteristic features of apoptosis, such as nuclear condensation and fragmentation, and were TUNEL-positive. The measurement of arsenic by using particle induced X-ray emission revealed that arsenic was accumulated more in the tumor than in brain, kidney or liver after the intravenous injection of ATO, which is consistent with the hemorrhagic cell death observed in ATO-treated tumor tissues. Thus, ATO appears to have potential for the treatment of solid tumors, as well as hematopoietic malignancies.

A consecutive series of 41 patients with unstable lumbo-sacral spine (spondylolysis, spondylolisthesis, disc lesion et al.) treated by a technique of posterolateral fusion resulted in a high percentage of good roentgenographic and clinical results.Twenty-nine patients were male, and twelve were female (15-50 years).The longest term follow-up was 3 years, and the shortest was ten months, the average being one year and six months.Solid union occured in 90 per cent for fusion of single intervertebral joint roentgenographicly, but only 62 per cent for two or more levels.However, a good clinical result was achieved in 83 per cent.

This paper reports two cases of loose bodies developing in the hip joints of the patients who visited our clinic because of limited movement and pain of the hip joint. They recieved operation in our hospital.Case 1: 25 year-old female has noticed the pain of her right hip joint while standing and stooping since 3 years ago. As the pain gradually increased and sometimes caused “locking”, she visited us. She was operated on, and was diagnosed as osteochondromatosis histopathologically. Now, 10 months after operation, she is working without trouble.Case 2: 23 year-old male complained of limited movement and pain of the right hip joint after traffic accident, which caused an injury of the right thigh and an eye ball rupture. Histopathological diagnosis showed osteochondritis dissecance. He got well two months after operation.