Background: Particulate matter (PM) which contains various transition metals, in particular iron which generates hydroxy radicals, can penetrate the distal regions of the lung causing respiratory and cardiovasucular diseases.Aims: The purpose of this study is to clarify adverse effects of fine PM on the alveolar epithelium using a primary culture of alveolar type II cells.Methods: PM was collected continuously for each month using Andersen samplers by the impact cascade method.PM was collected and separated into 5 fractions in accordance with diameter of the particles.Metal elements in fine PM were analyzed with an inductively coupled plasmaatomic emission spectrometer.Alveolar type II cells were isolated from specific pathogen-free Fisher 344 male rat using elastase inserting via the trachea.The cells were exposed to each subdivided fraction of PM at the concentration of 500 μg/ml for 6 hrs.At the end of the experiment, lactate dehydrogenase (LDH) released into the culture medium was determined and the percentage of LDH leakage was then calculated to reflect the cell injury by PM.Results: The cells were injured by fine PM of which the diameter are below 1.1 μm but not by larger one.The cell injury was attenuated by superoxide dismutase (SOD) and deferoxamine, a metal chelator.Conclusions: The present study demonstrates that fine indoor PM causes oxidative lung cell injury which may be closely associated with transition metals, in particular Fe, and have established the assay model for a study of the oxidative lung cell injury.

Background: Particulate matter (PM) which contains various transition metals, in particular iron which generates hydroxy radicals, can penetrate the distal regions of the lung causing respiratory and cardiovasucular diseases.Aims: The purpose of this study is to clarify adverse effects of fine PM on the alveolar epithelium using a primary culture of alveolar type II cells.Methods: PM was collected continuously for each month using Andersen samplers by the impact cascade method.PM was collected and separated into 5 fractions in accordance with diameter of the particles.Metal elements in fine PM were analyzed with an inductively coupled plasmaatomic emission spectrometer.Alveolar type II cells were isolated from specific pathogen-free Fisher 344 male rat using elastase inserting via the trachea.The cells were exposed to each subdivided fraction of PM at the concentration of 500 μg/ml for 6 hrs.At the end of the experiment, lactate dehydrogenase (LDH) released into the culture medium was determined and the percentage of LDH leakage was then calculated to reflect the cell injury by PM.Results: The cells were injured by fine PM of which the diameter are below 1.1 μm but not by larger one.The cell injury was attenuated by superoxide dismutase (SOD) and deferoxamine, a metal chelator.Conclusions: The present study demonstrates that fine indoor PM causes oxidative lung cell injury which may be closely associated with transition metals, in particular Fe, and have established the assay model for a study of the oxidative lung cell injury.

It is ambiguous whether the myocardial bridge (MB) over the coronary arteries and their branches is present in or absent from dextrocardia with situs inversus. Two cases of dextrocardia with situs inversus were accidentally found in adult Thai hearts received for researches from the Department of Anatomy, Faculty of Medicine, Chiang Mai University. They were found in 43- and 78-year-old Thai men.

Background: Particulate matter (PM) which contains various transition metals, in particular iron which generates hydroxy radicals, can penetrate the distal regions of the lung causing respiratory and cardiovasucular diseases.Aims: The purpose of this study is to clarify adverse effects of fine PM on the alveolar epithelium using a primary culture of alveolar type II cells.Methods: PM was collected continuously for each month using Andersen samplers by the impact cascade method.PM was collected and separated into 5 fractions in accordance with diameter of the particles.Metal elements in fine PM were analyzed with an inductively coupled plasmaatomic emission spectrometer.Alveolar type II cells were isolated from specific pathogen-free Fisher 344 male rat using elastase inserting via the trachea.The cells were exposed to each subdivided fraction of PM at the concentration of 500 μg/ml for 6 hrs.At the end of the experiment, lactate dehydrogenase (LDH) released into the culture medium was determined and the percentage of LDH leakage was then calculated to reflect the cell injury by PM.Results: The cells were injured by fine PM of which the diameter are below 1.1 μm but not by larger one.The cell injury was attenuated by superoxide dismutase (SOD) and deferoxamine, a metal chelator.Conclusions: The present study demonstrates that fine indoor PM causes oxidative lung cell injury which may be closely associated with transition metals, in particular Fe, and have established the assay model for a study of the oxidative lung cell injury.

To elucidate age-related changes of the first septal perforator branch (FSB), the authors investigated age-related changes of elements in the FSBs by direct chemical analysis. In addition, the effects of different arterial sizes and genders on element accumulation were investigated in the FSBs. Sixty-two formalin-fixed adult Thai hearts were used and the FSBs were carefully dissected in the hearts. After the arteries were incinerated with nitric acid and perchloric acid, element contents were determined by inductively coupled plasma-atomic emission spectrometry.

Yoshiyuki Tohno1*, Setsuko Tohno1, Takeshi Minami2, Pasuk Mahakkanukrauh1,3,4 and Noppadol Phasukdee1 1Department of Anatomy, Faculty of Medicine, Chiang Mai University, Thailand 2Laboratory of Environmental Biology, Department of Life Science, Faculty of Science and Technology, Kinki University, Japan 3Forensic Osteology Center, Faculty of Medicine, Chiang Mai University, Thailand 4Excellence in Osteology Research and Training Center (ORTC), Chiang Mai University, Thailand

It is ambiguous whether the myocardial bridge (MB) over the coronary arteries and their branches is present in or absent from dextrocardia with situs inversus.Two cases of dextrocardia with situs inversus were accidentally found in adult Thai hearts received for researches from the Department of Anatomy, Faculty of Medicine, Chiang Mai University.They were found in 43-and 78-year-old Thai men.One case of dextrocardia with situs inversus was found in gross anatomy practice of Chiang Mai University.It was found in a 62-year-old Thai man.Therefore, the authors investigated whether the MB over the left and right coronary arteries and their branches was present in or absent from three cases of dextrocardia with situs inversus.The MB was found over the distal left coronary artery in one case of dextrocardia with situs inversus.

To elucidate age-related changes of the Atrioventricular Nodal (AVN) artery, the authors investigated age-related changes of elements in the AVN artery by direct chemical analysis. In addition, the effects of different arterial origins, arterial sizes, and genders on element accumulation were investigated in the AVN arteries. Sixty-two formalin-fixed adult Thai hearts were dissected, and the following two types of the AVN artery were found: The first type was a single AVN artery arising from the Right Coronary Artery (RCA). The second type was a single AVN artery arising from the terminal part of the Left Circumflex Artery (LCX). For element analysis, both 55 and 7 AVN arteries arising from the RCA and the LCX, respectively, were used. After the arteries were incinerated with nitric acid and perchloric acid, element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that the Zn content decreased significantly in the AVN arteries with aging, but six element contents such as Ca, P, S, Mg, Fe, and Na did not change significantly with aging. Regarding the relationships among seven elements in the AVN arteries, extremely significant direct correlations were found both between Ca and Mg contents and between P and S contents, and a significant direct correlation was found between S and Mg contents. However, no significant correlation was found between Ca and P contents in the AVN arteries. To examine an effect of the different arterial origins on element accumulation, the AVN arteries were separated into the RCA and the LCX groups by the arterial origin and age-related changes of element contents were compared between two groups. It was found that there were no significant differences between the RCA and LCX groups in age-related changes of Ca and P contents. No gender differences and effect of arterial size were found in age-related changes of Ca and P contents in the AVN arteries. To elucidate whether calcification occurred in the AVN arteries in old age, both the mass ratios of Ca/P and Mg/Ca were estimated in the AVN arteries. The mass ratio of Ca/P increased progressively in the AVN arteries with Ca increase, being not constant. The mass ratio of Mg/Ca decreased gradually in the AVN arteries with Ca increase, but the average mass ratio of Mg/Ca was moderate, being 8.9% ± 0.9%. These results indicated that calcification scarcely occurred in the AVN arteries in old age, independently of the arterial origin, arterial size and gender.

To elucidate age-related changes of the sinoatrial (sinuatrial) nodal (SAN) artery, the authors investigated age-related changes of elements in the SAN artery by direct chemical analysis. In addition, the effects of different arterial origins, arterial sizes, and genders on element accumulation were investigated in the SAN artery. Fifty-nine formalin-fixed adult Thai hearts were dissected, and the following three types of the SAN artery were found: The first type was a single SAN artery arising from the right coronary artery (RCA). The second type was a single SAN artery arising from the proximal segment of the left circumflex artery (LCX). The third type was dual SAN artery arising from both the RCA and the LCX. For element analysis, both 41 single SAN arteries arising from the RCA and the LCX and 18 larger branches of dual SAN artery were used. After the arteries were incinerated with nitric acid and perchloric acid, element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that seven element contents such as Ca, P, S, Mg, Zn, Fe, and Na did not change significantly in the SAN arteries with aging. Regarding the relationships among seven elements in the SAN arteries, extremely significant direct correlations were found among P, S, Mg, and Fe contents with one exception. However, no significant correlations were found between Ca and either P or Mg contents in the SAN arteries. To examine an effect of the different arterial origins on element accumulation, the SAN arteries were separated into the RCA and the LCX groups by the arterial origin and age-related changes of element contents were compared between two groups. It was found that there were no significant differences between the RCA and the LCX groups in age-related changes of Ca and P contents. No gender differences were found in age-related changes of Ca and P contents in the SAN arteries. To elucidate whether calcification occurred in the SAN arteries in old age, both the mass ratios of Ca/P and Mg/Ca were estimated in the SAN arteries. The mass ratio of Ca/P increased progressively in the SAN arteries with Ca increase, being not constant. The mass ratio of Mg/Ca decreased gradually in the SAN arteries with Ca increase, but the average mass ratio of Mg/Ca was very high, being 49.4 ± 16.5%. These results indicated that calcification scarcely occurred in the SAN arteries in old age, independently of the arterial origin and gender.

To elucidate the action of the myocardial bridge (MB) on the coronary artery, the authors first prepared the hearts with the MB located in the middle one third of the left anterior descending (LAD) artery and then investigated element accumulation in the LAD artery of the hearts with the MB by direct chemical analysis. Eighty-four formalin-fixed adult Thai hearts were dissected and the MBs were found in 39 of 84 hearts with a total of 44 MBs. The 37 MBs were located in the middle one third of the LAD artery. To examine the action of the MB on element accumulation in the LAD artery, the hearts with the MB which was located in the middle one third of the LAD artery and was longer than 1.5 cm were used as Materials. The left main coronary (LMC) and LAD arteries were removed from these hearts successively and the isolated arteries were divided into eight to ten segments. After incineration of arteries with nitric acid and perchloric acid, seven element contents of Ca, P, S, Mg, Zn, Fe, and Na were determined by inductively coupled plasma-atomic emission spectrometry. To examine the endothelial changes of the LAD artery, the inner surface of segments of the LAD artery was observed by scanning electron microscopy. It was found that the extent of accumulation of Ca, P, Zn, and Na was not uniform throughout the LAD artery and was higher in the proximal part than in the distal part with regard to the LAD artery beneath the MB (the tunneled LAD artery). The extent of accumulation of Ca, P, Zn, and Na in the proximal part of the tunneled LAD artery was similar to that in the segments proximal to the MB, whereas the extent of accumulation of Ca, P, Zn, and Na in the distal part of the tunneled LAD artery was similar to that in the segments distal to the MB.

To elucidate compositional changes of the ligament with aging, the authors investigated age-related changes of elements in the ligamentum capitis femoris (LCF) of monkeys with a wide range of ages by direct chemical analysis. Used rhesus and Japanese monkeys consisted of 9 males and 22 females, ranging in age from newborn to 31 years (average age = 10.4 ± 10.9 years). After incineration with nitric acid and perchloric acid, element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that the P content decreased significantly in the LCFs of monkeys with aging, but other six element contents, Ca, S, Mg, Zn, Fe, and Na, did not change significantly with aging. Assuming that the P content indicated the active cell density and the S content indicated the protein amount, an age-related change of the mass ratio of P/S was examined in the LCFs. The mass ratio of P/S decreased significantly in the LCFs in childhood. Regarding the relationships among elements, significant direct correlations were found among the Ca, P, S, and Mg contents in the LCFs. It was suggested that the active cell density of the connective tissue cells might decrease significantly in the LCF in childhood.

Trace elements and the relationships among them were investigated by direct chemical analysis in three basal ganglia regions in very old age individuals and age- and gender-related differences were assessed. After ordinary dissections at Nara Medical University were finished, the caudate nucleus, putamen, and globus pallidus belonging to the basal ganglia were removed from the identical cerebra of the subjects who consisted of 22 men and 23 women, ranging in age from 70 to 101 years (average age = 83.3 ± 7.5 years). After incineration with nitric acid and perchloric acid, the element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that the Ca, P, and Mg contents increased significantly in the putamen with aging and the Mg content increased significantly in the globus pallidus with aging, but no elements increased significantly in the caudate nucleus with aging. Regarding the relationships among elements in the basal ganglia, extremely significant direct correlations were found among the Ca, P, and Mg contents in the putamen. These results suggested that slight calcification occurred in the putamen in very old age. With regard to seven elements of Ca, P, S, Mg, Zn, Fe, and Na, it was examined whether there were significant correlations among the caudate nucleus, putamen, and globus pallidus. It was found that there were extremely significant direct correlations among all of the three basal ganglia in the P content. Likewise, with regard to the Fe content, there were extremely or very significant direct correlations among all of the three basal ganglia. Regarding the gender difference in elements, it was found that the Ca content of the caudate nucleus was significantly higher in women than in men.

To elucidate the compositional changes of the amygdala with aging, the authors investigated age-related differences of elements in human amygdalae. In addition, the relationships between the amygdala and other brain regions were investigated from a viewpoint of elements. After ordinary dissections at Nara Medical University were finished, the amygdalae were removed from the cerebra of the subjects who consisted of 22 men and 23 women, ranging in age from 70 to 101 years. In addition, the hippocampus, dentate gyrus, mammillary body of the limbic system and the caudate nucleus, putamen, and globus pallidus of the basal ganglia were also removed from the identical cerebra. After the brain samples were incinerated with nitric acid and perchloric acid, the element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that both the Ca and Mg contents increased significantly in the amygdalae with aging, but the other five element contents (P, S, Zn, Fe, and Na) did not change significantly in the amygdalae with aging. Regarding the relationships among elements, very significant or significant direct correlations were found among the Ca, P, and Mg contents in the amygdalae. To explore the relationships between the amygdala and either other limbic system or basal ganglia, the correlations between seven elements of the amygdala and hippocampus, dentate gyrus, or mammillary body, and between those of the amygdala and caudate nucleus, putamen, or globus pallidus which derived from the identical cerebra, were analyzed with Pearson's correlation. It was found that regarding the four elements of Ca, P, Mg, and Fe, a close relationship existed between the amygdala and hippocampus, globus pallidus, or mammillary body.

To clarify the manner of accumulation of Ca and P in the coronary arteries, the authors divided the coronary arteries into many segments based on arterial ramification and investigated the element contents of the segments by direct chemical analysis. After ordinary dissection at Chiang Mai University was finished, the left coronary (LC) and the right coronary (RC) arteries were removed successively from the hearts of Thai subjects. The Thai subjects consisted of seven men and five women, ranging in age from 42 to 87 years (average age = 73.9 ± 13.5 years). The LC and the RC arteries were divided into 19 segments based on arterial ramification. After incineration with nitric acid and perchloric acid, element contents of the segments were analyzed by inductively coupled plasma–atomic emission spectrometry. In two cases, a significant content of Ca and P was contained only in the left anterior descending (LAD) artery (type I). In four cases, a significant content of Ca and P was contained in both the LAD and the RC arteries (type II). In five cases, a significant content of Ca and P was contained in all the LAD, the RC, and the circumflex (CF) arteries (type III). In the other one case, no significant content of Ca and P was contained in the coronary arteries. The manner of accumulation of Ca and P in the coronary arteries was classified into the three types, I, II, and III. Regarding the average content of elements in 12 cases, the average content of Ca was the highest in the segment of the LAD artery ramifying the first left diagonal artery and was higher in the proximal and distal adjacent segments of the LAD artery ramifying the first left diagonal artery, the proximal segment of the RC artery, and the proximal segment of the CF artery. To examine an effect of arterial ramification on accumulation of Ca and P, the differences in the Ca and P content between artery-ramifying and non-ramified proximal or distal segments of the coronary arteries were analyzed with Student’s t test. It was found that there were no significant differences in the Ca and P content between the artery-ramifying and non-ramified proximal or distal segments of the coronary arteries.

To elucidate characteristics of the three ligaments constituting the spring ligament complex from a viewpoint of elements, the authors investigated age-related changes of elements, relationships among their elements, relationships among ligaments in the elements, and gender differences in the three ligaments of the spring ligament complex, the superomedial calcaneonavicular (SMCN), inferoplantar longitudinal calcaneonavicular (ICN), and third or medioplantar oblique calcaneonavicular (TCN) ligaments. After ordinary dissection at Nara Medical University was finished, the SMCN, ICN, and TCN ligaments of the spring ligament complex were removed from the subjects. The subjects consisted of 10 men and 12 women, ranging in age from 62 to 99 years (average age = 80.5 ± 9.7 years). After incineration with nitric acid and perchloric acid, the element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that although the Ca and P content hardly changed in the SMCN ligament with aging, the Ca and P content in the ICN ligament increased to about three and five times higher in the 80s in comparison with the 60s, respectively, whereas in the TCN ligament, it increased about 40% and 90% higher in the 80s compared with the 60s, respectively. Regarding the relationships among elements, significant direct correlations were found among the contents of Ca, P, and Mg in all the three ligaments of the spring ligament complex. This finding was in agreement with the previous finding obtained with the three ligaments of the anterior cruciate ligament, posterior longitudinal ligament, and ligamentum capitis femoris. Whether there were significant correlations among the three ligaments of the spring ligament complex with regard to the Ca, P, S, Mg, Zn, and Fe contents was examined using Pearson's correlation. It was found that there were significant direct correlations between the SMCN and TCN ligaments in all the Ca, P, Mg, and Zn contents and also between the SMCN and ICN ligaments in both the Mg and Fe contents but not between the TCN and ICN ligaments in the six element contents. Regarding the gender difference in elements, a significant gender difference was found only in the Mg content of the SMCN ligament, being significantly higher in men than in women.

To elucidate compositional changes of the visual system with aging, the authors investigated age-related changes of elements in the optic chiasma, lateral geniculate body, and superior colliculus, relationships among their elements, relationships among their brain regions from a viewpoint of elements, and gender differences in their elements by direct chemical analysis. After ordinary dissection at Nara Medical University was finished, the optic chiasmas, lateral geniculate bodies, and superior colliculi were resected from identical cerebra of the subjects. The subjects consisted of 14 men and 10 women, ranging in age from 75 to 96 years (average age = 85.6 ± 5.9 years). After ashing with nitric acid and perchloric acid, element contents were determined by inductively coupled plasma-atomic emission spectrometry. As the result, the average content of P was significantly higher in the optic chiasma and superior colliculus compared with the lateral geniculate body. Regarding age-related changes of elements, no significant changes with aging were found in seven elements of the optic chiasma, lateral geniculate body, and superior colliculus in the subjects more than 75 years of age. The findings that with regard to the relationships among elements, there were extremely significant direct correlations between Ca and Zn contents and significant inverse correlations between Mg and Na contents were obtained in common in all of the optic chiasma, lateral geniculate body, and superior colliculus. It was examined whether there were significant correlations among the optic chiasma, lateral geniculate body, and superior colliculus in the seven elements and the following results were obtained: There were significant direct correlations between the optic chiasma and lateral geniculate body in both the P and Mg contents; there was a significant direct correlation between the optic chiasma and superior colliculus in the Fe content; and a significant direct correlation was found between the lateral geniculate body and superior colliculus in the Mg content. Regarding the gender differences in elements, it was found that both the Ca and Zn contents of the lateral geniculate body were significantly higher in women than in men.

To elucidate the relationships among the brain regions belonging to the limbic system, the authors investigated the relationships among the hippocampus, dentate gyrus, mammillary body, and fornix, using the anterior commissure as a control, from a viewpoint of elements. After ordinary dissections at Nara Medical University were finished, the hippocampi, dentate gyri, mammillary bodies, fornices, and anterior commissures were resected from identical cerebra of the subjects. The subjects consisted of 23 men and 23 women, ranging in age from 70 to 101 years (average age = 83.5 ± 7.5 years). After ashing with nitric acid and perchloric acid, element contents were determined by inductively coupled plasma-atomic emission spectrometry. With regard to seven elements of Ca, P, S, Mg, Zn, Fe, and Na, it was examined whether there were significant correlations among the hippocampus, dentate gyrus, mammillary body, fornix, and anterior commissure. It was found that there were extremely or very significant direct correlations among all of the five brain regions of the hippocampus, dentate gyrus, mammillary body, fornix, and anterior commissure in the P content. Likewise, with regard to the Fe content, there were significant direct correlations among the four brain regions belonging to the limbic system, except for the anterior commissure. In both the Ca and Zn contents, there were extremely or very significant direct correlations among the hippocampus, dentate gyrus, and mammillary body of the gray matter.

To examine whether there were gender differences in the sino-atrial node (SAN), the authors investigated the gender difference in the SAN using monkey hearts by direct chemical analysis from a viewpoint of element contents. The used rhesus and Japanese monkeys consisted of 30 males (average age=6.5±7.5 years) and 30 females (average age=12.2±10.3 years), ranging in age from newborn to 30 years. The SAN tissues were removed from the anatomical position of monkey hearts and were confirmed by means of histological observation. After ashing with nitric acid and with perchloric acid, element contents of the SANs, such as Ca, P, S, Mg, Zn, Fe, and Na, were determined by inductively coupled plasma-atomic emission spectrometry. In addition, gender differences in the right atrial walls, left ventricular walls, mitral valves, and left coronary arteries of monkeys were also investigated as controls. It was found that the P content was significantly higher in females than in males in the SANs of monkeys, but the other six element contents, Ca, S, Mg, Zn, Fe, and Na, were not significantly different between males and females in the SANs of monkeys. Regarding the P content, a similar finding was also obtained in both the right atrial walls and the left ventricular walls of monkeys, but it was not obtained in the mitral valves and the left coronary arteries of monkeys. The P content of tissue is mostly determined by the nucleic acid (DNA and RNA) content and the phospholipid content of tissue. Nucleic acids in the cell nucleus and the cytosol, and phospholipids in the cell membrane are all indicators of metabolically active cells. It is reasonable to presume that the P content in the SAN indicates the active cell density, namely, the number of active cells per volume. Therefore, there is a possibility that the active cell density of the SAN is significantly higher in females than in males.

To examine whether there were gender differences in compositional changes of the coronary artery with aging, the authors investigated the gender difference in age-related changes of elements in the left coronary arteries of Thais by direct chemical analysis. After ordinary dissections by students at Chiang Mai University were finished, the left coronary arteries were resected from Thai subjects. The Thai subjects consisted of 69 men and 34 women. The ages of the male subjects ranged from 25 to 87 years (average age = 62.6 ± 11.4 years) and of the female subjects from 24 to 86 years (average age = 59.4 ± 14.6 years). After incinerating the arteries with nitric acid and perchloric acid, the element content was determined by inductively coupled plasma–atomic emission spectrometry. The Ca and P contents tended to increase in the left coronary arteries of men with age, but the increases were not statistically significant. In the left coronary arteries of women, the Ca and P contents increased significantly and progressively with aging. In addition, the Na content increased significantly in the left coronary arteries of both men and women with aging. The differences in the average contents of Ca and P by age group were observed between the left coronary arteries of men and women. With Student’s t test, significant gender differences in the average contents of Ca and P were found in both the 40s and the 70s. The Ca and P contents of the left coronary arteries in the 40s were significantly higher in men than in women. In contrast, the Ca and P contents in the 70s were significantly higher in women than in men. These results indicated that the accumulation of Ca and P in the left coronary arteries of Thais occurred at least 10 years earlier in men than in women, but a higher accumulation of Ca and P in old age occurred in the left coronary arteries of women compared with those of men. The present study revealed that there were significant gender differences in the left coronary arteries with regard to the accumulation of Ca and P with aging. It is reasonable to presume that taking clinical findings into consideration, the gender differences in the left coronary arteries may result from hormonal and/or genetic factors rather than lifestyle factors.