Development of a voxel phantom of Japanese adult male in upright posture.

A Japanese voxel phantom in upright posture, JM2, has been developed on the basis of CT images of a healthy Japanese adult male. Body characteristics of JM2 were compared with those of the supine voxel phantom, JM, previously developed using CT images of the same person. Differences were found in the shapes of the spine and lower abdomen and the locations of several organs such as kidneys, liver and stomach between the two phantoms. Specific absorbed fractions (SAFs) for 24 target and 11 sources organs were calculated for monoenergetic photon ranging from 0.01 to 4 MeV. It was found that the SAFs for the kidneys as source organ and the lower large intestine wall as target organ in JM2 were significantly higher than those in JM for all photon energies. The differences of the SAFs between the two phantoms were attributed to the differences in the organ distance and organ geometry depending on the posture.

Japanese adult male voxel phantom constructed on the basis of CT images.

A Japanese adult male voxel (volume pixel) phantom (hereinafter referred to as the JM phantom) was constructed on the basis of CT images of a healthy Japanese adult male volunteer. Body characteristics of the JM phantom were compared with those of a voxelised MIRD5 type phantom and a Japanese adult male voxel phantom which was previously developed. The voxel size of the JM phantom is 0.98 x 0.98 x 1 mm(3). The shapes of the organs of the JM phantom, even for small or complicated organs, such as thyroid and stomach, are more realistically reproduced as compared with the previous Japanese voxel phantom (voxel size: 0.98 x 0.98 x 10 mm(3)). Photon self-absorbed fractions (self-AFs) for brain, kidneys, spleen, pancreas, thyroid and urinary bladder wall of JM were evaluated and were compared with those of the other phantoms. In consequence, it was suggested that the mass, shape and thickness of organs are important factors for the determination of self-AFs.

A critical role of T-cell receptor gamma/delta cells in antibacterial protection in mice early in life.

Although it is generally assumed that T-cell receptor (TCR) gamma/delta cells participate in protection against intracellular microbial pathogens, their impact remains controversial. In our study, young (14-day-old) mice lacking TCRgamma/delta cells were far more susceptible to Listeria monocytogenes than wild-type (WT) mice of the same age. The number of interferon gamma (IFN-gamma) producers responsible for antilisterial resistance was significantly higher among natural killer (NK)1(+) TCRgamma/delta cells than among NK1(-) TCRgamma/delta cells. Endogenous IFN-gamma neutralization increased susceptibility of young WT mice to L. monocytogenes infection. Liver was a major residence of peripheral NK1(+) TCRgamma/delta cells, whereas NK1(-) TCR gamma/delta cells were broadly distributed in various lymphoid organs. Numbers of both NK1(+) and NK1(-) TCRgamma/delta cells increased in the liver of WT mice prior to TCRalpha/beta cells and represented a substantial population in early life (14 days after birth). Virtually all NK1(+) TCRgamma/delta cells expressed activation markers, whereas substantial numbers of NK1(-) TCRgamma/delta cells showed a naive phenotype. We conclude that TCRgamma/delta cells play a critical role in protection against L. monocytogenes in the early life of mice, probably because their TCRalpha/beta cell compartment is not fully competent. For this antibacterial function, we assign NK1(+) TCRgamma/delta cells a more important role than their NK1(-) cognates.

Small molecular mass inhibitor of growth of MDCK cells derived from pig spinal cord.

We have discovered cell growth inhibitory activity in a salt extract of pig spinal cords. The growth inhibitory factor was purified by gel-filtration, ion-exchange and high performance liquid chromatography. Incubation of MDCK cells with the inhibitor arrested their locomotion within half an hour, suppressed their proliferation, and caused them to become round. The round cells that were still attached to the culture plate were alive. Upon removal of the inhibitor these cells flattened out and resumed locomotion and proliferation. The inhibitor was 100 times less effective on CHO-K1 cells. The reversible effects of the inhibitor on MDCK cells and its little effects on CHO-K1 cells indicate that the inhibitory activity is not due to a non-specific toxic mechanism. The inhibitor was both heat-stable and resistant to several chemical treatments, including proteases. Its behavior upon ion exchange chromatography suggested that it was positively charged at neutral pH, whilst its molecular mass was estimated to be 350 or larger by gel-filtration FPLC analysis. The inhibitory fraction reacted extensively with fluorescamine, suggesting that the inhibitory factor has amine groups, which are a possible candidate for its positive charges. Since spermine and spermidine, unlike the inhibitor in the present study, irreversibly inhibited the growth of the MDCK cells, the inhibitory activity in the present study is thus not due to contamination by these polyamines. Our experiments also support that the inhibitor is not a peptide.

Simple and low-cost tele-nuclear medicine conference system with the e-mail protocol.

PURPOSE: Because of the recent innovative growth in computer technology, digital imaging, and the Internet, we can take advantage of these facilities for education and clinical work in nuclear medicine. We developed a tele-nuclear medicine conference system with electronic mail (e-mail) on the Internet. METHODS: Twenty-one physicians (20 radiologists, 1 neurologist), 6 technologists and 2 medical students in six university hospitals (Japan 5, Canada 1), 5 local hospitals in Japan participated in this project. We used digital still cameras (330 k pixels) equipped with a floppy disk drive and 10 x optical zoom to digitize images with JPEG compression (640 x 480 matrix). The images were attached to e-mail messages (containing a brief description of each case). The mail was sent simultaneously to all members on the mailing list. Scintigram and SPECT images as well as other radiological images were sent by e-mail. Reply mails about each case were sent to all members via the mailing list. RESULTS: During a period of 6 months, 18 cases (tumor/infection: 7, bone: 6, cardiovascular: 1, neurology; 3, endocrine: 1) with 144 e-mails (average 5.6/case) were submitted to the conference. The average period of discussion was 15.6 days. The number of attached images was 1 to 9 (average, 4.2/e-mails). JPEG compression rate was 1/10 to 1/20. The quality of the images was good enough for discussion. Some cases required additional images for further discussion. CONCLUSION: Our tele-nuclear medicine conference with an electronic mailing list and digital camera was simple and low-cost. The conference system was useful for education and clinical work.

Clinical features of hypertrophic cardiomyopathy caused by a Lys183 deletion mutation in the cardiac troponin I gene.

BACKGROUND: Mutations that cause hypertrophic cardiomyopathy (HCM) have been identified in 9 genes that code proteins in the sarcomere. Previous reports have demonstrated that cardiac troponin I (cTnI) gene mutations may account for familial HCM; however, the clinical characteristics and prognosis of patients with HCM caused by cTnI gene mutations are not known. METHODS AND RESULTS: We analyzed cTnI gene mutations in 130 unrelated probands with HCM and their families to clarify the genotype-phenotype correlations. We identified 25 individuals in 7 families with a Lys183 deletion (Lys183 del) mutation in exon 7 of the cTnI gene. The disease penetrance in subjects aged >20 years was 88% by echocardiography and 96% by ECG. Sudden death occurred in 7 individuals of 4 families at any age. Overall, 7 (43.8%) of 16 individuals aged >40 years had left ventricular systolic dysfunction, and 3 (18.8%) displayed dilated cardiomyopathy-like features. Of affected individuals, 4 of 5 individuals aged >40 years followed by echocardiography showed septal thinning and decreased fractional shortening during >5 years of follow-up. CONCLUSIONS: The Lys183 del mutation in the cTnI gene in patients with HCM is associated with variable clinical features and outcomes. HCM caused by the Lys183 del mutation has a significant disease penetrance. This mutation is associated with sudden death at any age and dilated cardiomyopathy-like features in those aged >40 years. However, it remains unclear whether screening of families with HCM for this mutation will be useful in patient management and counseling.

Cardiac dysfunction and long-term prognosis in patients with nonobstructive hypertrophic cardiomyopathy and abnormal (123)I-15- (p-Iodophenyl)-3(R,S)-methylpentadecanoic acid myocardial scintigraphy.

To evaluate the relationship between myocardial scintigraphic abnormalities based on (123)I-radioiodinated 15-(p-iodophenyl)-3(R, S)-methylpentadecanoic acid (BMIPP) uptake and cardiac function and the relationship between these abnormalities and long-term prognosis in patients with hypertrophic cardiomyopathy (HCM), 27 patients with nonobstructive HCM underwent BMIPP myocardial scintigraphic study, echocardiography, and exercise radionuclide study. Based on the extent of BMIPP scintigraphic defects, the patients were divided into two groups: Group A (n = 19) patients had no or small defects, and group B (n = 8) patients had moderate to large defects. Cardiac events were recorded over an average period of 64 months. The left ventricular end-diastolic and end-systolic dimensions were significantly greater in group B than in group A. The fractional shortening in group B was less than in group A (p = 0.0002). The BMIPP score and fractional shortening at rest correlated significantly (p < 0.05). The BMIPP score and the change in ejection fraction between rest and peak exercise correlated significantly (p < 0.05). While only 1 cardiac event occurred in the 19 patients in group A during a mean follow-up period of 64 months, 6 cardiac events occurred in the 8 patients in group B. The 84-month event-free survival rate was 94.4% in group A and 14.6% in group B (p < 0.01). These results suggest that patients with HCM and moderate to large defects as assessed by BMIPP myocardial scintigraphy have decreased cardiac function and a poor prognosis.

Exercise-induced ST-segment depression and systolic dysfunction in patients with nonobstructive hypertrophic cardiomyopathy.

BACKGROUND: ST-segment depression is common in patients with hypertrophic cardiomyopathy (HCM). However, it is not clear whether exercise-induced ST-segment depression in patients with HCM and patent coronary arteries is associated with changes in left ventricular function. METHODS: Left ventricular function was continuously evaluated in 53 patients with nonobstructive HCM during supine ergometer exercise with a radionuclide ventricular function monitor equipped with a cadmium telluride detector. On the basis of the ST-segment changes during exercise, the patients were divided into 2 groups: group N had no ST-segment depression, and group D had >/=0.1 mV ST-segment depression. RESULTS: Exercise duration, blood pressure, heart rate, and rate-pressure product during exercise did not differ between the 2 groups. End-diastolic volume at rest and at peak exercise did not differ between groups D and N. In contrast, the end-systolic volume in group N decreased during exercise, whereas in group D it increased. As a result, the left ventricular ejection fraction in group D decreased from 70% +/- 7% to 59% +/- 15% (P <.0001), whereas ejection fraction in group N increased from 65% +/- 8% to 71% +/- 11% (P =.0002). There was a strong correlation between exercise-induced ST-segment depression and changes in ejection fraction from rest to peak exercise (P <.0001). CONCLUSIONS: These results suggest that the exercise-induced ST-segment depression seen in patients with nonobstructive HCM is associated with systolic dysfunction during exercise.

Cardiac sympathetic activity in the asymmetrically hypertrophied septum in patients with hypertension or hypertrophic cardiomyopathy.

BACKGROUND: In patients with essential hypertension (HT), proportional (symmetric) left ventricular hypertrophy (LVH) is common. In contrast, hypertrophic cardiomyopathy (HCM) is characterized by disproportional LVH and, in particular, asymmetric septal hypertrophy (ASH); however, some hypertensive patients also develop ASH. It has not been determined whether such cases represent a distinct type of hypertensive LVH or HCM combined with hypertension. HYPOTHESIS: The study was undertaken to evaluate sympathetic activity in the interventricular septum in patients with HT and ASH or in patients with HCM. METHODS: The patients were evaluated by I-123 meta-iodobenzylguanidine (MIBG) and thallium-201 (201Tl) single-photon emission computed tomography (SPECT), respectively. They were divided into three groups: patients with essential HT and symmetric septal hypertrophy (Group A), patients with HT and ASH (Group B), and patients with HCM and ASH (Group C). RESULTS: Compared with the lateral wall, early uptake of MIBG in the septum was significantly higher in Group B than in Group A, but not significantly different between Groups A and C. Compared with the lateral wall, early uptake of 201Tl in the septum did not differ among the three groups. No significant difference in the MIBG clearance in the lateral wall was seen among the three groups. By contrast, MIBG clearances in the septum and apex were significantly greater in Group C than in Groups A and B. There was an inverse correlation between systolic thickening and MIBG clearance in the septum. CONCLUSION: These findings suggest that sympathetic activity in the septum differs between patients with HT and ASH and patients with HCM.

Early and delayed Tc-99m ECD brain SPECT in SLE patients with CNS involvement.

We compared early and delayed Tc-99m ECD SPECT scans in 32 SLE patients (Group 1, definite neuropsychiatric disorders; Group 2, minor neurologic symptoms or normal) with those of normal controls by visual inspection and semi-quantitative evaluation. With visual interpretation, 13 out of 14 patients in Group 1 (93%) and 7 out of 18 patients in Group 2 (39%) had diffuse uneven decrease in early scans. Seven patients in Group 2 (39%) who had normal early scans demonstrated focal decrease in the medial frontal lobe in delayed scans. With cerebral region to cerebellar ratios, in early scans, the medial frontal lobe in Group 1 and Group 2 was significantly lower than in normal controls, and lateral frontal lobe and occipital lobes in Group 1 were significantly lower than in normal controls. Nevertheless, in delayed scans, every cortical region except for the parietal lobe in Groups 1 and 2 was significantly lower than in normal controls. The retention rates in all regions in SLE patients were significantly lower than in normal controls. No case showed SPECT improvement on follow-up studies in either group in spite of clinical improvement. Delayed Tc-99m ECD brain SPECT of high sensitivity might be useful in detecting CNS involvement. Although the SPECT findings did not correlate with the neuropsychiatric symptoms, early and delayed Tc-99m ECD SPECT seems to provide useful objective diagnostic information in SLE patients.

A protein friction model of the actin sliding movement generated by myosin in mixtures of MgATP and MgGTP in vitro.

The sliding movement of an actin filament generated by myosin heads with MgGTP bound is much slower than that by those with MgATP bound. Nonetheless, there is a report that the actin sliding velocity at low (11-21 microM) MgATP concentrations is increased by the addition of MgGTP in a range of 1-3 mM, although the actin sliding velocity at these MgATP concentrations is larger than the maximum sliding velocity attained in the presence of MgGTP alone. The convex rise in the velocity was called "mutual sensitization of MgATP and MgGTP" in the report. Here we propose a theoretical model to account for the mutual sensitization of MgATP and MgGTP. The model is an extension of a protein friction model, accommodating the presence of two different substrates and assuming the presence of motile and non-motile myosins. This new model is in accord with the characteristics of the actin/myosin sliding movement experimentally observed in mixtures of MgATP and MgGTP. Comparison of the model with the experimental results implies that the non-motile and motile myosins are those with the "converse and correct" orientations of their heads with respect to the direction of the actin sliding movement in vitro.

Induction of IFN-gamma-producing CD4+ natural killer T cells by Mycobacterium bovis bacillus Calmette Guérin.

The CD4+ natural killer (NK)T cells in the liver are potent IL-4 producers and hence may promote Th2 cell development. Following Mycobacterium bovis bacillus Calmette Guérin (BCG) infection, IL-4-producing CD4+ NKT cells become undetectable in liver mononuclear cells of normal density (interface between 40 and 70% Percoll) by flow cytometry. The present study shows that M. bovis BCG infection changes the density of liver CD4+ NKT cells and shifts cytokine production from IL-4 to IFN-gamma. The number of CD4+ NK1+ TCR alpha/beta(intermediate) cells increased in the low-density fraction (<40% Percoll density gradient) in parallel to the reduction of this cell population in the fraction of normal density. The number of IL-4-producing cells, however, was small and high frequencies of IFN-gamma-secreting cells were identified in the low-density fraction after TCR/CD3 ligation. Accordingly, selected low-density CD4+ NKT cells encompassed high numbers of IFN-gamma producers and minute numbers of IL-4-secreting cells. Induction of low-density CD4+ NKT cells by M. bovis BCG was abrogated by endogenous IL-12 neutralization which also caused increased bacterial growth in the liver. We assume that M. bovis BCG infection changes cytokine secretion by the CD4+ NKT cell population from IL-4 to IFN-gamma through IL-12 induction. Thus, CD4+ NKT cells may contribute to host resistance against intracellular bacteria prior to conventional IFN-gamma-producing Th1 cells.

Transient control of interleukin-4-producing natural killer T cells in the livers of Listeria monocytogenes-infected mice by interleukin-12.

Unconstrained development of gamma interferon (IFN-gamma)-secreting natural killer (NK) cells and T helper (Th) 1 cells is central to protection against Listeria monocytogenes. In contrast, interleukin 4 (IL-4) is considered harmful. IL-12 produced by infected macrophages promotes, and IL-4 interferes with, protective antilisterial immunity. The liver NK T lymphocytes, which are a potent source of IL-4, are downregulated at an intermediate stage of listeriosis. Here we demonstrate that endogenous IL-12 participates in the control of IL-4-producing liver NK T lymphocytes during listeriosis. The effects of L. monocytogenes infection on IL-4-producing liver NK T lymphocytes were reversed by antibody neutralization of IL-12 but not of IFN-gamma or tumor necrosis factor alpha (TNF-alpha). IL-4 production by liver NK T lymphocytes was virtually unaffected by heat-killed L. monocytogenes (HKL). Viable L. monocytogenes markedly increased the numbers of IL-12 producers in livers in parallel with an increase in macrophage numbers, whereas HKL failed to do so with similar efficiency. These results indicate that in the liver endogenous IL-12 improves protective immunity against listeriosis by downregulating IL-4-producing NK T lymphocytes. Moreover, our findings that HKL have a low level of IL-12-inducing activity and fail to control IL-4-producing NK T lymphocytes in the liver are consistent with the lesser protective capacity of HKL compared to that of live listeriae.

TCR-mediated target cell lysis by CD4+NK1+ liver T lymphocytes.

In the liver, an unusual T lymphocyte population exists with the intriguing phenotype CD4+NK1+ TCR alpha beta int. Thus far, functions of these lymphocytes remained elusive. Recently, however, CD4+NK1+ liver T lymphocytes have been shown to produce cytokines. Here we show that sorted CD4+NK1+ liver lymphocytes from naive mice lyse target cells after TCR alpha beta or CD3, but not TCR gamma delta, engagement. Liver lymphocytes from beta 2-microglobulin-deficient gene disruption mutant mice failed to express such cytolytic activities and in vivo treatment with anti-NK1.1 mAb or anti-CD4 mAb, but not anti-CD8 mAb, markedly reduced target cell lysis. In vivo administration or rIL-12 impaired TCR alpha beta-mediated target cell lysis by liver lymphocytes. A similar down-regulation of cytolytic activities was observed with liver lymphocytes from mice infected with Listeria monocytogenes or Mycobacterium bovis BCG, which are potent IL-12 inducers. We anticipate (i) that cytolytic CD4+NK1+ T lymphocytes contribute to immunosurveillance of inflammatory processes in the liver and (ii) that they are influenced by IL-12.

CD8 alphabeta+ TCR alphabeta(intermediate) lymphocytes expressing skewed TCRVbeta repertoire in the liver of aged athymic nu/nu mice.

Principally, TCRalphabeta cells differentiate and mature in the thymus. However, evidence has accumulated to suggest that some TCRalphabeta cells develop extrathymically. Such cells have been identified in athymic nu/nu mice that are devoid of a functional thymus. They appear relatively late in ontogeny and randomly increase in number as a function of age. Recently, the liver has been suggested as a candidate site for thymus-independent development of T cells. Here we show that CD8alphabeta+ T cells that express TCRalphabeta at intermediate intensity (TCRalpha(beta)int) are preferentially localized in the liver of nu/nu mice. This population encompassed cells expressing lectin cell adhesion molecule-1 and/or IL-2Rbeta and cells lacking either of these surface molecules. The CD8alphabeta+ TCRalpha(beta)int liver lymphocytes in nu/nu mice also differed in their LFA-1 expression marginally, whereas in C57BL/6 mice all CD8alphabeta+ TCRalpha(beta)int liver lymphocytes expressed LFA-1 at low intensity. Although the TCRVbeta repertoire markedly differed among individual animals, the CD8alphabeta+ TCRalpha(beta)int liver lymphocytes in nu/nu mice preferentially used TCRVbeta5 and/or TCRVbeta8. These findings show that CD8alphabeta+ TCRalpha(beta)int cells develop in the liver of nu/nu mice and that they arise randomly.

Bacille Calmette Guérin and interleukin-12 down-modulate interleukin-4-producing CD4+ NK1+ T lymphocytes.

Early production of interleukin-12 (IL-12) by macrophages and of IL-4 from CD4+ NK1+ T cells influence development of the acquired immune response against infectious agents, namely differentiation of interferon-gamma-secreting T helper 1 (Th1) cells against intracellular pathogens and of IL-4-producing Th2 cells against helminths. Evidence has been presented for transient convertibility of Th1 and Th2 cells in the presence of the polarizing cytokines IL-4 or IL-12, respectively. Moreover, it is likely that IL-4 dominates over IL-12, suggesting that Th2 cell development is preferred in the presence of both cytokines. Mycobacterium bovis Bacille Calmette Guerin (BCG) and IL-12 are potent inducers of Th1 responses. Here we show that BCG and IL-12 down-modulate IL-4-producing CD4+ NK1+ TCR alpha/beta(intermediate) liver lymphocytes. Our data provide further insights into the mechanisms by which BCG and IL-12 may promote unrestricted development of Th1 responses in vivo: BCG and IL-12 not only provide the positive stimuli for Th1 cell differentiation, but also interfere with antagonizing signals.

Proteolytic activation of protein kinase C delta by an ICE/CED 3-like protease induces characteristics of apoptosis.

Recent studies have shown that protein kinase C (PKC) delta is proteolytically activated at the onset of apoptosis induced by DNA-damaging agents, tumor necrosis factor, and anti-Fas antibody. However, the relationship of PKC delta cleavage to induction of apoptosis is unknown. The present studies demonstrate that full-length PKC delta is cleaved at DMQD330N to a catalytically active fragment by the cysteine protease CPP32. The results also demonstrate that overexpression of the catalytic kinase fragment in cells is associated with chromatin condensation, nuclear fragmentation, induction of sub-G1 phase DNA and lethality. By contrast, overexpression of full-length PKC delta or a kinase inactive PKC delta fragment had no detectable effect. The findings suggest that proteolytic activation of PKC delta by a CPP32-like protease contributes to phenotypic changes associated with apoptosis.

Force production by chemically crosslinked myosin-actin crossbridges in rabbit skinned fibers in response to MgATP depletion.

In order to study the contractile property of myosin crossbridges attached to thin filaments, myosin heads were crosslinked to the filaments at their interface in single skinned rabbit psoas fibers with a zero-length chemical crosslinker, 1-(3-dimethylamino-propyl)-3-ethylcarbodiimide (EDC). The results obtained show that a partially crosslinked single fiber produces a large rigor-like force when MgATP is depleted from the myofibrillar space. Such crosslinked fibers contain two types of crosslinked myosin heads: one with one of the two heads of the myosin molecule crosslinked to actin with the other head uncrosslinked; the other has both heads crosslinked to actin. The results of this work suggest that a crosslinked myosin head of the former type produces a much larger force than the latter type.