Preparation of ceramic microspheres for in situ radiotherapy of deep-seated cancer

Radiotherapy is one of the most effective treatments for cancers. However, external irradiation provides only small doses to deep-seated cancers, and often causes damage to healthy tissues. It has been reported that 20-30 microm diameter 17Y(2)O(3)-19Al(2)O(3)-64SiO(2) (mol%) glass microspheres are useful for the in situ irradiation of cancers. Yttrium-89 (89Y) in this glass can be neutron bombarded to form the beta-emitter 90Y (half-life=64.1h). When injected in the vicinity of the cancer, such activated glass microspheres can provide a large localized dose of beta-radiation. The Y(2)O(3) content of the glass in the microspheres is limited to only 17 mol%. Chemically durable microspheres with a higher Y(2)O(3) content need to be developed. Phosphorus-31 (31P) with 100% natural abundance can also be activated by neutron bombardment to form the beta-emitter 32P (half-life=14.3d). Chemically durable microspheres containing a high phosphorus content are expected to be more effective for cancer treatment. We prepared pure Y(2)O(3) and YPO(4) microspheres using a high-frequency induction thermal plasma melting technique, and investigated the resulting structure and chemical durability. We successfully prepared smooth, highly spherical polycrystalline Y(2)O(3) and YPO(4) microspheres with diameters in the range 20-30 microm. Both the Y(2)O(3) and YPO(4) microspheres showed high chemical durability in saline solutions buffered at pH=6 and 7. These microspheres are expected to be more effective than the conventional glass microspheres for the in situ radiotherapy of cancer.     

Production of interferon-like substance by mouse spleen cells through contact with BHK cells persistently infected with HVJ

A virus inhibitor, an interferon-like substance, was found in the culture fluid of mouse spleen cells cocultivated with BHK-HVJ cells, the BHK cells persistently infected with HVJ, but not in the medium of cocultivation of mouse spleen cells and normal BHK cells. Neither the culture fluid of spleen cells alone nor that of BHK-HVJ cells alone was shown to contain the virus inhibitor. No virus inhibitory activity could be detected in the culture fluid of mouse spleen cells incubated with either the culture fluid of BHK-HVJ cells contained noninfectious HVJ particles or sonicated BHK-HVJ cell suspension. L cells or mouse liver cells, when cocultivated with BHK-HVJ cells, did not release a virus inhibitor.These findings suggest that mouse lymphoid cells have a capacity to produce a virus inhibitor when cocultivated with BHK-HVJ cells, and that nonlymphoid somatic cells may lack this capacity.Interposition of a Millipore filter between BHK-HVJ monolayer and mouse spleen cells or pretreatment of BHK-HVJ cells with anti-HVJ antiserum resulted in a blockade of virus inhibitor production. These findings suggest that the following sequence is necessary for the mouse spleen cells cocultivated with BHK-HVJ cells to produce the virus inhibitor: first, attachment of the spleen cells to BHK-HVJ cells and, second, recognition by the former of virus antigen(s) present on the surface of the latter.The BHK-HVJ cell membrane, isolated by sucrose density gradient centrifuge, was found to be an active inducer of the virus inhibitor. Moreover, some artificial membranous structures, such as HVJ-erythrocyte complex as well as HVJ spike-erythrocyte complex, exhibited a similar activity.This virus inhibitor induced in the present system appears to have all biologic attributes of interferon and its production might be initiated by membrane-membrane interaction between lymphoid cells and HVJ infected cells.     

Development of early apopotosis and changes in lymphocyte subsets in lymphoid organs of mice orally inoculated with nivalenol

Development of early apoptosis and changes in lymphocyte subsets were examined in lymphoid organs of female BALB/c mice after oral administration of 15 mg/kg b.w. of nivalenol (NIV), the major type B trichothecene mycotoxin, by FACS analysis. Judging from the results of viable cell count and apoptotic cell index, NIV attacked Peyer's patches first and thymus most severely. In thymus, selective damage in CD4(+)CD8(+) cells was observed at 12 and 24 h after inoculation (HAI), following the peak of apoptosis at 9 HAI. CD4(+) cells were clearly suppressed at 3 HAI in Peyer's patches, at and after 9 HAI in mesenteric lymph nodes, and 3 to 12 HAI in spleen, respectively. CD8(+) cells were also suppressed at 24 HAI in mesenteric lymph nodes and at 12 HAI in spleen, respectively. As to changes in B cell subsets, IgG(+) cells significantly decreased from 3 to 12 HAI and all B cell subsets at 24 HAI in mesenteric lymph nodes. In spleen, IgM(+) cells were suppressed at 9 HAI. On the other hand, in Peyer's patches, following clear decrease in the numbers of pan-T and pan-B cells and viable cells at 3 HAI, all B cell subsets, especially IgA(+) cells, showed a significant increase in their numbers at 9 HAI, and the numbers of IgA(+) and IgM(+) cells remained higher values than controls thereafter. Taken together, in the course of recovery from NIV-induced prominent damage in Peyer's patches at 3 HAI, interaction of NIV with Peyer's patches might result in in vivo stimulation of interleukin production at this site and result in increased proliferation and differentiation of IgA-secreting B cells at and after 9 HAI.     

Prevention of endotoxin shock by an antibody against leukocyte integrin {beta}2 through inhibiting production and action of TNF

Septic shock remains a serious disorder associated with highmortality. Accumulating evidence indicates that TNF is a majorand essential mediator of endotoxin shock. We report here thatadministration of an antibody against CD18 dramatically reducedendotoxin-induced shock inrabbits as revealed by preventionof severe hypotension, metabolic acidosis and a pathologicalchange suggestive of disseminated intravascular coagulationwith concomitant inhibition of elevation of plasma TNF activity.The anti-CD18 antibody also inhibited the hypotension inducedby administering recombinant TNF. Furthermore, an antibody againsta ligand for CD18 complexes, intercellular adhesion molecule-1,also prevented TNF-induced shock as well as endotoxin shockinrabbits. These observations suggest that adhesion of leukocytesto endothelium may be of primary importance in the action ofTNF as well as in the production of TNF in vivo and that theantibody against adhesion molecules could be of therapeuticbenefit in life-threatening septic shock in humans.     

Quantitative distribution of rat brain monoamine oxidase A by [14C]clorgyline autoradiography

The distribution of functionally active monoamine oxidase type A (MAO-A) was investigated by in vivo quantitative autoradiography using [¹⁴C]clorgyline in normal, conscious rat brain. [¹⁴C]clorgyline was synthesized by the methylation reaction of N-desmethylclorgyline using [¹⁴C]methyliodide. Sixty minutes after [¹⁴C]clorgyline administration (1.58 MBq/animal i.v.), the brains were removed and prepared for autoradiography by washing the brain sections with 5% trichloroacetic acid solution to remove the nonbinding free tracer. The amount of MAO-A was calculated from the regional acid-insoluble tissue radioactivity and the specific activity of the tracer. The highest amount of MAO-A (5.84 nmol/g tissue) was found in the locus coeruleus. The interpeduncular nucleus, habenular nucleus, fasciculus retroflexus, and solitary tract nucleus possessed over 1.6 nmol/g tissue of MAO-A. Among 23 regions of interest, the lowest amount of MAO-A (0.37 nmol/g tissue) was found in the globus pallidus. The findings of this study suggest that the pattern of MAO-A parallels both in neuroanatomical distribution and in density that of norepinephrine and serotonin innervation. The MAO-A concentration was, however, relatively low in the dopamine-related areas. This corresponded to the previous results obtained by histochemical analysis. In addition, among the white matter structures, a high amount of MAO-A was found specifically in the fasciculus retroflexus.     

Genetic contribution of tumor necrosis factor (TNF)-alpha gene promoter (-1031, -863 and -857) and TNF receptor 2 gene polymorphisms in endometriosis susceptibility

PROBLEM: Tumor necrosis factor (TNF)-alpha is a major cytokine involved in inflammatory and immune function. The aim of this study was to investigate whether polymorphisms at positions -1031, -863 and -857 in the TNF gene promoter region (TNFA) and TNF receptor type 2 gene (TNFR2) are responsible in part for genetic susceptibility to endometriosis. METHODS OF STUDY: TNFA and TNFR2 polymorphisms were determined in 123 patients with endometriosis and 165 fertile healthy women by the polymerase chain reaction (PCR) - preferential homoduplex formation assay and PCR-restriction fragment length polymorphism, respectively. RESULTS: The frequency of the TNFA-U01 haplotype was increased significantly in patients with endometriosis compared with controls (P = 0.045, OR = 1.45). The TNFA-U01 haplotype was strongly associated with HLA-B*0702. No difference was found in TNFR2 polymorphism between patients and controls. CONCLUSION: Our results indicated that TNFA promoter polymorphism was associated with susceptibility to endometriosis. However, this association was not independent of HLA-class I polymorphisms.     

ELISAs for free light chains of human immunoglobulins using monoclonal antibodies: comparison of their specificity with available polyclonal antibodies

For the measurement of human immunoglobulin free light chains (LCs) in clinical samples, highly specific assays for free LCs are required to discriminate them from LC portions of intact immunoglobulins (bound LCs). To develop specific enzyme-linked immunosorbent assays (ELISAs) for free LCs, two anti-free LC kappa and lambda monoclonal antibodies (MAbs) were raised by a mouse/mouse hybridoma technique. We compared the specificities of these two MAbs with those of six commercially available anti-free LC antisera, which are widely used in free LC immunoassays. Comparative titrations against free LCs and intact IgGs showed the MAbs had less cross-reactivity to intact IgGs, while the four of six antisera had high reactivity to intact IgGs. Using these MAbs, we developed LC kappa and LC lambda ELISAs with ranges from 7.8 to 500 micro g/l of free LCs and less cross-reactivity to intact IgGs (less than 0.12%). On the other hand, ELISAs with anti-free LC antisera showed low specificity and/or sensitivity. Thus, the use of these MAbs may provide reliable methods for specific measurements of free LCs in clinical samples.     

Effects of focus on working memory

Effects of focus on a target word during performance of the reading span test (RST) were investigated. A focus word in the sentence was defined as the most critical word with a core meaning to integrate the sentence. Two kinds of RST were compared. One was focus-RST (F-RST) in which the target word to be maintained was a focus word of the sentence. The other was a non-focus-RST (NF-RST) in which the target word was not a focus word of the sentence, although the sentence did contain a focus word. Results showed that RST scores were found to be higher for F-RST than for NF-RST. Moreover, the effect of focus was proved to be dominant for low span subjects. Intrusion errors also increased in NF-RST. Sentence length effect, however, was not found. The results showed that low span subjects had severe deficits in making and updating the focus, which is critical for sentence comprehension.     

Acquired gain of an X chromosome as the sole abnormality in the blast crisis of chronic neutrophilic leukemia

Chronic neutrophilic leukemia (CNL) is a rare myeloproliferative disorder characterized by sustained neutrophilic leukocytosis and absence of the Philadelphia chromosome. Most patients with CNL have normal karyotypes, and no specific cytogenetic abnormality has been identified. We report here a patient with CNL that evolved to myeloid blast crisis. A 73-year-old man was admitted to the hospital because of marked leukocytosis (leukocyte count 112.5 x 10(9)/L with 91% segmented neutrophils) and massive hepatosplenomegaly that was diagnosed as CNL with a normal karyotype. After treatment with hydroxyurea for 7 months, the disease progressed to a blast crisis. Bone marrow showed myeloid hyperplasia with 21% myeloblasts, 15% promyelocytes, and marked dysplastic changes of neutrophils. Blastic cells were positive for CD10, CD13, CD14, CD33, CD34, and HLA-DR. Chromosome analysis of the bone marrow cells showed 46,XY,+X in all 20 metaphase spreads. We reviewed 15 cases of CNL terminating in the blast crisis and confirmed that all cases transformed into myeloid crises and had poor prognoses. Furthermore, to our knowledge, this is the first case showing the acquired gain of an extra X chromosome as a sole abnormality in CNL. The gain of an extra X chromosome may play an important role in the progression from chronic phase to the blast crisis of CNL.