Relationships between suppressor macrophages and macrophage precursors in the spleens from tumor-bearing mice

Suppressor macrophages (Mφ) which can inhibit mitogen-induced lymphocyte proliferation appeared in the spleens of mice bearing transplanted MC-A fibrosarcoma cells. An analysis of the ontogeny of such Mφ revealed additional suppressor activity directed against macrophage stem cells. Treatment of spleen cell suspensions with carbonyl iron followed by centrifugation removed suppressor Mφ but did not deplete Mφ-colony forming cells (M-CFC) which could be demonstrated in soft agar culture in L-cell conditioned medium (LCM). Untreated spleen cells had normal numbers of M-CFC; phagocyte-depleted mononuclear cells showed a threefold increase in M-CFC 14 days after subcutaneous inoculation of 10⁶ MC-A cells per mouse. Further increases in M-CFC were also evident in similar preparations on Days 21 and 28 when the M-CFC concentration reached a maximum of eight times the normal level. The Mφ which developed from the M-CFC grown in the presence of LCM were later shown to have indomethacin-sensitive suppressor activity suggesting the mediation of this phenomenon by prostaglandins. These observations suggest that locally produced phagocytic suppressor Mφ from the spleens of tumorbearing mice play important roles not only as inhibitors of lymphocyte proliferation as reported earlier, but also as regulators of monocyte-Mφ production.     

The establishment of a human myeloma cell line elaborating lambda-light chain protein

A human myeloma cell line (KMM-56) producing lambda-light chain protein was established in vitro by cultivation of the cells in the pleural effusion obtained from a patient with IgD-lambda-myeloma. The cells proliferate in suspension and do not aggregate or attach to the culture dish. Surface marker analysis revealed that the cells were negative for E-rosette, and surface immunoglobulin. Immunoelectrophoresis, immunodiffusion, and immunofluorescence with various antibodies demonstrated no heavy chains, while lambda-light chains were detected in the cytoplasm of the cells. Using the immunodiffusion technique, only lambda-light chains were detected in the frozen and thawed cell extract, the concentrated supernatant of the cell culture, and the urine of the patient. Electron microscopic examination revealed the plasmablastoid appearance of the cells. This cell line may be useful for future studies of human immunoglobulin genes and for the material of human-human hybridoma, which could produce monoclonal human immunoglobulin.     

Characterization of the early and late stages of myelomonocytic leukemia induced by Friend helper-independent virus F-MuLV: isolation and induction of Friend myelomonocytic leukemic cell lines

With the use of cloned helper-independent Friend leukemia virus (F-MuLV), we have induced a high incidence (approximately 70%) of myelomonocytic leukemia in mice resistant (Fv-6rr or Fv-6rs) to erythroleukemia induction by this virus. The spleen cells from these mice (DBA/2 or BALB/c X DBA/2) were found to contain a high level of progenitor cells capable of forming granulocytic and macrophage colonies (CFU-GM). These CFU-GM, however, were different from those in the spleens of uninfected mice, as they were either very sensitive to or independent of conditioned medium. No erythroid progenitor bursts (BFU-E) or precursor (CFU-E) cells were detected in the spleens of these diseased animals. If these mice with myelomonocytic leukemia were kept alive by transfusion of red blood cells from uninfected mice, tumorigenic cell lines, capable of being transplanted, into adult mice can be isolated. Three such cell lines TTA-1, TTA-3, and TTA-9 have been established, and they retain their morphology of monocytes and macrophages as well as being positive for the monocyte-specific stain alpha-naphthyl-acetate esterase. These myelomonocytic leukemia cell lines can also be induced in culture by spleen cell-conditioned medium to differentiate into macrophages. Other conditioned media such as L-cell-conditioned medium, phytohemagglutinin-stimulated leukocyte-conditioned medium, and WEHI-3 conditioned medium were less effective in their abilities to stimulate differentiation in these myelomonocytic leukemia cell lines.     

Snow cover manipulation in agricultural fields: as an option for mitigating greenhouse gas emissions

Short-term N₂O emission occurs in relation to snowmelt within seasonally frozen soil. To understand the effects of changing winter climates on the N₂O flux, snow cover manipulation experiments are useful. In Japan, snow cover manipulation is practiced by farmers to improve agricultural yield and is executed either by applying a broadcast of blackish agent onto the snow cover, which leads to faster snow-melting thereby extending the crop-growing season, or by snow cover removal/re-accumulation, leading to an enhanced soil frost depth for weed management. Implementation of these practices involves using an amount of fossil fuel, in addition to influencing soil-derived N₂O emissions, therefore, the load factors of snow cover management practices per unit area of agricultural field were estimated in this study. Field data including micrometeorological conditions, ground surface flux of N₂O, and amount of fossil fuel consumed during machinery operation for management practices, were obtained at two sites in Hokkaido over 2 years (2008–2010). Fuel consumption for the field spreading was found to be unexpectedly small (0.017 Mg CO₂ eq ha^?1). It was therefore suggested that acceleration of snowmelt may have the potential to reduce net greenhouse gas emissions if the agent used is a low-degradable C-rich material, such as charcoal. For soil frost control, the fossil fuel consumption during a set of snow cover removal/re-accumulation (estimated as 0.052 Mg CO₂ eq ha^?1) is discussed, together with the relationship between possible mechanisms causing stimulation of N₂O production in frozen soil and inherent large differences in N₂O flux among sites.     

Olfactory receptor cell responses of pigeon to some odors

A preparation has been developed in the pigeon which allows recording of the electrical activity from an olfactory nerve twig containing the nonmyelinated axons of a small group of olfactory receptor cells. The pigeon's response to n-amylacetate is vigorous and stable, like that of other air-breathing animals. Responses in the olfactory receptor cells in the pigeon increased in magnitude with increase in the odor concentration. An olfactory nerve twig produced a different magnitude of responses to the various odor stimuli. When an odor stimulation was applied to the olfactory mucosa, the two different olfactory nerve twigs which were separated from the same olfactory nerve bundle produced a different magnitude of responses. The differences may be dependent on several factors.     

Effects of chest wall vibration on breathlessness during hypercapnic ventilatory response

Vibratory stimulation applied to the chest wallduring inspiration reduces the intensity of breathlessness, whereas thesame stimulation during expiration has no effect or may increasebreathlessness. The purpose of the present study was to determinewhether vibration reduced the intensity of breathlessness duringprogressive hypercapnia with and without the addition of an externalresistive load. A second objective was to see whether the mouthocclusion pressure at 0.2 s(P_0.2) was reduced by thevibratory stimulation. Hypercapnic ventilatory response was conductedin 10 healthy male volunteers with simultaneous measurement of visualanalog scale, P_0.2, and minuteventilation. Hypercapnic ventilatory response was performed andrandomly combined with or without vibratory stimulation (100 Hz) aswell as with or without inspiratory load. With inspiratory load,in-phase vibration did not cause any significant changes in the slopesof P_0.2 and minute ventilation toCO₂, whereas the slope of visualanalog scale to CO₂ significantlydecreased from 0.47 ± 0.15 to 0.34 ± 0.11 (SE) cm/Torr(P < 0.05). We conclude thatin-phase vibration could decrease the slope of breathlessness elicitedby inspiratory load combined with hypercapnia without changing motoroutput.

     

Preparation of a stable subunit of Japanese elderberry (Sambucus sieboldiana) bark lectin and its application for the study of cell surface carbohydrates by flow cytometry.

A stable subunit of Sambucus sieboldiana bark lectin (MSSA) was prepared by selective reduction of disulfide bridges between the subunits and alkylation with 4-vinylpyridine. Amino acid analysis of MSSA revealed that 1.4 cysteine residues per subunit were selectively modified. MSSA failed to agglutinate rabbit erythrocytes and precipitate fetuin. However, MSSA retained the ability to bind to fetuin, as detected by ELISA. Neu5Ac alpha 2-6lactose inhibited the binding to fetuin of both SSA and MSSA. Flow cytometric analysis showed that human histocytic lymphoma U937 cells were clearly stained with FITC-labeled MSSA (FITC-MSSA) without any detectable agglutination and that this staining was almost completely inhibited by the addition of Neu5Ac alpha 2-6lactose (2 mM). Treatment of U937 cells with native FITC-SSA at the sub-agglutinating concentration (0.3 microgram/ml) showed much poorer fluorescence intensity than that of MSSA, suggesting that MSSA is an invaluable tool for the detection of cell surface glycoconjugates containing NeuAc alpha 2-6Gal/GalNAc sequences by flow cytometry.     

Epigastrius with omphalocele--report of a case

An epigastrius (parasitic twinning in the epigastrium) was delivered via the vaginal route spontaneously and died 37 days later. The autosite had cardiac hypertrophy with VSD, PDA, and PFO and an omphalocele. The liver was partly conjoined. There was a small swelling at the lower part of the autosite's bifurcated sternum to which the parasitic left pelvis and lower extremity were attached. On the second day after birth, parasitectomy was performed. The amputated specimens consisted of a small left pelvic girdle with a free extremity, a scrotal mass, nipplelike structures and two small protuberances externally. The liver, intestines, two testes, one kidney, one ureter, and the bladder were contained within the omphalocele. Although all of the organs and external structures of the parasite had abnormal histopathological findings, differentiated muscle fibers and submucous and/or myenteric plexus were observed.