Feeding ecology of two lanternfishes Diaphus garmani and Diaphus chrysorhynchus.

The feeding ecology of two dominant lanternfishes Diaphus garmani and Diaphus chrysorhynchus was studied in the continental slope region of the East China Sea, off western Kyushu (31-33° N; 128-130° E). Stomach contents of D. garmani were composed mainly of crustacean zooplankton, such as copepods, euphausiids, decapod larvae and amphipods, and also of appendicularians. Stomach contents of D. chrysorhynchus were composed mainly of crustacean zooplankton, cephalopods and fishes. Diel changes in stomach fullness indicated that D. garmani fed more actively at night than in the day. On the other hand, although feeding activity of D. chrysorhynchus did not change drastically between day and night, it tended to feed on large prey items in the benthopelagic zone during the day and on zooplankton in the epipelagic zone at night. Daily rations of food were estimated to be 2·54% of body dry mass for D. garmani, and 2·38% of body dry mass for D. chrysorhynchus.

Long-term effects of hepatocyte growth factor gene therapy in rat myocardial infarct model.

We investigated the long-term effects of human hepatocyte growth factor (HGF) gene therapy in a rat myocardial infarct model. Treatment adenovirus coexpressing the HGF therapeutic gene and the human sodium/iodide symporter (NIS) reporter gene or control adenovirus expressing the NIS gene alone were injected directly into the infarct border zone immediately after permanent coronary ligation in rats (n=6 each). A uniform disease state was confirmed in the acute phase in terms of impaired left ventricular (LV) function by cine magnetic resonance imaging (MRI), large infarct extent by (99m)Tc-tetrofosmin single-photon emission computed tomography (SPECT) and successful gene transfer and expression by (99m)TcO(4)(-) SPECT. After a 10-week follow-up, repeated cine MRI demonstrated no significant difference in the LV ejection fraction between the time points or groups, but a significantly increased end-diastolic volume from the acute to the chronic phase without a significant difference between the groups. Capillary density was significantly higher in the treatment group, whereas arteriole density remained unchanged. Two-photon excitation fluorescence microscopy revealed extremely thin capillaries (2-5 µm), and their irregular networks increased in the infarct border zone of the treated myocardium. Our results indicated that single HGF gene therapy alone induced an immature and irregular microvasculature.

Simultaneous evaluation of long-lasting knee synovitis in patients undergoing arthroplasty by power Doppler ultrasonography and contrast-enhanced MRI in comparison with histopathology.

OBJECTIVES: We simultaneously assessed ultrasonography (US) and magnetic resonance imaging (MRI) in comparison with histopathological changes in the knee joints of long-lasting arthritis patients. METHODS: We studied 15 patients with rheumatoid arthritis and 5 patients with osteoarthritis, who underwent total knee arthroplasty. On the day before surgery, the joints were examined by US and contrast-enhanced MRI. In US, synovitis was graded with 0-3 grey scale (GSUS) and power Doppler (PDUS). In MRI, synovitis was graded according to OMERACT-RAMRIS (grade 0-3). Synovial tissue samples were obtained during arthroplasty and evaluated on the basis of inflammatory cell infiltrates (grade 0-3), synovial lining layer thickness (grade 0-3) and vascularity (grade 0-3). RESULTS: Positive findings of PDUS and contrast-enhanced MRI were 45% and 85% of 20 operated joints, respectively. GSUS, PDUS and MRI synovitis were well correlated with overall histopathological grades of synovitis (Spearman correlation coefficients 0.48, 0.84 and 0.48, p<0.05, p<0.01 and p<0.05, respectively). Moreover, positive PDUS findings were closely associated with all pathological comportments of synovitis including inflammatory cell infiltrates, synovial lining layer thickness and vascularity. CONCLUSIONS: The present study revealed that positive PDUS findings more faithfully illustrated active synovitis than MRI, whereas contrast-enhanced MRI was more sensitive in detecting synovitis in patients with long-lasting arthritis. It is important to understand distinct features of the both modalities for clinical assessment of chronic joint diseases.

A mechanism responsible for the induction of H-2 restricted second order suppressor T cells.

The mechanism by which I-J restrictions were imposed on second-order suppressor cells (Ts2) was analyzed. The induction of Ts2 cells requires presentation of an inducer suppressor factor by a specialized population of factor-presenting cells. The I-J phenotype of this factor-presenting population controls the H-2 restriction of the Ts2 cells. The splenic cells responsible for presenting inducer factor appear to be of macrophage or dendritic cell lineage. Several homologies exist between the mechanism responsible for the induction of H-2-restricted suppressor and helper T cells. Thus, the I region products on specialized presenting cells determine the specificity and genetic restrictions of the T cell. In an H-2 heterozygous F1 animal, two distinct populations of cells can be induced, one specific for each parental H-2 heplotype. Furthermore, the data suggest that the suppressor cells also bear receptors for self H-2 products. The ramifications of these observations for the suppressor cell cascade are discussed.

The role of I-J and Igh determinants on F1-derived suppressor factor in controlling restriction specificity.

In the 4-hydroxy-3-nitrophenyl acetyl (NP) contact sensitivity system, the activity of third-order suppressor cells and their factors is restricted by H-2(I-J) and Igh linked genes. The present report analyzes the specificity of NP-specific Ts3 cells and factors derived from H-2 and Igh heterozygous (B6 X C3H)F1 mice. Two approaches were used. First, heterogeneous populations of F1 Ts3 cells were activated in vitro and then assayed in Ts3-depleted recipients which carried different combinations of H-2 and Igh alleles. The second approach was to hybridize the Ts3 cells and analyze the specificity of the F1-derived TsF3. The combined data demonstrated four functionally distinct populations of Ts3 cells. The activity of each population was restricted by a particular combination of H-2 and Igh haplotypes. Thus, Ts3 cells derived from F1 donors can demonstrate an apparent scrambling of H-2 and Igh restriction specificities. There was functional allelic exclusion of the H-2(I-J) and Igh determinants expressed on (B6 X C3H)F1 hybridoma-derived TsF3. Thus, TsF3 from each cloned hybridoma line expressed only one set of I-J and Igh determinants. Furthermore, there was a complete correlation between the I-J and Igh linked determinants expressed on TsF3 and the restriction specificity. In view of the recent findings on the molecular biology of the I-J region, an alternative interpretation of the role of I-J determinants on suppressor cells and factors is offered.

Visualization of in vivo electroporation-mediated transgene expression in experimental tumors by optical and magnetic resonance imaging.

In vivo electroporation (EP) is an efficient method for effective gene transfer and is highly expected for application in anticancer gene therapy. Non-invasive monitoring of gene transfer/expression is critical for optimal gene therapy. Here we report in vivo optical and high-field magnetic resonance imaging (MRI) of EP-mediated transgene expression in a tumor model. Initially, we observed spatio-temporal change in in vivo EP-mediated transgene expression by optical imaging using red fluorescence protein (RFP) as a reporter gene. Next, we constructed a dual-reporter plasmid carrying a gene-encoding MRI reporter ferritin heavy chain and RFP gene to visualize the intratumoral transgene expression by dual modality. Cells transfected with this plasmid showed lower signal intensity on in vitro T(2)-weighted cellular MRI and quantitatively increased the transverse relaxation rate (1/T(2)) compared with control cells. After conducting in vivo EP in an experimental tumor, the plasmid-injected region showed both fluorescent emissions in optical imaging and detectably lowered signal on T(2)-weighted MRI. The correlative immunohistological findings confirmed that both the reporter transgenes were co-expressed in this region. Thus, our strategy provides a platform for evaluating EP-mediated cancer gene therapy easily and safely without administering contrast agent or substrate.

Biphasic aspect of sexually dimorphic ontogenetic trajectory of gyrification in the ferret cerebral cortex.

The present study characterized quantitatively sexual dimorphic development of gyrification by MRI-based morphometry. High spatial-resolution 3D MR images (using RARE sequence with short TR and minimum TE setting) were acquired from fixed brain of male and female ferrets at postnatal days (PDs) 4-90 using 7-tesla preclinical MRI system. The gyrification index was evaluated either throughout the cerebral cortex (global GI) or in representative primary sulci (sulcal GI). The global GI increased linearly from PD 4, and reached a peak at PD 42, marking 1.486±0.018 in males and 1.460±0.010 in females, respectively. Sexual difference was obtained by greater global GI in males than in females on PD 21 and thereafter. Rostrocaudal GI distribution revealed an overall male-over-female sulcal infolding throughout the cortex on PD 21. Then, an adult pattern of sexually dimorphic cortical convolution was achieved so that gyrification in the temporo-parieto-occipital region was more progressive in males than in females on PD 42, and slightly extended posteriorly in males until PD 90. In the sulcal GI, sulcus-specific male-over-female GI was revealed in the rhinal fissure, and presylvian sulcus on PD 42, and additionally in the coronal, splenial, lateral, and caudal suprasylvian sulci on PD 90. The current results suggest that age-related sexual dimorphism of the gyrification was biphasic in the ferret cortex. A male-over-female gyrification was allometric by PD 21, and was thereafter specific to primary sulci located on phylogenetically newer multimodal cortical regions.

MRI of cerebral micro-vascular flow patterns: A multi-direction diffusion-weighted ASL approach.

The study and clinical assessment of brain disease is currently hindered by a lack of non-invasive methods for the detailed and accurate evaluation of cerebral vascular pathology. Angiography can detect aberrant flow in larger feeding arteries/arterioles but cannot resolve the micro-vascular network. Small vessels are a key site of vascular pathology that can lead to haemorrhage and infarction, which may in turn trigger or exacerbate neurodegenerative processes. In this study, we describe a method to investigate microvascular flow anisotropy using a hybrid arterial spin labelling and multi-direction diffusion-weighted MRI sequence. We present evidence that the technique is sensitive to the mean/predominant direction of microvascular flow in localised regions of the rat cortex. The data provide proof of principle for a novel and non-invasive imaging tool to investigate cerebral micro-vascular flow patterns in healthy and disease states.

Clinical implications of expression of interleukin 8 related to angiogenesis in uterine cervical cancers.

There was a significant correlation between microvessel counts and interleukin (IL)-8 levels and between infiltrated macrophage counts and IL-8 levels in uterine cervical cancers. Immunohistochemical staining revealed that the localization of IL-8 was similar to that of CD68 for macrophages. The prognosis of the 20 patients with high IL-8 (>1000 pg/mg protein) in uterine cervical cancers was extremely poor, whereas the 24-month survival rate of the other 60 patients with low IL-8 (<1000 pg/mg protein) was 67%. Therefore, this indicates that IL-8 might be a prognostic indicator as an angiogenic factor supplied from macrophages within and around the tumor.

The value of platelet-derived endothelial cell growth factor as a novel predictor of advancement of uterine cervical cancers.

Serum platelet-derived endothelial cell growth factor (PD-ECGF) in patients with uterine cervical cancers revealed a significantly positive correlation with clinical stage and tumor size and with the advancement indicators lymph node metastasis, parametrial involvement, and vessel permeation in both squamous cell carcinomas and adenocarcinomas. The prognosis of the patients with high serum PD-ECGF was extremely poor, whereas the 36-month survival rate of the other patients with low serum PD-ECGF was 81.3% in squamous cell carcinomas and 80.0% in adenocarcinomas. Our data indicate that serum PD-ECGF levels reflect the status of advancement of uterine cervical cancers and thus may be recognized as a novel tumor marker for both squamous cell carcinomas and adenocarcinomas of the uterine cervix.

Genetic control by I-A subregion in H-2 complex of incidence of streptozocin-induced autoimmune diabetes in mice.

An experimental autoimmune diabetes in mice characterized by delayed-onset hyperglycemia with lymphocytic infiltrations of the pancreatic islets can be induced by multiple administrations of low doses of streptozocin (STZ). We report on the influence of the MHC (H-2 complex) on this autoimmune diabetes by comparing the susceptibilities of various congenic and recombinant strains with a B10 background. In congenic strains, C57BL/10 (H-2b) and B10.BR (H-2k) mice showed a high incidence of diabetes, whereas B10.D2 (H-2d) and B10.S (H-2s) mice showed a low incidence. Therefore, we suggest that the H-2 complex influences diabetes susceptibility and that both b and k are high-susceptibility alleles, whereas d and s are low-susceptibility alleles. In recombinant strains, those with the same haplotypes on the K, E, S, and D subregions of the H-2 complex showed undefined (high and low) susceptibilities, indicating that the diabetes-susceptibility genes are located outside these loci. Strains possessing I-Ab or I-Ak gene products (C57BL/10, B10.BR, B10.TL, B10.A, and B10.A(2R] showed high incidences, whereas strains possessing I-Ad or I-As (B10.D2, B10.S, B10.S(7R), B10.S(9R), and B10.GD) showed low incidences. In addition, administration of anti-I-A monoclonal antibody prevented the manifestation of diabetes in STZ-administered mice. Passive transfer of STZ-administered T lymphocytes to mice given minute doses of STZ induced significant hyperglycemia. This successful transfer was only observed in H-2-compatible mice. Thus, we conclude that one gene coding for susceptibility to this experimental diabetes was located in the I-A subregion within the H-2 complex.

Platelet-dependent thrombin generation in patients with hyperlipidemia.

OBJECTIVES: We evaluated coagulability as determined by platelet-dependent thrombin generation in hypercholesterolemic patients before and after treatment with pravastatin and in hypertriglyceridemic patients to investigate the usefulness of coagulability as an index of atherosclerosis and to determine the importance of treating hyperlipidemia. BACKGROUND: An understanding of the interaction between platelets and the plasma coagulation system is important for clarifying the mechanism of the procoagulant process. METHODS: We assessed coagulability in 58 patients with hypercholesterolemia (serum total cholesterol level > or = 220 mg/dl, age 56.5 +/- 1.5 years [mean +/- SEM]), 37 patients with hypertriglyceridemia (serum triglyceride level > or = 200 mg/dl, age 59.5 +/- 1.7 years), 13 patients with hypercholesterolemia plus hypertriglyceridemia (age 51.4 +/- 3.1 years) and 75 normal subjects (age 52.2 +/- 1.7 years). We also studied platelet-dependent thrombin generation in patients with hypercholesterolemia before and after treatment with pravastatin. Calcium chloride was added to 0.5 ml of platelet-rich plasma (150 x 10(9)/liter) to initiate coagulation. Ten microliters of the sample was transferred into 90 microliters of 3.8% sodium citrate at 10-min intervals for 30 min. A chromogenic substrate, S-2238, was added to each sample, and absorbance was measured spectrophotometrically at a wavelength of 405 nm to determine thrombin generation. RESULTS: Platelet-dependent thrombin generation was increased in patients with hypercholesterolemia and patients with hypercholesterolemia plus hypertriglyceridemia (p < 0.01) compared with patients with hypertriglyceridemia and control subjects. Treatment with pravastatin normalized thrombin generation. CONCLUSIONS: Hypercholesterolemia, but not hypertriglyceridemia, was associated with increased platelet-dependent thrombin generation. Pravastatin normalized the generation of thrombin.

Platelet-dependent thrombin generation in patients with diabetes mellitus: effects of glycemic control on coagulability in diabetes.

OBJECTIVES: This study sought to assess the usefulness of platelet-dependent thrombin generation as an index of coagulability in diabetes and to determine the effect of glycemic control on coagulability in diabetes. BACKGROUND: It is important to investigate the interaction of platelets and the coagulation factors to clarify the processes of the coagulation system in detail. METHODS: Platelet-rich plasma (150 X 10(9)/liter), 0.5 ml, was prepared, and 40 mmol/liter of calcium chloride was added to initiate clotting. S-2238 was added to each sample in a microtiter plate every 10 min, and the absorbance of the released color product at 2 min was measured spectrophotometrically at a wavelength of 405 nm using a microtiter plate reader as thrombin generation. We measured the platelet-independent thrombin generation in patients with non-insulin-dependent diabetes mellitus grouped according to glycemic control. RESULTS: Platelet-dependent thrombin generation at 30 min after calcium chloride addition was significantly higher in 23 patients with poorly glycemic-controlled non-insulin-dependent diabetes mellitus without complications, such as diabetic retinopathy, nephropathy and neuropathy (hemoglobin [Hb] A1c >/= 9.0%) than in 46 healthy normal subjects (448 +/- 75 vs. 165 +/- 28 mU/min, p < 0.001). Thrombin generation in 31 well controlled diabetic patients without complications (Hb A1c < 9.0%) was intermediate (240 +/- 72 mU/min) between those of the poorly controlled group and healthy normal subjects. Platelet-poor plasma from diabetic patients increased platelet-dependent thrombin generation in normal subjects. CONCLUSIONS: Coagulability is evidently enhanced in patients with non-insulin-dependent diabetes mellitus compared with that in healthy normal subjects on the basis of assessments of the platelet-dependent thrombin generation, and good glycemic control may help to correct a hypercoagulable state in diabetic patients.

Primary care in the treatment of functional gastrointestinal symptoms in Japan: prescription preferences and impression of results.

BACKGROUND: Functional gastrointestinal (GI) disorders are common in primary care. However, proper pharmacological approaches have not yet been established. The reason for a lack of proper approaches may be attributable to the lack in clarity of their pathogenesis and pathophysiology. Meta-analysis of pharmacological approaches to functional GI disorders failed to identify the solid cluster of patients' symptoms. AIM: The aim of this study is to assess the perspective of primary care doctors concerning prescriptions for functional GI symptoms, evaluate the efficacy of the drugs prescribed, and the need for medication for these symptoms. METHOD: Questionnaires were sent to primary care doctors, and a total of 149 responses were obtained. Efficacy of each medication was evaluated by the number of doctors favouring the category, and the respective impressions of prescriptions given. RESULTS: Symptoms of heartburn were well controlled by anti-secretory drugs (H2RAs and PPIs), while appetite loss and abdominal gurgling were not controlled by any medications. CONCLUSIONS: This survey reveals differences in need for various prescription drugs in functional GI symptoms.

Erythroid progenitors in tumor-bearing mice: erythroid inhibitory factors produced by adenocarcinoma-755.

Adenocarcinoma-755-bearing C57BL/6 mice developed anemia with the growth of the tumors. The numbers of granuloid and monocytoid progenitors (colony-forming unit in culture, CFU-C) of the bone marrow and spleen and the peripheral blood WBC counts increased in tumor-bearing mice. The erythroid progenitors (erythroid colony-forming units, CFU-E; erythroid burst-forming units, BFU-E) of the bone marrow showed a marked decrease, overcoming their increase in the spleen in tumor-bearing mice. Fractionation of conditioned medium of tumor cells led to the isolation of a protein of 80-kd molecular weight that stimulated murine CFU-C growth and inhibited the CFU-E and BFU-E growth in a dose-dependent manner. These results indicate that erythroid inhibitory factors produced by tumors exist in tumor-bearing mice. Erythroid inhibitory factors might also exist in non-hematological malignancies of humans, and they might be one of the mechanisms of anemia.

Effects of rhIL-1 alpha, rhIL-1 beta, and rhIL-1 receptor antagonist on erythroid progenitors (CFU-E and BFU-E) in human bone marrow.

Interleukin-1 (IL-1) is known to promote the production of colony-stimulating factor (CSF) and to possess the ability to protect the bone marrow suppression in granulocyte-macrophage (GM) series associated with radiotherapy or chemotherapy. There are conflicting reports concerning the action of IL-1 on erythroid progenitors, however, and no consensus has been established. In the present study, the influences of recombinant human IL-1 alpha (rhIL-1 alpha), rhIL-1 beta, and rhIL-1 receptor antagonist (rhIL-1ra) on erythroid progenitors (colony-forming units-erythroid, CFU-E; and burst-forming units-erythroid, BFU-E) in human bone marrow were studied. rhIL-1 alpha and rhIL-1 beta (1-1000 pg/mL) enhanced the CFU-E and BFU-E growth in human nonadherent (NA) bone marrow cells. rhIL-1 alpha and rhIL-1 beta also stimulated the formation of CFU-E and BFU-E in the NA and T cell-depleted bone marrow fraction. Moreover, rhIL-1 alpha and rhIL-1 beta enhanced the CFU-E and BFU-E in the CD34+ bone marrow cell fraction. These data and the results of limiting dilution analysis indicate that the stimulatory effect of IL-1 may consist of direct actions on erythroid progenitors. The enhancing effect of rhIL-1 alpha and rhIL-1 beta on erythroid progenitors was inhibited by rhIL-1ra. These data suggest that the stimulatory effect of IL-1 on CFU-E and BFU-E is mediated via the IL-1 receptor.

Effects of ubenimex on erythroid progenitors (CFU-E and BFU-E) in human bone marrow.

Ubenimex (UBX, bestatin) is known to be an immunomodulator and host-mediated antineoplastic agent. Effects of UBX on human bone marrow erythroid progenitors (erythroid colony-forming units, CFU-E; and erythroid burst-forming units, BFU-E) were investigated in vitro. UBX enhanced CFU-E and BFU-E growth in the nonseparated bone marrow mononuclear cell fraction at concentrations from 0.005 to 5 micrograms/ml. The enhancements of CFU-E and BFU-E were independent of the concentration of erythropoietin added to culture system. In the T-cell-depleted bone marrow fraction, UBX also increased CFU-E and BFU-E growth, but it failed to stimulate these cells in the nonphagocytic and nonadherent bone marrow fraction. These findings indicate that UBX may stimulate erythroid progenitors mediated through monocytes and macrophages.

Suppressor T cells in mice made unresponsive to skin allografts.

We have induced suppressor T cells (Ts), capable of delaying allogeneic skin graft rejection, through the intravenous administration of allogeneic spleen cells under normal conditions. H-2 and non-H-2 incompatibility between recipient mice and donor skins induced strong graft rejection. However, when the Ts were transferred into recipient mice, the mean survival time was prolonged for every combination between recipient mice and donor skin. Studies using several strains of congenic mice revealed the antigen specificity of these Ts. Treatment with monoclonal anti-Lyt-2.2 or anti-Thy-1.2 antibody and complement abolished the suppression shown by the Ts of skin graft rejection. The suppression induced by these Ts, however, was resistant to treatment with monoclonal anti-Lyt-1.2, anti-L3T4, or anti-I-A antibody and complement. These results showed that the Ts were Lyt-1-2+, L3T4-, Ia-, T cells.

Genetic control of nickel sulfate delayed-type hypersensitivity.

Genetic control of delayed-type hypersensitivity (DTH) to nickel sulfate antigen was studied using various strains on inbred mice. Mice with class II Ad,k,s haplotypes showed a high magnitude of response, whereas those with class II Ab,f were low responders. These results indicate that nickel sulfate DTH may be influenced by the I-A region of H-2. Transfer experiments revealed that these responses might be mediated by L3T4+, Lyt-2- T cells.

Sequencing and cloning of the cDNA of guinea pig eosinophil major basic protein.

Major basic protein (MBP) purified from guinea pig eosinophils elicited histamine release from rat peritoneal mast cells at concentrations higher than 3 micrograms/ml both in the presence and in the absence of extracellular Ca2+. After reverse-phase high-performance liquid chromatography, it was revealed that MBP was composed of two different proteins with quite similar molecular weights and pI values, although the amino acid compositions were slightly different. The partial amino acid sequence of one of these MBPs was determined and the primers for the polymerase chain reaction (PCR) were synthesized according to the partial amino acid sequence. Using these primers and the cDNAs obtained from guinea pig eosinophils, the PCR was carried out in order to synthesize the hybridization probe of MBP for screening the cDNA library. After screening with 8 x 10(5) clones, a positive clone, which encoded a full length of pre-proMBP, was obtained. According to the sequencing data of this clone, it was revealed that pre-proMBP was composed of 3 domains; signal peptide, acidic domain and mature MBP. The predicted pI value of mature MBP was 11.7, though that of proMBP was 7.8. The homology in the amino acid sequence between guinea pig proMBP and human proMBP was 49.4%, while guinea pig mature MBP was more homologous (58%) to human mature MBP.