TRIM27 expression is associated with poor prognosis in sinonasal mucosal melanoma.

BACKGROUND: Tripartite motif-containing 27 (TRIM27) has been implicated in the progression of various cancers. However, the role of TRIM27 in sinonasal mucosal melanoma (SNMM) remains poorly understood. MATERIALS & METHODS: We retrospectively examined 28 patients with SNMM treated with between 2003 and 2021. We undertook immunohistochemical analysis of TRIM27, Ki-67, and p-Akt1 expression in SNMM tissues. We also investigated the relationship between TRIM27 expression and clinical characteristics, prognosis, Ki-67 as a tumor growth potential marker, and p-Akt1 as one of the prognostic factors in mucosal melanoma. RESULTS: TRIM27 expression was significantly higher in T4 disease than in T3 disease and was higher in stage IV than in stage III. Patients with high-TRIM27 SNMM had a significantly poorer prognosis in terms of overall survival (OS) and disease-free survival.There was also a significantly higher rate of distant metastasis. Univariate analysis for OS revealed that TRIM27 and T classification were significant poor prognostic factors. In addition, the Ki-67 positive score and the p-Akt1 total staining score were significantly higher in the high-TRIM27 group than in the low-TRIM27 group. CONCLUSIONS: High TRIM27 expression in SNMM was associated with advanced T classification, poor prognosis and distant metastasis. We suggest that TRIM27 has potential as a novel biomarker for prognosis in SNMM.

Characterization of primary structure and tissue expression profile of the chicken apical sodium-dependent bile acid transporter mRNA.

The ileal apical sodium-dependent bile acid cotransporter (ASBT) plays an essential role in the absorption of bile acids from intestinal lumina. ASBT cDNA has been cloned from mammalian and fish species, and the primary structure of the protein and expression properties of the mRNA have been characterized. In this study, we identified chicken ASBT mRNA by cDNA cloning. Chicken ASBT cDNA consisted of 91 bp of the 5'-untranslated region, 1,083 bp of the coding region, and 1,896 bp of the 3'-untranslated region. The cDNA encoded a protein of 360 amino acids showing significant sequence identity with mammalian and fish ASBT. The amino acid residues known to participate in the functions of mammalian ASBT were conserved in chicken ASBT. Real-time polymerase chain reaction analysis revealed that chicken ASBT mRNA was expressed at markedly higher levels in the ileum and proximal colon/rectum, relatively lower levels in the kidney, and very low levels in the jejunum and cecum. Expression levels in the ileum markedly increased after hatching, reached the highest levels on day 7 posthatching, and then decreased to adult levels. A similar expression pattern was observed in the proximal colon/rectum except for the significant decrease from day 7 posthatching to day 21 posthatching. These results suggest that chicken ASBT functions as a bile acid transporter in the ileum and proximal colon/rectum, particularly during the early posthatching period.

Changes in expression levels of neurotensin precursor and receptor mRNA in chicken intestinal tissues and liver during late embryonic and early posthatching development.

Neurotensin is a tridecapeptide that has multiple functions as a neurotransmitter and as a circulating hormone. Neurotensin and its related peptide, LANT6, have been isolated in the chicken, but the mRNA encoding these peptides has not been identified. In this study, we first cloned the cDNA for the chicken neurotensin precursor mRNA from the duodenum and characterized its primary structure and then investigated tissue expression patterns of neurotensin precursor and receptor mRNA. The cDNA encoded a protein of 495 amino acids that contains the sequences of chicken neurotensin and LANT6 in the C-terminal region. Real-time PCR analysis showed that the neurotensin precursor mRNA is preferentially expressed in intestinal tissues, such as the duodenum, jejunum, ileum, and colon/rectum, with temporal increases during the hatching period. The expression levels of neurotensin receptor 1 mRNA were relatively higher during the late embryonic period compared with the posthatching period in the duodenum and jejunum, whereas the expression levels were higher in the colon/rectum during the posthatching period. In the liver, the expression levels of neurotensin receptor 1 were markedly increased during the early posthatching period. These results suggest that chicken neurotensin is involved in the regulation of gastrointestinal and hepatic functions, especially during the hatching period.

Primary structure and tissue distribution of GPR39 messenger ribonucleic acid in Japanese quail, Coturnix japonica.

It has been found that GPR39 is an orphan receptor that belongs to the family of G protein-coupled receptors. In mammals, GPR39 has been shown to be involved in the regulation of gastrointestinal and metabolic function. In this study, we performed cDNA cloning for GPR39 in Japanese quail and characterized the tissue expression profiles of its mRNA. The cDNA encoded 462 amino acids, showing very high sequence homology to chicken GPR39 (95.5%) and moderate homology to mouse (64.7%), rat (63.7%), and human (59.9%) GPR39. Real-time PCR analysis revealed that GPR39 mRNA is expressed at high levels in the digestive tissues such as stomach, duodenum, jejunum, ileum, cecum, and colon and rectum and at moderate levels in the oviduct including infundibulum, magnum, isthmus, and uterus. These findings suggest that GPR39 may be involved in gastrointestinal and oviductal functions in Japanese quail.

Characterization of primary structure and post-hatching increase in chicken cytosolic acetoacetyl-coA thiolase in the liver.

Acetoacetyl-CoA thiolase (EC 2.3.1.9) catalyzes the cleavage of acetoacetyl-CoA into acetyl-CoA and its reverse reaction, the synthesis of acetoacetyl-CoA. Cytosolic acetoacetyl-CoA thiolase ( CT: ) is a key enzyme in the initial step of the cholesterol synthesis pathway. In the present study, we characterized the amino acid sequence of chicken CT and the tissue distribution of its mRNA and protein, together with their developmental changes in the liver. The amino acid sequence encoded by the nucleotide sequence of chicken CT cDNA showed a higher overall identity with those of human (74.3%) and rat (74.6%) CTs. Amino acid residues known to participate in enzymatic activity in human CT are conserved in chicken CT. Real-time PCR analysis revealed the expression of CT mRNA in the liver, kidney, adrenal gland, jejunum and ovary of adult hens, with higher levels in the liver, kidney, adrenal gland and ovary. Western blot analysis detected an immunoreactive protein of 41 kDa from cytoplasmic fraction but not particulate fractions of adult chicken liver. The immunoreactive protein was detected in all the tissues. The mRNA levels in the liver rapidly increased after hatching, with a maximum on d 5 post-hatching, after which they gradually decreased to adult levels. A similar change was observed in the protein levels. The increase in transcription and protein synthesis of CT suggests that the synthetic pathway of cholesterol from acetyl-CoA produced by CT replaces the hydrolysis of accumulated cholesteryl ester in the liver, in response to a change in the nutrient source from the lipid-rich yolk to a lower-lipid diet during the early post-hatching period.

The clinical features and treatment of arteriosclerosis obliterans with diabetes.

The objective of this study was to clarify the differences, if any, in the clinical features between diabetic and nondiabetic patients with arteriosclerosis obliterans (ASO) and to select the optimal treatment for diabetic patients with ASO. The 171 patients with ASO studied were classified into nondiabetic and diabetic groups. Each group was subdivided into an intermittent claudication (IC) group and ulcer and necrosis (ULC) group. The frequency of complications with cardiac and cerebral vascular diseases and risk factors of arteriosclerosis were analyzed. Ankle and brachial blood pressure and ankle/brachial pressure index (API) were measured, and blood rheological parameters of filterability using Nuclepore filter membrane and viscosity of whole blood and plasma were measured. Three indexes of walking distance were measured by our ASO-Treadmill protocol to evaluate quantitatively the effect of treatment. There were 95 diabetic patients with ASO and 76 nondiabetic patients. Of the nondiabetic patients, 81 had IC and 14 had ULC, and of the diabetic patients, 63 had IC and 13 had ULC. The diabetic group showed more frequent complications with coronary heart disease (56.5 vs. 25.6%), but the two groups showed the same frequency of cerebrovascular diseases (30%). The diabetic ULC subgroup showed higher fasting plasma glucose than the diabetic IC subgroup. The API of the ULC subgroup was significantly lower than that of the IC subgroup in the nondiabetic patients, whereas that of the ULC subgroup was not significantly lower than that of the IC subgroup in the diabetic patients. Stenotic lesions of arteriography in both the nondiabetic and diabetic ULC subgroups demonstrated a tendency toward multisegmental and below-knee lesions compared with the two IC subgroups. For blood rheology-related factors, the diabetic ASO subgroup demonstrated a significantly elevated fibrinogen level compared with the normal control value, for patients of average age. After walking exercise treatment, a significant increase in the walking distance was obtained. After treatment with Cilostazol, prostaglandin I2 analog, and LDL apheresis, the rheological indexes were significantly improved, while the API did not change. We conclude that therapeutic improvement of blood rheological properties would be effective for prolongation and improvement of the quality of life for diabetic patients with ASO.

Effects of elastase on the stiffness and elastic properties of arterial walls in cholesterol-fed rabbits.

The effect of cholesterol feeding and elastase administration on the arterial stiffness and elastic properties was investigated in rabbits by pressure-diameter tests on excised thoracic aortas, common carotid and femoral arteries. Rabbits in groups RA and RB were fed 1% cholesterol diet for 14 weeks. Groups RC and RD were fed the cholesterol diet for the first 7 weeks, and then given regular chow for the last 7 weeks. In addition, rabbits in groups RB and RC were administered elastase daily for the last 7 weeks. Rabbits in group RE were fed the regular chow for 14 weeks and served as the control group. The arterial stiffness and the elastic modulus of wall material in the RA rabbits were generally higher than those in the control rabbits. Administration of elastase and/or feeding of the regression diet decreased the wall stiffness and elastic modulus significantly, although the effect of elastase did not appear clearly when used in combination with the regression diet. The cholesterol feeding decreased the ratio of thickness to wall radius, whereas the ratio was more or less increased by the elastase administration and/or the regression diet.

Three missense mutations in the protein C heavy chain causing type I and type II protein C deficiency.

We have studied the molecular basis of protein C deficiency in three families with a history of thromboembolic disease. An approximately 50% reduction in both functional and immunologic levels of protein C was detected in the plasma from two unrelated patients, designated protein C Osaka 1 and protein C Osaka 2. An approximately 50% reduction in functional level but normal immunologic level of protein C was detected in plasma from a third patient, designated protein C Osaka 3. DNA sequencing of the amplified DNA revealed one missense mutation in each case. Additional mutations in the coding sequence were excluded by DNA sequencing of all protein C exons. We identified a C-to-T change at nucleotide number 6,218 of the protein C gene in protein C Osaka 1. This results in the amino acid substitution of Arg-169 by Trp at the alpha-thrombin cleavage site. In protein C Osaka 2, a G-to-A change at nucleotide number 8,807 was identified leading to the amino acid substitution of Met-364 by Ile in the protease domain. This substitution may impair the synthesis or stability of protein C Osaka 2. In protein C Osaka 3, a G-to-C change at nucleotide number 8,868 was identified. This results in substitution of Gly-385 by Arg in the protease domain. Based on these, it was concluded that Arg-169-to-Trp mutation and Met-364-to-Ile mutation cause type I protein C deficiency and Gly-385-to-Arg mutation causes type II deficiency.

Immunoreactive transferrin receptor in sera of pregnant women.

Immunoreactive transferrin receptors in sera of 90 pregnant women of various gestational periods were investigated. The mean concentrations of the serum transferrin receptor in normal males and females were 251 +/- 94 ng/ml and 256 +/- 99 ng/ml, respectively. Serum transferrin receptor levels in pregnant women did not show a significant increase in the early stage of gestation. However, a rapid elevation of the mean concentration of transferrin receptor was observed after 20 weeks of pregnancy. The apparent increase with gestation period suggests that this immunoreactive receptor in the sera of pregnant women is derived from the placental syncytiotrophoblast and reflects the activity of maternofetal iron transport.

Successful treatment of a VIPoma by continuous subcutaneous infusion of somatostatin analogue (SMS 201-995).

A case with WDHA syndrome due to VIPoma is reported. Injection of somatostatin analogue SMS 201-995 was followed by prompt suppression of vasoactive intestinal polypeptide levels (VIP), decreased stool volume, and restoration of the serum potassium concentration to normal. Long-term treatment with SMS 201-995 for up to 20 weeks produced excellent clinical control and a decrease in tumour size. No adverse effects were noted except for localized pain at the site of injection. This was overcome by using a continuous subcutaneous infusion pump which also enabled the effective daily dosage to be reduced and thereby adverse reactions to be avoided.

Circulating transferrin receptor in acute leukemias.

Serum transferrin receptor (s-TR) levels in acute leukemia patients were measured by a recently developed sandwich radioimmunoassay. The mean s-TR level for normal subjects (n = 205) was 246 +/- 79 (mean +/- 1 SD) ng/ml. The values for patients with untreated acute myelocytic leukemia (AML, n = 18) and untreated acute lymphocytic leukemia (ALL, n = 14) were 398 +/- 175 ng/ml and 479 +/- 176 ng/ml, respectively, both of which were significantly higher than those for the normal subjects (AML, p < 0.02; ALL, p < 0.05). When complete remission was achieved with initial remission induction therapy, s-TR decreased to 262 +/- 47 ng/ml (n = 22, 12 AML and 10 ALL), returning to normal levels. There was a good correlation between s-TR levels and the number of leukemic cells in peripheral blood (r = 0.743, n = 32, p < 0.01). In two patients with AML, serial changes of s-TR values and numbers of blast cells in the peripheral blood and bone marrow occurred in a parallel manner. If, therefore, this assay for s-TR can be made more sensitive, it may become useful for assessing acute leukemia activity and for monitoring the effects of therapy.

Blood rheology in the endotoxin-administered rabbits.

The effects of endotoxin on blood rheology were studied in rabbits. Non-treated six rabbits (Group A) were used to obtain control data. Thirteen rabbits who were administered 0.1 mg of endotoxin three times at intervals of 3 days were divided into two groups; seven rabbits (Group B) were injected with 1 ml saline solution as the vehicle for endotoxin at 7 days after the final administration of endotoxin, while the remaining six rabbits (Group C) were administered endotoxin at 0.2 mg/kg on the same day. Blood was sampled from the femoral artery 120 min after the final treatment. Blood viscosity was measured at a shear rate of 150 s-1 at 37 degrees C using a cone-plate viscometer. The passage time for a 5% red blood cell suspension and that for plasma were determined by filtration; the former represents erythrocyte deformability while the latter is related to plasma fluidity. The hematocrit, whole blood viscosity and erythrocyte deformability did not show significant differences among these three groups. The ratio of hematocrit to whole blood viscosity is considered to be an index of oxygen delivery from the hemorheologic point. This index did not show significant difference either. A good correlation was observed between whole blood viscosity and hematocrit in Group A, but not in the endotoxin-treated groups. The plasma fluidity was lower in Groups B and C than in Group A. These data indicate that plasma fluidity and the hematocrit-viscosity relationship are affected in endotoxin-treated rabbits, although blood viscosity, erythrocyte deformability and oxygen delivery hardly changed.

Tissue distribution and pharmacological potential of SM-16896, a novel oestrogen-bisphosphonate hybrid compound.

Postmenopausal osteoporosis is caused mainly by a deficiency of oestrogen with rapid bone loss. To target oestrogen to the bone effectively, we have synthesized and evaluated the effects of a novel hybrid compound of oestrogen and bisphosphonate, SM-16896. The tissue distribution pattern and pharmacological potential are reported. Although the affinity for calf uterine oestrogen receptor was very low (IC50: 73.3 microM; 1/25000 of that of 17beta-oestradiol (2.84 nM)), SM-16896 showed oestrogenic activity. SM-16896 (1 microM) induced a 4.5-fold transcriptional activity in rat osteosarcoma UMR-106 cells compared with vehicle-treated control, when we used the expression vector for human oestrogen receptor and a CAT reporter plasmid containing an oestrogen-responsive element. The distribution of SM-16896 after a subcutaneous administration to 7-week-old female rats was examined by radioluminography using 3H-labelled SM-16896. At 30 min after the administration, significant radioactivity was detected in the bone. At 24 h after administration, a high level of radioactivity was detected in the bone, but in the uterus it was only at a background level. Daily subcutaneous administration of 0.5 mgkg(-1) SM-16896 for 12 weeks (five times per week) to 13-week-old ovariectomized rats suppressed the ovariectomized-induced reduction in bone mineral density. A bone mineral density ratio of 120% was maintained compared with sham-operated rats, whereas a relatively low suppression of uterine weight was observed (about 50% loss compared with sham-operated rats). In the same experiment, the implantation of a 17beta-oestradiol time-release pellet (0.25 mg/pellet/90 days) almost completely suppressed the reduction of both the bone mineral density and uterine tissue weight. It is likely that the effect of SM-16896 on bone was due to its oestrogenic activity, since 1.0 mgkg(-1) SM-18108, the bisphosphonate moiety of this compound, had no effect on bone in 7-week-old ovariectomized rats. The results suggest that SM-16896, a bisphosphonate-conjugated oestrogen, showed a preference profile in the uterus and bone due to its characteristic distribution pattern compared with the natural oestrogen analogue 17beta-oestradiol. Thus, bisphosphonate-conjugated oestrogens have the potential to improve patient compliance in oestrogen therapy by minimizing adverse effects and reducing the frequency of medication.

Inherited heterozygous protein C deficiency and dysfunctional protein S with recurrent venous thrombotic diseases: a study of three generations of a Japanese family.

We describe a rare occurrence of a family affected with venous thrombosis, exhibiting a protein C (PC) deficiency and dysfunctional protein S (PS). The propositus and his father developed recurrent venous-thrombosis. Their PC deficiency was characterized by low levels of both antigen and activity, and their dysfunctional PS was suggested by low PS activities despite the presence of normal free PS antigen. Over three generations, six family members had a PC deficiency, and three had both a PC deficiency and a dysfunctional PS. The mode of inheritance of PC deficiency appears to be autosomal dominant.

Microvascular blood is distributed more to venules than to arterioles in patients with Buerger's disease. Observation of bulbar conjunctiva by intravital microscope system.

BACKGROUND: It has been a matter of controversy whether abnormalities of organs other than extremities may be a clinical manifestation of Buerger's disease (thromboangiitis obliterans; TAO). In the present investigation, our aim was to quantitatively characterise the configuration of microvascular networks in bulbar conjunctiva, which is not affected apparently, in patients with thromboangiitis obliterans. METHODS: Nine men with thromboangiitis obliterans attended our hospital and nine male volunteers as normal controls were enrolled in this study. We observed and analysed the configuration of the network of a bulbar conjunctiva by use of intravital microscope system with computer assisted image processing functions. Microvessel density was defined as a summation of vessel length in a ROI area and tortuosity was evaluated by a ratio of vessel length to direct distance of both terminals. RESULTS: In the microcirculation of bulbar conjunctiva in thromboangiitis obliterans, arteriole diameter was significantly decreased and density of venules was significantly increased. Increased venular density was mainly explained by increased tortuosity of venules. CONCLUSIONS: Consequently, microvascular blood was distributed more to venules than to arterioles in patients with thromboangiitis obliterans. Venule/arteriole ratios of diameter, tortuosity and microvessel density may be useful parameters to characterize the configuration of microvascular networks in thromboangiitis obliterans.

Endogenous amino acids in budgerigars (Melopsittacus undulatus).

To determine the amount of endogenous amino acids in budgerigars (sekiseiinko, Melopsittacus undulatus), as the first step of evaluating amino acid requirements, three experiments were conducted. In Experiments 1 and 2, transit time of the feeds through the digestive tract in budgerigars was studied. The birds were given free access to a corn-based diet containing 50% barium sulfate in Experiment 1, and Japanese millets (hie, Echinochloa utilis) with 0.5 ml of 50% barium sulfate solution in Experiment 2. At intervals of 0.5 to 2 hr, roentgenography was conducted to identify the position of the barium sulfate in the digestive tract. The results showed that the transit time of pellets and millets in the digestive tract was within 24 and 27 hr, respectively. In Experiment 3, to determine the amount of endogenous amino acids in budgerigars, the excreta were collected for 24 hr, from 27 to 51 hr after fasting. The excreta were hydrolyzed with 6 N hydrochloric acid at 110 degrees C for 22 hr for the determination of amino acids by HPLC. The endogenous Ala, Arg, Leu, Lys, Phe and Val were estimated to be 34.9, 36.8, 18.5, 20.3, 15.3 and 18.9 micrograms/day/BW0.75, respectively, which were higher than those in adult roosters.

Study on reaction of subpapillary venous plexus in patients with arterial occlusive diseases of lower extremity by reflective spectrophotometry.

The reactions of the subpapillary venous plexus were observed by analyzed fractional blood volume of tissue (Vb) in patients with arterial occlusive diseases, Fontaine's stage I-II, using the reflective spectrophotometer. The sole skin of 11 normal subjects, 17 patients with arteriosclerosis obliterans (ASO), and 14 patients with thromboangiitis obliterans (TAO) were scanned by the light reflectance of four specially selected wavelengths, including isobestic points (586 and 569 nm) and peaks in the spectra of oxyhemoglobin and deoxyhemoglobin (577 and 560 nm) in referring to time. Sampling area was the subpapillary venous plexus histologically because the depth for receiving reflectance is about 1.0 mm from the sole skin surface. One series observations was divided into five steps depending on the modes of combination of the applied pressure to ankle cuff and tilting. The Kubelka-Munk equation was utilized to calculate blood volume (Vb). It is concluded that the patients with ASO might have a dysfunction, not of the subpapillary venous plexus, but of the venular valve, whereas patients with TAO might have a dysfunction of both the subpapillary venous plexus and venular valve.

Change of hematocrit and blood viscosity in cholesterol-fed rabbits.

In the cholesterol-fed rabbits, we observed that the whole blood viscosity was maintained at the normal level in spite of the decrease in hematocrit. This phenomenon suggests that there exists some visco-regulatory mechanism, and we could simulate it by a simple integration type model.

White blood cell adhesion to endothelium and rheological behavior in microvessels of overinflated frog's lung.

The flow behaviors of white blood cells (WBCs) in frog's pulmonary microvessels were recorded and analyzed by means of a microscope-TV camera system. When the flow velocity in arterioles was reduced to a level lower than 1 mm/sec by a moderate overinflation of the exposed lung, WBCs rolled on the endothelial surface, frequently came in contact with the capillary orifice and passed it quickly without deformation. The time length which was required for WBCs to pass through the capillary orifice was shorter than the time length for red blood cells. This observation suggested that WBCs were no hinderance to blood delivery from arterioles to the capillary network in the normal and moderate overinflation of the lung. However, when the lung was strongly overinflated and the center line flow velocity was reduced to 0.1 mm/sec, WBCs adhered to the endothelium in ten minutes. The adhering WBCs could not be detached by the recovery of the blood flow. It seemed probable that a large shear stress up to 100 to 200 dynes/cm2 was necessary to pull down the interaction between the adhering WBCs and the endothelium.

Effects of glucagon administration on microcirculation and blood rheology.

It has been reported that some rheologic abnormalities are associated with diabetes. However, it is difficult to realize the condition of microcirculation in men. We developed an intra-vital video-microscopic system (IVVMS) to observe microcirculation directly on the bulbar conjunctiva. Using this system, we investigated the effect of intravenous glucagon administration on microcirculatory parameters such as the internal diameter of venules, flow velocity and flow volumes in diabetic patients. Blood viscosity and rheology factors were also examined before and after glucagon administration. The time required for erythrocytes to pass through pores of 5 microm diameter, as well as whole blood viscosity were decreased significantly after glucagon administration. Flow volumes of venules were increased significantly by glucagon. Leukocyte counts, platelet counts and fibrinogen did not change after glucagon administration. However, plasma glucose, insulin, C-peptide and cyclic-AMP were increased. In conclusion, glucagon administration improved blood fluidity and modified the microcirculation in patients with diabetes mellitus.