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Biomedical subjects

H Harada

Publications and source records attributed to H Harada.

At least 235 records · Page 13Linked to original sources

Chronic pancreatitis: functional testing.

This article reviews the evolution of functional testing of the pancreas in Japan for the diagnosis and treatment of chronic pancreatitis (CP), contrasting the pre- with the postsecretin test (S test) era. In the pre-S test era, the diagnosis was based on symptoms, clinical findings, fasting serum diastase levels, and the vagostigmin- and ether-stimulation test unless morphologic evidence was available. The S test and CCK-pancreozymin (PZ) test (PS test) were introduced into Japan around 1963 and have been used as the gold standard of the exocrine pancreatic-function test. Through a series of attempts at standardization in 1971, 1985, and 1987, the method was standardized to collect duodenal juice for 60 min through a double- or triple-lumen tube after a bolus or during a continuous i.v. injection of secretin (100 U). The S test, however, is an invasive and cumbersome procedure. As a result, N-benzoyl-L-tyrosal-p-aminobenzoic acid (BT-PABA) testing and fecal chymotrypsin testing were introduced into Japan in the middle and late 1970s, respectively. Although simple and noninvasive, these two methods were found have lower sensitivity and specificity than the conventional S test. These two methods, therefore, are presently used more often for monitoring the course of disease and therapeutic effects. Additionally, the glucose tolerance test can be performed to detect endocrine pancreatic insufficiency.

4-Aminobenzoic Acid↗

Prolyl hydroxylase and tissue inhibitor of metalloproteinase in pure pancreatic juice in patients with chronic pancreatitis.

Prolyl hydroxylase is a key enzyme in collagen synthesis, and tissue inhibitor of metalloproteinase (TIMP) is known to suppress collagenolytic enzymes. To see whether the levels of these two enzymes in serum and human pure pancreatic juice (PPJ) are good indicators of pancreatic fibrosis in chronic pancreatitis (CP), we examined 15 controls, 14 alcoholics without evident pancreatic diseases (7 current drinkers and 7 former drinkers), and 19 patients with CP. Levels of the two enzymes were determined by a sandwich enzyme immunoassay method. TIMP-1 levels in PPJ were significantly higher in patients with CP than in controls and alcoholics, with overlap in only a few exceptional patients. A significant inverse correlation between TIMP-1 and bicarbonate output in PPJ was observed. Prolyl hydroxylase levels in PPJ, in contrast, were significantly higher in current drinkers than in patients with CP, controls, and former drinkers, with overlap in only a few exceptional patients with relapsing CP. Identical results were obtained even when the enzyme levels were expressed as nanograms per milligram of protein. Serum levels of prolyl hydroxylase and TIMP-1 showed no significant differences among controls, current alcoholics, former alcoholics, and patients with CP. These results indicate that the raised level of TIMP-1 in PPJ, unlike that of prolyl hydroxylase, is a good indicator of pancreatic fibrosis in CP.

Adult↗

Accumulation of a clock-regulated transcript during flower-inductive darkness in pharbitis nil

To clarify the molecular basis of the photoperiodic induction of flowering in the short-day plant Pharbitis nil cv Violet, we examined changes in the level of mRNA in cotyledons during the flower-inductive photoperiod using the technique of differential display by the polymerase chain reaction. A transcript that accumulated during the inductive dark period was identified and a cDNA corresponding to the transcript, designated PnC401 (P. nil C401), was isolated. RNA-blot hybridization verified that levels of PnC401 mRNA fluctuated with a circadian rhythm, with maxima between 12 and 16 h after the beginning of the dark period) and minima of approximately 0. This oscillation continued even during an extended dark period but was damped under continuous light. Accumulation of PnC401 mRNA was reduced by a brief exposure to red light at the 8th h of the dark period (night-break treatment) or by exposure to far-red light at the end of the light period (end-of-day far-red treatment). These results suggest that fluctuations in levels of PnC401 mRNA are regulated by phytochrome(s) and a circadian clock and that they are associated with photoperiodic events that include induction of flowering.

Journal Article↗

A case of small pancreatic cancer diagnosed by serial follow-up studies promptly by a positive K-ras point mutation in pure pancreatic juice.

We report a 74-yr-old woman who was referred to our hospital because of abdominal fullness. ERP showed a questionable irregularity of the main pancreatic duct at the body. Examination of pure pancreatic juice was positive for K-ras point mutation at codon 12 and negative for cytology. Because neither US nor CT showed apparent lesions in the pancreas, we decided to follow up the patient with serial ERP and pure pancreatic juice studies at 3-month intervals. No changes had been seen up to 18 months later, when cytology was conclusive for malignancy with an apparent stenosis of the main pancreatic duct at the body. Distal pancreatectomy with splenectomy was performed. A round mass, 12 mm in diameter, was found in the body, which proved to be an adenocarcinoma at histological examination. No extrapancreatic extension and metastases were noted. Although positive K-ras point mutation has been reported in some cases of adenoma or mucinous cell hyperplasia of the pancreas and chronic pancreatitis, our case, along with previous reports, indicated the importance of testing K-ras point mutation in pure pancreatic juices for the diagnosis of pancreatic cancer at an early stage.

Adenocarcinoma↗

Establishment of ameloblastoma cell line, AM-1.

Ameloblastomas are slowly growing, locally invasive neoplasms with a potentially destructive behaviour. The molecular mechanisms that regulate the cell growth and invasion of ameloblastoma cells are unknown. Because ameloblastoma cells placed in culture have a very limited lifespan, the establishment of immortalized clones of ameloblastoma cells would aid its study. We produced an immortalized ameloblastoma cell line (AM-1) using human papillomavirus type-16. This cell line maintains epithelial cell morphology and expresses cytokeratins K8, K14, K18, K19. Furthermore, bcl-2 protein, which prevents apoptosis, is expressed. We investigated the behaviour of these cells on a collagen matrix in vitro. These cells grew in a monolayer over foci of collagen degradation and could invade the collagen gel at such sites. Since the behavior of cell line AM-1 mimics the behavior of ameloblastoma in vivo, it may be a valuable model for the study of these neoplasms.

Ameloblastoma↗

Overexpression of bcl-2 protein inhibits terminal differentiation of oral keratinocytes in vitro.

The bcl-2 proto-oncogene is a known inhibitor of apoptosis; in normal human stratified squamous epithelium, its expression is restricted to the basal cell layer. To investigate the functional role of bcl-2 protein in the process of differentiation of oral keratinocytes, bcl-2 expression vector was transfected into SCC-25 cells, which normally undergo squamous cell differentiation in vitro while expressing specific differentiation markers, e.g., keratin 10/11 and involucrin. In bcl-2 transfected SCC-25 cells, the expression of these differentiation markers was markedly suppressed. The bcl-2 proto-oncogene may play a critical role in opposing the commitment to terminal differentiation and apoptosis of oral keratinocytes.

Apoptosis↗

S-1153 inhibits replication of known drug-resistant strains of human immunodeficiency virus type 1.

S-1153 is a new imidazole compound that inhibits human immunodeficiency virus (HIV) type 1 (HIV-1) replication by acting as a nonnucleoside reverse transcriptase inhibitor (NNRTI). This compound inhibits replication of HIV-1 strains that are resistant to nucleoside and nonnucleoside reverse transcriptase inhibitors. S-1153 has a 50% effective concentration in the range of 0.3 to 7 ng/ml for strains with single amino acid substitutions that cause NNRTI resistance, including the Y181C mutant, and also has potent activity against clinical isolates. The emergence of S-1153-resistant variants is slower than that for nevirapine, and S-1153-resistant variants contained at least two amino acid substitutions, including F227L or L234I. S-1153-resistant variants are still sensitive to the nucleoside reverse transcriptase inhibitors zidovudine (AZT) and lamivudine. In a mouse and MT-4 (human T-cell line) in vivo HIV replication model, S-1153 and AZT administered orally showed a marked synergy for the inhibition of HIV-1 replication. S-1153 shows a significant accumulation in lymph nodes, where most HIV-1 infection is thought to occur. S-1153 may be an appropriate candidate for two-to three-drug combination therapy for HIV infection.

Animals↗

Relationship between doubling time of liver metastases from colorectal carcinoma and residual primary cancer.

BACKGROUND: The doubling times of liver metastases were calculated in order to clarify their usefulness in predicting the presence of residual cancer in the abdominal cavity in patients who had undergone curative resection of primary colorectal cancer. PATIENTS AND METHODS: Tumor doubling times were calculated retrospectively in 22 patients by serial measurement of the size of their liver metastases. RESULTS: Patients with a tumor doubling time of less than 92.4 days had a significantly poorer prognosis than those with a doubling time more than or equal to 92.4 days (p < 0.05). Local recurrence or peritoneal dissemination was significantly more likely to occur when the tumor doubling time was less than 92.4 days than when it was more than or equal to 92.4 days (p < 0.01). CONCLUSION: The doubling time of hepatic metastases in patients with colorectal carcinoma may be a useful prognostic indicator, with patients who have a shorter tumor doubling time carrying a greater risk of residual primary cancer in the abdominal cavity.

Cell Division↗

Tissue-specific high-level expression of human endogenous retrovirus-R in the human adrenal cortex.

In an attempt to clarify the biological nature of a human endogenous retrovirus (HERV), HERV-R, which is a single-copy type of HERVs and is conserved as a full-length viral sequence, the expression of HERV-R mRNA in normal autopsied systemic organs was examined by Northern blot analysis. The expression showed different levels among individuals, with the adrenal glands expressing the highest level of HERV-R among all organs tested, except for the placenta. In various adrenal tumors, HERV-R was expressed at high levels in all cortical adenomas but less so in pheochromocytomas. In situ hybridization revealed the expression of HERV-R to be localized in all layers of the adrenal cortex, but not in the medulla. This high-level expression of HERV-R in the adrenal cortex may possibly relate to differentiation and/or steroid production by adrenocortical cells.

Adrenal Cortex↗

Upregulation of retinal vascular endothelial growth factor mRNAs in spontaneously diabetic rats without ophthalmoscopic retinopathy. A possible participation of advanced glycation end products in the development of the early phase of diabetic retinopathy.

Vascular endothelial growth factor (VEGF) has recently been shown to be involved in the pathogenesis of proliferative diabetic retinopathy. However, its involvement in the development of the early phase of diabetic retinopathy is not fully understood. In this study we investigated the retinal VEGF mRNA level in spontaneously diabetic Otsuka Long-Evans Tokushima fatty (OLETF) rats, a model of non-insulin-dependent diabetes, without overt retinopathy, using quantitative reverse-transcription polymerase chain reaction. The retinal VEGF mRNA level was 2.2 times higher (p < 0.0005) in OLETF rats than in control rats at the age of 60 weeks. Moreover, their retinal mRNA level was positively correlated with serum concentration of advanced glycation end products (AGEs) but not to serum glucose concentration. Furthermore, the peak latency of the oscillatory potentials in the electroretinogram, one of the most sensitive markers for the early phase of diabetic retinopathy, was significantly prolonged in OLETF rats (p < 0.05), being also correlated with the serum AGE concentration. The results thus suggest that AGEs, which are formed acceleratedly in diabetic conditions, are involved in the development of the early phase of diabetic retinopathy probably through the induction of retinal VEGF mRNAs.

Animals↗

Identification of Merkel cells by an antibody to villin.

Merkel cells represent a population of epithelial cells in the skin and oral mucosa. Although Merkel cells are reliably distinguishable from other epithelial cells at the ultrastructural level, these cells are usually not discernible by standard light microscopy and need special techniques for their identification. Villin is an actin-crosslinking protein that is associated with the actin filament cores of brush border microvilli. In this study we show that an antibody against villin is an excellent marker of Merkel cells and their microvilli even at the light microscopic level. The surrounding keratinocytes and subepithelial connective tissue cells do not show any significant affinity for the antibody against villin. Confocal laser micrographs reconstructed from serial images 0.5 microm thick of Merkel cells that were immunostained with villin clearly reveal the three-dimensional morphology of Merkel cells and their microvilli. The presence of villin in Merkel cell microvilli lends support to the idea that these cells might have a mechanoreceptor function.

Animals↗

Coherence analysis of EEG changes during olfactory stimulation.

In a pilot study, EEG changes during odor stimulation were evaluated by coherence analysis. Ten normal adults were studied. Simultaneous recordings of 16 EEG channels with and without odor stimulation were stored on magnetic tape for further processing. EEG signals were analyzed using a signal analyzer. Coherence spectra were calculated between all possible channel pairs on the scalp. The amount of data was reduced by extracting broad band coherence values for five frequency bands: delta (2-3.9 Hz), theta (4-7.9 Hz), alpha 1 (8-9.9 Hz), alpha 2 (10-12.9 Hz), and beta 1 (13-17.9 Hz). Coherence values extracted from the control EEG recordings and those during odor stimulation were compared to evaluate the presence of any significant differences. The results demonstrated significant changes in the EEG coherence between the two control recordings (before and after) in the theta and beta 1 bands. These frequency bands were therefore excluded from the examination. During odorant stimulation with methyl-cyclopentenolone, the coherence in the delta band decreased in the frontal region, while that in the alpha 1 and alpha 2 bands increased in the bilateral temporal region. During odorant stimulation with scatol, the coherence in the delta band decreased in the frontal region, while that in the alpha 1 and alpha 2 bands increased in the frontal, temporal and occipital regions. It is suggested that EEG coherence mapping may provide the basis for the development of an objective test of olfactory function in humans.

Adult↗

Cloning of rabbit TR4 and its bone cell-specific activity to suppress estrogen receptor-mediated transactivation.

To clone a new nuclear receptor, we screened a rabbit heart complementary DNA (cDNA) library with degenerate oligonucleotide probes corresponding to the DNA-binding domain of nuclear receptors, which is highly conserved among receptors. One of the cDNA clones, clone 23, encodes a novel protein of 596 amino acids, and predicted molecular mass is 66 kDa. Homology search analysis identified this protein as rabbit TR4 (TR4-0). We also cloned the cDNA encoding a rabbit TR4 isoform (TR4-1), which lacks the putative C-terminal ligand-binding domain (350 amino acids) caused by a 23-bp exon deletion, which probably occurred during messenger RNA (mRNA) splicing. Northern blot analysis showed that TR4s are expressed with two kinds of mRNAs (9.0 kb and 2.8 kb), both of which are relatively abundant in brain, testis, and bone. RT-PCR analysis, using pairs of primers specific for each TR4, showed that both types of receptor express in various tissues. Furthermore, both are present in primary osteoblasts and bone marrow cells, though the mRNA levels of TR4-0 were much higher than those of TR4-1. A functional study, using a transient transfection assay, showed that both receptors suppressed retinoid X receptor (RXR)-retinoid acid receptor, RXR-TR, and RXR-VDR-mediated transactivation significantly in COS-1 and osteosarcoma cells (UMR-106, ROS17/2.8) and that TR4-0 was much more effective than TR4-1. Unexpectedly, we found that the TR4s effectively suppressed estrogen receptor-mediated transactivation in bone cells, but neither in kidney (COS-1) nor breast cancer cells (MCF-7, one of the major target cells of the estrogen action). Thus, the present study shows a novel property of the TR4 orphan receptor, acting as a bone cell-specific repressor in the estrogen receptor-mediated signaling pathway.

Amino Acid Sequence↗

Involvement of lymphocyte function-associated antigen-1 and intercellular adhesion molecule-1 in osteoclastogenesis: a possible role in direct interaction between osteoclast precursors.

In our search for molecules involved in the process of osteoclast differentiation, we examined the surface phenotypes of the preosteoclast-like cells and osteoclast-like multinucleated cells (MNCs) formed in bone marrow cultures, using monoclonal antibodies recognizing different antigen molecules expressed on hematopoietic cells. Among these cell surface antigens, lymphocyte function-associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1) were highly expressed on mononuclear cells in the cultures for forming preosteoclast-like mononuclear cells. The double detection of these two antigen molecules with osteoclast-specific antigen and with calcitonin receptor, using a fluorescence-activated cell sorter or autoradiography technique, revealed that LFA-1 and ICAM-1 were expressed on the preosteoclasts. The expression of ICAM-1 was detected on both preosteoclasts and osteoclast-like MNCs, whereas the expression of LFA-1 was restricted to preosteoclasts. We designed a peptide with the sequence of the binding site of ICAM-1 against the ligand LFA-1. In the whole bone marrow culture system for forming osteoclast-like MNCs, a significant inhibition of MNC formation was observed by the addition of this peptide. These results strongly suggest the involvement of an LFA-1/ICAM-1-interaction in osteoclastogenesis.

Animals↗

Dose-dependent selective cytotoxicity of extracts from marine green alga, Cladophoropsis vaucheriaeformis, against mouse leukemia L1210 cells.

The selective cytotoxic activity of extracts from two marine green algae, Cladophoropsis vaucheriaeformis and Halimeda discoidea, was examined via a dose response assay against mouse leukemia L1210 cells and normal NIH-3T3 cells. The MeOH-extract from C. vaucheriaeformis showed selective cytotoxicity to L1210 cells at concentrations ranging from 50 to 100 microg/ml. In particular, the greatest selectivity for cytotoxic activity was found at the concentration of 50 microg/ml, at which the growth of L1210 cells was inhibited completely and that of NIH-3T3 was not affected at all. However, MeOH extracts from the red alga Laurencia okamurae and the brown alga Dictyopteris undulata, which displayed non-selective cytotoxicity in our previous screening program, did not show similar selective cytotoxicity at any concentrations tested. These results indicate that the marine green alga C. vaucheriaeformis may contain a unique antitumor substance with selective cytotoxic activity against L1210 cells. Our results also suggest that this active substance might be of low molecular weight and therefore MeOH-soluble.

Animals↗

Gastrobiliary dysmotility in patients with chronic pancreatitis as assessed by a single noninvasive test.

We simultaneously assessed gastric emptying and gallbladder contraction after oral administration of a liquid meal by noninvasive ultrasonography in 17 patients with chronic pancreatitis (CP) and in 17 healthy controls. Gastrointestinal (GI) transit was also assessed by a noninvasive radioopaque marker method. Exocrine pancreatic function was evaluated by analyzing pure pancreatic juice and by analyzing the autonomic nervous system by cardiovascular reflex tests. Patients with CP showed impaired gallbladder contraction at 15 min and hastened gastric emptying. The cause of the former is unclear, whereas the latter was closely related with decreased pancreatic lipase output, but not with autonomic dysfunction. GI transit time did not differ between controls and patients with CP. In conclusion, we succeeded in clearly demonstrating impaired gallbladder contraction and hastened gastric emptying in patients with CP by a single noninvasive test, ultrasonography. We also revealed for the first time that hastened gastric emptying is associated with insufficient pancreatic lipase output.

Autonomic Nervous System Diseases↗

Estimation of thermal sensation during varied air temperature conditions.

Seven male students were exposed to four varied air temperature environments: hot (37 degrees C) to neutral (27 degrees C) (HN), neutral to hot (NH), cool (17 degrees C) to neutral (CN), and neutral to cool (NC). The air temperature was maintained at the first condition for 20 min, then was changed to the second condition after 15 min and was held there for 20 min. Each subject wore a T-shirt, briefs, trunks, and socks. Each sat on a chair and was continuously evaluated for thermal sensation, thermal comfort, and air velocity sensation. Some physiological and thermal parameters were also measured every 5 s during the experiment. The correlation between thermal sensation and skin temperature at 15 sites was found to be poor. The subjects felt much warmer during the rising phase of the air temperature (CN, NH) than during the descending phase (HN, NC) at a given mean skin temperature. However, thermal sensation at the same heat flux or at the same value of the difference between skin and air temperature (delta(Tsk - Ta)) was not so different among the four experimental conditions, and the correlation between thermal sensation and heat flux or delta(Tsk - Ta) was fairly good. The multiple regression equation of the thermal sensation (TS) on 15 sites of skin temperature (Tsk; degrees C) was calculated and the coefficient of determination (R*2) was found to be 0.656. Higher coefficients of determination were found in the equations of thermal sensation for the heat flux (H; kcal.m-2.h-1) at the right and left thighs of the subjects and on delta(Tsk - Ta) (degrees C) at 4 sites. They were as follows: TS = 2.04 - 0.016 Hright - 0.036 Hleft; R*2 = 0.717, TS = 1.649 + 0.013 delta(Tsk - Ta)UpperArm - 0.036 delta(Tsk - Ta)Chest - 0.223 delta(Tsk - Ta)Thigh-0.083 delta(Tsk - Ta)LowerLeg; R*2 = 0.752, respectively.

Adult↗

Safety management of a patient undergoing thoracic aortic surgery by spinal evoked potential monitoring.

We have presented a patient (48 year-old male) in whom the evoked spinal potential monitor detected impending spinal ischemia after aortic cross-clamping, which allowed surgical intervention to be modified so as to restore the decaying evoked spinal potential. The patient received replacement of the dissecting aneurysm in the thoracic descending aorta with clamping of the aorta at the sites immediately proximal and distal to the aneurysm and initiation of femoro-femoral venoarterial bypass under normothermia. The evoked spinal potential was recorded via the T12/L1 epidural electrode in response to transdural electric stimulation of the spinal cord at the C7/T1 level. As the evoked spinal potential gradually decreased in amplitude without latency prolongation after aortic cross-clamping, the distal clamp was moved from the T6 vertebral level to the T4. The reduced spinal potential then returned to the baseline amplitude. This episode was repeated twice from surgical necessity. The patient was free from any neurological disorders postoperatively.

Aortic Dissection↗