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Biomedical subjects

T Aoyagi

Publications and source records attributed to T Aoyagi.

At least 73 records · Page 4Linked to original sources
PubMed1999

Sildenafil induced priapism.

A 53 year-old Japanese man was referred to our hospital for persistent priapism, which had been induced by 200 mg (usual dose 25-50 mg) of sildenafil citrate (Viagra) three days earlier. He had a history of erectile dysfunction and had undergone penile injection therapy elsewhere; however, he had not used injection therapy this time. He obtained sildenafil personally without a doctor's prescription. He had not taken any other drugs that affect the metabolism of sildenafil, nor did he have any medical complications that might induce priapism. Since needle aspiration and irrigation were ineffective as first line therapy, surgical treatment was indicated to relieve the condition; a incision of tunica albuginea of both corpora cavernosa was made, and vigorous irrigation of saline washed out the blood clots. This is the first case report of priapism induced by sildenafil. Although this drug can be obtained through private commerce, it should be used under professional guidance as its abuse may lead to severe morbidity.

Humans↗
PubMed1999

The absolute configuration of belactin A, a beta-lactone-containing serine carboxypeptidase inhibitor: importance of the beta-lactone structure for serine carboxypeptidase inhibition.

The absolute configuration of belactin A, a beta-lactone-containing serine carboxypeptidase inhibitor was studied by a crystal X-ray diffraction analysis and its absolute structure was determined to be (2R,3S)-2-¿(3S)-3-[(2-amino-5-chlorophenyl)carboxamido]-1,1-dimethyl-2-o xobutyl¿-3-methyl-4-oxooxetane. The importance of the beta-lactone structure for inhibitory activity was found by preparing several derivatives of belactin A.

Benzamides↗
PubMed1999

[The use of predeposited autologous blood transfusion for radical prostatectomy and total cystectomy].

BACKGROUND: To avoid homologus blood transfusion, we performed the operation for intra-pelvic malignancy with predeposited autologus blood using recombinant human erythropoietin (rH-EPO). MATERIALS AND METHODS: The seven cases of radical prostatectomy and the 15 cases of total cystectomy were analyzed retrospectively. All cases were given ferrons sulfate/200 mg of iron orally every day to the day before the operation and treated with 24,000 unit of rH-EPO subcutaneously every week. The target volumes of preoperative autologus blood collection were 800 to 1,000 ml and 800 to 1,200 ml for radical prostatectomy and total cystectomy, respectively. For each case, 400 ml blood was collected once a week. RESULTS: In cases of radical prostatectomy, the preserved blood volume was 885.7 +/- 157.4 ml and 6 out of 7 operations were successfully performed without additional homologus blood transfusion (85.7%). In total cystectomy, the preserved blood volume was 1,033.3 +/- 167.6 ml and 14 out of 15 operations were successfully performed without additional homologus blood transfusion (93.3%). The bleeding volume during operation showed no significant difference compared to control group where used homologus blood transfusion. Postoperative courses were uneventful and there encountered no severe side effects and complications in all our procedures. CONCLUSIONS: Our study indicates that in cases of radical prostatectomy and total cystectomy, the operation with predeposited autologus blood using rH-EPO is possible to be performed in safe. The effect on long term prognosis of malignancy is not clear, however, this technique is helpful able to avoid hazardous issues related to homologus blood transfusion during the operation.

Aged↗
PubMed1999

Insulin action on heart and skeletal muscle FDG uptake in patients with hypertriglyceridemia.

UNLABELLED: Abnormal heart and skeletal muscle 18F-fluorodeoxyglucose (FDG) uptake in patients with insulin resistance has been demonstrated. Although the existence of whole-body insulin resistance has been reported in hypertriglyceridemics, its specific role in heart and skeletal muscle FDG uptake in hypertriglyceridemics has not been clarified. METHODS: We compared heart and skeletal muscle FDG uptake using PET and the whole-body glucose disposal rate (GDR) during insulin clamping in 17 hypertriglyceridemics and 12 age-matched control subjects to increase our knowledge of whole-body insulin resistance and its relationship to heart and skeletal muscle FDG uptake in hypertriglyceridemics. RESULTS: GDR was significantly reduced in hypertriglyceridemics compared with control subjects (4.50 +/- 1.37 mg/min/kg versus 10.0 +/- 2.97 mg/min/kg, P = 0.00001), as were the skeletal muscle FDG Ki = (k1 x k3)/(k2 + k3) (SFKi: 0.007 +/- 0.003 mL/min/g versus 0.018 +/- 0.01 mL/min/g, P = 0.0001) and skeletal muscle FDG uptake ([SMFU] 0.725 +/- 0.282 mg/min/100 g versus 1.86 +/- 1.06 mg/min/100 g, P = 0.00023). However, myocardial FDG Ki (MFKi) tended to be reduced in hypertriglyceridemics compared with that in control subjects (0.062 +/- 0.017 mL/min/g versus 0.068 +/- 0.015 mL/min/g), but the difference was statistically insignificant (P = 0.3532). Moreover, myocardial FDG uptake (MFU) in hypertriglyceridemics (6.47 +/- 1.72 mg/min/100 g) tended to be reduced compared with that in control subjects (6.97 +/- 1.73 mg/min/100 g), but the difference was statistically insignificant (P = 0.4485). GDR was significantly correlated with SFKi (r = 0.69, P = 0.0022), SMFU (r = 0.612, P = 0.009), MFKi (r = 0.57, P = 0.0174) and MFU (r = 0.505, P = 0.0385) in hypertriglyceridemics. CONCLUSION: Both heart and skeletal muscle glucose utilization were related to insulin resistance in hypertriglyceridemics. However, the less severe reduction in MFU (compared with SMFU) suggests that myocardium may have a mechanism to oppose insulin resistance in hypertriglyceridemics.

Case-Control Studies↗
PubMed1999

[Comparison of the newly developed MB redox system with mycobacteria growth indicator tube (MGIT) and 2% Ogawa egg media for recovery of mycobacteria in clinical specimens].

The rate of recovery and the mean time to detection of mycobacteria in clinical specimens were determined in a newly-developed MB Redox system based on liquid medium, and the results were compared with those of MGIT and 2% Ogawa egg media. From 587 sputum specimens processed, totally 203 mycobacterial isolates were detected, of which 177 (87.2%) with MB Redox, 185 (91.1%) with MGIT and 133 (65.6%) with 2% Ogawa medium. The difference in the percentages of positive cultures between either of the two liquid media and 2% Ogawa medium was significant (p < 0.0001). The mean time to detection of the Mycobacterium tuberculosis complex was 17.5 days with MB Redox, 18.7 days with MGIT, and 26.2 days with 2% Ogawa medium. The contamination rates were 1.5, 1.7, and 4.1% for MB Redox, MGIT, and 2% Ogawa medium, respectively. In conclusion, both MB Redox and MGIT systems, based on liquid medium, are more efficient than 2% Ogawa medium for the recovery of mycobacteria in clinical specimens.

Bacteriological Techniques↗
PubMed1998

Preparation and characterization of thermally responsive block copolymer micelles comprising poly(N-isopropylacrylamide-b-DL-lactide).

The thermally sensitive block copolymer, poly(N-isopropylacrylamide-b-dl-lactide) (PIPAAm-PLA), was synthesized by ring-opening polymerization of dl-lactide initiated from hydroxy-terminated poly (N-isopropylacrylamide) (PIPAAm). A PIPAAm bearing a single terminal hydroxyl group was prepared by telomerization using 2-hydroxyethanethiol as a chain-transfer agent. Successful preparation of PIPAAm and the PIPAAm-PLA block copolymer was verified by gel permeation chromatography (GPC) and 1H-NMR spectroscopy. Polymeric micelles were prepared from block copolymers using a dialysis method. Their solutions showed reversible changes in optical properties: transparent below a lower critical solution temperature (LCST) and opaque above the LCST. Dynamic light scattering measurements were used to observe the formation of micellar structures approximately 40 nm in diameter, which do not change between 20 degreesC and 30 degreesC. Above the LCST, polymer micelles aggregated, a phenomenon found to be reversible since the aggregates dissociated again by cooling below the LCST. Further observations using atomic force microscopy (AFM) confirmed this behaviour. The properties of this block copolymer system are interesting from both applied and fundamental perspectives, particularly for active targeting as drug carriers.

Acrylamides↗
PubMed1998

Retrieval dynamics in oscillator neural networks.

We present an analytical approach that allows us to treat the long-time behavior of the recalling process in an oscillator neural network. It is well known that in coupled oscillatory neuronal systems, under suitable conditions, the original dynamics can be reduced to a simpler phase dynamics. In this description, the phases of the oscillators can be regarded as the timings of the neuronal spikes. To attempt an analytical treatment of the recalling dynamics of such a system, we study a simplified model in which we discretize time and assume a synchronous updating rule. The theoretical results show that the retrieval dynamics is described by recursion equations for some macroscopic parameters, such as an overlap with the retrieval pattern. We then treat the noise components in the local field, which arise from the learning of the unretrieved patterns, as gaussian variables. However, we take account of the temporal correlation between these noise components at different times. In particular, we find that this correlation is essential for correctly predicting the behavior of the retrieval process in the case of autoassociative memory. From the derived equations, the maximal storage capacity and the basin of attraction are calculated and graphically displayed. We also consider the more general case that the network retrieves an ordered sequence of phase patterns. In both cases, the basin of attraction remains sufficiently wide to recall the memorized pattern from a noisy one, even near saturation. The validity of these theoretical results is supported by numerical simulations. We believe that this model serves as a convenient starting point for the theoretical study of retrieval dynamics in general oscillatory systems.

Artifacts↗
PubMed1998

Depletion of arylhydrocarbon receptor during adipose differentiation in 3T3-L1 cells.

Arylhydrocarbon receptor (AhR) is the receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds. Although a physiological ligand for AhR has yet to be identified, several lines of evidence suggest that AhR may play an important role not only in the regulation of xenobiotic metabolism but also in the maintenance of homeostatic functions. When TCDD is administrated in vivo, this compound is primarily deposited in adipose tissue. Therefore, it is critical to know the states of AhR in adipose cells for assessing the expression of toxicities of TCDD and related compounds in vivo. In this report, we examined the levels of AhR protein and its associated protein (Arnt) during the adipose differentiation in 3T3-L1 cells. The level of AhR protein was found to decrease with ongoing adipose differentiation in 3T3-L1 cells. The binding activity to the xenobiotic response element and the cellular response to TCDD were also lowered as a result of adipose differentiation. These results indicate that the depletion of AhR is a novel event associated with adipose differentiation in 3T3-L1 cells and that the magnitude of the depletion of AhR is sufficient for 3T3-L1 cells to lose the functional response to xenobiotics. We also found a population of 3T3-L1 cells which have an adipose differentiation capability in the presence of high doses of TCDD. These cells lack nuclear AhR but not cytoplasmic AhR, suggesting a possible negative role of liganded nuclear AhR in adipose differentiation. The level of the Arnt protein also decreased as a result of the differentiation. However, the pattern of the depletion of the Arnt protein was distinct from that of the AhR protein. The data presented in this study will provide opportunities to carry out studies to better understand the roles of AhR in adipose cells which are the primary targets of TCDD.

3T3 Cells↗
PubMed1998

Insulin-like growth factor I stimulates proliferation and Fas-mediated apoptosis of human osteoblasts.

In vitro studies have shown that insulin-like growth factor I (IGF-I) is a potent growth factor for osteoblasts, although both bone formation and resorption are upregulated by IGF-I in vivo. To understand the difference in the action of IGF-I observed in vitro and in vivo experiments, we examined the effect of IGF-I on the proliferation and Fas-mediated apoptosis of human osteoblasts in vitro. Human osteoblastic cell line MG63 and human primary osteoblast-like cells obtained from biopsy specimens were used as human osteoblasts. Cells were cultured with or without various concentrations of IGF-I followed by determination of the proliferative response and Fas-mediated apoptosis. IGF-I dose dependently stimulated the proliferation of cultured human osteoblasts. Both Fas expression and the degree of anti-Fas IgM-induced apoptosis of human osteoblasts was also augmented by IGF-I. Furthermore, the cytotoxicity of Fas ligand (FasL) cDNA transformants against human osteoblasts was increased when IGF-I-stimulated osteoblasts were used as target cells, indicating that stimulation of IGF-I increased functional Fas expression on human osteoblasts as well as their proliferation. The addition of DEVD-CHO, a specific inhibitor of CPP32, to the culture resulted in a significant inhibition of Fas-mediated apoptosis of both unstimulated and IGF-I-stimulated osteoblasts, although it did not affect the proliferative response or Fas expression. Our data suggest that activation of CPP32 is necessary for Fas-mediated apoptosis of human osteoblasts, and treatment of IGF-I increased this signaling pathway. In contrast, regulation of proliferation and Fas expression of the cells were probably not affected by CPP32 activation. Our results suggest that IGF-I acts on cultured human osteoblasts by increasing their proliferation and induction of Fas-mediated apoptosis by neighbouring FasL+ cells such as osteoclasts, thus probably functioning as a local coupling factor in the bone in vivo, stimulating both bone formation and resorption.

Apoptosis↗
PubMed1998

Effect of molecular architecture of hydrophobically modified poly(N-isopropylacrylamide) on the formation of thermoresponsive core-shell micellar drug carriers.

Terminal-incorporation of hydrophilic or hydrophobic groups dramatically influences the phase transition of poly(N-isopropylacrylamide) (PIPAAm) because of a critical role of the polymer chain ends in initiation of the phase transition. Incorporation of an amino or hydroxyl group to one end of PIPAAm remarkably raised the LCST (lower critical solution temperature) and slowed down the rate of the phase transition, and these effects were more pronounced as the mole fraction of hydrophilic groups increased compared to the random copolymers of PIPAAm and hydrophilic co-monomers, such as acrylic acid (AAc) or dimethylacrylamide (DMAAm). Hydrophilic effects were more remarkable for hydroxyl groups, due to stronger hydrogen bonding with water. Terminal-modification (hydrophobization) was also more effective in producing hydrophobic effects on the PIPAAm phase transition in comparison with PIPAAm copolymers that were randomly modified along the main chain with hydrophobic co-monomers. Moreover, terminal-located hydrophobic groups were able to form hydrophobic microdomains that were clearly isolated from PIPAAm chains in aqueous media by the aggregation of hydrophobic segments. As a result, the obtained micellar aqueous solution showed the same LCST as pure PIPAAm, while the PIPAAm random copolymer with hydrophobic co-monomers formed incompletely separated microdomains. The LCST for this random copolymer was reduced with increasing hydrophobic co-monomer mole fraction. Hydrophobically terminal-modified PIPAAm produced thermo-responsive core-shell structures that exhibited the same LCST and the same thermal response rate as those of free linear PIPAAm chains. Such polymeric micellar structures show reversible thermoresponsive aggregation/dispersion and deformation/reformation in heating/cooling cycles through the LCST for pure PIPAAm. These properties indicate the possibility of using such a system as a thermoresponsive drug carrier with double targeting mechanisms, in both passive and active manners.

Acrylic Resins↗
PubMed1998

Role of positron emission tomography using fluorine-18 fluoro-2-deoxyglucose in predicting improvement in left ventricular function in patients with idiopathic dilated cardiomyopathy.

Improvement in left ventricular (LV) function in patients with idiopathic dilated cardiomyopathy (DCM) by medical treatment has been suggested. Thus, it is important to evaluate which patients will respond to medical therapy. Positron emission tomography (PET) with fluorine-18 fluoro-2-deoxyglucose (FDG) and cardiac catheterization were performed in 20 patients with DCM before the initiation of medical therapy. The regional myocardial glucose utilization rate (rMGU) was measured with FDG PET. Subjects were divided into two groups, group 1 (event-free patients, n=10) and group 2 (clinical cardiac events, n=10). Haemodynamic and PET parameters before the initiation of medication were compared between the two groups and between patients with and patients without improvement in LV function. Ejection fraction (EF) was significantly higher in group 1 (35.8%+/-9.0%) than in group 2 (24.8%+/-7.0%) and LV end-diastolic pressure (LVEDP) was significantly lower in group 1 (8.4+/-1.7 mmHg) than in group 2 (11.6+/-3.5 mmHg). Average rMGU (mg min-1 100 g-1) was similar in group 1 (11.2+/-2.5 mg min-1 100 g-1) and group 2 (11.2+/-2.9 mg min-1 100 g-1), while %CV of rMGU was significantly lower in group 1 (11.1%+/-6.3%) than in group 2 (29. 9%+/-13.9%, P<0.01). Furthermore, LV function normalized in seven patients in group 1. In these seven patients, EF (35.1%+/-10.9%), LVEDP (8.2+/-2.0 mmHg) and average rMGU (11.8+/-2.7 mg min-1 100 g-1) were comparable with those in patients without LV functional improvement (EF: 31.6%+/-9.1%; LVEDP: 10.7+/-3.3 mmHg; average rMGU: 10.8+/- 2.7 mg min-1 100 g-1). However,% CV of rMGU in patients with LV functional improvement (9.6%+/-5.6%) was significantly lower than in those without such improvement (26.3%+/-14.1%, P<0.01). %CV of rMGU <13.6% predicted prognosis with a sensitivity of 80%, a specificity of 100% and an accuracy of 90%. %CV of rMGU <13.6% also predicted improvement in LV function, with a sensitivity of 75%, a specificity of 92% and an accuracy of 85%. However, EF failed to predict improvement of LV function. In is concluded that homogeneous myocardial glucose utilization rate can predict both prognosis and improvement in LV function achieved by medical therapy in patients with DCM.

Adult↗
PubMed1998

Apoptosis induction in synovial fibroblasts by ceramide: in vitro and in vivo effects.

Several lipid second messengers are important mediators of extracellular signals. Among them ceramide, which is formed by cell membrane sphingomyelin, influences the apoptotic signal pathway through Fas antigen. We examined the apoptotic effect of cell-permeable C2-ceramide on rheumatoid synovial fibroblasts in vitro and in vivo. Exposure of cultured human rheumatoid synovial fibroblasts to C2-ceramide for 24 hours produced internucleosomal DNA fragmentation and morphologic changes characteristic of apoptosis. This C2-ceramide-mediated apoptosis was dose dependent as confirmed by analysis of cytosolic oligonucleosome-bound DNA of treated synovial fibroblasts. We also demonstrated that intra-articular administration of C2-ceramide into Fas-deficient MRL lpr/lpr mice produced a profound reduction of synovial hyperplasia within 24 hours. In situ nick end labeling analysis confirmed the induction of apoptosis in synovial lining cells. Our results indicate that C2-ceramide may function as a potent inducer of apoptosis in the synovium and suggest that pharmacologically-induced apoptosis may be useful as a new therapeutic modality for rheumatoid arthritis.

Animals↗
PubMed1998

Visualization of PEO-PBLA-pyrene polymeric micelles by atomic force microscopy.

PURPOSE: To directly visualize and evaluate the aqueous block copolymeric micelles, poly(ethylene oxide)-poly(beta-benzyl L-aspartate) (PEO-PBLA) chemically conjugated with pyrene fluorescence molecule, by nanotechnology of atomic force microscopy (AFM). METHODS: The block copolymers' PEO-PBLA-Pyrene was first synthesized by reacting with pyrene sulfonyl chloride and PEO-PBLA in tetrahydrofuran (THF) solution and were identified by GPC reflect index, UV and fluorescence detectors. The characterization of physical and chemical properties of PEO-PBLA-Pyrene polymeric micellar solution were examined by the dynamic light scattering (DLS) and critical micelles concentrations (CMC). In addition, the nanotechnology of AFM was used to directly visualize the size and shape of nanopolymeric micelles. RESULTS: The pyrene fluorescence molecule were successfully conjugated at the amino group of the end of PBLA chain by GPC with three different detectors. The size of the aqueous PEO-PBLA-Pyrene polymeric micelles was detected around 57 nm with unimodal distribution by DLS measurement. As a result of this finding, the CMC test was also found out that the fluorescence intensity was increasing around 0.01 approximately 0.05 mg/ml. Using AFM evaluation of polymeric micellar solution, the morphology of aqueous PEO-PBLA-Pyrene polymeric micelles was observed on round shape and with the narrow dispersity of size range 50 approximately 80 nm. CONCLUSIONS: The presence of PEO-PBLA copolymers with pyrene in an aqueous system formed in a spherical and nano range of polymeric micelles.

Micelles↗
PubMed1998

CD4+ T-cell-mediated cytotoxicity against staphylococcal enterotoxin B-pulsed synovial cells.

Apoptosis of synovial cells in rheumatoid arthritis (RA) synovium determined in vivo is suggested to counteract the overgrowth of synovium. Immunohistological examination has revealed the infiltration of activated CD4+ T cells, which express Fas ligand (FasL), in RA synovium. The presence of a putative antigen (Ag) of autoimmune disorders in a target organ may induce the activation of specific T cells in the inflammatory region such as RA synovium. We examined the possible role of CD4+ T cells activated by synovial cells in a staphylococcal enterotoxin B (SEB)-dependent manner, inducing synovial cell apoptosis. Synovial cells were cultured with or without interferon-gamma (IFN-gamma) and further incubated with CD4+ T cells in the presence of SEB. After the cocultivation, both the cytotoxicity and FasL expression of CD4+ T cells were investigated. Constitutive Fas expression was detected on both unstimulated and IFN-gamma-stimulated synovial cells. CD4+ T cells did not kill SEB-pulsed unstimulated synovial cells efficiently. In contrast, when CD4+ T cells were incubated with IFN-gamma-stimulated synovial cells with SEB whose human leucocyte antigen (HLA)-DR and -DQ expression was markedly induced, significant cytotoxicity by these cells against synovial cells was detected. The addition of anti-HLA-DR and -DQ monoclonal antibodies (mAbs) or human Fas chimeric protein (hFas-Fc) reduced this cytotoxicity. FasL expression of CD4+ T cells cocultured with IFN-gamma-stimulated synovial cells with SEB was significantly induced. Furthermore, the addition of mAbs against CD54, CD58 and CD106 inhibited both the cytotoxicity and FasL expression of CD4+ T cells induced by IFN-gamma-stimulated synovial cells in the presence of SEB, indicating the importance of costimulatory molecules on synovial cells in activating CD4+ T cells. Our results suggest that CD4+ T cells are activated by synovial cells by an SEB-dependent manner and express FasL, inducing Fas-mediated apoptosis of the latter cells. These phenomena may regulate the overgrowth of synovial cells in RA synovium.

Antibodies, Monoclonal↗
PubMed1998

Blood volume measurement at the bedside using ICG pulse spectrophotometry.

BACKGROUND: In the treatment of critically ill patients, blood volume (BV) measurement requires injection of some tracer substance and subsequent blood sampling to analyze the tracer concentration. To obviate both the sampling and laboratory analysis, techniques of pulse oximetry have been adapted to the noninvasive optical measurement in the patient's nose or finger of the arterial concentration of an injectable dye. METHODS: The authors report the clinical accuracy of a new noninvasive bedside BV measurement test that uses pulse spectrophotometry (the pulse method). The device detects pulsatile changes of tissue optical density of a nostril or a finger spanned by a probe emitting two infrared wavelengths (805 and 890 nm). After a peripheral or central intravenous injection of indocyanine green, the arterial dye concentration is continuously computed by reference to the previously measured blood hemoglobin concentration. Three types of tests of its accuracy are described here. RESULTS: In 10 healthy volunteers, the authors compared BV determined by the pulse method with an (131)I-labeled human serum albumin method. Three subject data sets were excluded because of motion artifact, a low signal:noise ratio, or both. For the other seven volunteers, the bias+/-SD of pulse spectrophotometric BV values were 0.20+/-0.24 l (or 4.2+/-4.9%) for the nose probe and 0.34+/-0.31 l (or 7.3+/-6.9%) for the finger probe, with a mean BV of 5 l. In 30 patients who underwent cardiac surgery, the pulse method was compared with a standard indocyanine green method using intermittent blood sampling. In three patients, the BV could not be determined by the pulse method because of motion artifact, low signal:noise ratio, or both. In 27 patients, the bias+/-SD of the BV by the pulse method was -0.23+/-0.37 l (-5.3+/-8.7%) for the nose and -0.25+/-0.5 l (-4.2+/-8.4%) for the finger. Patient BV ranged from 2.51 to 7.13 l (mean, 4.48 l). In 10 additional patients before cardiac surgery, BV was measured by the pulse method before and shortly after removal of 400 ml blood. The pulse method recorded a decrease of BV of 480+/-114 mL Three days after venesection, the mean BV was 117+/-159 ml less than the predonation control. CONCLUSIONS: In most patients, the pulse method provides bedside measurement of BV without blood sampling (except for hemoglobin determination), with an estimated error less than 10%. In 10-30% of tests the method failed because of motion distortion of the record during the 10-min data collection period or because of insufficient pulse amplitude in the test tissue.

Adult↗