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Biomedical subjects

H Yoshida

Publications and source records attributed to H Yoshida.

At least 433 records · Page 24Linked to original sources
PubMed2000

Retrovirus-mediated gene transfer of granulocyte colony-stimulating factor receptor (G-CSFR) cDNA into MDS cells and induction of their differentiation by G-CSF.

Myelodysplastic syndromes (MDS) are clonal disorders in which the proper differentiation of hematopoietic stem cells is impaired. There is no effective treatment for this stem cell disorder at present. In an attempt to find a new strategy that promotes the differentiation of MDS blast cells, we tried retroviral transduction of granulocyte colony-stimulating factor receptor (G-CSFR) into an interleukin-3-dependent MDS cell line, MDS-L, since expression of G-CSFR is known to be essential for the differentiation of myeloid progenitor cells and this expression is impaired in most MDS cells. Ectopic expression of human G-CSFR cDNA in MDS-L cells gave rise to granulocytic differentiation by G-CSF stimulation. G-CSF caused the transformants expressing G-CSFR to display a morphological characteristic of mature granulocytes, upregulated CD11b on the cell surface, and improved NBT reduction activity. These results demonstrate that MDS-L cells ecopically expressing G-CSFR are induced to granulocytic differentiation upon exposure to G-CSF, and shed light on the molecular mechanisms of maturation arrest in MDS cells.

Antigens, CD↗
PubMed2000

The effects of monotherapy or combined therapy with an angiotensin converting enzyme inhibitor following initial treatment with calcium channel blockers on residual cardiovascular abnormalities.

The effects of sequential therapy with angiotensin-converting enzyme inhibitor (cilazapril) on left ventricular (LV) mass, LV diastolic function, and carotid artery distensibility were evaluated in 90 hypertensive patients whose blood pressure controlled below 140/90 mmHg with a calcium channel blocker monotherapy. The possibility of predicting the efficacy of cilazapril based on evaluation of biochemical and genetic markers of the renin-angiotensin system was examined. Before cilazapril therapy, LV diastolic function and carotid artery distensibility were significantly impaired in 32 patients with residual LV hypertrophy compared with patients without LV hypertrophy. Cilazapril improved the LV mass in these patients with LV hypertrophy and improved LV diastolic function in a subset of 20 patients with elevated plasma renin activity. Patients with residual LV hypertrophy accompanied by cardiovascular functional abnormalities. Subsequent treatment with cilazapril significantly improved LV morphology and function in those with residual LV hypertrophy or elevated plasma renin activity.

Aged↗
PubMed2000

Involvement of a small GTP-binding protein (G protein) regulator, small G protein GDP dissociation stimulator, in antiapoptotic cell survival signaling.

Small GTP-binding protein GDP dissociation stimulator (Smg GDS) regulates GDP/GTP exchange reaction of Ki-Ras and the Rho and Rap1 family members and inhibits their binding to membranes. In fibroblasts, Smg GDS shows mitogenic and transforming activities in cooperation with Ki-Ras. However, the physiological function of Smg GDS remains unknown. Here we show that mice lacking Smg GDS died of heart failure shortly after birth, not resulting from developmental heart defects but from enhanced apoptosis of cardiomyocytes triggered by cardiovascular overload. Furthermore, neonatal thymocytes and developing neuronal cells underwent apoptotic cell death. Smg GDS-/- thymocytes were susceptible to apoptotic inducers, such as etoposide and UV irradiation. Smg GDS-/- thymocytes were protected from etoposide-induced cell death by ex vivo transduction of the Smg GDS cDNA. These phenotypes partly coincide with those observed in Ki-Ras-deficient mice, suggesting that Smg GDS is involved in antiapoptotic cell survival signaling through Ki-Ras.

Animals↗
PubMed2000

Walking speed as a good predictor for the onset of functional dependence in a Japanese rural community population.

OBJECTIVE: to investigate and compare the predictive values of four physical performance measures for the onset of functional dependence in older Japanese people living at home. DESIGN: a population-based prospective cohort study. SETTING: Nangai village, Akita Prefecture, Japan. METHODS: out of the population aged 65 years and older living in Nangai (n = 940) in 1992, we measured hand grip-strength, one-leg standing, and usual and maximum walking speeds in 736 subjects who were independent in the five basic activities of daily living. Their functional status was assessed each year for the subsequent 6 years. The outcome event was the onset of functional dependence, defined as a new disability in one or more of the five basic activities of daily living, or death of a subject who had shown no disability at the previous follow-up. RESULTS: even after controlling for age, sex and a number of chronic conditions, lower scores on each baseline performance measure showed increased risk for the onset of functional dependence. Maximum walking speed was most sensitive in predicting future dependence for those aged 65-74 years, while usual walking speed was most sensitive for people aged > or = 75 years. CONCLUSION: walking speed was the best physical performance measure for predicting the onset of functional dependence in a Japanese rural older population.

Activities of Daily Living↗
PubMed2000

Induction of murine intestinal and hepatic peroxiredoxin MSP23 by dietary butylated hydroxyanisole.

Feeding mice with 2(3)-t-butyl-4-hydroxyanisole (BHA) induces phase II detoxifying enzymes that inhibit the action of carcinogens. We have found that dietary BHA induces intestinal and hepatic MSP23 (also called peroxiredoxin I), a stress-inducible antioxidant, in a manner similar to the induction of glutathione S-transferases (GSTs). The levels of MSP23 in the proximal intestine and liver, estimated by immunoblotting, increased approximately 1.9- and 1.3-fold, respectively, in mice fed a diet containing 0.7% (w/w) BHA for 7 days. The level of MSP23 mRNA in these tissues also increased more than 2-fold after mice were fed BHA, suggesting that the induction of MSP23 is controlled at the transcription level. Immunostaining of the small intestine shows that MSP23 is expressed mainly in the columnar epithelial cells. The induction of MSP23 may be important to protect the cells and tissues against toxic electrophiles and reactive oxygen species.

Animals↗
PubMed2000

Inhibitory effects of rosmarinic acid on the proliferation of cultured murine mesangial cells.

BACKGROUND: Rosmarinic acid is a phenolic compound widely distributed in Labiatae herbs such as rosemary, sweet basil, and perilla, which are frequently used with meat and fish dishes in Western and Asian countries. In the present study we investigated the effects of rosmarinic acid on cultured murine mesangial cell proliferation. METHODS: Cultured murine mesangial cells were stimulated by growth factors with or without rosmarinic acid, and [(3)H]thymidine incorporation was measured in regard both to signal transduction and to cell cycle dependency. In other experiments, mRNA extracted from the cells was analysed by Northern blotting. RESULTS: Rosmarinic acid inhibited the cell proliferation induced by platelet-derived growth factor (PDGF) (P<0.01; IC(50) values, 1.4 microg/ml) or tumour necrosis factor-alpha (P<0.01; IC(50) values, 3. 8 microg/ml), and these effects involved both the G(0)/G(1) and G(1)/S phases of the cell cycle. Rosmarinic acid also suppressed the mRNA expressions of PDGF and c-myc in PDGF-stimulated mesangial cells. CONCLUSIONS: Rosmarinic acid inhibits cytokine-induced mesangial cell proliferation and suppresses PDGF and c-myc mRNA expression in PDGF-stimulated mesangial cells. Rosmarinic acid in Labiatae herbs might be a promising agent to prevent mesangial cell proliferation.

Animals↗
PubMed2000

Interleukin 12 induces crescentic glomerular lesions in a high IgA strain of ddY mice, independently of changes in IgA deposition.

BACKGROUND: Our recently established high immunoglobulin (Ig)A inbred strain (HIGA) of ddY mice showed constantly high serum IgA levels, progressive mesangial sclerosis accompanied by IgA deposits, and elevated renal expression of transforming growth factor (TGF)-beta, mimicking IgA nephropathy. In the present study, we assessed the role of the immune system, especially of T cells, in this strain. METHODS: The in vitro production of interferon (IFN)-gamma, interleukin (IL)-4 and TGF-beta1 by splenic CD4+ T cells was assessed in HIGA mice at 14 and 28 weeks of age by comparison with age-matched C57BL/6 and BALB/c mice, T-helper (Th) 1, and Th2 prone controls respectively. Moreover, recombinant murine IL-12 was administered intraperitoneally to HIGA mice and serum IgA and renal lesions were analysed. RESULTS: The production of IFN-gamma by splenic CD4+ T cells was markedly upregulated in HIGA mice at both ages as compared with age-matched C57BL/6 and BALB/c mice. Although splenic CD4+ T cells from HIGA mice produced less IL-4 than those from BALB/c mice at both ages, the former produced significantly more IL-4 with age, which contrasted with the age-associated decrease in the latter. Moreover, TGF-beta1 production of these cells in HIGA mice was equal to or greater than that in the two groups of control mice at both ages. Daily intraperitoneal administration of IL-12 for 1 week significantly enhanced crescent formation with glomerular macrophage accumulation and interstitial cell infiltration, whereas it reduced the serum IgA level. CONCLUSIONS: In HIGA mice, Th1 is markedly upregulated from a young age and there is an age-associated Th2 increase with TGF-beta1 upregulation in helper T cells. The former may be related to the exacerbation of inflammatory renal lesions on IL-12 administration, while the latter may contribute to increased IgA production, leading to glomerular IgA deposition and progressive glomerulosclerosis in HIGA mice. The pathogenic role of T cell function and fluctuation of these subsets, especially the Th1/Th2 balance, is crucial to the immunopathological phenotype of the renal lesions in HIGA mice.

Animals↗
PubMed2000

Inhibition of the production of rat cytokine-induced neutrophil chemoattractant (CINC)-1, a member of the interleukin-8 family, by adenovirus-mediated overexpression of IkappaBalpha.

Cytokine-induced neutrophil chemoattractant (CINC)-1, a counterpart of the human growth-regulated gene product (GRO) of the interleukin-8 family, has been suggested to play critical roles as a mediator of inflammatory reactions with neutrophil infiltration in rats. NF-kappaB has been speculated to be involved in the production of CINC-1, since the NF-kappaB-binding domain is important for the CINC-1 promoter activity in several of our reporter assays. In the present study, we examined the effects of an overexpression of IkappaBalpha, a specific natural inhibitor of NF-kappaB, on CINC-1 production. For this purpose, we constructed two recombinant adenoviruses, AxCAIkappaBalpha and AxCAmutantIkappaBalpha, which express respectively wild IkBa and a mutated nondegradable IkappaBalpha in which serine residues 32 and 36 are replaced by alanine residues. Transfecting wild and mutant IkBa by these adenovirus vectors inhibited NF-kappaB activation and CINC-1 production, which were both caused by IL-1beta stimulation in the normal rat kidney epithelial cell line NRK-52E. We also showed that the nondegradable mutant IkappaBalpha was approximately 30 times more potent than the wild type in inhibiting CINC-1 production. These findings demonstrate that CINC-1 production with NF-kappaB activation is primarily regulated by non-phosphorylated IkBa in the cytoplasm.

Adenoviridae↗
PubMed2000

Intracellular degradation of histidine-rich glycoprotein mutants: tokushima-1 and 2 mutants are degraded by different proteolytic systems.

We reported the first case of a congenital histidine-rich glycoprotein deficiency (HRG Tokushima) in which substitution of Gly85 with Glu (G85E) in the first cystatin domain resulted in intracellular degradation and a low plasma level of HRG [Shigekiyo, T. et al. (1998) Blood 91, 128-133]. Recently, we identified the gene mutation of a second case of HRG deficiency as a Cys223 to Arg (C223R) mutation in the second cystatin domain. To investigate the molecular and cellular bases of these deficiencies, we expressed these HRG mutants in baby hamster kidney (BHK) cells. Pulse-chase experiments in the absence and presence of various proteinase inhibitors revealed that, while wild-type HRG was completely secreted during 4-h chase periods, both the G85E and C223R mutants were only partially secreted and primarily degraded within the cells. The intracellular degradation of the C223R mutant was almost completely inhibited in the presence of a proteasome inhibitor, lactacystin, carbobenzoxy-leucyl-leucyl-leucinal or N-acetyl-leucyl-leucyl-norleucinal, resulting in increased secretion of the C223R mutant, and thus implicating the proteasome system in this degradation process. In contrast, the sum of the amounts of the G85E mutant inside and outside the cells decreased during the chase periods even in the presence of the proteasome inhibitor, carbobenzoxy-leucyl-leucyl-leucinal or N-acetyl-leucyl-leucyl-norleucinal, although proteasome-specific inhibitor lactacystin and one of the cysteine protease inhibitors, E-64-d, prevented the intracellular degradation. These results suggested that intracellular degradation of G85E HRG occurred to some extent through a hitherto unknown mechanism. Similar studies involving recombinant mutants in which Gly85 or Cys223 was replaced with several other amino acids revealed that proteins with mutations leading to the destruction of the predicted b-sheet structure of the cystatin domains were eliminated by the intracellular quality control system.

Amino Acid Sequence↗
PubMed2000

NMR studies of model peptides of PHGGGWGQ repeats within the N-terminus of prion proteins: a loop conformation with histidine and tryptophan in close proximity.

The N-terminal region of the prion protein from human and mouse contains five tandem repeats with the consensus sequence of PHGGGWGQ. NMR studies were performed in water for two cyclic peptides, cyclo-[C(1)R(2)Q(3)P(4)H(5)G(6)G(7)S(8)W(9)G(10)Q(11)R(12)D(13)C(14)] (C1) and cyclo-[C(1)R(2)D(3)P(4)H(5)G(6)G(7)G(8)W(9)G(10)Q(11)P(12)H(13)G(14)G (15)G(16)W(17)G(18)Q(19)R(20)D(21)C(22)] (C2), which are cyclized by a disulfide bridge between the Cys residues at the N- and C-termini, and for their corresponding linear peptides (L1 and L2) which are formed by reduction. The patterns of the C(alpha)H chemical shift difference of these four peptide mimetics were very similar to those observed for the tandem repeats of human prion protein reported by other researchers. The medium-range NOE connectivities were found between the C(beta)H of the H5 and the proton of the W9 side chain for L1. The corresponding NOEs were also observed in H5-W9 and H13-W17 of L2 with ambiguity. These observations indicate that histidine (i) is in close proximity to tryptophan (i+4). d(alphaN) (i,i+2) NOE connectivities were observed between W9 and Q11 of L1 and L2, and d(NN) (i,i+1) NOE connectivities were also observed for G10-Q11 of L1 and L2 and for G18-Q19 of L2. Significantly lower temperature coefficients of amide proton chemical shifts were obtained for Q11 and Q19 of L2 and C2. Structure calculations for L1 showed that HGG(G/S)W and (G/S)WGQ adopt a loop conformation and a beta-turn, respectively. These results strongly suggest that the tandem repeats within prion protein adopt a non-random structure.

Amino Acid Sequence↗
PubMed2000

Osmium-metal coating device using hollow-cathode plasma CVD method.

A novel osmium-metal coating device for SEM observation has been developed to prevent negative charge build-up on specimens by applying the hollow-cathode low voltage discharge plasma chemical vapour deposition (CVD) method. The CVD method using the hollow-cathode offers the following advantages. (i) The method can deposit osmium-metal at very low discharge voltage that is as low as half of that of the planar parallel electrode method. Therefore, the method avoids damage due to ion bombardment during the coating process. (ii) The method can minimize the quantity of the OsO4 gas by introducing directly into the hollow-cathode. This feature is important to prevent the air pollution caused by the purged gas. (iii) A large coating area is guaranteed because the Os ion is filled in the hollow-cathode where the specimen is holed. (iv) The lower discharge voltage can be used by mixing Ar, N2 or air with the OsO4 gas as the environmental gas in the chamber. (v) The hybrid coating is also available by lining the appropriate metal material such as platinum (Pt) on the surface of the inside of the hollow-cathode. The method uses the plasma CVD of Os metal as well as the ion-sputter deposition of the lined metal.

Animals↗
PubMed2000

Construction of a diabody (small recombinant bispecific antibody) using a refolding system.

Diabodies are the recombinant bispecific antibodies (BsAbs), constructed from heterogeneous single-chain antibodies. Usually, diabodies have been prepared from bacterial periplasmic fraction using a co-expression vector (i.e. genes encoding two chains were tandemly located under the same promoter). Some diabodies, however, cannot be expressed as a soluble material owing to inclusion body formation, which limits the utilization of diabodies in various fields. Here we report an improved method for the construction of diabodies using a refolding system. As a model, a bispecific diabody binding to adenocarcinoma-associated antigen MUC1 and to CD3 on T cells was studied. One chain consisted of a VH specific for MUC1 linked to a VL specific for CD3 with a short polypeptide linker (GGGGS). The second was composed of a VL specific for MUC1 linked to a VH specific for CD3. The two hetero scFvs were independently obtained from intracellular insoluble fractions of Escherichia coli, purified, mixed stoichiometrically (at an equivalent molar ratio of 1:1) and refolded. The refolded two hetero scFv has a hetero-dimeric structure, with complete specificity for both target cells [i.e. MUC1 positive cells and CD3 positive lymphokine-activated killer cells with a T cell phenotype (T-LAK)]. Evaluation of the in vitro efficacy of T-LAK with the diabody by growth inhibition assay of cancer cells demonstrated maximum growth inhibition of cancer cells to reach approximately 98% at an effector:target ratio (E:T ratio) of 10, almost identical with that with anti-MUC1xanti-CD3 chemically synthesized BsAbs (c-BsAbs). This is the first report of the construction of a diabody using a refolding system.

Antibodies, Bispecific↗
PubMed2000

Smoking decreases alveolar macrophage function during anesthesia and surgery.

BACKGROUND: Smoking changes numerous alveolar macrophage functions and is one of the most important risk factors for postoperative pulmonary complications. The current study tested the hypothesis that smoking impairs antimicrobial and proinflammatory responses in alveolar macrophages during anesthesia and surgery. METHOD: The authors studied 30 smoking and 30 nonsmoking patients during propofol-fentanyl general anesthesia. Alveolar immune cells were harvested by bronchoalveolar lavage immediately and 2, 4, and 6 h after induction of anesthesia and at the end of surgery. The types of alveolar immune cell and macrophage aggregation were determined. The authors measured opsonized and unopsonized phagocytosis. Microbicidal activity was determined as the ability of the macrophages to kill Listeriamonocytogenes directly. Finally, RNA was extracted from harvested cells and cDNA was synthesized by reverse transcription. The expression of interleukin 1beta, 6, and 8, interferon gamma, and tumor necrosis factor alpha were measured by semiquantitative polymerase chain reaction using beta-actin as an internal standard. RESULTS: The fraction of aggregated macrophages increased significantly over time in both groups, whereas phagocytosis of opsonized and nonopsonized particles and microbicidal activity of alveolar macrophages decreased significantly. The changes, though, were nearly twice as great as in patients who smoked. Gene expression of all proinflammatory cytokines in alveolar immune cells except interleukin 6 increased 2- to 20-fold over time in both groups. The expression of interleukin 1beta, interferon gamma, and tumor necrosis factor alpha, however, increased only half as much in smokers as in nonsmokers. CONCLUSION: Smoking was associated with macrophage aggregation but markedly reduced phagocytic and microbicidal activity-possibly because expression of proinflammatory cytokines was reduced in these patients. Our data thus suggest that smokers may have a limited ability to mount an effective pulmonary immune defense after anesthesia and surgery.

Anesthesia, General↗
PubMed2000

Induction of cytokines and killer cell activities by cisplatin and 5-fluorouracil in head and neck cancer patients.

It has been suggested that certain antitumor agents stimulate antitumor immunity. In the present study, we examined whether cisplatin and 5-fluorouracil (5-FU) accelerate the antitumor host responses in head and neck cancer patients. Two groups of patients were studied, i.e. an untreated (UT) group and a treated, disease-free (TDF) group that received chemo-immunotherapy in combination with radiotherapy and operation. When peripheral blood mononuclear cells (PBMC) derived from head and neck cancer patients were treated with cisplatin or with 5-FU, interferon-gamma, tumor necrosis factor (TNF)-alpha, TNF-beta, interleukin (IL)-1beta, IL-6, IL-12 and IL-18 as well as killer cell activities were significantly induced in both groups. In this case, these activities induced by cisplatin in UT showed lower levels than those in TDF, whereas the activities induced by 5-FU in the UT group demonstrated almost similar levels to those in TDF. These activities were significantly inhibited by anti-asialo-GM1 antibody. Furthermore, cytokine levels in sera and killer activities of PBMC derived from the cancer patients were significantly increased after cisplatin administration. These findings suggest that cisplatin and 5-FU increase anticancer immunity mediated by induction of cytokines and killer cell activities in patients with head and neck cancer.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2000

Cytokine-inducing activity and antitumor effect of a liposome-incorporated interferon-gamma-inducing molecule derived from OK-432, a streptococcal preparation.

An interferon-gamma (IFN-gamma)-inducing molecule (OK-PSA) has been purified from OK-432 by an affinity chromatographic technique performed on cyanogen bromide-activated Sepharose 4B-bound TS-2 monoclonal antibody, which neutralizes IFN-gamma-inducing activity of OK-432. OK-PSA has striking anti-tumor activity in vivo and in vitro. In the current study, the liposomes were used to improve the delivery of the agent (OK-PSA) to effector cells and to increase the therapeutic effect. Significantly less OK-PSA encapsulated into liposomes (Lipo-OK-PSA) than OK-PSA alone (1/100 or less of OK-PSA alone) was required to induce IFN-gamma, tumor necrosis factor-alpha (TNF-alpha), TNF-beta, interleukin-1 beta (IL-1 beta), natural killer, and lymphokine-activated killer activities by human peripheral blood mononuclear cells and mouse spleen cells. Furthermore, higher levels of these activities were detected in peripheral blood mononuclear cells and mouse spleen cells treated with Lipo-OK-PSA than in those treated with OK-PSA. All of these activities induced by Lipo-OK-PSA were almost completely neutralized by anti-asialo-GM1 antibody and complement (p < 0.001). In in vivo experiments, Lipo-OK-PSA elicited striking anti-tumor activity on syngeneic Meth-A tumor-bearing and colon 26-bearing BALB/c mice and on salivary gland tumor-bearing nude mice far better than did OK-PSA. Furthermore, high levels of natural killer and lymphokine-activated killer activities and a significant increase in the number of cells positive for asialo-GM1, IFN-gamma, TNF-alpha, or IL-1 beta were detected in the spleen cells derived from the animals given Lipo-OK-PSA compared with those given saline. These findings clearly indicate that OK-PSA plays an important role in the anti-tumor efficiency of OK-432, and that, for the most part, liposome encapsulation of this molecule markedly accelerates its effect mediated by asialo-GM1-positive cells (mainly natural killer cells).

Adenocarcinoma↗
PubMed2000

The relationship between disc degeneration, facet joint osteoarthritis, and stability of the degenerative lumbar spine.

Degenerative processes in the disc and facet joints affect the stability of the motion segment. The exact relations among disc degeneration, facet joint osteoarthritis, and the kinematics of the motion segment are not well defined in the literature. Magnetic resonance imaging and functional radiography of the lumbar spine were analyzed to examine the relations among segmental instability, facet joint osteoarthritis, and disc degeneration in patients with degenerative disorders of the lumbar spine. Seventy consecutive patients (mean age, 46 years) had both magnetic resonance imaging and flexion and extension radiographs of the lumbar spine. The lumbar instability was classified into abnormal tilting on flexion, rotatory instability in the sagittal plane, and translatory instability. Translatory instability was subdivided into anterior, posterior, and anteroposterior translatory instability. Disc degeneration as seen on T2-weighted sagittal images was classified into five grades. Facet joint osteoarthritis as seen on axial T1-weighted images was divided into four grades. This study revealed that the kinematics of the lumbar motion segment are affected by disc degeneration and facet joint osteoarthritis. Abnormal tilting movement on flexion and anteroposterior translatory instability both had negative associations with facet joint osteoarthritis. However, anterior translatory instability was positively associated with disc degeneration and facet joint osteoarthritis. Rotatory instability in the sagittal plane and posterior translatory instability were not associated with disc degeneration and facet joint osteoarthritis.

Adolescent↗
PubMed2000

Anatomy of the iliolumbar ligament.

Information is lacking in the literature on the precise anatomy of the iliolumbar ligament and its individual differences. The morphologic pattern, length, and width of the iliolumbar ligament were determined in 56 embalmed lumbosacral spines from human cadavers. It was possible to classify the iliolumbar ligament into two groups: Type A (74 ligaments), in which anterior and posterior ligaments had separate courses; and Type B (32 ligaments), in which anterior and posterior ligaments moved together as one band. The angle of the posterior iliolumbar ligament in Type A was oriented significantly more posteriorly than that in Type B. The posterior iliolumbar ligament was significantly shorter and oriented more posteriorly in male anatomic specimens than in female ones.

Aged↗