Life-Long Wheel Running Attenuates Age-Related Fiber Loss in the Plantaris Muscle of Mice: a Pilot Study.

The purpose of this study was to investigate whether long-term wheel running would attenuate age-related loss of muscle fiber. Male ICR mice were divided into young (Y, n=12, aged 3 months), old-sedentary (OS, n=5, aged 24 months), and old-exercise (OE, n=6, aged 24 months) groups. The OE group started spontaneous wheel running at 3 months and continued until 24 months of age. Soleus and plantaris muscles were fixed in 4% paraformaldehyde buffer. The fixed muscle was digested in a 50% NaOH solution to isolate single fiber and then fiber number was quantified. The masses of the soleus and plantaris muscles were significantly lower at 24 months than at 3 months of age, and this age-related difference was attenuated by wheel running (P<0.05). Soleus muscle fiber number did not differ among the groups. In the plantaris muscle, the fiber number in the OS group (1 288±92 fibers) was significantly lower than in the Y group (1 874±93 fibers), and this decrease was attenuated in the OE group (1 591±80 fibers) (P<0.05). These results suggest that age-related fiber loss occurs only in the fast-twitch fiber-rich muscle of mice, and that life-long wheel running exercise can prevent this fiber loss.

Percutaneous shunt vessel embolisation with Amplatzer vascular plugs II and IV in the treatment of dogs with splenophrenic shunts: four cases (2019-2022).

OBJECTIVES: To describe the treatment of four dogs with splenophrenic shunts using percutaneous shunting vessel embolisation with Amplatzer vascular plugs II and IV and provide information on their clinical outcomes. MATERIALS AND METHODS: Dogs with splenophrenic shunts treated at a veterinary hospital from January 2019 to December 2022 were identified through a medical record search. RESULTS: Six dogs with splenophrenic shunts were identified. Two dogs were excluded because they were treated with laparoscopic surgery. Four underwent percutaneous shunting vessel embolization with Amplatzer vascular plugs and were included in the case series. A sheath was placed in the left external jugular vein and a balloon catheter was advanced to the shunting vessel under fluoroscopy. Portal vein pressure was confirmed to be within an acceptable range during temporary balloon occlusion. Based on preoperative CT angiography and intraoperative contrast examination, Amplatzer vascular plugs II were selected for two dogs and IV were selected for two dogs. Under fluoroscopy, the plug was deployed into the shunting vessel, and angiography confirmed occlusion. In all cases, the increase in portal pressure after temporary occlusion was within the acceptable range, and complete occlusion of blood flow was possible with a single plug. There were no major procedure-related complications. No dogs developed post-ligation seizures or signs of portal hypertension. In addition, improvements in ammonia values were observed in all cases. CLINICAL SIGNIFICANCE: Percutaneous splenophrenic shunt embolisation using Amplatzer vascular plugs II and IV is technically feasible in dogs, and assessed by intra-procedure angiography, a single plug completely obstructed blood flow in all dogs. Based on the literature search, this is the first report describing Amplatzer vascular plugs for the treatment of splenophrenic shunts.

Glycogen synthase kinase-3beta and p38 phosphorylate cyclin D2 on Thr280 to trigger its ubiquitin/proteasome-dependent degradation in hematopoietic cells.

Cyclin D2 plays an important role in regulation of hematopoietic cell proliferation by cytokines and is implicated in oncogenesis of various hematopoietic malignancies. However, mechanisms regulating cyclin D2 stability and its expression level have remained to be known. Here, we demonstrate that interleukin-3 signaling stabilizes cyclin D2 by inhibition of glycogen synthase kinase-3beta (GSK3beta) through Janus kinase2-dependent activation of phosphatidylinositol 3'-kinase (PI3K)/Akt signaling pathway in hematopoietic 32Dcl3 cells. On the other hand, osmotic stress was shown to induce a rapid proteasomal degradation of cyclin D2, which was mediated by activation of p38. GSK3beta and p38 was demonstrated to phosphorylate cyclin D2 on Thr280 in vitro, while a cyclin D2 mutant with this residue substituted with Ala was found to be resistant to ubiquitination and proteasome-dependent degradation in 32Dcl3 cells. Inhibition of the PI3K pathway or induction of osmotic stress also caused a rapid proteasomal degradation of cyclin D2 in primary leukemic or myeloma cells. These results indicate that cyclin D2 expression in normal and malignant hematopoietic cells is regulated by ubiquitin/proteasome-dependent degradation that is triggered by Thr280 phosphorylation by GSK3beta or p38, which is induced by inhibition of the PI3K pathway or by osmotic stress, respectively.

Rottlerin synergistically enhances imatinib-induced apoptosis of BCR/ABL-expressing cells through its mitochondrial uncoupling effect independent of protein kinase C-delta.

Although the BCR/ABL tyrosine kinase inhibitor imatinib is highly effective for treatment of chronic myeloid leukemia (CML) and Philadelphia-chromosome positive acute lymphoblastic leukemia (ALL), relapse with emerging imatinib-resistance mutations in the BCR/ABL kinase domain poses a significant problem. Here, we demonstrate that rottlerin, a putative protein kinase C-delta (PKCdelta)-specific inhibitor, acts synergistically with imatinib to induce apoptosis of BCR/ABL-expressing K562 and Ton.B210 cells. However, rottlerin inhibited neither PKCdelta nor BCR/ABL in these cells. On the other hand, rottlerin, previously characterized also as a mitochondrial uncoupler, transiently but significantly reduced mitochondrial membrane potential and gradually induced mitochondrial membrane permeabilization. Moreover, two other mitochondrial uncouplers, FCCP and DNP, very similarly induced apoptosis of BCR/ABL-expressing cells in a synergistic manner with imatinib. Imatinib synergistically enhanced mitochondrial membrane permeabilization induced by mitochondrial uncouplers, which led to release of cytochrome c into the cytoplasm and activation of caspases-3 and -9. Rottlerin also enhanced the cytotoxic effect of imatinib in leukemic cells from patients with CML blast crisis and Ph-positive ALL or a cell line expressing the imatinib-resistant E255K BCR/ABL mutant. The present study indicates that rottlerin synergistically enhances imatinib-induced apoptosis through its mitochondrial uncoupling effect independent of PKCdelta and may contribute to the development of new treatment strategy to overcome the imatinib resistance and to cure the BCR/ABL expressing leukemias.

CBP/catenin antagonist safely eliminates drug-resistant leukemia-initiating cells.

CREB-binding protein (CBP) and p300 are highly homologous transcriptional coactivators with unique, non-redundant roles that bind a wide array of proteins, including catenins-ß and γ. ICG-001 is a small-molecule inhibitor that specifically inhibits the CBP/catenin interaction. Importantly, ICG-001 does not inhibit the p300/catenin interaction. We demonstrate that specifically inhibiting the interaction between CBP and catenin with ICG-001 results in the differentiation of quiescent drug-resistant chronic myelogenous leukemia-initiating cells (CML LICs), thereby sensitizing them to BCR-ABL tyrosine kinase inhibitors, for example, Imatinib. Using ICG-001 in a NOD/SCID/IL2Rγ(-/-) mouse model of engrafted human chronic myelogenous leukemia, we now demonstrate the complete elimination of engrafted leukemia after only one course of combined chemotherapy. Combination-treated animals live as long as their non-engrafted littermates. Results from these studies demonstrate that specifically antagonizing the CBP/catenin interaction with ICG-001 can eliminate drug-resistant CML LICs without deleterious effects to the normal endogenous hematopoietic stem cell population.

Incidental Chylous Ascites at the Time of Cesarean Section.

Chylous ascites has multiple etiologies including malignancies, liver cirrhosis, intraperitoneal infections, and trauma. It is rarely reported in pregnancy. We report a case of chylous ascites noted at the time of cesarean section performed at 35 weeks of gestation on a patient with preeclampsia and suspected placental abruption. The diagnosis and treatment of chylous ascites as well as the pregnancy outcome are presented. A literature review of chylous ascites in pregnancy is discussed as well.

Blepharismins, produced by the protozoan, Blepharisma japonicum, form ion-permeable channels in planar lipid bilayer membranes.

Blepharismins are polycyclic quinones found in the pigment granules of the ciliated protozoan, Blepharisma. Exposure to purified blepharismins results in lethal damage to several other ciliates. We here report that, at cytotoxic concentrations, blepharismins formed cation-selective channels in planar phospholipid bilayer membranes. The channels formed in a diphytanoylphosphatidylcholine bilayer had a K(+)/Cl(-) permeability ratio of 6.6:1. Single channel recordings revealed the conductance to be quite heterogeneous, ranging from 0.2 to 2.8 nS in solutions containing 0.1 M KCl, possibly reflecting different states of aggregation of blepharismin. Our observations suggest that channel formation is a cytotoxic mechanism of blepharismin's action against predatory protozoa.

CD7+ near-tetraploid acute myeloblastic leukemia M2 with double t(8;21)(q22;q22) translocations and Aml1/ETO rearrangements detected by fluorescence in situ hybridization analysis.

Near-tetraploidy is a rare cytogenetic abnormality observed in acute myeloblastic leukemia (AML). It was recently suggested that near-tetraploid AML may be associated specifically with t(8;21)(q22;q22). We report here a new case of near-tetraploid AML with double t(8;21)(q22;q22) translocations. A 61-year-old woman was admitted to our hospital because of high fever and pancytopenia. Her bone marrow was markedly hypercellular, with 72% giant and bizarre myeloblasts. These myeloblasts were positive for CD7, CD13, CD19, CD33, CD34, and HLA-DR but negative for CD2 and CD56. The patient's disease was diagnosed as the M2 subtype of AML (by French-American-British classification). Chromosome analysis revealed a karyotype of 45,X,-X, t(8;21) (q22;q22)[1]/90,XXX,-X,t(8;21)(q22;q22)x2,-9[7]/46,XX[12]. Fluorescence in situ hybridization (FISH) analysis with an AML1/ ETO probe detected 4 fusion signals on 2 der(8)t(8;21) and 2 der(21)t(8;21) in near-tetraploid cells. Reverse transcription-polymerase chain reaction analysis revealed the presence of the AML1/ETO fusion transcript. These findings suggested that near-tetraploidy may be a secondary genetic change originating from a diploid clone with t(8;21) and that 2 AML1/ETO fusion genes are generated on the der(8)t(8;21) in near-tetraploid clones. Consideration of the other reported cases suggests that the immunophenotypes of near-tetraploid AML with double t(8;21) are heterogenous, but it is possible that t(8;21)-AML with expression of CD2 or CD7 may be associated with a secondary clonal evolution to near-tetraploidy.

Deletion of 16q11 is a recurrent cytogenetic aberration in acute myeloblastic leukemia during disease progression.

Abnormalities of chromosome 16 other than inv(16)(p13q22), t(16;16)(p13;q22), and del(16)(q22) have not been fully characterized in acute myeloblastic leukemia (AML) and myelodysplastic syndrome (MDS). We report here the first case of AML with del(16)(q11) as a sole abnormality. A 53-year-old woman was initially diagnosed as MDS, refractory anemia with excess of blasts in transformation with normal karyotype. After sixteen months, the disease progressed to overt AML-M1. Myeloblasts were positive for CD13, CD33, and CD34, but negative for HLA-DR. Chromosome analyses of the bone marrow cells showed 46,XX,del(16)(q11) in all metaphase spreads. Multicolor spectral karyotyping also confirmed that del(16)(q11) was not derived from a cryptic translocation, but a simple deletion. Our results, together with three previously reported cases, suggest that del(16)(q11) may be one of the recurrent aberrations in AML and that it could be associated with clonal evolution or disease progression.

A morphological study of age changes in adult human auricular cartilage with special emphasis on elastic fibers.

OBJECTIVE: It is well known that the size of the human auricle increases after it has finished development. The reason why the size of the human auricle continues to enlarge until advanced age after reaching adulthood was investigated by observation of the ultrastructure of elastic fibers in human auricular cartilage. METHODS: A total of 1958 subjects (966 males and 992 females) were classified into 18 age groups from 0 to 5 years up to 85 years and above by 5-year intervals. Ear length, ear width, and length of ear attachment were measured with calipers. Human auricular cartilage was obtained from 26 subjects (16 males and 10 females) aged 14 to 79 years, stained by orcein, and examined by light and electron microscopy. RESULTS: Each item of measurement of human auricular size increased significantly with age in both males and females. On morphological examination by light and electron microscopy after orcein staining, elastic fibers in the cartilage were almost homogeneous in diameter and surrounded the cartilage lacuna in bundle-like fashion in young persons, whereas those in elderly persons were heterogeneous in thickness and had many fragments surrounding the territorial matrix. In elderly persons, collagen-like fibers and small vesicles with heterogeneous electron density were frequently observed near elastic bundles around the territorial matrix. CONCLUSION: Structural changes of auricular cartilage associated with morphological age changes of elastic fibers may be one of the causes of expansion of the auricle after reaching adulthood.

Development of the human lateral vestibular nucleus: a morphometric evaluation.

The development of the human lateral vestibular nucleus was studied on serial sections of the brain of 8 fetuses and neonates at 12-40 weeks of gestation, an infant at 2 months of age and an adult of 63 years using a microscope with a drawing tube and an image-analysing computer system. A morphometric analysis revealed that the lateral vestibular nucleus, whose neurons were distinguished from glia after 16 weeks of gestation, divided cytoarchitectonically into the medial and the lateral subnuclei at 21 weeks of gestation onwards, and showed the moderate development in terms of the columnar length and volume, neuronal size and neuropil.

Immunochemical analysis of a photoreceptor protein using anti-IP3 receptor antibody in the unicellular organism, Blepharisma.

The blepharismin-200 kD protein complex of the ciliated protozoan Blepharisma is a novel type of photosensor responsible for the step-up photophobic response of the cell. In immunoblotting assays, the 200 kD protein is weakly cross-reacted with anti-inositol triphosphate receptor antibody (anti-IP3 R antibody). Indirect immunofluorescence assays show that the pigment granules in which the blepharismin-200 kD protein complex is localized are labelled by anti-IP3 R antibody. When the anti-IP3 R antibody or antisense oligonucleotide for IP3 receptor is introduced into the living cells of Blepharisma, both the photosensitivity of the cells and content of blepharismin-200 kD protein are reduced. The results suggest that the photoreceptor 200 kD protein is possibly an IP3 receptor-like protein.

Evaluation of expiratory effort on dysphonic patients on increasing vocal intensity.

It is conceivable that the subjects who have phonatory disorders, in comparison with normal individuals, exert a greater expiratory effort when phonating loudly. Furthermore, we presume that the extent and pattern of the changes in the expiratory effort for increasing vocal intensity may vary according to the types of laryngeal lesions. To prove these hypotheses, we investigated the changes in expiratory effort for increments of the vocal intensity by measuring the expiratory lung pressure. The subjects included 10 each of normal controls, patients with Reinke's edema, and those with recurrent laryngeal nerve paralysis. For the normal controls, the increase in vocal intensity was achieved by slightly increasing the expiratory lung pressure. The patients with Reinke's edema showed a greater increase in expiratory lung pressure, as compared with the normal group. The patients with recurrent laryngeal nerve paralysis exhibited greater expiratory effort with extreme increases in airflow than normal group for louder phonation. It was concluded that the subjects who have phonatory disorders, in comparison with normal individuals, require a greater expiratory effort. This phonatory function test with an increase in voice intensity made the aerodynamic pathologic condition clearer.

Demonstration of the nasal septal branches of the sphenopalatine artery by use of a new intravascular injection method.

We injected a new injection material into the external carotid artery using a new method, which led to the successful demonstration of the nasal septal branches of the sphenopalatine artery in human cadavers. The result shows the trunk of the artery divided into three main branches, the upper two of which run toward Little's area. We believe that the knowledge of septal branches, shown in a photograph, is very useful, not only for nasal treatment, but also for anatomic demonstration to students.

Use of digital subtraction fluoroscopy to diagnose radiolucent aspirated foreign bodies in infants and children.

OBJECTIVES: Most tracheobronchial foreign bodies in children are radiolucent, and accurate diagnosis of such foreign bodies is not always easy. This can result in delay of diagnosis or misdiagnosis of foreign body aspiration. We report the usefulness and pitfalls of use of digital subtraction fluoroscopy (DSF) to diagnose radiolucent aspirated foreign bodies in infants. METHODS: From 1991 through 1999, DSF was conducted for a total of 19 patients (ranged from 11 months to 4 years and 7 months in age (mean 1.8+/-0.9 years)) who were suspected to have radiolucent aspirated foreign bodies. Since DSF revealed abnormal findings in a trachea or main bronchus in 18 cases, inspection was performed for foreign body bronchofiberscopically. In the one remaining case, no abnormality was recognized on DSF, but since the symptoms at the time of onset strongly suggested aspirated foreign body, bronchofiberscopy was also performed. RESULTS: Foreign body was verified bronchoscopically in 13 of 19 cases, and all 13 (100%) had abnormal findings on DSF, including obstruction of the trachea in two, obstruction of the bronchial lumen in nine, and indistinct visualization of the bronchial lumen in two. Bronchial stenosis was verified bronchoscopically in five of the remaining six cases, including mucus plug in three, granuloma in one and mucosal edema in one case. All five patients (100%) had abnormal findings on DSF, including obstruction of the bronchial lumen in four and indistinct visualization of the bronchial lumen in one. In the one remaining patient with normal findings of DSF, no foreign body or pathological bronchial changes were noted. CONCLUSIONS: DSF was very sensitive in the diagnosis of foreign body aspiration and stenotic changes in the bronchial lumen. However, its diagnostic specificity for aspirated foreign body itself was not high (17%). Therefore, when abnormalities are found on DSF, we recommend to perform flexible bronchofiberscopy initially under general anesthesia via a tracheal tube. When a foreign body is verified, rigid ventilation bronchoscopy is successively performed to retrieve the foreign body.

Distribution of amyloid bodies in the aged human vestibulocochlear nerve.

We tried to elucidate the localization and distribution of amyloid bodies (Corpora amylacea) in the human vestibulocochlear nerve stained with luxol fast blue-periodic acid Schiff-hematoxylin using of a combination of an image analyzing computer system and a microscope fitted with a drawing tube. After having observed each section of the vestibulocochlear nerve from the brain stem to the fundus of the internal auditory meatus, we counted the numbers of amyloid bodies in three different parts for each of three corpses, and measured the areas. We found that amyloid bodies of the vestibulocochlear nerve are concentrated to the limiting glial portion of the nerve more than to the nerve parenchyma, and amyloid bodies are not seen in the vestibulocochlear nerve peripheral to the transitional zone. Our quantitative trial proved that the amyloid body was larger in the 8th decade than in the 6th or 7th decade of life.

Radiation therapy for metastatic spinal tumors.

PURPOSE: The results of radiation therapy for metastatic spinal tumors were evaluated in terms of pain relief, improvement of neurological impairment, and survival. MATERIALS AND METHODS: Between 1986 and 1995, 52 symptomatic patients with metastatic spinal tumors treated with radiation therapy were evaluated. The patients all received irradiation of megavoltage energy. Therapeutic efficacy was evaluated in terms of pain relief and improvement of neurological impairment. RESULTS: Pain relief was observed in 29 (61.7%) of 47 patients with pain. Therapy was effective for 17 (70.8%) of 24 patients without neurological impairment, and efficacy was detected in 12 (52.2%) of 23 patients with neurological impairment. Improvement of neurological symptoms was obtained in seven (25.0%) of 28 patients with neurological impairment. CONCLUSION: Radiation therapy was effective for pain relief in patients with metastatic spinal tumors. In patients with neurological impairment, less pain relief was observed than in those without impairment. Improvement of neurological impairment was restricted, but radiation therapy was thought to be effective in some cases in the early stage of neurological deterioration. Radiation therapy for metastatic spinal tumors contraindicated for surgery was considered effective for improvement of patients' activities of daily living.