A perforin/granzyme-positive MDS-derived T cell line, K2-MDS, induces apoptosis in CD34+ cells through the fractalkine-CX3CR1 system

Fractalkine (CX3CL1) and its receptor CX3CR1 play an important role in natural killer (NK) cell- and cytotoxic T cell-mediated endothelium damage. Here we describe the cytotoxicity of myelodysplastic syndrome (MDS)-derived T cell line, K2-MDS, through the fractalkine-CX3CR1 system. K2-MDS cells induced apoptosis against CD34(+) cells from normal bone marrow (BM) in a direct cell contact manner. K2-MDS cells expressed perforin and granzyme B, but they lacked Fas ligand expression. A specific inhibitor for perforin, concanamycin A, blocked K2-MDS-dependent cytotoxicity. Furthermore, a CX3C-chemokine, fractalkine, was expressed in CD34(+) cells, and its receptor, CX3CR1, was expressed on K2-MDS cells. The neutralizing monoclonal antibody (MoAb) for fractalkine and soluble fractalkine significantly inhibited K2-MDS-dependent cytotoxicity. K2-MDS cells also induced the cytotoxicity against human umbilical cord endothelial cells (HUVECs) expressing fractalkine. These data indicate that K2-MDS may be a perforin-granzyme-positive T cell line that exerts a cytotoxic effect on CD34(+) cells mediated through the fractalkine-CX3CR1 system.     

Cognitive Frailty as a Risk Factor for Incident Disability During Late Life: A 24-Month Follow-Up Longitudinal Study

The journal of nutrition, health & aging - Association between cognitive frailty as identified by a new operational definition and incident disability in the community setting remains unclear....     

Modulation of M2-type pyruvate kinase activity by the cytoplasmic PML tumor suppressor protein

The promyelocytic leukemia (PML) tumor suppressor protein accumulates in PML nuclear bodies (PML-NBs), and can induce growth arrest, cellular senescence and apoptosis. PML has also been localized in the cytoplasm, although its function in this localization remains elusive. A general property of primary cancers is their high glycolytic rate which results from increased glucose consumption. However, the mechanism by which cancer cells up-regulate glycolysis is not well understood. Here, we have shown that cytoplasmic PML (cPML) directly interacts with M2-type pyruvate kinase (PKM2), a key regulator of carbon fate. PKM2 determines the proportion of carbons derived from glucose that are used for glycolytic energy production. Over-expression of PML-2KA mutant in the cytoplasm, which was generated by mutagenesis of the nuclear localization signals of PML, in MCF-7 breast cancer cells suppressed PKM2 activity and the accumulation of lactate. PKM2 exists in either an active tetrameric form which has high affinity for its substrate phosphoenolpyruvate (PEP) or a less active dimeric form which has low affinity for its substrate. Over-expression of PML-2KA suppressed the activity of the tetrameric form of PKM2, but not the dimeric form. Our findings suggest that cPML plays a role in tumor metabolism through its interaction with PKM2.     

Osteosarcoma occurring in osteogenesis imperfecta

We describe a case history of a 24-year-old male with osteogenesis imperfecta (OI) who developed osteosarcoma of the left thigh. High-dose ifosfamide therapy caused marked tumor regression of multiple lung metastases. Immunohistochemically, the tumor cells were diffusely positive for the p53 protein. Mutation of the p53 gene was not detected by direct genomic sequencing of exons 4–8. The radiographic characteristics, including irregularly distributed osteolytic lesions and cortical discontinuity, should not be confused with hyperplastic callus formation, a benign process. A biopsy is critical to establish the differential diagnosis between osteosarcoma and common hyperplastic callus formation in OI; however, it must be applied with great care.     

Genital herpes due to acyclovir-sensitive herpes simplex virus caused secondary and recurrent herpetic whitlows due to thymidine kinase-deficient/temperature-sensitive virus

Herpes simplex virus (HSV)-2 caused a genital ulcer in a 40-year-old allogenic stem cell recipient, and a secondary herpetic whitlow appeared during 2 months of acyclovir (ACV) therapy. Both genital ulcer, and whitlow were cured 3 months later, but 6 months after recovery the whitlow alone recurred. DNA of the genital, first, and recurrent whitlow isolates showed similar endonuclease digestion fragment profiles. The genital virus was ACV-sensitive, and the two whitlow isolates were ACV-resistant/thymidine kinase (TK)-deficient. The TK gene of the whitlow isolates had the same frame shift from the 274th amino acid and termination at the 347th amino acid due to the deletion of a cytosine at the 819th nucleotide. Because the temperature of the thumb is 33/34 degrees C or lower, the temperature sensitivity of the isolates were compared, and both whitlow isolates were significantly more temperature-sensitive (ts) at 39 degrees C than the genital isolate. The two whitlow isolates showed cutaneous pathogenicity in mouse ear pinna but not midflank, while the genital isolate was pathogenic at both sites, suggesting that temperature adaptation was an important element of pathogenicity in the whitlow. The virus populations of isolates of the genital, and first whitlow were examined by 31, and 82 clones, respectively, and the clones from genital, and whitlow isolates were ACV-sensitive, and -resistant, respectively, showing their homogeneity. The acyclovir-sensitive genital lesion had spread as a TK-deficient/ts herpetic whitlow during ACV treatment, and an apparently TK-deficient virus adapted to the local temperature might have caused the whitlow recurrence.     

Lack of mutagenicity of levofloxacin in lacZ transgenic mice

The mutagenic effects of levofloxacin in lacZ transgenicmice(Muta^TMMouse) have been investigated. Male Muta^TMMouse micewere administered levofloxacin i.p. at a dose of 300 or 600mg/kg. The higher dose corresponded to half the LD50 of thecompound in ddY strain mice. The mutant frequencies in the bonemarrow, liver (day 10 only), testis and sperm were examinedby the positive selection method for lacZ mutations on days1 and 10 after treatment Levofloxacin did not induce any statisticallysignificant increase in mutant frequency in any of the examinedtissues at either dose level or at either sampling time. Themutant frequency increases over the spontaneous values in thebone marrow, liver, testis and sperm were 1.0- to 1.2-fold,0.9-fold, 0.5- to 1.0-fold and 0.9- to 13-fold respectively.N-Ethyl-N-nitrosourea (100 mg/kg as a positive control), onthe other hand, induced significant increases in mutant frequenciesin both somatic cells (bone marrow and liver) and germ cells(testis and sperm) on day 10 after treatment. The mutagenicpotency for ENU was bone marrow >> liver testis sperm(50.1-fold, 3.4-fold, 2.9-fold and 23-fold respectively overthe spontaneous values). Levofloxacin was not mutagenic in thelacZ transgenic mice under the present experimental conditions. ¹To whom correspondence should be addressed. Tel: +81 3 5696 8294; Fax: +81 3 5696 8335; Email: itohsudk{at}daiichipharm.co.jp     

Developmental morphology of vascular and lymphatic capillaries in the working myocardium and Purkinje bundle of the sheep septomarginal band

The normal development of vascular and lymphatic capillaries in the right ventricular septomarginal band of the sheep heart was studied in 9 fetuses aged 60-143 days (term = 147 days), 14 lambs aged 1 day to 16 weeks, and 3 adults. Tissue was fixed by perfusion and examined with light and transmission electron microscopy. The septomarginal band is composed of working myocardium and a well-defined peripheral bundle of Purkinje cells. Vascular capillaries of the working myocardium were closely apposed to myocardial cells. By contrast, vascular capillaries of the Purkinje bundle were situated within the connective tissue sheath and septa, at variable distances from the Purkinje cells. After birth, the capillaries of the Purkinje bundle were also found in grooves and tunnels within the Purkinje strands. The ultrastructure of fetal vascular capillaries associated with myocardial and Purkinje cells was initially similar, and characterized by an abundance of synthetic organelles in endothelial cells and pericytes. However, after 115 days in utero, capillary endothelium with diaphragmed fenestrae, 40-60 nm in width, were observed within the Purkinje bundle. The fenestrae attained an average frequency of 1 per 11 capillary cross sections just before term, and this was maintained in lambs and adults. The ultrastructure of lymphatic capillaries, which were not observed in the septomarginal band until just before term, changed little during development.     

Right vertebral artery as the fourth branch of the aortic arch

The present report describes an anomalous case of the right vertebral artery arising as the last branch of the aortic arch identified in a 76-year-old Japanese male cadaver during dissection in the anatomical laboratory of Kanazawa Medical University. The aortic arch itself coursed normally but the right vertebral artery was uniquely situated at the fourth branch next to the brachiocephalic artery, the left common carotid artery, and the left subclavian artery. The anomalous right vertebral artery branched into the esophageal branch, the prevertebral branch, and the second right posterior intercostal artery, and finally entered the first costotransverse foramen at the thoracic region as it passed upward through the first to the seventh transverse foramina of the cervical vertebra. The left vertebral artery was normal. The development of the right vertebral artery may be described as follows: (i) the distal portion of the right dorsal aorta, which usually disappears, persisted and became united, via post-costal longitudinal anastomosis; (ii) the right dorsal aorta between the seventh and eighth intersegmental arteries lost its connection to the main structure; and (iii) the fusion of the originally paired dorsal aorta extended around the 11th segment, which was two segments away from the normal portion of the structure.