Nagoya Journal of Medical Science

To study the regulation of [Na+]i and [Ca2+]i during myocardial ischemia/reperfusion, [Na+]i and [Ca2+]i were measured simultaneously using guinea pig ventricular myocytes which were dual-loaded with SBFI/AM and fluo-3/AM. It was suggested that: (1) [Na+]i increased during metabolic inhibition (MI: 3.3 mM amytal and 5 μM CCCP) by both the activated Na+ influx via Na+/H+ exchange and the suppressed Na+ extrusion via the Na+/K+ pump; (2) Na+/Ca2+ exchange was inhibited during MI, causing the dissociation between [Na+]i and [Ca2+]I; (3) Na+/Ca2+ exchange could be reactivated by energy repletion, resulting in a significant increase in [Ca2+]i, Furthermore, a Ca2+ influx via the reverse-mode of Na+/Ca2+ exchange may play a key role in the mechanism of Ca2+ overload on reoxygenation; and (4) cell contracture during MI was related to rigor due to energy depletion, while cell contracture after energy repletion was likely to be related to Ca2+ overload.

The catalytic subunit of human DNA polymerase (pol) δ, p125, was expressed in recombinant baculovirus-infected insect cells, separated from a baculovirus-encoded DNA polymerase, and was purified to homogeneity by affinity trapping with a histidine-octapeptide at the C-terminus of p125 as the ligand. Purified p125 showed DNA polymerase activity resembling conventionally purified calf thymus pol δ. However, the two differed in four ways: 1) the specific activity of recombinant p125 was one quarter of the calf thymus pol δ; 2) the recombinant p125 was relatively resistant to aphidicolin; 3) the apparent Km for dTTP of the recombinant p125 was estimated at 33 µM, 15-fold the value for calf thymus pol δ; and 4) the recombinant p125 was not stimulated by recombinant PCNA, while activity of calf thymus pol δ increased 150-fold in response. Furthermore, PCNA did not stimulate either the p125 incubated with p50, a small subunit of pol δ, or co-expressed with p50 in insect cells. The full length recombinant p125 migrated slightly faster than pol δ from human cell lines, Jurkat or HeLa, upon SDS-polyacrylamide gel electrophoresis, suggesting a post-translational modification. The results indicate that in vivo assembly of the fully active complex of pol δ requires factors in addition to p125 and p50 subunits, and/or a post-translational modification of p125.

Objective Cold intolerance, cold induced peripheral vasospasm, Raynaud’s phenomenon, livedo reticularis and immunoglobulin deposition in the skin are often encountered clinical and laboratory findings in patients with primary fibromyalgia (FM). These findings are suggestive of vascular injury.
Methods
Eighty patients (4 male, 76 female) with fulfilling primary FM criteria (FM (+) patient group), 60 patients (3 male, 57 female) with chronic musculoskeletal complaints but without FM (FM (–) patient control group) and 40 healthy volunteers (1 male, 39 female) without musculoskeletal complaints (healthy control group) were enrolled in this cross-sectional study. The study was carried out in two steps. In the first step, the clinical findings, routine laboratory tests, autoantibodies and radiological findings were investigated. The second step were consisted of the laboratory investigations of thrombomodulin and fibronectin as the mediators indicating vascular injury and proinflammatory cytokines in FM patients with Raynaud’s phenomenon and/or livedo reticularis and in control groups.
Results
There were no differences between study and control groups with regard to laboratory, radiological and immunological (ANA, AntidsDNA, ENA, anticardiolipin IgG and IgM) results. No statistically significant differences were found in the levels of proinflammatory cytokines between FM (+) patient group and control groups (p > 0.05). Thrombomodulin was also shown statistically insignificant difference between FM (+) patient group and control groups (p > 0.05). However, fibronectin, another mediator of vascular injury, was higher in FM (+) patient group and the differences between FM (+) patients and each control groups were statistically significant (p < 0.0001).
Conclusion
Our results were suggestive of the presence of a non-immunological vascular injury in FM patients with Raynaud’s phenomenon and/or livedo reticularis

An 8-year-old girl died of sepsis due to staphylococcal infection one year and 8 months after Bacille Calmette-Guerin (BCG) revaccination. Two months after the vaccination in accordance with the school health program, she was hospitalized with a high fever, skin rash over the face and lower limbs, and leukopenia. Her clinical and laboratory pictures were not compatible with those of any established type of immunodeficiency. The polymerase chain reaction (PCR) test for M. tuberculosis complex was positive for bone marrow, pleural fluid, and peripheral blood. The strain recovered from a mycobacterial culture of the blood was identical to the BCG strains with which the patient was vaccinated, based on restriction fragment length polymorphism (RFLP) and a pulse-field gel electrophoresis (PFGE) analyses of DNA. She developed finally a lung abscess due to staphylococcal septicemia, which was the direct cause of her death.

Docetaxel (DOC) plus cisplatin (CDDP) is a novel combination chemotherapy for the treatment of advanced non-small-cell lung cancer (NSCLC). We investigated the combined effect of DOC with CDDP in sequence and the reverse schedule for NSCLC cell lines EBC-1 (squamous cell carcinoma) and RERF-LCMS (adenocarcinoma) using an MTT assay and an improved isobologram method. The results showed that the combination of DOC and CDDP in human lung cancer cell lines was antagonistic. To investigate the possible mechanism of the antagonistic effect, we focused on the cell cycle perturbation and the inhibition of apoptosis fractions induced by chemotherapeutic agents. Pretreatment of CDDP significantly blocked the following DOC-induced apoptosis fraction. Therefore we consider that the suppression of apoptosis could be one of the mechanisms for antagonistic effects of combination chemotherapy of DOC and CDDP.