Molecular genetic analysis of optineurin gene for primary open-angle and normal tension glaucoma in the Japanese population

PURPOSE: To determine whether mutations in the optineurin (OPTN) gene are associated with the incidence of primary open-angle glaucoma (POAG) and normal tension glaucoma (NTG) in the Japanese. METHODS: Eighty-nine unrelated Japanese patients with POAG and 65 unrelated patients with NTG were studied. Genomic DNA was extracted from leukocytes of the peripheral blood, and thirteen exons of the OPTN gene were amplified by polymerase chain reaction (PCR) and directly sequenced. RESULTS: Sequence alterations in exons 4 (His26Asp), 5 (Met98Lys), and 16 (Arg545Gln) were found. The His26Asp and Arg545Gln mutations were not detected in 100 ethnically matched controls. The frequency of the missense Met98Lys variant was higher in the POAG and NTG groups than in the control group (16.9% versus 5%, 15.4% versus 5%; P = 0.009 and P = 0.029, and odds ratio 3.85 and 3.45, respectively, for the dominant effect of the OPTN A allele). Polymorphisms in exons 4 and 12, and in introns 6 and 7 were also detected. CONCLUSIONS: The association of the allelic variation (Met98Lys) in the OPTN gene and the prevalence of POAG and NTG in unrelated Japanese patients suggest that they are involved in the pathogenesis of POAG and NTG.     

Differential expression of integrin subunits in DU-145/AR prostate cancer cells

We have established a clonal DU-145 prostate cancer cell line (DU-145/AR) stably transfected with androgen receptor cDNA. We investigated the expression of integrin subunits, adhesion to extracellular matrices, the invasion of DU-145/AR prostate cancer cells. The expression of various integrin subunits and adhesion to various extracellular matrices in DU-145, DU-145/Neo and DU-145/AR cells were examined. The haptoinvasion and the haptotactic migration of these cells were investigated using a Transwell cell culture chamber assay. DU-145/AR cells exhibited lower expression of alpha6 and beta4 integrin subunits and higher expression of alpha2 and alpha5 than DU-145 cells. DU-145/AR cells showed significantly lower adhesion to fibronectin, laminin-1 and laminin-5 than DU-145/ Neo cells, whereas DU-145/AR cells showed higher adhesion to type I and type IV collagen. Haptoinvasion of DU-145/AR cells into Matrigel/fibronectin-coated filter was significantly reduced as compared with DU-145/Neo or DU-145 cells, but there was no significant difference between DU-145/AR and control cells in the haptotactic migration to fibronectin. Dihydrotestosterone (DHT) inhibited the invasive ability of DU-145/AR cells. These results indicate that androgen receptor may play a role in the regulation of adhesion to the extracellular matrices and invasion of prostate cancer cells through influencing the expression of specific integrin subunits.     

Biomechanical considerations of fracture treatment and bone quality maintenance in elderly patients and patients with osteoporosis

Osteoporosis is a major public health problem that is characterized by low bone mass and structural deterioration of bone tissue, leading to bone fragility and an increased susceptibility to fractures of the hip, spine, and wrist. Poor bone quality in patients with osteoporosis presents the surgeon with difficult treatment decisions. Bone fracture repair has more pathways with combinations of bone formation mechanisms, which depend on the type of fracture fixation to be applied to achieve the desirable immobilization. There only may be one remodeling principle and in less than ideal conditions, mechanical and biophysical stimuli may provide effective augmentation of fracture healing in elderly patients. A different stimulus may limit its association to a specific healing mechanism. However, no matter which fixation method is used, an accurate reduction is a requisite for bone healing. Failure to realign the fracture site would result in delayed union, malunion, or nonunion. Therefore, a basic understanding of the biomechanics of osteoporotic bone and its treatment is necessary for clinicians to establish appropriate clinical treatment principles to minimize complications and enhance the patient's quality of life. We describe the biomechanical considerations of osteoporosis and fracture treatment from various aspects. First, bone structure and strength characterization are discussed using a hierarchical approach, followed by an innovative knowledge-based approach for fracture reduction planning and execution, which particularly is beneficial to osteoporotic fracture. Finally, a brief review of the results of several experimental animal models under different fracture types, gap morphologic features, rigidity of fixation devices, subsequent loading conditions, and biophysical stimulation is given to elucidate adverse mechanical conditions associated with different bone immobilization techniques that can compromise normal bone fracture healing significantly.     

Enucleation of renal cell carcinoma in von Hippel-Lindau disease using a microwave tissue coagulator

The patient was a 37-year-old man who had undergone left nephrectomy under the diagnosis of left renal cell carcinoma associated with von Hippel-Lindau (VHL) disease 4 years ago. Computed tomography (CT) revealed 3 individual tumors 20 mm, 13 mm and 9 mm in maximum diameter in the right kidney. All three renal tumors were enucleated with a microwave tissue coagulator (MTC) without renal pedicle clamping. There were no major complications related to nephron-sparing surgery such as postoperative bleeding, persistent urine leakage and deterioration of renal function. Our findings suggest that renal tumors with VHL disease can be enucleated using a MTC safely and successfully without damaging renal function.     

Effects of platelet release products on neutrophilic phagocytosis and complement receptors

INTRODUCTION: Platelets enhance leukocytic phagocytosis via the action of ATP and ADP in platelet release products (PRPr). The present study was designed to clarify the type of complements and complement receptors that are involved in phagocytosis activation by PRPr, ATP and ADP. MATERIALS AND METHODS: Human peripheral blood was used as the source of neutrophils and platelets. The supernatant of the platelet suspension after simulation was used as PRPr. The effects of PRPr, ATP, ADP, and other substances on neutrophilic phagocytosis, rosette formation and expression of several antigens were investigated. For the markers of neutrophilic phagocytosis and rosette formation, IgM-sensitized sheep red blood cells (SRBC) were treated with diluted human serum (EAC) or purified complements (C1, C4, C2 and C3) (EAC3b) followed by C3 inactivation (EAC3bi). The expressions of CD11b, CD11c, CD18, and CD35 were evaluated using a flow cytometer. RESULTS: Neutrophilic phagocytosis of EAC and EAC3bi was enhanced by PRPr, ATP, and ADP, whereas this phagocytosis activation was abolished by antibodies against CD11b and CD18. Neutrophil rosette formation with EAC3bi was increased by ATP and ADP. Flow cytometry revealed that the expressions of CD11b and CD35 on neutrophils were increased by PRPr, but not by ATP and ADP. The component in PRPr, responsible for the increase in expressions of these antigens, could not be identified. CONCLUSION: PRPr increases the neutrophilic phagocytosis of complement-coated particles through the action of ATP and ADP by increasing the binding avidity with iC3b, but not the number of Mac-1 (CD11b/CD18).     

Pulse dye densitometry using indigo carmine is useful for cardiac output measurement, but not for circulating blood volume measurement

BACKGROUND AND OBJECTIVE: We evaluated the validity of a newly developed pulse dye densitometer for indigo carmine for measuring cardiac output and circulating blood volume. METHODS: Measurements of cardiac output and circulating blood volume were performed with the indigo carmine densitometer during normovolaemia, hypovolaemia and hypervolaemia in nine mongrel dogs under general anaesthesia. The validity was evaluated by comparison of the values of cardiac output and circulating blood volume obtained by the thermodilution technique and the 51Cr-labelled red blood cell method, respectively. We also examined indigo carmine removal by continuous veno-venous haemofiltration after indigo carmine injection. RESULTS: There was good agreement between dye densitometer- and thermodilution-derived cardiac output (r = 0.885, P < 0.001). The bias and limits of agreement of these values were 0.09 and+/-1.07 L min(-1) (2 SD, n 22), respectively. The dye-densitometer-derived circulating blood volume was greater than that of the 51Cr-labelled red blood cell method, and both values showed weak agreement (r = 0.587, P < 0.027). The sieving coefficient of indigo carmine through continuous veno-venous haemofiltration was 0.34+/-0.06. CONCLUSIONS: These data indicate that indigo carmine densitometry is a reliable method for cardiac output determination, but it overestimates circulating blood volume, probably due to the transition of indigo carmine into the extravascular space in the systemic circulation.     

Th1 and Th2 CD4+ T cells and Tc1 and Tc2 CD8+ T cells of patients with Wegener's granulomatosis

A Th1/Th2 cytokine imbalance with a predominance of Th1 cytokines has been suggested to be of pathogenic importance in Wegener's granulomatosis. To evaluate the role of Th1/Th2 cytokines in Wegener's granulomatosis, the subsets of Th1, Th2, Tc1 and Tc2 cells from patients with active Wegener's granulomatosis were examined by intracellular cytokine flow cytometry. The population of Tc1 cells (72.0 +/- 14.4 per cent) in Wegener's granulomatosis was significantly increased compared with Tc1 cells (37.3 +/- 14.6 per cent) in control (p<0.05). Th1, Th2 and Tc2 cells in Wegener's granulomatosis were not significantly increased compared with the control cells. These results indicate that the predominance of Tc1 cells might contribute to the mechanism of the pathogenesis of Wegener's granulomatosis.     

Induction and characterization of cutaneous lymphocyte antigen on natural killer cells

Cutaneous lymphocyte antigen (CLA) has been reported to be expressed mainly by memory/effector T lymphocytes infiltrating inflammatory skin lesions and cutaneous T-cell lymphoma. It has been suggested that CLA is a specific homing receptor, facilitating the T-cell migration into skin lesions, and also an indicator of the skin-homing T-cell subset. In the present study, we investigated the expression of CLA in natural killer (NK) cells defined phenotypically as surface CD3- and CD56+ cells in peripheral blood. CLA was definitely expressed on CD3-CD56+ cells at a level comparable to CD3+ cells in peripheral blood of normal Japanese volunteers. After in vitro stimulation of peripheral blood mononuclear cells with interleukin 2 (IL-2) and IL-12, there was a significant increase in the number and percentage of CLA+ NK cells but not CLA+ T cells (P < 0.01). To analyse the characteristics of CLA expressed by NK cells, we investigated a CLA+ NK-leukaemia cell line, NK-YS, established from a patient with NK leukaemia/lymphoma with skin infiltration. In the in vitro study, the CLA-expressing NK-leukaemic cell line bound to E-selectin-transfected cells and was inhibited by HECA 452 antibody or neuraminidase treatment of leukaemic cells. These findings suggest that CLA expressed by NK cells is a homing receptor for the E-selectin molecule and may explain skin infiltration by NK cells and NK lymphoma cells analogous to T cells. An NK-cell subset expressing CLA must play an important role in host defence against microorganisms and neoplasms in skin lesions.