Transformed phenotypes in long-term cultures persistently infected with bovine leukemia virus

Two cell lines, 3178 and FLK, were established respectively from calf form bovine lymphosarcoma and from fetal lamb kidney cells inoculated in vitro with bovine leukemia virus. These two cell lines persistently infected with bovine leukemia virus were maintained for more than 150 passages over three years. They exhibited the characteristics of transformed cell lines in vitro: 1) anchorage independence, 2) increased saturation density and decreased population doubling time, 3) increased uptake of 2-deoxy-D-glucose and 4) tumorigenicity in athymic nude mice.     

Inhibitory effects of macrophages against Marek's disease virus plaque formation in chicken kidney cell cultures.

Inhibition of plaque formation by Marek's disease virus (MDV) in chicken kidney cell cultures was investigated with the use of peritoneal exudate cells (PEC) from chickens. PEC from MDV-infected White Leghorn chickens inhibited the formation of MDV plaques, whereas the inhibitory effect of PEC from chickens vaccinated with herpesvirus of turkey (HVT) or PEC from normal chickens was very weak. However, PEC from either normal chickens or HVT-vaccinated chickens inhibited the MDV plaque formation in the presence of serum from MDV-infected chickens but not from normal or HVT-vaccinated chickens. The capacity of PEC to inhibit plaque formation was significantly reduced when PEC was treated with carrageenan but not with antithymus or antibursa cell serum. These results indicate that macrophages may have a role in protection against Marek's disease by reducing the number of MDV-infected cells and thereby decreasing the spread of the virus in vivo.     

Granulomatous hypophysistis associated with rathke's cleft cyst: a case report.

OBJECTIVE: The value of surgical intervention in the management of hypophysitis remains controversial. PATENT: We describe a 57-year-old man presenting with general fatigue persisting for three months. Endocrine examination revealed hypopituitarism and diabetes insipidus. Magnetic resonance (MR) imaging showed a dumbbell-shaped pituitary mass lesion without a cystic component. We partially removed the lesion via a transsphenoidal approach. Histological examination yielded the diagnosis of granulomatous hypophysitis associated with Rathke's cleft cyst. No deterioration of pituitary function was observed postoperatively. Twenty months after the surgery, the lesion has spontaneously regressed on MR imaging, and he is doing well with continuing replacement therapy of hydrocortisone, levothyroxine and desmopressin acetate. CONCLUSION: The diagnosis of hypophysitis, apart from typical lymphocytic hypophysitis, is difficult even with a surgical biopsy. Because a small specimen may lead to a diagnosis of non-specific hypophysitis, partial removal of the lesion is less invasive and recommended for preservation of the pituitary function.     

β3 Integrins Regulate Lymphocyte Migration and Cytokine Responses in Heart Transplant Rejection

Integrin alpha v beta 3 is important for cell survival, signaling and migration, particularly during angiogenesis and tumorigenesis, where it has been proposed as a therapeutic target. alpha v beta 3 is up-regulated following transplantation and beta 3 polymorphisms are associated with increased acute kidney rejection, suggesting that alpha v beta 3 may also play a role in transplant rejection. Here, using a model of allogeneic heart transplantation, we show that allograft survival is prolonged in beta 3 integrin-deficient (beta 3(-/-)) mice. This is associated with Th2-type immune responses and reduced T-cell infiltration into grafts and T cells from beta 3(-/-) mice show impaired adhesion and migration, consistent with a role for alpha v beta 3 in transmigration. These studies provide evidence that targeting beta 3 integrins impairs recruitment of effector cells and alters cytokine production, so prolonging graft survival. We also show that low doses of blocking antibodies against leukocyte function associated antigen-1 (LFA-1)/alpha L beta 2 and very late antigen-4 (VLA-4)/alpha 4 beta 1, when combined with deletion of beta 3, lead to long-term survival of allografts with no evidence of chronic rejection. Hence we provide strong mechanistic evidence supporting previous genetic studies, demonstrate the involvement of beta 3 integrins in both acute and chronic rejection and identify beta 3 as a new target for immunosuppressive therapy.     

Pregnanetriol in the Range of 1.2–2.1 mg/m2/day as an Index of Optimal Control in CYP21A2 Deficiency

Auxological data is the gold standard index of the therapeutic condition in CYP21A2 deficiency over a long-range period, whereas urinary pregnanetriol for 24 h (PT) is variable for a shorter-range period. Ideal PT levels in comparison with auxological data have not been reported. The main purpose of this study was to analyze ideal PT values as an index of optimal control for CYP21A2 deficiency. First, inter-daily fluctuation of PT was analyzed in one participant. PT levels were distributed over a wide range of 0.44–14.7 mg/day (n=42) in this participant, suggesting that the therapeutic condition should be judged by multiple PT samples. Second, the therapeutic periods of 15 participants with CYP21A2 deficiency were classified using auxological data and Cushing-like symptoms, and the PT levels were analyzed in each period retrospectively. The 95% confidence intervals for the means of the PT levels in the excessive, good and poor control periods were 0.03–1.25 (n=26), 1.23–2.09 (n=116), and 5.35–8.37 (n=72) mg/m²/day, respectively. In conclusion, 1.2–2.1 mg/m²/day of PT values can be used as an index of optimal control in CYP21A2 deficiency.     

The Range of 2.2–3.3 mg/gCr of Pregnanetriol in the First Morning Urine Sample as an Index of Optimal Control in CYP21 Deficiency

Auxological data are the gold standard indexes of the therapeutic conditions in patients with CYP21 deficiency over long-term periods, whereas urinary pregnanetriol (PT) for 24 h has been used as an index for short-term periods. We previously reported that the range of 1.2–2.1 mg/m²/day of PT for 24 h (24-h PT) could be used as an index of optimal control in patients with CYP21 deficiency. The purpose of this study was to analyze the range of PT in the first morning urine samples (morning PT) as an index of optimal control in patients with CYP21 deficiency. First, the therapeutic periods of 15 participants (aged 2 yr and 5 mo to 17 yr and 4 mo) were classified into excessive, good or poor control periods using auxological data and Cushing-like symptoms, and 24-h PT levels were analyzed in each period, retrospectively. The 95% confidence intervals for the means of 24-h PT levels in the excessive, good and poor control periods were 0.24–2.24 (n=25), 2.88–4.92 (n=114) and 13.26–21.28 (n=72) mg/gCr, respectively. Subsequently, 24-h PT and morning PT levels collected on the same day were analyzed for 14 participants (aged 9 mo to 29 yr and 8 mo). There was a significant correlation between the above two PT levels (n=25, p<0.0001). When the 24-h PT range of the good control period, 2.88–4.92 mg/gCr, was adjusted by the correlation, the ideal morning PT range became 2.15–3.34 mg/gCr. In conclusion, a morning PT in the range of 2.2–3.3 mg/gCr can be used as an index of optimal control in patients with CYP21 deficiency.     

Management and prognosis of cysts developed on long-term follow-up after Gamma Knife radiosurgery for intracranial arteriovenous malformations

BACKGROUND: Delayed cyst formation is a well-recognized complication after radiosurgery for intracranial AVM. The objective of the present study was the evaluation of the different management options for these lesions and the corresponding prognosis of patients. METHODS: Between 2000 and 2005, 12 patients with intracranial AVM initially treated by GKR were reevaluated at Tokyo Women's Medical University because of delayed cyst formation in the vicinity of the target area. There were 7 men and 5 women. The mean age of the patients was 31.8 years at the time of GKR and 41.1 years at the time of complication. The average period between treatment and diagnosis of the complication constituted 6.7 years. All AVMs had lobar location and showed complete angiographic obliteration after GKR. RESULTS: The most common neurological signs and symptoms at the time of cyst presentation were headache (10 cases) and seizures (4 cases). Two patients were asymptomatic. Three patients underwent surgery soon after the diagnosis of the cyst, whereas initial observation was done in another 9. Among the latter, 5 patients had to be treated surgically thereafter because of persistent or aggravated neurological symptoms associated with radiological cyst expansion. Four other patients, including both asymptomatic ones, are in stable condition without surgery. Follow-up after treatment of the cyst varied from 7 to 60 months (average, 34.3 months). All patients are in good condition. CONCLUSIONS: Although delayed formation of cysts after GKR for intracranial AVM should be considered as a complication of the radiosurgical treatment, it has a relatively good prognosis. Observation can be recommended as initial option for compensated and asymptomatic patients.     

Vimentin-Ser82 as a memory phosphorylation site in astrocytes

In astrocytes, the PGF(2alpha) or ionomycin treatment induces the phosphorylation at Ser38 and Ser82 of vimentin, a type III intermediate filament, by Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). We found here that vimentin phospho-Ser82 was dephosphorylated much slower than phospho-Ser38. Vimentin phospho-Ser38 was dephosphorylated quickly by purified PP1 catalytic subunit (PP1c) in vitro, whereas phospho-Ser82 was insensitive to PP1c. Because PP1c directly bound to vimentin through a VxF motif (Val83-Asp84-Phe85), the PP1c active site appeared to be unable to approach phospho-Ser82, leading to the prolongation of the phosphorylation at Ser-82. In astrocytes, PP1calpha was in vivo associated with vimentin filaments. The repetitive treatment by ionomycin at a short interval resulted in the sustained elevation of Ser82 phosphorylation, leading to the marked disassembly of vimentin filaments. Taken together, these results suggest that vimentin is a novel member of binding partner of PP1c in astrocytes, and vimentin-Ser82 may act as a memory phosphorylation site.