Prognostic factors associated with mortality before and during anti-tuberculosis treatment.

OBJECTIVE: To identify factors associated with death before the start of anti-tuberculosis treatment, and early and late during treatment, among adult Taiwanese with culture-positive pulmonary tuberculosis (PTB). METHOD: All adult culture-positive PTB patients in Taipei, Taiwan, were included in a retrospective cohort study in 2005-2010. RESULTS: Of 4438 patients (mean age 64.6 years, 70.6% male), 76.8% were successfully treated, 5.4% died before start of treatment, 9.0% died within 8 weeks of treatment initiation and 8.8% died >8 weeks after treatment initiation. After controlling for potential confounders, age ≥ 65 years and male sex were associated with higher risks of death at all time periods investigated. High school education or higher reduced the risk of death before the start of and during treatment, while unemployment increased the risk of mortality during treatment. Cavity on chest X-ray and positivity for acid-fast bacilli were associated with lower risk of mortality before the start of treatment. CONCLUSION: To lower mortality among adult culture-positive PTB patients, it is imperative for clinicians to maintain high awareness of TB and provide more intensive care early, especially for men, the elderly and people with lower socio-economic status (e.g., the unemployed and less educated).

Vaccinia virus envelope D8L protein binds to cell surface chondroitin sulfate and mediates the adsorption of intracellular mature virions to cells.

We previously showed that an envelope A27L protein of intracellular mature virions (IMV) of vaccinia virus binds to cell surface heparan sulfate during virus infection. In the present study we identified another viral envelope protein, D8L, that binds to chondroitin sulfate on cells. Soluble D8L protein interferes with the adsorption of wild-type vaccinia virions to cells, indicating a role in virus entry. To explore the interaction of cell surface glycosaminoglycans and vaccinia virus, we generated mutant viruses from a control virus, WR32-7/Ind14K (A27L(+) D8L(+)) to be defective in expression of either the A27L or the D8L gene (A27L(+) D8L(-) or A27L(-) D8L(+)) or both (A27L(-) D8L(-)). The A27L(+) D8L(+) and A27L(-) D8L(+) mutants grew well in BSC40 cells, consistent with previous observations. However, the IMV titers of A27L(+) D8L(-) and A27L(-) D8L(-) viruses in BSC40 cells were reduced, reaching only 10% of the level for the control virus. The data suggested an important role for D8L protein in WR32-7/Ind14K virus growth in cell cultures. A27L protein, on the other hand, could not complement the functions of D8L protein. The low titers of the A27L(+) D8L(-) and A27L(-) D8L(-) mutant viruses were not due to defects in the morphogenesis of IMV, and the mutant virions demonstrated a brick shape similar to that of the control virions. Furthermore, the infectivities of the A27L(+) D8L(-) and A27L(-) D8L(-) mutant virions were 6 to 10% of that of the A27L(+) D8L(+) control virus. Virion binding assays revealed that A27L(+) D8L(-) and A27L(-) D8L(-) mutant virions bound less well to BSC40 cells, indicating that binding of viral D8L protein to cell surface chondroitin sulfate could be important for vaccinia virus entry.

Cell surface proteoglycans are necessary for A27L protein-mediated cell fusion: identification of the N-terminal region of A27L protein as the glycosaminoglycan-binding domain.

We previously showed that vaccinia virus infection of BSC40 cells was blocked by soluble heparin, suggesting that cell surface heparan sulfate mediates vaccinia virus binding (C.-S. Chung, J.-C. Hsiao, Y. -S. Chang, and W. Chang, J. Virol. 72:1577-1585, 1998). In this study, we extended our previous work and demonstrated that soluble A27L protein bound to heparan sulfate on cells and interfered with vaccinia virus infection at a postbinding step. In addition, we investigated the structure of A27L protein that provides for its binding to heparan sulfate on cells. A mutant of A27L protein, named D-A27L, devoid of a cluster of 12 amino acids rich in basic residues, was constructed. In contrast to the soluble A27L protein, purified D-A27L protein was inactive in all of our assays, including binding to heparin in vitro, binding to heparan sulfate on cells, and the ability to block virus infection. These data demonstrated that the N-terminal region acts as a glycosaminoglycan (GAG)-binding domain critical for A27L protein binding to cells. Previously A27L protein was thought to be involved in fusion of virus-infected cells induced by acid treatment. When we investigated whether cell surface GAGs also participate in A27L-dependent fusion, our results indicated that soluble A27L protein blocked cell fusion, whereas D-A27L protein did not. Taken together, the results therefore demonstrated that A27L-mediated cell fusion is triggered by its interaction with cell surface GAGs through the N-terminal domain.

Primary retroperitoneal liposarcoma mimicking ovarian cancer: a case report.

Primary retroperitoneal liposarcoma is a rare malignancy comprising about only 0.1% of all cancers. It produces nonspecific symptoms and is often extensive when diagnosed. In this report, we present a case of a 68-year-old female patient who had a 29-kg retroperitoneal liposarcoma. Her early symptoms--including vague digestive disturbances, increasing abdominal girth and an abdominal mass, and clinical examinations such as sonography and computed tomography scan led to a preoperative diagnosis of ovarian cancer, until surgical and pathologic confirmation. Gross, radical resection of the tumor was successfully performed, and provided the most effective primary therapeutic approach. Histopathology revealed a mixed-type liposarcoma, with metastasis to the appendix. A poor prognosis was expected. Postoperative periodic follow-up was started to monitor for early detection of recurrence.

A27L protein mediates vaccinia virus interaction with cell surface heparan sulfate.

Vaccinia virus has a wide host range and infects mammalian cells of many different species. This suggests that the cell surface receptors for vaccinia virus are ubiquitously expressed and highly conserved. Alternatively, different receptors are used for vaccinia virus infection of different cell types. Here we report that vaccinia virus binds to heparan sulfate, a glycosaminoglycan (GAG) side chain of cell surface proteoglycans, during virus infection. Soluble heparin specifically inhibits vaccinia virus binding to cells, whereas other GAGs such as condroitin sulfate or dermantan sulfate have no effect. Heparin also blocks infections by cowpox virus, rabbitpox virus, myxoma virus, and Shope fibroma virus, suggesting that cell surface heparan sulfate could be a general mediator of the entry of poxviruses. The biochemical nature of the heparin-blocking effect was investigated. Heparin analogs that have acetyl groups instead of sulfate groups also abolish the inhibitory effect, suggesting that the negative charges on GAGs are important for virus infection. Furthermore, BSC40 cells treated with sodium chlorate to produce undersulfated GAGs are more refractory to vaccinia virus infection. Taken together, the data support the notion that cell surface heparan sulfate is important for vaccinia virus infection. Using heparin-Sepharose beads, we showed that vaccinia virus virions bind to heparin in vitro. In addition, we demonstrated that the recombinant A27L gene product binds to the heparin beads in vitro. This recombinant protein was further shown to bind to cells, and such interaction could be specifically inhibited by soluble heparin. All the data together indicated that A27L protein could be an attachment protein that mediates vaccinia virus binding to cell surface heparan sulfate during viral infection.

Isolation of a monoclonal antibody which blocks vaccinia virus infection.

We have isolated a monoclonal antibody, B2, that neutralizes vaccinia virus infection. B2 reacts with a trypsin-sensitive cell surface epitope. B2 does not neutralize infection of herpes simplex virus, suggesting that the B2-reactive epitope is specifically involved in vaccinia virus entry. A survey of 12 different cell lines reveals a correlation between B2 reactivity and susceptibility to vaccinia virus infection. In addition, B2 interferes with vaccinia virus adsorption to target cells. Taken together, the B2-reactive epitope is part of a receptor that appears important for vaccinia virus entry.

Strength and endurance in the therapeutic evaluation of prednisolone-treated MDX mice.

The dystrophin-deficient, X-linked dystrophic mouse (mdx) was used to evaluate the efficacy of prednisolone treatment. A test protocol was used to take advantage of the quantifiable weakness and disability as well as molecular genetic defect shared with the X-linked Duchenne muscular dystrophy (DMD). Whole-body weakness and fatigue were determined by non-invasive force-transducer physiographic and variable-speed treadmill techniques, respectively. Other measurements included hind-limb muscle protein, calcium, and histomorphology. Subcutaneously-administered prednisolone elicited significant improvements in whole body strength throughout a two-month test period. Increases in strength were also accompanied by measurable increments in running endurance. In fact, prednisolone treatment appeared to protect mdx mice from the stressful effect of continuous running as determined by strength and muscle fiber diameter. Test results from this study support the limited therapeutic benefit observed previously in DMD patients treated with the glucocorticoid.

Ectopic ACTH syndrome due to clear cell sarcoma of the kidney.

The 7-year-old girl presenting with cushingoid appearance and gross hematuria was found to have a right renal tumor. Preoperative hormonal assay revealed an extremely high plasma ACTH level and elevated plasma cortisol levels. Right radical nephrectomy was performed, and pathology proved the tumor to be a clear cell sarcoma, a sarcomatous variant of Wilms' tumor, of the kidney. Postoperative plasma ACTH level declined dramatically and cortisol levels returned to normal, too. She received chemotherapy and radiotherapy then and gradually lost her cushingoid appearance. This is the first reported case of clear cell sarcoma of the kidney with ectopic ACTH syndrome.