Clinical presentations and outcomes of Penicillium marneffei infections: a series from 1994 to 2004.

OBJECTIVES: To describe the clinical presentation, management, and outcomes of patients with Penicillium marneffei infections in a regional hospital in Hong Kong. DESIGN: Retrospective study. SETTING: A regional and tertiary human immunodeficiency virus-referral hospital in Hong Kong. PATIENTS: Those who had penicilliosis during the inclusive period January 1994 to February 2004. RESULTS: Forty-seven immunocompromised patients (44 being human immunodeficiency virus-positive) with penicilliosis were retrospectively studied. Fever, malaise, and anaemia were the commonest presentations. Most diagnoses were obtained from blood cultures (83%) and lymph node biopsies (34%). Five (11%) died, death being attributable to penicilliosis; four (9%) of them had received no specific antifungal treatment due to late presentation and late diagnosis. The CD4 count of human immunodeficiency virus-infected patients upon diagnosis of penicilliosis was low (median, 20.0 cells/mm3). Most (70%) patients received amphotericin B as an induction treatment, followed by oral itraconazole, although a smaller proportion (21%) received oral itraconazole only. All surviving human immunodeficiency virus-infected patients took highly active antiretroviral treatment and oral itraconazole as secondary prophylaxis after treatment of penicilliosis. The prognosis appeared satisfactory with early diagnosis and administration of appropriate antifungal therapy. Relapse ensued in two (4%) of the patients only. CONCLUSION: Penicillium marneffei infection in immunocompromised patients is a serious disease with significant mortality if not diagnosed early and treated with appropriate antifungal drugs. Simple investigations like blood culture enable the diagnosis in the majority of cases. Immunocompromised patients who have been successfully treated should receive oral itraconazole as a maintenance therapy to prevent relapse.     

Covalent linking of proteins and cytokines to suture: enhancing the immune response of head and neck cancer patients

BACKGROUND: The immune system of advanced stage head and neck cancer patients is frequently suppressed. Poor immune function has been correlated with poor clinical outcome. Immunotherapeutic strategies have been previously attempted in an effort to enhance immune function and improve survival. Previous studies have shown surgical suture can be transformed into an immune stimulant capable of activating the T lymphocytes of cancer patients. The development of a process for covalently linking proteins and cytokines to suture could have enormous potential for the in vivo manipulation of the immune system. HYPOTHESIS: We hypothesize proteins and cytokines can be covalently linked to surgical suture while preserving their functional properties. STUDY DESIGN: Prospective study testing normal donor and head and neck squamous cell carcinoma (HNSCC) patient lymphocytes. METHOD: Polyester suture was acid hydrolyzed followed by reacting with 1-ethyl-3(-3-dimethylamino propyl carbodiimide) (EDAC) to create a suture-EDAC intermediate. Next, selected proteins (horseradish peroxidase [HRP] or bovine serum albumin [BSA]) or cytokines (interleukin [IL]-2 or interferon [IFN]-gamma) were reacted with the suture-EDAC intermediate to test the covalent linkage of the selected protein or cytokine to suture. Functional activity of the linked proteins was measured spectrophotometrically. The linking of cytokines to suture was tested by stimulating normal donor peripheral blood lymphocytes (PBL) or HNSCC patients' lymphocytes. The functional activity was confirmed by proliferation, enzyme linked immunoadsorbent assay (ELISA), and phenotype expression of T cells.RESULTS The conditions for optimally linking a protein to polyester suture were defined using HRP as a model protein. HRP retained its enzymatic activity. The optimal conditions for linking IL-2 or IFN-gamma were defined. The covalently linked cytokines retained their immune enhancing properties for stimulating PBL and lymph node lymphocytes (LNL) from HNSCC patients to proliferate, generate a TH1 immunologic profile of cytokines (IL-2, IL-12, IFN-gamma), and stimulate T lymphocytes. CONCLUSION: This is the first report to demonstrate that cytokines can be covalently linked to surgical sutures and retain their immune-stimulating properties. Proteins linked to suture also retained their enzymatic activity. The clinical implications of functionally active cytokines or proteins linked to surgical suture may be very significant in the future for manipulating the immune system in vivo or enhancing wound healing.     

Collagen gel contraction by ARPE-19 cells is mediated by a FAK-Src dependent pathway

Proliferative vitreoretinopathy (PVR) may result in part from de-differentiation of retinal pigment epithelium (RPE) in an aberrant wound-healing strategy. An in vitro model of PVR, collagen gel contraction by RPE, likely requires integrin engagement and activation as an important initial step. The purpose of this study was to identify the important associated integrins and signal transduction pathway. The retinal pigment epithelial cell line ARPE-19 was used in these studies. Cell surface integrin expression was assessed using flow cytometry. An in vitro contraction assay was performed and the percent contraction quantified at specific time intervals using image capture (Gel Doc) and NIH Image software. Cells were pretreated with either small molecule inhibitors of signal transduction pathways or monoclonal antibodies with specificity for specific integrin isoforms. Transient transfections with a FAK siRNA were used to decrease FAK expression. ARPE-19 cells express alpha1, alpha2, and alpha3 integrin, isoforms involved in collagen ligation. Cell surface integrin blockade using anti-integrin alpha2 (P=0.02), alpha3 (P=0.01), or a combination of alpha1, alpha2, and alpha3 (P=0.001) antibodies significantly reduced collagen gel contraction. Inhibition of the FAK-Src complex, but not MEK or PI3K, significantly decreased contraction (P=0.0001). FAK siRNA transient transfection significantly reduced FAK protein expression by 71% (P=0.02) and concordantly decreased gel contraction (P=0.0001). RPE-mediated collagen gel contraction is a multi-step process. Integrin ligation and FAK-Src activation is necessary for collagen gel contraction produced by the ARPE-19 cell line. Validation of these observations in primary RPE cells may suggest new targets for therapeutic intervention in PVR.     

Inhibition of corneal angiogenesis by ascorbic acid in the rat model

Purpose To evaluate the effect of topically administered ascorbic acid on experimentally induced corneal neovascularization in the rat model. Materials and methods Corneal chemical cauterization of 72 eyes in Long-Evans male rats was performed using silver nitrate/potassium nitrate sticks. Nine groups of eight eyes were used to evaluate eight concentrations of ascorbic acid with one group of eight eyes serving as a control. Topical instillation of 100 mg/ml non-pH-neutralized ascorbic acid was performed in one group while the remaining seven groups were evaluated using pH-neutralized ascorbic acid in concentrations of 100 mg/ml, 50 mg/ml, 10 mg/ml, 5 mg/ml, 1 mg/ml, 500 μg/ml, and 250 μg/ml. Results The percentage of corneal neovascularization and burn stimulus score was determined for all the eyes. The means of percent of corneal neovascularization in ascorbic acid 100 mg/ml (non-neutralized), 100 mg/ml, 50 mg/ml, 10 mg/ml, 5 mg/ml, 1 mg/ml, 500 μg/ml, 250 μg/ml, and control group were 17.50 ± 12.80 (p = 0.001), 17.00 ± 19.30 (p = 0.001), 15.25 ± 13.26 (p = 0.001), 17.62 ± 11.89 (p = 0.001), 28.87 ± 23.08 (p = 0.001), 29.62 ± 16.91 (p = 0.001), 60.12 ± 8.50 (p = 0.04), 65.62 ± 2.26 (p = 0.185), and 68.25 ± 4.06, respectively (Tables 1 and 2). All animals had a burn score of 2+ or higher (Table 1). Conclusion Ascorbic acid applied in a topical solution appears to inhibit corneal neovascularization in the rat model of inflammatory neovascularization in concentrations in a dose-dependent manner. The optimal dose-effect relation was in our model found in concentrations between 1 mg and 500 μg/ml. At concentrations below 500 μg/ml there was no statistically significant inhibition in the degree of corneal neovascularization compared to control. None of the authors has any proprietary interest in any technique or product described herein.     

Peritoneal and intestinal tuberculosis: an ancestral disease that poses new challenges in the technological era. Case report and review of the literature

Tuberculosis is a public health problem. The most common presentation is pulmonary disease. The diagnosis of any extrapulmonary forms are quite difficult. Clinical manifestations of gastrointestinal tuberculosis are non-specific and compatible with pathologies such as inflammatory bowel disease, advanced ovarian cancer, deep mycosis, yersinia infection and amebomas. Abdominal form is located at 6th place of the extrapulmonary forms, after lymphatic, genitourinary, osteoarticular, miliary and meningeal infections. Eventually, 25 to 75% of patients with abdominal tuberculosis will require surgery. These procedures should be limitated with the purpose to preserve small bowel. Resection should be limitated for complicated cases. The surgical indications include: Intestinal occlusion (15-60%), perforation (1-15%), abscesses and fistulas (2-30%) and hemorrhage (2%). CONCLUSIONS: In most of the cases, the diagnosis of peritoneal or intestinal tuberculosis is made during a laparoscopy or laparotomy even during surgery performed by different purposes. Excessive manipulation of the intraabdominal organs may produced unexpected bowel lesions, increasing morbidity and mortality. Medical treatment is highly effective in the resolution of moderate complications such as bowel obstruction. Resectional procedures should be reserved for complications like perforation, bleeding or stenosis non-suitable for stricturoplasty.     

Urinary tract infection due to Corynebacterium urealyticum in kidney transplant recipients: an underdiagnosed etiology for obstructive uropathy and graft dysfunction-results of a prospective cohort study.

BACKGROUND: Corynebacterium urealyticum is a cause of urinary tract infection and encrusting cystitis or pyelitis. Information about this infection in renal transplant recipients is based on case reports. We communicate the first prospective epidemiological study for this population. METHODS: We selected a cohort of 163 renal transplant recipients who were screened for urinary tract infection due to C. urealyticum. Long-term incubation and special media were used for culture of C. urealyticum. The cohort was observed for a mean of 26.2 months (standard deviation, 8.7; range, 1-36 months). Risk factors and outcomes were assessed. RESULTS: At baseline, 16 (9.8%) of 163 patients had C. urealyticum bacteriuria (6 were asymptomatic, 9 had acute cystitis, and 1 had encrusting pyelitis). Independent risk factors (assessed by multivariate analysis) for urinary tract C. urealyticum infection were: antibiotic administration during the previous month (odds ratio, 8.04; 95% confidence interval, 1.57-41.06; P = .012), history of nephrostomy (odds ratio, 51.59; 95% confidence interval, 3.62-736.06; P = .004), and skin colonization (odds ratio, 208.35; 95% confidence interval, 21.54-2015.22; P< .001). Presence of urinary tract infection symptoms for >1 month (odds ratio, 27.7; 95% confidence interval, 2.55-300.5; P = .006) and obstructive uropathy (odds ratio 25.9; 95% confidence interval, 4.43-152.31; P < .001) were more frequent during follow-up in patients with C. urealyticum bacteriuria. CONCLUSIONS: When specifically tested for, C. urealyticum bacteriuria is more prevalent than previously thought in renal transplant recipients, and it is closely related to obstructive uropathy. Future studies are necessary to establish the relevance of treating the infection during follow-up after renal transplantation.