Development and evaluation of a quantitative, touch-down, real-time PCR assay for diagnosing Pneumocystis carinii pneumonia

A rapid (time to completion, <4 h, including DNA extraction) and quantitative touch-down (QTD) real-time diagnostic Pneumocystis carinii PCR assay with an associated internal control was developed, using fluorescence resonance energy transfer (FRET) probes for detection. The touch-down procedure significantly increased the sensitivity of the assay compared to a non-touch-down procedure. Tenfold serial dilutions of a cloned target were used as standards for quantification. P. carinii DNA has been detected in respiratory specimens from patients with P. carinii pneumonia (PCP) and from patients without clinical evidence of PCP. The latter probably represents colonization or subclinical infection. It is logical to hypothesize that quantification might prove helpful in distinguishing between infected and colonized patients: the latter group would have lower copy numbers than PCP patients. A blinded retrospective study of 98 respiratory samples (49 lower respiratory tract specimens and 49 oral washes), from 51 patients with 24 episodes of PCP and 34 episodes of other respiratory disease, was conducted. PCR-positive samples from colonized patients contained a lower concentration of P. carinii DNA than samples from PCP patients: lower respiratory tract samples from PCP and non-PCP patients contained a median of 938 (range, 2.4 to 1,040,000) and 2.6 (range, 0.3 to 248) (P < 0.0004) copies per tube, respectively. Oral washes from PCP and non-PCP patients contained a median of 49 (range, 2.1 to 2,595) and 6.5 (range, 2.2 to 10) (P < 0.03) copies per tube, respectively. These data suggest that this QTD PCR assay can be used to determine if P. carinii is present in respiratory samples and to distinguish between colonization and infection.     

Episcleral pseudomelanoma: late complication of scleral tunnel incision

PURPOSE: To describe two patients who developed scleral thinning as a late complication of scleral tunnel incision for cataract extraction. DESIGN: Observational case report and interventional case report. METHODS: A 75-year-old man had an asymptomatic gray-brown subconjunctival nodule in the location of the healed scleral tunnel incision for cataract surgery. A 62-year-old man had a biopsy of a brown conjunctival nodule at the site of a scleral tunnel incision for cataract surgery. RESULTS: The 75-year-old patient with prolapsed uveal tissue was followed up for 4 years without change in the wound defect. The 62-year-old patient had a biopsy because the exposed ciliary body mimicked a melanoma. CONCLUSIONS: Scleral defects with herniated uveal tissue as a late sequela of scleral tunnel incision for cataract surgery should be differentiated from melanoma and can be safely observed for years when covered by conjunctiva.     

Interdisciplinary rounds: impact on patients,families, and staff

Interdisciplinary rounds are a new care coordination strategy in several healthcare settings. This article describes the process used by clinical nurse specialists in one institution to broaden existing discharge planning rounds to interdisciplinary rounds. In addition, a survey queried advanced practice nurse subscribers to two listserves, the ANPACC and CNS-L, to determine how other institutions conducted interdisciplinary rounds, including structure, membership, and leadership. As a result of the changed focus in rounds, the target institution noted greater participation by all disciplines in achieving patient and family outcomes, increased early recognition of patients at risk, and improved communication among members of the healthcare team. These findings were supported by participants in the Internet survey. It was determined that interdisciplinary rounds are a valuable mechanism for improved patient outcomes and increased staff professionalism.     

Anatomic and histopathologic findings following a failed ahmed glaucoma valve device

Ahmed glaucoma valve implant (AGV) is one of the more commonly used implants in difficult glaucomas. The histology of a functioning bleb following AGV implantation and its anatomic relationship to the optic nerve when placed in the superionasal quadrant has been described. We report the histology of a failed bleb and the anatomic relationship between the optic nerve and the AGV end-plate when placed 9 mm from the limbus in a patient with neovascular glaucoma.     

Detection of viral-specific nucleic acid and intracellular virions in ventral horn neurons of lactate dehydrogenase-elevating virus infected C58 mice

C58 mice which have been immunosuppressed by treatment with cyclophosphamide (200 mg/kg) one day prior to infection with the C strain of lactate dehydrogenase-elevating virus (LDV-C) develop poliomyelitis. Using in situ hybridisation, we found that some ventral horn neurons in these mice contain cytoplasmic viral-specific nucleic acid. Viral-specific nucleic acid was also found within a few small cells located near inflammatory foci. In addition, mature virus particles were observed by electron microscopy in some ventral horn neurons, indicating that these cells are productively infected in C58 mice. Neither viral nucleic acid nor virions were found in the ventral horn neurons of poliomyelitis-resistant mouse strains or C58 mice that were not immunosuppressed prior to infection. Ventral horn neurons which contained viral nucleic acid or virions within cytoplasmic vesicles generally were normal in appearance and were not located within poliomyelitis inflammatory foci. Our data are consistent with the hypothesis that infected neurons first replicate virus and subsequently are attacked and cleared by inflammatory cells.