Polymerase Chain Reaction and Goldmann-Witmer Coefficient Testing in the Diagnosis of Infectious Uveitis in HIV-Positive and HIV-Negative Patients in South Africa

Purpose: To use polymerase chain reaction (PCR) and Goldmann-Witmer coefficient (GWC) calculation to diagnose infectious uveitis.

Methods: Prospective cross-sectional study.

Results: Twenty-seven of 106 patients had positive PCR and/or GWC results on aqueous humor (AH) sampling and 15 of 27 (55.6%) were HIV-positive. Patients with non-anterior uveitis (NAU) were more likely to be HIV+ (p = 0.005). More than 1 possible pathogen was identified in 9 of 27 patients of whom 7 were HIV+. The final clinical diagnosis was discordant with AH findings in 9 of 27 cases. A positive EBV PCR result was associated with a discordant diagnosis (p = 0.001). All cases of herpetic anterior uveitis (42.9% HIV+) tested PCR-/GWC+ while all cases of herpetic NAU tested PCR+/GWC- (83.3% HIV+). All rubella virus cases were PCR+/GWC+.

Conclusion: PCR is useful to diagnose herpetic NAU in HIV+ patients while GWC is useful to diagnose herpetic anterior uveitis.     

Immediate postgadolinium spoiled gradient-echo MRI for evaluating the abdominal aorta in the setting of abdominal MR examination

To assess the reproducibility and image quality of immediate postgadolinium chelate spoiled gradient-echo MRI in demonstrating disease of the abdominal aorta. All patients (27 patients: 21 men, 6 women) with substantial disease of the abdominal aorta, who underwent abdominal MR examinations at 1.5 T between 1991 and 1995, were entered in the study. Patients were referred for evaluation of suspected aortic disease (14 patients) or other abdominal diseases (13 patients). Three experienced investigators manually measured luminal and external aortic wall diameters and rated image quality, definition of inner and outer walls, extent of disease, and presence of other abdominal abnormalities, in an independent fashion. A cardiovascular surgeon then rated all studies to determine whether clinical management could be based on the MR findings alone. There was 98 to 99% agreement in measurements of luminal and external wall diameter between the three investigators. Overall image quality was rated as good in 77.8 to 88.9% of patients. A total of 31 additional nonaortic abdominal abnormalities were detected by all observers. The cardiovascular surgeon rated 25 of 27 studies as adequate to determine clinical management based on MR findings alone. Immediate postgadolinium spoiled gradient-echo MRI is a reproducible technique for the demonstration of abdominal aortic disease and possesses good image quality. Advantages of this technique include simultaneous evaluation of other nonvascular diseases of the abdomen, short examination time, and easy implementation as part of routine abdominal MRI scanning protocol.     

Vitamin C-induced loss of redox-dependent viability in lung microvascular endothelial cells

Recent clinical trials have shown that vitamin C, at pharmacological concentrations (milligram to approximately gram), upon infusion into circulation, modulates vasodilation and vascular tone in humans. This also results in the elevated concentrations of vitamin C in circulation in the millimolar range. Here, it was hypothesized that vitamin C at pharmacological concentrations (millimolar) would induce oxidative stress and cause loss of redox-dependent cell viability in vascular endothelial cells (ECs). To test the hypothesis, bovine lung microvascular ECs (BLMVECs) in monolayer cultures were exposed to vitamin C (0-10 mM) for different time periods (0-2 h). Electron paramagnetic resonance spectroscopy revealed the intracellular formation of ascorbate free radical in a dose- and time-dependent fashion. Vitamin C also induced formation of intracellular reactive oxygen species in a dose-dependent fashion. It was observed that vitamin C induced morphological alterations and loss of cell viability in a dose- and time-dependent fashion, as measured by light microscopy and Alamar Blue redox cell viability assay, respectively. Vitamin C analogues failed to induce such changes. Vitamin C depleted cellular GSH levels in a dose-dependent fashion, suggesting that vitamin C altered thiol-redox status in BLMVECs. Antioxidants, intracellular iron chelator, and catalase protected cells against vitamin C-induced loss of redox-dependent cell viability, confirming the role of hydrogen peroxide and iron during redox cycling of vitamin C. These results, for the first time in detail, established that vitamin C at pharmacological doses induced oxidative stress and loss of redox-dependent cell viability in microvascular ECs.     

DNA end-independent activation of DNA-PK mediated via association with the DNA-binding protein C1D

DNA-dependent protein kinase (DNA-PK), which is involved in DNA double-strand break repair and V(D)J recombination, is comprised of a DNA-targeting component termed Ku and an ~465-kD catalytic subunit, DNA-PK_cs. Although DNA-PK phosphorylates proteins in the presence of DSBs or other discontinuities in the DNA double helix in vitro, the possibility exists that it is also activated in other circumstances via its association with additional proteins. Here, through use of the yeast two-hybrid screen, we discover that the recently identified high affinity DNA binding protein C1D interacts with the putative leucine zipper region of DNA-PK_cs. Furthermore, we show that C1D can interact with DNA-PK in mammalian cells and that C1D is a very effective DNA-PK substrate in vitro. Finally, we establish that C1D directs the activation of DNA-PK in a manner that does not require DNA termini. Therefore, these studies provide a function for C1D and suggest novel mechanisms for DNA-PK activation in vivo.     

Isolated cardiac recurrence of acute lymphoblastic leukemia characterized by t(11;19) two years after unrelated allogeneic bone marrow transplantation

A 33-year-old male presented with acute lymphoblastic leukemia (ALL) characterized by translocation (11;19)(q23;p13.3). He received an allogeneic bone marrow transplant from a matched unrelated donor. Two years later his disease relapsed with an isolated intracardiac mass, presenting as right heart failure. He had no evidence of concomitant relapse in the bone marrow. Tumor cytogenetics revealed clonal evolution with the karyotype 46,XY,t(3;16)(q23;p13),t(11;19)(q23;p13.3), the chromosome 16 breakpoint involving the band where the genes for multidrug resistance-associated protein and CREB binding protein are known to reside. To our knowledge, this is the first report of an isolated extramedullary relapse of ALL in the heart.     

Constitutive activation of the PI3K‐AKT pathway and cardiovascular abnormalities in an individual with Kosaki overgrowth syndrome

Kosaki overgrowth syndrome is a recently described syndrome characterized by distinctive facial features, brain white matter lesions, and developmental delay. Germline activating heterozygous PDGFRB mutations have been reported in this condition. Systemic connective tissue‐type findings have been described in some individuals. We describe a 19‐year‐old Caucasian female with a history of hydrocephalus, Dandy–Walker malformation, cervical spine arachnoid cyst, progressive scoliosis, and overgrowth. Her physical exam included distinctive craniofacial dysmorphism, as well as soft and hyperextensible skin. Cardiovascular imaging during adolescence revealed saccular aneurysms in both coronary artery systems and subtle tortuosity of the cervical vertebral arteries. Exome sequencing trio analysis identified a de novo previously reported pathogenic variant in PDGFRB, c.1696T>C (p.[Trp566Arg]). Further functional studies included platelet‐derived growth factor cellular metabolic pathway activity that confirmed the variant causes a constitutive activation of the PI3K‐AKT pathway. This is the first report to characterize the activating nature of this PDGFRB variant. We also highlight the connective tissue findings seen in Kosaki overgrowth syndrome and recommend baseline echocardiographic evaluation in all individuals with this condition with particular emphasis on coronary arteries.     

Health-related quality of life and sleep-disordered breathing in children

STUDY OBJECTIVES: To assess the extent to which sleep-disordered breathing (SDB) is associated with impairment of health-related quality of life (HRQOL) in children DESIGN: Observational study of pediatric participants in a longitudinal genetic-epidemiologic cohort study of SDB. SETTING: Community-based; studies conducted at participants' homes PARTICIPANTS: 298 children, aged 11.1 years +/- 3.5 SD; 54% females; 61% African-American or Other; 81% with a family member identified with laboratory-confirmed SDB. INTERVENTIONS: Not applicable MEASUREMENTS AND RESULTS: The HRQOL was assessed with the Child Health Questionnaire (CHQ-PF50), a 50-item parent-completed form that measures 14 multidimensional health concepts. Sleep-disordered breathing (SDB) was assessed with ovemight in-home monitoring that recorded nasal-oral airflow, pulse oximetry, chest-wall impedance, and heart rate. Using logistic regression analyses, each CHQ scale outcome was analyzed independently for the effect of SDB, adjusting for other potential confounding variables and for family-correlated data. Significant differences in overall physical health and complaints of bodily pain were observed in children with generally mild levels of SDB. Relationships persisted after adjustment for age, gender, ethnicity, obesity, and asthma. CONCLUSIONS: SDB in children is associated with measurably lower levels of specific dimensions of HRQOL in children. Decrements in HRQOL are measurable even for children with mild SDB, with increasing effects observed with more severe SDB.