Site-Specific Molecular Diffusion in Articular Cartilage Measured using Fluorescence Recovery after Photobleaching

Diffusive transport of solutes is critical to the normal function of articular cartilage. The diffusion of macromolecules through cartilage may be affected by the local composition and structure, which vary with depth from the tissue surface. We hypothesized that the diffusion coefficient of uncharged molecules also varies with depth and molecular size. We used fluorescence recovery after photobleaching (FRAP) to measure site-specific diffusion coefficients of fluorescent dextran molecules (3, 40, 70, and 500 kDa) in porcine articular cartilage. The diffusion coefficients measured using FRAP exhibited an inverse size dependence and were in general agreement with those measured using other techniques. The diffusion coefficients for all molecules varied significantly with depth in a manner that depended upon the size of the diffusing molecule. The diffusion coefficients for the 3 and 500 kDa dextrans were 1.6 and 2.4 times greater, respectively, in the surface zone as compared to the middle and deep zones, whereas the diffusion coefficients of the 40 and 70 kDa dextrans were 0.3 and 0.2 times lower in the surface zone as compared to the middle and deep zones. These differences may reflect variations in the structure and composition of collagen, proteoglycans, and other macromolecules among the zones.© 2003 Biomedical Engineering Society. PAC2003: 8715Vv, 8719Rr, 8764Tt     

Rapid analysis of trans,trans-muconic acid in urine using microextraction by packed sorbent

A simple and fast method has been developed for the determination of urinary trans,trans-muconic acid (ttMA) based on micro-extraction by packed sorbent (MEPS), followed by HPLC-UV detection. Parameters affecting the performance of MEPS were investigated. The results of the MEPS-HPLC-UV procedure were compared with the results of the conventional SPE method. The procedure was used for the determination of ttMA in urine samples of benzene-exposed subjects. The calibration curve was obtained in the range of 0.1 to 2 μg/mL, and the method showed high linearity (R²=0.9997). The LOQ was 0.1 μg/mL. For the MEPS method, the withinand between-day precision ranged from 3.0 to 5.1% and 3.3 to 5.2%, respectively. The within-and between-day accuracy ranged from 93.3 to 99.3% and 91.5 to 96.0%, respectively. The developed MEPS-HPLC-UV method is suggested as an alternative to the existing conventional SPE method for the biomonitoring of benzene-exposed subjects.     

Stakeholders’ Perceptions Regarding the Use of Patient Photographs Integrated with Medical Imaging Studies

Integrating digital facial photographs of pediatric patients as identifiers (ID) with medical imaging (integrated photographic IDs) may increase the detection of mislabeled studies. The purpose of this study was to determine how different stakeholders would receive this novel technology. Parents or guardians of patients in a children’s hospital outpatient radiology department, radiology faculty and residents, and radiology technologists and nurses were asked to complete a survey. The perception about the anticipated use of integrated photographic ID in different clinical scenarios was investigated, and its predictors were determined using logistic regression analysis. Four hundred ninety-eight parents responded (response rate 83 %); 96 and 97 % supported the use of integrated photographic ID, if it improves the radiologist’s imaging interpretation or decreases the rate of mislabeled errors, respectively. Thirty-eight percent were worried that photographic IDs would impact patients’ privacy. Ninety-four percent believed that they should be asked for their consent prior to obtaining their child’s photograph. Seventy-eight radiologists responded (response rate 39 %); 63 and 59 % believed that the use of integrated photographic ID would result in improvement in accurate interpretation of images and identification of mislabeled patient errors, respectively. Forty-nine percent of radiologists had concern that integrated photographic ID would increase interpretation time. Fifty technologists and nurses responded (response rate 59 %); 71 and 73 % supported the technology if it resulted in more acute interpretation of images and identification of mislabeled patients, respectively. A majority of stakeholders support integrated photographic ID in order to improve safety. A majority of parents believe that consent should be obtained.     

Comprehensive characterization of a Canadian cohort of von Hippel-Lindau disease patients

Von Hippel-Lindau disease (VHL) is a heritable condition caused by pathogenic variants in VHL and is characterized by benign and malignant lesions in the central nervous system (CNS) and abdominal viscera. Due to its variable expressivity, existing efforts to collate VHL patient data do not adequately capture all VHL manifestations. We developed a comprehensive and standardized VHL database in the web-based application, REDCap, that thoroughly captures all VHL manifestation data. As an initial trial, information from 86 VHL patients from the University Health Network/Hospital for Sick Children was populated into the database. Analysis of this cohort showed missense variants occurring with the greatest frequency, with all variants localizing to the α- or β-domains of VHL. The most prevalent manifestations were central nervous system (CNS), renal, and retinal neoplasms, which were associated with frameshift variants and large deletions. We observed greater age-related penetrance for CNS hemangioblastomas with truncating variants compared to missense, while the reverse was true for pheochromocytomas. We demonstrate the utility of a comprehensive VHL database, which supports the standardized collection of clinical and genetic data specific to this patient population. Importantly, we expect that its web-based design will facilitate broader international collaboration and lead to a better understanding of VHL.     

Effects of histidine and n-acetylcysteine on experimental lesions induced by doxorubicin in sciatic nerve of rats

In this study, the effect of separate and combined intraperitoneal (i.p.) injections of histidine and n-acetylcysteine were investigated on experimental damage induced by doxorubicin (DOX) in sciatic nerve of rats. DOX was i.p. injected at a dose of 4?mg/kg once weekly for four weeks. Histidine and n-acetylcysteine were i.p. injected at a same dose of 20?mg/kg. Cold and mechanical allodynia were recorded using acetone spray and von Frey filaments tests, respectively. The sciatic nerve damage was evaluated by light microscopy. Plasma levels of malondialdehyde (MDA) and total antioxidant capacity (TAC) were measured. Histidine and especially n-acetylcysteine at a same dose of 20?mg/kg suppressed cold and mechanical allodynia, improved sciatic nerve lesions and reversed MDA and TAC levels in DOX-treated groups. Combination treatment with histidine and n-acetylcysteine showed better responses when compared with them used alone. The results of the present study showed peripheral neuroprotective effects for histidine and n-acetylcysteine. Reduction of free radical-induced toxic effects may have a role in neuroprotective properties of histidine and n-acetylcysteine.     

Ten Years’ Experience in Surgical Treatment of Right Middle Lobe Syndrome

Purpose: In this study we present the clinical, radiological, pathological, bronchoscopic and surgical results of 40 patients with diagnosis of middle lobe syndrome who were referred to our thoracic surgery unit for surgical intervention in a 10 years period.Methods: Forty patients with obstructive and non-obstructive causes of middle lobe syndrome referred to our thoracic surgery unit. Clinical data were collected from the patients’ records in a ten years period. This study evaluates diagnostic approaches and surgical treatments in right middle lobe syndrome.Results: We studied 23 females (57.5%) and 17 males (42.5%) with a mean age of 31.7. Clinical findings were cough 95%, sputum 80% and intermittent hemoptysis in 50% of patients. Middle lobe collapse was seen in CT scan of all patients. Bronchiectasis was the most common pathologic finding (55%). Tuberculosis was not rare and was final pathology in 20% of patients. In three patients ruptured hydatid cyst was final finding. Surgery was done without mortality and with only minor complications.Conclusion: Lobectomy of right middle lobe is a good therapeutic option in these patients. Due to high prevalence of tuberculosis and hydatid cyst in Middle Eastern countries these two must be considered as causes of middle lobe syndrome.     

Mitotic Index is an Independent Predictor of Recurrence‐Free Survival in Meningioma

While World Health Organization (WHO) grading of meningioma stratifies patients according to recurrence risk overall, there is substantial within‐grade heterogeneity with respect to recurrence‐free survival (RFS). Most meningiomas are graded according to mitotic counts per unit area on hematoxylin and eosin sections, a method potentially confounded by tumor cellularity, as well as potential limitations of accurate mitotic figure detection on routine histology. To refine mitotic figure assessment, we evaluated 363 meningiomas with phospho‐histone H3 (Ser10) and determined the mitotic index (number of mitoses per 1000 tumor cells). The median mitotic indices among WHO grade I (n = 268), grade II (n = 84) and grade III (n = 11) tumors were 1, 4 and 12. Classification and regression tree analysis to categorize cut‐offs identified three subgroups defined by mitotic indices of 0–2, 3–4 and ≥5, which on univariate analysis were associated with RFS (P < 0.01). In multivariate analysis, mitotic index subgrouped in this manner was significantly associated with RFS (P < 0.01) after adjustment for Simpson grade, WHO grade and MIB‐1 index. Mitotic index was then examined within individual WHO grade, showing that for grade I and grade II meningiomas, mitotic index can add additional information to RFS risk. The results suggest that the use of a robust mitotic marker in meningioma could refine risk stratification.