X-linked chronic granulomatous disease: correction of NADPH oxidase defect by retrovirus-mediated expression of gp91-phox

Chronic granulomatous disease (CGD) is an inherited immunodeficiency resulting from the inability of an individual's phagocytes to produce superoxide anions because of defective NADPH oxidase. The disease may be treated by bone marrow transplantation and as such is a candidate for somatic gene therapy. Two thirds of patients have defects in an X- linked gene (X-CGD) encoding gp91-phox, the large subunit of the membrane cytochrome b-245 component of NADPH oxidase. Epstein-Barr virus-transformed B-cell lines from patients with CGD provide a model system for the disease. We have used retrovirus-mediated expression of gp91-phox to reconstitute functionally NADPH oxidase activity in B-cell lines from three unrelated patients with X-CGD. The protein is glycosylated and membrane associated, and the reconstituted oxidase is appropriately activated via protein kinase C. The kinetics of superoxide production by such reconstituted cells is similar to that of normal B-cell lines. These data show the potential of gene therapy for this disease.     

New-Onset Diabetes and Preexisting Diabetes Are Associated With Comparable Reduction in Long-Term Survival After Liver Transplant: A Machine Learning Approach

Objective

To identify key predictors and survival outcomes of new-onset diabetes after transplant (NODAT) in liver transplant (LT) recipients by using the Scientific Registry of Transplant Recipients.

The isolation rate of Escherichia coli 0157:H7 in Toronto and surrounding communities

Verocytotoxin-producing strains of Escherichia coli, most often serotype 0157:H7, have been associated with both sporadic and epidemic diarrheal disease in Canada. In order to determine the isolation rate of E coli 0157:H7 in outpatients with diarrhea, all stool specimens submitted for culture to Med-Chem Laboratories in Metropolitan Toronto between June 1988 and September 1989 were cultured on MacConkey-Sorbitol agar in addition to standard enteric media. A total of 46 (0.3%) of 16,125 stool specimens yielded E coli 0157:H7 or verotoxin-producing E coli 0157:H(-). These isolates came from 31 patients with diarrhea; only 16 (52%) had a history of hemorrhagic colitis and one patient developed hemolytic uremic syndrome. Although MacConkey-Sorbitol agar was useful as a differential medium for detecting E coli 0157:H7, 14.5% of all specimens yielded nonsorbitol-fermenting isolates. It is not certain whether the routine use of MacConkey-Sorbitol agar is justified when isolation rates of E coli 0157:117 are very low.     

Blinded, placebo-controlled trial of antiparasitic drugs for trichinosis myositis

There is no consensus on the benefits of treatment with any specific anthelminthic compound on muscle-stage trichinosis. A double-blind, placebo-controlled comparison was done of 3 antiparasitic drugs during an outbreak of trichinosis in Chiangrai Province, northern Thailand. Forty-six adults were randomized to receive 10 days of oral treatment with mebendazole (200 mg twice a day), thiabendazole (25 mg/kg twice a day), fluconazole (400 mg initially, then 200 mg daily), or placebo. All patients received treatment to eradicate adult intestinal worms. Trichinella spiralis infection was proved parasitologically in 19 (41%) of 46 patient and by serodiagnosis in all cases. Significantly more patients improved after treatment with mebendazole (12/12) and thiabendazole (7/7) than after treatment with placebo (6/12; P<.05) or fluconazole (6/12). Muscle tenderness resolved in more patients treated with thiabendazole and mebendazole than in those treated with placebo (P<.05). However, 30% of volunteers could not tolerate the side effects of thiabendazole. In summary, Trichinella myositis responds to thiabendazole and to mebendazole.     

An improved negative immunomagnetic selection strategy for the purification of primitive hemopoietic cells from normal bone marrow

An improved negative immunomagnetic selection strategy has been devised for the enrichment of primitive hemopoietic cells using the high proliferative potential colony-forming cell (HPP-CFC) assay as an index of stem cell purification. Immunomagnetic selection was carried out using goat anti-rat conjugated M-450 Dynabeads and a cocktail of rat monoclonal antibodies directed against lineage antigens expressed on B-lymphocytes (B220), neutrophils and activated macrophages (7/4), differentiating erythroid cells (YW 25.12.7), and T-lymphocyte subsets (Lyt-2 and L3T4). This negative selection strategy results in the highly reproducible enrichment of HPP-CFC with negligible loss of HPP-CFC at the immunomagnetic selection step. A 30-fold enrichment of HPP-CFC stimulated by interleukin 3 (IL-3) plus colony-stimulating factor (CSF-1), or interleukin 1 alpha (IL-1 alpha) plus IL-3 plus CSF-1, is obtained with simultaneous resolution of HPP-CFC from progenitor cells of low proliferative potential responsive to CSF-1 alone (LPP-CFC). Flow cytometric analysis of these lineage-negative cells reveals that they almost exclusively exhibit the light-scattering characteristics of blast cells and the morphology of a candidate hemopoietic stem cell. Positive fluorescence-activated cell sorting of immunomagnetically pre-enriched normal bone marrow cells using wheat germ agglutinin yields cell preparations with a cloning efficiency of up to 45% and a HPP-CFC content of 20%.     

Detection of genomic deletions of PKP2 in arrhythmogenic right ventricular cardiomyopathy

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited myocardial disease that predominantly affects the right ventricle and is associated with ventricular arrhythmias that may lead to sudden cardiac death. Mutations within at least seven separate genes have been identified to cause ARVC, however a genetic culprit remains elusive in approximately 50% of cases. Although negative genetic testing may be secondary to pathogenic mutations within undiscovered genes, an alternative explanation may be the presence of large deletions or duplications involving known genes. These large copy number variants may not be detected with standard clinical genetic testing which is presently limited to direct DNA sequencing. We describe two cases of ARVC possessing large deletions involving plakophilin‐2 (PKP2) identified with microarray analysis and/or multiplex ligation‐dependent probe amplification (MLPA) that would have been classified as genotype negative with standard clinical genetic testing. A deletion of the entire coding region of PKP2 excluding exon 1 was identified in patient 1 and his son. In patient 2, MLPA analysis of PKP2 revealed deletion of the entire gene with subsequent microarray analysis demonstrating a de novo 7.9 Mb deletion of chromosome 12p12.1p11.1. These findings support screening for large copy number variants in clinically suspected ARVC cases without clear disease causing mutations following initial sequencing analysis.     

Dosimetric consequences of seed placement accuracy in permanent breast seed implant brachytherapy

PurposeThis study quantifies the dosimetric impact of implant accuracy and derives a quantitative relationship relating implant accuracy to target dosimetry.Methods and MaterialsA framework was developed to simulate multiple implants for error combinations. Spherical clinical target volumes (CTVs) were modeled with volumes 1.4 cm³, 9.2 cm³, and 20.6 cm³, representing the range seen clinically. Each CTV was expanded 10 mm isotropically to create a planning target volume (PTV).. Random and systematic seed placement errors were simulated by shifting needles from their planned positions. Implant errors were simulated over the range of clinically practical errors in permanent breast seed implant. The relative effect on target coverage was evaluated. Regression analysis was performed to derive relationships between CTV dosimetry and the magnitude of implant accuracy. The validity of the clinically used 10 mm PTV margin for each of the CTVs was assessed.ResultsIntroducing practical implant errors resulted in CTV V_90% median (10th and 90th percentile) of 97.7% (85.9% and 100%), 96.2% (86.8% and 99.7%), and 100% (77.8% and 100%) for the typical, large, and small CTV, respectively. All CTVs show similar trends in target coverage. Polynomials were derived relating seed placement accuracy to median (R² = 0.82) and 10th percentile (R² = 0.78) CTV V_90%._.ConclusionsThis work quantitatively describes the relationship between implant accuracy and CTV coverage. Based on simulations, the 10 mm PTV margin is adequate to maintain target coverage. These equations can be used with institutional seed placement accuracy to estimate coverage.     

Towards a consensus for classification of mandibular condyle fractures

The merits of surgical treatment of fractures of the mandibular condyle versus non-surgical management remains highly controversial, despite a large volume of literature dedicated to this topic. One reason the controversy remains, is because most of the outcomes in the literature are not directly comparable. The disparate range of condylar fracture classifications used is one of the reasons that studies are not comparable. We sought to review classification systems for condylar fractures used in the recent scientific literature.Review of the literature from 2016 to 2019, looking for papers relating to fractures of the mandibular condyle. Papers identified were assessed for type of study, focus of study, classification system used.88 studies were identified, including prospective and retrospective cohort studies, randomised and non-randomised prospective studies, randomised controlled trials and case series. More studies focussed on epidemiological factors than surgical access, fixation or outcomes. 31 used no classification system, whilst 17 used unique classification systems and 40 used previously referenced classification systems.Classification systems are used to help separate clinical problems into distinguishable groups, where there is a difference in management or outcome depending on the distinguishing features.There is currently a wide diversity of classification systems used for condyle fractures, and as a result, comparisons of surgical access, fixation and outcomes are difficult to make. Having a single classification system across the published literature would allow easier comparison and the classification proposed by the AO group is recommended for future use.