Cell-to-Cell Transcription Variability as Measured by Single-Molecule RNA FISH to Detect Epigenetic State Switching.

Single-molecule RNA fluorescent in situ hybridization (smRNA FISH) allows for the visualization, localization, and quantification of RNA transcripts within individual cells and tissues using custom-designed fluorescently labeled oligonucleotide probes. Here we describe a protocol for the preparation, imaging, and analysis of a smRNA FISH experiment that can be applied to any RNA of choice. We also provide insights as to how this powerful tool can be used to study epigenetic regulation, for example, following the epigenetic editing of genes.

Surgical wound morbidity in an austere surgical environment.

Surgical wound morbidity was analyzed for a U.S. military field hospital deployed to the Republic of Haiti in support of Operation New Horizons 1998. The purpose of the analysis was to determine whether procedures performed in the field hospital had greater infectious risks as a result of the environment compared with historical reports for traditional hospital or clinic settings. Acceptable historical infection rates of 1.5% for clean surgical cases, 7.7% for clean contaminated cases, 15.2% for contaminated cases, and 40% for dirty cases have been noted. There were 827 operations performed during a 6-month period, with the majority of patients assigned American Society of Anesthesiologists (ASA) Physical Status Classification class I or II. The distribution of these cases was: 72% clean cases, 5% clean contaminated cases, 4% contaminated cases, and 19% dirty cases. The overall wound complication rate was 3.6%, which included 5 wound infections, 11 wound hematomas, 8 superficial wound separations, and 6 seromas. The infectious morbidity for clean cases, the index for evaluation of infectious complications, was 0.8%, well within the accepted standards. There were two major complications that required a return to the operating room: a wound dehiscence with infection in an orchiectomy and a postoperative hematoma with airway compromise in a subtotal thyroidectomy. There were no surgical mortalities. The infectious wound morbidity for operations performed in the field hospital environment was found to be equivalent to that described for the fixed hospital or clinic settings. No special precautions were necessary to ensure a low infection rate. The safety for patients undergoing elective surgical procedures has been established. Further training using these types of facilities should not be limited based on concerns for surgical wound morbidity.

Cerebellar herniation after cervical transforaminal epidural injection.

OBJECTIVE: The purpose of this study is to inform readers of potential catastrophic complications associated with performing cervical transforaminal epidural steroid injections. CASE REPORT: A 31-year-old man presented to the pain clinic with a history of cervical radicular pain and right upper-extremity radicular symptoms. He was referred from the orthopedic spine clinic for evaluation for epidural steroid injection, having failed conservative treatment consisting of nonsteroidal anti-inflammatory drugs, muscle relaxants, oral steroids, and physical therapy. After undergoing a transforaminal epidural steroid injection at the right C8 nerve root, he developed a cerebellar infarct and brainstem herniation. He survived but has residual deficits of persistent diplopia on right lateral gaze and difficulties with short-term memory loss and concentration. CONCLUSIONS: Although transforaminal epidural steroid injections are an efficacious treatment for radicular syndromes, there can be catastrophic complications. In light of the growing body of similar case reports, further investigation is warranted to establish a safe protocol for the use of this modality.