Routine histology techniques for the developing and adult central nervous system.

The preparation of tissue for histological study is a multi-step process in which potential loss of quality and the introduction of artifacts can occur during each step. Knowledge of the process and the potential pitfalls at each step will serve the investigator well. Here I describe the most basic histologic techniques, worked out so long ago they are often absent from current literature.

In vivo dual luciferase reporter assay with chick neural tube in ovo electroporation system.

Luciferase reporter systems are widely employed to provide a quantitative readout of gene expression for studies of transcriptional regulation, translation efficiency, and cell signaling. The most common application of luciferase involves transient transfections into cells in vitro or in vivo. In both cases, the normal variability inherent in transfection approaches can introduce significant errors into the data that makes comparison between separate experiments problematic.

Immunofluorescence staining with frozen mouse or chick embryonic tissue sections.

Immunofluorescence (IF), a form of immunohistochemistry (IHC) with specific applications, is commonly used for both basic research and clinical studies, including diagnostics, and involves visualizing the cellular distribution of target molecules (e.g., proteins, DNA, and small molecules) using a microscope capable of exciting and detecting fluorochrome compounds that emit light at specific, largely nonoverlapping wavelengths.

In ovo electroporation in embryonic chick spinal cords.

The developing spinal cord is a well-established model system widely used to study the signaling pathways and genetic programs that control neuronal/glial differentiation and neural circuit assembly. This is largely due to the relatively simple organization (compared to other CNS regions) and experimental accessibility of the neural tube, particularly in the chick embryo. In vivo transfection of cells within the developing chick neural tube using in ovo electroporation has emerged as a rapid and powerful experimental technique in that (1) transfected factors can be functionally tested in a spatially and temporally controlled manner and (2) the chick embryo provides a physiologically relevant in vivo environment to conduct biochemical studies such as dual-channel luciferase assay, co-immunoprecipitation (co-IP), and Chromatin Immunoprecipitation (ChIP).

Culturing oligodendrocyte lineage cells from neonatal rats.

The use of enriched oligodendrocyte lineage cell cultures has yielded insight into functions of these cells and regulatory mechanisms. This chapter details methods that result in such cultures.

Culturing astrocytes from postnatal rats.

The use of cultures has informed us of functions and regulation of astrocytes that were previously unknown. This chapter details the methods that result in such cultures.

Nucleoside analog labeling of neural stem cells and their progeny.

Nucleoside analog pulse labeling is an important technique which can assess the birthdate, cell cycle maintenance, or cycling rates of cells during development. This method has evolved over several decades of use and is now applied to a multitude of tissue subtypes and systems. The methodology in this chapter covers the classic uses for analog pulse labeling as well as their use in conjunction with the newly characterized technique of in utero electroporation (IUE).

Patients with unstable slipped capital femoral epiphysis have antecedent symptoms.

Background: The characteristics of patients who sustain unstable slipped capital femoral epiphyses (SCFEs) are not well described compared to their counterparts who sustain stable SCFE. Although patients with unstable slips are usually identified owing to acute symptoms, it is unclear whether these patients have premonitory symptoms that could heighten the awareness of treating physicians to the possibility of an unstable slip and lead to timely diagnosis and treatment.

Questions/purpose: We determined whether most patients experienced pain and limp before developing an unstable SCFE.

Telangiectasia macularis eruptiva persians presenting as island sparing.

Mastocytosis is characterized by the proliferation and accumulation of mast cells within organs and most commonly the skin; localization accounting for the frequent presentation of skin lesions in affected individuals. The authors detail a case report involving a patient with telangiectasia macularis eruptive perstans, a rare cutaneous form of mastocytosis, accompanied by an unusual clinical finding of island sparing.

Cancer Survivors and the Patient-Centered Medical Home.

Survivor care plans have been described as useful tools for enhancing the quality of follow-up care that cancer survivors receive after their active treatment has been completed. The relative success of current survivor care plan models is strongly dependent on the actions of individual patients. In this qualitative study of 33 cancer survivors, we explored patients' understanding of follow-up care and their motivations and resources for seeking care.

Bone morphogenetic protein type I receptor antagonists decrease growth and induce cell death of lung cancer cell lines.

Bone morphogenetic proteins (BMPs) are highly conserved morphogens that are essential for normal development. BMP-2 is highly expressed in the majority of non-small cell lung carcinomas (NSCLC) but not in normal lung tissue or benign lung tumors. The effects of the BMP signaling cascade on the growth and survival of cancer cells is poorly understood.

The Notch signaling pathway as a mediator of tumor survival.

The Notch signaling pathway is evolutionarily conserved and responsible for cell fate determination in the developing embryo and mature tissue. At the molecular level, ligand binding activates Notch signaling by liberating the Notch intracellular domain, which then translocates into the nucleus and activates gene transcription. Despite the elegant simplicity of this pathway, which lacks secondary messengers or a signaling cascade, Notch regulates gene expression in a highly context- and cell-type-dependent manner.

Lumbar hemangioma masking a plasma cell tumor--case report and review of the literature.

Background Context: Vertebral hemangiomata are ubiquitous bone tumors. Often multiple, they are generally benign in nature and slow growing. They typically have a predictable radiographic appearance.

Amyopathic dermatomyositis complicated by pneumomediastinum.

Dermatomyositis is an inflammatory disease of unclear etiology with characteristic cutaneous and musculoskeletal findings. Amyopathic dermatomyositis is a subtype without musculoskeletal involvement. Many cases of dermatomyositis are associated with underlying malignancy, but pulmonary manifestations can also be seen, the most common of which is interstitial lung disease.

Evolution of Sindbis virus with a low-methionine-resistant phenotype is dependent both on a pre-existing mutation and on the methionine concentration in the medium.

SVlm21 is a mutant of Sindbis virus which was isolated by serial passage of virus in mosquito cells maintained in low-methionine medium; it therefore has a low-methionine-resistant (LMR) phenotype. This phenotype requires mutations at nts 319 and 321; these mutations result in Arg to Leu and Ser to Cys changes at positions 87 and 88 respectively in the viral methyl transferase, nsP1. To better understand the genesis of SVlm21, we carried out serial passages of viruses having only one of these amino acid changes, but in mosquito cells maintained in normal methionine-medium.

Pharmacist-led workshops to enhance pharmacotherapy knowledge for medical students.

Background: Graduating medical students have reported concern regarding inadequate training in pharmacotherapy. Teaching by clinical pharmacists may improve medical students' pharmacotherapy knowledge.

Purpose: To assess the impact of pharmacist led workshops on 4th year medical students' knowledge of pharmacotherapy and satisfaction.

Interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) as a prognostic marker for local control in T1-2 N0 breast cancer treated with breast-conserving surgery and radiation therapy (BCS + RT).

Interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) expression, involved in the regulation of translation, has been implicated to mediate resistance to chemotherapy and radiation in cancer cells in vitro. The purpose of this study was to evaluate the prognostic significance of IFIT1 protein expression in patients with breast cancer treated with Breast-Conserving Surgery and Radiation Therapy (BCS + RT). A tissue microarray was constructed with specimens from 282 women with node-negative, early-stage (I/II) breast cancer who were treated with BCS + RT.

Stimulus Sensitive Constant Micro Macro Square-Wave Jerks in a Comatose Patient.

Saccades are generally thought of as being cortically generated and not seen in comatose patients. We describe a patient, comatose secondary to a large intracerebral hemorrhage, who developed constant rhythmic small amplitude square-wave jerks with an intersaccadic interval of 130 ms. Despite the patient's comatose state the eye movements would stop transiently in response to auditory or tactile stimulation and then quickly regain their previous amplitude.

WAVE/SCAR promotes endocytosis and early endosome morphology in polarized C. elegans epithelia.

Cells can use the force of actin polymerization to drive intracellular transport, but the role of actin in endocytosis is not clear. Studies in single-celled yeast demonstrate the essential role of the branched actin nucleator, Arp2/3, and its activating nucleation promoting factors (NPFs) in the process of invagination from the cell surface through endocytosis. However, some mammalian studies have disputed the need for F-actin and Arp2/3 in Clathrin-Mediated Endocytosis (CME) in multicellular organisms.

Update on blood cultures: how to obtain, process, report, and interpret.

The detection and identification of microorganisms circulating in the bloodstream of patients is arguably one of the most important functions of the clinical microbiology laboratory. Effective implementation of this function requires careful consideration of specimen collection and processing, culture techniques, result reporting, and, perhaps most importantly, result interpretation by the physician. The purpose of this review is to provide a synopsis of the current state of the art for each of these areas, with the intention of providing adequate information to enable clinical laboratory personnel and physicians to critically evaluate and, if required, improve their current blood culture practices.

Effect of interferon beta-1a on B7.1 and B7.2 B-cell expression and its impact on T-cell proliferation.

The effect of pharmacologically relevant doses of interferon (IFN) β-1a on B-cell expression of B7.1 and B7.2 was investigated.

Floor plate-derived sonic hedgehog regulates glial and ependymal cell fates in the developing spinal cord.

Cell fate specification in the CNS is controlled by the secreted morphogen sonic hedgehog (Shh). At spinal cord levels, Shh produced by both the notochord and floor plate (FP) diffuses dorsally to organize patterned gene expression in dividing neural and glial progenitors. Despite the fact that two discrete sources of Shh are involved in this process, the individual contribution of the FP, the only intrinsic source of Shh throughout both neurogenesis and gliogenesis, has not been clearly defined.