An effect of prostaglandin E1 on the acinar cell of the rat parotid gland

The effects of in vivo administration of prostaglandin E1 (PGE1) on rat parotid gland acinar cells were studied and compared with glands removed from animals which had been either fasted or stimulated to discharge stored secretory granules by an injection of isoproterenol (IPR). Depletion of tissue amylase, increased plasma amylase, and alterations in secretory cell ultrastructure were used to assess the effects of PGE1. Initially following PGE1 administration, the secretory granules enlarged their matrices and lost electron opacity, and numerous fusions between secretory granules occurred. Concomitant with the above events, material having an appearance identical to that within the secretory granules was observed in the acinar lumina and the intercellular spaces. The PGE1-induced release of stored secretory materials from storage granules was transient. Five hours after the onset of hourly PGE1 injections, the biochemical and structural features of the gland were "normal." The effects of PGE1 on the rat parotid gland are discussed. A theory of PGE1 action based on the translocation of ionic calcium is proposed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/22258/1/0000695.pd

Electron microscopy of cytoplasmic crystalloids in rat parotid glands

Crystalloid-containing bodies that may be lysosomes were observed by electron microscopy in acinar and striated duct cells of normal fasted rat parotid glands. The crystalloids varied in number from animal to animal and from lobule to lobule. They consisted of either parallel linear densities with a 5.5 nm periodicity, or of alternating major dense and minor light lines, with a 15 nm distance between successive major lines. It was concluded that the presence of a few such crystalloids in parotid glands of treated rats is not a cytopathic effect and they are significant only when present in large numbers.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/23717/1/0000689.pd

Fine structure of subcultivated stratified squamous epithelium

Subcultivated rat lingual epithelium derived from primary expiants remains mitotically active, possesses an organellar complement similar to the parent tissue, and undergoes terminal differentiation. Successful growth of primary cultures requires an incubation temperature below 34 [deg]C and the addition of dimethyl sulfoxide (DMSO) to the medium. The subcultures retain a stable morphological phenotype through a minimum of 15 passages. Cultures are long-lived and may be maintained for one year or more in any passage.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/23377/1/0000322.pd

Regional Nerve Block of the Temporomandibular Joint Capsule: A Technique for Clinical Research and Differential Diagnosis

In previous studies in which regional anesthesia of the temporomandibular joint capsule was used to examine the role of the joint in mandibular movement and distinguish it from muscle control, the anesthetic techniques used have not been satisfactorily described. The accuracy of the injeetion technique described in this paper was determined by dissection and radiographic examination of fixed and fresh specimens. Using this technique, trial patient studies were made using an anesthetic solution to which a radiopaque medium was added. Radiographic examination of the patients affirmed the location of the injected material, while clinical assessment determined its functional effectiveness. Using the described technique, anesthetic solution was accurately and reproducibly introduced posteriorly and laterally to the temporomandibular joint to achieve anesthesia of the joint.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/67376/2/10.1177_00220345800590110101.pd

Preliminary Assessment of the Efficacy of a T-Cell–Based Influenza Vaccine, MVA-NP+M1, in Humans

A single vaccination with MVA-NP+M1 boosts T-cell responses to conserved influenza antigens in humans. Protection against influenza disease and virus shedding was demonstrated in an influenza virus challenge study

Progression of Pathogenic Events in Cynomolgus Macaques Infected with Variola Virus

Smallpox, caused by variola virus (VARV), is a devastating human disease that affected millions worldwide until the virus was eradicated in the 1970 s. Subsequent cessation of vaccination has resulted in an immunologically naive human population that would be at risk should VARV be used as an agent of bioterrorism. The development of antivirals and improved vaccines to counter this threat would be facilitated by the development of animal models using authentic VARV. Towards this end, cynomolgus macaques were identified as adequate hosts for VARV, developing ordinary or hemorrhagic smallpox in a dose-dependent fashion. To further refine this model, we performed a serial sampling study on macaques exposed to doses of VARV strain Harper calibrated to induce ordinary or hemorrhagic disease. Several key differences were noted between these models. In the ordinary smallpox model, lymphoid and myeloid hyperplasias were consistently found whereas lymphocytolysis and hematopoietic necrosis developed in hemorrhagic smallpox. Viral antigen accumulation, as assessed immunohistochemically, was mild and transient in the ordinary smallpox model. In contrast, in the hemorrhagic model antigen distribution was widespread and included tissues and cells not involved in the ordinary model. Hemorrhagic smallpox developed only in the presence of secondary bacterial infections – an observation also commonly noted in historical reports of human smallpox. Together, our results support the macaque model as an excellent surrogate for human smallpox in terms of disease onset, acute disease course, and gross and histopathological lesions

Optimizing Staining Protocols for Laser Microdissection of Specific Cell Types from the Testis Including Carcinoma In Situ

Microarray and RT-PCR based methods are important tools for analysis of gene expression; however, in tissues containing many different cells types, such as the testis, characterization of gene expression in specific cell types can be severely hampered by noise from other cells. The laser microdissection technology allows for enrichment of specific cell types. However, when the cells are not morphologically distinguishable, it is necessary to use a specific staining method for the target cells. In this study we have tested different fixatives, storage conditions for frozen sections and staining protocols, and present two staining protocols for frozen sections, one for fast and specific staining of fetal germ cells, testicular carcinoma in situ cells, and other cells with embryonic stem cell-like properties that express the alkaline phosphatase, and one for specific staining of lipid droplet-containing cells, which is useful for isolation of the androgen-producing Leydig cells. Both protocols retain a morphology that is compatible with laser microdissection and yield RNA of a quality suitable for PCR and microarray analysis

Role of Inn1 and its interactions with Hof1 and Cyk3 in promoting cleavage furrow and septum formation in S. cerevisiae

Cytokinesis requires coordination of actomyosin ring (AMR) contraction with rearrangements of the plasma membrane and extracellular matrix. In Saccharomyces cerevisiae, new membrane, the chitin synthase Chs2 (which forms the primary septum [PS]), and the protein Inn1 are all delivered to the division site upon mitotic exit even when the AMR is absent. Inn1 is essential for PS formation but not for Chs2 localization. The Inn1 C-terminal region is necessary for localization, and distinct PXXP motifs in this region mediate functionally important interactions with SH3 domains in the cytokinesis proteins Hof1 (an F-BAR protein) and Cyk3 (whose overexpression can restore PS formation in inn1Δ cells). The Inn1 N terminus resembles C2 domains but does not appear to bind phospholipids; nonetheless, when overexpressed or fused to Hof1, it can provide Inn1 function even in the absence of the AMR. Thus, Inn1 and Cyk3 appear to cooperate in activating Chs2 for PS formation, which allows coordination of AMR contraction with ingression of the cleavage furrow