Using tDCS to facilitate motor learning in speech production: The role of timing.

There exists debate regarding the extent to which transcranial direct current stimulation (tDCS) can affect or enhance human behavior. Here, we examined a previously unexplored domain: speech motor learning. We investigated whether speech motor learning in unimpaired participants can be enhanced using a single-session tDCS experiment, and investigated whether the timing of tDCS relative to a behavioral task affected performance.

Altered lymphatic drainage after breast-conserving surgery and axillary node dissection: local recurrence with contralateral intramammary nodal metastases.

A review of the literature shows that lymphoscintigraphy and sentinel node biopsy are feasible in patients with previous breast and axillary surgery and could be especially warranted because in these patients, lymphatic drainage might not include the axillary basin. We report a case of a woman with recurrent breast cancer after breast-conserving surgery. The patient was found to have metastases in the contralateral intramammary lymph nodes.

Further evidence for AQP8 expression in the myoepithelium of rat submandibular and parotid glands.

Previously (Wellner et al., Pflugers Arch 441:49-56, 2000) we suggested that the localization of the aquaporins (AQPs) AQP5 and AQP8 in the apical and basolateral membranes of rat submandibular gland (SMG) acinar cells, respectively, provides for transcellular water flow during saliva formation. While the localization of AQP5 in this gland has been verified in several laboratories, there have been differing reports regarding AQP8 localization.

Modifying the NH2 and COOH termini of aquaporin-5: effects on localization in polarized epithelial cells.

To reengineer polarized epithelial cell functions directly in situ, or ex vivo in the fabrication of an artificial organ, it is necessary to understand mechanisms that account for polarized membrane sorting. We have used the aquaporins (AQPs), a family of homotetrameric water channel proteins, as model membrane proteins for this purpose. AQP monomers contain six transmembrane-spanning domains linked by five interconnecting loops, with the NH2 and COOH termini residing in the cytosol.

Localization of AQP5/AQP8 chimeras in MDCK-II cells: exchange of the N- and C-termini.

AQP5 and AQP8 possess targeting/retention motifs which mediate their localization to the apical and basolateral membranes, respectively, of polarized MDCK-II cells. As targeting/retention motifs have been localized to the N- or C-termini of other AQPs, we sought the location of such motifs in AQPs 5 and 8 by exchanging their corresponding N- or C-termini and examining the expression, localization, and function of the resultant chimeras. We did not detect the expression of constructs in which the C-terminus of AQP5 was replaced by the C-terminus of AQP8.

Primary culture of polarized human salivary epithelial cells for use in developing an artificial salivary gland.

Therapeutic irradiation for head and neck cancer, and the autoimmune disease Sjogren's syndrome, lead to loss of salivary parenchyma. They are the two main causes of irreversible salivary gland hypofunction. Such patients cannot produce adequate levels of saliva, leading to considerable morbidity.

Increased fluid secretion after adenoviral-mediated transfer of the human aquaporin-1 cDNA to irradiated miniature pig parotid glands.

The treatment of most head and neck cancer patients includes ionizing radiation (IR). Salivary glands in the IR field suffer irreversible damage. Previously, we reported that adenoviral (Ad)-mediated transfer of the human aquaporin-1 (hAQP1) cDNA to rat submandibular glands following IR restored salivary flow to near normal levels.

The stable nitroxide tempol facilitates salivary gland protection during head and neck irradiation in a mouse model.

Purpose: Radiotherapy is commonly used to treat a majority of patients with head and neck cancers. The long-term radiation-induced reduction of saliva output significantly contributes to the posttreatment morbidity experienced by these patients. The purpose of this study was to test the ability of the stable-free radical Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl), an established radioprotector, to prevent radiation-induced salivary hypofunction in mice.

The effect of body mass index on complications from cardiac surgery in the oldest old.

Objectives: Overweight (body mass index (BMI)>25) and obesity (BMI>30) are associated with adverse outcomes in the general population. In older people, an increased risk of adverse events was observed in leaner and overweight older subjects. We evaluated the association between BMI and cardiac surgery complications in subjects aged 75 and older.

Long-term survival and quality of life in cardiac surgical patients with prolonged intensive care unit length of stay.

Background: Patients with prolonged intensive care unit (ICU) stays after cardiac operations are labor intensive and expensive. We sought to determine whether exhaustive ICU efforts result in survival or quality-of-life benefits and whether outcome could be predicted.

Methods: We retrospectively analyzed all adult cardiac surgical patients in 1998 for ICU stays more than 14 days.

Gene transfer to salivary glands.

This article provides a review of the application of gene transfer technology to studies of salivary glands. Salivary glands provide an uncommon target site for gene transfer but offer many experimental situations likely of interest to the cell biologist. The reader is provided with a concise overview of salivary biology, along with a general discussion of the strategies available for gene transfer to any tissue.

Recombinant adeno-associated virus serotype 2 vectors mediate stable interleukin 10 secretion from salivary glands into the bloodstream.

We have constructed a recombinant adeno-associated virus serotype 2 vector encoding human interleukin 10 (rAAVhIL10). IL-10 is a potent antiinflammatory/immune cytokine, which has received growing attention for its therapeutic potential. Human IL-10 (hIL-10) production was virus dose dependent after in vitro infection of HSG cells, a human submandibular gland cell line.

Polarized sorting of aquaporins 5 and 8 in stable MDCK-II transfectants.

Localization of aquaporin (AQP) water channels to either apical or basolateral membranes is important for various epithelial functions. We have established MDCK-II cell transfectants stably expressing AQP5 (RW5 cells) or AQP8 (RW8 cells). The expression of both AQPs was confirmed by the results of immunofluorescence microscopy and immunoblot analysis.

Cationic liposome-mediated gene transfer to rat salivary epithelial cells in vitro and in vivo.

Background: Previously we have shown that gene transfer to salivary gland epithelial cells readily occurs via recombinant adenoviruses, although the response is short-lived and results in a potent host immune response. The aim of the present study was to assess the feasibility of using cationic liposomes to mediate gene transfer to rat salivary cells in vitro and in vivo.

Methods: Initially, for transfection in vitro, we used two cationic liposome formulations (GAP-DLRIE/DOPE and DOSPA/DOPE) complexed with plasmid encoding human growth hormone (hGH) as a reporter gene.

Evidence that aquaporin-8 is located in the basolateral membrane of rat submandibular gland acinar cells.

It is possible that, during primary saliva formation, aquaporins (AQPs) facilitate transcellular water flow across acinar cells to the lumina of salivary glands. In the rat submandibular gland (rSMG) AQP5 is localized in the apical membranes of acinar cells. The presence of a basolateral AQP in the same cell type has not been reported.

Salivary glands as a model for craniofacial applications of gene transfer.

The potential applications of gene transfer technology to all branches of medicine are increasing. It is quite likely that within the next 10-20 years surgical practice routinely will utilize gene transfer, at least adjunctively. The purpose of this review is to familiarize the oral and maxillofacial surgeon with this technology.

Construction and function of a recombinant adenovirus encoding a human aquaporin 1-green fluorescent protein fusion product.

Transfer of the human aquaporin 1 (hAQP1) gene provides a novel way to potentially correct the severe salivary hypofunction associated with therapeutic radiation for head and neck cancer. To facilitate the study of individual cells transduced with this gene, we have designed a fusion product of the hAQP1 and jellyfish green fluorescent protein (GFP) cDNAs. An expression plasmid, pACCMVhAQP1GFP, and a recombinant adenovirus, AdhAQP1GFP, encoding this fusion product were constructed.

Trp1, a candidate protein for the store-operated Ca(2+) influx mechanism in salivary gland cells.

The trp gene family has been proposed to encode the store-operated Ca(2+) influx (SOC) channel(s). This study examines the role of Trp1 in the SOC mechanism of salivary gland cells. htrp1, htrp3, and Trp1 were detected in the human submandibular gland cell line (HSG).

Polarized secretion of transgene products from salivary glands in vivo.

Previously (Kagami et al. Hum. Gene Ther.

Construction and in vivo efficacy of a replication-deficient recombinant adenovirus encoding murine growth hormone.

We have constructed a recombinant, replication-deficient, first-generation adenovirus-encoding mouse GH (mGH), AdCMVmGH. This virus directed mGH production from an epithelial cell line in vitro in a dose-dependent manner. When injected into the quadriceps muscle or submandibular ducts of mGH-deficient Snell dwarf mice, AdCMVmGH resulted in the production of significantly elevated serum mGH levels.

Relationship between adenovirus-mediated aquaporin 1 expression and fluid movement across epithelial cells.

AdhAQP1, a recombinant adenovirus encoding the human water channel aquaporin 1 (AQP1), has been shown to be useful for gene therapy of salivary glands rendered hypofunctional following irradiation. Here we utilized AdhAQP1 to examine the relationship between AQP1 expression and fluid movement across a polarized salivary epithelial cell monolayer. In response to a 440 to 340 mosm gradient, net fluid movement across cells infected with AdhAQP1 was approximately 10-fold that seen in uninfected cells or cells infected with a control virus.

Systemic action of human growth hormone following adenovirus-mediated gene transfer to rat submandibular glands.

We have previously suggested that although salivary glands function in an exocrine manner they might none the less offer a useful way to deliver therapeutic proteins systemically. As a direct functional test of this hypothesis, we constructed a recombinant adenovirus (AdCMVhGH) encoding human growth hormone (hGH) and then studied the biological action of hGH produced following transfer of the hGH gene to rat submandibular glands. At 48 h following infusion of AdCMVhGH into these glands via cannulation of the main excretory duct, serum levels of hGH were approximately 16 ng/ml and rat insulin-like growth factor-1 was elevated approximately 25%.

In vitro fusion of endocytic vesicles: effects of reagents that alter endosomal pH.

Ricin, a plant toxin that binds to galactose-terminated glycoproteins and glycolipids on the cell surface, is internalized into endosomes before reaching the cytosol where it exerts its toxic activity. Fusion of early endosomes containing ricin or transferrin was demonstrated by using postnuclear supernatant fractions from K-562 cells. For both ligands, fusion depended on time, temperature, and ATP and was blocked by preincubation with N-ethylmaleimide.

Anti-salivary antibodies in primary Sjögren's syndrome.

Earlier studies have described an antibody that recognized salivary ductal epithelium in sera from 15-50% of patients with primary Sjögren's syndrome; however, the specific salivary antigen in those studies was not identified. The present study further investigated this unknown salivary antigen. Twenty-nine of 31 patients (94%) with primary Sjögren's syndrome demonstrated IgG antinuclear antibodies that bound to an epithelial cell line with ductal characteristics derived from a human salivary gland.