Identification of Salivary Metabolic Signatures Associated with Primary Sjögren's Disease.

Sjögren's disease (SjD) is the second most prevalent autoimmune disorder that involves chronic inflammation of exocrine glands. Correct diagnosis of primary SjD (pSjD) can span over many years since disease symptoms manifest only in advanced stages of salivary and lachrymal glandular destruction, and consensus diagnostic methods have critical sensitivity and selectivity limitations. Using nuclear magnetic resonance (NMR) spectroscopy, we determined the composition of metabolites in unstimulated saliva samples from 30 pSjD subjects and 30 participants who do not have Sjögren's disease (non-Sjögren's control group, NS-C). Thirty-four metabolites were quantified in each sample, and analysis was conducted on both non-normalized (concentration) and normalized metabolomics data from all study participants (ages 23-78) and on an age-restricted subset of the data (ages 30-70) while applying false discovery rate correction in determining data significance. The normalized data of saliva samples from all study participants, and of the age-restricted subset, indicated significant increases in the levels of glucose, glycerol, taurine, and lactate, as well as significant decreases in the levels of 5-aminopentanoate, acetate, butyrate and propionate, in subjects with pSjD compared to subjects in the NS-C group. Additionally, a significant increase in choline was found only in the age-restricted subset, and a significant decrease in fucose was found only in the whole study population in normalized data of saliva samples from the pSjD group compared to the NS-C group. Metabolite concentration data of saliva samples from all study participants, but not from the age-restricted subset, indicated significant increases in the levels of glucose, glycerol, taurine, and lactate in subjects with pSjD compared to controls. The study showed that NMR metabolomics can be implemented in defining salivary metabolic signatures that are associated with disease status, and can contribute to differential analysis between subjects with pSjD and those who are not affected with this disease, in the clinic.

Delineating cell behavior and metabolism of non-melanoma skin cancer in vitro.

Non-melanoma skin cancers - basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) - are the most frequent forms of malignant neoplasm in humans worldwide. The etiology of these carcinomas is multifactorial. In addition to the harmful effect of UV light, altered cross-talk between neoplastic epithelial cells and the supporting dermal fibroblasts contributes to the regulation of tumor cell behavior, growth and survival. Metabolic cooperation between these cell types allows them to adapt and react to changes in their surrounding microenvironment by modifying their cellular bioenergetics and biosynthesis. We characterized the growth, behavior, and metabolic activity of human BCC cells, E-cadherin-competent SCC cells and E-cadherin-suppressed SCC cells in the presence or absence of dermal fibroblasts. In mono-cultures and co-cultures, BCC and SCC cells demonstrated distinct morphology, growth and organizational patterns. These tumor cells also exhibited unique patterns of consumption and secretion profiles of glucose, lactate, acetate, glutamine, glutamate, and pyruvate. In comparison to mono-cultures, growth of fibroblasts with either BCC cells or SCC cells enriched the cell growth environment, allowed for metabolic cooperation between these two cell types, and resulted in alterations in the metabolic profiles of the co-cultures. These alterations were affected by the cancer cell type, culture confluence and the composition of the growth medium. Characterizing the bioenergetics of BCC and SCC cells in the context of tumor-stromal interactions is not only important for further understanding of tumor pathogenesis, but also can illuminate potential new targets for novel, metabolic-based therapies for non-melanoma skin cancers.

Angiotensin II evokes calcium-mediated signaling events in isolated dog pancreatic epithelial cells.

INTRODUCTION: Calcium-activated chloride conductance has been identified in normal pancreatic duct cells. Recent in vitro evidence suggests that angiotensin II (AngII) stimulates pancreatic secretion in both cystic fibrosis (CFPAC) and transformed pancreatic cells. AIMS: To investigate calcium-mediated stimulatory effects of AngII in both nontransformed dog pancreatic duct epithelial (DPDE) and CFPAC cells. METHODS: Western blots were performed in both cells seeking AngII receptors. In additional studies, DPDE and CFPAC cells were grown on vitrogen-coated glass cover slips and loaded with Indo-1-AM dye. Cells were placed in a confocal microscope's perfusion chamber and perfused with 100 microM AngII or ATP (control). Cells were excited with UV light, and intracellular calcium ([Ca+2]i) was read using fluorescence emission at 405 and 530 nm. Finally, single channels in the DPDE cells were examined using cell-attached patch clamps. Current amplitude histograms provided estimates of the conductance and open probability of channels. RESULTS: Western blots demonstrated presence of both AT and AT AngII receptors in DPDE and CFPAC cells; the density of AT receptors appeared lower than that of AT receptors. Basal intracellular calcium concentrations did not differ between DPDE (109 +/- 11 nM) and CFPAC (103 +/- 8 nM) cells. AngII significantly increased measured intracellular calcium concentrations in both DPDE (909 +/- 98 nM) and CFPAC (879 +/- 207 nM) cells, as did ATP (DPDE = 1722 +/- 228 nM; CFPAC = 1522 +/- 245 nM). In the patch clamp studies, a variety of different channels were observed; they appeared to be an 11pS nonselective cation (NSC) channel, a 4.6pS Na+ channel, a 3pS anion channel, and an 8pS chloride channel. The latter channel had characteristics similar to cystic fibrosis transmembrane conductance regulator (CFTR). Apical or basolateral application of AngII activated both the 11pS NSC and the 3pS channels. CONCLUSION: In nontransformed DPDE and CFPAC cells, specific AngII receptors mediate increases in [Ca ]. The latter effect of AngII may elicit activation of calcium-mediated chloride channels, suggesting a role for AngII as an alternative mediator of pancreatic ductal secretion.

Dose and wavelength of laser light have influence on the repair of cutaneous wounds.

OBJECTIVE: The objective of the present study was to compare histologically the effect of GaAlAs (lambda 830 nm, phi approximately 2 mm(2), 35 mW) and InGaAlP (lambda 685 nm, phi approximately 2 mm(2), 35 mW) lasers, alone or in association with doses of 20 or 50 J/cm(2) on cutaneous wounds in the dorsum of the Wistar rat. BACKGROUND DATA: The healing time of surgical wounds is of extreme importance and it is usually associated with a post-operative period free of infection and with less pain and inflammation. MATERIALS AND METHODS: Sixty Wistar rats were divided into seven groups: Group I - control (non-irradiated); Group II - lambda 685 nm, 20 J/cm(2); Group III - lambda 830 nm, 20 J/cm(2); Group IV - lambda 685 nm and lambda 830 nm, 20 J/cm(2); Group V - lambda 685 nm, 50 J/cm(2)); Group VI - lambda 830 nm, 50 J/cm(2); and Group VII - lambda 685 nm and 830 nm, 50 J/cm(2). The animals were sacrificed 3, 5, and 7 days after surgery. RESULTS: Light microscopic analysis using H&E and Picrosírius stains showed that, at the end of the experimental period, irradiated subjects showed increased collagen production and organization when compared to non-irradiated controls. Inflammation was still present in all groups at this time. CONCLUSION: Group IV (lambda 830 nm and lambda 685 nm, 20 J/cm(2)) presented better results at the end of the experimental period. It is concluded that low-level light therapy (LLLT) can have a positive biomodulatory effect on the repair of cutaneous wounds.