LPLI attenuates apoptosis.

<p>Bax and cleaved caspase-3 were assessed by immunohistochemistry. There is no expression of Bax or cleaved caspase-3 in control SMGs. Both markers were localized in SMGs from the diabetic rats, and were strongly stained in striated duct cell cytoplasm (arrows), but after LPLI there was no expression of Bax and only a faint expression of cleaved caspase-3. C, Control; D, Diabetes; DL, Diabetes + LPLI; ED, excretory duct; SD, striated duct. Scale bars, 50 μM.</p

LPLI decreases the expression of cell death markers.

<p>Western blotting analysis showed that inflammatory markers (HMGB1, AGE, RAGE and phospho NF-κB p65) and apoptotic markers (Bax, p53, phospho-p53 (Ser15) and cleaved caspase-3) were increased in diabetic rats and were reduced by LPLI. Protein sizes were evaluated by standard protein markers, and their sizes were as follows: HMGB1 (25 kDa), AGE (non-specified), RAGE (46 kDa), phospho NF-κB p65 (65kDa), Bax (21 kDa), phospho-p53 (Ser15) (53 kDa), p53 (53 kDa), cleaved caspase-3 (17–19 kDa) and β-actin (42 kDa). TNF-α and Bad were assessed by immunohistochemistry. There was no expression of TNF-α or Bad in the control group. TNF-α and Bad were both strongly stained in striated duct cell cytoplasm (arrows) in SMGs from diabetic rats, but can barely be seen after LPLI (Fig 4). C, Control; D, Diabetes; DL, Diabetes + LPLI; ED, excretory duct; SD, striated duct. Scale bars, 50 μM.</p

Metabolism patterns.

<p>Metabolism patterns.</p

Experimental design.

<p>The rats were divided into 3 groups: control (C), diabetes (D) and diabetes + LPLI (DL). On day 1, diabetes was induced in the D and DL groups by injection of STZ. On day 29, the DL group received the LPLI. On day 30, all rats were sacrificed and their SMGs were collected. Each rat’s blood glycemia level was checked on days 3 and 30.</p

Co-localization of Bax and HMGB1 assessed by double immunofluorescence staining.

<p>Double-positive cells were faintly localized in striated and excretory duct cells in the control SMGs, with intense staining in duct cells, as well as in endothelial cells of diabetic rat SMGs. These expression patterns are attenuated by LPLI. C, Control; D, Diabetes; DL, Diabetes + LPLI; ED, excretory duct; SD, striated duct; EN, endothelium. Original magnification 20X.</p

LPLI attenuates inflammation via NF-κB.

<p>The expression of AGE, RAGE and phospho NF-κB p65 is observed only in diabetic rats (arrows), in striated and excretory duct cell cytoplasm, with little expression of AGE and RAGE in the DL group and no expression of phospho NF-κB after LPLI. C, Control; D, Diabetes; DL, Diabetes + LPLI; ED, excretory duct; SD, striated duct; ID, intercalated duct. Scale bars, 50 μM.</p

LPLI reduces HMGB1 accumulation in SMGs from diabetic rats.

<p>Morphology of SMGs from control, diabetic and diabetic rats treated with LPLI shows typical acinar and ducts cells in all 3 groups. Immunohistochemical analysis of HMGB1 in control, diabetic and diabetic rats treated with LPLI. HMGB1 expression was localized in the nuclei of striated duct cells (arrows), as well as, in the nuclei of endothelial cells in SMGs of diabetic rats. C, Control; D, Diabetes; DL, Diabetes + LPLI; ED, excretory duct; SD, striated duct. Scale bars, 50 μM. Diabetes increases RAGE and HMGB1 mRNA levels analyzed by qRT-PCR, and LPLI reduces those levels to patterns similar to the control group. Two independent measurements were averaged and relative gene expression levels were calculated as a ratio to β-actin expression of each sample.</p

Osteoradionecrosis of the jaws: case series treated with adjuvant low-level laser therapy and antimicrobial photodynamic therapy

<div><p>Abstract Background: Osteoradionecrosis of the jaw (ORNJ) is the most severe and complex sequel of head and neck radiotherapy (RT) because of the bone involved, it may cause pain, paresthesia, foul odor, fistulae with suppuration, need for extra oral communication and pathological fracture. We treated twenty lesions of ORNJ using low-level laser therapy (LLLT) and antimicrobial photodynamic therapy (aPDT). The objective of this study was to stimulate the affected area to homeostasis and to promote the healing of the oral mucosa. Methods: We performed aPDT on the exposed bone, while LLLT was performed around the bone exposure (red spectrum) and on the affected jaw (infrared spectrum). Monitoring and clinical intervention occurred weekly or biweekly for 2 years. Results: 100% of the sample presented clinical improvement, and 80% presented complete covering of the bone exposure by intact oral mucosa. Conclusion: LLLT and aPDT showed positive results as an adjuvant therapy to treat ORNJ.</p></div

The effect of heparin on steady-state FRET-peptide hydrolysis by HNE.

<p>The influence of heparin concentration upon steady-state HNE kinetics parameters <i>k</i>_cat [A] and <i>K</i>_M [B] was determined spectrofluometricallyas described under “Experimental Procedures”. The fluorescence increase of FRET-peptide hydrolysis (0.2 - 10 µM) by 12.6 nM HNE were performed in 10 mM Hepes, pH 7.4, containing 140 mM NaCl and 0,05% Triton X-100 at 37°C.</p

Stopped-flow time-traces of FRET-peptidehydrolysis by HNE in the presence of heparin.

<p><i>A</i>, the stopped-Flow fluorescence kinetic recording of 3.8 µMFRET-peptide hydrolysis by 12.6 nM HNEperformed at 25°C in 10 mM Tris-HCl buffer, pH 7.4, containing 100 mM NaCl supplemented with 50 µM heparin. The progress of the reaction was monitored by the fluorescence increase of the released product recorded on two adjacent time regions with distinct sampling periods: 0.5 ms from 0 to 2 s, 2 ms from 2 to 4 s. Gray solid line represents the best fit deduced from the mechanism depicted in Scheme I in the presence of heparin using DynaFit IV® Software [see Experimental Procedures]. The insert graphic represents the associate residual errors from the best fit curve with experimental data. <i>B</i>, the HNE species in function of time reaction in presence of heparin: complex enzyme-heparin, EH (<b>–</b>); complex enzyme-substrate-heparin, ESH <b>(–</b> •<b>–</b>) and complex acyl-enzyme-heparin, ES'H (<b>- - -</b>).</p