Stimulated saliva composition in patients with cancer of the head and neck region

Background To analyse over time changes in stimulated whole saliva regarding total protein, Immunoglobulin A (IgA), and mucin type O-glycans (mostly MUC5B and MUC7) in head and neck cancer patients. Methods 29 dentate patients (20 men and 9 women, 59 ± 8 years) treated with curative radiation therapy and chemotherapy for cancer of the head and neck region were included. The stimulated whole salivary secretion rate was determined and saliva collected at four time-points: at pretreatment, and at 6 months, 1 and 2 years post treatment. The total protein concentration was determined spectrophotometrically by using Bicinchoninic Acid assay and Immunoglobulin A (IgA) by using ELISA technique. Glycosylation pattern of salivary mucins was determined in samples collected pre- and post treatment by using LC/MS electrospray and mucin content quantified using SDS-AgPAGE gels and PAS staining. Results Compared with pretreatment, the total protein concentration was increased already at 6 months post treatment (p < 0.01), and continued to increase up to 2 years post treatment (p < 0.001). During that period no significant changes in IgA concentration was detected. At pretreatment, the output/min of both total protein and IgA was significantly higher than at all time-points post treatment. Saliva from the cancer patients showed a low abundance/no detectable MUC7, while the MUC5B level remained, compared to saliva from a healthy control. The glycomic analysis showed that the percentage of core 2 O-glycans was increased as core 1, 3 and 4 O-glycans were decreased. The level of sialylation was higher at 6 months post treatment, while sulfation was lower. Conclusion A decreased output per minute of proteins at decreased salivary secretion rate, as well as reduced sulfation of MUC5B at 6 months post treatment tended to correlate with the patients’ experience of sticky saliva and oral dryness. At 2 years post treatment, the decreased amount of IgA combined with a lowered salivary secretion rate indicate a reduced oral defense with increased risk of oral infections.

CSF Secretion Is Not Altered by NKCC1 Nor TRPV4 Antagonism in Healthy Rats

Background: Cerebrospinal fluid (CSF) secretion can be targeted to reduce elevated intracranial pressure (ICP). Sodium-potassium-chloride cotransporter 1 (NKCC1) antagonism is used clinically. However, supporting evidence is limited. The transient receptor potential vanilloid-4 (TRPV4) channel may also regulate CSF secretion and ICP elevation. We investigated whether antagonism of these proteins reduces CSF secretion. Methods: We quantified CSF secretion rates in male Wistar rats. The cerebral aqueduct was blocked with viscous mineral oil, and a lateral ventricle was cannulated. Secretion rate was measured at baseline and after antagonist administration. Acetazolamide was administered as a positive control to confirm changes in CSF secretion rates. Results: Neither NKCC1, nor TRPV4 antagonism altered CSF secretion rate from baseline, n = 3, t(2) = 1.14, p = 0.37, and n = 4, t(3) = 0.58, p = 0.6, respectively. Acetazolamide reduced CSF secretion by ~50% across all groups, n = 7, t(6) = 4.294, p = 0.005. Conclusions: Acute antagonism of NKCC1 and TRPV4 proteins at the choroid plexus does not reduce CSF secretion in healthy rats. Further investigation of protein changes and antagonism should be explored in neurological disease where increased CSF secretion and ICP are observed before discounting the therapeutic potential of protein antagonism at these sites.

Effects of Energy Deficit on Secretory IgA During a Simulated Multi-Stressor Military Operation

Objectives Secretory IgA (SIgA) is a critical component of mucosal immunity and a first line of defense against pathogens. Intense physical exercise, lack of sleep, and inadequate energy intake are frequently observed during military training and operations. These factors are associated with a decline in SIgA and may increase the risk of infection; however, to what degree each of these factors contributes to immune dysfunction is unclear. This study aimed to determine the effect of severe energy deficit on mucosal immunity (SIgA) during a multi-day period of intense training. Methods The parent study was a randomized, crossover trial in healthy males (n = 10, 22.4 ± 5.4 y, 87.3 ± 10.9 kg) to assess the effect of severe negative energy balance on inflammation, iron absorption, and other physiological and cognitive outcomes during a simulated sustained military operation (SUSOPS; high energy expenditure with repeated bouts of intense exercise). Participants completed two SUSOPS trials and were randomized to consume ± 10% of estimated total daily energy expenditure (TDEE, energy balance) or 45% of TDEE (energy deficit). At 0500 on each SUSOPS day (D1: baseline, D2:24 h, D3:48 h), participants placed polyester oral swabs under their tongue for 3-mins. A second swab was collected (i.e., placed under the tongue until saturation) to ensure adequate sample volume. SIgA secretion rate (μg/min) was calculated from SIgA concentration (μg/mL; enzyme-linked immunosorbent assay) and salivary flow rate (mL/min). Dependent variables were log10 transformed due to non-normal distribution and data were analyzed using linear mixed models. Results Independent of treatment, a main effect of time (P = 0.01) was observed where SIgA secretion rate declined by 20% from D2 [1.77 ± 0.34 μg/min] to D3 [1.41 ± 0.51 μg/min], P = 0.001, with no significant treatment by time interactions. A main effect of time (P = 0.01) was also found wherein SIgA concentration declined by 13% from D2 [2.67 ± 0.32 μg/mL] to D3 [2.33 ± 0.37 μg/mL], P = 0.001. There were no main or treatment effects with regard to SIgA flow rate. Conclusions Mucosal immune response, as measured by SIgA, declined in response to SUSOPS. Severe energy deficit did not exacerbate the decline in SIgA secretion rate observed in response to the high intensity, multi-stressor training scenario. Funding Sources US Army Medical Research and Development Command.

Effects of indacaterol on the LPS-evoked changes in fluid secretion rate and pH in swine tracheal membrane

An acquired dysregulation of airway secretion is likely involved in the pathophysiology of chronic bronchitis and chronic obstructive pulmonary disease (COPD). Nowadays, it is widely known that several kinds of long-acting bronchodilators reduce the frequency of COPD exacerbations. However, limited data are available concerning the complementary additive effects on airflow obstruction. Using an optical method and a selective pH indicator, we succeeded in evaluating the gland secretion rate and the pH in swine tracheal membrane. A physiologically relevant concentration of acetylcholine (ACh) 100 nM induced a gradual increase in the amount of gland secretion. Lipopolysaccharides (LPS) accelerated the ACh-induced secretory responses up to around threefold and lowered the pH level significantly. Long-acting β2-agonists (LABAs) including indacaterol (IND), formoterol, and salmeterol restored the LPS-induced changes in both the hypersecretion and acidification. The subsequent addition of the long-acting muscarine antagonist, glycopyrronium, further increased the pH values. Two different inhibitors for cystic fibrosis transmembrane conductance regulator (CFTR), NPPB and CFTRinh172, abolished the IND-mediated pH normalization in the presence of both ACh and ACh + LPS. Both immunofluorescence staining and western blotting analysis revealed that LPS downregulated the abundant expression of CFTR protein. However, IND did not restore the LPS-induced decrease in CFTR expression on Calu-3 cells. These findings suggest that the activation of cAMP-dependent HCO3− secretion through CFTR would be partly involved in the IND-mediated pH normalization in gland secretion and may be suitable for the maintenance of airway defense against exacerbating factors including LPS.

The ratio of roughage to concentrate on milk secretion rate in goats fed Stylosanthes ha mata hay

An experiment was conducted to evaluate the effect of roughage to concentrate ratio (R:C) on milk secretion rate (g/h) in goats using a cross-over design in which each goat passed sequentially through all the treatments in random order. The treatments (R: C) were: A = 70 percent roughage : 30 percent concentrate; B = 50 percent roughage : 50 percent concentrate e and C = 30 percent roughage : 70 percent concentrate. In the experiment, seven goats were milked once a day, from the 2nd to 8th week of lactation, thrice per week for 2 weeks per treatment. The animals were fed at 4 percent body weight (DM basis). The feed consisted of Stylosanthes hamata hay (S. hamata hay) and a 17.2 percent CP concentrate ration. Data were analysed using the treatments as main effect with live weight and week of lactation as covariates. There was no significant treatment effects on mülk secretion rate (P>0.05). Overall milk secretion rate was 5.23gh. Week of lactation also had no significant effect (P>0.05). However, live weight of doe at time of milking had a highly significant effect (P>0.001) with milk secretion rate increasing by 0.41g/h per kilogram live weight. The R2 values for the predictive mathematical relationship for goats fed Stylosanthes hamata hay at 4 percent body weight in the dry season was R2 = 0.888. It can therefore be deduced that for a ladating doe in the 5th week of lactation at 20kg body weight fed at 4 percent with Stylosanthes hamata hay to concentrate ratio at 70 percent roughage : 30 percent concentrate would give a milk secretion rate of 6.717g/h while at 50 percent roughage : 50 percent concentrate it would give 6.662 g/h and 30 percent roughage : 70 percent concentrate level would give 7.054 g/h. Keywords:,,,