Strategies of use of a specific immunoglobulin-rich egg yolk powder in weaning piglets

Two experiments were performed to determine the best strategy of use of the product TRACTcare® 4P (ITPSA) (TC, specific immunoglobulin-rich egg yolk powder within an energetic fatty acid matrix) in piglets from weaning and for 6 weeks, in diets without or with inclusion of antibiotics. Each trial was performed with 144 piglets in 24 pens, in a completely randomized design blocked by initial body weight. Feeds were formulated according to animal requirements in two periods. In the first trial no antibiotics were included in the feeds and no room disinfection from previous trial was performed; treatments were: 1) Negative control (NC); 2) NC+TC on top of the feed within the hopper for the first 3 days on trial (30 g/pig×day), and eventually if diarrhea appeared (TCOT); 3) NC+TC ad libitum provided in an extra hopper within the pen (TCAL); and 4) NC+TC at 5 g/kg added to the feed in the mixer (TC5). In the second trial, treatments were: 1) Positive control: basal diet that included 250 mg/kg amoxiciline (BD)+100 mg/kg colistine (AC); 2) BD+2 g/kg TC (TC2A); 3) BD+5 g/kg TC (TC5A); and 4) BD+8 g/kg TC (TC8A). In diets without antibiotics, the product TC at 5 g/kg in the feed numerically improved BW by 8% compared to Control animals, while G:F was almost identical between both groups. When antibiotics were used in the feed, replacement of colistin at 100 mg/kg for TC at 2 g/kg in feed numerically improved the performance compared to Positive control animals (for the whole trial period ADG 8% better: 390 g vs. 361 g; G:F 1% better: 0.748 kg/kg vs. 0.742 kg/kg), possibly due to the stimulation of feed consumption at weaning. In both trials, the lower number of dead and culled animals from TC5 and TC2A together with higher BW represented an advantage over Control treatments of 6% to 10% animals more and 15% to 17% total BW more at the end of the trial.

Gene transfer into the airway epithelium of animals by targeting the polymeric immunoglobulin receptor.

Genes of interest can be targeted specifically to respiratory epithelial cells in intact animals with high efficiency by exploiting the receptor-mediated endocytosis of the polymeric immunoglobulin receptor. A DNA carrier, consisting of the Fab portion of polyclonal antibodies raised against rat secretory component covalently linked to poly-L-lysine, was used to introduce plasmids containing different reporter genes into airway epithelial cells in vivo. We observed significant levels of luciferase enzyme activity in protein extracts from the liver and lung, achieving maximum values of 13,795 +/- 4,431 and 346,954 +/- 199,120 integrated light units (ILU) per milligram of protein extract, respectively. No luciferase activity was detected in spleen or heart, which do not express the receptor. Transfections using complexes consisting of an irrelevant plasmid (pCMV lacZ) bound to the bona fide carrier or the expression plasmid (pGEMluc) bound to a carrier based on an irrelevant Fab fragment resulted in background levels of luciferase activity in all tissues examined. Thus, only tissues that contain cells bearing the polymeric immunoglobulin receptor are transfected, and transfection cannot be attributed to the nonspecific uptake of an irrelevant carrier-DNA complex. Specific mRNA from the luciferase gene was also detected in the lungs of transfected animals. To determine which cells in the lungs are transfected by this method, DNA complexes were prepared containing expression plasmids with genes encoding the bacterial beta-galactosidase or the human interleukin 2 receptor. Expression of these genes was localized to the surface epithelium of the airways and the submucosal glands, and not the bronchioles and alveoli. Receptor-mediated endocytosis can be used to introduce functional genes into the respiratory epithelium of rats, and may be a useful technique for gene therapy targeting the lung.

Dissociation from BiP and Retrotranslocation of Unassembled Immunoglobulin Light Chains Are Tightly Coupled to Proteasome Activity

Unassembled immunoglobulin light chains expressed by the mouse plasmacytoma cell line NS1 (KNS1) are degraded in vivo with a half-life of 50-60 min in a way that closely resembles endoplasmic reticulum (ER)-associated degradation (Knittler et al., 1995). Here we show that the peptide aldehydes MG132 and PS1 and the specific proteasome inhibitor lactacystin effectively increased the half-life of KNS1, arguing for a proteasome-mediated degradation pathway. Subcellular fractionation and protease protection assays have indicated an ER localization of KNS1 upon proteasome inhibition. This was independently confirmed by the analysis of the folding state of KNS1and size fractionation experiments showing that the immunoglobulin light chain remained bound to the ER chaperone BiP when the activity of the proteasome was blocked. Moreover, kinetic studies performed in lactacystin-treated cells revealed a time-dependent increase in the physical stability of the BiP-KNS1complex, suggesting that additional proteins are present in the older complex. Together, our data support a model for ER-associated degradation in which both the release of a soluble nonglycosylated protein from BiP and its retrotranslocation out of the ER are tightly coupled with proteasome activity.

Papillary cystoadenoma lymphomatosum (Warthin-like) of minor salivary glands

Papillary cystadenoma lymphomatosum is a benign salivary gland tumor most frequently located in the parotid gland (Warthin"s tumor). Its presentation in other major, or in minor, salivary glands is rare. Clinically, it manifests as a slow growing tumor, fluctuant on palpation due to its cystic morphology. The treatment of choice is complete excision with wide tumor-free margins. We present a 73-year-old female patient with an asymptomatic tumor of 8 years evolution in the right posterior area of the hard palate. We performed surgical excision and a biopsy, which was reported as papillary cystadenoma lymphomatosum. During the post-operative examination carried out after 3 weeks, it was observed that the lesion had recurred. The lesion was re-operated, performing the excision with CO2 laser and including the periosteum to ensure complete resection of the tumor. At 10 months follow-up, there was no recurrence of the lesion. This article includes a review of this condition and discusses its most important clinical and pathologic features and therapeutic approaches.

Minor salivary gland tumors: a clinicopathological study of 18 cases

Introduction: Minor salivary gland tumors (MSGTs) are infrequent, representing 10-15% of all salivary neoplasms. Despite this low frequency, MSGTs conform a heterogeneous group of neoplasms characterized by a broad range of histological types. Patients and method: We identified cases of MSGT in a retrospective study of the biopsies made in the period 1997-2007 in the Service of Oral Surgery (Dental Clinic of the University of Barcelona, Spain). The data collected comprised patient age and sex, the clinical characteristics and location of the tumor, the duration of the lesion, its size, the treatment provided, and the histopathological findings. Results: Of the 18 cases of MSGT studied, 12 corresponded to women (66.7%) and 6 to men (33.3%). The great majority (94.4%) were benign tumors. The preferential location was the posterior third of the hard palate (33.2%), followed by the soft palate (16.7%) and the mucosa of the upper lip (16.7%). The histopathological diagnoses of our MSGTs comprised 10 pleomorphic adenomas (55.3%), 2 cystadenomas (11.1%), 1 myoepithelioma (5.6%), 1 sialadenoma papilliferum (5.6%), 1 basal cell adenoma (5.6%), 1 Warthin"s tumor (5.6%), 1 canalicular adenoma (5.6%), and 1 low-grade polymorphic adenocarcinoma (5.6%). Discussion and conclusions: Coinciding with our own results, the literature describes a high recurrence rate for MSGTs (5-30%) when surgical removal is incomplete. Six percent of all benign minor salivary gland tumors are considered to relapse, versus 65% of all malignant lesions. Periodic clinical controls are required, since the possibility of malignant transformation must be taken into account

Cocoa flavonoid-enriched diet modulates systemic and intestinal immunoglobulin synthesis in adult Lewis rats

Previous studies have reported that a diet containing 10% cocoa, a rich source of flavonoids, has immunomodulatory effects on rats and, among others effects, is able to attenuate the immunoglobulin (Ig) synthesis in both systemic and intestinal compartments. The purpose of the present study was focused on investigating whether these effects were attributed exclusively to the flavonoid content or to other compounds present in cocoa. To this end, eight-week-old Lewis rats were fed, for two weeks, either a standard diet or three isoenergetic diets containing increasing proportions of cocoa flavonoids from different sources: one with 0.2% polyphenols from conventional defatted cocoa, and two others with 0.4% and 0.8% polyphenols, respectively, from non-fermented cocoa. Diet intake and body weight were monitored and fecal samples were obtained throughout the study to determine fecal pH, IgA, bacteria proportions, and IgA-coated bacteria. Moreover, IgG and IgM concentrations in serum samples collected during the study were quantified. At the end of the dietary intervention no clear changes of serum IgG or IgM concentrations were quantified, showing few effects of cocoa polyphenol diets at the systemic level. However, in the intestine, all cocoa polyphenol-enriched diets attenuated the age-related increase of both fecal IgA and IgA-coated bacteria, as well as the proportion of bacteria in feces. As these effects were not dependent on the dose of polyphenol present in the diets, other compounds and/or the precise polyphenol composition present in cocoa raw material used for the diets could be key factors in this effect.

Determination of salivary glucose in healthy adults

Objectives: Our aim in this study was to determine the concentration of salivary glucose in healthy individuals and to compare it with the capillary glycemia. Study design: Samples of unstimulated whole saliva were collected from 63 non-diabetic patients. The concentration of salivary glucose and capillary blood was measured in all of the patients. The salivary glucose was determined by enzymatic method and spectrophotometry. The data was then analyzed using the Spearman correlation test, considering values of p<0.05 to be significant. Results: The whole sample consisted of 47.6% males and 52.4% women, with an average age of 37.5±15.7 years old. The average rates of unstimulated salivary flow were 0.41±0.21 ml/min among males and 0.31±0.15 ml/min among females. No significant difference was found based on these results (p=0.078). The average blood glucose among the males studied was 100.05±13.51 mg/dL, and among females, it was 99.5±13.9 mg/dL. The average salivary glucose for the whole sample was 5.97±1.87 mg/dL, with 5.91±2.19 mg/dL among males and 5.97±1.56 mg/dL among females, respectively, without presenting any significant differences (p=0.908). The concentration of salivary glucose did not present any statistically significant correlation with the capillary glycemia (p=0.732). Conclusions: The results suggest that the concentration of salivary glucose is not dependent on capillary glycemia and that the concentration of salivary glucose does not present significant differences between the measurements for males and females.

Salivary cotinine concentrations in daily smokers in Barcelona, Spain: a cross-sectional study

Background: Characterizing and comparing the determinant of cotinine concentrations in different populations should facilitate a better understanding of smoking patterns and addiction. This study describes and characterizes determinants of salivary cotinine concentration in a sample of Spanish adult daily smoker men and women. Methods: A cross-sectional study was carried out between March 2004 and December 2005 in a representative sample of 1245 people from the general population of Barcelona, Spain. A standard questionnaire was used to gather information on active tobacco smoking and passive exposure, and a saliva specimen was obtained to determine salivary cotinine concentration. Two hundred and eleven adult smokers (>16 years old) with complete data were included in the analysis. Determinants of cotinine concentrations were assessed using linear regression models. Results: Salivary cotinine concentration was associated with the reported number of cigarettes smoked in the previous 24 hours (R2 = 0.339; p < 0.05). The inclusion of a quadratic component for number of cigarettes smoked in the regression analyses resulted in an improvement of the fit (R2 = 0.386; p < 0.05). Cotinine concentration differed significantly by sex, with men having higher levels. Conclusion: This study shows that salivary cotinine concentration is significantly associated with the number of cigarettes smoked and sex, but not with other smoking-related variables.

Papillary cystoadenoma lymphomatosum (Warthin-like) of minor salivary glands

Papillary cystadenoma lymphomatosum is a benign salivary gland tumor most frequently located in the parotid gland (Warthin"s tumor). Its presentation in other major, or in minor, salivary glands is rare. Clinically, it manifests as a slow growing tumor, fluctuant on palpation due to its cystic morphology. The treatment of choice is complete excision with wide tumor-free margins. We present a 73-year-old female patient with an asymptomatic tumor of 8 years evolution in the right posterior area of the hard palate. We performed surgical excision and a biopsy, which was reported as papillary cystadenoma lymphomatosum. During the post-operative examination carried out after 3 weeks, it was observed that the lesion had recurred. The lesion was re-operated, performing the excision with CO2 laser and including the periosteum to ensure complete resection of the tumor. At 10 months follow-up, there was no recurrence of the lesion. This article includes a review of this condition and discusses its most important clinical and pathologic features and therapeutic approaches.

Epidermal growth factor secreted from submandibular salivary glands interferes with the lipolytic effect of adrenaline in mice

We had described that epidermal growth factor (EGF) interfered with the lipolytic effect of catecholamines in isolated adipocytes. Since catecholamines stimulate the release of EGF from submandibular salivary glands to blood plasma in male mice, we studied whether EGF affected also the lipolytic response to adrenaline in whole animals. We studied the effect of adrenaline in sialoadenectomized and sham-operated mice receiving or not a high dose of EGF following adrenaline injection. There was no difference in plasma EGF concentration between sham-operated and sialoadenectomized animals receiving saline. After adrenaline administration plasma EGF increased by 20-fold in sham-operated but did not increase in sialoadenectomized mice. Indeed, the increase was much higher (more than 100-fold) in mice receiving exogenous EGF. The effect of adrenaline on plasma concentration of both glycerol and nonesterified fatty acids was higher as lower was plasma EGF concentration. Isolated adipocytes obtained from sham-operated or sialoadenectomized mice had identical lipolytic response to adrenaline. The lipolytic response of adipocytes to isoproterenol was decreased by addition of EGF. To study whether the interference with the in vivo lipolytic effect of adrenaline had further metabolic consequences, we measured plasma b-hydroxybutyrate concentration in plasma. There was no difference in the response to adrenaline between sham-operated and sialoadenectomized mice in spite of the difference in plasma nonsterified fatty acid concentration. Studies in isolated hepatocytes indicated that ketogenesis run at near maximal rate in this range of substrate concentration. These results suggest that EGF in the physiological range decreases the lipolytic effect of adrenaline but does not compromise further metabolic events like the enhancement of ketogenesis.