Effect of partial dietary replacement of fishmeal by yellow mealworm (Tenebrio molitor) larvae meal on the innate immune response and intestinal antioxidant enzymes of rainbow trout (Oncorhynchus mykiss).

The present study investigated the effects of 25 and 50% dietary inclusion of yellow mealworm, Tenebrio molitor (TM), larvae on the intestinal antioxidant enzymes, lipid peroxidation and immune system of rainbow trout, Oncorhynchus mykiss. Fish were fed for 90 days with a control diet (TM0) or with 2 test diets (TM25 and TM50) where 35 and 67% of the fishmeal was replaced by TM larvae meal respectively. At the end of the growth trial, humoral immune parameters were assessed in the sera of the fish and the activities of antioxidant enzymes were measured in intestine homogenates. The present results show a strong increase of the activity of the antioxidant intestinal enzymes and reduction of lipid peroxidation. Increased trypsin inhibition and faster antibacterial activity of the serum were also detected. These evidences could be due to the physiological similarities between the exoskeleton of parasites and insects. The present findings reinforce the previous results which revealed a positive effect of the dietary TM on rainbow trout growth and further confirm its adequacy for fishmeal partial replacement in aquafeed.

Does dietary insect meal affect the fish immune system? The case of mealworm, Tenebrio molitor on European sea bass, Dicentrarchus labrax.

Feeding small European sea bass, Dicentrarchus labrax, for 6 weeks with Tenebrio molitor larval meal showed significant anti-inflammatory responses (ceruloplasmin, myeloperoxidase and nitric oxide). Serum bacteriolytic activity against a Gram negative bacterium was not significantly affected by dietary Tenebrio, while both lysozyme antibacterial activity and serum trypsin inhibition usually linked to the anti-parasite activity of the fish, were significantly enhanced. The latter may be due to the similarities in the composition of the exoskeleton of parasites and insects that may therefore act as an immunostimulant potentially increasing the anti-parasitic activity. The addition of exogenous proteases significantly decreased both trypsin-inhibition and serum bacteriolytic activity probably through direct inhibition of the proteins responsible for these immune functions. Further investigation involving bacterial or parasitic challenges will be necessary to assess if the effects of dietary mealworm meal on the immune system observed in the present study are translated into an improved resistance to diseases.

Mutational profile of TP53 in esophageal squamous cell carcinoma associated with chagasic megaesophagus.

Chaga's disease is an important communicable neglected disease that is gaining wider attention due to its increasing incidence worldwide. Achalasia due to chagasic megaesophagus (CM), a complication of this disease, is a known-yet, poorly understood-etiological factor for esophageal squamous cell carcinoma (ESCC) development. In this study, we aimed to perform the analysis of TP53 mutations in a series of Brazilian patients with ESCC that developed in the context CM (ESCC/CM), and to compare with the TP53 mutation profile of patients with benign CM and patients with nonchagasic ESCC. Additionally, we intended to correlate the TP53 mutation results with patient's clinical pathological features. By polymerase chain reaction (PCR) followed by direct sequencing of the hotspot regions of TP53 (exon 5 to 8), we found that TP53 mutations were present in 40.6% (13/32) of the ESCC/CM group, 45% (18/40) of the nonchagasic ESCC group, and in only 3% (1/33) of the benign CM group. Missense mutations were the most common in the three groups, yet, the type and mutated exon mutation varied significantly among the groups. Clinically, the groups exhibited distinct features, with both cancer groups (ESCC and ESCC/CM) been significantly associated higher consumption of alcohol and tobacco, older age, worse Karnofsky performance status, poor outcome than the patients with benign CM. No significant association was found between TP53 mutation profile and clinical-pathological features in any of the three groups. We describe first the time the analysis of TP53 mutations in ESCC that developed in the context of CM, and the observed high frequency of mutations, suggest that TP53 also plays an important role in the tumorigenic process of this unexplored etiological condition.

A Comparative, Prospective and Randomized Evaluation of Roux-en-Y Gastric Bypass With and Without the Silastic Ring: A 2-Year Follow Up Preliminary Report on Weight Loss and Quality of Life.

BACKGROUND: Currently, Roux-en-Y gastric bypass (RYGB) is one of the most widely used bariatric surgeries. Banding the pouch forms a banded gastric bypass operation, an accepted and frequently used variant. Placing a silastic ring around the pouch to band the gastric bypass operation increases the restriction mechanism. However, the ubiquitous use of the banded gastric bypass remains controversial. One of the controversies is the effect of the silastic ring on patients' perception of their well being after surgery because of the frequency of vomiting. A prospective, blindly randomized, comparative trial was undertaken to resolve this controversy. METHOD: Four hundred subjects scheduled for gastric bypass surgery were randomized into two arms of the trial, 200 with a silastic ring (WR) and 200 without (NR). After 2-year follow-up, the variables associated with the scores of Bariatric Analysis and Reporting Outcome System (BAROS) were analyzed. RESULTS: The initial median weight (125 kg), BMI (47), and age (36 years) were the same in both the NR and WR groups. The median excess weight loss, weight regain, and incidence of vomiting were 71, 10.5, and 7.75%, respectively, in the NR group vs. 75.4 and 1.1, and 24.4% in the WR group. The mean QOL score was 79% in the NR group vs. 80% in the WR group. CONCLUSION: After 2-year follow-up, silastic ring placement in the RYGB resulted in greater weight loss and weight stability and a threefold greater incidence of vomiting. There was no difference in the scores in the quality of life analysis.

Delivery of Apical Mesenchymal Stem Cells into Root Canals of Mature Teeth.

Regenerative endodontic procedures are stem cell-based treatments for immature teeth with pulp necrosis. The translation of regenerative endodontic procedures into treating mature teeth depends, among other factors, on the availability and delivery of mesenchymal stem cells (MSCs) into the root canal system. The aim of this clinical study was to evaluate whether evoked bleeding from the periapical tissues elicits the influx of MSCs into the root canal system in mature teeth with apical lesions. Participants included in this study (N = 20) were referred for endodontic treatment of mature teeth with apical lesions. Following chemomechanical debridement, intracanal bleeding from the periapical tissues was achieved, and intracanal blood samples were collected. A positive blood aspirate was also collected in the cartridges during local anesthesia. Total RNA was isolated and used as a template in quantitative reverse transcription polymerase chain reactions using MSC-specific arrays. Data were analyzed with the Wilcoxon signed-rank test, and correlation between gene expression and sex or age was tested with Spearman's rank correlation coefficient test. In addition, MSCs were isolated from an intracanal bleeding sample and subjected to flow cytometry and quantitative osteogenesis assay. Last, the presence and distribution of MSCs within periradicular lesions were evaluated with immunohistochemistry (n = 4). The MSC markers CD73, CD90, CD105, and CD146 were significantly upregulated, with median fold change values of 2.9, 31.7, 4.6, and 6.8, respectively. Conversely, the negative marker for MSCs, CD45, was significantly downregulated (median, -2.7). There was no correlation with age, sex, tooth type, or treatment for any of the evaluated genes. Isolated intracanal cells coexpressed MSC markers and demonstrated robust mineralizing differentiation potential. Finally, immunohistochemical analysis revealed that MSCs were found compartmentalized mainly within vasculature structures located in periapical lesions. Collectively, findings indicate that the evoked-bleeding technique delivers MSCs into the root canal system in mature teeth with apical lesions.

Strong effect of long-term Sparicotyle chrysophrii infection on the cellular and innate immune responses of gilthead sea bream, Sparus aurata.

One thousand healthy recipient gilthead sea bream, Sparus aurata, cohabited with 250 donor fish parasitized by Sparicotyle chrysophrii (Van Beneden and Hesse, 1963) (Monogenea: Polyopisthocotylea), a common parasite of the gills of this fish species. Controls consisted of 1000 healthy fish kept in a separate tank. After 10 weeks, fish were weighed and parasite load, hemoglobin concentration and immunological parameters were assessed. Rather than the absence of parasite, hemoglobin concentration was a better marker of the health status of the fish, because S.chrysophrii had detached from the strongly anemic gills of some animals leaving fish with affected immune system but without parasites. The parasite infection seemed to trigger a cellular response of the fish immune system but to inhibit its humoral components. Thus, parasitized fish may control the parasite infection through the action of reactive oxygen species but they may become more sensitive to potential secondary bacterial or parasitical infections. This phenomenon was demonstrated not only through significant differences between recipient and control fish but also through strong correlations between those parameters and parasite load, fish weight and/or hemoglobin concentration.

Sex-dependent roles of prolactin and prolactin receptor in postoperative pain and hyperalgesia in mice.

Although surgical trauma activates the anterior pituitary gland and elicits an increase in prolactin (PRL) serum levels that can modulate nociceptive responses, the role of PRL and the PRL-receptor (PRL-R) in thermal and mechanical hyperalgesia in postoperative pain is unknown. Acute postoperative pain condition was generated with the use of the hindpaw plantar incision model. Results showed endogenous PRL levels were significantly increased in serum, operated hindpaw and spinal cords of male and female rats 24h after incision. These alterations were especially pronounced in females. We then examined the role of the PRL system in thermal and mechanical hyperalgesia in male and female mice 3-168 h after plantar incision with the use of knock-out (KO) mice with PRL or PRL-R gene ablations and in wild-type (WT) mice. WT mice showed postoperative cold hyperalgesia in a sex-dependent manner (only in females), but with no effect on heat hyperalgesia or mechanical allodynia in either sex. Studies in KO mice showed no effect of PRL and PRL-R gene ablation on heat and cold hyperalgesia in male mice, while heat hyperlgesia were reduced 3-72 h post-surgery in female PRL and PRL-R KO mice. In contrast, PRL and PRL-R ablations significantly attenuated mechanical allodynia 3-72 h post-surgery in both male and female mice. Overall, we found elevated PRL levels in serum, hindpaws and spinal cords after incision, and identify a contributory role for the PRL system in postoperative pain responses to thermal stimuli in females and to mechanical stimuli in both males and females.

In vitro effect of peas, Pisum pisum, and chickpeas, Cicer arietinum, on the immune system of gilthead seabream, Sparus aurata.

The future for a sustainable aquaculture relies on the formulation of feed including alternatives to fish meal and fish oil that do not impair fish growth and that improve fish health status. Grain legumes such as field peas and chickpeas offer good sources of proteins, carbohydrates, fibers, vitamins, and minerals. The effect of peas and chickpeas on the immune system of seabream was assessed in vitro in order to detect any potential immunosuppressing problem. Peas was determined to be a better fishmeal alternative than chickpeas as they induced higher respiratory burst measured by the nitro blue tetrazolium assay and primed the Phorbol 12-myristate 13-acetate (PMA)-stimulated intracellular respiratory burst whereas chickpeas neither directly stimulated respiratory burst nor primed it. However, when the intra- and extracellular respiratory burst activities were taken into account, high concentrations of peas inhibited the zymosan- and PMA-triggered chemiluminescence. This apparent reduction of the production of reactive oxygen species may reflect in fact the antioxidant activity of legumes. This, together with the absence of effect on the phagocytosis activity, suggested that peas are not immunosuppressing gilthead seabream. Further in vivo studies preferably comporting a bacterial challenge will have to ascertain the absence of immunosuppressing effect of these legumes.

LPS sensitizes TRPV1 via activation of TLR4 in trigeminal sensory neurons.

Recent studies have demonstrated that the lipopolysaccharide (LPS) receptor (TLR4) is expressed in TRPV1 containing trigeminal sensory neurons. In this study, we evaluated whether LPS activates trigeminal neurons, and sensitizes TRPV1 responses via TLR4. To test this novel hypothesis, we first demonstrated that LPS binds to receptors in trigeminal neurons using competitive binding. Second, we demonstrated that LPS evoked a concentration-dependent increase in intracellular calcium accumulation (Ca(2+))(i) and inward currents. Third, LPS significantly sensitized TRPV1 to capsaicin measured by (Ca(2+))(i), release of calcitonin gene-related peptide, and inward currents. Importantly, a selective TLR4 antagonist blocked these effects. Analysis of these data, collectively, demonstrates that LPS is capable of directly activating trigeminal neurons, and sensitizing TRPV1 via a TLR4-mediated mechanism. These findings are consistent with the hypothesis that trigeminal neurons are capable of detecting pathogenic bacterial components leading to sensitization of TRPV1, possibly contributing to the inflammatory pain often observed in bacterial infections.

Pulpitis increases the proportion of atypical nodes of Ranvier in human dental pulp axons without a change in Nav1.6 sodium channel expression.

Studies show a change in sodium channel (NaCh) expression after inflammatory lesions, and this change is implicated in the generation of pain states. We are using the extracted human tooth to study NaCh expression and here examine the expression of the major NaCh isoform located at nodes of Ranvier, Na(v)1.6, in normal and painful samples. Pulpal sections were double-labeled with human-specific Na(v)1.6 antibody and caspr antibody (paranodal protein to identify nodes). Confocal microscopy was used to obtain a z-series of optically-sectioned images of axon bundles surrounded by inflammatory cells in painful samples and of similar regions within the coronal pulp of normal samples. Nodes contained within these images were classified as typical or atypical as based on caspr staining relationships, and NIH ImageJ software was used to quantify the size and immunofluorescence staining intensity of Na(v)1.6 accumulations at these nodal sites. Results show no significant difference in the size or immunofluorescence staining intensity of Na(v)1.6 nodal accumulations located at either typical or atypical nodal sites (heminodes and split nodes) within axons in normal samples when compared to painful samples (n=9/each group). In contrast, there was a highly significant decrease in the proportion of typical nodal sites and an increase in atypical nodal sites in painful samples when compared to normal samples. The unchanged expression of Na(v)1.6 contrasts to our previous finding that showed an increased expression of Na(v)1.7 at both typical and atypical nodal sites within painful samples. Together, these findings suggest there is not a simple replacement of one isoform with another, but rather an increased co-expression of multiple isoforms at both intact and remodeling/demyelinating (atypical) nodal sites within the painful dental pulp. The resultant heterogeneous population of isoforms may produce unique axonal excitability properties that could contribute to spontaneous pain sensations that are common in toothache.

Effects of a natural parasitical infection (Lernanthropus kroyeri) on the immune system of European sea bass, Dicentrarchus labrax L.

The immune response of European sea bass, Dicentrarchus labrax, to a natural infection by the copepod parasite Lernanthropus kroyeri was evaluated for the first time in vivo. The results clearly demonstrated the triggering of the fish immune system by the parasite. Lysozyme activity and alternative complement pathway were involved in the early action against the parasitical infection, whilst classical complement and respiratory burst (RB) activity took over in the later stages of infection. It was hypothesized that the levels of alternative and classical complement and RB stimulation indexes may determine the resistance capacity of the fish to the parasite. It is not clear how parasites can survive despite the strong immunological arsenal deployed by the fish. The continual increase of prevalence and severity of parasite infection suggested that the parasite's mechanism of evasion of the immune system was extremely successful. The contrasting decrease in the negative effects of parasites on the fish health (such as gills anaemia) suggested that an equilibrium between the parasites and their hosts was reached in chronic infection. These dynamic interactions between parasites and fish hosts were probably the main determinant of host specificity.

Tumor necrosis factor alpha enhances the sensitivity of rat trigeminal neurons to capsaicin.

Tumor necrosis factor alpha (TNFalpha), a pro-inflammatory cytokine, enhances the development of pain and hyperalgesia, although the molecular mechanisms are not well understood. This study evaluated the hypothesis that TNFalpha increases the sensitivity of rat trigeminal neurons to capsaicin via two different mechanisms triggered by either brief or sustained exposure to the cytokine. A brief (5 min) application of TNFalpha significantly sensitized capsaicin-evoked accumulation of intracellular calcium ([Ca2+]i) (226.4+/-37.7 nM vs. 167.5+/-31.3 nM) and increased capsaicin-evoked nocifensive behavior (78.3+/-9.7 vs. 30.9+/-3.6 s) as compared with vehicle pretreatment (P<0.01 for both). Sustained (30 min to 4 h) exposure of cultured neurons to TNFalpha evoked a twofold increase in mRNA transcript (P<0.05) and protein levels (P<0.01) of transient potential receptor vanilloid type 1 (TRPV1). This long-term up-regulation of TRPV1 expression by TNFalpha correlated with enhancement in capsaicin-induced calcitonin gene-related peptide release (P<0.05). Demonstration of colocalization of TNFalpha receptor subtypes I and II with TRPV1 in almost all (>90%) TRPV1 expressing neurons provides evidence consistent with a direct interaction on the same subpopulation of sensory neurons. In summary, our data demonstrate that TNFalpha directly enhances the sensitivity of rat trigeminal neurons to capsaicin via both rapid, non-genomic mechanisms as well as sustained genomic regulation in TRPV1 expression. Thus, increased sensitization and up-regulation of TRPV1 constitutes a potential mechanism by which TNFalpha mediates inflammatory hyperalgesia and pain.

[Electromanometry of the upper esophageal sphincter before and after esophageal perfusion with hydrochloric acid 0,1N. Experimental study in dog]. / Eletromanometria do esfíncter superior do esôfago antes e após perfusão esofágica com ácido clorídrico 0,1N. Estudo experimental no cão.

BACKGROUND: The responses of the upper esophageal sphincter to gastroesophageal reflux is controversial. OBJECTIVE: Study the effect of upper esophageal sphincter to the esophageal acid perfusion. METHODS: Thirty adult dogs of both sexes were studied, being submitted to esophageal electromanometry. The pull through technique and continuous infusion of the catheters with distilled water were employed. These exams allowed us to measure the pressure width (mm Hg) and the length (cm) of the upper esophageal sphincter in basal conditions (moment 1). After this first phase the animals were submitted to esophagic infusion, being then divided in three groups, according to the solution used in the infusion and the moment of the study, as follows: Group 1: esophagic infusion with distilled water and electromanometric studies accomplished 15 minutes (moment 2) and 30 minutes (moment 3) of the end of the infusion. Group 2: esophagic infusion with HCl 0.1 N and electromanometric studies accomplished 15 minutes after the end of the infusion (moment 2). Group 3: esophagic infusion with HCl 0.1 N and electromanometric studies accomplished 30 minutes after the end of the infusion (moment 3). RESULTS/CONCLUSIONS: This research was performed to evaluate the esophagic acidification influence on the upper esophageal sphincter. The observed results allowed us to conclude that the acidification of the esophagus did not cause any alteration on the pressure width and on the length of the upper esophageal sphincter.

Functional evaluation of the ileocolic sphincter transplanted to the neck using microsurgery technique: a manometric study in dogs.

The usefulness of ileocolic sphincter (ICS) transplantation to the neck after a total laryngopharyngectomy was investigated in 12 mongrel dogs. The authors aimed to evaluate the influence of the transplantation on ICS pressure amplitude and ICS length, and on ICS resistance to intracecal pressure increase. Electromanometric measurements showed that ICS pressure amplitude and length presented no changes after the procedure. Also, there was no significant change in ICS resistance. However, long-term follow-up studies are needed to determine whether the ileocolic transition maintains the properties observed in the immediate postoperative period.

Esophageal motor disturbances in progressive systemic sclerosis.

Ten patients with progressive systemic sclerosis (PSS) and esophageal symptoms (group 1) and 10 control subjects were studied. Esophageal electromanometry using the intermittent pull-through technique and catheter perfusion with distilled water were performed in all patients and individuals. The variables studied were pressure amplitude in the lower esophageal sphincter (LES) (mmHg) and deglutition wave amplitude (mmHg at 5, 10 and 15 cm above LES). In PSS patients, the average LES pressure was 18.5 +/- 4.6 mmHg, and in control subjects it was 27 +/- 6.5 mmHg (p < 0.01). Deglutition wave amplitude at 5, 10 and 15 cm above LES was 13.2 +/- 7.5 mmHg, 12 +/- 3.7 mmHg and 15 +/- 3.3 mmHg, respectively, in PSS patients. In control subjects, it was 67.6 +/- 12.5 mmHg, 58.6 +/- 20.9 mmHg and 52.4 +/- 21.4 mmHg (p < 0.001). In PSS patients, the pressure amplitude in LES and in the body of the esophagus was lower than in control subjects. In PSS patients, esophageal manometry showed the absence of normal peristalsis.

[Use of botulin toxin in the treatment of diffuse esophageal spasm. A case report]. / Emprego da toxina botulínica no tratamento do espasmo difuso esofagiano. Relato de caso.

The diffuse esophageal spasm distinguishes from others esophageal disturbs by the symptoms severity and bad results to conservative treatment. This situation requires formal indication to surgical treatment or dilation. In high risk patients this situation becomes a big problem, due to the lack of another therapeutic option. In a patient with this problem we use an endoscopic injection of botulin toxin in cardia with good results in three months follow-up. Despite this short period of observation we believe that the infiltration with botulin toxin in cardia constitutes a valid therapeutic option in treatment of diffuse esophageal spasm.

Oocyte maturation in rabbits: effects of calmodulin inhibitors.

Oocyte maturation in mammals follows a highly conserved pattern of release from arrest through to the extrusion of the first polar body and formation of the second metaphase spindle. Oscillations in cytoplasmic calcium concentration precede the events of maturation in many species. These calcium ions interact with and activate calcium-binding proteins, including calmodulin, within the cell. Thus, it was of interest to us to examine whether calcium acted through calmodulin in the initial stages of maturation in rabbit oocytes or whether calmodulin was required for continuation through metaphase I no to metaphase II. Using the calmodulin inhibitor W-7 we found a significant (p < 0.05) decrease in the percentage of oocytes that underwent germinal vesicle breakdown. Calmidazolium did not prevent germinal vesicle breakdown; however, it caused a significant (p < 0.05) decrease in the proportion of oocytes with fully elaborated spindles and taxol-induced cytoplasmic asters. Both inhibitors caused a significant (p < 0.05) reduction in the proportion of oocytes that extruded their first polar bodies. The kinase inhibitor 6-DMAP caused a significant reduction in the proportion of oocytes with spindles and condensed chromatin, indicating the necessity for phosphorylation events in the resumption of meiosis. In rabbit oocytes calmodulin may play a role in the release from prophase arrest, and it is necessary for spindle preservation and continuation through metaphase I to metaphase II. The varying effects of the two inhibitor stems from their different binding sites on the calmodulin molecule thus causing a differential effect on its downstream effectors.

Laryngeal mobility and dysphagia.

The authors studied the utility of the physical test of laryngeal movement in swallowing disorders of the oropharyngeal region. Measurement and palpation of the larynx during deglutition were performed in the neck of 14 dysphagic patients and in two normal control groups. The normal groups were used to establish the pattern of the movement and the normal values of laryngeal elevation. Control elevation ranged from 1.80 to 2.50 cm. In eight patients laryngeal motion was defective and presented values ranging from zero to 1.50 cm. Palpation during laryngeal movement also revealed unexpected anomalous displacement such as lateral shifting and lowering of the larynx. In six patients with defective laryngeal motion, pharyngeal and upper esophageal sphincter function were also impaired. Direct measurement and palpation of laryngeal mobility during deglutition is a noninvasive method that can be used to evaluate dysphagia and the risk for aspiration. Also, it allows physical assessment of the evolution of the disorder.