Infant twin mortality and hospitalisations after the perinatal period - a prospective cohort study from Guinea-Bissau.

OBJECTIVE: To examine mortality and hospitalisations among infant twins and singletons after the perinatal period in Guinea-Bissau. METHODS: The study was conducted from September 2009 to November 2012 by the Bandim Health Project (BHP). Newborn twins and unmatched singleton controls were included at the National Hospital Simão Mendes in the capital Bissau. Children were examined clinically at enrolment. Maternal, pregnancy and obstetric information was collected and HIV testing offered at birth. Follow-up occurred at home at 2, 6 and 12 months and through linkage with the paediatric admission register at the National Hospital. RESULTS: About 495 twins and 333 singletons were alive on day 7 after birth. In total, 36 twins and 12 singletons died during follow-up, the post-perinatal infant mortality rate being 91/1000 person-years for twins and 42/1000 for singletons (HR = 2.11, 95% CI: 1.09-4.07). In a multivariable analysis among twins only, birth weight <2000 g [3.32, (1.36-8.07)], death of the cotwin perinatally [2.54, (1.16-5.57)] and severe maternal illness during pregnancy [2.35, (1.00-5.51)] were significant risk factors for twin death. In the subgroup with available HIV status, maternal HIV infection was strongly associated with twin mortality [3.16, (1.24-8.05)]. Death occurred at home for 60% of twins and 67% of singletons. During follow-up, 90 first-time hospital admissions were registered, with similar rates observed for twins (139/1000) and singletons (143/1000) [0.97, (0.61-1.52)]. CONCLUSION: The post-perinatal infant mortality rate of twins was double that of singletons. No excess in twin hospitalisations was observed, possibly implying obstacles to hospital admission for twins in case of severe illness.

Evaluation of Bond Strength and Fracture Load of Teeth With Flared Root Canals Restored With Prefabricated, Anatomical, and Computer-aided Design and Computer-aided Manufacturing Fiber Posts.

PURPOSE: This study compared the use of anatomical glass fiber posts using bulk-fill composite resin with computer-aided design and computer-aided manufacturing (CAD/CAM) milled glass fiber post in oversized root canals, through bond strength (BS) and fracture resistance (FR) tests (fracture load). METHODS AND MATERIALS: A total of 48 mandibular premolars were selected, half of them had their crowns removed at the cemento-enamel junction (CEJ) and the other half were sectioned 2 mm above the CEJ. Subsequently, teeth were endodontically treated. After 1 week, the standard preparation of the canals was carried out, and the roots were divided into three groups (n=16), according to the use of different restorative techniques (control: prefabricated glass fiber post [PFP], direct anatomical glass fiber post [AFP], and CAD/CAM milled glass fiber post [MFP]). After luting procedures using Single Bond Universal and RelyX Ultimate (3M ESPE), for eight teeth in each group, six specimens were obtained (two slices from each root third: cervical, middle, and apical). For the remaining eight roots of each group, standardized preparations for metal-free crowns, milling of 5 mol% yttria-stabilized tetragonal zirconia polycrystalline, cementation of the crowns, and periodontal ligament simulation were performed. Then, for each group, the BS was evaluated through the push-out test, and the FR was evaluated in compression. The data obtained from BS were submitted to two-way analysis of variance (ANOVA; group vs root region) and Tukey (α=0.05) and from FR to one-way ANOVA (group) and Tukey (α=0.05). RESULTS: For the BS test, the MFP group was statistically superior to the PFP group in all root regions and was statistically superior to the AFP group only in the cervical region, being statistically similar in the middle and apical root regions. For the FR test, the MFP group was statistically superior to the PFP and AFP groups. CONCLUSION: The milled fiber post technique can be a legitimate alternative in the restoration of weakened roots with flared root canals.

Immunohistochemical staining of human islet cells with region-specific antibodies against secretogranins II and III.

Chromogranins and secretogranins belong to the granin family of proteins, which are expressed in neuroendocrine and nervous tissue. In earlier publications we have described the development of region-specific antibodies against CgA and CgB. In this study we describe antibodies to SgII and SgIII and their usefulness for immunohistochemical staining. Peptides homologous to defined parts of secretogranins II and III were selected and synthesized. Antibodies were raised and immunostainings were performed on normal human pancreas. The SgII 154-165 (N-terminal secretoneurin), SgII 172-186 (C-terminal secretoneurin) and SgIII antibodies immunostained all insulin-immunoreactive cells, most of the glucagon cells and some of the pancreatic polypeptide cells. The SgII 225-242 antibody immunostained only the insulin-containing cells. None of the antibodies immunostained the somatostatin cells. This study is the first observation of the expression of SgIII in human tissues, where we show expression of SgIII in three of the four major islet cell types in human pancreas.

The importance of chromogranin A in the development and function of endocrine pancreas.

BACKGROUND: Chromogranin (Cg) A is expressed in neuroendocrine and neuronal tissues. It is involved in the generation of secretory granules and is cleaved to form biologically active peptides. Targeted ablation of the Chga gene resulted in increased plasma catecholamines, high blood pressure, and decreased size and number of adrenal medullary chromaffin granules. The aim of this study was to determine whether Chga null mice display changes in the morphology and function of the endocrine pancreas. MATERIALS AND METHODS: Sections of pancreata from Chga-/-, Chga+/- and Chga+/+ mice, were immunostained with antibodies against synaptophysin, CgA, CgB, secretogranin II and the four major pancreatic islet hormones. Plasma was analysed for glucose, insulin, glucagon, somatostatin and pancreatic polypeptide (PP). RESULTS: CgA epitopes were undetectable in the islets of Chga-/- animals. CgB and secretogranin II epitopes were expressed in the islets of all animal groups albeit with decreased expression in Chga-/- islets. The islet number and size were decreased in the Chga-/- animals compared with Chga+/+. The proportion of insulin cells was decreased but somatostatin and PP cells were increased in Chga-/- mice compared to Chga+/+ mice. The nuclear size was decreased in insulin cells and increased in somatostatin cells in Chga-/- mice. Plasma insulin level was markedly decreased in the Chga-/- mice although fasting plasma glucose and glucagon were normal. CONCLUSION: Ablation of the Chga gene affected the islet volume, the composition, distribution and nuclear size of islet cell types and plasma insulin concentration. Our data indicate decreased insulin cell function and increased glucagon cell function. Our study shows that CgA exerts a significant influence on the endocrine pancreas with importance in maintaining islet volume, cellular composition and function.

Efficacy of the liposome trifluralin in the treatment of experimental canine leishmaniosis.

Liposomes are used as carriers to deliver drugs and to treat diseases where infection is localised in the mononuclear phagocyte system cells, as is the case of leishmaniosis. Trifluralin is a dinitroaniline with proved anti-Leishmania activity in vitro. The efficacy of liposomal trifluralin (LIP/TFL) was studied in the treatment of experimental canine leishmaniosis through quantification of parasite burden using the limiting dilution assay, follow-up of anti-Leishmania antibodies by indirect fluorescent immunoassay and cytokine expression by Reverse Transcriptase-PCR, in the bone marrow, lymph nodes, skin and peripheral blood mononuclear cells in 5 female beagle dogs. After treatment, dogs showed a general remission of clinical signs related to parasite burden reduction and Th1 cytokine mRNA expression, but there was no significant decrease in antibody levels. Alternative treatment schemes with LIP/TFL are necessary to achieve optimal results.

Three-Year Effects of Deproteinization on the In Vitro Durability of Resin/Dentin-Eroded Interfaces.

OBJECTIVE: To evaluate the effect of sodium hypochlorite on the immediate and three-year bonding properties of a resin-eroded dentin interface produced by one of two adhesive strategies. METHODS AND MATERIALS: Forty-eight molars were randomly assigned to six experimental groups, according to the combination of the adhesive strategy (etch-and-rinse and self-etch) and the dentin surface (control groups without erosion, eroded dentin surface [ED], and eroded dentin surface + NaOCl 5.2% [ED + NaOCl]). After completing restoration, specimens were stored in water (37°C) for 24 hours and then sectioned into resin-dentin beams (0.8 mm2) to be tested under tension (0.5 mm/min) immediately thereafter or after three years of water storage. To assess nanoleakage (NL), specimens were immersed in silver nitrate solution and examined by scanning electron microscopy at both time points. The dentin-etching pattern was examined under a scanning electron microscope. Data were subjected to appropriate statistical analysis (α=0.05) Results: In both strategies, a more pronounced and significant reduction of the microtensile bond strength (µTBS) values was observed for the ED groups ( p=0.0001) after three years. However, in the ED + NaOCl group, µTBS values were maintained after three years of water storage. Furthermore, application of NaOCl to eroded dentin significantly reduced the immediate NL values and also preserved these values after three years of water storage for both adhesive strategies ( p>0.05). When considering the ED group, a superficial removal of the smear layer and enlarged lumen tubules in comparison to control were present. However, for ED + NaOCl, there was a total removal of the smear layer and significant numbers of collagen fibrils were exposed. CONCLUSION: The use of NaOCl may maintain the long-term stability of a resin-eroded dentin interface formed by etch-and-rinse and self-etch adhesives.

Using CAD/CAM-Modified Correlation Mode to Produce Laminate Veneers: A Six-Month Case Report.

The expectation of an esthetically harmonious smile increases the level of difficulty when treating patients. Laminate veneers stand out as a treatment option for cosmetic rehabilitation in clinical practice, as they are a more conservative procedure and mimic dental structures. These laminate veneers are generally made with different techniques; the most common requires an impression of the prepared tooth, an impression antagonist, fabrication models, and extensive laboratory time. The computer-aided design/computer-aided manufacturing (CAD/CAM) system optimizes the fabrication of prosthetic structures, reducing chairside time and promoting good esthetic results. Thus, the purpose of this case report is to present the esthetic result of multiple CAD/CAM manufactured laminate veneers using a new self-etching glass ceramic primer with a lithium disilicate ceramic, using the modified correlation and biogeneric modes.

Four-year Follow-up of a Direct Anatomical Fiber Post and Esthetic Procedures: A Case Report.

Knowledge about the stability of fiber posts cemented in widened canal spaces over time is scarce in the literature. Thus, the purpose of this case report was to evaluate the performance of a direct anatomical post in a widened canal space over the course of four years. The present clinical case describes the rehabilitation of a widened canal space using a direct anatomical post (a resin composite combined with a prefabricated glass fiber post) associated with an all-ceramic crown and other restorative procedures. This technique is easy to perform and may solve some of the problems associated with the cementation of a poorly adapted fiber post in a widened canal space.

Use of a Direct Anatomic Post in a Flared Root Canal: A Three-year Follow-up.

The following case report describes the three-year follow-up after rehabilitation of a flared root canal using a direct anatomic post (a resin composite combined with a prefabricated glass fiber post) associated with metal-free ceramic restoration. The report presents the clinical protocol for the fabrication of the posts, which provide an intimate fit to the remaining root and mechanical properties similar to those of the dental structure. These posts serve as an alternative to conventional metal cores.

Chromogranin A in human neuroendocrine tumors: an immunohistochemical study with region-specific antibodies.

Antibodies to six specific regions of the chromogranin A (CgA) molecule were used to study their immunoreactivity in human neuroendocrine (NE) tumors. Tissue specimens from endocrine pancreatic tumors (n = 14), duodenal carcinoids (n = 2), bronchial carcinoids (n = 5), ileal carcinoids (n = 5) appendix carcinoids (n = 2), medullary thyroid carcinomas (n = 6), parathyroid adenomas (n = 2), and pheochromocytomas (n = 8) were analyzed. The results showed that the NE tumor types expressed varying numbers of CgA fragments. A variation in frequency of the expression of immunoreactive cells was sometimes seen also within the same tumor type. The midportion fragment CgA 176-195 (chromacin) was the only fragment expressed in all tumors. Benign and malignant tumors expressed different patterns, being especially true of insulinomas and pheochromocytomas. These findings suggest that region-specific antibodies to CgA fragments can be used as a diagnostic tool for the characterization of NE tumors.

Selective processing of chromogranin A in the different islet cells in human pancreas.

We studied the immunoreactivity of 12 different region-specific antibodies to the chromogranin A (CgA) molecule in the four major neuroendocrine cell types of the human pancreas by using double immunofluorescence techniques. The antibodies raised to the N-terminal and midportions of CgA showed, on the whole, stronger immunoreactivity than did the C-terminal antibodies, with a few exceptions. Often the immunoreactivity was stronger in glucagon cells. Insulin cells expressed immunoreactivity to all region-specific antibodies, but glucagon cells were nonreactive to two antibodies. Somatostatin cells reacted only with the C-terminal antibodies (amino acid sequences CgA 411-424), while PP cells were stained with four CgA region-specific antibodies between amino acid sequences 63-195. The cause of these differences may be that the CgA molecule is cleaved, partly masked, or partly translated from CgA mRNA. Microwave treatment improved only the staining with the CgA 361-372 antibodies, which indicates that masking is not the sole or entire cause. Our findings may indicate that the CgA molecule is cleaved in different ways in the various pancreatic endocrine cell types, giving rise to a variety of biologically functional fragments.

Complex co-localization of chromogranins and neurohormones in the human gastrointestinal tract.

Co-localization of chromogranin (Cg) A, B, and C has been studied in different neuroendocrine cell types in histologically normal mucosa from human gastrointestinal tract (corpus, antrum, duodenum, ileum, and colon) using single-, double-, and triple-immunofluorescence stainings. Virtually all enterochromaffin (EC) cells contained CgA, and those in the luminal two thirds of the antral mucosa and villi of small intestine often also contained CgB. A few EC cells in the duodenal crypts contained CgC. Most gastrin cells harbored both CgB and CgA, although rather more CgB than CgA, but some gastrin cells contained all three types, i.e., also CgC. Some CCK cells also contained all three chromogranins. Enteroglucagon cells in the duodenal villi contained CgA and some CgB. CgA (but not B or C) was found in some secretin, GIP, enteroglucagon/peptide YY, and neurotensin cells. A few somatostatin cells contained CgA but neither CgB nor CgC. CgA and C were found mainly in the basal cell region, whereas CgB occurred more diffusely throughout the cytoplasm. This varying distribution suggests that not all secretory granules contain CgA, or that CgB may occur in a nongranular form. The varying composition of the different chromogranins may reflect their complex functional roles in the widespread neuroendocrine system.

Insulin-like growth factor II in human fetal pancreas and its co-localization with the major islet hormones: comparison with adult pancreas.

Insulin-like growth factor II (IGF-II) appears to play an important role during fetal life in cell growth and differentiation in several organs, including the pancreas. In the present study we investigated the cellular localization of IGF-II in human fetal pancreas at 16, 18 and 22 embryonic weeks and compared it with adult pancreas. Single and double immunofluorescence methods were used to study co-localization of IGF-II with the four major islet hormones - insulin, glucagon, somatostatin, pancreatic polypeptide - and with islet amyloid polypeptide (IAPP). Distinct IGF-II immunoreactive (IR) cells were found in the endocrine, but not in the exocrine, pancreas. The intensity of IGF-II immunoreactivity was more pronounced in the fetal than in the adult pancreas. In fetal pancreas IGF-II immunoreactivity was observed in virtually all insulin-IR cells and in subsets of the glucagon, somatostatin and IAPP cells. In the adult pancreas, IGF-II immunoreactivity was found in insulin/IAPP cells only. Our results suggest a broader effect of IGF-II in fetal endocrine pancreatic cells than in the adult.

Co-localization of neuroendocrine hormones in the human fetal pancreas.

OBJECTIVE AND DESIGN: Co-localization of the four major pancreatic hormones, and also of islet amyloid polypeptide (IAPP), peptide tyrosine tyrosine (PYY), secretin and neurotensin, has been studied in the endocrine pancreas of human fetuses at 16, 18 and 22 weeks of gestation. METHODS: Double and triple immunofluorescence stainings have been used. RESULTS: All three fetal pancreata contained cells that showed insulin, glucagon, somatostatin, pancreatic polypeptide (PP), IAPP, secretin and PYY immunoreactivity. Neurotensin cells were found in the youngest fetus and gastric inhibitory polypeptide (GIP) in the two older fetuses. Co-localization of two hormones occurred in most of the endocrine cell types in the three fetuses examined, but three hormones occurred in only a few cells and especially in the youngest fetus. Somatostatin cells were the only cell type which was largely monohormonal. Our findings showed that there are two different co-localization patterns: insulin was co-localized mainly with IAPP and glucagon, while secretin and PYY occurred together with glucagon and PP. CONCLUSIONS: These data are the first to describe secretin and neurotensin in the fetal pancreas. Two different co-localization patterns could be distinguished: insulin, IAPP and glucagon, and glucagon, secretin, PP and PYY.

Serum gastrin and gastrin-immunoreactive cells in the antral mucosa of patients with alcoholic liver disease.

BACKGROUND/AIMS: Hypergastrinaemia has been reported in liver cirrhosis; meanwhile, it is unclear whether it is associated with an increase in gastrin cell function. The serum gastrin concentration and the number of gastrin cells in antral biopsies were studied in patients with alcoholic liver disease. METHODS: Immunocytochemical and quantification techniques were used to localize and determine the number of gastrin cells. RESULTS: Slight non-significantly higher serum gastrin values were observed in the alcoholic liver disease patients compared with controls, but the individual variation within the groups was considerable. The frequency of gastrin cells did not differ between groups. However, the size of the gastrin cell nuclei was larger in patients with liver disease than in controls, indicating increased cellular activity. CONCLUSIONS: Alcoholic liver disease, with a disturbed liver function, influences the gastrin cells. The observed alterations may reflect the effect of alcohol and/or malnutrition, or may be secondary to the influence of liver disease on other regulatory peptides.

Isolation of Paracoccidioides brasiliensis from armadillos (Dasypus noveminctus) captured in an endemic area of paracoccidioidomycosis.

Paracoccidioides brasiliensis, the causative agent of paracoccidioidomycosis (PCM), was first isolated from armadillos from the Amazonian region where the mycosis is uncommon. In the present study, we report on the high incidence of PCM infection in armadillos from a hyperendemic region of the disease. Four nine-banded armadillos (Dasypus novemcinctus) were captured in the endemic area of Botucatu, Sao Paulo, Brazil, killed by manual cervical dislocation and autopsied under sterile conditions. Fragments of lung, spleen, liver, and mesenteric lymph nodes were processed for histology, cultured on Mycosel agar at 37 degrees C, and homogenized for inoculation into the testis and peritoneum of hamsters. The animals were killed from week 6 to week 20 postinoculation and fragments of liver, lung, spleen, testis, and lymph nodes were cultured on brain heart infusion agar at 37 degrees C. Paracoccidioides brasiliensis was isolated from three armadillos both by direct organ culture and from the liver, spleen, lung, and mesenteric lymph nodes of hamsters. In addition, one positive armadillo presented histologically proven PCM disease in a mesenteric lymph node. The three armadillos isolates (Pb-A1, Pb-A2, and Pb-A4) presented thermodependent dimorphism, urease activity, and casein assimilation, showed amplification of the gp43 gene, and were highly virulent in intratesticularly inoculated hamsters. The isolates expressed the gp43 glycoprotein, the immunodominant antigen of the fungus, and reacted with a pool of sera from PCM patients. Taken together, the present data confirm that armadillos are a natural reservoir of P. brasiliensis and demonstrate that the animal is a sylvan host to the fungus.

Expression of the five different somatostatin receptor subtypes in endocrine cells of the pancreas.

Knowledge concerning tissue-specific expression of the five somatostatin receptor subtypes is of great importance in understanding their physiological function. We developed rabbit polyclonal antibodies specific for each human somatostatin receptor subtype and report our results concerning the expression in normal endocrine pancreatic cells. The antibodies were produced by immunizing rabbits with fragments specific for the five cloned somatostatin receptor subtypes. Colocalization of these somatostatin receptors with the four major islet hormones--insulin, glucagon, somatostatin, and pancreatic polypeptide--was studied in normal human endocrine pancreatic cells, using double-immunofluorescence staining. High expression of somatostatin receptor subtypes 1, 3, and 4 was found in all endocrine pancreatic cells. Somatostatin receptor subtype 2 was frequently expressed in alpha and beta cells, whereas expression was low in pancreatic polypeptide cells and intermediate in delta cells. Somatostatin receptor subtype 5 was expressed in most beta and delta cells but almost absent in alpha and pancreatic polypeptide cells. There is a variability in the normal expression of somatostatin receptor subtypes among the different human endocrine pancreatic cells. Knowledge of this expression and the physiological function mediated by these receptors will be valuable in the future when considering treatment of endocrine disorders.

Cell mediated immunity and specific IgG1 and IgG2 antibody response in natural and experimental canine leishmaniosis.

In the present study, we have followed up Leishmania infantum infection in dogs: (1) naturally infected; (2) experimentally infected with amastigotes; and (3) experimentally infected with culture promastigotes. The main objective was to evaluate the differences of the humoral and cellular immune responses of each group. Sera from 12 beagle dogs were analysed for total anti-leishmanial antibodies and IgG1 and IgG2 subclasses by enzyme-linked immunosorbent assay (ELISA). Lymphoproliferation to L. infantum antigen was also performed. All naturally infected animals were symptomatic with a marked humoral response. Dogs inoculated with amastigotes were asymptomotic and presented lower antibody titres than naturally infected. Dogs inoculated with culture promastigotes were asymptomotic with no significant humoral response. Strong proliferative responses to Leishmania antigen was observed in dogs inoculated with promastigotes. In our experimental model, IgG1 antibody levels presented a similar pattern in all infected animals, and IgG2 reactivity was high in naturally infected dogs.

The enterochromaffin cells in the mouse gastrointestinal tract after streptozotocin treatment.

Effects of 5 weeks streptozotocin treatment on the mouse gastrointestinal tract were studied with special emphasis on enterochromaffin (EC) cells. In the streptozotocin-treated animals the frequency of EC cells was reduced in the antral area and in the small intestine, but increased in the colon. The length of the intestinal tract and the mucosal thickness were increased in these animals. Possible mechanisms underlying these abnormalities of the EC cell system are discussed.

Identification and characterization of enterochromaffin cells with different staining techniques.

The staining characteristics of the enterochromaffin cell system in the human and rat gastrointestinal tract were studied with 3 histochemical procedures--the Masson method, formaldehyde-induced fluorescence (FIF), and immunocytochemical techniques using both monoclonal and polyclonal serotonin antisera. A simple modification of the Masson technique is described and compared with several other modifications of the same technique. The specificities of the 2 antisera were examined with regard to serotonin and other monoamines and their albumin conjugates, monoamine precursors and metabolites, and also to serotonin-related substances. In the intestine, with very few exceptions the same cells reacted positively with the 3 staining methods, but often serotonin immunoreactivity was observed in larger cytoplasmic areas than the other 2 staining reactions. In the antral mucosa, a cell population with the same staining characteristics as mentioned above was seen, but 2 further cell populations were identified--one displaying FIF-argentaffin reactions but no serotonin immunoreactivity, and the other showing the opposite characteristics. At immunostaining, a discrepancy in the staining results between the 2 antisera was noted--a larger number of antral serotonin immunoreactive cells being demonstrated with the polyclonal than with the monoclonal antiserum; this discrepancy disappeared when the polyclonal antiserum was pretreated with C-terminal tetragastrin. A minor discrepancy in the number of stained cells was still observed between the use of immunostaining with monoclonal or with tetragastrin-pretreated polyclonal antiserum, and the use of the FIF-argentaffin reactions. The reason for this discrepancy is unclear.