Low Expression of Human Histocompatibility Soluble Leukocyte Antigen-G (HLA-G5) in Invasive Cervical Cancer With and Without Metastasis, Associated With Papilloma Virus (HPV)

Human leukocyte antigen-G (HLA-G) is a non-classical major histocompatibility complex class lb molecule that acts as a specific immunosuppressor. Some studies have demonstrated that human papillomavirus (HPV) seems to be involved in lower or absent HLA-G expression, particularly in cervical cancer. In this study, we performed a cross-sectional study, systematically comparing the qualitative expression of the HLA-G5 isoform in invasive cervical carcinoma (ICC), stratifying patients according to the presence [ICC with metastasis (ICC(W))) and absence [ICC without metastasis (ICC(WT))] of metastasis, correlating these findings with interference of HPV and demographic and clinical variables. Seventy-nine patients with a diagnosis of ICC were stratified into two groups: ICC(WT) (n=52 patients) and ICC(W) (n=27). Two biopsies were collected from each patient (one from the tumor lesion and one from a lymph node). Immunohistochemistry analyses were performed for the HLA-G5 isoform, for HPV detection, and virus typing. HLA-G5 isoform molecules were detected in 25 cases (31.6%), 17 (32.7%) without metastasis and 8 (29.6%) with metastasis. HPV was detected in the cervical lesions of 74 patients (93.7%), but low expression of the HLA-G5 isoform was observed in all HPV-related cases. These findings are important; however, additional studies are necessary to identify the influence of HPV with HLA-G5 isoform expression on invasive cervical malignancies. (J Histochem Cytochem 58:405-411, 2010)

Pompe disease in a Brazilian series: clinical and molecular analyses with identification of nine new mutations

Pompe disease (glycogen storage disease type II or acid maltase deficiency) is an inherited autosomal recessive deficiency of acid alpha-glucosidase (GAA), with predominant manifestations of skeletal muscle weakness. A broad range of studies have been published focusing on Pompe patients from different countries, but none from Brazil. We investigated 41 patients with either infantile-onset (21 cases) or late-onset (20 cases) disease by muscle pathology, enzyme activity and GAA gene mutation screening. Molecular analyses identified 71 mutant alleles from the probands, nine of which are novel (five missense mutations c.136T > G, c.650C > T, c.1456G > C, c.1834C > T, and c.1905C > A, a splice-site mutation c.1195-2A > G, two deletions c.18_25del and c.2185delC, and one nonsense mutation c.643G > T). Interestingly, the c.1905C > A variant was detected in four unrelated patients and may represent a common Brazilian Pompe mutation. The c.2560C > T severe mutation was frequent in our population suggesting a high prevalence in Brazil. Also, eight out of the 21 infantile-onset patients have two truncating mutations predicted to abrogate protein expression. Of the ten late-onset patients who do not carry the common late-onset intronic mutation c.-32-13T > G, five (from three separate families) carry the recently described intronic mutation, c.-32-3C > A, and one sibpair carries the novel missense mutation c.1781G > C in combination with known severe mutation c.1941C > G. The association of these variants (c.1781G > C and c.-32-3C > A) with late-onset disease suggests that they allow for some residual activity in these patients. Our findings help to characterize Pompe disease in Brazil and support the need for additional studies to define the wide clinical and pathological spectrum observed in this disease.

Sensitivity and uncertainty analyses for burden of disease and risk factor estimates

Epidemiological studies report confidence or uncertainty intervals around their estimates. Estimates of the burden of diseases and risk factors are subject to a broader range of uncertainty because of the combination of multiple data sources and value choices. Sensitivity analysis can be used to examine the effects of social values that have been incorporated into the design of the disability–adjusted life year (DALY). Age weight, where a year of healthy life lived at one age is valued differently from at another age, is the most controversial value built into the DALY. The discount rate, which addresses the difference in value of current versus future health benefits, also has been criticized. The distribution of the global disease burden and rankings of various conditions are largely insensitive to alternate assumptions about the discount rate and age weighting. The major effects of discounting and age weighting are to enhance the importance of neuropsychiatric conditions and sexually transmitted infections. The Global Burden of Disease study also has been criticized for estimating mortality and disease burden for regions using incomplete and uncertain data. Including uncertain results, with uncertainty quantified to the extent possible, is preferable, however, to leaving blank cells in tables intended to provide policy makers with an overall assessment of burden of disease. No estimate is generally interpreted as no problem. Greater investment in getting the descriptive epidemiology of diseases and injuries correct in poor countries will do vastly more to reduce uncertainty in disease burden assessments than a philosophical debate about the appropriateness of social value

Functional and bioinformatics analyses reveal conservation of Cis-regulatory elements between sciaridae and Drosophilidae

The sciarid DNA puff C4 BhC4-1 gene is amplified and transcribed in salivary glands at the end of the larval stage. In transgenic Drosophila, the BhC4-1 promoter drives transcription in prepupal salivary glands and in the ring gland of late embryos. A bioinformatics analysis has identified 162 sequences similar to distinct regions of the BhC4-1 proximal promoter, which are predominantly located either in 5` or 3` regions or introns in the Drosophila melanogaster genome. A significant number of the identified sequences are found in the regulatory regions of Drosophila genes that are expressed in the salivary gland. Functional assays in Drosophila reveal that the BhC4-1 proximal promoter contains both a 129 bp (-186/-58) salivary gland enhancer and a 67 bp (-253/-187) ring gland enhancer that drive tissue specific patterns of developmentally regulated gene expression, irrespective of their orientation.

Canine Distemper Virus in a Crab-eating Fox (Cerdocyon thous) in Brazil: Case Report and Phylogenetic Analyses

Although canine distemper is enzootic worldwide and has a wide host range, there are no reports of canine distemper virus there are no reports of canine distemper virus provide information on virus phylogeny and histopathologic lesions. The objective of this study is report and describe canine distemper in a crab-eating fox (C. thous), with a focus oil the phylogeny of the virus strain and the histopathologic lesions in the animal.

Adhesives bonded to erbium:yttrium-aluminum-garnet laser-irradiated dentin: transmission electron microscopy, scanning electron microscopy and tensile bond strength analyses

The aim of this in vitro study was to investigate the effect of erbium:yttrium-aluminum-garnet (Er:YAG) laser irradiation on dentinal collagen by transmission electron microscopy and to analyze the resin-dentin interface by scanning electron microscopy. A tensile bond strength test was also applied. Specimens from 69 sound human third molars were randomly divided into three groups: control (no laser), and two irradiated groups, laser 250 (250 mJ/2 Hz) and laser 400 (400 mJ/4 Hz). Then, specimens were restored with two adhesive systems, an etch-and-rinse or a self-etch system. Although ultrastructural examination showed a modified surface in the irradiated dentin, there was no statistical difference in bond strength values between the laser groups and controls (P < 0.05). In conclusion, the use of Er:YAG laser for ablating human dentin did not alter the main adhesion parameters when compared with those obtained by conventional methods, thus reinforcing its use in restorative dentistry.

A critical view on biaxial and short-beam uniaxial flexural strength tests applied to resin composites using Weibull, fractographic and finite element analyses

Objective. To evaluate the biaxial and short-beam uniaxial strength tests applied to resin composites based upon their Weibull parameters, fractographic features and stress distribution. Methods. Disk- (15 mm x 1 mm) and beam-shaped specimens (10 mm x 2 mm x 1 mm) of three commercial composites (Concept/Vigodent, CA; Heliomolar/Ivoclar-Vivadent, HE; Z250/3M ESPE, FZ) were prepared. After 48h dry storage at 37 degrees C, disks and beams were submitted to piston-on-three-balls (BI) and three-point bending (UNI) tests, respectively. Data were analyzed by Weibull statistics. Fractured surfaces were observed under stereomicroscope and scanning electron microscope. Maximum principal stress (sigma(1)) distribution was determined by finite element analysis (FEA). Maximum sigma(1-BI) and sigma(1-UNI) were compared to FZ strengths calculated by applying the average failure loads to the analytical equations (sigma(a-BI) and sigma(a-UNI)). Results. For BI, characteristic strengths were: 169.9a (FZ), 122.4b (CA) and 104.8c (HE), and for UNI were: 160.3a (FZ), 98.2b (CA) and 91.6b (HE). Weibull moduli ( m) were similar within the same test. CA and HE presented statistically higher m for BI. Surface pores ( BI) and edge flaws ( UNI) were the most frequent fracture origins. sigma(1-BI) was 14% lower than sigma(a-BI.) sigma(1-UNI) was 43% higher than sigma(a-UNI). Significance. Compared to the short-beam uniaxial test, the biaxial test detected more differences among composites and displayed less data scattering for two of the tested materials. Also, biaxial strength was closer to the material`s strength estimated by FEA. (C) 2009 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Bacterial leakage in obturated root canals-part 2: a comparative histologic and microbiologic analyses

Objective. In this study, presence of dentin infection in root canals, obturated with 4 techniques submitted to the bacterial leakage test, was evaluated using histologic methods. Study design. The canals of palatal roots of 160 molars were instrumented and divided into different groups, according to the obturation technique used (lateral condensation, MicroSeal system, Touch `n Heat + Ultrafil, and Tagger`s hybrid technique) and extent of the remaining obturation material (5 mm and 10 mm). Ten additional roots were used as control samples. The roots were sterilized in ethylene oxide and mounted on a device for evaluation of bacterial leakage using the bacteria Enterococcus faecalis for 120 days. After the leakage test, roots were microscopically analyzed for the presence of dentin infection in the root canals and dentinal tubules. Results. A total of 154 specimens were analyzed using both methodologies in the experimental groups; 50 root canals (32.4%) showed bacterial leakage at the end of the experimental period, and 118 (76.6%) showed the presence of bacteria in the root canals using the histologic criteria. The lateral condensation technique allowed lower penetration of bacteria in the root canals and dentinal tubules, followed by Touch `n Heat + Ultrafil, MicroSeal, and Tagger`s hybrid technique, which allowed significantly greater penetration of bacteria. Root canals with 10 mm of remaining obturation material presented similar bacterial penetration as root canals with 5 mm. Conclusions. Even when an adequate seal of the apical foramen was shown by the absence of turbidity in the bacterial leakage test, E. faecalis dentin infection was present in a high percentage of the root canals after 120 days of root filling exposure to the bacteria. Tagger`s hybrid technique presented greater quantity of bacteria in histologic sections than root canals obturated with the other techniques. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: 788-794)

Histological and molecular temporomandibular joint analyses after mandibular advancement surgery: study in minipigs

Objectives. The objective of this study was to elucidate the changes occurring in the temporomandibular joint (TMJ) after surgical mandibular advancement with different fixation techniques: bicortical screws (rigid fixation) and miniplates (semi-rigid fixation). Study design. Eighteen minipigs were equally and randomly divided into 3 groups: Group I (control), nonoperated animals; Group II, animals submitted to surgical advancement surgery and osteosynthesis by bicortical screws; and Group III, animals submitted to surgical advancement surgery and osteosynthesis by miniplates. Four months after the surgeries, the presence of interleukin (IL)-6 and IL-10 in synovial fluid samples was assessed in ELISA experiments. TMJs were histologically prepared. Results. Higher levels of IL-10 (P = .0436) were found for Group II. Descriptive histological analysis was compatible with the ELISA findings. Conclusions. Rigid fixation evokes more pronounced signs of bone remodeling in the TMJ, whereas malleable fixation promotes a more intense inflammatory activity. Therefore, rigid fixation seems to transmit a higher impact of postoperative masticatory forces to the TMJ.

Morphological, Morphometrical and Histochemical Study of the Lining and Glandular Epithelia of the Tongue of the Bullfrog Rana catesbeiana

The dorsal surface of the tongue of the bullfrog, Rana catesbeiana, has simple columnar epithelium with a few ciliated cells and goblet cells. The entire surface is covered with numerous filiform papillae and few fungiform. Filiform papillae have a simple columnar epithelium with secretory cells, while the fungiform have a sensory disc on their upper surface the lined by a stratified columnar epithelium with basal, peripheral, glandular and receptor cells. Over the dorsal lingual surface there are numerous winding tubular glands, which penetrate deeply into the muscle of the tongue, mingling with the fibers. The gland epithelium is cylindrical with secretory and supporting cells. The first are absolute on the basis of the gland and the latter are rare in the upper third. The ventral surface of the tongue is lined by a stratified epithelium, with the presence of goblet cells, with ciliated cells among them. Morphometrically, lingual glands varies in length, according to their location: shorter in the anterior region of the tongue (330 mu m) than in the posterior region (450 mu m). Secretory cells of the anterior lingual glands are smaller (1457.7 mm(3)) than the posterior ones (2645.9 mu m(3)). The same can be said of the cell nuclei, 130.0 mu m(3) for the anterior glands and 202.3 mu m(3) for the posterior ones. Secretory cells of the lingual glands contain substances rich in protein and neutral mucopolysaccharides, which characterize the seromucous type. Goblet cells of the dorsal and ventral surface epithelia secrete neutral mucopolysaccharides and proteins, and can be characterized as type G1 cells, and the supporting cells of the superficial glands of the fungiform papillae secrete a mucus rich in neutral mucopolysaccharides, sulfomucins and sialomucins.

Composite filling removal with erbium:yttrium-aluminum-garnet laser: morphological analyses

Considering the increase in esthetic restorative materials and need for improvement in unsatisfactory restoration substitution with minimal inadvertent removal of healthy tissues, this study assessed the efficacy of erbium:yttrium-aluminum-garnet (Er:YAG) laser for composite resin removal and the influence of pulse repetition rate on the morphological analyses of the cavity by scanning electron microscope. Composite resin fillings were placed in cavities (1.0 mm deep) prepared in bovine teeth, and the 75 specimens were randomly assigned to five groups according to the technique used for composite filling removal (high-speed diamond bur, group I, as a control, and Er:YAG laser, 250 mJ output energy and 80 J/cm(2) energy density, using different pulse repetition rates: group II, 2 Hz; group III, 4 Hz; group IV, 6 Hz; group V, 10 Hz). After the removal, the specimens were split in the middle, and we analyzed the surrounding and deep walls to check for the presence of restorative material. The estimation was qualitative. The surfaces were examined with a scanning electron microscope. The results revealed that the experimental groups presented bigger amounts of remaining restorative material. The scanning electron microscopy (SEM) analyses showed irregularities of the resultant cavities of the experimental groups that increased proportionally with increase in repetition rate.

Phenotypic and genetic analyses of a short measure of psychosis-proneness in a large-scale Australian twin study

Previous genetic analyses of psychosis proneness have been limited by their small sample size. For the purposes of large-scale screening, a 12-item questionnaire was developed through a two-stage process of reduction from the full Chapman and Chapman scales. 3685 individuals (including 1438 complete twin pairs) aged 18–25 years and enrolled in the volunteer Australian Twin Registry returned a mail questionnaire which included this psychosis proneness scale and the Eysenck Personality Questionnaire. Despite the brevity of the questionnaire, item and factor analysis identified four unambiguous and essentially uncorrelated scales. There were (1) Perceptual Aberration – Magical Ideation; (2) Hypomania – Impulsivity/Nonconformity; (3) Social Anhedonia and (4) Physical Anhedonia. Model-fitting analyses showed additive genetic and specific environmental factors were sufficient for three of the four scales, with the Social Anhedonia scale requiring also a parameter for genetic dominance. There was no evidence for the previously hypothesised sex differences in the genetic determination of psychosis-proneness. The potential value of multivariate genetic analysis to examine the relationship between these four scales and dimensions of personality is discussed. The growing body of longitudinal evidence on psychosis-proneness suggests the value of incorporating this brief measure into developmental twin studies.

Phylogenetic relationships of Trypanosoma chelodina and Trypanosoma binneyi from Australian tortoises and platypuses inferred from small subunit rRNA analyses

Trypanosome infections are often difficult to detect by conventional microscopy and their pleomorphy often confounds differential diagnosis. Molecular techniques are now being used to diagnose infections and to determine phylogenetic relationships between species. Complete small subunit rRNA gene sequences were determined for isolates of Trypanosoma chelodina from the Brisbane River tortoise (Emydura signata), the saw-shelled tortoise (Elseya latisternum), and the eastern snake-necked tortoise (Chelodina longicollis) from southeast Queensland, Australia. Partial sequence data were also obtained for T. binneyi from a platypus (Ornithorhynchus anatinus) from Tasmania. Phylogenetic relationships between T. chelodina, T. binneyi and other species were examined by maximum parsimony and likelihood methods. The Australian tortoise and platypus trypanosomes did not exhibit any close phylogenetic relationships with those of mammals, reptiles or amphibians, but were closely related to each other, and to fish trypanosomes. This contra-indicates their co-evolution with their vertebrate hosts but does not exclude co-evolution with different groups of invertebrate vectors, notably insects and leeches.

Assessing ecological impacts of shrimp and sewage effluent: Biological indicators with standard water quality analyses

Despite evidence linking shrimp farming to several cases of environmental degradation, there remains a lack of ecologically meaningful information about the impacts of effluent on receiving waters. The aim of this study was to determine the biological impact of shrimp farm effluent, and to compare and distinguish its impacts from treated sewage effluent. Analyses included standard water quality/sediment parameters, as well as biological indicators including tissue nitrogen (N) content, stable isotope ratio of nitrogen (delta N-15) and amino acid composition of inhabitant seagrasses, mangroves and macroalgae. The study area consisted of two tidal creeks, one receiving effluent from a sewage treatment plant and the other from an intensive shrimp farm. The creeks discharged into the western side of Moreton Bay, a sub-tropical coastal embayment on the east coast of Australia. Characterization of water quality revealed significant differences between the creeks, and with unimpacted eastern Moreton Bay. The sewage creek had higher concentrations of dissolved nutrients (predominantly NO3-/NO2- and PO43-, compared to NH4+ in the shrimp creek). In contrast, the shrimp creek was more turbid and had higher phytoplankton productivity. Beyond 750 m from the creek mouths, water quality parameters were indistinguishable from eastern Moreton Bay values. Biological indicators detected significant impacts up to 4 km beyond the creek mouths (reference site). Elevated plant delta N-15 values ranged from 10.4-19.6 parts per thousand at the site of sewage discharge to 2.9-4.5 parts per thousand at the reference site. The free amino acid concentration and composition of seagrass and macroalgae was used to distinguish between the uptake of sewage and shrimp derived N. Proline (seagrass) and serine (macroalgae) were high in sewage impacted plants and glutamine (seagrass) and alanine (macroalgae) were high in plants impacted by shrimp effluent. The delta N-15 isotopic signatures and free amino acid composition of inhabitant flora indicated that sewage N extended further from the creek mouths than shrimp N. The combination of physical/chemical and biological indicators used in this study was effective in distinguishing the composition and subsequent impacts of aquaculture and sewage effluent on the receiving waters. (C) 2001 Academic Press.

Amino acid residues in conodont elements

Thermally unaltered conodont elements, brachiopods. and vertebrates were analyzed with reverse phase high profile liquid chromatography to locate and quantify amino acid remnants of the original organic matrix in the fossils. No consistent similarities in amino acid content were found in conodont taxa. and criteria based on organic residues appear to have no taxonomic significance in the fossils tested from these localities. However, hydroxyproline. an amino acid that is found in the collagen molecules of animals. as well as in the glycoproteins in the cell walls and reproductive tissues of certain plants, is represented in most taxa. The organic matter retained in the impermeable crowns of conodont elements might have been derived originally from a form of collagen. Biochemical analyses. correlated with histochemical tests, demonstrate that organic matter is an integral part of the hyaline tissue of the element crown and not the result of surface contamination. Tests of a range of vertebrate and invertebrate fossil hard tissues produced similar results. The analyses indicate that hyaline tissue in the conodont element crown is not a form of vertebrate enamel. which contains no collagen. Albid tissue. with little or no organic content. is not a form of vertebrate bone or dentine, both based on collagen and low in mineral. Although these results do not help to determine the phylogenetic affinities of conodont animals, they indicate teat conodont elements do not contain hard tissues characteristic of vertebrate animals.