Histologic Evaluation of the Osteoinductive Property of Autogenous Demineralized Dentin Matrix on Surgical Bone Defects in Rabbit Skulls Using Human Amniotic Membrane for Guided Bone Regeneration

The aim of this investigation was to evaluate the osteoinductive property of autogenous demineralized dentin matrix (ADDM) on experimental surgical bone defects in the parietal bone of rabbits using the guided bone regeneration (GBR) technique incorporating human amniotic membrane (HAM). Thirty-six rabbits were divided into 2 groups, HAM and ADDM+HAM. It was possible to conclude that HAM did not interfere with bone repair and was resorbed. Slices of ADDM induced direct bone formation and were incorporated by the newly formed bone tissue and remodeled. The bone defects healed faster in the ADDM+HAM group than in the group with HAM only.

Genitalia, acoustics and chromosomes of Urogryllus toledopizai (Orthoptera: Gryllidae)

Morphological structures of male and female genitalia and the pars stridens of a population of Urogryllus toledopizai Mello (1988) as well as the karyotype of the species are illustrated. The characteristics of song signals and associated behavior are described and information on the seasonality of the species is provided. Disagreements with the results published by other authors are discussed.

Cytogenetic analysis of five species of the subfamily Tetragonopterinae (Teleostei, Characiformes, Characidae)

The subfamily Tetragonopterinae is composed by a large number of species distributed in South and Central America. This subfamily has many taxonomic and phylogenetic problems, being considered by several authors as an artificial group. With the objective to better understanding the relationships among the components of this fish group, cytogenetic studies were conduced on five species of Tetragonopterinae. Astyanax janeiroensis had 2n=50 chromosomes (6M+14SM+14ST+16A), Hyphessobrycon reticulatus had 2n=50 chromosomes (14M+20SM+16ST), Hollandichthys multifasciatus had 2n=50 chromosomes (10M+12SM+28ST), Ctenobrycon hauxwellianus had 2n=50 chromosomes (10M+6SM+34ST), and Phenacogaster cf. pectinatus had 2n=46 chromosomes (12M+2ST+32A). Only A. janeiroensis had multiple NORs, while all other species had simple NORs. Small heterochromatic blocks were observed in the chromosomes of all species in a pericentromeric position. A. janeiroensis also had some chromosomes with large heterochromatic blocks in a terminal position and a pair with an interstitial block. The karyotypic evolution of each genus is discussed.

Treatment Effects Produced by Fränkel Appliance in Patients with Class II, Division 1 Malocclusion

The purpose of this investigation was to evaluate the dentoalveolar and skeletal cephalometric changes produced by the Fränkel appliance in individuals with a Class II, division 1 malocclusion. Lateral cephalograms of 44 patients of both sexes were divided in two groups of 22 each. The control group was comprised of untreated Class II children with an initial mean age of eight years and seven months who were followed without treatment for a period of 13 months. The Fränkel group had an initial mean age of nine years and was treated for a mean period of 17 months. Lateral cephalometric headfilms of each patient were obtained at the beginning and end of treatment. The Fränkel appliance produced no significant changes in maxillary growth during the evaluation period, while a statistically significant increase in mandibular length was observed. The maxillomandibular relationship improved mostly because of an increase in mandibular length. In addition, there were no statistically significant differences in the craniofacial growth direction between the Fränkel and the control group, both showing a slight downward rotation of the palatal plane. The Fränkel appliance produced a labial tipping of the lower incisors and a lingual inclination of the upper incisors as well as a significant increase in mandibular posterior dentoalveolar height. It was concluded that the main effects of the Fränkel appliance during this time period were mostly dentoalveolar with a smaller but significant skeletal mandibular effect.

EM evaluation of bacterial biofilm and microorganisms on the apical external root surface of human teeth

The aim of this study was to evaluate the presence of bacterial biofilm on the external surface of the root apex in teeth with pulp necrosis, with and without radiographically visible periapical lesions, and in teeth with a vital pulp. Twenty-one teeth were extracted, eight with pulp necrosis and periapical lesions, eight with pulp necrosis without radiographically visible periapical lesions, and five with a vital pulp. The roots were sectioned, and the root apexes (+/- 3 mm) were processed for scanning electron microscope evaluation. The surface of the apical root was evaluated for the presence of microorganisms, root resorption, and biofilm. There were no microorganisms on the apical root surface of either teeth with pulp vitality or with pulp necrosis with no radiographically visible periapical lesions. Microorganisms were always present in teeth with pulp necrosis and radiographically visible periapical lesions. These included cocci, bacilli, and filaments and the presence of an apical biofilm. Apical biofilm is clinically important because microbial biofilms are inherently resistant to antimicrobial agents and cannot be removed by biomechanical preparation alone. This may cause failure of endodontic treatment as a consequence of persistent infection.

Taxonomic importance of the prefemoral process of the first pair of legs in males of the genus Pseudonannolene (Diplopoda, Spirostreptida)

The first pair of legs in males of some groups of diplopods is modified, having a large coxa and oral projecting prefemoral process; in the family Spirostreptidae (Spirostreptida) this structure has relevance on the genus level. This paper discusses the shape, presence, and absence of the prefemoral process in the genus Pseudonannolene (Pseudonannolenidae, Spirostreptida) and suggests that this structure is an important taxonomic character in this genus and can be used for distinguishing some species.

Cloning, overexpression, and purification of functional human purine nucleoside phosphorylase

Purine nucleoside phosphorylase (PNP) catalyzes the phosphorolysis of the N-ribosidic bonds of purine nucleosides and deoxynucleosides. A genetic deficiency due to mutations in the gene encoding for human PNP causes T-cell deficiency as the major physiological defect. Inappropriate activation of T-cells has been implicated in several clinically relevant human conditions such as transplant tissue rejection, psoriasis, rheumatoid arthritis, lupus, and T-cell lymphomas. Human PNP is therefore a target for inhibitor development aiming at T-cell immune response modulation. In addition, bacterial PNP has been used as reactant in a fast and sensitive spectrophotometric method that allows both quantitation of inorganic phosphate (Pi) and continuous assay of reactions that generate P i such as those catalyzed by ATPases and GTPases. Human PNP may therefore be an important biotechnological tool for P i detection. However, low expression of human PNP in bacterial hosts, protein purification protocols involving many steps, and low protein yields represent technical obstacles to be overcome if human PNP is to be used in either high-throughput drug screening or as a reagent in an affordable P i detection method. Here, we describe PCR amplification of human PNP from a liver cDNA library, cloning, expression in Escherichia coli host, purification, and activity measurement of homogeneous enzyme. Human PNP represented approximately 42% of total soluble cell proteins with no induction being necessary to express the target protein. Enzyme activity measurements demonstrated a 707-fold increase in specific activity of cloned human PNP as compared to control. Purification of cloned human PNP was achieved by a two-step purification protocol, yielding 48 mg homogeneous enzyme from 1 L cell culture, with a specific activity value of 80 U mg -1. © 2002 Elsevier Science (USA). All rights reserved.

Collagen fibers in human prostatic lesions: histochemistry and anisotropies

The present study focuses on establishing patterns of collagen fibers distribution in prostatic nodular hyperplasia and adenocarcinomas, in comparison with the normal tissue. Sections of prostatic transurethral resection were subjected to Gömöri's method for collagen fibers and reticulin and analyzed under ordinary and polarized light microscopy. Controls and hyperplastic regions present collagen fibers with variable thickness that run in different directions, establishing a tridimensional network. These fibers exhibit birefringence and dichroism thus demonstrating their fibrillar integrity. On the other hand, increased variability in collagen fiber distribution and anisotropical properties occur in adenocarcinomas evaluated in accordance with the Gleason's score. In some of their areas, a well-defined collagen network delimitates the base of transformed epithelial cells whereas in other areas the collagen fibers are disorganized and do not establish a boundary between the epithelial structures and the stroma. In these areas, collagen is found in the stroma. It was also observed that adenocarcinoma tumor cells rest on a scaffold of thin and dendritic collagen fibers. Collagen fibers of the prostatic stroma of the adenocarcinomas may show a modification in arrangement and fibrillar compactness. In prostatic nodular hyperplasia, there is no change in collagen molecular integrity, since collagen affinity for silver and collagen birefringence are similar to controls. In adenocarcinoma with high dedifferentiation degree, thin and branched strongly argyrophilic and birefringent collagen fibers are detected in regions of cell proliferation. In the adjacent stroma, hyaline plaques are indicative of matrix degradation or remodellation.

Niche-specific association of Aeromonas ribotypes from human and environmental origin

A total of 88 Aeromonas isolates from distinct locations and sources (39 from extraintestinal infections, 31 from diarrhoeic, ten from non-diarrhoeic faeces, all human, and eight from fresh water) were subjected to phenospecies identification, serotyping, ribotyping and detection of some virulence markers. The strains belonged to four different phenospecies marked by 19 O serogroups and 38 ribotypes. No strong correlation between these parameters was found, and no group, as defined by the typing methods, could be characterized with a particular set of virulence markers. There was a clear association of ribotypes with the source of the strains. Cluster analysis allowed the identification of a complex of ribotypes belonging to distinct but related sources, including clinical and environmental isolates. These results suggest that ribotyping may be an epidemiological tool suitable for the study of Aeromonas infections.

Computer-assisted analysis of p53 and PCNA expression in oral lesions infected with human papillomavirus

OBJECTIVE: To carry out a retrospective study to determine whether human papillomavirus (HPV) infection and immunohistochemical expression of p53 and proliferating cell nuclear antigen (PCNA) are related to the risk of oral cancer. STUDY DESIGN: Fifty-seven oral biopsies, consisting of 30 oral squamous papillomas (OSPs) and 27 oral squamous cell carcinomas (OSCCs) were tested for the presence of HPV 6/11 and 16/18 by in situ hybridization using catalyzed signal amplification and in situ hybridization. p53 And PCNA expression was analyzed by immunohistochemistry and evaluated quantitatively by image analysis. RESULTS: Nineteen of the 57 oral lesions (33.3%) were positive for HPV. HPV 6/11 was found in 6 of 30 (20%) OSPs and 1 of 27 (3.7%) OSCCs. HPV 16/18 was found in 10 of 27 (37%) OSCCs and 2 of 30 (6.7%) OSPs. Sixteen of the 19 HPV-positive cases (84.2%) were p53 negative; 5 (9%) were HPV 6/11 and 11 (19%) HPV 16/18, with an inverse correlation between the presence of HPV DNA and p53 expression (P=.017, P < .05). PCNA expression appeared in 18 (94.7%) of HPV positive cases, showing that HPV 16/18 was associated with intensity of PCNA expression and with OSCCs (P=.037, P < .05). CONCLUSION: Quantitative evaluation of p53 by image analysis showed an inverse correlation between p53 expression and HPV presence, suggesting protein degradation. Image analysis also demonstrated that PCNA expression was more intense in HPV DNA 16/18 OSCCs. These findings suggest involvement of high-risk HPV types in oral carcinogenesis.

First cytogenetic description of the species Rhamdella microcephala (Pisces, Hepapteridae)

Cytogenetic studies were carried out on Rhamdella microcephala collected from the headwaters of the Machado river, a tributary of the Sapucaí river, Minas Gerais, Brazil. Sixteen specimens exhibited 2n=56 chromosomes, with 18 metacentrics, 30 submetacentrics and 8 subtelocentric-acrocentrics (FN=84). The nucleolar organizing regions (NORs) were identified in the interstitial position on the long arm of a submetacentric pair (pair 12). Chromomycin A3 staining evidenced only the NOR-bearing segments. Positive C-band segments were identified in a pericentromeric position in most of the chromosomes and the NOR-bearing segments were also C-band positive. Some aspects related to the chromosomal characteristics of Rhamdella microcephala are discussed.

Molecular characterization and chromosomal localization of two families of satellite DNA in Prochilodus lineatus (Pisces, Prochilodontidae), a species with B chromosomes

Prochilodus lineatus, an abundant species in the Mogi-Guaçu river basin, represents a large part of the region's fishing potential. Karyotypic analyses based on classic cytogenetic techniques have revealed the presence of 54 metasubmetacentric type chromosomes, together with the occurrence of small supernumerary chromosomes with intra and interindividual variations. This paper describes the genomic organization of two families of satellite DNA in the P. lineatus genome. The chromosomal localization these two repetitive DNA families through fluorescence in situ hybridization (FISH) demonstrated that the SATH1 satellite DNA family, composed of approximately 900 bp, was located in the pericentromeric region of a group of chromosomes of the standard complement, as well as on all the B chromosomes. The SATH2 satellite family has a monomeric unit of 441 bp and was located in the pericentromeric regions of some chromosomes of the standard complement, but was absent in the B chromosomes. Double FISH analyses showed that these two families participate jointly in the pericentromeric organization of several chromosomes of this species. The data obtained in this study support the hypothesis that the B chromosomes derive from chromosomes of the standard complement, which are carriers of the SATH1 satellite DNA.

Nuclear phenotypes and morphometry of human secretory prostate cells: A comparative study of benign and malignant lesions in Brazilian patients

Prostatic lesions in Brazilian patients with benign prostatic hyperplasia (BPH, 26 cases) or adenocarcinoma (AC, 25 cases) were compared by qualitative microscopy and morphometric analysis. In 12 cases of BPH, prostate regions with no histological alterations were considered as controls (Ct). Archival material consisted of formalin-fixed, paraffin-embedded specimens obtained from prostatic transurethral resection and radical prostatectomy. Haematoxylin/eosin (HE)-stained sections were used to estimate the nuclear areas, perimeters and form factor values. HE-stained sections from AC specimens were also used for Gleason grading. BPH, AC and Ct could be discriminated by their nuclear areas and nuclear perimeters, but not by the nuclear form factor parameter. No significant differences were found when the AC data were compared using the combined version or the predominant grade version of the Gleason score (p = 0.8380 for nuclear area; p = 0.6076 for nuclear perimeter; p = 0.9202 for nuclear form factor; n = 200 nuclei per patient). This finding indicates that there is extensive heterogeneity in the size and shape of the nucleus in AC cells. These results also show that although the nuclear morphometry served to discriminate BPH and AC from each other and from Ct, it was not sufficient to correlate AC lesions with their respective Gleason scores in the human population analyzed.

The natural history of carpal tunnel syndrome: A study of 20 hands evaluated 4 to 9 years after initial diagnosis

Carpal tunnel syndrome (CTS) is the most frequent entrapment neuropathy. In the last decade several papers have been published on epidemiology, clinical aspects, diagnosis, and treatment, but little is known about its natural history. The objective of this work was to study the natural history of CTS syndrome. From 358 patients with clinical and conduction study diagnosis of CTS, 12 cases were identified that had refused surgical treatment, had not used anti-inflammatory medications, and had not undergone orthopaedic procedures, such as immobilization or anaesthetic infiltration. These 12 patients have 20 compromised hands which have been followed up for between 4 and 9 years. In all cases sensory and motor conduction studies were performed on the median nerve, at the beginning and end of follow-up period. Electrical improvement was marked in 5 hands and slight in 3; there was no significant change in 10, and deterioration in 2. As 8 hands (7 patients) showed improved clinical symptoms and conduction studies over several years, this brings the universally accepted procedure of surgical treatment into doubt.

Genetic relatedness of Plasmodium falciparum isolates and the origin of allelic diversity at the merozoite surface protein-1 (MSP-1) locus in Brazil and Vietnam

Background: Despite the extensive polymorphism at the merozoite surface protein-1 (MSP-1) locus of Plasmodium falciparum, that encodes a major repetitive malaria vaccine candidate antigen, identical and nearly identical alleles frequently occur in sympatric parasites. Here we used microsatellite haplotyping to estimate the genetic distance between isolates carrying identical and nearly identical MSP-1 alleles. Methods: We analyzed 28 isolates from hypoendemic areas in north-western Brazil, collected between 1985 and 1998, and 23 isolates obtained in mesoendemic southern Vietnam in 1996. MSP-1 alleles were characterized by combining PCR typing with allele-specific primers and partial DNA sequencing. The following single-copy microsatellite markers were typed: Polyα, TA42 (only for Brazilian samples), TA81, TA1, TA87, TA109 (only for Brazilian samples), 2490, ARAII, PfG377, PfPK2, and TA60. Results: The low pair-wise average genetic distance between microsatellite haplotypes of isolates sharing identical MSP-1 alleles indicates that epidemic propagation of discrete parasite clones originated most identical MSP-1 alleles in parasite populations from Brazil and Vietnam. At least one epidemic clone propagating in Brazil remained relatively unchanged over more than one decade. Moreover, we found no evidence that rearrangements of MSP-1 repeats, putatively created by mitotic recombination events, generated new alleles within clonal lineages of parasites in either country. Conclusion: Identical MSP-1 alleles originated from co-ancestry in both populations, whereas nearly identical MSP-1 alleles have probably appeared independently in unrelated parasite lineages.