Effect of refrigeration systems upon frozen bull sperm viability assessed by computer-assisted sperm analysis and fluorescent probes

Sperm cryopreservation success depends upon the maintenance of spermatozoa fertility potential. Sperm cells must preserve both integrity and functionality of several cell structures. The stabilization phase must allow the exit of water from the sperm cells via osmosis. This study aimed to compare the effect of refrigeration in the commercial refrigerator (CR) and the transport/refrigeration box (TRB) upon the viability of frozen bull sperm diluted in three different extenders (A, B and C). Ten Nellore bulls, Bos taurus indicus maintained in Artificial Insemination Center were used and the spermatozoa samples was assessed for Plasma Membrane Integrity and CASA evaluation. The stabilization phase (5 degrees C/4 hours) was performed in the CR as well as in the TRB, and then samples were exposed to nitrogen vapor during 20 minutes and then plunged into nitrogen. The statistical analysis was done using the variance analysis and the significance level was set at 5%. In the CR the post-thawing parameters for PM and ALH were higher (p < 0.05) in the extender A (glicine egg-yolk) and extender B (glicine egg-free) when compared with extender C (TRIS egg-yolk). As for BCF, STR and LIN, the parameters were higher (p < 0.05) in extender B than in C. Samples that were stabilized in the TRB presented higher post-thawing parameters (p < 0.05) for PM and LIN in extender A and extender B when compared with C. BCF and STR parameters were higher (p < 0.05) in extemder B when compared with C. Extender B samples had higher (p < 0.05) PMI when stabilized in CR. The findings in this experiment enable us to say that both CR and TRB were effective in keeping the viability of post-thawing bull semen.

Computer-assisted analysis of cell proliferation markers in oral lesions

Abnormalities in any component of the cell cycle regulatory machine may result in oral. cancer, and markers of cell proliferation have been used to determine the prognosis of tumor progression. The aim of this study was to determine whether silver-stained nucleolar organizer region (AgNOR) and Ki-67 measurements could improve the assessment of growth rates in oral lesions. Eighty-three oral biopsies were studied, 20 of which were classified as fibrous inflammatory hyperplasia (FIH), 40 as leukoplakia (LKP) and 23 as oral. squamous cell carcinoma (OSCC). Within the LKP group, 22 out of 29 biopsies were diagnosed as non-dysplastic leukoplakia (LK) and 18 as dysplastic teukoptakia (DLK), presenting discrete, moderate and severe dysplasia. Ki-67 immunotabeting of the lesions increased steadily in the following order: FIH, DLK, LK and OSCC, indicating that Ki-67 is a good marker for predicting the protiferative fraction among benign, premalignant and malignant oral lesions. The median values of AgNOR parameters indicate that the morphometric index gives better results regarding the proliferative rate than the numerical one. A series of linear regressions between AgNOR parameters and Ki-67 showed positive associations. We conclude that a combination of Ki-67 and morphometric AgNOR analyses could be used as an aid in the determination of the protiferative status of oral epithelial. cells in oral cancer. (C) 2007 Elsevier GmbH. All rights reserved.

Galectin-3 Overexpression in Invasive Laryngeal Carcinoma, Assessed by Computer-assisted Analysis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A computer-assisted approach for chemotaxonomic studies - Diterpenes in Lamiaceae

This paper describes a new computer approach for chemotaxonomic studies. The methodology employed enables the search for chemical substructures as taxonomic descriptors using all expert system built with plant natural products. The operation of the system was tested with diterpenes as taxonomic markers in Lamiaceae. © 2001 Elsevier Science Ltd.

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.

Periodontal effects of rapid maxillary expansion with tooth-tissue-borne and tooth-borne expanders: A computed tomography evaluation

Introduction: The force delivered during rapid maxillary expansion (RME) produces areas of compression on the periodontal ligament of the supporting teeth. The resulting alveolar bone resorption can lead to unwanted tooth movement in the same direction. The purpose of this study was to evaluate periodontal changes by means of computed tomography after RME with tooth-tissue-borne and tooth-borne expanders. Methods: The sample comprised 8 girls, 11 to 14 years old, with Class I or II malocclusions with unilateral or bilateral posterior crossbites Four girls were treated with tooth-tissue-borne Haas-type expanders, and 4 were treated with tooth-borne Hyrax expanders. The appliances were activated up to the full 7-mm capacity of the expansion screw. Spiral CT scans were taken before expansion and after the 3-month retention period when the expander was removed. One-millimeter thick axial sections were exposed parallel to the palatal plane, comprising the dentoalveolar area and the base of the maxilla up to the inferior third of the nasal cavity. Multiplanar reconstruction was used to measure buccal and lingual bone plate thickness and buccal alveolar bone crest level by means of the computerized method. Results and Conclusions: RME reduced the buccal bone plate thickness of supporting teeth 0.6 to 0.9 mm and increased the lingual bone plate thickness 0.8 to 1.3 mm. The increase in lingual bone plate thickness of the maxillary posterior teeth was greater in the tooth-borne expansion group than in the tooth-tissue-borne group. RME induced bone dehiscences on the anchorage teeth's buccal aspect (7.1 ± 4.6 mm at the first premolars and 3.8 ± 4.4 mm at the mesiobuccal area of the first molars), especially in subjects with thinner buccal bone plates. The tooth-borne expander produced greater reduction of first premolar buccal alveolar bone crest level than did the tooth-tissue-borne expander. © 2006 American Association of Orthodontists.

Quantitative cell-cycle protein expression in oral cancer assessed by computer-assisted system

The knowledge of cell-cycle control has shown that the capacity of malignant growth is acquired by the stepwise accumulation of defects in specific genes regulating cell growth. Histologic diagnosis might be improved by a quantitative evaluation of more specific diagnosis biomarkers, which could help to precisely identify pre-malignant and malignant oral lesions. The aim of the present study is to evaluate whether computer-based quantitative assessment of p53, PCNA and Ki-67 immunohistochemical expression, could be used clinically to foresee the risk of oral malignant transformation. This retrospective study was carried out in ninety-five oral biopsies, 27 were classified as fibrous inflammatory hyperplasia, 40 as leukoplakia and 28 as oral squamous cell carcinoma. Sixteen out of the 40 leukoplakia were diagnosed as non-dysplastic leukoplakia, the other 24 being dysplastic leukoplakia, of which 50.0% were classified as moderate to severe dysplasia. Comparison of the four groups of oral tissues showed significant rises in p53 and Ki-67 positivity index, which increased steadily in the order benign, pre-malignant, and malignant. In contrast, it was not possible to relate higher PCNA levels with pre-malignant and malignant oral lesions. We therefore conclude that PCNA immunohistochemistry expression is probably an inappropriate marker to identify oral carcinogenesis, whereas joint quantitative evaluation of p53 and Ki-67, appears to be useful as a tumor marker, providing a pre-diagnostic estimate of the potential for cell-cycle deregulation of the oral proliferate status.

Comparison of different designs of implant-retained overdentures and fixed full-arch implant-supported prosthesis on stress distribution in edentulous mandible - A computed tomography-based three-dimensional finite element analysis

A finite element analysis was used to compare the effect of different designs of implant-retained overdentures and fixed full-arch implant-supported prosthesis on stress distribution in edentulous mandible. Four models of an human mandible were constructed. In the OR (O'ring) group, the mandible was restored with an overdenture retained by four unsplinted implants with O'ring attachment; in the BC (bar-clip) -C and BC groups, the mandibles were restored with overdentures retained by four splinted implants with bar-clip anchor associated or not with two distally placed cantilevers, respectively; in the FD (fixed denture) group, the mandible was restored with a fixed full-arch four-implant-supported prosthesis. Models were supported by the masticatory muscles and temporomandibular joints. A 100-N oblique load was applied on the left first molar. Von Mises (σvM), maximum (σmax) and minimum (σmin) principal stresses (in MPa) analyses were obtained. BC-C group exhibited the highest stress values (σvM=398.8, σmax=580.5 and σmin=-455.2) while FD group showed the lowest one (σvM=128.9, σmax=185.9 and σmin=-172.1). Within overdenture groups, the use of unsplinted implants reduced the stress level in the implant/prosthetic components (59.4% for σvM, 66.2% for σmax and 57.7% for σmin versus BC-C group) and supporting tissues (maximum stress reduction of 72% and 79.5% for σmax, and 15.7% and 85.7% for σmin on the cortical and trabecular bones, respectively). Cortical bone exhibited greater stress concentration than the trabecular bone for all groups. The use of fixed implant dentures and removable dentures retained by unsplinted implants to rehabilitate edentulous mandible reduced the stresses in the periimplant bone tissue, mucosa and implant/prosthetic components. © 2013 Elsevier Ltd.

A Comparative Analysis between Ultrasonometry and Computer-Aided Tomography to Evaluate Bone Healing

An ultrasonometric and computed-tomographic study of bone healing was undertaken using a model of a transverse mid-shaft osteotomy of sheep tibiae fixed with a semi-flexible external fixator. Fourteen sheep were operated and divided into two groups of seven according to osteotomy type, either regular or by segmental resection. The animals were killed on the 90th postoperative day and the tibiae resected for the in vitro direct contact transverse and axial measurement of ultrasound propagation velocity (UV) followed by quantitative computer-aided tomography (callus density and volume) through the osteotomy site. The intact left tibiae were used for control, being examined in a symmetrical diaphyseal segment. Regular osteotomies healed with a smaller and more mature callus than resection osteotomies. Axial UV was consistently and significantly higher (p?=?0.01) than transverse UV and both transverse and axial UV were significantly higher for the regular than for the segmental resection osteotomy. Transverse UV did not differ significantly between the intact and operated tibiae (p?=?0.20 for regular osteotomy; p?=?0.02 for resection osteotomy), but axial UV was significantly higher for the intact tibiae. Tomographic callus density was significantly higher for the regular than for the resection osteotomy and higher than both for the intact tibiae, presenting a strong positive correlation with UV. Callus volume presented an opposite behavior, with a negative correlation with UV. We conclude that UV is at least as precise as quantitative tomography for providing information about the healing state of both regular and resection osteotomy. (C) 2011 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:10761082, 2012

Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Computer-assisted design and manufacture of implants in the late reconstruction of extensive orbital fractures

To evaluate the use of computer-assisted designed and manufactured (CAD/CAM) orbital wall and floor implants for late reconstruction of extensive orbital fractures.