Use of the Normalized Absorbance Ratio as an Internal Standardization Approach To Minimize Measurement Error in Enzyme-Linked Immunosorbent Assays for Diagnosis of Human Papillomavirus Infection

The serological detection of antibodies against human papillomavirus (HPV) antigens is a useful tool to determine exposure to genital HPV infection and in predicting the risk of infection persistence and associated lesions. Enzyme-linked immunosorbent assays (ELISAs) are commonly used for seroepidemiological studies of HPV infection but are not standardized. Intra-and interassay performance variation is difficult to control, especially in cohort studies that require the testing of specimens over extended periods. We propose the use of normalized absorbance ratios (NARs) as a standardization procedure to control for such variations and minimize measurement error. We compared NAR and ELISA optical density (OD) values for the strength of the correlation between serological results for paired visits 4 months apart and HPV-16 DNA positivity in cervical specimens from a cohort investigation of 2,048 women tested with an ELISA using HPV-16 virus-like particles. NARs were calculated by dividing the mean blank-subtracted (net) ODs by the equivalent values of a control serum pool included in the same plate in triplicate, using different dilutions. Stronger correlations were observed with NAR values than with net ODs at every dilution, with an overall reduction in nonexplained regression variability of 39%. Using logistic regression, the ranges of odds ratios of HPV-16 DNA positivity contrasting upper and lower quintiles at different dilutions and their averages were 4.73 to 5.47 for NARs and 2.78 to 3.28 for net ODs, with corresponding significant improvements in seroreactivity-risk trends across quintiles when NARs were used. The NAR standardization is a simple procedure to reduce measurement error in seroepidemiological studies of HPV infection.

Techniques in electromanometry of the upper esophageal sphincter

The various pull-through techniques used to measure pressure and length of the upper esophageal sphincter have not been studied in depth. Because of the importance of obtaining further information about these techniques, a group of 25 dogs was submitted to upper esophageal sphincter pressure and length measurements by three different pull-through techniques (two of the continuous type and one of the station type), each with three replications per animal, to determine possible differences among techniques. A single introduction provided reliable results, with no sequential effect of the measurements or of any one technique on resting pressure. However, each technique yielded different sphincter lengths, higher values being obtained when the continuous techniques were used.

Cutaneous hypersensitivity induced in dogs and guinea-pigs by extracts of the tick Rhipicephalus sanguineus (Acari: Ixodidae)

The cutaneous hypersensitivity induced by Rhipicephalus sanguineus tick extract in dogs (natural host) and guinea-pigs (laboratory host) was evaluated. The left ear of infested and control (tick-bite naive) dogs and guinea-pigs was injected intradermally with an extract from unfed adult ticks and the right ear with phosphate buffered saline (PBS). Ear thickness variations were then measured after 10 min and 1, 2, 6, 18, 24, 48, 72 and 96 h post-injection. Results were expressed as percentual changes in the ear thickness in relation to pre-inoculation values. The final variation in ear thickness induced by the extract was given by subtracting, in each animal, the right ear percentual increase from that of the left ear. Guinea-pigs were tested at two different times following infestation and with two different doses of extract. Infested guinea-pigs from the three experiments developed an immediate (within the first 2 h post-inoculation) and a strong delayed reaction (24 h) to the extract. Dogs, unlike guinea-pigs, developed only a strong immediate reaction whereby an 80% increase in ear thickness was observed. Control animals, with the exception of one dog, did not develop any significant reaction to the extract. Only mild reactions were induced by PBS in the right ear of all animals. The correlation between the absence of a strong delayed type reaction to tick extract and the lack of resistance of the natural host to R. sanguineus tick is discussed. © 1995 Chapman & Hall.

Cloning and expression of the cDNA encoding rat granulocyte colony-stimulating factor

Granulocyte colony-stimulating factor (G-CSF) acts on precursor hematopoietic cells to control the production and maintenance of neutrophils. Recombinant G-CSF (re-G-CSF)is used clinically to treat patients with neutropenia and has greatly reduced the infection risk associated with bone marrow transplantation. Cyclic hematopoiesis, a stem cell defect characterized by severe recurrent neutropenia, occurs in man and grey collie dogs, and can be treated by administration of re-G-CSF. Availability of the rat G-CSF cDNA would benefit the use of rats as models of gene therapy for the treatment of cyclic hematopoiesis. In preliminary rat experiments, retroviral-mediated expression of canine G-CSF caused neutralizing antibody formation which precluded long-term increases in neutrophil counts. To overcome this problem we cloned the rat G-CSF cDNA from RNA isolated from skin fibroblasts. The rat G-CSF sequence shared a high degree of identity in both the coding and non-coding regions with both the murine G-CSF (85%) and human G-CSF (74%). The signal peptides of murine and human G-CSF both contained 30 amino acids (aa), whereas the deduced signal sequence for rat G-CSF possessed 21 aa. A retrovirus encoding the rat G-CSF cDNA synthesized bioactive G-CSF from transduced vascular smooth muscle cells.

Investigation into the use of the Laryngeal Mask Airway in pentobarbital anesthetized dogs

Objective - To investigate the use of the laryngeal mask airway (LMA) in dogs. Study Design - Prospective experimental study. Animals - Eight healthy adult mixed breed dogs weighing from 15 to 20 kg. Methods - The dogs were anesthetized with intravenous pentobarbital. An LMA was introduced after the induction of anesthesia and 1 L/min O2 plus 1 L/min air was delivered using a circle anesthetic system. Respiratory rate, tidal volume, arterial O2 saturation (pulse oximetry), end tidal CO2, inspired fraction of O2, pulse rate, and mean arterial blood pressure were measured after the insertion of the LMA and 30, 60, 90, and 120 minutes afterwards. Results - There were no changes in respiratory rate, tidal volume, arterial O2 saturation, and pulse rate during anesthesia. End tidal CO2 decreased significantly by the end of anesthesia and ventilation appeared satisfactory. Conclusions - An LMA appeared to be an alternative option to maintain the patency of the airway in dogs. Clinical Relevance - This device may allow safe maintenance of an airway in dogs when intubation is difficult or when it interferes with the procedure (eg, cervical myelography). ©Copyright 1999 by The American College of Veterinary Surgeons.

Tracheobronchial consequences of the use of heat and moisture exchangers in dogs

Purpose: To determine the effect of heat and moisture exchange (HME) on the tracheobronchial tree (TBT) using a unidirectional anesthesic circuit with or without CO2 absorber and high or low fresh gas flow (FGF), in dogs. Methods: Thirty-two dogs were randomly allocated to four groups: G1 (n = 8) valvular circuit without CO2 absorber and high FGF (5 L·min-1); G2 (n = 8) as G1 with HME; G3 (n = 8) circuit with CO2 absorber with a low FGF (1 L·min-1); G4 (n = 8) as G3 with HME. Anesthesia was induced and maintained with pentobarbital. Tympanic temperature (TT), inhaled gas temperature (IGT), relative (RH) and absolute humidity (AH) of inhaled gas were measured at 15 (control), 60, 120 and 180 min of controlled ventilation. Dogs were euthanized and biopsies in the areas of TBT were performed by scanning electron microscopy. Results: The G2 and G4 groups showed the highest AH (>20 mgH2O·L-1) and G1 the lowest (< 10 mgH2O·L-1) and G3 was intermediate (<20 mgH2O·L-1) (P < 0.01). There was no difference of TT and IGT among groups. Alterations of the mucociliary system were greatest in G1, least in G2 and G4, and intermediate in G3. Conclusion: In dogs, introduction of HME to a unidirectional anesthetic circuit with/without CO2 absorber and high or low FGF preserved humidity of inspired gases. HME attenuated but did not prevent alterations of the mucociliary system of the TBT.

Reduction of diameter of the left ventricle of dogs by plication of the left ventricular free wall

Objective - To determine effects of reducing the diameter of the left ventricle of dogs by plication of the left ventricular free wall. Animals - 8 healthy adult mixed-breed dogs. Procedure - Left lateral thoracotomy and a T-shaped pericardiotomy were performed. The free wall of the left ventricle was imbricated with 3 interrupted transfixing sutures applied in a horizontal mattress pattern, using 3-0 polypropylene suture assembled on a straight cutting needle. Surgeons were careful to avoid the coronary vessels. Echocardiography was performed 24 hours before and 48 hours after surgery. Electrocardiography was performed before and 1, 2, 7, 15, 21, 30, and 60 days after surgery. Results - Echocardiographic measurements revealed that the diameter of the left ventricle was reduced by a mean of 23.5%. Electrocardiography revealed ventricular premature complexes 24 hours after surgery that regressed without treatment during the first week after surgery. Conclusions and Clinical Relevance - Plication of the left ventricular free wall of dogs can reduce end-diastolic and end-systolic dimensions of the left ventricle. The technique is simple and does not require cardiopulmonary bypass. According to Laplace's law, the reduction of cardiac diameter leads to reduction on free-wall tension and may improve left ventricular function in dilatated hearts. Thus, additional studies involving dogs with dilated cardiomyopathy should be conducted.

Non-Specific Interaction between Ants (Hymenoptera; Formicidae) and Fruits of Syagrus romanzoffiana (Arecaceae) in an Area of the Brazilian Atlantic Forest

On the forest floor of a fragment of the Atlantic Forest which is considered to be in a stage of recuperation, located in southeastern Brazil, we recorded 54 species of ants interacting with the fruits of Syagrus romanzoffiana (Arecaceae). Myrmicinae was the subfamily which was the richest in species, followed by Ponerinae, Formicinae, Ecitoninae, Dolichoderinae, Pseudomyrmecinae and Cerapachyinae. The fruits of S. romanzoffiana can be classified according to whether or not they are preyed by the squirrel Sciurus ingrami, and the ant fauna that visits the fruits has an average similarity. Regarding diversity, the data suggest that there is no significant difference between ant assemblages at preyed and non-preyed-upon fruit falls. The results also indicate that there is no specific interaction between the ant fauna that visits the fruits of S. romanzoffiana, as it consists largely of species with omnivorous feeding habits.

Plication of the free wall of the left ventricle in dogs with doxorubicin-induced cardiomyopathy

Objective - To evaluate plication of the free wall of the left ventricle, which reduces the left ventricular area and volume, as a method to improve the left ventricular systolic function without cardiopulmonary bypass. Animals - 8 mixed-breed adult dogs. Procedure - Dilated cardiomyopathy (DCM) was induced in each dog by administration of doxorubicin (30 mg/m2, IV, q 21 d for 168 days). Two dogs died during induction of cardiomyopathy. Plication surgery was performed in 4 dogs. Two dogs did not ondergo to surgery (control group). Values for cardiac output (CO), 2-dimensional and M-mode echocardiography, arterial blood pressure, electrocardiography, blood cell counts, and serum biochemical analyses were recorded after induction of DCM (baseline) and 1, 2, 7, 15, 21, 30, 60, 90, 120, 150, and 180 days after plication surgery. Ambulatory ECG (Holter) recordings were conducted for 24 hours on the day of surgery. Results - 1 dog died after plication surgery. The remaining dogs undergoing ventricular plication had a significant improvement in CO, ejection fraction, and fractional shortening and reductions of left ventricular area and volume after surgery. Electrocardiographic and Holter recordings revealed premature ventricular complexes, which resolved without treatment during the first week after surgery. Clinical condition of the control dogs declined, and these 2 dogs died approximately 40 days after induction of cardiomyopathy. Conclusions and Clinical Relevance - Plication of the free wall of the left ventricle improved left ventricular systolic function in dogs with doxorubicin-induced cardiomyopathy. Additional studies are needed to evaluate its application in dogs with naturally developing DCM.

Effect of rotary instrumentation and of the association of calcium hydroxide and chlorhexidine on the antisepsis of the root canal system in dogs

This study aimed at evaluating the antisepsis of the root canal system (RCS) and periapical region (PR) provided by rotary instrumentation associated with chlorhexidine + calcium hydroxide as intracanal medicament. Chronic periapical lesions were induced in 26 pre-molar roots in two dogs. After microbiological sampling, automatic instrumentation using the Profile system and irrigation with 5.25% sodium hypochlorite solution, with a final rinse of 14.3% EDTA followed by profuse irrigation with physiological saline were carried out in 18 root canals. After drying the canals, a paste based on calcium hydroxide associated with a 2% chlorhexidine digluconate solution was placed inside them. After 21 days, the medication was removed, leaving the root canals empty and coronally sealed. After 96 hours, a final microbiological sample was obtained, followed by histomicrobiological processing by the Brown & Brenn method. Eight untreated root canals represented the control group (C-G). Based on the Mann-Whitney test at a confidence level of 5% (p < 0.05), the procedures of antisepsis used offered significant efficacy (p < 0.05) resulting in 100.0% of the canals free of microorganisms. In the C-G, an elevated incidence of various microbial morphotypes was confirmed in all sites of the RCS, with the presence of microbial colonies in the periapical region. In contrast, the experimental group showed a similar pattern of infection in the RCS, although less intense and a reduced level of periapical infection (p < 0.05). It was concluded that adequate instrumentation followed by the application of calcium hydroxide + chlorhexidine offered significant elimination of microorganisms.

Serological profile of anti-Toxoplasma gondii antibodies in apparently healthy dogs of the city of Botucatu, São Paulo state, Brazil

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, a coccidian protozoan of worldwide distribution. The seroprevalence in canine population can be an alternative for measuring T. gondii urban spreading. A total of 780 blood samples from dogs were collected, during the yearly anti-rabies campaign, carried out by the Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine and Animal Husbandry (FMVZ), São Paulo State University, UNESP, together with the county health authorities, in August 1999. Using Indirect Fluorescent Antibody Test (IFAT) for detecting antibodies anti-T. gondii in the sera samples, we observed that 258 dogs (33.1%) were positive. The associations between the serological results and the epidemiological variables were studied. Statistically significant differences were not found regarding sex (32.2% male and 34.3% female reactors). Dogs without a defined breed showed seropositivity statistically higher than the pedigreed group. The occurrence of infection was considered higher with age.

Carotid arteries in the dog: Structure and histophysiology

Light microscopy analysis of the mural structure of the common carotid artery, internal carotid artery and external carotid artery in mongrel dog revealed variability in the middle values of vascular diameter and thickness of the intimal plus medial coats and adventitial coat. The vascular diameter did not differ between the internal carotid artery and external carotid artery, but was significantly increased in the common carotid artery from which the other two arteries had origin. The increased thickness of the intimal plus medial coats of the common carotid artery revealed the constant mechanical adjustment of the arterial wall and local shearing stress that occurred among the arterial coats. The adventitial coat of the internal carotid artery was significantly thicker than those of the common carotid and external carotid arteries, with no significant difference noted between the latter arteries. These histomorphometric results were related to the qualitative observations regarding the mural structure of the three arteries analyzed, especially regarding to the increased thickness and structural complexity of the adventitial coat of the internal carotid artery. Perhaps it acted as an external protective sheath of the vascular wall during its long course until the carotid channel. © 2006 Sociedad Chilena de Anatomía.

Proposal of a new technique for bile duct reconstruction after iatrogenic injury. Study in dogs and review of the literature

Purpose: Interposition of a jejunal tube between the common bile duct and duodenum. Methods: Five adult mongrel dogs of both sexes, weighing on average 22.3 kg (18 to 26.5 kg), were used. Obstructive jaundice was induced by ligation of the distal common bile duct. After one week, a 2.5-cm long jejunal tube was fabricated from a segment of the loop removed 15 cm from the Treitz angle and interposed between the common bile duct and duodenum. Results: The animals presented good clinical evolution and no complications were observed. After 6 weeks, complete integration was noted between the bile duct mucosa, tube and duodenum and a significant reduction in total bilirubin and alkaline phosphatase was observed when compared to the values obtained one week after ligation of the common bile duct. Conclusion: The jejunal tube interposed between the dilated bile duct and duodenum showed good anatomic integration and reduced total bilirubin and alkaline phosphatase levels in the animals studied.

Response of the stingless bee manduri do mato grosso (Melipona favosa orbignyi, Hymenoptera, Apidae) to variable concentrations of sucrose in artificial feeding sources of different colors

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