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Biomedical subjects

M J Silva

Publications and source records attributed to M J Silva.

At least 37 records · Page 2Linked to original sources

Intrasynovial flexor tendon repair. An experimental study comparing low and high levels of in vivo force during rehabilitation in canines.

BACKGROUND: Rehabilitation methods that generate increased tendon force and motion have been advocated to improve results following intrasynovial flexor tendon repair. However, the effects of rehabilitation force and motion on tendon-healing may be masked by the high stiffness produced by newer suture methods. Our objective was to determine whether the biomechanical properties of tendons repaired by one of two multistrand suture methods were sensitive to an increased level of applied rehabilitation force. METHODS: Two hundred and fourteen flexor digitorum profundus tendons from 107 adult dogs were transected and repaired. Dogs were assigned to one of four groups based on the rehabilitation method (low force [<5 N] or high force [17 N]) and the repair technique (four-strand or eight-strand core suture) and were killed between five and forty-two days after the procedure. Repair-site structural properties were determined by tensile testing, and digital range of motion was assessed with use of a motion-analysis system. RESULTS: Tensile properties did not differ between the low and high-force rehabilitation groups, regardless of the repair technique (p > 0.05). In contrast, tensile properties were strongly affected by the repair technique, with tendons in the eight-strand group having an approximately 35% increase in ultimate force and rigidity compared with those in the four-strand group (p < 0.05). Ultimate force did not change significantly with time during the first twenty-one days (p > 0.05); there was no evidence of softening in either of the repair or rehabilitation groups. Force increased significantly from twenty-one to forty-two days, while rigidity increased throughout the forty-two-day period (p < 0.05). CONCLUSIONS: Increasing the level of force applied during postoperative rehabilitation from 5 to 17 N did not accelerate the time-dependent accrual of stiffness or strength. Suture technique was of primary importance in providing a stiff and strong repair throughout the early healing interval. CLINICAL RELEVANCE: Our findings suggest that there be a reexamination of the concept that increases in force produced by more vigorous mobilization protocols are beneficial to tendon-healing. While more vigorous rehabilitation may help to improve hand function, we found no evidence that it enhances tissue-healing or strength in the context of a modern suture repair.

Animals↗

Quantitative detection of eight phthalate metabolites in human urine using HPLC-APCI-MS/MS.

Because of the ubiquity of phthalates and their potential role in increasing risk for cancer and reproductive dysfunction, the need for human exposure assessment studies is urgent. In response to this need, we developed a high-throughput, robust, sensitive, accurate, and precise assay for simultaneous measurement of trace levels of eight phthalate metabolites in human urine by HPLC-MS/MS. Human urine samples were processed using enzymatic deconjugation of the glucuronides followed by solid-phase extraction. The eluate was concentrated, and the phthalate metabolites were chromatographically resolved by reversed-phase HPLC, detected by APCI-tandem mass spectrometry, and quantified by isotope dilution. This selective analytical method permits rapid detection (7.7 min total run time) of eight urinary metabolites of the most commonly used phthalates with detection limits in the low nanagram per milliliter range. Assay precision was improved by incorporating 13C4-labeled internal standards for each of the eight analytes, as well as a conjugated internal standard to monitor deconjugation efficiency. This selective, sensitive, and rapid method will help elucidate potential associations (if any) between human exposure to phthalates and adverse health effects.

Calibration↗

Effects of the water soluble fraction from leaves of Ageratum conyzoides on smooth muscle.

The water soluble fraction of Ageratum conyzoides L. (WSF) was studied in isolated rat uterus and intestinal smooth muscles in order to evaluate its popular use as a spasmolytic. WSF (0.2 and 0.4 mg/mL) increased EC(50) values and decreased maximum responses to acetylcholine and calcium chloride. WSF (0.5-3.3 mg/mL) also produced direct myorelaxant effect on smooth muscle preparations. Theophylline (10(-3) M) potentiated the relaxant action of WSF. Theophylline also prevented the decrease in maximum response promoted by WSF in acetylcholine concentration-effect curves. These results seem to be partially linked to calcium mobilization. The data also suggest that WSF could act synergistically with theophylline in the inhibition of cyclic AMP phosphodiesterase. The results give support to the popular medicinal indications of the plant.

Acetylcholine↗

In vitro sodium fluoride exposure decreases torsional and bending strength and increases ductility of mouse femora.

Fluoride exposure in vivo can reduce the material strength of bone, an effect that has been attributed to a change in mineral structure. An in vitro model of fluoride exposure offers the potential to study directly the effects of fluoride on bone mineral. Previous investigators have reported that soaking bones in sodium fluoride in vitro reduces bone strength. However, long soaking times and the absence of physiological buffering ions from their treatment solutions may have caused mineral dissolution that contributed to the decrease in bone strength. Our objectives were to further characterize the effects of in vitro fluoride exposure on bone mechanical properties and to determine if the changes reported in previous studies of bovine cortical bone would be observed for whole rodent bones. We soaked 60 mouse femora in sodium fluoride solutions, with and without physiological buffering ions, and evaluated their torsional and bending properties. Fluoride soaked bones had a 30-fold increase in fluoride content and a 23% increase in water content compared to controls. These changes were associated with average reductions in ultimate load of 45%, reductions in rigidity of 70%, and increases in deformation to failure of 80%. The effect of fluoride was similar for bones treated in buffered and non-buffered solutions, and was observed in both torsion and bending. Our findings confirm those of previous studies and highlight the strong effect that in vitro fluoride exposure has on bone mechanical properties. The in vitro model of fluoride exposure offers a tool to further study the effects of ion substitution in bone.

Animals↗

The resistance of a four- and eight-strand suture technique to gap formation during tensile testing: an experimental study of repaired canine flexor tendons after 10 days of in vivo healing.

There is a high incidence of gap formation at the repair site following tendon repair. Our goal was to determine the resistance of a 4- and an 8-strand suture technique to gap formation during tensile testing. We hypothesized that the 8-strand repair would sustain higher force levels at the onset of 1- and 3-mm gaps than the 4- strand repair. Twenty-two canine flexor tendons were transected, repaired, and tested to failure after 10 days of in vivo healing. Tests were recorded using a 60-Hz video system that allowed frame-by-frame playback for assessment of gap formation. The 8-strand repairs sustained 80% higher force at a gap of 1 mm than the 4-strand repairs (average force, 70 vs 39 N), but the force sustained at a gap of 3 mm did not differ between groups (35 N for both groups). For both repair types, a 1-mm gap typically occurred near the point of ultimate (maximum) force while a 3-mm gap occurred after the ultimate force. We conclude that the 8-strand repair is significantly more resistant to initial gapping during ex vivo tensile testing than the 4-strand repair but that the two repairs are equally susceptible to rupture if a gap of 3 mm or greater forms.

Animals↗

A new species of Calomys (Rodentia, Sigmodontinae) from Central Brazil identified by its karyotype.

Ten species of small rodents of genus Calomys are found in South America. Three of these ten species are known to occur in Brazil: C. tener, C. laucha and C. expulsus (= C. callosus expulsus). Almost all Calomys karyotypes are made up of acrocentric pairs. In this paper we describe a new karyotype with 2n = 46 (FN = 66), including 11 meta/submetacentric and 11 acrocentric autosomal pairs. This is not related to any described Calomys karyotype. The X chromosome is a medium submetacentric and the Y is a small acrocentric. This new karyotype is briefly compared to karyotype of the sympatric species C. tener (2n = 66, FN = 66). The reduced diploid number and small amount of pericentromeric heterochromatin observed in the biarmed chromosomes that contrasts to large blocks seen in acrocentrics seem to indicate that centric fusion and loss of constitutive heterochromatin have led to the new karyotype. Cytogenetic evidence suggests strongly that a new species with 2n = 46 from Central Brazil should be described in the genus Calomys.

Animals↗

Levels of seven urinary phthalate metabolites in a human reference population.

Using a novel and highly selective technique, we measured monoester metabolites of seven commonly used phthalates in urine samples from a reference population of 289 adult humans. This analytical approach allowed us to directly measure the individual phthalate metabolites responsible for the animal reproductive and developmental toxicity while avoiding contamination from the ubiquitous parent compounds. The monoesters with the highest urinary levels found were monoethyl phthalate (95th percentile, 3,750 ppb, 2,610 microg/g creatinine), monobutyl phthalate (95th percentile, 294 ppb, 162 microg/g creatinine), and monobenzyl phthalate (95th percentile, 137 ppb, 92 microg/g creatinine), reflecting exposure to diethyl phthalate, dibutyl phthalate, and benzyl butyl phthalate. Women of reproductive age (20-40 years) were found to have significantly higher levels of monobutyl phthalate, a reproductive and developmental toxicant in rodents, than other age/gender groups (p < 0.005). Current scientific and regulatory attention on phthalates has focused almost exclusively on health risks from exposure to only two phthalates, di-(2-ethylhexyl) phthalate and di-isononyl phthalate. Our findings strongly suggest that health-risk assessments for phthalate exposure in humans should include diethyl, dibutyl, and benzyl butyl phthalates.

Adult↗

Increased levels of sister chromatid exchanges in military aircraft pilots.

Sister chromatid exchanges (SCEs) were scored in lymphocytes of nine high-performance pilots of alphajet aircrafts and of ten control individuals from the same air base. Statistical analysis of the mean SCE count per cell in the total number of cells analyzed as well as in those having 12 or more SCEs (high-frequency cells, HFCs) revealed a significant difference between pilots and controls, after adjusting for the effect of smoking. Analysis of the cell cycle kinetic data (replication and mitotic indices) revealed no significant differences either between pilots and controls or between smokers and nonsmokers. Previously, we reported an increase in the SCE levels in workers of the aeronautical industry exposed to noise and whole-body vibration. The present results corroborate those findings and indicate that noise and whole-body vibration may cause genotoxic effects in man.

Adult↗

Reduced bone stress as predicted by composite beam theory correlates with cortical bone loss following cemented total hip arthroplasty.

Clinical and experimental evidence suggest that periprosthetic bone loss following total hip arthroplasty is caused in part by stress-shielding. Changes in bone stress in the proximal femur following implantation can be estimated with use of composite beam theory. We hypothesized that the degree of stress-shielding predicted by beam theory correlates with the magnitude of bone loss following cemented total hip arthroplasty. We analyzed cross sections from the proximal femur of 13 patients who had undergone unilateral cemented total hip arthroplasty. A matching implant was inserted contralaterally, and the cross-sectional properties of the implant and bone and the bone density were determined. Bone loss was calculated on the basis of differences between contralateral (control) and ipsilateral (remodeled) sections and correlated to several beam-theory parameters calculated from the control sections: implant rigidity, bone rigidity, ratio of implant to bone rigidity, and predicted decrease in bone stress. All parameters except implant rigidity were significantly correlated with bone loss (p < 0.05). Parameters that included implant and bone properties were more strongly correlated with bone loss than were those based on bone properties alone. The predicted decrease in bone stress explained 50-60% of the variance in bone loss. The data also indicated that patients were not likely to lose substantial amounts of bone unless the reduction in bone stress exceeded a threshold value. Although limited by a small and heterogeneous sample, these results indicate that beam-theory predictions correlate with the degree of femoral resorption and should be investigated further as a means to identify patients at high risk for bone loss.

Arthroplasty, Replacement, Hip↗

Effects of increased in vivo excursion on digital range of motion and tendon strength following flexor tendon repair.

Postoperative rehabilitation is an important factor in determining functional outcome following intrasynovial flexor tendon repair. We hypothesized that a rehabilitation protocol that produced increased in vivo excursion would lead to increased digital range of motion and tendon strength and decreased adhesion formation in a canine model. Ninety-six flexor digitorum profundus tendons from 48 dogs were cut transversely and repaired by a multistrand suture technique. Postoperative rehabilitation was performed daily with a low excursion-low force (1.7-mm average excursion; < 10 N force) or a high excursion-low force (3.6 mm excursion; < 10 N force) protocol. After death of the dogs at 10, 21, or 42 days, specimens were evaluated for digital range of motion, tensile mechanical properties, elongation of the repair site, and adhesion formation. Our data indicate that the range of motion of digits whose tendons were at low or high excursion was similar to that of controls. Increased in vivo tendon excursion due to synergistic wrist motion did not significantly affect ex vivo flexion of the distal and proximal interphalangeal joints or tendon displacement (p > 0.05). Similarly, tensile properties (ultimate load, repair site rigidity, and repair site strain at 20 N and at failure) and length of the gap at the repair site were not significantly affected by increased excursion (p > 0.05). Severity of adhesion formation was reduced slightly by increased excursion (p = 0.04). Our findings indicate that 1.7 mm of tendon excursion is sufficient to prevent adhesion formation following sharp transection of the canine flexor tendon and that additional excursion provides little added benefit.

Animals↗

Wrist and digital joint motion produce unique flexor tendon force and excursion in the canine forelimb.

The force and excursion within the canine digital flexor tendons were measured during passive joint manipulations that simulate those used during rehabilitation after flexor tendon repair and during active muscle contraction, simulating the active rehabilitation protocol. Tendon force was measured using a small buckle placed upon the tendon while excursion was measured using a suture marker and video analysis method. Passive finger motion imposed with the wrist flexed resulted in dramatically lower tendon force (approximately 5 N) compared to passive motion imposed with the wrist extended (approximately 17 N). Lower excursions were seen at the level of the proximal interphalangeal joint with the wrist flexed (approximately 1.5 mm) while high excursion was observed when the wrist was extended or when synergistic finger and wrist motion were imposed (approximately 3.5 mm). Bivariate discriminant analysis of both force and excursion data revealed a natural clustering of the data into three general mechanical paradigms. With the wrist extended and with either one finger or four fingers manipulated, tendons experienced high loads of approximately 1500 g and high excursions of approximately 3.5 mm. In contrast, the same manipulations performed with the wrist flexed resulted in low tendon forces (4-8 N) and low tendon excursions of approximately 1.5 mm. Synergistic wrist and finger manipulation provided the third paradigm where tendon force was relatively low (approximately 4 N) but excursion was as high as those seen in the groups which were manipulated with the wrist extended. Active muscle contraction produced a modest tendon excursion (approximately 1 mm) and high or low tendon force with the wrist extended or flexed, respectively. These data provide the basis for experimentally testable hypotheses with regard to the factors that most significantly affect functional recovery after digital flexor tendon injury and define the normal mechanical operating characteristics of these tendons.

Animals↗

Sensitivity of Fusarium strains to Chelidonium majus L. extracts.

Ten Fusarium strains were tested for their sensitivity to extracts of Chelidonium majus L. Growth inhibition was measured either in solid or in liquid media. Aqueous extracts had considerable inhibitory action but methanolic extracts showed the best results. Root extracts were more inhibitory than shoot extracts. On the basis of growth inhibition the Fusarium strains were aggregated into five classes, the extremes being Fusarium culmorum plus Fusarium graminearum (quite resistant) and Fusarium oxysporum f. sp. cubense (very sensitive), with the other seven strains occupying the three intermediate classes. The high resistance of most Fusarium strains to conventional fungicides led us to propose C. majus as a good source of substances useful for the treatment of fungal infections, with special importance for those caused by Fusarium.

Antifungal Agents↗

Autosomal and sex chromosomal polymorphisms with multiple rearrangements and a new karyotype in the genus Rhipidomys (Sigmodontinae, Rodentia).

Two diploid numbers and five karyomorphs were found in ten specimens of Rhipidomys (Sigmodontinae, Rodentia) from three states in Brazil: 2n = 50 from Amazonas, and 2n = 44 from Mato Grosso and Bahia. CBG, GTG, and RBG-banding and Ag-NOR analyses were performed, as well as fluorescence in situ: hybridization with (T2AG3)7 probes. The new diploid number of 2n = 50 was associated with two different fundamental numbers (FN = 71 and 72) as a result of pericentric inversions and addition/deletion of constitutive heterochromatin. The samples from two localities (Aripuanã and Vila Rica) in the state of Mato Grosso shared 2n = 44 and FN = 52, but their karyotypes differed because of pericentric inversions. Although the single specimen from Bahia had the same diploid number as the samples from Mato Grosso, its karyotype and FN were completely distinctive. Karyological comparison of GTG-banding patterns revealed total homology between the karyotypes of the specimens from Bahia and Mato Grosso, implying the occurrence of 14 autosomal pericentric inversions. Homologies between ten of the autosomes in the karyotypes with 2n = 50 (FN = 72) and 2n = 44 (from Vila Rica, MT) were demonstrated. The differentiation between 2n = 44 and 2n = 50 involved five pericentric inversions, addition/deletion of constitutive heterochromatin in both autosomes and sex chromosomes, at least one Robertsonian rearrangement and other not detected rearrangements. Despite the remarkable number of rearrangements, interstitial telomeric sites (ITS) were not detected. Sex chromosomes also exhibited polymorphism in size and morphology.

Animals↗

Growing C57Bl/6 mice increase whole bone mechanical properties by increasing geometric and material properties.

In vivo murine models are becoming increasingly important in bone research. To establish baseline data for researchers using these models, we studied the long bones from C57BL/6 female mice, a strain that is widely used in bone research. We determined the femoral structural and material properties in both torsion and four-point bending for mice at ages 4-24 weeks. Measurements of femoral cross-sectional geometry and tibial densitometric properties were also obtained. Results indicated that all structural properties (except ultimate energy), changed significantly with age (p < 0.001). Ultimate torque, ultimate moment, torsional rigidity, and bending rigidity all increased to peak values at 20 weeks, whereas ultimate rotation and ultimate displacement decreased to minimum values at 20 weeks. Our data indicate that increases in the material properties contributed more than increases in cross-sectional geometry to the changes in structural rigidity and ultimate load. For example, from 4-20 weeks torsional rigidity increased 1030%, while shear modulus increased 610% and the polar moment of inertia (a measure of the geometric resistance to rotation) increased by only 85%. Changes in the cross-sectional geometry with age were due to increases in periosteal diameter and decreases in endosteal diameter. In general, both torsion and bending techniques revealed large changes in structural and material properties with age. We conclude that peak bone strength is not achieved before 20 weeks in C57BL/6 female mice, and that torsion and four-point bending tests are equally well suited for evaluating mechanical properties of murine long bones.

Age Factors↗

The effect of gap formation at the repair site on the strength and excursion of intrasynovial flexor tendons. An experimental study on the early stages of tendon-healing in dogs.

BACKGROUND: Elongation (gap formation) at the repair site has been associated with the formation of adhesions and a poor functional outcome after repair of flexor tendons. Our objectives were to evaluate the prevalence of gap formation in a clinically relevant canine model and to assess the effect of gap size on the range of motion of the digits and the mechanical properties of the tendons. METHODS: We performed operative repairs after sharp transection of sixty-four flexor tendons in thirty-two adult dogs. Rehabilitation with passive motion was performed daily until the dogs were killed at ten, twenty-one, or forty-two days postoperatively. Eight tendons ruptured in vivo. In the fifty-six intact specimens, the change in the angles of the proximal and distal interphalangeal joints and the linear excursion of the flexor tendon were measured as a 1.5-newton force was applied to the tendon. The gap at the repair site was then measured, and the isolated tendons were tested to failure in tension. RESULTS: Twenty-nine tendons had a gap of less than one millimeter, twelve had a gap of one to three millimeters, and fifteen had a gap of more than three millimeters. Neither the time after the repair nor the size of the gap was found to have a significant effect on motion parameters (p > 0.05); however, the ultimate force, repair-site rigidity, and repair-site strain at twenty newtons were significantly affected by these parameters (p < 0.05). Testing of the tendons with a gap of three millimeters or less revealed that, compared with the ten-day specimens, the forty-two-day specimens failed at a significantly (90 percent) higher force (p < 0.01) and had a significantly (320 percent) increased rigidity (p < 0.01) and a significantly (60 percent) decreased strain at twenty newtons (p < 0.05). In contrast, the tensile properties of the tendons that had a gap of more than three millimeters did not change significantly with time. CONCLUSIONS: Our data indicate that, in a dog model involving sharp transection followed by repair, a gap at the repair site of more than three millimeters does not increase the prevalence of adhesions or impair the range of motion but does prevent the accrual of strength and stiffness that normally occurs with time.

Animals↗

[Anaphylaxis and allergic reactions]

OBJECTIVE: To review the etiologic factors, pathophysiology and specially the therapeutic management of the anaphylactic reactions.METHODS: The bibliographic review included textbooks of allergy and intensive care and Medline references.RESULTS: Anaphylactic reactions that result from the rapid release of potent pharmacologically active mediators from mast cells or peripheral blood basophils may involve several organ systems with some risk of death.COMMENTS: Anaphylaxis is a clinical syndrome with significant mortality rate. There are several etiologic factors that trigger the process and multiple pathophysiologic mechanisms involved. Early diagnosis and an effective therapy lead to a better prognosis. In the majority of the cases prophylactic measures are needed to avoid other episodes.

Journal Article↗

Sister chromatid exchange analysis in workers exposed to noise and vibration.

BACKGROUND: There has been a growing interest in the combined effects of noise and vibration. In a population of aeronautical workers diagnosed with vibroacoustic disease (VAD), a large incidence of malignancy was detected. These workers were exposed to large pressure amplitude (LPA) (> or = 90 dB SPL) noise, with energy content concentrated within the low frequency (LF) bands (< or = 500 Hz) and whole-body vibration (WBV). To our knowledge, there are no studies conducted in humans or animals that address the issue of the potential genotoxic effects of vibration combined with noise. In the present study, the levels of sister chromatid exchanges (SCE) and of cells with high frequencies of SCE (HFC) were analyzed in peripheral blood lymphocytes of workers employed in various occupations within the aeronautical industry. METHODS: SCE and HFC were analyzed in lymphocytes of 50 workers occupationally exposed to noise and vibration and of 34 office-worker controls (G0). The exposed group included: 10 hand-vibrating tool operators (G1), 15 engine test cell technicians (G2), 12 aircraft run-up technicians (G3) and 13 Portuguese Air Force helicopter pilots (G4). Groups 2-4 were exposed to WBV and LPALF noise; group 1 was exposed to LPA high frequency noise and local vibration. Statistical analysis of the mean SCE count per cell was carried out by multiple regression analysis comparing various predictor variables: type of exposure, duration of exposure, age, and cigarette consumption. RESULTS: Only cigarette consumption and type of exposure were found to be significantly correlated with the mean SCE frequency. After allowing for the effects of smoking, the analysis indicates that: 1) there was no significant difference between G1 and G0 (p > 0.05); 2) the differences between G2 and G0, G3 and G0, G4 and G0 were all highly significant (p < 0.001); 3) there was no significant difference between G2 and G3 (p > 0.05), nor between G2 and G3 combined and G4 (p > 0.05); and 4) G2 and G4 combined had a significantly elevated mean SCE frequency compared G0 (p < 0.001). Statistical analysis of the proportion of HFC was consistent with these results. CONCLUSION: Our data suggest that occupational exposure to LPALF noise and WBV may lead to increased levels of SCE in men. These results also suggest a reason for the high incidence of malignancy in VAD patients. The observed effects may not reflect a direct action of these physical agents on DNA. Alternative explanations may lie in the noise-, vibration-, and/or stress-induced pathophysiological changes.

Adult↗

Computed tomography-based finite element analysis predicts failure loads and fracture patterns for vertebral sections.

Computed tomography-based finite element analysis represents a powerful research tool for investigating the mechanics of skeletal fractures. To provide evidence that this technique can be used to predict failure loads and fracture patterns for bone structures, we compared the observed and predicted failure behaviors of 18 midsagittal sections, 10 mm thick, cut from human vertebral bodies. The specimens were scanned by computed tomography, and finite element models were generated with use of empirically determined density-property relations to assign element-specific material properties. The specimens were loaded to failure in uniaxial compression, and the models were analyzed under matching conditions. The models provided predictions of yield load that were strongly correlated with experimentally measured values (r2 > 0.86) and were typically within 25% of measured values. Predicted stiffness values were moderately correlated with measured values, but large absolute differences existed between them. Comparisons between regions of observed fracture and of high predicted strain indicated that strain was an accurate indicator of the pattern of local fracture in more than two-thirds of the bone specimens. In addition, strain contour plots provided better indicators of local fracture than did stress plots in these heterogeneous bone structures. We conclude that computed tomography-based finite element analysis can be used successfully to predict both global and local failure behavior of simplified skeletal structures.

Adult↗