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Biomedical subjects

P M Santos

Publications and source records attributed to P M Santos.

At least 19 recordsLinked to original sources

Activation and significance of vacuolar H+-ATPase in Saccharomyces cerevisiae adaptation and resistance to the herbicide 2,4-dichlorophenoxyacetic acid.

The stimulation of the activity of the H(+)-ATPase present in the vacuolar membrane (V-ATPase) of Saccharomyces cerevisiae is here described in response to a moderate stress induced by 2,4-dichlorophenoxyacetic acid (2,4-D). This in vivo activation (up to 5-fold) took place essentially during the adaptation period, preceding cell division under herbicide stress, in coordination with a marked activation of plasma membrane H(+)-ATPase (PM-ATPase) (up to 30-fold) and the decrease of intracellular and vacuolar pH values, suggesting that activation may be triggered by acidification. Single deletion of VMA1 and genes encoding other V-ATPase subunits led to a more extended period of adaptation and to slower growth under 2,4-D stress. Results suggest that a functional V-ATPase is required to counteract, more rapidly and efficiently, the dissipation of the physiological H(+)-gradient across vacuolar membrane registered during 2,4-D adaptation.

2,4-Dichlorophenoxyacetic Acid↗

New broad-host-range promoter probe vectors based on the plasmid RK2 replicon.

Broad-host-range plasmid RK2-based promoter probe vectors with a known nucleotide sequence were constructed. In the absence of an upstream promoter, the expression of two tested reporter genes (luc and lacZ) in Escherichia coli was virtually zero, while insertion of the Ptrc promoter resulted in strong inducer-dependent expression. The lacZ-based vectors were mobilized into Pseudomonas fluorescens ST, Pseudomonas putida KT2442, Sphingomonas spp. and Burkholderia spp. LB400, and expression analyses indicated that the properties observed in E. coli are maintained across the species barriers. In addition, the previously established knowledge of RK2 molecular biology allows easy manipulations of features such as plasmid copy number, further extending the application potential of the vectors.

DNA Probes↗

A functional model system of an hypoxic nerve injury and its evaluation.

OBJECTIVES/HYPOTHESIS: Develop an hypoxic peripheral nerve injury model with a controlled injury type and two types of clinically relevant physiological measurements of function during and after recovery. The model, controlling for injury and measurement variables, would have predictable outcomes in function. The functional model could test potential therapeutic interventions with greater sensitivity. STUDY DESIGN: Twenty-one rats were used in preliminary studies evaluating peroneal nerve injury types and functional model evaluation. Forty-eight rats were used in a controlled and blinded evaluation of the injury model followed by treatment with hyperbaric oxygen (HBO) as a potential therapeutic intervention and evaluated with functional models. METHODS: Preliminary studies compared nerve injuries: epineurectomy, epineurectomy with crush and transection with autograft for rate of return of function and final extent of return of function. The gait analysis model was also evaluated and modified to decrease variability. The final study evaluated peroneal epineurectomy and nerve crush injury with serial gait analysis during recovery, final elicited maximum force measurements, and histological analysis. Half of the animals were treated with HBO during recovery (ANOVA or regression statistical analysis were used to determine group differences.). RESULTS: Preliminary studies suggested that the peroneal nerve injury model of an epineurectomy with crush of specified length and a modification of the gait analysis model would yield a useful and predictable injury outcome. The final study resulted in predicted and consistent injury outcomes. In the HBO treatment group, a 12% improvement in function 5 days after HBO treatment was demonstrated (P < .03), but no long-term or histological benefit was seen. CONCLUSION: A reliable hypoxic nerve injury model has been developed and tested utilizing two functional methods as the primary outcome variables.

Animals↗

Physiological analysis of the expression of the styrene degradation gene cluster in Pseudomonas fluorescens ST.

The effects of different carbon sources on expression of the styrene catabolism genes in Pseudomonas fluorescens ST were analyzed by using a promoter probe vector, pPR9TT, which contains transcription terminators upstream and downstream of the beta-galactosidase reporter system. Expression of the promoter of the stySR operon, which codes for the styrene two-component regulatory system, was found to be constitutive and not subject to catabolite repression. This was confirmed by the results of an analysis of the stySR transcript in P. fluorescens ST cells grown on different carbon sources. The promoter of the operon of the upper pathway, designated PstyA, was induced by styrene and repressed to different extents by organic acids or carbohydrates. In particular, cells grown on succinate or lactate in the presence of styrene started to exhibit beta-galactosidase activity during the mid-exponential growth phase, before the preferred carbon sources were depleted, indicating that there is a threshold succinate and lactate concentration which allows induction of styrene catabolic genes. In contrast, cells grown on glucose, acetate, or glutamate and styrene exhibited a diauxic growth curve, and beta-galactosidase activity was detected only after the end of the exponential growth phase. In each experiment the reliability of the reporter system constructed was verified by comparing the beta-galactosidase activity and the activity of the styrene monooxygenase encoded by the first gene of the styrene catabolic operon.

Bacterial Proteins↗

Percutaneous endoscopic gastrostomy: avoiding complications.

This study evaluates our complications arising directly or indirectly from placement or management of percutaneous endoscopic gastrostomy (PEG) tubes and provides recommendations for avoidance of complications. Seventy-one patients received PEG tube placement by otolaryngologists between January 1991 and May 1997. Records were reviewed for diagnoses, combined procedures, and complications. Addressing potential causes of complications prompted modification of our technique of PEG tube placement and management. Twenty-three patients received PEG for dysphagia/aspiration unrelated to neoplasia, 11 received PEG with staging endoscopy, 11 received PEG after treatment for head and neck neoplasm, and 26 received PEG at the time of primary resection. Major complications included retained PEG hub and delayed colon abscess ultimately resulting in death. Minor complications included skin abscesses, cellulitis, and early and late vasovagal response with PEG tube removal. An airway emergency, on attempted oral airway intubation, resulted in an aborted PEG attempt and constituted another complication outside the 4 groups stated above. The major complication was not found within a literature review. We have modified our management for avoidance of this complication. We believe the causes of the minor complications have been identified, and with additional modifications in our technique, we have not had any similar complications recently. The recommended techniques are discussed in detail.

Colon, Sigmoid↗

A functional model of nerve repair. Reanastomosis vs entubulation repair.

OBJECTIVE: To compare reanastomosis and entubulation repair after transection of the peroneal nerve in rats with the use of 2 functional models: gait analysis and a tension transduction device. DESIGN: Prospective, randomized, blinded control study. Each animal served as its own control. The injured site was alternated between the left and the right. Gait analysis was evaluated in a blind fashion at postoperative weeks 1, 3, 7, and 13, and the tension transduction device at week 13, for injured and uninjured legs. SUBJECTS: Animals underwent unilateral peroneal nerve transection injury, half with entubulation and half with end-to-end reanastomosis repair. INTERVENTION: Nerve transection was performed 6 mm proximal to the anterior tibialis muscle insertion followed by reanastomosis (epineurial suture placement to align the sectioned nerve ends) or entubulation (placement of nerve ends into a 4-mm-long sterile Silastic tube secured with 2 epineurial sutures). RESULTS: Gait analysis demonstrated a poorer ankle angle in all injured legs compared with uninjured legs at each postoperative period. Ankle angles for reanastomosis were statistically better than those for entubulation at weeks 3, 7, and 13. The tension transduction device demonstrated poorer force in injured than uninjured animals at 13 weeks. Reanastomosis repair groups demonstrated no difference in force development compared with entubulation repair groups. CONCLUSIONS: Reanastomosis of the transected rat peroneal nerve demonstrated improved functional return by gait analysis when compared with entubulation-repaired nerves at postoperative weeks 3, 7, and 13. Force development of injured nerve groups measured by the tension transduction device was decreased compared with control, but no difference was detected between the 2 repair methods. Further studies are needed to evaluate the possible functional benefit of reanastomosis.

Anastomosis, Surgical↗

Effects of differently induced stretch loads on neuromuscular control in drop jump exercise.

The neuromuscular characteristics of the triceps surae and vastus lateralis muscles and interactions between the pre-activation of these muscles and the muscle output itself during ground contact were investigated during various types of stretch-shortening cycle muscle loading. The loading of the muscles was effected by using three different types of drop jump exercise. These jumps allowed separate modifications of the loading of the leg extensor muscles by changing the velocity of the centre of gravity (CG) or by changing directly the body mass, which was also affected by changing artificially the acceleration of the CG. It was found that the eccentric peak angular velocity of the ankle joint was related to the various precontact and eccentric parameters in all the different types of jumping exercise. The correlations were higher for the gastrocnemius muscle (P < 0.001) than for the soleus muscle (n.s., P < 0.01). In all the experimental conditions, the pre-activation of the measured muscles started well before the impact of contact with the ground. However, the duration of the pre-activation phase depended on the type of stretch exercise. The results would suggest a clear interaction between the pre-activation of the muscles and that part of the muscle output which is effected by the segmental stretch reflex system. The control mechanism of the pre-activation itself appears to be multiple in character. It seems reasonable to assume that the pre-activation is preprogrammed, but which can, however, be modified by proprioceptive, vestibular and visual inputs. Thus, the possibility of conscious modification of the expected muscle load must be considered.

Adult↗

Hyperbaric oxygen treatment after rat peroneal nerve transection and entubulation.

Rat peroneal nerves were transected and entubulated with a Silastic channel. The experimental group was treated with hyperbaric oxygen to evaluate changes in acute edema, functional recovery, and histology. Hyperbaric oxygen was administered with 100% O2 at 2.5 atmospheres absolute for 90 minutes twice a day for 1 week and then four times a day for 1 week. Acute edema changes based on nerve water weight and transfascicular area measurements were greater in injured than in uninjured nerves but demonstrated no differences between hyperbaric oxygen-treated and -untreated groups 2, 8 and 16 days after surgery. Functional evaluation with gait analysis demonstrated significant changes between injured and uninjured group 1, 3, 7, and 13 weeks after injury but no differences between hyperbaric oxygen-treated and -untreated groups. Thirteen weeks after the initial injury, elicited muscle force measurements demonstrated no significant improvement from hyperbaric oxygen treatment of injured nerves. Histologic evaluation of nerve area, myelinated axon number, myelinated axon area, myelin thickness, and blood vessel number and area revealed no significant differences between hyperbaric oxygen-treated and -untreated groups. Hyperbaric oxygen was not associated with improvement of nerve regeneration with any of the outcome variables in this model.

Animals↗

Brain-derived neurotrophic factor and peripheral nerve regeneration: a functional evaluation.

The potential benefit of brain-derived neurotrophic factor (BDNF) on motor-nerve regeneration after transection injury in 24 adult rats was evaluated after entubulation repair. Gait analysis for ankle angle and tension transduction device (TTD) strain-gauge measurements yielded functional evaluation of regeneration. The BDNF (15 mg/mL) or phosphate buffered saline (control) was injected into the silicone elastomer (Silastic) channel. Gait analysis performed 0, 2, 4, 6, 10, and 12 weeks after injury demonstrated a significant difference between uninjured and injured legs of 23 and 43 degrees, respectively (P<.001, analysis of variance). The TTD evaluation 13 weeks after injury demonstrated a significant decrease in force development of injured compared to uninjured legs, 148 and 58 g, respectively (P<.001). No functional benefits were demonstrated between BDNF-treated versus control-treated animals in either model for a single exposure to BDNF.

Animals↗

Percutaneous endoscopic gastrostomy. A useful tool for the otolaryngologist-head and neck surgeon.

OBJECTIVE: To review our series of percutaneous endoscopic gastrostomy (PEG) tube placement by the Division of Otolaryngology. DESIGN: Charts from a total of 29 patients were reviewed; 23 patients with head and neck cancer and six patients with chronic aspiration. INTERVENTIONS: Placement of a PEG tube and other associated procedures, including primary tumor resection, tracheostomy, and surgical endoscopy. MAIN OUTCOME MEASURES: The feasibility, morbidity, and mortality of PEGs performed at the time of the primary surgical procedure compared with those being performed with a minor procedure. RESULTS: In almost all cases, the PEG was performed in conjunction with another procedure requiring general anesthesia, thereby decreasing the total number of procedure days. Morbidity and mortality were absent for all patients in whom PEG was performed. CONCLUSIONS: When properly applied, PEG can be performed by the otolaryngologist-head and neck surgeon with minimal or no morbidity at the time of staging or definitive procedure. Thus, the PEG can be of great benefit in patients with head and neck cancer.

Aged↗

Peroneal motor nerve crush injury and hyperbaric oxygen effect.

The potential therapeutic effect of hyperbaric oxygen (HBO) after rat peroneal nerve crush was evaluated. Animals were given 100% oxygen at 2.5 atmosphere absolute for 90 minutes twice daily for 1 week and then once daily for 1 week. Edema increased in crushed nerves compared with control nerves, but no effect was associated with the administration of HBO. Gait analysis demonstrated injury at 1, 7, and 14 days after nerve crush, but no difference was found at 22 and 28 days after injury (analysis of variance: P < .001, 10 animals per group). Eight weeks after injury, nerve stimulation and muscle force measurements were 114 g for the injured group and 146 g for the control group (P < .001). There were no HBO-associated changes in gait parameter or nerve/muscle force measurements. This study demonstrated that rat peroneal nerve crush injury causes acute intraneural edema and temporary decrement of gait parameters. Elicited nerve stimulation demonstrated persistent loss of force 4 weeks after normalization of gait, but no HBO effect.

Acute Disease↗

Tension transduction device for functional evaluation of the rat peroneal nerve.

Current techniques for evaluating animal model nerve regeneration lack accurate, sensitive and reproducible methods to determine neuromuscular function. We have developed a tension transduction apparatus which measures the magnitude of ankle dorsiflexion produced by normal rats during bipolar stimulation of the surgically exposed peroneal nerve. Three groups of animals were used to evaluate the consistency and overall reliability of this apparatus. Within the first group of 4 animals, we determined variability in a single testing period of 8 successive stimulations. The mean normalized standard deviation of dorsiflexion tension produced was 2.9% of the mean. In the second group, comparison of right and left dorsiflexion tension in 8 animals showed a difference of less than 2% (right: 134.2 g; left: 131.6 g), demonstrating that one lower limb can be used as a control for the contralateral limb. In the third group, 12 animals were tested on two separate occasions, 2 months apart, tension production increased 10% (from 122 to 134 g) and corresponded with an average weight increase, per animal, of 100 g (30%) during the 2-month rest period. Despite the increase in weight and strength, the initial procedure had a negative effect upon the maximum tension produced compared to a previously unoperated leg (prior operation: 134 g; no prior operation: 144 g). The reproducibility of data obtainable with this new device allows for its incorporation in future studies, as well as the correlation of such functional data with other current methods of studying nerve regeneration.

Animals↗

Risk factors associated with prolonged intubation and laryngeal injury.

A prospective study evaluated potential risk factors associated with laryngeal injury after prolonged endotracheal tube intubation for longer than 3 days. Ninety-seven patients were evaluated after oral endotracheal tube intubation (mean, 9 days). This study updates a previously reported evaluation of 44 patients. The additional sample size has provided findings of unreported patient risk factors of laryngeal injury and confirmation of previous associations. The majority of the 97 patients had some type of laryngeal injury, ranging from mild mucosal erythema to ulceration, granuloma formation, or true vocal cord immobility. Patient examinations were continued until the larynx returned to normal or stabilized or the patient was lost to follow-up. Postextubation examinations in the survival group revealed the following. (1) Laryngeal erythema occurred in 94%, and ulceration occurred in 76% of the patients with resolution within 6 weeks. (2) Laryngeal granulomas occurred in 44% of the patients; the majority of the granulomas (57%) developed an average of 4 weeks after extubation. Associated risk factors included duration of endotracheal tube intubation (p < 0.05) and presence of nasogastric tube (p < 0.05). (3) Vocal cord immobility was observed in 16 patients (20%). Eight patients had true vocal cord immobility noted initially after extubation, and the remaining eight had true vocal cord immobility an average of 4 weeks after extubation. Initial and delayed true vocal cord immobility were associated with duration of intubation and size of endotracheal tube (p < 0.01). Delayed true vocal cord immobility developed only in patients with a size 8 endotracheal tube.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Effect of artificial airway on ear complications from hyperbaric oxygen.

Hyperbaric oxygen treatment is associated with an increased risk of barotrauma to the tympanic membrane and middle ear. An artificial airway may compromise normal eustachian tube function and equilibration of middle ear pressures. This retrospective study was designed to evaluate the risk of middle ear complications in 267 patients receiving hyperbaric oxygen (HBO) therapy and to compare those with and without artificial airways. Charts of all patients were reviewed for middle ear and tympanic membrane complications and myringotomy tube placement. Eighteen of the 267 patients had artificial airways. Seventeen (94%) of these 18 patients developed middle ear or tympanic membrane complications, and 11 (61%) required tympanostomy tubes for pain, hemotympanum, or serous otitis. In contrast, 114 (45.8%) of the 249 patients without airways developed ear complications, and 53 (21.3%) required tympanostomy tubes. These results suggest that patients with an artificial airway who are receiving HBO therapy are at greater risk for developing tympanic membrane and middle ear complications than nonintubated patients. Similarly, patients with artificial airways receiving HBO frequently require placement of tympanostomy tubes.

Barotrauma↗

Effects of different simulated gravity conditions on neuromuscular control in drop jump exercises.

The neuromuscular characteristics of the triceps surae muscle were investigated during the various types of stretch-shortening cycle (SSC) muscle loading. The analysis concentrated on the preactivation and the contact phases of SSC. Muscle loading was changed unconventionally by artificially changing the condition of the gravity in drop jumps. This was accomplished by using a special lifting block system where the gravity could be modified to control loading and unloading effects of the triceps surae muscle. The normal gravity condition showed an advantage over the other gravity drop jump conditions for the measured parameters. The same tendency could be seen in the activation characteristics of the investigated muscles in the preactivation and eccentric phases. Further, the preactivation EMG was related to the eccentric peak angular velocity of the ankle joint. The correlation coefficients were 0.37 (p < 0.05) and 0.48 (p < 0.01) for the gastrocnemius and the soleus muscles, respectively. All the results emphasized considerable adaptation of the neuromuscular system to the normal gravity condition. However, the overall control of landing may also depend on the vestibular and visual inputs, which might modify even the earlier learned central programs.

Adaptation, Physiological↗