Biomedical subjects

K Banovac

Publications and source records attributed to K Banovac.

At least 19 recordsLinked to original sources

PubMed1997

Evaluation and management of heterotopic ossification in patients with spinal cord injury.

Sixty-three patients with paralysis secondary to spinal cord injury (SCI) were screened for heterotopic ossification (HO) by bone scintigraphy 27 +/- 14 (mean +/- SD) days after SCI. There were four female and 59 male patients, 36 had paraplegia and 27 tetraplegia. The age of patients was 28 +/- 9 years. Bone scintigraphy was obtained with a 3-phase test using 99m-technetium labeled diphosphonate, and the positive third phase was used as a criterion for diagnosis of HO. Bone scintigraphy was negative for HO in 27 patients (14 tetraplegic and 13 paraplegic) and positive in 36 patients (13 tetraplegic and 23 paraplegic). The patients with positive HO were treated with etidronate, first with an intravenous dose of 300 mg/day for 3 days, and then with an oral dose of 20 mg/kg/day for 6 months. The follow-up of patients consisted of clinical and radiographic evaluations every 2-4 months. The extent of HO was classified radiographically. In the treated group of patients who completed the entire course of etidronate therapy one patient developed HO, the remaining 28 (97%) patients had no radiographic evidence of HO during the follow-up of 10.6 +/- 4.5 months after initiation of therapy. Our data indicate that: (a) early HO can be detected in the asymptomatic patients using bone scintigraphy on the average of 4 weeks (27 +/- 14 days) after SCI and (b) the therapy with etidronate might be effective in the prevention of HO in majority of patients when the treatment is initiated in an early stage of heterotopic bone formation.

Adolescent↗

PubMed1997

Treatment of heterotopic ossification after spinal cord injury.

A new protocol in management of heterotopic ossification (HO) was evaluated in 46 patients after spinal cord injury (SCI). A group of 24 paraplegic and 22 tetraplegic patients was involved in a prospective study. Diagnosis of HO was made by bone scintigraphy and radiographic evaluation. Patients were divided into two groups. Group I was made up of 33 patients with positive bone scintigraphy and negative evidence of HO and Group II was made up of 13 patients with positive bone scintigraphy and positive radiographic evidence of HO. Etidronate was started intravenously (300 mg/day) for three days followed by oral therapy for six months (20 mg/kg/day). Follow-up of patients was 15.7 +/- 8 months after SCI. In Group I, etidronate therapy prevented the development of HO in 79 percent of patients; in 21 percent of patients, a low degree of tissue ossification was found which was not clinically significant. In Group II, there was an inhibitory effect of etidronate on progression of soft tissue ossification in six patients. The remaining seven patients did not respond to therapy and showed an increased growth of HO. Our data indicate that etidronate may prevent HO in the majority of patients when administered at an early stage of HO development and in higher doses than are routinely recommended.

Administration, Oral↗

PubMed1995

Interaction of cultured rat osteoblasts with 8-methoxypsoralen during irradiation with long-wave ultraviolet light.

Rat osteoblasts in monolayer cell cultures have been irradiated with long-wave ultraviolet light (UVA) in the presence and without 8-methoxypsoralen (8-MOP). In the absence of 8-MOP, the exposures to UVA (3 x 10(-3)W.cm-2) for up to 30 min have not affected cellular viability, the rate of 14C-acetate incorporation, and alkaline phosphatase (AP) activity. However, it depressed 3H-TdR incorporation rates by osteoblasts. In the presence of 15 to 100ng of 8-MOP/ml, even 5-min irradiation of osteoblasts was sufficient to reduce DNA synthesis. Much higher (0.5 to 1.0 micrograms/ml) 8-MOP concentrations were required to depress lipid synthesis, AP activity, and the viability of irradiated cells. These results suggest that in osteoblasts the machinery of DNA synthesis is especially labile to photosensitization with 8-MOP and UVA, whereas UVA light by itself exerts a less potent inhibitory effect.

Acetates↗

PubMed1995

Increased muscle strength in paralyzed patients after spinal cord injury: effect of beta-2 adrenergic agonist.

The administration of beta-2 adrenergic agonists in experimental animals result in an increased strength of skeletal muscle. In this study, we evaluated whether a beta-2 adrenergic agonist, metaproterenol, had an effect on muscle size and strength in a group of patients with muscular atrophy following spinal cord injury. Ten male subjects were randomly divided into 2 groups and agreed to participate in a prospective, double-blind, placebo-controlled, and crossover study. Metaproterenol (80 mg/day), or placebo, was administered orally for a period of 4 weeks. Muscle strength was measured by a force transducer interfaced with a microcomputer. Muscle size was calculated and expressed as a cross-sectional area of upper arm and forearm using a formula. Metaproterenol induced a significant increase of muscle strength in both groups of subjects, compared with placebo (p < .001). Similarly, there was an increase in a muscle size in the forearm following the administration of metaproterenol. Our data indicate that beta-2 adrenergic agonists may improve both muscle strength and size in patients with muscular atrophy following spinal cord paralysis.

Adult↗

PubMed1995

Effect of nonsteroidal antiinflammatory drugs on fracture healing: a laboratory study in rats.

We studied the effects of two nonsteroidal antiinflammatory drugs (NSAIDs) on fracture healing in rats: ibuprofen (30 mg/kg/day) and indomethacin (1 mg/kg/day). Femoral fractures were induced via a three-point bending technique. NSAIDs were administered orally for 4 or 12 weeks. Control animals received no medication. In each group a minimum of six animals were killed at the following intervals: 2, 4, 6, 8, 10, and 12 weeks postfracture. Fracture healing was determined by mechanical testing and histologic evaluation. The bending strength of each fractured femur was expressed as a percentage of the strength of the intact, contralateral femur. Histologic evaluation was performed on serial longitudinal sections stained with hematoxylin and eosin using a qualitative score of maturity of the callus. Ibuprofen and indomethacin both retarded fracture healing, with significant differences in "mechanical healing" found between the control and experimental groups after 10 weeks of drug administration. Both drugs also induced qualitative histologic changes manifested by delayed maturation of callus, which was noticeable earlier than the difference found by mechanical testing of bone. Our data suggest that NSAIDs have an inhibitory effect on fracture repair that is reversible after cessation of indomethacin but not ibuprofen.

Animals↗

PubMed1995

Fracture healing and mast cells.

We analyzed the morphology and localization of mast cells during the course of fracture repair in control rats and in animals with delayed healing of fractures induced by nonsteroidal antiinflammatory drugs (NSAIDs). In the first 2 weeks of fracture healing in control animals, mast cells were found either in the vicinity of blood vessels or in the vascularized tissue proliferating into the cartilaginous portion of subperiosteal callus. In the later stages (6-8 weeks), mast cells were seen in loose connective tissue in bone marrow surrounded with translucent ground substance. At this stage of healing, a hyperplasia of mast cells and cell degranulation was often seen in close proximity to osteoclasts and areas of bone resorption. Treatment with NSAIDs delayed fracture healing and the appearance of mast cell hyperplasia in bone marrow for approximately 4 weeks, suggesting that mast cells have specifically defined temporal and regional distribution during the process of bone repair. Histochemical studies documented a significant amount of chymase in the mast cells in callus. This enzyme was purified from mast cells by chromatography and was able to digest in vitro proteins extracted from bone. Our data suggest that mast cells in fracture healing are involved in digestion of extracellular matrix in callus tissue that could facilitate (a) angiogenesis in the early stages of healing, and (b) the replacement of provisional tissue with newly formed bone in the later stages of fracture healing.

Animals↗

PubMed1994

Gentamicin penetration into normal rabbit nucleus pulposus.

STUDY DESIGN: Radioactively labeled gentamicin was administered to 24 rabbits to assess the concentration of antibiotic in the nucleus pulposus. OBJECTIVES: The purpose of the study was to investigate the pharmacokinetics of gentamicin penetration into normal rabbit nucleus pulposus. SUMMARY OF BACKGROUND DATA: Disc space infection is a complication of spinal surgery that can be prevented by prophylactic antibiotics. Gentamicin can be used in conjunction with other antibiotics as a prophylactic agent. One previous study demonstrated that a similar antibiotic, tobramycin, penetrates the disc, but no data have been reported on the pharmacokinetics of disc penetration. METHODS: Twenty-four rabbits were given an intravenous injection of gentamicin labeled with iodine 125. Four rabbits were killed at hourly intervals 1 to 6 hours after injection. Specimens of nucleus pulposus, blood, whole liver, and saline-perfused liver were obtained and prepared. The radioactivity in the specimens was measured. RESULTS: The gentamicin concentration in the nucleus pulposus peaked at 2 hours and remained at this level for the duration of the experiment. Twenty percent of the gentamicin recovered from the nucleus pulposus was tissue bound. CONCLUSIONS: Gentamicin concentration in the rabbit nucleus pulposus does not peak until 2 hours after an intravenous bolus of drug. If gentamicin penetrates human nucleus pulposus in a similar fashion, this study could have implications for the timing of administration of this agent for prophylaxis.

Animals↗

PubMed1994

Tissue distribution of antibiotics in the intervertebral disc.

STUDY DESIGN: This study analyzed the distribution of antibiotics within the intervertebral disc of rabbits. Specimens were tested with specific antibodies against antibiotics using an immunofluorescent technique. OBJECTIVES: The results were correlated to provide a rationale for perioperative prophylaxis of infection. SUMMARY OF BACKGROUND DATA: Several groups of investigators and the recent data from our laboratory showed quantitative changes in penetration of antibiotics into intervertebral disc. No previous study has assessed antibiotic distribution in anulus fibrosus and nucleus pulposus. METHODS: Discs were obtained from rabbits after intravenous injection of penicillin or gentamicin. Antibiotics were localized in tissue sections using specific antibodies with a immunofluorescent method. RESULTS: Penicillin (negatively charged) and gentamicin (positively charged) penetrated the neutrally charged anulus fibrosus, but penicillin had less ability than gentamicin to penetrate into the negatively charged nucleus pulposus. CONCLUSION: Our data suggest that penetration and distribution of antibiotics into avascular intervertebral disc is significantly influenced by the charge of antibiotics.

Animals↗

PubMed1994

Evaluation of serum osteoblast mitogenic activity in spinal cord and head injury patients with acute heterotopic ossification.

STUDY DESIGN: This was a blind, prospective study of the effect of sera from patients with spinal cord and head injuries on osteoblast proliferation. OBJECTIVES: The authors studied whether a humoral factor that stimulates the formation of heterotopic bone is released into the circulation after a neural injury. BACKGROUND DATA: Other authors have shown that a humoral osteoinductive factor may be released after head and spinal cord injuries. METHODS: Serum was obtained at certain times throughout the first 12 weeks post-injury and from control subjects. It was incubated with osteoblasts harvested from fetal rats, as well as with fibroblast controls. RESULTS: There was a significant rise in serum mitogenic activity after injury in both groups. When patients that developed heterotopic ossification were compared to other patients and controls, no significant differences were seen. CONCLUSIONS: This in vitro study fails to support a humoral mechanism for heterotopic ossification after spinal cord or brain injuries.

Adult↗

PubMed1994

Morphologic and metabolic changes in rat osteoblast cultures during the dark reaction with 8-methoxypsoralen.

The dark reaction of 8-methoxypsoralen (8-MOP) with cultured rat osteoblasts did not cause significant changes in cellular replication rates or in the synthesis of RNA and proteins. Microscopic examination, however, revealed that the dark reaction resulted in massive accumulation of perinuclear lipids and in the statistically significant enhancement of alkaline phosphatase activity. A sharp, and statistically significant, upsurge of lipid synthesis in osteoblasts preceded microscopically detectable accumulation of lipids and occurred only during the initial, but not during the subsequent stages of the dark reaction. These results suggest that in the course of the dark reaction the plasma membrane of osteoblasts is a target of psoralen.

Alkaline Phosphatase↗

PubMed1993

Penetration of glycopeptide antibiotics in nucleus pulposus.

The penetration of the glycopeptide antibiotics vancomycin and teicoplanin into the nucleus pulposus of rabbits was studied. Blood samples were obtained at 0.5, 1, 4, 8, and 24 hours after intravenous administration of 30 mg/kg vancomycin or 16 mg/kg teicoplanin. Concentrations of antibiotics were determined in tissue specimens and serum samples by fluorescence polarization immunoassays. Antimicrobial activity in the nucleus pulposus was determined with an agar diffusion method using a strain of Micrococcus luteus as the indicator organism. A peak concentration of vancomycin in the nucleus pulposus was attained 8 hours after drug administration. Teicoplanin reached its maximum level and plateaued 1 and 2 hours, respectively, after injection, and it remained unchanged for the rest of the study. This microbiologic analysis showed that the nucleus pulposus contained an antimicrobial level of glycopeptide antibiotics after administration. Because rabbit nucleus pulposus is similar anatomically to that of humans, these results may have implications regarding the timing and choice of antibiotic administration.

Animals↗

PubMed1993

Intravenous disodium etidronate therapy in spinal cord injury patients with heterotopic ossification.

The goal of the present study was to use intravenous etidronate in the acute phase of heterotopic ossification (HO) in an attempt to achieve a high initial drug concentration at the site of the active ectopic ossification. The study included 27 consecutive patients with an acute onset of HO after spinal cord injury (SCI). The three-phase bone scan was used to confirm clinical diagnosis of HO. Disodium etidronate (Didronel) 300 mg was administered intravenously daily for 3 to 5 days. In 20 patients there was a rapid (1-2 days) decrease of soft tissue swelling (p < 0.01) with no side effects associated with the intravenous administration. In seven patients there was minimal or no improvement of edema after intravenous etidronate. In these patients deep vein thrombosis was found in the affected limbs. The effect of high dose etidronate on HO was determined in the group of 13 patients with positive clinical and scintigraphic finding of an acute HO, but negative radiographic studies. After intravenous administration of etidronate for 3 days (300 mg/day) the drug was continued orally with 20 mg/kg/day for 6 months. A placebo was not used in this study. In eight patients there was no radiographic evidence of HO after therapy while two patients had minimal ossifications. In three patients therapy was interrupted and all developed HO in 1-2 months.

Administration, Oral↗

PubMed1993

Interaction of osteoblasts with extracellular matrix: effect of mast cell chymase.

We studied the effect of mast cell chymase on the interaction between osteoblasts and extracellular matrix. Chymase was purified from mast cell lysate by anion exchange chromatography. Osteoblasts were isolated from rat calvarias by collagenase digestion. Incubation of osteoblasts with mast cell lysate (40-170 micrograms/ml) or purified chymase (8-32 micrograms/ml) resulted in changes in cell-matrix interaction and cell morphology. Osteoblasts treated with chymase also showed a gradual detachment from the artificial substrata and from the biomatrix (collagen-digested rib fragment). A similar effect of mast cell chymase on the osteoblasts was found in vitro on endosteum of an intact parietal bone. Neutral protease inhibitors abolished the effect of both crude and purified enzyme preparations on the cell-matrix interaction. Mast cell chymase had no effect on osteoblast viability. The effect of enzyme on osteoblast proliferation was studied with lower concentrations of enzyme (0.2 micrograms/ml) in order to avoid cell detachment; there was no effect on either the metaphase index or on the number of cells after 5 days of incubation with chymase. Osteoblast attachment and cell spreading on different matrix proteins (fibronectin, vitronectin, extract of noncollagenous matrix proteins from rat bone) were significantly altered by their pretreatment with chymase. Matrix fibronectin of osteoblasts in culture as well as soluble vitronectin and fibronectin were digested by rat mast cell chymase. Our data suggest that mast cells through action of neutral protease chymase may alter molecules in extracellular matrix that are important in osteoblast adhesion, cell spreading, maintenance of cell morphology, and, most likely, cell function.

Animals↗

PubMed1992

Effect of mast cell chymase on the morphology of thyroid cells in vitro.

We studied the effect of mast cell chymase on the thyroid cells in culture. Rat serosal mast cells, similar functionally to connective tissue mast cells, were obtained after lavage of the peritoneal cavity and lyzed by freezing. The resulting lysate was used as crude enzyme preparation. Mast cell chymase was purified from the crude preparation by anion exchange chromatography. Crude and purified chymase incubated with thyroid cells induced cellular retraction, the appearance of long processes and gradual cell detachment from the substratum. The effect of the enzyme was not cytotoxic. The immunofluorescence studies of thyroid cells showed a decreased amount of polymerized actin and tubulin after incubation with chymase. Neutral protease inhibitor abolished the effect of crude and purified chymase on thyroid cell morphology. The above findings suggest that mast cell chymase may have a function in the control of cell morphology and cell-matrix interaction.

Actins↗

PubMed1992

The effect of mast cell chymase on extracellular matrix: studies in autoimmune thyroiditis and in cultured thyroid cells.

In the first part of the study we analyzed the morphology of mast cells in autoimmune thyroiditis of BB/W rats. In the early stage of thyroiditis mast cells showed exocytosis of granules into the interstitium; this was associated with disorganization of the extracellular matrix and the appearance of a translucent ground substance in stroma. Mast cells were not seen in the mononuclear infiltrates in the later stages of thyroiditis. In order to further study the effect of mast cells on the extracellular matrix, we evaluated the effect of mast cell lysate and purified chymase on the matrix of cultured thyroid cells. Mast cells were obtained from peritoneal cavity; mast cell chymase was purified by anion exchange chromatography. After exposure to chymase there was a reduction of pericellular fibronectin in cultured thyroid cells, while laminin in matrix remained unchanged. Similarly, as found by gel electrophoresis, soluble fibronectin and vitronectin were digested by chymase in the reaction mixture. Cell attachment on both fibronectin and vitronectin was significantly decreased upon exposure of matrix proteins to chymase. The effects of chymase were abolished by enzyme inhibitor phenylmethane sulfonyl fluoride. These data suggest that mast cells possess proteolytic enzymes capable of digesting different host proteins which may have a role in the thyroid cell interaction with the surrounding matrix.

Animals↗

PubMed1991

Autoregulation of cerebral blood flow in patients with orthostatic hypotension after spinal cord injury.

Two groups of patients who developed orthostatic hypotension (OH) after spinal cord injury (SCI) were studied. In the first group all patients (4 females and 6 males) were asymptomatic, whereas in the second group (1 female and 9 males) all had clinical manifestations of hypotension. All but 3 patients were tetraplegic, and these patients were paraplegic above the T6 level. For this study blood pressure (BP), heart rate and cerebral blood flow (CBF) velocity were measured simultaneously on a tilt table at 0, 30, 60, and 80 degrees. Cerebral blood flow in the middle cerebral artery was measured bilaterally utilising the transcranial Doppler technique. In asymptomatic patients the mean baseline (0 degrees) BP (110 +/- 16/70 +/- 77 mm Hg systolic/diastolic) was not significantly different from the BP (106 +/- 16/68 +/- 11 mm Hg) of symptomatic patients. The mean maximal change in BP during tilting in the asymptomatic group (-23 +/- 10/10 +/- 7 mm Hg) was also not significantly different when compared to the symptomatic group (-29 +/- 13/11 +/- 6 mm Hg). CBF in the symptomatic group during the hypotensive reaction at 80 degrees was 32.5 +/- 5 cm/sec, while at the same body position in the asymptomatic group it was 40.9 +/- 8 cm/sec (significant at the p less than 0.02). In addition, CBF decreased in the symptomatic group at 80 degrees to 55.5 +/- 9.6% of baseline, while in the asymptomatic group the fall was 69.3 +/- 7.2% (p less than 0.001). Our data suggest that autoregulation of CBF rather than systemic BP plays a dominant role in the adaptation to OH in patients with SCI.

Adult↗

PubMed1991

Decreased incidence of urinary tract infections in patients with spinal cord injury: effect of methenamine.

This study was designed to determine the effect of methenamine on the frequency of urinary tract infections (UTI) in hospitalized patients after spinal cord injury. The study included 56 patients with neurogenic bladder dysfunction treated with intermittent catheterization. A group of 34 patients was treated with methenamine, 1g twice daily; the other group of 22 patients was the control group receiving no antimicrobial therapy. On a weekly basis urine samples from all patients were obtained for urinalysis and culture. A total of 500 urine samples were analyzed. The patients treated with methenamine had 23.4% positive urine cultures which was significantly lower than 57.5% in the untreated control group (p less than 0.001). Our data suggest that methenamine therapy is an effective prevention of UTI in paralyzed patients with neurogenic bladder dysfunction during the rehabilitation in hospital.

Bacteriuria↗

PubMed1989

A study of hepatic metabolism of thyroxine in BB/W rats treated with L-thyroxine.

The effect of thyroxine (T4) on T4 conversion to triiodothyronine (T3) and reverse T3 (rT3) was studied in BB/W rats. A colony of 38 BB/W rats was obtained and half were treated with thyroxine (T4), 1 mg per liter of drinking water. At 106 days of age the following groups were identified: nondiabetic, no T4 treatment, 8 rats; nondiabetic, T4 treated, 8 rats; diabetic, no T4 treatment, 10 rats; diabetic, T4 treated, 7 rats. All animals with diabetes were treated with insulin. T4 conversion to T3 and rT3 was assessed in liver homogenates in 0.1 M Tris-HCl buffer, pH 7.4, with or without 5 mM dithiothreitol (DDT). Serum T4 and rT3 were significantly elevated in both T4-treated groups (P less than 0.001), while serum T3 was not affected in either. Basal T4 deiodination to T3 by the liver homogenate did not change on treatment with T4; the addition of DTT increased T3 production in the homogenate from T4 treated nondiabetic animals (P less than 0.05). In both nondiabetic and insulin-treated diabetic rats there was no effect of T4 on the rate of rT3 production. Since, in the rat, 30-40% of circulating T3 is a direct contribution of thyroid gland secretion, and that would be absent in our T4-suppressed animals, the normal serum T3 may reflect increased absolute peripheral T3 production from the greater concentration of circulating T4.

Animals↗