Biomedical subjects

A K Mitra

Publications and source records attributed to A K Mitra.

At least 19 recordsLinked to original sources

PubMed2006

A comparison of stent-induced stenosis in coronary and peripheral arteries.

BACKGROUND AND OBJECTIVES: Restenosis is a complication of interventional procedures such as angioplasty and stenting, often limiting the success of these procedures. Knowledge regarding the relative behaviour of different arteries after these procedures is limited, despite the extensive use of different vascular models. Although the results from studies using different vessels are analysed to predict the behaviour of coronary arteries and other vasculature, direct controlled comparisons between different arteries are necessary for a better understanding of the differential response to restenosis. METHODS: This study examines the response to stenting in coronary and internal iliac arteries as characterised by intimal hyperplasia and restenosis. In a swine model of in-stent stenosis, coronary arteries exhibited higher levels of intimal hyperplasia and per cent stenosis than internal iliac arteries. RESULTS: After normalisation for injury score, coronary arteries were found to undergo 47% more intimal hyperplasia (p<0.05), whereas per cent stenosis normalised for injury score tended to be higher (p = 0.01). Other measurements reflecting post-stenting intimal hyperplasia (maximal intimal thickness, medial area) did not exhibit significant differences between the artery groups. CONCLUSIONS: These results show that coronary vessels are more prone to develop significant intimal hyperplasia and subsequent restenosis than internal iliac vessels. A better insight into how different arteries and arterial components behave is important in understanding and developing newer and better therapeutic measures for restenosis.

Animals↗

PubMed2006

In stent restenosis: bane of the stent era.

The long term outcome of stent implantation is affected by a process called in stent restenosis (ISR). Multiple contributory factors have been identified, but clear understanding of the overall underlying mechanism remains an enigma. ISR progresses through several different phases and involves numerous cellular and molecular constituents. Platelets and macrophages play a central role via vascular smooth muscle cell migration and proliferation in the intima to produce neointimal hyperplasia, which is pathognomic of ISR. Increased extracellular matrix formation appears to form the bulk of the neointimal hyperplasia tissue. Emerging evidence of the role of inflammatory cytokines and suppressors of cytokine signalling make this an exciting and novel field of antirestenosis research. Activation of Akt pathway triggered by mechanical stretch may also be a contributory factor to ISR formation. Prevention of ISR appears to be a multipronged attack as no therapeutic "magic bullet" exists to block all the processes in one go.

Apoptosis↗

PubMed2005

Comparing 2-year outcomes of anterior cruciate ligament reconstruction using either patella-tendon or semitendinosus-tendon autografts: a non-randomised prospective study.

PURPOSE: To compare the results of anterior cruciate ligament (ACL) reconstructions using either a patella-tendon autograft or a semitendinosus-tendon autograft. METHODS: Based on surgeon experience and preference, 68 patients underwent ACL reconstruction using either a quadruple-strand semitendinosus autograft (n = 34) or a central one-third bone-patella tendon-bone autograft (n = 34). Each patient was assessed preoperatively and postoperatively at 3, 6, and 24 months using the International Knee Documentation Committee (IKDC) knee score, Biodex muscle strength and endurance testing, and the KT1000 instrumented arthrometer test of knee laxity to anterior translation. All assessments at the 2-year follow-up were performed by the same physician and physiotherapist. RESULTS: While ACL reconstruction improved knee stability and IKDC knee scores significantly, there was no statistically significant difference between semitendinosus- and patella-tendon autograft reconstructions in terms of long-term knee score or laxity to anterior translation. Semitendinosus graft reconstruction was associated with less donor-site morbidity and hamstring weakness. Meniscectomy was associated with poorer long-term knee scores. CONCLUSION: ACL reconstruction is associated with a significantly better IKDC knee score and laxity measurement at 2-year follow-up. However, we were unable to demonstrate a significantly better long-term outcome in knee score or laxity to anterior translation with either a patella-tendon autograft or a semitendinosus-tendon autograft.

Adolescent↗

PubMed2004

Arsenic concentrations in rice, vegetables, and fish in Bangladesh: a preliminary study.

Arsenic contaminating groundwater in Bangladesh is one of the largest environmental health hazards in the world. Because of the potential risk to human health through consumption of agricultural produce grown in fields irrigated with arsenic contaminated water, we have determined the level of contamination in 100 samples of crop, vegetables and fresh water fish collected from three different regions in Bangladesh. Arsenic concentrations were determined by hydride generation atomic absorption spectrophotometry. All 11 samples of water and 18 samples of soil exceeded the expected limits of arsenic. No samples of rice grain (Oryza sativa L.) had arsenic concentrations more than the recommended limit of 1.0 mg/kg. However, rice plants, especially the roots had a significantly higher concentration of arsenic (2.4 mg/kg) compared to stem (0.73 mg/kg) and rice grains (0.14 mg/kg). Arsenic contents of vegetables varied; those exceeding the food safety limits included Kachu sak (Colocasia antiquorum) (0.09-3.99 mg/kg, n=9), potatoes (Solanum tuberisum) (0.07-1.36 mg/kg, n=5), and Kalmi sak (Ipomoea reptoms) (0.1-1.53 mg/kg, n=6). Lata fish (Ophicephalus punctatus) did not contain unacceptable levels of arsenic. These results indicate that arsenic contaminates some food items in Bangladesh. Further studies with larger samples are needed to demonstrate the extent of arsenic contamination of food in Bangladesh.

Agriculture↗

PubMed2004

Assessment of blood-induced cartilage damage in rabbit knees using scanning electron microscopy.

PURPOSE: To investigate the harmful effects of a single episode of intra-articular bleeding on articular cartilage of rabbit knees using scanning electron microscopy. METHODS: Autologous blood was injected into the right knee joints of 18 New Zealand white rabbits. Surface and cellular damages were examined by the scanning electron microscope (n=9) and light microscope (n=34), respectively. The injected right knees were then compared with the corresponding control left knees at one, 3, and 6 weeks after the blood injection. RESULTS: The articular surface of the injected knees turned uniformly rough with multiple pits after one week. Maximal changes with elevations and depressions were observed at 3 weeks. These changes reversed at 6 weeks with the irregularities smoothing out. A similar pattern of transient cartilage damage was noted histologically. CONCLUSION: Both scanning electron microscopic and light microscopic findings suggest that a single episode of intra-articular bleeding leads to articular cartilage damage but this appears to be reversible. Our findings of transient damage to the articular cartilage suggest that there is no need for intra-articular evacuation and washout following an acute episode of haemarthrosis.

Animals↗

PubMed2004

Morbidity following anterior cruciate ligament reconstruction using hamstring autograft.

INTRODUCTION: As anterior cruciate ligament (ACL) reconstruction becomes more frequently performed, the expectation of earlier return to activities becomes higher; hence graft selection becomes more important. The use of hamstring tendon autograft is perceived to have less postoperative morbidities. We evaluate the early postoperative complications in this prospective study. MATERIALS AND METHODS: Seventy-six patients operated by the same surgeon were assessed preoperatively, and 3 and 6 months after surgery. The hamstring and quadriceps strength were measured with Biodex machine and the side-to-side laxity measured using KT-1000 instrument. The patients were also assessed for their subjective complaints using IKDC (International Knee Documentation Committee) Knee Scoring System. RESULTS: The strength and endurance index of the quadriceps and hamstrings recovered and improved by the sixth month after surgery. The recovery by the quadriceps muscle is more marked and by the sixth month, it has recovered to more than the preoperative level. As for laxity, the average side-to-side difference at 30 pounds was 2.75 mm by 6 months. Of these patients, the side-to-side difference was less than 5 mm in 66 patients (5 mm as taken to be acceptable for graft success). As for IKDC rating, at the sixth month, 71 patients had normal or near normal knee by their own assessment. No patients had anterior knee pain at rest. Sensory deficits were only present in 3 patients with complaints of numbness over the distribution of the saphenous nerve or its infrapatellar branch. CONCLUSION: Most morbidities are temporal in nature and do not significantly affect the patient's activities.

Adolescent↗

PubMed2002

Time-dependent loss of radioactivity counts associated with paracellular markers in the presence of cyclodextrin.

This communication reports an unexpected phenomenon observed during the counting of radiolabeled paracellular marker solutions in the presence of 2-hydroxypropyl-beta-cyclodextrin (HPbetaCD). The results revealed time-dependent loss of 14C-mannitol and 14C-polyethlene glycol 4000 radioactivity counts with increased percentages of HPbetaCD. However, 14C-diazepam, a transcellular marker, displayed a stable count. A hypothesis behind this phenomenon is being proposed, involving water transfer from aqueous droplets to the surfactant rich scintillation fluid. The remaining droplets, becoming more and more concentrated in cyclodextrin, entrap the hydrophilic markers and consequently exhibit an increasing quenching effect. This effect shows that careful monitoring of radiolabeled markers used in transport experiments is necessary, even with high quench resistant scintillation fluids, to prevent erroneous interpretation of the transport data.

2-Hydroxypropyl-beta-cyclodextrin↗

PubMed2002

Intestinal absorption and biodistribution of cosalane and its amino acid conjugates: novel anti-HIV agents.

Cosalane and its amino acid conjugates are potent inhibitors of HIV replication. The purpose of this study was to investigate: (1) the pharmacokinetic disposition of the diglycine (GC) and the diaspartic acid (ASPC) conjugates of cosalane in male Sprague-Dawley rats; (2) intestinal absorption of cosalane and its amino acid conjugates using in vitro (small intestinal segments), in situ (closed loop); and (3) biodistribution of GC and its absolute oral bioavailability in rat. Cosalane and its conjugates exhibited biexponential disposition with very long half-lives upon intravenous dosing. However, these compounds failed to permeate the small intestine unless sodium desoxycholate (5-20 mM) was used as an intestinal permeation enhancer. A rank order correlation in terms of permeation enhancement in a descending order is as follows: GC>Cosalane>ASPC. In situ studies revealed that although the bile salt enhanced the permeation of cosalane across the enterocyte, its hepatic uptake was extensive. However, 66% of the absorbed dose of GC escaped uptake by the reticuloendothelial system (RES) and its biodistribution studies showed that the uptake by the RES was significantly lower compared to the parent compound. GC had an absolute oral bioavailability of 5.10+/-1.51%. Therefore, GC appears to be a favorable candidate for further development.

Animals↗

PubMed2002

Caffeine ameliorates radiation-induced skin reactions in mice but does not influence tumour radiation response.

Intramuscular administration of caffeine at a dose of 80 mg kg(-1) body weight to the gastrocnemius muscles of Swiss mice 5 min prior to local irradiation (35 Gy) of the leg delayed the progression of radiation-induced skin reactions in such animals. While 90% epilation with reddening of the skin was noted in animals treated with radiation alone, animals pretreated with caffeine suffered only partial hair loss with slight reddening of the skin on the 16th and 20th days post-irradiation. Beyond the 28th day, damage scores in irradiated feet for both the groups were similar (score 3) and remained unchanged until the 32nd day and then decreased and disappeared completely in both treatment groups by the 40th day after irradiation. In addition, the effect of caffeine on the radiation response of a mouse fibrosarcoma was investigated. Results showed that intratumoral administration of caffeine at a dose of 80 mg kg(-1) body weight 5 min prior to local exposure of tumours to 10 Gy of 60Co gamma-rays did not influence the response of tumours to radiation. The present study thus showed that although caffeine ameliorated radiation-induced skin reactions in the mouse leg, it did not affect the tumour radiation response, indicating its potential application in cancer radiotherapy.

Animals↗

PubMed2002

The posterior cruciate ligament: an anthropometric study in Asians and evaluation of safe limits for bony tunnel creation during reconstruction.

INTRODUCTION: Quantitative data regarding the dimensions of the posterior cruciate ligament (PCL) and its insertions have not been studied in Asians, and concerns have been raised regarding the danger of vascular injury when the bony tunnels are created during arthroscopically-assisted reconstruction. MATERIALS AND METHODS: Ten male cadavers were used in this study, incorporating dissection and procedural arms. In the procedural arm, the path of the drill bit was tracked fluoroscopically as the tibial and femoral tunnels were created during simulated reconstructive surgery, and the effect of varying knee flexion angles was studied. Fluoroscopic images were analysed using specialised image processing software. RESULTS: The mean length of the PCL at full extension was 37.7 +/- 1.9 mm, and the mean mid-substance width was 13.7 +/- 1.7 mm. The mean sagittal distance between the exit point of the tibial tunnel and the anterior surface of the popliteal artery, across all knee flexion positions, was merely 6.0 mm (range, 2.8 to 10.2 mm). This distance tended to increase with increasing knee flexion, but this was not statistically significant. The mean distance between the exit point of the femoral tunnel and the femoral artery was 51.1 mm (range, 42.1 to 59.0 mm). CONCLUSIONS: The dimensions of the PCL and its insertions in Asians do not vary greatly from those reported for Western subjects. The distance from the exit point of the tibial tunnel to the popliteal artery is very small and, although this distance increases with increasing knee flexion, the improvement in the safety margin may neither be clinically nor statistically significant. The margin of safety for drilling of the femoral tunnel is much greater.

Anthropometry↗

PubMed2001

Enhanced pulmonary delivery of insulin by lung lavage fluid and phospholipids.

Pulmonary delivery appears to be the most promising non-parenteral route of insulin administration. In this work, we investigated the enhancement of insulin absorption in the presence of phospholipids and lung lavage fluid in vivo and in vitro. In-vitro experiments of insulin uptake by type II cells showed a significantly enhanced absorption in presence of lavage fluid, compared to various buffer preparations. The same trend was obtained with in-vivo studies of tracheal instillation of insulin. The incorporation of phospholipids as absorption enhancers in 1,2-dipalmitoyl phosphatidylcholine (DPPC) dispersion was compared to blank liposomes. A significantly higher blood glucose decrease was observed with a DPPC-insulin physical mixture compared to liposome, suggesting a possible effect of the phospholipid chain physical state on the insulin in-vivo absorption.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed2001

Visualization of a water-selective pore by electron crystallography in vitreous ice.

The water-selective pathway through the aquaporin-1 membrane channel has been visualized by fitting an atomic model to a 3.7-A resolution three-dimensional density map. This map was determined by analyzing images and electron diffraction patterns of lipid-reconstituted two-dimensional crystals of aquaporin-1 preserved in vitrified buffer in the absence of any additive. The aqueous pathway is characterized by a size-selective pore that is approximately 4.0 +/- 0.5A in diameter, spans a length of approximately 18A, and bends by approximately 25 degrees as it traverses the bilayer. This narrow pore is connected by wide, funnel-shaped openings at the extracellular and cytoplasmic faces. The size-selective pore is outlined mostly by hydrophobic residues, resulting in a relatively inert pathway conducive to diffusion-limited water flow. The apex of the curved pore is close to the locations of the in-plane pseudo-2-fold symmetry axis that relates the N- and C-terminal halves and the conserved, functionally important N76 and N192 residues.

Amino Acid Sequence↗

PubMed2001

Inhibition of a model protease--pyroglutamate aminopeptidase by a natural oligosaccharide gum from Hakea gibbosa.

The purpose of this study was to investigate the effect of the oligosaccharide gum from Hakea gibbosa on the activity of a model protease enzyme pyroglutamate aminopeptidase (5-oxoprolyl peptidase; EC 3.4.19.3) and to elucidate the mechanism responsible for the decreased activity. Enzyme kinetic studies were conducted at 37 degrees C in 100 mM potassium phosphate buffer with 10 mM EDTA, 5% (v/v) glycerol, and 5 mM DTT (pH 8) for 15 min and were performed both in the presence and absence of the gum. Enzymatic activity was determined by a colorimetric assay using the specific substrate L-pyroglutamic acid beta-napthylamide. The enzyme kinetics was studied at various substrate and gum concentrations. The velocity of the reaction was determined by the amount of the product (beta-napthylamine) liberated at each substrate and gum concentration. The Ks and Vmax of the enzyme in the absence of the gum were 24.40+/-2.14 microM and 502.95+/-28.90 nmoles x min(-1) x mg protein(-1), respectively. As the concentration of the gum was gradually increased from 0.1 to 2%, the value of the Vmax decreased from 318.94+/-21.46 to 158.83+/-24.51 nmoles x min(-1) x mg protein(-1) while Ks increased from 17.42+/-4.6 to 63.03+/-1.89 microM. The mechanism for the inhibition of the enzyme by Hakea appeared to be a mixed-linear type (a type of non-competitive inhibition) as suggested from Hanes-Woolf, Dixon and Cornish-Bowden plots. The turnover number, kcat, calculated for the enzyme also decreased from 14.09+/-0.81 to 4.45+/-0.69 min(-1) as the concentration of the inhibitor was incrementally increased from 0 to 2% (w/v). The K(i) and alphaK(i) calculated from Dixon and Cornish-Bowden plots were found to be 0.31+/-0.11% (w/v) and 1.33+/-0.42% (w/v), respectively. The natural gum from Hakea gibbosa was effective in non-competitively inhibiting the enzyme pyroglutamate aminopeptidase. Thus, the natural gum may be a promising additive not only for its sustained-release and mucoadhesive properties as shown previously, but also for its ability to slow the enzymatic degradation of therapeutic polypeptides incorporated in dosage forms.

Hydrogen-Ion Concentration↗

PubMed2001

Nasal absorption of tyrosine-linked model compounds.

The objective of this research is to explore the amino acid carrier-mediated pathway for nasal uptake of tyrosine-linked model compounds. These molecules were synthesized by ether linkage between the aromatic hydroxyl group of L-tyrosine and the terminal hydroxyl group of model compounds. An enzymatically stable linkage was utilized to link the model compounds to L-tyrosine so that the loss of the compound from perfusate signals absorption only without complication from metabolism. The rat in situ nasal perfusion technique was utilized in this investigation. The analyte concentrations remaining in the nasal perfusates were quantitated by a reversed-phase high-performance liquid chromatography. All L-tyrosine-linked model compounds were absorbed across the nasal mucosa (15-50% absorbed), and the extent of nasal absorption was concentration dependent. The apparent first-order rate constants of intact conjugate loss from the nasal perfusate were calculated. The Michaelis-Menten kinetic model was applied to fit the perfusion data. The Michaelis constant, K(m), and maximum or saturation rate of compound loss from the perfusate, V(max), were calculated from Lineweaver-Burk plots [e.g., K(m) and V(max) are 0.57 mM and 0.00515 micromol/min, respectively, for one of the model compounds, O-(4-carboxyphenylmethyl)-L-tyrosine]. The absorption characteristics of all these model compounds exhibited saturable kinetics, whereas their parent compounds, possessing different octanol-water partition coefficients, showed either no absorption or transport by passive diffusion. Nasal uptake of L-tyrosine conjugates was inhibited by L-tyrosine. The results from this study indicate that nasal absorption of L-tyrosine-linked compounds is probably through an amino acid transport system. Exploitation of the amino acid transport pathway may be utilized to improve nasal absorption of poorly permeable drugs.

Administration, Intranasal↗

PubMed2001

Chemical stability, enzymatic hydrolysis, and nasal uptake of amino acid ester prodrugs of acyclovir.

The objective of this work was to improve nasal absorption of relatively impermeable small drug molecules via an amino acid prodrug approach. Acyclovir was selected as a model drug. L-Aspartate beta-ester, L-lysyl, and L-phenylalanyl esters of acyclovir were synthesized to investigate their effectiveness in enhancing nasal absorption of acyclovir. A stability study was conducted in phosphate buffer under various pH conditions at 25 and 37 degrees C. Enzymatic hydrolysis in rat nasal washings and plasma was conducted at 37 degrees C. A rat in situ nasal perfusion technique was utilized in this investigation to examine the rate and extent of nasal absorption of amino acid prodrugs. The remaining analyte concentrations in the nasal perfusate were quantitated by reversed-phase high-performance liquid chromatography. The results revealed that the L-lysyl and L-phenylalanyl esters were less stable than L-aspartate beta-ester. The stability of all three esters decreased with increasing pH and temperature. L-phenylalanyl ester is highly susceptible to plasma esterases, with an in vitro half-life 1.33 min. The rat in situ nasal perfusion study revealed that the extent of nasal absorption of acyclovir, L-lysyl and L-phenylalanyl esters was not significant (p < 1%). L-Aspartate beta-ester was absorbed to the extent of approximately 8% over 90 min of perfusion at an initial drug concentration of 100 microM. Nasal absorption of L-aspartate beta-ester of acyclovir was inhibited by L-asparagine but not by a dipeptide glycylsarcosine (Gly-Sar). The enhancement of acyclovir nasal absorption from the L-aspartate beta-ester prodrug suggests that nasal uptake of this prodrug probably involves an active transport system.

Acyclovir↗

PubMed2001

Binding of cosalane--a novel highly lipophilic anti-HIV agent--to albumin and glycoprotein.

Cosalane is a potent inhibitor of HIV replication with multiple sites of action. The purposes of this study were to (a) determine the extent and nature of cosalane binding to mucin, alpha(1)-acid glycoprotein (AAG), plasma, and human (HSA) and bovine serum (BSA) albumin, and (b) determine the primary site(s) of cosalane binding to HSA. Plasma protein binding of cosalane was studied by a gel filtration technique. Cosalane binding to HSA was also determined in the presence of salicylic acid. Competitive inhibition studies were conducted using warfarin, digitoxin, and diazepam to determine the primary HSA binding site(s) of cosalane. The drug was bound extensively to HSA and BSA and required 500-550 moles to saturate 1 mole of protein. Stoichiometries of cosalane binding to alpha(1)-acid glycoprotein (AAG) and mucin were between 30 and 50 mol/mol of either glycoprotein. The binding isotherm deviated from a rectangular hyperbola, suggesting self-association of the ligand. Salicylic acid decreased cosalane binding to HSA by one order of magnitude. Inhibition studies of cosalane to HSA revealed that the compound binds primarily to warfarin site with a K(i) of 1.24 +/- 0.24 nM. In summary, cosalane binds extensively to serum albumins and to a lesser extent to both AAG and mucin.

Animals↗

PubMed2001

Transport of acyclovir ester prodrugs through rabbit cornea and SIRC-rabbit corneal epithelial cell line.

The purpose of this study is to assess the permeability of acyclovir (ACV) prodrugs through the rabbit corneal cell line (SIRC) as well as the cornea, and characterize the SIRC cell line for transport and metabolism studies of ester prodrugs. Prodrug derivatization of an acycloguanosine antiviral agent, acyclovir, was employed to improve its permeability across the cornea. New Zealand albino rabbits were used as an animal model for corneal studies. The SIRC cell line grown on polyester membranes was used for transport of these prodrugs. SIRC cells grown on the membrane support for 10 days developed four to six layers of epithelial cells, and this is comparable to the normal rabbit corneal epithelial layer. Transport experiments were conducted across the rabbit cornea and confluent SIRC cells using side-by-side diffusion-cell apparatus. Enzymatic hydrolysis of these compounds was evaluated in SIRC cell lysates. Appropriate reversed phase HPLC method(s) were employed for quantitation of both the prodrug and ACV simultaneously. Corneal permeabilities of some of these prodrugs (Malonyl ACV and Acetyl ACV) were higher relative to ACV. The SIRC cell line permeability values of all the prodrugs were higher compared to that of the intact cornea. The total amount of ACV-prodrugs transported, i.e., unhydrolyzed prodrugs and regenerated ACV, across the SIRC cell line was more relative to ACV. Hydrolytic studies in the SIRC cell line homogenate demonstrated the bioreversion potential of the prodrugs and the presence of enzymes, particularly the cholinesterase in the SIRC cell line. It may be concluded that the SIRC cell line is leakier compared to the cornea. Keeping in mind the limitations, the SIRC cell line after further characterization may be used for transport and metabolism studies of ester prodrugs.

Acyclovir↗

PubMed2001

Ocular pharmacokinetics in rabbits using a novel dual probe microdialysis technique.

Ocular infections involve delicate internal structures of the eye that often require treatment with antimicrobial agents. A major constraint to the study of ocular drug absorption from systemic administration is the inaccessibility of the vitreous for continuous serial sampling. A novel dual probe microdialysis technique has been employed in our laboratory, which will enable the delineation of complete ocular pharmacokinetics of a drug. New Zealand albino rabbits weighing 2--2.5 kg were used. The animals were kept under anesthesia throughout the experiment. A concentric probe was implanted in the vitreous chamber about 3 mm below the corneal scleral limbus. Simultaneously a linear probe was implanted in the anterior chamber across the cornea. Intraocular pressure (IOP) was measured using Schiotz tonometer. The total protein concentrations in the aqueous and vitreous humors were determined using the Bio-Rad protein assay method. The aqueous and vitreous elimination kinetics of fluorescein were studied after intravitreal and systemic administrations over a period of 10 hr. Microdialysis technique was also compared to the conventional direct sampling technique by determining the intravitreal kinetics of fluorescein. Results suggest that IOP reverted to normal within 2 hr after the implantation of the probes. The increase in the vitreal total protein concentration was not significantly different from the baseline. The increase in the aqueous total protein concentration was less than five times the basal concentration throughout the experiment. The blood-aqueous and blood-retinal barrier integrity was delineated by determining the permeability index for fluorescein and were found to be 9.48 +/- 4.25% and 1.99 +/- 0.66% for the anterior and vitreous chamber, respectively. The rate constant of penetration of fluorescein into the anterior chamber was found to be 8.48 +/- 1.33 x 10(-2) min(-1), which was significantly higher than into the vitreous i.e. 4.34 +/- 2.82 x 10(-2) min(-1). The terminal elimination rate constant of fluorescein from the anterior chamber (1.48 +/- 0.79 x 10(-2) min(-1)) was found to be similar to that of the plasma terminal elimination rate constant (1.57 +/- 0.25 x 10(-2) min(-1)), but significantly higher than from the vitreous (3.0 +/- 0.7 x 10(-3) min(-1)). The terminal vitreal elimination rate constant of fluorescein after intravitreal administration was found to be similar by both microdialysis (3.98 +/- 0.6 x 10(-3) min(-1)) and direct sampling (4.38 +/- 1.4 x 10(-3) min(-1)) techniques. In case of direct sampling technique the area under the vitreous concentration-time curve was higher compared to that obtained by the microdialysis technique. There was no breakdown of the blood ocular barriers as shown by a very small change in the intraocular fluid protein concentrations. This was also confirmed by the fluorescein kinetics, which were in accordance with the previous studies. IOP data suggests that the intraocular fluid dynamics were not affected and the animals stabilized within 2 hr after the implantation of the probes. Fluorescein data suggests that the vitreous compartment is surrounded by a tighter barrier compared to the anterior chamber. This technique appears to be more sensitive, reproducible and requires only one animal for the determination of entire ocular pharmacokinetic profile.

Animals↗