Data-intensive Undergraduate Research Project Informs to Advance Healthcare Analytics.

The overarching framework for incorporating informatics into the Wesley College (Wesley) undergraduate curriculum was to teach emerging information technologies that prepared undergraduates for complex high-demand work environments. Federal and State support helped implement Wesley's undergraduate Informatics Certificate and Minor programs. Both programs require project-based coursework in Applied Statistics, SAS Programming, and Geo-spatial Analysis (ArcGIS). In 2015, the State of Obesity listed the obesity ranges for all 50 US States to be between 21-36%. Yet, the Center for Disease Control and Prevention (CDC) mortality records show significantly lower obesity-related death-rates for states with very high obesity-rates. This study highlights the disparities in the reported obesity-related death-rates (specified by an ICD-10 E66 diagnosis code) and the obesity-rate percentages recorded for all 50 US States. Using CDC mortality-rate data, the available obesity-rate information, and ArcGIS, we created choropleth maps for all US States. Visual and statistical analysis shows considerable disparities in the obesity-related death-rate record-keeping amongst the 50 US States. For example, in 2015, Vermont with the sixth lowest obesity-rate had the highest reported obesity-related death-rate. In contrast, Alabama had the fifth highest adult obesity-rate in the nation, yet, it had a very low age-adjusted mortality-rate. Such disparities make comparative analysis difficult.

Formulation, pharmacokinetics and biodistribution of Ofloxacin-loaded albumin microparticles and nanoparticles.

Albumin microparticles containing Ofloxacin (Fluoroquinolone derivative commonly used in hospitals) were formulated by the spray dry method. By decreasing the pump feed rate or the total polymer concentration, a mixture of albumin/hypromellose acetate succinate (HPMCAS) microparticles and nanoparticles (MP/NP), containing Ofloxacin, were formulated. MP/NP were characterized, in vitro (particle size, zeta potential, and encapsulation efficiency). A comparison of the pharmacokinetics and biodistribution of aqueous Ofloxacin and Ofloxacin-loaded MP/NP, in Balb/c mice, revealed that peak concentrations were reduced in the serum, liver, spleen and brain, and a more sustained release was observed in serum and all of the organs tested for Ofloxacin MP/NP, compared to aqueous Ofloxacin. The MP/NP formulation allowed extended release by 24 h in the liver and more than 48 h in the brain. In serum, the elimination rate of Ofloxacin MP/NP is slower, the half life is longer, area under the plasma concentration time curve is decreased and volume of distribution is increased.

Development of sulforaphane-encapsulated microspheres for cancer epigenetic therapy.

Even though conventional chemotherapeutic management of cancer has reduced morbidity and mortality to a great extent, virtually all chemotherapeutic agents cause damage to healthy cells, necessitating exploration of novel anticancer agents that exert their effects through an alternate mode of action. Objectives of our research were twofold. First, we explored the promising potential of histone deacetylase inhibitor sulforaphane for epigenetic therapy for cancer as this therapeutic approach aims to reverse aberrant epigenetic modifications that affect gene expression. In vitro cell culture studies performed using B16 and S91 melanoma cells showed that sulforaphane inhibited growth and proliferation of cancer cells by downregulating deacetylation enzymes. The second part of our research investigated polymeric drug delivery systems to increase therapeutic efficacy and to minimize potential side effects of R,S-sulforaphane. Albumin microspheres encapsulating sulforaphane were developed by spray drying. Microspheres were characterized for their morphology, size and zeta potential. Cell culture studies using melanoma cells and in vivo studies in melanoma tumor-bearing C57BL/6 mice demonstrated that albumin based polymeric delivery system was efficacious and has the potential to enhance the therapeutic effect and anticancer activity of sulforaphane.

In vitro evaluation and targeting of E. coli internalized by endothelial cells using albumin microspheres loaded with gentamicin.

The purpose of this study was to evaluate the use of microencapsulated form of gentamicin and the traditional solution form for its intracellular bactericidal effect. Bovine serumalbumin (BSA) microspheres loaded with gentamicin were prepared by using Mini Spray Dryer. Human microvascular endothelial cells (HMECs) were exposed to increasing concentrations of Escherichia coli leading to internalization of E. coli. The internalized bacteria were targeted using either the microencapsulated or the solution form of gentamicin. The treatment groups using gentamicin solution form and microsphere form showed almost 46% and 86% inhibition in the growth of the internalized bacteria, respectively.

In vitro and in vivo evaluation of albumin-encapsulated primaquine diphosphate prepared by nebulization into heated oil.

Nebulization of an aqueous mixture of primaquine diphosphate and albumin into heated vegetable oil produces spherical particles with an average size of 6 microm. The microparticles are relatively stabile in buffers of pH 7.2 and 4.5 and completely degrade when exposed to proteolytic enzymes such as trypsin. Pharmacokinetic evaluation of the albumin-encapsulated primaquine diphosphate shows significantly higher levels in mouse liver tissue relative to free drug 2-48 h post-IP administration. Higher AUC (2.8x), lower steady-state volume of distribution (10x) and slower half-life (2.5x) relative to an equivalent dose of free primaquine diphosphate suggest liver targeting and sustained release of the drug from the microparticles.

Prevention of lethality and suppression of proinflammatory cytokines in experimental septic shock by microencapsulated CNI-1493.

CNI-1493, a newly developed, water-soluble tetravalent guanylhydrazone, is a powerful inhibitor of tumor necrosis factor (TNF) and interleukin-1 (IL-1) synthesis. Microencapsulation of drugs into microcapsules that target macrophages has improved the effectiveness of both TNF and IL-1 neutralizing antibodies in experimental models of septic shock. It is the purpose of this study to determine if microencapsulation of CNI-1493 will improve cytokine inhibition. CNI-1493 was microencapsulated using albumin into 1 microm spheres. Comparable amounts of CNI-1493 in solution and in microencapsulated form were added to 1 ml aliquots of whole blood along with 100 ng of endotoxin. TNF and IL-1 were measured by ELISA. Microencapsulated CNI-1493 was also given to rats with endotoxic shock (15 mg/kg Escherichia coli endotoxin) and rats with peritonitis induced by peritoneally injecting 10(10) CFU E. coli. Equivalent amounts of encapsulated and solution CN I-493 were given intravenously. Endotoxin 15 mg/kg was also given to rats 6 and 24 h after a dose of encapsulated CNI-1493 to determine the duration of action of encapsulated drug. The microencapsulated CNI-1493 produced significantly greater inhibition of TNF and IL-1 at all doses in the whole blood model. There was significantly improved survival and cytokine inhibition in the endotoxic shock model as well as the peritonitis model in rats treated with microencapsulated CNI-1493. There was also 83% survival in rats given endotoxin 24 h after a dose of encapsulated CNI-1493. From these data, we conclude that CNI-1493 is a potent inhibitor of cytokine production and is greatly potentiated by microencapsulation, which transports the drug to macrophages.

Sustained-release interleukin-12 microspheres in the treatment of cancer.

Interleukin-12 (IL-12) is a recently discovered cytokine with tremendous antitumor potential. It has been shown to boost the host immune response against experimental cancers in animal models. However, most studies have utilized IL-12 in the solution form, necessitating frequent dosing with higher doses, consequently leading to issues of toxicity. The only attempts at sustaining release have been in the production and use of genetically engineered cells that can secrete IL-12 constantly. These attempts are cost prohibitive and involve extensive labor. This study demonstrates the use of biodegradable albumin microspheres to sustain the release of IL-12. In vitro release of IL-12 from the microspheres was found to fit Higuchi's square-root-of-time model, suggesting diffusion-mediated release. About 46% of the theoretical IL-12 content was released slowly over a period of 24 hr. When administered intraperitoneally to C57BL/6 mice bearing subcutaneous melanomas, the microspheres significantly prolonged the survival when administered at half the weekly dose of the solution formulation. The microsphere dosage form also resulted in generally lower levels of liver and kidney function enzymes, suggesting lower toxicity.

Improved validated assay for the determination of mefloquine and its carboxy metabolite in plasma, serum and whole blood using solid-phase extraction and high-performance liquid chromatography.

An improved high-performance liquid chromatography method using a low silanol activity octadecylsilica column and a solid-phase extraction technique is validated for the simultaneous analysis of mefloquine and its carboxy metabolite in whole blood, plasma and serum. An octadecylsilica column with high silanol activity is compared to a column of low activity in terms of pH dependent variability of chromatographic retention times for mefloquine and its carboxy metabolite. The low silanol activity column showed a relatively large mobile phase pH range where retention times for both components are consistent. The solid-phase extraction procedure consists of a simple protein precipitation step followed by sample concentration and extraction using a C18 membrane disk. The inter- and intra-assay variability for a therapeutic concentration of mefloquine (1000 ng/ml) is less than 2% in whole blood, plasma and serum while carboxymefloquine (1000 ng/ml) is 2.3% or less. At concentrations as low as 100 ng/ml the inter-assay variability is 6.2% or less for both analytes. This method shows a robust analytical procedure for the simultaneous analysis of mefloquine and its carboxy metabolite where precise measurements are useful in pharmacokinetic studies and in estimating drug compliance.

Evaluation of microspheres containing cytokine neutralizing antibodies in endotoxemia.

Albumin microspheres are efficient carriers for delivering therapeutic agents to macrophages. In response to endotoxin, macrophages release tumor necrosis factor alpha (TNF alpha) and interleukin-1-beta (IL-1 beta). Blocking the effects of TNF alpha and IL-1 beta decreased lethality due to endotoxin-induced shock. In this study, we compared the efficacy of the microsphere form of TNF alpha and/or IL-1 beta neutralizing antibodies (NAs) with the free form of TNF alpha and/or IL-1 beta NA in preventing lethality due to endotoxemia and evaluated the duration of blockade by the microsphere form of TNF alpha and/or IL-1 beta NA on endotoxin-induced cytokine release. The results indicate that the microsphere form of TNF alpha and/or IL-1 beta NA protected 80% of the rats from lethal endotoxemia, while none of the rats that received the free from of TNF beta and/or IL-1 beta NA survived longer than 48 hr. The microsphere form of TNF alpha and/or IL-1 beta NA attenuated endotoxin-induced cytokine release more potently than the free form of TNF alpha and/or IL-1 beta NA in vivo. In vitro, the microsphere form of TNF alpha and/or IL-1 beta NA blocked endotoxin-induced cytokine release for at least 24 hr. Higher efficacy of the microsphere form of NA in reducing mortality and blocking cytokine release makes it more therapeutically advantageous than the free form of NA in the treatment of lethal endotoxemia.

Improved efficacy of a microencapsulated macrophage colony stimulating factor and methotrexate in melanoma.

This study examines the effects of a combination therapy of both methotrexate (MTX) and albumin microspheres containing recombinant human macrophage colony-stimulating factor (rhM-CSF) in melanoma tumors. Melanoma tumors were induced in C57BL/6 male mice with subcutaneous injection of B-16 tumor cells. Therapy started once the tumor size reached 0.5 cm in diameter. Mice were divided into several groups, and dosing was carried out daily until death. Group I received MTX solution (2 mg/kg or 15 mg/kg), group II received rhM-CSF solution (100 micrograms), group III received albumin rhM-CSF microspheres (100 micrograms), and groups V-XV received different combinations of both agents daily. The weight, tumor size, and survival time (in days) were recorded. From the results, the control (no rhM-CSF administered) group survived for 11.8 +/- 1.92 days, and the group that received MTX solution survived for 19.4 +/- 5.03 days. However, the group that received both the MTX solution (15 mg/kg) and albumin rhM-CSF microspheres (100 micrograms/kg) demonstrated a significant increase (p < .05) in the survival time (30.4 +/- 3.27 days). The concentrations of cytokines (tumor necrosis factor alpha [TNF-alpha] and interleukin-1 beta [IL-1 beta]) in the different treatment groups were monitored to determine the effect of rhM-CSF on the immune system. The TNF-alpha concentration was significantly higher in the group that received the combination therapy (204 +/- 54.6 pg/ml) versus the control group (31.5 +/- 7.02 pg/ml). The IL-1 beta concentration was significantly higher (p < .05) in the rhM-CSF microsphere (100 micrograms/kg) treated group (62 +/- 17.2 pg/ml) versus the rhM-CSF solution (29.1 +/- 8.7 pg/cc).

Macrophage depletion by albumin microencapsulated clodronate: attenuation of cytokine release in macrophage-dependent glomerulonephritis.

A macrophage plays an important role in mediating the inflammatory response. Cytokines released by activated macrophages contribute to inflammation in glomerulonephritis (GN). Clodronate, a biphosphonate, causes macrophage depletion when administered in an encapsulated form in liposomes. We used albumin as the polymer matrix to microencapsulate clodronate to the microspheres (MS) in the 1-micron size range. The purpose of this study was to (a) determine macrophage depletion by clodronate MS, (b) determine the effect of clodronate MS on endotoxin-induced cytokine release in vitro, and (c) assess the effect of clodronate MS on macrophage infiltration in experimental antiglomerular basement membrane nephritis. Macrophage depletion by clodronate MS was assessed by staining for the EDI marker. The results indicate greater than 95% depletion of macrophages from the spleen, liver, kidney, and blood. In the whole blood model, clodronate MS attenuated endotoxin-induced tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) release, and the attenuation by the microencapsulated form of clodronate was also more effective than the free (solution) form of clodronate. Macrophage infiltration into the glomerulus in experimental GN was also blocked very effectively by pretreatment with clodronate MS. In conclusion, macrophage depletion by clodronate MS may be beneficial in reducing cytokine release and renal damage in GN.

Effect of ciprofloxacin on the pharmacokinetics of multiple-dose phenytoin serum concentrations.

We performed this study to determine if an interaction exists during the co-administration of ciprofloxacin with phenytoin. Seven healthy volunteers received oral phenytoin, 200 mg/day, as a single dose for 10 days. On day 9, phenytoin blood sampling was performed at times 0, 1, 2, 4, 6, 8, 10, 12, and 24 h. On day 10, oral ciprofloxacin, 500 mg, b.i.d. was initiated. On day 14, blood samples were collected as previously described. Pharmacokinetic analysis was performed to determine if there were differences between the area under the concentration time curve (AUC), maximum serum concentration, Cmax, and time of maximum serum concentration, Tmax, of phenytoin before and during co-administration of ciprofloxacin. Four subjects completed the study. Results of the analysis showed no significant differences between AUC, Cmax, and Tmax of phenytoin before and during ciprofloxacin administration. However, one subject showed marked reductions in both AUC and Cmax. Similar reductions in plasma concentrations have also been reported, resulting in breakthrough seizures. In conclusion, ciprofloxacin was not shown to increase phenytoin plasma concentrations or AUC in healthy volunteers. The potential for decreasing plasma phenytoin concentrations may exist and warrants close monitoring of levels when these two agents are given simultaneously.

Increased carbamazepine plasma concentrations after fluoxetine coadministration.

The interaction between fluoxetine and carbamazepine was investigated in six normal, healthy male volunteers (aged 23 to 40 years). Subjects were given carbamazepine, 400 mg every morning, for 3 weeks. Venous carbamazepine blood samples were obtained at baseline and 1, 2, 4, 6, 8, 10, 12, and 24 hours after the morning dose. Fluoxetine, 20 mg every morning, was then coadministered with carbamazepine for 7 days. Venous carbamazepine blood samples were again obtained as described. Carbamazepine and carbamazepine-10,11-epoxide (CBZE) were assayed by HPLC. Addition of fluoxetine resulted in a significant increase in the area under the concentration-time curve of carbamazepine (105.93 +/- 18.05 micrograms/ml.hr versus 134.97 +/- 12.15 micrograms/ml.hr; t = 3.284; df = 5; p = 0.022) and CBZE (11.6 +/- 1.93 micrograms/ml.hr versus 15.2 +/- 2.4 micrograms/ml.hr; t = 2.805; df = 5; p = 0.038). Both oral and intrinsic clearance of carbamazepine was decreased significantly on fluoxetine addition (3.87 +/- 0.68 L/hr versus 2.98 +/- 0.26 L/hr; t = 3.025; df = 5; p = 0.029 and 17.90 +/- 4.9 L/hr versus 11.92 +/- 1.4 L/hr; t = 3.037; df = 5; p = 0.029, respectively). No significant changes were determined for fraction of absorbed dose, volume of distribution, absorption rate constant, and elimination rate constant. These findings suggest that fluoxetine can inhibit the metabolism of carbamazepine. Careful monitoring of patients is recommended when these two drugs are coadministered.

Inhibitory effect of cyclosporine A on the activity of oral hypoglycemic agents in rats.

Diabetic patients often have a kidney or pancreas allograft requiring cyclosporine A (CsA) to prevent transplant rejection. These patients usually take p.o. hypoglycemic agents to control their diabetes. Because of the reported adverse effects of CsA on glucose metabolism as well as its potential use in Type II diabetics, we were interested in evaluating the in vivo effect of CsA on the activity of p.o. hypoglycemic agents. We also wanted to determine if tolbutamide produced any adverse effects on the pharmacokinetics of CsA. Male, Holtzman rats were administered CsA (p.o.) followed 1 hr later with the hypoglycemic agent. Two hours later, blood samples were obtained to determine blood glucose levels. Animals treated with CsA alone produced a significant hyperglycemia. The hypoglycemic effects produced by tolbutamide and glyburide were inhibited in animals treated concomitantly with CsA. This inhibitory effect was not observed during the first 3 hr of CsA-treatment, could not be overcome by increasing the dose of the hypoglycemic agent and occurred using small doses. CsA did not, however, interfere with the activity of exogenous NPH insulin. Tolbutamide was found to have no effect on the acute pharmacokinetics of CsA. The distribution of CsA was similar to controls in all tissues studied except the liver in which CsA levels were less in tolbutamide-treated animals. These studies demonstrate that CsA interferes with the effects of p.o. hypoglycemic agents and, therefore, blood glucose levels should be monitored closely in Type II diabetic patients taking combinations of these drugs.

Pharmacokinetic analysis of cyclosporine in adjuvant arthritic rats.

Cyclosporine A (CsA) has been shown to be effective in patients with rheumatoid arthritis and to prevent the development and improve the symptoms of adjuvant-induced arthritis in rats. Since abnormal drug disposition has been reported in inflammatory conditions, we have evaluated the pharmacokinetics of CsA in this animal model of arthritis. We found a statistically significant decrease in the rate of disappearance of blood concentrations of CsA following iv administration to arthritic rats. The plasma half-life of CsA increased with a corresponding decrease in total body clearance. The volume of distribution remained unchanged. This abnormality in CsA kinetics was not observed in these animals until 10 days after adjuvant injection. The administration of CsA (15 mg/kg ip) twice daily to arthritic rats for 8 days produced a 39.8% and 49.5% inhibition of swelling in the right and left hindpaw, respectively. There was also a 63.5% decrease in the arthrogram score. Trough levels of CsA in arthritic animals were initially higher than in controls during this treatment but returned to control values after 8 days of dosing, suggesting reversal of abnormal disposition with improvement of the disease. The addition of indomethacin to the dosing regimen resulted in a significant increase in trough levels of CsA, indicating a drug interaction between these two compounds. Possible mechanisms responsible for these observations with CsA are discussed.