Nanoparticle Based Combination Treatments for Targeting Multiple Hallmarks of Cancer.

Treatment of cancer remains one of the most challenging tasks facing the healthcare system. Cancer affects the lives of millions of people and is often fatal. Current treatment methods include surgery, chemotherapy, radiation therapies or some combinations of these. However, recurrence is a major problem. These treatments can be invasive with severe side effects. Inefficacies in treatments are a result of the complex and variable biology of cancerous cells. Malignant tumor cells and normal functioning cells share many of the same biological characteristics but the main difference is that in cancer cells there is in an overuse and over expression of these biological characteristics. These pertinent characteristics can be grouped into eight hallmarks, as illustrated by Hanahan and Weinberg. These characteristics include sustaining proliferative signaling, evading growth suppressors, resisting cell death, enabling replicative immortality, inducing angiogenesis, activating invasion and metastasis, reprogramming energy metabolism, and evading immune destruction. In order to provide a noninvasive, effective treatment, delivery methods must be explored in order to transport cytotoxic agents used for targeting the hallmarks of cancer in a safer and more effective fashion. The use of nanoparticles as drug delivery carriers provides an effective method in which multiple cytotoxic agents can be safely delivered to cancer tissue to simultaneously target multiple hallmarks. By targeting multiple hallmarks of cancer at once, the efficacy of cancer treatments could be improved drastically. This review explores the uses and efficacy of combination therapies using nanoparticles that can simultaneously target multiple hallmarks of cancer.

Significant lethality following liver resection in A20 heterozygous knockout mice uncovers a key role for A20 in liver regeneration.

Hepatic expression of A20, including in hepatocytes, increases in response to injury, inflammation and resection. This increase likely serves a hepatoprotective purpose. The characteristic unfettered liver inflammation and necrosis in A20 knockout mice established physiologic upregulation of A20 as integral to the anti-inflammatory and anti-apoptotic armamentarium of hepatocytes. However, the implication of physiologic upregulation of A20 in modulating hepatocytes' proliferative responses following liver resection remains controversial. To resolve the impact of A20 on hepatocyte proliferation and the liver's regenerative capacity, we examined whether decreased A20 expression, as in A20 heterozygous knockout mice, affects outcome following two-third partial hepatectomy. A20 heterozygous mice do not demonstrate a striking liver phenotype, indicating that their A20 expression levels are still sufficient to contain inflammation and cell death at baseline. However, usually benign partial hepatectomy provoked a staggering lethality (>40%) in these mice, uncovering an unsuspected phenotype. Heightened lethality in A20 heterozygous mice following partial hepatectomy resulted from impaired hepatocyte proliferation due to heightened levels of cyclin-dependent kinase inhibitor, p21, and deficient upregulation of cyclins D1, E and A, in the context of worsened liver steatosis. A20 heterozygous knockout minimally affected baseline liver transcriptome, mostly circadian rhythm genes. Nevertheless, this caused differential expression of >1000 genes post hepatectomy, hindering lipid metabolism, bile acid biosynthesis, insulin signaling and cell cycle, all critical cellular processes for liver regeneration. These results demonstrate that mere reduction of A20 levels causes worse outcome post hepatectomy than full knockout of bona fide liver pro-regenerative players such as IL-6, clearly ascertaining A20's primordial role in enabling liver regeneration. Clinical implications of these data are of utmost importance as they caution safety of extensive hepatectomy for donation or tumor in carriers of A20/TNFAIP3 single nucleotide polymorphisms alleles that decrease A20 expression or function, and prompt the development of A20-based liver pro-regenerative therapies.

The thermal biology of digestion in rubber boas (Charina bottae): physiology, behavior, and environmental constraints.

Coadaptation predicts a match between the thermal physiology and thermoregulatory behavior of reptiles. We tested this prediction by studying the digestive biology of rubber boas (Charina bottae). We measured the thermal dependence of gastric digestive rate and passage rate in rubber boas from 10 degrees C to 35 degrees C. We examined the effect of digestion on their thermal preference by measuring the temperatures of C. bottae in a thermal gradient before and after feeding. While the passage rates calculated from the body temperatures of digesting snakes were higher than the passage rates calculated from the body temperatures of nondigesting snakes, there was no difference in calculated digestive rates. These results indicate that the thermoregulatory behavior of C. bottae may be more tightly correlated with factors affecting passage rate than with digestive rate alone. Results of simulating the constraints of the thermal environment on the digestive biology of C. bottae showed that digestion would take more than twice as long in the spring as in the summer. In addition, during the summer, snakes thermoregulating as digesting snakes would pass food 12% faster than those thermoregulating as nondigesting snakes. These results demonstrate how interpretation of laboratory studies can be improved when combined with measurements of appropriate environmental conditions.

Disease management program for asthma: baseline assessment of resource use.

Resource use by asthma patients was assessed as a step in the development of a disease management program by a university teaching hospital, a health maintenance organization (HMO), and a national pharmacy benefit management company (PBM). Medication profiles and medical records were reviewed for all HMO patients who had a diagnosis of asthma and a pharmacy claim for an asthma-related drug in 1993; patients with chronic obstructive pulmonary disease were excluded. These 656 patients' use of health care resources (outpatient clinic visits, emergency room and urgent care visits, hospital admissions, and, the associated costs were determined, as were variances from the PBM's clinical guidelines for asthma. Members 0-4 years of age had the most outpatient and emergency/urgent care visits and the most hospital admissions. Forty-four of the patients received high-dose beta-agonist therapy, and 20 of these patients did not receive either an inhaled anti-inflammatory drug or a short course of corticosteroids. The 44 patients had more outpatient clinic and emergency room/urgent care visits, more admissions, and greater total health care costs than the other patients with asthma. The asthma patients' mean health care cost in 1993 was $203, compared with $110 for all enrollees in the HMO. For patients with high use of beta-agonists, the mean cost was nearly three times that for the other asthma patients. The baseline review of resource use identified aspects of medication use that were at variance with treatment guidelines for asthma patients.

Development of standards for providing prescription drugs by mail.

The development and implementation of practice standards for a mail-service pharmacy are described. Literature, licensing regulations, and existing national practice standards were reviewed. Selected mail-service pharmacies were surveyed on their practice standards. Patients who had received prescription drugs by mail from a university hospital pharmacy were asked their opinions of the service in a telephone survey. Based on the information gathered, a set of practice standards was drafted. The draft was considered by a panel of experts who reached a consensus on mail-service pharmacy practice standards. The standards were implemented at a university hospital's outpatient pharmacy. Standards of practice for mail-service pharmacies were developed and applied at a university hospital outpatient pharmacy. Other pharmacies may find the standards useful.

Evaluation of prescription charges in a health maintenance organization.

Differing patterns in drug use and charge variances among three pharmacy sites were determined for a university-based health maintenance organization (HMO). Computerized prescription claim data for the HMO inhouse pharmacy and those chain and independent pharmacies contracting with the HMO were analyzed on a personal computer using database management and statistical software. Chain and independent pharmacies were under contract to charge the average wholesale price plus a $2.00 fee per prescription. Drug-claim data were separated by pharmacy site (inhouse, chain, and independent), and the data for each site were sorted by national drug code number. High-charge and high-use drugs were identified for evaluation on a unit-charge basis. During the six-month study period, 22,393 prescriptions were filled, representing total charges of $347,063.79. The mean per-prescription charges submitted to the HMO by the chain, independent, and inhouse sites were $14.99, $16.08, and $16.41, respectively. The inhouse pharmacy had many charges in high dollar ranges. Of 10 high-charge drugs analyzed, isotretinoin 40 mg, flunisolide nasal solution, naproxen 250 mg, cyclosporine oral solution, and atenolol 50 mg were dispensed in significantly different proportions at each site. There were significant differences in unit prices of nine high-use drugs among pharmacy sites. This study indicates the importance of evaluating adverse selection among pharmacy sites and the value of auditing claims submitted.

Two forms of RPO41-dependent RNA polymerase. Regulation of the RNA polymerase by glucose repression may control yeast mitochondrial gene expression.

We have identified two chromatographically separable forms of mitochondrial RNA polymerase from Saccharomyces cerevisiae which utilize different DNA templates. One form is only active in a nonselective assay utilizing a poly[d(A-T)] template. The other form selectively initiates from a mitochondrial promoter consensus sequence. Both enzymes can be extracted from yeast mitochondria and all components are encoded by nuclear genes. The possibility that these two activities represent core and holoenzyme forms of the multicomponent mitochondrial RNA polymerase is supported by our observation that both enzymes are absent from a strain bearing a disrupted copy of the RPO41 gene (Greenleaf, A. L., Kelly, J. L., and Lehman, I. R. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 3391-3399). The two enzyme activities are differentially regulated by carbon source; the nonselective enzyme is repressed during growth on glucose relative to the selective enzyme. The 5-fold increase in RNA polymerase activity on a nonrepressing carbon source correlates with the increased level of transcript production from mitochondrial DNA. These results suggest that the mitochondrial RNA polymerase and, in consequence, mitochondrial transcription are regulated by carbon catabolite control.

Constancy of ribosomal RNA genes during aging of mouse heart cells and during serial passage of WI-38 cells.

The DNA content and ribosomal RNA gene copy number in heart of the inbred mouse strain C57BL/6 were determined at different ages. The DNA content of mouse heart remained constant, at about 150 micrograms DNA per heart, from 1 to 30 mth of age. The number of rRNA genes, as estimated by 28S rRNA . DNA hybridization, was not found to change significantly as a function of age. Likewise, the extent of rRNA hybridization to DNA from cultured human WI-38 cells at early and late passage levels was the same. These data support the notion that genomic rDNA sequences are not lost during in vivo and in vitro aging. However, the rDNA sequences are quite large and numerous small deletions or base pair substitutions would not have been detected in these studies.

Time and cost analysis of a computerized outpatient pharmacy.

The time and cost of implementing and operating a computerized outpatient pharmacy was compared with similar components of a traditional system. Time and motion observations were used to analyze the time required to complete each component of the dispensing process for outpatient prescriptions and clinic-administered medications. Capital equipment expenditures and operating costs were compared for both systems. The computerized dispensing process for outpatient prescriptions showed a significant decrease in total dispensing process time, attributed primarily to the clerical component. Total dispensing process times for clinic-administered medications were not significantly different for the two systems. As compared with the traditional system, the computerized system's implementation and operating costs were higher for both outpatient prescriptions and clinic-administered medications. The computerized system decreased the time needed to complete the dispensing process but at an increased cost when compared with a traditional dispensing system.

Familial spastic paraplegia: a clinical and electrodiagnostic evaluation.

Twenty members of a family with an autosomal dominant variety of a familial spastic paraplegia (FSP) underwent neurologic and electrodiagnostic examination. The degree of clinical involvement ranged from severe to none. This study investigated the presence of peroneal H reflex of Hoffmann along with other electrodiagnostic parameters in each family member and compared this with the presence of other neurologic findings. An H reflex in the peroneal nerve was found in all subjects with a definite diagnosis of FSP and in 4 of the 6 subjects with a diagnosis of probable FSP. It was not elicited in any subject with fewer than 3 neurologic signs or below the age of 25. The H reflex was found to have value as an indicator of upper motor neuron involvement when taken together with the neurologic findings, but its predictive value for individuals at risk to develop FSP has not been established.

Slowly progressive autosomal dominant spastic paraplegia with late onset, variable expression and reduced penetrance: a basis for diagnosis and counseling.

We have examined a pedigree in which familial spastic paraplegia (FSP) is segregating in four generations. The data show a high rate of transmission of the trait, late onset, reduced penetrance, variable age and symptom expressivity, and an autosomal dominant mode of transmission. A summary of our data together with the FSP data of other shows a 1:1 transmission from males and from females, and an overall 1:1 transmission ratio. The risks for the children of symptomatic and non-symptomatic parents are illustrated.

Some determinants of the reinforcing and punishing effects of timeout.

Some determinants of the reinforcing and punishing properties of timeout were investigated in two experiments. Experiment I began as an attempt to reduce the frequency of tantrums in a 6-yr-old autistic girl by using timeout. Unexpectedly, the result was a substantial increase in the frequency of tantrums. Using a reversal design, subsequent manipulations showed that the opportunity to engage in self-stimulatory behavior during the timeout period was largely responsible for the increase in tantrums. Experiment II was initiated following the failure of timeout to reduce the spitting and self-injurious behavior of a 16-yr-old retarded boy. Using a multiple-baseline design, the nature of the timein environment was shown to be an important determinant of the effects of timeout. When the timein environment was "enriched", timeout was effective as a punisher. A conception of timeout in terms of the relative reinforcing properties of timein and timeout and their clinical implications are discussed.