Systems pharmacology and enhanced pharmacodynamic models for understanding antibody-based drug action and toxicity

Pharmacokinetic (PK) and pharmacodynamic (PD) models seek to describe the temporal pattern of drug exposures and their associated pharmacological effects produced at micro- and macro-scales of organization. Antibody-based drugs have been developed for a large variety of diseases, with effects exhibited through a comprehensive range of mechanisms of action. Mechanism-based PK/PD and systems pharmacology models can play a major role in elucidating and integrating complex antibody pharmacological properties, such as nonlinear disposition and dynamical intracellular signaling pathways triggered by ligation to their cognate targets. Such complexities can be addressed through the use of robust computational modeling techniques that have proven powerful tools for pragmatic characterization of experimental data and for theoretical exploration of antibody efficacy and adverse effects. The primary objectives of such multi-scale mathematical models are to generate and test competing hypotheses and to predict clinical outcomes. In this review, relevant systems pharmacology and enhanced PD (ePD) models that are used as predictive tools for antibody-based drug action are reported. Their common conceptual features are highlighted, along with approaches used for modeling preclinical and clinically available data. Key examples illustrate how systems pharmacology and ePD models codify the interplay among complex biology, drug concentrations, and pharmacological effects. New hybrid modeling concepts that bridge cutting-edge systems pharmacology models with established PK/ePD models will be needed to anticipate antibody effects on disease in subpopulations and individual patients.     

Seroprevalence of Borrelia burgdorferi infection among forestry workers and farmers in Duzce, north-western Turkey

Borrelia burgdorferi infection is the most frequent tick-transmitted disease worldwide. Our aim was to assess the seroprevalence of B. burgdorferi infection among forestry workers and farmers in Duzce, in the north-west region of Turkey. Blood samples from 349 forestry workers and farmers and 193 healthy blood donors were obtained to determine the presence of antibodies to B. burgdorferi. A two-step testing strategy was used; the sera were initially tested by ELISA and then by Western blot (WB) IgG. Demographic data regarding residence, age, gender, profession, tick bite history, contact with animals, and symptoms involving the skin, nervous system, and osteoarticular system were collected by questionnaire. All results were evaluated statistically using the chi2 test. The seroprevalence of B. burgdorferi was 10.9% (n=38) in forestry workers and farmers and 2.6% (n=5) in blood donors by ELISA, with a statistically significant difference (p<0.001). Seropositivity rates were related to age, gender, and common risk factors for the disease. IgG seropositivity was confirmed in four (1.1%) sera by WB. In this first seroepidemiological report from the northwest region of Turkey, tick bite exposure was found to be high, whereas B. burgdorferi infection was not common. Preventive measures against tick exposure and further studies to determine the distribution of Lyme disease in Turkey are proposed.     

Behavioral analyses of sugar processing in choice, feeding, and learning in larval Drosophila

Gustatory stimuli have at least 2 kinds of function: They can support immediate, reflexive responses (such as substrate choice and feeding) and they can drive internal reinforcement. We provide behavioral analyses of these functions with respect to sweet taste in larval Drosophila. The idea is to use the dose-effect characteristics as behavioral "fingerprints" to dissociate reflexive and reinforcing functions. For glucose and trehalose, we uncover relatively weak preference. In contrast, for fructose and sucrose, preference responses are strong and the effects on feeding pronounced. Specifically, larvae are attracted to, and feeding is stimulated most strongly for, intermediate concentrations of either sugar: Using very high concentrations (4 M) results in weakened preference and suppression of feeding. In contrast to such an optimum function regarding choice and feeding, an asymptotic dose-effect function is found for reinforcement learning: Learning scores reach asymptote at 2 M and remain stable for a 4-M concentration. A similar parametric discrepancy between the reflexive (choice and feeding) and reinforcing function is also seen for sodium chloride (Niewalda T, Singhal S, Fiala A, Saumweber T, Wegener S, Gerber B, in preparation). We discuss whether these discrepancies are based either on inhibition from high-osmolarity sensors upon specifically the reflexive pathways or whether different sensory pathways, with different effective dose-response characteristics, may have preferential access to drive either reflex responses or modulatory neurons mediating internal reinforcement, respectively.     

Impact of food concentration on diel vertical migration behaviour of <Emphasis Type="Italic">Daphnia pulex</Emphasis> under fish predation risk

Vertical migration of Daphnia represents the best-studied predator-avoidance behaviour known; yet the mechanisms underlying the choice to migrate require further investigation to understand the role of environmental context. To investigate the optimal habitat choice of Daphnia under fish predation pressure, first, we selected the individuals exhibiting strong migration behaviour. The animals collected from the hypolimnion during the daytime were significantly larger, being more conspicuous, and in turn performed stronger diel vertical migration (DVM) when exposed to fish cue. We called them strong migrants. Second, we provided the strong migrant D. pulex with food at high and intermediate (1 and 0.4 mg C l⁻¹, respectively) levels, which were well above the incipient limiting level and of high quality. They traded the benefits of staying in the warm water layer and moved down to the cold water in response to fish cue indicating fish predation. The availability of food allowed the animals to stay in the cold hypolimnion. However, at the low food level (0.1 mg C l⁻¹), which is an additional constraint on fitness, Daphnia moved away from the cold hypolimnion. Poor food condition resulted in strong migrant Daphnia to cease migration and remain in the upper warmer water layer. Although temperature is known to be a more important cost factor of DVM than food, our results clearly show that this is only true as long as food is available. It becomes clear that food availability is controlling the direction of vertical positioning when daphnids experience a dilemma between optimising temperature and food condition while being exposed to fish cue. Then they overlook the predation risk. Thus, the optimal habitat choice of Daphnia appears to be a function of several variables including temperature, food levels and fish predation. Handling editor: S. I. Dodson     

Daily Ambrosia Pollen Concentration in the Air of Ankara,Turkey (1990-1999)

The airborne ragweed pollen spectrum was investigated in the air of Ankara, Turkey for aperiod of ten years (1990-1999) using a Burkard seven-day volumetric recording trap. In our study period,long distance transported Ambrosia pollen has been registered. Daily pollen levels varied from low to highin Burge's system. In last three years, the pollen concentration of Ambrosia showed a clear increasingtendency. Our results prove that ragweed pollen may be an important threat for ragweed sensitive patientsin Ankara city in near future.     

Disruption of PTPRO causes childhood-onset nephrotic syndrome

Idiopathic nephrotic syndrome (INS) is a genetically heterogeneous group of disorders characterized by proteinuria, hypoalbuminemia, and edema. Because it typically results in end-stage kidney disease, the steroid-resistant subtype (SRNS) of INS is especially important when it occurs in children. The present study included 29 affected and 22 normal individuals from 17 SRNS families; genome-wide analysis was performed with Affymetrix 250K SNP arrays followed by homozygosity mapping. A large homozygous stretch on chromosomal region 12p12 was identified in one consanguineous family with two affected siblings. Direct sequencing of protein tyrosine phosphatase receptor type O (PTPRO; also known as glomerular epithelial protein-1 [GLEPP1]) showed homozygous c.2627+1G>T donor splice-site mutation. This mutation causes skipping of the evolutionarily conserved exon 16 (p.Glu854_Trp876del) at the RNA level. Immunohistochemistry with GLEPP1 antibody showed a similar staining pattern in the podocytes of the diseased and control kidney tissues. We used a highly polymorphic intragenic DNA marker-D12S1303-to search for homozygosity in 120 Turkish and 13 non-Turkish individuals in the PodoNet registry. This analysis yielded 17 candidate families, and a distinct homozygous c.2745+1G>A donor splice-site mutation in PTPRO was further identified via DNA sequencing in a second Turkish family. This mutation causes skipping of exon 19, and this introduces a premature stop codon at the very beginning of exon 20 (p.Asn888Lysfs*3) and causes degradation of mRNA via nonsense-mediated decay. Immunohistochemical analysis showed complete absence of immunoreactive PTPRO. Ultrastructural alterations, such as diffuse foot process fusion and extensive microvillus transformation of podocytes, were observed via electron microscopy in both families. The present study introduces mutations in PTPRO as another cause of autosomal-recessive nephrotic syndrome.     

The effect of synthesis media on the properties of substituted polyaniline/chitosan composites

Substituted polyaniline/chitosan (sPANI/Ch) composites were chemically synthesized in H₂SO₄ and CH₃COOH synthesis media. Structural and physical properties of the composites were characterized by using FTIR, SEM, TGA, UV–vis, XRD techniques, and conductivity measurements. The effect of synthesis media on morphology, thermal stability, conductivity, and crystalline properties was investigated. Chemical interactions between substituted polyanilines and chitosan were explained using FTIR spectra results. The different morphological surfaces were observed in SEM images of the composites. The size of the substituted polyaniline/chitosan (sPANI/Ch) composites was in nanoscale, and the composites synthesized in acetic acid media showed smaller structures than those of H₂SO₄ media and pure chitosan. It was interpreted from XRD results that the composites have amorphous structure and the PNEANI/Ch–CH₃COOH composite has the highest crystallinity.     

Effect of Ottoman Viper (Montivipera xanthina (Gray, 1849)) Venom on Various Cancer Cells and on Microorganisms

Cytotoxic and antimicrobial effects of Montivipera xanthina venom against LNCaP, MCF-7, HT-29, Saos-2, Hep3B, Vero cells and antimicrobial activity against selected bacterial and fungal species: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, E. coli O157H7, Enterococcus faecalis 29212, Enterococcus faecium DSM 13590, Staphylococcus epidermidis ATCC 12228, S. typhimirium CCM 5445, Proteus vulgaris ATCC 6957 and Candida albicans ATCC 10239 were studied for evaluating the potential medical benefit of this snake venom. Cytotoxicity of venom was determined using MTT assay. Snake venom cytotoxicity was expressed as the venom dose that killed 50 % of the cells (IC₅₀). The antimicrobial activity of venom was studied by minimal inhibitory concentration (MIC) and disc diffusion assay. MIC was determined using broth dilution method. The estimated IC₅₀ values of venom varied from 3.8 to 12.7 or from 1.9 to 7.2 μg/ml after treatment with crude venom for 24 or 48 h for LNCaP, MCF-7, HT-29 and Saos-2 cells. There was no observable cytotoxic effect on Hep3B and Vero cells. Venom exhibited the most potent activity against C. albicans (MIC, 7.8 μg/ml and minimal fungicidal concentration, 62.5 μg/ml) and S. aureus (MIC, 31.25 μg/ml). This study is the first report showing the potential of M. xanthina venom as an alternative therapeutic approach due to its cytotoxic and antimicrobial effects.     

A cradle-to-gate life cycle assessment of wood fibre-reinforced polylactic acid (PLA) and polylactic acid/thermoplastic starch (PLA/TPS) biocomposites

Purpose

Biopolymers are considered to be environmentally friendlier than petroleum-based polymers, but little is known about their environmental performance against petroleum-based products. This paper presents the results of a life cycle assessment (LCA) of two prototype biocomposite formulations produced by extrusion of wood fibre with either polylactic acid (PLA) or a blend of PLA and locally produced thermoplastic starch (TPS).

Methods

The study followed the LCA methodology outlined in the two standards set out by the International Organization for Standardization (ISO): ISO 14040 and ISO 14044 of 2006. A life cycle inventory (LCI) for the biocomposite formulations was developed, and a contribution analysis was performed to identify the significant inputs. Environmental performances of the two formulations were then compared with each other and polypropylene (PP), a petroleum-based polymer. The US Environmental Protection Agency’s impact assessment method, “TRACI: The Tool for the Reduction and Assessment of Chemical and Other Environmental Impacts”, was combined with Cumulative Energy Demand (a European method) in order to characterize the inventory flows. Environmental impact categories chosen for the analysis were the following: global warming, stratospheric ozone depletion, acidification of land and water, eutrophication, smog, human health (respiratory, carcinogenic, and non-carcinogenic) effects and ecotoxicity.

Results and discussion

We found that PLA is the significant input which contributes mostly to fossil fuel consumption, acidification and respiratory and smog effects. Impacts from PLA transport from the faraway source significantly added more burden to its contributions. TPS causes less environmental burden compared to PLA; the environmental performance of the biocomposite improved when a blend of PLA and TPS is used in formulating the biocomposite. The two formulations performed better than PP in all the environmental impact categories except eutrophication effects, which is important on a regional basis.

Conclusions

The following conclusions were drawn from this study:

• PLA is the environmentally significant input among the three raw materials.
• TPS causes less environmental burden than PLA. Environmental performance of the biocomposite improves in the life cycle energy consumption, fossil energy use, ozone depletion and non-carcinogenic impact categories when a blend of PLA and TPS is used.
• The biocomposite can outperform PP in all the impact categories except eutrophication effects if manufactured using hydroelectricity.

The biopolymer could be a potential alternative to PP as it could cause less of a burden to the environment on a cradle-to-gate basis. Environmental impacts at the complete life cycle levels should be looked into in order to fully understand its potential.     

The expression of HER-2/neu (c-erbB2), survivin and cycline D1 in serous ovarian neoplasms: their correlation with clinicopathological variables

Ovarian cancer is the most common cause of death among all gynecologic malignancies and a result of complex interaction of multiple oncogenes and tumor suppressor genes. The aim of this study was to evaluate expression of HER-2/neu (c-erbB2), survivin and cycline D1 biomarkers in serous ovarian neoplasms and their correlations with clinicopathological variables in serous ovarian cancers. We analyzed pathological specimens of 62 patients with benign (n = 25), borderline (n = 14) and malignant (n = 23) serous ovarian neoplasms. Immunohistochemical analysis was performed on formalin-fixed paraffin-embedded specimens. Significantly more immunoreactivity with HER-2/neu was detected in malignant tumors (100 %) compared to borderline (78.6 %) and benign tumors (48 %) (P < 0.01). Survivin expression was significantly higher in malignant tumors (91.3 %) than those found in borderline (71.4 %) and benign tumors (24 %) (P < 0.001). Similarly, higher cyclin D1 expression was observed in malignant tumors (95.6 %) compared to borderline (85.7 %) and benign tumors (48 %) (P < 0.001). Expression of all biomarkers analyzed significantly and gradually increased from benign to borderline and borderline to malignant serous tumors. In terms of clinicopathological variables, only tumor grade was associated with the expression of all biomarkers others exhibited different correlations in serous ovarian cancers. The expressions of HER-2/neu (c-erbB2), survivin and cycline D1 are positively correlated with the malignant potential of serous ovarian neoplasms.