Rate Of Metabolic Activity Research Articles

Accumulation of amino-polyvinyl alcohol-coated superparamagnetic iron oxide nanoparticles in bone marrow: implications for local stromal cells.

First, it will be investigated if amino-polyvinyl alcohol-coated superparamagnetic iron oxide nanoparticles (A-PVA-SPIONs) are suitable for MRI contrast enhancement in bone marrow. Second, the impact of A-PVA-SPION exposure in vivo on the viability and key functions of local bone marrow stromal cells (BMSCs) will be investigated. Animals were systemically injected with A-PVA-SPIONs, followed by a 7-day survival time. Accumulation of A-PVA-SPIONs was confirmed by MRI, histology and inductively coupled plasma optical emission spectrometry. BMSCs were isolated from bone marrow for in vitro assessment of their viability and regenerative key functions. In this study, A-PVA-SPIONs were found to accumulate in bone marrow and increase the BMSCs' metabolic activity and migration rate. A-PVA-SPIONs appear suitable for contrast enhancement in bone marrow while our data suggest an influence on the BMSCs biology that necessitates future research.

Using a Vulnerability Theoretical Model to Assess the Malignant Hyperthermia Susceptible Population: Implications for Advanced Practice Emergency Nurses.

Malignant hyperthermia (MH) is a high-risk, low-occurrence medical emergency with symptoms that include a severe increased rate of metabolic activity and rigid skeletal muscles. Clinicians should be knowledgeable and prepared for an MH event because it can occur in areas outside the operating room and without anesthetic triggers. Patients who have this rare genetic condition may come to the emergency department (ED) presenting with symptoms of heat stroke. However, the incidence of suspected MH in the ED or other critical care areas is not easily quantifiable because clinicians may not report cases to a centralized registry. The purpose of this article is to describe the MH-susceptible vulnerable population, to apply a vulnerability theoretical model to assess patients and families, to identify strategies for health promotion to reduce vulnerability, and to discuss how advanced practice nurses who specialize in emergency care can help decrease the vulnerability of MH-susceptible patients and families. By using a vulnerability model to assess the MH-susceptible population, nurses can effectively sort out strategies to prevent poor patient outcomes related to MH and promote health for this high-risk population. Measuring accurate core temperatures, applying effective cooling methods, and administering dantrolene are key concepts in caring for a patient who is experiencing an MH event. Advanced practice emergency nurses can participate in reducing vulnerability for this population by applying the Emergency Nurses Association Clinical Nurse Specialist competencies to MH-related vulnerabilities. Enhancing preparedness, evaluating and coordinating education programs, advocating for report submissions to the North American Malignant Hyperthermia Registry, and assessing opportunities for community collaboration are among the strategies discussed for reducing vulnerability for the MH-susceptible population.

Closed circuitry operation influence on microbial electrofermentation: Proton/electron effluxes on electro-fuels productivity

A novel biocatalyzed electrofermentor (BEF) was designed which uncovers the intricate role of biocatalyst involved in cogeneration of electro-fuels (hydrogen and electricity). The specific role of external resistance (Rext, electrical load) on the performance of BEF was evaluated. Four BEFs were operated separately with different resistances (25, 50, 100 and 200Ω) at an organic load of 5g/L. Among the tested conditions, external resistance (R3) with 100Ω revealed maximum power and cumulative H2 production (148mW and 450mL, respectively). The competence of closed circuitry comparatively excelled because it facilitates congenial ambiance for the enriched EAB (electroactive bacteria) resulting high rate of metabolic activity that paves way for higher substrate degradation and electro-fuel productivity. Probing of electron kinetics was studied using voltammetric analyses wherein electron transfer by redox proteins was noticed. The designed BEF is found to be sustainable system for harnessing renewable energy through wastewater treatment.

Synergistic yield of dual energy forms through biocatalyzed electrofermentation of waste: Stoichiometric analysis of electron and carbon distribution

A novel BEF (biocatalyzed electrofermentor) is designed by hybridizing the functional properties of both MFC (microbial fuel cell) and acidogenic fermentation process (AFP). This prototype facilitates potential synergy between the electrogenic and acidogenic processes to recover biohydrogen and bioelectricity with simultaneous wastewater treatment. The BEF was operated in three circuitry modes of operation viz., OC (open circuit), SC (short circuit) and CC (closed circuit) and the performance was compared with a control (without electrode assembly) with DSW (designed synthetic wastewater) having an organic load of 5000 mgL−1. In comparison with other test conditions, CC mode with fixed external resistance (300 Ω) gave highest yields of power density (72 mW m−2) and biohydrogen production (343 mL). Besides, the BEF performance was sustained by the innate buffering capacity and the substrate-linked dehydrogenase enzyme activity. The CC mode comparatively excelled because it facilitates congenial ambiance for the enriched EAB (electroactive bacteria) resulting high rate of metabolic activity that paves way for higher substrate degradation and product conversion efficiency. The empirical analysis of electron and carbon distribution was in good agreement with the experimental results. The electron delivery kinetics studied using voltammetric technique confirmed electron transfer by the membrane bound redox mediators. The designed biocatalyzed electrofermentation unravels the scope to harness dual forms of energy along with waste remediation.

Metabolic decoupling in daily life in patients with panic disorder and agoraphobia

Various studies have assessed autonomic and respiratory underpinnings of panic attacks, yet the psychophysiological functioning of panic disorder (PD) patients has rarely been examined under naturalistic conditions at times when acute attacks were not reported. We hypothesized that emotional activation in daily life causes physiologically demonstrable deviations from efficient metabolic regulation in PD patients. Metabolic coupling was estimated as within-individual correlations between heart rate (HR) and indices of metabolic activity, i.e., physical activity (measured by 3-axial accelerometry, Acc), and minute ventilation (Vm, measured by calibrated inductive plethysmography, as proxy for oxygen consumption). A total of 565 daytime hours were recorded in 19 PD patients and 20 healthy controls (HC). Pairwise cross-correlations of minute-by-minute averages of these metabolic indices were calculated for each participant and then correlated with several indices of self-reported anxiety. Ambulatory HR was elevated in PD (p = .05, d = 0.67). Patients showed reduced HR-Acc (p < .006, d = 0.97) and HR-Vm coupling (p < .009, d = 0.91). Combining Vm and Acc to predict HR showed the strongest group separation (p < .002, d = 1.07). Discriminant analyses, based on the combination of Vm and Acc to predict HR, classified 77% of all participants correctly. In PD, HR-Acc coupling was inversely related to trait anxiety sensitivity, as well as tonic and phasic daytime anxiety. The novel method that was used demonstrates that anxiety in PD may reduce efficient long-term metabolic coupling. Metabolic decoupling may serve as physiological characteristic of PD and might aid diagnostics for PD and other anxiety disorders. This measure deserves further study in research on health consequences of anxiety and psychosocial stress.

Positive impact of IGF-1-coupled nanoparticles on the differentiation potential of human chondrocytes cultured on collagen scaffolds.

PurposeIn the present study, silica nanoparticles (sNP) coupled with insulin-like growth factor 1 (IGF-1) were loaded on a collagen-based scaffold intended for cartilage repair, and the influence on the viability, proliferation, and differentiation potential of human primary articular chondrocytes was examined.MethodsHuman chondrocytes were isolated from the hyaline cartilage of patients (n=4, female, mean age: 73±5.1 years) undergoing primary total knee joint replacement. Cells were dedifferentiated and then cultivated on a bioresorbable collagen matrix supplemented with fluorescent sNP coupled with IGF-1 (sNP–IGF-1). After 3, 7, and 14 days of cultivation, cell viability and integrity into the collagen scaffold as well as metabolic cell activity and synthesis rate of matrix proteins (collagen type I and II) were analyzed.ResultsThe number of vital cells increased over 14 days of cultivation, and the cells were able to infiltrate the collagen matrix (up to 120 μm by day 7). Chondrocytes cultured on the collagen scaffold supplemented with sNP–IGF-1 showed an increase in metabolic activity (5.98-fold), and reduced collagen type I (1.58-fold), but significantly increased collagen type II expression levels (1.53-fold; P=0.02) after 7 days of cultivation compared to 3 days. In contrast, chondrocytes grown in a monolayer on plastic supplemented with sNP-IGF-1 had significantly lower metabolic activity (1.32-fold; P=0.007), a consistent amount of collagen type I, and significantly reduced collagen type II protein expression (1.86-fold; P=0.001) after 7 days compared to 3 days.ConclusionCollagen-based scaffolds enriched with growth factors, such as IGF-1 coupled to nanoparticles, represent an improved therapeutic intervention for the targeted and controlled treatment of articular cartilage lesions.

Fine structure of kinetin-treated protonema and kinetin-induced gametophore buds in Funaria hygrometrica

Besides occasional hypertrophy of grana and disintegration of stroma thylakoids occurring in some chloroplasts, no significant changes were found in ultrastructure of typical protonema cells treated for six days with kinetin. On the other hand, the fine structure of cells in kinetin--induced gametophore buds differed much from that of the protonema cells and showed characteristics of cells of with high metabolic activity and high division rates. The results indicate that cytokinins enhance development and differentiation in the protonema by activating only some of its cells, whereas the others remain unchanged or show symptoms of destruction and ageing. This is supported by the fact that in the presence of chloramphenicol, which prevents bud induction, kinetin acts synergistically with the inhibitor in producing degeneration and destruction of chloroplasts.

Embryonic development and rates of metabolic activity in early and late hatching eggs of the major malaria vector Anopheles gambiae.

Anopheles gambiae eggs generally hatch at the completion of embryo development; two-three days post oviposition. However, staggered or delayed hatching has been observed whereby a single batch of eggs shows marked variation in time-to-hatch, with some eggs hatching 18 days post oviposition or later. The mechanism enabling delayed hatch has not been clearly elucidated but is likely mediated by environmental and genetic factors that either induce diapause or slow embryo development. This study aimed to compare metabolic activity and embryonic development between eggs collected from sub-colonies of the baseline Anopheles gambiae GAH colony previously selected for early or late time-to-hatch. Egg batches from early and late hatch sub-colonies as well as from the baseline colony were monitored for hatching. For both time-to-hatch selected sub-colonies and the baseline colony the majority of eggs hatched on day two post oviposition. Nevertheless, eggs produced by the late hatch sub-colony showed a significantly longer mean time to hatch than those produced by the early hatch sub-colony. The overall proportions that hatched were similar for all egg batches. CO2 output between eggs from early and late hatch sub-colonies showed significant differences only at 3 and 7 days post oviposition where eggs from the early hatch and the late hatch sub-colony were more metabolically active, respectively. No qualitative differences were observed in embryo development between the sub-colonies. It is concluded that all viable embryos develop to maturity at the same rate and that a small proportion then enter a state of diapause enabling them to hatch later. As it has previously been shown that it is possible to at least partially select for late hatch, this characteristic is likely to involve genetic as well as environmental factors. Delayed hatching in An. gambiae is likely an adaptation to maximise reproductive output despite the increased risk of desiccation in an unstable aquatic environment.

The short- and long-term effects of environmental conditions on anaerobic methane oxidation coupled to nitrite reduction

Anaerobic oxidation of methane coupled to nitrite reduction (n-damo) plays an important role in global carbon and nitrogen cycles and also is a potential bioprocess in wastewater treatment. In this work, the effects of environmental conditions – temperature, pH and salinity – on the metabolic activity and growth rate of n-damo bacteria were investigated by short-term batch test and long-term bacterial incubation. Quantitative PCR and 16S rRNA and pmoA gene sequencing were applied to detect the microbial community in the long-term incubation. The results indicated that all the three environmental factors significantly affected the metabolic activity and growth rate of n-damo bacteria and the optimum temperature, pH and salinity were 35 °C, 7.6 and 0 g NaCl L−1, respectively. Notably, salinity adaption of n-damo bacteria was first observed under salinity stress of 20 g NaCl L−1. It's predicted that n-damo process might occur in saline environments and future work could focus on this.

Functional characteristics of yeast cells in nutrient aqueous solution enriched with ortho-H2O isomers

It has been experimentally established that cultivation of yeast cells in depleted, dietary or normal nutrient aqueous solutions enriched with ortho-H2O spin isomers is accompanied by an increase in the amount of carbon dioxide produced by the cells and an increase in their biomass. It has been revealed that the rate of metabolic processes and biological activity depends on the quality of nutrition and enhances in time in both nutrient solutions. In contrast, the reproductive function and the rate of cell division are insusceptible to the components of nutrition, but intensified in a solution enriched with ortho-H2O similar to retardation of aging. The observed effects are discussed in assumption that an increase of a portion of ortho-H2O molecules occurs in the neighborhood of water channels in the cell membrane that let through only monomers of H2O and determine the rate of metabolic processes.

Mechanisms of paracrine cardioprotection by cord blood mesenchymal stromal cells

Among the mechanisms by which somatic stem cells may improve left ventricular function in ischaemic heart disease are pro-survival stimuli mediated by secreted factors. This phenomenon is frequently referred to, but remains poorly understood. We therefore investigated the non-regenerative cardioprotective effects of cord blood mesenchymal stromal cells (CBMSCs) in vitro and sought to identify relevant intracellular signalling pathways. Conditioned medium from CBMSCs and fibroblasts was prepared, and secreted factors were analysed by Luminex(®) immunobead assay. Murine cardiomyocyte-derived HL-1 cells were subjected to simulated ischaemia by glucose and serum deprivation and hypoxia in CBMSC-conditioned or cell-free control medium or in medium conditioned by foreskin fibroblasts. The proportions of vital, apoptotic and necrotic cells (poly-caspase activity, annexin V and ethidium homodimer-III staining) were quantified using a high-content imaging system. Metabolic activity and proliferation rate were determined via 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium and 5-bromo-2-deoxyuridine assays. Phosphorylation of Akt, extracellular-signal-regulated kinase (ERK)1/2, signal transducer and activator of transcription 3 (STAT3) and glycogen synthase kinase 3β was determined by western blot, and experiments were repeated in the presence of specific small-molecule inhibitors (Wortmannin, UO126 and Stattic). CBMSC medium reduced the proportion of dead HL-1 cardiomyocytes from 39 ± 3 to 28 ± 1% (P < 0.05) and the rate of late apoptotic cells to 68 ± 2% of that in control medium (P < 0.001). Metabolic activity was increased by 12 ± 1% compared with control (P < 0.05), while in fibroblast medium it was not (5 ± 2%, P = 1). This was associated with increased phosphorylation of Akt (2-fold, P < 0.05), ERK1/2 (3-fold, P < 0.01) and STAT3 (12-fold, P < 0.001). Combined blocking of the phosphatidylinositol-4,5-bisphosphate 3-kinase/Akt and mitogen-activated protein kinase/ERK signalling abolished the protective CBMSC effect, while blocking the pathways individually had no effect. Inhibition of STAT3 phosphorylation drastically lowered HL-1 cell viability in control medium, but not in medium conditioned by CBMSCs. The factors released by CBMSCs protect cardiomyocyte-like HL-1 cells from simulated ischaemia more than those released from fibroblasts. While CBMSC-triggered Akt and ERK1/2 activation provides protection in a compensatory manner, STAT3 is crucial for cardiomyocyte survival in ischaemia, but is not a key mediator of cytoprotective stem cell actions.

Uremic toxins impair human bone marrow-derived mesenchymal stem cells functionality in vitro

Mesenchymal stem cells (MSCs) are becoming therapeutic agents of interest in many areas of medicine, including renal diseases and kidney transplantations. However, the effect of uremia on cell properties is still unclear. Therefore, we examined the in vitro influence of uremic toxins, p-cresol (PC) and indoxyl sulfate (IS), on human bone marrow-derived MSC functionality. Cultured MSCs were treated with PC and IS at concentrations corresponding to subsequent stages of chronic kidney disease. Cell viability was characterized by metabolic activity (MTT assay) and proliferation rate (BrdU assay). Apoptosis (Annexin V test) and cell membrane damage (LDH assay) were also tested. MSC secretory properties were determined by measuring cytokine/growth factor levels in media from toxin-treated cells (ELISA). Uremic concentrations of PC and IS resulted in significant inhibition of MSC metabolic activity and proliferation. Toxins did not induce apoptosis, but damaged cell membranes. MSC paracrine activity was also altered – a decrease of VEGF and TGF-β1 levels and an increase in IGF-1 and IL-8 secretion was detected. Presented data indicate a negative influence of uremic toxins on functional characteristics of human bone marrow-derived MSCs. Therefore, their use as autologous therapeutic agents for kidney disease may be questionable and requires further investigations. The observed phenomenon may be attributable to many other MSC therapies, because of the high prevalence of chronic kidney disease in adult population.

Direct enzyme-substrate affinity determination by real-time hyperpolarized (13)C-MRS.

A specialized kinetic analysis of real-time hyperpolarized [1,1,2,2-D4, 1-(13)C]choline (13)C-magnetic resonance spectroscopy enabled the determination of initial rates of metabolic enzyme activity (choline oxidase), enzyme-substrate affinity (Km), and inhibition. In a clinical MRI scanner, metabolite levels lower than 16 μM were detected at a temporal resolution of 1 s.

Antioxidant Protective Effects of Mushroom Metabolites

Global industrialization and agricultural development are related to the release of various pollutants into the environment including huge amounts of free radicals, which are associated with the occurrence of various destructive diseases and disorders. Due to different rates of metabolic activity and oxygen consumption, organisms, organs and tissues have distinct protective antioxidant systems and defence mechanisms. Although numerous synthetic antioxidants can improve defence system capacity, because of their toxic and mutagenic effects as well as rising awareness of a healthy lifestyle, preference is given to natural antioxidants. Therefore, studies of metabolites of various fungal species and their activity are currently increasing. Antioxidant features of numerous compounds isolated from mushrooms, such as phenolic compounds, vitamins, polysaccharides, peptides, proteins, organic acids, carotenoids, alkaloids, and nucleotides, have been reported. Chemical composition and antioxidant potential of mushrooms are highly-dependent on species, habitat, phase of life cycle (mycelium, young or mature fruiting body), method of processing, extraction solvent, and extract dose. Although around only 5% of fungal species have been well studied and thousands more species have potential benefit for mankind, reports on the antioxidant potentials of fungi are already numerous and these are the subject of this review.

Essential Role of the Cytochrome P450 Enzyme CYP2A5 in Olfactory Mucosal Toxicity of Naphthalene

Naphthalene (NA), a ubiquitous environmental pollutant that can cause pulmonary and nasal toxicity in laboratory animals, requires cytochrome P450 (P450)-mediated metabolic activation to cause toxicity. Our recent study using a Cyp2f2-null mouse showed that CYP2F2 plays an essential role in NA-induced lung toxicity, but not in NA-induced nasal toxicity. The aim of this study was to determine whether mouse CYP2A5, abundantly expressed in nasal olfactory mucosa (OM) and the liver, but less in the lung, plays a major role in the bioactivation and toxicity of NA in the OM. We found, by comparing Cyp2a5-null and wild-type (WT) mice, that the loss of CYP2A5 expression led to substantial decreases in rates of NA metabolic activation by OM microsomes. The loss of CYP2A5 did not cause changes in systemic clearance of NA (at 200 mg/kg, i.p.). However, the Cyp2a5-null mice were much more resistant than were WT mice to NA-induced nasal toxicity (although not lung toxicity), when examined at 24 hours after NA dosing (at 200 mg/kg, i.p.), or to NA-induced depletion of total nonprotein sulfhydryl in the OM (although not in the lung), examined at 2 hours after dosing. Thus, mouse CYP2A5 plays an essential role in the bioactivation and toxicity of NA in the OM, but not in the lung. Our findings further illustrate the tissue-specific nature of the role of individual P450 enzymes in xenobiotic toxicity, and provide the basis for a more reliable assessment of the potential risks of NA nasal toxicity in humans.

Hexamitiasis leads to lower metabolic rates in rainbow trout Oncorhynchus mykiss (Walbaum) juveniles

This study assessed the effects of Hexamita salmonis (Moore) on metabolism of rainbow trout Oncorhynchus mykiss (Walbaum) and its effect on the host's susceptibility to infectious pancreatic necrosis virus (IPNV) after antiparasitic treatment. Rainbow trout naturally infected with H. salmonis were treated with 10 mg metronidazole kg fish(-1) per day, and their physiological recovery was assessed through measuring resting metabolism on the 7th, 14th, 21st and 28th day after treatment. In addition, we exposed the naïve fish to H. salmonis and measured the resting metabolism (oxygen consumption as mg O2 kg(-1) per hour) on the 10th, 20th and 30th day after the exposure to assess the variation in metabolic rates after infection. Significantly lower rates of metabolic activity (P < 0.05) were anticipated 20 days after infection with H. salmonis compared with the fish infected with H. salmonis for 10 days or with the parasite-free fish. Similarly, the treated fish needed about 20 days to fully recover from hexamitiasis. The susceptibility of rainbow trout to IPNV remained unchanged in the presence of H. salmonis. Weight loss was significantly higher (P < 0.05) in infected than that in the parasite-free fish. Fish should be examined regularly for H. salmonis and treated immediately whether found to prevent economic losses and excessive size variation.

Bio-inspired fabrication of fibroin cryogels from the muga silkworm Antheraea assamensis for liver tissue engineering

Conventional scaffold fabrication techniques result in narrow pore architectures causing a limited interconnectivity and use of porogens, which affects the bio- or cyto-compatibility. To ameliorate this, cryogels are immensely explored due to their macro-porous nature, ease in fabrication, using ice crystals as porogens, the shape property, easy reproducibility and cost-effective fabrication technique. Cryogels in the present study are prepared from nonmulberry Indian muga silk gland protein fibroin of Antheraea assamensis using two different fabrication temperatures (−20 and −80 °C). Anionic surfactant sodium dodecyl sulfate is used to solubilize fibroin, which in turn facilitates gelation by accelerating the ß-sheet formation. Ethanol is employed to stabilize the 3D network and induces bimodal porosity. The gels thus formed demonstrate increased ß-sheet content (FTIR) and a considerable effect of pre-freezing temperatures on 3D micro-architectures. The cryogels are capable of absorbing large amounts of water and withstanding mechanical compression without structure deformation. Further, cell impregnated cryogels well support the viability of human hepatocarcinoma cells (live/dead assay). The formation of cellular aggregates (confocal laser and scanning electron microscope), derivation in metabolic activity and proliferation rate are obtained in constructs fabricated at different temperatures. In summary, the present work reveals promising insights in the development of a biomimetic functional template for biomedical therapeutics and liver tissue engineering.

CO2 enhances effects of hypoxia on mortality, development, and gene expression in cowpea bruchid, Callosobruchus maculatus

Modified atmosphere based on lack of O2 offers a safe, residue-free alternative to chemical fumigants for pest control in stored grains. In this study, we intended to determine whether elevated CO2 (at a biologically achievable level) has an enhanced suppressive effect over low O2 atmosphere alone on the cowpea bruchid (Callosobruchus maculatus), a storage pest of cowpea and other legumes. Experiments were performed under two modified atmospheric conditions, (1) 2% O2+18% CO2+80% N2 and (2) 2% O2+98% N2. Both hypoxic environments significantly affected the development and survival of all insect developmental stages. Eggs were most vulnerable to hypoxia, particularly at the early stage (4–6h old), surviving only up to a maximum of 2 days in both treatments. These were followed by adults, pupae and larvae, in order of decreasing susceptibility. The 3rd and 4th instar larvae were most resilient to hypoxia and could survive up to 20days of low O2. The presence of 18% CO2 significantly increased the mortality of adults, the later stage of eggs, as well as 1st and 4th instar larvae caused by hypoxia. However, the surviving insects exhibited faster development, evidenced by their earlier emergence from cowpea seeds compared to those without CO2. One interesting observation was the frequent, premature opening of the emergence windows in the 4th instar larvae when CO2 was involved. This phenomenon was not observed at all in insects stressed by low O2 alone. Differential expression profiling of metabolic genes and proteolytic activity of midgut digestive enzymes suggested that the rate of metabolic activity could contribute in part to the difference in insect development and survival under hypoxia in the presence and absence of CO2.

Skeletal Muscle Atrophy in Bioengineered Skeletal Muscle: A New Model System

Skeletal muscle atrophy has been well characterized in various animal models, and while certain pathways that lead to disuse atrophy and its associated functional deficits have been well studied, available drugs to counteract these deficiencies are limited. An ex vivo tissue-engineered skeletal muscle offers a unique opportunity to study skeletal muscle physiology in a controlled in vitro setting. Primary mouse myoblasts isolated from adult muscle were tissue engineered into bioartificial muscles (BAMs) containing hundreds of aligned postmitotic muscle fibers expressing sarcomeric proteins. When electrically stimulated, BAMs generated measureable active forces within 2-3 days of formation. The maximum isometric tetanic force (Po) increased for ∼3 weeks to 2587±502 μN/BAM and was maintained at this level for greater than 80 days. When BAMs were reduced in length by 25% to 50%, muscle atrophy occurred in as little as 6 days. Length reduction resulted in significant decreases in Po (50.4%), mean myofiber cross-sectional area (21.7%), total protein synthesis rate (22.0%), and noncollagenous protein content (6.9%). No significant changes occurred in either the total metabolic activity or protein degradation rates. This study is the first in vitro demonstration that length reduction alone can induce skeletal muscle atrophy, and establishes a novel in vitro model for the study of skeletal muscle atrophy.

Design of optimized hypoxia-activated prodrugs using pharmacokinetic/pharmacodynamic modeling.

Hypoxia contributes to resistance of tumors to some cytotoxic drugs and to radiotherapy, but can in principle be exploited with hypoxia-activated prodrugs (HAP). HAP in clinical development fall into two broad groups. Class I HAP (like the benzotriazine N-oxides tirapazamine and SN30000), are activated under relatively mild hypoxia. In contrast, Class II HAP (such as the nitro compounds PR-104A or TH-302) are maximally activated only under extreme hypoxia, but their active metabolites (effectors) diffuse to cells at intermediate O2 and thus also eliminate moderately hypoxic cells. Here, we use a spatially resolved pharmacokinetic/pharmacodynamic (SR-PK/PD) model to compare these two strategies and to identify the features required in an optimal Class II HAP. The model uses a Green’s function approach to calculate spatial and longitudinal gradients of O2, prodrug, and effector concentrations, and resulting killing in a digitized 3D tumor microregion to estimate activity as monotherapy and in combination with radiotherapy. An analogous model for a normal tissue with mild hypoxia and short intervessel distances (based on a cremaster muscle microvessel network) was used to estimate tumor selectivity of cell killing. This showed that Class II HAP offer advantages over Class I including higher tumor selectivity and greater freedom to vary prodrug diffusibility and rate of metabolic activation. The model suggests that the largest gains in class II HAP antitumor activity could be realized by optimizing effector stability and prodrug activation rates. We also use the model to show that diffusion of effector into blood vessels is unlikely to materially increase systemic exposure for realistic tumor burdens and effector clearances. However, we show that the tumor selectivity achievable by hypoxia-dependent prodrug activation alone is limited if dose-limiting normal tissues are even mildly hypoxic.