Evaluating the effects of giraffe skin disease and wire snare wounds on the gaits of free-ranging Nubian giraffe.

Giraffe skin disease (GSD), a condition that results in superficial lesions in certain giraffe (Giraffa spp.) populations, has emerged as a potential conservation threat. Preliminary findings suggested that individuals with GSD lesions move with greater difficulty which may in turn reduce their foraging efficiency or make them more vulnerable to predation. A current known threat to some giraffe populations is their mortality associated with entrapment in wire snares, and the morbidity and potential locomotor deficiencies associated with wounds acquired from snares. The goal of our study was to quantify the locomotor kinematics of free-ranging Nubian giraffe (G. camelopardalis camelopardalis) in Murchison Falls National Park (MFNP), Uganda, and compare spatiotemporal limb and neck angle kinematics of healthy giraffe to those of giraffe with GSD lesions, snare wounds, and both GSD lesions and snare wounds. The presence of GSD lesions did not significantly affect spatiotemporal limb kinematic parameters. This finding is potentially because lesions were located primarily on the necks of Nubian giraffe in MFNP. The kinematic parameters of individuals with snare wounds differed from those of healthy individuals, resulting in significantly shorter stride lengths, reduced speed, lower limb phase values, and increased gait asymmetry. Neck angle kinematic parameters did not differ among giraffe categories, which suggests that GSD neck lesions do not impair normal neck movements and range of motion during walking. Overall, MFNP giraffe locomotor patterns are largely conservative between healthy individuals and those with GSD, while individuals with snare wounds showed more discernible kinematic adjustments consistent with unilateral limb injuries. Additional studies are recommended to assess spatiotemporal limb kinematics of giraffe at sites where lesions are found predominantly on the limbs to better assess the potential significance of GSD on their locomotion.

Boundary Zonal Flow in Rotating Turbulent Rayleigh-Bénard Convection.

For rapidly rotating turbulent Rayleigh-Bénard convection in a slender cylindrical cell, experiments and direct numerical simulations reveal a boundary zonal flow (BZF) that replaces the classical large-scale circulation. The BZF is located near the vertical side wall and enables enhanced heat transport there. Although the azimuthal velocity of the BZF is cyclonic (in the rotating frame), the temperature is an anticyclonic traveling wave of mode one, whose signature is a bimodal temperature distribution near the radial boundary. The BZF width is found to scale like Ra^{1/4}Ek^{2/3} where the Ekman number Ek decreases with increasing rotation rate.

Twente mass and heat transfer water tunnel: Temperature controlled turbulent multiphase channel flow with heat and mass transfer.

A new vertical water tunnel with global temperature control and the possibility for bubble and local heat and mass injection has been designed and constructed. The new facility offers the possibility to accurately study heat and mass transfer in turbulent multiphase flow (gas volume fraction up to 8%) with a Reynolds-number range from 1.5 × 104 to 3 × 105 in the case of water at room temperature. The tunnel is made of high-grade stainless steel permitting the use of salt solutions in excess of 15% mass fraction. The tunnel has a volume of 300 l. The tunnel has three interchangeable measurement sections of 1 m height but with different cross sections (0.3 × 0.04 m2, 0.3 × 0.06 m2, and 0.3 × 0.08 m2). The glass vertical measurement sections allow for optical access to the flow, enabling techniques such as laser Doppler anemometry, particle image velocimetry, particle tracking velocimetry, and laser-induced fluorescent imaging. Local sensors can be introduced from the top and can be traversed using a built-in traverse system, allowing, for example, local temperature, hot-wire, or local phase measurements. Combined with simultaneous velocity measurements, the local heat flux in single phase and two phase turbulent flows can thus be studied quantitatively and precisely.

Different properties of skin of different body sites: The root of keloid formation?

The purpose of this study was to examine extracellular matrix composition, vascularization, and immune cell population of skin sites prone to keloid formation. Keloids remain a complex problem, posing esthetical as well as functional difficulties for those affected. These scars tend to develop at anatomic sites of preference. Mechanical properties of skin vary with anatomic location and depend largely on extracellular matrix composition. These differences in extracellular matrix composition, but also vascularization and resident immune cell populations might play a role in the mechanism of keloid formation. To examine this hypothesis, skin samples of several anatomic locations were taken from 24 human donors within zero to 36 hours after they had deceased. Collagen content and cross-links were determined through high-performance liquid chromatography. The expression of several genes, involved in extracellular matrix production and degradation, was measured by means of real-time PCR. (Immuno)histochemistry was performed to detect fibroblasts, collagen, elastin, blood vessels, Langerhans cells, and macrophages. Properties of skin of keloid predilections sites were compared to properties of skin from other locations (nonpredilection sites [NPS]). The results indicated that there are site specific variations in extracellular matrix properties (collagen and cross-links) as well as macrophage numbers. Moreover, predilection sites (PS) for keloid formation contain larger amounts of collagen compared to NPS, but decreased numbers of macrophages, in particular classically activated CD40 positive macrophages. In conclusion, the altered (histological, protein, and genetic) properties of skin of keloid PS may cause a predisposition for and contribute to keloid formation.

Inhibited early immunologic response is associated with hypertrophic scarring.

This study aimed to examine changes in the inflammatory response in early hypertrophic compared to normal wound healing. The immune system is thought to be involved in hypertrophic scar formation. However, the exact mechanism and time of onset of the derailment remain unknown. In a prospective observational study, skin biopsies were taken directly postwounding and 3 hours later from patients who had elective cardiothoracic surgery. The skin biopsies were analysed for mRNA, proteins and cells involved in the early inflammatory phase of wound healing. The endpoint was scar outcome (hypertrophic (HTS) or normal (NTS)) at one year after surgery. There were significant differences between the NTS and HTS groups regarding the fold changes of mRNA expression of P-selectin during surgery. Postoperative skin concentrations of inflammatory proteins IL-6, IL-8 and CCL2 were significantly lower in the HTS compared to the NTS group. Also, a trend of higher pre-operative M2 macrophage numbers was observed in the HTS group. Neutrophil numbers increased equally during surgery in both groups. The increase of P-selectin mRNA in hypertrophic wound healing could affect leucocyte migration. The decreased concentrations of inflammatory proteins in hypertrophic wound healing indicate a reduced inflammatory response, which has consequences for the treatment of hypertrophic scarring during the early inflammatory phase. In a conclusion, alterations of wound healing associated with hypertrophic scarring are visible as early as 3 hours postwounding and include a reduced rather than increased inflammatory protein response.

Bone Sarcomas in Pediatrics: Progress in Our Understanding of Tumor Biology and Implications for Therapy.

The pediatric bone sarcomas osteosarcoma and Ewing sarcoma represent a tremendous challenge for the clinician. Though less common than acute lymphoblastic leukemia or brain tumors, these aggressive cancers account for a disproportionate amount of the cancer morbidity and mortality in children, and have seen few advances in survival in the past decade, despite many large, complicated, and expensive trials of various chemotherapy combinations. To improve the outcomes of children with bone sarcomas, a better understanding of the biology of these cancers is needed, together with informed use of targeted therapies that exploit the unique biology of each disease. Here we summarize the current state of knowledge regarding the contribution of receptor tyrosine kinases, intracellular signaling pathways, bone biology and physiology, the immune system, and the tumor microenvironment in promoting and maintaining the malignant phenotype. These observations are coupled with a review of the therapies that target each of these mechanisms, focusing on recent or ongoing clinical trials if such information is available. It is our hope that, by better understanding the biology of osteosarcoma and Ewing sarcoma, rational combination therapies can be designed and systematically tested, leading to improved outcomes for a group of children who desperately need them.

The narrow-spectrum HDAC inhibitor entinostat enhances NKG2D expression without NK cell toxicity, leading to enhanced recognition of cancer cells.

PURPOSE: Natural killer (NK) cell cytotoxicity correlates with the ligation of activating receptors (e.g., NKG2D) by their ligands (e.g., MHC class I-related chains [MIC] A and B) on target cells. Histone deacetylase inhibitors (HDACi) at high concentrations inhibit tumor growth and can increase NKG2D ligand expression on tumor targets, but are widely regarded as toxic to NK cells. METHODS: We investigated the mechanism of entinostat, a benzamide-derivative narrow-spectrum HDACi, in augmenting the cytotoxicity of NK cells against human colon carcinoma and sarcoma by assessing gene and protein expression, histone acetylation, and cytotoxicity in in vitro and murine models. RESULTS: We observed that entinostat dose- and time-dependent increase in MIC expression in tumor targets and NKG2D in primary human NK cells, both correlating with increased acetylated histone 3 (AcH3) binding to associated promoters. Entinostat pretreatment of colon carcinoma and sarcoma cells, NK cells, or both led to enhanced overall cytotoxicity in vitro, which was reversed by NKG2D blockade, and inhibited growth of tumor xenografts. Lastly, we showed decreased expression of MICA and ULBP2 transcription in primary human osteosarcoma. CONCLUSIONS: Entinostat enhances NK cell killing of cancer cells through upregulation of both NKG2D and its ligands, suggesting an attractive approach for augmenting NK cell immunotherapy of solid tumors such as colon carcinoma and sarcomas.

New, tolerable γ-secretase inhibitor takes desmoid down a notch.

A phase I trial of PF-03084014, an oral reversible γ-secretase inhibitor, in solid tumor malignancies shows drug tolerability in patients. Evidence of Notch pathway inhibition was demonstrated in peripheral blood. A surprisingly high rate of response was seen in desmoid tumors, a rare but sometimes locally aggressive sarcoma.

Understanding the role of Notch in osteosarcoma.

The Notch pathway has been described as an oncogene in osteosarcoma, but the myriad functions of all the members of this complex signaling pathway, both in malignant cells and nonmalignant components of tumors, make it more difficult to define Notch as simply an oncogene or a tumor suppressor. The cell-autonomous behaviors caused by Notch pathway manipulation may vary between cell lines but can include changes in proliferation, migration, invasiveness, oxidative stress resistance, and expression of markers associated with stemness or tumor-initiating cells. Beyond these roles, Notch signaling also plays a vital role in regulating tumor angiogenesis and vasculogenesis, which are vital aspects of osteosarcoma growth and behavior in vivo. Further, osteosarcoma cells themselves express relatively low levels of Notch ligand, making it likely that nonmalignant cells, especially endothelial cells and pericytes, are the major source of Notch activation in osteosarcoma tumors in vivo and in patients. As a result, Notch pathway expression is not expected to be uniform across a tumor but likely to be highest in those areas immediately adjacent to blood vessels. Therapeutic targeting of the Notch pathway is likewise expected to be complicated. Most pharmacologic approaches thus far have focused on inhibition of gamma secretase, a protease of the presenilin complex. This enzyme, however, has numerous other target proteins that would be expected to affect osteosarcoma behavior, including CD44, the WNT/ß-catenin pathway, and Her-4. In addition, Notch plays a vital role in tissue and organ homeostasis in numerous systems, and toxicities, especially GI intolerance, have limited the effectiveness of gamma secretase inhibitors. New approaches are in development, and the downstream targets of Notch pathway signaling also may turn out to be good targets for therapy. In summary, a full understanding of the complex functions of Notch in osteosarcoma is only now unfolding, and this deeper knowledge will help position the field to better utilize novel therapies as they are developed.

Developmental pathways hijacked by osteosarcoma.

Cancer of any type often can be described by an arrest, alteration or disruption in the normal development of a tissue or organ, and understanding of the normal counterpart's development can aid in understanding the malignant state. This is certainly true for osteosarcoma and the normal developmental pathways that guide osteoblast development that are changed in the genesis of osteogenic sarcoma. A carefully regulated crescendo-decrescendo expression of RUNX2 accompanies the transition from mesenchymal stem cell to immature osteoblast to mature osteoblast. This pivotal role is controlled by several pathways, including bone morphogenic protein (BMP), Wnt/ß-catenin, fibroblast growth factor (FGF), and protein kinase C (PKC). The HIPPO pathway and its downstream target YAP help to regulate proliferation of immature osteoblasts and their maturation into non-proliferating mature osteoblasts. This pathway also helps regulate expression of the mature osteoblast protein osteocalcin. YAP also regulates expression of MT1-MMP, a membrane-bound matrix metalloprotease responsible for remodeling the extracellular matrix surrounding the osteoblasts. YAP, in turn, can be regulated by the ERBB family protein Her-4. Osteosarcoma may be thought of as a cell held at the immature osteoblast stage, retaining some of the characteristics of that developmental stage. Disruptions of several of these pathways have been described in osteosarcoma, including BMP, Wnt/b-catenin, RUNX2, HIPPO/YAP, and Her-4. Further, PKC can be activated by several receptor tyrosine kinases implicated in osteosarcoma, including the ERBB family (EGFR, Her-2 and Her-4 in osteosarcoma), IGF1R, FGF, and others. Understanding these functions may aid in the understanding the mechanisms underpinning clinical observations in osteosarcoma, including both the lytic and blastic phenotypes of tumors, the invasiveness of the disease, and the tendency for treated tumors to ossify rather than shrink. Through a better understanding of the relationship between normal osteoblast development and osteosarcoma, we may gain insights into novel therapeutic avenues and improved outcomes.

Logarithmic spatial variations and universal f-1 power spectra of temperature fluctuations in turbulent Rayleigh-Bénard convection.

We report measurements of the temperature variance σ(2)(z,r) and frequency power spectrum P(f,z,r) (z is the distance from the sample bottom and r the radial coordinate) in turbulent Rayleigh-Bénard convection (RBC) for Rayleigh numbers Ra = 1.6 × 10(13) and 1.1 × 10(15) and for a Prandtl number Pr ≃ 0.8 for a sample with a height L = 224 cm and aspect ratio D/L=0.50 (D is the diameter). For z/L ≲ 0.1 σ(2)(z,r) was consistent with a logarithmic dependence on z, and there was a universal (independent of Ra, r, and z) normalized spectrum which, for 0.02 ≲ fτ(0) ≲ 0.2, had the form P(fτ(0)) = P(0)(fτ(0))(-1) with P(0) = 0.208 ± 0.008 a universal constant. Here τ(0) = sqrt[2R] where R is the radius of curvature of the temperature autocorrelation function C(τ) at τ = 0. For z/L ≃ 0.5 the measurements yielded P(fτ(0))∼(fτ(0))(-α) with α in the range from 3/2 to 5/3. All the results are similar to those for velocity fluctuations in shear flows at sufficiently large Reynolds numbers, suggesting the possibility of an analogy between the flows that is yet to be determined in detail.

Epidemiology of toxoplasmosis in white tailed deer (Odocoileus virginianus): occurrence, congenital transmission, correlates of infection, isolation, and genetic characterization of Toxoplasma gondii.

The prevalence of Toxoplasma gondii in white tailed deer (WTD) in the USA is high but little is known of the epidemiology of toxoplasmosis in this host. In the present study, we compared T. gondii seroprevalence from 749 WTD collected in 2012 and 2013 from a Metropolitan Park in Ohio and 487 WTD deer shot in Minnesota during 2008, 2009, and 2010. Serum samples were tested for antibodies to T. gondii by the modified agglutination test (cut-off titer, 25). Additionally myocardial samples from 123 seropositive WTD from Ohio were digested in pepsin and the digests were bioassayed for the isolation of T. gondii. Furthermore, to estimate transplacental rate of transmission, brains from 155 fetuses (included twins) from 148 deer from Minnesota were bioassayed in mice for the isolation of viable T. gondii. Seroprevalence of T. gondii varied with the year of collection, geography, and the age of deer. Of the Ohio deer sampled in 2012 and 2013 seroprevalences for the two years were similar (73.4% and 75.7%, respectively); remarkably 150 (66.1%) of 227 deer of <1 year of age were seropositive. Of the Minnesota deer, seroprevalence was lowest for the year 2008 (14.8%, 26/175) versus 2009 (27.7%, 59/213), and 2010 (25.2%, 25/99), thought to be related to environmental temperatures. Viable T. gondii was isolated in mice from the myocardium of four WTD from Ohio, and brain of one WTD fetus from Minnesota. Tachyzoites from infected mouse tissues were further propagated in cell culture. The DNA isolated from culture-derived tachyzoites of these five T. gondii isolates was characterized using 11 PCR-RFLP markers (SAG1, 5'- and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). Four genotypes were found, including ToxoDB genotype no. 1 (Type II), no. 2 (Type III), no. 3 (Type II variant) and no. 146. Results indicate fluctuating seroprevalence, probably related to weather and warrant further epidemiological studies.

Implementing a low-starch biscuit-free diet in zoo gorillas: the impact on health.

In the wild, western lowland gorillas consume a diet high in fiber and low in caloric density. In contrast, many gorillas in zoos consume a diet that is high-calorie and low in fiber. Some items commonly used in captive gorilla diets contain high levels of starch and sugars, which are minimal in the natural diet of gorillas. There is a growing concern that captive gorillas may qualify as obese. Furthermore, the leading cause of death for adult male gorillas in zoos is heart disease. In humans, a diet that is high in simple carbohydrates is associated with both obesity and the incidence of heart disease. In response to these issues, we implemented a biscuit-free diet (free of biscuits and low in fruit) and measured serum biomarkers of obesity and insulin resistance pre- and post-diet change at three institutions: North Carolina Zoological Garden, Cleveland Metroparks Zoo, and Columbus Zoo and Aquarium. We also added a resistant starch supplement to gorilla diets at two of the above institutions. We anticipated that these diet changes would positively affect biomarkers of obesity and insulin resistance. Both diet manipulations led to a reduction in insulin. Resistant starch also decreased overall serum cholesterol levels. Future research will examine these health changes in a greater number of individuals to determine if the results remain consistent with these preliminary findings.

Implementing a low-starch biscuit-free diet in zoo gorillas: the impact on behavior.

In the wild, western lowland gorillas travel long distances while foraging and consume a diet high in fiber and low in caloric density. In contrast, gorillas in zoos typically consume a diet that is low in fiber and calorically dense. Some items commonly used in captive gorilla diets contain high levels of starch and sugars, which are present at low levels in the natural diet of gorillas. Diet items high in simple carbohydrates are associated with obesity and heart disease in humans. Typical captive gorilla diets may also encourage undesirable behaviors. In response to these issues, we tested the behavioral impact of a diet that was biscuit-free, had low caloric density, and which was higher in volume at five institutions. We hypothesized that this diet change would reduce abnormal behaviors such as regurgitation and reingestion (R/R), decrease time spent inactive, and increase time spent feeding. The biscuit-free diet significantly reduced (and in the case of one zoo eliminated) R/R and may have reduced hair-plucking behavior. However, an increase in coprophagy was observed in many individuals following the diet change. The experimental diet caused a general increase in time the gorillas spent feeding, but this increase did not occur across all institutions and varied by individual. Interestingly, the overall time gorillas spent inactive actually increased with this diet change. Future research will examine these behavioral changes in a greater number of individuals to determine if the results remain consistent with these preliminary findings. Additionally, future research will examine the physiological impact of this diet change.

Understanding the Biology of Bone Sarcoma from Early Initiating Events through Late Events in Metastasis and Disease Progression.

The two most common primary bone malignancies, osteosarcoma (OS), and Ewing sarcoma (ES), are both aggressive, highly metastatic cancers that most often strike teens, though both can be found in younger children and adults. Despite distinct origins and pathogenesis, both diseases share several mechanisms of progression and metastasis, including neovascularization, invasion, anoikis resistance, chemoresistance, and evasion of the immune response. Some of these processes are well-studies in more common carcinoma models, and the observation from adult diseases may be readily applied to pediatric bone sarcomas. Neovascularization, which includes angiogenesis and vasculogenesis, is a clear example of a process that is likely to be similar between carcinomas and sarcomas, since the responding cells are the same in each case. Chemoresistance mechanisms also may be similar between other cancers and the bone sarcomas. Since OS and ES are mesenchymal in origin, the process of epithelial-to-mesenchymal transition is largely absent in bone sarcomas, necessitating different approaches to study progression and metastasis in these diseases. One process that is less well-studied in bone sarcomas is dormancy, which allows micrometastatic disease to remain viable but not growing in distant sites - typically the lungs - for months or years before renewing growth to become overt metastatic disease. By understanding the basic biology of these processes, novel therapeutic strategies may be developed that could improve survival in children with OS or ES.

Complications in the surgical treatment of pediatric melanoma.

PURPOSE: The purpose of this study was to characterize the complications associated with surgical treatment of pediatric melanoma. METHODS: We retrospectively reviewed all pediatric patients who received surgical treatment for melanoma at our institution between 1992 and 2010. We compared complications between three groups: wide local excision only (WLE), WLE and sentinel lymph node biopsy (SLNB), and WLE and completion lymph node dissection (CLND). RESULTS: One hundred twenty-five patients were identified: 37 patients received WLE only, 47 received WLE and SLNB, and 41 patients had WLE and CLND. Complication rates differed between the three groups: 19% in WLE, 11% in WLE+SLNB, and 39% in WLE+CLND (P=.006). The risk of complications was significantly lower among patients having WLE+SLNB versus WLE+CLND (OR 0.19, 95% CI 0.06-0.57, P=.0032). Lymphedema was a common complication with a higher incidence in the CLND group compared to the SLNB group (19.5% vs. 2.1%, P=.01). Complications were more frequent in inguinal compared to axillary dissections (52.0% vs. 17.1%, P=.006). CONCLUSIONS: In the surgical treatment of pediatric melanoma, the addition of a completion lymph node dissection significantly increases complication risk. Thus, it is critical to determine which patients truly benefit from this procedure.

UBE4B levels are correlated with clinical outcomes in neuroblastoma patients and with altered neuroblastoma cell proliferation and sensitivity to epidermal growth factor receptor inhibitors.

BACKGROUND: The UBE4B gene, which is located on chromosome 1p36, encodes a ubiquitin ligase that interacts with hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs), a protein involved in epidermal growth factor receptor (EGFR) trafficking, suggesting a link between EGFR trafficking and neuroblastoma pathogenesis. The authors analyzed the roles of UBE4B in the outcomes of patients with neuroblastoma and in neuroblastoma tumor cell proliferation, EGFR trafficking, and response to EGFR inhibition. METHODS: The association between UBE4B expression and the survival of patients with neuroblastoma was examined using available microarray data sets. UBE4B and EGFR protein levels were measured in patient tumor samples, EGFR degradation rates were measured in neuroblastoma cell lines, and the effects of UBE4B on neuroblastoma tumor cell growth were analyzed. The effects of the EGFR inhibitor cetuximab were examined in neuroblastoma cells that expressed wild-type and mutant UBE4B. RESULTS: Low UBE4B gene expression is associated with poor outcomes in patients with neuroblastoma. UBE4B overexpression reduced neuroblastoma tumor cell proliferation, and UBE4B expression was inversely related to EGFR expression in tumor samples. EGFR degradation rates correlated with cellular UBE4B levels. Enhanced expression of catalytically active UBE4B resulted in reduced sensitivity to EGFR inhibition. CONCLUSIONS: The current study demonstrates associations between UBE4B expression and the outcomes of patients with neuroblastoma and between UBE4B and EGFR expression in neuroblastoma tumor samples. Moreover, levels of UBE4B influence neuroblastoma tumor cell proliferation, EGFR degradation, and response to EGFR inhibition. These results suggest UBE4B-mediated growth factor receptor trafficking may contribute to the poor prognosis of patients who have neuroblastoma tumors with 1p36 deletions and that UBE4B expression may be a marker that can predict responses of neuroblastoma tumors to treatment.

A survey of diabetes prevalence in zoo-housed primates.

In humans, type II diabetes mellitus is a condition in which the pancreas is capable of producing insulin but cells do not appropriately respond to insulin with an uptake of glucose. While multiple factors are associated with type II diabetes in humans, a high calorie diet and limited exercise are significant risk factors for the development of this disease. Zoo primates, with relatively high caloric density diets and sedentary lifestyles, may experience similar conditions that could predispose them to the development of diabetes. We surveyed all Association of Zoos and Aquariums (AZA) facilities with primates in their collections to determine the prevalence of diabetes, diagnosis and treatment methods, and treatment outcomes. Nearly 30% of responding institutions reported at least one diabetic primate in their current collection. Although the majority of reported cases were in Old World Monkeys (51%), all major taxonomic groups were represented. Females represented nearly 80% of the diagnosed cases. A wide variety of diagnosing, monitoring, and treatment techniques were reported. It is clear from these results diabetes should be considered prominently in decisions relating to diet, weight and activity levels in zoo-housed primates, as well as discussions surrounding animal health and welfare.

Slow down to stay alive: HER4 protects against cellular stress and confers chemoresistance in neuroblastoma.

BACKGROUND: Neuroblastoma (NBL) is a common pediatric solid tumor, and outcomes for patients with advanced neuroblastoma remain poor despite extremely aggressive treatment. Chemotherapy resistance at relapse contributes heavily to treatment failure. The poor survival of patients with high-risk NBL prompted this investigation into novel treatment options with the objective of gaining a better understanding of resistance mechanisms. On the basis of previous work and on data from publicly available studies, the authors hypothesized that human epidermal growth factor receptor 4 (Her4) contributes to resistance. METHODS: Her4 expression was reduced with small-hairpin RNA (shRNA) to over express intracellular HER4, and the authors tested its impact on tumor cell survival under various culture conditions. The resulting changes in gene expression after HER4 knockdown were measured by using a messenger RNA (mRNA) array. RESULTS: HER4 expression was up-regulated in tumor spheres compared with the expression in monolayer culture. With HER4 knockdown, NBL cells became less resistant to anoikis and serum starvation. Moreover, HER4 knockdown increased the chemosensitivity of NBL cells to cisplatin, doxorubicin, etoposide, and activated ifosfamide. In mRNA array analysis, HER4 knockdown predominately altered genes related to cell cycle regulation. In NBL spheres compared with monolayers, cell proliferation was decreased, and cyclin D expression was reduced. HER4 knockdown reversed cyclin D suppression. Overexpressed intracellular HER4 slowed the cell cycle and induced chemoresistance. CONCLUSIONS: The current results indicated that HER4 protects NBL cells from multiple exogenous apoptotic stimuli, including anoikis, nutrient deficiency, and cytotoxic chemotherapy. The intracellular fragment of HER4 was sufficient to confer this phenotype. HER4 functions as a cell cycle suppressor, maintaining resistance to cellular stress. The current findings indicate that HER4 overexpression may be associated with refractory disease, and HER4 may be an important therapeutic target.