Topical GZ21T Inhibits the Growth of Actinic Keratoses in a UVB-Induced Model of Skin Carcinogenesis.

Actinic keratoses (AKs) are premalignant intraepidermal neoplasms that occur as a result of cumulative sun damage. AKs commonly relapse, and up to 16% undergo malignant transformation into cutaneous squamous cell carcinoma. There is a need for novel therapies that reduce the quantity and surface area of AKs as well as prevent malignant transformation to cutaneous squamous cell carcinomas. We recently showed that GZ17-6.02, an anticancer agent composed of curcumin, haramine, and isovanillin, inhibited the growth of H297.T cells. This study evaluated the efficacy of a topical formulation of GZ17-6.02, known as GZ21T, in a murine model of AK generated by exposing SKH1 mice to UVR. Treatment of mice with topical GZ21T inhibited the growth of AKs by decreasing both lesion count (P = 0.012) and surface area occupied by tumor (P = 0.002). GZ21T also suppressed the progression of AKs to cutaneous squamous cell carcinoma by decreasing the count (P = 0.047) and surface area (P = 0.049) of lesions more likely to represent cutaneous squamous cell carcinoma. RNA sequencing and proteomic analyses revealed that GZ21T suppressed several pathways, including MAPK (P = 0.025), phosphoinositide 3-kinase-protein kinase B (P = 0.04), HIF-1α (P = 0.016), Wnt (P = 0.025), insulin (P = 0.018), and ERBB (P = 0.016) signaling. GZ21T also upregulated the autophagy-promoting protein AMPK while suppressing proteins such as PD-L1, glutaminase, pAkt1 S473, and eEF2K.

Construction of a Secondary Enclosure for UVB Irradiation of Mice.

UV irradiation is commonly used in murine models of skin cancers. Despite the popularity of using UVB rays to model photocarcinogenesis in animals, there is a lack of standardization in the secondary enclosures used to administer radiation. An appraisal of the literature also shows a general lack of details regarding the materials and procedures utilized in the fabrication of such enclosures. We present in this study a detailed overview of the construction of a UVB exposure chamber that successfully induces lesions in hairless mice. A standardized protocol for producing a UVB enclosure may reduce methodological variation in future studies seeking to investigate photocarcinogenesis in animals.

GZ17-6.02 Inhibits the Growth of EGFRvIII+ Glioblastoma.

Epidermal Growth Factor Receptor (EGFR) is amplified in over 50% of glioblastomas and promotes tumor formation and progression. However, attempts to treat glioblastoma with EGFR tyrosine kinase inhibitors have been unsuccessful thus far. The current standard of care is especially poor in patients with a constitutively active form of EGFR, EGFRvIII, which is associated with shorter survival time. This study examined the effect of GZ17-6.02, a novel anti-cancer agent undergoing phase 1 studies, on two EGFRvIII+ glioblastoma stem cells: D10-0171 and D317. In vitro analyses showed that GZ17-6.02 inhibited the growth of both D10-0171 and D317 cells with IC50 values of 24.84 and 28.28 µg/mL respectively. RNA sequencing and reverse phase protein array analyses revealed that GZ17-6.02 downregulates pathways primarily related to steroid synthesis and cell cycle progression. Interestingly, G17-6.02's mechanism of action involves the downregulation of the recently identified glioblastoma super-enhancer genes WSCD1, EVOL2, and KLHDC8A. Finally, a subcutaneous xenograft model showed that GZ17-6.02 inhibits glioblastoma growth in vivo. We conclude that GZ17-6.02 is a promising combination drug effective at inhibiting the growth of a subset of glioblastomas and our data warrants further preclinical studies utilizing xenograft models to identify patients that may respond to this drug.

Spontaneous Emulsions: Adjusting Spontaneity and Phase Behavior by Hydrophilic-Lipophilic Difference-Guided Surfactant, Salt, and Oil Selection.

Spontaneous emulsion behavior has been difficult to predict and could be influenced by many variables including salinity, temperature, and chemical composition of the oil and surfactant. In this work, the hydrophilic-lipophilic difference (HLD) framework was used to predict the formation of spontaneous emulsions using a mixture of Span-80 and SLES surfactants. The spontaneity and emulsion behavior of different systems were modeled by estimating the HLDmix. The influence of surfactant ratio, salinity, and oil type was investigated. Spontaneous emulsification could only be observed when the HLDmix was between -0.96 and 1.04. Within this range, a negative HLDmix resulted in a greater spontaneity to form o/w emulsion, and a w/o emulsion was more likely to form when the HLDmix was positive. When the HLDmix was close to 0 (between -0.22 and 0.56 in our systems), emulsions were formed in both the oil and aqueous phases with high spontaneity. A combined effect of ultralow interfacial tension, Span-80 micelle swelling, and interfacial turbulence due to Marangoni effects is likely the main mechanism of the spontaneous emulsification observed in this study. A synergistic reduction in interfacial tension was observed between Span-80 and SLES (<1 mN/m). When the HLD of the system was close to 0, a bicontinuous emulsion phase was formed at the oil-water interface. The bicontinuous emulsion broke-up over time due to the ultralow interfacial tension and interfacial turbulence, forming dispersed oil and water droplets. Results from this work provide a practical method to suggest what surfactant composition, salinity, and oil type could promote (or eliminate) the conditions favorable for spontaneous emulsification.

Predicting Spontaneous Emulsification in Saltwater Environments Using the HLD Model.

Spontaneous emulsification of toluene with nonylphenol polyethoxylate (NPE) and sodium dodecylbenzenesulfonate (SDBS) surfactants in saltwater environments was studied. NaCl promoted the spontaneous emulsification of an otherwise non-spontaneous SDBS-toluene system. Dynamic light scattering and turbidity indicated that spontaneity increased with NaCl concentration. The mechanism of spontaneous emulsification was dependent on surfactant type; NPE emulsified via micelle swelling, and SDBS emulsified via nucleation and growth. Hydrophilic lipophilic difference (HLD) calculations were used to model spontaneous emulsification and spontaneity. As HLD approached zero, conditions became more favorable for spontaneous emulsification. Between HLD values of -2.4 and -2.05, samples transitioned from non-spontaneous to spontaneous. This study aids in predicting spontaneous emulsion formation in saltwater environments for applications in nanoemulsion formation and wastewater remediation.

Diffusion-Controlled Spontaneous Emulsification of Water-Soluble Oils via Micelle Swelling.

Spontaneous emulsification of toluene, xylenes, cyclohexane, and mineral oil in a nonionic nonylphenol polyethoxylate surfactant solution was investigated by visual observations coupled with dynamic light scatting measurements and interfacial tensiometry. For water-soluble oils, nanoscale emulsions formed spontaneously by diffusion of oil molecules into the aqueous surfactant solutions and subsequent swelling of surfactant micelles with oil. Micelle swelling rates were quantified to assess system spontaneity, revealing that oil solubility in water was directly correlated to the spontaneity of the emulsion (toluene > xylenes > cyclohexane). When experiments were intentionally designed to create surfactant concentration gradients, Marangoni flows were found to enhance spontaneity. Despite their spontaneous formation, emulsion stability was limited over the course of 40 days by Ostwald ripening followed by creaming and evaporation. These results provide insights on the likelihood of nanoemulsion formation and persistence in oily wastewater as the components in this study are present in many wastewater systems.

Poor agreement of prostate specific antigen doubling times calculated using ultrasensitive versus standard prostate specific antigen values: important impact on risk assessment.

PURPOSE: In men with biochemical recurrence after radical prostatectomy, a rapid prostate specific antigen doubling time is associated with adverse outcomes, and is often used to guide the type and timing of salvage therapy. It is unknown whether prostate specific antigen doubling time calculated in the ultrasensitive range (prostate specific antigen less than 0.2 ng/ml) accurately reflects measures performed in the traditional range (prostate specific antigen greater than 0.2 ng/ml). MATERIALS AND METHODS: We studied 394 men in a national disease registry of men with prostate cancer (CaPSURE™) who underwent radical prostatectomy, experienced biochemical failure, and had prostate specific antigen doubling time assessed using ultrasensitive and traditional prostate specific antigen values. Agreement between these measurements was assessed using Cohen's kappa score. RESULTS: Median ultrasensitive prostate specific antigen doubling time was 11.9 months (IQR 6-29) and median traditional prostate specific antigen doubling time was 240 months (IQR 18-240). Agreement between ultrasensitive and traditional prostate specific antigen doubling time was poor, with a weighted Cohen's kappa score of 0.04 (95% CI -0.02-0.10). Using a dichotomous prostate specific antigen doubling time cutoff of 9 months, there was a statistically significant difference between ultrasensitive and standard prostate specific antigen doubling time (exact McNemar p <0.01). Ultrasensitive prostate specific antigen doubling time was more or less rapid than traditional prostate specific antigen doubling time by more than 15 months in 244 (62%) and 35 (9%) patients, respectively. CONCLUSIONS: Agreement between prostate specific antigen doubling time calculated using ultrasensitive vs traditional prostate specific antigen values is poor. Ultrasensitive prostate specific antigen doubling time is often significantly more rapid than traditional prostate specific antigen doubling time, potentially overestimating the risk of clinical recurrence. Until the significance of ultrasensitive prostate specific antigen doubling time is better characterized, the decision to proceed with salvage therapy should not be based on prostate specific antigen doubling time calculated using ultrasensitive prostate specific antigen values.

Incidence of bladder neck contracture after robot-assisted laparoscopic and open radical prostatectomy.

OBJECTIVE: To evaluate the incidence and risk factors for bladder neck contracture (BNC) in men treated with robot-assisted laparoscopic radical prostatectomy (RALP) and open radical prostatectomy (ORP), as BNC is a well-described complication of ORP and may be partially attributable to technique. PATIENTS AND METHODS: The University of California San Francisco Urologic Oncology Database was queried for patients undergoing RALP or ORP from 2002 to 2008. Patient demographics, prostate cancer-specific information, surgical data, and follow-up were collected. For each surgical approach, multivariate Cox proportional hazards regression was performed to evaluate associations of demographics and clinical characteristics with BNC. Time to BNC after RP was evaluated using life table and Kaplan-Meier methods. RESULTS: From 2002 to 2008, 988 patients underwent RP as primary treatment and had at least 12 months of follow-up. Of these men, 695 underwent ORP and 293 underwent RALP. The mean (sd) age was 59.3 (6.80) years and 91% of men were Caucasian. D'Amico risk groups at diagnosis were low (38%), intermediate (38%), and high (24%). The BNC incidence was 2.2% (22 cases) overall, 1.4% (four) for RALP, and 2.6% (18) for ORP (P= 0.12). Patients with BNC were diagnosed a median (range) of 4.7 (1-15) months after surgery. At 18 months after surgery, the BNC-free rate was 97% for ORP and 99% for RALP (log-rank P= 0.13). The most common presenting complaint was slow stream, followed by urinary retention. In Cox proportional hazards regression analysis, earlier year of surgery, older age at diagnosis and higher PSA level at diagnosis were significantly associated with BNC among ORP patients. In the RALP group, none of the covariates were associated with BNC. CONCLUSIONS: The overall incidence of BNC was low in both RALP and ORP groups. Technical factors such as enhanced magnification and a running bladder anastomosis may explain the lower BNC incidence in the RALP group.

Genetic dissection of the oncogenic mTOR pathway reveals druggable addiction to translational control via 4EBP-eIF4E.

We genetically dissect the contribution of the most prominent downstream translational components of mTOR signaling toward Akt-driven lymphomagenesis. While phosphorylation of rpS6 is dispensable for cancer formation, 4EBP-eIF4E exerts significant control over cap-dependent translation, cell growth, cancer initiation, and progression. This effect is mediated at least in part through 4EBP-dependent control of Mcl-1 expression, a key antiapoptotic protein. By using an active site inhibitor of mTOR, PP242, we show a marked therapeutic response in rapamycin-resistant tumors. The therapeutic benefit of PP242 is mediated through inhibition of mTORC1-dependent 4EBP-eIF4E hyperactivation. Thus, the 4EBP-eIF4E axis downstream of mTOR is a druggable mediator of translational control and Akt-mediated tumorigenesis that has important implications for the treatment of human cancers.

Penile sonographic and clinical characteristics in men with Peyronie's disease.

INTRODUCTION: Ultrasonography of the penis is readily available to the urologist and gives good anatomic detail of soft tissue structures. It has not been widely utilized in the assessment of Peyronie's disease (PD). AIMS: To describe the sonographic characteristics of the penis in PD and the relationship between clinical and sonographic features. METHODS: This cross-sectional study enrolled patients from a single clinical practice. A PD-specific questionnaire was administered and sonographic evaluations were performed. MAIN OUTCOME MEASURES: Sonographic characteristics of men with PD. RESULTS: Tunical thickening, calcifications, septal fibrosis, and intracavernosal fibrosis, were observed at initial clinical evaluation in 50%, 31%, 20%, and 15% of men, respectively. Men aged 40-49 (OR 2.4, P = 0.02) and men aged 50-59 (OR 2.4, P = 0.004) were more likely to have sub-tunical calcifications relative to men under age 40. Men with septal fibrosis had fewer chronic medical conditions such as diabetes (OR 0.3, P = 0.04), hypertension (OR 0.5, P = 0.03), and coronary artery disease (OR 0.2, P = 0.05), and presented within 1 year of disease onset (OR 2.1, P = 0.001). Men with septal fibrosis were less likely to have lost penile length (OR 0.5, P = 0.04) and more likely to be able to have intercourse (OR 1.9, P = 0.05). Men with intracavernosal fibrosis were less likely to have penile pain (OR 0.5, P = 0.05), but more likely to have penetration difficulty during intercourse (OR 1.9, P = 0.008), an additional penile deformity (OR 1.8, P = 0.02), or rapid onset of disease (OR 1.7, P = 0.04). Tunical thickening was associated with a decreased ability to have intercourse (OR 2.3, P < 0.001). CONCLUSION: PD is a clinically and sonographically heterogeneous condition. Sonography is a safe, low-cost, and rapid means of objectively characterizing lesions in this condition. This may help track the evolution of the condition in individual patients and in the future may be useful for tailoring treatment strategies.

Optical coherence tomography: a novel modality for scrotal imaging.

BACKGROUND: For patients with nonobstructive azoospermia, sperm retrieval rates remain modest. We describe the use of optical coherence tomography to improve retrieval rates and to decrease tissue destruction. METHODS: Four patients underwent diagnostic testicular biopsy and imaging with the Niris optical coherence tomography device. We performed a descriptive comparison between optical coherence tomographic images and conventional histology. RESULTS: The measured seminiferous tubule diameter differed by 16 mum between comparative imaging from optical coherence tomography and conventional histology using hematoxylin and eosin staining. CONCLUSION: We illustrate the usefulness of optical coherence tomography in the setting of testicular biopsy and the management of nonobstructive azoospermia.