Study of secondary organic aerosol formation and aging using ambient air in an oxidation flow reactor during high pollution events over Delhi.

Secondary aerosols constitute a significant fraction of atmospheric aerosols, yet our understanding of their formation mechanism and fate is very limited. In this work, the secondary organic aerosol (SOA) formation and aging of ambient air of Delhi are studied using a potential aerosol mass (PAM) reactor, an oxidation flow reactor (OFR), coupled with aerosol chemical speciation monitor (ACSM), proton transfer reaction time of flight mass spectrometer (PTR-ToF-MS), and scanning mobility particle sizer with counter (SMPS + C). The setup mimics atmospheric aging of up to several days with the generation of OH radicals. Variations in primary volatile organic compounds (VOCs) and oxygenated volatile organic compounds (OVOCs) as a function of photochemical age were investigated. Primary VOCs such as benzene, toluene, xylene, trimethyl benzene, etc. decrease and OVOCs like formic acid, formaldehyde, acetone, ethanol, etc. increase substantially upon oxidation in OFR. The highest organic aerosol (OA) enhancement was observed for the 4.2 equivalent photochemical days of aging i.e., 1.84 times the ambient concentration, and net OA loss was observed at very high OH exposure, typically after 8.4 days of photochemical aging due to heterogeneous oxidation followed by fragmentation/evaporation. In ambient air, OA enhancement is highest during nighttime due to the high concentrations of precursor VOCs in the atmosphere. SMPS + C results demonstrated substantial new particle formation upon aging and decrement in preexisting aerosol mass. This is the first experimental study conducting an in-situ evaluation of potential SOA mass generated from the ambient aerosols in India.

Spatiotemporal variability and health risk assessment of PM2.5 and NO2 over the Indo-Gangetic Plain: A three years long study (2019-21).

Studying the spatiotemporal variability of pollutants is necessary to identify the pollution hotspots with high health risk and enable the agencies to implement pollution abatement strategies in a targeted manner. Present study reports the spatio-temporal variability and health risk assessment (HRA) of PM2.5 (Particulate matter with aerodynamic diameter <2.5µm) and NO2 over IGP from 2019-2021. The HRA is expressed as passively smoked cigarettes (PSC) for four different health outcomes i.e., low birth weight (LBW), percentage decreased lung function (DLF) in school aged children, lung cancer (LC), and cardiovascular mortality (CM). The findings confirm very high PM2.5 and NO2 mass concentrations and high health risk over middle IGP and Delhi as compared to upper and lower IGP. Within Delhi, north Delhi region is the most polluted and at highest risk as compared to central and south Delhi. The health risk associated with PM2.5 over IGP is highest for DLF, equivalent to 21.63 PSCs daily, followed by CM (11.69), LBW (8.27) and LC (6.94). For NO2, the health risk is highest for DLF (3.09 PSCs) and CM (2.95), followed by LC (1.47) and LBW (1.04). PM2.5 and NO2 concentrations, along with the associated health risks, are highest during the post-monsoon and winter seasons and lowest during the monsoon season.

Activating point mutations in the MET kinase domain represent a unique molecular subset of lung cancer and other malignancies targetable with MET inhibitors.

Activating point mutations in the MET tyrosine kinase domain (TKD) are oncogenic in a subset of papillary renal cell carcinomas (PRCC). Here, using comprehensive genomic profiling among >600,000 patients, we identify activating MET TKD point mutations as putative oncogenic driver across diverse cancers, with a frequency of ~0.5%. The most common mutations in the MET TKD defined as oncogenic or likely oncogenic according to OncoKB resulted in amino acid substitutions at positions H1094, L1195, F1200, D1228, Y1230, M1250, and others. Preclinical modeling of these alterations confirmed their oncogenic potential, and also demonstrated differential patterns of sensitivity to type I and type II MET inhibitors. Two patients with metastatic lung adenocarcinoma harboring MET TKD mutations (H1094Y, F1200I) and no other known oncogenic drivers achieved confirmed partial responses to a type I MET inhibitor. Activating MET TKD mutations occur in multiple malignancies and may confer clinical sensitivity to currently available MET inhibitors.

A role for ATF2 in regulating MITF and melanoma development.

The transcription factor ATF2 has been shown to attenuate melanoma susceptibility to apoptosis and to promote its ability to form tumors in xenograft models. To directly assess ATF2's role in melanoma development, we crossed a mouse melanoma model (Nras(Q61K)::Ink4a⁻/⁻) with mice expressing a transcriptionally inactive form of ATF2 in melanocytes. In contrast to 7/21 of the Nras(Q61K)::Ink4a⁻/⁻ mice, only 1/21 mice expressing mutant ATF2 in melanocytes developed melanoma. Gene expression profiling identified higher MITF expression in primary melanocytes expressing transcriptionally inactive ATF2. MITF downregulation by ATF2 was confirmed in the skin of Atf2⁻/⁻ mice, in primary human melanocytes, and in 50% of human melanoma cell lines. Inhibition of MITF transcription by MITF was shown to be mediated by ATF2-JunB-dependent suppression of SOX10 transcription. Remarkably, oncogenic BRAF (V600E)-dependent focus formation of melanocytes on soft agar was inhibited by ATF2 knockdown and partially rescued upon shMITF co-expression. On melanoma tissue microarrays, a high nuclear ATF2 to MITF ratio in primary specimens was associated with metastatic disease and poor prognosis. Our findings establish the importance of transcriptionally active ATF2 in melanoma development through fine-tuning of MITF expression.

Source apportionment, health risk assessment, and trajectory analysis of black carbon and light absorption properties of black and brown carbon in Delhi, India.

Black Carbon (BC) is an important atmospheric pollutant, well recognized for adverse health and climatic effects. The present work discusses the monthly and seasonal variations of BC sources, health risks, and light absorption properties. The measurement was done from January to December 2021 using a seven wavelength aethalometer. Annual average BC concentration during the study period was 12.2 ± 8.8 µg/m3 (ranged from 1.9 - 52.2 µg/m3). Results represent highest BC concentration during winter (W), followed by post-monsoon (P-M), summer (S), and monsoon (M) seasons where the fossil fuel (FF) combustion is the major source during W, S, and M seasons and biomass burning (BB) during the P-M season. The health risk assessment revealed that individuals in Delhi are exposed to BC levels equivalent to inhaling the smoke from 36 passively smoked cigarettes (PSC) everyday. The risk is highest during W reaching upto 71 PSC and minimum during M i.e., 9 PSC. The light absorption properties were calculated for BC (AbsBC) and Brown carbon (AbsBrC). AbsBC and varied from 229-89 Mm-1 between 370-950 nm and AbsBrC varied from 87-12 Mm-1 between 370-660 nm. AbsBC contributed substantially to total absorption at all wavelengths, while AbsBrC contribution is quite significant in the UV region only. Trajectory analysis confirmed significant influence of regional sources (e.g., biomass-burning aerosols from northwest and east direction) on air quality, health risks, and light absorption properties of BC over Delhi especially during the P-M season. The BB events of Punjab, Haryana, Uttar Pradesh, and eastern Pakistan seems to have significant influence on Delhi's air quality predominantly during P-M season.

Key Aggregation Cryptosystem and Double Encryption Method for Cloud-Based Intelligent Machine Learning Techniques-Based Health Monitoring Systems.

Cloud technology is a business strategy that aims to provide the necessary material to customers depending on their needs. Individuals and cloud businesses alike have embraced the cloud storage service, which has become the most widely used service. The industries outsource their data to cloud storage space to relieve themselves of the load of dealing with redundant data contents. This must be protected to prevent the theft of personal belongings, and privacy must be improved as well. Different research projects have been suggested to ensure the safe management of the information included within the data content. The security of current research projects, on the contrary, still needs improvement. As a result, this method has been suggested to address the security concerns associated with cloud computing. The primary goal of this study effort is to offer a safe environment for cloud users while also increasing the profit of cloud resource providers by managing and securely delivering data contents to the cloud users. The bulk of sectors, including business, finance, military, and healthcare industry, do not store data in cloud-based storage systems. This technique is used to attract these kinds of customers. Increasing public acceptance, medical researchers are drawn to cloud computing because it allows them to store their study material in a centralized location and distribute and access it in a more flexible manner. They were collected from numerous individuals who were being evaluated for medical care at the time. Scalable and enhanced key aggregate cryptosystem is a protected data protection method that provides highly effective security in the healthcare industry. When parties interested in a dispute disagree on the outflow of sensitive information, this technique manages the disputes and ensures the data security deployment of a cloud-based intelligent health monitoring system for the parties involved. The encrypted data structure of medical and healthcare prescriptions is recorded as they move through the hands of patients and healthcare facilities, according to the technique recommended. The double encryption approach is used in order to raise the overall degree of security. An encryption class is created by referring to the Ciphertext ID during the encryption procedure. The keyholder is a master secret key that facilitates in the recovery of the secret keys of various monsters and creatures by acting as a conduit between them. It is transferred and stored as a single aggregate for the benefit of the patient or customer in order to make decryption more convenient and efficient. A safe connection between cloud-based intelligent health monitoring systems and healthcare organizations and their patients may be established via the use of a key aggregation cryptosystem and a double encryption approach, according to the researchers. Because of this, when compared to earlier techniques, the findings reveal that the research methodology provides high levels of security in terms of confidentiality and integrity, in addition to excellent scalability.

Machine Learning Techniques for Human Age and Gender Identification Based on Teeth X-Ray Images.

The use of digital medical images is increasing with advanced computational power that has immensely contributed to developing more sophisticated machine learning techniques. Determination of age and gender of individuals was manually performed by forensic experts by their professional skills, which may take a few days to generate results. A fully automated system was developed that identifies the gender of humans and age based on digital images of teeth. Since teeth are a strong and unique part of the human body that exhibits least subject to risk in natural structure and remains unchanged for a longer duration, the process of identification of gender- and age-related information from human beings is systematically carried out by analyzing OPG (orthopantomogram) images. A total of 1142 digital X-ray images of teeth were obtained from dental colleges from the population of the middle-east part of Karnataka state in India. 80% of the digital images were considered for training purposes, and the remaining 20% of teeth images were for the testing cases. The proposed gender and age determination system finds its application widely in the forensic field to predict results quickly and accurately. The prediction system was carried out using Multiclass SVM (MSVM) classifier algorithm for age estimation and LIBSVM classifier for gender prediction, and 96% of accuracy was achieved from the system.

Impact of Chemical Properties of Human Respiratory Droplets and Aerosol Particles on Airborne Viruses' Viability and Indoor Transmission.

The airborne transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as a potential pandemic challenge, especially in poorly ventilated indoor environments, such as certain hospitals, schools, public buildings, and transports. The impacts of meteorological parameters (temperature and humidity) and physical property (droplet size) on the airborne transmission of coronavirus in indoor settings have been previously investigated. However, the impacts of chemical properties of viral droplets and aerosol particles (i.e., chemical composition and acidity (pH)) on viability and indoor transmission of coronavirus remain largely unknown. Recent studies suggest high organic content (proteins) in viral droplets and aerosol particles supports prolonged survival of the virus by forming a glassy gel-type structure that restricts the virus inactivation process under low relative humidity (RH). In addition, the virus survival was found at neutral pH, and inactivation was observed to be best at low (<5) and high pH (>10) values (enveloped bacteriophage Phi6). Due to limited available information, this article illustrates an urgent need to research the impact of chemical properties of exhaled viral particles on virus viability. This will improve our fundamental understanding of indoor viral airborne transmission mechanisms.

Cerebral hemodynamics in children with sickle cell disease in India: An observational cohort study.

India has the second highest number of cases of sickle cell disease (SCD) and affects the most socioeconomically disadvantaged communities living in a horizontal belt from Gujarat to Odisha state. Despite high prevalence, information about cerebral hemodynamics among children with SCD in India remains scarcely described. We performed transcranial Doppler (TCD) to assess cerebral hemodynamics among Indian children with SCD and evaluated their association with clinical and hematological parameters. Children aged 3-18years, diagnosed with SCD living in Raipur in Chhattisgarh and Ahmedabad in Gujarat state were recruited. TCD was performed to obtain flow velocities from middle cerebral (MCA), intracranial internal carotid (ICA) and basilar artery. Associations were evaluated between timed-average-mean-maximum velocities (TAMMV) and various clinical and hematological parameters. Our prospective study included 62 consecutive children with known SCD. Mean ± SD age of the study population was 9.8 ± 3.9 years and 31 (50%) were male. Mean ± SD hemoglobin was 8.64 ± 1.34 Gm/dL while the mean HbSS ± SD was 70.25 ± 15.27%. While 6 (9.6%) children had suffered from stroke during previous 2 years, 7 (11%) demonstrated abnormal TAMMV. Higher HbSS level along with history of iron chelation therapy, blood transfusion and/or stroke showed a trend towards having higher TAMMV. Stroke and cerebral hemodynamic alterations are common among Indian children with SCD. Larger studies with detailed neuroimaging and genetic evaluations are needed for better understanding, characterization, risk stratification as well as optimization of the timing of blood transfusion to reduce physical disabilities among Indian children with SCD.

Variations in Black Carbon concentration and sources during COVID-19 lockdown in Delhi.

A nationwide lockdown was imposed in India due to COVID-19 pandemic in five phases from 25th March to May 31, 2020. The lockdown restricted major anthropogenic activities, primarily vehicular and industrial, thereby reducing the particulate matter concentration. This work investigates the variation in Black Carbon (BC) concentration and its sources (primarily Fossil Fuel (ff) burning and Biomass Burning (bb)) over Delhi from 18th February to July 31, 2020, covering one month of pre-lockdown phase, all the lockdown phases, and two months of successive lockdown relaxations. The daily average BC concentration varied from 0.22 to 16.92 µg/m3, with a mean value of 3.62 ± 2.93 µg/m3. During Pre-Lockdown (PL, 18th Feb-24th March 2020), Lockdown-1 (L1, 25th March-14th April 2020), Lockdown-2 (L2, 15th April-3rd May 2020), Lockdown-3 (L3, 4th-17th May 2020), Lockdown-4 (L4, 18th-31st May 2020), Unlock-1 (UN1, June 2020), and Unlock-2 (UN2, July 2020) the average BC concentrations were 7.93, 1.73, 2.59, 3.76, 3.26, 2.07, and 2.70 µg/m3, respectively. During the lockdown and unlock phases, BC decreased up to 78% compared to the PL period. The BC source apportionment studies show that fossil fuel burning was the dominant BC source during the entire sampling period. From L1 to UN2 an increasing trend in BCff contribution was observed (except L3) due to the successive relaxations given to anthropogenic activities. BCff contribution dipped briefly during L3 due to the intensive crop residue burning events in neighboring states. CWT analysis showed that local emission sources were the dominant contributors to BC concentration over Delhi.

Variation in chemical composition and sources of PM2.5 during the COVID-19 lockdown in Delhi.

The Government of India (GOI) announced a nationwide lockdown starting 25th March 2020 to contain the spread of COVID-19, leading to an unprecedented decline in anthropogenic activities and, in turn, improvements in ambient air quality. This is the first study to focus on highly time-resolved chemical speciation and source apportionment of PM2.5 to assess the impact of the lockdown and subsequent relaxations on the sources of ambient PM2.5 in Delhi, India. The elemental, organic, and black carbon fractions of PM2.5 were measured at the IIT Delhi campus from February 2020 to May 2020. We report source apportionment results using positive matrix factorization (PMF) of organic and elemental fractions of PM2.5 during the different phases of the lockdown. The resolved sources such as vehicular emissions, domestic coal combustion, and semi-volatile oxygenated organic aerosol (SVOOA) were found to decrease by 96%, 95%, and 86%, respectively, during lockdown phase-1 as compared to pre-lockdown. An unforeseen rise in O3 concentrations with declining NOx levels was observed, similar to other parts of the globe, leading to the low-volatility oxygenated organic aerosols (LVOOA) increasing to almost double the pre-lockdown concentrations during the last phase of the lockdown. The effect of the lockdown was found to be less pronounced on other resolved sources like secondary chloride, power plants, dust-related, hydrocarbon-like organic aerosols (HOA), and biomass burning related emissions, which were also swayed by the changing meteorological conditions during the four lockdown phases. The results presented in this study provide a basis for future emission control strategies, quantifying the extent to which constraining certain anthropogenic activities can ameliorate the ambient air. These results have direct relevance to not only Delhi but the entire Indo-Gangetic plain (IGP), citing similar geographical and meteorological conditions common to the region along with overlapping regional emission sources. SUMMARY OF MAIN FINDINGS: We identify sources like vehicular emissions, domestic coal combustion, and semi-volatile oxygenated organic aerosol (SVOOA) to be severely impacted by the lockdown, whereas ozone levels and, in turn, low-volatility oxygenated organic aerosols (LVOOA) rise by more than 95% compared to the pre-lockdown concentrations during the last phase of the lockdown. However, other sources resolved in this study, like secondary chloride, power plants, dust-related, hydrocarbon-like organic aerosols (HOA), and biomass burning related emissions, were mainly driven by the changes in the meteorological conditions rather than the lockdown.

Zotatifin, an eIF4A-Selective Inhibitor, Blocks Tumor Growth in Receptor Tyrosine Kinase Driven Tumors.

Oncoprotein expression is controlled at the level of mRNA translation and is regulated by the eukaryotic translation initiation factor 4F (eIF4F) complex. eIF4A, a component of eIF4F, catalyzes the unwinding of secondary structure in the 5'-untranslated region (5'-UTR) of mRNA to facilitate ribosome scanning and translation initiation. Zotatifin (eFT226) is a selective eIF4A inhibitor that increases the affinity between eIF4A and specific polypurine sequence motifs and has been reported to inhibit translation of driver oncogenes in models of lymphoma. Here we report the identification of zotatifin binding motifs in the 5'-UTRs of HER2 and FGFR1/2 Receptor Tyrosine Kinases (RTKs). Dysregulation of HER2 or FGFR1/2 in human cancers leads to activation of the PI3K/AKT and RAS/ERK signaling pathways, thus enhancing eIF4A activity and promoting the translation of select oncogenes that are required for tumor cell growth and survival. In solid tumor models driven by alterations in HER2 or FGFR1/2, downregulation of oncoprotein expression by zotatifin induces sustained pathway-dependent anti-tumor activity resulting in potent inhibition of cell proliferation, induction of apoptosis, and significant in vivo tumor growth inhibition or regression. Sensitivity of RTK-driven tumor models to zotatifin correlated with high basal levels of mTOR activity and elevated translational capacity highlighting the unique circuitry generated by the RTK-driven signaling pathway. This dependency identifies the potential for rational combination strategies aimed at vertical inhibition of the PI3K/AKT/eIF4F pathway. Combination of zotatifin with PI3K or AKT inhibitors was beneficial across RTK-driven cancer models by blocking RTK-driven resistance mechanisms demonstrating the clinical potential of these combination strategies.

Influence of chemical aging on physico-chemical properties of mineral dust particles: A case study of 2016 dust storms over Delhi.

The physico-chemical properties of dust particles collected During Dust Storm (DDS) and After Dust Storm (ADS) events were studied using Scanning Electron Microscope coupled with Energy Dispersive X-ray Spectroscopy (SEM-EDS), X-ray Fluorescence Spectroscopy (XRF) and X-ray Photoelectron Spectroscopy (XPS). Morphological and compositional change in dust particles were observed as they react with the anthropogenic pollutants present in the urban environment. The calcite rich particles were observed to transform into calcium chloride, calcium nitrate, and calcium sulfate on reacting with the chlorides, nitrates, and sulfates present in the urban atmosphere. The frequency distributions of Aspect Ratio (AR) for the DDS and ADS particles were observed to be bimodal (mode peaks at 1.2 and 1.5) and monomodal (mode peak at 1.1), respectively. The highly irregular shaped solid dust particles were observed to transform into nearly spherical semisolid particles in the urban environment. XPS analysis confirms the high concentration of oxides, nitrates, and chlorides at the surface of ADS samples which show the signatures of mineral dust particles aging. Species with a high value of imaginary part of refractive index (like Cr metal, Fe metal, Cr2O3, FeO, Fe2O3) were observed at the surface of dust particles. At 550 nm wavelength, the light-absorbing potential of the observed species along with black carbon (BC) was found to vary in the order; Cr metal > Fe metal > Cr2O3> FeO > BC > Fe2O3> FeOOH. The presence of the aforementioned species on the surface of ADS particles will tremendously affect the particle optical and radiative properties compared to that of DDS particles. The present work could reduce the uncertainty in the radiation budget estimations of mineral dust and assessment of their climatic impacts over Delhi.

Structure-based Design of Pyridone-Aminal eFT508 Targeting Dysregulated Translation by Selective Mitogen-activated Protein Kinase Interacting Kinases 1 and 2 (MNK1/2) Inhibition.

Dysregulated translation of mRNA plays a major role in tumorigenesis. Mitogen-activated protein kinase interacting kinases (MNK)1/2 are key regulators of mRNA translation integrating signals from oncogenic and immune signaling pathways through phosphorylation of eIF4E and other mRNA binding proteins. Modulation of these key effector proteins regulates mRNA, which controls tumor/stromal cell signaling. Compound 23 (eFT508), an exquisitely selective, potent dual MNK1/2 inhibitor, was designed to assess the potential for control of oncogene signaling at the level of mRNA translation. The crystal structure-guided design leverages stereoelectronic interactions unique to MNK culminating in a novel pyridone-aminal structure described for the first time in the kinase literature. Compound 23 has potent in vivo antitumor activity in models of diffuse large cell B-cell lymphoma and solid tumors, suggesting that controlling dysregulated translation has real therapeutic potential. Compound 23 is currently being evaluated in Phase 2 clinical trials in solid tumors and lymphoma. Compound 23 is the first highly selective dual MNK inhibitor targeting dysregulated translation being assessed clinically.

Genetic alterations in signaling pathways in melanoma.

Alterations in the RAS signaling cascade are almost uniformly present in melanoma. RAS itself is only infrequently mutated in melanoma although downstream of RAS lie BRAF on the mitogen-activated protein kinase pathway and PTEN on the protein kinase B/Akt pathway. These genes are often altered in melanomas; indeed, the most frequent target of mutation in melanomas is BRAF, which is mutated in approximately 60% to 70% of superficial spreading melanomas. These mutations occur in a background that is not normal, with the CDKN2A locus also typically being mutated. We review herein the data that suggest that the distribution of the signaling mutations is important. In general, melanomas carry a mutated NRAS, a mutated BRAF, or concurrent BRAF and PTEN mutations. These data support the hypothesis that the biochemical functions of RAS are portioned by mutations in the pathways lying downstream. Moreover, these mutations have no apparent relationship to the patterns of alteration of CDKN2A and its downstream effectors. Thus, the data also suggest that successful exploitation of mutations in melanoma will be dependent on understanding not only mutations and their frequency but their genetic context as well.

Examination of mutations in BRAF, NRAS, and PTEN in primary cutaneous melanoma.

Frequent somatic mutation of v-raf murine sarcoma viral oncogene homolog B (BRAF), a downstream effector of the rat sarcoma oncogene (RAS) signaling pathway, is described in melanoma and other tumors. Our analysis of melanoma cell lines suggests that activating mutations in BRAF can occur simultaneously with inactivation of phosphatase and tensin homolog (PTEN), but neuroblastoma RAS (NRAS) mutations are not coincident. We determined the concurrent prevalence of mutations in BRAF and NRAS, and alteration of PTEN expression in 69 primary cutaneous melanomas. BRAF mutations were seen in 57% of cases. NRAS was mutated in 17% of samples, exclusively in exon 2. Two cases showed concurrent BRAF and NRAS mutations. Using immunohistochemistry, PTEN protein expression was lost or greatly reduced in 19% of tumors. Seven tumors with reduced PTEN yielded DNA amenable to sequencing, and three also showed mutation in BRAF but none in NRAS. In all, 11 (85%) of 13 tumors showing reduced PTEN expression were greater than 3.5 mm thick, and the association of increasing Breslow thickness and loss or reduction of PTEN expression was statistically significant (P<0.0001). Mutations in NRAS were not coincident with reduced PTEN expression, and the concurrent mutation of NRAS and BRAF was rare.

PTEN signaling pathways in melanoma.

Phosphatase and tensin homolog deleted in from chromosome ten (PTEN), initially also known as mutated in multiple advanced cancers or TGF-beta-regulated and epithelia cell-enriched phosphatase, is a tumor suppressor gene that is mutated in a large fraction of human melanomas. A broad variety of human cancers carry PTEN alterations, including glioblastomas, endometrial, breast, thyroid and prostate cancers. The PTEN protein has at least two biochemical functions: it has both lipid phosphatase and protein phosphatase activity. The lipid phosphatase activity of PTEN decreases intracellular PtdIns(3,4,5)P(3) level and downstream Akt activity. Cell-cycle progression is arrested at G1/S, mediated at least partially through the upregulation of the cyclin-dependent kinase inhibitor p27. In addition, agonist-induced apoptosis is mediated by PTEN, through the upregulation of proapoptotic machinery involving caspases and BID, and the downregulation of antiapoptotic proteins such as Bcl2. The protein phosphatase activity of PTEN is apparently less central to its involvement in tumorigenesis. It is involved in the inhibition of focal adhesion formation, cell spreading and migration, as well as the inhibition of growth factor-stimulated MAPK signaling. Therefore, the combined effects of the loss of PTEN lipid and protein phosphatase activity may result in aberrant cell growth and escape from apoptosis, as well as abnormal cell spreading and migration. In melanoma, PTEN loss has been mostly observed as a late event, although a dose-dependent loss of PTEN protein and function has been implicated in early stages of tumorigenesis as well. In addition, loss of PTEN and oncogenic activation of RAS seem to occur in a reciprocal fashion, both of which could cooperate with CDKN2A loss in contribution to melanoma tumorigenesis.

Genetic interaction between NRAS and BRAF mutations and PTEN/MMAC1 inactivation in melanoma.

Extant evidence implicates growth factor signaling in the pathogenesis of many tumor types, including cutaneous melanoma. Recently, reciprocal activating mutations of NRAS and BRAF were found in benign melanocytic nevi and cutaneous melanomas. We had previously reported a similar epistatic relationship between activating NRAS mutations and inactivating PTEN/MMAC1 alterations. We thus hypothesized that BRAF and PTEN/MMAC1 mutations may cooperate to promote melanoma tumorigenesis. Overall, 40 of 47 (85%) melanoma cell lines and 11 of 16 (69%) uncultured melanoma metastases had mutations in NRAS, BRAF, or PTEN/MMAC1. NRAS was exclusively mutated in nine of 47 (19%) cell lines and two of 16 (13%) metastases, whereas BRAF was solely mutated in 28 of 47 (60%) cell lines and nine of 16 (56%) metastases. In the 12 of 15 melanoma cell lines (80%) and two of two melanoma metastases with PTEN alterations, BRAF was also mutated. These findings suggest the existence of possible cooperation between BRAF activation and PTEN loss in melanoma development.

Pancreatic ductal adenocarcinoma contains an effector and regulatory immune cell infiltrate that is altered by multimodal neoadjuvant treatment.

OBJECTIVE: The immune response to pancreatic ductal adenocarcinoma (PDA) may play a role in defining its uniquely aggressive biology; therefore, we sought to clearly define the adaptive immune infiltrate in PDA. DESIGN: We used immunohistochemistry and flow cytometry to characterize the immune infiltrate in human PDA and compared our findings to the patients' peripheral blood. RESULTS: In contrast to the myeloid cell predominant infiltrate seen in murine models, T cells comprised the majority of the hematopoietic cell component of the tumor stroma in human PDA. Most intratumoral CD8+ T cells exhibited an antigen-experienced effector memory cell phenotype and were capable of producing IFN-γ. CD4+ regulatory T cells (Treg) and IL-17 producing T helper cells were significantly more prevalent in tumor than in blood. Consistent with the association with reduced survival in previous studies, we observed higher frequencies of both myeloid cells and Treg in poorly differentiated tumors. The majority of intratumoral T cells expressed the co-inhibitory receptor programmed death-1 (PD-1), suggesting one potential mechanism through which PDA may evade antitumor immunity. Successful multimodal neoadjuvant therapy altered the immunoregulatory balance and was associated with reduced infiltration of both myeloid cells and Treg. CONCLUSION: Our data show that human PDA contains a complex mixture of inflammatory and regulatory immune cells, and that neoadjuvant therapy attenuates the infiltration of intratumoral cells associated with immunosuppression and worsened survival.

Enzymatic targeting of the stroma ablates physical barriers to treatment of pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinomas (PDAs) are characterized by a robust fibroinflammatory response. We show here that this desmoplastic reaction generates inordinately high interstitial fluid pressures (IFPs), exceeding those previously measured or theorized for solid tumors, and induces vascular collapse, while presenting substantial barriers to perfusion, diffusion, and convection of small molecule therapeutics. We identify hyaluronan, or hyaluronic acid (HA), as the primary matrix determinant of these barriers and show that systemic administration of an enzymatic agent can ablate stromal HA from autochthonous murine PDA, normalize IFP, and re-expand the microvasculature. In combination with the standard chemotherapeutic, gemcitabine, the treatment permanently remodels the tumor microenvironment and consistently achieves objective tumor responses, resulting in a near doubling of overall survival.