Chebulinic Acid: An Incipient Anticancer Agent.

BACKGROUND: Terminalia chebula (T. chebula) comprising chebulinic acid as its principle active constituent is used to cure various diseases. T. chebula and chebulinic acid are used as antimicrobial, antioxidant, antidiabetic, anti-inflammatory, hepatoprotective, antimutagenic, radioprotective, cardioprotective, antiproliferative, antiarthritic, anticaries, and so on. OBJECTIVE: The objective of this current study is to give an overview of the recent literature and patents of T. chebula and chebulinic acid including methods of its isolation/extraction and their application in the prevention of various cancers and other diseases. METHODS: Present research and patents highlighting the anti-cancer potential of T. chebula and chebulinic acid have been studied and discussed keeping in view the scientific novelty and impact. RESULTS: Both T. chebula and chebulinic acid are currently being explored for their anticancer potential in vitro and in vivo. They are either incorporated alone or in combination with other plants or drugs to show their activity and many clinical trials are also going on various potentials of the plant and chebulinic acid. Novel extraction techniques are also explored and patented. Efforts are being made to improve the bioavailability by developing Novel herbal drug delivery systems of the plant extract or chebulinic acid itself. CONCLUSION: Anti-cancer potential of T. chebula and chebulinic acid may be well established by promising clinical trials and may open new interventions in various tumors. Clinical trials in conjunction with standard therapies are required to explore and validate the actual potential of T. chebula and chebulinic acid respectively.

Effect of combustion particle morphology on biological responses in a Co-culture of human lung and macrophage cells.

Atmospheric aging of combustion particles alters their chemical composition and morphology. Previous studies have reported differences in toxicological responses after exposure to fresh versus aged particles, with chemical composition being the prime suspect behind the differences. However, less is known about the contribution of morphological differences in atmospherically aged particles to toxicological responses, possibly due to the difficulty in resolving the two properties (composition and morphology) that change simultaneously. This study altered the shape of lab-generated combustion particles, without affecting the chemical composition, from fractal-like to a more compact spherical shape, using a water condensation-evaporation method. The two shapes were exposed to a co-culture of human airway epithelial (A549) and differentiated human monocyte (THP-1) cells at air-liquid interface (ALI) conditions. The particles with different shapes were deposited using an electrostatic field-based ALI chamber. For the same mass dose, both shapes were internalized by cells, induced a pro-inflammatory response (IL-8 and TNFα), and enhanced CYP1A1 gene expression compared to air controls. The more compact spherical particles (representative of atmospherically aged particles) induced more early apoptosis and release of TNFα compared to the more fractal-like particles. These results suggest a contribution of morphology to the increased toxicity of aged combustion-derived particles.

Comparison of biological responses between submerged, pseudo-air-liquid interface, and air-liquid interface exposure of A549 and differentiated THP-1 co-cultures to combustion-derived particles.

Air liquid interface (ALI) exposure systems are gaining interest, and studies suggest enhanced response of lung cells exposed to particles at ALI as compared to submerged exposure, although the results have been somewhat inconsistent. Previous studies have used monocultures and measured particle deposition using assumptions including consistent particle deposition, particle density, and shape. This study exposed co-cultures of A549 and differentiated THP-1 cells to flame-generated particles using three exposure methods: ALI, pseudo-ALI, and submerged. The dose at ALI was measured directly, reducing the need for assumptions about particle properties and deposition. For all exposure methods an enhanced pro-inflammatory response (TNFα) and Cytochrome P450 (CYP1A1) gene expression, compared to their corresponding negative controls, was observed. ALI exposure induced a significantly greater TNFα response compared to submerged exposure. The submerged exposures exhibited greater induction of CYP1A1 than other exposure methods, although not statistically significant. Some of the factors behind the observed difference in responses for the three exposure methods include differences in physicochemical properties of particles in suspending media, delivered dose, and potential contribution of gas-phase species to cellular response in ALI exposure. However, given the difficulty and expense of ALI exposures, submerged exposure may still provide relevant information for particulate exposures.

Nasal endoscopy, room filtration, and aerosol concentrations during live outpatient encounters: a prospective, case-control study.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic has highlighted safety concerns surrounding possible aerosol-generating procedures, but comparative data on the smallest particles capable of transmitting this virus remain limited. We evaluated the effect of nasal endoscopy on aerosol concentration and the role of a high-efficiency particulate air (HEPA) filter in reducing aerosol concentration. METHODS: Otolaryngology patients were prospectively enrolled in an outpatient, cross-sectional study. Demographic information and clinic room characteristics were recorded. A scanning mobility particle sizer and GRIMM aerosol monitor measured aerosols 14.3 nm to 34 µm in diameter (i.e., particles smaller than those currently examined in the literature) during (1) nasal endoscopy (± debridement) and (2) no nasal endoscopy encounters. One-way analysis of variance (ANOVA) and Student's t test were performed to compare aerosol concentrations and impact of HEPA filtration. RESULTS: Sixty-two patients met inclusion criteria (25 nasal endoscopy without debridement; 18 nasal endoscopy with debridement; 19 no nasal endoscopy). There was no significant difference in age or gender across cohorts. Aerosol concentration in the nasal endoscopy cohort (± debridement) was not greater than the no nasal endoscopy cohort (p = 0.36; confidence interval [95% CI], -1.76 to 0.17 µg/m3 ; and p = 0.12; 95% CI, -0.11 to 2.14 µg/m3 , respectively). Aerosol concentrations returned to baseline after 8.76 min without a HEPA filter versus 4.75 min with a HEPA filter (p = 0.001; 95% CI, 1.73-6.3 min). CONCLUSION: Using advanced instrumentation and a comparative study design, aerosol concentration was shown to be no greater during nasal endoscopy versus no endoscopy encounters. HEPA filter utilization reduced aerosol concentrations significantly faster than no HEPA filter.

Feasibility of a High-Volume Filter Sampler for Detecting SARS-CoV-2 RNA in COVID-19 Patient Rooms.

Aerosolization of SARS-CoV-2 by COVID-19 patients can put healthcare workers and susceptible individuals at risk of infection. Air sampling for SARS-CoV-2 has been conducted in healthcare settings, but methods vary widely and there is need for improvement. The objective of this study was to evaluate the feasibility of using a high-volume filter sampler, BioCapture z720, to detect SARS-CoV-2 in COVID-19 patient rooms in a medical intensive care unit, a dedicated COVID-19 ward, and at nurses' stations. In some locations, the BioSpot-VIVAS, known for high efficiency in the collection of virus-containing bioaerosols, was also operated. The samples were processed for SARS-CoV-2 RNA with multi-plex nested polymerase chain reaction. One of 28 samples collected with the high-volume filter sampler was positive for SARS-CoV-2; all 6 samples collected with BioSpot-VIVAS were negative for SARS-CoV-2. The high-volume filter sampler was more portable and less intrusive in patient rooms than the BioSpot-VIVAS, but limits of detection remain unknown for this device. This study will inform future work to evaluate the reliability of these types of instruments and inform best practices for their use in healthcare environments for SARS-CoV-2 air sampling.

GM-CSF production by non-classical monocytes controls antagonistic LPS-driven functions in allergic inflammation.

Lipopolysaccharide (LPS) can either promote or prevent T helper 2 (Th2) cell allergic responses. However, the underlying mechanism remains unknown. We show here that LPS activity switches from pro-pathogenic to protective depending on the production of granulocyte-macrophage colony-stimulating factor (GM-CSF) by non-classical monocytes. In the absence of GM-CSF, LPS can favor pathogenic Th2 cell responses by supporting the trafficking of lung-migratory dendritic cells (mDC2s) into the lung-draining lymph node. However, when non-classical monocytes produce GM-CSF, LPS and GM-CSF synergize to differentiate monocyte-derived DCs from classical Ly6Chi monocytes that instruct mDC2s for Th2 cell suppression. Importantly, only allergens with cysteine protease activity trigger GM-CSF production by non-classical monocytes. Hence, the therapeutic effect of LPS is restricted to allergens with this enzymatic activity. Treatment with GM-CSF, however, restores the protective effects of LPS. Thus, GM-CSF produced by non-classical monocytes acts as a rheostat that fine-tunes the pathogenic and therapeutic functions of LPS.

Determining real-time mass deposition with a quartz crystal microbalance in an electrostatic, parallel-flow, air-liquid interface exposure system.

In vitro studies are the first step toward understanding the biological effects of particulate matter. As a more realistic exposure strategy than submerged culture approaches, air-liquid interface (ALI) in vitro exposure systems are gaining interest. One challenge with ALI systems is determining accurate particle mass deposition. Although a few commercially available ALI systems are equipped with online mass deposition monitoring, most studies use indirect methods to estimate mass doses. These different indirect methods may contribute to inconsistencies in the results from in vitro studies of aerosolized nanoparticles. This study explored the effectiveness of using a commercially available Quartz Crystal Microbalance (QCM) to estimate the real-time, particle-mass deposition inside an electrostatic, parallel-flow, ALI system. The QCM system required minor modifications, including custom-designed and fabricated headers. Three QCM systems were simultaneously placed in three of the six wells in the ALI exposure chamber to evaluate the uniformity of particle deposition. The measurements from fluorescein dosimetry and QCM revealed an uneven deposition between these six wells. The performance of the QCM system was also evaluated using two different methods. First, using fluorescein deposition in one well, depositions in three other wells were estimated, which was then compared to the actual QCM readings. Second, using the QCM measured deposition in one well, the deposition in three other wells was estimated and compared to deposition measured by fluorescein dosimetry. For both methods, the expected and actual deposition yields a linear fit with the slope ~1. This good fit suggests that QCM systems can be used to measure real-time mass deposition in an electrostatic ALI system.

Dihydrotestosterone in Amyotrophic lateral sclerosis-The missing link?

OBJECTIVE: Testosterone has been postulated to be involved in ALS causation. MATERIALS AND METHODS: CSF levels of free testosterone and dihydrotestosterone were measured in 13 ALS patients [7 males, 6 females] and 22 controls [12 males, 10 females]. RESULTS: CSF free testosterone levels did not show any significant differences but CSF dihydrotestosterone levels were significantly decreased in all male and female ALS patients. CONCLUSIONS: DHT is probably integral to survival of motor neurons. In patients predisposed to develop ALS, there is possibly a sort of "testosterone resistance" at level of blood-brain barrier [BBB] existing right from birth and is likely the result of dysfunctional transport protein involved in testosterone transfer across the BBB. In these patients, lesser amount of testosterone is able to breach the BBB and enter the central neural axis. Lesser amount of testosterone is available to 5 α reductase in the anterior pituitary to be converted to DHT and lesser amount of DHT is generated. There is inadequate negative feedback suppression of LH at the level of anterior pituitary by DHT. As a result of higher LH levels, testosterone levels rise in the peripheral testosterone fraction [the fraction outside the BBB] and this explains the various physical attributes of ALS patients like lower Ratio of the index and ring finger lengths (2D:4D ratio), increased incidence of early onset alopecia etc. This deficiency of DHT leads to motor neuron death causing ALS.

Characterization of renewable diesel particulate matter gathered from non-premixed and partially premixed flame burners and from a diesel engine.

Particulate matter coming from the combustion of renewable diesel (RD), ultra-low sulfur diesel (ULSD) and a volumetric blend of 30% of RD with ULSD (RD30) were collected and physico-chemically characterized. Soot samples were generated in two flame burner types (non-premixed flame, NPF, and partially premixed flame, PPF) trying to simulate the diffusion and premix regimes found in diesel engines. The impact of both fuel nature and burner type was assessed on soot mass, particle size and morphology, particle nanostructure and surface functional groups. In general, although the results of HRTEM and SMPS suggested that the addition of RD reduced the average particle size and increased the concentration of ultra-fine particles, the mass emission of soot was drastically mitigated regardless of the burner used. The results also suggest that the changes in the chemical characteristics of the soot were slightly more sensitive than the changes in the internal nanostructure of the particles, since the graphitic character (as showed by Raman and infrared analysis) increased as the RD content increased, being stronger for the PPF system. Comparisons between engine soot and flame soot confirmed that the addition of RD into ULSD produced smaller and more carbonized particles. In fact, some engine results were located in between those obtained in PPF and NPF burners, suggesting that both combustion regimes are contributing to soot characteristics in engines. This consistency suggests that a first assessment of the impact of alternative fuels on the characteristics of particulate matter can be conducted through the basic approach offered by laboratory flames, thereby avoiding the costs associated with generating large quantities of fuel and the complexities of in-cylinder physical interactions and engine parameters.

Effect of collection methods on combustion particle physicochemical properties and their biological response in a human macrophage-like cell line.

In vitro studies are a first step toward understanding the biological effects of combustion-derived particulate matter (cdPM). A vast majority of studies expose cells to cdPM suspensions, which requires a method to collect cdPM and suspend it in an aqueous media. The consequences of different particle collection methods on particle physiochemical properties and resulting biological responses are not fully understood. This study investigated the effect of two common approaches (collection on a filter and a cold plate) and one relatively new (direct bubbling in DI water) approach to particle collection. The three approaches yielded cdPM with differences in particle size distribution, surface area, composition, and oxidative potential. The directly bubbled sample retained the smallest sized particles and the bimodal distribution observed in the gas-phase. The bubbled sample contained ∼50% of its mass as dissolved species and lower molecular weight compounds, not found in the other two samples. These differences in the cdPM properties affected the biological responses in THP-1 cells. The bubbled sample showed greater oxidative potential and cellular reactive oxygen species. The scraped sample induced the greatest TNFα secretion. These findings have implications for in vitro studies of air pollution and for efforts to better understand the underlying mechanisms.

Impaired Tumor-Necrosis-Factor-α-driven Dendritic Cell Activation Limits Lipopolysaccharide-Induced Protection from Allergic Inflammation in Infants.

Infants have a higher risk of developing allergic asthma than adults. However, the underlying mechanism remains unknown. We show here that sensitization of mice with house-dust mites (HDMs) in the presence of low-dose lipopolysaccharide (LPS) prevented T helper 2 (Th2) cell allergic responses in adult, but not infant, mice. Mechanistically, adult CD11b+ migratory dendritic cells (mDCs) upregulated the transcription factor T-bet in response to tumor necrosis factor-α (TNF-α), which was rapidly induced after HDM + LPS sensitization. Consequently, adult CD11b+ mDCs produced interleukin-12 (IL-12), which prevented Th2 cell development by promoting T-bet upregulation in responding T cells. Conversely, infants failed to induce TNF-α after HDM + LPS sensitization. Therefore, CD11b+ mDCs failed to upregulate T-bet and did not secrete IL-12 and Th2 cell responses normally developed in infant mice. Thus, the availability of TNF-α dictates the ability of CD11b+ mDCs to suppress allergic Th2-cell responses upon dose-dependent endotoxin sensitization and is a key mediator governing susceptibility to allergic airway inflammation in infant mice.

Application of a Quartz Crystal Microbalance to Measure the Mass Concentration of Combustion Particle Suspensions.

Researchers studying the biological effects of combustion particles typically rely on suspending particles in de-ionized (DI) water, buffer, and/or media prior to in vitro or in vivo experiments. However, the hydrophobic nature of combustion particles makes it difficult to obtain well-suspended, evenly dispersed mixtures, which also makes it difficult to obtain equivalent dosing and endpoint comparisons. This study explored the use of a quartz crystal microbalance (QCM) to measure the mass concentration of combustion particle suspensions. It compared the QCM mass concentration to that estimated by placing a known mass of combustion particles in DI water. It also evaluated the effect of drop volume and combustion particle type on QCM measurements. The results showed that QCM is a promising direct method for measuring suspended combustion particle mass concentrations, and it is particularly effective for quantifying concentrations of difficult-to-suspend particles and for combustion particles placed in polystyrene containers, which can lead to substantial particle losses.

Antibiotic-mediated bacteriome depletion in ApcMin/+ mice is associated with reduction in mucus-producing goblet cells and increased colorectal cancer progression.

Recent epidemiological evidence suggests that exposure to antibiotics in early-to-middle adulthood is associated with an increased risk of colorectal adenoma. However, mechanistic studies in established preclinical cancer to examine these claims are extremely limited. Therefore, we investigated the effect of long-term exposure of an antibiotic cocktail composed of Vancomycin, Neomycin, and Streptomycin, on tumor development and progression in the ApcMin/+ mouse, an established genetic model for familial adenomatous polyposis. Clinical pathologies related to tumor development as well as intestinal and colon tissue histopathology were studied at ages 8, 12, and 16 weeks of age, which correspond to the approximate ages of development of neoplasia, gut inflammation with polyposis, and cancer progression, respectively, in this animal model. We show that the antibiotics significantly increase the severity of clinical symptoms, including effects on intestinal histology and goblet cell numbers. In addition, they promote small intestinal polyposis. Finally, metagenomic analysis of fecal samples demonstrated that antibiotic exposure is associated with a significant but nonuniform depletion of the animal's natural gut flora. Overall, these findings support the premise that long-term antibiotic exposure mediates the selected depletion of gut microbial communities and the concomitant thinning of the protective mucus layer, resulting in an increase in tumor development.

Chemotherapy with si-RNA and Anti-Cancer Drugs.

BACKGROUND: To treat cancer, chemotherapy is a key therapeutic approach which is associated with several limitations. This chemotherapeutical agent exhibits multi drug resistance coupled with undesirable side effects. This multidrug resistance is exhibited by tumor cell due to actuation of drug out flow mechanism, programmed cell death and protection mechanisms etc. One of the therapeutic approaches to cure cancer is RNA interference (RNAi). Small interfering RNA (si-RNA) is considered as a major therapeutic tool used to control expression of a particular gene. It is a well known fact that intake of more drugs can lead to cancer chemo resistance, thus siRNA based therapeutic approach is under scrutiny to cure cancer. METHODS: This review article gives an overview of various combination approaches for si-RNA with chemotherapeutics. Further, article highlights the potential of nanotechnology to improve bioavailability of drug and bio-therapeutics at the site of action. RESULTS: Combination chemotherapy is employed in clinics as a main cancer treatment tool to suppress multidrug resistance in cancer. On the other hand, suitable protective carrier is needed due to the stability issues and small size of si-RNA. To overcome these drawbacks associated with siRNA currently, nanotechnology based approaches have been widely used. CONCLUSION: Delivery of anti-cancer drugs with si-RNA will be one of important intermingled approach to reduce duration of chemotherapy and improve therapeutic outcomes in cancer patient.

Dietary selenium protects adiponectin knockout mice against chronic inflammation induced colon cancer.

Selenium (Se) is an essential dietary micronutrient that has been examined for protection against different types of cancers including colon cancer. Despite an established inverse association between Se and chronic inflammation induced colon cancer (CICC), the mechanistic understanding of Se's protective effects requires additional in-vivo studies using preclinical animal models of CICC. Adiponectin (APN) is an adipocytokine that is protective against CICC as well. However, its role in the anti-mutagenic effects of the Se-diet remains unknown. To address this knowledge gap, here we examine the ability of dietary Se in reducing CICC in APN knockout mice (KO) and its wild-type C57BL/6. CICC was induced with the colon cancer agent 1,2 dimethyl hydrazine (DMH) along with dextran sodium sulfate (DSS). Se-enhanced diet increased selenoproteins, Gpx-1 and Gpx-2, in the colon tissues, thereby reducing oxidative stress. Se-mediated reduction of CICC was evident from the histopathological studies in both mouse models. In both mice, reduction in inflammation and tumorigenesis associated well with reduced p65 phosphorylation and elevated 53 phosphorylation. Finally, we show that in both models Se-administration promotes goblet cell differentiation with a concomitant increase in the levels of associated proteins, Muc-2 and Math-1. Our findings suggest that Se's protection against CICC involves both colonic epithelial protection and anti-tumor effects that are independent of APN.

Fluoroquinolone-related neuropsychiatric and mitochondrial toxicity: a collaborative investigation by scientists and members of a social network.

BACKGROUND: The 3 fluoroquinolone (FQ) antibiotics - ciprofoxacin, levofoxacin, and moxifoxacin - are commonly administered to oncology patients. Although these oral antibiotics are approved by the US Food and Drug Administration (FDA) for treatment of urinary tract infections, acute bacterial sinusitis, or bacterial infection in patients with chronic obstructive pulmonary disease, they are commonly prescribed off-label to neutropenic cancer patients for the prevention and treatment of infections associated with febrile neutropenia. New serious FQ-associated safety concerns have been identified through novel collaborations between FQ-treated persons who have developed long-term neuropsychiatric (NP) toxicity, pharmacovigilance experts, and basic scientists. OBJECTIVE: To conduct basic science and clinical investigations of a newly identified adverse drug reaction, termed FQ-associated disability. METHODS: 5 groups of C57BL/6 mice receiving the antibiotic ciprofoxacin in 10-mg increments (10 mg/kg-50 mg/kg) and 1 group of control mice were evaluated. The Southern Network on Adverse Reactions (SONAR) and a social network of FQ-treated persons with long-term NP toxicity (the Floxed Network) conducted a web-based survey. The clinical toxicity manifestations reported by 94 respondents to the web-based survey of persons who had received 1 or more doses of an FQ prescribed for any indication (generally at FDA-approved dosages) and who subsequently experienced possible adverse drug reactions were compared with adverse event information included on the product label for levofoxacin and with FQ-associated adverse events reported to the FDA's MedWatch program. RESULTS: Mice treated with ciprofoxacin had lower grip strengths, reduced balance, and depressive behavior compared with the controls. For the survey, 93 of 94 respondents reported FQ-associated events including anxiety, depression, insomnia, panic attacks, clouded thinking, depersonalization, suicidal thoughts, psychosis, nightmares, and impaired memory beginning within days of FQ initiation or days to months of FQ discontinuation. The FDA Adverse Event Reporting System (FAERS) included 210,705 adverse events and 2,991 fatalities for FQs. Levofoxacin and ciprofoxacin toxicities were neurologic (30% and 26%, respectively), tendon damage (8% and 6%), and psychiatric (10% and 2%). In 2013, an FDA safety review reported that FQs affect mammalian topoisomerase II, especially in mitochondria. In 2013 and 2014, SONAR fled citizen petitions requesting black box revisions identifying neuropsychiatric toxicities and mitochrondrial toxicity as serious levofoxacin-associated adverse drug reactions. In 2015, FDA advisors recommended that FQ product labels be revised to include information about this newly identified disability syndrome termed "FQ-associated disability" (FQAD). LIMITATIONS: Basic science studies evaluated NP toxicity for only 1 FQ, ciprofoxacin. CONCLUSION: Pharmacovigilance investigators, a social network, and basic scientists can collaborate on pharmacovigilance investigations. Revised product labels describing a new serious adverse drug reaction, levofoxacin-associated long-term disability, as recommended by an FDA advisory committee, are advised.

Short-Term Moderate Sleep Restriction Decreases Insulin Sensitivity in Young Healthy Adults.

CONTEXT AND PURPOSE: The literature suggests that severe sleep loss of more than a few hours a night decreases glucose tolerance and insulin sensitivity. The aim of this study was to determine whether moderate sleep restriction had similar effects. METHODS: Fifteen healthy non-obese (BMI=24.5±3.4 kg/m2) young adults (20.6±1.3 years) completed two 2-hour oral glucose tolerance tests (OGTT): one was after 3 days of time-in-bed restriction by 1-3 hours each night, and the other was after 3 days of ad libitum sleep. Glucose and insulin concentrations during OGTT, and fasting glucagon and cortisol concentrations were determined. The homeostasis model of insulin resistance (HOMA-IR), Matsuda index, and the quantitative insulin sensitivity check index (QUICKI) were calculated. RESULTS: The total time-in-bed during the sleep restriction and the ad libitum phase was 5.98±0.76 and 7.98±0.54 hours/day, and total sleep time was 5.16±0.49 and 6.65±0.64 hours/day, respectively. Glucose concentrations before and 30, 60, 90, and 120 minutes following consumption of glucose and area under the curve were not different for the two OGTT (p > 0.10 for all). Insulin concentration at fasting and area under the curve during the OGTT were significantly higher (p = 0.034 and 0.038, respectively) following restricted sleep than following ad libitum sleep. Fasting glucagon concentration was also higher (p = 0.003). The HOMA-IR, Matsuda index, and QUICKI all suggested decreased insulin sensitivity following restricted sleep. CONCLUSION: Short-term moderate sleep restriction reduced insulin sensitivity compared to ad libitum sleep in this group of healthy young adults.

Mucus mediated protection against acute colitis in adiponectin deficient mice.

BACKGROUND: Acute ulcerative colitis is an inflammation-driven condition of the bowel. It hampers the general homeostasis of gut, resulting in decreased mucus production and epithelial cell renewal. Adiponectin (APN), an adipocytokine, is secreted by the adipose tissue and has been debated both as a pro-inflammatory or anti-inflammatory protein depending on the disease condition and microenvironment. The present study delineates the role of APN depletion in mucus modulation in a model of acute colitis. METHODS: APNKO and C57BL/6 (WT) male mice were given 2% DSS ad libidum for 5 days in drinking water, followed by normal drinking water for the next 5 days. Hematoxyline-eosin and Alcian Blue staining was used to observe the general colonic morphology and goblet cell quantification respectively. Protein expression levels were quantified by Western blot for MATH1, Hes1, MUC2 and MUC4. ELISA was used to study the levels of TNF-α, IL-6 and IL-1ß. RESULTS: APNKO mice showed significantly higher goblet to epithelial cell ratios, lower pro-inflammatory cytokines and higher MUC2 levels as compared to the WT mice. The protein expression levels for the mucin MUC2 supported the histopathological findings. An increase in colon tissue-secreted levels of pro-inflammatory with a reduction in anti-inflammatory cytokines in presence of APN support the pro-inflammatory role of APN during acute inflammation. CONCLUSION: Absence of APN is protective against DSS-induced acute colonic inflammation by means of reducing colon tissue-secreted pro-inflammatory cytokines, modulating goblet and epithelial cell expressions, and increasing the levels of secretory mucin MUC2.

Stealth lipid coated aquasomes bearing recombinant human interferon-α-2b offered prolonged release and enhanced cytotoxicity in ovarian cancer cells.

PURPOSE: In present investigation, recombinant human interferon-α-2b (rhINF-α-2b) loaded aquasomes were prepared, optimized and overlaid with PEGylated phospholipid to offer prolong release and high therapeutic index against ovarian cancer, SKOV3 cells. METHODS AND RESULTS: Central Composite Design (CCD) and Response Surface Methodology (RSM) were employed to calculate the optimized conditions, 1:3 core to coat ratio, sonication power of 12.5W and time of about 55min for preparation of aquasomes. Consequently, rhINF-α-2b-Py-5-P-Aq.somes exhibited higher protein loading capacity and retained structural conformations of rhINF-α-2b, as compared to rhINF-α-2b-Cellob-Aq.somes, rhINF-α-2b-Tre-Aq.somes and rhINF-α-2b-Core (CaHPO4). Further, optimized rhINF-α-2b-Py-5-P-Aq.somes was superimposed with phospholipid-PEG2000 to prolong the release pattern of rhINF-α-2b from aquasomes. The rhINF-α-2b-core (CaHPO4) released 97.3% of protein in 1h, while 95.3% of rhINF-α-2b was released by rhINF-α-2b-Tre-Aq.somes in 4h. Concurrently, rhINF-α-2b-Cellob-Aq.somes and rhINF-α-2b-Py-5-P-Aq.somes released 96.2% and 97.8% of rhINF-α-2b respectively in 6 and 8h. Ultimately, rhINF-α-2b-Py-5-P-Aq.somes-P-PEG2000 displayed evidence of its prolonged release pattern and released 98.1% of rhINF-α-2b in 336h. FT-IR and XRD substantiated the involvement of vigorous intermolecular hydrogen bonding and amorphous geometry in rhINF-α-2b-Py-5-P-Aq.somes. In last, rhINF-α-2b-Py-5-P-Aq.somes-P-PEG2000 exhibited the∼4.55, 1.92, 2.3, 2.8, and 3.84 fold reductions in IC50 as compared to free rhINF-α-2b, rhINF-α-2b-Py-5-P-Aq.somes, rhINF-α-2b-Cellob-Aq.somes, rhINF-α-2b-Tre-Aq.somes and rhINF-α-2b-Core (CaHPO4), respectively. CONCLUSION: Therefore, rhINF-α-2b-Py-5-P-Aq.somes-P-PEG2000 warrant further in depth in vitro and in vivo antitumor study to scale up the technology for clinical intervention.