Interval delivery of 5HT2A agonists using multilayered polymer films.

There is an urgent unmet medical need to develop therapeutic options for the ~50% of depression patients suffering from treatment-resistant depression, which is difficult to treat with existing psycho- and pharmaco-therapeutic options. Classical psychedelics, such as the 5HT2A agonists, have re-emerged as a treatment paradigm for depression. Recent clinical trials highlight the potential effectiveness of 5HT2A agonists to improve mood and psychotherapeutic growth in treatment-resistant depression patients, even in those who have failed a median of four previous medications in their lifetime. Moreover, microdosing could be a promising way to achieve long-term alleviation of depression symptoms without a hallucinogenic experience. However, there are a gamut of practical barriers that stymie further investigation of microdosing 5HT2A agonists, including: low compliance with the complicated dosing regimen, high risk of diversion of controlled substances, and difficulty and cost administering the long-term treatment regimens in controlled settings. Here, we developed a drug delivery system composed of multilayered cellulose acetate phthalate (CAP)/Pluronic F-127 (P) films for the encapsulation and interval delivery of 5HT2A agonists from a fully biodegradable and biocompatible implant. CAPP film composition, thickness, and layering strategies were optimized, and we demonstrated three distinct pulses from the multilayered CAPP films in vitro. Additionally, the pharmacokinetics and biodistribution of the 5HT2A agonist 2,5-Dimethoxy-4-iodoamphetamine (DOI) were quantified following the subcutaneous implantation of DOI-loaded single and multilayered CAPP films. Our results demonstrate, for the first time, the interval delivery of psychedelics from an implantable drug delivery system and open the door to future studies into the therapeutic potential of psychedelic delivery.

Chikungunya outbreak in Bangladesh (2017): sociodemographic and clinical characteristics of patients from three hotspots.

BACKGROUND: Chikungunya is a severely debilitating disease. Bangladesh witnessed one of the largest outbreaks in 2017. Here, we described the clinical profile of the chikungunya outbreak in Bangladesh and its heterogeneity across three hotspots. METHODS: This was a descriptive cross-sectional study of 432 individuals interviewed from the outpatient department of three study sites (Dhaka, Chittagong, and Sitakundu Upazilla of Bangladesh) after confirmation by the study physicians. Both laboratory-confirmed cases and probable cases were recruited between July and October 2017. RESULTS: Of all, 18% (79) were laboratory confirmed, and 353 82% (335) were probable cases. The male:female ratio was almost equal (1.09:1), and the predominant age group was 18-59 years. The mean age of the presentation was 36.07 ± 13.62 (SD) years. Fever and arthralgia were the most common presentations and were present in > 95% of cases. Other frequent symptoms were fatigue, myalgia, headache, nausea, and vomiting. Approximately half of the patients had arthritis and erythematous rash. Arthritis was predominant in Chittagong city, while maculopapular rash was not observed in Sitakunda city. However, fatigue, nausea, and vomiting are more common among patients in Dhaka city. Significant heterogeneity of clinical manifestations was present across the three hotspots (p < 0.05 for all). Both confirmed and probable cases shared similar characteristics except muscle ache (p = 0.22) and rash (p = 0.37). CONCLUSION: The clinical profile of chikungunya virus-induced disease displays significant location-related heterogeneity in Bangladesh during a large outbreak. Although the causes of such differences are unclear, improved public and medical personnel education on this condition may lead to earlier diagnosis and treatment.

ExoPRIME: Solid-phase immunoisolation and OMICS analysis of surface-marker-specific exosomal subpopulations.

Exosomes encapsulate genomic and proteomic biomarkers for non-invasive diagnosis and disease monitoring. However, exosome surface-markers heterogeneity is a major drawback of current isolation methods. Here, we report a direct, one-step exosome sampling technology, ExoPRIME, for selective capture of CD63+ exosome subpopulations using an immune-affinity protocol. Microneedles (300µm × 30 mm), functionalized with anti-CD63 antibodies, were incubated under various experimental conditions in conditioned astrocyte medium and astrocyte-derived exosome suspension. The probe's capture efficiency and specificity were validated using FluoroCet assay, immunofluorescent imaging, and OMICS analyses. Significantly higher exosomes were captured by probes incubated for 16 h at 4 0C in enriched exosomal suspension (23 × 10 6 exosomes per probe) vis-à-vis 2 h at 4 0 C (12 × 10 6) and 16 h at 22 0C (3 × 10 6) in conditioned cell media. Our results demonstrate the application of ExoPRIME over a broad dynamic range of temperature and incubation parameters, offering flexibility for any desired application. ExoPRIME permits the use and re-use of minimal sample volumes (≤200 µL), can be multiplexed in arrays, and integrated into a lab-on-a-chip platform to achieve parallel, high-throughput isolation of different exosome classes in a semi-automated workstation. This platform could provide direct exosomal analysis of biological fluids since it can elegantly interface with existing room-temperature, picomolar-range nucleic acid assays to provide a clinical diagnostic tool at the point of care.

Advances in Biosensors Technology for Detection and Characterization of Extracellular Vesicles.

Exosomes are extracellular vehicles (EVs) that encapsulate genomic and proteomic material from the cell of origin that can be used as biomarkers for non-invasive disease diagnostics in point of care settings. The efficient and accurate detection, quantification, and molecular profiling of exosomes are crucial for the accurate identification of disease biomarkers. Conventional isolation methods, while well-established, provide the co-purification of proteins and other types of EVs. Exosome purification, characterization, and OMICS analysis are performed separately, which increases the complexity, duration, and cost of the process. Due to these constraints, the point-of-care and personalized analysis of exosomes are limited in clinical settings. Lab-on-a-chip biosensing has enabled the integration of isolation and characterization processes in a single platform. The presented review discusses recent advancements in biosensing technology for the separation and detection of exosomes. Fluorescent, colorimetric, electrochemical, magnetic, and surface plasmon resonance technologies have been developed for the quantification of exosomes in biological fluids. Size-exclusion filtration, immunoaffinity, electroactive, and acoustic-fluid-based technologies were successfully applied for the on-chip isolation of exosomes. The advancement of biosensing technology for the detection of exosomes provides better sensitivity and a reduced signal-to-noise ratio. The key challenge for the integration of clinical settings remains the lack of capabilities for on-chip genomic and proteomic analysis.

Analysis of the dark proteome of Chandipura virus reveals maximum propensity for intrinsic disorder in phosphoprotein.

Chandipura virus (CHPV, a member of the Rhabdoviridae family) is an emerging pathogen that causes rapidly progressing influenza-like illness and acute encephalitis often leading to coma and death of the human host. Given several CHPV outbreaks in Indian sub-continent, recurring sporadic cases, neurological manifestation, and high mortality rate of this infection, CHPV is gaining global attention. The 'dark proteome' includes the whole proteome with special emphasis on intrinsically disordered proteins (IDP) and IDP regions (IDPR), which are proteins or protein regions that lack unique (or ordered) three-dimensional structures within the cellular milieu. These proteins/regions, however, play a number of vital roles in various biological processes, such as cell cycle regulation, control of signaling pathways, etc. and, therefore, are implicated in many human diseases. IDPs and IPPRs are also abundantly found in many viral proteins enabling their multifunctional roles in the viral life cycles and their capability to highjack various host systems. The unknown abundance of IDP and IDPR in CHPV, therefore, prompted us to analyze the dark proteome of this virus. Our analysis revealed a varying degree of disorder in all five CHPV proteins, with the maximum level of intrinsic disorder propensity being found in Phosphoprotein (P). We have also shown the flexibility of P protein using extensive molecular dynamics simulations up to 500 ns (ns). Furthermore, our analysis also showed the abundant presence of the disorder-based binding regions (also known as molecular recognition features, MoRFs) in CHPV proteins. The identification of IDPs/IDPRs in CHPV proteins suggests that their disordered regions may function as potential interacting domains and may also serve as novel targets for disorder-based drug designs.

A phase I delayed-start, randomized and pharmacodynamic study of metformin and chemotherapy in patients with solid tumors.

PURPOSE: Metformin activates AMP-related pathways leading to inactivation of mammalian target of rapamycin (mTOR) and suppression of its downstream effectors, crucial for cancer growth. Epidemiologic studies showed a reduced incidence and improved survival in cancer patients. We conducted a prospective phase I study to assess the safety of metformin in combination with chemotherapy in patients with solid tumors. METHODS: We conducted a delayed-start randomized trial of non-diabetic patients in two stages. In Stage 1, we randomized patients to two arms: concurrent arm (metformin with chemo) vs. delayed arm (chemo alone). In Stage 2, patients in delayed arm were crossed over to receive metformin. Patients received metformin 500 mg twice daily with chemotherapy to define dose-limiting toxicities (DLTs) in both stages. Secondary endpoints assessed adverse events (AEs) and response rates. Translational correlates included effects of metformin on expression and phosphorylation of 5' adenosine monophosphate-activated protein kinase (AMPK) by western blot in PBMCs. RESULTS: A total of 100 patients were enrolled (51 in delayed arm vs. 49 concurrent arm). Rate of DLTs in patients receiving metformin with chemotherapy was 6.1% vs. 7.8% in patients receiving chemotherapy alone. DLTs seen with addition of metformin included those associated with established chemo adverse events. No lactic acidosis or hypoglycemia occurred. Restaging showed stable disease in 46% at cessation of metformin. 28% of patients with measurable tumor markers showed improvement. AMPK phosphorylation showed a four- to sixfold increase in AMPK phosphorylation after metformin. CONCLUSIONS: This is the largest phase I study of metformin combined with chemotherapy, which suggests that metformin can be given safely with chemotherapy, and offers a platform for future studies. Post-metformin increase in AMPK phosphorylation may potentially explain lack of disease progression in nearly half of our patients. FUNDING: UL1 TR001064. CLINICAL TRIAL INFORMATION: NCT01442870.

Calorimetric sandwich-type immunosensor for quantification of TNF-α.

We report a lab-on-a-chip immunosesnor for quantification of the inflammatory cytokine TNF-α with picomolar sensitivity. The feasibility of the technology was demonstrated via accurate measurement of the concentration of TNF-α in astrocytes cell culture media. The immunoassay was performed in a microfluidic device with an integrated antimony/bismuth thermopile sensor and had a limit of detection of 14 pg mL-1. The device was fabricated using rapid prototyping xurography technique and consisted of two inlets and single outlet. Anti-TNF-α monoclonal antibody was used to capture the analyte while the detection was performed using glucose oxidase-conjugated secondary antibody. Glucose (55 mM) was injected through a sample loop into the fluid flowing within the microfluidic device. A nanovolt meter connected to the thermoelectric sensor recorded the voltage change caused by the enzymatic reaction. Computer simulations using COMSOL Multiphysics were performed to analyze the effect of fluid velocity on the concentration of glucose within the reaction zone. A standard calibration curve was created using serial dilutions of synthetic TNF-α (0-2000 pg mL-1) by plotting the area under the curve of the signal versus the concentration of the analyte. The efficacy of the device was evaluated by quantifying TNF-α in the cell culture medium of lipopolysaccharide stimulated and non-stimulated astrocytes. The results demonstrated high accuracy of the calorimetric immunoassay when compared with gold standard commercial ELISA microplate reader. The immunosensor offers excellent reproducibility, accuracy, and versatility in the choice of the detection enzyme.

Binding of cadmium to Strychnos potatorum seed proteins in aqueous solution: adsorption kinetics and relevance to water purification.

Strychnos potatorum seeds (cleaning nuts or nirmali) are extensively used by remote village tribals in the state of Andhra Pradesh, India for clarification of turbid and metal contaminated water. In the present study the ability of seed proteins to bind aqueous cadmium has been investigated. Biochemical characterization of the seed powder revealed the presence of coagulant proteins. These proteins were isolated from the soluble extracts of the seeds by ammonium sulfate fractionation. The (30-70%) fraction containing the bulk of proteins were separated by gel filtration into two peaks A and B. The (30-70%) ammonium sulfate precipitated proteins, as well as those from Peak A and B were separately immobilized to affigel-10. The Cd(II) biosorption efficiency by these proteins have been investigated. Different experiments have been conducted (i) over a range of pH (2.0-7.0), (ii) contact time (5-600 min), (iii) temperatures (4-40°C) and (iv) metal ion concentrations (80-110 mg L(-1)). The results showed that the optimum conditions for Cd(II) adsorption are almost same for the three proteins used in the study. Cd(II) removal is pH dependent and the maximum removal was at pH 5.0 in a time span of 360 min. The equilibrium data fit into Langmuir isotherm than Freundlich model. The correlation coefficient for the pseudo second order is high (~0.996-1.00) where as the correlation coefficient of the pseudo first order model is too low so the adsorption is better described by pseudo second order model.

Subjective sleep duration and quality influence diet composition and circulating adipocytokines and ghrelin levels in teen-age girls.

Understanding the interplay between sleep duration and quality, diet and hormones of obesity may help design effective lifestyle intervention strategies. Here we studied such associations in lean and obese teen-aged Saudi girls. In this cross-sectional observational study, 126 girls (62 lean and 64 obese) aged 14 -18 years (16.5 ± 1.5) were evaluated. A general questionnaire, which included sleep and diet questions, was obtained and anthropometric measurements and overnight fasting blood samples for determination of glucose, lipid profile and serum levels of leptin, adiponectin, resistin and ghrelin were collected. Subjects that slept < 5 hours/day had a higher percent of carbohydrate intake (p = 0.04) than those who slept > 7 hours/day. Adiponectin levels were higher in the lean than the obese group and increased in proportion to hours of sleep. Ghrelin had an inverse association with subjective sleep duration (p = 0.04), while resistin levels were directly proportional to it. Thus, the duration and quality of sleep influenced diet composition and the circulating levels of adipocytokines and ghrelin in adolescent girls. Long and uninterrupted sleep was associated with a better diet and a more favorable hormonal profile.

Role of panitumumab in the management of metastatic colorectal cancer.

Panitumumab (formerly known as ABX-EGF) is the first fully human monoclonal antibody directed against the epidermal growth factor receptor in clinical use. It has proven to be very well tolerated alone and in combination with other cytotoxic chemotherapeutic agents. Panitumumab has demonstrated efficacy as monotherapy and with standard chemotherapeutic agents in a wide variety of cancer types, including non-small-cell lung cancer, renal, and colorectal cancer (CRC). To date, no human antihuman antibodies have been detected, and unlike cetuximab, infusion reactions are infrequent, and no premedications are required when administering panitumumab. The only significant toxicity has been a rash similar to that seen with other agents targeting the epidermal growth factor receptor, and such reactions have been predominantly mild to moderate. In metastatic CRC, panitumumab has been safe and efficacious when given with other commonly used agents in this disease, including irinotecan and fluorouracil. Current studies under way are looking at panitumumab in combination with FOLFOX (fluorouracil/leucovorin/oxaliplatin) plus bevacizumab as well as with novel agents that have yet to come into common clinical practice. Recent progress in development of panitumumab in the management of CRC is reviewed, and management of associated rash is discussed herein.

Targeted agents for adjuvant therapy of colon cancer.

Among patients with colorectal cancer (CRC) diagnosed in the United States, 37.2% are diagnosed with stage III and 27.9% with stage II disease. In locoregionally advanced CRC, surgery is the primary treatment modality and has a curative intent. The survival depends on the pathologic stage and varies from 30%-60% for stage III to 60%-80% for stage II. However, as much as 40%-50% of patients will relapse and require additional treatment of the disease. Clinical failure after resection of CRC is predominantly secondary to the clinical progression of previously undetected distant metastatic disease. Until very recently, the absolute benefit for survival obtained with adjuvant therapy compared with control was about 6%. Introduction of oxaliplatin in the adjuvant setting has shown a reduction of 23% in the risk of relapse when compared with 5-fluorouracil alone (MOSAIC). Recent phase III studies have shown that targeted agents improved survival in patients with advanced-stage CRC. Bevacizumab, a monoclonal antibody targeting vascular endothelial growth factor, is the first antiangiogenic drug to show improved efficacy when used in combination with irinotecan and oxaliplatin for first- and second-line treatment of CRC. Cetuximab, another monoclonal antibody targeting epidermal growth factor receptor, has shown efficacy in third-line therapy and promising results in first-line phase II studies. There is great interest in whether the biologic agents bevacizumab and cetuximab can improve survival in the adjuvant-therapy setting. This article reviews the adjuvant therapy for colon cancer and discusses the potential role and current trials involving the targeted agents.