Design and development of PEGylated liposomal formulation of HER2 blocker Lapatinib for enhanced anticancer activity and diminished cardiotoxicity.

Breast cancer is most frequently diagnosed cancer and fifth leading cause of death in women. About 20-30% of all breast cancers overexpress HER2/neu receptors. Lapatinib is a dual tyrosin kinase inhibitor of EGFR and HER2. It exhibits its anticancer effect via blocking intracellular domain of HER2 receptor in breast cancer. Lapatinib belongs to class II of BSC classification due to its poor solubility restricting its clinical application. Due to presence of HER2 receptor on cardiomyocytes, it is associated with generation of cardiotoxicity. The present study was aimed to design a PEGylated liposomal formulation of Lapatinib and evaluate its anticancer potential. Lapatinib liposomes were prepared using lipid layer hydration method and its characterization was done by determining its particle size, zeta potential, entrapment efficiency and in vitro release profiling. The anti-tumor activity of PEGylated liposomal formulation was evaluated in xenografted tumor induced by MDA-MB-453 breast cancer cells in chick embryos. The anti-tumor effect of lapatinib was enhanced by its PEGylated liposomal preparation as it led to the reduction in tumor size to a greater extent compared to the embryos treated with free lapatinib. Flowcytometric analysis and immunoflurescence study using cleaved PARP antibody demonstrated the enhanced apoptotic potential of PEGylated liposomes of lapatinib. SGOT levels, marker for cardiotoxicity and hepatotoxicity, significantly decreased in serum of embryos treated with PEGylated liposomes of lapatinib compared to free drug treated embryos. Hence, the PEGylated liposomal formulation of lapatinib can be used as a therapeutic strategy against HER2 positive breast cancer either alone or in combination with conventional anticancer agents and hormonal therapies.

Water-soluble prodrugs of paclitaxel containing self-immolative disulfide linkers.

A new series of disulfide-containing prodrugs of paclitaxel were designed, synthesized and evaluated against 6 cancer cell lines. Some of these prodrugs exhibited nearly equal or slightly better anticancer activity when compared to that of paclitaxel. These prodrugs contain water-soluble groups such as amino, carboxyl, hydroxyl, amino acids, etc., and exhibited 6-140 fold increase in aqueous solubility when compared to paclitaxel. One of these prodrugs exhibited improved water solubility, better in vitro anticancer activity and significantly superior oral bioavailability in mice when compared to those of paclitaxel. Thus, we have identified a very promising lead compound for further optimization and evaluation as a potentially bioavailable water-soluble prodrug of paclitaxel.

Impact of ALD grown passivation layers on silicon nitride based integrated optic devices for very-near-infrared wavelengths.

A CMOS compatible post-processing method to reduce optical losses in silicon nitride (Si(3)N(4)) integrated optical waveguides is demonstrated. Using thin layer atomic layer deposition (ALD) of aluminum oxide (Al(2)O(3)) we demonstrate that surface roughness can be reduced. A 40 nm thick Al(2)O(3) layer is deposited by ALD over Si(3)N(4) based strip waveguides and its influence on the surface roughness and the waveguide loss is studied. As a result, an improvement in the waveguide loss, from very high loss (60 dB/cm) to low-loss regime (~5 dB/cm) is reported for a 220 nm x 500 nm Si(3)N(4) wire at 900 nm wavelength. This opens prospects to implement very low loss waveguides.

Generation of human-induced pluripotent stem cell line from PBMC of healthy donor using integration-free Sendai virus technology.

We developed a well-characterized human induced pluripotent stem cell (iPSC) line obtained from healthy individuals' peripheral blood mononuclear cells (PBMC). The PBMCs were primed and reprogrammed using a non-integrating sendai viral vector, and the iPSC lines demonstrated complete differentiation capacity. This line, YBLi004-A, is available and registered in the human pluripotent stem cell registry. The line's legitimacy was validated using pluripotent marker expression, in vitro differentiation into three germ layers (ectoderm, mesoderm, and endoderm), karyotyping, and STR analysis. This iPSC line could be used as a healthy control for studies involving disease-specific-iPSCs, e.g. drug toxicity and efficacy testing.

Direct cloning of double-stranded RNAs from RNase protection analysis reveals processing patterns of C/D box snoRNAs and provides evidence for widespread antisense transcript expression.

We describe a new method that allows cloning of double-stranded RNAs (dsRNAs) that are generated in RNase protection experiments. We demonstrate that the mouse C/D box snoRNA MBII-85 (SNORD116) is processed into at least five shorter RNAs using processing sites near known functional elements of C/D box snoRNAs. Surprisingly, the majority of cloned RNAs from RNase protection experiments were derived from endogenous cellular RNA, indicating widespread antisense expression. The cloned dsRNAs could be mapped to genome areas that show RNA expression on both DNA strands and partially overlapped with experimentally determined argonaute-binding sites. The data suggest a conserved processing pattern for some C/D box snoRNAs and abundant expression of longer, non-coding RNAs in the cell that can potentially form dsRNAs.

Toward Personalized Medicine Approaches for Parkinson Disease Using Digital Technologies.

Parkinson disease (PD) is a complex neurodegenerative disorder that afflicts over 10 million people worldwide, resulting in debilitating motor and cognitive impairment. In the United States alone (with approximately 1 million cases), the economic burden for treating and caring for persons with PD exceeds US $50 billion and myriad therapeutic approaches are under development, including both symptomatic- and disease-modifying agents. The challenges presented in addressing PD are compounded by observations that numerous, statistically distinct patient phenotypes present with a wide variety of motor and nonmotor symptomatic profiles, varying responses to current standard-of-care symptom-alleviating medications (L-DOPA and dopaminergic agonists), and different disease trajectories. The existence of these differing phenotypes highlights the opportunities in personalized approaches to symptom management and disease control. The prodromal period of PD can span across several decades, allowing the potential to leverage the unique array of composite symptoms presented to trigger early interventions. This may be especially beneficial as disease progression in PD (alongside Alzheimer disease and Huntington disease) may be influenced by biological processes such as oxidative stress, offering the potential for individual lifestyle factors to be tailored to delay disease onset. In this viewpoint, we offer potential scenarios where emerging diagnostic and monitoring strategies might be tailored to the individual patient under the tenets of P4 medicine (predict, prevent, personalize, and participate). These approaches may be especially relevant as the causative factors and biochemical pathways responsible for the observed neurodegeneration in patients with PD remain areas of fluid debate. The numerous observational patient cohorts established globally offer an excellent opportunity to test and refine approaches to detect, characterize, control, modify the course, and ultimately stop progression of this debilitating disease. Such approaches may also help development of parallel interventive strategies in other diseases such as Alzheimer disease and Huntington disease, which share common traits and etiologies with PD. In this overview, we highlight near-term opportunities to apply P4 medicine principles for patients with PD and introduce the concept of composite orthogonal patient monitoring.

The snoRNA MBII-52 (SNORD 115) is processed into smaller RNAs and regulates alternative splicing.

The loss of HBII-52 and related C/D box small nucleolar RNA (snoRNA) expression units have been implicated as a cause for the Prader-Willi syndrome (PWS). We recently found that the C/D box snoRNA HBII-52 changes the alternative splicing of the serotonin receptor 2C pre-mRNA, which is different from the traditional C/D box snoRNA function in non-mRNA methylation. Using bioinformatic predictions and experimental verification, we identified five pre-mRNAs (DPM2, TAF1, RALGPS1, PBRM1 and CRHR1) containing alternative exons that are regulated by MBII-52, the mouse homolog of HBII-52. Analysis of a single member of the MBII-52 cluster of snoRNAs by RNase protection and northern blot analysis shows that the MBII-52 expressing unit generates shorter RNAs that originate from the full-length MBII-52 snoRNA through additional processing steps. These novel RNAs associate with hnRNPs and not with proteins associated with canonical C/D box snoRNAs. Our data indicate that not a traditional C/D box snoRNA MBII-52, but a processed version lacking the snoRNA stem is the predominant MBII-52 RNA missing in PWS. This processed snoRNA functions in alternative splice-site selection. Its substitution could be a therapeutic principle for PWS.

One pot synthesis and anticancer activity of dimeric phloroglucinols.

A series of dimeric phloroglucinol compounds were synthesized in a single step using commercially available phloroglucinol and methanesulfonic acid. Based on the reported anticancer activity of plant derived dimeric phloroglucinols, these synthesized compounds were evaluated for their in vitro anti-proliferative activities against various cancer cell lines. Several compounds demonstrated in vitro cytotoxic effects across a wide array of tumor cell types. The compound 29 with pyridin-3-yl group on linker methylene and two diisovaleryl phloroglucinol moieties was found to be the most active in all the five cancer cell lines having a low IC(50) of 5.5 µM in colon cancer cell lines (HCT116).

Should all endoscopically excised rectal polyps be tattooed? A plea for localization.

BACKGROUND: More than 5-8 % of endoscopically removed rectal polyps presumed to be benign contain invasive carcinoma. Tattooing has been advocated for follow-up localization of the resection site. Despite proven benefits, the authors propose that tattooing is not routinely performed when benign-appearing rectal polyps are endoscopically excised, thereby confounding management when invasive cancer is found. The secondary goal of the study was to determine the frequency of localization, polyp characteristics, and accuracy of predicting malignant potential at the authors' institution. METHODS: All patients with rectal neoplasia discovered during endoscopic polypectomy from 1 January 2003 to 1 August 2010 were retrospectively identified from Temple University Hospital's Tumor Registry. Demographic and clinical data were extracted from medical records including polyp size, gross appearance, pathology, resection margins, location based on preoperative colonoscopy, initial removal technique, tattoo performance, and ensuing procedures. RESULTS: During the study period, 49 patients had colonoscopic excision of presumed benign rectal polyps with ensuing diagnosis of neoplasia in the specimen. The malignant histology included adenocarcinoma (n = 5), carcinoma in situ (n = 21), carcinoid (n = 22), and composite carcinoid (n = 1). Only two polyps were tattooed at the initial polypectomy. Three polyps were "suspicious for malignancy." None of the suspicious polyps were tattooed. One of the suspicious lesions was an adenocarcinoma, and the remaining two were benign. The distance from the anal verge was noted in only seven patients. The predominant excision technique was hot snare polypectomy (n = 29). None of the incomplete polyp excisions for 15 patients were "suspicious for malignancy" or tattooed. Several strategies were used to manage incomplete resections including surveillance (40 %), repeat colonoscopic polypectomy (27 %), and surgery (33 %). CONCLUSIONS: Most malignant rectal polyps are neither diagnosed nor tattooed at initial colonoscopy. Moreover, the distance of the polyp from the anal verge is rarely measured, and gross characteristics are not well described. Tattooing of all endoscopically excised rectal polypectomy sites would avoid confounding of subsequent identification and management.

Anticancer activity of sclerotiorin, isolated from an endophytic fungus Cephalotheca faveolata Yaguchi, Nishim. & Udagawa.

Biodiversity provides critical support for drug discovery. A significant proportion of drugs are derived, directly or indirectly, from biological sources. Through high throughput screening (HTS) and bioassay-guided isolation, bioactive compound sclerotiorin has been isolated from an endophytic fungus Cephalotheca faveolata. Sclerotiorin was found to be potent anti-proliferative against different cancer cells. In this study sclerotiorin has been found to induce apoptosis in colon cancer (HCT-116) cells through the activation of BAX, and down-regulation of BCL-2, those further activated cleaved caspase-3 causing apoptosis of cancer cells.

Physician centricity in the deployment of digital solutions for neurological conditions.

The widespread deployment of telemedical approaches to managed care during the CoV2 pandemic has provided an opportunity for clinicians to engage in the development and refinement of this mode of delivery. This also represents a pivotal moment to help effect a paradigm shift in how new and more sophisticated digital health services are designed and delivered with the caregiver playing a guiding role. Building on momentum this way can allow the fuller potential of digital health to be realized by focusing on "end user pull" which balances the omnipresent "technology push" of the consumer product and medical device industries. Perhaps nowhere is this more critical than in the care of neurological illnesses where patient-provider interactions must be managed frequently and rely on a complex battery of data measures. The emergent role of the physician-entrepreneur can be envisioned, complimenting established physician-scientist career paths and represents a timely and opportune moment to refine medical education curricula.

Ets-1 expression and gemcitabine chemoresistance in pancreatic cancer cells.

Gemcitabine, a novel pyrimidine nucleoside analog, has become the standard chemotherapeutic agent for pancreatic cancer patients. The clinical impact of gemcitabine remains modest owing to the high degree of inherent and acquired resistance. There are various lines of evidence that confirm the role of Ets-1, a proto-oncoprotein, in tumor invasion, progression, and chemoresistance. This study examines a hypothesis that implicates Ets-1 in the development of gemcitabine-resistance in pancreatic cancer cells. Ets-1 protein expression was assessed in parental pancreatic cancer cells and their gemcitabine-resistant clones. Western blot analysis revealed elevated levels of Ets-1 protein expression in gemcitabine-resistant PANC1(GemRes) (4.8-fold increase; P < 0.05), MIA PaCa2(GemRes) (3.2-fold increase; P < 0.05), and Capan2(GemRes) (2.1-fold increase; P < 0.05) cells as compared to their parental counterparts. A time course analysis was conducted to determine the change in Ets-1 expression in the parental cells after incubation with gemcitabine. Reverse transcriptase quantitative real-time PCR (RT-qPCR) and Western blot analysis revealed a significant increase in Ets-1 expression. All the three parental cells incubated with gemcitabine showed elevated Ets-1 protein expression at 6 h. By 24 h, the expression level had decreased. Using small interfering RNA (siRNA) against Ets-1 in gemcitabine-resistant cells, we demonstrated a reversal in gemcitabine chemosensitivity and also detected a marked reduction in the expression of the Ets-1 target genes MMP1 and uPA. Our novel finding demonstrates the significance of Ets-1 in the development of gemcitabine chemoresistance in pancreatic cancer cells. Based on these results, a new siRNA-based therapeutic strategy targeting the Ets-1 genes can be designed to overcome chemoresistance.

Envisioning Post-pandemic Digital Neurological, Psychiatric and Mental Health Care.

The SARS-Cov-2 pandemic placed a dramatic burden on managed healthcare and perhaps nowhere as evident as in neurological and psychiatric disease care. This said, the duration of the pandemic mandated adaptability of the entire care system and the oft-vaunted benefits of telehealth and telemedicine were subjected to deep scrutiny at scale. Positive experiences were reported by both patients and providers from routine check-ups, to use of cognitive behavioral therapy associated with mental disorders, and management of complex diseases such as multiple sclerosis and other neurological and psychiatric conditions. Integration into standard care looks likely in the post pandemic era with many healthcare systems moving to expand reimbursement categories and develop equitable incentive models for developers and providers. In this commentary we share perspective on how the future of care may evolve through hybrid delivery models, and the advent of new therapeutic approaches which can address pain points identified during the pandemic.

Trimethylamine modulates dauer formation, neurodegeneration, and lifespan through tyra-3/daf-11 signaling in Caenorhabditis elegans.

In the nematode Caenorhabditis elegans, signals derived from bacteria in the diet, the animal's major nutrient source, can modulate both behavior and healthspan. Here we describe a dual role for trimethylamine (TMA), a human gut flora metabolite, which acts as a nutrient signal and a neurotoxin. TMA and its associated metabolites are produced by the human gut microbiome and have been suggested to serve as risk biomarkers for diabetes and cardiovascular diseases. We demonstrate that the tyramine receptor TYRA-3, a conserved G protein-coupled receptor (GPCR), is required to sense TMA and mediate its responses. TMA activates guanylyl cyclase DAF-11 signaling through TYRA-3 in amphid neurons (ASK) and ciliated neurons (BAG) to mediate food-sensing behavior. Bacterial mutants deficient in TMA production enhance dauer formation, extend lifespan, and are less preferred as a food source. Increased levels of TMA lead to neural damage in models of Parkinson's disease and shorten lifespan. Our results reveal conserved signaling pathways modulated by TMA in C. elegans that are likely to be relevant for its effects in mammalian systems.

Regulation of alternative splicing by short non-coding nuclear RNAs.

Recent results from deep-sequencing and tiling array studies indicated the existence of a large number of short, metabolically stable, non-coding RNAs. Some of these short RNAs derive from known RNA classes like snoRNA or tRNAs. There are intriguing similarities between short non-coding nuclear RNAs and oligonucleotides used to change alternative splicing events, which usually target a disease-relevant RNA. We review the current knowledge of this emerging class of RNAs and discuss evidence that some of these short RNAs could function in alternative splice site selection.

Polarization properties of two-dimensional slot waveguides.

We propose and study slot waveguide geometries where both quasi-TE and quasi-TM modes may propagate highly confined within the same low-index slot region. Conventional horizontal and vertical slot waveguides can only provide high slot confinement for either the quasi-TM or quasi-TE modes, respectively. Different two-dimensional slot waveguide structures are analyzed in terms of their mode characteristics, such as the effective index, the confinement factor, and the overlap of quasi-TE and -TM modes within the slot. Attention is also paid to practical manufacturability. Various waveguide structures can be tailored to have zero birefringence or equal confinement at both polarizations. Values for the confinement factors and the overlap of the two polarizations, in the slot region, can reach 0.4 to 0.5.

Control of optical mode properties in cross-slot waveguides.

A cross-slot waveguide geometry provides high confinement of the mode field of both fundamental quasi-TE and quasi-TM modes in geometrically perpendicular slots. A unique possibility to tailor optical mode characteristics such as effective index and confinement factor quite independently for the two polarizations with geometric and material parameters is shown. Nonbirefringent cross-slot geometries are presented. Fabrication related tolerances of the cross-slot geometry for low birefringence operation are studied. Means to externally tune the birefringence by a thermo-optic effect is also analyzed. Fabrication of a cross-slot waveguide test structure is demonstrated.

Anti-inflammatory and anticancer activity of ergoflavin isolated from an endophytic fungus.

Biodiversity is a major resource for identification of new molecules with specific therapeutic activities. To identify such an active resource, high throughput screening (HTS) of the extracts prepared from such diversity are examined on specific functional assays. Based on such HTS studies and bioactivity-based fractionation, we have isolated ergoflavin, a pigment from an endophytic fungus, growing on the leaves of an Indian medicinal plant Mimosops elengi (bakul). We report here the isolation, structure elucidation, and biological properties of this compound, which showed good anti-inflammatory and anticancer activities.

Rapid generation of splicing reporters with pSpliceExpress.

Almost all human protein-coding transcripts undergo pre-mRNA splicing and a majority of them is alternatively spliced. The most common technique used to analyze the regulation of an alternative exon is through reporter minigene constructs. However, their construction is time-consuming and is often complicated by the limited availability of appropriate restriction sites. Here, we report a fast and simple recombination-based method to generate splicing reporter genes, using a new vector, pSpliceExpress. The system allows generation of minigenes within one week. Minigenes generated with pSpliceExpress show the same regulation as displayed by conventionally cloned reporter constructs and provide an alternate avenue to study splice site selection in vivo.