Therapeutic Demands Research Articles

Rapid, serial, non-invasive quantification of Pseudomonas aeruginosa in live mice with a selectable marker-free autoluminescent strain

Pseudomonas aeruginosa is an increasingly prevalent pathogen that has become a serious health concern due to an increasing incidence of multidrug-resistant (MDR) hospital-acquired infections. The emergence of MDR-P. aeruginosa coupled with shrinking antibiotic pipelines has increased the demand for new antimicrobials and therapeutics. An effective tool for drug screening both in vitro and in vivo can facilitate the discovery of drugs and regimens for treating P. aeruginosa infection. Here, for the first time, we combined the mini-Tn7 system and Xer/dif recombinase system to construct a stable and selectable marker-free autoluminescent P. aeruginosa (SfAlPa) by one step. Afterwards, in vitro and in vivo activities of several antibiotics including amikacin, biapenem, levofloxacin and polymyxin B were assessed using SfAlPa. This study demonstrated that the use of SfAlPa could significantly facilitate rapid real-time evaluating the activities of compounds. Compared to prevailing methods, this method reduces the time, effort, animals and costs consumed in the discovery of new drugs against P. aeruginosa. Additionally, the methodology described in this study could be easily modified for construction of selectable marker-free reporter strain in other Gram-negative bacteria.

The Brave New World of Manufacturing Speed, Capacity, and Quality

Genetic Engineering & Biotechnology NewsVol. 40, No. 6 Sponsored ContentFree AccessThe Brave New World of Manufacturing Speed, Capacity, and QualityPublished Online:11 Jun 2020https://doi.org/10.1089/gen.40.06.12AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail The biopharmaceutical industry is under pressure to increase the speed and capacity of manufacturing, without sacrificing quality. Growth in demand for novel therapeutics continues to strain existing capacity and drives the need for new strategies to accelerate bioprocessing.The COVID-19 pandemic is significantly amplifying this pressure and pushing industry to rethink conventional development and manufacturing strategies. A recent New England Journal of Medicine1 article highlighted this new paradigm, as hundreds of companies race to develop therapeutics and vaccines at “pandemic speed”:Manufacturers face growing pressure to streamline their processes and run more activities in parallel.Manufacturing development, scale-up, clinical lots, commercial scale production, and validation of processes are occurring much earlier in the pharmaceutical value chain to support clinical development.Manufacturers are investing in increased capacity “at-risk,” to prepare for vaccine manufacture, without knowing whether a candidate will be safe and effective.The challenges facing industry extend beyond simply increasing speed and compressing timelines: It is also a matter of scale. Pandemic-related therapeutics and vaccines will be produced in the billions of doses—not millions—but the massively larger volumes will need to be manufactured in the context of complex social distancing guidelines designed to keep personnel safe.Quality Control at a CrossroadsInnovative approaches to quality control (QC) that are faster and highly automated and deliver the accuracy needed to support confident decision-making at a time of increased risk will support these goals. Current QC processes used to detect microbial contamination are simply too time- and labor-intensive. New technologies are needed to take QC processes to the next level of speed and sophistication to enable manufacturers of biopharmaceuticals and vaccines to meet regulatory requirements, sustain competitive advantage, and, ultimately, serve patient needs and safety.A critical cornerstone of QC in drug manufacturing is the ability to monitor and detect microbial contamination. Rapid Micro Biosystems has developed an automated, high-throughput platform, Growth Direct™, that replaces slow, error-prone, manual QC processes, allowing manufacturers to confidently accelerate bioprocessing, maximize capacity, and reduce operational risk and downtime, all while maintaining the highest standards of data integrity.The platform supports even those bio-processes where speed and capacity are at a premium, such as preparing for COVID-19 vaccine manufacturing. Importantly, this high-throughput platform can be operated with minimal human interaction. Remote monitoring capabilities further maximize efficiency and minimize the number of lab employees required to run the tests, enabling social distancing.Global Adoption of New QC TechnologyDemand for automated QC technology is growing rapidly, particularly among manufacturers of biologics, sterile injectables, vaccines, and cell and gene therapies. To date, the Growth Direct platform for microbial detection has been adopted by the majority of the top 20 biopharmaceutical companies and is used by industry leaders in their commercial manufacture of groundbreaking therapeutics, including complex biologics and cell and gene therapies.There is global interest in investing in technologies that advance a new biopharmaceutical manufacturing paradigm. Rapid Micro Biosystems recently closed a round of financing which will enable the company to accelerate commercial expansion of the Growth Direct platform, and expand global supply chains and manufacturing networks to meet growing demand. The investment will also allow the company to bring new complimentary products to market, including those to accelerate response to pandemics.Motortion/iStockA New Normal at Pandemic SpeedWhen the pandemic subsides and new treatments and vaccines are widely available, there will be no going back to conventional methods of manufacturing. Gains in speed, capacity, and agility of workflows and infrastructure will become the new normal. A key element of this paradigm shift will be the integration of automated, high-throughput approaches to QC. This cornerstone technology paves the way for increases in speed, capacity, and patient safety, while reducing business risk and ensuring critical data integrity, setting the stage for the brave new world of manufacturing.For more information about us visit rapidmicrobio.com

Key Challenges in Designing CHO Chassis Platforms

Following the success of and the high demand for recombinant protein-based therapeutics during the last 25 years, the pharmaceutical industry has invested significantly in the development of novel treatments based on biologics. Mammalian cells are the major production systems for these complex biopharmaceuticals, with Chinese hamster ovary (CHO) cell lines as the most important players. Over the years, various engineering strategies and modeling approaches have been used to improve microbial production platforms, such as bacteria and yeasts, as well as to create pre-optimized chassis host strains. However, the complexity of mammalian cells curtailed the optimization of these host cells by metabolic engineering. Most of the improvements of titer and productivity were achieved by media optimization and large-scale screening of producer clones. The advances made in recent years now open the door to again consider the potential application of systems biology approaches and metabolic engineering also to CHO. The availability of a reference genome sequence, genome-scale metabolic models and the growing number of various “omics” datasets can help overcome the complexity of CHO cells and support design strategies to boost their production performance. Modular design approaches applied to engineer industrially relevant cell lines have evolved to reduce the time and effort needed for the generation of new producer cells and to allow the achievement of desired product titers and quality. Nevertheless, important steps to enable the design of a chassis platform similar to those in use in the microbial world are still missing. In this review, we highlight the importance of mammalian cellular platforms for the production of biopharmaceuticals and compare them to microbial platforms, with an emphasis on describing novel approaches and discussing still open questions that need to be resolved to reach the objective of designing enhanced modular chassis CHO cell lines.

COVID-19: an update on diagnostic and therapeutic approaches

The unexpected pandemic set off by the novel coronavirus 2019 (COVID-19) has caused severe panic among people worldwide. COVID-19 has created havoc, and scientists and physicians are urged to test the efficiency and safety of drugs used to treat this disease. In such a pandemic situation, various steps have been taken by the government to control and prevent the Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2). This pandemic situation has forced scientists to rework strategies to combat infectious diseases through drugs, treatment, and control measures. COVID-19 treatment requires both limiting viral multiplication and neutralizing tissue damage induced by an inappropriate immune reaction. Currently, various diagnostic kits to test for COVID-19 are available, and repurposing therapeutics for COVID-19 has shown to be clinically effective. As the global demand for diagnostics and therapeutics continues to rise, it is essential to rapidly develop various algorithms to successfully identify and contain the virus. This review discusses the updates on specimens/samples, recent efficient diagnostics, and therapeutic approaches to control the disease and repurposed drugs mainly focusing on chloroquine/hydroxychloroquine and convalescent plasma (CP). More research is required for further understanding of the influence of diagnostics and therapeutic approaches to develop vaccines and drugs for COVID-19.

Nanoneedle Platforms: The Many Ways to Pierce the Cell Membrane

AbstractEstablishing techniques to efficiently and nondestructively access the intracellular milieu is essential for many biomedical and scientific applications, ranging from drug delivery, to electrical recording, to biochemical detection. Cell penetration using nanoneedle arrays is currently a research focus area because it not only meets the increasing therapeutic demands of cell modifications and genome editing, but also provides an ideal platform for tracking long‐term intracellular information. Although the precise mechanism driving membrane penetration by nanoneedle arrays is still unclear, the low cytotoxicity, wide range of delivered materials, diverse cell type targets, and simple material structures of nanoneedle arrays make these splendid platforms for cell access. Here, the recent progress in this field is reviewed by examining device architectures and discussing mechanisms for nanoneedle penetration, and the major studies demonstrating the most general applicability of nanoneedle arrays, typical methodologies to access the intracellular environment using nanoneedles with spontaneous or assisted penetration modes, as well as biosafety aspects are presented. This review should be valuable for deeply understanding the materials fabrication principles, device designs, cell penetration methodologies, biosafety aspects, and application strategies of nanoneedle array‐based systems that are of crucial importance for the development of future practical biomedical platforms.

Design, synthesis and biological evaluation of novel pyrazole sulfonamide derivatives as dual COX-2/5-LOX inhibitors

The current therapeutic demand focuses more on the discovery of safer NSAIDs rather than exploring more potent alternatives. The dual COX-2/5-LOX inhibition is a promising strategy for designing compounds with an enhanced efficacy, reduced side-effects and a broader anti-inflammatory spectrum in comparison to classical NSAIDs. In the present study, a hybridization strategy was adopted to combine the binding features of the non-selective COX inhibitor “sulindac” and the selective COX-2 inhibitor “celecoxib” which show 5-LOX inhibitory activity with that of licofelone and a celecoxib pyridone analogue which show dual COX-2/5-LOX inhibitory activity to design new series of pyrazole sulfonamide derivatives which, by design, should possess dual COX-2/5-LOX inhibitory activity. All the newly synthesized compounds were initially tested for their potential analgesic activity, then candidates that showed potential analgesic activity, were selected for the subsequent anti-inflammatory activity evaluation, as well as, ulcerogenicity testing. Moreover, in vitro assessment of their COX-1, COX-2 and 5-LOX inhibitory activities were performed. The benzothiophen-2-yl pyrazole carboxylic acid derivative 5b showed the most potent analgesic and anti-inflammatory activities surpassing that of celecoxib and indomethacin. It showed potent COX-1, COX-2 and 5-LOX inhibitory activity with IC50 of 5.40, 0.01 and 1.78 μM, respectively, showing a selectivity index of 344.56 that was much better than the used reference standards and its parent compounds, confirming its selectivity towards COX-2 over COX-1. The prodrug ester derivatives 6c and 6d showed equipotent activity to their parent compound 5b with no gastric ulcerogenicity. Molecular docking simulations confirmed that the newly synthesized compounds possess the structural features required for binding to the target enzymes COX-2 and 5-LOX.

P432 Effects and safety of a colon-long absorbing budesonide product in patients with mild to moderate ulcerative colitis

Abstract Background Budesonide is an oral corticosteroid, which is widely used in moderate-to-severe ulcerative colitis (UC) to obtain and maintain remission in those cases, where 5-aminosalicylic-acid was ineffective. Unlike previous forms of budesonide absorbing from the ileal and ascending colon region, the new-generation budesonide-MMX contains a formula, that allows absorption throughout the whole colon, and therefore in the whole potentially inflamed area in UC. We aimed to evaluate the effects and safety of budesonide-MMX in our UC patients who meet the above mentioned therapeutic demands in a real-life study. Methods We enrolled 22 patients with mild to severe UC in this single-centre prospective study until August of 2019. Patients received 9 mg oral budesonide-MMX once daily until 8 weeks. Laboratory parameters (cholesterol, triglyceride, CRP) and serum hormone levels (parathormone [PTH], dehydroepiandrosterone [DHEA] and cortisol) were monitored before and after the 8-week therapy to follow metabolic and hormonal changes. During these visits, body composition analysis was also performed with InBody 770 machine to observe the adverse steroid effects of budesonide-MMX in respect of body fat mass, body mass index, protein content of the body and bone mineral content. Disease activity was followed by the partial Mayo (pMayo) score. Statistical analysis was performed by paired t-test and Wilcoxon signed-rank test with SigmaPlot 1.25. Results The total of 22 patients (age: 44.4 ± 15.1 years, 6 male and 16 female patients) had received the 2-month budesonide-MMX therapy (2.0 ± 0.3 months). Mean disease duration was 8.3 years. By the end of follow-up, 15 (68.2%) patients experienced remission and 7 patients (31.8%) were primary non-responders. The disease activity decreased significantly from the mean of 3.95 to 1.64 (p < 0.001). No significant changes were observable in case of any body composition analysis parameter. Regarding the laboratory parameters, serum cholesterin level showed a significant increase (p < 0.001), while triglyceride and CRP showed did not show significant changes. Serum cortisol levels were elevated (p < 0.001), while PTH and DHEA showed no significant decrease. Only two patients experienced side effects: one of them hypertonia, headache and acnes, while the other patient experienced mild diarrhoea. One patient had a relapse during the treatment. Conclusion In our study, budesonide-MMX proved to be safe by bringing up a low number of side effects, while more than two-thirds of the patients could reach remission with this short-term therapy. Hormonal changes were not mentionable, although the drug’s effects on serum lipid content have to be examined further.

Management of Dental Emergencies in Children and Adolescents

A unique, multidisciplinary manual for the treatment of pediatric dental emergencies for general practitioners and non-pediatric specialists. Management of Dental Emergencies in Children and Adolescents presents the diagnostic skills, treatment options, and management strategies necessary to provide effective and appropriate dental care for children and adolescents. This authoritative manual helps dental practitioners manage potentially stressful situations with children and adolescents while improving their competence in a wide range of urgent pediatric situations. An emphasis on managing the therapeutic demands of both younger patients and their parents enables readers to have greater confidence in handling demanding emergency situations in daily practice. An expert team of contributors explain how to manage tooth substance loss, endodontic problems in deciduous teeth, the long‐term consequences of early tooth loss, the dental issues related to oral health, and more. Guiding practitioners through the unique challenges of pediatric dental emergencies, this book: - Explains the differences in treating and managing dental emergencies in children compared to adults - Covers all types of pediatric dental emergencies including open pulp in permanent and deciduous teeth, missing teeth, and non-infective dental conditions - Offers clinical vignettes and photographs to highlight clinical relevance - Includes chapters by experts in multiple disciplines such as endodontics, restorative dentistry, pediatric dentistry, prosthodontics, and orthodontics The first textbook to focus exclusively on young patients in need of acute dental care, Management of Dental Emergencies in Children and Adolescents is a much-needed resource for general and specialist dentists as well as trainee and specialist pediatric dentists.

Application of medicinal plants in several dermatovenerological entities.

Treatment of skin conditions with medicinal plants has been an ongoing human activity lasting over thousands of years. The use of specific plant species developed regionally, based on local flora. Commonly used medicinal plants for dermatological complaints are: Phlebodium aureum (L.) J. Sm., Ginkgo biloba L., Rosmarinus officinalis L., Panax ginseng C.A.Mey., Allium cepa L., Aloe vera (L.) Burm.f., Capsicum annuum L., Berbe ris aquifolium Pursh, Camellia sinensis (L.) Kuntze, and Podophyllum peltatum L. The demand for complementary therapeutics is an emerging trend due to the awareness of potential side effects that synthetic drugs might cause. More scientific evidence and better documentation are needed before advising dermatologic patients on herbal medicinal treatment. Standardised extracts and formulations with proven clinical efficacy should be developed for this cause. Here provided review entails the use of herbal medicinal products in the treatment of frequent chronic skin diseases, such as vitiligo, alopecia, psoriasis and genital warts.

The Efficacy and Safety of Myelophil, an Ethanol Extract Mixture of Astragali Radix and Salviae Radix, for Chronic Fatigue Syndrome: A Randomized Clinical Trial

Background: There is a strong demand for therapeutics to treat chronic fatigue syndrome (CFS), although there are limitations. Myelophil, which is a combination of extracts from Astragali Radix and Salviae Miltiorrhizae Radix, has been clinically used to treat fatigue-related disorders in South Korea. We conducted a randomized controlled clinical trial of Myelophil in patients with CFS and evaluated its efficacy and safety in two hospitals. Methods: We enrolled 98 participants (M: 38, F: 60) with CFS in a phase 2 trial of oral Myelophil (2 g daily) or placebo for 12 weeks. The primary end point was a change in the Chalder fatigue scale, as scored by a numeric rating scale (NRS). The secondary end points included changes in the visual analogue scale, fatigue severity scale (FSS), and 36-item short-form health survey (SF-36). Biomarkers of oxidative stress and cytokines were evaluated by blood tests. Results: Ninety-seven participants (48 in the Myelophil group and 49 in the placebo group) completed the trial. An analysis of all participants showed that Myelophil slightly improved fatigue symptoms compared with those of the placebo, but this effect was not statistically significant (p > 0.05 for the NRS, VAS, FSS, and SF-36). By contrast, an analysis of the subpopulation (53 participants, M: 24, F: 29) with severe symptoms (≥63, median NRS value of total participants) showed a statistically significant improvement in fatigue symptoms in the Myelophil group compared with the placebo (p < 0.05 for NRS, FSS, and SF-36). There were no significant changes in the biomarkers for oxidative stress and cytokines before or after the treatment. No Myelophil-related adverse response was observed during the trial. Conclusion: These results support the hypothesis that Myelophil can be a therapeutic candidate to manage CFS and provide the rationale for its progression to a phase 3 clinical trial. Clinical Trial Registration: www.ClinicalTrials.gov, identifier KCT0002317.

Preclinical CAR-T cell target safety, biodistribution, and tumor infiltration analysis using in situ hybridization technology.

112 Background: Chimeric antigen receptor (CAR) T cell therapy is highly effective in treating hematologic malignancies, and major efforts are being made to achieve similar efficacy in solid tumors. The greater potency of CAR-T cells compared to antibody therapeutics demands a more stringent CAR-T target safety assessment to avoid adverse events resulting from “on-target/off-tumor” activity. Furthermore, it is critical to track and monitor CAR+ T cells within intact tissue and tumor to understand the mechanisms underlying off-tumor toxicity and efficacy in tumor killing. Methods: We employed the RNAscope in situ hybridization (ISH) technology to assess target expression specificity and to track CAR-T cell distribution and activation in xenograft and host tissues using the RPMI-8226 xenograft mouse model. Results: For the CAR-T target candidates, Target X and Target Y, RNA ISH revealed that Target X was only expressed in the xenograft tumor and in no mouse organs, while Target Y was found to be expressed at low levels in mouse lung and liver, as well as in the xenograft tumor. Duplex RNA ISH assays with probes targeting the CAR 3’ UTR and either IFNG or GZMB allowed for highly sensitive and specific detection of CAR-T cells and their activation state in both tumor and normal tissues from vehicle, Target X CAR-T cell, or Target Y CAR-T cell treated mice. Activated Target X CAR-T cells expressing GZMB and IFNG were found only in the xenograft tumor, where Target X was expressed. In contrast, activated Target Y CAR-T cells were found almost exclusively in mouse lung and liver, with very few Target Y CAR-T cells being found in the xenograft tumor. Lastly, a multiplex ISH-IHC approach confirmed the presence of activated Target X CAR-T cells in the xenograft tumor through simultaneous detection of the Target X CAR 3’ UTR, IFNG, GZMB, and CD3. Conclusions: These data demonstrate how the RNAscope assay can be utilized for CAR-T cell efficacy and safety/toxicity assessment in preclinical models by detecting very low levels of target antigen expression in off-tumor tissues and monitoring CAR-T cell pharmacodynamics and activation in tumor models and can also be applied for assessing TCR-T cell activity in tumors.

Combating tigecycline resistant Acinetobacter baumannii: A leap forward towards multi-epitope based vaccine discovery

Global emergence of Tigecycline resistant Acinetobacter baumannii (TRAB) is on the horizon and poses a very serious threat to human health. There is a pressing demand for suitable therapeutics against this pathogen, particularly a vaccine to protect against TRAB infections. We present a comprehensive investigation of the complete proteome of a TRAB AB031 strain to predict promiscuous antigenic, non-allergenic, virulent B-cell derived T-cell epitopes and formulate a multi-epitope vaccine against the pathogen. We identified epitopes from three proteins: outer membrane protein assembly factor (BamA), fimbrial biogenesis outer membrane usher protein (FimD) and type IV secretion protein (Rhs) that are appropriate for vaccine design. These proteins constitute the core proteome of the pathogen, are essential, localized at the pathogen surface, non-homologous to humans, mice and to the beneficial probiotic bacteria that reside the human gut. Moreover, these proteins are ideal candidates for experimental investigation as they have favorable physicochemical properties and have strong cellular interacting networks. The predicted epitopes: FPLNDKPGD (BamA), FVHAEEAAA (FimD) and YVVAGTAAA (Rhs) have exo-membrane topology for efficient recognition of the host immune system and high affinity for the most prevalent allele in human populations, the DRB*0101. These epitopes were linked and attached to an adjuvant to enhance its antigenicity. The multi-epitope vaccine-construct was docked with the TLR4 receptor to assess its affinity for the protein and thus its presentation to the host immune system. Docking results were validated through molecular dynamics simulations and binding free energies were calculated using the molecular mechanics/generalized Born (MM-GBSA) method. In conclusion, we expect the designed chimeric vaccine is highly likely to be effective against infections caused by TRAB.

STROKE MIMICS: A PSYCHOGENIC STROKE PATIENT TREATED WITH ALTEPLASE

For rtPA treatment to be effective it should be initiated within the first 4.5 hours following the onset of a stroke. Such a short therapeutic window demands a rapid diagnosis and decision making on the part of the physician. There are patients with stroke-like symptoms and an initial diagnosis of a stroke, but who are finally diagnosed as suffering from another condition. According to the subject literature, stroke mimics are diagnosed in about 1.4 – 3.5% of patients initially diagnosed as having had an ischemic stroke. Psychogenic strokes (conversion disorders) may be found in as many as 8.2% of stroke patients. Proper diagnosis is especially important in patients eligible for thrombolytic treatment when there is usually not enough time to establish the diagnosis of a stroke mimic, especially one of psychogenic origin. A patient with an initial diagnosis of an ischemic stroke who was treated with intravenous alteplase infusion. The previous two ischemic strokes treated in the same manner had been diagnosed one and two years earlier. In all hospitalizations no rtPA treatment complications had been observed. In our patient a proper neuropsychological examination was performed and a conversion disorder diagnosed. We would like to underline the importance of cooperation between the neuropsychologist and neurology physician within clinical practice.

Abstract 84: Idarucizumab in Patients Treated With Dabigatran Suffering Cerebral Ischemia or Intracranial Hemorrhage: A Retrospective Case Series From Germany

Reversal of anticoagulation in patients treated with NOACs is a rare but sometimes urgently needed therapeutic demand. Idarucizumab is a monoclonal antibody fragment with high affinity for dabigatran reversing its anticoagulant effects within minutes. It is approved for patients on dabigatran with life-threatening or uncontrolled bleeding and those requiring emergency intervention. Case reports and smaller case series suggest a benefit for dabigatran-treated patients suffering ischemic stroke to regain eligibility for rt-PA thrombolysis. To provide insights into the clinical use of idarucizumab in patients under effective dabigatran anticoagulation presenting with signs of ischemic stroke or intracranial hemorrhage in clinical routine, we asked all German Departments of Neurology and Neurosurgery to contribute their retrospective data on the use of idarucizumab collected following product launch in January 2016 to June 2018. In 64 responding stroke centers 112 patients presenting with signs of stroke received idarucizumab. 74 patients treated with dabigatran presented with ischemic stroke. In patients receiving rt-PA thrombolysis following idarucizumab, 87% had a benefit from i.v. thrombolysis with a median NIHSS improvement of 6 points. No symptomatic bleeding complications were observed. Nine patients did not improve functionally. Of those, 4 patients deceased during hospital stay (5.4%). Bolus administration of idarucizumab did not delay thrombolysis indicating its value in this emergency situation. A total of 38 patients had intracranial bleeding (SAH n=1, SDH n=12, ICH n=25). Out of 25 patients presenting with intracerebral hemorrhage and treated with idarucizumab, hematoma growth with clinical worsening was documented in three. Outcome was favorable with a median NIHSS improvement of 4 points in 35 patients. Overall mortality was 13%. In conclusion, idarucizumab is a beneficial therapeutic option for patients under dabigatran treatment presenting with ischemic stroke or intracranial hemorrhage in daily German stroke center routine. A prospective registry with controls is ongoing in Germany.

Dynamic combinatorial chemistry as a rapid method for discovering sequence-selective RNA-binding compounds.

The ever-growing number of RNA species that are recognized as having a role in human disease is driving a demand for novel molecular probes and therapeutics. Producing sequence-selective RNA-binding molecules remains a substantial challenge, however. One approach that has been successful in producing molecules with high affinity and specificity for disease-relevant RNAs is the use of dynamic combinatorial chemistry, a fragment-based method in which fragments combine reversibly in the presence of the target. We describe methods for the design, synthesis, and screening of dynamic combinatorial libraries targeting RNA.

Combating colorectal cancer with immunotherapy: Where are we?

Despite incessant advancement in the therapeutic regimens against colorectal cancer (CRC), treatment failure, metastasis and serious side effects associated with the available therapeutic options demand specific targeted therapies that could help in improving the survival rate with significantly less toxicity, fewer untoward effects and improved efficacy. In-depth studies highlighted the potential of CRC to be immune-responsive, thereby opening a new door in the development of strategies to combat CRC. Immunotherapy has attracted a myriad of researchers with a hope that it could potentially increase the efficacy of the treatment regimens in parallel to the reduction of toxicity. Colorectal tumors undergo immune evasion, suppressing patients' immunity and making them susceptible to infections. Therefore, a viable option could be stimulating a patient's own immune system with the help of immune modulators to fight against CRC. This review briefly discusses the immune responsiveness of CRC as well as tumor-associated antigens in CRC, and highlights the current endeavors of the scientific community in the field of immune modulators against CRC that are under development including varied types of vaccines, checkpoint inhibitors and adoptive T-cell therapy.

Toward Electrophysiology-Based Intelligent Adaptive Deep Brain Stimulation for Movement Disorders.

Deep brain stimulation (DBS) represents one of the major clinical breakthroughs in the age of translational neuroscience. In 1987, Benabid and colleagues demonstrated that high-frequency stimulation can mimic the effects of ablative neurosurgery in Parkinson's disease (PD), while offering two key advantages to previous procedures: adjustability and reversibility. Deep brain stimulation is now an established therapeutic approach that robustly alleviates symptoms in patients with movement disorders, such as Parkinson's disease, essential tremor, and dystonia, who present with inadequate or adverse responses to medication. Currently, stimulation electrodes are implanted in specific target regions of the basal ganglia-thalamic circuit and stimulation pulses are delivered chronically. To achieve optimal therapeutic effect, stimulation frequency, amplitude, and pulse width must be adjusted on a patient-specific basis by a movement disorders specialist. The finding that pathological neural activity can be sampled directly from the target region using the DBS electrode has inspired a novel DBS paradigm: closed-loop adaptive DBS (aDBS). The goal of this strategy is to identify pathological and physiologically normal patterns of neuronal activity that can be used to adapt stimulation parameters to the concurrent therapeutic demand. This review will give detailed insight into potential biomarkers and discuss next-generation strategies, implementing advances in artificial intelligence, to further elevate the therapeutic potential of DBS by capitalizing on its modifiable nature. Development of intelligent aDBS, with an ability to deliver highly personalized treatment regimens and to create symptom-specific therapeutic strategies in real-time, could allow for significant further improvements in the quality of life for movement disorders patients with DBS that ultimately could outperform traditional drug treatment.

Is there an appropriate strategy for treating co-morbid irritable bowel syndrome and bladder pain syndrome?

ABSTRACTIntroduction: Two of the most frequent components of chronic pelvic pain syndrome (CPPS) are irritable bowel syndrome (IBS) and bladder pain syndrome (BPS), characterized by considerable overlapping symptoms and pathophysiology. Currently, its management is challenging meaning there is high the demand for novel efficient therapeutics to aid patient care and to tackle the socioeconomic burden of IBS and BPS. As there are presently no sufficient treatment strategies, identifying the mechanisms that result in their main symptoms is the opportunity for developing appropriate therapies.Areas covered: Herein, the authors explore the potential common treatment strategies for co-morbid IBS and BPS and highlight the absolute need for further research of these deliberating clinical entities.Expert opinion: In the future, the authors summise that the discovery of predictive molecular biomarkers combined with clinical phenotypic categorization will likely allow for more definitive differentiation of patients and thus for better treatment options. Furthermore, it has been suggested that effective IBS treatment strategies would be of great value to co-morbid IBS and BPS therapy.

Biosensing Technologies for Therapeutic Drug Monitoring.

Therapeutic drug monitoring (TDM) is the clinical practice of measuring pharmaceutical drug concentrations in patients' biofluids at designated intervals to allow a close and timely control of their dosage. This practice allows for rapid medical intervention in case of toxicity-related issues and/or adjustment of dosage to better fit the therapeutic demand. Currently, TDM is performed in centralized laboratories employing instruments, such as immunoassay analyzers and mass spectrometers that can be run only by trained personnel. However, the time required for the preparation, samples analysis, and data processing, together with the related financial cost, severely affects the application of TDM in medical practices. Therefore, a new generation of analytical tools is necessary to respond to the timely need of drug administration or reduction aiming at effectively treating oncologic patients. State-of-the-Art Technologies for TDM: Technological advances in the field of nanosciences and biosensors offer the unique opportunity to address such issues. The interest for the so-called nanobiosensors is considerably increasing, particularly in drug discovery and clinical chemistry, even though there are only few examples reporting their use for TDM. The techniques employing nanobiosensors are mainly based on electrochemical, optical, and mass detection systems. In this review, we described the most promising methodologies that, in our opinion, will bring TDM towards the next stage of clinical practice in the future.

Poststatin era in atherosclerosis management: lessons from epidemiologic and genetic studies.

Cardiovascular diseases (CVD) are the leading cause of death worldwide with over 17 million deaths every year and represent a major public health challenge. The last decade has seen the emergence of novel antiatherogenic therapies. Despite intensive lipid and blood pressure interventions, the burden of CVD is expected to markedly progress because of the global aging of the population and increasing exposure to detrimental lifestyle-related risk. Epidemiologic and genetic studies helped to better apprehend the biology of atherosclerosis and allowed pharmaceutical innovation and recent translational successes. This includes the development of novel lipid and glucose-lowering therapies and the leverage of anti-inflammatory therapies. Here, we discuss promises and expectations of emerging scientific and pharmaceutical innovations and translational successes to meet the global therapeutic demand.