This document summarizes a round table discussion on automated patch clamp (APC) assay optimization. The key points are:
1) APC instruments like PatchLiner, Port-a-Patch, and SyncroPatch show excellent correlation with manual patch clamp data.
2) APC allows flexibility in experimentation and good seal success rates can be achieved with standard protocols.
3) APC is useful for screening compounds on primary cells and different channel types. While excised patch recordings are not currently possible, APC enables many experimental features like voltage clamp and fast ligand exchange.
Digital RNAseq Technology Introduction: Digital RNAseq Webinar Part 1QIAGEN
QIAseq RNA is a revolutionary turnkey solution for digital gene expression analysis by NGS. From 10 genes to 1000, from one sample to 100, QIAseq RNA delivers precise results on both ION and Illumina sequencing platforms. The data from QIAseq RNA is directly comparable to expression analysis derived from whole transcriptome sequencing or by qRTPCR, only better, cheaper, faster, and more flexible. This webinar will describe the principles of digital expression analysis by NGS, and review the features and benefits of the QIAseq system, options available, and the integrated data analysis package.
Smallest antibody fragment with only ~15 kDa
Recognize novel/hidden epitopes that conventional antibodies cannot
High stability to function and exist within extreme conditions and intracellular environment
https://www.creative-biolabs.com/sdab/one-stop-solution-for-sdab-development.htm
Digital RNAseq Technology Introduction: Digital RNAseq Webinar Part 1QIAGEN
QIAseq RNA is a revolutionary turnkey solution for digital gene expression analysis by NGS. From 10 genes to 1000, from one sample to 100, QIAseq RNA delivers precise results on both ION and Illumina sequencing platforms. The data from QIAseq RNA is directly comparable to expression analysis derived from whole transcriptome sequencing or by qRTPCR, only better, cheaper, faster, and more flexible. This webinar will describe the principles of digital expression analysis by NGS, and review the features and benefits of the QIAseq system, options available, and the integrated data analysis package.
Smallest antibody fragment with only ~15 kDa
Recognize novel/hidden epitopes that conventional antibodies cannot
High stability to function and exist within extreme conditions and intracellular environment
https://www.creative-biolabs.com/sdab/one-stop-solution-for-sdab-development.htm
“N2MO provides cutting-edge ex vivo insect based screening models for predicting ADMET properties of chemical substances in the early drug discovery phase. Insect models represents excellent model systems due to biological active test situation, combined with fast reproducibility with no need for resynthesizing compound and at same time the model produces data that are reflecting mammalian ADMET properties”
Flow Cytometry Training talks - part 1
This forms the first session of the Garvan Flow , Flow Cytometry Training course. this is a 1 1/2 day training course aimed at giving new and experienced researchers a better understanding of cytometry in medical and biological research.
Next generation sequencing (NGS) of circulating tumor DNA (ctDNA) from patient plasma is becoming more widespread in oncology clinical trials. The noninvasive nature of acquiring these samples is particularly important when resection of representative tumor samples is not advised or not possible. However, profiling of ctDNA has challenges to overcome, such as low concentration of ctDNA shed from the tumor and a low signal:noise ratio caused by somatic alterations with less than 1% variant allele fraction. Improving the sensitivity of these assays to detect low allele frequency events with high confidence requires robust sequencing of low input libraries while employing error correction to reduce background noise. To overcome these challenges, we have incorporated unique molecular identifiers (UMIs) into our NGS workflow. Using these novel adapters paired with our proprietary bioinformatics pipeline (AstraZeneca), the number of false positive variants reported for allele fractions less than 0.5% was reduced tenfold. We also refined our curation based on the mapping quality and strand bias in the vicinity of each variant to further reduce the background noise. The use of xGen® Dual Index UMI Adapters—Tech Access (Integrated DNA Technologies) has enabled us to sequence thousands of plasma samples from diverse tumor indications and at differing time points during our trials. The generated data are highly informative with the potential to answer critical questions relating to individual response or resistance to experimental therapies. During this webinar, we discuss our current NGS ctDNA workflow and our future plans to increase our sequencing sensitivity with these novel UMI adapters.
Advances in Mass Spectrometry for the Characterisation and Bioanalysis of ADC...Quality Assistance s.a.
Dr. Arnaud Delobel from Quality Assistance spoke on advances in Mass Spectrometry for the Characterisation & Bioanalysis of ADCs at World ADC 2017 in Berlin.
For more on this topic, replay the full ADCs webinar here: http://bit.ly/2kW3KoN
With a full analytical package for Biologics, Quality Assistance provides scientific and technical support to biopharmaceutical companies developing Antibody-Drug Conjugates.
Specificaly, we can develop and/or optimise the analytical methods for a product and, if necessary, validate them according to international guidelines for batch release, stability testing and/or regulated bioanalysis.
For more information on our expertise and services, visit: www.quality-assistance.com
Follow us on social media:
LinkedIn: https://www.linkedin.com/company/quality-assistance
Twitter: https://twitter.com/QA_Belgium
Facebook: https://www.facebook.com/QualityAssistanceBelgium
Google +: https://plus.google.com/103676189647965359292
Quality Assistance S.A. is a leading European Contract Research Organisation providing the pharmaceutical industry with all the analytical services required by EMA and FDA regulations for the development and marketing of innovative human medicinal products.
We assist our clients from candidate selection, through non-clinical and clinical studies, to marketing authorisation, using our state-of-the-art, product-dedicated expertise in analytical sciences.
For each customer and each project, we design customised solutions, define analytical protocols, develop and validate specific new analytical methods and perform characterisation, stability, pharmacokinetic, biomarker and immunogenicity studies as well as batch release testing, in order to evaluate the Quality, Safety and Efficacy of the given drugs.
QIAseq Targeted DNA, RNA and Fusion Gene PanelsQIAGEN
Tumor heterogeneity has been known for a while but quantifying heterogeneity is still a challenge. NGS is the method of choice in the analysis of tumor heterogeneity, however, there are some inherent challenges associated with it. These include false positives, gaps in the gene due to overrepresentation and incomplete representation of low-frequency transcripts – all contributing to an inaccurate picture. Conventional library prep strategies for NGS are based on PCR, which introduces sequence-based bias and amplification noise, leading to these inaccuracies. In this webinar, we will cover
1. Principles of UMI and the new QIAseq product porfolio
2. How UMI along with SPE (single primer extension) allows for increased uniformity across difficult-to-sequence regions, removal of library construction bias, improved data analysis and sequencing optimization
3. How data generated from using UMI and SPE is directly comparable to analysis derived from whole transcriptome and exome sequencing
4. Application of UMI and SPE in the discovery of novel gene fusions and in the analysis of gene expression and genetic variation
Target enrichment enables researchers to focus their next generation sequencing (NGS) efforts on regions of interest, allowing them to obtain more sequencing data relevant to their study. In-solution target capture is a method of enrichment using oligonucleotide probes directed to specific regions within a genome. Target capture can be used to enrich multiple samples simultaneously, reducing the cost per sample, while using individually synthesized probes allows researchers to construct gene panels that can be optimized over time.
On The Detection of Ctni - A Comparison of Surface-Plasmon Optical - Electroc...komalicarol
“Cardiovascular diseases (CVDs) are the number one cause of
death globally, taking an estimated 17.9 million lives each year.
CVDs are a group of disorders of the heart and blood vessels and
include coronary heart disease, cerebrovascular disease, rheumatic
heart disease, and other conditions
A large amount of data is available on the functional impact of missense mutations in TP53 tumor suppressor gene. and on mutation patterns in many different cancers. TP53 direct sequencing is a time-consuming method with limitations in detection level. Here we describe the development and verification of a TP53 Next Generation Sequencing method based on Ion AmpliSeq technology. Preliminary data demonstrate that the TP53 Ion AmpliSeq panel and Ion Reporter Software analysis solution meets the requirements of clinical research laboratories.
“N2MO provides cutting-edge ex vivo insect based screening models for predicting ADMET properties of chemical substances in the early drug discovery phase. Insect models represents excellent model systems due to biological active test situation, combined with fast reproducibility with no need for resynthesizing compound and at same time the model produces data that are reflecting mammalian ADMET properties”
Flow Cytometry Training talks - part 1
This forms the first session of the Garvan Flow , Flow Cytometry Training course. this is a 1 1/2 day training course aimed at giving new and experienced researchers a better understanding of cytometry in medical and biological research.
Next generation sequencing (NGS) of circulating tumor DNA (ctDNA) from patient plasma is becoming more widespread in oncology clinical trials. The noninvasive nature of acquiring these samples is particularly important when resection of representative tumor samples is not advised or not possible. However, profiling of ctDNA has challenges to overcome, such as low concentration of ctDNA shed from the tumor and a low signal:noise ratio caused by somatic alterations with less than 1% variant allele fraction. Improving the sensitivity of these assays to detect low allele frequency events with high confidence requires robust sequencing of low input libraries while employing error correction to reduce background noise. To overcome these challenges, we have incorporated unique molecular identifiers (UMIs) into our NGS workflow. Using these novel adapters paired with our proprietary bioinformatics pipeline (AstraZeneca), the number of false positive variants reported for allele fractions less than 0.5% was reduced tenfold. We also refined our curation based on the mapping quality and strand bias in the vicinity of each variant to further reduce the background noise. The use of xGen® Dual Index UMI Adapters—Tech Access (Integrated DNA Technologies) has enabled us to sequence thousands of plasma samples from diverse tumor indications and at differing time points during our trials. The generated data are highly informative with the potential to answer critical questions relating to individual response or resistance to experimental therapies. During this webinar, we discuss our current NGS ctDNA workflow and our future plans to increase our sequencing sensitivity with these novel UMI adapters.
Advances in Mass Spectrometry for the Characterisation and Bioanalysis of ADC...Quality Assistance s.a.
Dr. Arnaud Delobel from Quality Assistance spoke on advances in Mass Spectrometry for the Characterisation & Bioanalysis of ADCs at World ADC 2017 in Berlin.
For more on this topic, replay the full ADCs webinar here: http://bit.ly/2kW3KoN
With a full analytical package for Biologics, Quality Assistance provides scientific and technical support to biopharmaceutical companies developing Antibody-Drug Conjugates.
Specificaly, we can develop and/or optimise the analytical methods for a product and, if necessary, validate them according to international guidelines for batch release, stability testing and/or regulated bioanalysis.
For more information on our expertise and services, visit: www.quality-assistance.com
Follow us on social media:
LinkedIn: https://www.linkedin.com/company/quality-assistance
Twitter: https://twitter.com/QA_Belgium
Facebook: https://www.facebook.com/QualityAssistanceBelgium
Google +: https://plus.google.com/103676189647965359292
Quality Assistance S.A. is a leading European Contract Research Organisation providing the pharmaceutical industry with all the analytical services required by EMA and FDA regulations for the development and marketing of innovative human medicinal products.
We assist our clients from candidate selection, through non-clinical and clinical studies, to marketing authorisation, using our state-of-the-art, product-dedicated expertise in analytical sciences.
For each customer and each project, we design customised solutions, define analytical protocols, develop and validate specific new analytical methods and perform characterisation, stability, pharmacokinetic, biomarker and immunogenicity studies as well as batch release testing, in order to evaluate the Quality, Safety and Efficacy of the given drugs.
QIAseq Targeted DNA, RNA and Fusion Gene PanelsQIAGEN
Tumor heterogeneity has been known for a while but quantifying heterogeneity is still a challenge. NGS is the method of choice in the analysis of tumor heterogeneity, however, there are some inherent challenges associated with it. These include false positives, gaps in the gene due to overrepresentation and incomplete representation of low-frequency transcripts – all contributing to an inaccurate picture. Conventional library prep strategies for NGS are based on PCR, which introduces sequence-based bias and amplification noise, leading to these inaccuracies. In this webinar, we will cover
1. Principles of UMI and the new QIAseq product porfolio
2. How UMI along with SPE (single primer extension) allows for increased uniformity across difficult-to-sequence regions, removal of library construction bias, improved data analysis and sequencing optimization
3. How data generated from using UMI and SPE is directly comparable to analysis derived from whole transcriptome and exome sequencing
4. Application of UMI and SPE in the discovery of novel gene fusions and in the analysis of gene expression and genetic variation
Target enrichment enables researchers to focus their next generation sequencing (NGS) efforts on regions of interest, allowing them to obtain more sequencing data relevant to their study. In-solution target capture is a method of enrichment using oligonucleotide probes directed to specific regions within a genome. Target capture can be used to enrich multiple samples simultaneously, reducing the cost per sample, while using individually synthesized probes allows researchers to construct gene panels that can be optimized over time.
On The Detection of Ctni - A Comparison of Surface-Plasmon Optical - Electroc...komalicarol
“Cardiovascular diseases (CVDs) are the number one cause of
death globally, taking an estimated 17.9 million lives each year.
CVDs are a group of disorders of the heart and blood vessels and
include coronary heart disease, cerebrovascular disease, rheumatic
heart disease, and other conditions
A large amount of data is available on the functional impact of missense mutations in TP53 tumor suppressor gene. and on mutation patterns in many different cancers. TP53 direct sequencing is a time-consuming method with limitations in detection level. Here we describe the development and verification of a TP53 Next Generation Sequencing method based on Ion AmpliSeq technology. Preliminary data demonstrate that the TP53 Ion AmpliSeq panel and Ion Reporter Software analysis solution meets the requirements of clinical research laboratories.
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
HOT NEW PRODUCT! BIG SALES FAST SHIPPING NOW FROM CHINA!! EU KU DB BK substit...GL Anaacs
Contact us if you are interested:
Email / Skype : kefaya1771@gmail.com
Threema: PXHY5PDH
New BATCH Ku !!! MUCH IN DEMAND FAST SALE EVERY BATCH HAPPY GOOD EFFECT BIG BATCH !
Contact me on Threema or skype to start big business!!
Hot-sale products:
NEW HOT EUTYLONE WHITE CRYSTAL!!
5cl-adba precursor (semi finished )
5cl-adba raw materials
ADBB precursor (semi finished )
ADBB raw materials
APVP powder
5fadb/4f-adb
Jwh018 / Jwh210
Eutylone crystal
Protonitazene (hydrochloride) CAS: 119276-01-6
Flubrotizolam CAS: 57801-95-3
Metonitazene CAS: 14680-51-4
Payment terms: Western Union,MoneyGram,Bitcoin or USDT.
Deliver Time: Usually 7-15days
Shipping method: FedEx, TNT, DHL,UPS etc.Our deliveries are 100% safe, fast, reliable and discreet.
Samples will be sent for your evaluation!If you are interested in, please contact me, let's talk details.
We specializes in exporting high quality Research chemical, medical intermediate, Pharmaceutical chemicals and so on. Products are exported to USA, Canada, France, Korea, Japan,Russia, Southeast Asia and other countries.
New Drug Discovery and Development .....NEHA GUPTA
The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
263778731218 Abortion Clinic /Pills In Harare ,sisternakatoto
263778731218 Abortion Clinic /Pills In Harare ,ABORTION WOMEN’S CLINIC +27730423979 IN women clinic we believe that every woman should be able to make choices in her pregnancy. Our job is to provide compassionate care, safety,affordable and confidential services. That’s why we have won the trust from all generations of women all over the world. we use non surgical method(Abortion pills) to terminate…Dr.LISA +27730423979women Clinic is committed to providing the highest quality of obstetrical and gynecological care to women of all ages. Our dedicated staff aim to treat each patient and her health concerns with compassion and respect.Our dedicated group ABORTION WOMEN’S CLINIC +27730423979 IN women clinic we believe that every woman should be able to make choices in her pregnancy. Our job is to provide compassionate care, safety,affordable and confidential services. That’s why we have won the trust from all generations of women all over the world. we use non surgical method(Abortion pills) to terminate…Dr.LISA +27730423979women Clinic is committed to providing the highest quality of obstetrical and gynecological care to women of all ages. Our dedicated staff aim to treat each patient and her health concerns with compassion and respect.Our dedicated group of receptionists, nurses, and physicians have worked together as a teamof receptionists, nurses, and physicians have worked together as a team wwww.lisywomensclinic.co.za/
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
1. Round Table Discussion
Thoughts on APC assay optimization -
Strengths and weaknesses of APC instruments
Prof. Clemens Möller, PhD
Albstadt-Sigmaringen University of Applied Sciences
office@biophysicalconsulting.de
www.clemensmoller.de
Nanion Usergroup Meeting
Sept 29, 2011
3. Controlled
state of
channel
Low binding
of
hydrophobic
compounds
Low leak
currents
Control of
membrane
potential /
capacitance
Patch-
Clampers
desire to
tinker with
the
experiment
PatchLiner, Port-a-Patch & SyncroPatch
data are in excellent correlation to MPC
Continuous
voltage
control
Chips made of
glass
substrate
Gigaseals
RS
compensation
HEKA
Software for
experiment
and data
evaluation
3
Experiments are performed under similar conditions as in MPC
4. Controlled
state of
channel
Low binding
of
hydrophobic
compounds
Low leak
currents
Control of
membrane
potential /
capacitance
Patch-
Clampers
desire to
tinker with
the
experiment
PatchLiner, Port-a-Patch & SyncroPatch
data are in excellent correlation to MPC
Continuous
voltage
control
Chips made of
glass
substrate
Gigaseals
RS
compensation
HEKA
Software for
experiment
and data
evaluation
4
Experiments are performed under similar conditions as in MPC
Main difference to MPC: Cells are delivered from suspension (not adherent).
Pharmacology? Networks of cells?
6. 6
Excellent correlation between manual and planar patch clamp
Before employing automated (planar)
patch-clamping in our programs, the
devices were validated with reference
and actual program compounds.
57 compounds within one chemical
series were tested side by side on
manual rigs and a planar patch clamp
device (PatchLiner)
Only 5 out of 57 compounds showed a
difference in the IC50-values of ~5-fold
Correlation: Manual Patch-Clamp vs PatchLiner
Manual Patch-Clamp IC50 [µM]
0.1 1 10 100
PatchLinerIC50[µM]
0.1
1
10
100
5
52
Pharmacological comparison of
Electrophysiology Platforms
PatchLiner validation
Reference: Davenport et al., 2010
7. Key points for pharmacology: same as for MPC and HT systems
o Prepare compound solutions as freshly as possible. Observe solubility of
compounds.
o Compound stock solutions required? How long are the compounds stable in
DMSO, at which storage temperature?
o Store solutions with reduced vehicle (DMSO) content in glassware, for as
short period as possible.
o Are currents stable under negative control conditions? Any vehicle effects?
o Do currents reach steady state in presence of compounds? (No continuous
perfusion in APC; repeated cpd administration required?)
o Consider pulse protocols (do the compounds exhibit preference for certain
states of the channel?)
o Consider temperature effects
7
PatchLiner, Port-a-Patch & SyncroPatch
data are in excellent correlation to MPC
8. (Many) Port-A-Patch, PatchLiner and
SyncroPatch assays are easy to set up
o Operation in standard modes easy
o Very "forgiving" cell culture
o Good seal success rates can be achieved with
suboptimal cells
o But: Seal enhancer (for most cell types)
appears to be required for good success
rates?
8
Eccellent seal success rates
Kv1.5
HERG
1st tier profilingExample: Panel of cardiac channels on PL
Kv1.1
Kv4.3/KChIP2
NaV1.5
L-type Ca2+
Standard protocols for most cell types
and channels are available.
For many cells, excellent success rates
can be achieved.
Current traces from Möller et al., 2010
9. (Many) Port-A-Patch, PatchLiner and
SyncroPatch assays are easy to set up
o The healthier the cells are, the better the seal success rate will typically be
o Cell confluency ~60-80%. Can depend on cell type
o Especially small / large cells? Consider different chip hole size
o Cell density appears to be not so critical (1 x 106 – 5 x 107 cells/ml are good
standard densities, but much lower densities have worked fine for some cells)
o Relatively small effect of pressure etc. settings in PatchControl software; standard
settings are often a good choice
9
Key points to consider for a good seal success rate
10. APC instruments complement each other
In addition, the Port-A-Patch proved very useful for
o Assay development support for PatchLiner and SyncroPatch
o Verification of data for compounds that showed inconsistent IC50 values on
the PatchLiner or the SyncroPatch
10
Instruments for different needs of throughput
Port-a-Patch PatchLiner SyncroPatch
11. APC instruments are highly flexible
11
Recordings from primary cells possible
300 nM Haloperidol
Negative control
Neurons – MAP2
astrocytes – GFAP
Nuclei – DRAQ5
Neurons Cardiomyocytes
Na+
Ca2+
K+
Reference: Möller et al., 2010
12. Modes of operation
12
Excised patch recordings not (yet, really) possible by planar APC
"Whole-
cell"
• Most widely used for pharmacology
"Cell-
attached
"
• Possible with some cells
"excised
patch"
• Are you missing single channel recordings
by APC? For MoA?
13. o Different features available
o Voltage clamp, current clamp (action potential recordings)
o Whole cell, perforated patch (are you using this a lot?)
o Intracellular solution exchange
o Fast ligand exchange (~50 ms)
o Temperature control
o Interaction during experiment possible
o Patch-Clampers desire to "play around" with an experiment
o Also, a "screening mode" with limited user access to settings is possible
(Talk by Corinna from last years meeting)
Different features available
APC instruments are highly flexible
13
Many features available; interaction possible
Interaction during experiment possible
14. Thank you for your attention and input!
Andreas Ebneth
Rainer Netzer
Heike Deisemann
Desireé Amm
York Rudhard
John Kemp
The whole great team,
especially:
Niels, Andrea, Michael,
Claudia, Sonja, Timo, Ali &
Cecilia
Ralf Kettenhofen
Martin Stolz
Prof. Clemens Möller, PhD | Albstadt-Sigmaringen University of Applied Sciences
clemens@biophysicalconsulting.de | www.clemensmoller.de
14
Carsten Claussen
Clemens Möller | Albstadt-Sigmaringen University of Applied Sciences
office@biophysicalconsulting.de | www.clemensmoller.de
15. References & Recommended Reading
15
References:
- Clemens Möller (2010). Keeping the Rhythm: hERG and beyond in Cardiovascular Safety Pharmacology. Expert Reviews Clinical
Pharmacology (Ion Channels) 3: 3. 321-329 May
- Adam J Davenport, Clemens Möller, Alexander Heifetz, Michael P Mazanetz, Richard J Law, Andreas Ebneth, Mark J Gemkow
(2010). Using Electrophysiology and in silico 3D Modelling to reduce hERG inhibition in a Histamine H3 Receptor Antagonist Program.
ASSAY and Drug Development Technologies 8: 6. 781-789 December
- Clemens Möller, Mark Slack (2010). Impact of new technologies for cellular screening along the drug value chain. Drug Discovery
Today 15: 9-10. 384-390 May
- Clemens Möller, Harry Witchel. Automated Electrophysiology Makes the Pace for Cardiac Ion Channel Safety Screening. (Submitted to
Frontiers, 2011)
Recommended reading:
Carol J Milligan, Li J, Sukumar P, Majeed Y, Dallas ML, English A, Emery P, Porter KE, Smith AM, McFadzean I, Beccano-Kelly
D, Bahnasi Y, Cheong A, Naylor J, Zeng F, Liu X, Gamper N, Jiang LH, Pearson HA, Peers C, Robertson B, Beech DJ (2009). Robotic
multiwell planar patch-clamp for native and primary mammalian cells. Nat Protoc. 2009;4(2):244-55.