Search results for: psychotropic drugs

Authors: Mohsen Shati, Seyede Salehe Mortazavi, Seyed Kazem Malakouti, Hamidreza Khanke Fazlollah Ahmadi

Abstract:

Taking unnecessary or excessive medication or using drugs with no indication (polypharmacy) by people of all ages, especially the elderly, is associated with increased adverse drug reactions (ADR), medical errors, hospitalization and escalating the costs. It may be facilitated or impeded by the healthcare system. In this study, we are going to describe the role of the health system in the practice of polypharmacy in Iranian elderly. In this Inductive qualitative content analysis using Graneheim and Lundman methods, purposeful sample selection until saturation has been made. Participants have been selected from doctors, pharmacists, policy-makers and the elderly. A total of 25 persons (9 men and 16 women) have participated in this study. Data analysis after incorporating codes with similar characteristics revealed 14 subcategories and six main categories of the referral system, physicians’ accessibility, health data management, drug market, laws enforcement, and social protection. Some of the conditions of the healthcare system have given rise to polypharmacy in the elderly. In the absence of a comprehensive specialty and subspecialty referral system, patients may go to any physician office so may well be confused about numerous doctors' prescriptions. Electronic records not being prepared for the patients, failure to comply with laws, lack of robust enforcement for the existing laws and close surveillance are among the contributing factors. Inadequate insurance and supportive services are also evident. Age-specific care providing has not yet been institutionalized, while, inadequate specialist workforce playing a major role. So, one may not ignore the health system as contributing factor in designing effective interventions to fix the problem.

Keywords: elderly, polypharmacy, health system, qualitative study

Procedia PDF Downloads 149

Authors: Khushbu Priyadarshi, Chandramani Pathak

Abstract:

Cancer cells can develop resistance to conventional therapies especially chemotherapeutic drugs. Resistance to chemotherapy is another challenge in cancer therapeutics. Therefore, it is important to address this issue. Gallic acid (GA) is a natural plant compound that exhibits various biological properties including anti-proliferative, anti-inflammatory, anti-oxidant and anti-bacterial. Despite of the wide spectrum biological properties GA has cytotoxic response and low bioavailability. To overcome this problem, GA was conjugated with the Polyamidoamine(PAMAM) dendrimer for improving the bioavailability and efficient delivery in drug-resistant HCT-116 Colon Cancer cells. Gallic acid was covalently linked to 4.0 G PAMAM dendrimer. PAMAM dendrimer is well established nanocarrier but has cytotoxicity due to presence of amphiphilic nature of amino group. In our study we have modified surface of PAMAM dendrimer with Gallic acid and examine their anti-proliferative effects in drug-resistant HCT-116 cells. Further, drug-resistant colon cancer cells were established and thereafter treated with different concentration of PAMAM-GA to examine their anti-proliferative potential. Our results show that PAMAM-GA conjugate induces apoptotic cell death in HCT-116 and drug-resistant cells observed by Annexin-PI staining. In addition, it also shows that multidrug-resistant drug transporter P-gp protein expression was downregulated with increasing the concentration of GA conjugate. After that we also observed the significant difference in Rh123 efflux and accumulation in drug sensitive and drug-resistant cancer cells. Thus, our study suggests that conjugation of anti-cancer agents with PAMAM could improve drug resistant property and cytotoxic response to treatment of cancer.

Keywords: drug resistance, gallic acid, PAMAM dendrimer, P-glycoprotein

Procedia PDF Downloads 148

Authors: Xiaolin Li, Terence Turney, John Forsythe, Bryce Feltis, Paul Wright, Vinh Truong, Will Gates

Abstract:

Clay-hydrogel nanocomposites have attracted great attention recently, mainly because of their enhanced mechanical properties and ease of fabrication. Moreover, the unique platelet structure of clay nanoparticles enables the incorporation of bioactive molecules, such as proteins or drugs, through ion exchange, adsorption or intercalation. This study seeks to improve the mechanical and rheological properties of a novel hydrogel system, copolymerized from a tetrapodal polyethylene glycol (PEG) thiol and a linear, triblock PEG-PPG-PEG (PPG: polypropylene glycol) α,ω-bispropynoate polymer, with the simultaneous incorporation of various amounts of Na-saturated, montmorillonite clay (MMT) platelets (av. lateral dimension = 200 nm), to form a bioactive three-dimensional network. Although the parent hydrogel has controlled swelling ability and its PEG groups have good affinity for the clay platelets, it suffers from poor mechanical stability and is currently unsuitable for potential applications. Nanocomposite hydrogels containing 4wt% MMT showed a twelve-fold enhancement in compressive strength, reaching 0.75MPa, and also a three-fold acceleration in gelation time, when compared with the parent hydrogel. Interestingly, clay nanoplatelet incorporation into the hydrogel slowed down the rate of its dehydration in air. Preliminary results showed that protein binding by the MMT varied with the nature of the protein, as horseradish peroxidase (HRP) was more strongly bound than bovine serum albumin. The HRP was no longer active when bound, presumably as a result of extensive structural refolding. Further work is being undertaken to assess protein binding behaviour within the nanocomposite hydrogel for potential diabetic wound healing applications.

Keywords: hydrogel, nanocomposite, small molecule, wound healing

Procedia PDF Downloads 267

Authors: Huma Shareef, Ghazala H. Rizwani, Ahsana Dar

Abstract:

In traditional medicine Cardiospermum halicacabum L. (Sapindeaceae) is used against various ailments. In current investigation searching a new remedy that will available easily, non expensive, able to lower hypertension and standardize blood pressure, made us to develop an herbal medicine. Crude ethanol extract of C. halicacabum and its various fractions ethyl acetate and butanol showed a dose-dependent hypotensive effect in anaesthetized rats. The trachea was exposed and freed from connective tissue and incubated by cannula to facilitate spontaneous respiration. The right carotid artery and left jugular vein were cannulated with polyethylene tubing PE-50 for monitoring blood pressure changes via pressure transducer (Gould P23 ID) connected to a Grass model 79D polygraph and for i.v. injection, respectively. Drugs or the plant extracts were administered at a constant volume of 0.5 ml/kg, followed by injection of 0.2 ml of saline that flushed the cannula. Systolic, diastolic and mean arterial blood pressure (MABP) was measured in mm Hg and heart rate in beats/min. Ethanol extract of C. halicacabum showed a significant activity at 50 mg/kg dose. Ethyl acetate fraction (10, 20, 30, 40, and 50 mg/kg) induced dose dependent fall in systolic and diastolic blood pressure, heart rate of rats. At 10-30 mg/kg the hypotensive effect was non significantly reduced by 10 -15%. However, the extract at 40 mg/kg induced significant hypotensive effect calculated as 30.95±3.2% MABP and this effect persists till 50 mg/kg. The higher polar fraction (butanol) of the whole plant failed to produce any significant response against MABP at all the tested doses (10-50 mg/kg). C. halicacabum lowers blood pressure, exerts a dose-dependent hypotensive effect, can be used as hypotensor.

Keywords: cardiospermum halicacabum, calcium channel blocker, hypotensive, various extracts

Procedia PDF Downloads 502

Authors: Halilu Babaji, Adamu Buba

Abstract:

The proliferation of small arms and light weapons (SALW) and its illicit trafficking in West Africa and Nigeria in particular, pose a major threat to peace, security and development in the Sub-region. The high circulation of these weapons in the region is a product of the interplay of several factors, which derives principally from the internal socio-economic and political dynamics compounded by globalization. The process of globalization has congealed both time and space making it easier for ideas, goods, persons, services, information, products and money to move across borders with fewer restrictions. And this has a negative effect in the entire region making it easier for arms, ammunition, insurgents, criminal and drugs to flow within national boundaries. The failure of public security in most parts of Nigeria has lead communities to indulge in different forms of ‘self-help ‘security measures, ranging from vigilante groups to community-owned arms stockpiling. Having lost confidence in the Nigerian state, parties to some of these conflicts have become entangled in a security dilemma. The quest to procure more arms to guarantee personal and community protection from perceived and real enemies is fuelling the ‘domestic arms race ‘. Therefore, as small arms remain-and proliferate – development is impeded. The impact of SALW on economic well being and national development in Nigeria is of vast significant. Therefore the need to collect these arms in circulation in Nigeria particularly the volatile area of North-east is of very important. This will hopefully contribute to government effort in building a free, secured and peaceful society.

Keywords: arms, development, proliferation, security

Procedia PDF Downloads 325

Authors: Mani Mofidi, Neda Valizadeh, Ali Hashemaghaee, Mona Hashemaghaee, Soudabeh Shafiee Ardestani

Abstract:

Background: Acute pain and its management remained the most complaint of emergency service admission. Diagnostic and therapeutic procedures add to patients’ pain. Diminishing the pain increases the quality of patient’s feeling and improves the patient-physician relationship. Aim: The aim of this study was to evaluate the outcomes and side effects of opioid administration in emergency patients. Material and Methods: patients admitted to ward II emergency service of Imam Khomeini hospital, who received one of the opioids: morphine, pethidine, methadone or fentanyl as an analgesic were evaluated. Their vital signs and general condition were examined before and after drug injection. Also, patient’s pain experience were recorded as numerical rating score (NRS) before and after analgesic administration. Results: 268 patients were studied. 34 patients were addicted to opioid drugs. Morphine had the highest rate of prescription (86.2%), followed by pethidine (8.5%), methadone (3.3%) and fentanyl (1.68). While initial NRS did not show significant difference between addicted patients and non-addicted ones, NRS decline and its score after drug injection were significantly lower in addicted patients. All patients had slight but statistically significant lower respiratory rate, heart rate, blood pressure and O2 saturation. There was no significant difference between different kind of opioid prescription and its outcomes or side effects. Conclusion: Pain management should be always in physicians’ mind during emergency admissions. It should not be assumed that an addicted patient complaining of pain is malingering to receive drug. Titration of drug and close monitoring must be in the curriculum to prevent any hazardous side effects.

Keywords: numerical rating score, opioid, pain, emergency department

Procedia PDF Downloads 426

Authors: Azza H. El-Medany, Hanan H. Hagar, Omnia A. Nayel, Jamila H. El-Medany

Abstract:

In search for drugs that can target cancer cell micro-environment in as much as being able to halt malignant cellular transformation, the natural dietary phytochemical curcumin was currently assessed in DMH-induced colorectal cancer rat model. The study enrolled 50 animals divided into a control group (n=10) and DMH-induced colorectal cancer control group (n=20) (20mg/kg-body weight for 28 weeks) versus curcumin-treated group (n=20) (160 mg/kg suspension daily oral for further 8 weeks). Treatment by curcumin succeeded to significantly decrease the percent of ACF and tended to normalize back the histological changes retrieved in adenomatous and stromal cells induced by DMH. The drug also significantly elevated GSH and significantly reduced most of the accompanying biochemical elevations (namely MDA, TNF-α, TGF-β and COX2) observed in colonic carcinomatous tissue, induced by DMH, thus succeeding to revert that of MDA, COX2 and TGF-β back to near normal as justified by being non-significantly altered as compared to normal controls. The only exception was PAF that was insignificantly altered by the drug. When taken together, it could be concluded that curcumin possess the potentiality to halt some of the orchestrated cross-talk between cancerous transformation and its micro-environmental niche that contributes to cancer initiation, progression and metastasis in this experimental cancer colon model. Envisioning these merits to a drug with already known safety preferentiality, awaits final results of current ongoing clinical trials, before curcumin can be added to the new therapeutic armamentarium of anticancer therapy.

Keywords: curcumin, dimethyl hydralazine, aberrant crypt foci, malondialdehyde, reduced glutathione, cyclooxygenase-2, tumour necrosis factor-alpha, transforming growth factor-beta, platelet activating factor

Procedia PDF Downloads 294

Authors: Kamel Abd Elaziz Mohamed

Abstract:

Introduction: Drug related problems (DRPs) are of major concern, affecting patients of both sex. They impose considerable economic burden on the society and the health-care systems. Aim of the work: The aim of this work was to identify and categorize drug-related problems in adult intensive care unit. Patients and methods: The study was a prospective, observational study as eighty six patients were included. They were consecutively admitted to ICU through the emergency room or transferred from the general ward due to DRPs. Parameters included in the study as length of stay in ICU, need for cardiovascular support or mechanical ventilation, dialysis, as well as APACHE II score were recorded. Results: Drug related problems represent 3.6% of the total ICU admission. The median (range) of APACHE II score for 86 patients included in the study was 17 (10-23), and length of ICU stay was 2.4 (1.5-4.2) days. In 45 patients (52%), DRP was drug over dose (group 1), while other DRP was present in the other 41 patients (48%, group 11). Patients in group 1 were older (39 years versus 32 years in group 11), with significant impaired renal function. The need of inotropic drugs and mechanical ventilation as well as the length of stay (LOS) in ICU was significantly higher in group 1. There were no significant difference in GCS between both groups, however APACHE II score was significantly higher in group 1. Only four patients (4.6%) were admitted by suicidal attempt as well as three patients (3.4%) due to trauma drug-related admissions, all were in (group 1). Nineteen percent of the patients had drug related problem due to hypoglycaemic medication followed by tranquilizer (15%). Adverse drug effect followed by failure to receive medication were the most causes of drug problem in (group11).The total mortality rate was 4.6%, all of them were eventually non preventable. Conclusion: The critically ill patients admitted due to drug related problems represented a small proportion (3.6%) of admissions to the ICU. Hypoglycaemic medication was one of the most common causes of admission by drug related problems.

Keywords: drug related problems, ICU, cost, safety

Procedia PDF Downloads 332

Authors: D. Farsi, G. Dokhtvasi, S. Abbasi, S. Shafiee Ardestani, E. Payani

Abstract:

Background and aim:It has not been well studied whether fentanyl-thiopental (FT) is effective and safe for PSA in orthopedic procedures in Emergency Department (ED). The aim of this trial was to evaluate the effectiveness of intravenous FTversusfentanyl-midazolam (FM)in patients who suffered from shoulder dislocation or distal radial fracture-dislocation. Methods:In this randomized double-blinded study, Seventy-six eligible patients were entered the study and randomly received intravenous FT or FM. The success rate, onset of action and recovery time, pain score, physicians’ satisfaction and adverse events were assessed and recorded by treating emergency physicians. The statistical analysis was intention to treat. Results: The success rate after administrating loading dose in FT group was significantly higher than FM group (71.7% vs. 48.9%, p=0.04); however, the ultimate unsuccess rate after 3 doses of drugs in the FT group was higher than the FM group (3 to 1) but it did not reach to significant level (p=0.61). Despite near equal onset of action time in two study group (P=0.464), the recovery period in patients receiving FT was markedly shorter than FM group (P<0.001). The occurrence of adverse effects was low in both groups (p=0.31). Conclusion: PSA using FT is effective and appears to be safe for orthopedic procedures in the ED. Therefore, regarding the prompt onset of action, short recovery period of thiopental, it seems that this combination can be considered more for performing PSA in orthopedic procedures in ED.

Keywords: procedural sedation and analgesia, thiopental, fentanyl, midazolam, orthopedic procedure, emergency department, pain

Procedia PDF Downloads 251

Authors: Sajjad Jafari

Abstract:

Background and Aim: The use of herbal medicines has a long history. Today, due to the resistance of microorganisms to antibiotics and antimicrobial substances, herbal medicines have attracted attention due to their significant antimicrobial effects and low toxicity. This study aims to systematically review the antimicrobial effects of different plant extracts (Quercus infectoria) as possible candidates for treating infectious diseases. Material and Methods: The present study is a review study by searching reputable scientific databases such as PubMed, Google Scholar, Scopus, and Web of Science from 2000 to 2023 using the keywords Antimicrobial, Quercus infectoria, Medicinal herbal, Infectious diseases the latest information obtained. Results: In this study, 45 articles were found and reviewed. Quercus infectoria is a small tree native to Greece, Asia Minor, and Iran. Quercus is a plant genus in the family of Fagaceae. This species is generally known under the name ‘‘baloot” in Iran and is commonly used as a medicinal plant. The extracts used included water, hydro-alcoholic, ethanol, methanol. This plant had high inhibition activity and a lethal effect on gram-positive and gram-negative bacteria of ATCC strains, hospital, and resistant strains. Therefore, in addition to antibacterial effects, antiparasitic and antifungal effects. The seed of the plant was the most used and the most effective antimicrobial extract among the ethanol and methanol extracts. Conclusion: The findings of this study suggest that Quercus infectoria has significant antimicrobial effects against a wide range of microorganisms. This makes it a potential candidate for the development of new antimicrobial drugs. Further research is needed to confirm the efficacy and safety of Quercus infectoria in clinical trials.

Keywords: antimicrobial, Quercus infectoria, medicinal herbal, infectious diseases

Procedia PDF Downloads 94

Authors: Shivankar Agrawal, Sunil Kumar Deshmukh, Colin Barrow, Alok Adholeya

Abstract:

A variety of genetic and environmental factors cause various cosmetics and dermatological problems. There are already claimed drugs available in market for treating these problems. However, the challenge remains in finding more potent, environmental friendly, causing minimal side effects and economical cosmeceuticals. This leads to an increased demand for natural cosmeceutical products in the last few decades. Plant derived ingredients are limited because plants either contain toxic metabolites, grow too slow or seasonal harvesting is a problem. The research work carried out in this project aims at isolation, characterization of marine fungal secondary metabolite and evaluating their potential use in future cosmetic skin care products. We have isolated and purified 35 morphologically different fungal isolates from various marine habitats of the India. These isolates have been functionally characterized for anti-tyrosinase, antioxidant and anti-acne activities. For molecular characterization, the Internal Transcribed spacer (ITS) region of 15 functionally active marine fungal isolates was amplified using universal primers, ITS1 and ITS4 and sequenced. Out of 15 marine fungal isolates crude extract of strains D4 (Aspergillus terreus) and P2 (Talaromyces stipitatus) showed 70% and 57% tyrosinase inhibition at 1mg/mL respectively. Strain D5 (Simplicillium lamellicola) has showed significant inhibition against Propionibacterium acnes and Staphylococcus epidermidis. In addition, all these strains also displayed DPPH- radical scavenging activity and may be utilized as skin cosmeceutical applications. Purification and characterization of crude extracts for identification of active lead molecule is under process.

Keywords: anti-acne, anti-tyrosinase, cosmeceutical, marine fungi

Procedia PDF Downloads 275

Authors: Monika Kallubai, Shreya Dubey, Rajagopal Subramanyam

Abstract:

Our study is all about the biological interactions of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules with carrier protein Human Serum Albumin (HSA). The cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 µM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. The further experiment proved that Dhc and DhcA induced 35.6% and 37.7% early apoptotic cells and 2.5%, 2.9% late apoptotic cells respectively. Morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7±0.03×104 M-1 and 3.9±0.05×104 M-1, and their binding free energies were found to be -6.4 and -6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4±0.05×104 M-1 replaced Dhc, and phenylbutazone 1.5±0.05×104 M-1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular sizes of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported the greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for the further development of steroid derivatives with improved pharmacological significance as novel anti-cancer drugs.

Keywords: apoptosis, dihydrocortisol, fluorescence quenching, protein conformations

Procedia PDF Downloads 129

Authors: Ahmed M. Debela, Mayreli Ortiz , Ciara K. O´Sullivan

Abstract:

The completion of the human genome Project (HGP) has paved the way for mapping the diversity in the overall genome sequence which helps to understand the genetic causes of inherited diseases and susceptibility to drugs or environmental toxins. Arrayed primer extension (APEX) is a microarray based minisequencing strategy for screening disease causing mutations. It is derived from Sanger DNA sequencing and uses fluorescently dideoxynucleotides (ddNTPs) for termination of a growing DNA strand from a primer with its 3´- end designed immediately upstream of a site where single nucleotide polymorphism (SNP) occurs. The use of DNA polymerase offers a very high accuracy and specificity to APEX which in turn happens to be a method of choice for multiplex SNP detection. Coupling the high specificity of this method with the high sensitivity, low cost and compatibility for miniaturization of electrochemical techniques would offer an excellent platform for detection of mutation as well as sequencing of DNA templates. We are developing an electrochemical APEX for the analysis of SNPs found in the MYH7 gene for group of cardiomyopathy patients. ddNTPs were labeled with four different redox active compounds with four distinct potentials. Thiolated oligonucleotide probes were immobilised on gold and glassy carbon substrates which are followed by hybridisation with complementary target DNA just adjacent to the base to be extended by polymerase. Electrochemical interrogation was performed after the incorporation of the redox labelled dedioxynucleotide. The work involved the synthesis and characterisation of the redox labelled ddNTPs, optimisation and characterisation of surface functionalisation strategies and the nucleotide incorporation assays.

Keywords: array based primer extension, labelled ddNTPs, electrochemical, mutations

Procedia PDF Downloads 245

Authors: Sharjeel Abid, Tanveer Hussain, Ahsan Nazir, Abdul Zahir, Nabyl Khenoussi

Abstract:

Localized application of drug has various advantages and fewer side effects as compared with other methods. Burn patients suffer from swear pain and the major aspects that are considered for burn victims include pain and infection management. Nano-fibers (NFs) loaded with drug, applied on local wound area, can solve these problems. Therefore, this study dealt with the fabrication of drug loaded NFs for better pain management. Two layers of NFs were fabricated with different drugs. Contact layer was loaded with Gabapentin (a nerve painkiller) and the second layer with acetaminophen. The fabricated dressing was characterized using scanning electron microscope, Fourier Transform Infrared Spectroscopy, X-Ray Diffraction and UV-Vis Spectroscopy. The double layered based NFs dressing was designed to have both initial burst release followed by slow release to cope with pain for two days. The fabricated nanofibers showed diameter < 300 nm. The liquid absorption capacity of the NFs was also checked to deal with the exudate. The fabricated double layered dressing with dual drug loading and release showed promising results that could be used for dealing pain in burn victims. It was observed that by the addition of drug, the size of nanofibers was reduced, on the other hand, the crystallinity %age was increased, and liquid absorption decreased. The combination of fast nerve pain killer release followed by slow release of non-steroidal anti-inflammatory drug could be a good tool to reduce pain in a more secure manner with fewer side effects.

Keywords: pain management, burn wounds, nano-fibers, controlled drug release

Procedia PDF Downloads 251

Authors: Nita Nurlaila Kadarwaty

Abstract:

Purpose: This study presents a rare case of bilateral Acute Primary Angle Closure Glaucoma (PACG). Method: A case report of a 64-year-old woman with a good outcome Acute PACG in both eyes who underwent phacotrabeculectomy surgery. Result: A 64-year-old woman complained of acute pain in both eyes, accompanied by decreased vision, photophobia, and seeing halos for three weeks. There was no history of trauma, steroid or other systemic drugs used, or intraocular surgery before. Ophthalmologic examination revealed a right eye (RE) visual acuity of 0.1, left eye (LE) 0.2. RE intraocular pressure (IOP) was 12 mmhg and LE: 36.4 mmHg in medication of timolol maleat ED and acetazolamide oral. Both eyes' anterior segments revealed mixed injection, corneal edema, shallow anterior chamber, posterior synechiae, mid-dilatation pupil with negative pupillary reflection, and cloudy lens without intumescent. There was a glaucomatous optic and closed iridocorneal angle on the gonioscopy. Initial treatments included oral acetazolamide and potassium aspartate 250 mg three times a day, timolol maleate ED 0.5% twice a day, and prednisolone acetate ED 1% four times a day. This patient underwent trabeculectomy, phacoemulsification, and implantation of IOL in both eyes. One week after the surgeries, both eyes showed decreased IOP and good visual improvement. Conclusion: Bilateral simultaneous Acute PACG is generally severe and results in a poor outcome. It causes rapidly progressive visual loss and is often irreversible. Phacotrabeculectomy has more benefits compared to only phacoemulsification for the intervention regarding the reduced IOP post-surgical.

Keywords: acute primary angle closure glaucoma, intraocular pressure, phacotrabeculectomy, glaucoma

Procedia PDF Downloads 72

Authors: Mona Hassan Aburahma, Alaa Hamed Salama

Abstract:

Ufasomes “unsaturated fatty acids liposomes” are unique nano-sized self-assembled bilayered vesicles that can be easily created from the readily available unsaturated fatty acid. Ufasomes are formed due to weak associative interaction of the fully ionized and unionized fatty acids into bilayers structures. In the ufasomes constructs, the fatty acid molecules are oriented with their hydrocarbon tails directed toward the membrane interior and the carboxyl groups are in contact with water. Although ufasomes can be employed as a safe vesicular carrier for drugs, the extreme instability of their aqueous dispersions hinders their effective use in drug delivery field. Accordingly, in our study, lyophilized gels containing ufasomes were prepared using a simple assembling technique form the readily available oleic acid to overcome the colloidal instability of the ufasomes dispersions and convert them into accurate unit dosage forms. The influence of changing cholesterol percentage relative to oleic acid on the ufasomes vesicles were investigated using factorial design. The optimized oleic acid ufasomes comprised nanoscaled spherical vesicles. Scanning electron micrographs of the lyophilized gels revealed that the included ufasomes were intact, non-aggregating, and preserved their spherical morphology. Rheological characterization (viscosity and shear stress versus shear rate) of reconstituted ufasomal lyophilized gel ensured the ease of application. The capability of the ufasomes, included in the gel, to penetrate deep through the mucosa layers was illustrated using ex-vivo confocal laser imaging, thereby, highlighting the feasibility of stabilizing ufasomes using lyophilized gel platforms.

Keywords: ufasomes, lyophilized gel, confocal scanning microscopy, rheological characterization, oleic acid

Procedia PDF Downloads 405

Authors: Minh-Phuong Ngoc Bui, Abdennour Abbas

Abstract:

Microbial spoilage of food/drug has been a constant nuisance and an unavoidable problem throughout history that affects food/drug quality and safety in a variety of ways. A simple and rapid test of fungi and bacteria in food/drugs and environmental clinical samples is essential for proper management of contamination. A number of different techniques have been developed for detection and enumeration of foodborne microorganism including plate counting, enzyme-linked immunosorbent assay (ELISA), polymer chain reaction (PCR), nucleic acid sensor, electrical and microscopy methods. However, the significant drawbacks of these techniques are highly demand of operation skills and the time and cost involved. In this report, we introduce a rapid method for detection of bacteria and fungi in food/drug products using a specific interaction between a reducing agent (tris(2-carboxylethyl)phosphine (TCEP)) and the microbial surface proteins. The chemical reaction was transferred to a transduction system using gold nanoplates-enhanced chemiluminescence. We have optimized our nanoplates synthetic conditions, characterized the chemiluminescence parameters and optimized conditions for the microbial assay. The new detection method was applied for rapid detection of bacteria (E.coli sp. and Lactobacillus sp.) and fungi (Mucor sp.), with limit of detection as low as single digit cells per mL within 10 min using a portable luminometer. We expect our simple and rapid detection method to be a powerful alternative to the conventional plate counting and immunoassay methods for rapid screening of microorganisms in food/drug products.

Keywords: microorganism testing, gold nanoplates, chemiluminescence, reducing agents, luminol

Procedia PDF Downloads 297

Authors: Devadrita Dey Sarkar

Abstract:

Oxidosqualene cyclase is a membrane bound enzyme in which helps in the formation of steroid scaffold in higher organisms. In a highly selective cyclization reaction oxidosqualene cyclase forms LANOSTEROL with seven chiral centres starting from the linear substrate 2,3-oxidosqualene. In humans OSC in cholesterol biosynthesis it represents a target for the discovery of novel anticholesteraemic drugs that could complement the widely used statins. The enzyme oxidosqualene: lanosterol cyclase (OSC) represents a novel target for the treatment of hypercholesterolemia. OSC catalyzes the cyclization of the linear 2,3-monoepoxysqualene to lanosterol, the initial four-ringed sterol intermediate in the cholesterol biosynthetic pathway. OSC also catalyzes the formation of 24(S), 25-epoxycholesterol, a ligand activator of the liver X receptor. Inhibition of OSC reduces cholesterol biosynthesis and selectively enhances 24(S),25-epoxycholesterol synthesis. Through this dual mechanism, OSC inhibition decreases plasma levels of low-density lipoprotein (LDL)-cholesterol and prevents cholesterol deposition within macrophages. The recent crystallization of OSC identifies the mechanism of action for this complex enzyme, setting the stage for the design of OSC inhibitors with improved pharmacological properties for cholesterol lowering and treatment of atherosclerosis. While studying and designing the inhibitor of oxidosqulene cyclase, I worked on the pdb id of 1w6k which was the most worked on pdb id and I used several methods, techniques and softwares to identify and validate the top most molecules which could be acting as an inhibitor for oxidosqualene cyclase. Thus, by partial blockage of this enzyme, both an inhibition of lanosterol and subsequently cholesterol formation as well as a concomitant effect on HMG-CoA reductase can be achieved. Both effects complement each other and lead to an effective control of cholesterol biosynthesis. It is therefore concluded that 2,3-oxidosqualene cyclase plays a crucial role in the regulation of intracellular cholesterol homeostasis. 2,3-Oxidosqualene cyclase inhibitors offer an attractive approach for novel lipid-lowering agents.

Keywords: anticholesteraemic, crystallization, statins, homeostasis

Procedia PDF Downloads 349

Authors: Mohammed Jarrar, Shalini Behl, Nadia Shaheen, Abeer Fatima, Reem Nasab

Abstract:

Skin aging is a slow multifactorial process influenced by both internal as well as external factors. Ultra-violet radiations (UV), diet, smoking and personal habits are the most common environmental factors that affect skin aging. Fat contents and fibrous proteins as collagen and elastin are core internal structural components. The direct influence of UV on elastin integrity and health is crucial on aging of skin by time. The deposition of abnormal elastic material is a major marker in a photo-aged skin. Searching for compounds that may protect against cutaneous photo-damage is highly valued. Retinoids and Alpha Hydroxy Acids protective and or repairing effects of UV have been endorsed by some researchers. For consolidating a better understanding of anti and protective effects of such anti-aging agents, we evaluated the combinatory effects of various dosages of lactic acid and retinol on the dermal fibroblasts elastin levels exposed to UV. The UV exposed cells showed significant reduction in the elastin levels. A combination of drugs with a higher concentration of lactic acid (30-35 mM) and a lower concentration of retinol (10-15mg/mL) showed to work better in enhancing elastin concentration in UV exposed cells. We assume this enhancement could be the result of increased tropo-elastin gene expression stimulated by retinol and lactic acid probably repaired the UV irradiated damage by enhancing the amount and integrity of the elastin fibers.

Keywords: alpha hydroxy acid, elastin, retinol, ultraviolet radiations

Procedia PDF Downloads 341

Authors: Olga Shapovalova, Vladimir Vinogradov

Abstract:

Among many different sol – gel matrices only alumina can be successfully parenteral injected in the human body. And this is not surprising, because boehmite (aluminium oxyhydroxide) is the metal oxide approved by FDA and EMA for intravenous and intramuscular administrations, and also has been using for a longtime as adjuvant for producing of many modern vaccines. In our earlier study, it has been shown, that denaturation temperature of enzymes entrapped in sol-gel boehmite matrix increases for 30 – 60 °С with preserving of initial activity. It makes such matrices more attractive for long-term storage of non-stable drugs. In current work we present ultrasound-assisted sol-gel synthesis of nano-boehmite. This method provides bio-friendly, very stable, highly homogeneous alumina sol with using only water and aluminium isopropoxide as a precursor. Many parameters of the synthesis were studied in details: time of ultrasound treatment, US frequency, surface area, pore and nanoparticle size, zeta potential and others. Here we investigated the dependence of stability of colloidal sols and textural properties of the final composites as a function of the time of ultrasonic treatment. Chosen ultrasonic treatment time was between 30 and 180 minutes. Surface area, average pore diameter and total pore volume of the final composites were measured by surface and pore size analyzer Nova 1200 Quntachrome. It was shown that the matrices with ultrasonic treatment time equal to 90 minutes have the biggest surface area 431 ± 24 m2/g. On the other had such matrices have a smaller stability in comparison with the samples with ultrasonic treatment time equal to 120 minutes that have the surface area 390 ± 21 m2/g. It was shown that the stable sols could be formed only after 120 minutes of ultrasonic treatment, otherwise the white precipitate of boehmite is formed. We conclude that the optimal ultrasonic treatment time is 120 minutes.

Keywords: boehmite matrix, stabilisation, ultrasound-assisted sol-gel synthesis

Procedia PDF Downloads 265

Authors: Laila Khawaja

Abstract:

In recent years, an increasing emphasis has been placed on measuring program implementation, in part because of the great variability in how complex interventions are delivered in real-life settings. There is an increased awareness that while conducting process evaluations, one should aim to identify and understand the complexities of intervention if they are to be used for future intervention development or the strategies needed to implement the same intervention in a different setting. Complex interventions are public health interventions that are not drugs or surgical procedures but have many potential active aspects of intervention. In this paper, process evaluations are aligned with MRC guidelines to identify contextual factors related to outcomes to assess the quality of implementation. This paper briefly discusses the perceptions of research team on the implementation of the intervention of ‘Engaging Adolescents in Changing Behaviour’ (EACH-B), a school-based complex intervention study aiming to improve diet and physical activity among adolescents aged 12-13 years. Through qualitative interviews and focus groups with 10 staff members, we aimed to understand their experiences and reflections on implementing the EACH-B trial delivered in 49 Schools around Hampshire, England. Data were uploaded into NVivo, and analysis was conducted using thematic analysis. The investigation revealed two overarching themes: (a) how the communication patterns with teachers were impacted during the delivery of implementation and (b) what were the team’s strategies to keep logistics aligned with the research process that impacted the overall implementation of the trial. The paper informs adaptation strategies used by the research team to establish and maintain effective communication with the teachers as well as the thoughtfulness of the team’s logistic strategy for the successful delivery of the trial.

Keywords: complex interventions, process evaluation, adaptation strategies, randomised controlled trial

Procedia PDF Downloads 63

Authors: Teka Haile, Behailu Hawulte, Solomon Alemayehu

Abstract:

Background: HIV virological failure still remains a problem in HV/AIDS treatment and care. This study aimed to describe the prevalence and identify the factors associated with viral non-suppression among HIV-positive adult patients on antiretroviral therapy in Woliso Town, Oromia, Ethiopia. Methods: A retrospective cross-sectional study was conducted among 424 HIV-positive patient’s attending antiretroviral therapy (ART) in Woliso Town during the period from August 25, 2020 to August 30, 2020. Data collected from patient medical records were entered into Epi Info version 2.3.2.1 and exported to SPSS version 21.0 for analysis. Logistic regression analysis was done to identify factors associated with viral load non-suppression, and statistical significance of odds ratios were declared using 95% confidence interval and p-value < 0.05. Results: A total of 424 patients were included in this study. The mean age (± SD) of the study participants was 39.88 (± 9.995) years. The prevalence of HIV viral load non-suppression was 55 (13.0%) with 95% CI (9.9-16.5). Second-line ART treatment regimen (Adjusted Odds Ratio (AOR) = 8.98, 95% Confidence Interval (CI): 2.64, 30.58) and routine viral load testing (AOR = 0.01, 95% CI: 0.001, 0.02) were significantly associated with virological non-suppression. Conclusion: Virological non-suppression was high, which hinders the achievement of the third global 95 target. The second-line regimen and routine viral load testing were significantly associated with virological non-suppression. It suggests the need to assess the effectiveness of antiretroviral drugs for epidemic control. It also clearly shows the need to decentralize third-line ART treatment for those patients in need.

Keywords: virological non-suppression, HIV-positive, ART, Woliso town, Ethiopia

Procedia PDF Downloads 146

Authors: Anita K. Kovacs, Peter Hegyes, Zsolt Bozso, Gabor Toth

Abstract:

Fluorination has been used extensively by the pharmaceutical industry as a strategy to improve the pharmacokinetics of drugs due to its effectiveness in increasing the potency of antimicrobial peptides (AMPs). Multiple-fluorinated indole-ring-containing tryptophan derivatives have the potential of having better antimicrobial activity than the widely used mono-fluorinated indole-ring containing tryptophan derivatives, but they are not available commercially. Therefore, our goal is to synthesize multiple-fluorinated indole-ring containing tryptophan derivatives to incorporate them into AMPs to enhance their antimicrobial activity. During our work, we are trying several methods (classical organic synthesis, enzymic synthesis, and solid phase peptide synthesis) for the synthesis of the said compounds, with mixed results. With classical organic synthesis (four different routes), we did not get the desired results. The reaction of serin with substituted indole in the presence of acetic anhydride led to racemic tryptophane; with the reaction of protected serin with indole in the presence of nickel complex was unsuccessful; the reaction of serin containing protected dipeptide with disuccinimidyl carbonate we achieved a tryptophane containing dipeptide, its chiral purity is being examined; the reaction of alcohol with substituted indole in the presence of copper complex was successful, but it was only a test reaction, we could not reproduce the same result with serine. The undergoing tryptophan-synthase method has shown some potential, but our work has not been finished yet. The successful synthesis of the desired multiple-fluorinated indole-ring-containing tryptophan will be followed by solid phase peptide synthesis in order to incorporate it into AMPs to enhance their antimicrobial activity. The successful completion of these phases will mean the possibility of manufacturing new, effective AMPs.

Keywords: halogenation, fluorination, tryptophan, enhancement of antimicrobial activity

Procedia PDF Downloads 95

Authors: Anindita Sen

Abstract:

Oral fluid is a promising diagnostic matrix for drugs of abuse compared to urine and serum. Detection of methamphetamine in oral fluid would pave way for the easy evaluation of impairment in drivers during roadside drug testing as well as ensure safe working environments by facilitating evaluation of impairment in employees at workplaces. A membrane-based point-of-care (POC) friendly pre-treatment technique has been developed which aided elimination of interferences caused by salivary proteins and facilitated the demonstration of methamphetamine detection in saliva using a gold nanoparticle based colorimetric aptasensor platform. It was found that the colorimetric response in saliva was always suppressed owing to the matrix effects. By navigating the challenging interfering issues in saliva, we were successfully able to detect methamphetamine at nanomolar levels in saliva offering immense promise for the translation of these platforms for on-site diagnostic systems. This subsequently motivated the development of a handheld portable light meter device that can reliably transduce the aptasensors colorimetric response into absorbance, facilitating quantitative detection of analyte concentrations on-site. This is crucial due to the prevalent unreliability and sensitivity problems of the conventional drug testing kits. The fabricated light meter device response was validated against a standard UV-Vis spectrometer to confirm reliability. The portable and cost-effective handheld detector device features sensitivity comparable to the well-established UV-Vis benchtop instrument and the easy-to-use device could potentially serve as a prototype for a commercial device in the future.

Keywords: aptasensors, colorimetric gold nanoparticle assay, point-of-care, oral fluid

Procedia PDF Downloads 57

Authors: Yara Mohamed Taha, Mohamed Ali El Desouky, Heba Kamal Abdel Hakim, Maha Hanafy Mahmoud

Abstract:

Hepatotoxicity due to drugs and toxic chemicals constitutes a crucial health problem nowadays. Medicinal plants are widely used recently for protecting against many liver disorders and inflammatory conditions. This study aims to evaluate hepatoprotective impact of green tea extract (GTE), rosemary extract (RE) and rosmarinic acid (RA) against hepatotoxins; ferric nitrilotriacetate (Fe-NTA) and diethylnitrosamine (DEN) in rats. Five groups of male Wistar rats were included; one control negative, while the other groups were treated intraperitoneally with DEN as 160 mg.kg-1 b.w. on 15th day and Fe-NTA as 5 mg.kg-1 b.w. on 33rd day. One of them was control positive. The other three groups were pre-administered with daily protective oral doses of either 200 mg.kg-1 b.w. of RE or 1 g.kg- 1 b.w. of GTE or 50 mg.kg-1 b.w. of RA two weeks prior to DEN exposure and continued till the end of the experimental period. The obtained data revealed a highly significant increase of MDA, 8-OHdG, DNA damage percent, a significant depletion of GSH and elevated Gr-1 protein expression in hepatocytes with liver tissue histopathological changes of rats exposed to DEN+Fe-NTA. Pre-administration of protective doses of RE, GTE and RA to DEN+Fe-NTA treated rats could normalize the altered biochemical, histopathological and immunohistochemical parameters. In conclusion, RE, GTE and RA showed a hepatoprotective effect against liver toxicity induced by DEN+Fe-NTA, with the best antioxidant and anti-inflammatory impact were for RA and GTE. Therefore, the current study declared that rosemary, green tea and products enriched with rosmarinic acid should be involved daily in diet of people who are exposed to chemicals and environmental toxins to protect themselves from hepatotoxicity.

Keywords: hepatotoxicity, diethylnitrosamine and ferric nitrilotriacetate, rosemary extract (RE), green tea extract (GTE), rosmarinic acid (RA)

Procedia PDF Downloads 91

Authors: Han Chih Feng, Yi-Cheng Hou, Jing-Huei Wu

Abstract:

Diabetes is a chronic disease. Nutrition knowledge and skills enable individuals with type 2 diabetes to optimize metabolic self-management and quality of life. This research studies the effect of nutritional education on diabetes inpatients in terms of their cognition, behavior, and attitude. The participants are inpatients diagnosed with diabetes at Taipei Tzu Chi Hospital. A total of 103 participants, 58 male, and 45 females, enrolled in the research between January 2018 and July 2018. The research evaluates cognition, behavior, and attitude level before and after nutritional education conducted by dietitians. The result shows significant improvement in actual consumption (2.5 ± 1.4 vs 3.8 ± 0.7; p<.001), diet control motivation (2.7 ± 0.8 vs 3.4 ± 0.6; p<.001), correct nutrition concept (1.2± 0.4 vs 2.4 ± 0.5; p<.001), learning willingness (2.7± 0.9 vs 3.4 ± 0.6; p<.001), cognitive behaviors (1.4 ± 0.5 vs 2.9 ± 0.7; p<.001). AC sugar (278.5 ± 321.5 vs 152.2 ± 49.1; p<.001) and HbA1C (10.3 ± 2.6 vs 8.6 ± 1.9; p<.001) are significant improvement after nutritional education. After nutritional education, participants oral hypoglycemic agents increased from 16 (9.2%) to 33 (19.0%), insulin decreased from 75 (43.1%) to 68 (39.1%), and hypoglycemic drugs combined with insulin decreased from 83 (47.7%) to 73 (42.0%).Further analysis shows that female inpatients have significant improvement in diet control motivation (3.91 ± 0.85 vs 4.44 ± 0.59; p<0.000), correct nutrition concept (3.24± 0.48 vs 4.47± 0.51; p<0.000), learning willingness (3.89 ± 0.86 vs 4.44 ± 0.59; p<0.000) and cognitive behaviors (2.42 ± 0.58 vs 4.02 ± 0.69; p<0.000); male inpatients have significant improvement in actual food intake (4.41± 0.92 vs 3.97 ± 0.42; p<0.000), diet control motivation (3.62 ± 0.86 vs 4.29 ± 0.62; p<0.000), correct nutrition concept (3.26 ± 0.44 vs 4.36 ± 0.49; p<0.000), learning willingness (3.72± 0.93 vs 4.33± 0.63; p<0.000) and cognitive behaviors (2.45± 0.54 vs 4.03± 0.77; p<0.000). In conclusion, nutritional education proves effective, regardless of gender, in improving an inpatient’s cognition, behavior, and attitude toward diabetes self-management.

Keywords: diabetes, nutrition education, actual consumption, diet control motivation, nutrition concept, learning willingness, cognitive behaviors

Procedia PDF Downloads 86

Authors: Fedili Benamar, Beloulou Mohamed Lamine, Ouahes Hassane, Ghattas Samir

Abstract:

Background and aims: While in Western countries, unilateral spinal anesthesia has been widely practiced for a long time, it remains little known in the local anesthesia community, and has not been the object of many studies. However, it is a simple, practical and effective technique. Our objective was to evaluate this practice in emergency anesthesia management in frail patients and to compare it with conventional spinal anesthesia. Methods: This is a prospective, observational, comparative study between hypobaric unilateral and conventional spinal anaesthesia for hip fracture surgery carried out in the operating room of the university military hospital of Staoueli. The work was spread over of 12-month period from 2019 to 2020. The parameters analyzed were hemodynamic variations, vasopressor use, block efficiency, postoperative adverse events, and postoperative morphine consumption. Results: -75 cases (mean age 72±14 years) -Group1= 41 patients (54.6%) divided into (ASA1=14.6% ASA2=60.98% ASA3=24.39%) single shoot spinal anaesthesia -Group2= 34 patients (45.3%) divided into (ASA1=2.9%, ASA2=26.4% ASA3=61.7%, ASA4=8.8%) unilateral hypobaric spinal anesthesia. -Hemodynamic variations were more severe in group 1 (51% hypotension) compared to 30% in group 2 RR=1.69 and odds ratio=2.4 -these variations were more marked in the ASA3 subgroup (group 1=70% hypotension versus group 2=30%) with an RR=2.33 and an odds ratio=5.44 -39% of group 1 required vasoactive drugs (15mg +/- 11) versus 32% of group 2 (8mg+/- 6.49) - no difference in the use of morphine in post-op. Conclusions: Within the limits of the population studied, this work demonstrates the clinical value of unilateral spinal anesthesia in ortho-trauma surgery in the frail patient.

Keywords: spinal anaesthesia, vasopressor, morphine, hypobaric unilateral spinal anesthesia, ropivacaine, hip surgery, eldery, hemodynamic

Procedia PDF Downloads 71

Authors: Gordon James Knowles

Abstract:

Historically the Chinatown district in Sandwich Islands has been plagued with the traditional vice crimes of illegal drugs, gambling, and prostitution since the early 1800s. However, a new form of psychologically addictive arcade style table gambling machines has become the dominant form of illegal revenue made in Honolulu, Hawaii. This study attempts to document the drive, desire, or will to play and wager with arcade style video gaming and understand the role of illegal game rooms in facilitating pathological gambling addiction. Indicators of police corruption by Chinese organized crime syndicates related to protection rackets, bribery, and pay-offs were revealed. Information fusion from a police science and sociological intelligence perspective indicates insurgent warfare is being waged on the streets of Honolulu by the People’s Republic of China. This state-sponsored communist terrorism in the Hawaiian Islands used “contactless” irregular warfare entailing: (1) the deployment of psychologically addictive gambling machines, (2) the distribution of the physically addictive fentanyl drug as a lethal chemical weapon, and (3) psychological warfare by circulating pro-China anti-American propaganda newspapers targeted at the small island populace.

Keywords: Chinese and Polynesian organized crime, china daily newspaper, electronic arcade style table games, gaming technology addiction, illegal compulsive gambling, and police intelligence

Procedia PDF Downloads 73

Authors: Venuste Kayinamura, V. Iyamuremye, A. Ngirabakunzi

Abstract:

Anti-retroviral therapy (ART) for HIV patient can cause a deficiency in glucose metabolism by promoting insulin resistance, glucose intolerance, and diabetes, diabetes mellitus keep increasing among HIV-infected patients worldwide but there is limited data on levels of blood glucose and its relationship with antiretroviral drugs (ARVs) and HIV-infection worldwide, particularly in Rwanda. A convenient sampling strategy was used in this study and it involved 323 HIV patients (n=323). Patients who are HIV positive under ARVs were involved in this study. The patient’s blood glucose was analyzed using an automated machine or glucometer (COBAS C 311). Data were analyzed using Microsoft Excel and SPSS V. 20.0 and presented in percentages. The highest diabetes mellitus prevalence was 93.33 % in people aged >40 years while the lowest diabetes mellitus prevalence was 6.67% in people aged between 21-and 40 years. The P-value was (0.021). Thus, there is a significant association between age and diabetes occurrence. The highest diabetes mellitus prevalence was 28.2% in patients under ART treatment for more than 10 years, 16.7% were <5years while 20% of patients were on ART treatment between 5-10 years. The P-value here is (0.03), thus the incidence of diabetes is associated with long-term ART use in HIV-infected patients. This study assessed the prevalence of diabetes among HIV-infected patients under ARVs attending the University Teaching Hospital of Butare (CHUB), it shows that the prevalence of diabetes is high in HIV-infected patients under ARTs. This study found no significant relationship between gender and diabetes mellitus growth. Therefore, regular assessment of diabetes mellitus especially among HIV-infected patients under ARVs is highly recommended to control other health issues caused by diabetes mellitus.

Keywords: anti-retroviral, diabetes mellitus, antiretroviral therapy, human immune deficiency virus

Procedia PDF Downloads 112

Authors: Peng Hou

Abstract:

Natural products produced from marine bacteria serve as an immense reservoir for anti-infective drugs and therapeutic agents. Nowadays, heterologous expression of gene clusters of interests has been widely adopted as an effective strategy for natural product discovery. Briefly, the heterologous expression flowchart would be: biosynthetic gene cluster identification, pathway construction and expression, and product detection. However, gene cluster capture using traditional Transformation-associated recombination (TAR) protocol is low-efficient (0.5% positive colony rate). To make things worse, most of these putative new natural products are only predicted by bioinformatics analysis such as antiSMASH, and their corresponding natural products biosynthetic pathways are either not expressed or expressed at very low levels under laboratory conditions. Those setbacks have inspired us to focus on seeking new technologies to efficiently edit and refractor of biosynthetic gene clusters. Recently, two cutting-edge techniques have attracted our attention - the CRISPR-Cas9 and Gibson Assembly. By now, we have tried to pretreat Brevibacillus laterosporus strain genomic DNA with CRISPR-Cas9 nucleases that specifically generated breaks near the gene cluster of interest. This trial resulted in an increase in the efficiency of gene cluster capture (9%). Moreover, using Gibson Assembly by adding/deleting certain operon and tailoring enzymes regardless of end compatibility, the silent construct (~80kb) has been successfully refactored into an active one, yielded a series of analogs expected. With the appearances of the novel molecular tools, we are confident to believe that development of a high throughput mature pipeline for DNA assembly, transformation, product isolation and identification would no longer be a daydream for marine natural product discovery.

Keywords: biosynthesis, CRISPR-Cas9, DNA assembly, refactor, TAR cloning

Procedia PDF Downloads 280