Multi Drug Research Articles

Multi Drug Resistance (MDR) Urinary Tract Infection: An Evidence Based Study

Background: Urinary tract infections are the most frequently reported infections that drive the use of antibiotics around the world. UTI is the 4th most common healthcare-associated infection. Multidrug-resistant (MDR) organisms are predominantly bacteria that are resistant to one or more classes of antimicrobials. The increasing rise in the incidence of MDR-UTI has resulted in increased morbidity, mortality, and treatment cost of the patients. Thus, it is important to highlight the magnitude of the problem, identify the risk factors that result in MDR-UTI, and to take appropriate measures to control its occurrence. Objective: To determine the magnitude of the multidrug-resistant bacteria, their antibiotic-resistant profile, andtheir effect on the treatment cost of the patients Methods: It is a descriptive study conducted in the Department of Urology at the Institute of Kidney Diseases (IKD) from Jan 2019 till 30th March 2020. A total of 54 patients with multi-drug resistant UTI were included in the study irrespective of age and gender. All the data was recorded on a structured pro-forma and was analyzed on SPSS. Results: A total of 3190 patients were operated on from Jan 2019 till 30th March 2020. Out of which 54 patients (1.6 %) developed MDR-UTI. Among them,38 were male and 16 females. The mean age of the patients was 41 ± 18.4. Urolithiasis with infections was found most frequent, in 32 (59.3%) patients. All patients were on broad-spectrum oral antibiotics and had a history of urethral catheterization before the development of MDR-UTI. The most common procedure was Emergency cystoscopy and DJ stent 15 (27.8%). Followed by Percutaneous nephrostomy in 8 (14.8%). Regarding co-morbidities, 38(68.5%) patients had none, 3 patients had diabetes and 6 patients were having Diabetes and Hypertension. Pseudomonas aeruginosa was found most frequent microorganisms in 34 (63%) patients while E.coli in 10 (18.5%) and Klebsiella in 5 (9.3%) patients. Colistin was found sensitive in 36 patients (66.7%). The mean hospital stay in MDR-UTI is 9.28± 5.17 days as compared to 2.1 days in routine cases. Approximately a 4-fold increase was observed in medicines alone in the management of MDR UTI. We recorded 1 mortality (1.9%), case of MDR urosepsis. Linear regression revealed previous use of antibiotics; catheterization, old age, and endo-urological procedures in an emergency as independent risk factors for MDR-UTI. Conclusion: MDR-uti is an emerging local problem. pseudomonas aeruginosa is the most frequently found microorganism in the present setup. it is associated with significant morbidity and very high treatment cost. Keywords: Urinary Tract Infection, Multidrug Resistance, Micro-Organism, Urology, Antimicrobials

Isolation of Multi Drug Resistant Escherichia coli from drinking water of Lahore City, Pakistan

Background: The major faecal coliform is Escherichia coli which contaminates the drinking water from human and animal faecal waste. In developing regions drinking water is a vital source of microbiological pathogens. Multi-drug resistant (MDR) is defined as resistance to one or more antibiotic groups. An E. coli count greater than 4/dl in municipal drinking water is indicative of unacceptable fecal contamination. Aim: To find out the frequency of MDR E. coli in water system of Lahore, Pakistan. Methodology: Drinking water was collected from different towns of Lahore. The samples were processed using Membrane Filtration Technique. In the present study, Multidrug Resistant E.coli was isolated and antibiotic resistant pattern was seen against 16 most commonly antimicrobials, using Kirby Bauer disc diffusion method. Results: Total 100 water samples were collected, frequency of Escherichia coliisolated was 27% and frequency of MDR E.coli was 19%. The highest resistance showed by the organism towards Ampicillin (AMP) 21(81.48%), Augmentin (AMC), and Ceftazidime (CAZ) 14(51.85%) respectively. In this study also frequency of Extended Spectrum β-Lactamases were seen. Most common organisms isolated were E. coli 7% and Klebsiella 5%. Frequency of other coliforms isolated from drinking water other than E. coli was Klebsiella species 26%, Pseudomonas species 27%, Enterobacter 7%, Citrobacter species 8% and Acinetobacter species 5%. Conclusion: This study revealed that drinking water of Lahore city is heavilycontaminated with pathogenic microorganisms. A high proportion of antibiotic resistant is due to overuse of antibiotics, in patients with mild infections and secretion of these resistant bacteria from patients to environment. One of the reasons could be the mixing of sewage lines with drinking water supply. So, there is solely requirement for further studies for the identification of the sources for these contaminants. Keywords: Isolates, E. coli, Klebsiella, Multi-drug resistant (MDR)

Isolation of Multi Drug Resistant Escherichia coli from drinking water of Lahore City, Pakistan

Background: The major faecal coliform is Escherichia coli which contaminates the drinking water from human and animal faecal waste. In developing regions drinking water is a vital source of microbiological pathogens. Multi-drug resistant (MDR) is defined as resistance to one or more antibiotic groups. An E. coli count greater than 4/dl in municipal drinking water is indicative of unacceptable fecal contamination. Aim: To find out the frequency of MDR E. coli in water system of Lahore, Pakistan. Methodology: Drinking water was collected from different towns of Lahore. The samples were processed using Membrane Filtration Technique. In the present study, Multidrug Resistant E.coli was isolated and antibiotic resistant pattern was seen against 16 most commonly antimicrobials, using Kirby Bauer disc diffusion method. Results: Total 100 water samples were collected, frequency of Escherichia coliisolated was 27% and frequency of MDR E.coli was 19%. The highest resistance showed by the organism towards Ampicillin (AMP) 21(81.48%), Augmentin (AMC), and Ceftazidime (CAZ) 14(51.85%) respectively. In this study also frequency of Extended Spectrum β-Lactamases were seen. Most common organisms isolated were E. coli 7% and Klebsiella 5%. Frequency of other coliforms isolated from drinking water other than E. coli was Klebsiella species 26%, Pseudomonas species 27%, Enterobacter 7%, Citrobacter species 8% and Acinetobacter species 5%. Conclusion: This study revealed that drinking water of Lahore city is heavilycontaminated with pathogenic microorganisms. A high proportion of antibiotic resistant is due to overuse of antibiotics, in patients with mild infections and secretion of these resistant bacteria from patients to environment. One of the reasons could be the mixing of sewage lines with drinking water supply. So, there is solely requirement for further studies for the identification of the sources for these contaminants. Keywords: Isolates, E. coli, Klebsiella, Multi-drug resistant (MDR)

Multi-Drug Resistance Genes associated with some Gram-Negative Bacteria Isolates from Shellfish in Iko and Douglas River Estuaries, in Nigeria

Background: Multi drug resistant bacterial agents that contaminate seafood cause several diseases in humans and are widely documented as a global public health challenge.
 Methods: This study evaluated the microbiological and antimicrobial resistance genes profiles of bacterial Isolates from shellfish vended at Iko and Douglas Creeks of Cross River State, Nigeria. A total of 540 shellfish (117 clams, 88 oysters, 136 periwinkles) samples were collected from various vendor at the two Creeks were analyzed. The samples were processed using standard microbiological methods to identify bacterial pathogens. Antimicrobial susceptibility was assayed using the Kirby-Bauer disk diffusion method. Isolates were screened for antimicrobial resistant genes using polymerase chain reaction.
 Results: Overall, a total of 135 bacteria isolates were identified. The most common isolate was Alcaligenes species 53(39.2%) followed by Pseudomonas species 44(32.6%), Providencia species 25(18.5%), Vibrio species 6(4.4%), and Paenalcaligenes species 7(5.2%). The isolates showed varying susceptibilities to Imipenem (36%) and amikacin (28%) but were all resistant to Trimethoprim-Sulfamethoxazole. Fifty-three isolates had a multiple antibiotic resistance index (MARI) of ≥0.9 - 1.0. Most of the bacterial isolates were detected with TEM genes (82.2%), SHV (51.8%,), VIM (50.3%) resistance genes. None of the isolates expressed Veb gene. Only 40.7% of the isolates expressed QnrB gene while none expressed QnrA and QnrS.
 Conclusion: The detection of these multidrug resistant clinically relevant bacterial species suggests a significant linkage of commonly consumed seafood in the community and environmental spread of MDR bacteria.

Prophylactic application of antibiotics selects extended‐spectrum β‐lactamase and carbapenemases producing Gram‐negative bacteria in the oral cavity

Prophylactic administration of broad-spectrum antibiotics in surgery can change the oral microbiome and induce colonization of oral cavity with Gram-negative bacteria including multidrug (MDR) or extensively drug resistant (XDR) organisms which can lead to lower respiratory tract infections. The aim of the study was to analyse the Gram-negative isolates obtained from oral cavity of the mechanically ventilated patients in ICUs, after prophylactic application of antibiotics and their resistance mechanisms and to compare them with the isolates obtained from tracheal aspirates from the same patients. The antibiotic susceptibility was determined by broth dilution method. PCR was applied to detect genes encoding β-lactamases. Marked diversity of Gram-negative bacteria and resistance mechanisms was found. High resistance rates and high rate of blaCTX-M and carbapenemase encoding genes (blaVIM-1 , blaOXA-48 ) were found among Klebsiella pneumoniae. Pseudomonas aeruginosa was found to harbour blaVIM and in one strain blaPER-1 gene, whereas Acinetobacter baumannii produced OXA-23-like and OXA-24/40-like oxacillinases and was XDR in all except one case. All XDR isolates belong to international clonal lineage II (IC II). The main finding of the study is that the prophlylactic application of antibiotics in surgery intensive care units (ICUs) is associated with the colonization of oral cavity and lower respiratory tract with Gram-negative bacteria. The identity of Gram-negative bacteria in oral cavity reflected those found in endotracheal aspirates leading to conclusion that oral swab as non-invasive specimen can predict the colonization of lower respiratory tract with resistant Gram-negative organisms and the risk for development of ventilator-associated pneumonia.

ROLE OF MEDICINAL PLANT EXTRACTS IN ERADICATION OF MULTI DRUG RESISTANT MICROBES

The Indian Ayurvedic medicine is a classical medicinal method that has a rich history and solid foundation. Herbal medicines play a significant role in the prevention and management of human diseases. For millennia, plants have been used as folk medicine. Medicinal plants are an abundant source of the secondary metabolite formed by plants, the bioactive phytochemicals. Scientific studies in recent decades have demonstrated the significant role of these phytochemicals in the prevention of infectious disease. In recent decades, the re-discovery of medicative plants as a source of potential therapies has become a great deal of interest. Plant-based product lines can regulate microbial development in different scenarios. The worldwide danger to public health arose from the resistance among distinct microbial species to multiple antimicrobial drugs. Approximately every competent infective agent has high levels of multi drug resistance with strengthened morbidity & mortality, which makes them called 'superbugs.' Only by designing entirely new methods, the multi-drug resistance crisis can be tackled. The goal of this study is true to clarify the information of medicinal herbs as a potential basis for herbal supplements against microbes with multi-drug resistance.

Multidrug resistant superbugs in pyogenic infections: a study from Western Rajasthan, India.

Introductionthe crude mortality rate due to infectious diseases in India is approximately 417 per one lakh persons and pyogenic infections are one of the significant contributor. Poor antimicrobial stewardship in India has led to an increase in multidrug resistant superbugs in both community as well as hospital settings. The aim of this study was to identify the bacterial etiology of pyogenic infections and to evaluate their antimicrobial resistance profile.Methodsthis is a retrospective observational study from January, 2018 to December, 2018. A total 1851 samples, collected as a part of patient care were included in this study. Specimens were subjected to culture on Blood agar and MacConkey agar and incubated at 37°C for 48 hours. Species identification was done as per standard laboratory protocol. Antimicrobial susceptibility testing was performed using Kirby-Bauer disc diffusion according to Clinical and Laboratory Standards Institute guidelines.Resultsof total 1851 samples, culture was positive in 61.54%. A total 70.59%, Gram negative organisms were isolated followed by Gram positive cocci in 45.48%, yeast in 1.05%, coryneform bacteria in 0.79% and in one case, non-tubercular mycobacteria was isolated. Staphylococcus aureus (30.9%) was the predominant organism isolated. Most common multi drug resistant isolates were Klebsiella spp. (74.79%) and Acinetobacter spp. (74.32%).Conclusionthis study gives an insight about the prevalence and common etiology of pyogenic infections along with their antimicrobial resistance profile in north western region of India. This study will contribute in formulating antibiotic stewardship program by selecting the antibiograms of pyogenic isolates.

Metabolic Acidosis in Multi Drug Poisoning with Antidepressants and Antipsychotics.

Objective:In this study, we evaluated the predictive factors for the occurrence of metabolic acidosis in patients with multi-drug poisoning, including antidepressants (Ad)/antipsychotics (Ap).Methods:This cross-sectional study was carried out in the referral poisoning center, Khorshid University Hospital, affiliated with Isfahan University of Medical Sciences. All patients with multi-drug ingestion, including Ad/Ap, were included in the study. Patients were divided into two groups with and without metabolic acidosis. Demographic factors, time from ingestion to admission, clinical manifestations, length of hospital stay, and outcome were compared in two groups. Binary logistic regression was used to identify factors associated with the risk of metabolic acidosis occurrence.Findings:Among the 206 evaluated patients, 45 patients (21.8%) had metabolic acidosis whom the majority were female (73.3%) with intentional purposes (77.8%). 31.1% of the patients with metabolic acidosis had tachycardia on admission (P = 0.03). Among all variables, time from ingestion to admission (P = 0.02) and lengths of hospital stay (P = 0.002) were significantly different between patients with and without metabolic acidosis. Tachycardia on admission (adjusted odds ratio [OR], 2.24; 95% confidence interval [CI]: 1.05–4.76; P = 0.036) and time from ingestion to admission (adjusted OR, 1.06; 95% CI: 1.00–1.13; P = 0.04) were also the predictive factors in occurrence of metabolic acidosis. Most of the patients survived without any complications (94.6%), of whom 72.3% had no previous underlying somatic diseases (P = 0.05).Conclusion:Admission tachycardia and the time elapsed from ingestion to admission may be considered important factors for predicting metabolic acidosis in multi-drug poisoning, including Ad/Ap.

Multi-Drug Resistance Genes associated with some Gram-Negative Bacterial Isolated from Shellfish in Iko and Douglas River Estuaries, in Nigeria

Background: Multi drug resistant bacterial agents that contaminate seafood cause several diseases in humans and are widely documented as a global public health challenge. Methods: This study evaluated the microbiological and antimicrobial resistance genes profiles of bacterial Isolates from shellfish vended at Iko and Douglas Creeks of Cross River State, Nigeria. A total of 540 shellfish (117 clams, 88 oysters, 136 periwinkles) samples were collected from various vendor at the two Creeks were analyzed. The samples were processed using standard microbiological methods to identify bacterial pathogens. Antimicrobial susceptibility was assayed using the Kirby-Bauer disk diffusion method. Isolates were screened for antimicrobial resistant genes using polymerase chain reaction. Results: Overall, a total of 135 bacteria isolates were identified. The most common isolate was Alcaligenes species 53(39.2%) followed by Pseudomonas species 44(32.6%), Providencia species 25(18.5%), Vibrio species 6(4.4%), and Paenalcaligenes species 7(5.2%). The isolates showed varying susceptibilities to Imipenem (36%) and amikacin (28%) but were all resistant to Trimethoprim-Sulfamethoxazole. Fifty-three isolates had a multiple antibiotic resistance index (MARI) of ≥0.9 - 1.0. Most of the bacterial isolates were detected with TEM genes (82.2%), SHV (51.8%,), VIM (50.3%) resistance genes. None of the isolates expressed Veb gene. Only 40.7% of the isolates expressed QnrB gene while none expressed QnrA and QnrS. Conclusion: The detection of these multidrug resistant clinically relevant bacterial species suggests a significant linkage of commonly consumed seafood in the community and environmental spread of MDR bacteria.

Nano-formulations for Different Treatment Modalities to Overcome Multi Drug Resistance in Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is one of the most common malignancies and it is the fourth cause of cancer-related mortality. The treatment of HCC with traditional chemotherapeutic agents such as doxorubicin has been limited due to poor efficacy, acquired resistance of the tumors and systemic toxicity. Verapamil as a P-gp inhibitor has demonstrated anti-cancer activities both in vitro and in vivo. Natural products and plant-derived compounds, such as ginger extract, have been investigated as safe and effective anti-cancer agents. Combination therapy offers preferable prognosis with less side effects. For example, the combination of chemotherapeutics with verapamil or ginger extract has been shown to exert beneficial therapeutic effects, overcome multidrug resistance and reduce side effects through modulation of different signaling pathways in tumor cells. Nano-sized formulations are designed to increase accumulation of drugs at the tumor environment and to improve solubility/bioavailability, promote retention, specificity, efficacy and safety. This review summarizes how to overcome cardiotoxicity and multidrug resistance evolved by doxorubicin in HCC treatment.

1184. Activity of anti-pseudomonal antibiotics among all Pseudomonas aeruginosa (PSA) at an academic medical health system, including β-lactam, multi-drug (MDR) and extensively drug resistant (XDR) strains

BackgroundPSA with MDR and XDR are a growing threat, and appropriate initial treatment of this organism is critical. C/T is a novel antibiotic with broad gram-negative in vitro susceptibility among surveillance studies. However comprehensive susceptibility analyses of C/T among PSA clinical isolates in comparison with other anti-PSA antibiotics remain limited, since routine clinical C/T susceptibility testing is not typically performed.MethodsThis study assessed all adult inpatient cultures positive for PSA from 32 months at an academic health system. Antimicrobial susceptibility was prospectively performed using Kirby Bower disk diffusion and interpreted by BIOMIC V3 during routine clinical care. Initial susceptibility testing included C/T along with amikacin, aztreonam, cefepime, ciprofloxacin, doripenem, gentamicin, imipenem, meropenem, piperacillin/tazobactam, and tobramycin. MDR and XDR isolates were identified using established definitions. The primary outcome was to quantify C/T resistant PSA (includes intermediate and resistant strains). Secondary outcomes were to determine resistance to other anti-PSA antibiotics and to identify C/T activity among isolates with MDR, XDR and pan-β-lactam resistance (PBLR = all β-lactams except C/T).ResultsA total of 2990 PSA isolates from 2339 cultures in 1311 individual patients were collected. Most cultures were from the lung (45%), followed by urine (30%), and body fluids (10%). For the primary outcome, 121/2990 (4%) of PSA isolates were C/T resistant. All PSA blood cultures were susceptible to C/T. Table 1 summarizes in vitro activity of all anti-PSA agents evaluated. C/T had the greatest percent susceptibility across all culture locations including MDR/XDR PSA isolates with median MICs of 8 µg/mL. For PBLR strains 35/66 (53%) were susceptible to C/T with a median MIC of 8 µg/mL.Table 1 ConclusionC/T susceptibility testing during routine care over a 2.5-year period revealed 96% susceptibility among PSA. C/T showed the highest susceptibility among all anti-PSA antibiotics for all culture locations and for MDR and XDR isolates. Given the high rates of resistance to traditional anti-PSA agents, the value of new agents with high rates of in vitro susceptibility in the gram-negative armamentarium is high.DisclosuresWarren Rose, PharmD, MPH, Merck (Grant/Research Support)Paratek (Grant/Research Support) Janet Radaatz, PharmD, Merck (Employee) Laura A. Puzniak, PhD, Merck (Employee) Ryan J. Dillon, MSc, Merck & Co., Inc., (Employee)

871. Assessment of the Wide-resistant Pseudomonas aeruginosa Outbreak at a University Hospital in Brazil: Have We Lost This War?!

BackgroundResistance profile analysis in Pseudomonas aeruginosa isolates is extremely important to prevent its transmission and to detect outbreaks. Broadly resistant strains (BR) have a high mortality rate in invasive infections. By analyzing the clinical and microbiological characteristics of these infections, one can define more effective actions in a nosocomial outbreak setting in a university hospital in Brazil.MethodsFrom January to September 2019, 13 patients from the oncohematology services and intensive care unit (ICU) followed by the stewardship program of a public university hospital in Brazil had Pseudomonas aeruginosa (PsA) BR infection. Resistant multidrug (MDR) was defined as resistant to three or more antimicrobial classes. Extensively resistant (XDR) was sensitive to a maximum of two antimicrobial classes. Resistant pandrug (PDR) has been defined as resistant to all antimicrobial classes. Bacterial samples were identified by the automated VITEK®2 system (BioMérieux). The resistance pattern was defined based on the CLSI-M100 2019 criteria. Colistin sensitivity was assessed by the colistin drop test (Pasteran et al., 2018). The modified carbapenemic inactivation method (mCIM) was performed by disk diffusion.ResultsThe 13 strains of PsA were isolated from 84.6% (11/13) blood cultures and 15.4% (2/13) tracheal aspirates, being 76.9% (10/13) from the oncohematology unit and 23.1% (3/13) of the ICU. The resistance profile was 23.1% (3/13) strains PsA MDR, 61.5% (8/13) PsA XDR and 15.4% (2/13) strains resistant to all classes (PsA PDR). 69.2% (9/13) of the strains were mCIM positive, in which the therapeutic option was ceftazidime/avibactam in combination with polymyxin. Regarding the sites of infection and use of devices, 53.8% (7/13) of the patients developed the infection after the use of central venous catheter and/or mechanical ventilation. The mortality rate was 76.9% (10/13).ConclusionThe investigation of the outbreak of Pseudomonas aeruginosa highlights the importance of infectious surveillance of this pathogen with this resistance profile, to better understand the causalities, minimize its damage and reduce potential recurrence of new outbreaks.Disclosures All Authors: No reported disclosures

A study on carbapenemase producing Acinetobacter sp., and identification of OXA-51 gene in isolates from patients attending a tertiary care hospital

Introduction: Acinetobacter sp., are Gram-negative non-fermenters that causes nosocomial infections. Multi drug resistance has been evolving very rapidly in Acinetobacter sp with also a dramatic increase in carbapenem resistance. Even though there are various mechanisms prevailing for such resistance, detection of Class D beta lactamases (oxacillinases) production is cumbersome. Accurate detection is done only by genotypic methods. Aims and objectives: In order to estimate the prevalence of Acinetobacter sp., its antimicrobial susceptibility pattern, the following study was conducted in a tertiary care hospital. Organisms resistant to Carbapenems were further tested for OXA-51 gene identification by polymerase chain reaction. Materials and Methods: All the study samples received during the study period were subjected to microbiological culture method and identification panel to isolate Acinetobacter sp., Antimicrobial susceptibility testing was done according to CLSI guidelines. Organisms that fit the criteria of multidrug resistant were further analysed for Carbapenem resistance by disc diffusion test and those isolates were subjected to polymerase chain reaction for identifying the presence of OXA-51 gene. Results: A total of 102 (4.8%) Acinetobacter sp., isolates were obtained from a total sample size of 2127 specimens during the study period. Over all, 60.78% of isolates were multidrug resistant (MDR) and 50.98% were imipenam (Carbapenem) resistant. All the imipenam resistant isolates were belonging to A.baumanii. A total of 52 isolates subjected to PCR showed positivity for OXA-51 gene. Conclusion: This warrants the need for genotyping for all of the Acinetobacter isolates to understand the gene variations in resistance and to know whether there is a universal presence of OXA-51 gene in all of the Acinetobacter baumannii.

Development of multi-drug resistance among relapsed tuberculosis drug addicts patients in Punjab Pakistan

Background: Tuberculosis (TB) is an infectious disease, caused by Mycobacterium Tuberculosis. It primarily affects lungs with symptoms of cough, chest pain, weight loss, haemoptysis, and fever. Tobacco smoking and alcohol consumption, criminal history increases chance of relapse with increases the risk of drug resistance among patients of relapse. Our study was aimed to find a co-relation between relapse and drug abuse along with associated symptoms. Methods and materials: A prospective cross-sectional study was conducted in 2016–17 on TB relapse patients in Lahore. A deatiled questionnaire was designed to record Information. All the relapsed cases were subjected to L-J culture for drug sensitivity testing against first and second line drugs. Results: The study included 200 relapse patients, [125: non-addicts, 75: addicts (tobacco, alcohol, drugs)]. Out of addicts (75, 42.6%)addicts completed 6 months of AT treatment before relapse as compared to non-addicts (125, 34.4%). 49.3% (37) of addicts had an imprisonment history. The incidence of MDR and XDR in addicts was 22.6% (17) and 13.33% (10) in addicts while 29.6% (37) and 13.6% (17) respectively in non-addicts. Co-morbidities found in addicts and non-addicts were 49.3%(37) and 36%(45) respectively. Co-morbidities were common in order of Hypertension (20%1, 21.6%2), Hepatitis B (9.33%1, 5.66%2) and diabetes (26.67%1, 20%2) in study. 1. Addicts 2. Non-Addicts Conclusion: Drug addicts have a high potential of TB relapse with a notable trend of Multi and Extensive drug resistance. Awareness should be spread regarding this danger of addiction, to help reduce prevalence of TB.

Sensitizing multi drug resistant Staphylococcus aureus isolated from surgical site infections to antimicrobials by efflux pump inhibitors.

Staphylococcus aureus is a common hospital acquired infections pathogen. Multidrug-resistant Methicillin-resistant Staphylococcus aureus represents a major problem in Egyptian hospitals. The over-expression of efflux pumps is a main cause of multidrug resistance. The discovery of efflux pump inhibitors may help fight multidrug resistance by sensitizing bacteria to antibiotics. This study aimed to investigate the role of efflux pumps in multidrug resistance. Twenty multidrug resistant S. aureus isolates were selected. Efflux pumps were screened by ethidium bromide agar cartwheel method and polymerase chain reaction. The efflux pump inhibition by seven agents was tested by ethidium bromide agar cartwheel method and the effect on sensitivity to selected antimicrobials was investigated by broth microdilution method. Seventy percent of isolates showed strong efflux activity, while 30% showed intermediate activity. The efflux genes mdeA, norB, norC, norA and sepA were found to play the major role in efflux, while genes mepA, smr and qacA/B had a minor role. Verapamil and metformin showed significant efflux inhibition and increased the sensitivity to tested antimicrobials, while vildagliptin, atorvastatin, domperidone, mebeverine and nifuroxazide showed no effect. Efflux pumps are involved in multidrug resistance in Staphylococcus aureus. Efflux pump inhibitors could increase the sensitivity to antimicrobials.

Comprehensive characterization of the ALMT and MATE families on Populus trichocarpa and gene co-expression network analysis of its members during aluminium toxicity and phosphate starvation stresses.

Aluminium (Al) toxicity and phosphate deficit on soils are some of the main problems of modern agriculture and are usually associated. Some plants are able to overcome these stresses through exuding organic acids on the rhizosphere, such as citrate and malate, which are exported by MATE (Multi drug and toxin extrusion) and ALMT (Aluminium-activated malate transporter) transporters, respectively. Despite its co-action on acidic soils, few studies explore these two families' correlation, especially on tree crops, therefore we performed a comprehensive description of MATE and ALMT families on Populus trichocarpa as a model species for arboreal plants. We found 20 and 56 putative members of ALMT and MATE families, respectively. Then, a gene co-expression network analysis was performed using broad transcriptomic data to analyze which members of each family were transcriptionally associated. Four independent networks were generated, one of which is composed of members putatively related to phosphate starvation and aluminum toxicity stresses. The PoptrALMT10 and PoptrMATE54 genes were selected from this network for a deeper analysis, which revealed that in roots under phosphate starvation stress the two genes have independent transcriptional profiles, however, on the aluminum toxicity stress they share some common correlations with other genes. The data presented here help on the description of these gene families, of which some members are potentially involved in plant responses to acid soil-related stresses and its exploration is an important step towards using this knowledge on breeding programs for P. trichocarpa and other tree crops.

Effect of ABCB1 Multidrug Resistance Protein on Efficacy of Anti-Myeloma Drugs in Carfilzomib-Resistant Myeloma Model

Development of multidrug resistance is a major problem in treating cancer patients. Over-expression of a multidrug resistance protein MDR/P-gp/ABCB1 is recognized as a mediator of resistance to chemotherapy drugs in many cancers, including hematological malignancies. Recent studies have demonstrated that ABCB1 mediates resistance to a proteasome inhibitor carfilzomib. In this study we employ an isogenic ABCB1+ and ABCB1- pair of myeloma cell lines to investigate the effect of ABCB1 expression on a broader panel of anti-myeloma drugs. Expression of ABCB1 gene was assessed in publicly available gene expression datasets, including patient samples and murine models. ABCB1+ carfilzomib-resistant subline of myeloma cells AMO-1 was subject to CRISPR/Cas knock-out of ABCB1 gene. Two clones were isolated as a result of this procedure: one clone is ABCB1-negative and the other still bearing ABCB1. A panel of myeloma drugs consisting of carfilzomib, bortezomib, melflufen (melphalan flufenamide), melphalan, bendamustine, pomalidomide, iberdomide, panobinostat, doxorubicin, daunorubicin, vincristine, and paclitaxel was investigated in cytotoxicity assays using ABCB1+ and ABCB1- isogenic sublines of carfilzomib-adapted derivative of AMO-1. Myeloma multi-drug resistance (MMDR) score for each drug was calculated as a ratio between IC50 in ABCB1+ subline to IC50 in a ABCB1- subline. Apoptosis was measured by flow cytometry. According to the regression analysis of GSE136337, elevated expression of ABCB1 mRNA is associated with shorter PFS and OS in MM patients (Figure 1, A and B). Up-regulated murine Abcb1b expression was found in murine Vk*MYC myeloma tumors (GSE111921) prone to development of extramedullary disease (EMD) (Figure 1C). Cytotoxicity assays revealed a group of myeloma drugs vulnerable to elevated ABCB1 expression including carfilzomib, panobinostat, doxorubicin, daunorubicin, vincristine and paclitaxel (Figure 1D). No cytotoxicity against carfilzomib-resistant derivatives of AMO-1 myeloma cell line was observed for pomalidomide and iberdomide. Cytotoxicities of melflufen, bortezomib, melphalan, and bendamustine were only mildly affected by elevated ABCB1 expression (Figure 1D). In line with this observation, apoptosis upon carfilzomib treatment was reduced in ABCB1+ myeloma cells, whereas treatment with melflufen has resulted in equal cell death rates in both ABCB1- and ABCB1+ clones (Figure 1E). ABCB1 efflux pump is well recognized as a mediator of multi drug resistance in cancer. In this study we investigated the effect of ABCB1 on the efficacy of myeloma drugs in carfilzomib-resistant myeloma cells. In vitro efficacies of carfilzomib, panobinostat, doxorubicin, vincristine, and paclitaxel are negatively affected by ABCB1 over-expression (MMDR>4), whereas melflufen, bortezomib, melphalan, and bendamustine were not greatly affected (MMDR<2) (Figure 1F). However, melflufen has shown better cytotoxicity in carfilzomib-resistant myeloma, which is 20-fold greater compared to melphalan and 200-fold greater compared to bendamustine (Figure 1D). In addition, we found elevated ABCB1 expression in murine myeloma tumors prone to develop EMD. Moreover, elevated ABCB1 mRNA expression was associated with shorter PFS and OS in patients with multiple myeloma. Thus, this study may propose a novel therapy strategy including a novel lipophilic peptide-drug conjugate melflufen for myeloma patients with elevated ABCB1 which may potentially develop in carfilzomib-relapsed/refractory setting or EMD. Figure 1 Disclosures Byrgazov: Oncopeptides AB: Current Employment. Slipicevic:Oncopeptides AB: Current Employment. Lehmann:Oncopeptides AB: Current Employment, Current equity holder in publicly-traded company.

Antibacterial activity of lactic acid bacteria isolated from milk and traditional fermented dairy products of south Algeria against multidrug resistance pathogenic bacteria

The alarming situation of the increasing problem of the antibiotics resistance in the whole world requires an immediate need of the exploration of better alternatives for combating resistance. The main aim of the present work was the isolation and the screening of large range of lactic acid bacteria with important potential of antibacterial activity against multidrug resistance pathogenic strains and the characterization of the molecules responsible for anti-bacterial activity. The primary screening indicated the isolation of total of 200 lactic acid bacteria (LAB) from harvested milk (camel and goat) and tradi-tional fermented dairy products (Jben and kamaria) of south Algeria and in-vestigated for their antibacterial activity against clinical pathogens bacteria. Furthermore, Nineteen representative isolates (19) were selected and char-acterized with regard to their functional properties. The obtained results indicated that produced bacteriocin by both bacterial strains L. raffinolactis LAB9 and Leuconostoc mesenteroides subsp. cremoris LAB13 has manifested an excellent antagonistic activity against P.aeruginosa (PA) and Staphylococcus aureus (MRSA), which was Methicillin resistant Staphylococcus aureus (MRSA). A Further investigation indicated that the produced bacteriocin by LAB9 and LAB 13 were heat stable, where their inactivation was observed when treated with pepsin and trypsin. Additionally, the purification of the produced bacteriocin by L.raffinolactis LAB9 and Leuconostoc mesenteroides subsp. cremoris LAB13 has been achieved by the using of the partial ammonium sulfate precipitation. The technological properties and the stability of bacteriocins of LAB may lead for bioprospection of antibacterial components in the current struggle against increasing pandrug resistance and slowing down the expansion of multi drug resistance.

Diagnostic value of matrix metalloproteinase-2 and high mobility group box 1 in patients with refractory epilepsy

IntroductionThere are large numbers of inflammatory molecules and humoral mediators that can be involved in the epileptogenesis such as cytokines, matrix metalloproteinases (MMP), and high mobility group box-1 (HMGB1). We aimed to evaluate serum levels and the diagnostic value of MMP-2 and HMGB1 in Iraqi patients with epilepsy.MethodsOne hundred epileptic patients comprised 60 controlled epileptics and 40 refractory patients to treatment with multi antiepileptic drugs (AEDs). Other 50 family-unrelated age- and sex-matched healthy subjects were selected to represent the control group. Serum levels of MMP-2 and HMGB1 were estimated using ELISA. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of these markers when required.ResultsMMP-2 level was significantly higher in controls than epileptic patients in general (controlled and refractory patients). ROC curve, showed poor diagnostic value of MMP-2 in discriminating epileptics into responsive or refractory to treatment from controls (AUC = 0.679 (95% CI = 0.536-0.823), and AUC = 0.77 (95% CI = 0.637-902), respectively). Serum HMGB1 level in epileptic patients and controls was in close approximation to each other.ConclusionsMMP-2 is significantly decreased in patients particularly those with refractory epilepsy (RE); however, it has poor diagnostic value. No difference in the serum HMGB1 level between epileptic patients and controls.

Molecular and cellular paradigms of multidrug resistance in cancer.

The acquisition of resistance to chemotherapy is a major hurdle in the successful application of cancer therapy. Several anticancer approaches, including chemotherapies, radiotherapy, surgery and targeted therapies are being employed for the treatment of cancer. However, cancer cells reprogram themselves in multiple ways to evade the effect of these therapies, and over a period of time, the drug becomes inactive due to the development of multi-drug resistance (MDR). MDR is a complex phenomenon where malignant cells become insensitive to anticancer drugs and attain the ability to survive even after several exposures of anticancer drugs. In this review, we have discussed the molecular and cellular paradigms of multidrug resistance in cancer. An Extensive research in cancer biology revealed that drug resistance in cancer is the result of perpetuated intracellular and extracellular mechanisms such as drug efflux, drug inactivation, drug target alteration, oncogenic mutations, altered DNA damage repair mechanism, inhibition of programmed cell death signaling, metabolic reprogramming, epithelial mesenchymal transition (EMT), inherent cell heterogeneity, epigenetic changes, redox imbalance, or any combination of these mechanisms. An inevitable cross-link between inflammation and drug resistance has been discussed. This review provided insight molecular mechanism to understand the vulnerabilities of cancer cells to develop drug resistance. MDR is an outcome of interplays between multiple intricate pathways responsible for the inactivation of drug and development of resistance. MDR is a major obstacle in regimens of successful application of anti-cancer therapy. An improved understanding of the molecular mechanism of multi drug resistance and cellular reprogramming can provide a promising opportunity to combat drug resistance in cancer and intensify anti-cancer therapy for the upcoming future.