Broad Drug Resistance Research Articles

Long-term results of treatment of patients with newly detected pulmonary tuberculosis

Introduction. The article is devoted to the effectiveness of the treatment of pulmonary tuberculosis with multiple (MDR) and broad drug resistance (XDR) M. tuberculosis (MBT) in patients after the end of the main course of treatment (intensive and continuation phase), which were observed with clinical cure in 3rd dispensary registration group.Aim. To study the long-term results of treatment of newly diagnosed patients with pulmonary tuberculosis with MDR and XDR MBT with the determination of significant risk factors for relapse in a megalopolis.Materials and methods. The main course of treatment was completed by 119 newly diagnosed patients with pulmonary tuberculosis with MDR or XDR TB for the period from 2013 to 2019, who were treated in anti-tuberculosis hospitals in St. Petersburg and outpatient in St. Petersburg Interdistrict Petrograd-Primorsky TB Dispensary No. 3. The patients were divided into two groups: the main group (MG) consisted of 40 people who received treatment regimens that included drugs (thioureidoiminomethylpyridinium perchlorate (perchlosone, bedaquiline, linezolid ), the control group (CG) – 79 people who received who received standard therapy, without the above drugs. The groups are comparable in clinical, radiological and bacteriological characteristics. The course of treatment was found effective in 86 (72.3%): in MG – in 34 (85.0%) people, in CG – in 52 (65.8%) people, p < 0.01. These 86 patients were transferred to the 3rd group of dispensary registration with a diagnosis of “clinical cure”.Results. Adverse treatment outcomes were analyzed, including the causes of tuberculosis recurrence. The use of drugs (perchlosone, bedaquiline, linezolid) in the complex chemotherapy of tuberculosis with MDR and XDR MBT increases the effectiveness of treatment of patients and significantly less often they have a recurrence of pulmonary tuberculosis (12.5%), compared with the group without the use of these drugs (27.8%). The most significant risk factors for relapses of drug-resistant tuberculosis: the preservation of residual cavities in the absence of bacterial excretion at the end of the main c male gender, concomitant pathology (HIV infection), past incarceration, late detection of the disease and destructive forms of tuberculosis.Conclusions. The main course of therapy was found to be effective in 86 (72.3%) in patients with MG – in 34 (85.0%) people, in CG – in 52 (65.8%) people, p < 0.01. All the patients who completed the course of therapy were transferred to the 3rd group of dispensary registration with a diagnosis of “clinical cure”. Relapses of the disease occurred much less frequently in patients receiving perchlozon, bedaquiline, linezolid in complex therapy compared with the control group.

Comparative genomics and DNA methylation analysis of Pseudomonas aeruginosa clinical isolate PA3 by single-molecule real-time sequencing reveals new targets for antimicrobials.

Pseudomonas aeruginosa (P.aeruginosa) is an important opportunistic pathogen with broad environmental adaptability and complex drug resistance. Single-molecule real-time (SMRT) sequencing technique has longer read-length sequences, more accuracy, and the ability to identify epigenetic DNA alterations. This study applied SMRT technology to sequence a clinical strain P. aeruginosa PA3 to obtain its genome sequence and methylation modification information. Genomic, comparative, pan-genomic, and epigenetic analyses of PA3 were conducted. General genome annotations of PA3 were discovered, as well as information about virulence factors, regulatory proteins (RPs), secreted proteins, type II toxin-antitoxin (TA) pairs, and genomic islands. A genome-wide comparison revealed that PA3 was comparable to other P. aeruginosa strains in terms of identity, but varied in areas of horizontal gene transfer (HGT). Phylogenetic analysis showed that PA3 was closely related to P. aeruginosa 60503 and P. aeruginosa 8380. P. aeruginosa's pan-genome consists of a core genome of roughly 4,300 genes and an accessory genome of at least 5,500 genes. The results of the epigenetic analysis identified one main methylation sites, N6-methyladenosine (m6A) and 1 motif (CATNNNNNNNTCCT/AGGANNNNNNNATG). 16 meaningful methylated sites were picked. Among these, purH, phaZ, and lexA are of great significance playing an important role in the drug resistance and biological environment adaptability of PA3, and the targeting of these genes may benefit further antibacterial studies. This study provided a detailed visualization and DNA methylation information of the PA3 genome and set a foundation for subsequent research into the molecular mechanism of DNA methyltransferase-controlled P. aeruginosa pathogenicity.

WR1065 conjugated to thiol-PEG polymers as novel anticancer prodrugs: broad spectrum efficacy, synergism, and drug resistance reversal.

The lack of anticancer agents that overcome innate/acquired drug resistance is the single biggest barrier to achieving a durable complete response to cancer therapy. To address this issue, a new drug family was developed for intracellular delivery of the bioactive aminothiol WR1065 by conjugating it to discrete thiol-PEG polymers: 4-star-PEG-S-S-WR1065 (4SP65) delivers four WR1065s/molecule and m-PEG6-S-S-WR1065 (1LP65) delivers one. Infrequently, WR1065 has exhibited anticancer effects when delivered via the FDA-approved cytoprotectant amifostine, which provides one WR1065/molecule extracellularly. The relative anticancer effectiveness of 4SP65, 1LP65, and amifostine was evaluated in a panel of 15 human cancer cell lines derived from seven tissues. Additional experiments assessed the capacity of 4SP65 co-treatments to potentiate the anticancer effectiveness and overcome drug resistance to cisplatin, a chemotherapeutic, or gefitinib, a tyrosine kinase inhibitor (TKI) targeting oncogenic EGFR mutations. The CyQUANT®-NF proliferation assay was used to assess cell viability after 48-h drug treatments, with the National Cancer Institute COMPARE methodology employed to characterize dose-response metrics. In normal human epithelial cells, 4SP65 or 1LP65 enhanced or inhibited cell growth but was not cytotoxic. In cancer cell lines, 4SP65 and 1LP65 induced dose-dependent cytostasis and cytolysis achieving 99% cell death at drug concentrations of 11.2 ± 1.2 µM and 126 ± 15.8 µM, respectively. Amifostine had limited cytostatic effects in 11/14 cancer cell lines and no cytolytic effects. Binary pairs of 4SP65 plus cisplatin or gefitinib increased the efficacy of each partner drug and surmounted resistance to cytolysis by cisplatin and gefitinib in relevant cancer cell lines. 4SP65 and 1LP65 were significantly more effective against TP53-mutant than TP53-wild-type cell lines, consistent with WR1065-mediated reactivation of mutant p53. Thus, 4SP65 and 1LP65 represent a unique prodrug family for innovative applications as broad-spectrum anticancer agents that target p53 and synergize with a chemotherapeutic and an EGFR-TKI to prevent or overcome drug resistance.

Efficacy of a shorter chemotherapy regimen in a patient with drug-resistant tuberculosis with early surgical intervention

Introduction. Treatment for multidrug-resistant tuberculosis lasts 18 to 24 months, so there is a high rate of therapy interruptions, including patient-initiated interruptions. In recent years, the use of shorter regimens for the treatment of multidrug-resistant tuberculosis has been actively discussed around the world. The first use of shorter chemotherapy regimens was conducted in three regions of the Russian Federation in 2019. However, the prevalence of the use of shorter chemotherapy regimens in our country remains low.The purpose of the study was to demonstrate the high efficacy of a shortened chemotherapy regimen with surgical intervention.Materials and methods. In the conditions of the Central Research Institute of Tuberculosis, the patient underwent a comprehensive examination, including mandatory, additional and optional methods of investigation. Based on the findings of the comprehensive examination and established broad drug resistance of Mycobacterium tuberculosis, a shorter course of chemotherapy was prescribed and surgical treatment was performed after 2 months of therapy.Results and Discussion It was decided to carry out anti-tuberculosis therapy according to a shortened scheme in the amounts: Linezolid, Bedaquiline, Cycloserine, Levofloxacin, Pyrazinamide. Taking into account positive radiological dynamics after two months of treatment and absence of contraindications, early surgery intervention was performed. At the end of a nine-month course of chemotherapy the patient had a stable radiological picture, “fresh” focal and infiltrative changes in the lungs were not detected. Early surgical treatment with a shorter chemotherapy regimen may become a highly effective method in the cure of drug-resistant tuberculosis in the Russian Federation.Conclusion. A shorter chemotherapy regimen with optimal timing of surgical intervention is highly effective and safe to use.

Resistance and virulence gene analysis and molecular typing of Escherichia coli from duck farms in Zhanjiang, China.

The widespread use of antibiotics in animal agriculture has increased the resistance of Escherichia coli, and pathogenic E. coli often harbor complex virulence factors. Antimicrobial resistance in pathogenic bacteria can cause public health problems. Correlation analyses of the resistance, virulence, and serotype data from the pathogenic bacteria found on farms and in the surrounding environment can thus provide extremely valuable data to help improve public health management. In this investigation, we have assessed the drug resistance and virulence genes as well as the molecular typing characteristics of 30 E. coli strains isolated from duck farms in the Zhanjiang area of China. Polymerase chain reaction was used to detect the drug resistance and virulence genes as well as serotypes, and whole-genome sequencing was used to analyze the multilocus sequence typing. The detection rates for the oqxA resistance gene and fimC virulence gene were highest (93.3%, respectively). There were no correlations between the drug resistance and virulence gene numbers in the same strain. The epidemic serotype was O81 (5/24), ST3856 was an epidemic sequence type, and strains I-9 and III-6 carried 11 virulence genes. The E. coli strains from the duck farms in the Zhanjiang area were thus found to have a broad drug resistance spectrum, various virulence genes, complex serotypes, and certain pathogenicity and genetic relationship. Monitoring the spread of pathogenic bacteria and the provision of guidance regarding the use of antibiotics in the livestock and poultry industries will be required in the future in the Zhanjiang area.

Investigation of plant metabolites as potential inhibitors of Acinetobacter baumannii: An In-Silico approach

The gram-negative bacteria Acinetobacter baumannii is responsible for a broad spectrum of dangerous nosocomial infections. Acinetobacter baumannii is easily transmitted into the body via mechanical ventilators, open wounds, and intravascular catheters. Bloodstream infections and ventilator-associated pneumonia caused by A. baumannii have the greatest mortality rates. Due to its multidrug resistance (MDR), broad drug resistance, and pan-drug resistance phenotypes, A. baumannii is now recognized as a particularly challenging pathogen to control. In order to develop novel therapeutics for the existing condition, an in-silico approach was used. Based on the review of the literature, five drug target proteins including OmpA (Peptidoglycan-binding domain), CarO, DcaP, OmpW, and PBP were identified to be vital for the pathogen's survival, infection to the host, and multidrug resistance. A total of 20 potential plant metabolites with antibacterial properties were docked against these proteins. Among them, Corilagin, (+)-Lyoniresinol-3 alpha-O-beta-d-glucopyranoside, Epsilon-Viniferin, and Epigallocatechin gallate (EGCG) showed superior binding energy compared to the reference drug carbapenem. Corilagin and -(+)-Lyoniresinol-3 alpha-O-beta-d-glucopyranoside showed the best binding energy with DcaP protein having a docking score of about −261.74, and −238.19 respectively. Those two best protein-ligand complexes were assessed for MD simulation where the average RMSD value of 3.5 A° for Dcap-Corilagin and 3.0 A° for Dcap-(+)-Lyoniresinol-3 alpha-O-beta-d-glucopyranoside indicated their excellent stability. In-silico ADME analysis showed all four metabolites had a good estimated solubility (ESOL) between −3.56 and −6.32, a Log P range of 1.87–2.71 and they responded negatively in Blood-Brain Barrier, and CYP inhibition. Toxicity study showed that the expected new drugs, particularly Corilagin, are completely non-carcinogenic, non-toxic, and safe to use as therapeutic treatments against Acinetobacter baumannii. Therefore, all the predicted four metabolites could be used as a medication against Acinetobacter baumannii. As a result of the promising outcomes from our present study, we strongly recommend more in vivo analysis for experimental confirmation.

Ways to improve the effectiveness of treatment of patients with destructive pulmonary tuberculosis with multiple and broad drug resistance of the pathogen

SummaryTreatment of patients with destructive forms of drug-resistant pulmonary tuberculosis remains a difficult problem. The presence of a destruction cavity in the lung reduces the effectiveness of therapy and is a negative prognostic factor. The objective of the study: to search for the optimal tactics for the treatment of destructive drug-resistant pulmonary tuberculosis and to evaluate the impact of collapsing procedures and surgical interventions on the outcome of treatment according to immediate and long-term observations. Materials and methods. The study included 398 patients with destruc-tive pulmonary tuberculosis with registered multiple (MDR-TB) in 61.3% and extensively drug-resistant (XDR-TB) in 38.7% of patients. Depending on the treat-ment tactics chosen in the clinic, 2 groups of patients were formed who received individualized chemothera-py based on the results of drug susceptibility testing of the pathogen and were comparable in other significant evaluation criteria. The main group consisted of 269 pa-tients, whose management tactics included the staged use of invasive methods of treatment, the comparison group — 129 patients who received only adequately selected anti-tuberculosis chemotherapy due to the im-possibility of using invasive methods. Staged invasive treatment included collapsotherapeutic techniques used in the absence of regression of the tuberculosis process against the background of adequately selected chemo-therapy for 1.5–2 months, and, if the previous stage was ineffective, resection surgery 4–6 months after the start of treatment in the clinic. Results: In the main group, collapse therapy procedures were performed in 179/269 (66.5%) patients, and they were successful in 106/115 (92.2%) patients with MDR-TB and in 51/64 (79.7%) pa-tients with XDR TB (p=0.44). Surgical interventions were performed in 25/269 (9.3%) patients, which were effec-tive in 20/25 (80%). Postoperative complications were registered in 4/25 (16%) patients. In general, microbiolog-ical conversion of sputum and closure of decay cavities were registered in 245/269 (91.1%) patients of the main group and in 75 out of 129 (58.6%) of the comparison group. Conclusion. Timely correction of treatment tac-tics based on the staged application of collapse therapy and surgical methods made it possible to achieve a cure in 91.1% of previously ineffectively treated patients with destructive pulmonary MDR/XDR tuberculosis.

Detection of NDM-1-Positive Aeromonas caviae from Bacteremia by Using Whole-Genome Sequencing.

PurposeNosocomial infections caused by New Delhi metallo-β-lactamase (NDM)-producing bacteria are prevalent worldwide. However, such diseases caused by NDM-producing Aeromonas caviae had never been reported. Our study aimed to elucidate the genomic characteristics of NDM-1-producing A. caviae isolated from hospital patients.MethodsBacterial genomic features and possible origins were assessed by whole-genome sequencing (WGS) and phylogenetic analysis. Subsequent investigations include antimicrobial susceptibility testing and multilocus sequence typing (MLST).ResultsWe identified here two NDM-1-producing A. caviae isolates from bacteremia. Susceptibility testing showed that two isolates were multi-drug resistant and shared a similar resistance profile and were only sensitive to amikacin and trimethoprim/sulfamethoxazole. Both A. caviae isolates carry the carbapenem resistance gene blaNDM-1 and also have antibiotic resistance genes such as β-lactams, AmpC enzymes, macrolides, aminoglycosides, and quinolones. S1-PFGE and Southern blot analysis were negative. Whole-genome sequencing and comparative analysis revealed that these two isolates shared a close relationship.ConclusionTo the best of our knowledge, this work describes the first detection of non-plasmid encoded blaNDM-1 in A. caviae. The A. caviae isolated in this study has a broad drug resistance spectrum. Phenotypic and molecular analysis indicated the two isolates belong to the same clone. Routine genomic surveillance of this species is now necessary to effectively curb the further dissemination of carbapenem-resistant bacteria in the region.

МОЖЛИВОСТІ ТЕХНОЛОГІЇ СЕКВЕНУВАННЯ НОВОГО ПОКОЛІННЯ ДЛЯ ВИЗНАЧЕННЯ СТІЙКОСТІ ШТАМІВ М. TUBERCULOSIS ДО АНТИМІКОБАКТЕРІАЛЬНИХ ПРЕПАРАТІВ

POSSIBILITIES OF NEW GENERATION SEQUENCING TECHNOLOGY FOR THE DETERMINATION OF RESISTANCE OF M. TUBERCULOSIS STRAINS TO ANTIMYCOBACTERIAL DRUGS Y. A. Cherednik, O. A. Zhurilo, A. I. Barbova, O. V. Pavlova Abstract The aim of the study was to analyze the possibilities and informativeness of the new generation sequencing technology in the variant of targeted sequencing of the panel of genes responsible for drug resistance to a wide range of antimycobacterial drugs with Deeplex Myc-TB analytical solution for use in clinical practice. Materials and methods. M. tuberculosis DNA was isolated from 22 cultures obtained on Levenstein-Jensen medium using the QIAamp® DNA Mini Kit. The Deeplex Myc-TB kit was used to amplify the target panel. Sequencing was performed on the Illumina platform (MiSeq, NextSeq 500, Nextera XT library kit). 15 Ukrainian isolates from patients with pulmonary tuberculosis were also characterized by ETR-VNTR PCR typing methods by ETR loci (Exact Tandem Repeats) A, B, C, D and E. Results. According to the sequence data, out of 15 Ukrainian isolates (from Kyiv city population) M. tuberculosis 9 (60.0%) isolates were multidrug resistant and 4 (27.0%) isolates were characterized by broad drug resistance. All MDR and BDR strains had identical mutations, determining resistance to rifampicin (rpoB1 S450L) and isoniazid (katG S315T). Three strains additionally had common mutations that caused resistance to ethionamide (fabG C-15T), ethambutol (embB Y334H) and streptomycin (rpsL K88R). Another group of strains had common mutations in the genes, responcible for resistance to ethambutol (embB M306V) and streptomycin (rpsL K43R). This may indicate the existence of clusters of strains that are already resistant to these 5 or 4 drugs and prevalent in the population. New mutations with unknown effect on resistance have been found in genes and regions responsible for resistance to rifampicin, pyrazinamide, ethambutol, streptomycin, fluoroquinolones, kanamycin, amikacin, capreomycin, ethionamide. Conclusions. The Deeplex Myc-TB analytical solution provides a wealth of relevant information on antimycobacterial drug resistance markers and speeds up the data analysis process. To address epidemiological issues, it will be important to use additional typing methods and search for new phylogenetic markers of the most common in the population of clusters of M. tuberculosis strains. Key words: tuberculosis, mycobacteria tuberculosis, drug resistance, polymerase chain reaction, sequencing.

The Effectiveness of Autologous Composition Based on Dendritic Cells in the Treatment of Pulmonary Tuberculosis with Extremely Drug Resistance After an Ineffective Course of Polychemotherapy

Despite the introduction of new anti-tuberculosis drugs, the problem of treating tuberculosis patients with a broad drug-resistant pathogen (XDR MBT) remains relevant.Aim of the study. Тo increase the effectiveness of treatment of this category of patients using an autologous composition based on dendritic cells activated by a specific antigen.Research materials and methods. А prospective study conducted in 2016-2019 included 52 patients, 36 (69.2%) men and 16 (30.8%) women, average age 43.2 ± 1.5 years. All patients received antituberculosis therapy in accordance with existing regulatory documents and recommendations. The main group consisted of 25 patients (48.1%) who agreed to immunotherapy with the introduction of an autologous composition with activated specific peptides ESAT-6 and SFP-10 dendritic cells against the background of anti-tuberculosis therapy. Statistical processing of the material carried out using the Statistica 10.0 application software package.Results. In 70.0% of cases (n = 14), there were no adverse reactions to suspension administration in patients of the main group. One patient (7.2%) had a general reaction in the form of tremor, chills after the first injection. In 5 (38.7%) patients on the 8th and 9th administration, a skin reaction was noted at the injection sites in the form of a papule, with a final regression of these changes after 8 weeks. By 18– 24 months of the course, anti-tuberculosis therapy in combination with immunotherapy in the first month of treatment allowed to obtain a positive effect according to clinical, bacteriological and radiological data in 64% of cases. It was not possible to obtain this result during therapy at the previous stages of treatment. In the comparison group, a positive result achieved only in 45.5% of cases.Conclusion. The results of the study suggest a positive effect of complex treatment with the inclusion of immunotherapy with an autologous composition based on dendritic cells in the most severe contingent of patients with pulmonary tuberculosis with broad drug resistance of the pathogen, even with an ineffective course of chemotherapy at the previous stage in 64% of cases.

Tongue cancer in a patient with pulmonary tuberculosis. Case report

Tuberculosis and malignant neoplasms are a global threat, these diseases claim millions of lives and cause tremendous suffering to people worldwide. Tuberculosis can mask cancer and create preconditions for carcinogenesis. The article presents a clinical case of tongue cancer in a patient with pulmonary tuberculosis. Patient B.N., 45 years old, was admitted to the FSBI Novosibirsk Tuberculosis Research Institute with complaints of a "sore place" on the tongue, pain in the right half of the tongue, enlargement of the submandibular lymph nodes on the right The tongue hurts for about 1,5 years, but over the last 2 months, condition has worsened. The patient noticed an increase in the size of lesion on the tongue and increased pain, especially during meals (he can eat only after pain relief, for which he takes Nimesulide 4 pills a day). In recent days, he felt also epigastric pain for which he didnt visit a doctor. Tongue biopsy was performed, and histological analysis revealed cancer. The patient was diagnosed with fibrous-cavernous tuberculosis of the upper lobe of the right lung, infiltration and seeding phase; mycobacterium tuberculosis (+); broad drug resistance (HRSECmKmPasCsOfl); gastric ulcer (0,5 cm), exacerbation stage; chronic erosive gastritis, exacerbation stage; malignant neoplasm of the floor of the mouth on the right T3(4)N1M1. With this diagnosis, the patient was discharged for further treatment by oncologists. Physicians need to have a high alertness for cancer in patients with tuberculosis, especially those in the high-risk group. A minimum of standard diagnostic procedures will allow you to verify the diagnosis timely.

Tuberculosis behind bars or the disease is not a sentence. Clinical experience of using the drug Perchlozon® in patients with tuberculosis with multy- and extended drug resistance in the moscow penitenciary

This article contains the main statistical indicators of tuberculosis among arrested and convicted persons, holding in institutions of the penitentiary system of Moscow and Russia over the period from 2016 to 2019. On the territory of Moscow all medical measures among these persons with tuberculosis are provided in conditions of the tuberculosis pulmonary departments № 1, 2 of the Hospital of the Federal state healthcare institution Medical and sanitary part-77 of the Federal Penitentiary Service of Russia. The work describes its structure and tasks. Taking into account the specifics of the group of patients, the article focuses on persons with tuberculosis/HIV co-infection. The main orders of the Federal Penitentiary Service, regulating the treatment of patients in places of deprivation of liberty, are also highlighted. The hospital uses an individual approach to each patient, including patients with tuberculosis in combination with multiple and broad drug resistance. The article considers two clinical cases, reflects the main points of clinical and diagnostic dynamics in the treatment of patients with pulmonary tuberculosis with the drug Perchlozon®. Because of the analysis of these clinical examples it follows, that on the background of the use of the drug «Perchlozon» in the scheme of the IV chemotherapy regimen, after 6 months from starting of treatment, there is abacillation and scarring of the decay cavities at the patients. It plays an important role in returning a sick person to a healthy employable society, as well as in normalization of his psychoemotional state.

ОРГАНИЗАЦИЯ РАБОТЫ С ДЕТЬМИ И ПОДРОСТКАМИ В ОЧАГАХ ТУБЕРКУЛЕЗНОЙ ИНФЕКЦИИ

Objective: Highlighting main tendencies of organization of works with children and adolescents in the outbreaks of tuberculosis infection. Methods : Retrospective analysis of the federal and industrial statistical reports (Forms 33 and 8) in Leningrad region from 2017 to 2019 was conducted. Results: The majority of children (85.7 %) having tuberculosis got infected inside the outbreaks formed by people who had already been infected – smear-positive. In 50 % of cases in smear-positive outbreaks multiple and broad drug resistance of the pathogen was detected. In every 5 th case tuberculosis was accompanied with HIVinfection, also there was a registered death of tuberculosis patient. Tuberculosis affected children 7.7 years old on average in case the average length of the contact was 2.8 years before the moment of diagnosing the disease. Conclusions : Such negative factors as HIV coinfection and pathogen drug resistance worsen epidemiological process in case of tuberculosis and change clinical structure of the disease among children and adolescents. These factors influence the effectiveness of the treatment and tuberculosis prognosis for infection source. It is important to take them into consideration when conducting a range of prevention activities in the outbreaks of the disease.

The effectiveness of the main course of chemotherapy for drug-resistant tuberculosis

SummaryThe article is devoted to the effectiveness of treatment of tuberculosis with MDR-TB and XDR-TB mycobacteria after the main course of chemotherapy. The treatment of tuberculosis with multiple and broad MD MBT is a com-plex and urgent task. Prescribing the latest generation of drugs (thioureidoiminomethylpyridinium perchlorate, bedaquiline, linezolid) in the complex composition of tu-berculosis therapy with MDR and XDR-MBT significantly increases the effectiveness of the main course of treat-ment, compared with the control group, respectively (85.0% and 65.8%). Мaterials and methods. The main course of chemotherapy was completed by 119 patients with pulmonary tuberculosis with drug-resistant patho-gen, who were diagnosed for the first time, treated in anti-tuberculosis hospitals in St. Petersburg and on an outpatient basis in St. Petersburg “Inter-district Petro-grad-Primorsky TB dispensary No 3”. Clinical, radiological, and laboratory data are presented. All patients were iden-tified with Mycobacterium tuberculosis with multiple and broad drug resistance MBT. The patients were divided into 2 groups: the main group (OG) — 40 people, received the latest generation drugs; 79 patients (KG) — received standard therapy without the inclusion of the latest gen-eration of drugs. Half of the patients in all age groups were identified during a preventive examination (50.4%), a third when applying for complaints (31.9%), and one — when examining for contact with a patient with tubercu-losis. Infiltrative tuberculosis was most often diagnosed in 75 (63.0%), disseminated tuberculosis was detected in 20 (16.8%), and fibrocavernous pulmonary tuberculo-sis — in 24 (20.2%). The decay phase was diagnosed in 88 (73.9%) patients. Three-quarters of the studied patients had 90 comorbidities (75.6%). Results and discussion. The groups are comparable in terms of clinical, radiologi-cal, and bacteriological parameters. By the fourth month of the therapy, the cessation of bacterial excretion was significantly more often determined in patients taking bedaquiline, thioureidoiminomethylpyridinium perchlo-rate, linezolid as part of HT, compared with the group not receiving the above drugs 47.5% and 35.4%, respectively, (p<0.05); by the end of the intensive phase of treatment, the cessation of bacterial excretion occurred in OG in 92.0%, KG — 57.0% (p<0.001). Conclusions. The inclu-sion of drugs thioureidoiminomethylpyridinium perchlo-rate, bedaquiline, linezolid in the complex chemotherapy of tuberculosis with multi-resistant tuberculosis increases the effectiveness of the main course of treatment com-pared to the KG.

Clinical manifestations and outcomes of pulmonary tuberculosis in patients with different genotypes of the pathogen

Objective: to study the clinical manifestations and effec- tiveness of treatment of patients with pulmonary tubercu- losis with different genotypes of M. tuberculosis. Materi­ als and methods. Clinical and microbiological data of 120 patients were stratified on the M. tuberculosis epidemic variants: subtypes of Beijing CC2 / W148, CC1, other sub- types of Beijing, and other genotypes other than Beijing (nonBeijing). The DNA of M. tuberculosis strains obtained were genotyped by RD 105/207 for Beijing genotype iden- tification. The CC2/W148 subtype was identified by the presence of a specific deletion in the kdpD gene, the CC1 subtype was done by the SNP in the pks17 gene. Results. It was found that in tuberculosis caused by the CC2/W148 subtype of the Beijing genotype, multiple and broad drug resistance occur in 87.5% of patients. The CC2/W148 ge- no type contributes to the chronization of the tuberculosis process, the formation of fibrous-cavernous tuberculo- sis. Miliary tuberculosis is more common (37.5%) among newly diagnosed patients with CC2/W148 than in other genotypes (15.7%), which is also associated with a high incidence of concomitant HIV infection (56.3%). Total lung tissue damage in patients with infiltrative tuberculosis is observed in 41.7% of patients with the CC2/W148 MBT subtype. The clinical cure rate in patients with CC2/W148 is significantly lower (18.7% compared to 47.4%) than in tuberculosis caused by other genotypes. Conclusion. The results obtained indicate the presence of clinical features of tuberculosis caused by the w148 / CC2 subtype of the Beijing genotype, and the expediency of further study of the clinical, radiological and microbio- logical characteristics of the tuberculosis process caused by MBT of different subtypes of the Beijing genotype and non-Beijing genotypes.

Enhanced Protein Damage Clearance Induces Broad Drug Resistance in Multitype of Cancers Revealed by an Evolution Drug-Resistant Model and Genome-Wide siRNA Screening.

Resistance to therapeutic drugs occurs in virtually all types of cancers, and the tolerance to one drug frequently becomes broad therapy resistance; however, the underlying mechanism remains elusive. Combining a whole whole‐genome‐wide RNA interference screening and an evolutionary drug pressure model with MDA‐MB‐231 cells, it is found that enhanced protein damage clearance and reduced mitochondrial respiratory activity are responsible for cisplatin resistance. Screening drug‐resistant cancer cells and human patient‐derived organoids for breast and colon cancers with many anticancer drugs indicates that activation of mitochondrion protein import surveillance system enhances proteasome activity and minimizes caspase activation, leading to broad drug resistance that can be overcome by co‐treatment with a proteasome inhibitor, bortezomib. It is further demonstrated that cisplatin and bortezomib encapsulated into nanoparticle further enhance their therapeutic efficacy and alleviate side effects induced by drug combination treatment. These data demonstrate a feasibility for eliminating broad drug resistance by targeting its common mechanism to achieve effective therapy for multiple cancers.

Antibiotic susceptibility signatures identify potential antimicrobial targets in the Acinetobacter baumannii cell envelope

A unique, protective cell envelope contributes to the broad drug resistance of the nosocomial pathogen Acinetobacter baumannii. Here we use transposon insertion sequencing to identify A. baumannii mutants displaying altered susceptibility to a panel of diverse antibiotics. By examining mutants with antibiotic susceptibility profiles that parallel mutations in characterized genes, we infer the function of multiple uncharacterized envelope proteins, some of which have roles in cell division or cell elongation. Remarkably, mutations affecting a predicted cell wall hydrolase lead to alterations in lipooligosaccharide synthesis. In addition, the analysis of altered susceptibility signatures and antibiotic-induced morphology patterns allows us to predict drug synergies; for example, certain beta-lactams appear to work cooperatively due to their preferential targeting of specific cell wall assembly machineries. Our results indicate that the pathogen may be effectively inhibited by the combined targeting of multiple pathways critical for envelope growth.

Pharmacotyping pancreatic cancer patients with organoids

Background: Pancreatic cancer is a devastating disease and new, personalized approaches for treatment are much-needed. Organoids are a new promising advance on this road. We have used pancreatic cancer organoids for pharmacotyping. The aim of this pilot was to find individually best suitable drugs amongst clinically relevant alternatives. The long term aim is to improve prognosis and decrease the use of drugs that are ineffective on individual patients. Material & Methods: Five pancreatic tumour patients operated at Helsinki University Hospital were enrolled in the study. Samples of cancer and normal tissue were taken during the operation. Samples were minced and digested into single cells and cultured into organoids in Matrigel. Pharmacotyping was done with gemcitabine, paclitaxel, FOLFIRINOX and trametinib. DMSO was used as control. We measured vitality after 7 days of incubation. Histopathology showed ductal adenocarcinoma in four patients, one patient had a tubulovillous adenoma with high grade dysplasia. Results: In our experiment, only one patient sample showed sensitivity for gemcitabine (patient 1). The patient in question was sensitive to all drugs tested. In comparison - patients two and three showed a broad drug resistance. Only a high dose of trametinib was able to inhibit growth in these two patient samples. Both gemcitabine and paclitaxel seemed paradoxally to increase growth in organoids derived from patient 2. Patient 4 showed sensitivity to all drugs tested, except for gemcitabine. The patient had received gemcitabine as a neoadjuvant, and no decreasement of tumour size was seen on CT-scan after gemcitabine treatment. Conclusions: Organoids from different patients differed in their responses to drugs. In the case of one patient we could see that gemcitabine resistance in organoids correlated with lack of clinical response to gemcitabine during neoadjuvant treatment. Results were ready 4 weeks after operation and can potentially be considered when adjuvant therapy begins. Relative vitality after drug treatment in organoids from four different patients

Persistence of Human Immunodeficiency Virus-1 Drug Resistance Mutations in Proviral Deoxyribonucleic Acid After Virologic Failure of Efavirenz-Containing Antiretroviral Regimens.

BackgroundEfavirenz (EFV)-based regimens select broad drug resistance to nonnucleoside reverse-transcriptase inhibitors (NNRTIs), limiting the effectiveness of EFV and other NNRTIs. The duration, persistence, and decay of drug resistance mutations (DRMs) in the proviral reservoir is not well defined.MethodsParticipants with virologic failure of EFV-based regimens and drug-resistant viremia with the K103N mutation in plasma ribonucleic acid (RNA) were identified from AIDS Clinical Trials Group (ACTG) studies A364 and A5095. These individuals received a second-line, boosted protease inhibitor-based regimen with suppression of viremia for up to10 years during long-term follow-up (median = 3.6 years; interquartile range, 2.1–6.9 years). Proviral deoxyribonucleic acid (DNA) from cryopreserved peripheral blood mononuclear cells was sequenced to identify the persistence of DRM.ResultsTwenty-eight participants from ACTG 364 and ACTG 5095 were evaluated. Sanger sequencing of proviral DNA detected K103N as well as additional reverse-transcriptase inhibitor (RTI) mutations. Ultradeep sequencing confirmed persistence of K103N in 71% of participants with minimal decay over time. In an adjusted model including years since suppression, persistent proviral K103N was 2.6 times more likely (95% confidence interval, 1.0–6.4) per log10 higher human immunodeficiency virus RNA at EFV failure.ConclusionsPersistence of RTI mutations in proviral DNA after virologic failure has implications for the effectiveness of future drug regimens and the recycling of RTI drugs.

Abstract 1960: T790M-independent EGFR TKI resistance is associated with a broad multi-drug resistant phenotype but selective vulnerabilities to spindle assembly complex (SAC) and CDK inhibitors

Abstract While EGFR mutant NSCLC patients are initially responsive to EGFR targeted therapies, resistant disease inevitably emerges and in nearly half of resistance cases, tumors lack secondary EGFR mutations such as T790M and are refractory to 2nd and 3rd generation EGFR tyrosine kinase inhibitors (TKI). The identification of treatment regimens with efficacy against T790M-negative resistance remains a major clinical challenge. To address this unmet need, we derived a panel of NSCLC cell lines with acquired resistance to the EGFR TKI, erlotinib. A subset of EGFR TKI resistant variants were negative for secondary EGFR mutations, were resistant to 2nd and 3rd generation EGFR TKIs including osimertinb, afatinib, and dacomitinib, and had undergone epithelial to mesenchymal transition (EMT) as demonstrated by loss of E-cadherin, enhanced expression of N-cadherin and Axl and an increased invasive phenotype as determined by Boyden chamber assay. Proteomic profiling revealed that although EGFR TKI resistant cells displayed similar mesenchymal and invasive phenotypes, there was significant heterogeneity in protein expression and pathway activation among resistant variants derived from the same parental cell line. To identify therapeutic agents with activity against EMT-associated EGFR TKI resistance, we performed high-throughput drug screening to test the efficacy of 1,321 compounds. EMT-associated EGFR TKI resistance was accompanied by the acquisition of broad spectrum drug resistance. Compared to parental cells, mesenchymal EGFR TKI resistant cells were significantly more resistant to chemotherapeutic agents used to treat NSCLC including pemetrexed, irinotecan, vinblastine, and gemcitabine. EGFR TKI resistant cells displayed acquired resistance to 147 other tyrosine kinase and serine/threonine kinase inhibitors. In contrast, both parental cells and mesenchymal EGFR TKI resistant variants were highly sensitive to CDK inhibitors and agents targeting spindle assembly checkpoint (SAC) components including PLK1, Aurora, KSP, and survivin. These finding were validated by MTS and clonogenic assays. Treatment with SAC inhibitors induced the accumulation of cells in G2/M phase, enlarged nuclear size, and polyploidy. To clinically validate these findings, we established a cell line (MDA-011) from the pleural effusion of an EGFR mutant NSCLC patient with T790M-negative resistance to erlotinib. In vitro, MDA-011 cells were resistant to erlotinib and osimertinib. MDA-011 cells were highly sensitive to CDK inhibitors and SAC inhibitors as determined by MTS and clonogenic assays. These data indicate that EMT-associated resistance to EGFR TKIs is associated with broad spectrum drug resistance but vulnerabilities to CDK and SAC inhibition which can potentially be exploited to overcome resistant disease in NSCLC patients. Citation Format: Monique B. Nilsson, Jacqueline Robichaux, Huiying Sun, Pan Tong, Lixia Diao, Mia Hofstad, YouHong Fan, Jing Wang, John Heymach. T790M-independent EGFR TKI resistance is associated with a broad multi-drug resistant phenotype but selective vulnerabilities to spindle assembly complex (SAC) and CDK inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1960.