COMBINATORIAL EFFECT OF D-AMINOACIDS AND TETRACYCLINE AGAINST PSEUDOMONAS AERUGINOSA BIOFILM

Objective: The present study attempted to evaluate the anti-biofilm activity of D-amino acids (D-AAs) on Pseudomonas aeruginosa and determine if the combination of D-AAs with tetracycline enhances the anti-biofilm activity in vitro and ex vivo.Methods: Different D-AAs were tested for antibiofilm activity against wild type P. aeruginosa PAO1 and two multidrug resistant P. aeruginosa clinical strains in the presence of sub inhibitory concentrations of tetracycline using crystal violet microtitre plate assay. Results were further validated using in vitro wound dressing and ex vivo porcine skin models followed by cytotoxicity and hemocompatibility studies.Results: D-tryptophan (5 mmol) showed 61 % reduction in biofilm formation of P. aeruginosa. Interestingly combinatorial effect of 5 mmol D-tryptophan and 0.5 minimum inhibitory concentration (MIC) (7.5µg/ml) tetracycline showed 90% reduction in biofilm formation. 5 mmol D-methionine shows 28 % reduction and combination with tetracycline shows 41% reduction in biofilm formation of P. aeruginosa. D-leucine and D-tyrosine alone or in combination with tetracycline did not show significant anti-biofilm activity. D tryptophan-tetracycline combination could reduce 80 % and 77 % reduction in biofilm formation in two multi drug resistant P. aeruginosa clinical strains. D-tryptophan-tetracycline-combination could also reduce 76% and 66% reduction in biofilm formation in wound dressing model and porcine skin explant respectively. The cytotoxicity and hemocompatibility studies did not show significant toxicity when this combination was used.Conclusion: The results established the potential therapeutic application of D-tryptophan alone or in combination with tetracycline for treating biofilm associated clinical problems caused by P. aeruginosa

Вирусные гепатиты В, С и D в Республике Молдова: достижения и проблемы

Agenția Națională pentru Sănătate Publică, IP Universitatea de Stat de Medicină și Farmacie Nicolae Testemițanu, IMSP Centrul Republican de Diagnosticare MedicalăRealizarea primelor trei Programe Naționale de combatere a hepatitelor virale B, C și D în Republica Moldova, începând cu anul 1997, au condus la reducerea semnificativă a morbidității prin hepatitele virale B, C și D acute, dar pe fundalul incidenței sporite a hepatitelor virale cronice, unde indicatorii îi depășesc vădit pe cei din țările Comunității Europene. Studiile realizate demonstrează că contingentele de populație cu risc sporit de infectare continuă să prezinte un potențial înalt ca sursă reală de infectare cu virusurile hepatice nominalizate pentru alte categorii de populație. Ca urmare a utilizării tehnicilor de biologie moleculară, au fost evidențiate genotipurile și subgenotipurile virusurilor hepatitelor B, C și D în special la contingentele de populație cu risc sporit de infectare, demonstrând că unele posedă un potențial înalt de cronicizare, cu evoluție spre ciroză și cancer hepatocelular. Utilizarea produselor antivirale de ultimă generație recomandate de OMS pentru tratamentul hepatitelor virale B și C a demonstrat o eficacitate înaltă nu numai terapeutică, ci și profi lactică, manifestată prin reducerea numărului de bolnavi purtători de virusuri și a noilor focare. Este important de menționat că prejudiciul economic prevenit ca urmare a realizării Programelor Naționale de combatere a hepatitelor virale în perioada 1997-2015 a constituit 822.453 mii lei, pe când cheltuielile din bugetul de stat pentru realizarea acestor programe au alcătuit 84.618,5 mii lei. Realizarea integrală a prezentului Program de combatere a hepatitelor virale B, C și D în contextul Planului Strategic al OMS de eliminare a hepatitelor virale parenterale până în anul 2030 va avea un beneficiu socioeconomic extrem de important pentru Republica Moldova.The implementation of the first three National Programs against viral hepatitis B, C and D in the Republic of Moldova since 1997 has allowed to significantly reduce morbidity of acute viral hepatitis B, C and D, but on the increased background of incidence of chronic viral hepatitis the indicators clearly outweigh those in the European Community. Studies have shown that the contingent of population at high risk of infection continues to show a high potential as a real source of infection with viral hepatitis viruses for other population categories. As a result of the use of molecular biology techniques, the genotypes and subgenotypes of hepatitis B, C and D viruses have been highlighted, particularly in contingent of population at high risk of infection, demonstrating that some have a high potential for chronicization with progression to cirrhosis and hepatocellular cancer. Using of next-generation antiviral drugs in the treatment of viral hepatitis B and C recommended by the WHO has shown a high, not only therapeutic but also prophylactic efficacy manifested by reducing the number of virus-bearing patients, and new outbreaks. It is important to note that the economic prejudice prevented by the implementation of the National Programs against viral hepatitis during the period 1997-2015 amounted to 822.453 thousand lei, while the spending from the State Budget for the implementation of the nominated programs amounted to 84.618.5 thousand lei. The full implementation of the current Program for combating viral hepatitis B, C and D in the context of the WHO Strategic Plan for the elimination of parenteral viral hepatitis by 2030 will have a very important socio-economic benefit for the Republic of Moldova.Реализация первых трех Национальных программ по борьбе с вирусными гепатитами B, C и D в Республике Молдова с 1997 года позволила значительно снизить заболеваемость острыми вирусными гепатитами B, C и D, но на фоне высокого уровня заболеваемости хроническими вирусными гепатитами, показатели явно перевешивают показатели стран Европейского Cоюза. Исследования показали, что среди групп населения с высоким риском инфицирования все еще присутствует высокий потенциал в качестве реального источника заражения вирусами гепатитов для других категорий населения. В результате использования методов молекулярной биологии были выделены генотипы и субгенотипы вирусов гепатитов B, C и D, особенно среди групп с высоким риском инфицирования, демонстрируя, что некоторые имеют высокий потенциал к хронизации и прогрессированию к циррозу и гепатоцеллюлярного рака. Использование противовирусных препаратов последнего поколения для лечения вирусных гепатитов В, C и D, рекомендованных ВОЗ, продемонстрировало не только высокую терапевтическую, но и профилактическую эффективность за счет сокращения числа вирусоносителей и новых очагов инфекции. Важно отметить, что предотвращённый экономический ущерб в результате реализации Национальных программ по борьбе с вирусными гепатитами в период 1997–2015 годов составил 822.453 тысяч леев, а расходы государственного бюд-жета на реализацию указанных программ составили 84.618,5 тысяч леев. Полная реализация текущей Программы по борьбе с вирусными гепатитами B, C и D в контексте Стратегического плана ВОЗ по элиминации парентеральных вирусных гепатитов к 2030 году будет иметь важное социально-экономическое значение для Республики Молдова

Clinical and molecular practice of European thoracic pathology laboratories during the COVID-19 pandemic. The past and the near future.

This study evaluated the consequences in Europe of the COVID-19 outbreak on pathology laboratories orientated toward the diagnosis of thoracic diseases. A survey was sent to 71 pathology laboratories from 21 European countries. The questionnaire requested information concerning the organization of biosafety, the clinical and molecular pathology, the biobanking, the workload, the associated research into COVID-19, and the organization of education and training during the COVID-19 crisis, from 15 March to 31 May 2020, compared with the same period in 2019. Questionnaires were returned from 53/71 (75%) laboratories from 18 European countries. The biosafety procedures were heterogeneous. The workload in clinical and molecular pathology decreased dramatically by 31% (range, 3%-55%) and 26% (range, 7%-62%), respectively. According to the professional category, between 28% and 41% of the staff members were not present in the laboratories but did teleworking. A total of 70% of the laboratories developed virtual meetings for the training of residents and junior pathologists. During the period of study, none of the staff members with confirmed COVID-19 became infected as a result of handling samples. The COVID-19 pandemic has had a strong impact on most of the European pathology laboratories included in this study. Urgent implementation of several changes to the organization of most of these laboratories, notably to better harmonize biosafety procedures, was noted at the onset of the pandemic and maintained in the event of a new wave of infection occurring in Europe

Clinical and molecular practice of European thoracic pathology laboratories during the COVID-19 pandemic The past and the near future

BackgroundThis study evaluated the consequences in Europe of the COVID-19 outbreak on pathology laboratories orientated toward the diagnosis of thoracic diseases.Materials and methodsA survey was sent to 71 pathology laboratories from 21 European countries. The questionnaire requested information concerning the organization of biosafety, the clinical and molecular pathology, the biobanking, the workload, the associated research into COVID-19, and the organization of education and training during the COVID-19 crisis, from 15 March to 31 May 2020, compared with the same period in 2019.ResultsQuestionnaires were returned from 53/71 (75%) laboratories from 18 European countries. The biosafety procedures were heterogeneous. The workload in clinical and molecular pathology decreased dramatically by 31% (range, 3%-55%) and 26% (range, 7%-62%), respectively. According to the professional category, between 28% and 41% of the staff members were not present in the laboratories but did teleworking. A total of 70% of the laboratories developed virtual meetings for the training of residents and junior pathologists. During the period of study, none of the staff members with confirmed COVID-19 became infected as a result of handling samples.ConclusionsThe COVID-19 pandemic has had a strong impact on most of the European pathology laboratories included in this study. Urgent implementation of several changes to the organization of most of these laboratories, notably to better harmonize biosafety procedures, was noted at the onset of the pandemic and maintained in the event of a new wave of infection occurring in Europe

Mechanism and functional consequences of loss of FOXO1 expression in endometrioid endometrial cancer cells

The forkhead transcription factor FOXO1, a downstream target of phosphatidylinositol-3-kinase/Akt signalling pathway, regulates cyclic differentiation and apoptosis in normal endometrium, but its role in endometrial carcinogenesis is unknown. Screening of endometrial cancer cell lines demonstrated that FOXO1 is expressed in HEC-1B cells, but not in Ishikawa cells, which in turn highly express the FOXO1 targeting E3-ubiquitin ligase Skp2. FOXO1 transcript levels were also lower in Ishikawa cells and treatment with the proteasomal inhibitor was insufficient to restore expression. Lack of FOXO1 expression in Ishikawa cells was not accounted for by differential promoter methylation or activity, but correlated with increased messenger RNA (mRNA) turnover. Comparative analysis demonstrated that HEC-1B cells proliferate slower, but are more resistant to paclitaxel-mediated cell death than Ishikawa cells, which were partially reversed upon silencing of FOXO1 in HEC-1B cells or its re-expression in Ishikawa cells. We further show that FOXO1 is required for the expression of the growth arrest- and DNA-damage-inducible gene GADD45α. Analysis of biopsy samples demonstrated a marked loss of FOXO1 and GADD45α mRNA and protein expression in endometrioid endometrial cancer compared to normal endometrium. Together, these observations suggest that loss of FOXO1 perturbs endometrial homeostasis, promotes uncontrolled cell proliferation and increases susceptibility to genotoxic insults