In vitro antibacterial, antibiofilm activities, and phytochemical properties of Posidonia oceanica (L.) Delile: An endemic Mediterranean seagrass.

In the antibiotic resistance era, utilizing understudied sources for novel antimicrobials or antivirulence agents can provide new advances against antimicrobial resistant pathogens. In this study, we aimed to investigate antibacterial and antibiofilm activities of Posidonia oceanica (L.) Delile against Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Klebsiella pneumoniae ATCC 700603 and several S. aureus clinical isolates obtained from medical devices, including patient urinary catheters and breast implant infections, with varying antibiotic recalcitrance profiles. The ethanolic and methanolic extracts from P. oceanica rhizome exhibited significant antibacterial activity against E. faecalis and S. aureus, as well as drug resistant S. aureus clinical isolates. Furthermore, significant antibiofilm activity was observed against S. aureus and E. faecalis treated with ER, MR1, and MR2. P. oceanica extracts also exhibited synergistic antimicrobial activity with ciprofloxacin against E. faecalis, sensitizing E. faecalis to a lower ciprofloxacin concentration. Collectively, our data demonstrate the selective antibacterial and antibiofilm activity of the extracts of P. oceanica against Gram-positive bacteria and clinical isolates along with potentiation of current antibiotics, which suggests that P. oceanica can be further investigated as a potential source for novel therapeutic options in the treatment of drug resistant bacterial infections.

Combination MEG3 lncRNA and Ciprofloxacin dramatically decreases cell migration and viability as well as induces apoptosis in GC cells in vitro.

Gastric cancer (GC) is a prominent cause of cancer-related mortality worldwide. Long noncoding RNA (lncRNA) maternal expression gene3 (MEG3) participates in numerous signaling pathways by targeting the miRNA-mRNA axis. Studies on human tumors have demonstrated that the antibiotic Ciprofloxacin induces cell cycle changes, programmed cell death, and growth suppression. In this study, we transfected MEG3 lncRNA and Ciprofloxacin into the MKN-45 GC cell line. qRT-PCR was employed to evaluate the effects on the specific microRNA and mRNA. The wound healing test, MTT assay, and flow cytometry were used to assess the impact of their administration on cell migration, viability, and apoptosis, respectively. Research showed that miR-147 expression fell even more after MEG3 lncRNA transfection, leading to an increase in B-cell lymphoma 2 (BCL-2) levels. Ciprofloxacin transfection did not significantly affect the axis, except for MEG3, which led to its slight upregulation. MEG3 lncRNA inhibited the migration of MKN-45 cells compared to the control group. When MEG3 lncRNA was coupled with Ciprofloxacin, there was a significant reduction in cell migration compared to untreated groups and controls. MTT assay and flow cytometry demonstrated that MEG3 lncRNA decreased cell viability and triggered apoptosis. Simultaneous administration of MEG3 lncRNA and Ciprofloxacin revealed a significant reduction in cell viability caused by increased apoptosis obtained from MTT or flow cytometry assays. Modulating the miR-147-BCL-2 axis decreases cell migration and survival while promoting cell death. In conclusion, combining MEG3 lncRNA with Ciprofloxacin may be an effective therapeutic approach for GC treatment by influencing the miR-14-BCl-2 axis, resulting in reduced cell viability, migration, and increased apoptosis.

Recurrent Campylobacter jejuni Enteritis in a Patient with Common Variable Immunodeficiency Over an Eight Year Period.

BACKGROUND: In this case, we report multiple isolations of C. jejuni in a patient with common variable immunodeficiency between 2010 and 2018. METHODS: C. jejuni was investigated in the stool samples of the patient by classical culture method using selective media under microaerophilic atmosphere. Antibiotic susceptibilities of the strains were determined by disk diffusion method. RESULTS: Eight C. jejuni strains were isolated from the patient. All strains were resistant to ciprofloxacin. An erythromycin susceptible isolate was replaced by a resistant strain within a one- and four-month period. An erythromycin resistant isolate was replaced by a susceptible one within a year. The patient recovered all episodes by intravenous immunoglobulin replacement and antibiotherapy. CONCLUSIONS: Prolonged or recurrent C. jejuni infections should not be overlooked in immunosuppressed patients. The fact that antibiotic susceptibility may change should also be kept in mind.

Investigating the Significance of Non-jejuni/coli Campylobacter Strains in Patients with Diarrhea.

Campylobacter is one of the most commonly reported foodborne bacteria worldwide. Although Campylobacter jejuni and Campylobacter coli have been reported to be responsible for the great majority of campylobacteriosis, the burden of infections by species other than C. jejuni and C. coli have been increasing as a result of a transition to diagnostic test methods that enable the isolation of emerging species. The aim of the present study was to recover C. jejuni, C. coli, and emerging species from the stool samples of 500 patients with gastroenteritis and 100 healthy subjects via the use of a filtration method and culture techniques using Butzler agar and mCCDA under a microaerobic or hydrogen-enriched atmosphere, identify the species by multiplex PCR methods and assess the significance of emerging species in enteric diseases. Thirty-one (6.2%) Campylobacter spp. were isolated from the stool samples of diarrheic patients but none from healthy individuals. Of 31 isolates, 21 (67.8%), nine (29%), and one (3.2%) were identified as C. jejuni, C. coli, and Campylobacter concisus by multiplex PCR, respectively. The filtration method was superior to the culture technique using mCCDA under a microaerobic atmosphere. C. concisus was evaluated as the etiology of gastroenteritis as a result of laboratory and clinical evaluations. The present study was the first to indicate that emerging Campylobacter species are rarely detected and C. concisus is linked to acute gastroenteritis in Turkey where additional studies are warranted to clarify the significance of emerging species in gastroenteritis.

Prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and Mycoplasma genitalium among Patients with Urogenital Symptoms in Istanbul.

Chlamydia trachomatis, Neisseria gonorrhoeae, and Mycoplasma genitalium are the three most commonly reported sexually transmitted bacteria. The present study aimed to investigate the presence of C. trachomatis, N. gonorrhoeae, and M. genitalium in urogenital samples collected from 18-68-year-old Turkish patients who were admitted to the hospital with various urogenital symptoms. A total of 360 patients with symptoms of STD were included in the study. Following DNA extraction by QIAamp Mini Kit, the presence of C. trachomatis, N. gonorrhoeae, and M. genitalium were investigated using multiplex real-time PCR. Causative organisms were identified in 68 (18.9%) of 360 patients. C. trachomatis, N. gonorrhoeae, and M. genitalium were detected in 40 (11.1%), 14 (3.9%), and 28 (7.8%) of the patients, respectively. Patients 21-30 years of age represented more than one-third (37.8%) of positive patients. Of all patients, dual infections of C. trachomatis-M. genitalium, N. gonorrhoeae-C. trachomatis, N. gonorrhoeae-M. genitalium, and triple infection of C. trachomatis-N. gonorrhoeae-M. genitalium were determined in 1.6% (6/360), 1.3% (5/360), 0.2% (1/360), and 0.2% (1/360) of the patients, respectively. In CT-, NG-, and MG-positive patients, different STI agents were also found such as HIV, HBV, HPV, HSV2, T. pallidum, and T. vaginalis. In conclusion, among C. trachomatis, N. gonorrhoeae, and M. genitalium, CT was the most frequently detected bacterial cause of STDs in our hospital at Istanbul. Co-infections, which comprise more than one-fifth of the cases, should not be underestimated. Regular screening and following up of STD agents using multiplex real-time PCR-based diagnostic methods enabling the immediate detection of co-infections are essential for the treatment and primary prevention of STDs.

Interplay between HIV and Human Pegivirus (HPgV) Load in Co-Infected Patients: Insights from Prevalence and Genotype Analysis.

Human pegivirus (HPgV) is transmitted through sexual or parenteral exposure and is common among patients receiving blood products. HPgV is associated with lower levels of human immunodeficiency virus (HIV) RNA and better survival among HIV-infected patients. This study aimed to investigate the prevalence of HPgV and determine its subtypes in HIV-infected individuals living in Istanbul, which has the highest rate of HIV infection in Türkiye. Total RNA extraction from plasma, cDNA synthesis, and nested PCR were performed for HPgV on plasma samples taken from 351 HIV-1-infected patients. The HPgV viral load was quantified on HPgV-positive samples. HPgV genotyping was performed by sequencing the corresponding amplicons. In the present study, the overall prevalence of HPgV RNA in HIV-infected patients was 27.3%. HPgV subtypes 1, 2a, and 2b were found, with subtype 2a being the most frequent (91.6%). Statistical analysis of HIV-1 viral load on HPgV viral load showed an opposing correlation between HIV-1 and HPgV loads. In conclusion, these data show that HPgV infection is common among HIV-positive individuals in Istanbul, Türkiye. Further comprehensive studies are needed to clarify both the cellular and molecular pathways of these two infections and to provide more information on the effect of HPgV on the course of the disease in HIV-infected individuals.

A public awareness study about antibiotic consumption habits and antibacterial resistance in north-eastern region of Cyprus.

INTRODUCTION: Antibiotic resistance is a current global issue. Investigation of the level of knowledge of the public about antibiotics and antibiotic resistance is necessary to combat the antibiotic resistance problem. The aim of the present study was to evaluate the level of knowledge of the citizens of the north-eastern part of Cyprus on antibiotics and antibiotic resistance problem. METHODOLOGY: Randomly selected 701 adults were included in the study. A modified version of World Health Organization's public awareness survey was used to assess the knowledge on antibiotics and the resistance. Logistic regression was used to find out the relationship between knowledge and education level. Spearman's correlation analysis was carried out to determine the association between the education level and the awareness of antibiotic resistance. RESULTS: Overall, 47.9% (336/701) of the respondents had used antibiotics in the last 6 months. Approximately 70% of respondents were determined to have intermediate/high knowledge on antibiotic consumption. In total, 66% of the population heard about antibiotic resistance and of these, 64% had intermediate knowledge on the resistance concept. University graduates were more likely to hear the term antibiotic resistance than primary school graduates. CONCLUSIONS: In the north-eastern region of Cyprus, the public is moderately knowledgeable about antibiotics and resistance. The study is the first large scale study in the northern part of Cyprus and is thought to improve the national health policies related with antibiotic consumption in Cyprus and other developing countries.

Molecular Typing of Campylobacter jejuni and Campylobacter coli of Human Strains Isolated in Turkey Over an Eight-Year Period.

BACKGROUND: Campylobacter spp. is one of the leading causes of bacterial foodborne infections worldwide. In this study, we aimed to investigate the genetic diversity of 341 Campylobacter strains isolated in Turkey. METHODS: Campylobacter spp. was identified by phenotypical methods and PCR. Species level identification was carried out by the hippurate hydrolysis test and PCR. C. jejuni and C. coli strains were typed by using flaA-RFLP and PFGE. RESULTS: Of 341 strains, 300 (88%), 37 (10.8%), and four were identified as C. jejuni, C. coli, and non-jejuni/non-coli, respectively. The hippurate hydrolysis test misidentified 12% of 341 strains. The typeabilities of flaA-RFLP and PFGE were 100% for C. coli, whereas those of flaA-RFLP and PFGE for C. jejuni were 99.3% and 99%, respectively. The discriminatory power of the combination of PFGE and flaA-RFLP was determined to be higher than either method alone for both C. jejuni and C. coli. Both of the strains were so diverse that 80% and 64% of C. jejuni and C. coli genotypes included only one strain, respectively. In two patients, Campylobacter strains that were isolated from the first stool samples were C. jejuni where as those isolated from the second samples, collected eight and 20 days after the collection of the first samples, were C. coli. C. jejuni strains that were recovered from two different stool samples of two patients, collected 1 - 2 days apart, were found to be genetically different. CONCLUSIONS: Species identification of Campylobacter strains should be done using molecular methods. Combination of two methods is prerequisite for increasing the accuracy of molecular typing. Mixed or subsequent infection by different Campylobacter species and C. jejuni of different genotypes should not be underestimated.

Molecular and phenotypical investigation of ciprofloxacin resistance among Campylobacter jejuni strains of human origin: high prevalence of resistance in Turkey.

Campylobacteriosis is one of the most frequently reported zoonoses worldwide. The well-documented increase in the ciprofloxacin resistance has increased the importance of rapid detection of the resistance. The incidence of ciprofloxacin resistance was investigated using real-time PCR. Identification of one hundred and fifty-eight strains was performed by PCR. Minimum inhibitory concentration (MIC) of ciprofloxacin was determined by Epsilometer test. Following the confirmation of the efficiencies of singleplex real-time PCR methods using two different probes, a cytosine to thymine point mutation at codon 86 was detected by allelic discrimination. Of the 158 strains, 114 (72.2%) were determined to be resistant to ciprofloxacin. The MIC50 and the MIC90 of ciprofloxacin were found to be 8 and ≥32 mg/L, respectively. By real-time PCR, the presence of the mutation was confirmed in all, but one, resistant strains and the absence of the mutation was demonstrated in all, but one, susceptible strains. The rate of resistance is high among C. jejuni strains and ciprofloxacin should not be used in the treatment of such infections in Turkey. A cytosine to thymine mutation is the most frequently detected mechanism for the resistance. Real-time PCR can be used for the quick screening of the resistance.

The fate of suboptimal anastomosis after colon resection: An experimental study.

BACKGROUND: The fate of suboptimal anastomosis is unknown and early detection of anastomotic leakage after colon resection is crucial for the proper management of patients. METHODS: Twenty-six rats were assigned to "Control", "Leakage" and "Suboptimal anastomosis" groups where they underwent either sham laparotomy, cecal ligation, and puncture or anastomosis with four sutures following colon resection, respectively. At the fifth hour and on the third and ninth days; peripheral blood and peritoneal washing samples through relaparotomy were obtained. The abdomen was inspected macroscopically for anastomotic healing. Polymerase chain reaction (PCR) with 16s rRNA and E.coli-specific primers were run on all samples along with aerobic and anaerobic cultures. RESULTS: The sensitivity and specificity of PCR on different bodily fluids with 16s rRNA and E.coli-specific primers were 100% and 78%, respectively. All samples of peritoneal washing fluids on the third and ninth days showed presence of bacteria in both PCR and culture. The inspection of the abdomen revealed signs of anastomotic leakage in eight rats (80%), whereas mortality related with anastomosis was detected in two (20%). CONCLUSION: Anastomotic leakage with suboptimal anastomosis after colon resection is high and the early detection is possible by running PCR on peritoneal samples as early as 72 hours.

Carbapenem resistance due to Bla(OXA-48) among ESBL-producing Escherichia coli and Klebsiella pneumoniae isolates in a univesity hospital, Turkey.

Bacterial isolates producing Class D OXA-48 carbapenemase may be missed in routine laboratory testing, allowing them to spread undetected. The purpose of the present study was to detect bla(OXA-48) among ESBL-producing Klebsiella pneumoniae and Escherichia coli isolates collected from a university hospital, Turkey. Ninety-two ESBL-producing isolates (66 E. coli, 26 K. pneumoniae) were obtained in 2010. Antibiotic susceptibility tests were performed using the disc diffusion method and VITEK 2 system. Carbapenemase activity was screened using modified Hodge test. Beta-lactamase genes were detected by PCR and bla(OXA-48)-positive amplicons were sequenced. Genetic relatedness among K. pneumoniae isolates was investigated by pulsed-field gel-electrophoresis (PFGE). Carbapenemase activity was detected in 1 E. coli and 9 K. pneumoniae isolates and 8 of the K. pneumoniae plus the E. coli isolates were resistant to ertapenem. Three K. pneumoniae and 1 E. coli isolates were resistant to imipenem. All 10 isolates were susceptible to meropenem. bla(OXA-48) was present in all 10 isolates. Additionally, 9 isolates contained at least one beta-lactamase gene, including bla(SHV') bla(CTX-M) and bla(VEB) type. PFGE revealed different karyotypes among 9 K. pneumoniae isolates suggesting that the dissemination of bla(OXA-48) gene was not spread by a single K. pneumoniae clone. Thus OXA-48-producing isolates found in carbapenem-susceptible strains according to CLSI guidelines.