Erythrocyte-Leveraged Oncolytic Virotherapy (ELeOVt): Oncolytic Virus Assembly on Erythrocyte Surface to Combat Pulmonary Metastasis and Alleviate Side Effects.

Despite being a new promising tool for cancer therapy, intravenous delivery of oncolytic viruses (OVs) is greatly limited by poor tumor targeting, rapid clearance in the blood, severe organ toxicity, and cytokine release syndrome. Herein, a simple and efficient strategy of erythrocyte-leveraged oncolytic virotherapy (ELeOVt) is reported, which for the first time assembled OVs on the surface of erythrocytes with up to near 100% efficiency and allowed targeted delivery of OVs to the lung after intravenous injection to achieve excellent treatment of pulmonary metastases while greatly improving the biocompatibility of OVs as a drug. Polyethyleneimine (PEI) as a bridge to assemble OVs on erythrocytes also played an important role in promoting the transfection of OVs. It is found that ELeOVt approach significantly prolonged the circulation time of OVs and increased the OVs distribution in the lung by more than tenfold, thereby significantly improving the treatment of lung metastases while reducing organ and systemic toxicity. Taken together, these findings suggest that the ELeOVt provides a biocompatible, efficient, and widely available approach to empower OVs to combat lung metastasis.

Fimbria targeting superparamagnetic iron oxide nanoparticles enhance the antimicrobial and antibiofilm activity of ciprofloxacin against quinolone-resistant E. coli.

High quinolone resistance of Escherichia coli limits the therapy options for urinary tract infection (UTI). In response to the urgent need for efficient treatment of multidrug-resistant infections, we designed a fimbriae targeting superparamagnetic iron oxide nanoparticle (SPION) delivering ciprofloxacin to ciprofloxacin-resistant E. coli. Bovine serum albumin (BSA) conjugated poly(acrylic acid) (PAA) coated SPIONs (BSA@PAA@SPION) were developed for encapsulation of ciprofloxacin and the nanoparticles were tagged with 4-aminophenyl-α-D-mannopyrannoside (mannoside, Man) to target E. coli fimbriae. Ciprofloxacin-loaded mannoside tagged nanoparticles (Cip-Man-BSA@PAA@SPION) provided high antibacterial activity (97.1 and 97.5%, respectively) with a dose of 32 µg/mL ciprofloxacin against two ciprofloxacin-resistant E. coli isolates. Furthermore, a strong biofilm inhibition (86.9% and 98.5%, respectively) was achieved in the isolates at a dose 16 and 8 times lower than the minimum biofilm eradication concentration (MBEC) of ciprofloxacin. Weaker growth inhibition was observed with untargeted nanoparticles, Cip-BSA@PAA@SPIONs, confirming that targeting E. coli fimbria with mannoside-tagged nanoparticles increases the ciprofloxacin efficiency to treat ciprofloxacin-resistant E. coli. Enhanced killing activity against ciprofloxacin-resistant E. coli planktonic cells and strong growth inhibition of their biofilms suggest that Cip-Man-BSA@PAA@SPION system might be an alternative and/or complementary therapeutic option for the treatment of quinolone-resistant E. coli infections.

BNT162b2 or CoronaVac as the Third Dose against Omicron: Neutralizing Antibody Responses among Transplant Recipients Who Had Received Two Doses of CoronaVac.

We evaluated neutralizing antibodies against the Omicron variant and Anti-Spike IgG response in solid organ (SOT) or hematopoietic stem cell (HSTC) recipients after a third dose of BNT162b2 (BNT) or CoronaVac (CV) following two doses of CV. In total, 95 participants underwent SOT (n = 62; 44 liver, 18 kidney) or HSCT (n = 27; 5 allogeneic, 22 autologous) were included from five centers in Turkey. The median time between third doses and serum sampling was 154 days (range between 15 to 381). The vaccine-induced antibody responses of both neutralizing antibodies and Anti-Spike IgGs were assessed by plaque neutralizing assay and immunoassay, respectively. Neutralizing antibody and Anti-Spike IgG levels were significantly higher in transplant patients receiving BNT compared to those receiving CV (Geometric mean (GMT):26.76 vs. 10.89; p = 0.03 and 2116 Au/mL vs. 172.1 Au/mL; p < 0.001). Solid organ transplantation recipients, particularly liver transplant recipients, showed lower antibody levels than HSCT recipients. Thus, among HSCT recipients, the GMT after BNT was 91.29 and it was 15.81 in the SOT group (p < 0.001). In SOT, antibody levels after BNT in kidney transplantation recipients were significantly higher than those in liver transplantation recipients (GMT: 48.32 vs. 11.72) (p < 0.001). Moreover, the neutralizing antibody levels after CV were very low (GMT: 10.81) in kidney transplantation recipients and below the detection limit (<10) in liver transplant recipients. This study highlights the superiority of BNT responses against Omicron as a third dose among transplant recipients after two doses of CV. The lack of neutralizing antibodies against Omicron after CV in liver transplant recipients should be taken into consideration, particularly in countries where inactivated vaccines are available in addition to mRNA vaccines.

Virus-Protein Corona Replacement Strategy to Improve the Antitumor Efficacy of Intravenously Injected Oncolytic Adenovirus.

Intravenous administration of oncolytic adenoviruses (OVs) is a hopeful tumor therapeutic modality. However, the sharp clearance of OVs by the immune system dampens its effectiveness. Many studies have attempted to extend the circulation of intravenously administered OVs, almost all by preventing OVs from binding to neutralizing antibodies and complements in the blood, but the results have not been satisfactory. In contrast to previous conclusions, we found that the key to improving the circulation of OVs is to prevent the formation of the virus-protein corona rather than simply preventing the binding of neutralizing antibodies or complements to OVs. After identifying the key protein components of the virus-protein corona, we proposed a virus-protein corona replacement strategy, where an artificial virus-protein corona was formed on OVs to completely prevent the interaction of OVs with key virus-protein corona components in the plasma. It was found that this strategy dramatically prolonged the circulation time of OVs by over 30 fold and increased the distribution of OVs in tumors by over 10-fold, resulting in superior antitumor efficacy in primary and metastatic tumor models. Our finding provides a perspective on intravenous delivery of OVs, shifting the focus of future studies from preventing OV binding with neutralization antibodies and complements to preventing OVs from interacting with key virus-protein corona components in the plasma.

The rapid diagnosis of viral respiratory tract infections and its impact on antimicrobial stewardship programs.

We aimed to describe the potential benefit of new rapid molecular respiratory tests (MRT) in decreasing inappropriate antibiotic use among the inpatients presenting with influenza-like illness (ILI). We included patients from inpatient and outpatient departments who had ILI and performed MRT between 1 January 2015 and 31 December 2016 in a 265-bed private hospital in Istanbul. At the end of 2015, we implemented antimicrobial stewardship including systematic use of MRT. Then, we compared our observations between the year 2015 and the year 2016. We designed the study according to the STrengthening the Reporting of OBservational studies in Epidemiology (STROBE) tool. A U.S. Food and Drug Administration (FDA)-cleared multiplexed polymerase chain reaction (PCR) system (BioFire FilmArray, Idaho Technology, Salt Lake City, UT) which detects 17 viruses and three bacteria was used for diagnosis. In total, 1317 patients were included; 630 (48%) were inpatients and 569 (43%) were older than 16 years of age. At least one virus was detected in 747 (57%) patients. Rhinovirus/enterovirus, influenza virus, and adenovirus were the most commonly detected. Among hospitalized patients, in children, a significant decrease in antibiotic use (44.5% in 2015 and 28.8% in 2016, p = 0.009) was observed, but in adults, the decrease was not statistically significant (72% in 2015 and 63% in 2016, p = 0.36). The duration of antibiotic use after the detection of virus was significantly decreased in both children and adults (p < 0.001 and p = 0.007, respectively). By using MRT, inappropriate antibiotic use and, also, duration of inappropriate antibiotic use after the detection of virus was significantly decreased. It is time to increase the awareness about the viral etiology in respiratory tract infections (RTIs) and implement MRT in clinical practice.

Cytokine response in crimean-congo hemorrhagic fever virus infection.

We described the predictive role of cytokines in fatality of Crimean Congo Hemorrhagic Fever Virus (CCHFV) infection by using daily clinical sera samples. Consequent serum samples of the selected patients in different severity groups and healthy controls were examined by using human cytokine 17-plex assay. We included 12 (23%) mild, 30 (58%) moderate, 10 (19%) severe patients, and 10 healthy volunteers. The mean age of the patients was 52 (sd 15), 52% were female. Forty-six patients (88%) received ribavirin. During disease course, the median levels of IL-6, IL-8, IL-10, IL-10/12, IFN-γ, MCP-1, and MIP-1b were found to be significantly higher among CCHF patients than the healthy controls. Within the first 5 days after onset of disease, among the fatal cases, the median levels of IL-6 and IL-8 were found to be significantly higher than the survived ones (Fig. 3), and MCP-1 was elevated among fatal cases, but statistical significance was not detected. In receiver operating characteristic (ROC) analysis, IL-8 (92%), IL-6 (92%), MCP-1 (79%) were found to be the most significant cytokines in predicting the fatality rates in the early period of the disease (5 days). IL-6 and IL-8 can predict the poor outcome, within the first 5 days of disease course. Elevated IL-6 and IL-8 levels within first 5 days could be used as prognostic markers.

The clinical impact of ST131 H30-Rx subclone in urinary tract infections due to multidrug-resistant Escherichia coli.

In this study, risk factors for ST131 H30 and H30-Rx subclones among urinary tract infections (UTIs) caused by multidrug-resistant (MDR) Escherichia coli were described. Urine samples were collected from consecutive outpatients registered to the outpatient clinics of Baskent University Hospital (Ankara, Turkey) with complaints of acute cystitis in 2011. A total of 107 MDR E. coli isolates were included in the study. Of the 107 isolates studied, 26 (24.3%) were typed as ST131 clone. Extended-spectrum ß-lactamase (ESBL)-producers accounted for 59 (55.1%) of the 107 isolates. Among the 59 ESBL-positive isolates, 18 (31%) were found to belong to the ST131 clone. Of the 18 ESBL-positive ST131 isolates, 17 (94%) were defined as H30 subclone, among which 16 (94%) represented the H30-Rx subclone. Among the 48 ESBL-negative isolates, 8 (17%) ST131 isolates were detected, 7 (88%) of which belonged to H30 subclone; 5 (71%) of the H30 subclone isolates were classified under H30-Rx subclone. In multivariate analysis, hospitalisation within last year was the only host risk factor associated with MDR E. coli ST131 H30-Rx subclone UTI (OR=3.5, 95% CI 1.04-12.17; P=0.042). CTX-M-15 production was found to be highly associated with the presence of ST131 H30-Rx subclone (OR=4.8, 95% CI 1.54-15.32; P=0.007). In conclusion, urinary MDR E. coli ST131 H30-Rx subclone was found to be important in the dissemination of MDR UTIs in the community. Approximately 20% of the MDR isolates were H30-Rx subclone. Infection with this subclone was found to be healthcare-associated.

Emerging Escherichia coli O25b/ST131 clone predicts treatment failure in urinary tract infections.

BACKGROUND: We described the clinical predictive role of emerging Escherichia coli O25b/sequence type 131 (ST131) in treatment failure of urinary tract infection. METHODS: In this prospective observational cohort study, the outpatients with acute cystitis with isolation of E. coli in their urine cultures were assessed. All the patients were followed up for clinical cure after 10 days of treatment. Detection of the E. coli O25:H4/ST131 clone was performed by multiplex polymerase chain reaction (PCR) for phylogroup typing and using PCR with primers for O25b rfb and allele 3 of the pabB gene. RESULTS: In a cohort of patients with diagnosis of acute urinary cystitis, 294 patients whose urine cultures were positive with a growth of >10(4) colony-forming units/mL of E. coli were included in the study. In empiric therapy, ciprofloxacin was the first choice of drug (27%), followed by phosphomycin (23%), trimethoprim-sulfamethoxazole (TMP-SMX) (9%), and cefuroxime (7%). The resistance rate was 39% against ciprofloxacin, 44% against TMP-SMX, and 25% against cefuroxime. Thirty-five of 294 (12%) isolates were typed under the O25/ST131 clone. The clinical cure rate was 85% after the treatment. In multivariate analysis, detection of the O25/ST131 clone (odds ratio [OR], 4; 95% confidence interval [CI], 1.51-10.93; P = .005) and diabetes mellitus (OR, 2.1; 95% CI, .99-4.79; P = .05) were found to be significant risk factors for the treatment failure. In another multivariate analysis performed among quinolone-resistant isolates, treatment failure was 3 times more common among the patients who were infected with ST131 E. coli (OR, 3; 95% CI, 1.27-7.4; P = .012). CONCLUSIONS: In urinary tract infections, the E. coli ST131 clone seems to be a consistent predictor of treatment failure.

Risk factors for candidemia with non-albicans Candida spp. in intensive care unit patients with end-stage renal disease on chronic hemodialysis.

BACKGROUND/PURPOSE: The objective of this study was to describe factors associated with bloodstream infections (BSIs) with non-albicans Candida species (NAC), compared with Candida albicans BSIs, and antifungal susceptibility patterns in adult intensive care unit (ICU) patients with chronic renal failure undergoing hemodialysis. To the best of our knowledge, this is the first study to report the potential factors for NAC candidemia in ICU patients with end-stage renal disease on chronic hemodialysis. METHODS: This prospective, observational, multicenter study was conducted in the two centers of Baskent University between January 2007 and July 2010. All adult patients excluding patients with neutropenia, malignancy, glucocorticoid treatment or AIDS, were included. RESULTS: Sixty cases (58.8%) of candidemia were due to C. albicans and 42 (41.2%) to NAC. Multivariate regression analysis revealed that the presence of a central venous catheter was the only risk factor independently associated with BSI due to NAC (p=0.046, odds ratio: 5.90, 95% confidence interval: 1.032-33.717). Mortality was more frequent in those with NAC than C. albicans BSIs (64.3% vs. 55%), but the difference was not significant (p=0.067). Except for two Candida glabrata strains, which were dose-dependently fluconazole susceptible, all Candida species were susceptible to fluconazole, caspofungin, voriconazole and amphotericin B. CONCLUSION: Central venous catheterization was the only factor significantly associated with BSI due to NAC in ICU patients with end-stage renal disease.

Lymph node human papillomavirus DNA positivity in uterine cervical cancers and its relationship with prognostic factors.

INTRODUCTION: Uterine cervical cancer is a major health care issue worldwide. Human papillomavirus (HPV) DNA positivity in regional lymph nodes has been proposed as a risk factor for recurrence in a number of studies. The aim of our study was to determine the correlation between well-known prognostic factors and HPV DNA positivity in regional lymph nodes of patients with cervical cancer who were treated surgically. METHODS: Thirty-seven patients who underwent radical hysterectomy (Piver class III) with regional lymph node dissection for International Federation of Gynecology and Obstetrics stage IB uterine cervical cancer were assessed for nodal HPV DNA status using polymerase chain reaction. Human papillomavirus DNA presence and types were analyzed in paraffin-embedded tissues from all primary tumors and 746 regional lymph nodes. The correlation between HPV DNA positivity of the lymph nodes and prognostic factors (stage at the time of diagnosis, status of the regional nodes, status of parametrial and surgical margins, tumor size, histological type, cervical stromal invasion depth, and lymphovascular space invasion was investigated. RESULTS: Lymph node HPV DNA positivity increased in larger tumors (P < 0.05). In addition, lymph node metastasis and primary tumor HPV DNA status were closely correlated with the HPV DNA status of the lymph nodes (P < 0.05). CONCLUSIONS: Human papillomavirus DNA presence in the lymph nodes of patients with uterine cervical cancer may have an influence on disease survival and requires further research. Future studies with prospective design and large sample sizes using fresh operative specimens and quantitative HPV DNA detection methods are needed.

Effect of manual bowel decompression (milking) in the obstructed small bowel.

BACKGROUND: Mechanical intestinal obstruction is a frequently encountered problem in general surgery. One of the frequently used techniques for surgical decompression, so-called milking, is to caress the intestinal contents cephalad into the stomach or caudally into the colon. The aim of our study was to examine the functional, morphologic, and microbiologic effects of manual bowel decompression (milking) in the obstructed small bowel. METHODS: Six rats in the milking (M) group (obstructed and decompressed manually), 6 in the control (C) group (obstructed only), and 5 rats in the sham (S) group (laparotomy and evisceration) underwent surgery. Muscle contractility, gastrointestinal dye transmission, histopathologic changes of ileum, and bacterial translocation were analyzed. RESULTS: The contractile response of intestinal segments to acetylcholine was significantly less in group M compared with those of groups C and S (P < .05). The maximal contractile response to acetylcholine also was significantly lower in group M (P < .05). There was no statistically significant difference between the groups regarding the sensitivity of cholinergic receptors. Frequency of peristaltic movements, progression of Evans blue dye, histopathologic variables, and the quantitative evaluation (colony-forming units/gram of tissue) of isolated bacteria were not different among the groups. However, Escherichia coli in group M and Klebsiella spp in group S were the main isolated bacteria. CONCLUSIONS: Although it reduces muscle contractility, a milking procedure in an intestinal obstruction model does not cause peristaltic deterioration, histopathologic or inflammatory changes, or alterations in the degree of bacterial translocation.

Clinical subgroups and antifungal susceptibilities in fungal culture-positive patients with chronic rhinosinusitis.

The goal of this study was to investigate clinical characteristics of chronic rhinosinusitis (CRS) in patients with positive fungal cultures. Fungal cultures were taken from 55 CRS patients and 20 healthy volunteers. Susceptibilities of isolated fungi to fluconazole, amphotericin B, itraconazole, voriconazole, and caspofungin were determined in CRS patients. Fungi grew in the cultures from 44 (80%) CRS patients and 17 (85%) healthy volunteers. Of the patients with positive fungal cultures, 5 (11.3%) had fungal hypersensitivity (FH), and 21 (47.7%) had eosinophilic mucin (EM). Fungal culture-positive patients with EM were more likely to be associated with presence of polyps and higher CT scores than those without EM (P < 0.05). All the patients with FH had EM and polyps, and CT scores of those patients were highest. The sensitivity rates of fungal isolates were 97.8% for amphotericin B, caspofungin, and voriconazole; 74.4% for itraconazole; and 6.4% for fluconazole. The presence of EM with or without FH leaded to more extended CRS, but a part of positive fungal cultures were together with EM in patients with CRS. Sensitivity to antifungal agents, except fluconazole, was high. Because many factors can contribute to the pathogenesis of CRS, medical treatment should be considered on a case-by-case basis.

Nasal myiasis by fruit fly larvae: a case report.

We report a case of human nasal myiasis caused by fruit fly larvae in a 33-year-old man who visited the south coast of Turkey. The fruit fly (Drosophila melanogaster) is endemic in the southern part of Turkey. Infestation with fruit fly larvae must be considered in patients who are visiting areas of the world where Drosophila is endemic. This is the first reported case of fruit fly larvae causing human nasal myiasis. The clinical presentation and treatment strategies are discussed.

Diagnosis of Helicobacter pylori infection and determination of clarithromycin resistance by fluorescence in situ hybridization from formalin-fixed, paraffin-embedded gastric biopsy specimens.

A reliable diagnostic test for Helicobacter pylori is important in clinical practice and research. The ideal diagnostic test for H. pylori should be sensitive, specific, and cost-effective. Helicobacter pylori resistance to clarithromycin is a common reason for failure of eradication therapy. The aim of this study was to evaluate the fluorescent in situ hybridization (FISH) method to detect H. pylori and determine clarithromycin resistance in formalin-fixed, paraffin-embedded gastric biopsy specimens. One hundred seventeen gastric biopsy specimens from patients with dyspepsia were examined for the presence of H. pylori by conventional culture, FISH, and histopathological methods. A set of fluorescent-labeled oligonucleotide probes binding to either H. pylori 16S rRNA or 23S rRNA sequences were used for FISH analysis. Phenotypic antibiotic susceptibilities of the isolates were tested using the Epsilometer test method (E test). Helicobacter pylori was detected in 70 of 117 biopsy specimens by histopathological examination and FISH, whereas it was detected in 47 specimens by culturing. Histopathology and FISH techniques failed to identify H. pylori in 1 biopsy sample isolated by culture. Clarithromycin resistance was found in 11 of 46 H. pylori isolates using the E test method. All of the phenotypic resistance measurements of isolates were correlated with genotypic clarithromycin resistance. Eleven clarithromycin-resistant strains were identified by FISH. The diagnosis of H. pylori infection and the determination of clarithromycin resistance in formalin-fixed, paraffin-embedded specimens using FISH is promising because it provides a rapid, reliable, and culture-independent diagnosis.

Helicobacter pylori infection in hemodialysis patients: susceptibility to amoxicillin and clarithromycin.

AIM: To evaluate susceptibility of Helicobacter pylori to amoxicillin and clarithromycin in end-stage renal disease (ESRD) patients and non-uremic controls. METHODS: The subjects with dyspeptic complaints were 33 ESRD patients and 46 age- and sex-matched non-uremic controls who exhibited H pylori on antral biopsy specimens. The two groups were age and sex matched. The H pylori strains' pattern of susceptibility to amoxicillin and clarithromycin was investigated with the agar dilution technique. RESULTS: None of the H pylori strains from either group showed resistance to amoxicillin with the agar dilution method. Twelve (36.4%) of the ESRD group strains and 7 (15.2%) of the control group strains showed resistance to clarithromycin, and this difference was statistically significant (P<0.05). CONCLUSION: Resistance to amoxicillin does not appear to be an important problem in H pylori-infected ESRD and non-uremic patients in our region. In contrast, the rates of resistance to clarithromycin are high, particularly in the ESRD population.

[Clarithromycin resistance of Helicobacter pylori strains isolated from antral biopsy specimens]. / Antral biyopsi örneklerinden izole edilen Helicobacter pylori suslarinda klaritromisin direnci.

Helicobacter pylori is the main causative agent of peptic ulcer disease. Clarithromycin resistance in H. pylori is one of the common causes of failure of the eradication therapy based on the use of amoxicillin-clarithromycin and proton pump inhibitors. In this study, 78 H. pylori strains isolated from antral biopsy specimens of dyspeptic patients have been tested for susceptibility to clarithromycin, with NCCLS approved agar dilution and the E-test methods. Sixteen (20.5%) of the strains were found resistant to clarithromycin, and the resistance rates were similar obtained by E-test and agar dilution method. The high rate of clarithromycin resistance among H. pylori strains in our study population, may give a direction to follow-up of the eradication therapy and to define regional treatment policies.