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Antibiotic overuse has led to the emergence of resistant strains of bacteria, such as Methicillin-resistant Staphylococcus aureus (MRSA), which has become a global public health concern. The aim of this study was to investigate the genetic and microbiological characteristics of MRSA in hospitals in Crete, located in the southeastern Balkan region. S. Aureus samples were collected from hospitalized patients and tested for antibiotic susceptibility using the Kirby-Bauer disk diffusion method. The presence of mecA, enterotoxin genes (sem gene or seg genes), agr genes, and PVL genes was detected using PCR. Of the 141 S. Aureus strains tested, 102 (72.34%) were found to have the genes of interest, with 69 MRSA (48.9%) mecA+ strains and 33 methicillin-susceptible S. Aureus (MSSA) mecA negative strains. The most prevalent gene detected was the agr gene (48.93%), followed by the enterotoxin gene (44.68%). Four agr groups (I – IV) were detected in 69 MRSA isolates (48.9%). This study highlights the prevalence of MRSA in hospitals in Crete and emphasizes the importance of appropriate antibiotic use and infection control measures to prevent the spread of antibiotic-resistant bacteria. Although a correlation between the presence of the detected genes and certain resistance phenotypes was not observed, the findings indicate the necessity to trace these genes using PCR and involve them in clinical diseases. The circulation of sensitive strains to antibiotics and resistant strains to some antibiotics with different resistance phenotypes was also noted in the hospital. Overall, this study provides important insights into the genetic and microbiological characteristics of MRSA in the southeastern Balkan region, and the results can inform future efforts to control the spread of antibiotic-resistant bacteria.

Introduction. In the case of many patients with chronic rhinosinusitis (CRS), antibiotic and steroid therapies fail, and surgery is required. The recovery of patients after surgery equally depends on the postoperative behavior of each individual patient. The paper presents the outcomes of recovery after functional endoscopic sinus surgery (FESS) in a group of 74 patients. Methods. The study was conducted in patients undergoing surgical treatment by FESS, performed by the same surgeon. For the development of the statistical database, the clinical records were collected by the same investigator. Results. On the day of surgery and on the first postoperative day, 72.9% of patients reported facial pain, 41.8% nausea, 9.4% nasal bleeding, 8.8% vomiting. At 6 months postoperatively, 71.6% of patients estimated that they had a better quality of life than before surgery, 64.8% mentioned an improvement of olfaction as an effect, and 6.7% developed septal turbinate synechiae. Conclusions. No major complications were identified in the recovery of CRS patients after FESS. Postoperative facial pain was less well tolerated by young patients (18-35 years old). The improvement of smell and the increase of disease-specific quality of life are the most relevant results of recovery after FESS mentioned by the patients included in our study. The otorhinolaryngologist and the family doctor play an important role in the education of patients regarding the importance of treatment with mineral and thermal waters in post-FESS recovery. Keywords: chronic rhinosinusitis, endoscopic sinus surgery, FESS, rehabilitation, ERAS protocols

Kidney disease represents a burden for the health care system worldwide. As the prevalence continues to rise, discovering new biomarkers of early kidney damage has become crucial. Oxidative stress (OS) represents one of the main factors involved in the early stages of many syndromes leading to kidney damage. Therefore, it must be studied in detail. To date, many studies have focused on OS in advanced stages of acute kidney injury (AKI), with great success. The aim of the present study was to ascertain whether even mild renal function impairment can be linked to specific systemic markers of OS and systemic antioxidants in order to pinpoint certain biomarkers for early kidney damage. We used male rats (Rattus norvegicus) in which we induced kidney damage by injecting gentamicin for 7 days. Blood was collected 24 h after the last dose of gentamicin. Urea, creatinine, 3-nitrotyrosine (3-NT), nitric oxide (NO), malondialdehyde (MDA), thiols (TS), total oxidative stress (TOS), and interferon-γ (IFN-γ) were determined. In addition, for the antioxidant status we measured total antioxidant capacity (TAC) and interleukin-10 (IL-10). Our results demonstrated that the rats had mild renal impairment consistent with a pre-AKI stage due to the nephrotoxic effect of gentamicin. However, TOS, MDA and NO were significantly higher in the gentamicin group compared to the control group. In addition, TAC was higher in the control group. Hence, OS markers reach higher levels and may potentially be used as markers of kidney damage even in cases of mild renal function impairment.

The presence of SARS-CoV-2 in the middle ear reveals the etiopathogenesis of otitis media in COVID-19, as well as an epidemiological risk during otologic examination and surgical procedures in COVID-19 patients. The study included 8 deceased patients with COVID-19. Tissue samples from the middle ear were subjected to virology, histopathology, scanning (SEM) and transmission (TEM) electron microscopy investigation. Ethmoidal mucosa samples were processed for virology analyses. qPCR resulted positive for 75% of nasal mucosa samples and 50% of middle ear samples. Ct values showed lower viral loads in middle ear samples. A proportion of 66.6% patients with positive results in the nasal mucosa showed positive results in the middle ear, and the subtype analysis of the complete genome sequences indicated B.1.1.7 lineage for all samples. In histopathological and SEM samples, no pathological aspects were identified. TEM revealed on the background of death critical alteration of cellular morphology, suggestive structures resembling SARS-CoV-2, goblet cells and immune cells. SARS-CoV-2 can be present in the middle ear of COVID-19 patients even if there is not clinical evidence of acute otitis media. Otolaryngologists could be particularly exposed to COVID-19 infection.

(1) Background: Chronic rhinosinusitis with nasal polyps (CRSwNP) is one of the most studied rhinological disorders. Modifications of the respiratory nasal mucosa in COVID-19 patients are so far unknown. This paper presents a comparative morphological characterization of the respiratory nasal mucosa in CRSwNP versus COVID-19 and tissue interleukin (IL)-33 concentration. (2) Methods: We analyzed CRSwNP and COVID-19 samples through histopathology, scanning and transmission electron microscopy and performed proteomic determination of IL-33. (3) Results: Histopathologically, stromal edema (p &lt, 0.0001) and basal membrane thickening (p = 0.0768) were found more frequently in CRSwNP than in COVID-19. Inflammatory infiltrate was mainly eosinophil-dominant in CRSwNP and lymphocyte-dominant in COVID-19 (p = 0.3666). A viral cytopathic effect was identified in COVID-19. Scanning electron microscopy detected biofilms only in CRSwNP, while most COVID-19 samples showed microbial aggregates (p = 0.0148) and immune cells (p = 0.1452). Transmission electron microscopy of CRSwNP samples identified biofilms, mucous cell hyperplasia (p = 0.0011), eosinophils, fibrocytes, mastocytes, and collagen fibers. Extracellular suggestive structures for SARS-CoV-2 and multiple Golgi apparatus in epithelial cells were detected in COVID-19 samples. The tissue IL-33 concentration in CRSwNP (210.0 pg/7 μg total protein) was higher than in COVID-19 (52.77 pg/7 μg total protein) (p &lt, 0.0001), also suggesting a different inflammatory pattern. (4) Conclusions: The inflammatory pattern is different in each of these disorders. Results suggested the presence of nasal dysbiosis in both conditions, which could be a determining factor in CRSwNP and a secondary factor in COVID-19.

(1) Background: Chronic rhinosinusitis (CRS) represents a wide range of infectious-inflammatory processes affecting, simultaneously, the nose and paranasal sinuses mucosa. The paper presents outcomes of the investigation of CRS microbiological characteristics in a group of 32 patients. (2) Methods: The purulent samples were collected during functional endoscopic sinus surgery. Agar plates were incubated and examined. All types of colonies were identified using Matrix-Assisted Laser Desorption - Ionisation-Time of Flight Mass Spectrometry (MALDI-TOF MS). For scanning electron microscopy, samples were fixed and sputter-coated with 10 nm gold and analyzed using a scanning electron microscope. For transmission electron microscopy, samples were fixed, postfixed, and dehydrated. After polymerization, ultrathin sections were collected on carbon coated copper grids and analyzed with Jeol JEM1010 TEM. (3) Results: Positive microbiological diagnosis was obtained in 62.5% of cases. The most frequent species found are Staphylococcus aureus and Streptococcus constellatus subsp. pharyngis. Corynebacterium aurimucosum and Eggerthia catenaformis were unreported species in CRS until the present. Biofilm was evidenced in 43.7% of sinus mucosa samples. Ciliary disorientation, atrophy, and no ciliated cells were also identified. (4) Conclusion: The microbial factor—pathogen or opportunistic—is one of the most important pathological links in chronic rhinosinusitis. MALDI-TOF MS allows easily and quickly identification of germs.

Objectives. The aim of this study is to provide information about prevalence, etiology, risk factors, clinical characteristics and endoscopic features of various types of infectious esophagitis in children. Methods. We performed a total of 520 upper gastrointestinal tract endoscopies in Pediatric Clinic II, Emergency Hospital for Children, Cluj-Napoca. Indications for endoscopy in our cohort were gastrointestinal tract symptoms such as dysphagia, heartburn, or appetite loss. Results. The prevalence of infectious esophagitis in the study population was 2.11% (11 patients). Candida albicans (C. albicans) was the most frequent cause. Our data illustrates that herpes simplex virus (HSV)-induced esophagitis is common in immunocompromised patients and should be systematically suspected in cases of severe dysphagia, heartburn, or hematemesis. In the present study, all cytomegalovirus (CMV) esophagitis patients were immunocompromised. Immunodeficiency (81.8%) and prolonged antibiotic therapy with broad-spectrum antibiotics were by far the most important risk factors involved in the pathogenicity of the disease. Dysphagia, appetite loss, heartburn, epigastralgia, and hematemesis were the main clinical manifestations. Infectious esophagitis was associated with significant mortality. In four patients, endoscopy during life showed signs of infectious esophagitis, however, the precise etiology was only established post-mortem, in the pathological anatomy laboratory department. A risk factor involved in pathogenesis of post-mortem diagnosed infectious esophagitis is the DiGeorge syndrome for CMV and HSV patients. Conclusions. The study illustrates that infectious esophagitis should be considered in immunocompromised infants with prolonged antibiotic therapy with broad-spectrum antibiotics.

The article presents a synthetic molecular characterization of the methicillin-resistant Staphylococcus aureus and describes the most important community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) clones that circulate nowadays in the world: the main molecular and epidemiological characteristics, as well as notions related to the clinic of infections produced by these clones and their antibiotic resistance spectrum.The predominant clone of CA-MRSA in North America is USA300 – ST8-IV in North America, in Australia – Queensland (Qld) MRSA (ST93-IV), in Europe – ST80-IV, in Asia there is a high heterogeneity of clones population, in Africa the distribution of CA-MRSA clones is unclear, and in South America – USA 1100 and USA300-Latin American variant are predominant .The molecular diagnosis is performed by highly specialized institutions. The knowledge of clones allows the study of antibiotic resistance spectrum for each one, a fact of great importance for medical practice. Molecular epidemiology of the CA-MRSA shows that lowly restricted sales of antibiotics in shops and pharmacies, as well as medical prescribing practices without a laboratory investigation, especially in Eastern Europe and Asia, contribute to the development of new MRSA clones with increased resistance to antibiotics.

Microbiological and electrochemical assays, applying the cylinder-plate and differential pulse voltammetry as techniques, are reported for the quantitative determination of roxithromycin in serum and solid pharmaceutical form. The microbiological assay is based upon the inhibitory effect of this drug on the strain Bacillus subtilis ATCC 9372 used as the test microorganism. Linearity of the calibration curve was observed over the concentration range of 8.37–83.70 μg mL−1, with relative standard deviation values less than 5.0%. The electrochemical behavior of roxithromycin was studied at a graphite screen-printed electrode modified with graphene by using cyclic voltammetry and differential pulse voltammetry. The current value of the oxidative peak obtained for roxithromycin at 0.65 V vs. Ag/AgCl in 0.03 mol L−1 phosphate buffer solution (pH 7.0) with a scan rate of 0.1 V-1 is a linear function of the concentration in a range of 4.19–83.70 μg mL−1 (5–100 μmol L−1). A comparative study was carried out and both methods were applied for the determination of roxithromycin in solid dosage forms and spiked serum. The bioassay results of human serum samples were in accordance with the electrochemical ones (R2 = 0.988, P

Background: Establishing the diagnosis of COVID-19 and Pneumocystis jirovecii pulmonary coinfection is difficult due to clinical and radiological similarities that exist between the two disorders. For the moment, fungal coinfections are underestimated in COVID-19 patients. Case presentation: We report the case of a 52-year-old male patient, who presented to the emergency department for severe dyspnea and died 17 h later. The RT-PCR test performed at his admission was negative for SARS-CoV-2. Retesting of lung fragments collected during autopsy revealed a positive result for SARS-CoV-2. Histopathological examination showed preexisting lesions, due to comorbidities, as well as recent lesions: massive lung thromboses, alveolar exudate rich in foam cells, suprapleural and intra-alveolar Pneumocystis jirovecii cystic forms, and bilateral adrenal hemorrhage. The existing coinfection was identified after the autopsy. Establishing the diagnosis of this coinfection is difficult due to clinical and radiological similarities that exist between the two disorders. For the moment, fungal coinfections are underestimated in COVID-19 patients. Conclusion: COVID-19 and P. jirovecii coinfection should be considered, particularly in critically ill patients, and we recommend the systematic search for P. jirovecii in respiratory samples.

The first kidney transplantation was performed in 1951 and ever since then living donor transplantation became a more and more important solution for patients with end-stage renal disease (ESRD).Renal transplantation is a life-saving procedure. Morbidity and mortality on waiting-lists are strongly correlated with the time of dialysis and end-stage renal disease is one of the most important causes of death; this is the reason why transplantation has to be performed as soon as possible in order to reduce the time of dialysis.Once the transplantation is performed, a number of complications may occur in post-transplant evolution, the most important of which is rejection.The rejection may appear through several mechanisms, but one of the most frequent causes of rejection is cytomegalovirus (CMV) infection. It is very important to have a precocious and fast diagnosis of CMV infection in order to maintain the functionality and survival of the graft. PP65 CMV antigenemia has proven its effectiveness in detecting and monitoring the CMV infection in transplanted patients.In the laboratory of the Clinical Institute of Urology and Renal Transplantation (ICUTR) of Cluj Napoca the CMV infection is evidenced by two methods: PP65antigenemia and IgM antibody identification by chemiluminiscence.

Introduction: Nosocomial infections caused by Pseudomonas aeruginosa producing carbapenemases represent an important cause of morbidity and mortality among immunosuppressed patients. The aim of our study was to detect the production of metallo-carbapenemases (MBLs) by phenotypic methods and to detect the presence of the MBLs encoding genes (blaIMP and blaVIM) by PCR in P. aeruginosa strains isolated from hospitalized patients to the Regional Institute of Gastroenterology and Hepatology, Cluj-Napoca. Material and methods: Between September 2014-February 2015, we tested thirty-eight P. aeruginosa strains resistant to carbapenems according to CLSI 2014 breakpoints, determined by Vitek®2(BioMérieux),isolated from various clinical specimens. Phenotypic detection of the MBLs production was performed using the KPC/MBL Confirmation kit (ROSCO®) and the MBL Etest® IP/IPI (BioMérieux). We used the PCR method for detecting MBLs encoding genes: blaIMP, blaVIM. Results: The strains were obtained from surgery (55.3%), ICU (15.8%) and gastroenterology wards (28.9%), isolated from pus (25.8%), tracheal secretion (22.7%), bile (13.6%), sputum (10.6%), blood (10.6%), other secretions (16.7%). These strains were resistant to multiple classes of antibiotics. By ROSCO® method 28/38 strains (73.7%) were positive with imipenem ± dipicolinic acid (DPA) and 22/38 (57.9%) with meropenem ± DPA. Etest® waspositive for the 28/38 strains (73.7%). 11 strains (28.9%) were positive for KPC with the screening method. We identified: 6 blaIMP+ (15.8%), 2 (5.3%) blaVIM+ and 4 blaIMP+/blaVIM+ strains (10.5%). Conclusion: Both genes encoding MBL were found, alone or in combination. The increasing level of carbapenem resistance of these strains impose their routine testing to detect MBL.

The first aim of the study was to compare the antibacterial activity of several types of honey of different origins, against some bacterial resistant strains. The strains had been isolated from patients. The second aim was to discover the correlations between the antibacterial character of honey and the physico-chemical properties of the honey. Ten honey samples (polyfloral, linden, acacia, manna, and sunflower) from the centre of Romania were tested to determine their antibacterial properties against the following bacterial species: Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Salmonella enterica serovar Typhimurium, Bacillus cereus, Bacillus subtilis, and Listeria monocytogenes. Bacterial cultures in nutrient broth and the culture medium Mueller-Hinton agar were used. The susceptibility to antibiotics was performed using the disk diffusion method. All honey samples showed antibacterial activity on the isolated bacterial strains, in particular polyfloral (inhibition zone 13-21 mm in diameter) - because it is the source of several plants, and manna (inhibition zone 13-19.5 mm in diameter), and sunflower (inhibition zone 14-18.5 mm in diameter). Pure honey has a significant antibacterial activity against some bacteria which are resistant to antibiotics. Bacterial strains differed in their sensitivity to honeys. Pseudomonas aeruginosa and Staphylococcus aureus were the most sensitive. The present study revealed that honey antibacterial activity depends on the origin of the honey. We also found that there was a significant correlation between antibacterial activity of honeys and the colour of the honey but not between acidity and pH. The statistical analysis showed that the honey type influences the antibacterial activity (diameter of the bacterial strains inhibition zones).

Transplantation activity is increasing, leading to a growing number of patients at risk for toxoplasmosis. We reviewed toxoplasmosis prevention practices, prevalence, and outcomes for hematopoietic stem cell transplant (HSCT) and solid organ transplant (SOT; heart, kidney, or liver) patients in Europe. We collected electronic data on the transplant population and prevention guidelines/regulations and clinical data on toxoplasmosis cases diagnosed during 2010-2014. Serologic pretransplant screening of allo-hematopoietic stem cell donors was performed in 80% of countries, screening of organ donors in 100%. SOT recipients were systematically screened in 6 countries. Targeted anti-Toxoplasma chemoprophylaxis was heterogeneous. A total of 87 toxoplasmosis cases were recorded (58 allo-HSCTs, 29 SOTs). The 6-month survival rate was lower among Toxoplasma-seropositive recipients and among allo-hematopoietic stem cell and liver recipients. Chemoprophylaxis improved outcomes for SOT recipients. Toxoplasmosis remains associated with high mortality rates among transplant recipients. Guidelines are urgently needed to standardize prophylactic regimens and optimize patient management.

Bolile cu transmitere sexuală (BTS) reprezintă o cauză foarte importantă a îmbolnăvirilor în lume, iar infecţiile prelungite şi netratate cu agenţii patogeni care provoacă BTS pot avea consecinţe serioase. Scopul studiului nostru a fost să evalueze distribuția a şase tipuri diferite de BTS (Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Ureaplasma urealyticum, Mycoplasma hominis şi Mycoplasma genitalium) în probe de urină recoltate de la bărbaţi. Intre aprilie 2014 şi aprilie 2015 s-a colectat primul jet de urină de la 52 de pacienţi simptomatici. S-a extras ADN-ul, s-a purificat şi s-a amplificat prin tehnica polymerase chain reaction (PCR) pentru detecţia simultană a agenţilor patogeni menţionaţi, iar pentru identificare s-a utilizat electoforeza în gel de agaroză 2% cu bromură de etidiu ca agent de colorare. Frecvența BTS în grupul luat în studiu a fost de 53.84% (28 de pacienţi), cei mai mulţi în grupa de vârstă 20-29 ani. Dintre pacienţii pozitivi, şase au prezentat infecţii multiple. S-au detectat 35 probe de ADN pozitive: 17 de C. trachomatis, 9 de U. urealyticum, 7 de N. gonorrhoeae şi 2 de M. genitalium. Aplicarea pe scară largă a sistemului bazat pe detecţia simultană a acestor şase agenţi patogeni care induc BTS poate să faciliteze diagnosticul, în special în infecţiile multiple.

Background: The optimal choice of antimicrobial therapy is an important problem in hospital environment in which the selection of resistant and virulent strains easy occurs. S. aureus and especially MRSA(methicillin-resistant S. aureus) creates difficulties in both treatment and prevention of nosocomial infections. Aim: The purpose of this study is to determine the sensitivity and the resistance to chemotherapy of staphylococci strains isolated from various pathological products. Material and Method: We identified Staphylococccus species after morphological appearance, culture properties, the production of coagulase, hemolisines and the enzyme activity. The susceptibility tests were performed on Mueller-Hinton medium according to CLSI (Clinical and Laboratory Standards Institute). Results: The strains were: MSSA (methicillin-susceptible S. aureus) (74%), MRSA (8%), MLS B (macrolides, lincosamides and type B streptogramines resistance) (12%) and MRSA and MLS B (6%). MRSA strains were more frequently isolated from sputum. MRSA associated with the MLS B strains were more frequently isolated from pus. MLS B strains were more frequently isolated from sputum and throat secretions. All S. aureus strains were susceptible to vancomycin and teicoplanin. Conclusions: All staphylococcal infections require resistance testing before treatment. MLS B shows a high prevalence among strains of S. aureus. The association between MLS B and MRSA remains a major problem in Romania.

Honey is used in food industry and medicine due to its nutritive, therapeutic and dietetic qualities. The microbiological characteristics of 10 unpasteurized honey samples of known origin, collected from Transylvania beekeepers (Romania) were determined. The antibacterial activity of these types of honey against Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Salmonella enteritidis, Salmonella anatum, Salmonella choleraesuis, Bacillus cereus, Bacillus subtilis subsp. spizizenii and Listeria monocytogenes strains was studied. The most sensitive to the antibacterial activity were the two staphylococus strains (the largest diameter of inhibition zone was 18 mm) and B. subtilis strains (13.5 mm). The strains of B. cereus, E. coli, L. monocytogenes and Salmonella spp. were found to present resistance to some of the honey samples. Manna, sunflower and polyfloral honeys presented high antibacterial activity while acacia and linden honeys had a lower activity in terms of the number of sensible strains. Statistical analysis shows that the type of strains and the type of honey have influence on the diameter of inhibition.