Volume 23 (2015): Issue 3 (August 2015)

Background: Experimental models are essential for clarifying the pathogenesis of diabetes mellitus (DM). We aimed to provide an exhaustive description of clinical, biochemical, and hematologic features of rats with streptozotocin (STZ)-induced DM.

Methods: Wistar rats were assigned to control (n=14) or DM (n=17) groups. DM was induced using STZ (60 mg/kg, i.p.). If STZ failed to induce DM, rats were reinjected with a similar STZ dose. Bodyweight, 24-h food and water intake were measured weekly during 28 weeks. At the end of the study lipid profile, kidney function, and complete blood count were assessed.

Results: STZ induced DM in 58.82% of rats. The second STZ administration induced DM in 71.43% of the remaining rats. Diabetics presented progressive, but less significant bodyweight increase than controls, and higher food and water consumption. At the end of the study, diabetics presented higher white blood cells count, glucose, triglycerides, total and low-density lipoprotein cholesterol, and lower creatinine clearance than controls (all p≤0.02). No significant difference was observed between diabetics injected once and those that were reinjected, in any of the studied parameters.

Conclusions: This study provides one of the longest follow-ups of rats with STZ-induced type 1 DM, demonstrating that the STZ-diabetic rat replicates the most relevant clinical, biochemical, and hematologic features of human diabetes. The present data also indicate, for the first time, that rats with initial unsuccessful STZ administration can be safely reinjected, with outcomes similar to those seen in rats receiving a single injection.

Introduction. In recent years, determination of soluble transferrin receptor levels has been emerging as a test that can reliably indicate iron deficiency in various states, and that is non-invasive and easy to use. The aim of this study was: to determine reference values of soluble transferrin receptor concentrations in serums in our population, to examine the reliability of this method in the diagnosis of anemia due to iron deficiency and associated iron deficiency in anemia accompanying malignant hemopathies, and to identify possible limitations of the test in certain conditions.

Material and Methods. The prospective research included 86 patients with anemia: 46 patients with iron deficiency anemia, and 40 patients with malignant hemopathies. The control group consisted of 40 healthy persons aging over 18.

Results. Ferritin values were reduced in 76.1% of patients, while higher levels of soluble transferrin receptors appeared in 100% of patients with iron deficiency anemia. In patients with reduced serum ferritin levels, the soluble transferrin receptor/log ferritin index was statistically significantly higher than in patients in whom ferritin concentration was in the normal range (p <0.001). ROC analysis of patients with iron deficiency anemia showed that the soluble transferrin receptor/log ferritin index (AUC 0.977) and levels of soluble transferrin receptors (AUC 0.931) occupied the largest area under the curve. The best diagnostic parameter for detecting iron deficiency in patients with malignant hemopathies by ROC analysis is the soluble transferrin receptor/log ferritin index (AUC 0.770).

Conclusion. Soluble transferrin receptors are useful in the diagnosis of iron deficiency anemia, especially when ferritin values are not reduced. The calculation of soluble transferrin receptor/log ferritin index is even more reliable. In patients with malignant hemopathies, the associated iron deficiency could be best indicated by soluble transferrin receptor/log ferritin index.

In March 2012, a food poisoning outbreak was reported in a Romanian county, with a total number of 30 children affected. The symptoms (vomiting, diarrhea and abdominal pain), with onset within 1-2 hours after the ingestion of a particular food (milk), suggested a possible staphylococcal aetiology. An outbreak investigation was carried out, in accordance with the national surveillance methodology and 25 samples: stool (n=9), vomit (n=5), nasal swabs (n=9), and milk (n=2) were collected from the affected children, food handlers and suspected food. All isolated strains were sent to the Reference Centre for Staphylococci within the “Cantacuzino” National Institute of Research-Development for Microbiology and Immunology, Bucharest, Romania, for confirmation and further analysis. The aim of this study was to increase the reference laboratory capacity to confirm staphylococcal food poisoning (SFP) outbreak by defining the molecular basis of toxicity of Staphylococcus aureus (S. aureus) isolates and assessing their genetic relatedness. PCR methods have been used to detect 14 enterotoxin genes and the expression of some of these genes was proved by using a reverse transcription real-time PCR. Pulsed-field gel electrophoresis (PFGE) and Staphylococcus protein A coding gene sequence typing (spa typing) have been used to track the origin of the S. aureus contamination and to confirm the food poisoning outbreak.

Two enterotoxigenic S. aureus strains isolated from milk, twelve isolated from patients and two from food handlers were of the same spa- type (t902) and revealed an indistinguishable SmaI macrorestriction pattern after a PFGE analysis. All these strains harboured the same toxin genes profile, namely the enterotoxin gene cluster (egc), which strongly supports the evidence that the milk was the incriminated food vehicle of the outbreak and a food-handler was the most likely source of the staphylococcal food poisoning (SFP) incident.

Introduction: Hospital-acquired infections caused by Enterobacteriaceae producing different types of carbapenem- hydrolizing enzymes are now commonly observed and represent a great limitation for antimicrobial therapy. The purpose of the study was to evaluate the emergence of carbapenem-resistant Enterobaceriaceae among the strains isolated from hospitalized patients to the National Institute of Infectious Diseases, Bucharest (NIID) and the identification of different types of carbapenemases, using phenotypic methods.

Materials and methods: Between January - June 2014, 587 strains of Klebsiella pneumoniae, Enterobacter species and E.coli were isolated from various clinical specimens. We were included all non-susceptible strains to carbapenems, according to EUCAST 2014 clinical breakpoints, as determined by using microdilution MicroScan Panels (Siemens Healthcare Diagnostics). The modified Hodge test (MHT) was performed as phenotypic confirmatory test for carbapenemase production according to CLSI guidelines and the combination disk test (KPC, MBL , OXA-48 Confirm kit, Rosco Diagnostica) according to EUCAST guidelines.

Results: A total of 45 non-repeat Enterobaceriaceae (32 strains Klebsiella pneumoniae, 5 strains E.coli, 8 strains Enterobacter spp) were identified as non-susceptibile to one or more carbapenems (93,33% ertapenem, 53,33% meropenem, 48,88% imipenem). Most strains were isolated from urine (75,55%). MHT was positive in 55,6% (25/45) of carbapenem-resistant strains; in 24 cases the carbapenem-hydrolizing enzyme was identified as: OXA-48-like (n=16), KPC (n=4), MBL (n=1), KPC + MBL (n=2) and MBL + OXA-48-like (n=1). All carbapenemase- positive strains were 100% resistant to 3rd and 4th generation cephalosporins, showing less resistance to tigecycline (12,5% resistant and 25% intermediate), colistin (37,5%) and fosfomycin (41,6%).

Conclusion: During 6 months period, there were isolated 7,66% (45/587) carbapenem-resistant Enterobacteriaceae (K. pneumoniae 21,47%, E. coli 1,23%). Twenty four strains were carbapenemase-producers. The most frequent carbapenemase isolated in our study was OXA-48-like.

Background: Soluble CD14 subtype (sCD14-ST), also named presepsin, has been proposed as a novel biomarker for the diagnosis of sepsis. We hypothesized that presepsin value might be helpful in the diagnosis of infective endocarditis (IE).

Material and methods: In this prospective study a total of 29 patients with clinical suspicion of IE were enrolled. The plasma presepsin samples were collected at the admittance in the same time with blood cultures, CRP (C-reactive protein) and routine blood tests. Data about the antibiotic treatment prior to admittance were recorded. The diagnosis of IE was made using the Duke modified criteria. Receiver operating characteristic (ROC) curves analysis and binary logistic regression were performed using SPSS software, version 18. A p value less than 0.05 is considered statistically significant.

Results: Patients were divided in two subgroups: 14 patients with definite IE and 8 with IE - rejected according to the modified Duke criteria. 7 patients with final diagnosis of sepsis were excluded. Presepsin levels in patient with definite IE were significantly higher than in those with rejected IE (p<0.03). The area under the receiver operating characteristic curve (AUC) was 0.781 (95 % confidence interval (CI) 0.590 - 0.973). The threshold value of presepsin in predicting IE was determined to be 345 pg/ml, of which the clinical sensitivity and specificity were 64% and respectively, 88%. The AUC for CRP was 0.656 (95% CI 0.37-0.88).

Conclusion: Presepsin might be a useful additional diagnostic marker in patients with suspected IE. These preliminary results needs confirmation by future studies.

Objectives: Our study aimed to evaluate and compare the accuracy of C-reactive protein, Procalcitonin and Interleukine-6 in identifying serious bacterial infections (SBI) in children with fever without source.

Methods: 139 children, aged 7 days to 36 months, addressing the Emergency Department from a Romanian university hospital, were prospectively enrolled during 2013. C-reactive protein, Procalcitonin and Interleukin-6 were determined for every patient. SBI diagnosis was based on cultures results and chest radiographs.

Results: 31 patients (22.3%) had SBI. C-reactive protein [AUC: 0.87 (95%CI: 0.81-0.92)] and Procalcitonin [AUC: 0.83 (95%CI: 0.76-0.89)] proved strong prediction value for SBI and performed better than Interleukin-6 [AUC: 0.77 (95%CI: 0.69-0.84)]. For the group of children with the duration of fever less than 8 hours, Interleukin- 6 was the best predictor [AUC: 0.88 (0.76-0.95)].

Conclusions: Both C-reactive protein and Procalcitonin are strong and similar predictors for SBI, and Interleukin- 6 might be a better SBI screening tool for children with shorter duration of fever.

Sexually transmitted diseases (STDs) are a very important cause of illness worldwide and prolonged, untreated infections with STD pathogens may have serious consequences. Our study aims to evaluate the distribution of six different STDs (Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium) in male urine samples. First void urine samples from 52 symptomatic patients were collected between April 2014 and April 2015. DNA was extracted, purified and amplified via multiplex polymerase chain reaction (PCR) for the detection of the six STD pathogens, further identified using a 2% agarose gel electrophoresis with ethidium bromide as staining agent. STD frequency in the study group was 53.84 % (28 patients), mostly in the 20-29 years age group. Among positive patients, six presented multiple infections. 35 positive DNA samples were identified in the study: 17 of C. trachomatis, 9 of U. urealyticum, 7 of N. gonorrhoeae and 2 of M. genitalium. Wide scale application of the system based on the simultaneous detection of these six pathogens inducing STD may facilitate diagnosis, especially in multiple infections.

Morganella morganii (M. morganii) is a Gram-negative aerobic and facultative anaerobic rod, belonging to the Enterobacteriaceae family. This pathogen is uncommon in community-acquired infections, most often being found in postoperative nosocomial and urinary tract infections. Infection of the central nervous system with this pathogen is rare. We present the case of a 66-year-old patient who underwent colon cancer surgery, chemotherapy and radiotherapy, had left iliac anus, type 2 diabetes and developed acute meningoencephalitis caused by M. morganii. Cerebrospinal fluid examination revealed increased number of polymorphonuclear neutrophils, modified biochemistry and AmpC beta-lactamase producing M. morganii strain. After initiation of antibiotic treatment, initially with empirical therapy represented by meropenem and vancomycin, afterwards adjusted to meropenem and ciprofloxacin, according to the stain’s susceptibility to antimicrobials the patient’s evolution was favourable, in spite of the existence of two immune suppressing conditions.

We report a male infant diagnosed at the age of 10 months with hyper-IgM syndrome (HIGM) in context of severe infections caused by Streptococcus pneumoniae, Staphylococcus aureus and Candida albicans. In patient’s outcome, in spite of immunoglobulin therapy, he continues presenting bilateral suppurative otitis media due to both Candida and penicillin-resistant pneumococcus and forearm abscess caused by Staphylococcus aureus. The infant developed bilateral cataracts, chronic hepatitis and comminuted fracture secondary to bone demineralization. The patient didn’t develop opportunistic infections as compare to CD40 Ligand deficiency patients. In contrast with the majority of HIGM cases, the infant necessity for immunoglobulin substitution was very limited. As a particularity of immunological phenotype, the patient IgM value progressively increased at a high level.