Adrian Man, Cosmin Moldovan and Minodora Dobreanu
Starting with the first issue of 2013, the Romanian Review of Laboratory Medicine has implemented a new editorial and publishing system. By this editorial, we try to clarify to all the readers and authors the major changes and their outcome in the journal’s evolution. Thus, we present details related to the current internal organization of the editorial board and the editorial workflow of the submitted manuscripts.
Adrian Man, Claudia Bănescu, Minodora Dobreanu and Cornel Fraefel
Successful experiments in molecular biology require good knowledge about various methods and protocols. In molecular biology, nucleic acid manipulation is the essence, starting with the quality of extraction and ending with several analysis assays (PCR, RT-PCR, qPCR, PCR arrays, molecular cloning, etc). Though many of these are so called “standardized”, in practice there are many variables that can influence the outcome of the experiment. Due to the importance of optimal primer design in PCR assays, we will focus on primer designing and checking software, but we also present other useful free tools that can help researchers in the molecular biology field
Lidia Man, Adrian Man, Cristina Oana Mărginean, Ana Maria Pitea and Maria Despina Baghiu
Purpose: to evaluate the serum levels of micronutrients in children with nutritional disorders, and to find if there is a direct correlation between them and the anthropometric measurements. Materials and methods: the study was conducted on 125 children (0-18 years); the working group consisted in children with Z-score < -2 standard deviations for at least one anthropometric measurement, while the children without growth disorders were considered as controls. Thus, for each anthropometric measurement, we had different working/control groups that were used for the assessment of correlation with laboratory findings. We followed eight anthropometric parameters and their relation with five of the micronutrients (Ca, Mg, Fe, Zn and Cu). Results: no statistical differences were found in micronutrients serum levels between genders or provenance. Most mean serum levels of micronutrients were lower in the children with Z-scores < -2 standard deviations (except Cu). Mg and Ca were positively correlated with most of the anthropometric measurements. For Fe, Zn and Cu, we found no correlation with any of the anthropometric measures. Differences in mean serum levels were found for Mg, with lower values in children with low weight-forage and triceps-skinfold-thickness, and for Cu, with higher levels in children with low triceps-skinfold-thickness. The red blood cell indices were positively associated with Fe and Zn levels. Conclusions: correlations between the serum level of micronutrients and anthropometric evaluation scores were found for Mg and Ca, but not for Fe and Zn, which were instead directly correlated with red blood cells indices. Mg, Fe and Zn tend to present small serum values in children with growth deficits. Considering the costs, the routine evaluation of Zn and Cu serum levels in growth disorder suspicion is not justified in our geographic area.
A. Man, P.E. Muntean and Anca Mare
Aim: To present the etiology of bacteremic syndromes and antibiotic susceptibility of blood culture isolates from a Romanian county hospital, as well as their distribution within different wards.
Methods: We retrospectively analyzed the blood culture data collected from patients hospitalized in the County Emergency Clinical Hospital of Tirgu Mures over a period of two years. We followed aspects regarding the identified bacterial species, their distribution by sex, age groups and wards, the spectrum of resistance to antibiotics and main resistance phenotypes.
Results: Most positive samples came from ICU, nephrology and urology. The most isolated bacteria were coagulase-negative staphylococci, Escherichia coli, and Staphylococcus aureus. All isolates showed a high resistance to most classes of antibiotics, staphylococci being susceptible to glycopeptides, oxazolidinones and glycylcyclines, and the enterobacteria to aminoglycosides and carbapenems. The resistance in non-fermentative bacilli exceeded 80% to most classes of antibiotics. The methicillin-resistance was 36% for coagulase-negative staphylococci and 82% for Staphylococcus aureus; the percentage of extended-spectrum beta-lactamase producing strains was 30%.
Conclusions: The etiology of bacteremic syndromes is specific to the ward profile, the Staphylococcus spp. being primarily isolated from wards where invasive procedures are frequently performed, while the enterobacteria from urology and nephrology wards. The level of antibiotic resistance is higher in surgery and ICU wards, with also higher percentage of resistance phenotypes than in medical wards.
Adrian Maier, Adrian Man, Călin Chibelean, Teodora Cighir, Eniko Nemes-Nagy, Ioana Maier, Ciprian Todea, Oliver Vida and Orsolya Martha
Objective: To evaluate the bacteriological features in non-struvite nephrolithiasis and in its associated urinary tract infection, and to establish the relationship between the two pathologies.
Methods: The non-struvite calculi from 132 patients were aseptically extracted by percutaneous nephrolithotomy (PNL). The midstream urine and calculi were bacteriologically and biochemically processed.
Results: Most calculi (78%) were located to renal pelvis, associated with hydronephrosis, the biochemical composition confirming the lack of struvite and revealing the predominance of calcium oxalate. The females presented significantly more colonized calculi (50%) than males (21.9%), with higher bacteriological diversity. There is a significant relation between the presence of colonized calculi and urinary tract infections, 24.2% of calculi and 25.8% of the urine samples presenting positive cultures. In 70.4% of cases, we found the same antibiotic resistance pattern between the pathogens isolated from calculi and urine, thus considering them identical strains. The Enterobacteriaceae represented the most predominant bacteria both from calculi (62.5%) and urine (63.6%), approximatively 30% being resistant to cephalosporins and over 50% resistant to fluoroquinolones, ampicillin and tetracycline. There were 3.8% of cases in which the calculi were colonized but the urine was sterile, the bacteria being sensitive to cephalosporins that are used as prophylaxis.
Conclusions: In all the cases, the same bacterial species was found both in calculi and urine, and 70.4% of them were phenotypically identical. The resistance to the second generation cephalosporins is lower than in the case of other antibiotics, which makes them the most suitable for prophylaxis in PNL.
Cecilia Petrovan, Despina Luciana Bereczki-Temistocle and Adrian Man
Background: Candida, and especially Candida albicans, colonizes the oral mucosa and becomes invasive when the immune system weakens. Therefore, frequently, oral and maxillofacial pathology can be associated with Candida. Objective: The qualitative and quantitative assessment of Candida colonization of the oral cavity in patients with oro-maxillo-facial conditions; to establish an association between the different contributing factors and colonization. Material and methods: Samples were collected from 70 patients hospitalized in the Clinic of Oral and Maxillo-Facial Surgery, by rinsing the mouth with sterile saline; historical data were also collected. The samples were analyzed at the Microbiology Laboratory of the University of Medicine and Pharmacy of Tîrgu Mureș, using Sabouraud agar medium. For each isolate, the antifungigram for Fluconazole and Voriconazole was performed following the CLSI standards. Results: From the 70 samples 45.7% were positive for 11 different yeast species. Regarding predisposing factors, most importantly, smoking was significantly associated with Candida colonization (OR = 2.34; 95% CI = 1.42-3.84; p <0.05). Other factors, such as radiotherapy, oral hygiene or antibiotics, are related, but not statistically significant in our study. Candida albicans was the predominant species (38.8%). The testing of Candida albicans and non-albicans to Fluconazole showed an increased resistance (52.4%) in both cases, while the resistance to Voriconazole was 50% and 12.5%, respectively. Conclusions: Colonization of the oral cavity with Candida is present in about half of the patients with OMF conditions, and this is probably not only due to classical predisposing factors, but also due to chronic oral pathology and to several risk factors like smoking or radiotherapy.
Adrian Man, Andrei Şerban Gâz, Anca Delia Mare and Lavinia Berţa
Alcohol based solutions are among the most convenient and wide spread aid in the prevention of nosocomial infections. The current study followed the efficacy of several types and isomers of alcohols on different bacterial species. Seven alcohols (ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl alcohol, and ethylene glycol) were used to evaluate their minimal inhibitory and bactericidal effects by microdilution method on bacteria that express many phenotypical characteristics: different cell-wall structure (Gram positive/negative bacteria), capsule production (Klebsiella pneumoniae), antibiotic resistance (MRSA vs MSSA) or high environmental adaptability (Pseudomonas aeruginosa). Results: The best inhibitory effect was noticed for n-propyl, followed by iso-propyl, n-butyl, and iso-butyl alcohols with equal values. Ethylene glycol was the most inefficient alcohol on all bacteria. In K. pneumoniae and P. aeruginosa, the bactericidal concentrations were higher than the inhibitory one, and to a level similar to that encountered for most of the Gram-positive bacteria. Among Gram-positive cocci, E. faecalis presented the lowest susceptibility to alcohols. Conclusions: All alcohols presented good effect on bacteria, even in low concentrations. Compared to ethanol as standard, there are better alternatives that can be used as antimicrobials, namely longer-chain alcohols such as propyl or butyric alcohols and their iso- isomers. Ethylene glycol should be avoided, due to its toxicity hazard and low antimicrobial efficacy. Bacterial phenotype (highly adaptable bacteria, biofilm formation) and structure (cell wall structure, presence of capsule) may drastically affect the responsiveness to the antimicrobial activity of alcohols, leading to higher bactericidal than inhibitory concentrations.
Lavinia Grama, Adrian Man, Daniela-Lucia Muntean, Șerban Andrei Gâz Florea, Francisc Boda and Augustin Curticăpean
Polyoxometalates are important inorganic compounds with a broad range of pharmacological properties, including antiviral, antibacterial, antiprotozoal or antitumoral activities, even that their molecular mechanism of action is poorly understood. Purpose: In this paper we evaluated the antibacterial activity of some saturated polyoxotungstates (POW) compounds, since nowadays, the increasing resistance of bacteria to drugs represents a major health problem. Materials and methods: The antibacterial activity was studied by disk diffusion method as a possible screening method and by successive micro-dilutions method. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) have been calculated for each compound by successive dilutions. We also compared the reliability of each testing method for this particular POW evaluation. Results: The best antibacterial activity was expressed by H4[SiW12O40]*nH2O and the lowest by Na3[PW12O40]*nH2O, but with very good activity on Staphylococcus spp., especially on MRSA. The POW activity occurs only at relatively high concentrations, and it is dependent on bacterial strain, with very good activity on Staphylococcus spp. The most reliable method for assessing the antibacterial effects of POW is micro-dilutions. POWs could be practically applied in hospital decontamination and could have a possible in vivo antibacterial application.
Şerban Andrei Gâz Florea, Diana Ciurca, Anca Mare, Adrian Man, Bogdan Cordoş, Anda-Lavinia Grama and Daniela-Lucia Muntean
Background: Snake venom is a complex mixture of biologically active substances. Some peptides and proteins from snake have already demonstrated their therapeutically potential. The venom of Naja haje, an Elapidae member, has been analyzed from this point of view. Understanding the fully biochemical role of its enzymes has determined the scientists to find new separation and identification methods.
Objective: Our goal was to develop an optimal HPLC analytical method for separation and identification of Naja haje snake venom components, known for its neurotoxic activity. In addition, we wanted to find out if crude snake venom could inhibit the development of both Gram-positive and Gram-negative bacterial cultures. Materials and Method: Analysis of venom was performed on a HPLC system using a C16 column with UV detection at 210 nm. The analysis was done using two mobile phases, containing different concentrations of acetonitrile and trifluoroacetic acid aqueous solution at different pH values. An elution gradient was set at a flow of 0.60 mL/min. Bactericidal activity was quantified by measuring inhibition diameter around an aseptically disk filled with crude venom using Staphylococcus aureus and Escherichia coli. Results: An optimal HPLC analytical method has been developed by changing different parameters such as the pH value of mobile phase A or the elution gradient. The best resolution were obtained at a pH value of 7.4, in gradient varying from 5% to 45% in mobile phase B. Microbiological studies of the venom showed that Gram-positive bacteria growth was inhibited by crude venom, while on Gram-negative bacteria growth no effect was observed. Inhibition zone is dose-dependent and fresh crude venom is with 30% more potent than venom freeze and kept at -55°C. Conclusions: A comprehensive catalog of venom composition may serve as a starting point for studying structurefunction correlations of individual toxins for the development of new research tools and drugs of potential clinical use.