Comparison of nested, multiplex, qPCR; FISH; SeptiFast and blood culture methods in detection and identification of bacteria and fungi in blood of patients with sepsis

BACKGROUND: Microbiological diagnosis of sepsis relies primarily on blood culture data. This study compares four diagnostic methods, i.e. those developed by us: nested, multiplex, qPCR (qPCR) and FISH with commercial methods: SeptiFast (Roche) (SF) and BacT/ALERT® 3D blood culture system (bioMérieux). Blood samples were derived from adult patients with clinical symptoms of sepsis, according to SIRS criteria, hospitalized in the Intensive Care Unit. RESULTS: Using qPCR, FISH, SF, and culture, microbial presence was found in 71.8%, 29.6%, 25.3%, and 36.6% of samples, respectively. It was demonstrated that qPCR was significantly more likely to detect microorganisms than the remaining methods; qPCR confirmed the results obtained with the SF kit in all cases wherein bacteria were detected with simultaneous confirmation of Gram-typing. All data collected through the FISH method were corroborated by qPCR. CONCLUSIONS: The qPCR and FISH methods described in this study may constitute alternatives to blood culture and to the few existing commercial molecular assays since they enable the detection of the majority of microbial species, and the qPCR method allows their identification in a higher number of samples than the SF test. FISH made it possible to show the presence of microbes in a blood sample even before its culture

New arylpiperazine derivatives with antidepressant-like activity containing isonicotinic and picolinic nuclei: evidence for serotonergic system involvement

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Pharmacological and Structure-Activity Relationship Evaluation of 4-aryl-1-Diphenylacetyl(thio)semicarbazides

This article describes the synthesis of six 4-aryl-(thio)semicarbazides (series a and b) linked with diphenylacetyl moiety along with their pharmacological evaluation on the central nervous system in mice and computational studies, including conformational analysis and electrostatic properties. All thiosemicarbazides (series b) were found to exhibit strong antinociceptive activity in the behavioural model. Among them, compound 1-diphenylacetyl-4-(4-methylphenyl)thiosemicarbazide 1b was found to be the most potentan algesic agent, whose activity is connected with the opioid system. For compounds from series a significant anti-serotonergic effect, especially for compound 1-diphenylacetyl-4- (4-methoxyphenyl)semicarbazide 2b was observed. The computational studies strongly support the obtained results

Nutritional value and coagulation properties of milk from local cow breeds, including the selected macro- and micronutrients and trace elements

Cilj ovog istraživanja bio je odrediti prehrambenu vrijednost mlijeka na osnovu kemijskog sastava i sadržaja odabranih mineralnih tvari, te sposobnost koagulacije sirovog mlijeka izvornih pasmina - poljsko crvenih i bijelo-crnih, lokalno uzgojenih simentalskih krava, uzimajući u obzir utjecaj sezone. Istraživanje je provedeno na ukupno 418 uzoraka mlijeka uzetih tijekom proljeća/ljeta (travanj do rujan) odnosno jeseni/zime (listopad do ožujak). Uzorcima su određivani kemijski sastav, energijska vrijednost, sposobnost koagulacije (RCT - vrijeme zgrušavanja, A30 - čvrstoća gruša, K20 - vrijeme potrebno za postizanje odgovarajuće čvrstoće gruša), prinos sira te sadržaj K, Na, Ca, Mg, Zn, Fe, Mn, Cu, Ni i Co. Također je određeno u kojem postotku konzumacija 250 mL testiranih uzoraka mlijeka pokriva dnevni preporučeni unos Ca i Mg za djecu, adolescente i odrasle. Mlijeko pasmine poljsko crveno sadržavalo je najviše koncentracije osnovnih nutrijenata (1 kg mlijeka imalo je energijsku vrijednost od 823,98 kcal) i imalo je najbolju sposobnost koagulacije mlijeka (najkraće vrijeme zgrušavanja (16:09 min, p≤0,01) i najčvršći gruš (30,74 mm)). Ovaj uzorak mlijeka imao je bolji nutritivni profil budući da je sadržavao najviši postotak preporučenih dnevnih unosa za Ca i Mg. Sirovo mlijeko dobiveno u sezoni ispaše imalo je bolje parametre sposobnosti koagulacije. Sadržaj mineralnih tvari nije bio uvjetovan pasminom ni sezonom proizvodnje. To je vjerojatno posljedica većih udjela ovih nutrijenata u stočnoj hrani, što je povezano s njihovim sadržajem u tlu kao i sa stupnjem fertiliziacije.The aim of the study was to assess the nutritional value of milk based on its chemical composition and content of selected minerals and to determine the coagulation properties of raw milk from native Polish breeds, i.e. the Polish Red and the White-backed, as well as locally raised Simmental cows, taking into account the production season. The research material consisted of 418 milk samples taken during the spring/summer season (April to September) and autumn/winter season (October to March). The following parameters were determined in the samples: chemical composition, energetic value, coagulation properties (RCT - rennet clotting time, A30 - curd firmness, K20 - curd firming time), cheese yield, and content of K, Na, Ca, Mg, Zn, Fe, Mn, Cu, Ni and Co. The percentage of the daily allowance for Ca and Mg in children, adolescents and adults from 250 mL of milk was determined. The milk of Polish Red cows had the highest concentration of basic nutrients (1 kg of milk provided 823.98 kcal) and the best coagulation parameters (the shortest clotting time (16:09 min, p≤0.01) and firmest curd (30.74 mm). This milk had a more beneficial nutritional profile, as it contained the highest percentage of the recommended daily allowances of Ca and Mg among all groups. The raw milk obtained in the pasture season showed more favourable coagulation properties. The content of minerals in milk is not clearly determined by the cow breed or the season. It is probably due to the higher levels of these nutrients in the feed, which in turn results from their content in the soil and the degree of fertilization

Coagulase-negative staphylococci contained in gut microbiota as a primary source of sepsis in low- and very low birth weight neonates

Background: There are only a few reports in the literature about translocation of coagulase-negative staphylococci (CoNS) as a primary cause of sepsis in neonates, although CoNS are among a short list of “translocating” bacteria when present in abundance. Methods: 468 blood samples, 119 stool samples, and 8 catheter tips, from 311 neonates, were tested for presence of microorganisms. CoNS strains isolated from the blood and stool or from blood and catheter tip of the same newborn at approximately the same time were paired and typed with PFGE (Pulse-Field Gel Electrophoresis) method. The strains were then tested for the presence of adherence genes and biofilm formation. Results: The strains with identical PFGE profiles in comparison to those with non-identical profiles differed in terms of the pattern of the virulence genes and showed a lack of the genes related to adherence, but more often presence of IS256, which is related to virulence. They also were phenotypically unable to adhere to intestinal Caco2 cells. Conclusions: A considerable proportion of CoNS strains isolated from bloodstream of VLBW/LWB neonates was identical to the strains isolated from faeces of the same neonates at the same time. These observations may offer indirect evidence indicating that at least some CoNS can translocate from the gastrointestinal tract of the premature neonates into the bloodstream and thus cause generalized infection