The environmental monitoring in hospital pharmacy cleanroom and microbiota catalogue preparation

Knowing the normal cleanroom microbiota is the basis for ensuring microbiological quality; assess changes and the introduction of new sampling methods. During our study, we prepared a catalogue of cleanroom microorganisms located in four different cleanrooms in University Clinical Centre Ljubljana Pharmacy. Catalogue is prepared as a basis for assessing the suitability of the new rapid microbiological method and subsequent correlation of the results of both methods. The results of our study showed that 78% of isolated bacteria are Gram-positive. However, in more than 70% isolated bacteria were the part of the normal human microbiota, 10–15% of the microorganisms originated from the air, mainly spore-forming bacteria of the genus Bacillus and fungi, and 5–10% of the Gram-negative microorganisms that originated from the water and represent the potential endotoxins (pyrogens).     

1H−15N HMBC NMR as a tool for rapid identification of isomeric azaindoles: The case of 5F‐MDMB‐P7AICA

The high frequency of the synthetic cannabinoid receptor agonists (SCRAs) emergence renders this group of new psychoactive compounds particularly demanding in terms of detection, identification, and responding. Without the available reference material, one of the specific problems is differentiation and structure elucidation of constitutional isomers. Herein, we report a simple and efficient flow chart diagram applicable for a rapid nuclear magnetic resonance (NMR) identification and differentiation between azaindoles, 4‐, 5‐, 6‐, and 7‐azaindole, which is a common structural motif of synthetic cannabinoids. The flow chart diagram is based on ¹H NMR and ¹H–¹⁵N NMR spectra, and to prove the concept, it has been tested on 5F‐MDMB‐P7AICA ( 1 ). Spectral and analytical data including standard 1D and 2D NMR spectra, gas chromatography?mass spectrometry (GC?MS), Fourier transform infrared?attenuated total reflectant (FTIR?ATR), Raman, melting point, and combustion analysis are provided for compound 1 .     

Tick-borne encephalitis in Slovenia: data from a questionnaire survey

There has been a marked increase in the incidence of tick-borne encephalitis (TBE) in the last two decades, with new foci of the disease evolving. TBE is endemic in Slovenia, and the incidence rate is one of the highest in the European Union. A survey of notified TBE cases started in 1999, aiming to collect additional epidemiological data that are not available through the usual notification system. From 1999 to 2009, a short, anonymous, self-administered questionnaire was sent to 2779 notified TBE cases. The response rate was 69.5%. The willingness to respond was higher in those over 60 years of age, but did not differ by gender. In all, 1564 (82.3%) of the responders had a tick bite on one or multiple sites on the body. Age and gender influenced the location of the tick bite: males patients most often had the tick located on the torso, and females had them on the legs. Ticks located on the head and neck were significantly higher in preschool children compared to school children and adults. The estimated duration of tick attachment was less than 6 h in 23.5% of TBE cases. Long attachments (more than 24 h) were reported by only 10% of the patients. The tick bite occurred while the TBE patients were engaged in leisure time activities (sports or camping, 32.8%), mushroom or berry picking (30.2%), or farming (23.3%). Almost two-thirds of TBE patients reported that they had practiced at least one of the recommended preventive measures, most frequently self-inspection, and least often repellent use. The patients were asked to disclose the most probable geographical location of the TBE viral (TBEV) infection. Maps were created on a county level and showed that the risk of TBE is widespread, with few counties unaffected. The counties located in the northern and the central areas had the highest risk of TBEV infection.     

Molecular characterization of a new porcine rotavirus P genotype found in an asymptomatic pig in Slovenia

Rotaviral RNA was detected in the stool sample of an asymptomatic fattening pig at a Slovenian pig farm. To characterize the rotavirus, RT-PCR was used, employing primers specific for the VP7, VP4 and NSP4 genes. Specific products were purified and the sequencing reaction was performed for the molecular analysis of amplified genes. Nucleotide and amino acid sequences of the VP7 gene were found highly identical (85.3-88.1% and 90.7-91.6%) to G1 genotype strains. Phylogenetic and molecular analyses of the VP7 antigen regions revealed the sample to be from a new lineage of G1 genotype. In the molecular analysis of the VP4 gene, only 70.9% nucleotide (76.2% amino acid) identity was found with the most related rotavirus VP4 gene from GenBank. Following this, the NSP4 gene was also analyzed. After the phylogenetic analysis, it clustered with the NSP4 B genotype, but also seemed to represent a new lineage of this genotype. This new rotavirus strain, named P21-5, differed greatly from all rotaviruses characterized so far in all three genes analyzed. The virulence of this strain is not clear yet and has to be investigated.     

MD-2 as the target of curcumin in the inhibition of response to LPS

Curcumin is the main constituent of the spice turmeric, used in diet and in traditional medicine, particularly across the Indian subcontinent. Anti-inflammatory activity and inhibition of LPS signaling are some of its many activities. We show that curcumin binds at submicromolar affinity to the myeloid differentiation protein 2 (MD-2), which is the LPS-binding component of the endotoxin surface receptor complex MD-2/TLR4. Fluorescence emission of curcumin increases with an absorbance maximum shift toward the blue upon the addition of MD-2, indicating the transfer of curcumin into the hydrophobic environment. Curcumin does not form a covalent bond to the free thiol group of MD-2, and C133F mutant retains the binding and inhibition by curcumin. The binding site for curcumin overlaps with the binding site for LPS. This results in the inhibition of MyD88-dependent and -independent signaling pathways of LPS signaling through TLR4, indicating that MD-2 is one of the important targets of curcumin in its suppression of the innate immune response to bacterial infection. This finding, in addition to the correlation between the dietary use of curcumin and low incidence of gastric cancer in India, may have important implications for treatment and epidemiology of chronic inflammatory diseases caused by bacterial infection.     

Short-term “double natural orifice catheterization”: Nonoperative management of hydrocolpos in persistent cloaca patients – case series

Purpose

Management of hydrocolpos in cloaca patients is of clinical importance. We report a nonoperative method for the management of hydrocolpos in the form of initial catheter decompression, followed by an endoscopy with catheters placement into both the bladder and vagina, and leaving these catheters for 9 and 14 days respectively.

Do C1q or IgM nephropathies predict disease severity in children with minimal change nephrotic syndrome?

Background

It has been suggested that C1q and immunoglobulin M (IgM) nephropathy are variants of minimal change nephrotic syndrome (MCNS). Many researchers believe that these two conditions signify a worse prognosis for children with MCNS in comparison with immunofluorescence (IF)-negative MCNS. The aim of our study was to determine the prognostic significance of C1q nephropathy and IgM nephropathy in children with MCNS.

Methods

Fifty-five children with MCNS who had been biopsied over the course of 24 years at our institution were retrospectively categorized into three groups on the basis of IF microscopy findings: IF-negative MCNS (29/55 patients), MCNS with IgM nephropathy (19/55 patients), and MCNS with C1q nephropathy (7/55 patients). Clinical characteristics at disease presentation, clinical course, and renal outcome were compared between groups during the median follow-up period of 16.9 years (minimum 1.0, maximum 31.1 years).

Results

No statistically significant differences in clinical characteristics at disease presentation, clinical course, and renal outcome were found. Children with IgM nephropathy, C1q nephropathy, and IF-negative MCNS were clinically indistinguishable.

Conclusions

We concluded that C1q or IgM nephropathy variants do not seem to signify a worse prognosis in children with MCNS in comparison with IF-negative MCNS.