Isolation and identification of Pantoea agglomerans from the inflated bag with dried hop pellets stored under a modified atmosphere.

AIMS: Isolation, characterization and identification of possible microbial contaminant(s) in the inflated foil bag containing hop pellets packed and stored in a modified atmosphere. METHODS AND RESULTS: Package gas of the inflated foil bag containing hop pellets was analysed by gas chromatography. Compared with the reference modified atmosphere, containing about 16 vol.% of CO2 , the inflated bag atmosphere contained 53 vol.% CO2 , suggesting possible microbial contamination. Therefore, several standard and mineral media, with added hop pellets or hop infusion, were used for cultivation at different temperatures under an anaerobic atmosphere. Cultivation in mineral medium with hop pellets yielded a bacterial isolate that was identified by MALDI-TOF mass spectrometry and verified by partial 16S rRNA gene analysis as Pantoea agglomerans, a known plant epiphyte. CONCLUSIONS: A novel strain of P. agglomerans (designed as DBM 3696) was found to be suspicious of causing inflation of the foil bag containing dried hop pellets packed in modified atmosphere. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that P. agglomerans, probably hop epiphyte, could cause sporadic inflation of bags with hop pellets packed in modified atmosphere causing logistical problems during bags transport.

Molecular aspects of the interaction between Mason-Pfizer monkey virus matrix protein and artificial phospholipid membrane.

The Mason-Pfizer monkey virus is a type D retrovirus, which assembles its immature particles in the cytoplasm prior to their transport to the host cell membrane. The association with the membrane is mediated by the N-terminally myristoylated matrix protein. To reveal the role of particular residues which are involved in the capsid-membrane interaction, covalent labelling of arginine, lysine and tyrosine residues of the Mason-Pfizer monkey virus matrix protein bound to artificial liposomes containing 95% of phosphatidylcholine and 5% phosphatidylinositol-(4,5)-bisphosphate (PI(4,5)P2 ) was performed. The experimental results were interpreted by multiscale molecular dynamics simulations. The application of these two complementary approaches helped us to reveal that matrix protein specifically recognizes the PI(4,5)P2 molecule by the residues K20, K25, K27, K74, and Y28, while the residues K92 and K93 stabilizes the matrix protein orientation on the membrane by the interaction with another PI(4,5)P2 molecule. Residues K33, K39, K54, Y66, Y67, and K87 appear to be involved in the matrix protein oligomerization. All arginine residues remained accessible during the interaction with liposomes which indicates that they neither contribute to the interaction with membrane nor are involved in protein oligomerization. Proteins 2016; 84:1717-1727. © 2016 Wiley Periodicals, Inc.

Analysis of organic colouring and binding components in colour layer of art works.

Two methods of analysis of organic components of colour layers of art works have been tested: IR microspectroscopy of indigo, Cu-phthalocyanine, and Prussian blue, and MALDI-TOF-MS of proteinaceous binders and a protein-containing red dye. The IR spectra distortion common for smooth outer surfaces and polished cross sections of colour layer of art works is suppressed by reflectance measurement of microtome slices. The detection limit of the three blue pigments examined is approximately 0.3 wt% in reference colour layers in linseed oil binder with calcite as extender and lead white as a drying agent. The sensitivity has been sufficient to identify Prussian blue in repaints on a Gothic painting. MALDI-TOF-MS has been used to identify proteinaceous binders in two historical paintings, namely isinglass (fish glue) and rabbit glue. MALDI-TOF-MS has also been proposed for identification of an insect red dye, cochineal carmine, according to its specific protein component. The enzymatic cleavage with trypsin before MALDI-TOF-MS seems to be a very gentle and specific way of dissolution of the colour layers highly polymerised due to very long aging of old, e.g. medieval, samples.

Fast detection of phosphorylation of human pepsinogen A, human pepsinogen C and swine pepsinogen using a combination of reversed-phase high-performance liquid chromatography and capillary zone electrophoresis for peptide mapping.

A combination of reversed-phase high-performance liquid chromatography (RP-HPLC) and capillary zone electrophoresis (CZE) was used for characterization of alpha-chymotryptic digests of human pepsinogen A, human pepsinogen C (both isolated from stomach mucosa of patients suffering from gastric cancer), swine pepsinogen and their dephosphorylated forms. Combining RP-HPLC and CZE for peptide mapping allowed to detect phosphorylations in molecules of the above mentioned gastric zymogens. We have found one phosphate group in the molecule of human pepsinogen A and two phosphate groups in the molecule of human pepsinogen C. The investigated sample was obtained from stomach mucosa of a patient suffering from gastric cancer. An increased number of phosphate groups in molecules of human pepsinogen seems to be associated with gastric cancer. The developed method represent a suitable tool for studying relationships between specific phosphorylations of proteins and cancerogenesis or potentially could serve for early diagnosis of gastric cancer.

Application of reversed-phase high-performance liquid chromatography and capillary zone electrophoresis to the peptide mapping of pepsin isoenzymes.

A combination of reversed-phase high-performance liquid chromatography (RP-HPCL) and capillary zone electrophoresis (CZE) was used for the characterization of peptide maps of swine pepsin after its digestion with alpha-chymotrypsin. Peptide maps obtained by both methods were compared and five selected chromatographic peaks were identified on an electrophoreogram. The different order of peaks found in RP-HPLC compared to CZE confirmed the complementarity of these two methods. More peptide fragments were resolved by RP-HPLC, which was also found to be less sensitive to salt content in peptide mixtures, than by CZE, but only CZE was able to separate and identify phosphorylated and dephosphorylated peptide fragments of swine pepsin digest. CZE provides faster separation than RP-HPLC, however, the salts have to be removed by ultrafiltration or by RP-HPLC pre-separation prior to CZE analysis. Combined use of RP-HPLC and CZE for peptide mapping makes it possible to distinguish between the phosphorylated and dephosphorylated forms of swine pepsin. This is important from a diagnostic point of view, because pepsin phosphorylation may be associated with gastric cancer.

High performance liquid chromatography enables fast determination of ratios between individual proteolytic enzymes in human gastric mucosa.

The human gastric mucosa contains five isozymogens of pepsinogen (pepsinogens PGA1-PGA5), two isozymogens of gastricsinogen (gastricsinogens PGC6-PGC7) and zymogens of cathepsins. Ratios between some individual pepsins or pepsinogens are very important from a diagnostic point of view. The ratio of pepsinogen 3 to pepsinogen 5 is significant marker of gastric cancer and gastric ulcer. High-performance ion-exchange chromatography is an easy and fast method for determination of ratios between individual proteolytic zymogens in human gastric mucosa and thus could serve for additional diagnosis of gastric diseases.

Streptococcal pharyngitis-tonsillitis in Swiss children. Diagnosis and management.

In a prospective study, during a period of 1 year, 1,116 children over 1 year of age, with acute pharyngitis-tonsillitis, were examined by a group of Swiss pediatricians. Throat swabs were cultured for the presence of group A beta-hemolytic streptococcus, and treatment was withheld pending results in most cases. The throat swab cultures were positive in 29.6% of the cases. There were no incidences of acute glomerulonephritis, acute rheumatic fever or severe purulent local complications. The findings confirm the recommendations in the American literature concerning clinic, diagnosis and management, and they can be applied in Switzerland.

What seven pediatricians in middle Europe observed and did in their office practice from August 1979 to March 1980.

The authors present the systematization of medical documentation in pediatric practice checked with 5,721 recordings of consultations. General statistics and comparative statistics of 7 pediatric offices are shown. The system serves today as basic documentation for scientific retrieval and is planned to be integrated in an on-line medical information system providing features as patients' follow-up, indications of laboratory values, treatment, and facturation.

Short treatment of urinary tract infections in children.

111 episodes of acute urinary tract infection in children without urinary tract malformations were treated by trimethoprim-sulfamethoxazole, either during 3 days or during 7-10 days. Cure rate attained almost 100%. Recurrence rates of the first infection were similar after short treatment (29.6%) and after conventional treatment (30%). It is concluded that uncomplicated urinary tract infection in ambulatory practice may be treated safely and successfully by only 3 days of an appropriate antimicrobial.