Pojava i određivanje Thermoanaerobacterium i Thermoanaerobacter u konzerviranoj hrani u limenkama

In order to determine the reason for loss of vacuum in canned food, obligately anaerobic, spore forming thermophilic organisms were isolated from shelf-stable canned food containing vegetables, noodles and potatoes as main ingredients. Thermophilic bacteria from 44 canned food samples that had been stored under anaerobic conditions at 37 °C for at least 7 days were isolated. In addition, organic fertilizer used for the cultivation of some of the foods’ ingredients was examined and anaerobic, thermophilic bacteria could also be isolated from this source. Identification of bacterial strains was carried out by partial and complete 16S-rRNA-gene sequencing. Some of the obtained gene sequences showed a high level of similarity to existing 16S-rRNA gene sequences towards strains of the genera Thermoanaerobacter, Thermoanaerobium and Thermoanaerobacterium respectively, which have not yet been reported to be of importance as food spoilers. In the course of identification of these thermophilic bacteria we developed genera specific PCR-based approaches for detecting isolates belonging to the genera Thermoanaeroacterium and Thermoanaerobacter. Direct capturing of free DNA from contaminated samples using oligonucleotides coupled with paramagentic beads allowed the reduction of the detection time to six hours with a lower limit of 104 cells/mL.Da bi se odredio uzrok nestanka vakuuma u limenkama konzervirane hrane, obligatni anaerobi, termofilni organizmi koji stvaraju spore, izolirani su iz hrane u limenkama s glavnim sastojcima: povrće, rezanci i krumpir. Izolirane su termofilne bakterije iz 44 uzorka limenki uskladištenih pod anaerobnim uvjetima pri 37 °C barem 7 dana. Osim toga, ispitana su organska gnojiva upotrijebljena za uzgoj navedenog povrća pa su i iz tog izvora izolirane anaerobne termofilne bakterije. Identifikacija bakterijskih sojeva provedena je djelomičnim i potpunim sekvencioniranjem 16S-rRNA gena. Neke od dobivenih genskih sekvencija pokazale su visoki stupanj sličnosti s postojećim sekvencijama 16S-rRNA gena sojeva rodova Thermoanaerobacter, Thermoanaerobium i Thermoanaerobacterium. Do sada još nije bila ustanovljena važnost tih sojeva kao onečišćavača hrane. Tijekom identifikacije navedenih termofilnih bakterija autori su razvili genetički specifičan pristup utemeljen na PCR za određivanje izolata koji pripadaju rodovima Thermoanaerobacterium i Thermoanaerobacter. Izravno vezanje slobodne DNA iz onečišćenih uzoraka, koristeći oligonukleotide povezane s paramagnetskim zrncima omogućilo je smanjenje vremena detekcije na 6 sati s donjom granicom od 104 stanica/mL

First comparison of retroperitoneal versus transperitoneal robot-assisted nephroureterectomy with bladder cuff : a single center study

Introduction. After recent presentation of the first complete robot-assisted retroperitoneal nephroureterectomy with bladder cuff (RRNU) for patients with upper tract urothelial cancer (UTUC), we aimed to compare this new surgical technique with robot-assisted transperitoneal nephroureter ectomy (TRNU) representing the current standard of care. Methods. Robot-assisted nephroureterectomies (NUs) were retrospectively analyzed and compared based on two groups: transperitoneal versus retroperitoneal approach. Baseline data were collected for patient demographics, tumor characteristics, intra- (EAUiaiC) and postoperative (Clavien-Dindo) complications, and perioperative variables. Tumor characteristics included grade of malignancy, clinical stage, and surgical margin status. Short-term follow-up data including 30-day readmission rates were collected. Statistical analyses were performed assuming a significant p-value of < 0.05. Results. The analysis includes perioperative patient data after proven UTUC of 24 TRNU versus 12 RRNU (mean age: 70 versus 71 years; BMI: 25.9 versus 26.1 kg/m 2 ; CCI score ≥ 4: 83% versus 75%; ASA score ≥ 3: 37% vs 33%). Intraoperative (16.4% vs 0%, p = 0.35) and postoperative (25% vs 12.5%, p = 0.64) complications demonstrated no significant discrepancy. Notably, RRNU demonstrated significantly shorter surgery time (p < 0.05) and length of stay (p < 0.05). There was no significant difference in histopathological tumor characteristics, whereas significantly more lymph nodes were removed through RRNU (11.0±3.3 vs. 6.4±5.1, p < 0.05). Finally, no statistical difference was shown in short-term follow-up. Conclusion. We report the first head-to-head comparison between RRNU and TRNU. RRNU proves to be a safe and feasible approach which appears to be non-inferior to TRNU. RRNU expands the spectrum of minimally invasive treatment options, particularly for patients with major previous abdominal surgery

Intermatrix synthesis: easy technique permitting preparation of polymer-stabilized nanoparticles with desired composition and structure

The synthesis of polymer-stabilized nanoparticles (PSNPs) can be successfully carried out using intermatrix synthesis (IMS) technique, which consists in sequential loading of the functional groups of a polymer with the desired metal ions followed by nanoparticles (NPs) formation stage. After each metal-loading-NPs-formation cycle, the functional groups of the polymer appear to be regenerated. This allows for repeating the cycles to increase the NPs content or to obtain NPs with different structures and compositions (e.g. core-shell or core-sandwich). This article reports the results on the further development of the IMS technique. The formation of NPs has been shown to proceed by not only the metal reduction reaction (e.g. Cu0-NPs) but also by the precipitation reaction resulting in the IMS of PSNPs of metal salts (e.g. CuS-NPs)

Axonal mapping of the motor cranial nerves

Basic behaviors, such as swallowing, speech, and emotional expressions are the result of a highly coordinated interplay between multiple muscles of the head. Control mechanisms of such highly tuned movements remain poorly understood. Here, we investigated the neural components responsible for motor control of the facial, masticatory, and tongue muscles in humans using specific molecular markers (ChAT, MBP, NF, TH). Our findings showed that a higher number of motor axonal population is responsible for facial expressions and tongue movements, compared to muscles in the upper extremity. Sensory axons appear to be responsible for neural feedback from cutaneous mechanoreceptors to control the movement of facial muscles and the tongue. The newly discovered sympathetic axonal population in the facial nerve is hypothesized to be responsible for involuntary control of the muscle tone. These findings shed light on the pivotal role of high efferent input and rich somatosensory feedback in neuromuscular control of finely adjusted cranial systems

Value of c-MET and Associated Signaling Elements for Predicting Outcomes and Targeted Therapy in Penile Cancer

Whereas the lack of biomarkers in penile cancer (PeCa) impedes the development of efficacious treatment protocols, preliminary evidence suggests that c-MET and associated signaling elements may be dysregulated in this disorder. In the following study, we investigated whether c-MET and associated key molecular elements may have prognostic and therapeutic utility in PeCa. Formalin-fixed, paraffin-embedded tumor tissue from therapy-naïve patients with invasive PeCa was used for tissue microarray (TMA) analysis. Immunohistochemical staining was performed to determine the expression of the proteins c-MET, PPARg, β-catenin, snail, survivin, and n-MYC. In total, 94 PeCa patients with available tumor tissue were included. The median age was 64.9 years. High-grade tumors were present in 23.4%, and high-risk HPV was detected in 25.5%. The median follow-up was 32.5 months. High expression of snail was associated with HPV-positive tumors. Expression of β-catenin was inversely associated with grading. In both univariate COX regression analysis and the log-rank test, an increased expression of PPARg and c-MET was predictive of inferior disease-specific survival (DSS). Moreover, in multivariate analysis, a higher expression of c-MET was independently associated with worse DSS. Blocking c-MET with cabozantinib and tivantinib induced a significant decrease in viability in the primary PeCa cell line UKF-PeC3 isolated from the tumor tissue as well as in cisplatin- and osimertinib-resistant sublines. Strikingly, a higher sensitivity to tivantinib could be detected in the latter, pointing to the promising option of utilizing this agent in the second-line treatment setting

Evolutionary origins of hepatitis A virus in small mammals

The origins of human hepatitis A virus (HAV) are unknown. We conducted a targeted search for HAV-related viruses in small mammals sampled globally and discovered highly diversified viruses in bats, rodents, hedgehogs, and shrews. We demonstrate that these viruses share unique biological features with HAV, including structural, genomic, antigenic, and pathogenic properties. We found evidence of major shifts of HAV-related viruses between mammalian hosts in the past, suggesting both an origin of this viral genus in small mammals and a zoonotic origin of human HAV. Our data show that risk assessments for emerging viruses can benefit greatly from the analysis of viral infection patterns that evolved within animal reservoirs

Evolutionary origins of hepatitis A virus in small mammals

Hepatitis A virus (HAV) is an ancient and ubiquitous human pathogen recovered previously only from primates. The sole species of the genus Hepatovirus, existing in both enveloped and nonenveloped forms, and with a capsid structure intermediate between that of insect viruses and mammalian picornaviruses, HAV is enigmatic in its origins. We conducted a targeted search for hepatoviruses in 15,987 specimens collected from 209 small mammal species globally and discovered highly diversified viruses in bats, rodents, hedgehogs, and shrews, which by pairwise sequence distance comprise 13 novel Hepatovirus species. Near-complete genomes from nine of these species show conservation of unique hepatovirus features, including predicted internal ribosome entry site structure, a truncated VP4 capsid protein lacking N-terminal myristoylation, a carboxyl-terminal pX extension of VP1, VP2 late domains involved in membrane envelopment, and a cis-acting replication element within the 3Dpol sequence. Antibodies in some bat sera immunoprecipitated and neutralized human HAV, suggesting conservation of critical antigenic determinants. Limited phylogenetic cosegregation among hepatoviruses and their hosts and recombination patterns are indicative of major hepatovirus host shifts in the past. Ancestral state reconstructions suggest a Hepatovirus origin in small insectivorous mammals and a rodent origin of human HAV. Patterns of infection in small mammals mimicked those of human HAV in hepatotropism, fecal shedding, acute nature, and extinction of the virus in a closed host population. The evolutionary conservation of hepatovirus structure and pathogenesis provide novel insight into the origins of HAV and highlight the utility of analyzing animal reservoirs for risk assessment of emerging viruses

Epitope Structure of the Carbohydrate Recognition Domain of Asialoglycoprotein Receptor to a Monoclonal Antibody Revealed by High-Resolution Proteolytic Excision Mass Spectrometry

Recent studies suggest that the H1 subunit of the carbohydrate recognition domain (H1CRD) of the asialoglycoprotein receptor is used as entry site into hepatocytes by hepatitis A and B virus, and Marburg virus. Thus, molecules binding specifically to the CRD might exert inhibition towards these diseases by blocking the virus entry site. We report here the identification of the epitope structure of H1CRD to a monoclonal antibody by proteolytic epitope excision of the immune complex and high resolution MALDI-FTICR mass spectrometry. As a prerequisite of the epitope determination, the primary structure of the H1CRD antigen was characterised by ESI-FTICR-MS of the intact protein and by LCMS/MS of tryptic digest mixtures. Molecular mass determination and proteolytic fragments provided the identification of 2 intra-molecular disulfide bridges (7 Cys residues), and a Cysmercaptoethanol adduct formed by treatment with ß-mercaptoethanol during protein extraction. The H1CRD antigen binds to the monoclonal antibody in both native and Cysalkylated form. For identification of the epitope, the antibody was immobilized on N-hydroxysuccinimide activated Sepharose. Epitope- excision and - extraction with trypsin and FTICR-MS of affinity-bound peptides provided the identification of two specific epitope peptides, (5-16) and (17-23) which showed high affinity to the antibody. Affinity studies of the synthetic epitope peptides revealed independent binding of each peptide to the antibody

Isolierung und Expression des Leserasters fuer das VP1 des Hepatitis A virus fuer funktionelle und immunologische Untersuchungen

SIGLEAvailable from TIB Hannover: DW 6758 / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekDEGerman