The peptide trefoil factor family 3 (TFF3) is a major constituent of the intestinal mucus, playing an important role in the repair of epithelial surfaces. To further understand the role of TFF3 in the protection of intestinal epithelium, we tested the influence of TFF3 in a murine Toxoplasma gondii-induced ileitis model. Surprisingly, TFF3KO mice showed a reduced immune response in the ileum when compared to wild-type animals. Interleukin-12 and interferon-γ expression levels as well as the number of CD4+ lymphocytes were reduced in the infected TFF3KO mice. These effects were in line with the trend of elevated parasite levels in the ileum. Moreover, TFF1 expression was upregulated in the spleen of infected mice. These initial results indicate that TFF3 is involved in the immune pathology of T. gondii infection-induced intestinal inflammation. Thus far, the mechanisms of how TFF3 influences the immune response are not fully understood. Further studies should identify if TFF3 affects mucus sensing of dendritic cells and how TFF3 is involved in regulating the immune response as an intrinsic secretory peptide of immune cells.
A method for determining underivatized cholesterol and its oxygen derivatives (ChlOs) in animal tissue specimens and some food has been developed using capillary gas chromatography (GC) and mass spectrometry (MS) with 5α-cholestane as the internal standard. The analysis preparation procedure for cholesterol and ChlOs consisted of lipid extraction followed by gentle saponification using 2 M KOH in ethanol. The analyzed samples were saponified overnight at room temperature in the dark with shaking. Then, free cholesterol and ChlOs were extracted using diethyl ether. The extracts were dried under argon, and the residues were redissolved in chloroform before GC-MS analysis. Cholesterol was analyzed in biological materials using the short column temperature program. The simultaneous quantification of cholesterol and its oxides in biological materials was performed using the longer column temperature program. Cholesterol was eluted faster than ChlOs. The chromatographic method that used column temperature program A can be successfully used for the simple and rapid quantification of cholesterol in biological materials, while the method using column temperature program B is more suitable for the simultaneous quantification of cholesterol and its oxides in biological materials.
Influenza A virus (IAV) infection causes an acute respiratory disease characterized by a strong inflammatory immune response and severe immunopathology. Proinflammatory mechanisms are well described in the murine IAV infection model, but less is known about the mechanisms leading to the resolution of inflammation. Here, we analyzed the contribution of CD11b+Ly6C++Ly6G− cells to this process. An accumulation of CD11b+Ly6C++Ly6G− cells within the lungs was observed during the course of IAV infection. Phenotypic characterization of these CD11b+Ly6C++Ly6G− cells by flow cytometry and RNA-Seq revealed an activated phenotype showing both pro- and anti-inflammatory features, including the expression of inducible nitric oxide synthase (iNOS) by a fraction of cells in an IFN-γ-dependent manner. Moreover, CD11b+Ly6C++Ly6G− cells isolated from lungs of IAV-infected animals displayed suppressive activity when tested in vitro, and iNOS inhibitors could abrogate this suppressive activity. Collectively, our data suggest that during IAV infection, CD11b+Ly6C++Ly6G− cells acquire immunoregulatory function, which might contribute to the prevention of pathology during this life-threatening disease.
This is the first report of Pasteurella multocida type B in Hungarian pigs. This disease was observed in backyard-raised pigs in three households within a small area. Neither the source of the infection nor the epidemiological connection between any of the premises could be determined. The most consistent lesion was dark red discolouration of the skin of the ventral neck and brisket, with accompanying oedema and haemorrhages. The morbidity was low and lethality relatively high, with three dead (50%) and two euthanised (33%) out of six affected animals. A total of three isolates of P. multocida (P55, P56 and P57) were cultured from these cases and examined in detail. These were identified as P. multocida ssp. multocida biovar 3. All were toxA negative and belonged to serotype B:2. Multilocus sequence typing was used to assign these to a new sequence type (ST61) that is closely related to other haemorrhagic septicaemia causing strains of P. multocida regardless of the host. M13 polymerase chain reaction and virulence-associated gene typing also show that type B strains form a highly homogeneous, distinct phylogenic group within P. multocida.
The lactic acid bacteria are key microorganisms for the production and preservation of fermented dairy products, cheeses, sourdough bread, and lacto-fermented vegetables. This study was developed to monitor lactic acid produced by Lactobacillus plantarum ATCC 8014 and Lactobacillus casei ATCC 393, as single strains and combined, in fermenting media by Fourier Transform Infrared Spectroscopy coupled to multivariate statistical analysis. Media containing different mixtures of carbohydrates were chosen as model fermenting media for monitoring lactic acid concentration by infrared spectroscopy, due to the fact that vegetable and animal food matrices could contain different carbohydrates as carbon sources. Three different types of media were obtained by adding different carbohydrates to a basic MRS medium. HPLC was used as reference method for lactic acid quantification. The calibration set (n=36) was used for building model, while a validation set (n=13) for testing the robustness of the developed model. The coefficients of determination between predicted and reference values were 0.986 and 0.965, while root mean square error for calibration and validation sets recorded values of 0.127 and 0.263 g·l−1, respectively. Results confirmed the efficiency of FTIR spectroscopy combined with multivariate statistics, as a rapid, reliable, and cost-effective tool for routine monitoring of lactic acid.
Morbidity and mortality rates during the COVID-19 pandemic have been particularly high among elderly people (>65 years). This review summarises some of the important physiological and clinical aspects in the background of augmented risk. Airway clearance provides defence against inhaled particles, including viruses. Some relevant studies have indicated that clearance from the small and large airways is slower in elderly people. Cough peak flow (the speed of expiratory airflow during coughing, or cough power) is another important parameter that reflects the defence capacity of the respiratory system. Age has likewise been shown to induce inspiratory and expiratory muscle weakness and, as a consequence, a low cough peak flow. In addition to the weakening of these non-specific defences in elderly people, the specific immune response against the SARS-CoV-2 virus has been found to be nearly blocked in aged mice, and the augmented synthesis of prostaglandin D2 (PGD2) was found to play a role in this phenomenon. Aged animals were protected from death by a specific antagonist of PGD2. Among aged people suffering from COVID-19, there were disproportionally more patients with low CD8 T lymphocyte counts and high plasma concentrations of interleukin 6 (IL-6). This combination of deficient cellular immunity and overt inflammatory response in COVID-19 has been identified as a significant risk factor of mortality.
of piglets [ 4 ]. Piglets exposed to this MRSA concentration for 24 h in an aerosol chamber were persistently colonized with MRSA. An airborne MRSA concentration of 10 4 cfu/m 3 resulted in transiently colonized animals. In contrast to the mean
Bevezetés: A sejten kívüli szabad DNS-t már az 1940-es években
kimutatták. Eredetéről több elmélet is létezik: lehetséges folyamat a tumoros
sejtekből, valamint ezzel párhuzamosan az egészséges sejtekből történő
felszabadulás is. Célkitűzés: Munkánk célja a szabad DNS
felszabadulási ütemének vizsgálata volt SHO-egér/HT-29 humán colorectalis
adenocarcinoma sejtvonal xenograftmodellben, valamint célul tűztük ki egészséges
és C38 tumorral oltott C57BL/6-os egerek véráramába juttatott mesterségesen
fölszaporított metilált és nem metilált DNS-szakaszok lebomlásának nyomon
követését. Módszer: SHO-egerekre HT-29 sejteket oltottunk
subcutan, majd vért vettünk 8 héten keresztül. A plazma
szeparálása után DNS-t izoláltunk, majd mitokondriális és genomiális
RT-PCR-próbákkal megállapítottuk a humán/egér DNS-arányt. A szabad DNS
lebomlásának vizsgálatához egészséges és C38 tumorsejttel oltott C57BL/6-os
állatok vérébe 3000 bázispár (bp) méretű in vitro metilált és
nem metilált DNS-fragmentumot juttattunk. Az amplikonok degradációját 19 valós
idejű PCR-próbával mértük, a bomlás ütemére a relatív amplikonkoncentrációk
alapján következtettünk. Eredmények: A tumorból származó humán
DNS mennyisége a 2. hétig a kimutathatósági határ alatt volt, majd a 3. héttől
folyamatos emelkedést tapasztaltunk, amely a 8. hétre 18,26%-ot ért el. A
véráramba juttatott DNS-szakaszok lebomlásának sebességében különbséget
mutattunk ki a nem metilált és a metilált fragmentumok között. Az egészséges
állatokban a nem metilált DNS 6 óra után eltűnt a vérplazmából, míg a metilált
fragmentum szakaszai 24 óra múlva is kimutathatók voltak. Tumoros állatokban a
degradáció mértéke lelassult, és mindkét forma kimutathatóvá vált 24 óra
elteltével. Következtetés: A szabad DNS szerepének és
hatásmechanizmusának vizsgálatát egyre nagyobb érdeklődés övezi. Munkánk
segítséget nyújthat a DNS felszabadulásának és degradációjának pontosabb
megismeréséhez. Orv Hetil. 2018; 159(6): 223–233.